OA20663A - 2-formyl-3-hydroxyphenyloxymethyl compounds capable of modulating hemoglobin. - Google Patents

Abstract

The present disclosure relates generally to compounds and pharmaceutical compositions suitable as modulators of hemoglobin, and methods for their use in treating disorders mediated by hemoglobin.

Description

MODULATORS OF HEMOGLOBIN

CROSS-REFERENCE TO RELATE!) APPLICATIONS

[0001] Thîs application daims the benefit under 35 U.S.C. §119(e) of United States Provisional Application No. 62/769,196, filed November 19, 2018, United States Provisional Application No. 62/821,314, filed Match 20, 2019, United States Provisional Application No. 62/848,773, filed May 16, 2019, and United States Provisional Application No. 62/883,313, filed August 6, 2019, each of whîch is hereby incorporaled by reference in its entirety.

FIELD

[0002] Provided herein are compounds and pharmaceutical compositions suitable as modulators of hemoglobin, and methods for their use in treating disorders mediated by hemoglobin.

BACKGROUND

[00031 Sickle cell dîsease is a disorder of the red blood cells, found particularly among those of African and Mediterranean descent. The basis for sickle cell disease is found in sickle hemoglobin (HbS), which contains a point mutation relative to the prévalent peptide sequence of hemoglobin A (HbA).

[0004] Hemoglobin (Hb) transports oxygen molécules 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 where glutamic acid is replaced with valine, making HbS susceptible to polymerization under hypoxie conditions to gîve the HbS containing red blood cells their characteristic sickle shape. The sickled cells are also more rigid than normal red blood cells, and their lack of flexibility can lead to blockage of blood vessels.

[0005] 24iydroxy-6-((2-(l-isopropyl-lH-pyrazol-5-yl)pyridin-3-yl)methoxy)benzaldehyde (also known as voxelotor or GBT440), a modulator of hemoglobin that increases the affinity of hemoglobin for oxygen and consequently inhibîts polymerization of HbS when subjected to hypoxie conditions, is currently in Phase 3 clinîcal trials for the treatment of sickle cell disease (NCT03036813).

[0006] WO 2014/150268 describes modulators of hemoglobin that are structurally related to the compounds disclosed herein.

[0007] A need exists for compounds that can treat disorders that are mediated by abnormal Hb such as HbS and methods of treating such disorders. Compounds that hâve an improved pharmacokinetic profile relative to known modulators of hemoglobin while maintaining or improvîng efficacy are of particular interest, as such compounds would allow for favorable dosing regtmens (e.g., lower and/or less frequent doses).

SUMMARY

[0008]

Provïded herein is a compound of formula Ii

Z

H

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

X is CH or N;

Y is CH or N;

Z is absent, CH2, O, or S; and

R¹ is mono-hydroxy-(Ci-4alkyI), di-hydroxy-(Ci-4 alkyl), -CH2CH2OCH3, -CH2CH2CN, or

[0009] Some embodiments provide for pharmaceutical compositions comprising a compound as described herein, or an isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable sait of each thereof, and a pharmaceutically acceptable excipient. Some embodiments provide for pharmaceutical compositions comprising a compound as described herein or a pharmaceutically acceptable sait thereof, and a pharmaceutically acceptable excipient. Some embodiments provide for pharmaceutical compositions comprising a compound as described herein and a pharmaceutically acceptable excipient.

[0010] Also provided hereln are methods for increasing oxygen affinity of hemoglobîn (e.g., hemoglobin S) in a subject in need thereof, comprising administering to the subject a compound as described herein or a pharmaceutîcal composition as described herein.

[0011] Also provided herein are methods for treating a disorder mediated by hemoglobin in a subject in need thereof, comprising administering to the subject a compound as described herein or a pharmaceutîcal composition as described herein.

[0012] Also provided herein are methods for treating sickle cell disease in a subject in need thereof, comprising administering to the subject a compound as described herein or a pharmaceutîcal composition as described herein.

DETAILED DESCRIPTION

Définitions

[0013] As used in the présent spécification, the following words, phrases and symbols are generally intended to hâve the meanings as set forth below, except to the extent that the context in which they are used indicates otherwise.

[0014] A dash that is not between two letters or symbols is used to indicate a point of attachment for a substituent. For example, -C(O)NHi is attached through the carbon atom. A dash at the front or end of a Chemical group is a matter of convenience; Chemical groups may be depicted with or without one or more dashes without losing their ordinary meaning. A wavy line or a dashed line drawn through or perpendicular across the end of a line in a structure indicates a specified point of attachment of a group. Unless chemically or structurally required, no directionality or stereochemistry is îndicated or împlîed by the order in which a Chemical group is written or named.

[0015] The prefix “C_u-v” indicates that the following group has from u to v carbon atoms. For example, “Ci-6 alkyl” indicates that the alkyl group has from 1 to 6 carbon atoms. In another example, “Ci-4 alkyl” indicates that the alkyl group has from 1 to 4 carbon atoms.

[0016] Reference to “about” a value or parameter herein includes (and describes) embodiments that are directed to that value or parameter per se. In certain embodiments, the term “about” includes the îndicated amount ± 10%. In other embodiments, the term “about” includes the indicated amount ± 5%. In certain other embodiments, the term “about” includes the îndicated amount ± 1%. Also, to the term “about x” includes description of “x”. Also, the singular forms “a” and “the” include plural référencés unless the context clearly dictâtes otherwise. Thus, e.g., référencé to ‘the compound includes a plurality of such compounds and référencé to “the assay” includes référencé to one or more assays and équivalents thereof known to those ski lied in the art.

[0017] “AlkyI” refers to an unbranched or branched saturated hydrocarbon chain. As used herein, alkyl has 1 to 20 carbon atoms (Le., Ci.20 alkyl), 1 to 12 carbon atoms (Le., Ci-12 alkyl), 1 to 8 carbon atoms (Le., Ci-s alkyl), 1 to 6 carbon atoms (Le., Ci-e alkyl) or 1 to 4 carbon atoms (Le., Cm alkyl). Examples of alkyl groups include, e.g., methyl, ethyl, propyl, isopropyl, nbutyl, sec-butyl, iso-butyi, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3hexyl and 3-methylpentyl. When an alkyl residue having a spécifie number of carbons is named by Chemical name or identified by molecular formula, ali positional isomers having that number of carbons may be encompassed; thus, for example, “butyl” includes n-butyl (Le., -(CH2)₃CH₃), sec-butyl (Le., -CH(CH₃)CH2CH₃), isobutyl (Le., -CH₂CH(CH₃)₂) and tert-butyl (Le., -C(CH₃)₃); and “propyl” includes n-propyl (Le., -(CH2)2CH₃) and isopropyl (Le., -CH(CH₃)₂).

[0018] Certain 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 referred to as an “alkylene” group or an “alkylenyl” group, an “arylene” group or an “arylenyl” group, respectively. Also, unless indicated explicitly otherwise, where combinations of groups are referred to herein as one moiety, e.g., arylalkyi or aralky 1, the last-mentioned group contains the atom by which the moiety is attached to the rest of the molécule.

[0019] “Alkenyl” refers to an alkyl group contai ning at least one carbon-carbon double bond and having from 2 to 20 carbon atoms (Le., C2-20 alkenyl), 2 to 8 carbon atoms (Le., C2-8 alkenyl), 2 to 6 carbon atoms (Le., C2-6 alkenyl) or 2 to 4 carbon atoms (Le., C2.4 alkenyl).

Examples of alkenyl groups include, e.g., ethenyl, propenyl, butadienyl (including 1,2butadienyl and 1,3-butadienyi).

[0020] “Aîkynyl” refers to an alkyl group containing at least one carbon-carbon triple bond and having from 2 to 20 carbon atoms (Le., C2-20 alkynyl), 2 to 8 carbon atoms (Le., C2-8 alkynyl), 2 to 6 carbon atoms (Le., Cz^ alkynyl) or 2 to 4 carbon atoms (Le., C2-4 alkynyl). The term “alkynyl” also includes those groups having one triple bond and one double bond.

[0021] “Alkoxy” refers to the group “alkyl-O-”. Examples of alkoxy groups include, e.g., methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, nhexoxy and 1,2-dimethylbutoxy.

[0022] “Alkylthio” refers to the group “alkyl-S-”. “AlkyIsulfinyl” refers to the group “alkylS(O)-”. “Alkylsulfonyl” refers to the group “alkyl-S(O)2-”.

[0023] “Acyl” refers to a group -C(O)R^y, wherein R^y is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyciyl, aryl, heteroalkyi or heteroaryl; each of which may be optionally 5 substituted, as defined herein. Exampies of acyl include, e.g., formyi, acetyl, cyclohexylcarbonyl, cyclohexyimethyl-carbonyl and benzoyl.

[0024] “Amido” refers to both a “C-amido” group which refers to the group -C(O)NR^yR^z and an “N-amido” group which refers to the group -NR^yC(O)R^z, wherein R^y and R² are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyciyl, aryl, heteroalkyi or 10 heteroaryl; each of which may be optionally substituted, as defined herein, or R^y and R^z are taken together to form a cycloalkyl or heterocyciyl; each of which may be optionally substituted, as defined herein.

[0025] “Amino” refers to the group -NR^yR^z wherein R^y and R^z are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyciyl, aryl, heteroalkyi or heteroaryl; each of which 15 may be optionally substituted, as defined herein.

[0026] “Amîdino” refers to -C(NR^y)(NR^z2), wherein R^y and R^z are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyciyl, aryl, heteroalkyi or heteroaryl; each of which may be optionally substituted, as defined herein.

[0027] “Aryl” refers to an aromatic carbocyclîc group having a single ring (e.g., monocyclic) 20 or multiple rings (e.g., bicyclic or tricyclic) including fused Systems. As used herein, aryl has 6 to 20 ring carbon atoms (Le., Cô-zo aryl), 6 to 12 carbon ring atoms (Le., CA 12 aryl), or 6 to 10 carbon ring atoms (Le., Cmo aryl). Examples of aryl groups include, e.g., phenyl, naphthyl, fluorenyl and anthryl. Aryl, however, does not encompass or overlap in any way with heteroaryl defined below. If one or more aryl groups are fused with a heteroaryl, the resulting ring system 25 is heteroaryl regardless of the point of attachment. If one or more aryl groups are fused with a heterocyciyl, the resulting ring system is heterocyciyl regardless of the point of attachment.

[0028] “Arylalkyl” or “Araikyl” refers to the group “aryl-alkyl-”.

[0029] “Carbamoyl” refers to both an “O-carbamoyl” group which refers to the group -O-C(O)NR^yR^z and an “N-carbamoyl” group which refers to the group -NR^yC(O)OR^z, wherein 30 R^y and R^z are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyciyl, aryl, heteroalkyi or heteroaryl; each of which may be optionally substituted, as defined herein.

) [0030] “Carboxyl ester” or “ester” refer to both -0C(0)R^x and -C(O)OR^X, whereîn R^x is alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted, as defined herein.

[0031] “Cycloalkyl” refers to a saturated or partially unsaturated cyclic alkyl group having a single ring or multiple rings including fused, bridged and spiro ring Systems. The term “cycloalkyl” includes cycloalkenyl groups (Le., the cyclic group having at least one double bond) and carbocyclic fused ring Systems having at least one sp³ carbon alom (û., at least one non-aromatic ring). As used herein, cycloalkyl has from 3 to 20 ring carbon atoms (Le., C3-20 cycloalkyl), 3 to 12 ring carbon atoms (Le., C3-12 cycloalkyl), 3 to 10 ring carbon atoms (Le., C?.

jo cycloalkyl), 3 to 8 ring carbon atoms (Le., C3-8 cycloalkyl), or 3 to 6 ring carbon atoms (Le., C3-6 cycloalkyl). Monocyclic groups include, for example, cyclopropyl, cyclobutyl, cyclopenlyl, cyclohexyl, cycloheptyl and cyclooctyl. Poiycyclic groups include, for example, bicyclo[2.2.1]heptanyl, bicyclo[2,2.2]octanyl, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl and the like. Furlher, the term cycloalkyl is întended to encompass any non-aromatic ring which may be fused to an aryl ring, regardless of the attachment to the remainder of the molécule. Still further, cycloalkyl also includes “spirocycloalkyl” when there are two positions for substitution on the same carbon atom, for example spiro [2.5 ]octanyl, spîro[4.5]decanyl, or spiro[5.5]undecanyl.

[0032] “Cycloalkylalkyl” refers to the group “cycloalkyl-alkyl-”.

[0033] “Guanidino” refers to -NR^yC(=NR^z)(NR^yR^z), wherein each 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.

[0034] “Imino” refers to a group -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 25 of which may be optionally substituted, as defined herein.

[0035] “Imido” refers to a group -C(O)NR^yC(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.

[0036] “Halogen” or “halo” refers to atoms occupying group VIIA of the periodic table, such 30 as fluoro, chloro, bromo or iodo.

P [0037] “Haloalkyl” refers to an unbranched or branched alky 1 group as defïned above, wherein one or more (e.g., 1 to 6, or 1 to 3) hydrogen atoms are replaced by a halogen. For example, ' where a residue is substituted with more than one haiogen, it may be referred to by using a prefix corresponding to the number of halogen moieties attached. Dihaloalkyl and trihaloalkyl refer to alkyI substituted with two (“di”) or three (“tri”) halo groups, which may be, but are not necessarily, the same halogen. Examples of haloalkyl include, e.g., trifluoromethyl, difluoromethy], fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl and the like.

[0038] “Haloalkoxy” refers to an alkoxy group as defined above, wherein one or more (e.g., 1 10 to 6, or 1 to 3) hydrogen atoms are replaced by a halogen.

[0039] “Hydroxyalkyl” refers to an alkyl group as defined above, wherein one or more (e.g., 1 to 6, or 1 to 3) hydrogen atoms are replaced by a hydroxy group. A “mono-hydroxy-(Ci-4 alkyl)” refers to a Ci-4 alkyl group as defined above, wherein one hydrogen atom is replaced by a hydroxy group. A “di-hydroxy-(Cî-4 alkyl)” refers to a Cm alkyl group as defined above, wherein two hydrogen atoms are replaced by hydroxy groups.

[0040] “Heteroalkyl” refers to an alkyl group in which one or more of the carbon atoms (and any assocîated hydrogen atoms) are each independently replaced with the same or different heteroatomic group, provided the point of attachment to the remainder of the molécule is through a carbon atom. The term “heteroalkyl” includes unbranched or branched saturated chain having carbon and heteroatoms. By way of example, 1, 2 or 3 carbon atoms may be independently replaced with the same or different heteroatomic group. Heteroatomic groups include, but are not limited to, -NR^y-, -O-, -S-, -S(O)-, -S(O)2-, and the like, wherein 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 heteroalkyl groups include, e.g., ethers (e.g., -CH₂OCH₃, -CH(CH₃)OCH₃, -CH₂CH₂OCH₃, CH CH-OOITHOCfl·. etc.), thioethers (e.g., -CH₂SOF, -CH(CH₃)SCH₃, -CH₂CH₂SCH₃, CH2CH2SCH2CH2SCH3, etc.), sulfones (e.g., -CH₂S(O)₂CH₃, -CH(CH₃)S(O)₂CH₃, CH₂CH₂S(O)2CH₃, -CH₂CH₂S(O)₂CH₂CH₂OCH₃, etc.) and amines (e.g., -CH₂NR^yCH3, CH(CH3)NR^yCH3, -CH₂CH₂NR^yCH₃,

-CH₂CH₂NR^yCH2CH2NR^yCH3, etc., where 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.

P [0041] “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 includes 1 to 20 ring carbon atoms (Le., C1-20 heteroaryl), 3 to 12 ring carbon atoms (Le., C3.12 heteroaryl), or 3 to 8 carbon ring atoms (Le., C3-8 heteroaryl), and 1 to 5 ring heteroatoms, 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom independently selected from nitrogen, oxygen and sulfur. In certain instances, heteroaryl includes 5-10 membered ring Systems, 5-7 membered ring Systems, or 5-6 membered ring Systems, each independently having 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom independently selected from

1Ü nitrogen, oxygen and sulfur. Examples of heteroaryl groups include, e.g., acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzofuranyl, benzothiazolyl, benzothiadiazolyl, benzonaphthofuranyl, benzoxazolyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[l,2-a]pyridyl, carbazolyl, cinnolinyl, dibenzofuranyl, dîbenzothiophenyl, furanyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, isoquinolyl, isoxazolyl, naphthyridinyl, oxadiazolyl, oxazolyl, 1-oxidopyridinyl, 1-oxidopyrimîdinyl, 1oxidopyrazinyl, 1-oxidopyridazinyl, phenazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrîdinyl, pyrazinyl, pyrimîdinyl, pyrîdazînyl, quînazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, thiazolyl, thiadiazolyl, triazoiyl, tetrazolyl and triazînyl. Examples of the fused-heteroaryl rings include, but are not limited to, benzo[d]thîazolyi, quinolinyl, isoquinolinyl, benzo[b]thiophenyl, indazolyl, benzo[d]imidazolyl, pyrazolo[l,5-a]pyridinyl and imidazo[l,5-a]pyridinyl, where the heteroaryl can be bound via either ring of the fused System. Any aromatic ring, having a single or multiple fused rings, containing at least one heteroatom, is considered a heteroaryl regardless of the attachaient to the remainder of the molécule (Le., through any one of the fused rings). Heteroaryl does not encompass or overlap with aryl as defined above.

[0042] “Heteroarylalkyl” refers to the group “heteroaryl-alkyl-”.

[0043] “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 (Le., the heterocyclyl group having at least 30 one double bond), brïdged-heterocyclyl groups, fused-heterocyclyl groups and spiroheterocyclyl groups. A heterocyclyl may be a single ring or multiple rings wherein the multiple rings may be fused, bridged or spiro, and may comprise one or more (e.g., 1 to 3) oxo (=0) or N-oxide (-Θ') moîeties. Any non-aromatic ring containing at least one heteroatom is considered a heterocyclyl, regardless of the attachment (Le., can be bound through a carbon atom or a heteroatom). Further, the tenu heterocyciyl is intended to encompass any non-aromatic ring containing at ieast one heteroatom, which ring may be fused to an aryl or heteroaryl ring, regardless of the attachaient to the remainder of the molécule. As used herein, heterocyciyl has 2 to 20 ring carbon atoms (i.e., C2-20 heterocyciyl), 2 to 12 ring carbon atoms (i.e., C2-12 heterocyciyl), 2 to 10 ring carbon atoms (i.e., C2.10 heterocyciyl), 2 to 8 ring carbon atoms (i.e., C2-8 heterocyciyl), 3 to 12 ring carbon atoms (i.e., C3-12 heterocyciyl), 3 to 8 ring carbon atoms (i.e., C3-8 heterocyciyl), or 3 to 6 ring carbon atoms (i.e., C3-6 heterocyciyl); having 1 to 5 ring heteroatoms, 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom independently seiected from nitrogen, sulfur or oxygen. Examples of heterocyciyl groups include, e.g., azetidinyl, azepinyl, benzodioxolyl, benzo[b][l,4]dioxepinyl, 1,4benzodîoxanyi, benzopyranyl, benzodioxinyl, benzopyranonyl, benzofuranonyl, dioxolanyl, dîhydropyranyl, hydropyranyl, thienyl[l,3]dithianyl, decahydroisoquînolyl, furanonyl, imidazolinyi, imîdazolidinyl, indolinyl, indolizinyl, isoindolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoîndolyl, 2-oxopiperazînyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, oxiranyl, oxetanyl, phenothiazinyl, phenoxazinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolîdinyl, tetrahydrofury], tetrahydropyranyl, trithîanyl, tetrahydroquinolinyl, thiophenyl (i.e., thienyl), tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl and 1,1-dioxo-thiomorpholinyl. The terni “heterocyciyl” also includes “spîroheterocyclyl” when there are two positions for substitution 011 the sanie carbon atom. Examples of the spiroheterocyclyl rings include, e.g., bicyclic and tricyclîc ring Systems, such as 2-oxa-7azaspiro[3.5]nonanyl, 2-oxa-6-azaspîro[3.4]octanyl and 6-oxa-l-azaspiro[3.3]heptanyl.

Examples of the fused-heterocyclyl rings include, but are not limited to, 1,2,3,4tetrahydroisoquinolinyl, 4,5,6,7-tetrahydrothieno[2,3-c]pyridinyl, indolinyl and isoindolinyl, where the heterocyciyl can be bound via either ring of the fused System.

[0044] “I-Ieterocyclylalkyl” refers to the group “heterocyciyl-alkyl-.”

[0045] “Oxime” refers to the group -CR^y(=NOH) wherein R^y is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyciyl, aryl, heteroalkyi or heteroaryl; each of which may be optionally substituted, as defined herein.

[0046] “Sulfonyl” refers to the group -S(O)2R^y, where R^y is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyciyl, aryl, heteroalkyi or heteroaryl; each of which may be optionally substituted, as defined herein. Examples of sulfonyl are methylsulfonyl, ethylsulfonyl, phenylsulfonyi and toluenesulfonyl.

[0047] “Sulfinyl” refers to the group -S(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 hereîn. Examples of sulfinyl are methylsulfinyl, ethylsulfinyl, phenylsulfinyl and toluenesulfinyl.

[0048] “Sulfonamido” refers to the groups -SO2NR^yR^z and -NR^ySO2R^z, where R^y and R^z are each îndependently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted, as defined hereîn.

[0049] The terms “optional” or “optionally” means that the subsequently described event or circumstance may or may not occur and that the description includes instances where said event or circumstance occurs and instances in which it does not. Aiso, the term “optionally substituted” refers to any one or more (e.g., 1 to 5, or 1 to 3) hydrogen atoms on the designated atom or group may or may not be replaced by a moiety other than hydrogen.

[0050] The term “substituted” used hereîn 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) hydrogen atom is replaced by a bond to a non-hydrogen atom such as, but not limited to alkyl, alkenyl, alkynyl, alkoxy, alkylthio, acyl, amido, amino, amidino, aryl, aralkyl, azido, carbamoyl, carboxyl, carboxyl ester, cyano, cycloalkyl, cycloalkylalkyl, guanidino, halo, haloalkyl, haloalkoxy, hydroxyalkyl, heteroalkyl, heteroaryl, heteroarylaikyl, heterocyclyl, heterocyclylalkyl,

- NHNH2, =NNH₂, imino, imido, hydroxy, oxo, oxime, nitro, sulfonyl, sulfinyl, alkylsulfonyl, aikylsulfinyl, thîocyanate, -S(O)OH, -S(O)₂OH, sulfonamido, thiol, thioxo, N-oxide or

- Si(R^y)3, wherein each R^y is îndependently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, aryl, heteroaryl or heterocyclyl.

[0051] in certain 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 îndependently replaced with deuterium, halo, cyano, nitro, azido, oxo, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl,

- NR^eR^h, -NR^ÊC(=O)R^h, -NR^êC(=O)NR^gR^h, -NR^C(=O)OR^h, -NRÂS(=O)i.2R^h, -C(=O)R*, -C(-O)OR^@, -OC(=O)OR^g, -OCOOiRA -C(=O)NR^sR^h, -OC(=O)NR$R^h, -OR^S, -SR^S, -S(=O)R», -S(=O)2R^g, -OS(=O)p2R^ë, -S(=O)i-2OR8, -NR®S(=O)i-2NRSR^h, =NSO2R^g, - NORA S(=O)i-2NR^eR^h, -SF5, -SCF3 or -OCF3. In certain 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 with

I 1 • -C(-O)R^g, -C(=0)0R^g, -C(=O)NR^gR^h, -CH2SO2R^Ë, or -CH2SO₂NR®R^h. In the foregoing, R^g and R^h are the sanie or different and independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, and/or heteroarylalkyl. In certain 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 a bond to an amino, cyano, hydroxyi, imino, nitro, oxo, thioxo, halo, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, and/or heteroarylalkyl, or two of R^g and R^h and R^! are taken together with the atoms to which they are attached to form a heterocyclyl ring optionally

1Ü substituted with oxo, halo or alkyl optionally substituted with oxo, halo, amino, hydroxyi, or alkoxy.

[0052] Polymers or similar indefinite structures arrived at by defining substituents with further substituents appended ad infinitum (e.g., a substituted aryl having a substituted alkyl which is itself substituted with a substituted aryl group, which is further substituted by a substituted heteroalkyl group, etc.) are not intended for inclusion herein. Unless otherwise noted, the maximum number of serial substitutions in compounds described herein is three. For example, serial substitutions of substituted aryl groups with two other substituted aryl groups are limited to ((substituted aryl)substituted aryl) substituted aryl. Similarly, the above définitions are not intended to include impermissible substitution patterns (e.g., methyl substituted with 5 fluorines or heteroaryl groups having two adjacent oxygen ring atoms). Such impermissible substitution patterns are well known to the skilled artisan. When used to modify a Chemical group, the term “substituted” may descrîbe other Chemical groups defined herein.

[0053] In certain embodiments, as used herein, the phrase “one or more” refers to one to five. In certain embodiments, as used herein, the phrase “one or more” refers to one to three.

[0054] Any compound or structure given herein, îs intended to represent unlabeled forms as well as isotopically labeled forms (isotopologues) of the compounds. These forms of compounds may also be referred to as and include “isotopically enriched analogs.” Isotopically labeled compounds hâve structures depicted herein, except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine and iodine, such as ²H, ³H, ^HC, ¹³C, ¹⁴C, ⁱ³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O,³¹P, ³²P, ³⁵S, ¹⁸F, ³⁶C1,¹²³I, and ¹²⁵I, respectively. Various isotopically labeled compounds of the présent disclosure, for example those into which radioactive isotopes such as ³H, ¹³C and ¹⁴C are încorporated. Such isotopically labelled compounds may be useful in metabolic studies, reaction kinetic studies, détection or imaging techniques, such as positron émission tomography (PET) or single-photon émission compuled tomography (SPECT) inciuding drug or substrate tissue distribution assays or in radioactive treatment of patients.

[0055] The term “isotopically enriched analogs” includes “deuterated analogs” of compounds described herein in which one or more hydrogens is/are replaced by deuterium, such as a hydrogen on a carbon alom. Such compounds exhibit increased résistance to metabolism and are thus useful for increasing the half-life of any compound when administered to a mammal, particularly 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 means well known in the art, for example by employing starting materials in which one or more hydrogens hâve been replaced by deuterium.

[0056] Deuterium labelled or substituted therapeutic compounds of the disclosure may hâve improved DMPK (drug metabolism and pharmacokinetics) properties, relating to distribution, metabolism and excrétion (ADME). Substitution with heavier isotopes such as deuterium may afford certain therapeutic ad vanta ges resulting from greater metabolic stabiiity, for example increased in vivo half-life, reduced dosage requirements and/or an împrovement în therapeutic index. An ¹⁸F, ³H, ^l,C labeled compound may be useful for PET or SPECT or other imaging studies. Isotopically labeled compounds of this disclosure and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the schemes or in the examples and préparations described below by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent. It is understood that deuterium in this context is regarded as a substituent in a compound described herein.

[0057] The concentration of such a heavier isotope, specifically deuterium, may be defîned by an isotopic enrichment factor. In the compounds of this disclosure any alom not specifically designated as a particular isotope is meant to represent any stable isotope of that atom. Unless otherwise stated, when a position is designated specifically as “H” or “hydrogen”, the position is understood to hâve hydrogen at its natural abundance isotopic composition, Accordingly, in the compounds of this disclosure any atom specifically designated as a deuterium (D) is meant to represent deuterium. Further, in some embodiments, the corresponding deuterated analog is provided.

[0058] In many cases, the compounds of this dîsclosure are capable of forming acid and/or base salts by vjrtue of the presence of amino and/or carboxyl groups or groups similar thereto.

[0059] Provided also are a pharmaceutically acceptable sait, isotopically enriched analog, deuterated analog, isomer (such as a stereoisomer), mixture of isomers (such as a mixture of stereoisomers), prodrug, and métabolite of the compounds described herein.

[0060] “Pharmaceutically acceptable” or “physiologically acceptable” refer to compounds, salts, compositions, dosage forms and other materials which are useful in preparing a pharmaceutical composition thaï is suitable for veterînary or human pharmaceutical use.

[0061] The term “pharmaceutically acceptable sait” of a given compound refers to salts that retain the biologîcal effectiveness and properties of the given compound and which are not biologically or otherwise undesîrable. “Pharmaceutically acceptable salts” or “physiologically acceptable salts” include, for example, salts with inorganic acids and salts with an organic acid. In addition, if the compounds described herein are obtained as an acid addition sait, the free base can be obtained by basifying a solution of the acid sait. Conversely, if the product is a free base, an addition sait, particularly a pharmaceutically acceptable addition sait, may be produced by dissolving the free base in a suitable organic solvent and treating the solution with an acid, in accordance with conventional procedures for preparing acid addition salts from base compounds. Those skilled in the art will recognize various synthetic méthodologies that may be used to préparé nontoxic pharmaceutically acceptable addition salts. Pharmaceutically acceptable acid addition salts may be prepared from inorganic and organic acids. Salts derived from inorganic acids include, e.g., hydrochloric acid, hydrobromic acid, sul furie acid, nitric acid, pho.sphoric acid and the iike. Salts derived from organic acids include, e.g., acetîc acid, propionic acid, gluconic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, ci trie acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluene-sulfonic acid, salicylic acid and the like. Likewise, pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases. Salts derived from inorganic bases include, by way of example only, sodium, potassium, lithium, aiuminutn, ammonium, calcium and magnésium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, such as alkyl amines (Le., NH2(alkyl)), dialkyl amines (Le,, HN(alkyl)2), trialkyl amines (i.e., N(alkyl)3), substituted alkyl amines (i.e., NH2(substituted alkyl)), di(substituted alkyl) amines (Le., HN(substituted alkyl)z), tri(substituted alkyl) amines (Le., N(substituted alkyl)s), alkenyl amines (Le., NH2(alkenyl)), dialkenyl amines (Le., HN(alkenyl)2), trîalkenyl

F amines (Le., N(alkenyl)}), substituted alkenyl amines (Le., NH2(substituted alkenyl)), di(substituted alkenyl) amines (Le., HN(substituted alkenyl);), tri(substiluted alkenyl) amines (Le., N(substituted alkenyl)3, mono-, di- or tri-cycloalkyl amines (Le., NFh/cycloalkyl), HN(cycloalkyl)2, N(cycloalkyl)3), mono-, di- or tri-arylamines (Le., NH;(aryl), HN(aryl)2, 5 N(aryl)3) or mixed amines, etc. Spécifie examples of suitable amines include, by way of example only, îsopropy lamine, trimethyl amine, diethyl amine, tri(iso-propyl) amine, trî(npropyl) amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, Nethylpiperidine and the like. In some embodiments, a pharmaceutically acceptable sait does not include a sait of a primary amine.

lü [0062] The term “hydrate” refers to the complex formed by the combining of a compound described herein and water.

[0063] A “solvaté” refers to an association or complex of one or more solvent molécules and a compound of the disclosure. Examples of solvents that form solvatés include, but are not limited to, water, isopropanol, éthanol, methanol, dimethylsulfoxide, ethylacetate, acetic acid and 15 ethanolamine.

[0064] Some of the compounds exist as tautomers. Tautomers are in equilibrium with one another. For example, amide containing compounds may exist in equilibrium with imidic acid tautomers. Regardless of which tautomer is shown and regardless of the nature of the equilibrium among tautomers, the compounds are understood by one of ordinary skill in the art 20 to comprise both amide and imidic acid tautomers. Thus, the amide containing compounds are understood to include their imidic acid tautomers. Likewîse, the imidic acid containing compounds are understood to include their amide tautomers.

[0065] The compounds of the invention, or their pharmaceutically acceptable salts include an asymmetric center and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino acids. The présent invention is meant to include ail such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (Æ)- and (S)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, for example, chromatography and fractîonal crystallization. Conventional techniques for the preparation/isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or the racemate of a sait or dérivative) using, for example, chiral high performance liquid chromatography (HPLC). When the compounds described herein contai n olefinîc double bonds or other centres of géométrie asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z géométrie isomers.

[0066] A “stéréoisomer” refers to a compound made up of the same atoms bonded by the same bonds but having different three-dimensional structures, which are not interchangeable. The présent invention contemplâtes various stereoîsomers and mixtures thereof and includes “enantiomers,” which refers to two stereoîsomers whose molécules are nonsuperimposeable mirror images of one another.

[0067] “Diastereomers” are stereoîsomers that hâve at least two asymmetric atoms, but which are not mirror-images of each other.

[0068] Relative centers of the compounds as depicted herein are indicated graphically using the “thick bond” style (bold or parallel lines) and absolute stereochemistry is depicted using wedge bonds (bold or parallel lines).

[0069] “Prodrugs” means any compound which releases an active parent drug aceording to a structure described herein in vivo when such prodrug is administered to a mammalian subject. Prodrugs of a compound described herein are prepared by modifying functional groups présent in the compound described herein in such a way that the modifications may be cleaved in vivo to release the parent compound. Prodrugs may be prepared by modifying functional groups présent in the compounds in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compounds. Prodrugs include compounds described herein wherein a hydroxy, amino, carboxyl, or sulfhydryl group in a compound described herein is bonded to any group that may be cleaved in vivo to regenerate the free hydroxy, amino, or sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to esters (e.g., acetate, formate and benzoate dérivatives), amides, guanidines, carbamales (e.g., N,Ndimethylaminocarbonyl) of hydroxy functional groups in compounds described herein and the like. Préparation, sélection and use of prodrugs îs dîscussed in T. Higuchi and V. Stella, “Prodrugs asNovel Delivery Systems,” Vol. 14 ofthe A.C.S. Symposium Sériés; “Design of Prodrugs,” ed. H. Bundgaard, Elsevier, 1985; and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, each of which are hereby incorporated by reference in their entirety.

[0070] The term, “métabolite,” as used herein refers to a resulting product formed when a compound disclosed herein is metabolized. As used hereîn, the term “metabolîzed” refers to the sum of processes (including but not limited to hydrolysis reactions and réactions catalyzed by enzymes) by which a particular substance, such as a compound disclosed herein, is changed by an organism. For example, an aldéhyde moiety (-C(O)H) of the compounds of the invention may be reduced in vivo to a -CH2OH moiety.

[0071] The tenu “hydroxy protecting group” refers to a Chemical moiety which is added to, and later removed from, a hydroxy functionality to obtain chemoselectîvity in a subséquent Chemical reaction. Exemplary protecting groups, as well as the methods for deprotection, 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 by acid), dimethoxytrityl or [bîs-(4-methoxyphenyl)phenylmethyl] (DMT) (removed by weak acid), methoxymethyl ether (MOM) (removed by acid), methoxytrityl or [(4methoxyphenyl)diphenylmethyl] (MMT) (removed by acid and hydrogenolysis), pmethoxybenzyl ether (PMB) (removed by acid, hydrogenolysis, or oxidation), methylthîomethyl ether (removed by acid), pivaloyl (Piv) (removed by acid, base or reductant agents), tetrahydropyranyl (THP) (removed by acid), tetrahydrofuran (THF) (removed by acid), trityl (triphenylmethyl, Tr) (removed by acid and hydrogenolysis), silyl ether (e.g., trimethylsily 1 (TMS), tert-butyldimethylsilyl (TBDMS), tri-iso-propylsilyloxymethyl (TOM), and triisopropylsilyl (TIPS) ethers) (removed by acid or fluorîde ion, such as NaF, TBAF (tetra-nbutylammonium fluoride, HF-Py, or HF-NEt3))„ methyl ethers (removed by cleavage is by TMSI in dichloromethane or acetonitrile or chloroform, or BBr3 in DCM), ethoxyethyl ethers (EE) (removed by IN hydrochloric acid).

Compounds

[0072] Provided herein are compounds that are useful as modulators of hemoglobin. It is contemplatcd that compounds disclosed herein hâve an improved pharmacokinetic profile relative to known modulators of hemoglobin whiie maintaining or improving efficacy. It is further contemplaled that compounds disclosed herein hâve an improved safety pharmacological profile relative to known modulators of hemoglobin.

[0073] Provided herein îs a compound of formula I:

I, or an isotopically enriched analog, stereoîsomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable sait of each thereof, wherein X, Y, Z, and R¹ are as defined herein.

[0074] In some embodiments, X is CH or N. In some embodiments, X is CH. In some embodiments, X is N.

[0075] In some embodiments, Y is CH or N. In some embodiments, Y is CH. In some embodiments, Y is N.

[0076] 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 îs N. In some embodiments, X is N; and Y is N.

[0077] In some embodiments, Z is absent, 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, CH2, or O. In some embodiments, Z is absent. In some embodiments, Z is CH2. In some embodiments, Z is O. In some embodiments, Z is S.

[0078] In some embodiments, R¹ is mono-hydroxy-(Ci.4 alkyl), di-hydroxy-(Ci-4 alkyl),

-CH2CH2OCH3, -CH2CH2CN, or ^x .In some embodiments, R¹ is mono-hydroxy(Ci-4 alkyl), di-hydroxy-(Ci-4 alkyl), -CH2CH2CN, or ' .In some embodiments, R¹ is mono-hydroxy-(Ci-4 alkyl), di-hydroxy-(Cw alkyl), -CH2CH2OCH3, or -CH2CH2CN. In some embodiments, R¹ is mono-hydroxy-(Ci-4 alkyl), di-hydroxy-(Ci-4 alkyl), or -CH2CH2CN.

[0079] In some embodiments, R¹ is -CH₂OH, -CH₂CH₂OH, -CH₂CH₂OCH₃, -CH₂CH₂CN, or . In some embodiments, R¹ is —CH₂OH, -CH₂CH₂OH, -CH₂CH₂CN, or ' . In some embodiments, R¹ is -CH₂OH, -CH₂CH₂OH, C^CIhOCIh, or

-CH₂CH₂CN. In some embodiments, R¹ is -CH2OH, -CH2CH2OH, or -CH2CH2CN. In some embodiments, R¹ is -CH2OH or -~CH₂CH₂OH. In some embodiments, R¹ is ~CH2OH. In some embodiments, R¹ is -CH2CH₂OH. In some embodiments, R¹ is -CH2CH2OCH3. In some embodiments, R¹ is -CH₂CH₂CN. In some embodiments, R¹ is

[0080] In some embodiments, R¹ is mono-hydroxy-(Ci-4 alkyl) or di-hydroxy-(CiM alkyl). In some embodiments, R¹ is mono-hydroxy-(Ci-4 alkyl). In some embodiments, R¹ is di-hydroxy(C2-4 alkyl). In some embodiments, R¹ is mono-hydroxy-(Ci-3 alkyl) or di-hydroxy-(Ci-3 alkyl). In some embodiments, R¹ is mono-hydroxy-(Ci-3 alkyl). In some embodiments, R¹ is dihydroxy-(C₂-3 alkyl). In some embodiments, R¹ is mono-hydroxy-(Ci-3 alkyl) or di-hydroxy-(Ci2 alkyl).

[0081] In some embodiments, R¹ is hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyI, 1,2dihydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, or 2-hydroxy-2-methylpropyl. In some embodiments, R¹ is hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 1,2-dihydroxyethyl, or 2hydroxypropyl. In some embodiments, R¹ is hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, or 2-hydroxypropyl.

[0082] In some embodiments, R¹ is hydroxymethyl (Le., -CH₂OH) or 2-hydroxyethyl (Le., —CH2CH2OH). In some embodiments, R¹ is hydroxymethyl.

[0083] In some embodiments, R¹ is 1-hydroxyethyl or 2-hydroxyethyl. In some embodiments,

OH QH

R¹ is 1-hydroxyethyl. In some embodiments, R¹ is In some embodiments, R¹ is

In some embodiments, R¹ is 2-hydroxyethyl. In some embodiments, R¹ is 1,2-dihydroxyethyl. In .OH .OH some embodiments, R¹ is °^H . In some embodiments, R¹ is .

[0084]

In some embodiments, R¹ is 2-hydroxypropyl. In some embodiments, R¹ is

In some embodiments, R¹ is

[0085] In some embodiments, R¹ is 3-hydroxypropyl.

[0086] In some embodiments, R¹ is 2-hydroxy-2-methylpropyl.

_R1a _R1c _R1₃ )—(~R^1b

[0087] In some embodiments, R¹ is ^OH or ^0H , wherein R^la is hydrogen or methyl; R^lb is hydrogen or methyl; and R^lc is hydrogen or hydroxy.

[0088] Any of the combinations of X, Y, Z, and R¹ are encompassed and provided by this disclosure.

[0089] Some embodiments provide for a compound of formula 1:

H

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

X is CH or N;

Y is CH or N;

Z is absent, CH2, O, or S; and

R¹ is -O bOH, -CH2CH2OH, -CH2CH2OCH3, -CH2CH2CN, or

[0090] Some embodiments provide for a compound of formula I:

or an isotopîcally enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable sait of each thereof, wherein:

X is CH or N;

Y is CH or N;

Z is absent, CH₂, or O; and

R¹ is -CH₂0H, -CH₂CH₂OH, -CIhCH₂OCH₃, -CH₂CH₂CN, or '

[0091] In some embodiments, Y is CH; and Z is CH₂.

[0092] Some embodiments provide for a compound of formula la:

O

la, or an isotopîcally enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable sait of each thereof, wherein:

X is CH or N; and

R¹ is -CHzOH, -CH₂CH₂OH, -CH2CH2OCH3, -CH₂CH₂CN, or

[0093] In some embodiments, X is CH or N; Y is CH; Z is CH₂, O, or S; and R¹ is a monohydroxy-(Ci-4 alkyl) or di-hydroxy-(Cm alkyl) moiety as described herein.

I [0094] In some embodiments, X is CH or N; Y is CH; Z is CHa, O, or S; and R¹ is a monohydroxy-(CM alkyl) moiety as described herein.

[0095] In some embodiments, X is N; Y is CH; Z is CH2, O, or S; and R¹ is a mono-hydroxy(C1-4 alkyl) moiety as described herein.

[0096] In some embodiments, X is N; Y is CH; Z is O or S; and R¹ is mono-hydroxy-(Ci-4 alkyl) moiety as described herein.

[0097] In some embodiments, X is N; Y is CH; Z is CH2, O, or S; and R¹ is -CH2OH or -CH2CH2OH. In some embodiments, X is N; Y is CH; Z is O or S; and R¹ is -CH2OH or

-CH2CH2OH. In some embodiments, X is N; Y is CH; Z is CH₂; and R¹ is -CH₂OH or

-CH2CH2OH. In some embodiments, X is N; Y is CH; Z is O; and R¹ is -CH2OH or

-CH2CH2OH. In some embodiments, X is N; Y is CH; Z is S; and R¹ is -CH2OH or

-CH2CH2OH.

[0098] Some embodiments provide for a compound of formula Ib:

Ib, or an isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable sait of each thereof, wherein:

Z is CHz, O, or S; and

R¹ is a mono-hydroxy-(Ci-4 alkyl) moiety as described herein.

[0099] In some embodiments, X is CH or N; Y is CH; Z is CH₂, O, or S; and R¹ is a di- hydiOxy-(C2-4 alkyl) moiety as described herein. In some embodiments, X is CH; Y is CH; Z is CH2, O, or S; and R¹ is a di-hydroxy-(C.2v alkyl) moiety as described herein. In some embodiments, X is CH; Y is CH; Z is O or S; and R¹ is a di-hydroxy-(C2-4 alkyl) moiety as described herein. In some embodiments, X is CH; Y is CH; Z is CH2; and R¹ is a di-hydroxy- (C2-4 alkyl) moiety as described herein. In some embodiments, X is CH; Y is CH; Z is O; and R¹ is a di-hydroxy-(C2-4 alkyl) moiety as described herein. In some embodiments, X is CH; Y is CH; Z is S; and R¹ is di-hydroxy-(C2-4 alkyl) moiety as described herein.

I

[0100] Sonie embodiments provide for a compound of formula le:

le, or an isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable sait of each thereof, wherein:

Z is CH2, O, or S; and

R¹ is a di-hydroxy-(Ci-4 alkyl) moiety as described herein.

[0101] Provided herein is a compound of formula;

or an isotopîcally enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or 5 a pharmaceutically acceptable sait of each thereof.

[0102] Provided herein is a compound of formula:

or an isotopically enriched anaiog, stereoisomer, mixture of stereoîsomers, or prodrug thereof, or a pharmaceutically acceptable sait of each thereof.

[0103] Provided herein is a compound of formula:

I

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

[0104] Provided herein is a compound of formula:

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

[0105]

Provided herein is a compound of formula:

or a pharmaceutically acceptable sait of each thereof.

[0106] In some embodiments, the compound is:

or a pharmaceutically acceptable sait thereof.

[0107] In some embodiments, the compound is:

[0108] In some embodiments, the compound is:

or a pharmaceutically acceptable sait thereof.

[0109] In some embodiments, the compound is:

[0110] In some embodiments, the compound is:

or a pharmaceutically acceptable sait thereof.

[OUI] In some embodiments, the compound is:

[0112] In some embodiments, the compound is:

or a pharmaceutically acceptable sait thereof.

[0113]

In some embodiments, the compound is:

[0114] In some embodiments, the compound is:

OH

or a pharmaceutically acceptable sait thereof.

[0115] In some embodiments, the compound is:

OH

[0116] In sonie embodiments, the compound is:

or a pharmaceutically acceptable sait thereof.

[0117] In some embodiments, the compound is:

[0118] In some embodiments, the compound is:

or a pharmaceutically acceptable sait thereof.

[0119]

In some embodiments, the compound is;

OH

H

[0120]

In some embodiments, the compound is;

or a pharmaceutically acceptable sait thereof.

[0121] In some embodiments, the compound is:

[0122] In some embodiments, the compound is:

H or a pharmaceutically acceptable sait thereof.

[0123] In some embodiments, the compound is:

[0124] In some embodiments, the compound is:

or a pharmaceutically acceptable sait thereof.

[0125]

In some embodiments, the compound is:

[0126] In some embodiments, the compound is:

or a pharmaceutically acceptable sait thereof.

[0127] In some embodiments, the compound is:

[0128] In some embodiments, the compound is:

OH

or a pharmaceutically acceptable sait thereof.

[0129] In some embodiments, the compound is:

OH

[0130] In some embodiments, the compound is:

OH

or a pharmaceutically acceptable sait thereof.

[0131] In some embodiments, the compound is:

OH

[0132] In some embodiments, the compound is:

OH

or a pharmaceutically acceptable sait thereof.

[0133] In some embodiments, the compound is:

OH

[0134] In some embodiments, the compound is:

OH

or a pharmaceutically acceptable sait thereof.

[0135] In some embodiments, the compound is:

OH

[0136] Provided herein is a compound selected from Table 1, or an isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable sait of each thereof. Provided herein is a compound selected from Table 1, or a pharmaceutically acceptable sait thereof. Provided herein is a compound selected from Table 1.

[0137] Provided herein is a compound selected from Table 2, or an isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable sait of each thereof. Provided herein is a compound selected from Table 2, or a pharmaceutically acceptable sait thereof. Provided herein is a compound selected from Table 2.

[0138] Compound numbers and IUP AC names of compounds described herein are summarized in Table 1 and Table 2.

Table 1

Compound Number	Structure	IUPAC naine
1	o=\ —\ Z , ο / \ I - O >=o 1 Z	(S)-2-hydroxy-6-((l-(2-(2hy droxy ethyl)nicoti noy l)piperid in2-yl)methoxy)benzaldehyde

Compound Number	Structure	IUPAC name
2	T O=< O /--y o—4 O --x \=O O. z y	(S)-2-hydroxy-6-((l-(2-(2- methoxyethyl)nicotînoyl)piperidin -2-yl)methoxy)benzaldehyde
3	I ° \ P >—< o—4 Z ( \ ^z \—\ zQ	(S)-3-(3-(2-((2-formyl-3- hydroxyphenoxy)methyl)piperidin e-l-carbonyl)pyridin-2- yl)propanenitrile
4	F i o=Ç o Q \—X V=o o	(S)-2-hydroxy-6-((1-(2-(2- (pyrrolidin-1- yl)ethyl)nicotinoyl)piperidin-2yl)methoxy)benzaldehyde
5	OH C^yNCp 0 ^0 O ότ^	(S)-2-hy droxy -6-((1 -(2(hydroxymethyl)benzoyl)piperidin -2-y l)methoxy )benzaldehy de

Compound Number	Structure	IUPAC naine
6	Z χ O / X i O )=o 1 T	(S)-2-hydroxy-6-((l -(2-(2hydroxyethyl)benzoyl)piperidin-2yl)methoxy)benzaldehyde
7	OH ° Ό 0 (V^n	(S)-2-hy droxy-6-(( 1-(3-(2hydroxyethyl)pyrazine-2carbonyl)piperidin-2yl)methoxy)bcnzaldehyde
8	\ z 0=^ Z—\ O / \ I 4 # ° ^°z oz >=o 1 □:	(S)-2-hydroxy-6-((4-(2-(2hy droxyethy l)nîcotinoy l)morpholi n-3-yl)methoxy)benzaldehyde
9	\ 0^99 ,___. ____/ 1 ο ο >ο 1 I	(S)-2-hydroxy-6-((l-(2-(2- hydroxyethyi)nicotinoyl)pyrrolidîn -2-yl)methoxy)benzaldehyde

Compound Number	Structure	IUPAC name
10 (Enantiomer 2)	.OH	2-hy droxy-6-((4-(2-(2- hydroxyethyl)nicotinoyl)thiomorp holîn-3-yl)methoxy)benzaldehyde
10 (Enantiomer 1)	.OH GG θνγ 0 kO O GG ^^OH	2-hydroxy-6-((4-(2-(2hydroxyethyl)nicotinoyl)thiomorp holin-3-yl)methoxy)benzaldehyde
11	OH .N, ) OÇÇ 0 ^0 0 Ôû	(S)-2-hy droxy -6-(( 1 -(2- (hydroxyniethyl)nicotinoyl)piperid in-2-yl)melhoxy)benzaldehyde
12	OH .N. J 0 k O GG	(S)-2-hy droxy -6-((4-(2- (liydroxymethyl)nicotinoyl)morph olin-3 -y l)methoxy)benzal dehy de
13 (Enantiomer 1)	OH fG rs θγγ ° % O GG ^OH	2-hydroxy-6-((4-(2- (hydroxymethyl)nicotînoyl)thiomo rpho!in-3- yl)methoxy)benzaldehyde

Compound Number	Structure	IUPAC naine
13 (Enantiomer 2)	G Z XX // O=( '—O z^ -0 ç y—O '—ω O >=o -1 z	2-hydroxy -6-((4-(2- (hydroxymethyl)nicotinoyl)thiomo rpholin-3- y l)methoxy )benzal dehy de
14	F T O=f O θ^ο-Ο —O ^z	(S)-2-hydroxy-6-((l-(2-(2methoxyethyl)benzoyl)piperîdin2-yl)methoxy)benzaldehyde
15	F T O=< O z < ' ^-z \—\ /^°	(S)-3-(2-(2-((2-formyl-3hydroxyphenoxy)methyl)pîperîdin e-1carbonyl)phenyl)propanenitrile
16	*0 Ο=Λ --\ Z^ O \=O F t	(S)-2-hydroxy-6-((l-(3 -(2- hydroxyethyl)picoîinoyl)piperidin2-yl)methoxy )benzaldehy de
17	nÇj ° Ό 0 ÔÔ	(S)-2-hydroxy-6-((l-(2-(2- (pyrrolidin-1- yI)ethyl)benzoyl)piperidin-2yl)methoxy)benzaldehyde

Compound Number	Structure	IUPAC name
18	OH OÇQ o L, 0 0 ^-^OH	(S)-2~hydroxy-6-((l-(2-(3hydroxypropyl)nicotinoyl)piperidi n-2-yl)methoxy)benzaldehyde
19	O °=/ξ ,___. Ί ο ο )=ο 1 χ'	(S)-2-hydroxy-6-((l-(3-(2hydroxyethyl)pyrazine-2carbonyl)pyrrolidin-2y I)methoxy)benzaldehy de
20	F τ ο=< ο —Ο Χ—Ζ \—, Ζ 7	(S)-2-hydroxy-6-((4-(2-(2methoxyelhyl)nicotinoyl)morpholi n-3-yl)methoxy)benzaldehyde
21	Ζ. \ζ \\ // ο=λ \—\ ζ—. ο = Ο \=ο 1 X	(S)-2-hydroxy -6-((4 -(3-(2hydroxyethyl)pyrazine-2carbony])morpholîn-3yl)methoxy)benzaldehyde
22	,ΟΗ θγγ ° LO 0 ό^π ^^ΟΗ	(S)-2-hydroxy-6-((4-(2-(2hy droxy ethy l)benzoyl)morphol i n3-yl)methoxy)benzaldehyde

Compound Number	Structure	IUPAC name
23	OH CÇG ° Ό O όΛ” '-''ΌΗ	(S)-2-hydroxy -6-((4-(2- (hy droxy methy l)benzoy l)morpholi n-3-yl)methoxy)benzaIdehyde
24	\ Z o=/\ z^ \ __/ Ί o— oz )=o -1 I	(S)-2-hydroxy-6-((l-(2-(2- methoxyethyl)nicotmoyl)pyrrolidi n-2-yl)methoxy)benzaldehyde
25	1 ro i'Vn 0 0 ιίτ/π	(S)-2-hy droxy-6-((1 -(3-(2methoxyethyl)pyrazine-2carbonyl)pyrrolidin-2yl)methoxy)benzaldehyde
26	OH 0 0 00 ^^OH	(S)-2-hydroxy-6-((l-(2- (hydroxyniethyl)benzoyl)pyrrolidi n-2-yl)methoxy)benzaldehyde
27	/ T o=< 0 —o ^-z ^=0 Z Z 'iï-A	(S)-2-hydroxy-6-((4-(3-(2methoxyethyl)pyrazine-2carbonyl)morpholin-3yl)methoxy )benzalde hy de

Compound Number	Structure	IUPAC name
28	Ίτ i o et	(S)-3-(3-(3-((2-formyl-3- hydroxyphenoxy)methyl)morpholi ne-4-carbonyl)pyridin-2yl)propanenitrile
29	z^ O— c # ° O 7=0 *	(S)-2-hydroxy-6-((4-(2-(2- methoxyethyl)benzoyl)morpholin3-yl)methoxy)benzaldehyde
30	<n °τγ 0 0 ό^π	(S)-3-(3-(2-((2-formyl-3bydroxyphenoxy)methyl)pyrrolidi ne-l-carbonyI)pyridin-2yl)propanenitrile
31	O=( \ — F~\ / z Ç '^O O )=o X Z	(S)-3-(2-(3-((2-formyl-3hy droxy phenoxy )methy l)morphol i ne-4- carbony l)pheny l)propanenitrile
32	yy O=A '--\ Γ—\ /—< ) z ç V- o O o Vo T x	(S)-3-(3-(3-((2-formyl-3hy droxy phenoxy )methyl)morpholi ne-4-carbonyl)pyrazin-2yl)propanenitrile

Compound Number	Structure	IUPAC name
33	OH ° % O ι/Ύ '''''OH	(S)-2-hy droxy-6-((4-(3(hydroxymethyl)pyrazine-2carbonyl)morpholin-3yl)methoxy)benzaldehyde
34	0 L < OH 0 OH J^,CHO L T ^^OH	2-(((2S)-l-(2-(1,2- dihydroxyethyl)benzoyl)piperidin2-yl)methoxy)-6- hydroxybenzaldehyde
34 (Diastereomer 1)	y-? Ι'-'ΌΗ0 Ό OH Â.CHO m	2-(((2S)-l-(2-(1,2- dihydroxyethyl)benzoyi)piperidin2-yl)methoxy)-6- hydroxybenzaldehyde
34 (Diastereomer 2)	yç ,Mh° ko OH X.CHO L I	2-(((2S)-l -(2-(1,2- dihydroxyethyl)benzoyl)piperidin2-yl)methoxy)-6- hy droxybenzal dehy de
35 (Diastereomer 1)	(<Ύ Ms ί OH° ° 0H Ml H ^^•OH	2-(((3R)-4-(2-(l,2dihydroxyethyl)benzoyl)thiomorp holin-3-yl)methoxy)-6hydroxybenzaldehyde

Compound Number	Structure	IUPAC name
35 (Diastereomer 2)	ΓΧ O A o ] OH θ 0 °H ΛΑ rï H	2-(((3R)-4-(2-(l,2dihydroxyethyl)benzoyl)thiomorp holin-3-yl)methoxy)-6hydroxybenzaldehyde
36	ÎI4 i] À 0 k Ç OH O OH À^CHO OL	2-{[(2S)-l-[2-(l,2- dihydroxyethyÎ)pyridine-3carbonyl]piperidin-2-yl]methoxy}6-hydroxybenzaldehyde
37	O=< O O-Z z> x O '—Z Vo zQ	(R)-2-hydroxy-6-((4-(2-(2- hydroxyethyl)nîcotinoyl)morpholi n-3-yl)methoxy)benzaldehyde
38	Çz O=( '--\ Z^ O gVO 1 O >=o 1	(R)-2-hydroxy-6-((l-(2-(2hy droxyethyl)nïcotinoy l)piperidin2-yl)melhoxy)benzaldehyde
39	o=<^-G 2-\ O Q.OJ O >=o 1 I	(S)-2-hydroxy-6-((4-(2-(2- hydroxy-2- methylpropyl)nicotinoyl)morpholi n-3-yl)methoxy)benzaldehyde

Compound Number	Structure	IDPAC name
40	O o=( ^-o O >=o T	2-hydroxy-6-((4-(2- (hydroxyinethyl)benzoyl)thîomorp holin-3-yl)methoxy)benzaldehyde
40 (Enantiomer 1)	OH r^s T 0 ^0 0 X.	2-hydroxy-6-((4-(2- (hydroxymethyl)benzoyl)thiomorp holin-3-yI)methoxy)benzaldehyde
40 (Enantiomer 2)	OH î^s ΑΧ θ	2-hydroxy -6-((4-(2- (hydroxymethyl)benzoyl)thîomorp holin-3-yl)methoxy)benzaldehyde
41 (Diastereomer 1)	HO < Y ( o	2-hydroxy-6-(((3S)-4-(2-(lhy droxy ethy l)nicotinoy l)morphol i n-3-yl)methoxy)benzaldehyde
41 (Diastereomer 2)	F t ° \ ? i z 1 Yo o \ / 10	2-hydroxy-6-(((3S)-4-(2-(lhy droxy ethy 1 )nicotinoy l)morpholi n-3-yl)methoxy)benzaldehyde

I

Compound Number	Structure	IUPAC name
42 (Diastereomer 1)	\.OH .N. J if A Γ 0 CÇG ° LO 0 ^^OH	2-hydroxy-6-(((3S)-4-(2-(2hydroxypropyl)nicotînoyl)morphol in-3-yi)methoxy)benzaldehyde
42 (Diastereomer 2)	\.OH .KL J c T O 0 0 cû	2-hydroxy-6-(((3S)-4-(2-(2hydroxypropyl)nicotinoyl)morphol in -3 -y l)methoxy)benzaldehy de
43 (Diastereomer 1)	HO aYrs ^Λγ ° Ό 0 CC ^^OH	2-hydroxy-6-(((3 R)-4-(2-(lhydroxyethyl)nicotinoyl)thiomorp holin-3-yl)meîhoxy)benzaldehyde
43 (Diastereomer 2)	Y / \^O °Y y nrO ο Yo -11 -Z	2-hydroxy-6-(((3R)-4-(2-(l- hydroxyethyi)nicotinoyl)thiomorp holin-3-yl)methoxy)benzaldehyde
44 (Diastereomer 1)	F τ ογ Ο /^° Ό I \ / O H )-\ )^° O	2-hydroxy-6-(((3R)-4-(2-(2hydroxypropyl)nicotinoyl)thiomor pholin-3- yl)methoxy)benzaldehyde ___________________ _

I

Compound Number	Structure	1DPAC name
42 (Diastereomer 1)	Çz '—\ Z—, O __/ \ X C O I >=/ O >o -11 I	2-hydroxy-6-(((3S)-4-(2-(2hydroxypropyl)nicotinoyl)morphol in-3-yl)methoxy)benzaldehyde
42 (Diastereomer 2)	/ £ ° \ /° x O —z )—\ O	2-hydroxy-6-(((3S)-4-(2-(2hydroxypropyl)nicotinoyl)morphol in-3-yl)methoxy)benzaldebyde
43 (Diastereomer 1)	HO îi rs ^ίίΝΎ^ 0 k O	2-hydroxy-6-(((3R)-4-(2-(lhydroxyethyl)nicotinoyl)thiomorp holin-3-yl)methoxy)benzaldehyde
43 (Diastereomer 2)	1 T X ° \ /° CO—C'A # | '--Z 1 )=O o \_/ x / A	2-hydroxy-6-(((3R)-4-(2-(lhydroxyethyl)nicotînoyl)thiomorp holin-3-yl)methoxy)benzaldehyde
44 (Diastereomer 1)	\ ,oh <v Cs cvy 0 k0 0 _______^-^0 H___	2-hydroxy-6-(((3R)-4-(2-(2hy droxy propy l)nicotinoy l)thîomor pholin-3- yl)methoxy)benzaldehyde

Compound Number	Structure	IUPAC name
44 (Diastereomer 2)	O1 O=\ \—ζ z x O / \ z ç 'w— ο O >=o 1 z'	2-hydroxy-6-(((3R)-4-(2-(2hydroxypropyl)nicotinoyl)thiomor pholin-3- yl)methoxy)benzaldehyde

Table 2

[0139] Provided herein is a compound selected from Table 6, or an isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable sait of each thereof, and excluding Référencé Compound A, B, and C. Provided herein is a compound selected from Table 6, or a pharmaceutically acceptable sali thereof, and excluding Référencé Compound A, B, and C. Provided herein is a compound selected from Table 6 and excluding Référencé Compound A, B, and C.

[0140] Provided herein is a compound selected from Table 7, or an isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable sait of each thereof, and excluding Reference Compound A and B. Provided herein is a compound selected from Table 7, or a pharmaceutically acceptable sait thereof, and excluding Reference Compound A and B. Provided herein is a compound selected from Table 7 and excluding Reference Compound A and B.

Treatment Methods and Uses

[0141] “Treatment” or “treating” is an approach for obtaining bénéficiai or desired results including clinîcal results. Bénéficiai or desired clînical results may include one or more of the following; a) inhibiting the disease or condition (e.g., decreasing one or more symptoms resulting from the disease or condition, and/or diminishing the extent of the disease or condition); b) slowing or arresting the development of one or more clinîcal symptoms associated with the disease or condition (e.g., stabîlizing the disease or condition, preventing or delaying the worsenîng or progression of the disease or condition, and/or preventing or delaying the spread (e.g., metastasis) of the disease or condition); and/or c) relieving the disease, that is, causing the régression of clinîcal symptoms (e.g., ameliorating the disease State, providing partial or total remission of the disease or condition, enhancing effect of another médication, delaying the progression of the disease, increasing the quality of life, and/or prolonging survival.

[0142] “Prévention” or “preventing” means any treatment of a disease or condition that causes the clinîcal symptoms of the disease or condition not to develop. Compounds may, in some embodiments, be administered to a subject (including a human) who is at risk or has a family 20 history of the disease or condition.

[0143] “Subject” refers to an animal, such as a mammal (including a human), that has been or will be the object of treatment, observation or experiment. The methods described herein may be useful in human therapy and/or veterinary applications. In some embodiments, the subject is a mammal. In one embodiment, the subject is a human.

[0144] The term “therapeutically effective amount” or “effective amount” of a compound described herein or a pharmaceutically acceptable sait, tautomer, stereoisomer, mixture of stereoîsomers, prodrug, or deuterated analog thereof means an amount suffîcient to effect treatment when administered to a subject, to provide a therapeutic benefit such as amelioration of symptoms or slowing of disease progression. For example, a therapeutically effective amount may be an amount suffîcient to decrease a symptom of a sickle cell disease. The therapeutically effective amount may vary depending on the subject, and disease or condition being treated, the weight and âge of the subject, the severity of the disease or condition, and the manner of administering, which can readily be determined by one of ordinary skill in the art.

[0145] The methods described herein may be applied to ceil populations in vivo or ex vivo. “In vivo” means within a living individual, as within an animal or human. In this context, the methods described herein may be used therapeutically in an individual. “Ex vivo” means outsîde of a living individual. Examples of ex vivo cell populations include in vitro cell cultures and biological samples including fluid or tissue sam pi es obtained from individuals. Such samples may be obtained by methods well known in the art. Exemplary biological fluid samples include blood, cerebrospinal fluid, urine, and saliva. In this context, the compounds and compositions described herein may be used for a variety of purposes, including therapeutic and experimental purposes. For example, the compounds and compositions described herein may be used ex vivo to détermine the optimal schedule and/or dosing of administration of a compound of the présent disclosure for a given indication, cell type, individual, and other parameters. Information gleaned from such use may be used for experimental purposes or in the clinîc to set protocols for in vivo treatment. Other ex vivo uses for which the compounds and compositions described herein may be suited are described below or wîll become apparent to those skilled in the art. The selected compounds may be further characterized to examine the safety or tolérance dosage in human or non-human subjects. Such properties may be examined using commonly known methods to those skilled in the art.

[0146] The term “hemoglobin” as used herein refers to any hemoglobin protein, including normal hemoglobin (HbA) and abnormal hemoglobin, such as sickle hemoglobin (HbS).

[0147] The term “sickle cell disease” refers to diseases mediated by sickle hemoglobin (HbS) that results from a single point mutation in the hemoglobin (Hb). Sickle cell diseases include sickle cell anémia (HbSS), hemoglobin SC disease (HbSC), hemoglobin S beta-plus-thalassemia (HbS/p+) and hemoglobin S beta-zero-thalassemia (HbS/βΟ).

[0148] Provided herein are methods for treating sickle cell disease (SCD). Sickle hemoglobin (HbS) contains a point mutation where glutamic acid is replaced with valine, making HbS susceptible to polymerization under hypoxie conditions to give the HbS containing red blood cells their characteristic sickle shape. The sickled cells are also more rigid than normal red blood cells, and their lack of flexibîlity can lead to blockage of blood vessels. It is contemplated that an approach to therapy would be to maintain the HbS in the oxygenated State, as polymerization occurs only in the deoxygenated State under hypoxie conditions.

solvents, perméation enhancers, solubilizers and adjuvants. Such compositions are prepared in a manner well known in the pharmaceutical art. See, e.g., Remington’s Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa. 17th Ed. (1985); and Modem Pharmaceutics, Marcel Dekker, Inc. 3rd Ed. (G.S. Banker & C.T. Rhodes, Eds.).

[0154] The pharmaceutical compositions may be administered in either single or multiple doses. The pharmaceutical composition may be administered by various methods încluding, for example, rectal, buccal, intranasal and transdermal routes. In certain embodiments, the pharmaceutical composition may be administered by intra-arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, topically, or as an inhalant.

[0155] One mode for administration is patenterai, for example, by injection. The forms in which the pharmaceutical compositions described herein may be incorporated for administration by injection include, for example, aqueous or oil suspensions, or émulsions, with sesame oil, corn oil, cottonseed oil, or peanut oil, as well as élixirs, mannitol, dextrose, or a stérile aqueous 15 solution, and similar pharmaceutical vehicles.

[0156] Oral administration may be another route for administration of the compounds described herein. Administration may be via, for example, capsule or enteric coated tablets. In making the pharmaceutical compositions that include at least one compound described herein or an isotopically enriched analog, stereoisomer, mixture of stereoîsomers, or prodrug thereof, or a 20 pharmaceutically acceptable sait of each thereof, the active ingrédient is usually diluted by an excipient and/or enclosed within such a carrier that can be in the form of a capsule, sachet, paper or other container. When the excipient serves as a diluent, ît can be in the form of a solid, semisolid, or liquid material, which acts as a vehîcle, carrier or medium for the active ingrédient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, 25 élixirs, suspensions, émulsions, solutions, syrups, aérosols (as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatîn capsules, stérile injectable solutions, and stérile packaged powders.

[0157] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium 30 silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, stérile water, syrup, and methyl cellulose. The formulations can additionally include lubricating agents such as talc, magnésium stéarate, and minerai oil; wetting agents; emulsifying and suspending agents;

preserving agents such as methyl and propylhydroxy-benzoates; sweetening agents; and flavorîng agents.

[0158] The compositions that include at least one compound described herein or an isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a 5 pharmaceutically acceptable sait of each thereof can be formulated so as to provide quick, sustained or delayed release of the active ingrédient after administration to the subject by employing procedures known in the art. Controlled release drug delivery Systems for oral administration include osmotic pump Systems and dissolutional Systems containing polymercoated réservoirs or drug-polymer matrix formulations. Examples of controlled release Systems 10 are given in U.S. Patent Nos. 3,845,770; 4,326,525; 4,902,514; and 5,616,345. Another formulation for use in the methods disclosed herein employ transdermal delivery devices (“patches”). Such transdermal patches may be used to provide continuous or discontinuous infusion of the compounds described herein in controlled amounts. The construction and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. See, e.g., 15 U.S. Patent Nos. 5,023,252, 4,992,445 and 5,001,139. Such patches may be constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents.

[0159] For preparing solid compositions such as tablets, the principal active ingrédient may be mixed with a pharmaceutical excipient to form a solid preformulation composition containing a hoinogeneous mixture of a compound described herein or an isotopically enriched analog, 20 stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable sait of each thereof. When referring to these preformulation compositions as homogeneous, the active ingrédient may be dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules.

[0160] The tablets or pills of the compounds described herein may be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action, or to protect from the acid conditions of the stomach. For example, the tablet or pill can include an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer that serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodénum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate.

[0161] Compositions for inhalation or insufflation may include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders. The liquid or solîd compositions may contain suitable pharmaceutically acceptable excipients as described 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 may be nebulized by use of inert gases. Nebulized solutions may be inhaled directly from the nebulizîng device or the nebulizing device may be attached to a facemask tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered, preferably orally or nasally, from devices that deliver the formulation in an appropriate manner.

Dosing

[0162] The spécifie dose level of a compound of the présent application for any particular subject will dépend upon a variety of factors including the activity of the spécifie compound employed, the âge, body weight, general health, sex, diet, time of administration, route of administration, and rate of excrétion, drug combination and the severity of the particular disease in the subject undergoing therapy. For example, a dosage may be expressed as a number of milligrams of a compound described hereîn per kilogram of the subject’s body weight (mg/kg). Dosages of between about 0.1 and 150 mg/kg may be appropriate. In some embodiments, about 0.1 and 100 mg/kg may be appropriate. In other embodiments a dosage of between 0.5 and 60 mg/kg may be appropriate. Normalizîng according to the subject’s body weight is particularly useful when adjusting dosages between subjects of widely disparate sîze, such as occurs when using the drug in both children and adult humans or when converting an effective dosage in a non-human subject such as dog to a dosage suitable for a human subject.

Synthesis of the Compounds

[0163] The compounds may be prepared using the methods disclosed herein and routine modifications thereof, which will be apparent given the disclosure herein and methods well known in the art. Conventional and well-known synthetic methods may be used in addition to the teachings herein. The synthesis of typical compounds described hereîn may be accomplished as described in the following examples. If available, reagents may be purchased commercially, e.g., from Sigma Aldrich or other chemical supplîers.

General Synthesis

[0164] Typîcal embodiments of compounds described herein may be synthesized using the general reaction schemes described below. It will be apparent given the description herein that the general schemes may be altered by substitution of the starting materials with other materials having similar structures to resuit in products that are correspondingly different. Descriptions of synthèses follow to provide numerous examples of how the starting materials may vary to provide corresponding products. Given a desired product for which the substituent groups are defined, the necessary starting materials generally may be determined by inspection. Starting materials are typically obtained from commercial sources or synthesized using published methods. For synthesizing compounds which are embodiments described in the présent disclosure, inspection of the structure of the compound to be synthesized will provide the identity of each substituent group. The identity of the final product will generally render apparent the identity of the necessary starting materials by a simple process of inspection, given the examples herein. In general, compounds described herein are lypically stable and isolatable at room température and pressure.

[0165] In some embodiments, a compound of formula I can be synthesized by exemplary synthetic pathways as shown in Schemes A and B.

[0166] In some embodiments of Scheme A, R² can be hydroxyl or chloro; R¹ can be monohydroxy-(Cj-4 alkyl), CH2CH2OCH3, -CH2CH2CN, or 1 and X, Y, and Z are as described herein. As shown in Scheme A, compound Al and compound A2 are coupled first utilizing standard coupling conditions to give compound A3, which can be then assembled onto 2,6-dihydroxybenzaldehyde A4 to produce compound of formula I. In some embodiments, when R¹ is mono-hydroxy-(Ci-4 alkyl) of a compound of formula I, the hydroxy group of the

R¹ moiety of Al includes a hydroxy protecting group known in the art; the protecting group may be subsequently removed after coupling A3 to A4 utilizing standard procedures, thereby producing a compound of formula I.

[0167] In some embodiments of Scheme B, R³ can be a Cs^alkene; R² can be hydroxyl or chloro; R¹ can be di-hydroxy-(C2-4 alkyl); Q is a halo; PG is a hydroxy protecting group; and X, 5 Y, and Z are as described herein. As shown in Scheme B, compound B1 and compound B2 are cou pied first utilizing standard coupling conditions to give compound B3. Standard deprotection procedures provides compound B4, which can be then assembled onto 2,6dîhydroxybenzaldehyde A4 to produce compound B5. Introduction of the alkene (e.g. via Stilie coupling) provides compound B6, which can then be converted to a compound of formula I via 10 dihydroxylation procedures known in the art.

EXAMPLES

[0168] The following examples are included to demonstrate spécifie embodiments of the disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques to function wel! in the practice of the disclosure, and thus can be considered to constitute spécifie modes for its practice, However, those of skill in the art should, in iight of the présent disclosure, appreciate that many changes can be made in the spécifie embodiments which are disclosed and still obtain a like or similar resuit without departîng from the spirit and scope of the disclosure.

Synthetic Examples

Example 1. Synthesis of (S)-2-hydroxy-6-((l-(2-(2-hydroxy ethyl) nicotinoyl) piperidin-2y])methoxy)benzaldehyde, Compound 1

[0169] Compound 1 was synthesîzed according to Scheme IA.

Scheme IA

Steps 1-4

Step 5

1b (R = H, X = Br)

1c (R = TBS, X = Br)

1d (R = TBS, X = COOMe)

1e (R = TBS, X = COOH)

Steps 6&7

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

[0170] Into a 100-mL 3-necked round-bottom flask, was placed a solution of ethyl 2-(3bromopyridin-2-yl)acetate (la) (4 g, 16.39 mmol, 1 equiv) in tetrahydrofuran (“THF”) (40 mL).

This was followed by the addition of diisobutylaluminum hydride (“DIBAL-H”) in THF (16 mL, 32.00 mmol, 1.95 equiv) dropwise with stirring at -78 “C. The resulting mixture was allowed to warm to rt and was stirred for additlonal 3 hr at 25 “C. The reaction was then quenched by the addition of 50 mL of saturated NH4CI. The resulting solution was extracted with 3x100 mL of ethyl acetate and the organic layers combined, washed with 2x100 mL of brine The separated organic layer was dried over Na2SO4, active carbon, filtered and then concentrated. The residue was applied onto a silica gel column and eluted with ethyl acetate/petroleum ether (1:3) to provide the title compound. LCMS (ES) [M+l]⁺ m/z 202.0.

Step 2: Synthesis of 3-bromo-2-(2-((tert-butyhlimethylsilyl)oxy)ethyl)pyridine (le).

[0171] Into a 100-mL round-bottom flask, was placed a solution of 2-(3-bromopyridin-25 yl)ethan-l-ol (1.9 g, 9.40 mmol, 1 equiv) in dimethylformamide (“DMF) (20 mL), 1Hîmidazole (1.3 g, 18.81 mmol, 2 equiv), 4-dimethylaminopyridine (“DMAP”) (0.1 g, 0.94 mmol, 0.1 equiv), tert-butyl(chloro)dimethylsilane (2.8 g, 18.81 mmol, 2 equiv). The resulting solution was heated to 50 °C and stirred for 2 hr. The reaction mixture was cooled and extracted with 2x50 mL of ethyl acetate. The combined organic layers was washed with 2x50 mL of brine, drîed over NaiSOi, filtered and concentrated. The residue was applied onto a silica gel column and eluted with ethyl acetate/petroleum ether (1:3) to provide the title compound. LCMS (ES) [M+l]⁺ m/z 316.1.

[0172] Alternatively, a teri-butyldiphenylsilyl (TBDPS) protecting group can be used instead of ieri-butyl(chloro)dimethylsilyl (TBS). In typical conditions, îmidazole (1.5 to 4 eq) and tert15 butyl(chloro)dîphenylsilane (TBDPSC1 (about 1 eq.) were added to a solution of alcohol 1b (1 eq) in DCM (3 to 15 V). The reaction mixture was stirred at RT for 1 to 48 hours. This gave the product (lc2) after normal workup and purification. The TBDPS group can be removed using TBAF (1-3 eq) following typical literature conditions. Compounds 8 and 12 can be synthesized using TBDPS as a protecting group.

Step 3: Synthesis of methyl 2-(2-(( tert-butyldimethylsilyl)oxy)ethyl)nicotinate (ld).

[0173] Into a 250-mL sealed tube, was placed a solution of 3-bromo-2-[2-[(tertbutyldimethylsilyl)oxy] ethyl]pyridine (2.0 g, 6.32 mmol, 1 equiv) in methanol (“MeOH,” 100 mL), triethylamine (“TEA,” 1.3 g, 12.65 mmol, 2 equiv), and [1,1’bis(diphenylphosphino)ferrocene]dichloropalladium(II) (“Pd(dppf)Ch,” 0.5 g, 0.63 mmol, 0.1 equiv). The resulting solution was stirred for 16 hr at 100 °C under CO atmosphère (10 atm). After cooling to rt, the reaction mixture was filtered and the filtrate was concentrated. The resulting residue was applied onto a silica gel column and eluted with ethyl acetate/petroleum ether (1:3) to provide the title compound. LCMS (ES) [M+l]⁺ m/z 296.2.

Alternative Synthesis: Scheme IB

1c2

Scheme IB

[0174] Alternatively, phenyl formate can be used to replace CO gas as a carbonyl source, in the presence of triethylamine (2 eq), catalytic amounts of palladium acetate (e.g., 0.02 eq) and tri-terl-butylphosphonium tetrafluoroborate (e.g., 0.08 eq), to convert the bromide lc2 into carboxylate ld2 in acetonitrile (3 to 10 V) under heating (80 °C) for 2 to 48 hours, and then directly to carboxylic acid le2 by hydrolysis of the ester under basic aqueous conditions (K2CO3 2-8 eq in 3 to 10 V water; 50 to 80 °C for up to 48 hours).

Step 4: Synthesis of 2-(2-((tcrt-butyldimethylsilyl)oxy)ethyl)nicotinic acid (le).

[0175] Into a 100-mL round-botlom flask, was placed a solution of methyl 2-[2-[(tertbutyldimethylsilyl) oxy]ethyl]pyridine-3-carboxyIate (1.7 g, 5.75 mmol, 1 equiv) in MeOH (20 mL), and a solution of LiOH (275.6 mg, 11.51 mmol, 2 equiv) in H2O (5 mL). The resulting solution was stirred for 4 hr at 25 °C. Water (10ml) was added to the reaction mixture, crude product as precipitate was collected by filtration. The crude product was then purified by FlashPrep-HPLC using the following conditions (IntelFlash-1): Column, Cl8 silica gel; mobile phase, H2O:acetonitrile (“ACN”)=10:l increasingto H2O:ACN=3:1 within 10 minto provide the title compound. LCMS (ES) [M+l]⁺ m/z 282.1.

Step 5. Synthesis of (S)-(2-(2-((tert-bntyldimethylsilyl)oxy)ethyl)pyridin-3-yl)(2 20 (hydroxymethyl)piperidin-l-yl)methanone (If).

[0176] Into a 100-mL round-bottom flask, was placed 2-[2-[(tertbutyldimethylsilyl)oxy]ethyl]pyridine-3-carboxylic acid (1.4 g, 4.97 mmol, 1 equiv), [(2S)piperidin-2-yl]methanol (0.9 g, 7.46 mmol, 1.5 equiv), A, A-diisopropylethy lamine (“DIEA,” 1.3 g, 9.95 mmol, 2 equiv), l-[Bis(dimethylamino)methylene]-lH-l,2,3-triazolo[4,5-b]pyridinium

3-oxid hexafluorophosphate (“HATU,” 2.8 g, 7.46 mmol, 1.5 equiv) and 30 mL of dichloromethane (“DCM”). The resulting reaction mixture was stirred for 2 hr at 25 °C and then diluted with 60 mL of H₂O. The organic phase was extracted with 3x50 mL of ethyl acetate. The combinée! organic layers was drîed over Na2SÛ4, filtered and concentrated. The residue was applied onto a silica gel column and eluted with ethyl acetate/petroleum ether (1:3) to provide the title compound. LCMS (ES) [M+l]⁺ m/z 379.2.

Steps 6 & 7: Synthesis of (S)-2-hydroxy-6-((l-(2-(2-hydroxyethyl)nicotinoyI)piperidin-2yl)methoxy)benzaldehyde (Compound 1).

[0177] Into a 100-mL round-bottom flask, was placed a solution of [(2S)-1-(2-[2-[(tertbutyldimethylsilyl)oxy]ethyl]pyridine-3-carbonyl)piperidin-2-yl]methanol (900 mg, 2.38 mmol, 1 equiv) in DCM (8 mL), 2,6-dîhydroxybenzaldehyde (656.7 mg, 4.75 mmol, 2 equiv), and trîphenylphosphine (“PPh3,” 1247.0 mg, 4.75 mmol, 2 equiv). This was followed by the addition of a solution of di-fôri-butyl azodicarboxylate (“DBAD,” 1094.8 mg, 4.75 mmol, 2 equiv) in DCM (2 mL) dropwise with stirring at 0 °C. The resulting solution was stirred for 2 hr at 25 °C. The reaction mixture was concentrated and dîssolved in 20 mL THF. To this was added tetrabutylammonium fluoride (“TB A F,” 1243.1 mg, 4.75 mmol, 2 equiv). The resulting mixture was allowed to stir for 2 hr at 25 °C. The reaction mixture was concentrated to give a crude product, which was purified by Prep-HPLC with the following conditions (Prep-HPLC-006): Column, XBridge Prep C18 OBD Column, 19mm x 150mm 5um; mobile phase, Water(10 mmoL/L NH4HCO₃+0.1%NH3.FI₂O) and ACN (14% Phase B up to 35% in 8 min, hold 95% in 1 min, down to 14% in 1 min, hold 14% in 1 min); Detector, UV 254 nm. This provided the title compound. ^]HTEM NMR (300 MHz, 353K, dimethylsuifoxide (“DMSO”)-î/₆): S 11.36 (s, 1H), 10.25 (s, 1H), 8.51 (dd, J = 4.8,1.8 Hz, 1H), 7.51-723 (m, 3H), 6.7-6.5 (m, 2H), 5.15 (s, 1H), 4.59 - 3.98 (m, 3H), 3.78 (br, 2H), 3.17-2.86 (m, 4H), 1.83-1.37 (m, 6H). LCMS (ES) [M+l]⁺ m/z 385.2.

Example 2: (S)-2-hydroxy-6-((l-(2-(2-methoxyethyl)nicotinoyl)piperidin-2yl)methoxy)benzaldehyde, Compound 2

[0178] Compound 2 was synthesized according to Scheme 2.

Scheme 2

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

[0179] Into a 100-mL round-bottom flask, was placed a mixture of methyl 2-chIoropyridine-3carboxylate (3 g, 17.48 mmol, 1.00 equiv), dioxane (40 mL), water (4 mL), 2-ethenyl-4,4,5,5tetramethyl-l,3,2-dioxaborolane (5.39 g, 34.99 mmol, 2.00 equiv), CssCOs (11.40 g, 34.99 mmol, 2.00 equiv) and tetrakis(triphenylphosphine)pal!adium(0) (“Pd(PPh3)4,” 2.02 g, 1.75 mmol, 0.10 equiv). The resulting solution was stirred for 2 h at 100 °C under N2. The reaction mixture was cooled, filtered, and conceutrated under vacuum. The resulting residue was purified by a silica gel column by eluting with ethyl acetate/petroleum ether (1/2) to give 2 methyl 2ethenylpyrîdine-3-carboxylate. LCMS (ES) [M+l]⁺ m/z: 164.1.

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

[0180] Into a 100-mL round-bottom flask, was placed a solution of methyl 2-ethenylpyridine3-carboxylate (1.5 g, 9.19 mmol, 1.00 equiv), methanol (20 mL) and aqueous hydrogen chlorîde ι

(36%, 2 mL). The resulting solution was stirred for 48 h at 60 °C. The mixture was cooled and then concentrated under vacuum to give melhyl 2-(2-methoxyethyl)pyridine-3~carboxylate). LCMS (ES) [M+l]⁺ m/z: 196.1.

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

[0181J Into a 100-mL round-bottom flask, was placed a solution of methyl 2-(2methoxyethyl)pyridinc-3-carboxylate (2.50 g, 12.81 mmol, 1.00 equiv), methanol (30 mL), H₂O (6 mL) and NaOH (2.56 g, 64.00 mmol, 5.00 equiv). The resulting solution was stirred for 2 h at 50 °C. The reaction was cooled, pH adjusted to 6 with addition of aqueous hydrogen chloride (2 M). The mixture was extracted with 3x50 mL of DCM/MeOH (10/1). The combined organic layers was washed with 1x100 mL of brine, dried over anhydrous sodium sulfate and concentrated under vacuum to produce (crude) 2-(2-methoxyethyl)pyridine-3-carboxylic acid. LCMS (ES) [M+l] ⁺ m/z: 182.1.

Step 4: Synthesis of [(2S)-l-[[2-(2-methoxyethyl)pyridin-3-yl]carbonyI]piperidin-2yl]methanol (2e).

[0182] Into a 100-mL round-bottom flask, was placed a solution of 2-(2methoxyethyl)pyridine-3-carboxylic acid (600 mg, 3.31 mmol, 1.00 equiv), dichloromethane (30 mL), (2S)-piperidin-2-ylmethanol (762 mg, 6.62 mmol, 2.00 equiv), DIEA (855 mg, 6.62 mmol, 2.00 equiv) and HATU (1.89 g, 4.97 mmol, 1.50 equiv). The resulting solution was stirred for 1 h at rt. The crude reaction mixture was filtered and concentrated. The resulting residue was purified by reverse préparative HPLC (Prep-C18, 20-45 μΜ, 120 g, Tianjin Bonna-Agela Technologies; gradient elution of 15% CH3CN in water to 40% CH3CN in water over a 12 min period, where both solvents contain 0.1% ammonia) to provide [(2S)-l-[[2-(2methoxyethyl)pyridin-3-yl]carbonyl]piperidin-2-yl]melhanoL LCMS (ES) [M+l] ⁺ m/z: 279.1.

Step 5: (S)-2-hydroxy-6-((l-(2-(2-methoxyethyl)nicotinoyl)piperidin-2 25 yl)methoxy)benzaldehyde (Compound 2).

[0183] Into a 50-mL 3-necked round-bottom flask, was placed a solution of [(2S)-l-[[2-(2methoxyethyl)pyridin-3-yl]carbonyl]piperidin-2-yl]methanol (265 mg, 0.95 mmol, 1.00 equiv), dichloromethane (10 mL), 2,6-dihydroxybenzaldehyde (263 mg, 1.90 mmol, 2.00 equiv) and PPh₃ (499 mg, 1.90 mmol, 2.00 equiv). It was added the solution of dibenzyl azodîcarboxylate (“DBAD”) (438 mg, 1.90 mmol, 2.00 equiv) in DCM (5 mL) under N₂ at 0°C. The resulting solution was stirred for 2 h at room température. The resulting mixture was concentrated under vacuum. The residue was purifîed by a sîlica gel column eluted wîth ethyl acetate/petroleum ether (1/1). The crude reaction mixture was filtered and subjected to reverse préparative HPLC (Prep-C18, 5 mM XBridge column, 19 x 150 mm, waters; gradient elution of 22% CHsCN in water to 42% CH3CN in water over a 6 min period, where bolh solvents contain 0.1% trifluoroacetic acid (“TFA”)) to provide 2-hydroxy-6-[(l-[hydroxy[2-(2-methoxyethyl)piperidin3-yl]methyi]piperidin-2-yl)methoxy]cyclohexane-l-carbaldehyde. ^]ΗΤΕΜ NMR (300 MHz, 353 K, DMSO-Jc) 5 11.64(s, 1H), 10.28(s, 1H), 8.56(dd, 7=5.1 Hz, 1.8 Hz, 1H), 7.78-7.59(m, 1H), 7.52(t, 7=8.1 Hz, 1H), 7.39-7.21(m, 1H), 6.72(s, 1H), 6.55(d, 7=8.4 Hz, lH),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+l]⁺ m/z: 399.1.

Example 3: (S)-3-(3-(2-((2-formyI-3-hydroxyphenoxy) methyl) piperidine-lcarbonyl)pyridin-2-yl)propanenitrile, Compound 3

[0184] Compound 3 was synthesized according to Scheme 3.

Scheme 3

Step 1: Synthesis of methyl 2-formylnicotinate (3 b).

[0185] Into a 100-mL round-bottom flask, was placed a solution of methyl 2-methylpyridine3-carboxyIate (5 g, 33.08 mmol, 1 equiv) in dioxane (50 mL), (oxo-lambda4selanylîdene)oxidane (sélénium dioxide) (5.5 g, 49.61 mmol, 1.5 equiv). After stirringfor 16 hr 5 at 110 °C the reaction mixture was cooled to rt, concentrated, and diluted with 100 mL of H2O.

It was then extracted with 4x100 ml of ethyl acetate and the organic layers combined. The organic layers was washed with 200 ml of brine, dried over Na₂SO4 and concentrated. The resulting residue was applied onto a silica gel column and eluted with ethyl acetate/petroleum ether (1:5) to provide the tîtle compound. LCMS (ES) [M+l]⁺ m/z 166.0.

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

[0186] Into a 100-mL round-bottom flask, was placed a solution of methyl 2-formylpyridine-3carboxylate (2.5 g, 15.14 mmol, 1 equiv) in THF (30 mL). This was followed by the addition of diethyl (cyanomethyl)phosphonate (3.2 g, 18.17 mmol, 1.2 equiv) at 0 °C and (tertbutoxy)potassium (2.5 g, 22.71 mmol, 1.5 equiv), in portions at 0 °C. The resulting mixture was 15 stirred for 16 hr at room température. The solids were filtered out. The fîltrate was diluted with

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

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

[0187] Into a 100-mL round-bottom flask, was placed a solution of methyl 2-[(lE)-2-cyanoethl-en-l-yl]pyridîne-3-carboxyiate (1.4 g, 7.44 mmol, 1 equiv) in MeOH (20 mL), a solution of NaOH (0.6 g, 14.88 mmol, 2 equiv) in H2O (4 mL). After stirring for 2 hr at room température, the reaction was diluted with 10 mL of H2O, pH adjusted to 6-7 with HCl (2 mol/L), and then 25 concentrated. The crude product was purified by Flash-Prep-HPLC with the following conditions (IntelFlash-1): Column, Cl8 silica gel; mobile phase, H₂O:ACN=10:l increasing to H2O:ACN-1:1 with 10 min. This provided the tîtle compound. LCMS (ES) [M+l]⁺ m/z 175.0.

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

[0188] Into a 100-mL round-bottom flask, was placed a solution of 2-[(lE)-2-cyanoeth-l-en-l30 yl]pyridine-3-carboxylic acid (600 mg, 3.45 mmol, 1 equiv) in MeOH (20 mL), palladium on carbon (“Pd/C,” 120 mg, 1.13 mmol, 0.33 equiv). The resulting solution was stirred for 16 hr at room température under H2 atmosphère (20 atm). The solids were filtered out. The filtrate was concentrated to give the crude product. LCMS (ES) [M+l]⁺ m/z 177.1.

Step 5: Synthesis of (S)-3-(3-(2-(hydroxymethyl)pipendine-l-carboiiyl)pyridin-2yl)propanenitrile (3i).

[0189] Into a 50-mL round-boltom flask, was placed a solution of 2-[(lE)-2-cyanoeth-l-en-lylJpyridine-3-carboxylic acid (550 mg, 3.16 mmol, 1 equiv) in DMF (6 mL), [(2S)-piperidin-2yl]methanol (545.6 mg, 4.74 mmol, 1.5 equiv), DIEA (816.3 mg, 6.32 mmol, 2 equiv), HATU (1801.2 mg, 4.74 mmol, 1.5 equiv). The resulting solution was stirred for 4 hr at room température. The mxiture was diluted with 40 mL of H2O and extracted with 3x30 mL of ethyl acetate. The combined organic layers was washed with 30x30 mL of brine, dried over NazSCL, filtered and concentrated. The resulting residue was purified by Flash-Prep-HPLC with the followîng conditions (IntelFlash-1): Column, C18 silica gel; mobile phase, H2O:ACN=1Û:1 increasing to H2O:ACN=1:1 with 10 min. This provided the title compound. LCMS (ES) [M+lfm/z 274.1.

Step 6: Synthesis of(S)-3-(3-(2-((2-formyl-3-hydroxyphenoxy)methyl)piperidine-lcarbonyl)pyridin-2-yl)propanenitrile (Compound 3).

[0190] Into a 50-mL round-bottom flask, was placed a solution of 3-[3-[(2S)-2(hydroxymethyl)piperidine-l-carbonyl]pyridin-2-yl]propanenitrile (200 mg, 0.73 mmol, 1 equiv) in DCM (3 mL), 2,6-dihydroxybenzaldehyde (202.1 mg, 1.46 mmol, 2 equiv), PPh₃ (383.8 mg, 1.46 mmol, 2 equiv). This was followed by the addition of DBAD (337.0 mg, 1.46 mmol, 2 equiv) at 0 ^ÜC. The resulting mixture was stirred for 2 hr at room température and then concentrated under vacuum; the residue was diluted with 5 mL of ACN and filtered. The crude product was purified by Prep-HPLC with the followîng conditions (Prep-HPLC-007): Column, SunFire Prep C18 OBD Column, 150mm 5um lOnm; mobile phase, Water (0.1% formic acid) and MeOH (40% Phase B up to 55% in 7 min, hold 95% in 1 min, down to 40% in 1 min, hold 40% in 1 min); Detector, UV. This provided the title compound. ^lHTEM NMR (300 MHz, 353K, DMSO-d_e):Ô 11.60 (br, 1 H), 10.27 (br, 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 (br, 1H), 4.32 (dd, J= 10.3,6.2 Hz, 1H), 3.21 -2.89 (m, 6H), 1.91-1.46 (m,

6H). LCMS (ES) [M+l]⁺ m/z 394.1.

Example 4: (S)-2-hydroxy-6-((1-(2-(2-(pyrrolidin-l-yl)etliyl)nicotïnoyI)piperidin-2yl)methoxy)benzaldehyde, Compound 4

[0191] Compound 4 was synthesized according to Scheme 4.

Schenie 4

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

[0192] Into a 100-mL round-bottom flask, was placed a mixture of methyl 2-chloropyridine-3 carboxylate (3 g, 17.48 mmol, 1.00 equiv), dioxane (40 mL), water (4 mL), 2-etheny 1-4,4,5,5tetramethyl-l,3,2-dioxaborolane (5.39 g, 34.99 mmol, 2.00 equiv), CsîCOj (11.40 g, 34.99 mmol, 2.00 equiv) and Pd(PPhs)4 (2.02 g, 1.75 mmol, 0.10 equîv). The resulting solution was stirred for 2 h at 100 °C under Ni- The reaction mixture was cooled, filtered, and concentrated under vacuum. The resulting residue was purîfied by a silica gel column by elutîng with ethy 1 acetate/petroleum ether (1/2) to give 2 methyl 2-ethenylpyridine-3-carboxylate. LCMS (ES) [M+l ] ⁺ m/z: 164.1.

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

[0193] Info a 100-mL round-bottom flask, was placed a solution of methyl 2-ethenylpyridine5 3-carboxylate (3.0 g, 18.39 mmol, 1 equiv), MeOH (50 mL), H2O (5 mL) and NaOH (3.7 g, 91.93 mmol, 5.0 equiv). After stirring for 2 h at 50°C, the reaction mixture was cooled, and pH was adjusted to 5 with addition of aqueous HCl (2 M). The resulting mixture was concentrated and diluted with 100 mL of DCM. The solids were filtered out. The mixture was concentrated. This resulted in 2-ethenylpyridine-3-carboxylic acid. LCMS (ES) [M+l] ⁺ m/z: 150.1.

Step 3: Synthesis of tert-butyl (2S)-2-[[(2,2-dimethyl-4-oxo-2,4-dihydro-l,3-benzodioxin-5yl)oxy] methyl] pi peridine-l-carboxylate (4e).

[0194] Compound 4d may be synthesized according to methods known in the art.

[0195] Into a 1000-mL round-bottom flask, was placed a solution of 5-hydroxy-2,2-dimethyl2,4-dihydro-l,3-benzodioxin-4-one (4d, 10.0 g, 51.50 mmol, 1 equiv), THF (300 mL), tert-butyl (2S)-2-(hydroxymethyl)piperidine-l-carboxylate (22.2 g, 103.12 mmol, 2.00 equiv) and PPI13 (40.5 g, 154.49 mmol, 3 equiv). It was added the solution of diisopropyl azodicarboxylate (“DIAD,” 31.2 g, 154.49 mmol, 3 equiv) in THF (30 ml) dropwise at 0°C under N2. The resulting solution was stirred for 4 h at room température. The resulting mixture was concentrated. The residue was purified by a silica gel column by eluting with ethyl acetate/petroleum ether (1/3). This resulted in tert-butyl (2S)-2-[[(2,2-dimethyl-4-oxo-2,4dihydro-l,3-benzodioxin-5-yl)oxy]methyl]piperidine-l-carboxylate. LCMS (ES) [M+l] ⁺ m/z: 392.2.

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

[0196] Into a 50-mL round-bottom flask, was placed a solution of tert-butyl (2S)-2-[[(2,2dimethyl-4-oxo-2,4-dihydro-l,3-benzodioxîn-5-yl)oxy]methyl]piperidine-l-carboxylate (2.0 g, 5.10 mmol, 1 equiv), DCM (15 mL) and HCl/dioxane (4 M, 5 mL). The resulting solution was stirred for 1 h at room température. The resulting mixture was concentrated. This resulted in 2,2dimethyl-5-[[(2S)-piperidin-2-yl]methoxy]-2,4-dihydro-l,3-benzodioxin-4-one. LCMS (ES)

[M+l] ⁺ m/z: 292.2.

Step 5: Synthesis of 5-[[(2S)-l-(2-ethenyIpyridine-3-carbonyl)piperïdïn-2-yl]methoxy]-2,2dimethyl^^-dihydro-ljS-benzodioxin^-one (4g).

[0197] Into a 100-mL round-bottom flask, was placed a solution of 2,2-dimethyl-5-[[(2S)piperidin-2-yl]methoxy]-2,4-dihydro-l,3-benzodioxin-4-one hydrochloride (1.0 g, 3.05 mmol, 1 5 equiv), DCM (50 mL, 786.50 mmol, 257.82 equiv), 2-ethenylpyridine-3-carboxylic acid (910.0 mg, 6.10 mmol, 2.00 equiv), DIEA (2.0 g, 15.25 mmol, 5 equiv) and HATU (2.3 g, 6.10 mmol, 2 equiv) at 0 °C. After stirring 2 h at room température, the reaction mixture was diluted with 50 mL of DCM and washed with 3x50 ml of brine. The organic layer was drîed over anhydrous sodium sulfate and concentrated. The residue was purified by a silica gel column by eluting with ethyl acetate/petroleum ether (1/1). This resulted in 5-[[(2S)-l-(2-ethenylpyridine-3carbonyl)piperidin-2-yl]methoxy]-2,2-dimethyl-2,4-dihydro-l,3-benzodioxin-4-one. LCMS (ES) [M+l]⁺ m/z: 423.2.

Step 6: Synthesis of2,2-dimethyl-5-[[(2S)-l-[2-[2-(pyrrolidin-l-yl)ethyl]pyridine-3carbonyl]piperidin-2-yl]methoxy]-2,4-dihydro-l,3-benzodioxin-4-one (4h),

[0198] Into a 50-mL round-bottom flask, was placed a solution of 5-[[(2S)-l-(2ethenylpyridine-3-carbonyl)piperidîn-2-yl]methoxy]-2,2-dimethyl-2,4-dihydro-l ,3-benzodioxin4-one (650 mg, 1.54 mmol, 1 equiv), éthanol (20 mL), pyrrolidine (218.8 mg, 3.08 mmol, 2.00 equiv) and TEA (311.4 mg, 3.08 mmol, 2 equiv). The réaction mixture was stirred for 16 h at 85°C, cooled and concentrated in vacuum. The resulting residue was purified by a silica gel column by eluting with dichloromethane/methanol (10/1 ). This resulted in 2,2-dimethyl-5[[(2S)-l-[2-[2-(pynOÎidin-!-yl)ethyl]pyridine-3-carbonyl]piperidin-2-yl]methoxy]-2,4-dihydro1,3-benzodioxîn-4-one. LCMS (ES) [M+l]⁺ m/z: 494.3.

Step 7: Synthesis of2-(hydroxymethyl)-3-[[(2S)-l-[2-[2-(pyrrolidin-l-yl)ethyl]pyridine-3carbonyl]piperidin-2-yl]methoxy] phénol (4i).

[0199] Into a 50-mL 3-necked round-bottom flask, was placed a solution of 2,2-dimethyl-5[[(2S)-l-[2-[2-(pyrrûlidin-l-yl)ethyI]pyridine-3-carbonyl]piperidin-2-yl]methoxy]-2,4-dihydrûl,3-benzodioxin-4-one (500 mg, 1.01 mmol, 1 equiv) and THF (10 mL). To this was added lithium aluminum hydride in THF solution (“LiAlH^i THF solution,” 2.03 mL, 1 M, 2.03 mmol, 2 equiv) dropwise at -78 C under N2. The resulting mixture was stirred at -78 °C for 1 h. The reaction mixture was warmed to 0 °C, and then to this was added, dropwise, 0.07 mL of H2O, 0.07 mL of 15% aqueous NaOH and 0.21 mL of H2O. The mixture was warmed to room température and stirred for 30 minutes. The solids were filtered out. The filtrate was concentrated. This resulted in 2-(hydroxymethyl)-3-[[(2S)-l-[2-[2-(pyrrolidin-lyl)ethyl]pyridine-3-carbonyl]piperidin-2-yl]methoxy]phenol. LCMS (ES) [M+l] * m/z: 440.3.

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

[0200] Into a 50-mL round-bottom flask, was placed a mixture of 2-(hydroxymethyl)-3-[[(2S)l-[2-[2-(pyrrolidin-l-yl)ethyl]pyridme-3-carbonyl]piperidin-2-yl]methoxy]phenol (200 mg, 0.46 mmol, 1 equiv), DCM (10 mL) and MnCL (791.1 mg, 9.10 mmol, 20.00 equiv). The resulting mixture was stirred for 1 h at room température. The reaction mixture was fîltered and concentrated. The resulting residue was purified by reverse phase préparative HPLC (Prep-C18, 10 5 mM XBridge column, 19 x 150 mm, waters; gradient elution of 15% MeCN in water to 35%

MeCN in water over a 6 min perîod, where both solvents contaîn 0.1% TFA) to provide 2hydroxy-6-[[(2S)-l-[2-[2-(pyrrolidm-l-yl)ethyl]pyridine-3-carbonyl]piperidin-2yl]methoxy]benzaldehyde. ’H NMR (300 MHz, DMSO-dô) δ 11.57 (s, IH), 10.26 (s, 1 H), 9.48 (s, 1H), 8.60 (dd,7 = 4.8, 1.8 Hz, 1H), 7.65 (s, 1H), 7.53 (t,7 = 8.3 Hz, 1H), 7.38 (dd, 7 = 7.7, 15 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, 7 = 7.2 Hz, 2H), 3.51 - 2.96 (m, 8H), 2.07-1.84 (m, 5H), 1.81-1.36(m, 5H). LCMS (ES) [M+l] ⁺ m/z: 438.2.

Example 5: (S)-2-hydroxy-6-((l-(2-(hydroxymethyl)benzoyl)piperidin-2yl)methoxy)benzaldehyde, Compound 5

[0201] Compound 5 was synthesized according to Scheme 5.

Step 1

NaOH.HjO

5a 5b

Scheme 5

[0202] In Scheme 5, TBDMSC1 refers to tert-butyldimethylsilyl chloride, and MsCI refers to mesyl chloride. Compound 5: MS m/z 370.2 [M+H]⁺, 392.2 [M+Na]⁺.

Example 6: (R)-2-hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoy!)thioinorpholin-3yl)methoxy)benzaldehyde and (S) -2-hydroxy-6-((4-(2-(2hydroxyethyl)nicotinoyl)thioinorpholin-3-yl)inethoxy)benzaldehyde

Scheme 6A

Enantiomère 1 and 2 of 6b

Steps 4a & 4b

Compound 10 Enantiomers 1 and 2

Step 1: Synthesis of [4-(2-[2-[(tert-butyldimethylsilyl)oxylethyl]pyridine-3carbonyl)thiomorpholin -3 -yI] methanoi (6a).

[0203J Into a 100-mL 3-neckcd round-bottom flask, was addded 2-[2-[(tertbulyldimethylsilyl)oxy]ethyl]pyridine-3-carboxylîc acid (2.00 g, 7.11 mmol, 1.00 equiv), thiomorpholm-3-ylmethanol (0.95 g, 7.13 mmol, 1.00 equiv), DIEA (2.76 g, 21.32 mmol, 3.00 equiv) and DCM (30.00 mL). To this mixture was added HATU (3.24 g, 8.53 mmol, 1.20 equiv), in portions at 0 °C. The resulting reaction mixture was allowed to warm to room température and stirred for overnight. The reaction was then quenched by the addition of 30 mL of water. The resulting solution was extracted with 3x30 mL of dichloromethane and the organic layers were separated, combined and dned over anhydrous sodium sulfate and concentrated. The residue was applied onto a silica gel column with THF/petroleum ether (“PE”) (30%) as eluent. The combined fractions were concentrated to produce [4-(2-[2-[(tertbutyldimethylsiIyl)oxy]ethyl]pyridine-3-carbonyl)thiomorpholin-3-yI]methanol. LCMS (ES) [M+H]+m/z: 397.

Step 2. Synthesis of2-[[4-(2-[2-[(tert-butyldÎmethylsilyl)oxy]ethyl]pyrÎdîne-3carbonyI)thïomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde (6b).

[0204] Into a 100-mL 3-necked round-bottom flask, was added [4-(2-[2-[(tertbutyldimethylsilyl)oxy]ethyl]pyridine-3-carbonyl)thiomorpholin-3-yl]methanol (1.50 g, 3.78 mmol, 1.00 equiv), 2,6-dihydroxybenzaldehyde (0.63 g, 4.56 mmol, 1.21 equiv), PPh3 (1.19 g, 4.54 mmol, 1.20 equiv), and DCM (30.00 mL). To this solution was added DIAD (0.92 g, 4.54 mmol, 1.20 equiv) dropwise over 20 mins with stirring at 0 °C. The resulting mixture was stirred overnight at room température, and was concentrated. The residue was directly applied onto a silica gel column with THF/PE (25%) as eluent. The combined fractions were concentrated to give 2-[[4-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridine-3-carbonyI)thioinorpholin-3yl]methoxy]-6-hydroxybenzaldehyde. LCMS (ES) [M+H]+ m/z:517.

Step 3. Chîral-HPLC séparation of Compound 6b.

[0205] The racemate was purified by Chiral-HPLC to give Enantiomer 1 and Enantiomer 2 of Compound 6b with the following conditions: Column, Lux Cellulose-4, 4.6*100 mm, 3 pm; mobile phase, A: n-Hexane B: Ethanol (35% B in 18 min); Flow rate: 30 mL/min; Detector, 254. LCMS (ES) [M+H]+ m/z:517 (for both compounds).

Step 4a. Reinoval of TBS group to give Compound 10, Enantiomer 1.

[0206] HCl (~2M) in 5 ml of ethyl acetate (“EA”) was added to Enantiomer 1 of Compound 6b (335.00 mg, 0.65 mmol, 1.00 equiv) ΐη EA (3.00 mL) dropwise with stirring at 0 °C. The resulting solution was stirred for 2 h al room température. The pH value of the solution was adjusted to 8 with saturated NaHCCh. The resulting solution was extracted with 3x10 mL of ethyl acetate and the organic layers combined, drïed over anhydrous sodium sulfate, filtered and concentrated. The cru de product was purified by Prep-HPLC with the following conditions: Column, XBridge Prep C18 OBD Column, 19cm, 150mm, 5um; mobile phase, Water (0.1% HCOOH) and CAN (30% Phase B up to 40% in 10min); Detector, 254. This resulted in

Enantiomer 1 of Compound 10 with rétention time = 4.06 min. LCMS (ES) [M+H]⁺m/z: 403.1;

[M+Na]⁺m/z: 425.1.

Step 4b. Removal of TBS group to give Compound 10, Enantiomer 2.

[0207] HCl (~2M) in 5 ml of EA was added to Enantiomer 2 of Compound 6b (335.00 mg, 5 0.65 mmol, 1.00 equiv) in EA (3.00 mL) dropwise with stirring at 0 ^UC. The resulting solution was stirred for 2 h at room température. The pH value of the solution was adjusted to 8 with saturated NaHCOa. The resulting solution was extracted with 3x10 mL of ethyl acetate and the organic layers combined, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product (200 mg) was purified by Prep-HPLC with the following conditions: Column, 10 XBridge Prep C18 OBD Column, 19cm, 150mm, 5um; mobile phase, Water (0.1% HCOOH) and CAN (30% Phase B up to 40% in 10 min); Detector, 254. This resulted in Enantiomer 2 of

Compound 10 with rétention time - 5.40 min. LCMS (ES) [M+H]⁺Wz: 403.2; [M+Na]⁺m/z: 425.1.

Alternative Synthesis of (R)-2-hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl)thiomorpholin15 3-yl)methoxy)benzaIdehyde

[0208] (R)-2-hydroxy-6-((4-(2-(2 -hydroxyethy l)nicotinoy l)thiomorphoIin-3 yl)methoxy)benzaldehyde can be made directly from chiral (R)-thiomorpholin-3-ylmethanol as depicted in Scheme 6B.

Scheme 6B

Step 1

[0209] To a solution of L-cysteine (100.0 g, 825.4 mmol, 1.0 equiv) in H₂O (1.0 L) was added

NaOH (3.3 g, 82.5 mmol, 0.1 equiv). After the reaction mixture was cooled to 0 °C, ethylene oxide (100.0 g, 2.26 moi, 2.75 equiv) was added dropwise. The resulting solution was stirred for 3 hr at 0-25 °C in a water/ice bath. The resulting solution was extracted with 3x500 mL of ethyl acetate to remove unchanged ethylene oxide. The aqueous layer was concentrated under reduced pressure. The crude product was trîturated with EtOH (200 mL) for Ih and filtered. This resulted in hy droxy ethylcysteine. LCMS (ES) [M+l] ⁺ m/z: 166.2; Rétention time 0.174 min. ’H-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).

Step 2

[0210] To a mixture of hydroxyethylcysteine (130.0 g, 786.8 mmol, 1.0 equiv) and KHCO3 15 (165.4 g, 1.65 mol, 2.1 equiv) in dioxane (700 mL, 8.26 mol, 10.5 equiv) and H₂O (700 mL) was added CbzCl (147.6 g, 865 mmol, 1.1 equiv) dropwise at 0 °C over 30 min. The resulting solution was stirred for 5 h at 0-25 °C. The solvents evaporated off and the residue dissolved in DMF (1000 mL). BnBr (148 g, 0.86 mol, 1.1 equiv) was added and the resulting mixture was stirred for 16 h at 0-25 °C. The reaction was then quenched by the addition of 1000 mL of water. The resulting solution was extracted with 3x1000 mL of EtOAc, the combined organic 5 layer was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:50 to 1:5). This resulted in benzyl (2R)-2-[[(benzyloxy)carbonyl]amino]-3-[(2hydroxyethyl)sulfanyl]propanoate. LCMS (ES) [M+l] ⁺ m/z: 390.5; Rétention time 1.146 min, ‘H-NMR: (300 MHz, CDCh, ppm): δ 7.39-7.33 (m, 10H), 5.83 (br, 1H), 5.26 (d, J = 4.7 Hz, 10 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).

Step 3

[0211] Into a 2500-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of argon, was placed benzyl (2R)-2-[[(benzyloxy)carbonyl]amîno]-3-[(2hydroxyethyl)sulfanyl]propanoate (90.0 g, 231 mmol, 1.0 equiv), THF (1.0 L), DEAD (48.3 g, 15 277 mmol, 1.2 equiv). After the reaction was cooled to 0 ⁽’C, PPhs (78.8 g, 300 mmol, 1.3 equiv) in THF (100 mL) was added dropwise. The resulting solution was stirred for 16 h at 0-25 °C.

The resulting mixture was concentrated. The residue was applied onto a silica gel column with THF/PE (1:100 to 1:5). This resulted in 3,4-dibenzyl (3R)-thiomorpholine-3,4-dicarboxylate. LCMS (ES) [M+l] ⁺ m/z: 372.1; Rétention time 1.312 min.

Step 4

[0212] Into a 2500-mL 3-necked round-bottom flask, was placed 3,4-dibenzyl (3R)thiomorpholine-3,4-dicarboxylate (100.0 g, 269 mmol, 1.0 equiv), DCM (1.0 L). The reaction was cooled to 0 ⁽’C, TMSI (161.6 g, 0.81 mol, 3 equiv) was added dropwise. The resulting solution was stirred for 1 h at 0-25 °C in a water/ice bath. The reaction was then quenched by the 25 addition of 100 mL of MeOH. The resulting mixture was concentrated. The pH value of the solution was adjusted to 1 with HCl (2 mol/L). The resulting solution was extracted with 2x500 mL of MTBE, and the aqueous layers were combined. NaHCOs (2 mol/L) was employed to adjust the pH to 8. The resulting solution was extracted with 3x500 mL of ethyl acetate. The combined organic layer was dried over Na₂SO₄ and concentrated under reduced pressure. This resulted in benzyl (3R)-thiomorpholine-3-carboxylate. LCMS (ES) [M+l] ⁺ m/z: 238.1;

Rétention lime 1.026 min; 'H-NMR: (300 MHz, CDCh, ppm): δ 7.43-7.33 (m, 5H), 5.25 (s, 2H), 3.74 (dd, 7 = 8.6, 3.4 Hz, 1H),3.4O (ddd, J = 12.5, 4.9, 3.0 Hz, 1 H), 3.04 (ddd,7 = 12.5,

9.8, 2.7 Hz, 1H), 2.90 (ddd,7= 13.2, 3.4,1.3 Hz, 1H), 2.82 (dd, J - 13.3, 8.6 Hz, 1H), 2.70 (ddd,J= 12.9,9.8,3.0 Hz, 1H), 2.48 (dddd, J = 13.3, 4.9, 2.7, 1.3 Hz, 1H).

Step 5

[0213] To a suspension of L1AIH4 (13.2 g, 347 mmol, 1.5 equiv) in THF (1000 mL) was added 5 benzyl (3R)-thiomorpholine-3-carboxylate (55.0 g, 231.7 mmol, 1.0 equiv) in THF (100 mL) drop wise at 0 °C. After the resulting solution was stirred for 3 hr at 0-25 °C, the reaction was then quenched by the addition of 100 g of NazSOr I0H2O. The resulting solution was diluted with 500 mL of THF, and the solids were filtered out. The resulting mixture was concentrated, and the residue was applied onto a silica gel column with THF/PE (1:50 to 2:1). This resulted in 10 (3R)-thiomorpholin-3-ylmethanol. LCMS (ES) [M+l] ⁺ m/z: 134.1; Rétention time 0.464 min;

‘H-NMR: (300 MHz, DMSO-t/6, ppm): δ 4.64 (br, IH), 3.26-3.17 (m, 2H), 2.81-2.68 (m, 4H), 2.43-2.26 (m, 4H).

Step 6

[0214] Into a 300-mL 3-necked round-bottom flask, was placed 2-[2-[(tert15 butyldimethylsiIyl)oxy]ethyl]pyridine-3-carboxylic acid (20.00 g, 71.06 mmol, 1.00 equiv), (3R)-thiomorphoiin-3-ylmethanol (10.41 g, 78.14 mmol, 1.10 equiv), DCM (300.00 mL), and DIEA (18.37 g, 142.13 mmol, 2.00 equiv). This was followed by the addition of HATU (32.43 g, 85.28 mmol, 1.20 equiv), in portions at 0 °C. The resulting solution was stirred for 3 h at room température. The reaction was then quenched by the addition of 200 mL of water. The resulting solution was extracted with 3x200 mL of dichloromethane, and the organic layers were combined and dried over anhydrous sodium sulfate and concentrated. The residue was applied onto a silica gel column with THF/PE (30%). This resulted in [(3R)-4-(2-[2-[(tertbutyldimethylsilyl)oxy]ethyl]pyridine-3-carbonyl)thiomorpholin-3-yl]methanol. LCMS (ES) [M+H]⁺ m/z: 397.30.

Step 7

[0215] Into a 1000-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed 2,6-dihydroxybenzaldehyde (7.52 g, 54.45 mmol, 1.20 equiv), [(3R)-4-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridine-3-carbonyl)thiomorphoHn-3yl]methanol (18.00 g, 45.38 mmol, 1.00 equiv), PPlv (14.28 g, 54.46 mmol, 1.20 equiv), and 30 DCM (400.00 mL). This was followed by the addition of DIAD (11.01 g, 54.46 mmol, 1.20 equiv) dropwise with stirring at 0 °C. The resulting solution was stirred for overnight at room température. The resulting mixture was concentrated. The residue was applied onto a silica gel column with THF/PE (15%). This resulted in 2-[[(3R)-4-(2-[2-[(tertbutyldimethylsilyl)oxy]ethyl]pyridine-3-carbony!)thîomorpholîn-3-yI]methoxy]-6hydroxybenzaldehyde. LCMS (ES) [M+H]⁺ m/z: 517.35.

Step 8

[0216] Into a 500-mL round-bottom flask, was placed 2-[ [(3R)-4-(2-[2-[(tertbutyidimethylsilyl)oxy]ethyl]pyridine-3-carbonyl)thiomorpholin-3-yl]methoxy]-6hydroxybenzaldehyde (13.50 g, 26.13 mmol, 1.00 equiv) and EA (20.00 mL). To the above HCl(g) in EA (52.25 mL, 104.50 mmol, 4.00 equiv) was introduced in dropwise with stirring at 10 0 °C. The resulting solution was stirred for 2 h at room température. The reaction was then quenched by the addition of 80 mL of water. The pH value of the solution was adjusted to 7-8 with saturated NazCQj. The resulting solution was extracted with 3x100 mL of dichloromethane, and the organic layers combined and dried in an oven under reduced pressure, and concentrated.

The crude product was purified by Prep-HPLC with the following conditions: Column, XBridge 15 Prep C18 OBD Column, 19cm, 150mm, 5um; mobile phase, Water (0.1% HCOOH) and ACN (30% Phase B up to 50% in 11 min); Detector, 254. This resulted in 2-hydroxy-6-[[(3R)-4-[2-(2hydroxyethyl)pyridine-3-carbonyl]thiomorpholîn-3-yl]methoxy]benzaldehyde.

[0217] Chiral HPLC conditions were as follows: Instrument: SHIMADZU LC-20AT; Mobile

Phase A: n-Hexane(0.1%TFA); Mobile Phase B: Ethanol; Conc. of Phase B: 50.0%; Flow Rate: 20 1.000 mL/min; Column: Lux Cellulose-4, 4.6*100 mm, 3pm. Chiral HPLC rétention time = 5.41 min.

[0218] LCMS (ES, m/z)·. [M+H] ⁺: 403.2; NMR (300 MHz, DMSO-d6): δ 11.80-11.73 (m, 1H), 10.33 (br, 1H), 8.56 (dd, J = 4.9, 1.8 Hz, 1H), 7.90-7.39 (m, 2H), 7.37-7.19 (m, 1H), 6.816.63 (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), 25 3.20-2.61 (m, 6H), 2.43 (d, J = 12.6 Hz, 1H).

[0219] Based on the product of Scheme 6B, it was determined that Compound 10, Enantiomer 2 corresponds to (Æ)-2-hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl)thiomorpholin-3yl)methoxy)benzaldehyde.

Example 7: ( S)-2-hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl) morpholin-3 30 yl)methoxy)benzaldehyde, Compound 8

[0220] Compound 8 was synthesized according to Scheme 7.

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

[0221] To a solution of 2-[2-[(tert-butyIdimethylsilyl)oxy]ethyi]pyridine-3-carboxyIic acid (1.50 g, 5.33 mmol, 1.00 equiv) and (3R)-morpholin-3-ylmethanol hydrochloride (0.98 g, 6.39 mmol, 1.20 equiv) in DCM (20 mL) was added DIEA (2.07 g, 15.99 mmol, 3.00 equiv), foilowed by the addition of HATU (2.43 g, 6.39 mmol, 1.20 equiv) in portions over 5 mins. The resulting solution was stirred for 2 hr at room température, diluled with 50 mL of H2O. The resulting solution was extracted with 2x30 mL of dichloromethane and the organic layers were separated, combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to gîve a residue that was purified ou silica gel column with ethyl acetate/petroleum ether (2/1) as eluent. This resulted in (R)-(2-(2-((tertbutyldimethylsilyl)oxy)ethyl)pyridin-3-yl)(3-(hydroxymethyl) morpholino)methanone. LCMS 15 (ES) [M+H]Wz: 381.2.

* Step 2, Synthesis of (S)-2-((4-(2-(2-((tertbutyldiniethylsilyl)oxy)ethyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxy benzaldehyde (7b).

[0222] A solution of (R)-(2-(2-((tert-butyldimethylsilyl)oxy)ethyl)pyridin-3-yl)(35 (hydroxymethyl) morpholino)methanone (600 mg, 1.57 mmol, 1.00 equiv), 2,6dihydroxybenzaldehyde (261 mg, 1.89 mmol, 1.20 equiv), and PPh3 (496 mg, 1.89 mmol, 1.20 equiv) in DCM (10 mL) was purged and maintained with an inert atmosphère of nitrogen. To tins mixture was added DIAD (382 mg, 1.89 mmol, 1.20 equiv) dropwise with stirring at 0°C over 5 min. The resulting solution was stirred for 1 hr at room température, diluted with 20 mL of H2O. The resulting solution was extracted with 2x20 mL of dichloromethane and the organîc layers combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give the crude product which was purified by Flash-Prep-HPLC with the followîng conditions (IntelFlash-1): Column, C18 silica gel; mobile phase, H2Ü(0.1% HCOOH)/acetonitrile (“ACN”)=2/1 increasing to H2O (0.1% HCOOH)/ACN-l/4 within 18 min; Detector, UV 254 nm. This resulted in (S)-2-((4-(2-(2-((tertbutyldimethylsilyl)oxy)ethyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybenzaldehyde. 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 (8).

[0223] Formic acid (HCOOH, 1 ml) was added to a solution of (S)-2-((4-(2-(2-((tertbutyldimethylsilyl)oxy)ethyl)nicotînoyl)morpholin-3-yl)methoxy)-6-hydroxy benzaldehyde (450 mg, 0.89 mmol, 1.00 equiv) in ACN (5.00 mL). The resulting solution was stirred for 3 hr at 40 ^ÜC, cooled room température and diluted with 5 mL of ACN. The mixture was concentrated to give the crude product, which was purified by Prep-HPLC with the following conditions (2#SH1MADZU (HPLC-01)): Column, Atiantis HILIC OBD Column, 19^:h150mm*5um; mobile phase, Water(0.1% FA) and ACN (37% PhaseB up to 45% in 10 min); Detector, UV 254 nm. This resulted in (S)-2-hydroxy-6-((4-(2-(2-hydroxyethyl)nicotmoyl)morpholîn-3yl)methoxy) benzaldehyde. LCMS (ES) [M+H]⁺/m/z: 387.1.

Example S. (S)-2-hydroxy-6-((1-(2-(hydroxy methyl) nicotinoyl) piperidin-2yl)methoxy)benzaldehyde, Compound 11

[0224] Compound 11 was synlhesized according to Scheme 8.

Scheme 8

8d 8e

8f

Step 8

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

[0225] To a solution of methyl 2-methylpyridine-3-carboxylate (15.00 g, 99.23 mmol, 1.0 equiv) in DCM (150 mL) at 0°C was added 3-chIoroperbenzoic acid (“m-CPBA,” 34.4 g, 199.34 mmol, 2.0 equiv). The reaction mixture was stirred for 2 h at room température. The reaction was quenched with saturated aqueous Na₂CO3 (100 mL), and the organic phase was separated out and drîed over anhydrous sodium sulfate. The solution was filtered, and the fîltrate was concentrated under reduced pressure. The residue was purified by silica gel column with 5 dichloromethane/methanol (10/1), providing 3-(methoxycarbonyl)-2-methylpyridine 1-oxide.

LCMS (ES) [M+Hf m/z: 168.

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

[0226] A mixture of 3-(methoxycarbonyl)-2-methylpyridme 1-oxide (8.00 g) in acetic anhydride (80 mL) was heated for 5 h at 140 °C. After being cooled to room température, excess 10 liquid was removed under reduced pressure, and the residue was suspended in water (50 mL) and extracted with 3x50 mL of dichloromethane. The combined organic phase was dried over anhydrous sodium sulfate and filtered, and the fîltrate was concentrated under reduced pressure. The residue was purified by silica gel column with ethyl acetate/petroleum ether (15%), providing methyl 2-(acetoxymethyl)nicotinate. LCMS (ES) [M+H]⁺nz/z: 210.

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

[0227] To a solution of methyl 2-(acetoxymethy])nicotinate (7.90 g, 37.76 mmol, 1.0 equiv), in MeOH (80 mL) was added acetyl chloride (3.60 g, 45.86 mmol, 1.2 equiv). The réaction solution was stirred overnight at room température; 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 NaHCO₃ solid and extracted with ethyl acetate (30 mL*3). The combined organic phase was dried over anhydrous sodium sulfate and filtered, and the fîltrate was concentrated in vacuum. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/1), giving methyl 2-(hydroxymethyl)pyridine-3-carboxylate. LCMS (ES) [M+l]⁺ mlz: 168.

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

[0228] Into a 100-raL 3-necked round-bottom flask, was placed methyl 2(hydroxymethyl)pyridine-3-carboxylate (2.80 g, 16.75 mmol, 1.0 equiv), DCM (40 mL), and imidazole (2.27 g, 33.34 mmol, 2.0 equiv). This was followed by the addition of tbutyldimethylchlorosilane (4.04 g, 26.81 mmol, 1.6 equiv) at 0 °C. The mixture was stirred for 2 30 h at room température. The reaction was then quenched by the addition of water (30 mL), extracted with 3x50 mL of dichloromethane. The combined organic phase was dried over anhydrous sulfate and filtered, and the filtrate was concentrated in vacuum. The residue was purified by silica gel column with ethyl acetate/petroleum ether (10%), giving methyl 2-(((tertbutyldimethylsilyl)oxy)methyl)nicotinate. LCMS (ES) [M+l]⁺ m/z: 282.

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

[0229] Into a 100-mL 3-necked round-bottom flask, was placed methyl 2-(((tertbutyldimethylsilyl)oxy)methyl)nicotinate (4.20 g, 14.92 mmol, 1.0 equiv), MeOH (30 mL), and H2O (15 mL). This was followed by the addition of LiOH H2O (1.25 g, 29.79 mmol, 2.0 equiv) at 0°C. The mixture was stirred for 2 h at room température, then concentrated to remove the solvent, and the pH value of the residue was adjusted to 7 with citric acid. The solution was filtered, and the solid was dried under infrared lamp. 2-(((tertbutyldimethylsilyl)oxy)methyl)nîcotinic acid was obtained. LCMS (ES) [M+l]* m/z: 268.

Step 6. Synthesis of (S)-(2-(((tert-butyldimethyIsilyl)oxy)methyl)pyridin-3-yl)(2(hydroxymethyl)piperidin-l-yl)inethanone

[0230] Into a 50-mL 3-necked round-bottom flask, was placed 2-(((tert15 butyldimethylsilyl)oxy)methyl)nicotinic acid (615 mg, 2.30 mmol, 1.0 equiv), (2S)-piperidin-2ylmethanol (318 mg, 2.76 mmol, 1.2 equiv), DCM (10 mL), DIEA (594 mg, 4.60 mmol, 2.0 equiv). This was followed by the addition of HATU (1.05 g, 2.76 mmol, 1.2 equiv) at 0 °C. The mixture was stirred for 2 h at room température. The reaction mixture was concentrated to remove solvent, and the residue was purified by silica gel column with ethyl acetate/petroleum ether (80%). (S)-(2-(((tert-butyldimethylsilyl)oxy)methyl)pyridin-3-yl)(2(hydroxymethyi)pîperidin-l-yl)methanone was obtained. LCMS (ES) [M+l]* m/z: 365.

Step 7. Synthesis of (S)-2-((l-(2-(((tert-butyldimethylsilyl)oxy)methyl)nicotinoyI)piperidiii2-y I) methoxy)-6-hyd roxy benzaldehy de

[0231] Into a 40-mL vial purged and maintaîned with an inert atmosphère of nitrogen, was placed (S)-(2~(((tert-butyidimethylsilyl)oxy)methyl)pyridin-3-yl)(2-(hydroxymethyl)piperidin-lyl)methanone (316 mg, 0.87 mmol, 1.0 equiv), 2,6-dihy droxy benzaldehy de (143 mg, 1.04 mmol, 1.2 equiv), PPhj (340 mg, 1.30 mmol, 1.5 equiv), and THF (15 mL). This was followed by the addition of DIAD (262 mg, 1.30 mmol, 1.5 equiv) at 0 °C. After addition, the reaction solution was stirred overnight at room température and then concentrated to remove solvent. The resulting residue was purified by silica gel column with ethyl acetate/petroleum ether (1/1). (S)20663

I

2-((1-(2-(((tert-butykiimethylsilyl)oxy)methyl)nicotmoyI)pjperidm-2-yl)methoxy)-6hydroxybenzaldehyde was obtained. LCMS (ES) [M+l]⁺ m/z: 485.

Step 8. Synthesis of (S)-2-hydiOxy-6-((l-(2-(hydroxymethyl)nicotinoyI)piperidin-2yl)methoxy) benzaldéhyde

[0232] Into a 50-mL round-bottom flask, was placed (S)-2-((l-(2-(((tertbutyldimethylsilyl)oxy)methyl)nicotinoyl)piperidin-2-yl)methoxy)-6-hydroxybenzaldehyde (250 mg, 0.52 mmol, 1.0 equiv) in EA (3ml). To the above solution was added HCl (g) (2 M in EA) (5.0 mL) was added at 0 °C, the mixture was allowed to stir for 1 h at room température. The réaction was then diluted by the addition of water (20 mL), and the pH value of the solution was adjusted to 8 with NaHCO? solid and extracted with 3x20 mL of ethyl acetate. The combined organic phase was concentrated under reduced pressure, and the residue was purified by PrepHPLC with conditions: (2#SHIMADZU (HPLC-01)): Column, Kinetex EVO C18 Column, 21.2*150, 5um, mobile phase, Water (0.1% Formic Acid) and CH₃CN (10% Phase B up to 90% within 15 min), detector, UV 254 nm. (S)-2-hydroxy-6-((l-(215 (hydroxymethyl)nicotinoyl)piperidin-2-yl)methoxy)benzaldehyde was obtained. LCMS-PH(ES, mU}·. [M+H]*; 371.1; [M+Na]⁺: 393.1.

I

Example 9. (S)-2-hydroxy-6-((4-(2-(hydroxymethyl)nicotinoyl)morpholin-3yl)methoxy)bcnzaldehyde, Compound 12

[0233] Compound 12 was synthesized according to Scheme 9 A.

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

[0234] To a solution of 2-[[(iert-butyldimethyIsilyI)oxy]methyl]pyridine-3-carboxylic acid (530 mg, 1.98 mmol, 1.0 equiv), (3R)-morpholin-3-ylmethanol hydrochloride (364 mg, 2.38 mmol, 1.2 equiv), and DIEA (768 mg, 5.94 mmol, 3.0 equiv) in DCM (10 mL) was added HATU (905 mg, 2.38 mmol, 1.2 equiv) at 0°C. The reaction solution was stirred for 3 h at room température. The solution was then concentrated to remove the solvent, and the residue was purified by silica gel column with ethyl acetate/petroleum ether (60%). (R)-(2-(((tertbutyldimelhylsilyl)oxy)methyl)pyridin-3-yl)(3-(hydroxymethyl)morpholino)methanone was obtained. LCMS (ES) [M+l]⁺ m/z: 367.

Step 2. Synthesis of (S)-2-((4-(2-(((tertbutyldimethylsilyl)oxy)methyl)nicotinoyl)morphoIin-3-yl)methoxy)-6hydroxybenzaldehyde

[0235] Into a 40-mL vial purged and maintained with an inert atmosphère of nitrogen, was 5 placed (R)-(2-(((tert-butyldimethyIsilyl)oxy)melhyl)pyridin-3-yl)(3(hydroxymethyl)morpholino)methanone (630 mg, 1.72 mmol, 1.0 equiv), 2,6dihydroxybenzaldehyde (284 mg, 2.06 mmol, 1.2 equiv), PPh^ (540 mg, 2.06 mmol, 1.2 equiv), THF (20 mL). This was followed by the addition of DBAD (474 mg, 2.06 mmol, 1.2 equiv) at 0°C. The reaction solution was stirred overnîght at room température. The solution was concentrated in vacuum to remove die solvent, and the residue was purified by silica gel column with ethyl acetate/petroleum ether (1/1). (S)-2-((4-(2-(((tertbutyldimethylsilyl)oxy)methyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybcnzaldehyde was obtained. LCMS (ES) [M+l]⁺ m/z: 487.

Step 3. Synthesis of (S)-2-hydroxy-6-((4-(2-(hydroxymethyl)nicotînoyl)morpholin-315 yl)niethoxy)benzaldehyde

[0236] Into a 25-mL round-bottom flask, was placed (S)-2-((4-(2-(((tertbutyldimethylsiiyl)oxy)methyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybenzaldehyde (380 mg, 0.78 mmol, 1.0 equiv). To the above, HCl (g) (2 M) in EA (5 mL) was added at 0 °C. The reaction solution was stirred for 1 h at room température. The reaction was then quenched 20 by the addition of water (10 mL), and the pH value of the solution was adjusted to 8 with

NaHCO? solid and extracted with 3x10 mL of ethyl acetate. The combined organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC with the following conditions (2#SHIMADZU (HPLC-01)): Column, Kinetex EVO C18 Column, 21.2*150, 5 um, mobile 25 phase, Water (0.1% Formic Acid) and CH^CN (10% Phase B up to 50% within 15 min), detector, UV 254 nm. (S)-2-hydroxy-6-((4-(2-(hydroxymethyl)nicotinoyl)morpholin-3yl)methoxy)benzaldehyde was obtained. LCMS: (ES, [M+H]*: 373.1; [M+Na]⁺: 395.1.

Alternative Synthesis: Scheme 9B.

[0237] Alternative!y, Compound 12 can be synthesized as shown in Scheme 9B using similar 30 procedures described in Scheme 9A.

Scheme 9B

[0238] Compound 9c can be converted into 9d using methods known in the art (for example, sodium tetrahydroborate; acetic acid in tetrahydrofuran at 15 °C; for 4h). Then, using a silyl protecting group such as TBDPS (tert-butyldiphenylsily 1), intermediate 8e2 can be converted into compound 12 using similar conditions as described in Scheme 9Æ

Example 10. (S)-2-hydroxy-6-((4-(2-(hydroxymethyl)nicotinoyl)thiomorpholin-3yl)methoxy)benzaldehyde and (R)-2-hydroxy-6-((4-(2(hydroxymethyl)nkotinoyl)thiomorpholin-3-yl)methoxy)benzaldehyde

Scheme 10A

10b

OTBS OTBS

Enantiomers 1 and 2 of 10b

Compound 13 Enantiomers 1 and 2

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

[0239] To a solution of 2-[[(tert-butyldimethy]silyl)oxy]methyl]pyridine-3-carboxyIic acid (1.20 g, 4.50 mmol, 1.00 equiv) and thiomorpholin-3-ylmethanol hydrochloride (912 mg, 5.40 10 mmol, 1.20 equiv) in DMF under nitrogen at 0 °C was added DIEA (909 mg, 9.00 mmol, 2.00 equiv). This was followed by the addition of HATU (2.05 g, 5.40 mmol, 1.20 equiv) in several batches at 0 °C. The mixture was allowed to slowly warm to room température and stirred for 16 h. The reaction was diluted with water (100 mL) and extracted with 3 x 100 mL of ethyl acetate.

P The combined organic layer was washed with 3 x 50 mL of brine, dried over anhydrous sodium sulfate, and concentrated. The residue was applied onto a silica gel column and eluted with ethyl acetate/petroleum ether (from 0% to 100% ethyl acetate). Removal of the solvents produced [4(2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridine-3-carbonyl)thiomorpholin-3-yl]methanol.

LCMS (ES) [M+l] ⁺ m/z: 383.

Step 2. Synthesis of 2-((4-(2-(((tertbutyldimethylsilyl)oxy)methyl)nicotίnoyl)thΐomorpholiπ-3-yl)methoxy)-6hydroxybenzaldehyde

[0240] To a mixture of [4-(2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridine-310 carbonyl)thiomorpholin-3-yl]methanoI (1.20 g, 3.14 mmol, 1.00 equiv), 2,6dîhydroxybenzaldehyde (519 mg, 3.76 mmol, 1.20 equiv), and PPh₃ (0.99 g, 3.76 mmol, 1.20 equiv) in THF (50.0 mL) under nitrogen at 0 °C was added a solution of DBAD (0.87 g, 3.76 mmol, 1.20 equiv) in THF (1 mL) dropwise over 15 min. The resulting mixture was stirred for 16 h at room température. The reaction mixture was concentrated to remove solvents, and the 15 residue was applied onto a silica gel column, eluted with ethyl acetate/petroleum ether (1:0).

After removing solvents, this produced 2-[[4-(2-[[(tert-butyidimethylsilyl)oxy]methyl]pyridine3-carbonyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde. LCMS (ES) [M+l] ⁺ m/z: 503.

Step 3. Chiral-HPLC séparation of Compound 10b.

[0241] Racemic 2-[[4-(2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridine-3carbonyl)lhiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde was purified by Chiral-PrepHPLC with the following conditions: Agela HP-Flash (model: HP-1000); Mobile phase: A:nHexane/DCM=5/1; B: Ethanol; Flow rate: 30mL/min; Column: CHIRALPAK IG-3, 4.6*50 mm, 3pm; and Gradîent:20%B in 15min; 220nm.

[0242] This resulted in each of Enantiomers 1 and 2 of Compound 10b (Rt = 10 min and 12 min, respectively). LCMS (ES) [M+l] ⁺ m/z: 503.

Step 4a. Removal of TBS group to give Compound 13, Enantiomer L

[0243] To a solution of Enantiomer 1 of Compound 10b (119 mg, 0.24 mmol, 1.00 equiv) in THF (10.0 mL) was added triethylamine trihydrofluoride (“TEA.3HF”) (458 mg, 2.84 mmol, 30 12.0 equiv). The reaction was stirred for 16 h at room température. Solvents were removed, and the residue was applied onto a C18 silica gel column with Phase A: Water/0.05% TFA, Mobile

Phase B: Acetonitrile; Flow rate: 1.5 mL/min; Gradient: 5%B to 100%B in 1.2 min, hold 0.6 min. This resulted in Compound 13, Enantiomer 1. RT = 3.614 mins; LCMS (ES, m/z): [MH]⁺ 389.1.

Step 4b. Removal of TBS group to give Compound 13, Enantiomer 2.

[0244] To a solution of Enantiomer 2 of Compound 10b (120 mg, 0.24 mmol, 1.00 equiv) in

THF (10.0 mL) was added TEA.3HF (461 mg, 2.87 mmol, 12.0 equiv). The mixture was stirred for 16 h at room température. Solvents were removed, and the residue was applied onto a C18 silica gel column with Phase A: Water/0.05% TFA, Mobile Phase B: Acetonitrile; Flow rate: 1.5 mL/mîn; Gradient: 5%B to 100%B in 1.2 min, hold 0.6 min. This resulted in Compound 13,

Enantiomer 2. RT = 4.387 mins; LCMS (ES, m/z): [MH]⁺ 389.1; [MNa]⁺ 411.1.

Alternative synthesis of (R)-2-hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl)thiomorphoIin3-y l)methoxy )ben zaldehyde

[0245] Alternatively, (R)-2-hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl)thiomorpholin-3yl)methoxy)benzaldehyde can be made directly from chiral (R)-thiomorpholin-3-ylmethanol as 15 depîcted in Scheme 10B.

I

Scheme 10B

Step 1

[0246] Into a 2-L three-necked round-bottom flask, was placed a solution of (3-bromopyridin5 2-yl)methanol (50 g, 0.267 mol, l.ü equiv) in DCM (1.0 L) and IH-imidazole (36.4 g, 0.534 mol, 2.0 equiv). After the mixture was cooled to 0°C, tert-butyl(chloro)dimethylsilane (48.1 g, 0.320 mol, 1.2 equiv) was added by three batches. The reaction solution was warmed to room température and stirred for 4 h. The reaction mixture was diluted with H₂O (1.0 L) and extracted with 2x500 mL of DCM. The combined organic phase was dried over anhydrous sodium sulfate and filtered. The fikrate was concentrated under reduced pressure, and the residue was purified by silica gel column with ethyl acetate/petroleum ether (1:10) to provide the title compound. LCMS (ES) [M+l]⁺ m/z 3O2.d

Step 2

[0247] Into a 2-L three-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed a solution of 3-bromo-2-(((tertbutyldimethylsilyl)oxy)methyl)pyridine (70.0 g, 0.233 mol, LO equiv) in THF (700 mL). Thïs was followed by the addition of /i-BuLi (2.5 M in hexane) (102.5 mL, 0.256 mol, 1.1 equiv) dropwîse with stirring at -78°C. After addition, the mixture was stirred for 0.5 h, and ethyl carbonochloridate (37.8 g, 0.350 mol, 1.5 equiv) was added at the sanie température and stirred for 1 h. The reaction was then quenched by the addition of 500 mL of aqueous NH4CI and extracted with 2x600 mL of ethyl acetate. The combined organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1:4) to provide the title compound. LCMS (ES) [M+l]⁺ m/z 296.

Step 3

[0248] Into a 1-L three-necked round-bottom flask, was placed ethyl 2-(((tert10 butyldimethylsilyl)oxy)methyi)nicotinate (38.6 g, 0.131 mol, 1.0 equiv), MeOH (400 mL), and H2O (200 mL). This was followed by the addition of LiOHThO (11.0 g, 0.262 mol, 2.0 equiv) at 0^üC. The mixture was stirred for 2 h at room température. The mixture was concentrated to remove the solvent, and the pH value of the residue was adjusted to 7 with citric acid. The solution was filtered, and the solid was dried under infrared iamp. 2-(((tert15 butyldimethylsilyl)oxy)methyl)nicotinic acid was obtained. LCMS (ES) [M+l]⁺ m/z: 268.

Step 4

[0249] Compound 6h was prepared as described in Scheme 6B. To a solution of 2-[[(tertbutyldimethylsilyl)oxy]methyl]pyridine-3-carboxylic acid (10.0 g, 37.3 mmol, 1.0 equiv), DIPEA (12.1 g, 93.5 mmol, 2.5 equiv) and HATU (17.06 g, 44.877 mmol, 1.20 equiv) in DMF (100 mL) was added (3R)-thiomorpholin-3-ylmethanol (4.98 g, 37.397 mmol, 1.00 equiv) at 0 ⁿC in portions. The resulting solution was stirred for 4 hr at 0-25 °C. The reaction was then quenched by the addition of 200 mL of water. The resulting solution was extracted with 3x200 mL of ethyl acetate, dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was appiied onto a silica gel column with ethyl acetate/petroleum ether (1:100 to 1:10).

This resulted in [(3R)-4-(2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridine-3carbonyl)thiomorpholin-3-yl]methanol. LCMS (ES) [M+l] ⁺ m/z: 383.2; Rétention time 1.138 min. Ή-NMR: (300 MHz, CDC13, ppm): δ 8.60 (dd, J =4.8,1.5 Hz, 1H), 7.67-7.53 (m, 1H), 7.29-7.25 (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).

Step 5

[0250] A solution of [(3R)-4-(2-[[(tert-butyldimethylsîlyl)oxy]methyl]pyridine-3carbonyl)thiomorpholin-3-yl]methanol (11.0 g, 28.7 mmol, 1.0 equiv), 2,6dihydroxybenzaldehyde (4.7 g, 34.5 mmol, 1.2 equîv) and PPh3 (9.8 g, 37.3 mmol, 1.3 equiv) in 5 DCM (1.1 L) was cooled to 0 °C under Ar atmosphère. A solution of DBAD (7.28 g, 230.2 mmol, 1.1 equiv) in DCM (100 mL) was added dropwise. The resulting solution was stirred for 16 hr at 0-25 °C. The réaction was concentrated under vacuum. The residue was applied onto a silîca gel column with ethyl acetate/petroleum ether (1:100 to l:5).This resulted in 2-[[(3R)-4(2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridine-3-carbonyl)thiomorpholin-3-yl]methoxy]-610 hydroxybenzaldehyde. LCMS (ES) [M+l] ⁺ m/z: 503.2; Rétention time 1.223 min. 'H-NMR: (300 MHz, DMSO-(A, ppm): δ 11.74 (br, 1H), 10.24 (br, 1H), 8.59 (dd, J = 4.9, 1.7 Hz, 1H), 7.88-7.41(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).

Step 6

[0251] Into a 500-mL 3-necked round-bottom flask, was placed 2-[[(3R)-4-(2-[[(tertbutyIdimethylsilyl)oxy]methyl]pyridine-3-carbonyl)thiomorpholin-3-yl]methoxy]-6hydroxybenzaldehyde (13.6 g, 27.0 mmol, 1.0 equiv) and THF (150 mL). After the reaction was cooled to 0°C, a solution of TEA 3HF (13.0 g, 80.9 mmol, 3.0 equiv) was added dropwise. The resulting solution was stirred for 5 h at 0-25 °C. The pH value of the solution was adjusted to 8 with NaHCOa (2 mol/L). The resulting solution was extracted with ethyl acetate (200 mLx3), and the organic layers combiued and concentrated. The crude product was purified by FlashPrep-HPLC with the following conditions (IntelFlash-1): Column, C18 silica gel; mobile phase, MeCN=10/90 increasîng to MeCN=90/10; Detector, 220. This resulted in 2-hydroxy-6-[[(3R)-4[2-(hydroxymethyl)pyridine-3-carbonyl]thiomorpholin-3-yl]methoxy]benzaldehyde. LCMS (ES, m/z)·. [M+FI] ⁺: 389.1; Rétention time 1.060 min.

[0252] Analytical SFC rétention time: 3.641 min. Conditions for SFC were as follows: Instrument Name: Shimadzu LC30AD SF; Column: OD-3, 100*3.0 mm, 3 um; Column ID: OD3SCK-TG002; Oven Température: 35 C; Total Flow: 2.5000 mL/min; Start Conc. of Pump B: 10.0%; BPR Pressure: 15.00 MPa.

[0253] ‘H-NMR (300 MHz, DMSO-i/6, ppm): δ H .77 (br, 1H), 10.30 (br, 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).

[0254] Based on the product of Scheme 10B, it was determined that Compound 13, Enantîomer 1 corresponds to (R)-2-hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl)thiomorpholin3-yl)methoxy)benzaldehyde.

Example 11. (S)-2-hydroxy-6-((1-(2-(2-methoxyethyl)benzoyl) pi per îdin-2 5 yl)methoxy)beuzaldehyde, Compound 14

[0255] Compound 14 was synthesized according to Scheme 11.

Scheme 11

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

[0256] Into a 500-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen was placed 2-(2-bromophenyI)ethanol (10.0 g, 49.7 mmol, 1.00 equiv) and DMF (100 mL) cooled to 0 °C by ice water, and then NaH (2.39 g, 99.5 mmol, 2.00 equiv) was added in several portions. The resulting solution was stirred for 40 min at 0 °C, and then Mel (10.59 g, 74.610 mmol, 1.50 equiv) was added dropwise with stirring at 0 °C over 15 mins.

The resulting solution was allowed to warm up to room température with stirring for an additional 5 hr. The reaction was then quenched by the addition of water/ice. The resulting solution was extracted with 3 x 100 mL of ethyl acetate and the organic layers were combined and dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by silica gel column chromatography with ethyl acetate/hexane (1:3) as eluents. This resulted in l-bromo-2-(2-methoxyelhyl)benzene. LCMS (ES) [M+l]⁺ m/z: 215.

” Step 2. Synthesis of ethyl 2-(2-methoxyethyl)benzoate

[0257] Into a 500-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen was placed l-bromo-2-(2-methoxyethyl)benzene (10.0 g, 46.5 mmol, 1.00 equiv) and THF (100 mL). The mixture was cooled to -78 °C, and n-butyllithium (39 mL, 5 97.7 mmol, 2.10 equiv) was added dropwise into the solution. The resulting solution was stirred for 40 min at -78 °C, then ethyl chloroforma te (7.57 g, 69.757 mmol, 1.50 equiv) was added dropwise. The resulting solution was brought to room température with stirring for an additional 5 min at -78 °C and then stirred for an additional 16 hr al room température. The reaction was then quenched by the addition of water/ice. The resulting solution was extracted with 3 x 100 10 mL of ethyl acetate, and the organic layers were combined and dried over anhydrous sodium sulfate. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1:5) as eluent. This resulted in ethyl 2-(2-methoxyethyl)benzoate. LCMS (ES) [M+l]⁺ m/z: 209.

Step 3. Synthesis of 2-(2-methoxyethyl)benzoie acid

[0258] Into a 100-mL round-bottom flask was placed ethyl 2-(2-methoxyethyl)benzoate (1.20 g, 5.76 mmol, 1.00 equiv), LiOH (0.55 g, 23.0 mmol, 4.00 equiv) and THF (15.0 mL), and HjO (3.00 mL). The resulting solution was stirred for 4 hr at 50 °C in an oil bath. The reaction mixture was cooled with a water/ice bath. The pH of the solution was adjusted to 5 with HCl (2M). The resulting solution was extracted with 3 x 50 mL of ethyl acetate, and the organic layers were combined and dried over anhydrous sodium sulfate and concentrated under vacuum.

This resulted in 2-(2-methoxyethyl)benzoic acid. LCMS (ES) [M+l] ⁺ m/z: 181.

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

[0259] Into a 250-mL round-bottom flask was placed 2-(2-methoxyethyl)benzoic acid (550 mg, 3.05 mmol, 1.00 equiv), (2S)-piperidin-2-ylmethanol (421 mg, 3.66 mmol, 1.20 equiv), 25 HATU (2.32 g, 6.10 mmol, 2.00 equiv), DIEA (788 mg, 6.10 mmol, 2.00 equiv) and DCM (40.00 mL). The resulting solution was stirred for 4 hr at room température. The resulting solution was extracted with 3 x 30 mL of dichloromethane, and the organic layers were combined and dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by silica gel column with ethyl acetate/petroleum ether (2:5) as eluents. This resulted in [(2S)-l-[2-(2-methoxyethyl)benzoyl]piperidin-2-yl]methanol. LCMS (ES) [M+l] ⁺ m/z: 278.

100

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

[0260] Into a 100-mL round-bottom flask purged and maintained with an inert atmosphère of nitrogenwas placed [(2S)-l-[2-(2-methoxyethyl)benzoyl]piperidin-2-yl]methanol (470 mg, 1.70 mmol, 1.00 equiv), 2,6-dîhydroxybenzaldehyde (468 mg, 3.39 mmol, 2.00 equiv), PPh₃ (888 mg, 3.39 mmol, 2.00 equiv) and THF (30.0 mL). The resulting solution was stirred for 15 min at 0 °C, and then DIAD (685 mg, 3.39 mmol, 2.00 equiv) was added dropwise. The resulting solution was stirred for 15 min at 0 °C. The resulting solution was warmed up to room température with stirring for an additional 16 hr. The resulting solution was extracted with 3 x

30 mL of ethyl acetate, and the organic layers were combined and dried over anhydrous sodium sulfate. The crude product was purified by Prep-HPLC [Column: Atlantis HILIC OBD Column, 19* 150mm *5um; mobile phase: Waler(0.1%FA) and ACN (10% Phase B up to 50% in 10 min, up to 90% in 10 min)]. This resulted in (S)-2-hy droxy-6-((1-(2-(2methoxyethyI)benzoyl)piperidin-2-yl)methoxy)benzaldehyde. LCMS (ES) [M+l] + m/z: 398.

Ή NMR (300 MHz, DMSO-d6) 11.64 (br, 1H), 10.29 (br, 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), 1.921.34 (m, 6H).

101

Example 12. (S)-3-(2-(2-((2-formyl-3-hydroxyphenoxy)methyl)piperidme-lcarbonyl)phenyl)propanenitrile, Compound 15

[0261] Compound 15 was synthesized according to Scheme 12.

Scheme 12

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

[0262] Into a 100-mL round-bottom flask, was placed methyl 2-bromobenzoate (5.00 g, 23.251 mmol, 1.00 equiv), acrylonitrile (12.34 g, 232.508 mmol, 10.00 equiv), DIEA (6.01 g, 46.502 mmol, 2.00 equiv), and bis(tributylphosphine) palladium (1.19 g, 2.325 mmol, 0.10 equiv). The resulting solution was stirred for 16 hr at 80 °C. The resulting mixture was concentrated. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:5) as eluents. The collected fractions were combined and concentrated. This resulted in methyl 2-[(lE)-2-cyanoeth-l-en-l-yl]benzoate. GCMS M⁺: 187.

102

Step 2. Synthesis of methyl 2-(2-cyanoethyI)benzoate

[0263] Into a 100-mL round-bottom flask, was placed methyl 2-[(lE)-2-cyanoeth-l-en-ly]]benzoate (2.40 g, 12.8 mmol, 1.00 equîv), methanol (50 mL), and Pd/C (0.24 g). The flask was evacuated and flushed three times with nitrogen, followed by flushing with hydrogen. The 5 mixture was stirred 16 h at room température under an atmosphère of hydrogen. The solids were fîltered out. The resulting mixture was concentrated. This resulted in methyl 2-(2cyanoethyl)benzoate. LCMS (ES) [M+l]+ m/z 190.1.

Step 3. Synthesis of 2-(2-cyanoethyI)benzoic acid

[0264] Into a 100-mL round-bottom flask was placed methyl 2-(2-cyanoethyl)benzoate (2.20 10 g, 11.627 mmol, 1.00 equiv), methanol (20 mL), water (20 mL) and sodium hydroxide (0.93 g, 23.252 mmol, 2.00 equiv). The resulting solution was stirred for 4 hr at 25 °C. The reaction was then quenched by the addition of 100 mL of water. The pH value of the solution was adjusted to 5 with HCl (1 mol/L). The solids were coliected by filtration. This resulted in 2-(2cyanoethyl)bcnzoic acid. LCMS (ES) [M-l] m/z 174.1.

Step 4. Synthesis of 3-[2-[(2S)-2-[[(tert-butyldimethylsilyl)oxy]methyI]piperidine-lcarbonyl]phenyl]propanenitrile

[0265] Into a 100-mL round-bottom flask was placed 2-(2-cyanoethyI)benzoic acid (1.80 g, 10.275 mmol, 1.00 equiv), DCM (30.00 mL), (2S)-2-[[(tertbutyldimethylsilyl)oxy]methyl]piperidine (2.36 g, 10.275 mmol, 1.00 equiv), HATU (5.86 g, 20 15.412 mmol, 1.50 equiv) and DIEA (3.98 g, 30.824 mmol, 3.00 equiv). The resulting solution was stirred for 16 hr at 25 ^ÛC. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1:2) as eluents. The coliected fractions were combined and concentrated. This resulted in 3-[2-[(2S)-2-[[(tertbutyldimethylsilyl)oxy]methyl]piperidîne-l-carbonyl]phenyl]propanenitrile. LCMS (ES)

[M+l]* m/z 387.2.

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

[0266] Into a 100-mL round-bottom flask, was placed 3-[2-[(2S)-2-[[(tertbutyldimethylsilyl)oxy]methyl]piperidîne-l-carbonyl]phenyl]propanenitrile (2.00 g, 5.173 mmol, 1.00 equiv), tetrahydrofuran (20 mL) and TBAF (0.27 g, 1.035 mmol, 0.20 equiv). The resulting solution was stirred for 2 hr at 25 ^UC. The resulting mixture was concentrated. The

103 residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1:2) as eluents. The collected fractions were combined and concentrated. This resulted in 3-[2[(2S)-2-(hydroxymethyl)piperidinc-l-carbonyl]phenyl]propanenitrîle. LCMS (ES) [M+l]⁺ m/z 273.2.

Step 6. Synthesis of (S)-3-(2-(2-((2-formyl-3-hydroxyphenoxy)methyl)piperidîne-lcarbonyl)phenyl)propanenitrile

[0267] Into a 50-mL round-bottom flask, was placed 3-[2-[(2S)-2-(hydroxymethyl)piperidinel-carbonyl]phenyl]propanenitrile (100.00 mg, 0.367 mmol, 1.00 equiv), tetrahydrofuran (8.00 mL), 2,6-dihydroxybenzaldehyde (50.72 mg, 0.367 mmol, 1.00 equiv), PPh3 (115.57 mg, 0.441 mmol, 1.20 equiv), and DIAD (89.10 mg, 0.441 mmol, 1.20 equiv). The resulting solution was stirred for 16 hr at 25 °C. The resulting mixture was concentrated under vacuum. The crude reaction mixture was filtered and subjected to reverse phase préparative HPLC (Prep-C18, 2045M, 120 g, Tianjin Bonna-Agela Technologies; gradient elution of 30% MeCN in water to 40% MeCN in water over a 10 min perîod, where both solvents contain 0.1% FA) to provide (S)-3 -(2-(2-((2-formy 1 -3 -hy droxyphenoxy )methyl)piperîdine-1 -carbony l)pheny l)propanenitrile.

LCMS (ES) [M+l]⁺ m/z 393.2. 1H NMR (300 MHz, DMSO-d6) δ 1 1.72 (br, 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).

104

Example 13. (S)-2-hydiOxy-6-((l-(3-(2-hydroxyethyl)picoIinoyl)piperidin-2’ yl)methoxy)benzaldehyde, Compound 16

[0268] Compound 16 was synthesized according to Scheme 13.

Scheme 13

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

[0269] Into a 100-mL round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed 2-[(Z)-2-ethoxyethenyl]-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (4.13 g, 20.S5 mmol, 1.50 equiv), methyl 3-bromopyridine-2-carboxylate (3.00 g, 13.89 mmol, 1.00 equiv), dioxane (30.00 mL), H2O (6.00 mL), NaaCOr (4.42 g, 41.66 mmol, 3.00 equiv), and Pd(PPhr)4 (1.60 g, 1.39 mmol, 0.10 equiv). The resulting solution was stirred overnight at 80 ^ÛC. The reaction mixture was cooled to room température. The solids were filtered out, and the filtrate was concentrated. The residue was purified by silica gel column chromatography with THF/PE (30%) as eluent. This resulted in methyl 3-[(E)-2-ethoxyethenyl]pyridine-215 carboxylate. LCMS (ES) [M+l]⁺ m/z 208.

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

[0270] Into a purged 100-mL round-bottom flask was placed methyl 3-[(E)-2ethoxyethenyl]pyridine-2-carboxylate (2.60 g, 12.55 mmol, 1.00 equiv), Pd/C (500.00 mg, 4.69 mmol, 0.37 equiv), and MeOH (30.00 mL). The flask was evacuated and flushed three times with nitrogen, followed by flushing with hydrogen. The resulting solution was stirred overnight

105 at room température. The solids were filtered out, and the filtrate was concentrated. This resulted in methyl 3-(2-ethoxyethyl)pyridine-2-carboxylate. LCMS (ES) [M+l]⁺ m/z 210.

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

[0271] Into a 100-mL round-bottom flask, was placed methyl 3-(2-ethoxyethyl)pyrîdine-25 carboxylate (2.50 g, 11.95 mmol, 1.00 equiv), THF (25.00 mL), H₂O (5.00 mL), and LiOH H₂O (1.00 g, 23.83 mmol, 1.99 equiv). The resulting solution was stirred for 2 h at room température. The resulting mixture was concentrated and was adjusted to pH 3-4 with HCl (1 mol/L). The resulting solution was extracted with 3x20 mL of ethyl acetate, and the organic layers were combined and dried over anhydrous sodium sulfate and concentrated. This resulted in 3-(210 ethoxyetbyi)pyridine-2-carboxylic acid. LCMS (ES) [M+l]⁺ m/z 196.

Step 4. Synthesis of 2-[(2S)-2-[[(tert-butyldimethylsilyl)oxy]methyl]piperidine-l-carbonylJ3-(2-ethoxyethyl)pyridine

[0272] Into a 100-mL 3-necked round-bottom flask was placed (2S)-2-[[(tertbutyldimethylsilyl)oxy]methyl]piperidme (2.12 g, 9.22 mmol, 1.00 equiv), 3-(215 ethoxyethyl)pyridine-2-carboxylic acid (1.80 g, 9.22 mmol, 1.00 equiv), DCM (20.00 mL), EhN (1.87 g, 18.44 mmol, 2.00 equiv), EDCI (2.12 g, 11.06 mmol, 1.20 equiv), and HOBt (1.50 g, 11.06 mmol, 1.20 equiv). The resulting solution was stirred overnight at room température. The reaction was then quenched by the addition of 10 mL of water. The resulting solution was extracted with 3x20 mL of dichloromethane, and the organic layers were combined and dried over anhydrous sodium sulfate and concentrated. This resulted in 2-[(2S)-2-[[(terlbutyIdimethylsilyl)oxy]methyl]piperidine-l-carbonyl]-3-(2-ethoxyethyl)pyridine. LCMS (ES) [M+l]⁺ m/z 407.

Step 5. Synthesis of [(2S)4-[3-(2-ethoxyethyl)pyridine-2-carbonyI]piperidin-2-yl]methanol ।

[0273] Into a 100-mL round-bottom flask, was placed 2-[(2S)-2-[[(tert25 butyldimethylsilyl)oxy]methyl]piperidine-l-carbonyi]-3-(2-ethoxyethyl)pyridine (3.00 g, 7.38 mmol, 1.00 equiv), THF (20.00 mL), and TBAF/THF (14.75 mL, 14.75 mmol, 2.00 equiv). The resulting solution was stirred overnight at room température. The resulting mixture was concentrated. The residue was purified by silica gel column with THF/PE (45%) as eluent. This resulted in [(2S)-l-[3-(2-ethoxyethyl)pyridine-2-carbonyl]piperidin-2-yl]methanol. LCMS (ES)

[M+l]⁺ m/z 293.

106

Step 6. Synthesis of 2-[[(2S)-l-[3-(2-ethoxyethyl)pyridine-2-carbonyI]piperidin-2yl]methoxy]-6-hydroxybenzaldehyde

[0274J Into a 100-mL 3-necked round-bottom flask, was placed [(2S)-l-[3-(2ethoxyethyl)pyridine-2-carbonyl]pîperidin-2-yl]methanol (1.70 g, 5.81 mmol, 1.00 equiv), 2,65 dihydroxybenzaldehyde (0.96 g, 6.95 mmol, 1.20 equiv), DCM (40.00 mL), and PPM (1.83 g, 6.98 mmol, 1.20 equiv). This was followed by the addition of DIAD (1.41 g, 6.98 mmol, 1.20 equiv) dropwîse with stirring at 0 °C. The resulting solution was stirred for 4 h at room température. The resulting mixture was concentrated. The residue was purified by silica gel column with THF/PE (30%) as eluents. This resulted in 2-[[(2S)-l-[3-(2-ethoxyethyI)pyridine-210 carbonyl]piperidin-2-yl]methoxy]-6-hydroxybenzaldehyde. LCMS (ES) [M+l]⁺ m/z 413.

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

[0275] Into a 100-mL 3-necked round-bottom flask, was placed 2-[[(2S)-l-[3-(2ethoxyethyl)pyrîdine-2-carbonyl]piperidin-2-yI]methoxy]-6-hydroxybenzaldehyde (500.00 mg, 15 1.21 mmol, 1.00 equiv) and DCM (20.00 mL). This was followed by the addition of BBr₃/DCM (12.12 mL, 12.12 mmol, 10.00 equiv) dropwîse with stirring at -78 °C. The resulting solution was stirred for 1 h at 0 °C. The reaction was then quenched by the addition of 20 mL of water/ice. The resulting solution was extracted with 3x20 mL of dichloromethane, and the organic layers were combined and dried over anhydrous sodium sulfate and concentrated. The 20 crude product was purified by Prep-HPLC [Column, XBridge Prep C18 OBD Column, 19cm,

150mm, 5um; mobile phase, Water (0.1% HCOOH) and AcCN (30% Phase B up to 60% in 11 min); Detector, 254. This resulted in (S)-2-hydroxy-6-((l-(3-(2hydroxyethyl)picoIinoyl)piperîdin-2-yl)methoxy)benzaldehyde. LCMS: (ES, m/z): [M+l] ⁺ 385.0. Ή-NMR (300 MHz, DMSO-d6) δ l l .81 (s, 1 H), 10.29 (d, J = 6.6 Hz, 1 H), 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), 3.67 - 3.56 (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).

107

Example 14. (S)-2“hydrüxj'6-((l-(2-(2-(pyrrolidin-l-yl)ethyI)benzoyl)piperidin-2yl)methoxy)benzaldehyde, Compound 17

[0276] Compound 17 was synthesized according to Scheme 14.

Scheme 14

Step 1. Synthesis of 2-bromO'3-[[(2S)-l-[2-[(E)-2-ethoxyethenyI]benzoyl]piperidin-2yl]methoxy]phenol

[0277] Into a 100-mL round-bottom flask purged and maintained with an inert atmosphère of nitrogen was placed a solution of [(2S)-l-[2-[(E)-2-elhoxyethenyl]benzoyl]piperidin-210 yl]methanol (5.00 g, 0.017 mmol, 1.00 equiv) in THF (50 mL), 2-bromobenzene-l,3-diol (3.27 g, 0.017 mmol, 1 equiv), DIAD (4.19 g, 0.021 mmol, 1.2 equiv), and PPh₃ (5.44 g, 0.021 mmol, 1.2 equiv). The resulting solution was stirred for 1 hr at 0 °C in an ice/salt bath, then was removed from the bath and allowed to stir overnight at room température. The reaction was then quenched by the addition of water/îce. 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 with ethyl acetate/petroleum ether (1:4) as eluent. This resulted in 2-bromo-3-[[(2S)-l -[2-[(E)-2-ethoxyethenyI]benzoyl]piperidîn-2-yl]meihoxy]phenol. LCMS (ES) [M+l]+ m/z 460.2.

108

Step 2. Synthesis of2-[2-[(2S)-2-(2-bromo-3-hydroxyphenoxymethyl)piperidine-lcarbonyl] phenyl] acétaldéhyde

[0278] Into a 100-mL round-bottom flask, was placed 2-bromo-3-[[(2S)-l-[2-[(E)-2ethoxyethenyl]benzoyl]piperidin-2-yI]methoxy]phenol (3,00 g, 6.517 mmol, 1.00 equiv) and IM 5 HCl in EtOAc (20 mL). The resulting solution was stirred overnight at room température. The resulting mixture was concentrated under vacuum. This resulted in 2-[2-[(2S)-2-(2-bromo-3hydroxyphenoxymethyl)piperidine-l-carbonyl]phenyl]acetaldehyde. LCMS (ES) [M+l]⁺ m/z 432.2.

Step 3. Synthesis of 2-bromo*3-[[(2S)-l-[2-[2-(pyrrolidin-l-yl)ethyl]benzoyI]piperidin-2 10 yl]methoxy] phénol

[0279] Into a 100-mL round-bottom flask, was placed a solution of 2-[2-[(2S)-2-(2-bromo-3hydroxyphenoxymethyi)piperidine-l-carbonyl]phenyl]acetaldehyde (2.20 g, 0.005 mmol, 1.00 equiv) in MeOH (10 mL), pyrrolidine (0.54 g, 0.008 mmol, 1.5 equiv), MeOH (1 equiv), and NaBH₄ (0.48 g, 0.013 mmol, 2.5 equiv). The resulting solution was stirred for 1 hr at 0 °C in an ice/salt bath. The resulting solution was stirred overnight at room température. The reaction was then quenched by the addition of 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 with dichloromethane/methanol (1:10) as eluents. This resulted in 2-bromo-3-[[(2S)-l-[2-[2-(pyrrolidin-l-yI)ethyl]benzoyl]piperidin-220 yl]methoxy]phenol. LCMS (ES) [M+l]⁺ m/z 487.2.

Step 4. Synthesis of 2-ethenyl-3-[[(2S)-l-[2-[2-(pyrroHdin-l-yl)ethyl]benzoyl]piperidin-2yl]methoxy] phénol

[0280] Into a 25-mL round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed 2-bromo-3-[[(2S)-l-[2-[2-(pyrroIidîn-l-yl)ethyI]benzoyl]piperidin-225 yl]methoxy]phenol (600.00 mg, 1.231 mmol, 1.00 equiv) in dioxane (10 mL), 2-ethenyl-4,4,5,5tetramethyl-l,3,2-dioxaborolane (189.59 mg, 1.231 mmol, 1 equiv), K2CO3 (340.24 mg, 2.462 mmol, 2 equiv), and Pd(dppf)C12 (45.03 mg, 0.062 mmol, 0.05 equiv). The resulting solution was stirred overnight at 80 °C in an oil bath. The reaction was then quenched by the addition of water. The resulting solution was extracted with lOOmL of dichloromethane, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (2:1) as eluent. This resulted in 2-ethcnyl-3

109

[[(2S)-l-[2-[2-(pyrrolidin-l-yl)ethyl]benzoyl]piperidin-2-yl]methoxy]phenol. LCMS (ES) [M+l]⁺m/z 435.1.

Step 5. Synthesis of (S)-2-hydroxy-6-((l-(2-(2-(pyrrolidin-l-yl)ethyl)benzoyl)piperidin-2y I) methoxy )benzaldehy de

[0281] Into a 10-mL round-bottom flask, was placed a solution of 2-ethenyl-3-[[(2S)-l-[2-[2(pyrroiidin-l-yl)ethyl]benzoyl]pipendin-2-yl]methoxy]phenol (120.00 mg, 0.276 mmol, 1.00 equiv) in acetone (5 mL), a solution of NaIO₄ (118.12 mg, 0.552 mmol, 2 equiv) in H₂O (2 mL), and K2OSO4 2H2O (10.17 mg, 0.028 mmol, 0.1 equiv). The resulting solution was stirred overnight at room température. The solids were filtered out, and the filtrate was concentrated.

The residue was purified by silica gel column chromatography with ACN: H₂O (1:4) as eluent. This resulted in (S)-2-hydroxy-6-((l-(2-(2-(pynOlidin-l-yl)ethyl)benzoyI)piperidin-2yl)methoxy)benza!dehyde. LCMS (ES) [M+l]⁺ m/z 437.3. 1H NMR (300 MHz, Chloroform-d) δ 12.02 (br, 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, 10H), 2.16-1.16 (m, 10H).

Example 15. (S)-2-hydroxy-6-((l-(3-(2-hydroxyethyl)pyrazine-2-carbonyl)pyrrolidin-2yl)methoxy)benzaldehyde, Compound 19

[0282] Compound 19 was synthesized according to Scheme 15.

Scheme 15

110

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

[0283] Into a 250-mL round-bottom flask purged and maintaîned with an inert atmosphère of nitrogen, was placed methyl 3-bromopyrazine-2-carboxylate (5.00 g, 23.04 mmol, 1.00 equiv), 2-ethenyl-4,4,5,5-tetramethyl-l,3,2-dioxaboroIane (4.26 g, 27.66 mmol, 1.20 equiv), dioxane (60.00 mL), H₂O (10.00 mL), K₂CO₃ (6.37 g, 46.08 mmol, 2.00 equiv), and Pd(dppf)Cl₂ (1.69 g,

2.30 mmol, 0.10 equiv). The resulting solution was stirred for 5 h at 80 °C. The reaction mixture was cooled to room température. The solids were filtered ont. The filtrate was concentrated, and the resulting residue was purîfied by sîlica gel column chromatography with THF/PE (15%) as eluent. This resulted in methyl 3-ethenylpyrazine-2-carboxylate. LCMS (ES) [M+l]⁺ m/z: 165.

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

[0284] Into a 250-mL round-bottom flask, was placed methyl 3-ethenyIpyrazine-2-carboxylate (3.50 g, 21.32 mmol, 1.00 equiv), MeOH (40.00 mL), and NaOMe (3.46 g, 64.05 mmol, 3.00 equiv). 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 with HCl (1 mol/L). The resulting mixture was concentrated. The residue was purîfied by silica gel column chromatography with MeOH/DCM (10%) as eluent. This resulted in 3-(2methoxyethyl)pyrazine-2-carboxylic acid. LCMS (ES) [M+l]⁺ m/z: 183.

Step 3. Synthesis of [(2S)-l-[3-(2-methoxyethyi)pyrazine-2-carbonyl]pyrrolidin-2yl|methanol

[0285] Into a 250-mL 3-necked round-bottom flask was placed 3-(2-methoxyethyl)pyrazine-2carboxylic acid (1.50 g, 8.23 mmol, 1.00 equiv), prolinol (0.83 g, 8.21 mmol, 1.00 equiv), DIEA (3.19 g, 24.70 mmol, 3.00 equiv), and DMF (30.00 mL). This was followed by the addition of HATU (3.76 g, 9.88 mmol, 1.20 equiv) in portions at 0 °C. The resulting solution was stirred overnight at room température. The crude product was purîfied by Prep-HPLC with the following conditions: Column, XBridge Prep C18 OBD Column, 19cm, 150mm, 5um; mobile phase, Water (0.1% HCOOH) and MeCN (5% Phase B up to 20% in 10min); Detector, 254. This resulted in [(2S)-l-[3-(2-methoxyethyl)pyrazine-2-carbonyI]pyrrolidin-2-yl]methanol. LCMS (ES) [M+lf m/z: 266.

111

Step 4. Synthesis of2-hydroxy-6-[[(2S)-l-[3-(2-methoxyethyl)pyrazine-2carbonyl]pyrrolidin-2-yl]methoxy]benzaldehyde

[0286] Into a 100-mL 3-necked round-bottom flask purged and maîntained with an inert atmosphère of mtrogen, was placed [(2S)-l-[3-(2-methoxyethyl)pyrazine-2-carbonyl]pyrrolidin5 2-yl]methanol (1.00 g, 3.77 mmol, 1.00 equiv), 2,6-dihydroxybenzaldehyde (0.62 g, 4.49 mmol, 1.19 equiv), PPh₃ (1.19 g, 4.52 mmol, 1.20 equiv), and DCM (30.00 mL). This was followed by the addition of DIAD (0.91 g, 4.52 mmol, 1.20 equiv) dropwise with stirring at 0 °C. The resulting solution was stirred overnight at room température. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with EA/DCM (10%) as eiuent. This resulted in 2-hydroxy-6-[[(2S)-l-[3-(2-methoxyethyl)pyrazine-2carbonyI]pyrroiidin-2-yl]methoxy]benzaldehyde. LCMS (ES, m/z): [M+H] ⁺: 386.2

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

[0287] Into a 100-mL 3-necked round-bottom flask, was placed 2-hydroxy-6-[[(2S)-1-(3-(215 methoxyethyl)pyrazine-2-carbonyl]pyrrolidin-2-yl]methoxy]benzaldehyde (360.00 mg, 0.93 mmol, 1.00 equiv) and DCM (10.00 mL). This was followed by the addition of BBr₃/DCM (9.34 mL, 9.34 mmol, 10.00 equiv) dropwise with stirring at -78 °C. The resulting solution was stirred for 2 h ai 0 °C. The reaction was then quenched by the addition of 20 mL of water/îce. The resulting solution was extracted with 3x20 mL of dichloromethane, and the organic layers were combined and dried over anhydrous sodium sulfate and concentrated. The crude product was purified by Prep-HPLC with the following conditions: Column, XBridge Prep C18 OBD Column, 19cm, 150mm, 5um; mobile phase, Water (0.1% HCOOH) and ACN (20% Phase B up to 50% in 11 min); Detector, 254. This resulted in (S)-2-hydroxy-6-((l-(3-(2hydroxyethyl)pyrazine-2-carbonyl)pyrrolîdîn-2-yl)methoxy)benzaldehyde. LCMS: (ES, m/z):

[M+H]⁺: 372. ’H-NMR: (300 MHz, DMSO-d6) δ 11.77 (s, 1 H), 10.34 (s, 1 H), 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), 3.93 - 3.63 (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).

112

Example 16. 2-hydroxy-6-{[(3S)-4-[2-(2-methoxyethyl)pyridine-3-carbonyl]morpholin-3 yl]methoxy}benzaldehyde, Compound 20

I

[0288] Compound 20 was synthesized according to Scheme 16.

Scheme 16

Step 1

[0289] Into a 250-mL round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed methyl 2-chloropyridine-3-carboxylate (10.0 g, 58.28 mmol, 1.0 equiv), dioxane (100 mL), tributyl(ethenyl)stannane (37.0 g, 116.56 mmol, 2.0 equiv), and Pd(dppf)C12 (4.26 g, 5.83 mmol, 0.1 equiv). The mixture was stirred for 12 h at 80 °C in oil bath. After cooled to room température, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1:3) as eluents. Methyl 2-ethenylpyridine-3-carboxylate was obtained. LCMS (ES) [M+l]+ m/z: 164.

Step 2

[0290] Into a 250-mL round-bottom flask, was placed methyl 2-ethenylpyridine-3-carboxylate (7.80 g, 47.90 mmol, 1.0 equiv) andMeOH (50 mL), HCl (c) (8.0 mL). The reaction solution was stirred for 12 h at 90 °C in oil bath. The reaction mixture was cooled to room température. Methyl 2-(2-methoxyethyl)pyridine-3-carboxylate hydrochloride was obtained and used in the next step directly without further purification. LCMS (ES) [M-HC1+1]+ m/z: 196.

113

Step 3

[0291] Into a 250-mL round-bottom flask, was placed rr ethyl 2-(2-methoxyethyl)pyridine-3carboxylate hydrochloride (7.0 g, 3Ü.3Ü mmol, 1.0 equiv), MeOH /H2O (1:2) (150 mL), and NaOH (2.40 g, 60.60 mmol, 2.0 equiv). The mixture was stirred for 2 h at 50 °C in oil bath.

After being cooled to room température, the solution was concentrated under reduced pressure. The pH value of the residue was adjusted to 6 with (6 M) HCl and purified by C18-120 g column with conditions: CH3CN/H2O from 5% increased to 60% within 12 min. 2-(2methoxyethyl)pyridine-3-carboxylic acid hydrochloride was obtained. LCMS (ES) [M-HCI+1]+ m/z: 182.

Step 4

[0292] Into a 50-mL 3-necked round-bottom flask purged and mamtained with an inert atmosphère of nitrogen, was placed 2-(2-methoxyethyl)pyridine-3-carboxylic acid hydrochloride (1.0 g, 4.60 mmol, 1.0 equiv), DMF(20 mL), DIEA (2.38 g, 18.40 mmol, 4.0 equiv), and (3R)morpholîn-3-yImethanoî hydrochloride (0.85 g, 5.51 mmol, 1.2 equiv). This was followed by the 15 addition of HATU (2.10 g, 5.51 mmol, 1.2 equiv) in several batches at 0°C. The reaction solution was stirred for 12 h at room température. The reaction solution was diluted with 30 mL of H2O and extracted with 3x100 mL of ethyl acetate. The combined organic phase was washed with 3 x50 mL of brine and dried over anhydrous sodium sulfate. The solution was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1:1). [(3R)-4-|2-(2-methoxyethyl)pyridine-3carbonyl]morpholin-3-yl]methanol was obtained. LCMS (ES) [M+l]+ m/z: 281.

Step 5

[0293] Into a 10-mL round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed [(2S)-l-[2-(2-niethoxyethyl)pyridme-3-carbonyl]pyrrolidin-2-yl]methanol 25 (280 mg, 1.06 mmol, 1.0 equiv), THF (10 mL), 2,6-dîhydroxybenzaldehyde (176 mg, 1.27 mmol, 1.2 equiv), and PPh3 (333 mg, 1.27 mmol, 1.2 equiv). The mixture was cooled to 0 °C followed by the addition of a solution of DBAD (293 mg, 1.27 mmol, 1.2 equiv) in THF (2 mL) dropwise with stirring. After addition, the reaction soluticn was stirred for 12 h at room température. The resulting mixture was concentrated in vacuum to remove the solvent, and the crude product was purified by Prep-HPLC with the following conditions (IntelFlash-1): Column: Ascentis Express Cl8, 50*3.0 mm, 2.7 um, Mobile Phase A: Water/0.05% FA, Mobile Phase B: CH3CN, Flow rate: 1.5 mL/min, Gradient: 5% B to 100% B within 1.2 min, hold 0.6 min. This

114 resulted ίη isolation of 2-hydroxy-6-[[(2S)-l-[2-(2-methoxyethyl)pyridine-3carbonyl]pyrrolidin-2-y]]methoxy]benzaldehyde. LCMS: (ES, m/z): [M+H]⁺: 401.2. Ή-NMR: (300 MHz, DMSO-d6, ppm): δ 11.76 (s, l H), 10.12 (s, 1H), 8.57 (dd, J - 4.8,1.8 Hz, 1H), 7.737.31 (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) pyrazine-2-carbonyl]morpholin-3yl]methoxy}benzaldehyde, Compound 21

[0294] Compound 21 was synthesized according to Scheme 17.

Scheme 17

[0295] Into a 100-mL 3-necked round-bottom flask, was placed 2-hydroxy-6-[[(3S)-4-[3-(2methoxyethyl)pyrazine-2-carbonyl]morpholin-3-yl]methoxy]benzaldehyde (500.00 mg, 1.25 mmol, 1.00 equiv), which was prepared as descrîbed in Scheme 23, and DCM (10.00 mL). This was followed by the addition of BBrfDCM (12.46 mL, 12.46 mmol, 10.00 equiv) dropwise with stirring at -78 °C. The resulting solution was stirred for 2 h at 0 °C. The resulting solution was extracted with 3x20 mL of dichloromethane, and the organic layers were combined, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-HPLC with the followîng conditions: Column, XBridge Prep C18 OBD Column, 19cm, 150mm, 5um; mobile phase, Water (0.1% HCOOH) and CAN (20% Phase B up to 50% in 11 min); Detector, 254. This resulted in 2-hydroxy-6-[[(3S)-4-[3-(2-hydroxyethyl)pyrazine-2-carbonyl]morpholin3-yl]methoxy]benzaldehyde. LCMS: (ES, m/z)'. [M+H] ⁺: 388. *H-NMR: (300 MHz, DMSOd₆)5 li.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), 3.51 - 3.30 (m, 2H), 3.17 - 2.80 (m, 3H).

115

Examplc 18. 2-hydiOxy-6-{[(3S)-4-[2-(2-hydroxyethyl)benzoyl]morpholin-3 yl]methoxy}benzaldehyde, Compound 22

[0296] Compound 22 was synthesized according to Scheme 18.

Scheme 18

Step 1

[0297] Into a 250-mL round-bottom flask, was placed niethyl 2-bromobenzoate (5.00 g, 23.251 mmol, 1.00 equiv), dioxane (60.00 mL), water (10 mL), 2-[(E)-2-ethoxyethenyl]-4,4,5,5tetramethyl-l,3,2-dioxaborolane (6.91 g, 34.876 mmol, 1.50 equiv), sodium methaneperoxoate sodium (4.98 g, 46.502 mmol, 2.00 equiv), and tetrakis(tiiphenylpbospbine)paIladium(0) (2.69 g, 2.325 mmol, 0.10 equiv). The resulting solution was stirred for 16 hr at 80 °C. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1:3) as eluents. The collected fractions were combined and concentrated. This resulted in methyl 2-[(E)-2-ethoxyethenyl]benzoate. LCMS (ES) [M+l]⁺ m/z

207.1.

116

Step 2

[0298] Into a 100-mL round-bottom flask, was placed rr ethyl 2-[(E)-2-ethoxyethenyl]benzoate (2.40 g, 11.637 mmol, 1.00 equiv), methanol (30.00 mL), and Pd/C (240.00 mg). The flask was evacuated and flushed three times with nitrogen, followec by flushîng with hydrogen. The mixture was stirred 2h at room température under an atmosphère of hydrogen. The solids were filtered out. The resulting filtrate was concentrated to give methyl 2-(2-ethoxyelhyl)benzoate. LCMS (ES) [M+l]⁺ m/z 209.1.

Step 3

[0299] Into a 50-mL round-bottom flask, was placed methyl 2-(2-ethoxyethyl)benzoate (2.00 10 g, 9.604 mmol, 1.00 equiv), methanol (10.00 mL), water (10.00 mL), caustic soda (0.77 g, 19.251 mmol, 2.00 equiv). The resulting solution was stirred for 4 hr at 25 °C. The resulting solution was diluted with 50 mL of water. The pH value of the solution was adjusted to 5 with HCl (1 mol/L). The solids were collected by filtration to give 2-(2-ethoxyethyl)benzoic acid. LCMS (ES) [M+l]⁺ m/z 195.1.

Step 4

[0300] Into a 100-mL round-bottom flask, was placed 2-(2-ethoxyethyl)benzoic acid (1.50 g, 7.723 mmol, 1.00 equiv), DCM (30.00 mL), (3R)-morpholin-3-ylmethanol (0.90 g, 7.723 mmol, 1.00 equiv), HATU (4.40 g, 11.584 mmol, 1.50 equiv), a .td D1EA (2.99 g, 23.168 mmol, 3.00 equiv). The resulting solution was stirred for 2 hr at 25 °C. The resulting mixture was concentrated. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1:1) as eluents. The collected fractions were combined and concentrated to give [(3R)-4-[2-(2ethoxyethyl)benzoyl]morpholin-3-yl]methanoI. LCMS (ES) [M+l]⁺ m/z 294.2.

Step 5

[0301] Into a 520-mL round-bottom flask, was placed [(3 R)-4-(2-(225 ethoxyethyl)benzoyl]morpholin-3-yl]methanol (600.00 mg, 2.045 mmol, 1.00 equiv), tetrahydrofuran (20 mL), 2,6-dihydroxybenzaldehyde (282.49 mg, 2.045 mmol, 1.00 equiv), triphenylphosphine (643.74 mg, 2.454 mmol, 1.20 equiv), and DIAD (496.28 mg, 2.454 mmol, 1.20 equiv). The resulting solution was stirred for 16 hr at 25°C. The resulting mixture was concentrated. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1:2) as eluents. The collected fractions were combined and concentrated. This resulted in 220663

117

[[(3S)-4-[2-(2-ethoxyethyl)benzoyl]morphoïm-3-yl]methoxy]-6-hydroxybenzaldehyde. LCMS (ES) [M+l]⁺ m/z 414.2.

Step 6

[0302] Into a 50-mL round-bottom flask, was placed 2-[[(3S)-4-[2-(25 ethoxyethyl)benzoyl]morpholin-3-yl]methoxy]-6-hydroxybenzaldehyde (200 mg, 0.48 mmol, 1.0 eq), DCM(20mL).Then, boron tribromide (2.4 mL, 2.4 mmol, 5.0 eq, IM) was added dropwise at -78°C. The resulting solution was stirred for 3 hr at 0°C. The reaction was then quenched by the addition of 10 mL of water. The resulting solution was extracted with 2x20 mL of dichloromethane and concentrated. The crude reaction mixture was filtered and subjected to reverse phase préparative HPLC (Prep-C18, 20-45M, 120 g, Tianjîn Bonna-Agela Technologies; gradient elution of 25% MeCN in water to 35% MeCN in water over a 10 min period, where both solvents contain 0.1% FA) to provide 2-hydroxy-6-{[(3S)-4-[2-(2hydroxyethyl)benzoyl]morpholm-3-yl]methoxy}bcnzaldehyde. LCMS (ES) [M+l]⁺ m/z 386.1. Ή NMR (300 MHz, DMSO-d6) δ I 1.76 (br, IH), 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).

118

Example 19. 2-hydroxy-6-{[(3S)-4-[2-(hydroxymethyI)benzoyl]morpholin-3 yl]methoxy}benzaldehyde, Compound 23

[0303] Compound 23 was synthesized according to Scheme 19.

Scheme 19

Step 1

[0304] Into a 500-mL round-bottom flask, was placed f hthalide (11.00 g, 82.008 mmol, 1.00 equiv), HîO (200.00 mL, 11101.675 mmol, 135.37 equiv), and NaOH (4.92 g, 123.009 mmol, 1.50 equiv). The resulting solution was stirred for overnight at 100 °C in an oil bath. The reaction mixture was cooled to 0 °C with a water/ice bath. The pH value of the solution was adjusted to 5 with HCI (6 mol/L). The solids were collected by filtration. The solid was dried in an oven. This resulted in 2-hy droxy methylbenzoic acid. LCMS (ES) [M-l]‘ m/z 151.1.

Step 2

[0305] Into a 250-mL round-bottom flask, was placed 2-hydroxymethylbenzoic acid (5.00 g, 15 32.863 mmol, 1.00 equiv), DCM (100.00 mL), and imidazole (4.47 g, 65.725 mmol, 2.00 equiv). This was followed by the addition of TBDPSC1 (10.84 g, 39.435 mmol, 1.20 equiv) in several batches at 0 °C in 5 min. The resulting solution was stirred for 2 h at room température. The resulting solution was diluted with 60 mL of water. The resulting solution was extracted with 3x100 mL of dichloromethane, and the organic layer was washed with 1x60 mL of brine,

119 dried over anhydrous sodium sulfate, and concentrated. T.re residue was purified by silica gel column chromatography with dichloromethane/elhyl acetate (10%EA-20%EA). This resulted in 2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoic acid. LCMS (ES) [M+l]⁺ m/z 391.2.

Step 3

[0306] Into a 250-mL round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed 2-[[(tert-butyldiphenylsîlyI)oxy]methyi]benzoic acid (2.00 g, 5.121 mmol, 1.00 equiv), DCM (120 mL), and oxalyl chloride (1.30 g, 10.242 mmol, 2.00 equiv). The resulting solution was stirred for 4 h at room température. The resulting mixture was concentrated under vacuum. Into another 250-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed (3R)-morpholîn-3-ylmethanol hydrochloride (0.94 g, 6.145 mmol, 1.20 equiv) and TEA (1.55 g, 15.318 mmol, 2.99 equiv). Thîs was followed by the addition of a solution of 2-[[(tei tbutyldiphenylsilyl)oxy]methyl]benzoyl chloride (2.00 g, 4.890 mmol, 1.00 equiv) in DCM (30 mL) dropwise with stirring at 0 °C in 30 min. The resulting solution was stirred for 2 h at room température. The resulting solution was diluted with 200 mL of DCM. The resulting mixture was washed with 1 x 100 mL of 1 M HCl. The mixture was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (50% EA). This resulted in [(3R)-4-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)morpholin-3-yl]methanol. LCMS (ES) [M+l]⁺ m/z

490.3.

Step 4

[0307] Into a 100-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed [(3R)-4-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)morpholin-3-yl]methanol (1.00 g, 2.042 mmol, 1.00 equiv), 2,6-dihydroxybenzaldehyde (0.56 g, 4.084 mmol, 2.00 equiv), PPh3 (1.07 g, 4.084 mmol, 2.00 equiv), and THF (60.00 mL). The resulting solution was stirred for 15 min at 0 ^ÜC. This was followed by the addition of DIAD (825.87 mg, 4.084 mmol, 2.00 equiv) dropwise with stirring at 0°C in 5 min. The resulting solution was stirred for 5 h at room température. The resulting solution was diluted with 30 mL of water. The resulting solution was extracted with

3x60 mL of dichloromethane; the organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography with PE/THF (10%THF). Thîs resulted in 2-[[(3S)-4-(2-[[(tert20663

120 butyldiphenylsilyl)oxy]methyl]benzoyl)morpholin-3-yl]methoxy]-6-hydroxybenzaldehyde. LCMS (ES) [M+l]⁺ m/z 610.3.

Step 5

[0308] Into a 100-mL round-bottom flask, was placed 2-[[(3S)-4-(2-[[(tert5 butyldiphenylsilyl)oxy]methyl]benzoyl)morpholin-3-yl]trethoxy]-6-hydroxybenzaldehyde (0.80 g, 1.312 mmol, 1.00 equiv), THF (30.00 mL), and TBAF (1.32 mL, 2.0 equiv, 2 M). The resulting solution was stirred for 2 hr at room température. The resulting solution was diluted with 10 mL of water. The resulting solution was extracted with 3x60 mL of dichloromethane, and the organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column with ΡΕ/THF (55%THF) as eluents. The crude product was further purified by Flash-Prep-HPLC (Prep-C18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies; gradient elution of 35% MeCN in water to 60% MeCN in water over a 10 min period, where both solvents contain 0.1% FA). This resuked in 2-hydroxy-6-[[(3S)-4-[2(hydroxymethyl)benzoyl]morpholin-3-yl]methoxy]benzaldehyde. LCMS (ES) [M+l]⁺ m/z

372.1. ’H-NMR (300 MHz, DMSO-ri^ppm) δ 11.78 (s, 1 H), 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), 4.183.84 (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)-l-[2-(2-methoxyethyl ipyridine-3-carbonyl]pyrrolidin-2yl]methoxy}benzaldehyde, Compound 24

[0309] Compound 24 was synthesized according to Scheme 20.

Scheme 20

120 butyldiphenylsilyl)oxy]methyl]benzoyl)morpholin-3-yl]methoxy]-6-hydroxybenzaldehyde.

LCMS (ES) [M+l ]⁺ m/z 610.3.

Step 5

[0308] Into a 100-mL round-bottom flask, was placed 2-[[(3S)-4-(2-[[(tert5 butyldiphenylsi[yl)oxy]methyl]benzoyl)morpholin-3-yl]methoxy]-6-hydroxybenzaldehyde (0.80 g, 1.312 mmol, 1.00 equiv), THF (30.00 mL), and TBAF (1.32 mL, 2.0 equiv, 2 M). The resulting solution was stirred for 2 hr at room température. The resulting solution was diluted with 10 mL of water. The resulting solution was extracted with 3x60 mL of dichloromethane, and the organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column with ΡΕ/THF (55%THF) as eluents. The crude product was further purified by Flash-Prep-HPLC (Prep-C18, 20-45M, 120 g, Tianjîn Bonna-Agela Technologies; gradient elution of 35% MeCN in water to 60% MeCN in water over a 10 min period, where both solvents contain 0.1% FA). This resul.ed in 2-hydroxy-6-[[(3S)-4-[2(hydroxymethyl)benzoyl]morpholin-3-yI]methoxy]benzaldehyde. LCMS (ES) [M+l]⁺ m/z

372.1. *H-NMR (300 MHz, DMSO-d_6ippm) δ 11.78 (s, 1 H), 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), 4.183.84 (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)-l-[2-(2-methoxyethyl)pyridine-3-carbonyl]pyrrolidin-2yl]methoxy}benzaldehyde, Compound 24

[0309] Compound 24 was synthesized according to Scheme 20.

Scheme 20

121

Step 1

[0310] Into a 50-mL 3-necked round-bottom flask purgcd and maintained with an inert atmosphère of nitrogen, was placed 2-(2-methoxyethyl)pyridine-3-carboxylic acid (1.0 g, 5.52 mmol, 1.0 equiv), DMF (20 mL), prolinol (670 mg, 6.62 mmol, 1.2 equiv), and DIEA (2.85 g, 5 22.08 mmol, 4.0 equiv). This was followed by the addition of HATU (2.52 g, 6.62 mmol, 1.2 equiv) in several batches at 0 °C. After addition, the mixture was stirred for 12 h at room température. The reaction solution was diluted with 30 mL of HzO and extracted with 3x100 mL of ethyl acetate. The combined organic phase was washec with 3*50 mL of brine and dried over anhydrous sodium sulfate. The solution was filtered, and :he filtrate was concentrated under reduced pressure. The residue was purified by silica gel column with ethyl acetate. [(2S)-l-[2(2-methoxyethyl)pyridine-3-carbonyl]pyrrolidin-2-y]]melhanol was obtained. LCMS (ES) [M+l]⁺ m/z: 265.

Step 2

[0311] Into a 50-mL round-bottom flask purged and maintained with an inert atmosphère of 15 nitrogen, was placed [(2S)-l-[2-(2-methoxyethyl)pyridine-3-carbonyl]pyrrolidin-2-yl]methanol (380 mg, 1.44 mmol, 1.0 equiv), THF (20 mL), 2,6-dihyclroxybenzaldehyde (199 mg, 1.44 mmol, 1.0 equiv), and PPh3 (377 mg, 1.44 mmol, 1.0 equiv). The mixture was cooled to 0 ^ÛC and stirred for 15 min. This was followed by the addition of a solution of DBAD (331 mg, 1.44 mmol, 1.0 equiv) in THF (2 mL) dropwise with stirring. .After addition, the réaction solution was 20 stirred for 12 h at room température. The solution was then concentrated under reduced pressure to remove the solvent. The residue was purified by Flash-Prep-HPLC with the following conditions (IntelFlash-1): Column: Ascentîs Express C18, 50*3.0 mm, 2.7 um, Mobile Phase A: Water/0.05% FA, Mobile Phase B: CH3CN, Flow rate: 1 5 mL/min, Gradient: 5% B to 100% B within 1.2 min, hold 0.6 min. 2-hydroxy-6-{[(2S)-l-[2-(2-methoxyethyI)pyridine-325 carbony 1]pyrrolidin-2-yl]methoxy}benzaldehyde was obtained. LCMS; (ES, m/z): [M+H]⁺: 385.

’H-NMR: (300 MHz, DMSO-d_c„ ρρηϊ): 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), 3.793.52 (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).

122

Example 21. 2-hydroxy-6-{[(2S)-l-[3-(2-methoxyethyl)pyrazine-2-carbonyl]pyrrolidin-2y l]methoxy [benzaldehyde, Compound 25

[0312] Compound 25 was synthesized according to Scheme 21.

Step 1

[0313] Into a 250-mL round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed methyl 3-bromopyrazîne-2-carboxyÎate (5.00 g, 23.04 mmol, 1.00 equiv), 2-ethenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (4.26 g, 27.66 mmol, 1.20 equiv), dioxane (60.00 mL), H₂O (10.00 mL), K₂CO₃ (6.37 g, 46.08 mmol, 2.00 equiv), and Pd(dppf)Cl₂ (1.69 g,

2.30 mmol, 0.10 equiv). The resulting solution was stirred for 5 h at 80 °C. The reaction mixture was cooled to room température. The solids were filtered out. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with THF/PE (15%) as eluents. This resulted in methyl 3-ethenylpyrazine-2-carboxylate. LCMS (ES) [M+l]⁺ 15 m/z: 165.

Step 2

[0314] Into a 250-mL round-bottom flask, was placed methyl 3-ethenylpyrazine-2-carboxylate (3.50 g, 21.32 mmol, 1.00 equiv), MeOH (40.00 mL), and NaOMe (3.46 g, 64.05 mmol, 3.00

123 equiv), The resulting solution was stirred for overnight at 70 °C. The resulting mixture was concentrated. The pH value of the solution was adjusted to 2-3 with HCl (1 mol/L). The resulting mixture was concentrated, The residue was purified by silica gel column with MeOH/DCM (10%) as eluents. This resulted in 3-(2-metboxyethyI)pyrazine-2-carboxyIic acid, 5 LCMS (ES) [M+l]⁺ m/z: 183.

Step 3

[0315] Into a 250-mL 3-necked round-bottom flask, was placed 3-(2-methoxyethyl)pyrazine2-carboxylic acid (1.50 g, 8.23 mmol, 1.00 equiv), prolinol (0.83 g, 8.21 mmol, 1.00 equiv), DIEA (3.19 g, 24.70 mmol, 3.00 equiv), and DMF (30.00 mL). This was followed by the addition of HATU (3.76 g, 9.88 mmol, 1.20 equiv) in portons at 0 °C. The resulting solution was stirred for overnight at room température. The crude oroduct was purified by Prep-HPLC with the following conditions: Column, XBridge Prep Cl 3 OBD Column, 19cm, 150mm, 5um; mobile phase, Water (0.1% HCOOH) and MeCN (5% Phase B up to 20% in 10min); Detector, 254. This resulted in [(2S)-l-[3-(2-methoxyethyl)pyrazine-2-carbonyl]pyrrolidin-2-yl]methanol.

LCMS (ES) [M+l]⁺ m/z: 266.

Step 4

[0316] Into a 100-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed [(2S)-l-[3-(2-methoxyethyl)pyrazine-2-carbonyl]pyrrolidin2-yl]methanol (1.00 g, 3.77 mmol, 1.00 equiv), 2,6-dihydroxybenzaldehyde (0.62 g, 4.49 mmol, 20 1.19 equiv), PPh.3 (1.19 g, 4.52 mmol, 1.20 equiv), and DCM (30.00 mL). This was followed by the addition of DIAD (0.91 g, 4.52 mmol, 1.20 equiv) dropwise with stirring at 0 °C. The resulting solution was stirred for overnight at room température. The resulting mixture was concentrated. The residue was applied onto a silica gel column with EA/DCM (10%). This resulted in 2-hydroxy-6-[[(2S)-l-[3-(2-methoxyethyl)pyrazine-2-carbonyI]pyrrolidin-225 yl]methoxy]benzaldehyde. LCMS: (ES, m/z): [M+H] ⁺: 386.2. *H-NMR: (300 MHz, DMSOdè) ÔÔ 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).

124

Example 22. 2-hydroxy-6-{[(2S)-l-[2-(hy droxy methyl) benzoyl]pyrro lidin-2yl]methoxy}benzaldehyde, Compound 26

[0317] Compound 26 was synthesized according to Scheme 22.

Step 1

[0318] Into a 100-mL round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed 2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoic acid (2.00 g, 5.121 mmol, 1.00 equiv), DCM (60.00 mL), and DMF (0.05 mL, 0.646 mmol, 0.13 equiv). This was followed 10 by the addition of oxalyl chloride (1.30 g, 10.243 mmol, 2.00 equiv). The resulting solution was stirred for 1 overnight at room température. The resulting mixture was concentrated. This resulted in 2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl chloride.

Step 2

[0319] Into a 250-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed prolinol (0.59 g, 5.86'8 mmol, 1.2 equiv), TEA (1.48 g, 14.670 mmol, 3 equiv), and DCM (100.00 mL). This was followed by the addition of a solution of 2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl chloride (2.00 g, 4.890 mmol, 1.00 equiv) in DCM (30 mL) dropwise with stirring at 0°C in 30 min. The resulting solution was stirred for 2 h at room température. The resulting solution was diluted with 100 mL of DCM. The resulting

125 mixture was washed with 1 x 70 mL of 1 M HCl. The mbture was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (55%EA) as eluents. This resulted in [(2S)-l-(2-[[(tertbutyldiphenylsilyi)oxy]methyl]benzoyl)pyrrolidin-2-yl]methanol. LCMS (ES) [M+l]⁺ m/z 5 474.2.

Step 3

[0320] Into a 100-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed [(2S)-l-(2-[[(tertbutyldiphenylsîlyl)oxy]melhyl]benzoyI)pyiTolidin-2-yl]methanol (1.00 g, 2.111 mmol, 1.00 equiv), 2,6-dihydroxybenzaldehyde (0.58 g, 4.222 mmol, 2.00 equiv), PPh₃ (1.11 g, 4.222 mmol, 2.00 equiv), and THF (60 mL). The resulting solution was stirred for 15 min at 0 °C. This was followed by the addition of DIAD (0.85 g, 4.222 mmol, 2.00 equiv) dropwise with stirring at 0 °C in 5 min. The resulting solution was stirred for 5 h at room température. The resulting solution was diluted with 30 mL of water. The resulting solution was extracted with 3x100 mL of dichloromethane; the organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography with PE/THF (12%THF) as eluents. This resulted in 2-[[(2S)-l-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)pyrrolidin-2-yl]ir.ethoxy]-6-hydroxybenzaldehyde. LCMS (ES) [M+l]⁺ m/z 594.3.

Step 4

[0321] Into a 100-mL round-bottom flask, was placed 2-[[(2S)-l-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)pyrrolîdin-2-yl]niethoxy]-6-hydroxybenzaldehyde (1.10 g, 1.852 mmol, 1.00 equiv), THF (30.00 mL), and TBAF (1.9 mL, 2 M). The resulting solution was stirred for 2 h at room température. The resulting solution was diluted with 10 mL of water.

The resulting solution was extracted with 3x60 mL of dichloromethane; the organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was applied onto a silica gel column with PE/THF (52%THF). The crude product was purified by Flash-Prep-HPLC (PrepC18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies; gradient elution of 35% MeCN in water to 60% MeCN in water over a 10 min period, where both solvents contain 0.1% FA). This resulted in 2-hydroxy-6-[[(2S)-l-[2-(hydroxymethyl)benzoyl]pyrrolidin-2yl]methoxy]benzaldehyde. LCMS (ES) [M+l]⁺ m/z 356.1. Ή-NMR (300 MHz, DMSO-^ppm)

126 ’ δ I 1.72 (s, 1Η), 10.22 (s, 1H), 7.80-7.13 (m, 5H), 6.78-6.41 (m, 2H), 5.22-5.10 (m, 1H), 4.584.28 (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-hydroxy-6-{[(3S)-4-[3-(2-methoxyethyl)pyrazine-2-carbonyl]morpholin-3yl]methoxy}benzaldehyde, Compound 27

[0322] Compound 27 was synthesized according to Scheme 23.

Scheme 23

Step 1

[0323] Into a 100-mL 3-necked round-bottom flask, was placed 3-(2-methoxyethyl)pyrazine10 2-carboxylic acid (2.00 g, 10.98 mmol, 1.00 equiv), (3S)-3-[[(tertbutyldimethylsilyl)oxy]methyl]morpholine (2.54 g, 10.98 mmol, 1.00 equiv), EcN (2.22 g, 21.94 mmol, 2.00 equiv), DCM (30 mL), and EDCI (2.53 g, 13.17 mmol, 1.20 equiv). This was followed by the addition of HOBt (1.78 g, 13.17 mmol, 1.20 equiv) in portions at 0 °C. The resulting solution was stirred for overnight at room température. The reaction was then quenched 15 by the addition of 30 mL of water. The resulting solution was extracted with 3x30 mL of dichloromethane, and the organic layers were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column with THF/PE (40%) as eluents. This resulted in (3S)-3-[[(tert-butyldimethylsilyl)oxy]methyl]-4-[3-(2-methoxyethyl)pyrazine-2carbonyi]morpholine. LCMS (ES) [M+l]⁺ m/z: 396.

127

Step 2

[0324] Into a 100-mL round-bottom flask, was placed (3 S)-3-[[(tertbutyldimethylsilyl)oxy]methyl]-4-[3-(2-methoxyethyl)pytazine-2-carbonyl]morpholine (4 g, 10.11 mmol, 1.00 equiv) and EA (20.00 mL). To the above HCl(_g) in EA (10.11 mL, 20.22 mmol, 2.00 equiv) was introduced in dropwîse with stirrirg at 0 °C. The resulting solution was stirred for 2 h at room température. The resulting mixture was concentrated. The pH value of the solution was adjusted to 7-8 with saturated NaHCOs. The resulting solution was extracted with 5x30 mL of dichloromethane, and the organic layers were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column with dîchloromethane/methanol (100/3) as eluents. This resulte d in [(3R)-4-[3-(2methoxyethyl)pyrazine-2-carbonyl]morpholin-3-yl]methanol. LCMS (ES) [M+l]⁺ m/z: 282.

Step 3

[0325] Into a 100-mL 3-necked round-bottom flask, was placed [(3R)-4-[3-(2methoxyethyl)pyrazine-2-carbonyl]morpholin-3-yl]methanol (400.00 mg, 1.42 mmol, 1.00 equiv), 2,6-dihydroxybenzaldehyde (235.68 mg, 1.71 mmol, 1.20 equiv), DCM (10.00 mL), and PPM (447.54 mg, 1.71 mmol, 1.20 equiv). This was followed by the addition of DIAD (345.03 mg, 1.71 mmol, 1.20 equiv) dropwîse with stirring at 0 °C. The resulting solution was stirred for overnight at room température. The resulting mixture was concentrated. The crude product was purified by Prep-HPLC with the following conditions: Column, XBridge Prep C18 OBD

Column, 19cm, 150mm, 5um; mobile phase, Water (0.1% HCOOH) and CAN (30% Phase B up to 50% in 11 min); Detector, 254. This resulted in 2-hydroxy-6-{[(3S)-4-[3-(2methoxyethyl)pyrazine-2-carbonyl]morpholin-3-yl]methoxy}benzaldehyde. LCMS: (ES, m/z): [M+H] ⁺: 402. ’H-NMR: (300 MHz, DMSO-d_t.) 5 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), 6.64 - 6.48 (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).

128

Example 24. 3-{3-[(3S) -3-[(2-fonnyl -3-hydroxyphenoxy) methyl] morpholine-4carbonyl]pyridin-2-yl}propanenitrile, Compound 28

[0326] Compound 28 was synthesized according to Scheme 24.

Scheme 24

24a

NC^

Step 1

Step 2

Step 4

Step 1

[0327] Into a 40-mL vial was placed methyl 2-cbloropyridine-3-carboxylale (2.00 g, 11.66 mmol, 1.00 equiv), DMF (15.00 mL), NaOAc (1.91 g, 23.31 mmol, 2.00 equiv), PPh₃ (1.22 g, 4.66 mmol, 0.40 equiv), Pd(OAc)₂ (0.26 g, 1.17 mmol, 0.10 equiv), and acrylonitrile (3.09 g, 10 58.28 mmol, 5.00 equiv). The resulting solution was stirrïd for overnîght at 130 °C in an oil bath. The reaction was then quenched by water (20 mL) and extracted with EA (40mL). The organic layer was washed by water (40 mL) two times, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (0-9.9%) as eluents. This resulted in methyl 2-[(lE)-2-cyanoeth-l15 en-l-yl]pyridine-3-carboxyiate. LCMS: (ES, m/z)·. [Μ+Η]⁺ 189.1.

I

129

Step 2

[0328] Into Into a 50-mL round-bottom flask, was placed methyl (E)-2-(2cyanovinyl)nicotinate (1.4Ü g, 7.44 mmol, 1.00 equiv), CHjOH (20 mL), and Pd/C (140.0 mg, 10%). To the above, H2 was introduced. The resulting solution was stirred for 1 hr at room 5 température. The solids were filtered oui, and the mixture was concentrated and purified by silica gel column chromatography with ethyl acetate/petrcleum ether (0-15%) as eluents. This resulted in methyl 2-(2-cyanoethyl)nicotinate. LCMS: (ES, m/z): [M+H]⁺: 190.1.

Step 3

[0329] Into a 50-mL round-bottom flask, was placed methyl 2-(2-cyanoethyl)nicotinate (1.00 10 g, 5.26 mmol, 1.00 equiv) and THF (12 mL). Then, a solution of LiOH (0.44 g, 10.49 mmol,

1.99 equiv) in H₂O (6mL) was added. The resulting solution was stirred for 1 hr at room température and concentrated. The pH value of the solution was adjusted to 5-6 with HCl (2 mol/L). The residue was purified with the following conditions: column, Cl 8; mobile phase, water (0.05% FA) and CH3CN (5% up to 80% in 8 min); Detector, 220 & 254nm; Flow rate, 15 40mL/min. This resulted in 2-(2-cyanoeth-yl)nicotÎnic acid. LCMS (ES, m/z): [M+H] ⁺: 177.1.

Step 4

[0330] Into a 50-mL round-bottom flask, was placed 2-(2-cyanoethyl)pyridine-3-carboxylic acîd (300.0 mg, 1.70 mmol, 1.00 equiv), HATU (777.0 mg, 2.04 mmol, 1.20 equiv), DMF (10 mL), DIEA (550.2 mg, 4.26 mmol, 2.50 equiv), and (3S)-3-[[(tert20 butyldimethylsilyl)oxy]methyl]morpholine cyclohexane (430.0 mg, 1.86 mmol, 1.09 equiv). The resulting solution was stirred for 2 hr at room température. The reaction was then quenched by the addition of water. The resulting solution was extracted with 3x20 mL of ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (0-50%) as eluents. This resulted in (S)-3-(325 (3-(((tert-butyldimethyIsilyl)oxy)methyl)morpholine-4-carbonyl)pyridin-2-yl)-propanenitrile.

LCMS (ES, m/z): [M+H] ⁺: 390.2.

Step 5

[0331] Into a 50-mL round-bottom flask, was placed (S)-3-(3-(3-(((tertbutyldimethylsîlyl)oxy)methyl)-morpholine-4-carbonyl)pyridin-2-yl)-propanenitrile (0.63 g, 30 1.62 mmol, 1.00 equiv), THF (10 mL, 123.43 mmol, 76.3 equiv), and TBAF(i.O M) (2.43 mL,

2.43 mmol, 1.50 equiv). The resulting solution was stirred for 2 hr at room température. The

130 resulting mixture was concentrated and purified by silica gel column chromatography with dichloromethane/methanol (94.6:5.4) as eluents. This resulted in (R)-3-(3-(3(hydroxymethyl)morpholine-4-carbonyl)pyridin-2-yl)propanenitrile. LCMS (ES, m/z): [M+H] ⁺: 276.1.

Step 6

[0332] Into a 40-mL vial purged and maintaîned with ar inert atmosphère of nitrogen, was placed (R)-3-(3-(3-(hydroxymethyl)morpholine-4-carbonyl)pyridin-2~yl)piOpanenitrile (0.24 g, 0.87 mmol, 1.00 equiv), THF (10 mL), PPh₃ (274.4 mg, 1.05 mmol, 1.20 equiv), and 2,6dihydroxybenzaldehyde (156.5 mg, 1.13 mmol, 1.30 equiv). Then, DBAD (240.9 mg, 1.05 mmol, 1.20 equiv) was dropwise at 0 °C. After 20 min, the resulting solution was stirred at 40 °C for overnight. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (0-80%) as eluents. The crude product was purified by Prep-HPLC with the followîng conditions; Column, Kinetex EVO Cl8, 21.2* 150mm, 5 um; Mobile phase; water(0.1% FA) and CH₃CN( 35% up to 75% in 14min).

Detector; 220nm. Flow rate, 20mL/min. This resulted in 3-{3-[(3S)-3-[(2-formyl-3hydroxyphenoxy)methyl] morpholine-4-carbonyl]pyridin-2-yl}propanenitrile. LCMS: (ES, m/z): [M+H] ⁺: 396.2. Ή-ΝΜ: (300 MHz, DMSO-î/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), 4.944.89 (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-{[(3S)-4-[2-(2-methoxyethyl)benzoyl]morpholin-3yl]methoxy}benzaldehyde, Compound 29

[0333] Compound 29 was synthesîzed according to Scheme 25.

Scheme 25

131 r Step 1

[0334] Into a 100-mL round-bottom flask, was placed 2 -(2-methoxyethyl)benzoic acid (500. ÜO mg, 2.775 mmol, 1.00 equiv), DCM (20.00 mL), (3R)-mcrpholin-3-ylmethanol (325.04 mg, 2.775 mmol, 1.00 equiv), HATU (1582.51 mg, 4.162 mmol, 1.50 equiv), and DIEA (1075.81 mg, 8.324 mmol, 3.00 equiv). The resulting solution was stirred for 3 hr at 25 °C. The resulting mixture was concentrated. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1:1) as eluents. The collected fractions were combîned and concentrated. This resulted in [(3R)-4-[2-(2-methoxyethy .)benzoyI]morpholin-3-yl]methanol. LCMS (ES) [M+l]⁺ m/z 280.2.

Step 2

[0335] Into a 50-mL round-bottom flask, was placed [(3R)-4-[2-(2methoxyethyl)benzoyi]morpholin-3-yl]methanol (200.00 mg, 0.716 mmol, 1.00 equiv), tetrahydrofuran (10 mL), 2,6-dihydroxybenzaldehyde (98.89 mg, 0.716 mmol, 1.00 equiv), triphenylphosphine (225.36 mg, 0.859 mmol, 1.20 equiv), and DIAD (173.73 mg, 0.859 mmol, 15 1.20 equiv). The resulting solution was stirred for 16 hr ai: 25 C. The resulting mixture was concentrated. The crude reaction mixture was filtered and subjected to reverse phase préparative HPLC (Prep-C18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies; gradient elution of 25% MeCN în water to 35% MeCN in water over a 10 min period, where both solvents contain 0.1% FA) to provide 2-hydroxy-6-{[(3S)-4-[2-(2-methoxyethyl)benzoyl]morpholin-320 yl]melhoxy}benzaldehyde. LCMS (ES) [M+lf m/z 400.2. Ή NMR (300 MHz, DMSO-d6) δ 11.76 (br, 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).

132

Example 26. 2-hydroxy-6-([(3S)-4-[2-(2-methoxyethyl)benzoyl]morphoiin-3yl]methoxy}benzaldeliyde, Compound 30

[0336] Compound 30 was synthesized according to Scheme 26.

Scheme 26

26c

Step 1

[0337] Into a 50-mL round-bottom flask, was placed 2-( 2-cyanoethyl)pyridine-3-carboxylic acid (0.30 g, 1.70 mmol, 1.00 equiv), HATU (777.0 mg, 2.04 mmol, 1.20 equiv), DMF (10.0 mL), DIEA (550.2 mg, 4.26 mmol, 2.50 equiv), and (2S)-2-[[(tert10 butyldimethylsilyl)oxy]methyl]pyrrolidine (403.50 mg, 1.87 mmol, 1.10 equiv). The resulting solution was stirred for 2 hr at room température. The reaction was then quenched by the addition of water (20mL), extracted with 3 x 20 mL of ethyl acetate, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatogrpahy with ethyl acetate/petroleum ether (0-60%) as eluents. This resulted (S)-3-(3-(2-(((tert15 butyldimethylsilyl)oxy)methyl)pyrrolidine-l-carbonyl)pyridin-2-yl)propanenitrile. LCMS (ES, m/z)i [M+H] ⁺: 374.2.

Step 2

[0338] Into a 50-mL round-bottom flask, was placed (S)-3-(3-(2-(((tertbutyldimethylsi!yl)oxy)methyl)-pyrrolidine-l-carbonyl)pyridin-2-yl)propanenitrile (0.4 g, 1.07

I

133 mmol, l.Ü equiv), THF (10 mL), and TBAF (1.2 mL, 1.2C mmol, 1.1 equiv). The reaction solution was stirred for 2 hr at room température. The resulting mixture was concentrated and purified by silica gel column chromatography with ethyl MeOH/DCM (6:94) as eluents.

This resulted in (S)-3-(3-(2-(hydroxymethyl)pyrrolidine-1 -carbonyl)pyridin-2-yl)propanenitrile.

LCMS (ES, m/z): [M+H] ⁺: 260.1.

Step 3

I

[0339] Into a 40-mL vial purged and maintained with an inert atmosphère of nitrogen, was placed (S)-3-(3-(2-(hydroxymethyl)pyrrolidine-1 -carbonyl)pyridin-2-yl)propanenitrile (120.00 mg, 0.46 mmol, 1.00 equiv), PPhi (145.6 mg, 0.56 mmol, 1.20 equiv), THF (10 mL), and 1-(2,610 dihydroxyphenyl)ethanone (91.5 mg, 0.60 mmol, 1.30 equiv). Then, DBAD (127.9 mg, 0.56 mmol, 1.20 equiv) was dropwise at 0 °C. After 20 min, the resulting solution was stirred at 40 °C for overnight. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (0-90%) as eluents. The crude product was purified by Prep-HPLC with the following conditions: Column, Kinetex EVO Cl S,

21.2*150mm, 5 uni; Mobile phase water (0.1% FA) and CHrCN (40% up to 70% in 14min);

Detector, 220nm. Flow rate, 20mL/min. This resulted in %hydroxy-6-{[(3S)-4-[2-(2methoxyethyl)benzoyl]morpholin-3-yl]methoxy}benzaîdehyde. LCMS: (ES, m/z): [M+H] ⁺: ।

380.2 . ’H-NMR (300 MHz, DMSO-î/6, ppm): δ 1 1.68 (s, 1 H), 10.34 (s, 1 H), 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).

134

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

[0340] Compound 31 was synthesized according to Scheme 27.

Scheme 27

Step 1

[0341] Into a 100-mL 3-necked round-bottom flask purged and maîntained with an inert atmosphère of nitrogen, was placed methyl 2-bromobenzoate (3.00 g, 13.951 mmol, 1.00 equiv), acrylonitrile (1.48 g, 27.901 mmol, 2.00 equiv), DIEA (5.41 g, 41.859 mmol, 3.00 equiv), dioxane (50.00 mL), and Pd(P(t-Bu)₃)₂ (0.71 g, 1.389 mmol, 0.10 equiv). The resulting solution was stirred for 16 h at 100 °C in an oil bath. The resulting mixture was concentrated. The residue was purified by silica gel column with ethyl acetate/petrcleum ether (10%EA) as eluents. This resulted in methyl 2-[(lE)-2-cyanoeth-l-en-l-yl]benzoate. LCMS (ES) [M+l] m/z 188.0.

Step 2

[0342] Into a 100-mL round-bottom flask, was placed methyl 2-[(lE)-2-cyanoeth-l-en-lyl]benzoate (1.90 g, 10.150 mmol, 1.00 equiv), MeOH (40.00 mL, 987.956 mmol, 97.34 equiv),

135 ) and Pd/C (0.80 g, 7.517 mmol, 0.74 equiv). The flask was evacuated and flushed three times with nitrogen, followed by flushing with hydrogen. The mixture was stirred 4 h at room température under an atmosphère of hydrogen (balloon). The solids were filtered out. The fîltrate was concentrated. This resulted in methyl 2-(2-cyanoethyl)benzoate. LCMS (ES) [M+l]⁺ 5 m/z 190.1.

Step 3

L0343] Into a 100-mL round-bottom flask, was placed methyl 2-(2-cyanoethyl)benzoate (1.90 g, 10.042 mmol, 1.00 equiv) andMeOH (50.00 mL). This was followed by the addition of a solution of LiOH (0.72 g, 30.065 mmol, 2.99 equiv) in H2O (10 mL) at 0 °C in 5 min. The resulting solution was stirred for 16 h at room température. The pH value of the solution was adjusted to 5 with citric acid (3 mol/L). The resulting solution was extracted with 3x100 mL of DCM/MeOH=10:l. The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. This resulted in 2-(2-cyanoethyl)benzoic acid. LCMS (ES) [M-l]' m/z 174.3.

Step 4

[0344] Into a 20-mL sealed tube, was placed 2-(2-cyanoethyl)benzoic acid (0.50 g, 2.854 mmol, 1.00 equiv), (3R)-morphoIin-3-ylmethanol hydrochloride (0.66 g, 4.281 mmol, 1.50 equiv), HATU (1.63 g, 4.281 mmol, 1.50 equiv), DIEA (1.11 g, 8.588 mmol, 3.01 equiv), and DMF (10.00 mL). The resulting solution was stirred for 2 h at room température. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with

PE:THF (45%THF) as eluents. This resulted in 3-[2-[(3R)-3-(hydroxymethyl)morpholine-4carbonyl]phenyl]propanenitrile. LCMS (ES) [M+l]⁺ m/z 275.1.

Step 5

[0345] Into a 100-mL round-bottom flask purged and maintained with an înert atmosphère of nitrogen, was placed 3-[2-[(3R)-3-(hydroxymethyl)morpholine-425 carbonyl]phenyl]propanenitrile (0.40 g, 1.458 mmol, 1.00 equiv), 2,6-dihydroxybenzaldehyde (0.40 g, 2.916 mmol, 2 equiv), PPh₃ (0.76 g, 2.916 mmol, 2 equiv), and THF (30.00 mL). The resulting solution was stirred for 15 min at 0 °C. This was followed by the addition of DIAD (0.59 g, 2.918 mmol, 2.00 equiv) at 0 °C in 3 min. The resulting solution was stirred for 2 h at room température. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with ΡΕ/THF (22%THF) as eluents. The collected product was purified by Flash-Prep-HPLC (Prep-C18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies;

136 gradient elution of 45% MeCN in water to 65% MeCN in water over a 10 min period, where both solvents contain 0.1% FA). This resulted in 3-{2-[(3S)-3-[(2-formyl-3hydroxyphenoxy)methyl]morpholine-4-carbonyl]phenyl}propanenitrile. LCMS (ES) [M+l]⁺ m/z 395.2. Ή NMR (300 MHz, DMSO-d^^m) δ 11.73 (s, II I), 10.28 (s, 1H), 7.63-7.03 (m, 5 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]morphoIine-4 carbonyl]pyrazin-2-yl}propanenitrile, Compound 32

[0346] Compound 32 was synthesized according to Scheme 28.

Scheme 28

28a 28b 28c

Step 1

[0347] Into a 100-mL 3-necked round-bottom flask purgcd and maintained with an inert atmosphère of nitrogen, was placed methyl 3-bromopyrazine-2-carboxylate (6.00 g, 27.647 mmol, 1.00 equiv), acrylonitrile (4.40 g, 82.941 mmol, 3 equiv), D1EA (10.72 g, 82.941 mmol, 3 equiv), dioxane (60.00 mL), and Pd(P(t-Bu)a)2 (1.41 g, 2.765 mmol, 0.1 equiv). The resulting

137 solution was stirred for 16 h at 100 °C in an oil bath. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with (10%-20%EA) as eluents. This resulted in methyl 3-[(lE)-2-cyanoeth-l-en-l-yl]pyrazine-2-carboxylate. LCMS (ES) [M+l]⁺ m/z 190.2.

Step 2

[0348] Into a 100-mL round-bottom flask, was placed methyl 3-[(lE)-2-cyanoeth-l-en-lyljpyrazine-2-carboxylate (1.60 g, 8.458 mmol, 1.00 equiv), MeOH (20.00 mL), and Pd/C (0.60 g, 5.638 mmol, 0.67 equiv). The flask was evacuated and flushed three times with nitrogen, foliowed by flushing with hydrogen. The resulting solution was stirred for 4 h at room température under an atmosphère of hydrogen (balloon). The solids were filtered out. The filtrate was concentrated. This resulted in methyl 3-(2-cyanoethyl)pyrazine-2-carboxylate. LCMS (ES) [M+l]⁺ m/z 192.2.

Step 3

[0349] Into a 100-mL round-bottom flask, was placed methyl 3-(2-cyanoethyl)pyrazine-215 carboxylate (0.70 g, 3.661 mmol, 1.00 equiv) and MeOH (50 mL). This was foliowed by the addition of a solution of LiOHH₂O (0.31 g, 7.387 mmol, 2.02 equiv) in H₂O (10 mL) dropwise with stirring at 0 °C in 5 min. The resulting solution was stirred for 2 h at room température.

The pH value of the solution was adjusted to 5 with citric acid (2 mol/L). The resulting solution was extracted with 20x100 mL of DCM/MeOH=10: l, and the organic layer was concentrated.

This resulted in 3-(2-cyanoethyl)pyrazine-2-carboxylîc acid. LCMS (ES) [M-l]’ m/z 176.1.

Step 4

[0350] Into a 20-mL vial, was placed 3-(2-cyanoethyl)pyrazine-2-carboxylic acid (0.43 g, 2.427 mmol, 1.00 equiv), (3R)-morpholin-3-ylmethanol hydrochloride(0.34 g, 2.913 mmol, 1.2 equiv), HATU (1.11 g, 2.913 mmol, 1.20 equiv), DIEA (0.94 g, 7.281 mmol, 3.00 equiv), and 25 DMF (10.00 mL). The resulting solution was stirred for 2 h at room température. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with ΡΕ/THF (50%THF) as eluents. This resulted in 3-[3-[(3R)-3-(hydroxymethyl)morpholine-4carbonyl]pyrazin-2-yl]propanenitrile. LCMS (ES) [M+l]⁺ m/z 277.1.

Step 5

138 ) [0351] Into a lüO-mL 3-necked round-bottom flask purged and maîntaîned with an inert atmosphère of nitrogen, was placed 3-[3-[(3R)-3-(hydroxymethyl)morpholine-4carbonyl]pyrazin-2-yl]propanenitrile (200.00 mg, 0.724 mmol, 1.00 equiv), 2,6dihydroxybenzaldehyde (199.96 mg, 1.448 mmol, 2.00 equiv), PPhs (379.72 mg, 1.448 mmol, 5 2.00 equiv), and THF (20.00 mL). The resulting solution was stirred for 15 min at 0°C. This was followed by the addition of DIAD (292.74 mg, 1.448 mmol, 2.00 equiv) dropwise with stirrîng at 0 °C în 5 min. The resulting solution was stirred for 2 h at room température. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with ΡΕ/THF (35%THF) as eluents. The coliected product was further purified by Flash-Prep-HPLC (Prep-C18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies; gradient elution of 35% MeCN în water to 60% MeCN in water over a 10 min perîod, where both solvents contain 0.1% FA). This resulted in 3-[3-[(3S)-3-(2-formyl-3-hydroxyphenoxymethyl)morpholine-4carbonyl]pyrazin-2-yl]propanenitrile. LCMS (ES) [M+l]⁺ m/z 397.2. Ή NMR (300 MHz, DMSOAppm) δ 11.75 (s, 1H), 10.32 (s, 1 H), 8.72 (dd,J = 8.2, 2.5 Hz, 1 H), 8.55 (dd,7= 13.7, 15 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).

139

Example 29. 2-hydroxy-6-{[(3S)-4-[3-(hydroxymethyl)pyrazine-2-carbonyl]morpholin-3yl]methoxy}benzaldehyde, Compound 33

[0352] Compound 33 was synthesized according to Scheme 29.

Scheme 29

Step 5

Step 6

Step 7

Step 1

[0353] Into a 250-mL round-bottom flask purged and maîntained with an inert atmosphère of nitrogen, was placed methyl 3-bromopyrazine-2-carboxylate (5.00 g, 23.039 mmol, 1.00 equiv), H2O (100.00 mL). This was followed by the addition of NaBI-L (4.36 g, 115.243 mmol, 5.00 equiv) in several batches at 0 ^rtC. The resulting solution was stirred for overnight at room température. The reaction was quenched by the addition of 50 mL of EtOH and diluted with 150 mL of K₂CO₃(_aq.). The,n the mixture was stirred for 0.5 b at room température. The resulting solution was extracted with 3 x 150 mL of ethyl acetate and extracted with 3 x 150 mL of dichloromethane; the organic layer was dried over anhydrous sodium sulfate and concentrated.

This resulted in (3-bromopyrazin-2-yl)methanol. LCMS (ES) [M+l]⁺ m/z 189.1.

140

Step 2

[0354] Into a 100-mL round-bottom flask, was placed (3-bromopyrazin-2-yl)methanol (3.00 g, 15.872 mmol, 1.00 equiv), DCM (60.00 mL), imidazole (2.16 g, 31.729 mmol, 2.00 equiv), and TBSC1 (2.87 g, 19.042 mmol, 1.20 equiv). The resulting solution was stirred for 3 h at room température and diluted with 50 mL of H₂O. The resulting solution was extracted with 3 x 150 mL of dichloromethane; the organic layer was drîed over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/pelroleum ether (20%EA) as eluents. This resulted in 2-bromo-3-[[(tertbutyldimethylsilyl)oxy]methyl]pyrazine. LCMS (ES) [M+l]⁺ m/z 303.1.

Step 3

[0355] Into a 250-mL pressure tank reactor, was placed 2-bromo-3-[[(tertbutyldimethylsilyl)oxy]methyl]pyrazine (6.00 g, 19.784 mmol, 1.00 equiv), Pd(dppf)Cl₂ (1.45 g, 1.978 mmol, 0.10 equiv), TEA (6.01 g, 59.352 mmol, 3.00 equiv), MeOH (100.00 mL), and CO (gas). The resulting solution was stirred for overnight at 90 ^ÜC. The resulting mixture was concentrated. The residue was purified by silica gel column chromatograhy with PE/THF (70% THF) as eluents. This resulted in methyl 3-[[(tert-butyidimethylsilyl)oxy]methyl]pyrazine-2carboxylate. LCMS (ES) [M+l]⁺ m/z 283.2.

Step 4

[0356] Into a 250-mL round-bottom flask, was placed methyl 3-[[(tert20 butyldimethylsilyl)oxy]methyI]pyrazine-2-carboxylate (3.10 g, 10.977 mmol, 1.00 equiv), melhanol (50.00 mL). This was followed by the addition of a solution of LiOHH₂O (0.92 g, 21.924 mmol, 2.00 equiv) in H₂O (10 mL) dropwise with stirring at 0°C in 5 min. The resulting solution was stirred for 5 h at room température. The pH value of the solution was adjusted to 5 with citric acid (2 mol/L). The resulting solution was extracted with 5 x 150 mL of dichloromethane; the organic layer was dried over anhydrous sodium sulfate and concentrated. This resulted in 3-[[(tert-butyldimethylsilyl)oxy]methyl]pyrazine-2-carboxylic acid. LCMS (ES) [M+l]⁺ m/z 269.2.

Step 5

[0357] Into a 20-mL vial, was placed 3-[[(tert-butyldimethylsiiyI)oxy]methyl]pyrazine-230 carboxylic acid (1.00 g, 3.726 mmol, 1.00 equiv), (3R)-morpholin-3-ylmethanol hydrochloride (0.68 g, 4,471 mmol, 1.20 equiv), dimethylformamide (10.00 mL), HATU (1.70 g, 4.471 mmol,

141

1.20 equiv), and DIEA (1.95 mL, 15.064 mmol, 3.00 equiv). The resulting solution was stirred for 3 h at room température. The resulting solution was diluted with 50 mL of H₂O. The resulting solution was extracted with 4 x 60 mL of dichloromethane, and the organic layer was washed with 2 x 100 mL of brine. The mixture was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography with PE/THF (60%THF) as eluents. This resulted in [(3R)-4-(3-[[(tertbutyldimethylsilyl)oxy]methyl]pyrazine-2-carbonyl)morpholin-3-yl]methanol. LCMS (ES) [M+lf m/z 368.2.

Step 6

[0358] Into a 100-mL 3-necked round-bottom flask purged and maintaîned with an inert atmosphère of nitrogen, was placed [(3R)-4-(3-[[(tert-butyldiraethylsilyl)oxy]methyl]pyrazine-2carbonyI)raorpholin-3-yl]methanol (0.97 g, 2.639 mmol, 1.00 equiv), 2,6dihydroxybenzaldehyde (0.73 g, 5.279 mmol, 2.00 equiv), PPh₃ (1.38 g, 5.261 mmol, 1.99 equiv), and THF (60 mL). The resulting solution was stirred for 15 min at 0 °C. This was followed by the addition of DI AD (1.07 g, 5.279 mmol, 2.00 equiv) dropwise with stirring at 0 °C in 2 min. The resulting solution was stirred for 2 h at room température. The resulting mixture was concentrated. The residue was purified by silica gel column with PE/THF (50%THF) as eluents. This resulted in 2-([(3S)-4-(3-[[(tertbutyldimethylsilyl)oxy]methyl]pyrazine-2-carbonyl)morpholin-3-yl]methoxy]-620 hydroxybenzaldehyde. LCMS (ES) [M+l]⁺ m/z 488.2.

Step 7

[0359] Into a 100-mL round-bottom flask, was placed 2-[[(3S)-4-(3-[[(tertbutyldimethylsilyl)oxy]methyl]pyrazine-2-carbonyl)morpholîn-3-yl]methoxy]-6hydroxybenzaldehyde (0.8 g, 1.641 mmol, 1.00 equiv), THF (20.00 mL), and TBAF (2.5 mL, 25 1.5 equiv, 2M). The resulting solution was stirred for 2 h at room température. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with (60%THF) as eluents. The collected product was further purified by Flash-Prep-HPLC (PrepC18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies; gradient eiution of 30% MeCN in water to 50% MeCN in water over a 10 min period, where both solvents contain 0.1% FA). This resulted in 2-hydroxy-6-{[(3S)-4-[3-(hydroxymethyl)pyrazine-2-carbonyl]morpholin-3yl]methoxy}benzaldehyde. LCMS (ES) [M+l]⁺ m/z 374.1. Ή-NMR (300 MHz, DMSO-<i_ôt ppm) δ 11.79 (s, 1 H), 10.29 (d, J - 3.6 Hz, 1H), 8.64 (t, J = 2.4 Hz, 1H), 8.54 (dd, J = 12.2, 2.6

142

Φ Hz, 1Η), 7.62-7.49 (m, 1H), 6.77-6.48 (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)-l-[2-(l,2-dihydroxyethyl)benzoyl]piperidin-2-yl]niethoxy}-6hydroxybenzaldehyde, Compound 34

[0360] Compound 34 was synthesized according to Scheme 30.

Scheme 30

Step 1

[0361] Into a 100-mL 3-necked round-bottom flask, was placed o-bromobenzoic acid (5.0 g, 10 24.87 mmol, 1.0 equiv), (2S)-2-[[(tert-butyldimethylsilyl)oxy]meihyl]piperidine (6.90 g, 30.07 mmol, 1.2 equiv), DCM (50.0 mL), and DIEA (6.50 g, 50.29 mmol, 2.0 equiv). This was followed by the addition of HATU (11.40 g, 29.98 mmol, 1.2 equiv) ai 0 °C. The reaction solution was stirred for 2 h at room température. The reaction was then quenched by the addition of water (30 mL), and extracted with 2x50 mL of dichloromethane. The residue was purified by 15 silica gel coiumn with ethyl acetate/petroleum ether (10%) as eluents. (S)-(2-bromophenyl)(2(((tert-butyldimethylsilyl)oxy)methyl)piperîdin-l-yl)methanone was obtained. LCMS (ES) [M+l]⁺ m/z: 412.

143

Step 2

[0362] Into a 250-mL round-bottom flask, was placed (S)-(2-bromophenyl)(2-(((tertbutyldimethy!silyl)oxy)methyl)piperidin-l-y!)methanone (5,0 g, 12.12 mmol, 1.0 equiv), THF (50 mL), and 1 M TBAF in THF (12.1 mL, 12.12 mmol, 1.0 equiv). The mixture was stirred for 5 2 h at room température. The mixture was concentrated to remove the solvent, and the residue was purified by silica gel column with ethyl acetate (100%) as eluents. This resulted in (S)-(2biOmophenyl)(2-(hydroxymethyl)pÎperÎdin-l-yl)methanone. LCMS (ES) [M+l]⁺ m/z: 298.

Step 3

[0363] Into a 250-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed (S)-(2-bromophenyl)(2-(hydroxymethyl)piperidin-1 yl)methanone (2.0 g, 6.71 mmol, 1.0 equiv), 2,6-dihydroxybenzaldehyde (1.12 g, 8.11 mmol, 1.2 equiv), THF (80 mL), and PPh₃ (2.10 g, 8.01 mmol, 1.2 equiv). This was followed by the addition of DIAD (1.63 g, 8.05 mmol, 1.2 equiv) at 0 °C. The réaction solution was stirred overnight at room température. The solution was concentrated to remove the solvent, and the 15 residue was purified by silica gel column with ethyl acetate/petroleum ether (80%) as eluents.

This resulted in (S)-2-((L(2-bromobenzoyl)piperidin-2-yl)methoxy)-6-hydroxybenzaldehyde.

LCMS (ES) [M+l]⁺ m/z: 418.

Step 4

[0364] Into a 50-mL round-bottom flask purged and maintained with an inert atmosphère of 20 nitrogen, was placed (8)-2-(( 1-(2-bromobenzoyl)piperidin-2-yl)methoxy)-6hydroxybenzaldehyde (1.57 g, 3.75 mmol, 1.0 equiv), dioxane (20 mL), tributyl(ethenyl)stannane (2.40 g, 7.54 mmol, 2.0 equiv), and Pd(dppf)Ch (307 mg, 0.37 mmol, 0.10 equiv). 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 with ethyl 25 acetate/petroleum ether (1/1) as eluents. This resulted in (S)-2-hydroxy-6-((l -(2vinylbenzoyl)piperidin-2-yl)methoxy)benzaldehyde. LCMS (ES) [M+l]⁺ m/z: 366.

Step 5

[0365] Into a 20-mL vial, was placed (S)-2-hydroxy-6-((l-(2-vinylbenzoyl)piperidin-2yl)methoxy)benzaldehyde (604 mg, 1.65 mmol, 1.0 equiv), r-BuOH (4.0 mL), H₂O (4.0 mL), 30 and ad-mix-alpha (2.60 g, 4.96 mmol, 3.0 equiv). The mixture was stirred for 3 h at room température. The crude product was purified by Prep-HPLC with the following conditions

144 (SH1MADZU (HPLC-01)): Column, Kinetex EVO Cl 8 Column, 21.2*150,5 uni, mobile phase, Water (0.1% FA) and CH3CN (45% Phase B up to 65% in 9 min), Detector, UV 254 nm. 2{[(2S)-l-[2-(l,2-dihydroxyethyl)bcnzoyl]piperidin-2-yl]methoxy}-6-hydroxybenzaldehyde was obtained. LCMS: (ES, m/z); [M+H] ⁺: 400.2. *H-NMR: (300 MHz, DMSO-c/sppm): 611.73 (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-{[(3R)-4-{2-[(lS)-l,2-dihydroxyethyl]benzoyl}thiomorphoIin-3-yl]methoxy}6-hydroxybenzaldehyde and 2-{[(3R)-4-{2-[(lR)-l,2dihydroxyethyl]benzoyI}thiomorpholin-3-yI]methoxy}-6-hydroxybenzaldehyde

[0366] Compound 35, Diastereomer 1 and Compound 35, Diasteroemer 2 were synthesized according to Scheme 31.

Scheme 31

â A AA™ H Step 1 oV 31a ' chirai-HPLC Y Step 3 / O

Λ OH 'NA [Y N | CHO _. YvA 0H_- aA °Y VOH ί A, step 2 LA a a° ov °A 3113 31c A. As γΑΑί Av Av xo 0 Ό + Q 0 9 _r A^xch° Ah0 ' L ï LA Compound 31d Diastereomer 1 and Diastereomer 2 Step 4a or Step 4b V V AO AA A 0 v + , °h° 1 0H 1 CHO OH AvCHO OH Y\^CHO U II \ il kA Y V OH Compound 35 Diastereomer 1 and Diastereomer 2

145

Step 1

[0367] Into a 100-mL round-bottom flask, was placed 2-(2,2-dimethyl-l,3-dioxolan-4yl)benzoic acid (900 mg, 4.05 mmol, 1.00 equiv), DMF (10.0 mL), (3R)-thiomorpholin-3ylmethanol (593 mg, 4.45 mmol, 1.10 equiv), and DIEA (1.05 g, 8.09 mmol, 2.00 equiv). This 5 was followed by the addition of HATU (2.31 g, 6.07 mmol, 1.50 equiv) at 0°C. The resulting solution was stirred for 2 hr at room température. The resulting solution was diluted with 50 mL of H₂O and extracted with 3x30 mL of ethyl acetate, and the organic layers were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1/10) as eluents. This resulted in

[(3R)-4-[2-(2,2-dimethyl-l,3-dioxolan-4-yl)benzoyl]thiomorphoIin-3-yl]methanol. [M+l]⁺ m/z: 338.1.

Step 2

[0368] Into a 50-mL round-bottom flask, was placed [(3R)-4-[2-(2,2-dimethyl-l,3-dioxoIan-4yl)benzoyl]thiomorpholin-3-yl]methanol (620 mg, 1.83 mmol, 1.00 equiv), DCE (8.0 mL), and 15 DIEA (1.42 g, 11.02 mmol, 6.00 equiv). This was followed by the addition of MsCl (420 mg, 3,67 mmol, 2.00 equiv) dropwise with stirring at 0 °C. The resulting solution was stirred for 1 hr at room température. To this solution was added 2,6-dihydroxybenzaldehyde (253 mg, 1.83 mmol, 1.00 equiv). The resulting solution was stirred for 6 hr at 80 °C. The reaction mixture was cooled and concentrated. The resulting solution was diluted with 5 mL of ACN. The crude product was purified by Flash-Prep-HPLC with the following conditions (IntelFlash-1): Column, Cl8 silica gel; mobile phase, H₂O (0.1%HCOOH)/ACN=1/1 încreasing to H₂O(0.1%HCOOH)/ACN=l/2 within 10 min; Detector, UV 254 nm. This resulted in 2-[[(3R)4-[2-(2,2-dimethyl-l,3-dioxolaii-4-yl)benzoyl]thiomorpholm-3-yl]methoxy]-6hydroxybenzaldehyde. [M+l] * m/z: 458.2.

Step 3

[0369] 2-hydroxy-6~[[(3R)-4-[2-(2-methyl-l,3-dioxolan-4-yl)benzoyi]thiomorpholin-3yl]methoxy] benzaldehyde was purified by Chiral-Prep-HPLC with the following conditions: Mobile phase A: n-Hexane; Mobile phase B:EtOH; Flow rate: 20 mL/min; Column: DA1CEL CHIRALPAK ID, 250*20mm, 5um; Gradient;5%B in 20min; Detector, UV 254 nm. The collected products were subjected to analytical chiral HPLC analysis (Instrument Name: Shimadzu LC-20AD; Mobile Phase A: n-Hexane; Mobile Phase B: Ethanol; Column: CHIRALPAK IC-3, 50*4.6mm, 3um IC30CC-SC002).This resulted in Compound 31d,

146

Diastereomer 2 (Analytîcal HPLC Rétention Time = 2.188 min) and Compound 31d, Diastereomer 1 (Analytîcal HPLC Rétention Time = 2.988 min).

Step 4A: Compound 35, Diastereomer 1

[0370] Into a 50-mL round-bottom flask, was placed Compound 31, Diastereomer 1 (80 mg, 5 0.17 mmol, 1.00 equiv), ACN (2.0 mL), Yb(OTf)₃ H₂O (54 mg, 0.08 mmol, 0.50 equiv). The resulting solution was stirred for 5 hr at room température. The resulting solution was diluted with 5 mL of ACN and filtered. The crude product was purified by Prep-HPLC with the foliowîng conditions (2#SHIMADZU (HPLC-01)): Column, Welch Xtimate C18, 21.2*250mm,5um; mobile phase, Water and ACN (15% Phase B up to 70% in 20 min);

Detector, UV 254 nm.. The product was analyzed by chiral SFC (Instrument Name: Shimadzu LC-30AD SF; Column: AS-3,100*3mm). This resulted in Compound 35, Diastereomer 1. SFC rétention time = 2.75 min. LCMS [M+l] ⁺ m/z: 418.2. Ή NMR (300 MHz, DMSO-î/,) δ 11.76 (br, 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), 4.934.37 (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).

Step 4B: Compound 35, Diastereomer 2

[0371] Into a 50-mL round-bottom flask was placed Compound 31d, Diastereomer 2 (90 mg, 0.19 mmol, 1.00 equiv), ACN (2.0 mL), and Yb(OTf)₃H₂O (61 mg, 0.09 mmol, 0.50 equiv). The resulting solution was stirred for 5 hr at room température. The resulting solution was diluted with 5 mL of ACN and filtered. The crude product was purified by Prep-HPLC with the 20 following conditions (2#SHIMADZU (HPLC-01)): Column, Welch Xtimate C18, 2L2*250mm,5um; mobile phase, Water and ACN (15% Phase B up to 70% in 18 min);

Detector, UV 254 nm.. The product was analyzed by chiral SFC (Instrument Name: Shimadzu LC-30AD SF; Column: AS-3, 100*3mm). This resulted in Compound 35, Diastereomer 2. SFC rétention time = 2.44 min. LCMS [M+l]⁺ m/z: 418.2. ^]H NMR (300 MHz, DMSO-î/ô) δ 11.76 25 (br, 1H), 10.46-10.03 (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).

147

Example 32. 2-{[(2S)-l-[2-(l,2-dihydroxyethyl)pyridine-3-carbonyl]piperidin-2yi]methoxy}-6-hydroxybenzaldehyde, Compound 36

[0372] Compound 36 was synthesized according to Scheme 32.

Scheme 32

32d

Step 1

[0373] Into a 100-mL 3-necked round-bottom flask, was placed 2-bromopyridine-3-carboxylîc acid (4.0 g, 19.80 mmol, 1.0 equiv), (2S)-2-[[(tert-butyIdimethylsilyl)oxy]methyl]piperidine (5.50 g, 23.97 mmol, 1.2 equiv), DCM (50 mL), and DIEA (5.13 g, 39.70 mmol, 2.0 equiv).

This was followed by the addition of HATU (9.07 g, 23.85 mmol, 1.2 equiv) at 0 ^lC. The reaction solution was stirred for 2 h at room température. The reaction was then quenched by the addition of water (50 mL), and extracted with 2x50 mL of dichloromethane. The combined organic phase was dried over anhydrous sodium sulfate and filtered, and the fîltrate was concentrated under reduced pressure. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/4). (S)-(2-bromopyrîdin-3-yl)(2-(((tertbutyldimethylsilyl)oxy)methyl)piperidin-l-yi)methanone was obtained. LCMS (ES) [M+l]⁺ m/z: 413.

Step 2

[0374] Into a 250-mL round-bottom flask, was placed (S)-(2-biOmopyridin-3-yl)(2-(((tert20 bulyldimethylsily!)oxy)methyl)piperidin-l-yl)melhanone (8.0 g, 19.35 mmol, 1.0 equiv), THF

1.48 (80 mL), and TB AF (1 M in THF) (20 mL, 20.0 mmol, 1.0 eq). The mixture was stirred for 2 h at room température and concentrated to remove the solvent. The residue was purified by silica gel column chromatography with ethyl acetate (100%) as eluents. This resulted in (S)-(2bromopyridin-3-yl)(2-(hydroxymethyl)piperidin-l-yl)methanone. LCMS (ES) [M+l]⁺ m/z: 299.

Step 3

[0375] Into a 250-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed (S)-(2-bromopyridin-3-yl)(2-(hydroxymethyl)piperidin-lyl)methanone (2.0 g, 6.69 mmol, 1.0 equiv), 2,6-dihydroxybenzaldehyde (1.10 g, 7.96 mmol, 1.2 equiv), PPh₃ (2.10 g, 8.01 mmol, 1.2 equiv), and THF (80 mL). This was followed by the 10 addition of DIAD (1.63 g, 8.06 mmol, 1.2 equiv) at 0 °C. After addition, the resulting solution was stirred overnîght al room température. The mixture was concentrated to remove the solvent, and the residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1/1) as eluents. This resulted in 2-(S)-2-((l-(2-bromonicotinoyl)piperidin-2-yl)methoxy)6-hydroxybenzaldehyde. LCMS (ES) [M+l]⁺ m/z: 419.

Step 4

[0376] Into a 100-mL round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed 2-(S)-2-((i-(2-bromonicotinoyl)piperidin-2-yl)methoxy)-6hydroxybenzaldehyde (3.20 g, 7.63 mmol, 1.0 equiv), dioxane (30 mL), tributyl(ethenyl)stannane (4.85 g, 15.30 mmol, 2.0 equiv), and Pd(dppf)Ch'CH2C12 (624 mg, 20 0.76 mmol, 0.10 equiv). The mixture was stirred overnîght at 90 °C. After cooiing to room température, the reaction solution was concentrated to remove the solvent, and the residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1/1) as eluents. This resulted in (S)-2-hydroxy-6-((l -(2-vinylnicotinoyl)piperidin-2-yl)methoxy)benzaidehyde.

LCMS (ES) [M+l]⁺ m/z: 367.

Step 5

[0377] Into a 100-mL round-bottom flask, was placed (S)-2-hydroxy-6-(( 1 -(2vinylnicotinoyl)piperidin-2-yl)methoxy)benzaldehyde (500 mg, 1.37 mmol, 1.0 equiv), t-BuOH (20.0 mL), H2O (20.0 mL), and AD-mix-alpha (5.31 g, 6.82 mmol, 5.0 equiv). The mixture was stirred overnîght at room température. The mixture was concentrated to remove the solvent, and 30 the crude product was purified by Prep-HPLC wîth the following conditions (SHIMADZU (HPLC-01): Column, Kînelex EVO C18 Column, 21.2*150, 5 um, mobile phase, Water (0.1%

149

FA) and CH3CN (45% Phase B up to 65% in 9 min), Detector, UV 254 nm. This resulted in 2{[(2S)-l-[2-(l,2-dihydroxyethyI)pyridine-3-carbonyi]piperidin-2-yl]methoxy}-6hydroxybenzaldehyde. LCMS: (ES, m/z): [M+H]⁺: 401.2. 'H-NMR (300 MHz, DMSO-r/sppnz): 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), 6.785 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), 2.081.50(m, 6H).

Example 33. 2-hy(lroxy-6-{[(3R)-4-[2-(2-hydroxyethyl)pyndine-3-carbonyl]morpholin-3yl]methoxy}benzaldehyde, Compound 37

[0378] Compound 37 was synthesized according to Scheme 33.

33c

Step 1

[0379] Into a 50-mL 3-necked round-bottom flask, was placed 2-[2-[(tertbutyldimethylsilyl)oxy]ethyl]pyridine-3-carboxylic acid (1.50 g, 5.33 mmol, 1.0 equiv), (3S)15 morpholin-3-ylmethanol hydrochloride (980 mg, 6.38 mmol, 1.2 equiv), DCM (15 mL), and

DIEA (2.07 g, 16.02 mmol, 3.0 equiv). HATU (2.40 g, 6.31 mmol, 1.2 equiv) was added by 3 batches at 0 C. After addition, the mixture was stirred for 2 h at room température. The reaction was then quenched by the addition of water (20 mL), and extracted with 3x20 mL of dîchloromethane. The combined organic phase was dried over anhydrous sodium sulfate and

150 filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (80%) as eluents. This resulted in (S)-(2-(2-((tert-butyldimelhylsilyl)oxy)ethyl)pyridin-3-yl)(3(hydroxymethyl)morpholino)methanone. LCMS (ES) [M+l]⁺ m/z: 381.

Step 2

[0380] Into a 250-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed (S)-(2-(2-((tert-butyldimethylsilyl)oxy)ethyl)pyridin-3-yl)(3(hydroxymethyl)morpholino)methanone (1.98 g, 5.20 mmol, 1.0 equiv), 2,6- dihydroxybenzaldehyde (863 mg, 6.25 mmol, 1.2 equiv), PPha (1.64 g, 6.25 mmol, 1.2 equiv), 10 and THF (80 mL). After coolîng to 0°C, DBAD (1.44 g, 6.25 mmol, 1.2 equiv) was added in one portion. The mixture was stirred overnight at room température. The mixture was concentrated to remove the solvent, and the residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (80%) as eluents. This resulted in (R)-2-((4-(2-(2-((tertbutyldimethyisilyi)oxy)ethyI)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybenzaIdehyde.

LCMS (ES) [M+l]⁺ m/z: 501.

Step 3

[0381] Into a 20-mL vial, was placed (R)-2-((4-(2-(2-((tertbutyldimethylsilyl)oxy)ethyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybenzaldehyde (300 mg, 0.60 mmol, 1.0 equiv), CH₃CN (5.0 mL), and HCOOH (1.0 mL). The reaction solution was 20 stirred for 1 h at 50 °C and then concentrated to remove the solvent. The residue was purified by

Prep-HPLC with the following conditions: Kînetex EVO C18 column, 21.2*150, 5 um, mobile phase, Water (0.1% FA) and CH₃CN (10% Phase B up to 50% within 15 min), detector, UV 254 nm. This resulted in 2-hydroxy-6-{[(3R)-4-[2-(2-hydroxyethyl)pyridine-3-carbonyl]morphoiin3-yl]methoxy}benzaldehyde. LCMS (ES, mlz): [M+H]⁺: 387.1. 'H-NMR (300 MHz, DMSO-î/ô, 25 ppm): δ 11.74 (br, IH), 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), 5.03-4.89 (m, 1H), 4.65- 4.34 (m, 3H), 4.11-3.34 (m, 7H), 3.11-2.84 (m, 3H).

151

Example 34. 2-hydroxy-6-Î[(2R)-l-[2-(2-hydroxyethyl)pyridine-3-carbonyl]piperidin-2yl]methoxy}benzaldehyde, Compound 38

[0382] Compound 38 was synthesized according to Scheme 34.

Scheme 34

Step 1

[0383] Into a 50-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed 2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridine-3carboxylic acid (1.00 g, 3.55 mmol, 1.00 equiv), DMF (25.0 mL), (2R)-piperidin-2-ylmethanol (491 mg, 4.26 mmol, 1.20 equiv), and DIEA (551 mg, 4.26 mmol, 1.20 equiv). This was followed by the addition of HATU (1.62 g, 4.26 mmol, 1.2 equiv) in several batches at 0 °C. The reaction solution was stirred overnight at room température. The reaction was diluted with 30 mL of H₂O and extracted with 3 x 100 mL of ethyl acetate. The combined organîc phase was washed with 1 x 50 mL of 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 with ethyl acetate/petroleum ether (1:2) as eluents. This resulted in [(2R)-1-(2-(2-[(tert-buty Idimethylsily l)oxy]ethyl]py ridine-3-carbonyl)piperidin-2yl]methanol. LCMS (ES) [M+l]⁺ m/z: 379.

152

Step 2

[0384] Into a 100-inL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed [(2R)-l-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridine-3carbonyl)piperidin-2-yl]methanol (1.20 g, 3.17 mmol, 1.00 equiv), THF (50.0 mL), 2,6- dihydroxybenzaldehyde (525 mg, 3.80 mmol, 1.20 equiv), and PPh₃ (998 mg, 3.80 mmol, 1.20 equiv). This was followed by the addition of a solution of DIAD (769 mg, 3.80 mmol, 1.20 equiv) in THF (2.00 mL) dropwise with stirring at 0 ^ÜC. The reaction solution was stirred overnight at room température. After concentrating under reduced pressure, the residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1:1) as eluents.

This resulted in 2-[[(2R)-l-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridme-3carbonyl)piperidin-2-yl]methoxy]-6-hydroxybenzaldehyde. LCMS (ES) [M+l]⁺ m/z: 499.

Step 3

[0385] Into a 50-mL round-bottom flask, was placed 2-[[(2R)-l-(2-[2-[(tertbutyldimethylsilyl)oxy]ethyl]pyrîdÎne-3-carbonyl)piperidin-2-yl]methoxy]-615 hydroxybenzaldehyde (300 mg, 0.60 mmol, 1.00 equiv), CH3CN (5.00 mL), and HCOOH (1.00 mL). The mixture was stirred for 3 h at 50 °C in oil bath. The reaction mixture was cooled to room température and concentrated under reduced pressure. The crude product was purified by Flash-Prep-HPLC with the following conditions (IntelFlash-1): Column: Ascentis Express C18, 50*3.0 mm, 2.7 um, Mobile Phase A: Water/0.05% FA, Mobile Phase B: CH₃CN; Flow rate: 1.5 mL/min, Gradient: 5%B to 100%B in 1.2 min, hold 0.6 min. This resulted in 2-hydroxy-6{[(2R)-l-[2-(2-hydroxyethyl)pyridine-3-carbonyl]pîperidin-2-yl]methoxy}benzaldehyde. LCMS (ES, m/z): [M+H] ⁺: 385. ‘H-NMR (300 MHz, DMSO-i^ppm): δ 11.73 (br, 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), 1.95-1.39 (m, 6H).

153

Example 35. 2-hydroxy-6-{[(3S)-4-[2-(2-hydroxy-2-methylpropyl)pyridine-3 carbonyl]morphoIm-3-yl]methoxy}benzaldeliyde, Compound 39

[0386] Compound 39 was synthesized according to Scheme 35.

Scheme 35

Step 1

[0387] Into a 1-L 3-necked round-bottom flask purgcd and maintained with an inert atmosphère of nitrogen, was placed 3-bromo-2-methylpyridîne (20.0 g, 116.26 mmol, 1.0 equiv) and THF (400 mL). This was followed by the addition of LDA (2M in THF) (69.8 mL, 139.51 mmol, 1.2 equiv) at -78 °C and stirred for 0.5 h. To this was added acetone (7.46 g, 128.45 mmol, 1.1 equiv) at the same température. The mixture was stirred for 1 hr at -78 °C. The reaction was then quenched by the addition of NHiClfaq) (300 mL) and extracted with 3x500 mL of ethyl acetate. The combined organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by

154

F silica gel column chromatography with ethyl acetate/petroleum ether (15%) as eluents. This resulted in l-(3-bromopyridin-2-yl)-2-methylpropan-2-ol. LCMS (ES) [M+l]⁺ m/z: 230.

Step 2

[0388] Into a 250-mL round-bottom flask, was placed l-(3-bromopyridin-2-yl)-25 methylpropan-2-ol (4.0 g, 17.38 mmol, 1.0 equiv), TBSC1 (3.10 g, 20.86 mmol, 1.2 eq), DMF (40 mL), imidazole (2.38 g, 34.76 mmol, 2.0 eq), and DMAP (212 mg, 1.74 mmol, 0.10 equiv). The reaction solution was stirred 24 h at 60 °C. After cooling to room température, the réaction was then quenched by the addition of water (50 mL) and extracted with 3x50 mL of ethyl acetate. The combined organîc phase was washed with 2x50 mL of 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 with ethyl acetate/petroleum ether (1/20) as eluents. This resulted in 3-bromo-2-(2-((tertbutyldimethylsiIyl)oxy)-2-methylpropyl)pyridine. LCMS (ES) [M+l]⁺ m/z: 344.

Step 3

[0389] Into a 250-mL pressure tank reactor, was placed 3-bromo-2-(2-((tertbutyldimethylsilyi)oxy)-2-methylpropyl)pyridine (4.18 g, 12.14 mmol, 1.0 equiv), MeOH (80 mL), TEA (2.45 g, 24.28 mmol, 2.0 equiv), and Pd(dppf)Ch CH2CI2 (495 mg, 0.61 mmol, 0.05 eq). The mixture was stirred for 12 h at 130 °C under CO(_g) atmosphère at 30 atm. The mixture was concentrated to remove the solvent; the residue was purified by silica gel column chromatography with ethyl acetate/petroleum (1/3) as eluents. This resulted in methyl 2-(2((tert-butyldimethylsilyl)oxy)-2-methylpropyl)mcotinate. LCMS (ES) [M+1J+ m/z: 324.

Step 4

[0390] Into a 50-mL round-bottom flask, was placed methyl 2-(2-((tertbutyldimethylsilyI)oxy)-2-methylpropyl)nicotinate (2.0 g, 6.18 mmol, 1.0 equiv), MeOH (16 25 mL), and H₂O (8 mL). This was followed by the addition of LiOH (520 mg, 12.26 mmol, 2.0 equiv) at 0 °C. The mixture was stirred for 2 h at 50 °C. After cooling to room température, the pH value of the solution was adjusted to 7 with ci trie acid. The solids were collected by filtration and dried under infrared lamp. This resulted in 2-(2-((tert-butyldimethyIsilyl)oxy)-2methylpropyl)nicotinic acid. LCMS (ES) [M+l]⁺ m/z: 310.

155

Step 5

[0391] Into a 50-mL 3-necked round-bottom flask, was placed 2-(2-((tertbutyldimethylsilyl)oxy)-2-methylpropyl)nicotinic acid (1.60 g, 5.17 mmol, 1.0 equiv), (3R)morpholin-3-ylmethanol hydrochloride (951 mg, 6.19 mmol, 1.2 equiv), DMF (16 mL), and

DIEA (2.0 g, 15.48 mmol, 3.0 equiv). This was followed by the addition of HATU (2.36 g, 6.21 mmol, 1.20 equiv) at 0 °C. The reaction solution was stirred for 2 h at room température. The reaction was then quenched by the addition of water (30 mL) and extracted with 3x30 mL of ethyl acetate. The combined organic phase was washed with brîne (30 mLx3) and dried over anhydrous sodium sulfate. The mixture was filtered, and the fîltrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1/1) as eluents. This resulted in (R)-(2-(2-((tertbutyldimethylsilyl)oxy)-2-methyIpropyl)pyridin-3-yI)(3(hydroxymethyl)morphoiino)methanone. LCMS (ES) [M+l]⁺m/z; 409.

Step 6

[0392] Into a 100-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed (R)-(2-(2-((tert-butyldimethylsilyl)oxy)-2methylpropy!)pyridin-3-yi)(3-(hydroxymethyl)morpholino)methanone (1.0 g, 2.45 mmol, 1.0 equiv), 2,6-dihydroxybenzaldehyde (406 mg, 2.94 mmol, 1.2 equiv), PPfo (770 mg, 2.94 mmol, 1.2 equiv), and THF (50 mL). This was followed by the addition of DIAD (594 mg, 2.94 mmol, 20 1.2 equiv) at 0 °C. The mixture was stirred overnight at room température. The mixture was concentrated to remove the solvent, and the residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1/1). This resulted in (S)-2-((4-(2-(2-((tertbutyldimethylsilyl)oxy)-2-methylpropyl)nicotinoyl)morpholin-3-yl)methoxy)-6hydroxybenzaldehyde. LCMS (ES) [M+l]⁺ m/z: 529.

Step 7

[0393] Into a 20-mL vial, was placed (S)-2-((4-(2-(2-((tert-butyldimethylsilyl)oxy)-2methylpropyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybenzaldehyde (600 mg, 1.14 mmol, l.Ü equiv), CH₃CN (5.0 mL), and HCOOH (1.0 mL). The mixture was stirred for 1 h at 50 °C. After cooling to room température, the reaction solution was dîrectly purified by Prep30 HPLC with conditions: Column, Ascentis Express C18,50*3.0 mm, 2.7 um, Mobile Phase A:

Water/0.05% FA, Mobile Phase B: CH3CN, Flow rate: 1.5 mL/min, Gradient: 5%B to 100% B in 1.2 min, hold 0.6 min. This resulted in (S)-2-hydroxy-6-((4-(2-(2-hydroxy-220663

156 methylpropyI)nicotinoyl)morpholin-3-yl)methoxy)benzaldehyde. LCMS (ES, m/z): [M+H]'¹·;

415.2. ‘H-NMR(300 MHz, DMSO-do,ppm): δ 11.75 (br, 1H), 10.20 (s, lH),8.57(s, 1H),7.787.28 (ni, 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), 3.152.59 (m, 3H), 1.31-0.92 (m, 6H).

Example 36. 2-hydroxy-6-({4-[2-(hydroxymethyl)benzoyl]thÎomorpholin-3yl}methoxy) benzaldehyde, Compound 40

[0394] Compound 40 was synthesized accordîng to Scheme 36.

Scheme 36

Step 1

[0395] Into a 500-mL 3-necked round-bottom flask, was placed 2-hydroxymethylbenzoic acid (10.0 g, 65.7 mmol, 1.00 equiv), îmîdazole (8.95 g, 131 mmol, 2.00 equiv), and DCM (200 mL). To this solution was added TBDPS-C1 (21.6 g, 78.8 mmol, 1.20 equiv) dropwise at 0 °C. The resulting solution was stirred for 16 hr at 0-25 °C. The reaction was then quenched by the addition of 50 mL of water. The resulting solution was extracted with 3x100 mL of DCM. The organic layer was dried and concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1:50 to 1:1) as eluents. This resulted in 2[[(tert-butyldiphenylsilyl)oxy]methyl]benzoic acid. LCMS (ES) [M+l]⁺ m/z: 391.2.

157

Step 2

[0396] To a solution of 2-[[(tert-butyIdiphenyIsilyl)oxy]methyl]benzoic acid (2.00 g, 5.12 mmol, 1.00 equiv) in DCM (20.0 mL) was added (COC1)₂ (1.30 g, 10.2 mmol, 2.00 equiv) dropwise at 0^aC. The resulting solution was heated to 40 ^l’C for 5 h. The réaction was then concentrated to gîve a residue. The residue was dissolved in THF (20.0 mL), and TEA (1.55 g, 15.3 mmol, 3.00 equiv) was added. To this solution was added thiomorpholin-3-ylmethanol (0.68 g, 5.12 mmol, 1.00 equiv) in portions at 0 °C. The resulting solution was stirred for 16 hr at 0-25 °C. The reaction was then quenched by the addition of 10 mL of water. The resulting solution was extracted with 3x20 mL of ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1:50 to 1:5) as eluents. This resulted in [4-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)thiomorpholin-3-yl]methanol. LCMS (ES) [M+l] ⁺ m/z: 506.7.

Step 3

[0397] Into a 100-mL 3-necked round-bottom flask under N₂ atmosphère, was placed [4-(2[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl)thiomorpholin-3-yl]methanol (1.80 g, 3.55 mmol, 1.00 equiv), 2,6-dihydroxybenzaldehyde (0.74 g, 5.33 mmol, 1.50 equiv), PPh₃ (1.40 g, 5.33 mmol, 1.50 equiv), and DCM (30.0 mL). To this solution was added a solution of DBAD (1.23 g, 5.33 mmol, 1.5.0 equiv) in DCM (3.0 mL) drop wise at 0°C. The resulting solution was stirred for 15 hr at 0-25 °C. The resulting mixture was concentrated. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:100 to 1:1). This resulted in 2-[[4(2-[[(tert-buty 1 diphenylsilyl)oxy]methy 1]benzoyl)thiomorpholin-3-y 1]methoxy]-6-hydro xybenzaldehyde. LCMS (ES) [M+l]⁺ m/z: 626.2

Step 4

[0398] Into a 100-mL 3-necked round-bottom flask, was placed 2-{[4-(2-[[(tertbuty Idiphenylsil y 1) oxy]methyI]benzoyl)thiomorpholîn-3-yl]methoxy]-6-hydroxybenzaldehyde (1.05 g, 1.67 mmol, 1.00 equiv), THF (5.00 mL) and TBAF (0.33 mL, 0.330 mmol, 0.20 equiv). The resulting solution was stirred for 2 hr at 0-25°C. The resulting mixture was concentrated. The residue was purified by silica gel column with ethyl acetate/petroleum ether (50:1-1:1) as eluents. This resulted in 2-hydroxy-6-({4-[2-(hydroxymethyl)benzoyl]thiomorpholin-3yl}methoxy)benzaldehyde. LCMS (ES) [M+Na]⁺ m/z;410.1; Ή NMR (300 MHz, DMSO-ί/ή) S

158

Φ 12.00 (s, 1Η), 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. 2-hy droxy-6-{[(3S)-4-[2-(hydroxy methyl)benzoyl] thiomorpholin-3yl]methoxy}benzaldehyde and 2-hydroxy-6-{[(3R)-4-[25 (hydroxy methy l)benzoy 1] thiomorpholi n-3-y I] methoxy }benzaldehyde

Scheme 37

OH OH OH

Compound 40

Enantiomer 1 and Enantiomer 2

[0399] Compound 40 was purified by Chiral-Prep-HPLC (Conditions: Column: Lux Cellulose4, 4.6*100 mm, 3 pm; mobile phase, A: n-Hexane B: Ethanol (35% B in 18 min); Flow rate: 30 mL/min; Detector, 220 nm) and was analyzed by analytical chiral HPLC (Conditions: instrument name: Shimadzu LC-20AD; Mobile Phase A: n-Hexane/DCM=5/l; Mobile Phase B. Ethanol; Column : CHIRALPAKIA-3, 50*4.6mm, 3um IA30CC-UL005). This resulted in Enantiomer 1 and Enantiomer 2 of Compound 40.

[0400] Compound 40, Enantiomer 1: Analytical chiral HPLC rétention time = 2.42 min;

LCMS (ES) [M+Na]⁺ m/z:410.1; Ή NMR (300 MHz, DMSO-dô) 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), 5.424.41 (m, 6H), 3.47-3.32 (m, 1H), 3.32-2.90 (m, 2H), 2.63-2.51 (m, 2H), 2.50-2.40 (m, 1H).

[0401] Compound 40, Enantiomer 2: Analytical chiral HPLC rétention time = 4.50 min. LCMS (ES) [M+Na]⁺ m/z:410.1; Ή NMR (300 MHz, CDCh) δ 11.98-11.87 (m, 1H), 10.36 (br, 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).

159

Example 38. 2-hydroxy-6-{[(3R)-4-[2-(hydroxymethyl)benzoyl]thioniorphoIin-3y 1] methoxy }benzaldehy de

Scheme 38

Step 1

[0402J Into a 1-L round-bottom flask, was placed phthalide (25.0 g, 186.3 mmol, 1.0 equiv), H₂O (250 mL) and NaOH (14.91 g, 372.762 mmol, 2 equiv). The resulting solution was stirred for 3 h at 100 °C in an oil bath. The reaction mixture was cooled to 0 °C. Solids were precipitated oui after the pH value of the solution was adjusted to 1 with HCl (12mol/L). The 10 solids product was collected by filtrate. This resulted in 2-hydroxymethylbenzoic acid. LCMS (ES) [M+l]⁺ m/z: 153.1. *H NMR (300 MHz, DMSO-rf_fl) 5 12.88 (br, 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).

160

Step 2

[0403] Into a 50Ü-mL 3-necked round-bottom flask, was placed 2-hydroxymethyIbenzoic acid (15.0 g, 98.6 mmol, 1.0 equiv), DCM (200 mL), and imidazole (10.0 g, 147.8 mmol, 1.5 equiv). After the reaction was cooled to 0 °C, TBDPSC1 (32.5 g, 118.3 mmol, 1.2 equiv) was added dropwise with stirring at 0 °C. The resulting solution was stirred for 16 h at 25 °C. The reaction was then quenched by the addition of 300 mL of water. The resulting solution was extracted with 3 x 250 mL of ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1:50 to 1:1) as eluents. This resulted in 2~[[(tert-butyldiphenylsilyl)oxy]methyl]benzoic acid.

LCMS (ES) [M^ 1]⁺ m/z: 391.1. Ή NMR (300 MHz, DMSO-d₆) 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).

Step 3

[0404] Into a 500-mL 3-necked round-bottom flask, was placed 2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoic acid (15.0 g, 38.4 mmol, 1.0 equiv), DCM (250 mL) and 15 two drops of DMF. After the réaction was cooled to 0 °C, (COC1)2 (5.8 g, 46.1 mmol, 1.2 equiv) was added dropwise with stirring at 0 °C. The resulting solution was stirred for 5 h at 40 °C. The mixture was concentrated under vacuum. This resulted in 2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl chloride, which was used for next step without further purification.

Step 4

[0405] To a solution of (3R)-thiomorpholin-3-ylmethanol (5.3 g, 40.3 mmol, 1.05 equiv) and TEA (7.8 g, 76.7 mmol, 2.0 equiv) in THF (250 mL) was added 2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl chloride (15.7 g, 38.3 mmol, 1.0 equiv) in THF (50 mL) dropwise at 0°C. After the addition, the resulting solution was stirred for 5 h at 0-25 °C. The 25 reaction was then quenched by the addition of 50 mL of water. The resulting solution was extracted with 3x150 mL of ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1:50 to 1:5) as eluents. This resulted in [(3R)-4-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl) thiomorpholin-3-yl]methanoL LCMS (ES) [M+l] ⁺ m/z;

506; Ή NMR (300 MHz, DMSO-dr,) 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).

161 * Step 5

[0406] Into a 2.5-L 3-necked round-bottom flask purged and maintained with an inert atmosphère of argon, was placed [(3R)-4-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)thiomorpholin-3-yl]methanol (18.0 g, 35.5 mmol, 5 l.OOequiv), 2,6-dihydroxybenzaldehyde (5.4 g, 39.1 mmol, 1.1 equiv), DCM (900.00 mL) and PPh₃ (14.0 g, 53.3 mmol, 1.5 equiv). This was followed by the addition of DBAD (9.8 g, 42.7 mmol, 1.2 equiv) in DCM (100 mL) dropwise with stirring at 0°C. The resulting solution was stirred for 15 hr at 0-25 °C. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1:100 to 1:1) as eluents.

This resulted in 2-[[(3R)-4-(2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl)thiomorpholin-3yl]methoxy]-6-hydroxybenzaldehyde. LCMS (ES) [M+l] ⁺ m/z: 626.2; Ή NMR (300 MHz, DMSO-Λ) Ô 11.71 (s, 1H), 10.19 (s, 1H), 7.61-7.41 (m, 16H), 6.55 (d, J =8.4 Hz, 1H), 5.235.19 (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).

Step 6

[0407] Into a 5 00-mL 3 -necked round-bottom flask, was placed 2- [[(3R)-4-(2-[ [(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)thîomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde (14.0 g, 22.3 mmol, 1.0 equiv) and THF (140 mL). To this solution was added a solution of TBAF (4.5 mL, 4.50 mmol, 0.20 equiv, 1 M in THF) dropwise with stirring at 0°C. The 20 resulting solution was stirred for 3 hr at 0-25 °C. The resulting mixture was concentrated. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:50 to 1:1) to give crude product. The crude product was purified by Flash-Prep-HPLC with the following conditions (InlelFlash-1): Column, C18 silica gel; mobile phase, MeCN/ H₂O=1:9 increasing to MeCN/ H₂O=1:1 within 15; Detector, 220. This resulted in 2-hydroxy-6-{[(3R)-4-[225 (hydroxymethyl)benzoyl]thiomorpholin-3-yl]methoxy}benzaldehyde, which was subjected to chiral analytical HPLC analysis with the following conditions: Instrument Name: Shimadzu LC20AD; Mobile Phase A: n-Hexane/DCM=5/l; Mobile Phase B: Ethanol; Column: CHIRALPAK IA-3, 50*4.6mm, 3um IA30CC-UL005. Analytical Chiral HPLC rétention time: 4.540 min. LCMS (ES, m/z): [M+Na] ⁺: 410.1; Ή NMR (300 MHz, DMSO-rf₆) 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, 1H).

162

[0408] Based on the product of Scheme 38, it was determined that Compound 40, Enantiomer 2 corresponds to 2-hydiOxy-6-{[(3R)-4-[2-(hydroxymethyl)benzoyl]thiomorpholm-3yl] methoxy }benzal de hy de.

Example 39. 2-{[(2S)-l-{2-[(lR)-l,2-dÎhydroxyethyl]benzoyl}piperidin-2-yl]methoxy}-65 hydroxybenzaldehyde and 2-{[(2S)-l-Î2-[(lS)-l,2-dihydroxyethyl]benzoyl}piperidin-2yl]methoxy}-6-hydroxybenzaldehyde

Scheme 39

Compound 39h

Diastereomer 1 and Diastereomer 2

Step 9a or Step 9b t

Compound 34

Diastereomer 1 and Diastereomer 2

163

Step 1

[0409] Into a 500-mL 3-necked round-bottom flask, was placed 2-bromobenzaldehyde (10.0 g, 54.05 mmol, 1.0 equiv), methyltriphenyl-lambda5-phosphane hydrobromide (23.20 g, 64.58 mmol, 1.2 equiv), and DMF (100 mL). This was followed by the addition of NaH (60% in minerai oil) (9.67 g, 241.69 mmol, 4.5 equiv) carefully at 0 ^ÛC by four batches. The mixture was stirred overnight at room température. The reaction was then quenched by the addition of water/ice (100 mL), extracted with 3x100 mL of 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 with PE (100%) as eluents. This resulted in l-bromo-2vînylbenzene. GCMS:182.

Step 2

[0410] Into a 250-mL round-bottom flask, was placed 1 -bromo-2-vinylbenzene (8.50 g, 46.43 mmol, 1.0 equiv), acetone (130 mL), H₂O (13 mL), NMO (5.43 g, 46.35 mmol, 1.0 equiv), and

K₂OsO4‘2H₂O (730 mg, 2.32 mmol, 0.05 equiv). The mixture was stirred overnight at room température. The mixture was concentrated to remove the solvent, and the residue was purified by silica gel column chromatography with THF/PE (15%) as eluents. This resulted in -(2bromophenyl)ethane-l,2-diol. LCMS (ES) [M+l]⁺ m/z: 217.

Step 3

[0411] Into a 250-mL round-bottom flask, was placed l-(2-bromophenyl)ethane-l,2-diol (5.10 g, 23.50 mmol, 1.0 equiv), 2,2-dimethoxypropane (4.17 g, 40.04 mmol, 1.7 equiv), TsOH (812 mg, 4.72 mmol, 0.20 equiv), and DMF (75 mL). The reaction solution was stirred for 5 h at room température. The reaction was then quenched by the addition of water (100 mL) and extracted with 3x100 mL of 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 with PE (100%) as eluents. This resulted in 4-(2-bromophenyl)-2,2-dimethyl1,3-dioxolane. GCMS: 256.

Step 4

[0412] Into a 100-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed 4-(2-bromophenyl)-2,2-dimethyl-l,3-dioxolane (2.50 g, 9.72

164 mmol, 1.0 equiv), THF (50 mL). This was followed by the addition of n-BuLi (2.5 M in THF) (4.68 mL, 11.68 mmol, 1.2 equiv) at -78 °C. The reaction solution was stirred for 30 min at 78 °C. To this ethyl chloroformate (2.11 g, 19.44 mmol, 2.0 equiv) was added at -78°C. The resulting solution was stirred for additional 1 h at room température. The reaction was then quenched by the addition of NH4CI (aq) (60 mL) and extracted with 3x50 mL of ethyl acetate. The combined organic phase was dried over anhydrous sodium sulfate and fil te red, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether(l:10) as eluents. This resulted in ethyl 2(2,2-dimethyl-l,3-dioxolan-4-yl)benzoate. GCMS: 250.

Step 5

[0413] Into a 100-mL round-bottom flask, was placed ethyl 2-(2,2-dimethyl-l,3-dioxolan-4yl)benzoate (1.60 g, 6.39 mmol, 1.0 equiv), EtOH (10.0 mL), H2O (50.0 mL), and LiOH H₂O (538 mg, 12.82 mmol, 2.0 equiv). The reaction solution was stirred overnight at room température. The mixture was concentrated to remove the solvent, and the pH value of the residue was adjusted to 4 with 2N HCl. The solid was coliected by filtration and dried under infrared lamp. This resulted in 2-(2,2-dimethyl-l,3-dioxolan-4-yl)benzoic acid. LCMS (ES) [ΜΙ]·*· m/z: 221.

Step 6

[0414] Into a 50-mL 3-necked round-bottom flask, was placed 2-(2,2-dimethyl-l,3-dioxolan-420 yi)benzoic acid (865 mg, 3.89 mmol, 1.0 equiv), (2S)-piperîdin-2-ylmethanol (537 mg, 4.66 mmol, 1.2 equiv), DMF (20 mL), and DIEA (1.0 g, 7.74 mmol, 2.0 equiv). This was followed by the addition of HATU (1.78 g, 4.68 mmol, 1.2 equiv) at 0 °C. The mixture was stirred for 2 h at room température. The reaction was then quenched by the addition of water (30 mL) and extracted with 3x20 mL of ethyl acetate. The combined organic phase was dried over anhydrous 25 sodium sulfate and filtered, and the filtrate was concentrated in vacuum. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1/3) as eluents. This resulted in (2-(2,2-dimethy 1-1,3-dioxolan-4-yl)phenyl)((S)-2(hydroxymethyl)piperidîn-l-yl)methanone. LCMS (ES) [M+l]⁺ m/z;320.

Step 7 and Step 8

[0415] Into a 250-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed (2-(2,2-dimethyl-l,3-dioxolan-4-yl)phenyl)((S)-220663

165 (hydroxymethyl)piperidin-l-yl)methanone (900 mg, 2.82 mmol, 1.0 equiv), 2,6dihydroxybenzaldehyde (467 mg, 3.38 mmol, 1.2 equiv), PPh3 (887 mg, 3.38 mmol, 1.20 equiv), and THF (60 mL). This was foliowed by the addition of DIAD (684 mg, 3.38 mmol, 1.2 equiv) at 0 ^ÜC. After addition, the reaction solution was stirred overnight at room température. The mixture was concentrated to remove the solvent, and the residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1/1) as eluents.

[0416] The collected racemate product was separated by Chîral-Prep-HPLC with the following conditions: Mobile phase: A: π-Hexane, B: Ethanol, Flow rate: 20mL/mîn, Column: CHIRALPAK IC-3, 4.6*50 mm, 3pm, Gradient: 30% B in 18 min, 220 nm.

[0417] The separated enantiomers were subjected to anaiytical chiral HPLC analysis (Instrument Name: Shimadzu LC-20AD; Mobile Phase A: n-Hexane; Mobile Phase B: Ethanol; Column; CHIRALPAK IC-3, 50*4.6mm, 3um IC30CC-SC002). This resulted in Compound 39h, Diastereomer 1 (chiral-HPLC analysis conditions: Rt = 2.03 min) and Compound 39h, Diastereomer 2 (chiral-HPLC analysis conditions: Rt = 2.89 min). LCMS (ES) [M+l]⁺ m/z: 440

Step 9a

[0418] Into a 25-mL vial, was placed Compound 39h, Diastereomer 1 (288 mg, 0.66 mmol, 1.0 equiv), CH3CN (8.0 mL), and Yb(OTf)i (203 mg, 0.33 mmol, 0.50 equiv). The mixture was stirred for 2 h at room température. The reaction solution was directly purified by Prep-HPLC with the following conditions (SHIMADZU (HPLC-01): Column, XBridge Cl8 OBD Prep

Column, ΙΟμιη, 19 mm X 250 mm, mobile phase, Water (0.1% FA) and CH3CN (5% Phase B up to 50% in 1.2 min), Detector, UV 254 nm. This resulted in Compound 34, Diastereomer 1. LCMS (ES, m/z): [M+H]⁺: 400. Ή-NMR (300 MHz, DMSO-ώ,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).

Step 9b

[0419] Into a 25-mL round-bottom flask, was placed Compound 39h, Diastereomer 2 (307 mg, 0.70 mmol, 1.0 equiv), CH3CN (8.0 mL), Yb(OTf)₃ (203 mg, 0.35 mmol, 0.50 equiv). The reaction solution was stirred for 2 h at room température. The reaction solution was directly purified by Prep-HPLC with the following conditions (SHIMADZU (HPLC-01): Column,

XBridge Cl8 OBD Prep Column, 10 μπι, 19 mm X 250 mm, mobile phase, Water (0.1%FA) and CH3CN (5% Phase B up to 50% in 12 min), Detector, UV 254 nm. Compound 34,

I

166

Diastereomer 2 was obtained. LCMS (ES, m/z): [M+H]⁺; 400. ^-NMR (300 MHz, DMSO-îZô, 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), 5.254.56 (m, 6H), 3.47-2.88 (m, 4H), 1.94-1.37 (m, 6H).

Example 40. 2-hydroxy-6-{[(3S)-4-{2-[(lS)-l-hydroxyethyl]pyridine-35 carbonyl}morpholin-3-yl]methoxy}benzaldehyde and 2-hydroxy-6-{[(3S)-4-{2-[(lR)-lhydroxyethyl]pyridine-3-carbonyl}morpholîn-3-yl]methoxy}benzaldehyde

[0420] Compound 41, Diastereomer 1 and Compound 41, Diastereomer 2 were synthesized according to Scheme 40.

o

40a

Scheme 40

Compound 40g

Diastereomer 1 and Diasteromer 2

Step 7a or Step 7b

Compound 41

Diastereomer 1 and Diasteromer 2

167

Step 1

[0421] Into a 250-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed a mixture of 3-bromopyridine-2-carbaldehyde (10.0 g, 53.7 mmol, 1.00 equiv), tctrahydrofuran (150 mL), and bromo(methyl)magnesium (35.8 mL, 2.0 5 equiv) was dropwised at -78 °C. The solution was stirred for 30 minutes at -78 °C and then allowed to room température over 30 minutes. The reaction was then quenched by the addition of 100 mL. The resulting solution was extracted with 3x100 mL of ethyl acetate. The resulting mixture was washed with 1 x 100 mL of brine. The mixture was dried over anhydrous sodium sulfate and concentrated. This resulted in l-(3-bromopyridin-2-yI)ethanol. LCMS (ES) [M+l]⁺ m/z: 202.

Step 2

[0422] Into a 250-mL round-bottom flask, was placed a mixture of l-(3-bromopyridîn-2yl)ethanol (8.00 g, 39.5 mmol, 1.00 equiv), DMF (80.0 mL), tert-butyl(chloro)diphenylsilane (16.3 g, 59.3 mmol, 1.50 equiv), and imîdazole (5.39 g, 79.1 mmol, 2.00 equiv). The resulting 15 solution was stirred for 16 hours at room température. The reaction was then quenched by the addition of 500 mL of water. The resulting solution was extracted with 3x150 mL of ethyl acetate. The resulting mixture was washed with 1 xl50 mL of brine. The mixture was dried over anhydrous sodium sulfate and concentrated. The residue was applîed onto a silica gel column with ethyl acetate/petroleum ether (1/9). This resulted in 3-bromo-2-[l-[(tert20 butyldiphenylsilyl)oxy]ethyl]pyridine. LCMS (ES) [M+l] ⁺ m/z: 440.1.

Step 3

[0423] Into a 1000 mL pressure tank reactor, was placed a mixture of 3-bromo-2-[l-[(tertbutyldiphenylsilyl)oxy]ethyl]pyridine (14.0 g, 31.7 mmol, 1.00 equiv), methanoi (200 mL), Et₃N (6.43 g, 63.5 mmol, 2.00 equiv), and Pd(dppf)Cl₂ (2.33 g, 3.18 mmol, 0.10 equiv). The reactor 25 was evacuated and flushed three times with nitrogen, followed by flushing 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 with ethyl acetate/petroleum ether (2/23) as eluent. This resulted in methyl 2-[l-[(terl-butyldiphenylsilyl)oxy]ethyl]pyridine3-carboxylate. LCMS (ES) [M+l] ⁺ m/z: 420.2.

168

Step 4

[0424] into a 250-mL round-bottom flask, was placed a mixture of methyl 2-[l-[(tertbutyldiphenylsilyl)oxy]ethyl]pyridine-3-carboxylate (10.0 g, 23.8 mmol, 1.00 equiv), MeOH (100 mL), and LiOH (1.71 g, 71.4 mmol, 3.00 equiv). The resulting solution was stirred for 3 5 hours at 50 degrees C. The resulting mixture was concentrated. The resulting solution was diluted with 100 mL of H₂O. The pH value of the solution was adjusted to 3 with HCl (2mol/L). The solids were coilected by filtration. This resulted in 2-[l-[(tertbutyldiphenylsilyl)oxy]ethyl]pyridine-3-carboxylic acid. LCMS (ES) [M+l] ⁺ m/z: 406.2.

Step 5

[0425] Into a 250-mL round-bottom flask, was placed a mixture of 2-[l-[(tertbutyldiphenylsilyl)oxy]ethyl]pyrîdine-3-carboxylic acid (7.00 g, 17.6 mmol, 1.00 equiv), DCM (100 mL), (3R)-morpholin-3-ylmethanol hydrochloride (3.45 g, 22.4 mmol, 1.30 equiv), DIEA (6.69 g, 51.7 mmol, 3.0 equiv), and HATU (7.88 g, 20.7 mmol, 1.2 equiv). The resulting solution was stirred for 3 hours at room température. The resulting mixture was concentrated.

The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (3/2) as eiuent. This resulted in [(3R)-4-(2-[l-[(tert-butyldiphenylsilyl)oxy]ethyl]pyridine3-carbonyl)morpholin-3-yl]methanol. LCMS (ES) [M+l] ⁺ m/z: 505.3.

Step 6

[0426] Into a 250-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen, was placed a mixture of [(3R)-4-[2-[(lS)-l-[(lertbutyldiphenylsilyl)oxy]ethyl]pyridine-3-carbonyl]morpholin-3-yl]methanol (2.0 g, 3.96 mmol, 1.00 equiv), DCM (100 mL), 2,6-dihydroxybenzaldehyde (0.71 g, 5.15 mmol, 1.30 equiv), and PPh? (1.56 g, 5.94 mmol, 1.50 equiv). DBAD (1.00 g, 4.35 mmol, 1.10 equiv) was added dropwise at 0 °C. The resulting solution was stirred for 16 hours at room température. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1/1) as eluents.

[0427] This resulted in Compound 40g, Diastereomer 1 (LCMS, Rétention time: 1.896 min), and Compound 40g, Diastereomer 2 (LCMS rétention time : 1.872 min, (ES) [M+l] ⁺ m/z;

625.2). LCMS analysis conditions: Instrument: Shimadzu LC2020; Column: Kinetex XB-C18, 30 50*3.0 mm, Particle sîze 2.6 um; Mobile phase A: Water/0.05%TFA; Mobile phase B:

Acetonitrile/0.05%TFA; Gradient: 5-100% B in 3 min.

169

Step 7A

[0428] Into a 20-mL vial, was placed a solution of Compound 40g, Diastereomer 1 (400 mg, 0.640 mmol, 1.00 equiv), THF (4.00 mL), and TBAF/THF (3.21 mL, 3.20 mmol, 5.00 equiv). The resulting solution was stirred for 3 hours at 45 degrees C. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (99/l~l/9).The crude reaction mixture was filtered and subjected to reverse phase préparative HPLC (XB-C18, 50-250 mm,10 mM; gradient elution of 10% MeCN in water to 45% MeCN în water over a 20 min period, where both solvents contain 0.1% formic acid), and the product was analyzed by analytical chiral HPLC (Instrument: Shimadzu LC-20AD; Mobile Phase A: n-Hexane(0.1%TFA); Mobile Phase

B:EtOH/MeOH =1/1; Conc. of Phase B: 20.0%; Column: CHIRALPAK IC-3, 50*4.6mm, 3um 1C30CC-SC002; Column ID: AY30CC-SK001; Flow Rate: 1.000 mL/min). This resulted in Compound 41, Diastereomer 1. Analytical chiral HPLC Rétention time = 5.801 min. LCMS (ES) [M+l] ⁺ m/z: 387.1. *H NMR (300 MHz, DMSO-de) 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).

Step 7B

[0429] Into a 20-mL vial, was placed a solution of Compound 40g, Diastereomer 2 (500 mg, 0.800 mmol, 1.00 equiv), THF (5.00 mL), and TBAF (4.01 mL, 4.00 mmol, 5.00 equiv). The resulting solution was stirred for 3 hours at 45 degrees C. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (99/1-1/9). The crude reaction mixture was filtered and subjected to reverse phase préparative HPLC (XB-C18? 50-250 mm, 10 mM; gradient elution of 10% MeCN in water to 45% MeCN in water over a 20 min period, where both solvents contain 0.1% formic acid), and the product was analyzed by analytical chiral HPLC (Instrument: Shimadzu LC-20AD; Mobile Phase A: n-Hexane(0.1%TFA); Mobile Phase

B:EtOH/MeOH =1/1; Conc. of Phase B: 20.0%; Column: CHIRALPAK IC-3, 50*4.6mm, 3um IC30CC-SC002; Column ID: AY30CC-SK001; Flow Rate: 1.000 mL/min). This resulted in Compound 41, Diastereomer 2. Analytical chiral HPLC Rétention time = 4.128 min. LCMS (ES) [M+l] ⁺ m/z: 387.1. Ή NMR (300 MHz, DMSO-î/ô) δ 11.84-11.20 (br, 1H), 10.38-10.15 (m, 1H), 8.58-8.53 (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).

170

Example 41. 2-hydroxy-6-{[(3S)-4-{2-[(2S)-2-hydroxypropyl]pyridine-3carbonyl}morpholin-3-yI] methoxy} benzaldehy de and 2-hydroxy-6-{[(3S)-4-{2’[(2R)-2hy droxy propy 1] py ridine-3-ca rbony l}morphol in -3-y 11 methoxy }benzalde hy de

[0430] Compound 42, Diastereomer 1 and Compound 42, Diastereomer 2 were synthesized 5 according to Scheme 41.

Scheme 41

O Br L I ----- ί SteP 1 41a N Step 4 A OH LI Step 6

0^ γ । ----- [ Y i ----- il A NAYqh Step 2 OTBDPS St®P 3 M'A 41b 41c '^'OTBDPS 41d \ OTBDPS G j HCI \,OTBDPS Y N ï < o Y ____ü_______D T ' j Y,0H Step 5 lA'A O ° ^OH 41e 41f γΟΤΒϋΡδ ^OTBDPS il O A ° S 0 k ArCHO J^cho kA f T ^OH aaoh Compound 41g Diastereomer 1 and Diastereomer 2 I I Step 7a or Step 7b ',,,.γΗ κγπ ArY · ArY 0 A 0 A J^ChiO Y^CHO Y'QH Ύχ οη

Compound 42

Diastereomer 1 and Diastereomer 2

171

Step 1

[0431] Into a 1000-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of nitrogen was placed 3-bromo-2-methylpyridine (25 g, 145.33 mmol, 1.00 equiv) and THF (500.00 mL). This was followed by lhe addition of LDA (87.20 mL, 174.40 mmol,

1.20 equiv) dropwise with stirring at -78 °C. The resulting solution was stirred for 1 h at -78 °C.

To this mixture was added acetaldehyde (7.04 g, 159.81 mmol, 1.10 equiv) dropwise with stirring at -78 °C. The resulting solution was stirred for 1 h at -78 °C. The reaction was then quenched by the addition of 300 mL of saturated NH4CI solution. The resulting mixture was extracted with 3x300 mL of ethyl acetate, and the organic layers were combined, dried over lü anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography using THF/PE (20%) as eluent to yield l-(3-bromopyridin-2-yl)propan-2-oI. LCMS (ES) [M+l]⁺ m/z: 216.

Step 2

[0432] Into a 1000-mL round-bottom flask, was placed l-(3-bromopyridin-2-yI)piOpan-2-o] (15.00 g, 69.42 mmol, 1.00 equiv), imidazole (9.45 g, 138.81 mmol, 2.00 equiv), DMF (300.00 mL), DMAP (0.85 g, 6.94 mmol, 0.1 equiv) and TBDPSC1 (22.90 g, 83.30 mmol, 1.20 equiv). The resulting solution was stirred for overnight at 60 °C. The reaction mixture was cooled to room température. The reaction was then quenched by the addition of 300 mL of water. The resulting solution was extracted with 3x300 mL of ethyl acetate, and the organic layers were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography using THF/PE (5%) to yield 3-bromo-2-[2-[(tertbutyldiphenylsilyl)oxy]propyl]pyridine. LCMS (ES) [M+l]⁺ m/z: 454.

Step 3

[0433] Into a 2000-mL pressure tank reactor was placed 3-bromo-2-[2-[(tert25 butyldiphenylsîlyl)oxy]propyl]pyridine (25.00 g, 55.00 mmol, 1.00 equiv), MeOH (800.00 mL), TEA (11.13 g, 110.01 mmol, 2.00 equiv), and Pd(dppf)Ch (4.02 g, 5.49 mmol, 0.10 equiv). The reactor was evacuated and flushed three times with nitrogen, followed by flushing with 30 atm CO. The resulting solution was stirred for overnight at 110 °C. The reaction mixture was cooled to room température. The resulting mixture was concentrated. The residue was purified with silica gel column chromatography using THF/PE (7%) to yield methyl 2-[2-[(tertbutyldiphenylsilyl)oxy]propyl]pyridine-3-carboxylate. LCMS (ES) [M+l]⁺ m/z: 434.

172

Step 4

[0434] Into a 1000-mL round-bottom flask, was placed methyl 2-[2-[(tertbutyldiphenylsily])oxy]propyl]pyridine-3-carboxylate (20.00 g, 46.12 mmol, 1.00 equiv), MeOH (400 mL), H₂O (200 mL), and L1OHH2O (3.87 g, 92.22 mmol, 2.00 equiv). The resulting solution was stirred for 4 h at 50 °C. The reaction mixture was cooled to room température. The resulting mixture was concentrated. The resulting solution was extracted with 200 mL of ethyl acetate, and the aqueous layers combined. The pH value of the solution was adjusted to 4-5 with HCl (1 mol/L). The resulting précipitâtes were collected by filtration and dried under infrared light. This resulted in 2-[2-[(teri-butyldiphenyIsilyl)oxy]propyl]pyridine-3-carboxylic acid.

LCMS (ES) [M+l]⁺ m/z: 420.

Step 5

[0435] Into a 250-mL 3-necked round-bottom flask, was placed 2-[2-[(tertbutyldiphenylsilyl)oxy]propyl]pyridine-3-carboxylic acid (4.00 g, 9.53 mmol, 1.00 equiv), (3R)morphoIin-3-ylmethanol hydrochloride (1.76 g, 11.46 mmol, 1.20 equiv), DCM (100.00 mL), 15 and DIEA (3.70 g, 28.59 mmol, 3.00 equiv). This was followed by the addition of HATU (4.35 g, 11.44 mmol, 1.20 equiv) in portions at 0 °C. The resulting solution was stirred for 3 h at room température. The reaction was then quenched by the addition of 100 mL of water. The resulting solution was extracted with 3x100 mL of dichloromethane, and the organic layers were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified with silica gel column chromatography using THF/PE (25%) as eluent to yield [(3R)-4-(2-[2-[(tertbutyldiphenylsilyl)oxy]propyl]pyridine-3-carbonyl)morpholin-3-yl]methanol. LCMS (ES) [M+l]⁺ m/z: 519.

Step 6

[0436] Into a 250-mL 3-necked round-bottom flask, was placed 2,6-dihydroxybenzaldehyde (0.96 g, 6.94 mmol, 1.20 equiv), [(3R)-4-(2-[2-[(tert-butyldiphenyIsilyl)oxy]propyl]pyridine-3carbonyl)morpholin-3-yl]methanol (3.00 g, 5.78 mmol, 1.00 equiv), PPh₃ (1.82 g, 6.94 mmol, 1.20 equiv), and DCM (100.00 mL). This was followed by the addition of DIAD (1.40 g, 6.92 mmol, 1.2Ü equiv) dropwîse with stirring at 0 °C. The resulting solution was stirred for overnight at room température. The resulting mixture was concentrated. The residue was purified with silica gel column usîng THF/PE (30%) as eluent to give crude product. The crude product was purified by Prep-HPLC with the followîng conditions: Column. Welch XB-C18 50*250mm, lOum, mobile phase, Water (0.1% TFA) and ACN (50% in 100 min); Detector,

173

254. This resulted in Compound 41g, Diastereomer 1 (Rétention time = 70 min) and Compound 41g, Diastereomer 2 (Rétention time = 9Ü min). LCMS (ES) [M+l]⁺ m/z: 639.

Step 7A

[0437] Into a 40-mL vial, was placed Compound 41g, Diastereomer 1 (1.2 g, 1.88 mmol, 1.00 equiv), THF (9.00 mL), and TBAF/THF (9.39 mL, 9.39 mmol, 5.00 equiv). The resulting solution was stirred for 5 h at 45 °C. The reaction mixture was cooled to room température. The crude product was purified by Prep-HPLC with the following conditions: Column, XBridge Prep Cl8 OBD Column, 19cm, 150mm, 5um; mobile phase, Water (0.1% HCOOH) and ACN (5% to 55% in 15 min); Detector, 254. This resulted in Compound 42, Diastereomer 1. The final compound was analyzed with Chiral HPLC with these conditions: Instrument: SHIMADZU LC20AT; Mobile Phase A: n-Hexane; Mobile Phase B: Mobile Phase B; Conc. of Phase B: 50.0%, Flow Rate Column: 1.000 mL/min: CHIRALPAK AY-3, 4.6*50 mm, 3pm; Column ID: AY30CC-SK001; Rétention time = 3.35 min. LCMS: (ES, m/z): [M+H]*: 401. ^lH-NMR (300 MHz, DMSO-ikppm): δ 1 1.80-11.67 (m, 1H), 10.34-10.23 (m, 1H), 8.58 (dd, J = 4.9, 1.7 Hz, 15 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).

Step 7B

[0438] Into a 40-mL vial, was placed Compound 41g, Diastereomer 2 (1.20 g, 1.88 mmol, 1.00 20 equiv), THF (9.0Ü mL), and TBAF (9.39 mL, 9.39 mmol, 5.00 equiv). The resulting solution was stirred for 5 h at 45 °C. The reaction mixture was cooled to room température. The crude product was purified by Prep-HPLC with the following conditions: Column, XBridge Prep C18 OBD Column, 19cm, 150mm, 5um; mobile phase, Water (0.1% HCOOH) and ACN (5% to 55% in 15 min); Detector, 254. This resulted in Compound 42, Diastereomer 2. The final compound 25 was analyzed with Chiral HPLC with these conditions: Instrument: SHIMADZU LC-20AT;

Mobile Phase A: a-Hexane; Mobile Phase B: Mobile Phase B; Conc. of Phase B: 50.0%; Flow Rate Column: 1.000 mL/min: CHIRALPAK AY-3, 4.6*50 mm, 3gm; Column 1D: AY30CCSK001; Rétention time = 1.91 min. LCMS: (ES, m/z): [M+H]⁺: 401. ’H-NMR: (300 MHz, DMS0-d₆, ppm): δ 11.82-11.69 (m, 1H), 10.33-10.23 (m, 1H), 8.58 (dd, J = 4.9, 1.8 Hz, 1H), 30 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).

174

Example 42. 2-hydroxy-6-{[(3R)-4-{2-[(lS)-l-hycIroxyethyl]pyridine-3 carbonyl[thiomorpholin-3-yl]methoxy [benzaldehyde and 2-hydroxy-6-{[(3R)-4-{2-[(lR)-lhydroxyethyl] pyridine-3-carbon y I [thiomorpholi n-3-yl] methoxy [benzaldehyde

[0439] Compound 43, Diastereomer 1 and Compound 43, Diastereomer 2 were synthesized according to Scheme 42.

Scheme 42

Compound 43

Diastereomer 1 and Diastereomer 2

Step 1

[0440] To a solution of 2-[l-[(tert-butyldiphenylsilyl)oxy]ethyl]pyridine-3-carboxyiic acid (2.0 10 g, 4.93 mmol, 1.0 equiv)) in DMF (20.0 mL) was added DIPEA (1.27 g, 9.8 mmol, 2.0 equiv) and HATU (2.25 g, 5.9 mmol, 1.2 equiv) at 0 °C. After the reaction mixture was stirred at 0 °C for 20 min, (3R)-thiomorpholin-3-ylmethanol (720 mg, 5.42 mmol, 1.10 equiv) was added in portions. The resulting solution was stirred for 2 h at 25 °C. The reaction was then quenched by the addition of 50 mL of water. The resulting solution was extracted with 3x80 mL of ethyl acetate. The combined organîc layer was 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. This resulted in [(3R)-4-(2-[l-[(tertbutyldiphenylsilyl)oxy]ethyl]pyridine-3-carbonyl)thiomorpholin-3-yl]methanoL LCMS (ES) [M+l] ⁺ m/z: 521.2; Ή-NMR: (300 MHz, DMSO-d6, ppm): δ 8.75 (d, J =5.4 Hz, 1H), 7.96 (s, 20 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

175 * (m, 1H), 2.74-2.70 (m, 1H), 2.38-2.33 (m, 1H), 1.79-1.69 (m, 1H), 1.59-1.10 (ni, 3H), 0.92 (s,

9H).

Step 2

[0441] A solution of [(3R)-4-(2-[l-[(tert-butyldiphenyIsilyl)oxy]ethyl]pyridine-35 carbonyl)thiomorpholin-3-yl]methanol (1.3 g, 2.5 mmol, 1.0 equiv), 2,6-dihydroxybenzaldehyde (380 mg, 2.72 mmol, 1.1 equiv) and PPh₃ (980 mg, 3.75 mmol, 1.5 equiv) in DCM (200 mL) was cooled lo 0 °C under Argon atmosphère. A solution of DBAD (690 mg, 3.0 mmol, 1.2 equiv) in DCM (30 mL) was added dropwise. After the addition, the resulting solution was stirred for 16 h at 0-25 °C. The reaction was concentrated under vacuum. The residue was purified by silica gel column using ethyl acetate/petroleum ether (1:100 to 1:5) as eluent. This resulted in 2-[[(3R)-4-(2-[l-[(terl-butyldiphenylsilyl)oxy]ethyl]pyridine-3carbonyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde. LCMS (ES) [M+l] ⁺ m/z: 641.2; ’H-NMR (300 MHz, CDCh, ppm): δ 11.96 (s, 1H), 10.25 (br, 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).

Step 3

[0442] Into a 100-mL 3-necked round-bottom flask, was placed 2-[[(3R)-4-(2-[l-[(tertbutyldiphenylsilyl)oxy]ethyl]pyridine-3-carbonyl)thiomorpholin-3-yl]methoxy]-6hydroxybenzaldehyde (2.0 g, 3.12 mmol, 1.0 equiv) and THF (10 mL). After the reaction was cooled to 0 °C, TB AF (1.63 g, 6.24 mmol, 2.0 equiv) was added in portions. The resulting solution was stirred for 5 h at 45 °C. The resulting solution was quenched with NH4CI (20 mL, 2N) and extracted with ethyl acetate (50 mLx3); the organic layers were combined and concentrated. The residue was purified by silica gel column eluted with PE/EA= 100:1 to 1:1 to give the racemate product, which was purified by Préparative Chiral-HPLC with the followîng conditions: Co1umn:Lux Amylose-1, 50*250mm, lOum; Mobile phase : A:n-Hexane B:Ethanol; Flow rate: 90 mUmin; Gradient:50%B in 50min; 220nm. The isolated diastereomers were analyzed by analytical HPLC using the followîng conditions: Instrument: SHIMADZU LC20AT; Mobile Phase A: n-Hexane; Mobile Phase B: Ethanol; Conc. of Phase B: 50.0%; Flow Rate: 1.000 mL/min; Column; Lux Amylose-1,4.6*100 mm, 3pm; Column ID: H18-344853.

This resulted in Compound 43, Diastereomer 1 and Compound 43, Diastereomer 2.

[0443] Data for Compound 43, Diastereomer 1 : Chiral HPLC rétention time 8.31 min; LCMS (ES, m/z); [M+H] ⁺: 403.1; Ή-NMR: (300 MHz, DMSO-t/6, ppm): δ H.85 (br, 1H), 10.3520663

176

10.16 (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).

[0444] Data for Compound 43, Diastereomer 2: Chiral HPLC rétention time 5.30 min; LCMS (ES, m/z): [M+H] ⁺: 403.1; Ή-NMR: (300 MHz, DMSO-r/6, ppm): δ 1 1.79 (br, 1H), 10.325 10.16 (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. 2-hydiOxy-6-{[(3R)-4-{2-[(2S)-2-hydroxypropyl]pyridine-3carbonyI}thiomorpholin-3-yl]methoxy}benzaIdehydeand 2-hydroxy-6-{[(3R)-4-{2-[(2R)-2hydroxypropyl]pyridine-3-carbonyl}thiomorpholin-3-yl]methoxy}benzaldehyde

[0445] Compound 44, Diastereomer 1 and Compound 44, Diastereomer 2 were synthesized according to Scheme 43.

Scheme 43

OH

Step 3

CHO

OH

CHO

OH

Compound 44

Diastereomer 1 and 2

Step 1

[0446] To a solution of 2-[2-[(tert-butyldiphenylsilyl)oxy]propyl]pyridine-3-carboxyïic acid (2.0 g, 4.76 mmol, 1.0 equiv) in DMF (20.0 mL) was added DIPEA (1.23 g, 9.5 mmol, 2 equiv) and HATU (2.17 g, 5.720 mmol, 1.20 equiv) at 0°C. After the reaction mixture was stirred at 0

177 °C for 20 min, (3R)-thiomorpholin-3-ylmethanol (0.70 g, 5.243 mmol, 1.1 equiv) was added in portions. The resulting solution was stirred for 2 h at 25 °C. The reaction was then quenched by the addition of 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 5 purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:100 to 1:1) as eluent. This resulted in [(3R)-4-(2-[2-[(tert-butyIdiphenyIsilyl)oxy]propyi]pyridine-3carbonyI)thiomorpholin-3-yl]methanol. LCMS (ES) [M+l] ⁺ m/z: 535.2.

Step 2

[0447] Into a 250-mL 3-necked round-bottom flask purged and maintained with an inert atmosphère of argon, was placed [(3R)-4-(2-[2-[(tert-butyIdiphenylsilyl)oxy]propyl]pyridine-3carbonyl)thiomorpholin-3-yl]methanol (1.5 g, 2.8 mmol, 1.0 equiv), 2,6-dihydroxybenzaldehyde (Ü.43 g, 3.1 mmol, 1.1 equiv), DCM (150.00 mL) and PPh₃ (1.1 g, 4.2 mmol, 1.5 equiv). After the reaction was cooled to 0 °C, a solution of DBAD (0.78 g, 3.36 mmol, 1.2 equiv) in DCM (10 mL) was added dropwise. The resulting solution was stirred for 16 h at 0 to 25 ^ÛC. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography with ethyl acetate/petroleum ether (1:100 to 1:5) as eluent. This resulted in 1[[(3R)-4-(2-[2-[(tert-butyldiphenylsiIyl)oxy]propyl]pyridine-3-carbonyl)thiomorpholin-3yl]methoxy]-6-hydroxybenzaldehyde. LCMS (ES) [M+l] ⁺ m/z: 655.2; *H-NMR: (300 MHz, DMSO-d6, ppm): δ 11.76 (br, 1H), 10.39 (br, 1H), 8.54-8.48 (m, 1H), 7.86-7.27(m, 13H), 6.7820 6.55 (m, 2H), 5.41 (br, 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).

Step 3

[0448] Into a 100-mL 3-necked round-bottom flask, was placed 2-[[(3R)-4-(2-[2-[(tertbutyldiphenylsilyl)oxy]propyl]pyridine-3-carbonyI)lhiomorphoIm-3-yl]methoxy]-625 hydroxybenzaldehyde (LO g, 1.52 mmol, LO eq.) and THF (10 mL). After the réaction was cooled to 0 °C, a solution of TEA 3HF (1.0 g, 80.9 mmol, 3.0 equiv) was added dropwise. The resulting solution was stirred for 5 h at 45 °C. The pH value of the solution was adjusted to 8 with NaHCCh (2 mol/L). The reaction was extracted with ethyl acetate (50 mLx3), and the organic layers combined and concentrated. The crude product was purified by silica gel column chromatography eluted with PE/EA= 100:1 to 1:1 to give the racemate product.

[0449] The racemate product was purified by Chiral-HPLC (Conditions: Column:Lux Amylose-1, 50*250mm, lOum; Mobile phase: A:n-Hexane B:Ethanol; Flow rate: 90 mL/min;

¹⁷⁸

Gradient:50%B in 36mîn; 220nm) and was analyzed by analytical HPLC (Conditions: Instrument: SHIMADZU LC-20AT; Mobile Phase A: n-Hexane; Mobile Phase B: Ethanol;

Conc. of Phase B: 50.0%; Flow Rate: 1.000 mL/min; Column: Lux Amylose-l, 4.6*100 mm, 3pm; Column ID: H18-344853). This resulted in Compound 44, Diastereomer 1 and Compound 5 44, Diastereomer 2.

[0450] Data for Compound 44, Diastereomer 1: Chiral HPLC rétention time = 4.85 min; LCMS (ES, m/z): [M+H] ⁺: 417.2; ¹H-NMR: (300 MHz, DMSO-J6, ppm): S 11.77 (br, 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).

[0451] Data for Compound 44, Diastereomer 2: Chiral HPLC rétention time 6.94 min; LCMS (ES, m/z): [M+H] ⁺: 417.2; iH-NMR: (300 MHz, DMSO-76, 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).

[0452] Compounds 6-9, and 18 in Table 3 were synthesized according to the methods described herein and appropriate modifications thereof.

Table 3

Compound Number______	Structure	Mass Spectrometry Data
6	.OH 0 O ^^OH	384.1 (MH+)
7	Z z o=? —\ z . O / \ T O \=o 1 T	386.2 (MH+)

179

Compound Number	Structure	Mass Spectrometry Data
8	y^ T 0 \ P 0—\ y° X \ O —z \\ Go Z /7 \\ //	387.1 (MH+)
9	\ Z o^/y A _- —/ 1 0 0 >0 Z	371.1 (MH+)
18	OH cG 0 ^0 0 G	LCMS (ES) [M+l]+ m/z 399.2

Biological Assays

[0453] Whole blood assay: Oxygen equilibrium curves (OECs) were collected using a TCS Hemox Analyzcr (TCS Scientific Company, New Hope, PA, USA) to measure changes in the binding affinity of O₂ to Hb. Whole blood was incubated for 1 h at 37 C with the îndicated compounds in an equimolar ratio of hemoglobin to compound and diluted into TES (2-[[l,3dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]ethanesulfonic acid)/saline buffer prior to measurements. For example, for whole blood at 20% hematocrit [Hct], which corresponds to 1 mM Hb, a compound concentration of 1 mM was used (for example, for compounds 1-5), and the incubated sample diluted 50- to 100-fold. The concentration for compounds 6-44 (Diastereomers 1 and 2) varied but remaîned in equimolar ratio to hemoglobin. The diluted samples were then oxygenated with compressed air within the Hemox Analyzer and the OECs

180

I were collected during deoxygenation as previously described (Guarnone et aL, Haematologica, 1995, 80, 426-430). p50 (partial pressure of O₂ at which Hb is 50% saturated with O₂) values were obtained using a non-linear régression analysis. Percentage change in p50 [Δρ50 (%)] was calculated as follows: Δρ50 (%) = [(p50 ofcontrol)~p50 with compound)/p50 control] x 100.

Resulting data is shown in Table 4. Enantiomer 1 and Enantiomer 2 of Compound 13 also exhibit a Δρ50 of about 61.0% to about 80.6%.

Table 4

Compound 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
11	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
30	74.19
31	74.03
32	60.55
33	49.03
34	79.46_________________

181

Compound Number	Delta-p50 (%)
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

[0454] CYP (PXR) Assay: PXR nuclear receptor activation utilizing stably-transfected human hepaloma cell Unes (DPX2) were seeded in a 96-well plate. Twenty-four hours after seeding, the cells were treated with selected concentrations of compounds in duplicate wells, and cells then returned to the incubator for an additional 24 h. At the end of this incubation period, the number of viable cells/well were determined using Promega’s Cell Titer Fluor cytotoxicity assay. Subsequently, Promega’s ONE-Glo were added to the same wells and reporter gene activity were assessed. The average luminescent units for each compound dose of two replicates were divided by the average for the DMSO solvent control to détermine the fold-induction. In 10 accordance with industry standard, a threshold of > 2.5-fold was used to flag compounds that hâve în vivo CYP induction risk.

[0455] Structures of reference compounds (Compound A, Compound B, and Compound C) are shown below in Table 5.

182

Table 5

	Reference Compound A	Reference Compound B	Reference Compound C
Structure	0 O ôC	F x 0 \ /° /o-^/oÂ7 \—Z ^0	.N. / cçç 0 Ό 0

[0456] Results for various compounds disclosed herein and select reference compounds are summarized in Table 6.

Table 6

Compound	CYP (PXR) Flag
Reference Compound A	Y
Reference Compound B	Y
Reference Compound C	Y1
J_________________	N1
8	N
12	N
Compound 13 (Enantiomer 1)	N_______

CYP (PXR) Flag based on fold PXR activation (human, at 30 μΜ):

Y, PXR activation > 2.5-fold;

N, PXR activation < 2.5-fold.

¹ al 25 μΜ.

[0457] Rat PK: A group of fasted male Sprague-Dawley rats were dosed via oral gavage at 10 mg/kg with test articles formulated in 0.5% methylcellulose suspension. Blood samples were collected through jugular vein at pre-selected time points. Blood samples were prepared by protein précipitation with ACN, vortexed and then centrifuged before supernatants were transferred for bîoanalysis. Test article concentrations were measured by HPLC-MS-MS. Pharmacokinetic parameters were calculated using non-compartment analysis. The blood/plasma ratio was calculated by dividing the AUC|_HSt (i.e., the area under the curve calculated from t = 0 to the last détectable time-point) in blood by the AUCiast in plasma. The T1/2 was calculated via a linear régression of the terminal phase of the blood-time concentration profile.

183

[0458] Results for various compounds disclosed herein and select référencé compounds (Compound A and Compound B) are summarized in Table 7.

Table 7

Compound	Tl/2 (h)	Blood/Plasma ratio
Référencé Compound A	29	75
Reference Compound B	29.8	98
1	58	162
8	69	105
10 (Enantiomer 2)	112	212
11	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

>k ψ ik

[0459] Unless otherwise defined, ali technical and scientific terms used herein hâve the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0460] The inventions illustratively described herein may suitably be practiced in the absence of any element or éléments, limitation or limitations, not specifically disclosed herein. Thus, for 10 example, the terms “comprising”, “including,” “containing”, etc. shah be read expansively and without limitation. Additionally, the terms and expressions employed herein hâve been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any équivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed.

184

[0461] Ail publications, patent applications, patents, and other references mentioned herein are expressly incorporated by reference in their entirety, to the same extent as if each were incorporated by reference indîvidually. In case of conflict, the présent spécification, including définitions, wîll control.

Claims (42)

1. CLAIMS:

   I. A compound of formula I

   or an isotopicaily enriched analog, stereoisomer, or mixture of stereoisomers thereof, or a pharmaceutically acceptable sait of each thereof, wherein:

   X is CH or N;

   Y is CH or N;

   Z is absent, CH₂, O, or S; and

   R¹ is mono-hydroxy-(CM alkyl), di-hydroxy-(Ci-4 alkyl), -CH2CH2OCH3, -CH2CH2CN,
2. 2. The compound of claim I, wherein X is CH.
3. 3. The compound of claim 1, wherein X is N.
4. 4. The compound of any one of the preceding daims, wherein Y is CH.
5. 5. The compound of any one of daims 1 -3, wherein Y is N.
6. 6. The compound of any one of the preceding daims, wherein Z is absent, CH₂, or O.
7. 7. The compound of any one of the preceding daims, wherein Z is absent.
8. 8. The compound of any one of daims 1-5, wherein Z is CH2.
9. 9. The compound of any one of daims 1-5, wherein Z is O.

   186
10. 10. The compound of any one of daims 1 -5, wherein Z is S.
11. 11. The compound of daim 1, of formula la:
12. 12. The compound of any one of the preceding daims, wherein R¹ is -CH₂OH, -CH2CH2OH,

    -CH2CH2CN, or
13. 13. The compound of any one of lhe preceding daims, wherein R¹ is -CH2OH or

    10 -CH2CH2OH.
14. 14. The compound of any one of daims 1-11, wherein R¹ is mono-hydroxy-(Ci^ alkyl) or di-hydroxy-(CM alkyl).

    15
15. 15. The compound of any one of daims I-11, wherein R¹ is hydroxym ethyl, 1 -hydroxyethyl,

    2-hydroxyethyl, 1,2-dîhydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, or 2-hydroxy-2methylpropyl.
16. 16. A compound selected from Table 1, or an isotopically enriched analog, stereoisomer, or 20 mixture of stereoîsomers thereof, or a pharmaceutically acceptable sait of each thereof.
17. 17. A compound selected from Table 1.
18. 18. A compound selected from Table 2, or an isotopically enriched analog, stereoisomer, or

    25 mixture of stereoîsomers thereof, or a pharmaceutically acceptable sait of each thereof.

    188 or an isotopically enriched analog, stereoisomer, or mixture of stereoisomers thereof, or a pharmaceutically acceptable sait of each thereof.
19. 20. A compound of formula:

    or a pharmaceutically acceptable sait of each thereof.

    10
20. 21. The compound of claim 20, wherein the compound is:

    or a pharmaceutically acceptable sait thereof.

    189
21. 22. The compound of claim 20, wherein the compound is:

    23.

    The compound of claim 20, wherein the compound is:

    5 or a pharmaceutically acceptable sait thereof.

    24.

    The compound of claim 20, wherein the compound is:

    25.

    The compound of claim 20, wherein the compound is:

    10 or a pharmaceutically acceptable sait thereof.

    190
22. 26. The compound of claim 20, wherein the compound is:
23. 27. The compound of claim 20, wherein tire compound is:

    OH

    5 or a pharmaceutically acceptable sait thereof.
24. 28. The compound of claim 20, wherein the compound is:

    OH

    29.

    The compound of claim 20, wherein the compound is:

    OH

    10 or a pharmaceutically acceptable sait thereof.

    191
25. 30. The compound of claim 20, wherein the compound is:

    OH
26. 31. The compound of claim 20, wherein the compound is:

    OH

    5 or a pharmaceutically acceptable sait thereof.
27. 32. The compound of claim 20, wherein the compound is:
28. 33. A pharmaceutical composition comprising a compound according to any one of the

    10 preceding daims, or an isotopically enriched analog, stereoisomer, or mixture of stereoisomers thereof, or a pharmaceutically acceptable sait of each thereof, and a pharmaceutically acceptable excipient.
29. 34. A pharmaceutical composition comprising a compound according to claim 17, and a

    15 pharmaceutically acceptable excipient.
30. 35. A compound according to any one of daims 1-32, or an isotopically enriched analog, stereoisomer, or mixture of stereoisomers thereof, or a pharmaceutically acceptable sait of each

    192 thereof, or a pharmaceutical composition of claim 33 for use in a method for increasing oxygen affïnity of hemoglobin S in a subject in need thereof.
31. 36. Use of a compound according to any one of daims 1-32, or an isotopically enriched analog, stereoisomer, or mixture of stereoisomers thereof, or a pharmaceutically acceptable sait of each thereof, in the manufacture of a pharmaceutical composition for increasing oxygen affïnity of hemoglobin S in a subject in need thereof.
32. 37. A compound according to claim 17 or a pharmaceutical composition of claim 34 for use in a method for increasing oxygen affïnity of hemoglobin S in a subject in need thereof.
33. 38. Use of a compound according to daim 17 in the manufacture of a pharmaceutical composition for increasing oxygen affïnity of hemoglobin S in a subject in need thereof.
34. 39. A compound according to any one of daims 1 -32, or an isotopically enriched analog, stereoisomer, or mixture of stereoisomers thereof, or a pharmaceutically acceptable sait of each thereof, or a pharmaceutical composition of claim 33 for use in a method for treating a disorder mediated by hemoglobin in a subject in need thereof.
35. 40. Use of a compound according to any one of daims 1 -32, or an isotopically enriched analog, stereoisomer, or mixture of stereoisomers thereof, or a pharmaceutically acceptable sait of each thereof, in the manufacture of a pharmaceutical composition for treating a disorder mediated by hemoglobin in a subject in need thereof.
36. 41. A compound according to daim 17 or a pharmaceutical composition of claim 34 for use in a method for treating a disorder mediated by hemoglobin in a subject in need thereof.
37. 42. Use of a compound according to daim 17 in the manufacture of a pharmaceutical composition for treating a disorder mediated by hemoglobin in a subject in need thereof.
38. 43. The compound for use or the use of any of daims 39 to 42, wherein the hemoglobin is sickle hemoglobin.
39. 44. A compound according to any one of daims 1-32, or an isotopically enriched analog, stereoisomer, or mixture of stereoisomers thereof, or a pharmaceutically acceptable sait of each

    193 thereof, or a pharmaceutical composition of claim 33 for use in a method for treating sickle cell disease in a subject in need thereof.
40. 45. Use of a compound according to any one of daims 1 -32, or an isotopically enriched

    5 analog, stereoisomer, or mixture of stereoisomers thereof, or a pharmaceutically acceptable sait of each thereof, in the manufacture of a pharmaceutical composition for treating sickle cell disease in a subject in need thereof.
41. 46. A compound according to daim 17 or a pharmaceutical composition of daim 34 for use 10 in a method for treating sickle cell disease in a subject in need thereof.
42. 47. Use of a compound according to daim 17 in the manufacture of a pharmaceutical composition for treating sickle cell disease in a subject in need thereof.