WO2022171110A1

WO2022171110A1 - Fused tricyclic compounds as rip1-kinase inhibitors and uses thereof

Info

Publication number: WO2022171110A1
Application number: PCT/CN2022/075608
Authority: WO
Inventors: Yuhua Ji; Yeyu Cao; Dawei Ma; Weiming He; Chao FANG; Jinlong ZHAO
Original assignee: Shanghai Institute of Organic Chemistry of CAS; Zai Lab US LLC
Current assignee: Shanghai Institute of Organic Chemistry of CAS; Zai Lab US LLC
Priority date: 2021-02-10
Filing date: 2022-02-09
Publication date: 2022-08-18
Anticipated expiration: 2023-08-10

Abstract

A compound of formula I below as RIP1 kinase inhibitor and uses thereof.

Description

FUSED TRICYCLIC COMPOUNDS AS RIP1-KINASE INHIBITORS AND USES THEREOF

This application claims the priority of the PCT/CN2021/076563, filed on February 10, 2021, the contents of which are incorporated herein by its entirety.

TECHNICAL FIELD

This application relates to compounds that inhibit cell necrotic and/or receptor-interacting protein-1 (RIP1) kinases and uses thereof.

BACKGROUND

RIP1 kinase has been found to be a central regulator of cell signaling, which functions in a variety of cellular pathways related to both cell survival and death. The cell death regulated by RIP1 kinase include apoptosis and necroptosis.

Studies have shown that RIP1 kinase inhibitors could provide therapeutic benefit in a subject with RIP1 kinase-mediated diseases or disorders, including but not limited to myocardial infarction, stroke, atherosclerosis, ischemia-reperfusion injury, inflammatory bowel diseases, psoriasis, rheumatoid arthritis, retinal degeneration and a number of other inflammatory diseases, hematological and solid organ malignancies, pancreatic cancer, bacterial infections and viral infections (including, but not limited to, tuberculosis and influenza) and Lysosomal storage diseases.

It is desired to have RIP1 kinase inhibitors with improved properties such as stability and pharmacokinetic profiles over existing RIP1 kinase inhibitors.

This application provides RIP1 kinase inhibitors that can show improved properties over certain existing compounds.

SUMMARY

Provided is a compound of formula I or a pharmaceutically acceptable salt thereof:

wherein

R ₁ is H, F, CN, CF ₃, or Cl,

R ₂ is H, -CH ₃, -CD ₃, or cyclopropyl,

A ring is a bicyclic fused ring system represented by

in which N ring is fused to the amide-containing seven-membered ring,

N ring is a pyridine, pyrazine, or phenyl optionally substituted with one or two groups selected from F and Cl,

M is a 5 or 6 membered heteroaryl or phenyl each of which is optionally substituted by one or two groups selected from oxo, Cl, -O-C _1-3 alkyl, OH, CN, C _1-3 alkyl, -S-C _1-3 alkyl, phenyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b,

W is independently selected from -C _1-3 alkyl, oxo, -OH, -COOC _1-3 alkyl, C _3-5 cycloalkyl, haloC _1-3 alkyl, -O-C _1-3 alkyl, Cl, and CN,

R _a is H, CH ₃, or CH ₂CH ₃,

R _b is selected from H, -C _1-3 alkyl, haloC _1-3 alkyl, -CH ₂-C _3-5 cycloclkyl, -CH ₂-phenyl, optionally substituted 5 or 6 membered heterocyclyl, C _3-5 cycloalkyl, - (CH ₂) _2-3OH, - (CH ₂) _2-3OCH ₃, wherein the optional substituent is selected from C _1-3 alkyl and C _3-5 cycloalkyl,

X is O or CH ₂,

B is

in which the amide nitrogen is attached to the amide-containing seven membered ring, X ₁ is CH or N, R _c is selected from H, -CH ₃, -CD ₃, -CH ₂CF ₃, -CH=CH ₂ _, and cyclopropyl, and R _c is attached to one of the nitrogen of the 5-membered ring,

L is C=O, CHCH ₃, C (CH ₃) ₂, O, CHOH, CH ₂ or

provided that when B is

N ring is a non-substituted phenyl, and (1) R _c is not cyclopropyl or -CH=CH ₂, (2) R ₂ is not cyclopropyl, and (3) L is not

then M ring is substituted by one or two groups selected from oxo, S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b excluding NH ₂, wherein W, R _a and R _b are as disclosed herein.

Also provided is a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

Also provided is a method of inhibiting RIP1-kinase in a subject in need thereof, comprising administering a therapeutically effective amount of a composition comprising the compound of formula I or a pharmaceutically acceptable salt thereof.

Also provided is a method of treating a disease or disorder mediated by RIP1-kinase in a subject in need thereof, comprising administering a therapeutically effective amount of a composition comprising the compound of formula I or a pharmaceutically acceptable salt thereof.

The disease or disorder mediated by RIP1-kinase include cancer and inflammatory diseases. For example, the disease or disorder include inflammatory bowel diseases, Crohn's disease, ulcerative colitis, psoriasis, retinal detachment, retinitis pigmentosa, macular degeneration, pancreatitis, atopic dermatitis, arthritis (including rheumatoid arthritis, osteoarthritis, spondylarthritis, gout, systemic onset juvenile idiopathic arthritis (SoJIA) , psoriatic arthritis) , systemic lupus erythematosus (SLE) , Sjogren's syndrome, systemic scleroderma, anti-phospholipid syndrome (APS) , vasculitis, liver damage/diseases (non-alcohol steatohepatitis, alcohol steatohepatitis, autoimmune hepatitis autoimmune hepatobiliary diseases, primary sclerosing cholangitis (PSC) , acetaminophen toxicity, hepatotoxicity) , kidney damage/injury (nephritis, renal transplant, surgery, administration of nephrotoxic drugs e.g. cisplatin, acute kidney injury (AKI) ) , Celiac disease, autoimmune idiopathic thrombocytopenic purpura, transplant rejection, ischemia reperfusion injury of solid organs, sepsis, systemic inflammatory response syndrome (SIRS) , cerebrovascular accident (CVA, stroke) , myocardial infarction (MI) , atherosclerosis, Huntington's disease, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS) , spinal muscular atropy (SMA) , allergic diseases (including asthma and atopic dermatitis) , multiple sclerosis, type I diabetes, Wegener's granulomatosis, pulmonary sarcoidosis, Behcet's disease, interleukin-1 converting enzyme (ICE, also known as caspase-1) associated fever syndrome, chronic obstructive pulmonary disease (COPD) , tumor necrosis factor receptor-associated periodic syndrome (TRAPS) , periodontitis, linear ubiquitin chain assembly complex (LUBAC) deficiency syndrome, hematological and solid organ malignancies, bacterial infections and viral infections (such as tuberculosis and influenza) , and Lysosomal storage diseases.

DETAILED DESCRIPTION

I. Definitions

A dash ( "-" ) at the left hand side of a substituent is used to indicate a point of attachment for a substituent. For example, -CH ₂-phenyl is attached through the carbon atom of CH ₂.

The term "alkyl" herein refers to a straight or branched hydrocarbon chain. The symbol of C subscripted with a number range that precedes the term “alkyl” stands for the number of carbons in the alkyl. For example, C _1-5 alkyl represents an alkyl containing 1, 2, 3, 4, or 5 carbon atoms. Examples of C _1-5 alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, and pentyl.

The term "cycloalkyl" refers to a saturated and partially unsaturated cyclic hydrocarbon group only having carbon atom as the ring atom. The symbol of C subscripted with a number range that precedes the term “cycloalkyl” stands for the carbon ring numbers in the cycloalkyl. For example, C _3-5 cycloalkyl represents a cycloalkyl containing 3, 4, or 5 carbon ring atoms, i.e., cyclopropyl, cyclobutyl, or cyclopentyl. The ring may be saturated or have one or more double bonds (i.e. partially unsaturated) , but not fully conjugated.

The term “haloalkyl” refers to an alkyl substituted by 1, 2, 3, 4, or 5 halogens, wherein the halogen is Cl or F. “HaloC _1-3alkyl” refers to C _1-3 alkyl substituted by 1, 2, 3, 4, or 5 Cl or F. Exemplary haloC _1-3 alkyl includes -CF ₃, -CHF ₂, -CH ₂CF ₃, etc.

The term "heteroaryl" refers to 5-to 8-membered aromatic, monocyclic rings containing one or more, for example, from 1 to 4, or, in some embodiments, from 1 to 3, heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon; 8-to 12-membered fused bicyclic rings containing one or more, for example, from 1 to 4, or, in some embodiments, from 1 to 3, heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon and wherein at least one heteroatom is present in an aromatic ring.

Exemplary heteroaryl include, but are not limited to, pyridinyl, pyrazinyl, pyrazinyl, pyrimidinyl, pyrazolyl, imidazolinyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, tetrazolyl, thienyl, benzothienyl, furyl, benzofuryl, benzoimidazolinyl, indolinyl, pyridizinyl, triazolyl, quinolinyl, pyrazolyl, and 5, 6, 7, 8-tetrahydroisoquinoline.

The term “heterocyclyl" means a single, fused, spiro, or bridged bicyclic or multicyclic, ring, with 5 to 12 ring atoms, containing at least 2 carbon atoms in addition to 1-3 heteroatoms independently selected from oxygen, sulfur, and nitrogen. When there is more than one ring system, at least one of the rings is saturated or not fully conjugated, contains at least one heteroatom, and is the point of attachment ring. The point of the attachment may be carbon or heteroatom in the heterocyclic ring.

Exemplary heterocyclyl includes but are not limited to pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, tetrahydro-furanyl, 5, 6, 7, 8-tetrahydroimidazo [1, 2-a] pyrazinyl, tetrahydro-2H-pyranyl, 8-oxa-3-azabicyclo [3.2.1] octanyl, 3-oxa-9-azaspiro [5.5] undecanyl, 7-oxa-2-azaspiro [3.5] nonanyl, and 2-oxa-7-azaspiro [3.5] nonanyl.

‘Oxo” means = (O) group.

“Pharmaceutically acceptable salt” refers to a salt form of a compound (e.g., a drug) having at least one group capable for salt formation that causes no significant adverse toxicological effects to the subject. Pharmaceutically acceptable salts include, for example, salts prepared by reaction with an inorganic acid, organic acid, or a base depending on the nature of the compound (e.g., drug) . The inorganic acid can be hydrochloric acid, hydrobromic acid, carbonic acid, sulfuric acid, phosphoric acid, and the like; the organic acid can be fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, methanesulfonic acid and the like. The base that can form a salt with an acid drug can be an amine containing compound or inorganic base such as sodium hydroxide, sodium carbonate, and the like. Suitable pharmaceutically acceptable salt forms can e found in, for example, Handbook of Pharmaceutical Salts: Properties, Selection and Use, Weinheim/Zürich: Wiley-VCH/VHCA, 2002; P.H. Stahl and C.G. Wermuth, Eds.

As used herein, a “therapeutically effective amount” refers to the amount that, when administered to a subject for treatment of a disease, is sufficient to cause a desired treatment effect in the subject, including for example, alleviation of the symptoms or stop of the progression of the disease.

The terms “treating” , “treatment” , or “treat” (of) a disease refers to slowing or arresting the development of a disease, providing relief from the symptoms or side-effects of the disease, and/or causing regression of the disease. The terms also refers to reduction of the occurrence of the disease in the subject when compared with a subject without the treatment.

A "pharmaceutically acceptable excipient" means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable, and includes a carrier or an excipient that is acceptable for veterinary use as well as human pharmaceutical use. "Apharmaceutically acceptable excipient" as used in the specification and claims includes both one and more than one such excipient.

The term “subject” as used herein refers to animal (such as mammal) or human. Compound of formula I and a pharmaceutically acceptable salt thereof as described herein include, but are not limited to, their solvates, optical isomers, racemates, and other mixtures thereof. In those situations, the single enantiomers or diastereomers, i.e., optically active forms, can be obtained by asymmetric synthesis or by resolution of the racemates or mixtures of diastereomers. Resolution of the racemates or mixtures of diastereomers can be accomplished, for example, by conventional methods such as crystallization in the presence of a resolving agent, or chromatography, using, for example a chiral high-pressure liquid chromatography (HPLC) column. Where compounds described herein exist in various tautomeric forms, the term "compound" is intended to include all tautomeric forms of the compound.

II. Compounds and Uses Thereof

Disclosed is a compound of formula I or a pharmaceutically acceptable salt thereof:

wherein

R ₁ is H, F, CN, CF ₃, or Cl,

R ₂ is H, -CH ₃, -CD ₃, or cyclopropyl,

A ring is a bicyclic fused ring system represented by

in which N ring is fused to the amide-containing seven-membered ring,

M is a 5 or 6 membered heteroaryl or phenyl each of which is optionally substituted by one or two groups selected from oxo, Cl, -O-C _1-3 alkyl, OH, CN, -C _1-3 alkyl, -S-C _1-3 alkyl, phenyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b,

R _a is H, CH ₃, or CH ₂CH ₃,

X is O or CH ₂,

B is

in which the amide nitrogen is attached to the amide-containing seven membered ring, X ₁ is CH or N, R _c is selected from H, -CH ₃, -CD ₃, -CH ₂CF ₃, -CH=CH ₂, and cyclopropyl, and R _c is attached to one of the nitrogen of the 5-membered ring,

L is C=O, CHCH ₃, C (CH ₃) ₂, O, CHOH, CH ₂ or

and

provided that when B is

In some embodiments, M ring is an oxazoline, pyrimidine, pyridine, or pyrazine, each of which is optionally substituted by one or two groups selected from oxo, C _1-3 alkyl, -S-C _1-3 alkyl, 5 to 12 membered heterocycyl optionally substituted with one or more W, and -NR _aR _b, wherein W and R _a and R _b are as disclosed herein.

In some embodiments, the 5 to 12 membered heterocycyl optionally substituted with one or two W is selected from pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, 5, 6, 7, 8-tetrahydroimidazo [1, 2-a] pyrazinyl, tetrahydro-2H-pyranyl, 8-oxa-3-azabicyclo [3.2.1] octanyl, 3-oxa-9-azaspiro [5.5] undecanyl, 7-oxa-2-azaspiro [3.5] nonanyl, and 2-oxa-7-azaspiro [3.5] nonanyl, each of which is optionally substituted with one or two W as disclosed herein.

In some embodiments, when the 5 to 12 membered heterocycyl optionally substituted with one or two W is a nitrogen-containing 5 to 12 membered heterocycyl, the 5 to 12 membered heterocycyl is attached to M ring through the nitrogen.

In some embodiments, the optionally substituted 5 or 6 membered heterocyclyl for R _b is optionally substituted pyrrolidinyl, piperazinyl, morpholinyl or piperidinyl.

In some embodiments, B is

In some embodiments, L is CH ₂ or

In some embodiments, A ring is

wherein R ₃ is selected from H, -S-C _1-3 alkyl, phenyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, R ₄ is H, F, or Cl, and wherein the 5 to 12 membered heterocycyl, W, R _a and R _b are as disclosed herein.

In some embodiments, A ring is

wherein R ₅ is selected from H, OH, Cl, -O-C _1-3 alkyl, -S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, R ₄ is H, F, or Cl, and wherein the 5 to 12 membered heterocycyl, W, R _a and R _b are as disclosed herein.

In some embodiments, R ₅ is selected from C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, and wherein the 5 to 12 membered heterocycyl, W, R _a and R _b are as disclosed herein.

In some embodiments, R ₅ is selected from 5 or 6 membered heterocyclyl and -NH-C _3-5 cycloalkyl. In some embodiments, R ₅ is selected from -NH-cyclopropyl and morpholinyl.

In some embodiments, A ring is

R ₄ is H, F, or Cl, R ₆ is selected from H, CN, -S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, R ₄ is H, F, or Cl, and wherein the 5 to 12 membered heterocycyl, W, R _a and R _b are as disclosed herein.

In some embodiments, R ₆ is selected from H, , CN, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, and wherein the 5 to 12 membered heterocycyl, W, R _a and R _b are as disclosed herein.

In some embodiments, R ₆ is selected from H, CN, 5 or 6 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, W, R _a and R _b are as disclosed herein.

In some embodiments, R ₆ is selected from H, CN, morpholinyl, piperidinyl, pyrrolidinyl, -NH-cyclopropyl, -NH (CH ₂) _2-3OH, -NH (CH ₂) _2-3OCH ₃, piperazinyl, -NHCH ₂-cyclopropyl, wherein the piperidinyl, pyrrolidinyl, and piperazinyl, are optionally substituted with -C _3-5 cycloalkyl or -C _1-3 alkyl.

In some embodiments, A ring is

Z is CH or N, R ₇ and R ₈ are independently H, F, or Cl, provided that at least one of R ₇ and R ₈ is not H, and R ₉ is selected from H, -S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, and wherein the 5 to 12 membered heterocycyl, W, R _a and R _b are as disclosed herein.

In some embodiments, R ₉ is selected from C _3-6 cycloalkyl optionally substituted with one or two W, 5 or 6 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, and wherein the 5 to 12 membered heterocycyl, W, R _a and R _b are as disclosed herein.

In some embodiments, R ₉ is selected from morpholinyl, piperidinyl, pyrrolidinyl, -NH-cyclopropyl, -NH-tetrahydro-2H-pyranyl, -NH (CH ₂) _2-3OH, - NH (CH ₂) _2-3OCH ₃, piperazinyl, -NHCH ₂-cyclopropyl, wherein the piperidinyl, pyrrolidinyl, and piperazinyl, are optionally substituted with -C _3-5 cycloalkyl or -C _1-3 alkyl.

In some embodiments, R ₉ is H or -C _1-3 alkyl.

In some embodiments, A ring is

wherein R ₁₀ is selected from H, -C _1-3 alkyl, -S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, and wherein the 5 to 12 membered heterocycyl, W, R _a and R _b are as disclosed herein.

In some embodiments, R ₁₀ is H or C _3-5 cycloalkyl.

In some embodiments, A ring is

R ₁₁ is selected from H, -C _1-3 alkyl, -S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, and wherein the 5 to 12 membered heterocycyl, W, R _a and R _b are as disclosed herein.

In some embodiments, R ₁₁ is selected from 5 to 6 membered heterocycyl optionally substituted with one or two W and -NR _aR _b, W, R _a and R _b are as disclosed herein.

In some embodiment, R ₁₁ is morpholinyl, piperazinyl, -NH-cyclopropyl, or -NH-tetrahydro-2H-pyranyl, wherein piperazinyl is optionally substituted with C _3-5 cycloalkyl.

In some embodiments, A ring is

R ₁₂ is selected from H, CN, -C _1-3 alkyl, -S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, and wherein the 5 to 12 membered heterocycyl, W, R _a and R _b are as disclosed herein.

In some embodiments, R ₁₂ is selected from -NR _aR _b and 5 to 6 membered heterocycyl optionally substituted with one or two W, W, R _a and R _b are as disclosed herein.

In some embodiments, R ₁₂ is -NH (CH ₂) _2-3OH, -NH (CH ₂) _2-3-OCH ₃, morpholinyl, -NHCH ₂-cyclopropyl, piperazinyl, -NH-cyclopropyl, or -NH-tetrahydro-2H-pyranyl, wherein piperazinyl is optionally substituted with C _1-3 alkyl or C _3-5 cycloalkyl.

In some embodiments, A ring is

Z is CH or N.

In some embodiments, A ring is

R ₁₃ is C _1-3 alkyl or C _3-5 cycloalkyl.

In some embodiments, A ring is selected from

Also provided is a compound selected from

or a pharmaceutically acceptable salt thereof.

Also provided is a compound selected from

or a pharmaceutically acceptable salt thereof.

Also provided is a compound selected from

a pharmaceutically acceptable salt thereof.

The compound disclosed herein may have improved stability and pharmacokinetic profiles over existing RIP1 kinase inhibitors.

Compounds disclosed herein (the term “compound (s) disclosed herein” includes pharmaceutically acceptable salt thereof) will be administered in a therapeutically effective amount by any of the accepted administration modes for agents that serve similar utilities. Therapeutically effective amount of the compounds disclosed herein may range from 0.01 to 500 mg per kg subject body weight, which can be administered in single or multiple doses per day. For oral administration, the compositions can be provided in the form of tablets or capsules containing 1.0 to 1000 mg of the compound disclosed herein, such as, 1.0, 5.0, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, and 1000 mg of the compound disclosed herein.

In addition to oral administration, the compound disclosed herein can also be administered as pharmaceutical compositions by, for example, transdermal, intranasal, suppository, intramuscular, intravenous or subcutaneous administration. The pharmaceutical composition comprises the compound disclosed herein and a pharmaceutically acceptable excipient. When prepared for unit dosage form, the pharmaceutical compositions can comprise from 1 mg to 1000 mg of the compound disclosed herein.

Exemplary solid pharmaceutical excipient includes starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, dried skim milk and the like. Liquid and semisolid excipients may be selected from glycerol, propylene glycol, water, ethanol and various oils, including those of petroleum, animal, vegetable or synthetic origin, e g, peanut oil, soybean oil, mineral oil, sesame oil, etc. Preferred liquid carriers, particularly for injectable solutions, include water, saline, aqueous dextrose, and glycols.

Other suitable pharmaceutical excipients and their formulations are described in Remington's Pharmaceutical Sciences, edited by E.W. Martin (Mack Publishing Company, 20th ed., 2000) .

The compounds discloses herein can be used to achieve therapeutic effects in a subject with disease or disorder mediated by RIP1 kinase, which include cancer and inflammatory diseases. For example, the disease or disorder can be selected from inflammatory bowel diseases, Crohn's disease, ulcerative colitis, psoriasis, retinal detachment, retinitis pigmentosa, macular degeneration, pancreatitis, atopic dermatitis, arthritis (including rheumatoid arthritis, osteoarthritis, spondylarthritis, gout, systemic onset juvenile idiopathic arthritis (SoJIA) , psoriatic arthritis) , systemic lupus erythematosus (SLE) , Sjogren's syndrome, systemic scleroderma, anti-phospholipid syndrome (APS) , vasculitis, liver damage/diseases (non-alcohol steatohepatitis, alcohol steatohepatitis, autoimmune hepatitis autoimmune hepatobiliary diseases, primary sclerosing cholangitis (PSC) , acetaminophen toxicity, hepatotoxicity) , kidney damage/injury (nephritis, renal transplant, surgery, administration of nephrotoxic drugs e.g. cisplatin, acute kidney injury (AKI) ) , Celiac disease, autoimmune idiopathic thrombocytopenic purpura, transplant rejection, ischemia reperfusion injury of solid organs, sepsis, systemic inflammatory response syndrome (SIRS) , cerebrovascular accident (CVA, stroke) , myocardial infarction (MI) , atherosclerosis, Huntington's disease, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS) , spinal muscular atropy (SMA) , allergic diseases (including asthma and atopic dermatitis) , multiple sclerosis, type I diabetes, Wegener's granulomatosis, pulmonary sarcoidosis, Behcet's disease, interleukin-1 converting enzyme (ICE, also known as caspase-1) associated fever syndrome, chronic obstructive pulmonary disease (COPD) , tumor necrosis factor receptor-associated periodic syndrome (TRAPS) , periodontitis, linear ubiquitin chain assembly complex (LUBAC) deficiency syndrome, hematological and solid organ malignancies, bacterial infections and viral infections (such as tuberculosis and influenza) , and Lysosomal storage diseases.

In some embodiments, the disease or disorder is selected from inflammatory bowel disease, Crohn's disease, ulcerative colitis, psoriasis, retinal detachment, retinitis pigmentosa, macular degeneration, pancreatitis, atopic dermatitis, rheumatoid arthritis, spondyloarthritis, gout, SoJIA, systemic lupus erythematosus, Sjogren's syndrome, systemic scleroderma, anti-phospholipid syndrome, vasculitis, osteoarthritis, non-alcohol steatohepatitis, alcohol steatohepatitis, autoimmune hepatitis autoimmune hepatobiliary diseases, primary sclerosing cholangitis, nephritis, Celiac disease, autoimmune ITP, transplant rejection, ischemia reperfusion injury of solid organs, sepsis, systemic inflammatory response syndrome, cerebrovascular accident, myocardial infarction, Huntington's disease, Alzheimer's disease, Parkinson's disease, allergic diseases, asthma, atopic dermatitis, multiple sclerosis, type I diabetes, Wegener's granulomatosis, pulmonary sarcoidosis, Behcet's disease, interleukin-1 converting enzyme associated fever syndrome, chronic obstructive pulmonary disease, tumor necrosis factor receptor-associated periodic syndrome, and periodontitis.

In some embodiments, the disease or disorder is selected from inflammatory bowel disease, Crohn's disease, ulcerative colitis, multiple sclerosis, arthritis, rheumatoid arthritis, osteoarthritis, and psoriasis

The compound disclosed herein may be administered in combination with other known anti-inflammatory agents for any indications above. These anti-inflammatory agents can be small molecules or antibodies. Exemplary anti-inflammatory agents include corticosteroids, 5-aminosalicyclic acid, tofacitinib, rituximab, adalimumab, etc.

EXAMPLES

The following examples are illustrative in nature and are in no way intended to be limiting.

Example 1

Synthesis of the Compounds

Scheme I below describes an illustrative synthetic scheme for preparation of compound 20 disclosed herein.

Scheme I: Synthesis scheme for compound 20:

General procedure for preparation of compound A-2

To a solution of compound A-1 (12.0 g, 69.7 mmol, 1.00 eq) , DIEA (18.0 g, 139 mmol, 24.3 mL, 2.00 eq) and tert-butyl N-amino carbamate (9.67 g, 73.2 mmol, 1.05 eq) in DMF (100 mL) were added EDCI (14.0 g, 73.2 mmol, 1.05 eq) and 1-oxidopyridin-1-ium-2-ol (8.13 g, 73.2 mmol, 1.05 eq) . The solution was stirred at 70 ℃ for 4 h. The residue was poured into water (500 mL) . The aqueous phase was extracted with ethyl acetate (500 mL*3) . The combined organic phase was washed with brine (500 mL*2) , dried with anhydrous Na ₂SO ₄, filtered and concentrated in vacuum to afford compound A-2 (20.0 g, crude) as a yellow oil which was used for next step directly without further purification.

General procedure for preparation of compound A-3

To a solution of compound A-2 (20.0 g, 55.8 mmol, 1.00 eq) in dioxane (100 mL) was added HCl/dioxane (4 M, 200 mL, 14.3 eq) at 0 ℃. The solution was stirred at 20 ℃ for 4 h. TLC showed starting material was consumed completely. The reaction mixture was concentrated in reduced pressure at 50 ℃ to afford compound A-3 (15.0 g, crude, HCl salt) as a yellow solid which was used for next step directly without further purification.

General procedure for preparation of compound A-4

To a solution of compound A-3 (1.00 g, 4.30 mmol, 1.00 eq) and TEA (869 mg, 8.59 mmol, 1.20 mL, 2.00 eq) in EtOH (10 mL) was added ethyl 2-ethoxy-2-imino-acetate (623 mg, 4.30 mmol, 1.00 eq) . The reaction mixture was stirred at 50 ℃ for 2 h. Solid precipitated out. The reaction mixture was filtered to afford desired product. The crude product was used for next step directly without purification. Compound A-4 (1.00 g, crude) was obtained as a white solid.

LC-MS: 286.0

General procedure for preparation of compound A-5

A solution of compound A-4 (1.00 g, 3.16 mmol, 1.00 eq) in EtOH (20 mL) was stirred at 70 ℃ for 4 h. The reaction mixture was concentrated in reduced pressure at 20 ℃. Solid was filtered out and the filtered cake was discarded. The filtrate was concentrated in reduced pressure to afford the crude product Compound A-5 (0.50 g, 1.68 mmol, 53.3%yield, 90.0%purity) as a yellow oil which was used for next step directly without purification.

LC-MS: 267.9

General procedure for preparation of A-6

To a solution of compound A-5 (0.10 g, 374 umol, 1.00 eq) in THF (1 mL) and H ₂O (1 mL) was added LiOH (2 M, 374 uL, 2.00 eq) . The reaction was stirred at 20 ℃ for 4 h. The solution was adjusted by HCl (1 N) to 7 -8. The reaction mixture was concentrated in reduced pressure at 40 ℃ to afford the crude product compound A-6 (100 mg, crude) as a yellow solid which was used for next step directly without purification.

¹H NMR: (400 MHz, DMSO-d ₆) δ ppm 7.44 -7.63 (m, 6 H)

LC-MS: 240.1

General procedure for preparation of A-7

To a solution of compound A-6 (0.20 g, 673 umol, 1.00 eq) in H ₂O (2 mL) and THF (2 mL) was added LiOH (64.5 mg, 2.69 mmol, 4.00 eq) . The reaction was stirred at 50 ℃ for 16 h. The solution was concentrated in reduced pressure at 40 ℃. The crude product was used for next step directly without purification. A-7 (0.20 g, crude) was obtained as a yellow solid.

LC-MS: 218.1

General procedure for preparation of compound 20

To a solution of A-8 (100 mg, 429 umol, 1.00 eq) , A-7 (119 mg, 428 umol, 1.00 eq) and DIEA (111 mg, 857 umol, 149 uL, 2.00 eq) in DMF (3 mL) were added 1-oxidopyridin-1-ium-2-ol (50.0 mg, 450.21 umol, 1.05 eq) and EDCI (86.3 mg, 450 umol, 1.05 eq) . The reaction mixture was stirred at 30 ℃ for 16 h before it was poured into water (5 mL) . The aqueous phase was extracted with ethyl acetate (5 mL*3) . The combined organic phase was discarded. The aqueous phase was concentrated in vacuum. The crude product was purified by prep-HPLC (column: Phenomenex Gemini-NX 150*30mm*5um; mobile phase: [water (0.05%NH ₃H ₂O + 10 mM NH ₄HCO ₃) -ACN] ; B%: 10%-30%, 8 min) to give compound 20 (15.5 mg, 34.6 umol, 8.08%yield, 96.7%purity) as a white solid. ¹H NMR: (400 MHz, DMSO-d6) δ ppm 8.53 (br d, J=7.78 Hz, 2 H) , 8.16 -8.30 (m, 2 H) , 7.66 -7.77 (m, 2 H) , 7.45 -7.61 (m, 3 H) , 5.05 -5.25 (m, 1 H) , 4.69 -4.91 (m, 1 H) , 4.40 (br t, J=10.54 Hz, 1 H) , 3.56 (s, 3 H)

LC-MS: 433.1

Scheme II below describes an illustrative synthetic scheme for preparation of compound 21 disclosed herein.

Scheme II: Synthesis scheme for compound 21:

General procedure for preparation of compound A-10:

To a solution of compound A-9 (10 g, 61.6 mmol, 1 eq. ) and tert-butyl N-amino carbamate (8.15 g, 61.6 mmol, 1 eq. ) in DMF (100 mL) was added 1-oxidopyridin-1-ium-2-ol (7.19 g, 64.7 mmol, 1.05 eq) , EDCI (12.4 g, 64.7 mmol, 1.05 eq) and DIPEA (15.9 g, 123 mmol, 21.5 mL, 2 eq. ) . The mixture was stirred at 60 ℃ for 2 h. TLC (petroleum ether/EtOAc=1/1, I ₂) showed the acid was consumed completely. The mixture was diluted with EtOAc (200 mL) , washed with H ₂O (200 mL) , sat. NaHCO ₃ (50 mL) , brine (50 mL*3) , dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give a light yellow gum. The gum solidified by standing at r.t overnight. The crude product compound A-10 (16.5 g, 59.7 mmol, 96.8%yield) was obtained as a light yellow solid and was directly used in the next step without further purification.

¹H NMR: ES12645-155-P1 (400 MHz, DMSO-d6) δ ppm 8.86 (br s, 1 H) , 8.61 (br s, 1 H) , 7.19 -7.37 (m, 5 H) , 1.37 (s, 9 H) , 1.33 (m, 2 H) , 0.99 -1.05 (m, 2 H)

General procedure for preparation of compound A-11

To a solution of compound A-10 (16 g, 57.9 mmol, 1 eq. ) in EtOAc (150 mL) was added HCl/EtOAc (4 M, 150 mL, 10.3 eq) at 0 ℃. The mixture was then stirred at 25 ℃ for 0.5 h. Large amount of white solid precipitated. TLC (petroleum ether/EtOAc=1/1, UV) showed the starting material was consumed completely. The mixture was filtered and the filter cake was rinsed with EtOAc (100 mL) and then dried under vacuum to give compound A-11 (10.2 g, 47.9 mmol, 82.8%yield, HCl salt) as a white solid, which was directly used in the next step without further purification.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 10.29 (br s, 1 H) , 10.02 (s, 1 H) , 7.28 -7.41 (m, 5 H) , 1.42 -1.47 (m, 2 H) , 1.12 -1.17 (m, 2 H)

General procedure for preparation of compound A-12

To a solution of compound A-11 (8 g, 37.6 mmol, 1 eq, HCl salt) in EtOH (70 mL) and THF (100 mL) was added ethyl 2-ethoxy-2-imino-acetate (6.01 g, 41.3 mmol, 1.1 eq. ) and TEA (11.4 g, 113 mmol, 15.7 mL, 3 eq. ) . The mixture was stirred at 25 ℃ under N ₂ for 6 h. The mixture was filtered and the filter cake was rinsed with DCM (30 mL) and then dried under vacuum to give compound A-12 (9.4 g, 34.1 mmol, 90.8%yield) as a white solid, which was directly used in the next step without further purification.

General procedure for preparation of compound A-13

A suspension of A-12 (10 g, 36.3 mmol, 1 eq. ) in xylene (120 mL) was heated at 160 ℃ and azeotropic distillation to remove water by Dean-Stark trap for 6 h. The mixture was concentrated to give a yellow residue. The residue was dissolved into DCM (100 mL) , washed with brine (50 mL) , dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give a crude product. The crude product was purified by flash silica gel chromatography (

80 g

Silica Flash Column, Eluent of 0～3%MeOH/DCM gradient @60 mL/min) to give compound A-13 (3.8 g, 14.7 mmol, 40.6%yield) as a white solid.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 14.17 (br s, 1 H) , 7.29 -7.36 (m, 5 H) , 4.23 -4.34 (m, 2 H) , 1.42 -1.53 (m, 2 H) , 1.33 -1.39 (m, 2 H) , 1.28 (t, J=7.09 Hz, 3 H)

General procedure for preparation of compound A-14:

To a suspension of compound A-13 (4 g, 15.5 mmol, 1 eq. ) in THF (50 mL) was added a solution of LiOH. H ₂O (2.61 g, 62.2 mmol, 4 eq. ) in H ₂O (50 mL) . The mixture was stirred at 25 ℃ for 3 h. LC-MS showed the reaction was completed. The mixture was concentrated to remove THF. The aqueous layer was extracted with EtOAc (30 mL*2) . The combined organic layer was discarded, and the aqueous layer was cooled to 0 ℃, acidified with 2 N HCl to pH～4-5. Solid precipitated. The white solid was collected by filtration, and then rinsed with water (30 mL) , dried under vacuum to give A-14 (2.8 g, 11.6 mmol, 74.8%yield, 95.2%purity) as a white solid.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 14.05 (br s, 1 H) 7.24 -7.34 (m, 5 H) 1.47 (m, 2 H) 1.32 (m, 1 H) 2 H) , 1.28 (t, J=7.09 Hz, 3 H)

LC-MS: 230.2

General procedure for preparation of compound 21

To a mixture of A-8 (70 mg, 300 umol, 1 eq. ) , A-14 (68.8 mg, 300 umol, 1 eq. ) in DMF (3 mL) was added EDCI (69.0 mg, 360 umol, 1.2 eq. ) and 1-oxidopyridin-1-ium-2-ol (40.0 mg, 360 umol, 1.2 eq. ) and TEA (91.1 mg, 900 umol, 125 uL, 3 eq. ) . The mixture was stirred at 25 ℃ for 18 h. The mixture turned yellow clear. LC-MS showed the reaction was completed. The mixture was diluted with EtOAc (30 mL) , washed with H ₂O (30 mL) , brine (30 mL*3) . The separated organic layer was dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give yellow residue. The residue was purified by prep-HPLC (column: Phenomenex Gemini-NX 150*30mm*5um; mobile phase: [water (0.05%NH _3. H ₂O+10mM NH ₄HCO ₃) -ACN] ; B%: 25%-45%, 8min) to give compound 22 (12.6 mg, 27.6 umol, 9.20%yield, 97.4%purity) as a white solid.

LCMS: 445.2

¹H NMR (400 MHz, CDCl ₃) : δ ppm 11.44 (s, 1 H) , 8.18 (s, 1 H) , 8.10 (br d, J=7.25 Hz, 1 H) , 7.69 (s, 1 H) , 7.48 (s, 1 H) , 7.30 (s, 1 H) , 7.25 -7.28 (m, 2 H) , 7.21 (br s, 1 H) , 5.00 -5.12 (m, 1 H) , 4.72 -4.83 (m, 1 H) , 4.29 -4.38 (m, 1 H) , 4.15 (s, 2 H) , 3.53 (s, 3 H)

Scheme III below describes an illustrative synthetic scheme for preparation of compound 23 disclosed herein.

Scheme III: Synthesis scheme for compound 23:

General procedure for preparation of compound A-16

To a solution of compound A-15 (10.6 g, 31.7 mmol, 1 eq) in DCM (200 mL) was added dropwise TFA (50 mL) at 25 ℃. The mixture was stirred at 25 ℃ for 1 h. The mixture was concentrated to give compound A-15B (11.0 g, 31.7 mmol, TFA salt) as a brown gum, which was directly used in the next step without further purification.

To a solution of compound A-15B (11.0 g, 31.5 mmol, 1 eq, TFA salt) in DCM (300 mL) was added TEA (9.59 g, 94.7 mmol, 13.2 mL, 3 eq) . Then TFAA (7.96 g, 37.9 mmol, 5.27 mL, 1.2 eq) was added under N ₂ at 0 ℃. After the addition. The mixture was stirred at 0 ℃ for 20 min. The mixture was washed with brine (100 mL*2) , dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give compound A-16 (9.64 g, 29.2 mmol, 92.4%yield) as a yellow solid, which was directly used in the next step without further purification.

¹H NMR: (400 MHz, CDCl ₃) δ ppm 7.93 (s, 1 H) , 7.54 (br d, J=5.77 Hz, 1 H) , 6.95 (d, J=8.28 Hz, 1 H) , 6.65 -6.81 (m, 2 H) , 6.04 (ddt, J=17.29, 10.57, 5.27, 5.27 Hz, 1 H) , 5.43 (dd, J=17.19, 1.38 Hz, 1 H) , 5.32 (dd, J=10.54, 1.25 Hz, 1 H) , 4.88 (dt, J=10.54, 6.40 Hz, 1 H) , 4.76 (dd, J=10.16, 6.65 Hz, 1 H) , 4.53 (d, J=5.27 Hz, 2 H)

General procedure for preparation of compound A-17

A solution of compound A-16 (8.20 g, 24.8 mmol, 1 eq) in TFAA (150 mL) was cooled to -30 ℃ under N ₂. Then HNO ₃ (2.53 g, 27.3 mmol, 1.81 mL, 68%purity, 1.1 eq) was added dropwise while keeping the internal temperature around -30 ℃. After the addition, the mixture was stirred at -30 ℃ for 30 min. The mixture was poured onto ice water (1000 mL) , solid precipitated. The solid was collected by filtration, dried under vacuum to give compound A-17 (9.1 g, crude) as a brown solid, which was directly used in the next step without further purification. ¹H NMR: (400 MHz, CDCl ₃) δ ppm 9.01 (s, 1 H) , 7.76 (s, 1 H) , 7.53 (br d, J=6.02 Hz, 1 H) , 6.84 (s, 1 H) , 5.97 -6.18 (m, 1 H) , 5.52 (br d, J=17.32 Hz, 1 H) , 5.38 (br d, J=10.54 Hz, 1 H) , 4.94 (dt, J=10.35, 5.49 Hz, 1 H) , 4.74 (dd, J=10.54, 5.02 Hz, 1 H) , 4.68 (br d, J=5.02 Hz, 2 H) , 4.37 -4.43 (m, 1 H) .

General procedure for preparation of compound A-18:

To a solution of compound A-17 (8.65 g, 18.4 mmol, 1 eq) in DCM (200 mL) was added Pd (PPh ₃) ₄ (8.52 g, 7.38 mmol, 0.4 eq) , Bu ₃SnH (16.1 g, 55.3 mmol, 14.6 mL, 3 eq) , AcOH (6.64 g, 110.64 mmol, 6.33 mL, 6 eq) under N ₂. The mixture was stirred at 25 ℃ for 1 h. The mixture was concentrated and the residue was triturated in petroleum ether (500 mL) , filtered and the filtrate was discarded. The filter cake was dried to give a black residue. The residue was purified by flash silica gel chromatography (

330 g

Silica Flash Column, Eluent of 0～3%MeOH/DCM gradient @100 mL/min) to give a yellow solid. ¹H NMR showed Bu ₃SnH residual. To this solid was added t-BuOMe (50 mL) , stirred for 10 minutes, filtered and the filtrate was concentrated to give compound A-18 (3.55 g, 10.6 mmol, 57.4%yield) as a yellow solid.

¹H NMR: (400 MHz, CDCl ₃) δ ppm10.98 (br s, 1 H) , 10.24 (s, 1 H) , 9.84 (d, J=7.78 Hz, 1 H) , 7.79 (s, 1 H) , 6.79 (s, 1 H) , 4.78 (td, J=8.53, 5.52 Hz, 1 H) , 4.43 -4.58 (m, 2 H) .

General procedure for preparation of compound A-19

To a solution of compound A-18 (1.46 g, 4.35 mmol, 1 eq) in MeOH (40 mL) was added Pd/C (200 mg, 10%purity) under N ₂. The mixture was degassed and purged with H ₂ for 3 times. Then the mixture was stirred at 25 ℃ under H ₂ (15 psi) for 2 h. The mixture was filtered through a pad of Celite and the filtrate was concentrated to give compound A-19 (1.22 g, 4.00 mmol, 91.8%yield) as a yellow solid, which was directly used in the next step without further purification.

General procedure for preparation of compound A20

A solution of compound A-19 (1.21 g, 3.96 mmol, 1 eq) in trimethoxymethane (19.3 g, 182 mmol, 20 mL, 46.0 eq) was stirred at 130 ℃ for 4 h. The mixture was concentrated to give a yellow residue which was purified by flash silica gel chromatography (

20 g

Silica Flash Column, Eluent of 0～20%THF/DCM gradient @35 mL/min) to give compound A-20 (0.88 g, 2.79 mmol, 70.4%yield) as a yellow solid.

¹H NMR: (400 MHz, DMSO-d ₆) δ ppm 10.25 (s, 1 H) , 9.78 (d, J=7.95 Hz, 1 H) , 8.76 (s, 1 H) , 7.68 (s, 1 H) , 7.51 (s, 1 H) , 4.75 (dt, J=10.73, 8.02 Hz, 1 H) , 4.45 -4.63 (m, 2 H)

General procedure for preparation of compound A-8

To a solution of A-20 (70 mg, 222 umol, 1 eq) in THF (3 mL) and H ₂O (3 mL) was added K ₂CO ₃ (46.0 mg, 333 umol, 1.5 eq) . The mixture was stirred at 30 ℃ for 18 h. The mixture was concentrated to remove THF. The remaining aqueous layer was purified by prep-HPLC (column: Phenomenex Gemini-NX 150*30mm*5um; mobile phase: [water (0.05%NH ₃H ₂O+10mM NH ₄HCO ₃) -ACN] ; B%:20%-40%, 8 min) to give A-8 (6.3 mg, 28.7 umol, 12.9%yield, 100%purity) as a yellow solid.

LC-MS: 220.0

¹H NMR: (400 MHz, DMSO-d6) δ ppm 9.91 (br s, 1 H) , 8.72 (s, 1 H) , 7.60 (s, 1 H) , 7.42 (s, 1 H) , 4.35 (dd, J=10.21, 7.40 Hz, 1 H) , 4.05 (t, J=10.82 Hz, 1 H) , 3.62 (dd, J=11.37, 7.34 Hz, 1 H) , 2.01 (br s, 2 H)

General procedure for preparation of compound 23

To a mixture of A-8 (50 mg, 228 umol, 1 eq) , 5-benzyl-4H-1, 2, 4-triazole-3-carboxylic acid (55.6 mg, 273 umol, 1.2 eq) in DMF (2 mL) was added EDCI (52.5 mg, 274 umol, 1.2 eq) and 1-oxidopyridin-1-ium-2-ol (30.4 mg, 274 umol, 1.2 eq) and TEA (69.2 mg, 684 umol, 95.2 uL, 3 eq) . The mixture was stirred at 50 ℃ for 1 h and then was concentrated to remove DMF. The yellow residue was partitioned between EtOAc (100 mL) and water (50 mL) . The aqueous layer was extracted with EtOAc (50 mL*4) . The combined organic layer was dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give a yellow residue. The residue was purified by prep-HPLC (column: Phenomenex Gemini-NX 150*30mm*5um; mobile phase: [water (0.05%NH ₃H ₂O+10mM NH ₄HCO ₃) -ACN] ; B%:0%-20%, 8 min) and then lyophilized under high vacuum to give compound 23 (7.3 mg, 17.2 umol, 7.55%yield, 95.4%purity) as a yellow solid.

LC-MS: 405.1

H NMR: (400 MHz, DMSO-d6) δ ppm 10.24 (br s, 1 H) , 8.76 (s, 1 H) , 8.33 -8.58 (m, 1 H) , 8.46 (br s, 1 H) , 7.69 (s, 1 H) , 7.51 (s, 1 H) , 7.20 -7.35 (m, 5 H) , 4.78 -4.90 (m, 1 H) , 4.62 (br s, 1 H) , 4.45 -4.54 (m, 1 H) , 4.09 (br s, 2 H) .

Compounds in Table 1 below were prepared by following procedures that are the same as or similar to those described above.

Table 1

Scheme IV below describes an illustrative synthetic scheme for preparation of compound 38 disclosed herein.

Scheme IV: Synthesis scheme for compound 38

General procedure for preparation of compound B-2

To a solution of compound B-1 (150 g, 955 mmol, 1.00 eq. ) in DMF (1000 mL) was added K ₂CO ₃ (198 g, 1.43 mol, 1.50 eq. ) . Then 3-bromoprop-1-ene (121 g, 1.00 mol, 1.05 eq. ) was added dropwise. The resulting mixture was stirred at 25 ℃ for 2 h. TLC (petroleum ether/EtOAc=10/1, UV) showed the starting material was consumed completely and a new spot with smaller polarity was formed. The mixture was diluted with EtOAc (1500 mL) , washed with H ₂O (1500 mL) , brine (300 mL*3) . The separated organic layer was dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give compound B-2 (188 g, 953 mmol, 99.8%yield) as a brown solid, which was directly used in the next step without further purification.

¹H NMR: (400 MHz, CDCl ₃) δ ppm 8.04 -8.14 (m, 1 H) , 6.70 -6.83 (m, 2 H) , 6.02 (ddt, J=17.27, 10.58, 5.27, 5.27 Hz, 1 H) , 5.33 -5.48 (m, 2 H) , 4.62 (dt, J=5.38, 1.47 Hz, 2 H)

General procedure for preparation of compound B-3

To a solution of Boc-L-Serine (125 g, 608 mmol, 1.00 eq. ) in DMF (1200 mL) was added NaH (49.9 g, 1.25 mol, 60%purity, 2.05 eq. ) in portions at 0 ℃under N ₂. The mixture was stirred at 0 ℃ for 30 minutes. Then a solution of compound B-2 (120 g, 608 mmol, 1.00 eq. ) in DMF (200 mL) was added dropwise. After the addition, the mixture was stirred at 25 ℃ for 1 h. The mixture was quenched by adding H ₂O (2000 mL) , acidified with 2 N of HCl to pH～5-6, extracted with EtOAc (1000 mL*2) . The combined extracts were dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give compound B-3 (230 g, crude) as brown oil, which was directly used in the next step without further purification.

LCMS: 470.

General procedure for preparation of compound B-4

A mixture of compound B-3 (220 g, crude) , NH ₄Cl (61.5 g, 1.15 mol, 2.00 eq.) in EtOH (1200 mL) and H ₂O (300 mL) was heated to 80 ℃. Then Fe (160 g, 2.88 mol, 5.00 eq. ) was added in portions. The mixture was stirred at 80 ℃ for 3 h. The mixture was filtered through a pad of celite and the filter cake was washed with MeOH (200 mL) . The combined filtrate was concentrated to give brown residue. The residue was stirred in DCM (500 mL) at 25 ℃ for 10 min. The mixture was filtered and the filtrate was concentrated to give compound B-4 (180 g, crude) as brown gum, which was directly used in the next step without further purification.

LC-MS: 353.1

General procedure for preparation of compound B-5

To a solution of compound B-4 (200 g, crude) in DCM (1000 mL) was added DIPEA (220 g, 1.70 mol, 296 mL, 3.00 eq. ) . Then T ₃P (361 g, 567 mmol, 337 mL, 50%purity, 1.00 eq. ) was added dropwise. The mixture was stirred at 25 ℃ for 2 h and then was concentrated and the residue was diluted with EtOAc (1500 mL) , washed with sat. NaHCO ₃ (2000 mL) , brine (500 mL) , dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give brown residue. The residue was purified by column chromatography (SiO ₂, petroleum ether/ethyl acetate=30/1 to 10/1) to give compound B-5 (120 g, 359 mmol, 21.1%yield) as a yellow solid.

¹H NMR (400 MHz, CDCl ₃) δ ppm 7.58 -7.80 (m, 1 H) , 6.90 (d, J=8.50 Hz, 1 H) , 6.64 -6.74 (m, 2 H) , 7.26 (s, 1 H) , 6.03 (ddt, J=17.26, 10.54, 5.30, 5.30 Hz, 1 H) , 5.52 (br d, J=5.00 Hz, 1 H) , 5.41 (dq, J=17.26, 1.50 Hz, 1 H) , 5.27 -5.33 (m, 1 H) , 4.58 -4.73 (m, 2 H) , 4.47 -4.54 (m, 2 H) , 4.20 (t, J=10.07 Hz, 1 H) , 1.41 -1.42 (m, 9 H)

General procedure for preparation of compound B-6

To a solution of compound B-5 (60.0 g, 80%purity, 143 mmol, 1.00 eq. ) in DMF (500 mL) was added K ₂CO ₃ (39.7 g, 287 mmol, 2.00 eq. ) . Then MeI (40.7 g, 287 mmol, 17.8 mL, 2.00 eq. ) was added at 25 ℃. The mixture was stirred at 25 ℃ for 18 h. TLC (petroleum ether/EtOAc=4/1, UV) showed the reaction was completed. The mixture was diluted with EtOAc (800 mL) , washed with water (1000 mL) , brine (200 mL*3) , dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give a crude product. The residue was purified by flash silica gel chromatography (

330 g

Silica Flash Column, Eluent of 0～15%Ethyl acetate/petroleum ether gradient @100 mL/min) to give compound B-6 (44.0 g, 126 mmol, 87.9%yield) as a yellow solid.

¹H NMR: (400 MHz, CDCl ₃) δ ppm 7.08 (d, J=8.80 Hz, 1 H) , 6.75 (dd, J=8.80, 2.69 Hz, 1 H) , 6.69 (d, J=2.69 Hz, 1 H) , 6.04 (ddt, J=17.21, 10.55, 5.26, 5.26 Hz, 1 H) , 5.50 (br d, J=6.97 Hz, 1 H) , 5.38 -5.46 (m, 1 H) , 5.31 (dd, J=10.51, 1.34 Hz, 1 H) , 4.45 -4.72 (m, 4 H) , 4.09 -4.18 (m, 1 H) , 3.35 (s, 3 H) , 1.39 (s, 9 H)

General procedure for preparation of compound B-6A

To a solution of compound B-6 (44.0 g, 126 mmol, 1.00 eq. ) in DCM (400 mL) was added dropwise TFA (100 mL) at 25 ℃. The mixture was stirred at 25 ℃for 1 h. The mixture was concentrated to give compound B-6A (45.7 g, crude, TFA salt) as brown gum, which was directly used in the next step without further purification.

LC-MS: 249.0

General procedure for preparation of compound B-7

To a solution of compound B-6A (45.7 g, TFA salt, 126 mmol, 1.00 eq. ) in DCM (400 mL) was added DIPEA (48.9 g, 378 mmol, 65.9 mL, 3.00 eq. ) . Then TFAA (31.8 g, 151 mmol, 21.1 mL, 1.20 eq. ) was added under N ₂ at 0 ℃. After the addition, the mixture was stirred at 0 ℃ for 20 min. The mixture was diluted with DCM (100 mL) , washed with brine (200 mL) , dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give compound B-7 (41.0 g, 119 mmol, 94.4%yield) as a yellow solid, which was directly used in the next step without further purification.

LC-MS: 345.0

¹H NMR: (400 MHz, CDCl ₃) δ ppm 7.53 (br s, 1 H) , 7.11 (d, J=8.88 Hz, 1 H) , 6.81 (dd, J=8.88, 2.88 Hz, 1 H) , 6.75 (d, J=2.75 Hz, 1 H) , 6.05 (ddt, J=17.26, 10.54, 5.30, 5.30 Hz, 1 H) , 5.44 (dq, J=17.26, 1.54 Hz, 1 H) , 5.33 (dq, J=10.52, 1.37 Hz, 1 H) , 4.79 -4.88 (m, 1 H) , 4.72 (dd, J=9.63, 7.38 Hz, 1 H) , 4.52 -4.57 (m, 2 H) , 4.19 (dd, J=10.94, 9.69 Hz, 1 H) , 3.39 (s, 3 H)

General procedure for preparation of compound B-8

A solution of compound B-7 (40.0 g, 116 mmol, 1.00 eq. ) in TFAA (1000 mL) was cooled to -30 ℃ under N ₂. Then HNO ₃ (11.8 g, 127 mmol, 8.46 mL, 68%purity, 1.10 eq. ) was added dropwise while keeping the inter temperature around -30 ℃. After the addition, the mixture was stirred at -30 ℃ for 1 h. TLC (petroleum ether/EtOc=5/1, UV) showed the starting material was consumed completely. The mixture was poured onto ice water (2500 mL) slowly, solid precipitated. The solid was collected by filtration, dried under vacuum to give compound B-8 (43.5 g, 112 mmol, 96.2%yield) as a yellow solid, which was directly used in the next step without further purification.

¹H NMR: (400 MHz, CDCl ₃) δ ppm 7.79 -7.93 (m, 1 H) , 7.51 (br d, J=5.27 Hz, 1 H) , 6.83 -6.99 (m, 1 H) , 6.83 -6.99 (m, 1 H) , 5.95 -6.16 (m, 1 H) , 5.54 (br d, J=17.32 Hz, 1 H) , 5.40 (d, J=10.54 Hz, 1 H) , 4.82 -4.93 (m, 1 H) , 4.66 -4.80 (m, 3 H) , 4.26 -4.37 (m, 1 H) , 3.44 (s, 3 H)

General procedure for preparation of compound B-9

To a solution of compound B-8 (43.0 g, 88.3 mmol, 1.00 eq. ) in DCM (1000 mL) was added Pd (PPh ₃) ₄ (40.8 g, 35.3 mmol, 0.40 eq. ) , Bu ₃SnH (77.1 g, 265 mmol, 70.1 mL, 3.00 eq. ) and AcOH (31.8 g, 530 mmol, 30.3 mL, 6.00 eq. ) under N ₂. The mixture was stirred at 25 ℃ for 1 h. TLC (petroleum ether/EtOAc=3/1, UV) showed the reaction was completed. The mixture was concentrated and the residue was triturated in petroleum ether (1500 mL) , filtered and the filtrate was discarded. The filter cake was dried to give black residue. The residue was purified by flash silica gel chromatography (

330 g

Silica Flash Column, Eluent of 0 ～ 1%MeOH/DCM gradient @100 mL/min) to give compound B-9 (21.3 g, 59.2 mmol, 67.0%yield, 97.1%purity) as a yellow solid.

LCMS: 350.1

¹H NMR: (400 MHz, CDCl ₃) δ ppm 10.62 (br s, 1 H) , 8.02 (s, 1 H) , 7.47 (br s, 1 H) , 6.99 (s, 1 H) , 4.80 -4.94 (m, 1 H) , 4.72 (dd, J=9.90, 6.85 Hz, 1 H) , 4.31 (dd, J=11.25, 9.90 Hz, 1 H) , 3.46 (s, 3 H)

General procedure for preparation of compound B-10

To a solution of compound B-9 (860 mg, 2.46 mmol, 1 eq) in MeOH (40 mL) was added Boc ₂O (537 mg, 2.46 mmol, 565 uL, 1 eq) and Pd/C (0.1 g, 10%purity, 1.00 eq) under N ₂. The mixture was degassed and purged with H ₂ for 3 times. Then the mixture was stirred under H ₂ (15 PSI) at 25 ℃ for 18 h. The mixture was filtered through a pad of celite and the filtrate was concentrated to give compound B-10 (940 mg, 2.24 mmol, 91.0%yield) as brown gum.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 10.21 (br s, 1 H) , 9.75 (br d, J=7.95 Hz, 1 H) , 7.96 (s, 1 H) , 7.64 (s, 1 H) , 6.66 (s, 1 H) , 4.72 (dt, J=11.31, 7.92 Hz, 1 H) , 4.46 (t, J=10.70 Hz, 1 H) , 4.33 -4.41 (m, 1 H) , 4.11 (q, J=5.22 Hz, 1 H) , 3.22 (s, 3 H) , 3.18 (d, J=5.26 Hz, 2 H) , 1.47 -1.47 (m, 9 H)

LCMS: [M+H-56] ⁺ = 364.0, RT = 0.897 min

General procedure for preparation of compound B-11

A solution of B-10 (500 mg, 1.19 mmol, 1 eq) in THF (5 mL) and H ₂O (5 mL) was added K ₂CO ₃ (247 mg, 1.79 mmol, 1.5 eq) . After the addition, the mixture was stirred at 25 ℃ for 18 h. LCMS showed the starting material was remained and the desired product was formed. The mixture was stirred at 50 ℃ for another 42 h and the was concentrated to give B-11 (400 mg, crude) as a yellow solid, which was directly used in the next step without further purification.

LCMS: [M+H] ⁺ = 324.1, RT = 0.801 min

General procedure for preparation of compound B-12

To a mixture of B-11 (330 mg, 1.02 mmol, 1 eq) , 5-benzyl-4H-1, 2, 4-triazole-3-carboxylic acid (248 mg, 1.22 mmol, 1.2 eq) in DMF (5 mL) was added EDCI (234.78 mg, 1.22 mmol, 1.2 eq) and 1-oxidopyridin-1-ium-2-ol (136 mg, 1.22 mmol, 1.2 eq) and TEA (309 mg, 3.06 mmol, 426 uL, 3 eq) . The mixture was stirred at 50 ℃ for 1 h. The mixture was concentrated to give yellow residue. The residue was purified by flash silica gel chromatography (

10 g

Silica Flash Column, Eluent of 0～2%MeOH/DCM gradient @35 mL/min) to give compound B-12 (250 mg, 491 umol, 48.2%yield) as a yellow solid.

LCMS: [M+Na] ⁺ = 531.1, RT = 0.911 min

General procedure for preparation of compound B-13

To a solution of B-12 (200 mg, 393 umol, 1 eq) in EtOAc (1 mL) was added HCl/MeOH (4 M, 5 mL, 50.85 eq) . The mixture was stirred at 25 ℃ for 1 h. The mixture turned yellow clear. LCMS showed the starting material was remained. The mixture was stirred at 25 ℃ for another 17 h and then was concentrated to give a purple solid (crude, 170 mg) . A suspension of crude product (30 mg, 67.4 umol, 1 eq, HCl salt) in EtOAc (5 mL) was stirred at 25 ℃ for 1 h. The mixture was filtered and the filter cake was dried under vacuum to give B-13 (6.2 mg, 15.18 umol, 22.5%yield) as a purple solid.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 10.91 (s, 1 H) , 8.42 (s, 1 H) , 7.20 -7.39 (m, 6 H) , 6.81 (s, 1 H) , 4.78 -4.90 (m, 1 H) , 4.54 (t, J=11.19 Hz, 1 H) , 4.31 -4.45 (m, 1 H) , 4.12 (s, 2 H) , 3.23 (s, 3 H)

LCMS: 409.1;

General procedure for preparation of compound 38

To a solution of compound B-13 (100 mg, 245 umol, 1 eq) in CHCl ₃ (3 mL) was added CH ₃COOH (58.8 mg, 979 umol, 56.0 uL, 4 eq) and pyrrolidine (34.8 mg, 490 umol, 40.9 uL, 2 eq) C (OMe) ₄ (66.7 mg, 490 umol, 2 eq) . The mixture was stirred at 60 ℃ for 6 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Gemini-NX 150*30mm*5um; mobile phase: [water (0.05%NH ₃H ₂O+10mM NH ₄HCO ₃) -ACN] ; B%: 30%-50%, 8 min) . Compound 38 (0.03 g, 60.9 umol, 24.9%yield, 99%purity) was obtained as a white solid.

LC-MS: 244.6

¹H NMR (400MHz, DMSO-d ₆) δ = 8.58 -8.27 (m, 1H) , 8.43 (br s, 1H) , 7.41 (s, 1H) , 7.37 (s, 1H) , 7.35 -7.30 (m, 2H) , 7.29 -7.23 (m, 3H) , 4.91 -4.73 (m, 1H) , 4.53 (t, J = 10.6 Hz, 1H) , 4.39 (dd, J = 7.9, 9.8 Hz, 1H) , 4.12 (s, 2H) , 3.56 (br t, J = 6.3 Hz, 4H) , 3.32 (s, 3H) , 2.04 -1.92 (m, 4H) .

Scheme V below describes an illustrative synthetic scheme for preparation of compound 52 disclosed herein.

Scheme V: Synthesis scheme for compound 52:

General procedure for preparation of compound B-14

To a solution of compound B-9 (2.12 g, 6.07 mmol, 1.00 eq. ) in MeOH (50 mL) was added Pd/C (0.30 g, 10%purity) under N ₂. The mixture was degassed and purged with H ₂ for 3 times. Then the mixture was stirred at 25 ℃ under H ₂ (15 psi) for 2 h. LC-MS showed the reaction was complete. The mixture was filtered through a pad of celite and the filtrate was concentrated to give compound B-14 (1.90 g, 5.95 mmol, 98.0%yield) as a yellow solid, which was directly used in the next step without further purification.

¹H NMR: (400 MHz, DMSO-d ₆) δ ppm 9.70 (br s, 1 H) , 9.46 (br s, 1 H) , 6.60 (s, 1 H) , 6.48 (s, 1 H) , 4.51 -4.78 (m, 3 H) , 4.34 -4.42 (m, 1 H) , 4.23 -4.31 (m, 1 H) , 3.17 (s, 3 H)

LC-MS: 320.1

General procedure for preparation of compound B-15

To a stirred solution of compound B-14 (210 mg, 658 umol, 1.00 eq. ) in DCM (2 mL) and MeOH (2 mL) was added BrCN (139 mg, 1.32 mmol, 96.7 uL, 2.00 eq. ) . The resulting solution was stirred at 25 ℃ for 16 h. LCMS indicated start material was consumed and the desired product was observed. The mixture was filtered and concentrated to give a brown residue. To the residue was added 5 mL water and extracted with 5 mL EtOAc for 3 times. The organic layer was dried over Mg ₂SO ₄, filtered and concentrated. The residue was purified by flash silica gel chromatography (

4 g

Silica Flash Column, Eluent of 0 ～ 80%Ethyl acetate/petroleum ether gradient @18 mL/min) to give B-15 (95.7 mg, 278 umol, 42.2%yield) as a gray solid.

¹H NMR: (400 MHz, DMSO-d ₆) δ ppm 9.75 (s, 1 H) , 7.54 (s, 1 H) , 7.34 (s, 1 H) , 7.27 (s, 1 H) , 4.73 -4.68 (m, 1 H) , 4.51 –4.46 (m, 1 H) , 4.41-4.39 (m, 1 H) , 3.30 (s, 3 H)

LCMS: 344.9

General procedure for preparation of compound 52

To a solution of 5- [ (2-chlorophenyl) methyl] -1-methyl-1, 2, 4-triazole-3-carboxylic acid (101 mg, 402 umol, 1 eq) in DMF (5 mL) was added HOBt (59.8 mg, 443 umol, 1.1 eq) , EDCI (84.9 mg, 443 umol, 1.1 eq) and DIPEA (156 mg, 1.21 mmol, 210 uL, 3 eq) . The mixture was stirred for 10 min, then B-16 (100 mg, 402 umol, 1 eq) was added. The resulting mixture was stirred at 50 ℃ for 1 h. The mixture was poured into water (20 mL) , extracted with EtOAc (20 mL*2) . The combined EtOAc layer was dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give a crude product. The crude product was purified by prep-HPLC (column: YMC Triart C18 250*50mm*7um; mobile phase: [water (0.05%ammonia hydroxide v/v) -ACN] ; B%: 18%-58%, 10 min) to give compound 52 (78.9 mg, 157 umol, 39.1%yield, 96.2%purity) as a yellow solid.

LCMS: 482.1

¹H NMR: (400 MHz, DMSO-d ₆) δ ppm 8.23 (d, J=8.03 Hz, 1 H) , 7.52 (s, 2 H) , 7.48 (dd, J=5.65, 3.64 Hz, 1 H) , 7.30 -7.37 (m, 3 H) , 7.24 -7.29 (m, 2 H) , 4.80 (dt, J=11.11, 8.00 Hz, 1 H) , 4.47 (t, J=10.67 Hz, 1 H) , 4.35 -4.40 (m, 1 H) , 4.31 (s, 2 H) , 3.90 (s, 3 H) , 3.30 (s, 3 H)

Compounds in Table 2 below were prepared by following procedures that are the same as or similar to those for the preparation of compounds 38 and 52 described above.

Table 2

Scheme VI below describes an illustrative synthetic scheme for preparation of compound 70 disclosed herein.

Scheme VI: Synthesis scheme for compound 70

General procedure for preparation of compound C-2

To a stirred solution of NaH (28.0 g, 700 mmol, 60%purity, 2.2 eq) in DMF (500 mL) was added a solution of Boc-L-serine (71.8 g, 350 mmol, 1.1 eq) in DMF (400 mL) drop wised at 0 ℃ under N ₂ over 0.5 h . The mixture was stirred at 0 ℃for 0.5 h till no more H ₂ released. Then to the mixture was added the suspension of compound C-1 (70 g, 318 mmol, 1 eq) in DMF (100 mL) at 0 ℃ dropwise over 0.5 h. After addition, the mixture was allowed to warm to 20 ℃ over a period of 2 h under N ₂ to give a dark brown suspension. The reaction mixture was cooled to 0 ℃, poured into ice water (2000 mL) slowly, acidified with 2N HCl (～500 mL) to pH=3, extracted with EtOAc (2000 mL *2) . The combined organic layers were washed with brine (1500 mL) , dried over Na ₂SO ₄, filtered and concentrated under reduced pressure to give as a brown oil. The residue was used for next step without further purification. Compound C-2 (110 g, crude) was obtained as a brown oil.

General procedure for preparation of compound C-3

To a stirred solution of compound C-2 (100 g, 247 mmol, 1 eq) in CH ₃COOH (1000 mL) was added Zn (64.6 g, 987 mmol, 4 eq) in potions at 25 ℃over 2 h. The mixture was stirred at 25 ℃ for 15 h. The reaction mixture was filter through a pad of celite, washed with DCM. The filter cake was quenched by HCl (1500mL, 3 N) . The liquid phase was partitioned between water (2000 mL) and ethyl acetate (4000 mL) . The organic layers were washed with saturated solution of NaCl in water (2000 m) , dried over MgSO ₄, filtered and concentrated under reduced pressure to give compound C-3 (80 g, crude) as a brown oil which was used to next step without further purification.

General procedure for preparation of compound C-4

To a solution of compound C-3 (70 g, 187 mmol, 1 eq) in THF (600 mL) was added DIEA (97 g, 746 mmol, 130 mL, 4 eq) and T ₃P (119 g, 187 mmol, 111 mL, 50%purity, 1 eq) . The mixture was stirred at 25 ℃ for 5 h. The reaction mixture was partitioned between saturated solution of NaHCO ₃ in water (200 mL) and ethyl acetate (600 mL) . The organic phase was separated dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chomatography (

12 g

Silica Flash Column, Eluent of 25%Ethyl acetate/Petroleum ether gradient @20 mL/min) to afford compound C-4 (32 g, 90.0 mmol, 48.0%yield) a white solid.

LCMS: 300.9

¹H NMR (400MHz, CDCl ₃) : δ = 8.14 (br s, 1H) , 7.32 (d, J=2.0 Hz, 1H) , 7.25 (dd, J=2.1, 8.4 Hz, 1H) , 6.91 (d, J=8.5 Hz, 1H) , 5.53 (br d, J=6.0 Hz, 1H) , 4.79 -4.57 (m, 2H) , 4.41 -4.06 (m, 1H) , 1.46 (s, 9H)

General procedure for preparation of compound C-5:

To a solution of compound C-4 (22 g, 58.5 mmol, 648 uL, 95%purity, 1 eq) in DMF (200 mL) was added K ₂CO ₃ (8.10 g, 58.5 mmol, 1 eq) MeI (12.5 g, 87.8 mmol, 5.46 mL, 1.5 eq) at 0 ℃. The mixture was stirred at 25 ℃ for 5 h. The reaction mixture was quenched by addition water (500 mL) at 25 ℃, and then extracted with ethyl acetate (500mL *2) . The combined organic layers were dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

220 g

Silica Flash Column, Eluent of 10%Ethyl acetate/Petroleum ether gradient @100 mL/min) . Compound C-5 (19 g, 48.6 mmol, 83.1%yield, 95%purity) was obtained as a white solid.

¹H NMR: (400MHz, CDCl ₃) δ= 7.38 -7.33 (m, 2H) , 7.09 (d, J=8.5 Hz, 1H) , 5.50 (br d, J=6.8 Hz, 1H) , 4.73 -4.62 (m, 1H) , 4.61 -4.46 (m, 1H) , 4.20 (dd, J=9.4, 10.9 Hz, 1H) , 3.40 (s, 3H) , 1.42 (s, 9H)

General procedure for preparation of compound C-6:

To a solution of compound C-5 (19 g, 51.2 mmol, 1 eq) in DCM (100 mL) was added CH ₃COOH (24.6 g, 409 mmol, 23.4 mL, 8 eq) at 25 ℃. The mixture was stirred at 25 ℃ for 5 h. The reaction mixture was concentrated under reduced pressure to give compound C-6 (20 g, 46.7 mmol, 91.3%yield, 90%purity, TFA salt) as a brown solid which was used to next step without further purification.

General procedure for preparation of compound C-7:

To a stirred solution of compound C-6 (20 g, 51.9 mmol, 1 eq, TFA) in DCM (200 mL) was added DIEA (23.5 g, 182 mmol, 31.7 mL, 3.5 eq) and TFAA (15.3 g, 72.7 mmol, 10.1 mL, 1.4 eq) . The mixture was stirred at 25 ℃ for 3 h. The reaction mixture was partitioned between water (50 ml) and DCM (100 mL) , dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

220 g

Silica Flash Column, Eluent of 10%Ethyl acetate/Petroleum ether gradient @100mL/min) to afford compound C-7 (17 g, 44 mmol, 84.7%yield, 95%purity) as a light-yellow solid.

General procedure for preparation of compound C-8:

To a solution of compound C-7 (12.0 g, 32.7 mmol, 1 eq) in TFAA (1000 mL) was added HNO ₃ (4.44 g, 45.8 mmol, 3.17 mL, 65%purity, 1.4 eq) at -30 ℃in drop wised. The mixture was cooled to -30 ℃ and stirred for 5 h. The reaction mixture was poured to ice water, filtered. The filter cake was rinsed with water (50 mL) , dried to give compound C-8 (10 g, 14.6 mmol, 44.5%yield, 60%purity) as a white solid which was used to next step without further purification.

General procedure for preparation of compound C-9:

To a solution of compound C-8 (2 g, 4.83 mmol, 1 eq) in dioxane (20 mL) was added palladium; triphenylphosphane (279 mg, 241 umol, 0.05 eq) and tributyl (1-ethoxyvinyl) stannane (1.92 g, 5.31 mmol, 1.79 mL, 1.1 eq) . The mixture was stirred at 100 ℃ for 18 h. The mixture was cooled to 25 ℃, HCl (6N, 2eq) was added, the resulting mixture was stirred at 25 ℃ for 0.5 h. The mixture was extracted with ethyl acetate (100 mL) . The organic layer was concentrated to give a residue. The residue was purified by flash silica gel chromatography (

12g

Silica Flash Column, Eluent of 20%Ethyl acetate/Petroleum ether gradient @20 mL/min) to afford compound C-9 (1.7 g, 3.60 mmol, 74.6%yield, 80%purity) as a yellow solid.

¹H NMR: (400MHz, CHLOROFORM-d) δ = 8.03 (s, 1H) , 7.49 (br d, J=5.5 Hz, 1H) , 7.25 (s, 1H) , 4.89 (dd, J=6.4, 11.3 Hz, 1H) , 4.85 -4.78 (m, 1H) , 4.41 (dd, J=9.8, 11.1 Hz, 1H) , 3.53 (s, 3H) , 2.61 (s, 3H)

General procedure for preparation of compound C-10:

To a solution of compound C-9 (4 g, 9.71 mmol, 1 eq) in ethanol (40 mL) , Water (40 mL) was added NH ₄Cl (5.19 g, 97.0 mmol, 10 eq) and Fe (3.79 g, 68.0 mmol, 7 eq) . The mixture was stirred at 90 ℃ for 5 h. The reaction mixture was filtered and partitioned between water (100 mL) and ethyl acetate (200 mL) . The organic phase was separated, washed with saturated solution of NaCl in water (50 mL) , dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

12 g

Silica Flash Column, Eluent of 30%Ethyl acetate/Petroleum ether gradient @30 mL/min) . Compound C-10 (1.2 g, 2.98 mmol, 30.7%yield, 95%purity) was obtained as a white solid.

General procedure for preparation of compound C-11:

To a suspension of compound C-10 (3.5 g, 10.1 mmol, 1 eq) in CH ₃COOH (60 mL) was added potassium; cyanate (4.11 g, 50.7 mmol, 2.00 mL, 5 eq) . The mixture was stirred at 25 ℃ for 1 h and 60 ℃ for 3 h. The residue was diluted with water (100 mL) and extracted with ethyl acetate (100 mL *2) . The combined organic layers were washed with brine (200 mL) , dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

12g

Silica Flash Column, Eluent of 50%THF/DCM gradient @30mL/min) . Compound C-11 (1.2 g, 3.08 mmol, 30.4%yield, 95%purity) was obtained as a yellow solid.

LCMS: 388.0

¹H NMR (400MHz, CDCl ₃) δ = 13.21 (br s, 1H) , 7.74 (s, 1H) , 7.54 (br d, J=6.0 Hz, 1H) , 7.33 (s, 1H) , 4.96 -4.75 (m, 2H) , 4.27 (t, J=10.0 Hz, 1H) , 3.57 (s, 3H) , 2.87 (s, 3H)

General procedure for preparation of compound C-12:

To a solution of compound C-11 (0.22 g, 594 umol, 1 eq) in H ₂O (5 mL) EtOH (5 mL) was added K ₂CO ₃ (246 mg, 1.78 mmol, 3 eq) . The mixture was stirred at 25 ℃ for 5 h. The reaction mixture was lyophilized to give a solid. The solid was used to next step without further purification. Compound C-12 (0.470 g, 565 umol, 95.2%yield, 33%purity) was obtained as a yellow solid.

General procedure for preparation of compound 70

To a solution of compound C-12 (0.470 g, 565 umol, 33.0%purity, 1 eq) 5-benzyl-4H-1, 2, 4-triazole-3-carboxylic acid (115 mg, 565 umol, 1 eq) in DMF (5 mL) was added T ₃P (720 mg, 1.13 mmol, 672 uL, 50%purity, 2 eq) and DIEA (365 mg, 2.83 mmol, 492 uL, 5 eq) . The mixture was stirred at 25 ℃ for 1 h. The residue was diluted with water (100 mL) and extracted with ethyl acetate (100 mL *2) . The combined organic layers were washed with water (200 mL) , dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: YMC Triart C18 250*50mm*7um; mobile phase: [water (0.225%FA) -ACN] ; B%: 4%-44%, 9min) . And then re-purified by prep-HPLC (column: Phenomenex Gemini-NX C18 75*30mm*3um; mobile phase: [water (0.05%ammonia hydroxide v/v) -ACN] ; B%: 0%-40%, 9 min) . Compound 70 (0.046 g, 99.2 umol, 17.5%yield, 99.1%purity) was obtained as a yellow solid.

LCMS: 460.1

¹H NMR (400MHz, DMSO-d6) δ = 12.10 -11.53 (m, 1H) , 8.58 (br d, J=5.3 Hz, 1H) , 7.83 (s, 1H) , 7.36 -7.16 (m, 6H) , 4.89 (td, J=7.9, 11.5 Hz, 1H) , 4.62 (t, J=10.8 Hz, 1H) , 4.44 (dd, J=7.9, 9.7 Hz, 1H) , 4.12 (s, 2H) , 3.35 (s, 3H) , 2.67 (s, 3H)

Compounds in Table 3 below were prepared by following procedures that are the same as or similar to those for the preparation of compound 70 described above.

Table 3

Scheme VII below describes an illustrative synthetic scheme for preparation of compound 87 disclosed herein.

Scheme VII: Synthesis scheme for compound 87

General procedure for preparation of compound D-2

To a stirred solution of NaNO ₃ (5.2 g, 61.1 mmol) in Conc. H ₂SO ₄ (60 mL) cooled to 0 ℃ was added 6-chloroquinoline D-1 (10.0 g, 61.1 mmol) in Conc. H ₂SO ₄ (60 mL) dropwise. After addition the solution was stirred at 0 ℃ for 2 h. The reaction mixture was poured into stirred cold water (600 mL) , then extracted with ethyl acetate (2X500 mL) . The combined organic extracts were washed with saturated sodium bicarbonate solution (400 mL) and brine (400 mL) , dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel chromatography (ethyl acetate/hexanes) to afford 6-chloro-5-nitroquinoline D-2 (11.2 g, 87%yield) .

¹H NMR (500 MHz, CDCl ₃) δ 9.01 (dd, J = 4.2, 1.6 Hz, 1H) , 8.20 (dd, J = 9.1, 0.6 Hz, 1H) , 8.05 (ddd, J = 8.7, 1.4, 0.9 Hz, 1H) , 7.76 (d, J = 9.1 Hz, 1H) , 7.59 (dd, J =8.7, 4.2 Hz, 1H) .

¹³C NMR (125 MHz, CDCl ₃) δ 152.04, 146.31, 145.59, 133.49, 130.29, 129.92, 124.23, 124.01, 121.21.

General procedure for preparation of compound D-3

To a stirred solution of NaH (3.1 g) in anhydrous DMF (20 mL) cooled to -5 ℃ was added (tert-butoxycarbonyl) -L-serine (7.9 g) in anhydrous DMF (50 mL) dropwise. After addition the solution was stirred at -5 ℃ for 15 min. 6-chloro-5-nitroquinoline D-2 (5 g) in anhydrous DMF (30 mL) was added dropwise. The reaction mixture was stirred at -5 ℃ for another 5 h before was poured into cold water (300 mL) . Extracted with ethyl acetate (2X200 mL) after the pH was adjusted to 4. The combined organic extract was washed with water (2X300 mL) and brine (400 mL) , dried over sodium sulfate and concentrated under reduced pressure to afford crude compound D-3 (8.7 g) as brown oil.

MS-ESI: 336 [M-H] ^-.

General procedure for preparation of compound D-4

To a solution of compound D-3 in ethyl acetate/ethyl alcohol (1: 1) was added Pd-C (1.7 g) . The reaction mixture was stirred at room temperature for 5 h under H ₂. After the reaction is completed, the reaction mixture was filtrated. The filtrate was concentrated under reduced pressure to afford crude compound D-4 (8 g) .

MS-ESI: 348 [M+H] ⁺

General procedure for preparation of compound D-5

To a stirred solution of crude compound D-4 (8 g) in anhydrous THF (200 mL) were added DIPEA (4.5 g) and HATU (10.5 g) . The reaction mixture was stirred at room temperature for 4 h. 10 mL water was added after the reaction was completed, and the volatiles were removed under reduced pressure. The residue was added into ethyl acetate and extracted with water. The organic extracts were washed with brine (400 mL) , dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel chromatography (ethyl acetate/hexanes) to afford tert-butyl (S) - (2-oxo-1, 2, 3, 4-tetrahydro- [1, 4] oxazepino [3, 2-f] quinolin-3-yl) carbamate (compound D-5, 3 g, 36%yield for 3 step) .

MS-ESI: 330 [M+H] ⁺

General procedure for preparation of compound D-6

To a stirred solution of compound 5 (3 g) in DMF (10 mL) cooled into -5 ℃ was added Cs ₂CO ₃ (4 g) . After addition the reaction mixture was stirred for 5 min at -5 ℃. CH ₃I (1.4 g) in DMF was added dropwise. After the addition completed, the reaction mixture was stirred for another 5 h at -5 ℃. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (2X80 mL) . The combined organic extract were washed with water (2X150 mL) and brine (150 mL) , dried over sodium sulfate and concentrated under reduced pressure to afford tert-butyl (S) - (1-methyl-2-oxo-1, 2, 3, 4-tetrahydro- [1, 4] oxazepino [3, 2-f] quinolin-3-yl)carbamate (Compound D-6, 2.9 g) .

MS-ESI: 344 [M+H] ⁺.

General procedure for preparation of compound 87

To a solution of compound D-6 (100 mg) in EtOH (3 mL) was added concentrated hydrochloric acid (1 mL) . The reaction mixture was stirred at room temperature for 2 h. The volatiles were removed under reduced pressure to get a yellow solid.

To a solution of D-7 (95 mg) in DMF (1 mL) was added DIPEA (152mg) , EDCI (82 mg) , and HOBt (58 mg) . The reaction mixture was stirred at room temperature for 30 min. A solution of above-mentioned yellow solid in DMF (1 mL) was added into the reaction mixture. After addition the reaction was stirred over night at room temperature. The reaction mixture was diluted with water (15 mL) and extracted with ethyl acetate (2X15 mL) . The combined organic extracts were washed with water (2X30 mL) and brine (30 mL) , dried over sodium sulfate and concentrated under reduced pressure. The residue was purified via silica gel chromatography (MeOH/DCM) to afford (S) -3- (2-chlorobenzyl) -N- (1-methyl-2-oxo-1, 2, 3, 4-tetrahydro- [1, 4] oxazepino [3, 2-f] quinolin-3-yl) -1H-1, 2, 4-triazole-5-carboxamide (compound 87, 83 mg, 62%yield) .

¹H NMR (500 MHz, CD ₃OD) δ 9.12 (d, J = 4.9 Hz, 1H) , 8.93 (d, J = 8.6 Hz, 1H) , 8.19 (d, J = 9.1 Hz, 1H) , 7.98 (dd, J = 8.7, 5.0 Hz, 1H) , 7.90 (d, J = 9.1 Hz, 1H) , 7.38 (dd, J = 5.5, 3.8 Hz, 1H) , 7.34 (dd, J = 5.5, 3.8 Hz, 1H) , 7.29-7.22 (m, 2H) , 5.13-5.06 (m, 1H) , 4.72 (dd, J = 10.0, 7.6 Hz, 1H) , 4.66-4.58 (m, 1 H) , 4.29 (s, 2H) , 3.45 (s, 3H) .

¹³C NMR (125 MHz, CD ₃OD) δ 170.87, 159.18, 150.68, 146.19, 139.77, 138.93, 133.73, 133.58, 131.41, 131.06, 129.31, 129.19, 128.90, 128.76, 128.61, 127.11, 125.24, 124.34, 122.09, 77.51, 48.84, 36.73, 30.25.

MS-ESI: 462 [M+H] ⁺.

Compounds in Table 4 below were prepared by following procedures that are the same as or similar to those for the preparation of compound 87 described above.

Table 4

Scheme VIII below describes an illustrative synthetic scheme for preparation of compound 133 disclosed herein.

Scheme VIII: Synthesis scheme for compound 133

General procedure for preparation of compound E-2:

To a solution of compound E-1 (30.0 g, 173 mmol, 1.00 eq) , Boc-L-serine (39.9 g, 182 mmol, 1.05 eq) , Cu (OAc) ₂ (3.15 g, 17.3 mmol, 0.10 eq) , DMAP (4.24 g, 34.7 mmol, 0.20 eq) in DCM (250 mL) was added DMAP (4.24 g, 34.7 mmol, 0.20 eq) . The reaction mixture was stirred at 20 ℃ for 16 hrs. The residue was poured into water (250 mL) . The aqueous phase was extracted with DCM (250 mL*2) . The combined organic phase was washed with brine (250 mL*1) , dried with anhydrous Na ₂SO ₄, filtered and concentrated in vacuum. The residue was purified by flash silica gel chromatography (

120 g

Silica Flash Column, Eluent of 0～20%Ethyl acetate/Petroleum ether gradient @100 mL/min) to give compound E-2 (20.0 g, 17.3 mmol, 9.99%yield, 30%purity) as a yellow oil. ¹H NMR: (400 MHz, DMSO-d6) δ ppm 8.62 (d, J=2.88 Hz, 1 H) , 8.38 (dd, J=8.32, 0.94 Hz, 2 H) , 7.93 -7.98 (m, 1 H) , 7.86 -7.92 (m, 1 H) , 7.59 -7.65 (m, 3 H) , 4.54 -4.65 (m, 1 H) , 4.35 -4.45 (m, 2 H) , 3.70 (s, 3 H) , 1.40 (br s, 9 H)

General procedure for preparation of compound E-3:

To a solution of compound E-2 (20.0 g, 17.3 mmol, 322 uL, 30%purity, 1.00 eq) in DCM (150 mL) was added m-CPBA (7.03 g, 34.6 mmol, 85%purity, 2.00 eq) . The solution was stirred at 20 ℃ for 16 hrs. The reaction mixture was poured into water (200 mL) . The solution was extracted with DCM (200 mL*3) . The combined organic phase was washed with brine (200 mL*2) , dried with anhydrous Na ₂SO ₄, filtered and concentrated in vacuum. The residue was purified by flash silica gel chromatography (

220 g

Silica Flash Column, Eluent of 0～100%Ethyl acetate/Petroleum ether gradient @80 mL/min) to afford compound E-3 (3.40 g, 9.38 mmol, 54.2%yield) as a yellow oil.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 8.37 -8.45 (m, 2 H) , 7.95 (d, J=8.25 Hz, 1 H) , 7.60 -7.70 (m, 2 H) , 7.51 -7.57 (m, 2 H) , 4.48 -4.59 (m, 1 H) , 4.31 -4.46 (m, 2 H) , 3.68 (s, 3 H) , 1.34 -1.42 (m, 1 H) , 1.31 -1.42 (m, 8 H) .

LC-MS: 363.2

General procedure for preparation of compound E-4:

To a solution of compound E-3 (3.40 g, 9.38 mmol, 322 uL, 1.00 eq) and NH ₃.H ₂O (15.4 g, 132 mmol, 17.0 mL, 30%purity, 14.1 eq) in DCM (45 mL) was added 4-methylbenzenesulfonyl chloride (3.58 g, 18.8 mmol, 2.00 eq) . The reaction mixture was stirred at 20 ℃ for 16 hrs. The reaction mixture was extracted with DCM (20 mL*2) . The combined organic phase was dried with anhydrous Na ₂SO ₄, filtered and concentrated in vacuum. The residue was purified by flash silica gel chromatography (

40 g

Silica Flash Column, Eluent of 0～40～100%Ethyl acetate/Petroleum ether gradient @40 mL/min) to afford compound E-4 (1.50 g, 4.15 mmol, 44.2%yield) as a yellow oil.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 7.85 (d, J=9.17 Hz, 1 H) , 7.58 (d, J=7.09 Hz, 1 H) , 7.41 -7.46 (m, 1 H) , 7.31 -7.37 (m, 2 H) , 7.10 -7.18 (m, 1 H) , 6.52 (br s, 2 H) , 4.64 -4.74 (m, 1 H) , 4.52 (dd, J=9.90, 4.03 Hz, 1 H) , 4.16 (dd, J=9.84, 3.36 Hz, 1 H) , 3.67 (s, 3 H) , 1.43 (s, 9 H)

General procedure for preparation of compound E-5:

To a solution of compound E-4 (1.50 g, 4.15 mmol, 1.00 eq) in THF (30 mL) and H ₂O (10 mL) was added LiOH. H ₂O (139 mg, 3.32 mmol, 0.80 eq) at 0 ℃. The reaction mixture was stirred at 0 ℃ for 1 hr. The reaction mixture was concentrated under reduced pressure at 20 ℃ to 10 mL. Solid was precipitated out. EtOAc (20 mL) and water (10 mL) was added in it and the suspension was stirred for 10 mins. The suspension was filtered and the residue was concentrated under reduced pressure to afford compound E-5 (0.7 g, 2.02 mmol, 48.55%yield) as a white solid. The crude product was used for next step directly without purification.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 7.69 -7.76 (m, 1 H) , 7.61 (d, J=7.70 Hz, 1 H) , 7.34 -7.51 (m, 3 H) , 7.15 -7.23 (m, 1 H) , 6.73 -6.94 (m, 2 H) , 4.49 -4.64 (m, 2 H) , 4.10 -4.20 (m, 1 H) , 1.42 (s, 9 H)

LC-MS: 348.2

General procedure for preparation of compound E-6:

To a solution of compound E-5 (0.70 g, 2.02 mmol, 1.00 eq) and DIEA (781 mg, 6.05 mmol, 1.05 mL, 3.00 eq) in DMF (30 mL) was added HATU (1.15 g, 3.02 mmol, 1.50 eq) . The reaction mixture was stirred at 20 ℃ for 16 hrs. The reaction mixture was poured into water (100 mL) . The aqueous phase was extracted with ethyl acetate (100 mL*3) . The combined organic phase was washed with brine (100 mL*2) , dried with anhydrous Na ₂SO ₄, filtered and concentrated in vacuum. The residue was purified by flash silica gel chromatography (

40 g

Silica Flash Column, Eluent of 0～50%THF/Petroleum ether gradient @40 mL/min) to afford compound E-6 (400 mg, 728 umol, 36.2%yield, 60%purity) as a white solid.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 8.11 (s, 1 H) , 7.83 -7.99 (m, 2 H) , 7.69 (t, J=7.15 Hz, 1 H) , 7.50 -7.60 (m, 1 H) , 7.29 (d, J=7.78 Hz, 1 H) , 6.88 (s, 1 H) , 6.66 (s, 1 H) , 4.41 -4.54 (m, 2 H) , 4.33 -4.38 (m, 1 H) , 1.36 (s, 9 H)

General procedure for preparation of compound E-7:

To a reaction mixture of compound E-6 (0.40 g, 728 umol, 60%purity, 1.00 eq) and K ₂CO ₃ (201 mg, 1.46 mmol, 2.00 eq) in DMF (5 mL) was added MeI (207 mg, 1.46 mmol, 90.7 uL, 2.00 eq) . The reaction mixture was stirred at 20 ℃ for 18 hrs. And then he reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with ethyl acetate (20 mL*3) . The combined organic phase was washed with brine (20 mL*2) , dried with anhydrous Na ₂SO ₄, filtered and concentrated in vacuum. The residue was purified by flash silica gel chromatography (

12 g

Silica Flash Column, Eluent of 0～50%THF/Petroleum ether gradient @30 mL/min) to afford compound E-7 (150 mg, 437 umol, 60.0%yield) as a white solid.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 8.19 (s, 1 H) , 7.96 (t, J=8.28 Hz, 2 H) , 7.72 -7.79 (m, 1 H) , 7.56 -7.64 (m, 1 H) , 7.36 (br d, J=8.03 Hz, 1 H) , 4.33 -4.56 (m, 3 H) , 3.46 (s, 3 H) , 1.32 -1.37 (m, 9 H) .

General procedure for preparation of compound E-8:

To a reaction mixture of compound E-7 (150 mg, 437 umol, 1.00 eq) in dioxane (2 mL) was added HCl/dioxane (4 M, 2.31 mL, 21.1 eq) . The reaction mixture was stirred at 20 ℃ for 4 hrs. And then the reaction mixture was concentrated under reduced pressure at 20 ℃. The residue was washed with TMBE: THF (10: 1, 10 mL) to afford desired product E-8 (100 mg, 357 umol, 81.8%yield, HCl salt) as a yellow solid. The crude product was used for next step directly without purification.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 8.60 (br s, 3 H) , 8.30 (s, 1 H) , 7.95 -8.02 (m, 2 H) , 7.71 -7.82 (m, 2 H) , 7.60 -7.65 (m, 1 H) , 4.49 -4.68 (m, 3 H) , 3.54 (s, 3 H)

LC-MS: 244.0

General procedure for preparation of compound 133:

To a solution of E-8 (50.0 mg, 178 umol, 1.00 eq, HCl) , 5-benzyl-4H-1, 2, 4-triazole-3-carboxylic acid (54.5 mg, 268 umol, 1.50 eq) and DIEA (69.3 mg, 536.2 umol, 93.4 uL, 3.00 eq) in DMF (1.5 mL) was added HATU (102 mg, 268 umol, 1.50 eq) . The reaction was stirred at 20 ℃ for 4 hrs. The residue was poured into water (10 mL) . The aqueous phase was extracted with ethyl acetate (10 mL*3) . The combined organic phase was washed with brine (5 mL*2) , dried with anhydrous Na ₂SO ₄, filtered and concentrated in vacuum. The crude product was purified by flash silica gel chromatography (

12 g

Silica Flash Column, Eluent of 0～50%THF/Petroleum ether gradient @30 mL/min) to afford desired crude product which was further purified by prep-HPLC (column: Phenomenex Gemini C18 250*50mm*10um; mobile phase: [water (0.225%FA) -ACN] ; B%: 35%-55%, 9min) to afford desired product compound 133 (16.8 mg, 37.64 umol, 21.06%yield, 96%purity) as a yellow solid.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 8.67 (s, 1 H) , 8.24 (s, 1 H) , 7.98 (t, J=7.91 Hz, 2 H) , 7.76 (t, J=7.78 Hz, 1 H) , 7.61 (t, J=8.03 Hz, 1 H) , 7.21 -7.37 (m, 5 H) , 4.93 -5.04 (m, 1 H) , 4.76 (br s, 1 H) , 4.53 (dd, J=9.66, 7.65 Hz, 1 H) , 4.13 (br s, 2 H) , 3.49 (s, 3 H) , 2.08 (s, 1 H)

LC-MS: 429.

Scheme IX below describes an illustrative synthetic scheme for preparation of compound 132 disclosed herein

Scheme IX: Synthesis scheme for compound 132

General procedure for preparation of compound E-10

To a solution of compound E-9 (20 g, 100 mmol, 1 eq) in MeCN (100 mL) was added TMSBr (76.9 g, 502 mmol, 65.2 mL, 5 eq) . The mixture was stirred at 90 ℃ for 16 h before it was cooled to 25 ℃. Filtered and the filter cake was dried under reduced pressure to give compound E-10 (30 g, crude) as a yellow solid which was used to next step without further purification.

¹H NMR (400MHz, DMSO-d6) δ ppm 8.15 -8.04 (m, 2H) , 7.95 (dd, J = 3.4, 6.4 Hz, 2H)

General procedure for preparation of compound E-11

To a solution of compound E-10 (15.0 g, 52.1 mmol, 1 eq) in DMSO (60 mL) MeCN (90 mL) was added K ₂CO ₃ (8.64 g, 62.5 mmol, 1.2 eq) and NH ₃. H ₂O (18.3 g, 156 mmol, 20.1 mL, 30%purity, 3 eq) . The mixture was stirred at 70 ℃for 15 h. The reaction mixture was quenched by addition water (300 mL) at 0 ℃, and then extracted with ethyl acetate (300 mL *2) . The combined organic layers were washed with water (300 mL) , dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

120 g

Silica Flash Column, Eluent of 22%Ethyl acetate/Petroleum ether gradient @85 mL/min) to afford compound E-11 (6.5 g, 27.6 mmol, 52.9%yield, 95.0%purity) as a light yellow solid.

General procedure for preparation of compound E-12

To a solution of compound E-46 (0.25 M, 64.2 mL, 1.2 eq) in THF (10 mL) was added K ₂CO ₃ (11.1 g, 80.3 mmol, 6 eq) in H ₂O (30 mL) . After stirring 25 ℃for 0.5 h, compound E-11 (3 g, 13.4 mmol, 1 eq) , Pd (PPh ₃) ₂Cl ₂ (470 mg, 670 umol, 0.05 eq) was added. The mixture was stirred at 70 ℃ for 1.5 h under N ₂. The reaction mixture was partitioned between water (100 mL) and ethyl acetate (200 mL) , the combine organic layers was dried over MgSO ₄, concentrated under reduce pressure to give a solid residue. The residue was purified by flash silica gel chromatography (

40 g

Silica Flash Column, Eluent of 50%Ethyl acetate/Petroleum ether gradient @60 mL/min) to afford compound E-12 (2.4 g, 4.26 mmol, 31.8%yield, 70%purity) as a yellow solid.

¹H NMR (400MHz, DMSO-d6) δ ppm 7.89 (d, J = 7.6 Hz, 1H) , 7.72 (d, J = 8.1 Hz, 1H) , 7.52 -7.48 (m, 2H) , 7.42 -7.34 (m, 5H) , 6.79 (s, 2H) , 5.06 (s, 2H) , 4.30 -4.25 (m, 2H) , 3.66 (s, 3H) , 3.71 -3.61 (m, 1H) , 2.94 -2.79 (m, 2H) , 2.28 -2.08 (m, 2H) .

General procedure for preparation of compound E-13

To a solution of compound E-12 (5 g, 12.7 mmol, 1 eq) in MeOH (40 mL) , H ₂O (40 mL) was added LiOH. H ₂O (797 mg, 19.0 mmol, 1.5 eq) . The mixture was stirred at 25 ℃ for 1 h. The aqueous phase was acidified by HCl (1 N) until PH = 7, and then extracted by ethyl acetate (50 mL *3) . The combined layers were dried over MgSO ₄, filtered and concentrated under reduced pressure to afford compound E-13 (3 g, 6.70 mmol, 52.8%yield, crude) as a yellow solid which was used to next step without further purification.

General procedure for preparation of compound E-14

To a solution of compound E-13 (3 g, 7.89 mmol, 1 eq) in DCM (30 mL) DMF (30 mL) was added N, N'-dicyclohexylmethanediimine (1.95 g, 9.46 mmol, 1.91 mL, 1.2 eq) , 1-hydroxybenzotriazole (1.28 g, 9.46 mmol, 1.2 eq) and 4-methylmorpholine (797 mg, 7.89 mmol, 867 uL, 1 eq) . The mixture was stirred at 25 ℃ for 10 h. The reaction mixture was diluted with water (100 mL) and extracted with DCM (100 mL *3) . The combined organic layers were dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

25 g

Silica Flash Column, Eluent of 50%THF/Petroleum ether gradient @40 mL/min) . Compound E-14 (2.5 g, 6.21 mmol, 78.7%yield, 90%purity) was obtained as a yellow solid.

¹H NMR (400MHz, DMSO-d6) δ ppm 10.91 (s, 1H) , 8.03 (dd, J = 1.3, 8.1 Hz, 1 H) , 7.94 (dd, J = 1.2, 8.2 Hz, 1H) , 7.83 -7.72 (m, 3H) , 7.39 -7.27 (m, 5H) , 5.04 -4.95 (m, 2H) , 4.18 -4.09 (m, 1H) , 3.15 -3.06 (m, 2H) , 2.47 -2.37 (m, 1H) , 2.30 -2.18 (m, 1H)

General procedure for preparation of compound E-15

To a solution of compound E-14 (0.5 g, 1.38 mmol, 1 eq) , MeI (587 mg, 4.14 mmol, 257 uL, 3 eq) in DMF (1 mL) was added K ₂CO ₃ (572 mg, 4.14 mmol, 3 eq) . The mixture was stirred at 25 ℃ for 5 h. and then was quenched with water (50 mL) and extracted with DCM (50 mL *2) . The combined organic layers were dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

4 g

Silica Flash Column, Eluent of 50%THF/Petroleum ether gradient @10 mL/min) . Compound E-15 (0.4 g, 956 umol, 69.3%yield, 90%purity) was obtained as a yellow solid.

¹H NMR (400MHz, DMSO-d6) δ ppm 8.11 -8.00 (m, 2H) , 7.87 -7.77 (m, 3H) , 7.45 -7.21 (m, 5H) , 5.07 -4.92 (m, 2H) , 4.12 (td, J = 7.6, 12.0 Hz, 1H) , 3.48 (s, 3H) , 3.23 -2.93 (m, 2H) , 2.40 (s, 1H) , 2.27 -2.12 (m, 1H)

General procedure for preparation of compound E-16

To a solution of compound E-15 (100 mg, 276 umol, 1 eq) in CH ₃COOH (1 mL) was added HBr (1.49 g, 5.52 mmol, 998 uL, 30%purity, 20 eq) . The mixture was stirred at 25 ℃ for 1 h and then was concentrated under reduced pressure to give compound E-16 (0.85 g, crude, HBr salt) as a yellow solid which was used to next step without further purification.

¹H NMR (400MHz, DMSO-d6) δ ppm 8.36 (br d, J = 3.5 Hz, 3H) , 8.06 (dd, J = 1.5, 7.9 Hz, 1H) , 7.99 -7.90 (m, 1H) , 7.81 (dt, J = 1.2, 8.3 Hz, 2H) , 4.39 -4.21 (m, 1H) , 4.39 -4.21 (m, 1H) , 3.27 -3.07 (m, 2H) , 2.71 -2.57 (m, 1H) , 2.34 -2.22 (m, 1H) , 1.91 (s, 3H) .

General procedure for preparation of compound 132:

To a solution of 5-benzyl-4H-1, 2, 4-triazole-3-carboxylic acid (241 mg, 1.19 mmol, 3 eq) in Py (3 mL) was added EDCI (228 mg, 1.19 mmol, 3 eq) . Then compound E-16 (128 mg, 396 umol, 1 eq, HBr) was added. The mixture was stirred at 20 ℃ for 16 h. The reaction concentrated under reduced pressure to give a residue. The residue was partitioned between EtOAc (20 mL) and water (15 mL) . The organic phase was separated, concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: YMC-Actus Triart C18 150*30mm*5um; mobile phase: [water (0.05%ammonia hydroxide v/v) -ACN] ; B%: 30%-50%, 10min) to afford compound 132 (50.8 mg, 119 umol, 30.0%yield, 100%purity) as a white solid.

¹H NMR: (400MHz, DMSO-d ₆) δ ppm 8.64 (br s, 1H) , 8.10 -8.05 (m, 1H) , 8.01 (d, J=7.5 Hz, 1H) , 7.88 -7.75 (m, 2H) , 7.38 -7.29 (m, 2H) , 7.28 -7.20 (m, 3H) , 4.57 -4.44 (m, 1H) , 4.10 (s, 2H) , 3.48 (s, 3H) , 3.45 -3.40 (m, 1H) , 3.30 (br s, 1H) , 3.25 -3.13 (m, 1H) , 3.12 -3.03 (m, 1H)

LCMS: 428.1

Scheme X below describes an illustrative synthetic scheme for preparation of compound 106 disclosed herein.

Scheme X: Synthesis scheme for compound 106

General procedure for preparation of compound E-18

To a stirred suspension of compound E-17 (200 g, 1.59 mol, 1 eq) in Py (1000 mL) was added TosCl (604 g, 3.17 mol, 2 eq) in portions at 25 ℃. The mixture was stirred at 85 ℃ for 16 h. The reaction mixture was concentrated under reduced pressure to remove most of solvent, diluted with HCl (3 N) 6 L and filtered. The filter cake was triturated with ethanol (1500 mL) , filtered and dried under reduced pressure to give compound E-18 (666 g, 1.38 mol, 87.0%yield, 90%purity) as a red solid which was used to next step without further purification. LCMS: 457.0

¹HNMR ES14701-78-P1_NMR (400MHz, DMSO-d ₆) δ 9.55 -9.30 (m, 2H) , 9.55 -9.30 (m, 1H) , 7.67 (d, J = 8.3 Hz, 2H) , 7.54 (d, J = 8.3 Hz, 2H) , 7.37 (t, J = 8.8 Hz, 4H) , 6.92 -6.77 (m, 3H) , 2.37 (s, 6H)

General procedure for preparation of compound E-19:

To a stirred suspension of compound E-18 (50.0 g, 115 mmol, 1 eq) in AcOH (500 mL) was added a solution of HNO ₃ (12.4 g, 138 mmol, 8.88 mL, 70%purity, 1.2 eq) in AcOH (50 mL) at 25 ℃. The mixture was stirred at 65 ℃ for 0.5 h and then was cooled to 25 ℃, filtered. The filter cake was washed with ethanol 20 mL, filtered and dried under reduced pressure to give compound E-19 (420 g, 0.8 mol, 72.5%yield) as a white solid which was used to next step without further purification.

LCMS: 502.0

¹H NMR (400MHz, DMSO-d ₆) δ 7.78 (d, J = 8.3 Hz, 2H) , 7.62 -7.56 (m, 3H) , 7.39 (dd, J = 8.3, 11.3 Hz, 4H) , 7.19 (d, J = 13.6 Hz, 1H) , 2.37 (s, 6H)

General procedure for preparation of compound E-20

A suspension of compound E-19 (151 g, 315 mmol, 1 eq) in H ₂SO ₄ (600 mL, 90%purity) was stirred at 85 ℃ for 0.5 h and then was cooled to 25 ℃, added to ice water (4000 mL) dropwise, alkalized by NH ₃. H ₂O till pH=9. A lot of solid was formed, filtered. The filter cake was dried under reduced pressure to afford compound E-20 (61 g, crude) as a yellow solid, which was used to next step without further purification.

LCMS: 172.1

¹H NMR (400MHz, DMSO-d ₆) δ = 7.24 (d, J = 7.8 Hz, 1H) , 6.41 -6.33 (m, 3H) , 4.92 (br s, 2H)

General procedure for preparation of compound E-21

To a stirred solution of compound E-20 (80 g, 467 mmol, 1 eq) in ethanol (800 mL) was added oxaldehyde (81.4 g, 1.40 mol, 73.3 mL, 3 eq) in one potion at 25 ℃. The mixture was heated to 50 ℃ and stirred for 0.5 h and then was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

330 g

Silica Flash Column, Eluent of 0-30%Ethyl acetate/Petroleum ether gradient @100 mL/min) to give compound E-21 (76 g, 393.51 mmol, 84.2%yield) as a yellow solid.

LCMS: 242.2

¹H NMR (400MHz, DMSO-d ₆) δ = 9.15 (dd, J = 1.7, 9.4 Hz, 2H) , 8.93 (d, J = 7.8 Hz, 1H) , 8.32 (d, J = 11.6 Hz, 1H)

General procedure for preparation of compound E-22

To a stirred solution of Boc-L-serine (88.0 g, 429 mmol, 1.2 eq) in DMF (400 mL) was added NaH (34.3 g, 857 mmol, 60%purity, 2.4 eq) in portions at 0 ℃ under N ₂ over 0.5 h. The mixture was stirred at 0 ℃ for 0.5 h till no more H ₂ released. Then to the mixture was added a suspension of compound E-21 (69.0 g, 357 mmol, 1 eq) in DMF (400 mL) at 0 ℃ dropwise over 0.5 h. After the addition, the mixture was allowed to warm to 20 ℃ over a period of 2 h under N ₂ to give a dark brown suspension. The reaction mixture was cooled to 0 ℃, poured into ice water (1500 mL) slowly, acidified with 2 N HCl to pH=3, extracted with Ethyl Acetate (1500 mL *2) . The combined organic layers were washed with brine (1500 mL) , dried over Na ₂SO ₄, filtered and concentrated under reduced pressure to give compound E-22 (144 g, crude) as a brown oil, which was used to next step without further purification.

LCMS: 559.1

¹H NMR (400MHz, DMSO-d ₆) δ = 9.04 (d, J = 1.8 Hz, 1H) , 8.96 (d, J = 1.7 Hz, 1H) , 8.65 (s, 1H) , 7.85 (s, 1H) , 4.61 -4.52 (m, 2H) , 3.99 -3.94 (m, 1H) , 1.38 (s, 9H)

General procedure for preparation of compound E-23

To a suspension of compound E-22 (144 g, 380.61 mmol, 1 eq) in ethyl acetate (1000 mL) was added Pd/C (16 g, 10%purity) under N ₂ at 25 ℃. The suspension was degassed under vacuum and purged with H ₂ 3 times. The mixture was stirred under H ₂ (15psi) at 25℃ for 15 h. The reaction mixture was filtered and concentrated under reduced pressure to give compound E-23 as a brown oil.

¹H NMR (400MHz, DMSO-d ₆) δ = 8.50 (d, J = 2.0 Hz, 1H) , 8.43 (d, J = 2.0 Hz, 1H) , 7.22 (s, 1H) , 6.97 (s, 1H) , 6.14 -5.96 (m, 2H) , 4.64 -4.56 (m, 2H) , 4.00 -3.93 (m, 1H) , 1.39 (s, 9H)

General procedure for preparation of compound E-24

To a suspension of compound E-23 (36.0 g, 103 mmol, 1 eq, crude) in ethyl acetate (100 mL) was added T ₃P (98.7 g, 155 mmol, 92.2 mL, 50%purity, 1.5 eq) and DIEA (53.4 g, 413 mmol, 72.0 mL, 4 eq) . The mixture was stirred at 50 ℃ for 1 h. The reaction mixture was cooled to 25 ℃, diluted with saturated NaHCO ₃ (100 mL) and extracted with ethyl acetate (100 mL *2) . The combined organic layers were washed with brine (200 mL) , dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

120 g

Silica Flash Column, Eluent of 0%～66%Ethyl acetate/Petroleum ether gradient @30 mL/min) . Compound E-24 (30 g, 90.0 mmol, 95%purity) was obtained as a light yellow solid.

LCMS: 331.2

¹H NMR (400MHz, DMSO-d ₆) δ = 10.51 (s, 1H) , 8.86 (d, J = 3.9 Hz, 2H) , 7.78 (s, 1H) , 7.73 (s, 1H) , 7.25 (d, J = 7.6 Hz, 1H) , 4.52 -4.36 (m, 3H) , 1.37 (s, 9H)

General procedure for preparation of compound E-25

To a solution of compound E-24 (16 g, 45.4 mmol, 1 eq) in DMF (120 mL) was added K ₂CO ₃ (12.6 g, 90.6 mmol, 2 eq) and MeI (10.3 g, 67.1 mmol, 4.23 mL, 1.5 eq) . The mixture was stirred at 25 ℃ for 3 h. The reaction mixture was partitioned between water 600 mL and ethyl acetate 1000 mL (500mL *2) . The combined organic layers were washed with brine (600 mL) , dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

80 g

Silica Flash Column, Eluent of 65%Ethyl acetate/Petroleum ether gradient @100 mL/min) . Compound E-25 (15.2 g, 44.1 mmol, 91.1%yield) was obtained as a white solid.

LCMS: 344.9

¹H NMR (400MHz, DMSO-d ₆) δ = 8.93 (d, J = 6.8 Hz, 2H) , 8.18 (s, 1H) , 7.85 (s, 1H) , 7.30 (d, J = 8.3 Hz, 1H) , 4.53 -4.35 (m, 3H) , 3.47 (s, 3H) , 1.34 (s, 9H)

General procedure for preparation of compound E-26

To a solution of compound E-25 (14 g, 40.6 mmol, 1 eq) in DMF (130 mL) was added NCS (5.97 g, 44.7 mmol, 1.1 eq) at 25 ℃. The mixture was stirred at 80 ℃ for 3 h. The reaction mixture was cooled to 25 ℃, partitioned between water 1000 mL and DCM 2000 mL (1000 mL *2) . The combined organic layers were washed with brine (1000 mL) , dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

120 g

Silica Flash Column, Eluent of 28%Ethyl acetate/Petroleum ether gradient @60 mL/min) . Compound E-26 (8.2 g, 21.4 mmol, 52.7%yield, 99%purity) was obtained as a light yellow solid.

LCMS: 379.1

¹H NMR (400MHz, CDCl ₃) δ = 8.89 (d, J = 1.6 Hz, 1H) , 8.86 (d, J = 1.6 Hz, 1H) , 7.77 (s, 1H) , 5.49 (br d, J = 6.5 Hz, 1H) , 4.64 -4.44 (m, 2H) , 4.18 -4.02 (m, 1H) , 3.43 (s, 3H) , 1.32 (s, 9H)

General procedure for preparation of compound E-27

To a solution of compound E-26 (9.00 g, 23.8 mmol, 1 eq) in dioxane (30 mL) was added HCl/dioxane (4 M, 47.5 mL, 8 eq) at 25 ℃. The mixture was stirred at 25 ℃ for 5 h. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was used to next step without further purification. Compound E-27 (7.5 g, 22.6 mmol, 95.2%yield, 95%purity, HCl salt) was obtained as a yellow solid.

LCMS: 278.9

¹H NMR (400MHz, DMSO-d ₆) δ = 9.10 (s, 2H) , 8.77 (br s, 3H) , 8.03 (s, 1H) , 4.65 -4.40 (m, 3H) , 3.41 (s, 3H)

General procedure for preparation of compound 106

To a stirred solution of 5-benzyl-4H-1, 2, 4-triazole-3-carboxylic acid (8.5 g, 32.4 mmol, 80%purity, 1.2 eq) in DMF (85 mL) was added HOBt (5.47 g, 40.5 mmol, 1.5 eq) , EDCI (7.76 g, 40.5 mmol, 1.5 eq) and DIPEA (17.4 g, 135 mmol, 23.5 mL, 5 eq) . The mixture was stirred at 25 ℃ for 1 h. And then to the mixture was added compound E-27 (8.5 g, 26.9 mmol, 1 eq, HCl salt) , stirred at 25 ℃ for 9 h. The reaction mixture was partitioned between water 500 mL and DCM 1500 mL (500 mL *3) . The organic phase was separated, washed with saturated solution of NaCl in water, dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

220 g

Silica Flash Column, Eluent of 53%THF/n-hexane gradient @100mL/min) . And then slurred in TBME and THF (9: 1) 500 (mL) , lyophilized to afford compound 106 (7.2 g, 15.4 mmol, 57.0%yield, 99%purity) as a white solid.

LCMS: 464.1

¹H NMR (400 MHz, CDCl ₃) δ = 11.90 (br s, 1H) , 8.98 (dd, J = 1.6, 7.9 Hz, 2H) , 8.13 (d, J = 7.3 Hz, 1H) , 7.90 (s, 1H) , 7.30 (br s, 2H) , 7.27 (br s, 3H) , 5.20 -4.93 (m, 1H) , 4.70 (dd, J = 7.7, 9.7 Hz, 1H) , 4.32 (t, J = 10.5 Hz, 1H) , 4.18 (s, 2H) , 3.54 (s, 3H)

Scheme XI below describes an illustrative synthetic scheme for preparation of compound 109 disclosed herein.

Scheme XI: Synthesis scheme for compound 109

General procedure for preparation of compound E-29:

To a solution of compound E-28 (50.0 g, 267 mmol, 1 eq) in pyridine (500 mL) was added TosCl (102 g, 534 mmol, 2 eq) in portions at 0 ℃. After the addition, the mixture was stirred at 85 ℃ for 18 hrs. The mixture was concentrated to remove most of the solvent. The residue was poured into a stirred 1 M of HCl (3 L) . Solid precipitated. The mixture was stirred at 25 C for 30 min, filtered and the filter cake was washed with water (300 mL) , EtOH (100 mL) , dried under vacuum to give compound E-29 (120 g, 242 mmol, 90.6%yield) as a yellow solid, which was directly used in the next step without further purification.

¹H NMR: (400 MHz, DMSO-d ₆) δ ppm 9.26 -9.67 (m, 2 H) , 7.60 (t, J=7.69 Hz, 4 H) , 7.38 (t, J=8.44 Hz, 4 H) , 7.20 (dd, J=8.63, 2.25 Hz, 1 H) , 7.13 (d, J=2.25 Hz, 1 H) , 6.93 (d, J=8.76 Hz, 1 H) , 2.37 (d, J=2.75 Hz, 6 H)

General procedure for preparation of compound E-30

A prepared solution of nitric acid (12.0 g, 133 mmol, 8.57 mL, 70%purity, 1.1 eq) in AcOH (25 mL) was added dropwise to a suspension of compound E-29 (60.0 g, 121 mmol, 1 eq) in AcOH (500 mL) at 65 ℃ within a period of 30 min. After the addition, the mixture was maintained at the same temperature for a further 60 min. The mixture turned yellow. TLC (petroleum ether/EtOAc=1/1, UV) showed the reaction was completed. The mixture was cooled to 20 ℃, filtered and the filter cake was dried under vacuum to give compound E-30 (91.0 g, 1689 mmol, 69.5%yield) as a light yellow solid, which was directly used in the next step without further purification.

¹H NMR: (400 MHz, DMSO-d ₆) δ ppm 7.73 (s, 1 H) , 7.59 -7.70 (m, 3 H) , 7.59 -7.70 (m, 1 H) , 7.43 -7.47 (m, 1 H) , 7.39 (t, J=7.03 Hz, 4 H) , 2.37 (s, 6 H)

General procedure for preparation of compound E-31:

To H ₂O (18 mL) was added H ₂SO ₄ (180 mL, 98%purity) dropwise at 0 ℃. Then compound E-30 (50 g, 92.52 mmol, 1 eq) was added in portions. After the addition, the mixture was stirred at 85 ℃ for 0.5 h. The suspension turned yellow clear. LCMS showed the reaction was completed. The mixture was poured into ice water (1000 mL) , basified withe NH ₃. H ₂O to pH～10. Red solid precipitated. The solid was collected by filtration, dried under vacuum to give compound E-31 (19.6 g, 84.5 mmol, 91.3%yield) as a red solid, which was directly used in the next step without further purification.

LC-MS: 233.8

General procedure for preparation of compound E-32

To a solution of compound E-31 (19.0 g, 81.9 mmol, 1 eq) in EtOH (200 mL) and H ₂O (10 mL) was added oxaldehyde (14.2 g, 245 mmol, 12.8 mL, 3 eq) . The mixture was stirred at 90 ℃ for 0.5 hr. The mixture turned light yellow and solid precipitated. The mixture was filtered and the filter cake was dried under vacuum to give compound E-32 (19.6 g, 77.1 mmol, 94.2%yield) as a yellow solid, which was directly used in the next step without further purification.

¹H NMR: (400 MHz, DMSO-d ₆) δ ppm 9.15 (s, 2 H) , 8.85 (s, 1 H) , 8.70 (s, 1 H)

General procedure for preparation of compound E-33

To a suspension of compound E-32 (11.0 g, 43.3 mmol, 1 eq) in EtOH (200 mL) and H ₂O (40 mL) was added NH ₄Cl (4.63 g, 86.6 mmol, 2 eq) and Fe (9.67 g, 173 mmol, 4 eq) . The mixture was stirred at 80 ℃ for 1 hr. The mixture turned brown. The mixture was filtered through a pad of celite and the filtrate was concentrated to give compound E-33 (9.10 g, crude) as a brown solid.

LCMS: 225.9

General procedure for preparation of compound E-34:

To a suspension of compound E-33 (7.20 g, 32.1 mmol, 1 eq) in THF (30 mL) and H ₂O (60 mL) was added K ₂CO ₃ (26.6 g, 193 mmol, 6 eq) and compound E-46. The mixture was degassed with N ₂ for 2 minutes. Then Pd-118 (1.05 g, 1.61 mmol, 0.05 eq) was added under N ₂. The resulting mixture was stirred at 60 ℃ for 2 hrs. The cooled mixture was then diluted with EtOAc (200 mL) , washed with brine (200 mL*2) , dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give compound E-34 (12.0 g, crude) as a brown gum.

LCMS: 395.1

General procedure for preparation of compound E-35

To a solution of compound E-34 (11.0 g, 27.9 mmol, 1 eq) in THF (150 mL) and H ₂O (40 mL) was added LiOH. H ₂O (4.68 g, 111 mmol, 4 eq) . The mixture was stirred at 50 ℃ for 1 hr. The mixture was diluted with H ₂O (20 mL) , extracted with EtOAc (200 mL) . The EtOAc layer was discarded. The aqueous layer was acidified with 2 N HCl to pH～4, solid precipitated. The solid was collected by filtration, dried under vacuum to give compound E-35 (8.2 g, 21.5 mmol, 77.3%yield) as a yellow solid.

LCMS: 381.1

General procedure for preparation of compound E-36:

To a solution of compound E-35 (8.00 g, 21.0 mmol, 1 eq) in EtOAc (300 mL) was added T ₃P (26.7 g, 42.06 mmol, 25.0 mL, 50%purity, 2 eq) and TEA (8.51 g, 84.1 mmol, 11.7 mL, 4 eq) . The mixture was stirred at 50 ℃ for 20 min. The mixture was yellow clear. LCMS showed the reaction was completed. The mixture was diluted with EtOAc (200 mL) , washed with sat. NaHCO ₃ (300 mL) , brine (300 mL) , dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give a yellow solid. This solid was purified by flash silica gel chromatography (

40 g

Silica Flash Column, Eluent of 0～4%MeOH/DCM gradient @40 mL/min) to give compound E-36 (5.80 g, 16.0 mmol, 76.1%yield) as a yellow solid.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 8.74 -8.88 (m, 2 H) , 8.44 (br s, 1 H) , 7.98 (s, 1 H) , 7.73 (s, 1 H) , 7.27 -7.37 (m, 5 H) , 5.83 (br d, J=7.28 Hz, 1 H) , 5.06 (s, 2 H) , 4.34 -4.41 (m, 1 H) , 3.53 (s, 1 H) , 3.05 -3.17 (m, 1 H) , 2.96 -3.03 (m, 1 H) , 2.67 -2.83 (m, 1 H) .

General procedure for preparation of compound E-37

To a solution of compound E-36 (1.02 g, 2.81 mmol, 1 eq) in DMF (10 mL) was added K ₂CO ₃ (778 mg, 5.63 mmol, 2 eq) and MeI (479 mg, 3.38 mmol, 210 uL, 1.2 eq) . The mixture was stirred at 15 ℃ for 16 hrs. The mixture was diluted with EtOAc (50 mL) , washed with brine (30 mL*3) . The EtOAc layer was dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give compound E-37 (0.96 g, 2.55 mmol, 90.6%yield) as a yellow solid.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 8.84 (q, J=1.79 Hz, 2 H) , 7.97 (s, 1 H) , 7.90 (s, 1 H) , 7.27 -7.37 (m, 5 H) , 5.80 (br d, J=7.46 Hz, 1 H) , 5.03 (s, 2 H) , 4.24 -4.40 (m, 2 H) , 3.56 (s, 3 H) , 2.91 -3.08 (m, 2 H) , 2.66 (tt, J=12.53, 7.76 Hz, 1 H)

General procedure for preparation of compound E-38

To a solution of compound E-37 (50 mg, 133 umol, 1 eq) in AcOH (0.6 mL) was added HBr (2.24 g, 9.12 mmol, 1.5 mL, 33%purity, 68.6 eq) under N ₂. The mixture rapidly turned black. The mixture was stirred at 20 ℃ 2 hrs. The mixture was concentrated to give compound E-38 (42.9 mg, crude, HBr salt) as a brown solid.

LC-MS: 243.0

General procedure for preparation of compound 109:

To a mixture of 1-oxidopyridin-1-ium-2-ol (39.4 mg, 355 umol, 2 eq) , EDCI (68.1 mg, 355 umol, 2 eq) and TEA (71.8 mg, 710 umol, 98.8 uL, 4 eq) in DMF (3 mL) was added compound E-38 (42.9 mg, 177 umol, 1 eq, HBr salt) and 5-benzyl- 4H-1, 2, 4-triazole-3-carboxylic acid (72.1 mg, 355 umol, 2 eq) . The mixture was stirred at 50 ℃ for 1 hr. The mixture was concentrated, and the residue was partitioned between H ₂O (10 mL) and DCM (10 mL) . The DCM layer was dried over Na ₂SO ₄, filtered and the filtrate was concentrated to give yellow residue. The residue was purified by prep-HPLC (column: YMC Triart C18 250*50mm*7um; mobile phase: [water (0.05%ammonia hydroxide v/v) -ACN] ; B%: 3%-43%, 12min) , lyophilized under high vacuum to give a yellow solid (32 mg) . This solid was dissolved in EtOAc (20 mL) , washed with DI water (20 mL*3) , dried under vacuum and lyophilized under high vacuum to give compound 109 (18.1 mg, 41.8 umol, 23.6%yield, 98.7%purity) as a light yellow solid.

LC-MS: 428.2

¹H NMR: (400 MHz, DMSO-d6) δ ppm 14.41 (br s, 1 H) , 14.08 -14.70 (m, 1 H) , 8.85 -8.99 (m, 2 H) , 8.40 (br s, 1 H) , 8.10 (d, J=7.78 Hz, 2 H) , 7.19 -7.34 (m, 5 H) , 4.38 (dt, J=11.48, 7.69 Hz, 1 H) , 4.09 (s, 2 H) , 3.47 (s, 3 H) , 3.06 (dd, J=12.80, 7.03 Hz, 1 H) , 2.90 (td, J=12.61, 8.66 Hz, 1 H) , 2.20 -2.45 (m, 2 H)

Scheme XII below describes an illustrative synthetic scheme for preparation of compound 131 disclosed herein.

Scheme XII: Synthesis scheme for compound 131

General procedure for preparation of compound E-40:

To a solution of compound E-39 (26 g, 63.1 mmol, 1 eq) in ethanol (200 mL) and water (200 mL) was added NH ₄Cl (27.0 g, 505 mmol, 8 eq) and Fe (24.7 g, 442 mmol, 7 eq) . The mixture was stirred at 60 ℃ for 4 h. And then the reaction mixture was filtered and partitioned between water 500mL and ethyl acetate (1000 mL) . The organic phase was separated, washed with brine (500 mL) , dried over MgSO ₄, filtered and concentrated under reduced pressure to give compound E-40 (18 g, crude) as a yellow solid which was used to next step without further purification.

General procedure for preparation of compound E-41

To a solution of compound E-40 (18.0 g, 47.1 mmol, 1 eq) in MeOH (150 mL) , H ₂O (30 mL) was added K ₂CO ₃ (13.0 g, 94.2 mmol, 2 eq) . The mixture was stirred at 25 ℃ for 10 h. And then the reaction mixture was concentrated under reduced pressure to give compound E-41 (14 g, crude) as a yellow solid which was used to next step without further purification.

General procedure for preparation of compound E-42

To a solution of compound E-41 (13.5 g, 47.1 mmol, 1 eq) in THF (200 mL) was added TEA (14.3 g, 141 mmol, 19.7 mL, 3 eq) and Boc ₂O (15.4 g, 70.7 mmol, 16.2 mL, 1.5 eq) . The mixture was stirred at 25 ℃ for 2 h. And then the reaction mixture was partitioned between ethyl acetate (1000 mL) and water (500 mL) . The organic phase was separated, dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

120 g

Silica Flash Column, Eluent of 30%Ethyl acetate/Petroleum ether gradient @30 mL/min) . Compound E-42 (13.5 g, 32.6 mmol, 69.3%yield, 90%purity) was obtained as a yellow solid.

LC-MS: 330.1

¹H NMR: (400MHz, CDCl ₃) δ ppm 7.25 (s, 1H) , 6.55 (s, 1H) , 5.50 (br d, J=6.8 Hz, 1H) , 4.73 -4.63 (m, 1H) , 4.52 (dd, J=7.2, 9.7 Hz, 1H) , 4.23 -4.08 (m, 3H) , 3.34 (s, 3H) , 1.42 (s, 9H)

General procedure for preparation of compound E-43

To a solution of compound E-42 (0.5 g, 1.29 mmol, 1 eq) in DMSO (2 mL) was added CuI (49.31 mg, 259 umol, 0.2 eq) and K ₃PO ₄ (549 mg, 2.59 mmol, 2 eq) N, N'-bis (4-methyl-2-phenyl-phenyl) oxamide (109 mg, 258.91 umol, 0.2 eq) NH ₃.H ₂O (544 mg, 3.88 mmol, 598 uL, 25%purity, 3 eq) . The mixture was stirred at 80 ℃ for 12 h. The reaction mixture was filtered, partitioned between water (30 mL) and ethyl acetate (30 mL *2) . The combined organic layers were dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

40 g

Silica Flash Column, Eluent of 40%THF/Petroleum ether gradient @20 mL/min) . And compound E-43 (0.7 g, 1.95 mmol, 25.2%yield, 90%purity) was obtained as a yellow solid.

LC-MS: 375.3

¹H NMR: (400MHz, CDCl ₃) δ ppm 6.51 (d, J=11.1 Hz, 2H) , 5.50 (br d, J=7.0 Hz, 1H) , 4.68 (td, J=7.5, 11.0 Hz, 1H) , 4.51 (dd, J=7.4, 9.6 Hz, 1H) , 4.13 -4.00 (m, 1H) , 3.57 -3.34 (m, 4H) , 3.33 (s, 3H) , 1.41 (s, 9H)

General procedure for preparation of compound E-44

To a solution of compound E-43 (200 mg, 620 umol, 1 eq) in EtOH (2 mL) was added butane-2, 3-dione (58.8 mg, 682 umol, 59.6 uL, 1.1 eq) . The mixture was stirred at 50 ℃ for 1 h. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (

4 g

Silica Flash Column, Eluent of 40% Ethyl acetate/Petroleum ether gradient @40 mL/min) . And compound E-44 (0.19 g, 484 umol, 78.1%yield, 95%purity) was obtained as a white solid.

¹H NMR (400MHz, CDCl ₃) δ ppm 7.80 (s, 1H) , 7.73 (s, 1H) , 5.55 (br d, J=7.0 Hz, 1H) , 4.76 -4.61 (m, 2H) , 4.23 (dd, J=9.6, 11.2 Hz, 1H) , 3.55 (s, 3H) , 2.75 (d, J=2.7 Hz, 6H) , 1.41 (s, 9H)

General procedure for preparation of compound E-45

To a solution of compound E-44 (130.00 mg, 349.07 umol, 1 eq) in dioxane (1 mL) was added HCl/dioxane (4 M, 872.68 uL, 10 eq) . The mixture was stirred at 25 ℃ for 1 h. The reaction mixture was concentrated under reduced pressure to give compound E-45 (0.107 g, 346 umol, 99.3%yield, HCl) as a yellow solid which was used to next step without further purification.

General procedure for preparation of compound 131:

To a solution of compound E-45 (0.107 g, 393 umol, 1 eq, HCl) 5-benzyl-4H-1, 2, 4-triazole-3-carboxylic acid (79.9 mg, 393 umol, 1 eq) in DMF (2 mL) was added DIEA (254 mg, 1.96 mmol, 342.2 uL, 5 eq) and T3P (500.11 mg, 786 umol, 467 uL, 50%purity, 2 eq) . The mixture was stirred at 50 ℃ for 1 h. And then the reaction mixture was partitioned between water (20 mL) and ethyl acetate (20 mL*2) . The organic phase was separated, washed with water (10 mL) , dried over MgSO ₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Gemini-NX 150*30mm*5um; mobile phase: [water (0.05%ammonia hydroxide v/v) -ACN] ; B%: 25%-45%, 11 min) to give desired compound 132 (0.08 g, 173 umol, 44.1%yield, 99%purity) as a white solid.

LCMS: 458.2

¹H NMR (400MHz, CDCl ₃) δ ppm 11.37 (br s, 1H) , 8.12 (br d, J=7.1 Hz, 1H) , 7.84 (s, 1H) , 7.78 (s, 1H) , 7.36 -7.30 (m, 2H) , 7.28 -7.23 (m, 3H) , 5.14 (td, J=7.3, 11.2 Hz, 1H) , 4.77 (dd, J=7.4, 9.7 Hz, 1H) , 4.34 (dd, J=10.0, 11.1 Hz, 1H) , 4.18 (s, 2H) , 3.58 (s, 3H) , 2.76 (d, J=4.6 Hz, 6H)

Compounds in Table 5 below were prepared by following procedures that are the same as or similar to those for the preparation of compounds 106, 109, and 131-133 as described above.

Table 5

Scheme XIV below describes an illustrative synthetic scheme for preparation of compound 144 disclosed herein.

Scheme XIV: Synthesis scheme for compound 144:

General procedure for preparation of compound G-2

Glycerol (72.0 g, 782 mmol, 58.5 mL, 2.70 eq) was heated to 160 ℃ and stirred at 160 ℃ for 0.5 h, then cooled to 110 ℃, G-1 (50.0 g, 289 mmol, 1.00 eq) and NaI (868 mg, 5.79 mmol, 0.02 eq) was added, the mixture stirred at 110 ℃ to get a homogeneous tar. The mixture was again heated to 150 ℃ and H ₂SO ₄ (56.8 g, 579 mmol, 30.8 mL, 2.00 eq) was added dropwise. The mixture was stirred at 150 ℃ for 18 hrs. The cooled mixture was poured into ice water (1000 mL) , basified with solid NaHCO ₃ till pH=8, extracted with DCM (500 mL*2) , the organic phase was concentrated in vacuum to give a yellow solid. The residue was purified by silica gel chromatography (

330 g

Silica Flash Column, Eluent of 0～10%Ethyl acetate/Petroleum ether gradient @100 mL/min) . And compound G-2 (15.0 g, 71.9 mmol, 24.8%yield) was obtained as a white solid.

LCMS: 208.9

General procedure for preparation of compound G-3

To a solution of Boc-L-serine (23.6 g, 115 mmol, 1.20 eq) in DMF (300 mL) was added NaH (9.20 g, 230 mmol, 60.0 %purity, 2.40 eq) under N ₂ at 0 ℃. The mixture was stirred at 0 ℃ for 2 hrs. Then G-2 (20.0 g, 95.8 mmol, 1.00 eq) was added at 0℃. The mixture was stirred at 25 ℃ for 16 hrs and then was poured into ice water (400 mL) . To the aqueous phase was added HCl (3 M) till pH=5-6, then extracted with EtOAc (350 mL*2) , the organic phase washed with brine (250 mL*3) , dried with Na ₂SO ₄, filter and concentrated in vacuum. The residue was purified by flash silica gel chromatography (

220 g

Silica Flash Column, Eluent of 0～74%Ethyl acetate/Petroleum ether gradient @ 60mL/min) . And compound G-3 (10.0 g, 26.5 mmol, 27.6%yield) was obtained as brown oil.

LCMS: 377.5

General procedure for preparation of compound G-4

A solution of G-3 (9.00 g, 23.8 mmol, 1.00 eq) in MeOH (100 mL) , degassed with Ar for 2 minutes. Then Pd/C (3.00 g, 23.8 mmol, 10.0 %purity, 1.00 eq) was added under Ar. The resulting mixture was degassed and purged with H ₂ for 3 times. The resulting mixture was stirred at 25 ℃ under H ₂ (15 psi) for 18 hrs. The mixture was filtered and the organic phase concentrated in vacuum to afford compound G-4 (8.00 g, crude) as a brown solid which was used for next step directly without purification.

LCMS: 348.2

General procedure for preparation of compound G-5

To a solution of G-4 (8.00 g, 23.0 mmol, 1.00 eq) in EtOAc (100 mL) was added DIPEA (11.9 g, 92.1 mmol, 16.0 mL, 4.00 eq) and T ₃P (29.3 g, 46.0 mmol, 27.3 mL, 50.0 %purity, 2.00 eq) under N ₂ at 0 ℃. The mixture was stirred at 20 ℃ for 18 hrs. To the mixture was added NaHCO ₃ solution (50 mL*2) , extracted with EtOAc (60 mL*2) , dried with MgSO _4, filtered and concentrated in vacuum. The residue was purified by flash silica gel chromatography (

80 g SepaFlash

Silica Flash Column, Eluent of 0～30%Ethyl acetate/Petroleum ether gradient @60 mL/min) . And compound G-5 (2.00 g, 6.07 mmol, 26.3%yield) was obtained as a brown oil.

LCMS: 330.1

General procedure for preparation of compound G-6

To a solution of G-5 (2.00 g, 6.07 mmol, 1.00 eq) in DMF (30 mL) was added K ₂CO ₃ (2.10 g, 15.1 mmol, 2.50 eq) . Then MeI (1.29 g, 9.11 mmol, 567 uL, 1.50 eq) was added dropwise at 0 ℃. The mixture was stirred at 25 ℃ for 18 hrs. To the mixture was added water (50 mL*2) , extracted with EtOAc (50 mL*2) , washed with brine (80 mL*2) , dried over Na ₂SO ₄, filter and concentrated in vacuum to afford compound G-6 (1.90 g, crude) as a brown oil which was used in next step without further purification.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 8.96 (dd, J=4.16, 1.59 Hz, 1 H) 8.42 (d, J=8.07 Hz, 1 H) 8.05 (d, J=9.05 Hz, 1 H) 7.64 (dt, J=8.74, 1.93 Hz, 2 H) 7.24 (dd, J=5.38, 2.57 Hz, 1 H) 4.42 (m, 3 H) 3.31 (s, 3 H) 1.33 (s, 9 H) .

LCMS: 344.1

General procedure for preparation of compound G-7

To a solution of G-6 (1.90 g, 5.53 mmol, 1.00 eq) in DCM (20 mL) was added m-CPBA (1.35 g, 6.64 mmol, 85.0 %purity, 1.20 eq) at 0-10℃. The mixture was stirred at 25 ℃ for 18 hrs. To the mixture was added Na ₂SO ₃, extracted with DCM (50 mL*2) water (50 mL*2) , dried with MgSO ₄, filter and concentrated in vacuum. The residue was purified by flash silica gel chromatography (

40 g

Silica Flash Column, Eluent of 0～40%THF/DCM@60 mL/min) . And compound G-7 (1.10 g, 3.06 mmol, 55.3%yield) was obtained as a yellow solid.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 8.63 (d, J=5.99 Hz, 1 H) 8.55 (d, J=9.41 Hz, 1 H) 7.91 (d, J=8.80 Hz, 1 H) 7.67 (d, J=9.41 Hz, 1 H) 7.56 (dd, J=8.80, 5.99 Hz, 1 H) 7.26 (br d, J=7.58 Hz, 1 H) 4.43 (m, 3 H) 3.30 (s, 3 H) 1.33 (s, 9 H) .

LCMS: 360.1

General procedure for preparation of compound G-8

A solution of G-7 (1.00 g, 2.78 mmol, 1.00 eq) in POCl ₃ (10.9 mL) was heated to 60 ℃. The mixture was stirred at 60 ℃ for 1 hr. The solution turned from yellow to brown. The mixture was concentrated to remove the solvent. To the residue was added sat. NaHCO ₃ (30 mL) , extracted with DCM (40 mL*2) , dried over MgSO ₄, filter and concentrated in vacuum. The mixture was used for next step without further purification.

To a solution of above mentioned intermediate (1.00 g, 3.60 mmol, 1.00 eq) in H2O (5 mL) and EtOAc (5 mL) was added NaHCO ₃ (302 mg, 3.60 mmol, 140 uL, 1.00 eq) , then Boc ₂O (943 mg, 4.32 mmol, 992 uL, 1.20 eq) was added at 25 ℃. The mixture was stirred at 25 ℃ for 1 hr. The mixture was extracted with EtOAc (5 mL*2) , the organic phase was dried with MgSO ₄, filtered and concentrated in vacuum. The residue was purified by flash silica gel chromatography (

40 g

Silica Flash Column, Eluent of 0～20% Ethyl acetate/Petroleum ether gradient @60 mL/min) . And compound G-8 (750 mg, 1.99 mmol, 55.1%yield) was obtained as a white solid.

¹H NMR (400 MHz, DMSO-d6) δ ppm 8.49 (d, J=8.88 Hz, 1 H) 7.98 (d, J=9.01 Hz, 1 H) 7.69 (dd, J=11.07, 8.94 Hz, 2 H) 7.23 (m, 1 H) 4.43 (m, 3 H) 3.34 (s, 3 H) 1.33 (s, 9 H) .

LCMS: 378.1

General procedure for preparation of compound G-9

To a solution of compound G-8 (80.0 mg, 212 umol, 1 eq) and morpholine (240 mg, 2.75 mmol, 242 uL, 13 eq) in i-PrOH (4.00 mL) was added TEA (64.3 mg, 635 umol, 88.4 uL, 3 eq) . The mixture was stirred at 100 ℃ for 65 h. And then the mixture was poured into water (5.00 mL) , extracted with EtOAc (5.00 mL*3) , concentrated in vacuum to give a yellow residue. The residue was purified by flash silica gel chromatography (

4 g

Silica Flash Column, Eluent of 14%Ethyl acetate/Petroleum ether@80mL/min) to give compound G-9 (81.6 mg, 190 umol, 89.9%yield) as a white solid.

LCMS: 429.2

General procedure for preparation of compound G-10

To a solution of compound G-9 (81.6 mg, 167 umol, 87.9%purity, 1 eq) in dioxane (1.00 mL) was added HCl/dioxane (4.00 M, 5.00 mL, 119 eq) at 0 ℃. The mixture was stirred at 25 ℃ for 2 h. And then the mixture was concentrated in vacuum to give compound G-10 (61.0 mg, crude, HCl) as a brown solid, which was directly used in the next step without further purification.

LCMS: 329.1

General procedure for preparation of compound 144

To a solution of HOBt (29.4 mg, 217 umol, 1.3 eq) , EDCI (41.7 mg, 217 umol, 1.3 eq) , DIPEA (64.8 mg, 501 umol, 87.4 uL, 3 eq) in DMF (3.00 mL) was added 5-benzyl-4H-1, 2, 4-triazole-3-carboxylic acid (44.2 mg, 217 umol, 1.3 eq) . The mixture was stirred at 25 ℃ for 10 min. Then compound G-10 (61.0 mg, 167 umol, 1 eq, HCl) was added. The mixture was stirred at 50 ℃ for 1 h and then was poured into water (5.00 mL) , extracted with EtOAc (5.00 mL*3) , concentrated in vacuum to give a yellow residue. The residue was purified by prep-HPLC (column: Phenomenex Gemini C18 250*50mm*10um; mobile phase: [water (0.05%ammonia hydroxide v/v) -ACN] ; B%: 30%-50%, 9 min) to give a compound 144 (28.7 mg, 55.9 umol, 33.4%yield, 100%purity) as a white solid.

¹H NMR: (400 MHz, DMSO-d ₆) δ ppm 14.11 -14.70 (m, 1 H) , 8.43 (br s, 1 H) , 8.08 (d, J=9.29 Hz, 1 H) , 7.59 (d, J=9.03 Hz, 1 H) , 7.45 (d, J=9.03 Hz, 1 H) , 7.20 -7.37 (m, 6 H) , 4.85 (dt, J=11.04, 8.03 Hz, 1 H) , 4.60 (br t, J=10.79 Hz, 1 H) , 4.45 (dd, J=9.79, 8.03 Hz, 1 H) , 4.12 (s, 2 H) , 3.60 -3.79 (m, 8 H) , 3.30 (s, 3 H) .

LC-MS: 514.1

Compounds in Table 6 below were prepared by following procedures that are the same as or similar to those for the preparation of compound 144 as described above.

Table 6:

Scheme XV below describes an illustrative synthetic scheme for preparation of compound 148 disclosed herein.

Scheme XV: Synthesis scheme for compound 148

General procedure for preparation of compound H-2

To a reaction mixture of KNO ₃ (3.09 g, 30.5 mmol, 1.00 eq) in H ₂SO ₄ (55.2 g, 563 mmol, 30 mL, 18.4 eq) was added compound H-1 (5.00 g, 30.6 mmol, 1.00 eq) in H ₂SO ₄ (55.2 g, 563 mmol, 30 mL, 18.4 eq) at 0 ℃ dropwise in 30 mins. The solution was stirred at 0 ℃ for 2 hrs. TLC (Eluent: PE: EA=3: 1) showed starting material was consumed completely. A new point (Rf = 0.42, desired product) were shown on TLC and desired compound was detected. The reaction mixture was poured into ice-water (300 mL, 1: 1) and stirred for 20 mins. The solution was adjusted pH to 7 -8 by Na ₂CO ₃ at 0 ℃. Solid was precipitated out. The suspension was filtered. The filter cake was checked by TLC (no desired product) and discarded. The aqueous phase was extracted with ethyl acetate (300 mL*3) . The combined organic phase was dried over anhydrous Na ₂SO ₄, filtered and concentrated in vacuum to afford compound H-2 (5.60 g, 26.85 mmol, 87.8%yield) as a yellow solid which was used for next step directly without purification.

¹H NMR: (400MHz, CHLOROFORM-d) δ ppm 9.36 (s, 1 H) , 8.74 (d, J=6.00 Hz, 1 H) , 8.11 (d, J=8.88 Hz, 1 H) , 7.71 (d, J=8.76 Hz, 1 H) , 7.56 (d, J=6.13 Hz, 1 H)

General procedure for preparation of compound H-3

To a solution of compound H-2 (5.00 g, 23.9 mmol, 1.00 eq) in DCM (100 mL) was added m-CPBA (7.00 g, 34.5 mmol, 85%purity, 1.44 eq) at 0 ℃ in potions. The solution was stirred at 20 ℃ for 16 hrs. And then the reaction mixture was poured into water (150 mL) and extracted with DCM (100 mL*3) . The combined organic phase was dried over anhydrous Na ₂SO ₄, filtered and concentrated in vacuum. The residue was purified by flash silica gel chromatography (

40 g

Silica Flash Column, Eluent of 0～50%Ethyl acetate/Petroleum ether gradient @40 mL/min) to afford desired product compound H-3 (5.00 g, 20.0 mmol, 83.6%yield, 90%purity) as a white solid.

¹H NMR: (400MHz, DMSO-d6) δ ppm 9.18 (d, J=1.76 Hz, 1 H) , 8.33 (dd, J=7.28, 1.76 Hz, 1 H) , 8.17 (d, J=9.04 Hz, 1 H) , 7.99 (d, J=9.03 Hz, 1 H) , 7.77 (d, J=7.28 Hz, 1 H) .

LC-MS: 224.9

General procedure for preparation of compound H-4

To a solution of compound H-3 (5.00 g, 20.0 mmol, 90%purity, 1.00 eq) in DCE (100 mL) was added POCl ₃ (15.4 g, 100 mmol, 9.31 mL, 5.00 eq) dropwise at 20 ℃. The solution was stirred at 70 ℃ for 3 hrs. The reaction mixture was added into ice-water (300 mL, 1: 1) dropwise in 10 mins at 0 ℃. The pH of the mixture was adjusted to 7-8 with NaHCO ₃ at 0 ℃. The suspension was extracted with DCM (300 mL*3) . The combined organic phase was dried over anhydrous Na ₂SO ₄, filtered and concentrated in vacuum. The residue was purified by flash silica gel chromatography (

40 g

Silica Flash Column, Eluent of 0 ～ 20%Ethylacetate/Petroleum ether gradient @40 mL/min) to afford compound H-4 (4.00 g, 14.8 mmol, 73.9%yield, 90%purity) as a yellow solid.

¹H NMR: (400MHz, DMSO-d6) δ ppm 8.59 (d, J=9.17 Hz, 1 H) , 8.54 (d, J=5.99 Hz, 1 H) , 8.14 (d, J=9.17 Hz, 1 H) , 7.74 (d, J=5.99 Hz, 1 H) .

LC-MS: 242.8

General procedure for preparation of compound H-5

To a solution of Boc-L-serine (2.28 g, 11.1 mmol, 1.50 eq) in DMF (40 mL) was added NaH (888 mg, 22.2 mmol, 60%purity, 3.00 eq) under N ₂ in portions in 30 mins. The reaction mixture was stirred at 0 ℃ for 1 hr. Compound H-4 (2.00 g, 7.41 mmol, 90%purity, 1.00 eq) was added into the reaction mixture in 30 mins at 0 ℃. The reaction mixture was stirred at 20 ℃ for 18 hrs before it was quenched by water (5 mL) at 0 ℃ and adjusted pH to 5-6 by HCl (2 N) . The solution was poured into water (100 mL) . The aqueous phase was extracted with EtOAc (100 mL*3) . The combined organic phase was washed with brine (100 mL*2) , dried over anhydrous Na ₂SO ₄, filtered and concentrated in vacuum. The reaction was scaled up as this condition for 2 times. The combined residue was purified by flash silica gel chromatography (

80 g

Silica Flash Column, Eluent of 0 ～ 20%MeOH/DCM@40 mL/min) to afford compound H-5 (6.00 g, 10.2 mmol, 68.8%yield, 70%purity) as a brown solid.

LC-MS: 412.0

General procedure for preparation of compound H-6

To a solution of compound H-5 (6.00 g, 10.2 mmol, 70%purity, 1.00 eq) in EtOH (140 mL) and H ₂O (40 mL) was added Fe (2.85 g, 51.0 mmol, 5.00 eq) and NH ₄Cl (1.09 g, 20.4 mmol, 2.00 eq) in portions at 20 ℃. The solution was stirred at 50 ℃ for 2 hrs before it was filtered. The filter cake was rinsed with EtOH (200 mL*3) . The rest of filter cake was checked by LCMS (no desired product) and discarded. The combined organic solution was concentrated under reduced pressure to afford desired product compound H-6 (6.00 g, 10.4 mmol, 66%purity, crude) as a brown solid which was used for next step directly without purification. LC-MS: 382.2

General procedure for preparation of compound H-7

To a solution of compound H-6 (6.00 g, 10.4 mmol, 66%purity, 1.00 eq) and TEA (3.15 g, 31.1 mmol, 4.33 mL, 3.00 eq) in THF (60 mL) was added T ₃P (9.90 g, 15.5 mmol, 9.25 mL, 50%purity in EtOAc, 1.50 eq) . The solution was stirred at 30 ℃ for 2 hrs. The reaction mixture was concentrated under reduced pressure at 40 ℃. The residue was added into water (200 mL) and extracted with DCM (200 mL*3) . The combined organic phase was washed with saturated solution of NaHCO ₃ in water (200 mL*2) , dried over anhydrous Na ₂SO ₄, filtered and concentrated in vacuum. The residue was purified by flash silica gel chromatography (

80 g

Silica Flash Column, Eluent of 0 ～30%Ethyl acetate/Petroleum ether gradient @60 mL/min) to afford compound H-7 (1.30 g, 3.57 mmol, 34.4%yield) as a yellow solid.

¹H NMR: (400MHz, CHLOROFORM-d) δ ppm 8.43 (br s, 1 H) , 8.37 (d, J=6.02 Hz, 1 H) , 8.27 (d, J=9.29 Hz, 1 H) , 7.72 (d, J=5.77 Hz, 1 H) , 7.50 (d, J=9.03 Hz, 1 H) , 5.56 (br d, J=6.27 Hz, 1 H) , 4.73 -4.89 (m, 2 H) , 4.42 -4.53 (m, 1 H) , 1.42 -1.46 (m, 10 H) .

LC-MS: 364.2

General procedure for preparation of H-8

To a mixture of compound H-7 (1.30 g, 3.57 mmol, 1.00 eq) and K ₂CO ₃ (741 mg, 5.36 mmol, 1.50 eq) in DMF (30 mL) was added MeI (608 mg, 4.29 mmol, 267 uL, 1.20 eq) dropwise at 20 ℃. The solution was stirred at 20 ℃ for 2 h before it was poured into water (50 mL) . The aqueous phase was extracted with ethyl acetate (50 mL*3) . The combined organic phase was washed with brine (50 mL*2) , dried over anhydrous Na ₂SO ₄, filtered and concentrated in vacuum to afford compound H-8 (1.00 g, crude) as a yellow solid which was used for next step directly without further purification.

LC-MS: 378.0

General procedure for preparation of compound H-9

To a solution of compound H-8 (0.40 g, 953 umol, 90%purity, 1.00 eq) and benzophenone imine (207 mg, 1.14 mmol, 192 uL, 1.20 eq) in toluene (6 mL) were added t-BuONa (183 mg, 1.91 mmol, 2.00 eq) , BINAP (119 mg, 190 umol, 0.20 eq) and Pd ₂ (dba) ₃ (87.2 mg, 95.3 umol, 0.10 eq) . The mixture was stirred at 100 ℃ for 16 hrs. The solvent was removed in reduced pressure at 50 ℃ and the crude product was purified by flash silica gel chromatography (

24g

Silica Flash Column, Eluent of 0～30%Ethyl acetate/Petroleum ether gradient @4 mL/min) to afford compound H-9 (130 mg, 249 umol, 26.1%yield) as a yellow solid.

LC-MS: 523.1

General procedure for preparation of compound H-10

A solution of compound H-9 (200 mg, 383 umol, 1.00 eq) in HCl/dioxane (4 M, 4 mL, 41.8 eq) was stirred at 20 ℃ for 2 hrs. The reaction was concentrated in reduced pressure at 50 ℃. The residue was washed with EtOAc (20 mL) to afford compound H-10 (130 mg, crude, HCl salt) as a yellow solid which was used for next step directly without further purification.

LC-MS: 302.9

General procedure for preparation of 148

To a solution of compound H-10 (130 mg, 442 umol, 1.00 eq, HCl salt) , 5-benzyl-4H-1, 2, 4-triazole-3-carboxylic acid (98.6 mg, 485 umol, 1.10 eq) and TEA (134 mg, 1.32 mmol, 184 uL, 3.00 eq) in DMF (5 mL) were added HOPO (58.8 mg, 529 umol, 1.20 eq) and EDCI (101 mg, 529 umol, 1.20 eq) . The solution was stirred at 60 ℃ for 2 hrs before it was concentrated in reduced pressure at 60 ℃. The residue was purified by flash silica gel chromatography (

4 g

Silica Flash Column, Eluent of 0～30%MeOH/DCM@35 mL/min) , and then by prep-HPLC (column: Phenomenex Gemini-NX 150*30mm*5um; mobile phase: [water (0.05%NH ₃H ₂O + 10mM NH ₄HCO ₃) -ACN] ; B%: 25%-45%, 8min) to afford compound 148 (35.1 mg, 78.0 umol, 17.7%yield, 98.6%purity) as a white solid.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 14.47 (br s, 1 H) , 8.47 (br s, 1 H) , 8.26 (d, J=9.29 Hz, 1 H) , 7.89 (d, J=6.02 Hz, 1 H) , 7.30 -7.38 (m, 3 H) , 7.22 -7.29 (m, 3 H) , 7.03 (s, 2 H) , 6.83 (d, J=6.02 Hz, 1 H) , 4.84 (dt, J=11.17, 8.09 Hz, 1 H) , 4.68 (br t, J=10.67 Hz, 1 H) , 4.46 (dd, J=9.79, 7.78 Hz, 1 H) , 4.12 (s, 2 H) , 3.26 (s, 3 H) .

LC-MS: 444.0

Compounds in Table 7 below were prepared by following procedures that are the same as or similar to those for the preparation of compound 148 as described above.

Table 7:

Scheme XVI below describes an illustrative synthetic scheme for preparation of compound 166 disclosed herein.

Scheme XVI: Synthesis scheme for compound 166

General procedure for preparation of compound I-2

To a solution of compound I-1 (51.3 g, 250 mmol, 1.10 eq) and in DMF (800 mL) was added in NaH (20.0 g, 500.00 mmol, 60%purity, 2.20 eq) at 0℃. The reaction was stirred at 0 ℃ for 2 hours. Boc-L-serine (50.00 g, 227 mmol, 27.9 mL, 1.00 eq) was added at 0℃. The reaction was stirred at 20 ℃ for 2 hours. And then the mixture was added into water (1000 mL) at 0 ℃ and stirred at 20℃ for 30 mins. The mixture was adjusted pH to ～5 by HCl (2 N) at 0-10℃. The aqueous phase was extracted with ethyl acetate (1000mL*3) . The combined organic phase was washed with brine (500 mL*2) , dried with anhydrous Na2SO4, filtered and concentrated in vacuum to afford compound I-2 (110 g, crude) as a yellow oil.

LCMS: 307.0

General procedure for preparation of compound I-3

To a solution of compound I-2 (100 g, 138 mmol, 56%purity, 1.00 eq) in EtOH (1000 mL) and H ₂O (500 mL) was added Fe (38.6 g, 691 mmol, 5.00 eq) and NH ₄Cl (36.9 g, 691 mmol, 5.00 eq) . The solution was stirred at 70 ℃ for 2 hours. The mixture was filtered and concentrated in reduced pressure at 50 ℃. The residue was dissolved in 1000 mL*3 THF. The mixture was filtered again, and the filtrate was concentrated in reduced pressure at 50 ℃ to afford compound I-3 (100 g, crude) as a brown oil which was used for next step directly without further purification.

LCMS: 318.9

General procedure for preparation of I-4

To a solution of compound I-3 (100 g, 125 mmol, 47%purity, 1.00 eq) , DIEA (48.5 g, 376 mmol, 65.4 mL, 3.00 eq) in EtOAc (1000 mL) was added T ₃P (159 g, 250 mmol, 149 mL, 50%purity, 2.00 eq) . The solution was stirred at 20 ℃for 2 hours before it was filtered and concentrated in reduced pressure. The residue was poured into water (1000 mL) . The aqueous phase was extracted with ethyl acetate (1000 mL*2) . The combined organic phase was washed with brine (1000 mL*2) , dried over anhydrous Na ₂SO ₄, filtered and concentrated in vacuum. The residue was purified by flash silica gel chromatography (

330 g

Silica Flash Column, Eluent of 0～20%Ethyl acetate/Petroleum ether gradient @80 mL/min) to afford compound I-4 (18.0 g, 50.4 mmol, 40.2%yield) as a yellow solid.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 9.66 (s, 1 H) , 7.52 (dd, J=7.78, 1.51 Hz, 1 H) , 7.10 -7.31 (m, 3 H) , 4.24 -4.47 (m, 3 H) , 1.36 (s, 9 H) .

LCMS: 301.1

General procedure for preparation of I-5

To a solution of I-4 (0.15 g, 420 umol, 1.00 eq) in dioxane (1 mL) was added HCl/dioxane (4 M, 2 mL, 19.0 eq) dropwise. The reaction was stirred at 20 ℃ for 2 hours. Solid was precipitated out. The reaction mixture was concentrated in reduced pressure at 50 ℃ to afford compound I-5 (120 mg, 49 umol, 97.3%yield, HCl salt) as a yellow solid which was used for next step directly without further purification.

LCMS: 259.0

General procedure for preparation of compound 166

To a solution of I-5 (120 mg, 409 umol, 1.00 eq, HCl salt) and 5-benzyl-4H-1, 2, 4-triazole-3-carboxylic acid (125 mg, 613 umol, 1.50 eq) in DMF (3 mL) was added DIEA (264 mg, 2.04 mmol, 356 uL, 5.00 eq) followed by T ₃P (390 mg, 613 umol, 364 uL, 50%purity, 1.50 eq) . The reaction was stirred at 20 ℃ for 2 hours and then quenched with water. The aqueous phase was extracted with ethyl acetate (10 mL*3) . The combined organic phase was washed with brine (10 mL*2) , dried with anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was purified by prep-HPLC (column: Phenomenex Gemini C18 250*50mm*10um; mobile phase: [water (0.05%ammonia hydroxide v/v) -ACN] ; B%: 30%-50%, 9min) to afford desired product compound 166 (58.9 mg, 133 umol, 32.6%yield) as a white solid.

¹H NMR: (400 MHz, DMSO-d6) δ ppm 9.50 -10.18 (m, 1 H) , 8.49 (br s, 1 H) , 7.55 (dd, J=8.03, 1.51 Hz, 1 H) , 7.16 -7.37 (m, 7 H) , 4.72 -4.83 (m, 1 H) , 4.65 (t, J=10.54 Hz, 1 H) , 4.46 (dd, J=9.79, 7.78 Hz, 1 H) , 4.13 (s, 2 H) .

LCMS: 443.9

Compounds in Table 8 below were prepared by following procedures that are the same as or similar to those for the preparation of compound 166 as described above.

Table 8

Example 2

In Vitro Cell Assays

The in vitro efficacy of the compounds disclosed herein as RIP1 inhibitors can be tested using cell lines in necroptosis assays.

Necroptosis Assay Using U937 Cells

The in vitro efficacy of RIP1 inhibitors were tested using human monocytic U937 cells (ATCC, CRL-1593.2) in the necroptosis assay. For the assay, RPMI-1640 medium was added to 96 well cell culture plate (50μL/well, including 10% FBS) . Then 0.5μL of the diluted compounds or 100%DMSO were transferred into plates containing RPMI-1640 medium. U937 cells at exponential phase were diluted to 5x105/mL in RPMI-1640 and seeded into assay plates (40μL/well) . Cells were mixed with the compounds thoroughly and then incubated at RT (room temperature) for 1 hour. hTNFα (human tumor necrosis factor, Novoprotein, c008) was added to the cell solution to achieve a final concentration of 20 ng/mL. QVD-Oph (Selleck chemical, S7311) , a pan-caspase inhibitor, was added to the cell solution to achieve a final concentration of 50μM . After 1-hour incubation at RT, the plates were incubated for 23 hours at 37℃, 5%CO2. On the next day, cells were lysed and ATP content (viability) was measured via the addition of 50 μL Cell Titer-Glo Luminescent Cell Viability reagent (Promega Corporation) . Plates were incubated for 10 minutes followed by luminescence detection on BioTek. The ability of tested compounds to rescue cells from TNFα/QVD induced necroptosis was represented as percent survival relative to 100%DMSO treated positive control wells. For concentration response experiments, normalized data were fitted and IC50 values determined using GraphPad Prism. All data are shown as mean ±standard deviation of the mean.

Necroptosis Assay Using L929 Cells

The in vitro efficacy of RIP1 inhibitors were tested using mouse L929 fibroblast cells (ATCC, CRL6364) in the necroptosis assay. For the assay, L929 cells at exponential phase were diluted to 1x10 ⁵/ml in MEM medium and seeded into assay plates (100μL/well) . Then 0.5μL of the diluted compounds or 100% DMSO were transferred into plates and incubated at RT for 1 hour. hTNFα (human tumor necrosis factor, Novoprotein, c008) was added to the cell solution to achieve a final concentration of 20 ng/mL. QVD-Oph (Selleck chemical, S7311) , a pan- caspase inhibitor, was added to the cell solution to achieve a final concentration of 50μM. After 1-hour incubation at RT, the plates were incubated for 23 hours at 37℃, 5%CO2. The next day, cells were lysed and ATP content (viability) was measured via the addition of 50 μL Cell Titer-Glo Luminescent Cell Viability reagent (Promega Corporation) . Plates were incubated for 10 minutes followed by luminescence detection on BioTek. The ability of tested compound to rescue cells from TNFα/QVD induced necroptosis was represented as percent survival relative to 100%DMSO treated positive control wells. For concentration response experiments, normalized data were fitted and IC50 values determined using GraphPad Prism. All data are shown as mean ± standard deviation of the mean.

TNF-induced Jurkat (FADD -/-) Cell Necrosis Assay

To a white 96-well cell culture plate was added RPMI-1640 medium (50 μL per well, including serum) . Then a 0.5μL solution containing the compound disclosed herein or control compound 7-Cl-O-Nec-1 and GSK2982772 (7-Cl-O-Nec-1 and GSK2982772 are known RIP1 kinase inhibitors) or a 0.5 μL 100% DMSO was added to each well, each compound has two replicate wells for each concentration.

Jurkat (FADD -/-) cells (human-derived peripheral blood leukemia T cell line, FADD gene knocked out) was cultured in vitro. After growing to logarithmic growth phase, the cells were collected and centrifuged at 1000 rpm for 5 minutes. The supernatant was discarded. The cells were resuspended in the fresh RPMI-1640 medium and the cell concentration was adjusted to 5×10 ⁵/mL. The cell suspension was added to the culture plate containing the compounds to be tested (40 μL per well) .

In the stimulation group, 10μL of TNF (tumor necrosis factor, final concentration 10ng/mL) diluted with cell culture medium was added to each well, placed in a cell culture incubator (37℃, 5%CO 2) for 20 or 24 hours, and 50μL of Cell Titer-Glo solution was added to each well. After incubation at room temperature for 10 minutes, BioTek plate reader was used to detect the luminescence value to measure the intracellular ATP level. The ability of the tested compound in rescuing TNF-induced cell necrosis was represented as percent survival relative to 100%DMSO treated positive control wells. Graphpad Prism statistical software was used to calculate the compound’s IC50.

The in vitro assay data obtained from the necroptosis assays described above are shown in Table 9 below.

Table 9

Claims

A compound of formula I or a pharmaceutically acceptable salt thereof:

wherein

R ₁ is H, F, CN, CF ₃, or Cl,

R ₂ is H, -CH ₃, -CD ₃, or cyclopropyl,

A ring is a bicyclic fused ring system represented by
in which N ring is fused to the amide-containing seven-membered ring,

N ring is a pyridine, pyrazine, or phenyl optionally substituted with one or two groups selected from F and Cl,

M is a 5 or 6 membered heteroaryl or phenyl each of which is optionally substituted by one or two groups selected from oxo, Cl, -O-C _1-3 alkyl, OH, CN, C _1-3 alkyl, -S-C _1-3 alkyl, phenyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and-NR _aR _b,

W is independently selected from -C _1-3 alkyl, oxo, -OH, -COOC _1-3 alkyl, C _3-5 cycloalkyl, haloC _1-3 alkyl, -O-C _1-3 alkyl, Cl, and CN,

R _a is H, CH ₃, or CH ₂CH ₃,

R _b is selected from -C _1-3 alkyl, haloC _1-3 alkyl, -CH ₂-C _3-5 cycloclkyl, -CH ₂-phenyl, optionally substituted 5 or 6 membered heterocyclyl, C _3-5 cycloalkyl, - (CH ₂) _2-3OH, - (CH ₂) _2-3OCH ₃, wherein the optional substituent is selected from C _1-3 alkyl and C _3-5 cycloalkyl,

X is O or CH ₂,

B is
in which the amide nitrogen is attached to the amide-containing seven membered ring, X ₁ is CH or N, R _c is selected from H, -CH ₃, -CD ₃, -CH ₂CF ₃, -CH=CH ₂, and cyclopropyl, and R _c is attached to one of the nitrogen of the 5-membered ring,

L is C=O, CHCH ₃, C (CH ₃) ₂, O, CHOH, CH ₂ or

provided that when B is
N ring is a non-substituted phenyl, and (1) R _c is not -CH=CH ₂ or cyclopropyl, (2) R ₂ is not cyclopropyl, and (3) L is not
then M ring is substituted by one or two groups selected from oxo, S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b excluding NH ₂.
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein M ring is an oxazoline, pyrimidine, pyridine, or pyrazine optionally substituted by one or two groups selected from oxo, C _1-3 alkyl, -S-C _1-3 alkyl, 5 to 12 membered heterocycyl optionally substituted with one or more W, and -NR _aR _b.
The compound or a pharmaceutically acceptable salt thereof of claim 1 or 2, wherein the 5 to 12 membered heterocycyl optionally substituted with one or two W is selected from pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, 5, 6, 7, 8-tetrahydroimidazo [1, 2-a] pyrazinyl, tetrahydro-2H-pyranyl, 8-oxa-3-azabicyclo [3.2.1] octanyl, 3-oxa-9-azaspiro [5.5] undecanyl, 7-oxa-2-azaspiro [3.5] nonanyl, and 2-oxa-7-azaspiro [3.5] nonanyl, each of which is optionally substituted with one or two W.
The compound or a pharmaceutically acceptable salt thereof of any of claims 1-3, when the 5 to 12 membered heterocycyl optionally substituted with one or two W is a nitrogen-containing 5 to 12 membered heterocycyl, the 5 to 12 membered heterocycyl is attached to M ring through the nitrogen.
The compound or a pharmaceutically acceptable salt thereof of any of claims 1-4, wherein the optionally substituted 5 or 6 membered heterocyclyl for R _b is optionally substituted pyrrolidinyl, piperazinyl, morpholinyl or piperidinyl.
The compound or a pharmaceutically acceptable salt thereof of any of claims 1-5, wherein B is
The compound or a pharmaceutically acceptable salt thereof of any of claims 1-6, wherein L is CH ₂ or
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein A ring is
wherein R ₃ is selected from H, -S-C _1-3 alkyl, phenyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, R ₄ is H, F, or Cl.
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein A ring is
wherein R ₅ is selected from H, OH, Cl, -O-C _1-3 alkyl, -S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, R ₄ is H, F, or Cl.
The compound or a pharmaceutically acceptable salt thereof of claim 9, wherein R ₅ is selected from C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b.
The compound or a pharmaceutically acceptable salt thereof of claim 10, wherein R ₅ is selected from 5 or 6 membered heterocyclyl and -NH-C _3-5 cycloalkyl.
The compound or a pharmaceutically acceptable salt thereof of claim 11, wherein R ₅ is selected from -NH-cyclopropyl and morpholinyl.
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein A ring is
R ₄ is H, F, or Cl, R ₆ is selected from H, CN, -S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b, R ₄ is H, F, or Cl.
The compound or a pharmaceutically acceptable salt thereof of claim 13, wherein R ₆ is selected from H, CN, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b.
The compound or a pharmaceutically acceptable salt thereof of claim 14, wherein R ₆ is selected from H, CN, 5 or 6 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b.
The compound or a pharmaceutically acceptable salt thereof of claim 15, wherein, R ₆ is selected from H, CN, morpholinyl, piperidinyl, pyrrolidinyl, -NH-cyclopropyl, -NH (CH ₂) _2-3OH, -NH (CH ₂) _2-3OCH ₃, piperazinyl, -NHCH ₂-cyclopropyl, wherein the piperidinyl, pyrrolidinyl, and piperazinyl, are optionally substituted with -C _3-5 cycloalkyl or -C _1-3 alkyl.
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein A ring is
Z is CH or N, R ₇ and R ₈ are independently H, F, or Cl, provided that at least one of R ₇ and R ₈ is not H, and R ₉ is selected from H, -S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b.
The compound or a pharmaceutically acceptable salt thereof of claim 17, wherein R ₉ is selected from C _3-6 cycloalkyl optionally substituted with one or two W, 5 or 6 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b.
The compound or a pharmaceutically acceptable salt thereof of claim 18, wherein R ₉ is selected from morpholinyl, piperidinyl, pyrrolidinyl, -NH-cyclopropyl, -NH-tetrahydro-2H-pyranyl, -NH (CH ₂) _2-3OH, -NH (CH ₂) _2-3OCH ₃, piperazinyl, -NHCH ₂-cyclopropyl, wherein the piperidinyl, pyrrolidinyl, and piperazinyl, are optionally substituted with -C _3-5 cycloalkyl or -C _1-3 alkyl.
The compound or a pharmaceutically acceptable salt thereof of claim 19, wherein R ₉ is H or -C _1-3 alkyl.
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein A ring is
wherein R ₁₀ is selected from H, -C _1-3 alkyl, -S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b.
The compound or a pharmaceutically acceptable salt thereof of claim 21, wherein R ₁₀ is H or C _3-5 cycloalkyl.
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein A ring is
R ₁₁ is selected from H, -C _1-3 alkyl, -S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b.
The compound or a pharmaceutically acceptable salt thereof of claim 23, wherein R ₁₁ is selected from 5 to 6 membered heterocycyl optionally substituted with one or two W and -NR _aR _b.
The compound or a pharmaceutically acceptable salt thereof of claim 24, wherein R ₁₁ is morpholinyl, piperazinyl, -NH-cyclopropyl, or -NH-tetrahydro-2H-pyranyl, wherein piperazinyl is optionally substituted with C _3-5 cycloalkyl.
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein A ring is
R ₁₂ is selected from H, CN, -C _1-3 alkyl, -S-C _1-3 alkyl, C _3-6 cycloalkyl optionally substituted with one or two W, 5 to 12 membered heterocycyl optionally substituted with one or two W, and -NR _aR _b.
The compound or a pharmaceutically acceptable salt thereof of claim 26, wherein R ₁₂ is selected from -NR _aR _b and 5 to 6 membered heterocycyl optionally substituted with one or two W.
The compound or a pharmaceutically acceptable salt thereof of claim 27, wherein R ₁₂ is -NH (CH ₂) _2-3OH, -NH (CH ₂) _2-3-OCH ₃, morpholinyl, -NHCH ₂-cyclopropyl, piperazinyl, -NH-cyclopropyl, or -NH-tetrahydro-2H-pyranyl, wherein piperazinyl is optionally substituted with C _1-3 alkyl or C _3-5 cycloalkyl.
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein A ring is
Z is CH or N.
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein A ring is
R ₁₃ is C _1-3 alkyl or C _3-5 cycloalkyl.
The compound or a pharmaceutically acceptable salt thereof of claim 1, wherein A ring is selected from
The compound or a pharmaceutically acceptable salt thereof of any of claims 8-31, wherein B is
The compound or a pharmaceutically acceptable salt thereof of any of claims 8-32, wherein L is CH ₂ or
A compound selected from

or a pharmaceutically acceptable salt thereof.
A compound selected from

or a pharmaceutically acceptable salt thereof.
A compound selected from

or a pharmaceutically acceptable salt thereof.
A pharmaceutical composition comprising a compound of any of claims 1-36 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.
A method of inhibiting RIP1-kinase in a subject in need thereof, comprising administering a therapeutically effective amount of a composition of claim 37.
A method of treating a disease or disorder mediated by RIP1-kinase in a subject in need thereof, comprising administering a therapeutically effective amount of a composition of claim 37.
The method of claim 39, wherein the disease or disorder mediated by RIP1-kinase is selected from cancer and inflammatory diseases.
The method of claim 39, wherein the disease or disorder is selected from inflammatory bowel diseases, Crohn's disease, ulcerative colitis, psoriasis, retinal detachment, retinitis pigmentosa, macular degeneration, pancreatitis, atopic dermatitis, arthritis (including rheumatoid arthritis, osteoarthritis, spondylarthritis, gout, systemic onset juvenile idiopathic arthritis (SoJIA) , psoriatic arthritis) , systemic lupus erythematosus (SLE) , Sjogren's syndrome, systemic scleroderma, anti-phospholipid syndrome (APS) , vasculitis, liver damage/diseases (non-alcohol steatohepatitis, alcohol steatohepatitis, autoimmune hepatitis autoimmune hepatobiliary diseases, primary sclerosing cholangitis (PSC) , acetaminophen toxicity, hepatotoxicity) , kidney damage/injury (nephritis, renal transplant, surgery, administration of nephrotoxic drugs e.g. cisplatin, acute kidney injury (AKI) ) , Celiac disease, autoimmune idiopathic thrombocytopenic purpura, transplant rejection, ischemia reperfusion injury of solid organs, sepsis, systemic inflammatory response syndrome (SIRS) , cerebrovascular accident (CVA, stroke) , myocardial infarction (MI) , atherosclerosis, Huntington's disease, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS) , spinal muscular atropy (SMA) , allergic diseases (including asthma and atopic dermatitis) , multiple sclerosis, type I diabetes, Wegener's granulomatosis, pulmonary sarcoidosis, Behcet's disease, interleukin-1 converting enzyme (ICE, also known as caspase-1) associated fever syndrome, chronic obstructive pulmonary disease (COPD) , tumor necrosis factor receptor-associated periodic syndrome (TRAPS) , periodontitis, linear ubiquitin chain assembly complex (LUBAC) deficiency syndrome, hematological and solid organ malignancies, bacterial infections and viral infections (such as tuberculosis and influenza) , and Lysosomal storage diseases.
The method of claim 39, wherein the disease or disorder is selected from inflammatory bowel disease, Crohn's disease, ulcerative colitis, multiple sclerosis, arthritis, rheumatoid arthritis, osteoarthritis, and psoriasis.
The method of any of claims 39-42, further comprising administering to the subject in need thereof an additional known anti-inflammatory agent.
The method of claim 43, wherein the additional known anti-inflammatory agent is selected from corticosteroids, 5-aminosalicyclic acid, tofacitinib, rituximab, and adalimumab.