RU2800652C2 - Bicyclic compound as rip-1 kinase inhibitor and its use - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to a compound having formula (I), or a stereoisomer or a pharmaceutically acceptable salt thereof, which has RIP-1 kinase inhibitory activity. In formula (I), T₁ is selected from N and CR_1t; T₂ is selected from N and CR_2t; T₃ is CR_3t; T₄ is CR_4t; E₁ is selected from C(R_1e)₂ and O; cycle A is selected from 1,2,4-triazolyl, where 1,2,4-triazolyl, optionally substituted with 1, 2 or 3 halogens or C_1-3 alkyls; cycle B is selected from phenyl and L is selected from a single bond, O and C_1-3 alkylene; R₁ is selected from H and C_1-3 alkyl, where C_1-3 alkyl is optionally substituted with 1, 2 or 3 R_b substituents; R_1t, R_2t, R_3t and R_4t are each independently selected from H, F, Cl, Br, I, CN, C_1-3 alkyl, COOH and -C(=O)NH₂; R_1e is each independently selected from H and C_1-3 alkyl; R₂ is each independently selected from H, F, Cl, Br, I, CN, C_1-3 alkyl and C_1-3 alkoxy, where C_1-3 alkyl and C_1-3 alkoxy are optionally substituted with 1, 2 or 3 substituents R_f; n is 1, 2, 3, 4 or 5; R_b and R_f are each independently selected from F, Cl, Br, I and D. The invention also relates to particular compounds and to the use of the said compounds in the manufacture of a medicinal product for the treatment of a disease associated with RIP-1 kinase.

EFFECT: obtaining bicyclic compound as RIP-1 kinase inhibitor.

16 cl, 8 tbl, 42 ex

Description

The field of technology to which the invention belongs

The present invention relates to a compound having formula (I), or an isomer or pharmaceutically acceptable salt thereof, and their use in the manufacture of a medicament for the treatment of diseases associated with RIP-1 (receptor interacting protein) kinase.

Prior Art

RIP1 is an important kinase that controls cell necrosis and apoptosis. RIP1 is also involved in various inflammatory response signaling pathways and can be stimulated by ligands from the TNF and TLR families to trigger an inflammatory response (Weinlich, R. Nat. Rev. Mol. Cell. Biol. 2017, 18, 127; Pasparakis M. et al., Nature 2015, 517, 311; Berger, S. B. Cell Death Discovery, 2016, 2, e16056). RIP1 triggers a number of inflammatory response signaling pathways [Fas ligand, TNF family apoptosis-inducing ligand (TRAIL), TLR3 and TLR4] via TNF receptor 1 (Lukens J. R., et al. Nature, 2013, 498, 224). Inhibition of RIP1 activity can be used to treat many inflammatory diseases.

Brief description of the invention

The present invention describes a compound having formula (I), or an isomer or pharmaceutically acceptable salt thereof,

Where:

T ₁ is selected from N and CR _1t ;

T ₂ is selected from N and CR _2t ;

T ₃ is selected from N and CR _3t ;

T ₄ is selected from N and CR _4t ;

E ₁ is selected from C(R _1e ) ₂ , O, C(=O), S and NR _2e ;

Ring A is selected from 1,2,4-triazolyl, 1,2,3-triazolyl, imidazolyl, isoxazolyl, 1,3,4-oxadiazolyl, , tetrazolyl, pyridyl and , where 1,2,4-triazolyl, 1,2,3-triazolyl, imidazolyl, isoxazolyl, 1,3,4-oxadiazolyl, , tetrazolyl, pyridyl and optionally substituted with 1, 2 or 3 halo or C _1-3 alkyl;

Cycle B is selected from phenyl and ;

L is selected from a single bond, O, C(=O), S, NH, and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2, or 3 _{R a} substituents;

R ₁ is selected from H and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _b substituents;

R _1t , R _2t , R _3t and R _4t are each independently selected from H, F, Cl, Br, I, OH, CN, NH ₂ , C _1-3 alkyl, COOH and -C(=O)NH ₂ where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _c substituents;

R _{1e is} each independently selected from H, F, Cl, Br, I, OH, CN, NH ₂ and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _d substituents;

R _2e is selected from H and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _e substituents;

R ₂ is each independently selected from H, F, Cl, Br, I, OH, NH ₂ , CN, C _1-3 alkyl and C _1-3 alkoxy, where C _1-3 alkyl and C _1-3 alkoxy the group is optionally substituted with 1, 2 or 3 R _f substituents;

n is 1, 2, 3, 4 or 5;

R _a , R _b , R _c , R _d , R _e and R _f are each independently selected from F, Cl, Br, I, OH, CN, NH ₂ and D.

The present invention describes a compound having formula (I), or an isomer or pharmaceutically acceptable salt thereof,

,

Where

T ₁ is selected from N and CR _1t ;

T ₂ is selected from N and CR _2t ;

T ₃ is selected from N and CR _3t ;

T ₄ is selected from N and CR _4t ;

E ₁ is selected from C(R _1e ) ₂ , O, C(=O), S and NR _2e ;

Ring A is selected from 1,2,4-triazolyl, 1,2,3-triazolyl, imidazolyl, isoxazolyl, 1,3,4-oxadiazolyl and , where 1,2,4-triazolyl, 1,2,3-triazolyl, imidazolyl, isoxazolyl, 1,3,4-oxadiazolyl and optionally substituted with 1, 2 or 3 halo or C _1-3 alkyl;

Cycle B is phenyl;

L is selected from a single bond, O, C(=O), S, NH, and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2, or 3 _{R a} substituents;

R ₁ is selected from H and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _b substituents;

R _1t , R _2t , R _3t and R _4t are each independently selected from H, F, Cl, Br, I, OH, CN, NH ₂ and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 substituents R _c ;

R _{1e is} each independently selected from H, F, Cl, Br, I, OH, CN, NH ₂ and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _d substituents;

R _2e is selected from H and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _e substituents;

R ₂ is each independently selected from H, F, Cl, Br, I, OH, NH ₂ , CN and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _f substituents;

n is 1, 2, 3, 4 or 5;

R _a , R _b , R _c , R _d , R _e and R _f are each independently selected from F, Cl, Br, I, OH, CN, NH ₂ and D.

The present invention describes a compound having formula (I), or an isomer or pharmaceutically acceptable salt thereof,

,

Where

T ₁ is N or CR _1t ;

T ₂ is N or CR _2t ;

T ₃ is N or CR _3t ;

T ₄ is N or CR _4t ;

E ₁ is C(R _1e ) ₂ , O, C(=O), S, or NR _2e ;

Cycle A is a 5-membered heteroaryl optionally substituted with C _1-3 alkyl;

Cycle B is phenyl or 6-membered heteroaryl;

L is O, C(=O), S, NH or C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _a substituents;

R ₁ is selected from H and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _b substituents;

R _1t , R _2t , R _3t and R _4t are each independently selected from H, F, Cl, Br, I, OH, CN, NH ₂ and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 substituents R _c ;

R _{1e is} each independently selected from H, F, Cl, Br, I, OH, CN, NH ₂ and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _d substituents;

R _2e is selected from H and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _e substituents;

R ₂ is each independently selected from H, F, Cl, Br, I, OH, NH ₂ , CN and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _f substituents;

n is 1, 2, 3, 4 or 5;

R _a , R _b , R _c , R _d , R _e and R _f are each independently selected from F, Cl, Br, I, OH, CN and NH ₂ .

In some embodiments described herein, R ₁ above is selected from H and CH ₃ , where CH ₃ is optionally substituted with 1, 2, or 3 R _b substituents, and other variables are as defined herein.

In some embodiments described herein, R ₁ as defined above is selected from H, CH ₃ and CD ₃ and other variables are as defined herein.

In some embodiments described herein, R ₁ as defined above is selected from H and CH ₃ and other variables are as defined herein.

In some embodiments described herein, the above R _1t , R _2t , R _3t and R _4t are each independently selected from H, F, Cl, Br, I, OH, CN, NH ₂ , CH ₃ , COOH and - C(=O)NH ₂ , where CH ₃ is optionally substituted with 1, 2, or 3 R _c substituents, and other variables are as defined herein.

In some embodiments described herein, the above R _1t , R _2t , R _3t and R _4t are each independently selected from H, F, Cl, Br, I, OH, CN, NH ₂ and CH ₃ where CH ₃ optionally substituted with 1, 2, or 3 R _c substituents, and other variables are as defined herein.

In some embodiments described herein, the above R _1t , R _2t , R _3t and R _4t are each independently selected from H, F, Cl, Br, I, OH, CN, NH ₂ and CH ₃ , and other variables have the meanings given in this text.

In some embodiments described herein, the above R _1e is each independently selected from H, F, Cl, Br, I, OH, CN, NH _{2 ,} and CH ₃ , and other variables are as defined herein.

In some embodiments described herein, R _2e as defined above is selected from H and CH ₃ and other variables are as defined herein.

In some embodiments described in this text, the above E ₁ represents CH ₂ , O, C(=O), S or NH, and other variables have the meanings specified in this text.

In some embodiments described herein, the above R ₂ is each independently selected from H, F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ and OCH ₃ where CH ₃ and OCH ₃ are optionally substituted with 1 , 2 or 3 substituents R _f , and other variables have the meanings specified in the present text.

In some embodiments described herein, the above R ₂ is each independently selected from H, F, Cl, Br, I, OH, NH ₂ , CN, CH ₃ , OCH ₃ , CF ₃ and OCF ₃ , and other variables have the meanings given in this text.

In some embodiments described herein, the above R ₂ is each independently selected from H, F, Cl, Br, I, OH, NH ₂ and CN, and other variables are as defined herein.

In some embodiments described herein, Cycle A above is selected from , , , , , , , , , , And , Where , , , , , , , , , And optionally substituted with 1, 2 or 3 halogens or C _1-3 alkyls, and other variables have the meanings given in this text.

In some embodiments described herein, Cycle A above is selected from , , , , , , And .

In some embodiments described herein, Cycle A above is pyrrolyl, imidazolyl, pyrazolyl, furyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, or 1, 3,4-thiadiazolyl, where pyrrolyl, imidazolyl, pyrazolyl, furyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl or 1,3,4-thiadiazolyl is optionally substituted with C _{1 -3} alkyl, and other variables have the meanings given in this text.

In some embodiments described herein, L as defined above is selected from a single bond, -CH ₂ - and -O-, and other variables are as defined herein.

In some embodiments described herein, L as defined above is selected from a single bond and -CH ₂ -, and other variables are as defined herein.

In some embodiments described herein, Cycle A above is , and other variables have the meanings given in this text.

In some embodiments described herein, Cycle B as defined above is phenyl or pyridyl and other variables are as defined herein.

The present invention includes embodiments implemented by combining any of the above variables.

In some embodiments described herein, the above compound is selected from

And

Where

n, T ₁ , T ₂ , T ₃ , R ₁ , R ₂ and R are as defined herein;

the carbon atom with “*” is a chiral carbon atom that exists in the form of an individual (R) or (S) enantiomer or a mixture enriched in one enantiomer;

or an isomer or pharmaceutically acceptable salt thereof.

In some embodiments described herein, the above compound is selected from

And

Where

n, T ₁ , T ₂ , T ₃ , R ₁ , R ₂ and R are as defined herein;

or an isomer or pharmaceutically acceptable salt thereof.

The present invention also describes compounds having the following formulas, or their isomers or pharmaceutically acceptable salts:

In some embodiments described herein, the above compound is selected from

And

or their isomers or pharmaceutically acceptable salts.

The present invention also describes the use of the above compound, or an isomer or pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of diseases associated with RIP-1 kinase.

The technical effect of the invention

As a novel RIP-1 kinase inhibitor, the compound of the present invention has an active inhibitory effect on programmed necrosis (necroptosis) of U937 cells induced by TNFα/QVD-OPh, and has a good inhibitory effect, showing a decrease in body temperature in mice in a TNF- triggered systemic inflammatory response syndrome.

Definitions and terms

Unless otherwise indicated, the following terms and phrases used in this text have the following meanings. Private terms or phrases should not be considered vague or obscure in the absence of a specifying definition, but should be understood in their usual meaning. When a trade name is used in this text, it means the corresponding material or active ingredient.

The term "pharmaceutically acceptable" is used in this text to refer to those compounds, materials, compositions and/or dosage forms that are suitable for use in contact with human or animal tissues within the framework of current medical knowledge, do not have excessive toxicity, irritant action, do not cause allergic reactions or other problems or complications, and have a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salt" means a salt of a compound of the present invention, which is obtained by reacting a compound containing a certain substituent disclosed herein with a relatively non-toxic acid or base. When the compounds of the present invention contain a relatively acidic functional group, a salt with a base can be prepared by contacting the neutral form of the compound with a sufficient amount of base, without solvent or in a suitable neutral solvent. The pharmaceutically acceptable base salt includes a sodium, potassium, calcium, ammonium, organic amine or magnesium salt, or similar salts. When the compounds of the present invention contain a relatively basic functional group, an acid salt can be prepared by contacting the neutral form of the compound with a sufficient amount of acid, without solvent or in a suitable neutral solvent. A pharmaceutically acceptable acid salt includes a salt with an inorganic acid, where the inorganic acid includes, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphoric acid, and etc.; and a salt with an organic acid, where the organic acid includes, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid , p-toluenesulfonic acid, citric acid, tartaric acid and methanesulfonic acid and the like; and an amino acid salt (such as arginine and the like), and an organic acid salt such as glucuronic acid and the like. Some compounds of the present invention contain both a basic and an acidic functional group and can be converted into both an acid salt and a base salt.

A pharmaceutically acceptable salt of the present invention can be obtained from a parent compound containing an acidic or basic moiety using standard chemical methods. In general, such a salt can be prepared by reacting the free acidic or basic form of the compound with a stoichiometric amount of a suitable base or acid, in water or in an organic solvent, or mixtures thereof.

Unless otherwise indicated, the term "isomer" includes geometric isomers, cis or trans isomers, stereoisomers, enantiomers, optical isomers, diastereomers and tautomers.

The compounds of the present invention may be present in a particular geometric or stereoisomeric form. The present invention covers all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereoisomers, (D)-isomers, (L)-isomers and racemic mixtures and other mixtures, for example an enantiomerically or diastereomerically enriched mixture, all of which are within the scope of the present invention. A substituent such as alkyl may contain an additional asymmetric carbon atom. All such isomers and mixtures thereof are within the scope of the present invention.

Unless otherwise indicated, the term "enantiomer" or "optical isomer" means stereoisomers that are mirror images of each other.

Unless otherwise indicated, the term "cis-trans isomer" or "geometric isomer" reflects the absence of the possibility of free rotation around a double bond or a single bond between the carbon atoms that are part of the cycle.

Unless otherwise indicated, the term "diastereomer" means stereoisomers that contain two or more chiral centers in the molecule and are not mirror images of each other.

Unless otherwise indicated, "(D)" or "(+)" means the dextroisomer, "(L)" or "(-)" means the levorotatory isomer, and "(DL)" or "(±)" means the racemate.

Unless otherwise noted, a solid wedge bond ( ) and dotted wedge connection ( ) denote the absolute configuration of the stereocenter; continuous feedforward ( ) and a dotted direct link ( ) denote the relative configuration of the stereocenter; wavy line ( ) means a continuous wedge-shaped connection ( ) or dotted wedge connection ( ); or wavy line ( ) means a continuous direct connection ( ) and a dotted direct link ( ).

Unless otherwise noted, when a compound has a double bond, such as a carbon-carbon double bond, a carbon-nitrogen double bond, and a nitrogen-nitrogen double bond, and two different substituents are attached to each atom of the double bond (in a double bond involving an atom nitrogen, the free pair of lone electrons at the nitrogen atom is considered one of the substituents), the compound is the (Z) isomer, (E) isomer, or a mixture of these two isomers of the compound, if the atoms at the double bond in the compound are connected to the substituents by a wavy line ( ). For example, a compound having the formula (A) below means that the compound is a single isomer of formula (A-1) or formula (A-2) or a mixture of two isomers of formula (A-1) and formula (A-2 ); and a compound having formula (B) below means that the compound is a single isomer of formula (B-1) or formula (B-2) or a mixture of two isomers of formula (B-1) and formula (B-2) . A compound having the formula (C) shown below means that the compound is a single isomer of formula (C-1) or formula (C-2), or a mixture of two isomers of formula (C-1) and formula (C-2) .

(A) (A-1) (A-2)

(B) (B-1) (B-2)

(C) (C-1) (C-2)

The compounds of the present invention may be present in a particular form. Unless otherwise indicated, the terms "tautomer" or "tautomeric form" mean that different functional groups are in dynamic equilibrium at room temperature and can rapidly transform into each other. If tautomers are possible (as in solution, for example), a tautomeric equilibrium can be established. For example, protic tautomers (also known as prototropic tautomers) include proton transfer interconversions such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers involve interconversions by rearranging some of the bonding electrons. A particular example of keto-enol tautomerization is the interconversion between two tautomers - pentan-2,4-dione and 4-hydroxypent-3-en-2-one.

Unless otherwise indicated, the term "enriched in one isomer", "isomerically enriched", "enriched in one enantiomer" or "enantiomerically enriched" means that the content of one isomer or enantiomer is less than 100% and the content of one isomer or enantiomer is 60% or more, or 70% or more, or 80% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, 98% or more, 99% or more, 99.5% or more, 99.6% or more, 99.7% or more, 99.8% or more, or 99.9% or more.

Unless otherwise indicated, the term "isomeric excess" or "enantiomeric excess" means the difference between the relative percentages of two isomers or two enantiomers. For example, if one isomer or enantiomer is present at 90% and the second isomer or enantiomer is present at 10%, the isomeric or enantiomeric excess (ee value) is 80%.

The optically active (R)- and (S)-isomer, or D and L isomer, can be obtained using chiral synthesis techniques or chiral reagents or other well-known methods. If one kind of enantiomer of a particular compound of the present invention is desired, the pure target enantiomer can be obtained by asymmetric synthesis or by using a chiral derivatizing agent, followed by separation of the resulting diastereomeric mixture and cleavage of the derivatizing group. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), the compound is reacted with a suitable optically active acid or base to form a diastereomeric salt, which is then separated into the individual diastereomers by methods known in the art, yielding a pure enantiomer. In addition, the enantiomer and diastereomer can be isolated using chiral stationary phase chromatography, optionally using chemical derivatization techniques (eg, obtaining a carbamate from an amine).

The compounds of the present invention may have an unnatural isotopic ratio of atoms at one or more of the atoms constituting the compound. For example, a compound may be labeled with a radioactive isotope such as tritium ( ³ H), iodine-125 ( ¹²⁵ I), or C-14 ( ¹⁴ C). In another example, hydrogen can be replaced with a heavier isotope of hydrogen, forming a deuterated drug. The bond between deuterium and carbon is stronger than between ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs have the advantage of reduced toxic side effects, improved drug stability, improved potency, and increased drug half-life. All changes in the isotopic composition of the compounds of the present invention, regardless of the radioactivity of the isotopes, are included within the scope of the present invention.

The term "optional" or "optional" means that the event or condition described below may occur, but is not required, and that the term includes cases where the specified event or condition is met and cases where the specified event or condition is not met.

The term "substituted" means that one or more hydrogen atoms on a particular atom are replaced by a substituent, including deuterium and hydrogen variants, subject to the valency of that atom and the stability of the resulting substituted compound. When a substituent is an oxo group (i.e. =O), this means that two hydrogen atoms have been replaced. Positions on an aromatic ring cannot be replaced by an oxo group. The term "optionally substituted" means that the atom may or may not have substituent(s), unless otherwise indicated, the type and number of substituents may be different, subject to their chemical possibility.

When a variable (such as R) occurs more than once in a construct or in a join structure, the value of that variable in each instance is independent of the others. Thus, for example, if a group has 0 to 2 R substituents, the group may optionally have up to two R substituents, where the R value is independent in each case. In addition, a combination of substituents and/or variants thereof is only permitted if the combination results in a stable compound.

When the number of linking groups is 0, for example in -(CRR) ₀ -, this means that this linking group is a single bond.

When one of the variables is a simple link, it means that the two groups connected by this simple link are directly related. For example, when the L in A-L-Z is a single bond, the A-L-Z structure is actually A-Z.

When a substitute is vacant, it means that there is no substitute. For example, when X is vacant in A-X, the A-X structure is actually A.

When a substituent does not specify the atom through which the substituent is bonded to the substitutable group, such substituent may be bonded through any of its atoms. For example, a pyridyl group as a substituent may be linked to the substituting group through any of the carbon atoms of the pyridine ring.

When no link direction is specified for a linking group, its link direction can be any. For example, when the connecting group L in represents -MW-, then -MW- can be linked to cycle A and cycle B in left-to-right reading order to form a combination or it can be connected to cycle A and cycle B in the opposite direction, forming a combination The combination of connecting groups, substituents and/or variants thereof is only permitted if such a combination results in the formation of a stable compound.

Unless otherwise indicated, the term "halogen" by itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.

Unless otherwise indicated, the term "C _1-3 alkoxy group" means an alkyl group containing from 1 to 3 carbon atoms and connected to the rest of the molecule through an oxygen atom. The C _1-3 alkoxy group includes C _1-2 , C _2-3 , C ₃ , C ₂ alkoxy groups and the like. Examples of C _1-3 alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

Unless otherwise indicated, the term "C _1-3 alkyl" is used to refer to a linear or branched saturated hydrocarbon group of 1-3 carbon atoms. The C _1-3 alkyl group includes C _1-2 and C _2-3 alkyl groups, and the like. It can be monovalent (eg methyl), divalent (eg methylene) or multivalent (eg methenyl). Examples of C _1-3 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

Unless otherwise indicated, the terms "5-6 membered heteroaromatic ring" and "5-6 membered heteroaryl" may be used interchangeably. The term "5-6 membered heteroaryl" means a monovalent group containing a conjugated π electron system and consisting of 5 or 6 ring atoms, of which 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the remaining atoms are carbon atoms, where the nitrogen atom is optionally quaternized, and the nitrogen and sulfur atoms are optionally oxidized (for example, NO and S(O) _p , p is 1 or 2). The 5-6 membered heteroaryl group may be attached to the rest of the molecule via a heteroatom or via a carbon atom. A 5-6 membered heteroaryl group includes 5-membered and 6-membered heteroaryl groups. Examples of 5-6 membered heteroaryl groups include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.). p.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl and the like), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl and the like), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl and 4H-1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl and the like), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl and the like), furyl (including 2-furyl and 3- furyl and the like), thienyl (including 2-thienyl and 3-thienyl and the like), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl and the like), pyrazinyl or pyrimidinyl ( including 2-pyrimidinyl and 4-pyrimidinyl and the like).

Unless otherwise indicated, C _n-n+m or C _n -C _n+m includes any number of carbon atoms from n to n+m. For example, C _1-12 includes C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , C ₇ , C ₈ , C ₉ , C ₁₀ , C ₁₁ and C ₁₂ . C _n-n+m or C _n -C _n+m also includes any range from n to n+m. For example, C _1-12 includes C _1-3 , C _1-6 , C _1-9 , C _3-6 , C _3-9 , C _3-12 , C _{6-9 ,} C _6-12 , C _{9- 12} etc. Similarly, a ring from n-membered to n+m-membered means that the number of atoms in the ring is from n to n+m. For example, a 3-12 membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring, a 10-membered ring, 11 -membered ring and 12-membered ring. An n-membered to n+m-membered ring means that the number of atoms in the ring includes any range from n to n+m. For example, a 3-12-membered ring includes a 3-6-membered ring, a 3-9-membered ring, a 5-6-membered ring, a 5-7-membered ring, a 6-7-membered ring, a 6-8-membered ring and a 6 to 10 membered ring, and the like.

The term "leaving group" means a functional group or atom that can be replaced by another functional group or atom during a substitution reaction. For example, representative leaving groups include triflat; chlorine, bromine and iodine; a sulfonate group such as mesylate, tosylate, p-bromobenzenesulfonate, p-toluenesulfonate and the like; an acyloxy group such as acetoxy, trifluoroacetoxy and the like.

The term "protecting group" includes, but is not limited to, "amino protecting group", "hydroxy protecting group", or "thio protecting group". The term "amino protecting group" means a protecting group suitable for blocking side reactions at the nitrogen atom of an amino group. Representative amino protecting groups include (but are not limited to): formyl; acyl such as an alkanoyl (eg acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyl such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl such as benzyl (Bn), trityl (Tr), 1,1-bis-(4'-methoxyphenyl)methyl; silyl such as trimethylsilyl (TMS) and t-butyldimethylsilyl (TBS); and the like. The term "hydroxy protecting group" means a protecting group suitable for blocking side reactions at a hydroxy group. Representative hydroxy protecting groups include (but are not limited to): alkyl such as methyl, ethyl, and tert-butyl; acyl such as alkanoyl (eg acetyl); arylmethyl such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (benzhydryl, DPM); silyl such as trimethylsilyl (TMS) and t-butyldimethylsilyl (TBS); and the like.

The structures of the compounds of the present invention can be confirmed by standard methods known to those skilled in the art. If the present description relates to the absolute configuration of a compound, then the absolute configuration can be confirmed by methods well known in the art, such as single crystal x-ray diffraction analysis. In the method of X-ray diffraction analysis, data on the intensity of diffraction on the grown single crystal are collected using a Bruker D8 diffractometer with a CuKα radiation source in the scanning mode ϕ/ωscan; after collecting this information, the crystal structure is further analyzed by a direct method (Shelxs97) to determine the absolute configuration.

The compounds of the present invention may be prepared by various synthetic methods well known to those skilled in the art, including the particular embodiments described below, variants resulting from the combination of the particular embodiments described below with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. in this area. Alternative embodiments include (but are not limited to) the embodiments described herein.

The solvents used in the present invention are commercially available.

The following abbreviations and abbreviations are used in this text: aq. = water; NaNO ₂ means sodium nitrite; H ₂ SO ₄ means sulfuric acid; CuCl ₂ means copper chloride; LiOH⋅H ₂ O means lithium hydroxide monohydrate; NaBH ₃ CN is sodium cyanoborohydride; HCl/EtOAc means a solution of hydrogen chloride in ethyl acetate; HCl means hydrochloric acid; CO ₂ means carbon dioxide; ACN means acetonitrile; FA stands for formic acid; H ₂ O means water; NH ₃ H ₂ O means ammonia; Na ₂ SO ₄ means sodium sulfate; MgSO ₄ means magnesium sulfate; NCS means N-chlorosuccinimide; NH ₄ HCO ₃ means ammonium bicarbonate; Pd/C means palladium on carbon; psi is a unit of pressure; MeCN means acetonitrile; TsOH⋅H ₂ O is p-toluenesulfonic acid monohydrate; Pd(dppf)Cl ₂ is [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride; Neu means neutral; HPLC stands for High Performance Liquid Chromatography; HATU means O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; K ₂ CO ₃ means potassium carbonate; EDC means N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride; m-CPBA means 3-chloroperbenzoic acid; equiv. means equivalent or equivalence; T ₃ P means propylphosphonic anhydride; CDI means diimidazole carbonyl; DCM means dichloromethane; PE means petroleum ether; DIAD means diisopropyl azodicarboxylate; DMF means N,N-dimethylformamide; DMSO means dimethyl sulfoxide; EtOAc means ethyl acetate; EtOH means ethanol; MeOH means methanol; CBz means benzyloxycarbonyl, which is an amino protecting group; BOC means tert-butoxycarbonyl which is an amino protecting group; HOAc means acetic acid; NaCNBH ₃ is sodium cyanoborohydride; rt or rt means room temperature; O/N = during the night; THF means tetrahydrofuran; Boc ₂ O means di-tert-butyl dicarbonate; TFA means trifluoroacetic acid; SOCl ₂ means thionyl chloride; CS ₂ means carbon disulfide; TsOH means p-toluenesulfonic acid; NFSI means N-fluoro-N-(phenylsulfonyl)benzenesulfonamide; n-Bu ₄ NF means tetrabutylammonium fluoride; iPrOH means 2-propanol; m.p. means melting point; LDA means lithium diisopropylamide; IPAm means isopropylamine; DIPEA means N,N-diisopropylethylamine; Xantphos is 4,5-bis(diphenylphosphino)-9,9-dimethylxanthane; t-Bu Xphos is 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl; DPPP means 1,3-bis(diphenylphosphino)propane; Et ₃ SiH means triethylsilane; DMAP means 4-dimethylaminopyridine; Py means pyridine; DPBS stands for Dulbecco's Phosphate Buffer; TNF-α means tumor necrosis factor; IC ₅₀ means the concentration of half-maximal inhibition; mg means milligrams, kg means kilograms; and HEPES is N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid buffer.

Compounds are named according to general naming conventions or by using the ChemDraw® program, and commercially available compounds are named according to names from their suppliers.

Detailed description of the invention

The present invention is described in more detail below with the help of examples. However, the examples given do not limit the scope of the present invention in any way. The present invention is described in detail in the present text, and embodiments thereof are also described. It will be apparent to those skilled in the art that various changes and modifications can be made to the embodiments described herein without departing from the spirit and scope of the present invention.

Comparative Example 1: Fragment BB-1

Synthesis path:

Step 1: Synthesis of Compound BB-1-3

BB-1-1 (2.60 g, 17.31 mmol, 1.00 equiv.) and EtOH solvent (11.25 ml) and isopropyl ether (37.50 ml) were placed in a pre-dried 100 ml flask. BB-1-2 (2.59 g, 17.83 mmol, 1.03 eq.) was slowly added with stirring, and the reaction solution was stirred at 20°C for 12 hours. The reaction solution was filtered, and the filter cake was washed with isopropyl ether (30 ml*3) to give compound BB-1-3. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 9.95 (s, 1H), 9.90 (s, 1H), 7.39 - 7.08 (m, 6H), 4.21 - 4.19 (m, 2H), 3.82 (s, 2H), 1.27 - 1.23 (m, 3H); LCMS m/z = 250.1 [M+H] ⁺ .

Step 2: Synthesis of Compound BB-1-4

Compound BB-1-3 (7.00 g, 28.08 mmol, 1.00 eq.) and xylene (100.00 ml) were placed in a pre-dried 250 ml flask. Installed the Dean-Stark nozzle. The reaction mixture was heated at 180°C for 10 hours. 100 ml of isopropyl ether was added to the reaction system and the mixture was stirred for 30 minutes in an ice bath. The reaction mixture was filtered and the filter cake was washed with 50 ml of n-hexane to give compound BB-1-4. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 14.44 (broad s, 1H), 7.42 - 7.14 (m, 4H), 4.29 (q, J = 6.5 Hz, 2H), 4.13 (broad s, 2H), 1.28 (broad t, J = 7.0 Hz, 3H); LCMS m/z = 232.2 [M+H] ⁺ .

Step 3: Synthesis of Compound BB-1

Compound BB-1-4 (4.10 g, 17.73 mmol, 1.00 equiv.), tetrahydrofuran (1.00 ml) and water (200.00 μl) were placed in a flask, then lithium hydroxide monohydrate (1.74 g, 41.49 mmol, 2.34 equiv.) was added. The solution was stirred at 20°C for 12 hours. The solvent was removed under reduced pressure. Added dropwise 25 ml of 1N. aqueous hydrochloric acid solution with stirring until pH 2. Compound BB-1 was obtained after suction filtration. LCMS m/z = 202.1 [MH] ^- .

Comparative Example 2: Fragment BB-2

Synthesis path:

Step 1: Synthesis of Compound BB-2-2

To a solution of BB-2-1 (2.5 g, 15.51 mmol, 2.94 ml, 1 equiv.) in dichloromethane (40 ml) was added methanol (4.97 g, 155.13 mmol, 6.28 ml, 10 equiv.) at 23°C. Then TMSCHN ₂ (2 M, 9.31 ml, 1.2 eq.) was added and the mixture was stirred for another 2 hours. The starting compounds disappeared completely according to TLC monitoring (petroleum ether:ethyl acetate = 5:1). 0.1 ml of glacial acetic acid was added to the reaction solution, and the solution was evaporated under reduced pressure to obtain BB-2-2.

Step 2: Synthesis of Compound BB-2-3

To a solution of BB-2-2 (3 g, 17.12 mmol, 1 eq.) in methanol (30 ml) was added hydrazine hydrate (3.21 g, 51.37 mmol, 3.12 ml, 80% pure, 3 eq.) and the mixture was stirred for oil bath at 70°C for 2 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was evaporated under reduced pressure to remove the solvent, and 50 ml of ethyl acetate was added. The mixture was washed with 30 ml of a saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was evaporated under reduced pressure to give BB-2-3. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 9.25 (broad s, 1 H), 7.66 - 7.74 (m, 2 H), 7.45 - 7.65 (m, 2 H), 4.24 (s, 2 H ), 3.43 (s, 2 H); LCMS m/z = 176.1 [M+H] ⁺ .

Step 3: Synthesis of Compound BB-2-4

BB-1-2 (2.07 g, 14.27 mmol, 1 equiv.), ethanol (10 ml) and isopropyl ether (20 ml) were placed in a pre-dried three-necked flask, then BB-2-3 (2.5 g, 14.27 mmol, 1 equivalent). The mixture was allowed to react at 25° C. for 5 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was filtered, and the filter cake was washed with 50 ml of petroleum ether to obtain BB-2-4. LCMS m/z = 275.2 [M+H] ⁺ .

Step 4: Synthesis of Compound BB-2-5

BB-2-4 (0.5 g, 1.82 mmol, 1 equiv.), molecular sieves (4 Å, 0.05 g), and xylene (15 mL) were placed in a vial for reactions in a microwave oven. The microwave reaction vial was sealed and the reaction mixture was stirred in a microwave reactor at 150° C. for 4 hours. Monitoring by LCMS showed the appearance of the signal of the target product. 30 ml of acetonitrile was added to the reaction solution, and the mixture was filtered. The filtrate was evaporated under reduced pressure to give BB-2-5. LCMS m/z = 257.1 [M+H] ⁺ .

Step 5: Synthesis of Compound BB-2

To a solution of BB-2-5 (1 g, 3.90 mmol, 1 equiv.) in tetrahydrofuran (50 ml) was added a solution of lithium hydroxide monohydrate (654.96 mg, 15.61 mmol, 4 equiv.) in water (15 ml) at 21°C and the mixture was stirred for 12 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. 15 ml of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 ml). The resulting aqueous phase was brought to a pH value of about 3 by adding 2N. aqueous hydrochloric acid solution and lyophilized to give BB-2. LCMS m/z = 227.0 [MH] ^- .

Comparative Example 3: Fragment BB-3

Synthesis path:

Step 1: Synthesis of Compound BB-3-3

BB-3-2 (20.00 g, 97.46 mmol, 1.00 equiv.) was dissolved in DMF (50 ml), and the resulting solution was added dropwise to a suspension of NaH (8.19 g, 204.67 mmol, 60% purity, 2.10 equiv.) in DMF (150 ml). Compound BB-3-1 (13.75 g, 97.46 mmol, 10.26 ml, 1.00 eq.) was then added dropwise. The reaction solution was stirred at 20° C. for 16 hours. Then, ethyl acetate (200 ml) and water (200 ml) were added to the reaction solution, and the aqueous phase was adjusted to a pH of about 1 by adding hydrochloric acid. The mixture was extracted with ethyl acetate (200 ml). The organic phase was dried over anhydrous magnesium sulfate and filtered, the filtrate was evaporated under reduced pressure to give compound BB-3-3. LCMS m/z = 227.1 [M-100+H] ⁺ .

Step 2: Synthesis of Compound BB-3-4

A solution of compound BB-3-3 (20.00 g, 61.29 mmol, 1.00 equiv.) and wet Pd/C (5.00 g, 5% pure) in methanol (200.00 ml) was stirred at 20°C under a hydrogen atmosphere (15 psi . inch) 5 hours. After completion of the reaction, the solution was filtered, and the filtrate was evaporated under reduced pressure to give compound BB-3-4. LCMS m/z = 297.1 [M+H] ⁺ .

Step 3: Synthesis of Compound BB-3-5

HATU (12.83 g , 33.75 mmol, 1.00 equiv.). The reaction mixture was stirred at 20°C for 2 hours. After completion of the reaction, water (200 ml) and ethyl acetate (200 ml) were added to the solution, and the layers were separated. The organic phase was evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1~1:1) to obtain compound BB-3-5. LCMS m/z = 223.1 [M-56+H] ⁺ .

Step 4: Synthesis of Compound BB-3-6

Compound BB-3-5 (6.00 g, 21.56 mmol, 1.00 eq.), DMF (60.00 ml) and cesium carbonate (10.54 g, 32.34 mmol, 1.50 eq.) were placed in a dried 250 ml three-necked flask, then iodomethane ( 3.06 g, 21.56 mmol, 1.34 ml, 1.00 eq.) drop by drop. Controlled the temperature in the reaction mixture so that it did not rise above 20°C. The reaction mixture was stirred at 16°C for 16 hours. After completion of the reaction, water (100 ml) and ethyl acetate (100 ml) were added and the layers were separated. The organic phase was washed with saturated aqueous sodium chloride solution (100 ml * 2), dried over magnesium sulfate and filtered. The filtrate was evaporated under reduced pressure to give compound BB-3-6. LCMS m/z = 237.1 [M-56+H] ⁺ .

Step 5: Synthesis of Compound BB-3-7

Sodium carbonate (362.57 mg, 3.42 mmol, 5 equiv.) was added to a solution of phosphorus pentasulfide (760.35 mg, 3.42 mmol, 363.80 μL, 5 equiv.) in THF (10 mL) at 20°C, and the mixture was stirred for another 0.5 h. Then BB-3-6 (0.2 g, 684.16 µmol, 1 eq.) was added and the reaction solution was heated in an oil bath at 70°C with stirring for 9.5 hours. Monitoring by LCMS showed the appearance of the signal of the target product. 20 ml of ethyl acetate was added to the reaction solution, and the mixture was filtered. The resulting filtrate was evaporated under reduced pressure to give compound BB-3-7. LCMS m/z = 253.1 [M-56+H] ⁺ .

Step 6: Synthesis of Compound BB-3

To BB-3-7 (630.84 mg, 2.05 mmol, 1 eq.) was added a solution of hydrogen chloride in ethyl acetate (4M, 20.00 ml, 39.11 eq.) and the mixture was stirred at 23°C for 1 hour. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was evaporated under reduced pressure to give a crude product, then 30 ml of a saturated sodium bicarbonate aqueous solution was added. The mixture was extracted with ethyl acetate (15 ml * 3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give compound BB-3. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.21 - 7.32 (m, 3 H), 7.15 - 7.19 (m, 1 H), 4.40 (dd, J = 9.92, 6.62 Hz, 1 H), 4.10 (dd , J = 11.36, 9.81 Hz, 1 H), 3.77–3.94 (m, 4 H); LCMS m/z = 209.1 [M+H] ⁺ .

Comparative Example 4: Fragment BB-4

Synthesis path:

Step 1: Synthesis of Compound BB-4-2

To a solution of BB-3-2 (13.13 g, 63.98 mmol, 1 equiv.) in DMF (100 ml) was added NaH (5.12 g, 127.96 mmol, 60% pure, 2 equiv.) at 0°C, and the mixture was stirred 1 hour. Then BB-4-1 (10 g, 70.38 mmol, 1.1 eq.) was added and the mixture was stirred at 28°C for 9 hours. Monitoring by LCMS showed the appearance of the signal of the target product. The reaction solution was added to 1000 ml of water, and the mixture was extracted with ethyl acetate (300 ml*4). The aqueous phase was brought to a pH value of about 5 by adding 2N. hydrochloric acid solution and then extracted with ethyl acetate (300 ml * 5). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give compound BB-4-2. LCMS m/z = 326.0 [MH] ^- .

Step 2: Synthesis of Compound BB-4-3

Wet Pd/C (10 g, 30.55 mmol, 5% purity) was added to a solution of BB-4-2 (20.00 g, 61.11 mmol, 1 equiv.) in methanol (200 mL) at 23°C. The mixture was stirred under a hydrogen atmosphere (15 psi) for 10 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was filtered, and the resulting filtrate was evaporated under reduced pressure to give compound BB-4-3. LCMS m/z = 298.1 [M+H] ⁺ .

Step 3: Synthesis of Compound BB-4-4

To a solution of BB-4-3 (16.00 g, 53.82 mmol, 1 eq.) in ethyl acetate (400 ml) was added N,N-diisopropylethylamine (20.87 g, 161.45 mmol, 28.12 ml, 3 eq.) and 2,4,6 -tributyl-1,3,5,2,4,6-trioxatrifosfinan-2,4,6-trioxide (68.49 g, 107.63 mmol, 64.01 ml, 50% solution in ethyl acetate, 2 eq.) at 23°C . The mixture was stirred for another 2 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was washed with a saturated sodium chloride aqueous solution (200 ml*2). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1~1:1) to obtain compound BB-4-4. LCMS m/z = 224.1 [M-56+H] ⁺ .

Step 4: Synthesis of Compound BB-4-5

Cesium carbonate (1.40 g, 4.30 mmol, 1.2 equiv.) and iodomethane (609.86 mg, 4.30 mmol, 267.48 µl, 1.2 equiv.) at 20°C. The mixture was stirred for 1 hour. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was poured into 100 ml of water, and the mixture was extracted with ethyl acetate (30 ml*5). The organic phases were combined and washed with 50 ml of a saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was evaporated under reduced pressure to give compound BB-4-5. LCMS m/z = 238.1 [M-56+H] ⁺ .

Step 5: Synthesis of Compound BB-4-6

To a solution of phosphorus pentasulfide (3.79 g, 17.05 mmol, 10 equiv.) in THF (30 ml) was added sodium carbonate (1.81 g, 17.05 mmol, 10 equiv.) at 20°C, and the mixture was stirred for 0.5 h. Then BB-4-5 (0.5 g, 1.70 mmol, 1 eq.) was added and the reaction solution was heated in an oil bath at 70° C. with stirring for 9.5 hours. According to LCMS monitoring, the starting compounds almost completely disappeared, and the signal of the target product was observed. The reaction solution was filtered to obtain a filtrate. 50 ml of ethyl acetate was added to the filtrate, and the mixture was washed with 30 ml of a saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1~1:1) to obtain compound BB-4-6. LCMS m/z = 310.3 [M+H] ⁺ .

Step 6: Synthesis of compound BB-4.

To BB-4-6 (0.6 g, 1.94 mmol, 1 eq.) was added a solution of hydrogen chloride in ethyl acetate (4 M, 20 ml, 41.25 eq.) and the mixture was stirred at 23°C for 1 hour. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was evaporated under reduced pressure to give a crude product, and 30 ml of a saturated sodium bicarbonate aqueous solution was added. The mixture was extracted with ethyl acetate (20 ml * 3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give compound BB-4. LCMS m/z = 210.1 [M+H] ⁺ .

Comparative Example 5: Fragment BB-5

Synthesis path:

Step 1: Synthesis of Compound BB-5-1

Deuterated iodomethane (101.64 mg, 716.10 µmol, 43.62 µl, 2 equiv.) and cesium carbonate (349.98 mg, 1.07 mmol , 3 eq.) at 25°C, and the mixture was stirred for another 2 hours. Ethyl acetate (20 ml) was added to the reaction solution, then the mixture was washed with a saturated aqueous sodium chloride solution (10 ml) and filtered. The filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by suspension in methyl tert-butyl ether (5 ml) to give compound BB-5-1. LCMS m/z = 297.2 [M+H] ⁺ .

Step 2: Synthesis of Compound BB-5-2

To a solution of phosphorus pentasulfide (2.25 g, 10.12 mmol, 1.08 ml, 10 equiv.) in THF (10 ml) was added sodium carbonate (1.07 g, 10.12 mmol, 10 equiv.) at 30°C, and the mixture was stirred for 0.5 h. Then BB-5-1 (0.3 g, 1.01 mmol, 1 eq.) was added and the reaction solution was stirred in an oil bath at 70°C for 10 hours. Ethyl acetate (20 ml) was added to the reaction solution. The mixture was filtered through celite and the filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate=10:1~1:1) to give compound BB-5-2. LCMS m/z = 312.9 [M+H] ⁺ .

Step 3: Synthesis of Compound BB-5

HCl/EtOAc (4M, 5.69 ml, 59.27 eq) was added to BB-5-2 (0.12 g, 384.12 µmol, 1 eq) at 28°C and the mixture was stirred for 1 hour. The reaction solution was evaporated under reduced pressure to give compound BB-5. LCMS m/z = 213.1 [M+H] ⁺ .

Comparative Example 6: Fragment BB-6

Synthesis path:

Step 1: Synthesis of Compound BB-6-2

Benzyl bromide (2.44 g, _14.27 mmol _, 1.70 ml, 1 eq.) at 25°C, and the mixture was stirred for 3 hours. According to LCMS monitoring, the original compounds completely reacted, and a signal appeared with the target mass. The reaction solution was poured into water (100 ml), and the mixture was extracted with ethyl acetate (30 ml*5). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was evaporated under reduced pressure. The crude product was purified by silica gel flash column chromatography (petroleum ether:ethyl acetate=10:1~1:1) to obtain BB-6-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.58 (d, J = 1.00 Hz, 1 H), 7.55 (s, 1 H), 7.33 - 7.40 (m, 3 H), 7.14 - 7.20 (m, 2 H), 5.13 (s, 2 H), 4.34 (q, J = 7.20 Hz, 2 H), 1.36 (t, J = 7.20 Hz, 3 H); LCMS m/z = 231.1 [M+1] ⁺ .

Step 2: Synthesis of Compound BB-6

To a solution of BB-6-2 (1.50 g, 6.51 mmol, 1 equiv.) in tetrahydrofuran (26 ml) was added a solution of lithium hydroxide monohydrate (2.19 g, 52.11 mmol, 8 equiv.) in water (26 ml) at 28°C and the mixture was stirred for another 12 hours. According to LCMS monitoring, the original compounds completely reacted, and a signal appeared with the target mass. The reaction solution was evaporated under reduced pressure. After removal of the solvent, water (20 ml) was added and the mixture was adjusted to a pH of about 2 by adding 2N. hydrochloric acid solution. The precipitate that formed was filtered off to give BB-6. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 7.87 (d, J = 2.20 Hz, 2 H), 7.27 - 7.43 (m, 5 H), 5.24 (s, 2 H); LCMS m/z = 201.1 [M-1] ^- .

Comparative Example 7: Fragment BB-7

Synthesis path:

Step 1: Synthesis of Compound BB-7-2

Diethyl oxalate (4.77 g, 32.62 mmol, 4.45 ml, 1.1 equiv.) and sodium ethoxide (2.62 g, 38.55 mmol, 1.3 eq.) at 0°C, and the mixture was heated at 20°C for 12 hours. According to LCMS analysis, the starting compounds were completely reacted, and the signal of the target product was observed. The reaction solution was evaporated under reduced pressure, the residue was diluted with ethyl acetate (200 ml) and washed with saturated aqueous sodium chloride solution (200 ml). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was evaporated under reduced pressure to give a crude product which was directly used in the next step without purification. Received connection BB-7-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 6.87 - 7.19 (m, 4 H), 5.78 (br. s, 1 H), 4.04 (br. s, 2 H), 2.92 - 3.44 (m, 2 H ), 1.18 (broad s, 3 H); LCMS m/z = 269.1[M+1] ⁺ .

Step 2: Synthesis of Compound BB-7-3

To a solution of BB-7-2 (7 g, 26.05 mmol, 1 eq.) in ethanol (200 ml) was added hydroxylamine hydrochloride (1.81 g, 26.05 mmol, 1 eq.) and the mixture was heated at 80°C for 12 hours . According to LCMS analysis, the starting compounds were completely reacted, and the signal of the target product was observed. The reaction solution was evaporated under reduced pressure to give BB-7-3. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.33 (m, 2 H); 7.20 (m, 2 H), 6.35 (s, 1 H), 4.41 - 4.47 (m, 2 H), 4.13 (q, J = 7.20 Hz, 2 H), 1.30 - 1.33 (t, 3 H); LCMS m/z = 266.1[M+1] ⁺ .

Step 3: Synthesis of Compound BB-7-4

Pd/C (4 g, 5% purity) was added to a solution of BB-7-3 (4 g, 15.06 mmol, 1 eq.) in ethanol (100 ml) and the mixture was heated at 30° C. under hydrogen (30 lb. /square inch) 12 hours. According to LCMS analysis, the starting compounds were completely reacted, and the signal of the target product was observed. The reaction solution was filtered through Celite, and the filtrate was evaporated under reduced pressure to give a crude product. The crude product was purified by silica gel column chromatography (dichloromethane:methanol=10:1) to give compound BB-7-4. LCMS m/z = 232.1 [M+1] ⁺ .

Step 4: Synthesis of Compound BB-7

To a solution of BB-7-4 (0.5 g, 2.16 mmol, 1 equiv.) in tetrahydrofuran (15 ml) and water (5 ml) was added lithium hydroxide monohydrate (453.17 mg, 10.80 mmol, 5 equiv.), and the mixture was heated at 30°C for 2 hours. The starting compounds reacted completely and a new spot was observed on TLC (petroleum ether:ethyl acetate = 5:1). The reaction solution was evaporated under reduced pressure to remove the organic solvent, and a 2M hydrochloric acid solution was added to adjust the pH to about 2. A white precipitate precipitated. The mixture was filtered and the filter cake was collected to give BB-7. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 7.24 - 7.41 (m, 5 H), 6.57 (s, 1 H), 4.22 (s, 2 H).

Comparative Example 8: Fragment BB-8

Synthesis path:

Step 1: Synthesis of Compound BB-8-2

Cesium carbonate (6.67 g, 20.46 mmol, 1.3 equiv.) and benzyl bromide (3.23 g, 18.88 mmol, 2.24 ml, 1.2 eq.), and the mixture was stirred at 20°C for 2 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. Water (30 ml) and ethyl acetate (30 ml) were added to the reaction solution, and the layers were separated. The organic phase was washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel flash column chromatography (petroleum ether:ethyl acetate=10:1-3:1) to give BB-8-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.11 (s, 1 H), 7.34 - 7.43 (m, 3 H), 7.27 - 7.33 (m, 2 H), 5.43 (s, 2 H), 4.00 ( s, 3H); LCMS m/z = 218.1 [M+1] ⁺ .

Step 2: Synthesis of Compound BB-8

BB-8-2 (0.3 g, 1.38 mmol, 1 eq.) was dissolved in tetrahydrofuran (12 ml) and water (4 ml) and lithium hydroxide monohydrate (173.85 mg, 4.14 mmol, 3 eq.) was added. The reaction mixture was stirred at 25°C for 12 hours. According to LCMS monitoring, the original compounds completely reacted, and a peak of the target product appeared. Ethyl acetate (10 ml) was added to the reaction solution, and the aqueous phase was adjusted to pH 3-4 by adding 1M hydrochloric acid. There was a precipitate. The mixture was filtered to give BB-8. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ =13.25 (broad s, 1 H), 8.78 (s, 1 H), 7.27 - 7.40 (m, 5 H), 5.47 (s, 2 H); LCMS m/z = 204.1 [M+1] ⁺ .

Comparative Example 9: Fragment BB-9

Synthesis path:

Step 1: Synthesis of Compound BB-9-2

NaNO ₂ (662.86 mg, 9.60 mmol, 1.5 equiv.) was added to a solution of BB-9-1 (1 g, 6.40 mmol, 1 equiv.) in H ₂ SO ₄ (1 M, 12.80 ml, 2 equiv.) and water (10 ml) at 0°C, and the mixture was stirred at 0°C for 0.5 hours. CuCl ₂ (1.72 g, 12.80 mmol, 2 equiv.) was dissolved in concentrated hydrochloric acid (15 ml) and the resulting solution was added to the above mixture. The resulting mixture was stirred at 0°C for 1.5 hours. According to LCMS monitoring, the original compounds completely reacted, and a peak of the target product appeared. Ethyl acetate (10 ml) was added to the reaction solution, and the aqueous phase was extracted with ethyl acetate (15 ml*2). The organic phase was washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure. The crude product was purified by silica gel flash column chromatography (petroleum ether:ethyl acetate=10:1~1:2) to obtain BB-9-2. ¹ H NMR (400 MHz, CDCl ₃ ) = 4.46 (q, J = 21.60 Hz, 2 H), 1.38 (t, J = 14.40 Hz, 3 H); LCMS m/z = 176.0 [M+1] ⁺ .

Step 2: Synthesis of Compound BB-9-3

Cesium carbonate (1.84 g, 5.63 mmol, 1.3 equiv.) and 3-cyanobenzyl bromide (889.31 mg, 5.20 mmol, 617.58 μl, 1.2 eq.), and the reaction mixture was stirred at 20°C for 2 hours. According to LCMS monitoring, the original compounds completely reacted, and a peak of the target product appeared. Water (10 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 ml*2). The organic phase was washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure, and the crude product was purified by silica gel flash column chromatography (petroleum ether:ethyl acetate=20:1~2:1) to give BB-9-3. LCMS m/z = 291.0 [M+1] ⁺ .

Step 3: Synthesis of Compound BB-9

BB-9-3 (0.08 g, 301.10 µmol, 1 eq.) was dissolved in tetrahydrofuran (6 ml) and water (2 ml), and lithium hydroxide monohydrate (37.90 mg, 903.29 µmol, 3 eq.) was added. The mixture was heated at 2°C for 12 hours. According to LCMS monitoring, the original compounds completely reacted, and a peak of the target product appeared. Ethyl acetate (5 ml) was added to the reaction solution. The aqueous phase was adjusted to a pH of about 6 with 1M hydrochloric acid and lyophilized to give BB-9. LCMS m/z = 263.1 [M+1] ⁺ .

Comparative Example 10: Fragment BB-10

Synthesis path:

Step 1: Synthesis of Compound BB-10-1

To a solution of BB-1-1 (3 g, 19.98 mmol, 1 equiv.) in dichloromethane (90 ml) was added triethylamine (6.06 g, 59.93 mmol, 8.34 ml, 3 equiv.) 2.91 ml, 1.3 eq.), and the mixture was stirred at 25°C for 5 hours. According to LCMS analysis, the starting compounds were completely reacted, and the signal of the target product was observed. The reaction was quenched by adding water (60 ml) and extracted with dichloromethane (60 ml*3). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was evaporated under reduced pressure. The crude product was purified by silica gel flash column chromatography (petroleum ether:ethyl acetate=5:1-1:1) to give BB-10-1. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 10.73 (br. s, 1 H), 10.30 (br. s, 1 H), 7.22 - 7.34 (m, 5 H), 4.26 (q.v., J = 7.00 Hz, 2 H), 3.51 (s, 2 H), 1.27 (t, J = 7.10 Hz, 3 H); LCMS m/z = 251.1[M+1] ⁺ .

Step 2: Synthesis of Compound BB-10

To a solution of BB-10-1 (529.69 mg, 2.12 mmol, 1 equiv.) in acetonitrile (15 ml) was added phosphorus oxychloride (486.82 mg, 3.17 mmol, 295.04 µl, 1.5 equiv.). The reaction mixture was stirred at 90°C for 2 hours. The reaction solution was poured into a saturated sodium bicarbonate solution (20 ml) and the mixture was vigorously stirred for 5 minutes. The mixture was extracted twice with ethyl acetate (20 ml). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give BB-10. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.23 - 7.10 (m, 5H), 4.36 - 4.29 (m, 2H), 4.12 (s, 2H), 1.34 - 1.23 (m, 3H); LCMS m/z = 233.1 [M+1] ⁺ .

Comparative Example 11: Detail of BB-11A, BB-11B

Synthesis path:

Step 1: Synthesis of Compound BB-11-2

To a solution of BB-11-1 (5 g, 26.01 mmol, 1 equiv.) in glacial acetic acid (39 ml) and tetrahydrofuran (150 ml) was added tert-butylcarbazate (17.19 g, 130.07 mmol, 5 equiv.), and the mixture heated at 55°C for 12 hours. The mixture was cooled to room temperature (25°C) and NaBH ₃ CN (8.17 g, 130.07 mmol, 5 eq.) was added in portions. The mixture was heated to 55°C and left to pass the reaction for 12 hours. The reaction solution was cooled to room temperature, and a saturated sodium bicarbonate solution was added to adjust the pH to 7~8. The mixture was extracted with ethyl acetate (150 ml*2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give BB-11-2. LCMS m/z = 221.1 [M-56+1] ⁺ .

Step 2: Synthesis of Compound BB-11-3

BB-11-2 (7 g, 25.33 mmol, 1 eq.) was added to a solution of HCl/EtOAc (4M, 60 ml) and the resulting mixture was heated at 25°C for 2 hours. The reaction solution was evaporated under reduced pressure to give a crude product. A saturated sodium bicarbonate solution was added to the crude product, adjusting the pH to 7~8. The mixture was extracted with ethyl acetate (50 ml*2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give crude product BB-11-3. LCMS m/z = 177.1 [M+1] ⁺ .

Step 3: Synthesis of Compound BB-11-4.

To a solution of BB-11-3 (4 g, 22.70 mmol, 1 eq.) in ethanol (50 ml) was added ethyl 2-ethoxy-2-iminoacetate (16.47 g, 113.50 mmol, 5 eq.) and the mixture was heated at 85 °C for 12 hours. The reaction solution was evaporated under reduced pressure to give a crude product. The crude product was purified by column chromatography (dichloromethane:methanol=10:0 to 5:1) to give BB-11-4. LCMS m/z = 276.2 [M+1] ⁺ .

Step 4: Synthesis of Compound BB-11-5

BB-11-4 (1.3 g, 4.72 mmol, 1 eq.) was added to phosphorus oxychloride (30 ml) at 25°C and the mixture was heated at 120°C for 3 hours. The reaction solution was cooled to room temperature and then slowly poured into warm water (150 ml). The mixture was adjusted to pH 7~8 with saturated aqueous sodium bicarbonate and then extracted with ethyl acetate (100 ml*2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by column chromatography (dichloromethane:methanol=10:0 to 10:1) to give BB-11-5. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 7.34 - 7.43 (m, 3 H), 7.21 - 7.28 (m, 2 H), 5.58 (broad t, J = 7.12 Hz, 1 H), 4.27 (broad dd, J = 6.80, 5.04 Hz, 2 H), 2.88–3.28 (m, 4 H), 1.26 (broad t, J = 7.12 Hz, 3 H). LCMS m/z = 258.2 [M+1] ⁺ .

Step 5: Synthesis of Compound BB-11A and BB-11B

Compound BB-11 was analyzed by supercritical liquid chromatography (Chiralcel AD-3 3 µm, 0.46 cm id x 15 ml; Mobile phase: A = SFC CO ₂ and B = MeOH (0.05% IPAm); Gradient: B in A from 10% to 40 % in 6 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm), and it was shown that the retention times of BB-11A and BB-11B were 2.217 min and 2.427 min, respectively. BB-11A and BB-11B were obtained by separation by supercritical liquid chromatography (Column: DAICEL CHIRALPAK AD (250mm*50mm, 10 µm); Mobile phase: [0.1% NH ₃ H ₂ O MEOH]; MeOH%: 30%-30 %, 2.56 min). Compound BB-11A was analyzed by supercritical liquid chromatography (Chiralcel AD-3 3 µm, 0.46 cm id × 15 ml; Mobile phase: A = supercritical carbon dioxide, B = methanol (containing 0.05% IPAm); Gradient: B from 10% to 40 % in 6 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm), and it was shown that the retention time of BB-11A is 2.235 min, and the retention time of BB-11B is 2.439 min.

Comparative Example 12: Fragment BB-12

Synthesis path:

Step 1: Synthesis of compound BB-12.

I ₂ (11.40 g, 44.92 mmol, 9.05 ml, 1 equiv.) and Ag ₂ SO ₄ (14.01 g, 44.92 mmol, 7.61 ml, 1 equiv.) were added to a solution of BB-12-1 in ethanol (100 ml) at 25°C, and the mixture was stirred for 12 hours. The reaction solution was filtered through Celite. Water (150 ml) and ethyl acetate (150 ml) were added to the filtrate and the layers were separated. The aqueous phase was extracted with ethyl acetate (50 ml*2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give the crude product. The crude product was dissolved in ethyl acetate and a solution of hydrogen chloride/ethyl acetate (4 M) was added. The mixture was stirred at room temperature for 0.5 hour and filtered. The precipitate on the filter was separated. Ethyl acetate (50 ml) was added to the precipitate and saturated sodium bicarbonate solution was added to adjust the pH to 8~9. The layers were separated. The aqueous phase was extracted with ethyl acetate (25 ml*2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure (water jet pump, 45°C) to obtain a crude product, and the resulting crude product was purified by column chromatography (petroleum ether:ethyl acetate = 10:1~1:1) to obtain BB-12. ^1H NMR (400 MHz, _CDCl3 ) δ = 7.22 (dt, J = 7.68, 2.37 Hz, 1 H), 6.83 (ddd, J = 10.85, 8.22, 2.76 Hz, 1 H), 3.97 (broad s, 2H).

Comparative Example 13: Fragment BB-13

Synthesis path:

Step 1: Synthesis of Compound BB-13-1

Cesium carbonate (1.84 g, 5.63 mmol, 1.3 equiv.) and benzyl bromide (889.31 mg, 5.20 mmol, 617.58 µl, 1.2 eq.), and the reaction mixture was stirred at 20°C for 2 hours. According to LCMS monitoring, the original compounds completely reacted, and a peak of the target product appeared. Water (10 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 ml*2). The organic phase was washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure. The crude product was purified by silica gel flash column chromatography (petroleum ether:ethyl acetate = 20:1~2:1) to obtain BB-13-1. ¹ H NMR (400 MHz, CDCl ₃ ) δ =7.34 - 7.41 (m, 3 H), 7.29 - 7.34 (m, 2 H), 5.42 (s, 2 H), 4.47 (q, J = 22.40 Hz, 2 H), 1.43 (t, J = 15.60 Hz, 3 H); LCMS m/z = 266.0 [M+1] ⁺ .

Step 2: Synthesis of Compound BB-13

BB-13-1 (0.08 g, 301.10 µmol, 1 eq.) was dissolved in tetrahydrofuran (6 ml) and water (2 ml), and lithium hydroxide monohydrate (37.90 mg, 903.29 µmol, 3 eq.) was added. The mixture was heated at 25°C for 12 hours. According to LCMS monitoring, the original compounds completely reacted, and a peak of the target product appeared. 5 ml of ethyl acetate was added to the reaction solution. The aqueous phase was adjusted to a pH of about 6 with 1M hydrochloric acid and lyophilized to give BB-13. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 7.32 - 7.42 (m, 3 H), 7.25 - 7.30 (m, 2 H), 5.48 (s, 2 H); LCMS m/z = 238.1 [M+1] ⁺ .

Example 1. WX001

Synthesis path:

Step 1: Synthesis of Compound WX001

Compound BB-3 (0.58 g, 2.78 mmol, 1 eq.), Compound BB-1 (849 mg, 4.18 mmol, 1.5 eq.), N,N-diisopropylethylamine (1.26 g, 9.75 mmol, 1.70 ml, 3.5 eq. ) was dissolved in ethyl acetate (10 ml) and then 2,4,6-tributyl-1,3,5,2,4,6-trioxatrifosfinan-2,4,6-trioxide (4.43 g, 6.96 mmol, 4.14 ml, 50% solution in ethyl acetate, 2.5 eq.). After the addition was completed, the mixture was stirred at 25° C. for 12 hours. 20 ml of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (5 ml*2). The organic phases were combined and washed with saturated aqueous sodium chloride solution (10 ml). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography and then by supercritical liquid chromatography (Chiralcel OJ-3, 3 µm, 0.46 cm id × 5 cm L; Mobile phase: A = supercritical carbon dioxide, B = methanol (containing 0.05% IPAm); gradient : B 10% to 40% in 5 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm) to give compound WX001. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.71 (d, J = 7.53 Hz, 1H), 7.20-7.38 (m, 9H), 5.15 - 5.25 (m, 1H), 4.67 (dd, J = 6.53, 9.54 Hz, 1H), 4.29 (dd, J = 9.79, 10.79 Hz, 1H), 4.21 (s, 2H), 3.89 (s, 3H); LCMS m/z = 394.1 [M+H] ⁺ . According to SLC analysis (Chiral column: Column: Chiralpak OD -3, 3 µm, 0.46 cm id × 5 cm L; Mobile phase: A: CO ₂ , B: MeOH (0.05% IPAm); Gradient: B/A = 10% - 40% for 3 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm), ee% was 96.36%.

Example 2. WX002

Synthesis path:

Step 1: Synthesis of Compound WX002

2,4,6-tributyl-1,3, 5,2,4,6-trioxatrifosfinan-2,4,6-trioxide (458.30 mg, 720.19 µmol, 428.32 µl, 50% purity in ethyl acetate, 2.5 eq) and N,N-diisopropylethylamine (111.69 mg, 864.22 µmol, 150.53 μl, 3 eq.), and the mixture was stirred at 25°C for 10 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. 20 ml of ethyl acetate was added to the reaction solution, and the mixture was washed with 15 ml of a saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Xtimate C18 150 mm*25 mm*5 µm; Mobile phase: [water (10 mM NH ₄ HCO ₃ )-ACN]; acetonitrile%: 35%-65%, 10.5 min), receiving connection WX002. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.60 (broad.d, J = 8.16 Hz, 1 H), 7.69 - 7.81 (m, 2 H), 7.51 - 7.65 (m, 3 H), 7.36 - 7.46 (m, 2 H), 7.25 - 7.33 (m, 1 H), 4.97 (dt, J = 10.58, 7.72 Hz, 1 H), 4.35 - 4.52 (m, 2 H), 4.21 (s, 2 H ), 3.79 (s, 3H); LCMS m/z = 419.1 [M+H] ⁺ . According to SLC analysis (Chiral column: Column: Chiralpak OD-3, 3 µm, 0.46 cm id × 5 cm L; Mobile phase: A: CO ₂ , B: MeOH (0.05% IPAm); Gradient: B/ A = 10% - 40% for 3 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm), ee% was 100%.

Example 3. WX003

Synthesis path:

Step 1: Synthesis of Compound WX003

2,4,6-tributyl-1,3, 5,2,4,6-trioxatrifosfinan-2,4,6-trioxide (456.13 mg, 716.78 µmol, 426.29 µl, 50% purity in ethyl acetate, 2.5 eq) and N,N-diisopropylethylamine (111.16 mg, 860.13 µmol, 149.82 μl, 3 eq.), and the mixture was stirred at 25°C for 10 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. 30 ml of ethyl acetate was added to the reaction solution, and the mixture was washed with 20 ml of a saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Xtimate C18 150 mm*25mm*5 µm; Mobile phase: [water (10 mM NH ₄ HCO ₃ )-ACN]; acetonitrile%: 35%-65%, 10.5 min) to give connection WX003. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.63 (broad s, 1 H), 8.46 (dd, J = 4.63, 1.54 Hz, 1 H), 7.79 (d, J = 7.94 Hz, 1 H ), 7.48 (dd, J = 8.05, 4.74 Hz, 1 H), 7.23 - 7.37 (m, 5 H), 5.01 (dt, J = 10.80, 7.28 Hz, 1 H), 4.43 - 4.72 (m, 2 H ), 4.14 (broad s, 2 H), 3.80 (s, 3 H); LCMS m/z = 395.2 [M+H] ⁺ . According to SLC analysis (Chiral column: Column: Chiralpak OD -3, 3 µm, 0.46 cm id × 5 cm L; Mobile phase: A: CO ₂ , B: MeOH (0.05% IPAm); Gradient: B/A = 10% - 40% for 3 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm), ee% was 96.3%.

Example 4. WX004

Synthesis path:

Step 1: Synthesis of Compound WX004

2,4,6-tributyl-1,3, 5,2,4,6-trioxatrifosfinan-2,4,6-trioxide (456.13 mg, 716.78 µmol, 426.29 µl, 50% purity in ethyl acetate, 2.5 eq.) and N,N-diisopropylethylamine (111.16 mg, 860.14 µmol, 149.82 μl, 3 eq.), and the mixture was stirred at 25°C for 10 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. 30 ml of ethyl acetate was added to the reaction solution, and the mixture was washed with 20 ml of a saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Xtimate C18 15 mm*25 mm*5 µm; Mobile phase: [water (10 mM NH ₄ HCO ₃ )-ACN]; acetonitrile%: 35%-65%, 10.5 min), receiving connection WX004. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.73 (broad s, 1 H), 8.46 (d, J = 4.85 Hz, 1 H), 7.69 - 7.86 (m, 3 H), 7.44 - 7.65 (m, 3 H), 5.01 (dt, J = 11.08, 7.14 Hz, 1 H), 4.48 - 4.68 (m, 2 H), 4.22 (s, 2 H), 3.80 (s, 3 H); LCMS m/z = 420.1 [M+H] ⁺ . According to SLC analysis (Chiral column: Column: Chiralpak OD -3, 3 µm, 0.46 cm id × 5 cm L; Mobile phase: A: CO ₂ , B: MeOH (0.05% IPAm); Gradient: B/A = 10% - 40% for 3 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm), ee% was 100%.

Example 5. WX005

Synthesis path:

Step 1: Synthesis of Compound WX005

2,4,6-tributyl-1,3, 5,2,4,6-trioxatrifosfinan-2,4,6-trioxide (599.53 mg, 942.12 µmol, 560.31 µl, 50% purity in ethyl acetate, 2.5 eq.) and N,N-diisopropylethylamine (182.64 mg, 1.41 mmol, 246.15 μl, 3 eq.), and the mixture was stirred at 25°C for 10 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. Ethyl acetate (30 ml) was added to the reaction solution, and the mixture was washed with a saturated aqueous sodium chloride solution (20 ml). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Phenomenex Luna C18 150 mm*30 mm*5 µm; Mobile phase: [water (0.05% HCl)-ACN]; acetonitrile%: 20%-50%, 12 min) to give the compound WX005. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.62 (broad s, 1H), 8.46 (dd, J = 1.75, 4.82 Hz, 1H), 7.79 (dd, J = 1.32, 7.89 Hz, 1H) , 7.48 (dd, J = 4.82, 7.89 Hz, 1H), 7.20–7.37 (m, 5H), 5.01 (td, J = 7.34, 11.18 Hz, 1H), 4.60 (broad s, 1H), 4.49–4.55 (m, 1H), 4.14 (broad s, 2H); LCMS m/z = 398.2 [M+H] ⁺ . According to SLC analysis (Chiral column: Column: Chiralpak OD -3, 3 µm, 0.46 cm id × 5 cm L; Mobile phase: A: CO ₂ , B: MeOH (0.05% IPAm); Gradient: B/A = 10% - 40% for 3 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm), ee% was 100%.

Example 6. WX006

Synthesis path:

Step 1: Synthesis of Compound WX006

2,4,6-tributyl-1,3, 5,2,4,6-trioxatrifosfinan-2,4,6-trioxide (60.82 mg, 95.57 µmol, 56.84 µl, 50% purity in ethyl acetate, 2.5 eq.) and N,N-diisopropylethylamine (18.53 mg, 143.36 µmol, 24.97 μl, 3 eq.), and the mixture was stirred at 25°C for 10 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was directly purified by preparative HPLC (Column: Phenomenex Luna C18 150 mm*30 mm*5 µm; Mobile phase: [water (0.05% HCl)-ACN]; acetonitrile%: 20%-50%, 12 min) to obtain connection WX006. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 9.01 (br. s, 1H), 8.34-8.46 (m, 2H), 7.51-7.64 (m, 2H), 7.43 (br. s, 3H), 7.28- 7.31 (m, 2H), 7.22-7.26 (m, 1H), 5.29 (br.s, 2H), 5.20 (b.d, J = 7.02 Hz, 1H), 4.58-4.69 (m, 2H), 3.91 ( s, 3H); LCMS m/z = 394.2 [M+H] ⁺ . According to SLC analysis (Chiral column: Column: Chiralpak OD -3, 3 µm, 0.46 cm id × 5 cm L; Mobile phase: A: CO ₂ , B: MeOH (0.05% IPAm); Gradient: B/A = 10% - 40% for 3 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm), ee% was 100%.

Example 7. WX007

Synthesis path:

Step 1: Synthesis of Compound WX007

2,4,6-tributyl-1,3, 5,2,4,6-trioxatrifosfinan-2,4,6-trioxide (60.82 mg, 95.57 µmol, 56.84 µl, 50% pure in ethyl acetate, 2.5 eq) and N,N-diisopropylethylamine (18.53 mg, 143.36 µmol, 24.97 μl, 3 eq.), and the mixture was stirred at 25°C for 10 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was directly purified by preparative HPLC (Column: Phenomenex Luna C18 150 mm*30 mm*5 µm; Mobile phase: [water (0.05% HCl)-ACN]; acetonitrile%: 20%-50%, 12 min) to obtain connection WX007. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.98 (d, J = 8.33 Hz, 1H), 8.46 (dd, J = 1.32, 4.82 Hz, 1H), 7.77 (dd, J = 1.75, 7.89 Hz , 1H), 7.47 (dd, J = 4.60, 8.11 Hz, 1H), 7.22–7.40 (m, 5H), 6.54 (s, 1H), 5.00 (td, J = 7.45, 11.40 Hz, 1H), 4.59– 4.70 (m, 1H), 4.48-4.56 (m, 1H), 4.22 (s, 2H), 3.79 (s, 3H); LCMS m/z = 395.1 [M+H] ⁺ . According to SLC analysis (Chiral column: Column: Chiralpak OD -3, 3 µm, 0.46 cm id × 5 cm L; Mobile phase: A: CO ₂ , B: MeOH (0.05% IPAm); Gradient: B/A = 10% - 40% for 3 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm), ee% was 100%.

Example 8. WX008

Synthesis path:

Step 1: Synthesis of Compound WX008

2,4,6-tributyl-1,3, 5,2,4,6-trioxatrifosfinan-2,4,6-trioxide (60.82 mg, 95.57 µmol, 56.84 µl, 50% pure in ethyl acetate, 2.5 eq) and N,N-diisopropylethylamine (18.53 mg, 143.36 µmol, 24.97 μl, 3 eq.), and the mixture was stirred at 25°C for 10 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was directly purified by preparative HPLC (Column: Phenomenex Luna C18 150 mm*30 mm*5 µm; Mobile phase: [water (0.05% HCl)-ACN]; acetonitrile%: 20%-50%, 12 min) to obtain connection WX008. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.85 (d, J = 7.89 Hz, 1H), 8.46 (d, J = 3.07 Hz, 1H), 7.75-7.89 (m, 3H), 7.61 (d , J = 4.82 Hz, 2H), 7.48 (dd, J = 4.82, 7.89 Hz, 1H), 5.58 (s, 2H), 4.94–5.06 (m, 1H), 4.65 (t, J = 10.74 Hz, 1H) , 4.45-4.57 (m, 1H), 3.80 (s, 3H); LCMS m/z = 454.1 [M+H] ⁺ . According to SLC analysis (Chiral column: Column: Chiralpak OD -3, 3 µm, 0.46 cm id × 5 cm L; Mobile phase: A: CO ₂ , B: MeOH (0.05% IPAm); Gradient: B/A = 10% - 40% for 3 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm), ee% was 100%.

Example 9. WX009

Synthesis path:

Step 1: Synthesis of Compound WX009

2,4,6-tributyl-1,3, 5,2,4,6-trioxatrifosfinan-2,4,6-trioxide (60.82 mg, 95.57 µmol, 56.84 µl, 50% pure in ethyl acetate, 2.5 eq) and N,N-diisopropylethylamine (18.53 mg, 143.36 µmol, 24.97 μl, 3 eq.), and the mixture was stirred at 25°C for 10 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was directly purified by preparative HPLC (Column: Phenomenex Luna C18 150 mm*30 mm*5 µm; Mobile phase: [water (0.05% HCl)-ACN]; acetonitrile%: 20%-50%, 12 min) to obtain connection WX009. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.84 (s, 1H), 8.71 (d, J = 7.89 Hz, 1H), 8.46 (dd, J = 1.53, 4.60 Hz, 1H), 7.79 (dd , J = 1.53, 8.11 Hz, 1H), 7.48 (dd, J = 4.82, 7.89 Hz, 1H), 7.26–7.42 (m, 5H), 5.49 (s, 2H), 4.94–5.08 (m, 1H), 4.48-4.63 (m, 2H), 3.80 (s, 3H); LCMS m/z = 395.1 [M+H] ⁺ . According to SLC analysis (Chiral column: Column: Chiralpak OD -3, 3 µm, 0.46 cm id × 5 cm L; Mobile phase: A: CO ₂ , B: MeOH (0.05% IPAm); Gradient: B/A = 10% - 40% for 3 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm), ee% was 100%.

Example 10. WX010

Synthesis path:

Step 1: Synthesis of Compound WX010

2,4,6-tributyl-1,3, 5,2,4,6-trioxatrifosfinan-2,4,6-trioxide (60.82 mg, 95.57 µmol, 56.84 µl, 50% purity in ethyl acetate, 2.5 eq) and N,N-diisopropylethylamine (18.53 mg, 143.36 µmol, 24.97 μl, 3 eq.), and the mixture was stirred at 25°C for 10 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was directly purified by preparative HPLC (Column: Phenomenex Luna C18 150 mm*30 mm*5 µm; Mobile phase: [water (0.05% HCl)-ACN]; acetonitrile%: 20%-50%, 12 min) to obtain connection WX010. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.75 (d, J = 8.33 Hz, 1H), 8.69 (s, 1H), 8.46 (dd, J = 1.53, 4.60 Hz, 1H), 7.79 (dd , J = 1.53, 8.11 Hz, 1H), 7.48 (dd, J = 4.82, 7.89 Hz, 1H), 7.30–7.41 (m, 5H), 5.65 (s, 2H), 4.97–5.10 (m, 1H), 4.58-4.68 (m, 1H), 4.47-4.57 (m, 1H), 3.80 (s, 3H); LCMS m/z = 395.1 [M+H] ⁺ . According to SLC analysis (Chiral column: Column: Chiralpak OD -3, 3 µm, 0.46 cm id × 5 cm L; Mobile phase: A: CO ₂ , B: MeOH (0.05% IPAm); Gradient: B/A = 10% - 40% for 3 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm), ee% was 100%.

Example 11. WX011

Synthesis path:

Step 1: Synthesis of Compound WX011

2,4,6-tributyl-1,3, 5,2,4,6-trioxatrifosfinan-2,4,6-trioxide (60.82 mg, 95.57 µmol, 56.84 µl, 50% purity in ethyl acetate, 2.5 eq.) and N,N-diisopropylethylamine (18.53 mg, 143.36 µmol, 24.97 μl, 3 eq.), and the mixture was stirred at 25°C for 10 hours. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was directly purified by preparative HPLC (Column: Phenomenex Luna C18 150 mm*30 mm*5 µm; Mobile phase: [water (0.05% HCl)-ACN]; acetonitrile%: 20%-50%, 12 min) to obtain connection WX011. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.83 (d, J = 7.89 Hz, 1H), 8.41–8.49 (m, 1H), 7.77 (dd, J = 1.32, 7.89 Hz, 1H), 7.46 (dd, J = 4.82, 7.89 Hz, 1H), 7.30-7.42 (m, 3H), 7.26 (d, J = 7.02 Hz, 2H), 5.48 (s, 2H), 4.98 (td, J = 7.13, 11.62 Hz, 1H), 4.59–4.67 (m, 1H), 4.49 (dd, J = 6.80, 9.87 Hz, 1H), 3.78 (s, 3H); LCMS m/z = 429.0 [M+H] ⁺ . According to SLC analysis (Chiral column: Column: Chiralpak OD -3, 3 µm, 0.46 cm id × 5 cm L; Mobile phase: A: CO ₂ , B: MeOH (0.05% IPAm); Gradient: B/A = 10% - 40% for 3 minutes; Flow rate: 4.0 ml/min; Wavelength: 220 nm), ee% was 100%.

Example 12. WX012

Synthesis path:

Step 1: Synthesis of Compound WX012

Trimethylaluminum (2M, 53.76 µl, 1.5 eq.) and the mixture was stirred at 25°C for 1 hour. According to LCMS monitoring, the original compounds completely reacted, and the signal of the target product was observed. The reaction solution was directly purified by preparative HPLC (Column: Phenomenex Luna C18 150 mm*30 mm*5 µm; Mobile phase: [water (0.05% HCl)-ACN]; acetonitrile%: 20%-50%, 12 min) to obtain connection WX012. LCMS m/z = 396.2 [M+H] ⁺ .

Example 13. WX013

Synthesis path:

Step 1: Synthesis of Compound WX013-1

LiOH⋅H ₂ O (245.59 mg, 5.85 mmol, 3 equiv.) was added to a solution of BB-11B (501.93 mg, 1.95 mmol, 1 equiv.) in tetrahydrofuran (9 ml) and water (3 ml), and the mixture was heated at 25°C for 2 hours. Water (10 ml) and ethyl acetate (10 ml) were added to the reaction solution, and the layers were separated. The aqueous phase was extracted with ethyl acetate (10 ml*2). The resulting aqueous phase was adjusted to pH 5-6 with 1M HCl solution and extracted with ethyl acetate (10 ml*2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure (water jet pump, 45°C) to give WX013-1. LCMS m/z = 230.2 [M+1] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 12.75 - 13.57 (m, 1 H), 7.33 - 7.44 (m, 3 H), 7.18 - 7.28 (m, 2 H), 5.57 (dd, J = 7.97, 6.21 Hz, 1 H), 3.06 - 3.22 (m, 2 H), 2.94 - 3.03 (m, 1 H), 2.53 - 2.61 (m, 1 H).

Step 2: Synthesis of Compound WX013

N,N-diisopropylethylamine (23.16 mg, 179.20 µmol, 31.21 µl, 2.5 eq.) and WX013-1 (24.65 mg, 107.52 µmol, 1.5 eq.) were added to a solution of BB-1 (15 mg, 71.68 µmol, 1 eq. ) in DMF (1.5 ml) and then propylphosphonic anhydride (T ₃ P, 68.42 mg, 107.52 µmol, 63.94 µl, 50% purity, 1.5 eq.) was added. The mixture was heated at 25°C for 1 hour. The reaction solution was filtered, and the filtrate was collected. The filtrate was purified by preparative HPLC (Column: Phenomenex Luna C18 150 mm*30 mm*5 µm; Mobile phase: [water (0.05% HCl)-ACN]; acetonitrile%: 20%-50%, 12 min) to give WX013. LCMS m/z = 421.1 [M+1] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.68 (d, J = 8.04 Hz, 1 H), 8.46 (dd, J = 4.72, 1.32 Hz, 1 H), 7.78 (dd, J = 8.04, 1.38 Hz, 1 H), 7.47 (dd, J = 8.04, 4.64 Hz, 1 H), 7.32 - 7.43 (m, 3 H), 7.24 (broad d, J = 6.65 Hz, 2 H), 5.44 - 5.68 (m, 1 H), 5.00 (dt, J = 11.40, 7.40 Hz, 1 H), 4.42 - 4.68 (m, 2 H), 3.79 (s, 3 H), 2.90 - 3.23 (m, 4 H).

Example 14. WX014

Synthesis path:

Step 1: Synthesis of Compound WX014-2

WX014-1 was dissolved in THF (100 ml) and MeOH (100 ml) and the reaction flask was placed in an ice bath (0°C). Trimethylsilyl diazomethane (2M, 47.89 ml, 2.1 eq.) was added to the flask, the mixture was heated to 25°C and stirred for 2 hours. Glacial acetic acid was added dropwise until the solution became pale yellow. The reaction solution was evaporated under reduced pressure to give compound WX014-2.

Step 2: Synthesis of Compound WX014-3

Triphenylphosphine (14.30 g, 54.53 mmol, 1.2 eq.) and imidazole (3.71 g, 54.53 mmol, 1.2 eq.) were dissolved in dichloromethane (120 ml) at 25°C and I ₂ (13.84 g, 54.53 mmol, 10.98 ml) was added slowly , 1.2 equiv.) into the resulting mixture. The mixture was stirred for 10 minutes and a solution of WX014-2 in DCM (80 ml) was added thereto. The mixture was heated at 25°C for 12 hours. The reaction solution was filtered, and the filtrate was evaporated under reduced pressure to give a crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate=10:1~1:1) to give WX014-3. LCMS m/z = 244.1 [M-100+1] ⁺ . ^1H NMR (400 MHz, _CDCl3 ) δ = 5.10 (broad.d, J = 4.82 Hz, 1H), 4.35 (broad.d, J = 3.96 Hz, 1H), 3.77 (s, 3H), 3.18 (t, J = 7.46 Hz, 2 H), 2.30 - 2.49 (m, 1 H), 2.18 (d.sq., J = 14.32, 7.22 Hz, 1 H), 1.43 - 1.47 (m, 9 H).

Step 3: Synthesis of Compound WX014-4

1,2-Dibromoethane (82.12 mg, 437.12 µmol, 32.98 µl, 0.3 equiv.) was added to zinc powder (285.83 mg, 4.37 mmol, 3 equiv.) in DMF (2 ml). The mixture was stirred at 60°C for 30 minutes and then cooled to room temperature (25°C). Trimethylchlorosilane (9.50 mg, 87.42 µmol, 11.10 µl, 0.06 eq.) was added to the reaction flask and the mixture was stirred at 25°C for 30 minutes. A solution of WX014-2 in DMF (2 ml) was added to the resulting mixture, and the mixture was heated at 25°C for 30 minutes, then left for 30 minutes. The upper emulsion layer was separated and slowly added to a solution of tris(dibenzylideneacetone)dipalladium (44.08 mg, 48.14 µmol, 0.05 equiv.), tri-o-methylphenylphosphine (58.61 mg, 192.55 µmol, 0.2 equiv.) and BB-12 in DMF (2 ml), and the mixture was heated at 20°C for 1 hour. Water (5 ml) and methyl tert-butyl ether (5 ml) were added to the reaction solution, and the layers were separated. The aqueous phase was extracted with methyl tert-butyl ether (5 ml*2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give WX014-4. LCMS m/z = 289.1 [M-56+1] ⁺ .

Step 4: Synthesis of Compound WX014-5

LiOH⋅H ₂ O (219.33 mg, 5.23 mmol, 3 eq.) was added to a solution of WX014-4 in tetrahydrofuran (15 ml) and water (15 ml) and the mixture was heated at 25°C for 2 hours. 10 ml of water and 10 ml of ethyl acetate were added to the reaction solution, and the layers were separated. The aqueous phase was extracted with ethyl acetate (10 ml*2). The resulting aqueous phase was adjusted to pH 5~6 with 1M HCl solution and extracted with ethyl acetate (10 ml*2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give WX014-5. LCMS m/z = 275.1 [M-56+1] ⁺ .

Step 5: Synthesis of Compound WX014-6

Propylphosphonic anhydride (895.80 mg, 1.41 mmol, 837.20 μl, 50% purity, 1.5 eq.) was added to a solution of N,N-diisopropylethylamine (303.22 mg, 2.35 mmol, 408.65 μl, 2.5 eq.) and WX014-5 in DMF (20 ml), and the mixture was heated at 25°C for 2 hours. Water (15 ml) and ethyl acetate (15 ml) were added to the reaction solution, and the layers were separated. The aqueous phase was extracted with ethyl acetate (15 ml*2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate=10:1~1:1) to obtain WX014-6. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.40 (br. s, 1 H), 6.76 - 6.87 (m, 2 H), 5.46 (br. d, J = 7.46 Hz, 1 H), 4.22 - 4.37 (m, 2 H), 2.88 - 3.03 (m, 1 H), 2.63 - 2.73 (m, 2 H), 1.42 (s, 9 H).

Step 6: Synthesis of Compound WX014-7

Lawsson's reagent (77.70 mg, 192.12 µmol, 1.5 equiv.) was added to a solution of WX014-6 (40 mg, 128.08 µmol, 1 equiv.) in toluene (5 ml) at 20°C, and the mixture was heated at 70°C for 12 hours. The reaction solution was evaporated under reduced pressure to give a crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate=10:1~1:1) to obtain WX014-7. LCMS m/z = 273.1 [M-56+1] ⁺ .

Step 7: Synthesis of Compound WX014-8

HCl/EtOAc (4M, 2 ml) was added to a solution of WX014-7 in ethyl acetate (3 ml) and the mixture was heated at 25°C for 2 hours. The reaction solution was evaporated under reduced pressure (water jet pump, 40° C.) to give crude product WX014-8. LCMS m/z = 229.2 [M+1] ⁺ .

Step 8: Synthesis of Compound WX014

DIPEA (18.31 mg, 141.66 µmol, 24.67 µl, 2.5 equiv.) and T ₃ P (54.09 mg, 84.99 µmol, 50.55 µl, 50% purity, 1.5 equiv.) were added to WX014-8 solution (15 mg, 56.66 µmol, 1 eq., HCl) and BB-1 (14.97 mg, 73.66 µmol, 1.3 eq.) in DMF (2 ml) and the mixture was heated at 25°C for 2 hours. Water (5 ml) and ethyl acetate (5 ml) were added to the reaction solution, and the layers were separated. The aqueous phase was extracted with ethyl acetate (5 ml*2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Phenomenex Luna C18 150mm*30mm*5µm; Mobile phase: [water (0.05% HCl)-ACN]; acetonitrile%: 20%-50%, 12 min) to give WX014. LCMS m/z = 414.2 [M+1] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ = 7.21 - 7.37 (m, 5 H), 6.97 - 7.13 (m, 2 H), 4.71 (brd, J = 4.14 Hz, 1 H), 4.18 ( brs, 2 H), 2.84–2.96 (m, 1 H), 2.64–2.83 (m, 2 H), 2.17 (td, J = 11.67, 7.78 Hz, 1 H).

Example 15. WX015

Synthesis path:

Step 1: Synthesis of Compound WX015

BB-3 (20 mg, 96.02 µmol, 1.1 eq.) and DIPEA (23.69 mg, 183.32 µmol, 31.93 µl, 2.1 eq.) were added to a round bottom flask containing N,N-dimethylformamide (10 ml). After dissolution, BB-14 (21.24 mg, 87.30 µmol, 1 eq.) and T ₃ P (55.55 mg, 87.30 µmol, 51.92 µl, 50% pure, 1 eq.) were added, and then the mixture was left to run the reaction with stirring under 25°C for 12 hours. LCMS analysis showed the almost complete disappearance of the original substances. The reaction solution was evaporated under reduced pressure to a volume of about 5 ml. HPLC (Column: Welch Xtimate C18 150 mm*25 mm*5 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 35%-65%, 8 min) was used for purification to give product WX015 . ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.78 (broad.d, J = 7.5 Hz, 1H), 7.43 - 7.35 (m, 5H), 7.35 - 7.26 (m, 4H), 5.82 (s, 2H) , 5.21 (td, J = 6.9, 11.0 Hz, 1H), 4.71 (dd, J = 6.5, 9.5 Hz, 1H), 4.33–4.21 (m, 1H), 3.87 (s, 3H); LCMS m/z = 395.1 [M+1] ⁺ .

Example 16. WX016

Synthesis path:

Step 1: Synthesis of Compound WX016-2

DMSO (10 ml) was placed in a round bottom flask and then WX016-1 (1.31 g, 8.44 mmol, 1 eq.), K ₂ CO ₃ (1.40 g, 10.13 mmol, 1.2 eq.) and benzyl bromide (1.59 g, 9.29 mmol, 1.10 ml, 1.1 equiv.). The mixture was continuously stirred at 25°C for 12 hours. After completion of the reaction, water (50 ml) was added to the reaction solution, and the mixture was extracted twice with ethyl acetate (30 ml×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (5 ml × 2), and evaporated under reduced pressure to give the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate=100:1~1:1) to give WX016-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.39 - 7.30 (m, 3H), 7.19 (dd, J = 1.8, 7.6 Hz, 2H), 5.39 (s, 2H), 4.49 (q, J = 7.1 Hz , 2H), 2.43 (s, 3H), 1.44 (t, J = 7.1 Hz, 3H); LCMS m/z = 246.1 [M+1] ⁺ .

Step 2: Synthesis of Compound WX016-3

THF (7.5 ml) and H ₂ O (2.5 ml) were placed in a round bottom flask and then WX016-2 (1 g, 4.08 mmol, 1 equiv.) and LiOH⋅H ₂ O (513.26 mg, 12.23 mmol, 3 equiv. .). The mixture was left to pass the reaction with stirring at 25°C for 2 hours. The reaction solution was evaporated under reduced pressure, and 2N HCl was added dropwise. hydrochloric acid solution, adjusting the pH to acidic. The mixture was filtered to give a filter cake which was the title compound WX016-3. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.42 - 7.28 (m, 3H), 7.25 - 7.19 (m, 2H), 5.40 (s, 2H), 2.56 (s, 3H); LCMS m/z = 217.9 [M+1] ⁺ .

Step 3: Synthesis of Compound WX016

N,N-dimethylformamide (1 mL), WX016-3 (80 mg, 368.28 µmol, 1 eq.) and BB-4 (77.07 mg, 368.28 µmol, 1 eq.) were placed in a round-bottom flask, then T ₃ P was added ( 351.54 mg, 552.43 µmol, 328.55 µl, 50% purity, 1.5 eq.) and DIPEA (142.79 mg, 1.10 mmol, 192.45 µl, 3 eq.). The mixture was continuously stirred at 25°C for 5 hours, then evaporated to give the crude product. The crude product was purified by preparative HPLC (Column: Welch Xtimate C18 150 mm*25 mm*5 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 35%-65%, 8 min) to give connection WX016. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.72 (broad d, J = 7.3 Hz, 1H), 8.37 (dd, J = 1.4, 4.6 Hz, 1H), 7.56 (dd, J = 1.3, 8.0 Hz , 1H), 7.38 - 7.27 (m, 4H), 7.18 (bd, J = 6.0 Hz, 2H), 5.36 (s, 2H), 5.23 (td, J = 6.8, 11.0 Hz, 1H), dd, J = 6.5, 9.5 Hz, 1H), 4.32 (dd, J = 9.8, 11.0 Hz, 1H), 3.93 (s, 3H), 2.41 (s, 3H); LCMS m/z = 409.1 [M+1] ⁺ .

Example 17. WX017

Synthesis path:

Step 1: Synthesis of Compound WX017-2

Lithium aluminum hydride (629.99 mg, 16.60 mmol, 2 equiv.) and THF (5 ml) were placed into the reaction flask at 25°C, then a solution of WX017-1 (1.08 g, 8.30 mmol, 1 equiv.) in THF was slowly added dropwise (5 ml). After the addition was completed, the mixture was heated to 35° C. and stirred continuously for 1 hour. The reaction was quenched by adding 6 ml of a saturated aqueous solution of Na ₂ SO ₄ . The mixture was filtered and washed with ethyl acetate (10 ml x 3). The organic phases were combined, dried over anhydrous MgSO ₄ and filtered. The filtrate was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~5:1) to give compound WX017-2. ^1H NMR (400 MHz, _CDCl3 ) δ = 3.84 (s, 2H), 2.01 (d, J = 2.5 Hz, 6H).

Step 2: Synthesis of Compound WX017-3

WX017-2, DCM (10 ml) and pyridine (1.40 g, 17.70 mmol, 1.43 ml, 4.11 equiv.) were placed in a reaction flask, then p-toluenesulfonyl chloride (1.23 g, 6.46 mmol, 1.5 equiv.) was slowly added. The mixture was heated to 40°C and left to pass the reaction for 24 hours. A new spot (Rf = 0.7) appeared on TLC (petroleum ether:ethyl acetate = 5:1). The reaction solution was immediately evaporated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~30:1) to obtain compound WX017-3. ^1H NMR (400 MHz, _CDCl3 ) δ = 7.78 (d, J = 8.3 Hz, 2H), 7.35 (d, J = 8.0 Hz, 2H), 4.24 (s, 2H), 2.46 (s, 3H), 1.99 (d, J = 2.3 Hz, 6H).

Step 3: Synthesis of Compound WX017-4

Acetonitrile (10 mL), WX017-3 (500 mg, 1.85 mmol, 1.1 equiv.) and BB-8-1 (213.72 mg, 1.68 mmol, 1 equiv.) were placed into a pear-shaped flask at 25°C, then K ₂ was added CO ₃ (278.88 mg, 2.02 mmol, 1.2 equiv.). The mixture was heated to 70° C. and left to react for 48 hours. The reaction solution was evaporated, and the crude product was purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate=100:1~1:2) to obtain compound WX017-4. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.12 (s, 1H), 4.57 (s, 2H), 4.01 (s, 3H), 2.05 (d, J = 2.5 Hz, 6H).

Step 4: Synthesis of Compound WX017-5

WX017-4 (100 mg, 444.01 µmol, 1 eq.) was added to a round bottom flask containing THF (2 ml) at 25°C. After dissolution, a solution of LiOH⋅H ₂ O (55.90 mg, 1.33 mmol, 3 eq.) in MeOH (1 ml) was added dropwise and the mixture was heated at 25°C for 5 hours. The mixture was evaporated under reduced pressure and the residue was dissolved in tetrahydrofuran (5 ml). Added 1n. hydrochloric acid solution, adjusting the pH to 4, then toluene (5 ml) was added. The mixture was stirred well and the solvent was evaporated to dryness under reduced pressure to give crude product WX017-5, which was directly used in the next step without purification.

Step 5: Synthesis of Compound WX017

Ethyl acetate (10 mL), WX017-5 (207 mg, 588.09 µmol, 1 eq.) and BB-4 (143.52 mg, 588.09 µmol, 1 eq.) were placed in a reaction flask at 25°C. After dissolution, T ₃ P (1.12 g, 1.76 mmol, 1.05 ml, 50% purity, 3 eq.) and DIPEA (228.02 mg, 1.76 mmol, 307.30 μl, 3 eq.) were added and the mixture was heated at 25°C for 8 ocloc'k. After completion of the reaction, ethyl acetate (10 ml) was added. The organic phase was washed with water (10 ml×3), then with saturated aqueous sodium chloride solution (10 ml×1) and evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Welch Xtimate C18 150 mm*25 mm*5 µm; Mobile phase: [water (0.05% NH ₃ H ₂ O)-ACN]; acetonitrile%: 35%-65%, 8 min ) to get a WX017 connection. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.83 (broad.d, J = 6.8 Hz, 1H), 8.37 (dd, J = 1.4, 4.6 Hz, 1H), 8.08 (s, 1H), 7.57 (dd , J = 1.3, 8.0 Hz, 1H), 7.29 (dd, J = 4.8, 8.0 Hz, 1H), 5.20 (td, J = 6.7, 11.2 Hz, 1H), 4.80 (dd, J = 6.4, 9.4 Hz, 1H), 4.54 (s, 2H), 4.33 (dd, J = 9.8, 11.0 Hz, 1H), 3.94 (s, 3H), 2.05 (d, J = 2.3 Hz, 6H); LCMS m/z = 403.2 [M+1] ⁺ .

Example 18. WX018

Synthesis path:

Synthesis of compound WX018

BB-4 (200 mg, 955.71 µmol, 1.1 eq.) and DIPEA (235.80 mg, 1.82 mmol, 317.80 µl, 2.1 eq.) were added to a round bottom flask containing N,N-dimethylformamide (10 ml). After dissolution, BB-14 (210.50 mg, 868.83 µmol, 1 eq) and T ₃ P (552.89 mg, 868.83 µmol, 516.72 µl, 50% pure, 1 eq) were added and the mixture was stirred at 25°C for 12 hours. The reaction solution was evaporated under reduced pressure to a volume of about 5 ml. HPLC (Column: Welch Xtimate C18 150 mm*25 mm*5 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 35%-65%, 8 min) was used for purification to give compound WX018 . ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.86 (broad.d, J = 6.8 Hz, 1H), 8.38 (dd, J = 1.6, 4.6 Hz, 1H), 7.69 - 7.27 (m, 7H), 5.83 (s, 2H), 5.20 (td, J = 6.7, 11.2 Hz, 1H), 4.78 (dd, J = 6.3, 9.5 Hz, 1H), 4.33 (dd, J = 9.7, 11.2 Hz, 1H), 3.93 ( s, 3H); LCMS m/z = 396.1 [M+1] ⁺ .

Example 19. WX019

Synthesis path:

Step 1: Synthesis of Compound WX019

BB-15 (156.11 mg, 745.97 µmol, 1.1 equiv.), BB-4 (150 mg, 678.16 µmol, 1 equiv.), T ₃ P ( 431.55 mg, 678.16 µmol, 403.32 µl, 50% pure, 1 eq.) and DIPEA (184.06 mg, 1.42 mmol, 248.06 µl, 2.1 eq.), then stirring was started. The mixture was allowed to react with stirring at 25° C. for 12 hours. After completion of the reaction, the solution was evaporated under reduced pressure to a volume of about 5 ml. HPLC (Column: Welch Xtimate C18 150 mm*25 mm*5 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 35%-65%, 8 min) was used for purification to give product WX019 . ^1H NMR (400 MHz, _CDCl3 ) δ = 8.81 (broad.d, J = 7.3 Hz, 1H), 8.37(dd, J = 1.5, 4.8 Hz, 1H), 8.14 (s, 1H), 7.56 (dd , J = 1.5, 8.0 Hz, 1H), 7.41 - 7.27 (m, 3H), 7.21 - 7.06 (m, 2H), 5.45 (s, 2H), 5.20 (td, J = 6.7, 11.2 Hz, 1H), 4.78 (dd, J = 6.5, 9.5 Hz, 1H), 4.31 (dd, J = 9.5, 11.0 Hz, 1H), 3.93 (s, 3H); ¹⁹ F NMR (376 MHz, CDCl3) δ = -109.59 ~ -130.05 (m, 1F); LCMS m/z = 413.1 [M+1] ⁺ .

Example 20. WX020

Synthesis path:

Step 1: Synthesis of Compound WX020-2

WX020-1 (3 g, 23.60 mmol, 1 equiv.), 2,6-difluorobenzyl bromide (5.37 g, 25.96 mmol, 1.1 equiv.) and K ₂ CO ₃ (3.91 g, 28.32 mmol, 1.2 eq.), then stirring was started. The mixture was heated at 50°C for 12 hours. After the end of the reaction according to TLC monitoring, the reaction solution was filtered to remove K ₂ CO ₃ and the filter cake was washed three times with ethyl acetate (15 ml). Saturated aqueous sodium chloride solution (50 ml) was added to the filtrate and the organic phase was separated after extraction. The aqueous phase was extracted with ethyl acetate (30 ml) again. The organic phases were combined and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate=100:1~100:100) to obtain compound WX020-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.17 (s, 1H), 7.46 - 7.33 (m, 1H), 7.04 - 6.91 (m, 2H), 5.51 (s, 2H), 4.13 - 3.85 (m, 3H).

Stage 2: synthesis of compound WX020-3

WX020-2 (1 g, 3.95 mmol, 1 equiv.) was added to a round bottom flask containing THF (30 mL). After dissolution, a solution of LiOH⋅H ₂ O (331.43 mg, 7.90 mmol, 2 eq.) in H ₂ O (5 ml) was added and the mixture was left to react with stirring at 20°C for 2 hours. The reaction solution was adjusted to pH 6 by adding 1M hydrochloric acid, and the mixture was evaporated under reduced pressure to remove the solvent to give crude product WX020-3; LCMS m/z = 239.9 [M+1] ⁺ .

Step 3: Synthesis of Compound WX020

BB-4 (192.49 mg, 919.82 µmol, 1.1 equiv.), WX020-3 (200 mg, 836.20 µmol, 1 equiv.), T ₃ P( 532.12 mg, 836.20 µmol, 497.31 µl, 50% pure, 1 eq.) and DIPEA (226.95 mg, 1.76 mmol, 305.86 µl, 2.1 eq.), and the mixture was left to run the reaction with stirring at 25°C for 12 hours . The reaction solution was evaporated under reduced pressure to a volume of about 5 ml, and the residue was purified by preparative HPLC (Column: Welch Xtimate C18 150 mm*25 mm*5 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile % : 35%-65%, 8 min) to give compound WX020. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.79 (broad.d, J = 7.0 Hz, 1H), 8.36 (dd, J = 1.5, 4.8 Hz, 1H), 8.14 (s, 1H), 7.55 (dd , J = 1.5, 8.0 Hz, 1H), 7.43 - 7.26 (m, 2H), 6.97 (t, J = 7.9 Hz, 2H), 5.49 (s, 2H), 5.19 (td, J = 6.7, 11.1 Hz, 1H), 4.77 (dd, J = 6.3, 9.5 Hz, 1H), 4.30 (dd, J = 9.7, 10.9 Hz, 1H), 3.92 (s, 3H); ¹⁹ F NMR (376 MHz, CDCl ₃ ) δ = -104.74 ~ -120.36 (m, 1F); LCMS m/z =431.1 [M+1] ⁺ .

Example 21. WX021

Synthesis path:

Step 1: Synthesis of Compound WX021-1

To a round bottom flask containing DCM (20 ml) was added BB-3-4 (2 g, 6.73 mmol, 1 eq.) and the mixture was stirred at 40°C. NCS (988.11 mg, 7.40 mmol, 1.1 eq.) was added slowly, then the reaction mixture was continuously stirred at 40°C for 5 hours. The reaction solution was evaporated under reduced pressure to give a crude product. The crude product was purified by silica gel column chromatography (eluent: DCM:MeOH=100:1~100:10) to give compound WX021-1. ¹ H NMR (400 MHz, CD ₃ OD) δ = 7.54 (d, J = 1.5 Hz, 1H), 7.34 (d, J = 1.8 Hz, 1H), 6.95 - 6.77 (m, 1H), 5.49 (s, 2H), 4.70 - 4.60 (m, 2H), 4.53 (broad dd, J = 4.3, 9.5 Hz, 1H), 4.39 - 4.26 (m, 1H), 1.46 (s, 9H). LCMS m/z = 331.9 [M+1] ⁺ .

Step 2: Synthesis of Compound WX021-2

To a round bottom flask containing DCM (10 ml) was added WX021-1 (1.60 g, 4.82 mmol, 1 eq.), T ₃ P (6.14 g, 9.65 mmol, 5.74 ml, 50% pure, 2 eq.) and DIPEA (1.87 g, 14.47 mmol, 2.52 ml, 3 eq.), and the mixture was heated at 20°C for 3 hours. The reaction solution was poured into a saturated sodium chloride aqueous solution (300 ml), and the layers were separated to separate the organic phase. The aqueous phase was extracted twice with ethyl acetate (100 ml). The organic phases were combined and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:20~100:100) to give compound WX021-2. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.22 (broad s, 1H), 8.04 (d, J = 2.0 Hz, 1H), 7.41 (d, J = 2.3 Hz, 1H), 5.59 (broad s , 1H), 4.71 - 4.46 (m, 2H), 4.23 - 4.05 (m, 1H), 1.46 (s, 9H); LCMS m/z = 257.8 [M-56+1] ⁺ .

Step 3: Synthesis of Compound WX021-3

To a round bottom flask containing N,N-dimethylacetamide (10 ml) was added WX021-2 (200 mg, 637.48 µmol, 1 eq.), iodomethane (1.17 g, 8.24 mmol, 513.16 µl, 12.93 eq.) and cesium carbonate ( 415.41 mg, 1.27 mmol, 2 equiv.). The mixture was allowed to react with stirring at 25° C. under nitrogen for 1 hour. After completion of the reaction, the solution was poured into a saturated aqueous sodium chloride solution (20 ml) and the mixture was extracted three times with ethyl acetate (15 ml). The organic phases were combined and evaporated under reduced pressure to give the crude product. The crude product was dissolved in ethyl acetate (2 ml) and purified by preparative TLC (petroleum ether:ethyl acetate=3:1) to give product WX021-3. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.23 (d, J = 2.3 Hz, 1H), 7.47 (d, J = 2.0 Hz, 1H), 5.55 (broad s, 1H), 4.82 - 4.49 (m , 2H), 4.35 - 4.14 (m, 1H), 3.47 (s, 3H), 1.41 (s, 9H).

Step 4: Synthesis of Compound WX021-4

WX021-3 (177 mg, 540.02 µmol, 1 equiv.) and Lawsson's reagent (327.63 mg, 810.04 µmol, 1.5 equiv.) were added to a round bottom flask containing toluene (10 ml) at 25°C, the mixture was slowly heated at 110 °C and left to pass the reaction for 12 hours. The reaction solution was poured into a saturated sodium chloride aqueous solution (100 ml), and the mixture was extracted three times with ethyl acetate (30 ml). The organic phases were combined and evaporated under reduced pressure to give the crude product. The crude product was purified by preparative TLC (petroleum ether:ethyl acetate=3:1) to give compound WX021-4. LCMS m/z = 287.8 [M-56+1] ⁺ .

Step 5: Synthesis of Compound WX021-5

To a round bottom flask containing dichloromethane (5 ml) was added WX021-4 (35 mg, 101.80 µmol, 1 eq.) at 25°C. Trifluoroacetic acid (174.10 mg, 1.53 mmol, 113.06 µl, 15 eq.) was added dropwise with stirring and the mixture was allowed to react for 2 hours. The reaction solution was evaporated to dryness under reduced pressure to give crude product WX021-5, which was directly used in the next step.

Step 6: Synthesis of Compound WX021

WX021-5 (20 mg, 82.06 µmol, 1 eq.), BB-8 (16.68 mg, 82.06 µmol, 1 eq.), T ₃ P ( 52.22 mg, 82.06 µmol, 48.81 µl, 50% pure, 1 eq.) and DIPEA (22.27 mg, 172.33 µmol, 30.02 µl, 2.1 eq.), and the mixture was heated at 25°C for 5 hours. Saturated aqueous sodium chloride solution (30 ml) was added and the mixture was extracted with ethyl acetate (30 ml×3). The organic phase was evaporated under reduced pressure to give the crude product. The crude product was purified by HPLC (formic acid system) (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase [water (0.225% formic acid)-ACN]; acetonitrile%: 40%-70%, 8 min ) receiving a WX021 connection. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.77 (broad.d, J = 7.3 Hz, 1H), 8.32 (d, J = 2.3 Hz, 1H), 8.04 (s, 1H), 7.57 (d, J = 2.0 Hz, 1H), 7.44 - 7.34 (m, 3H), 7.32 - 7.27 (m, 2H), 5.40 (s, 2H), 5.26 - 5.16 (m, 1H), 4.77 (dd, J = 6.3, 9.5 Hz, 1H), 4.39 - 4.27 (m, 1H), 3.90 (s, 3H); LCMS m/z = 429.0 [M+1] ⁺ .

Example 22. WX022

Synthesis path:

Step 1: Synthesis of Compound WX022

WX021-5 (20 mg, 82.06 µmol, 1 equiv.), BB-15 (18.15 mg, 82.06 µmol, 1 equiv.), T ₃ P (52.22 mg, 82.06 µmol , 48.81 µl, 50% pure, 1 eq.) and DIPEA (22.27 mg, 172.33 µmol, 30.02 µl, 2.1 eq.), and the mixture was heated at 25°C for 5 hours. 30 ml of a saturated aqueous sodium chloride solution was added and the mixture was extracted with ethyl acetate (30 ml×3) to give an organic phase. The organic phase was evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 42%-72%, 8 min) to give the compound WX022. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.77 (broad.d, J = 6.5 Hz, 1H), 8.32 (d, J = 2.0 Hz, 1H), 8.14 (s, 1H), 7.57 (d, J = 2.3 Hz, 1H), 7.41 - 7.31 (m, 2H), 7.20 - 7.07 (m, 2H), 5.45 (s, 2H), 5.30 - 5.08 (m, 1H), 4.76 (dd, J = 6.3, 9.5 Hz, 1H), 4.32 (t, J = 10.3 Hz, 1H), 3.93 - 3.87 (m, 1H); ¹⁹ F NMR (376 MHz, CDCl3) δ = -110.66 ~ -127.90 (m, 1F); LCMS m/z = 447.1[M+1] ⁺ .

Example 23. WX023

Synthesis path:

Step 1: Synthesis of Compound WX023-1

BB-4-3 (3.00 g, 10.09 mmol, 1 eq.) and N-bromosuccinimide (1.98 g, 11.10 mmol, 1.1 eq.) were slowly added to DCM (30 ml) at 25°C, and the reaction solution was continuously stirred at 25°C under nitrogen atmosphere for 12 hours. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with dichloromethane (100 ml×2). The organic phases were combined and washed successively with water (100 ml) and saturated aqueous sodium chloride solution (100 ml) to give the organic phase. The organic phase was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol=100:1~100:5) to give compound WX023-1.

Step 2: Synthesis of Compound WX023-2

In a round bottom flask was placed ethyl acetate (100 ml) at 25°C. Then WX023-1 (5.60 g, 14.89 mmol, 1 equiv.), T ₃ P (14.21 g, 22.33 mmol, 13.28 ml, 50% pure, 1.5 equiv.) and DIPEA (3.85 g, 29.77 mmol, 5.19 ml , 2 eq.). The reaction solution was continuously stirred at 25° C. under nitrogen for 1 hour. The reaction solution was slowly poured into water (200 ml), and the mixture was extracted with ethyl acetate (200 ml×2). The organic phases were combined and washed successively with water (200 ml) and saturated aqueous sodium chloride solution (200 ml) to give the organic phase. The organic phase was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:1) to give compound WX023-2.

Step 3: Synthesis of Compound WX023-3

In a round bottom flask was placed N,N-dimethylformamide (10 ml) at 25°C. Then WX023-2 (1.80 g, 5.03 mmol, 1 equiv.), iodomethane (5.71 g, 40.23 mmol, 2.50 ml, 8.01 equiv.) and cesium carbonate (3.60 g, 11.06 mmol, 2.2 equiv.) were slowly added. The reaction solution was continuously stirred at 25° C. under nitrogen for 1 hour. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined and washed successively with water (100 ml) and saturated aqueous sodium chloride solution (100 ml). The organic phase was evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~3:1) to give compound WX023-3. ¹ H NMR (400 MHz, _CDCl3 ) δ = 8.32 (d, J = 2.0 Hz, 1H), 7.61 (d, J = 2.3 Hz, 1H), 5.55 (broad s, J = 4.8 Hz, 1H), 4.79 - 4.44 (m, 2H), 4.34 - 4.11 (m, 1H), 3.46 (s, 3H), 1.40 (s, 9H).

Step 4: Synthesis of Compound WX023-4

Toluene (50 mL) was placed in a round bottom flask at 25°C, then WX023-3 (1.35 g, 3.63 mmol, 1 equiv.) and Lawsson's reagent (2.20 g, 5.44 mmol, 1.5 equiv.) were added slowly. The reaction solution was continuously stirred at 110° C. under nitrogen for 11 hours. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined and washed successively with water (100 ml) and saturated aqueous sodium chloride solution (100 ml). The organic phase was evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:1) to give compound WX023-4. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.38 (d, J = 2.0 Hz, 1H), 7.65 (d, J = 2.3 Hz, 1H), 6.10 (broad.d, J = 7.3 Hz, 1H), 4.81 - 4.67 (m, 1H), 4.58 (dd, J = 6.1, 9.4 Hz, 1H), 4.23 (dd, J = 9.5, 11.3 Hz, 1H), 3.87 (s, 3H), 1.41 (s, 9H) .

Step 5: Synthesis of Compound WX023-5

Dichloromethane (40 ml) was placed in a round bottom flask at 25°C, then WX023-4 (980 mg, 2.52 mmol, 1 equiv.) and trifluoroacetic acid (6.16 g, 54.02 mmol, 4, 21.40 equiv.) were added slowly. The reaction solution was continuously stirred at 25° C. under nitrogen for 11 hours. The reaction solution was slowly poured into a saturated sodium bicarbonate aqueous solution (100 ml), and the mixture was extracted twice with dichloromethane (100 ml×2). The organic phases were combined and washed successively with water (100 ml) and saturated aqueous sodium chloride solution (100 ml). The organic phase was evaporated under reduced pressure to give crude compound WX023-5, which was directly used in the next step without further purification. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.32 (d, J = 2.0 Hz, 1H), 7.59 (d, J = 2.3 Hz, 1H), 4.40 (dd, J = 6.3, 10.0 Hz, 1H), 4.10 (t, J = 10.8 Hz, 1H), 3.88 - 3.71 (m, 4H).

Step 6: Synthesis of Compound WX023

In a round bottom flask was placed N,N-dimethylacetamide (5 ml) at 25°C. Then WX023-5 (288 mg, 999.43 µmol, 1 equiv.), T ₃ P (954.00 mg, 1.50 mmol, 891.59 µl, 50% pure, 1.5 equiv.), BB-8 (406.16 mg, 2.00 mmol, 2 equiv.) and DIPEA (387.50 mg, 3.00 mmol, 522.24 µl, 3 equiv.). The reaction solution was continuously stirred at 25° C. under nitrogen for 2 hours. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined and washed successively with water (100 ml) and saturated aqueous sodium chloride solution (100 ml). The organic phase was evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 42%-72%, 8 min) to give the compound WX023. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.77 (broad.d, J = 7.0 Hz, 1H), 8.42 (d, J = 2.0 Hz, 1H), 8.04 (s, 1H), 7.71 (d, J = 2.0 Hz, 1H), 7.45 - 7.33 (m, 3H), 7.33 - 7.27 (m, 2H), 5.40 (s, 2H), 5.21 (td, J = 6.7, 11.3 Hz, 1H), 4.77 (dd, J = 6.3, 9.5 Hz, 1H), 4.33 (dd, J = 9.8, 11.0 Hz, 1H), 3.90 (s, 3H); LCMS m/z =475.0 [M+1] ⁺ .

Example 24. WX024

Synthesis path:

Step 1: Synthesis of Compound WX024-2

THF (100 ml) was placed in a round bottom flask. Then WX024-1 (10 g, 50.22 mmol, 1 eq.), BB-3-2 (11.34 g, 55.24 mmol, 1.1 eq.) and cesium carbonate (29.45 g, 90.39 mmol, 1.8 eq.) were slowly added. The mixture was continuously stirred at 65°C for 16 hours. The reaction solution was poured into water (300 ml), then a 2M hydrochloric acid solution was added to adjust the pH to 3. The mixture was extracted with ethyl acetate (500 ml×2). The organic phases were combined and washed with saturated aqueous sodium chloride solution (50 ml × 2). The organic phase was evaporated under reduced pressure to give compound WX024-2. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.46 (d, J = 1.8 Hz, 1H), 8.20 (dd, J = 2.0, 8.8 Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 5.62 (bd, J = 8.0 Hz, 1H), 4.83 - 4.61 (m, 2H), 4.45 (dd, J = 3.0, 9.3 Hz, 1H), 3.93 (s, 3H), 1.45 (s, 10H) .

Step 2: Synthesis of Compound WX024-3

MeOH (300 ml) was added to the hydrogenation flask. Then WX024-2 (25.8 g, 67.13 mmol, 1 equiv.) and Pd/C (2.6 g, 26.02 mmol, 10% purity) were slowly added. The atmosphere in the flask was changed to argon three times, and the mixture was continuously stirred at 30° C. under a hydrogen atmosphere (40 psi) for 24 hours. A new spot appeared on TLC (petroleum ether:ethyl acetate:acetic acid = 1:1:0.1). The reaction solution was filtered through Celite, and the filtrate was evaporated under reduced pressure to give compound WX024-3. ^1H NMR (400 MHz, CDCl ₃ ) δ = 7.50 - 7.44 (m, 1H), 7.39 (broad s, 1H), 6.70 (d, J = 8.5 Hz, 1H), 6.53 (broad s, 2H) , 6.00 (broad.d, J = 7.5 Hz, 1H), 4.73 (broad.d, J = 7.0 Hz, 1H), 4.43 (b.d., J = 8.5 Hz, 1H), 4.31 - 4.24 (m, 1H ), 3.85 (s, 3H), 1.41 (s, 9H).

Step 3: Synthesis of Compound WX024-4

Ethyl acetate (200 ml) was placed in a round bottom flask. Then WX024-3, T ₃ P (33.67 g, 52.91 mmol, 31.47 ml, 50% pure, 1.5 eq.) and DIEA (13.68 g, 105.83 mmol, 18.43 ml, 3 eq.) were slowly added and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was extracted with water (200 ml) and the layers were separated. The aqueous phase was extracted with ethyl acetate (200 ml x 2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (100 ml × 2) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:1) to give compound WX024-4. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.15 (broad s, 1H), 7.80 (dd, J = 2.0, 8.5 Hz, 1H), 7.73 (d, J = 2.0 Hz, 1H), 7.13 (d , J = 8.3 Hz, 1H), 5.62 (broad.d, J = 5.3 Hz, 1H), 4.71 - 4.59 (m, 2H), 4.33 - 4.23 (m, 1H), 3.95 - 3.89 (m, 3H), 1.43 (s, 9H).

Step 4: Synthesis of Compound WX024-5

N,N-dimethylformamide (200 ml) was placed in a round bottom flask. Then WX024-4 (17 g, 50.54 mmol, 1 eq.), cesium carbonate (49.40 g, 151.63 mmol, 3 eq.) and iodomethane (21.52 g, 151.63 mmol, 9.44 ml, 3 eq.) were added slowly, and the mixture stirred continuously at 25°C for 2 hours. Water (300 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (300 ml×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (100 ml × 2) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:1) to give compound WX024-5. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.92 - 7.83 (m, 2H), 7.18 (d, J = 8.4 Hz, 1H), 5.50 (bd, J = 6.1 Hz, 1H), 4.72 - 4.52 (m, 2H), 4.28 - 4.17 (m, 1H), 3.92 (s, 3H), 3.42 (s, 3H), 1.38 (s, 9H).

Step 5: Synthesis of Compound WX024-6

Toluene (40 ml) was placed in a round bottom flask. Then WX024-5 (4.01 g, 11.45 mmol, 1 equiv.), Lawsson's reagent (5.09 g, 12.59 mmol, 1.1 equiv.) and Boc ₂ O (10.60 g, 48.55 mmol, 11.15 ml, 4.24 equiv.) were added slowly. The mixture was heated to 110°C and stirred for 12 hours. The reaction solution was slowly poured into a saturated aqueous sodium chloride solution (30 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (30 ml×2) and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:1) to give compound WX024-6. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.99 - 7.92 (m, 2H), 7.21 (d, J = 8.3 Hz, 1H), 6.02 (broad.d, J = 7.8 Hz, 1H), 4.80 - 4.69 (m, 1H), 4.53 (dd, J = 6.3, 9.5 Hz, 1H), 4.22 (dd, J = 9.5, 11.3 Hz, 1H), 3.94 (s, 3H), 3.86 (s, 3H), 1.46 - 1.33 (m, 9H).

Step 6: Synthesis of Compound WX024-7

Ethyl acetate (10 ml) was placed in a round bottom flask. Then WX024-6 (500 mg, 1.36 mmol, 1 eq.) and HCl/ethyl acetate (4M, 10 ml, 29.31 eq.) were slowly added and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was evaporated under reduced pressure to give compound WX024-7. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.04 - 7.89 (m, 2H), 7.28 (br. s, 1H), 4.93 (br. s, 1H), 4.63 (br. s, 1H), 4.44 ( brs, 1H), 3.93 (s, 3H), 3.88 - 3.85 (m, 1H), 3.81 (s, 3H).

Step 7: Synthesis of Compound WX024-8

N,N-dimethylformamide (5 ml) was placed in a round bottom flask. Then WX024-7 (245 mg, 809.18 µmol, 1 eq, HCl), BB-8 (164.42 mg, 809.18 µmol, 1 eq), HATU (461.51 mg, 1.21 mmol, 1.5 eq) and DIPEA ( 313.74 mg, 2.43 mmol, 422.83 µl, 3 eq.), and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was poured into water (20 ml), and the mixture was extracted with ethyl acetate (20 ml×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (5 ml×2) and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:100) to give compound WX024-8. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.70 (broad.d, J = 7.3 Hz, 1H), 8.02 - 7.94 (m, 3H), 7.42 - 7.34 (m, 3H), 7.31 - 7.28 (m, 2H), 5.39 (s, 2H), 5.20 (td, J = 6.8, 11.0 Hz, 1H), 4.72 (dd, J = 6.5, 9.4 Hz, 1H), 4.31 (t, J = 10.3 Hz, 1H), 3.95 (s, 3H), 3.89 (s, 3H).

Step 8: Synthesis of Compound WX024

THF (4 ml) and H ₂ O (1 ml) were placed in a round bottom flask. Then WX024-8 (50 mg, 110.74 µmol, 1 eq.) and LiOH H ₂ O (4.65 mg, 110.74 µmol, 12.79 µl, 1 eq.) were slowly added and the mixture was continuously stirred at 25°C for 2 hours . The reaction solution was purified by HPLC (Column: Phenomenex Gemini-NX 80 mm*40 mm*3 µm; Mobile phase: [water (0.05% NH ₃ ⋅H ₂ O + 10 mM NH ₄ HCO ₃ )-ACN]); acetonitrile%: 11%-31%, 8 min) to give compound WX024. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.78 (br. s, 1H), 8.18 - 7.82 (m, 3H), 7.36 (br. s, 5H), 5.64 - 5.32 (m, 2H), 5.26 ( brs, 1H), 4.72 (brs, 1H), 4.32 (brs, 1H), 3.86 (brs, 3H); LCMS m/z = 438.1 [M+1] ⁺ .

Example 25. WX025

Synthesis path:

Step 1: Synthesis of Compound WX025-2

NaH (2.78 g, 69.38 mmol, 60% purity, 2.2 equiv.) was added to a solution of BB-3-2 (7.12 g, 34.69 mmol, 1.1 equiv.) in N,N-dimethylformamide (80 ml) at 0°C in nitrogen atmosphere, and the mixture was left to pass the reaction for 2 hours. The reaction solution was placed in an ice bath and WX025-1 (5 g, 31.54 mmol, 1 equiv.) was added. The reaction solution was heated to 25° C. and left to react for 12 hours. The mixture was slowly poured into ice water (100 ml) and extracted with ethyl acetate (150 ml x 2). The aqueous phase was adjusted to pH 5~6 with 1M HCl solution and extracted with ethyl acetate (150 ml×2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give compound WX025-2. LCMS m/z = 328.1 [M+1] ⁺ .

Stage 2: synthesis of compound WX025-3

Pd/C (3 g, 5% purity) was added to a solution of WX025-2 (9 g, 27.50 mmol, 1 eq.) in ethyl acetate (60 ml) and methanol (30 ml). The atmosphere in the flask was changed three times to H ₂ and the mixture was heated at 25° C. under a hydrogen atmosphere (15 psi) for 12 hours. The reaction solution was filtered through Celite, and the filtrate was evaporated under reduced pressure to give compound WX025-3. LCMS m/z = 298.1 [M+1] ⁺ .

Step 3: Synthesis of Compound WX025-4

T ₃ P (20.87 g, 32.79 mmol, 19.50 ml, 50% purity, 1.5 eq.) was added to a solution of DIPEA (7.06 g, 54.66 mmol, 9.52 ml, 2.5 eq.) and WX025-3 (6.5 g, 21.86 mmol, 1 eq.) in N,N-dimethylformamide (200 ml), and the mixture was heated at 25°C for 3 hours. Water (200 ml) was added to the reaction solution in an ice bath, and the mixture was extracted with ethyl acetate (200 ml×2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate=10:1~1:2) to give compound WX025-4. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.37 (br. s, 1 H), 8.20 - 8.31 (m, 2 H), 6.97 (d, J = 5.70 Hz, 1 H), 5.69 (br. d , J = 4.38 Hz, 1 H), 4.54 - 4.71 (m, 2 H), 4.22 - 4.35 (m, 1 H), 1.45 - 1.52 (m, 9 H); LCMS m/z = 280.1[M+1] ⁺ .

Step 4: Synthesis of Compound WX025-5

Iodomethane (384.21 mg, 2.71 mmol, 168.51 µl, 1.05 equiv.) was added to a solution of WX025-4 (0.72 g, 2.58 mmol, 1 equiv.) and cesium carbonate (1.01 g, 3.09 mmol, 1.2 equiv.) in tetrahydrofuran (14 ml), and the mixture was heated at 20°C for 12 hours. Water (25 ml) and ethyl acetate (25 ml) were added to the reaction solution, and the layers were separated. The aqueous phase was extracted with ethyl acetate (25 ml x 2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by column chromatography (dichloromethane:methanol=10:0~10:1) to give compound WX025-5. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.50 (s, 1H), 8.39 (d, J = 5.27 Hz, 1H), 7.06 (d, J = 5.40 Hz, 1H), 5.54 (broad. e, J = 6.02 Hz, 1 H), 4.56–4.73 (m, 2 H), 4.25–4.37 (m, 1 H), 3.47 (s, 3 H), 1.41 (s, 9 H); LCMS m/z = 294.1 [M+1] ⁺ .

Step 5: Synthesis of Compound WX025-6

Lawsson's reagent (110.32 mg, 272.74 µmol, 0.8 equiv.) was added to a solution of WX025-5 (0.1 g, 340.93 µmol, 1 equiv.) in toluene (2 ml) at 20°C, and the mixture was heated at 90°C for 12 hours. Ethyl acetate (5 ml) was added to the reaction solution. The mixture was filtered and the filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate=10:1~1:2) to give compound WX025-6. LCMS m/z = 310.1 [M+1] ⁺ .

Step 6: Synthesis of Compound WX025-7

HCl/ethyl acetate (3 ml) was added to a solution of WX025-6 (53 mg, 171.31 µmol, 1 eq.) in ethyl acetate (3 ml) and the mixture was heated at 25°C for 2 hours. The reaction solution was evaporated under reduced pressure to give compound WX025-7. LCMS m/z = 210.1 [M+1] ⁺ .

Step 7: Synthesis of Compound WX025

DIPEA (77.82 mg, 602.10 µmol, 104.87 µl, 3 equiv.) and T ₃ P (127.72 mg, 401.40 µmol, 119.36 µl, 2 equiv.) were added to WX025-7 solution (42 mg, 200.70 µmol, 1 equiv.) and BB-8 (61.17 mg, 301.05 µmol, 1.5 eq.) in N,N-dimethylformamide (3 ml) and the mixture was heated at 25°C for 2 hours. Water (5 ml) and ethyl acetate (5 ml) were added to the reaction solution, and the layers were separated. The aqueous phase was extracted with ethyl acetate (5 ml x 2). The organic phases were combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Phenomenex Luna C18 150 mm*30 mm*5 µm; Mobile phase: [water (0.05% HCl)-ACN]; acetonitrile%: 20%-50%, 12 min) to give the compound WX025. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.83 (d, J = 8.16 Hz, 2 H), 8.64 - 8.65 (m, 1 H), 8.67 (d, J = 7.72 Hz, 1 H), 8.54 (d, J = 5.28 Hz, 1 H), 7.26 - 7.42 (m, 6 H), 5.49 (s, 2 H), 5.04 (dt, J = 11.24, 6.84 Hz, 1 H), 4.44 - 4.64 ( m, 2 H), 3.83 (s, 3 H); LCMS m/z = 395.2 [M+1] ⁺ .

Example 26. WX026

Synthesis path:

Step 1: Synthesis of Compound WX026-2

Potassium carbonate (1.28 g, 9.26 mmol, 2 equiv.), copper bromide (66.40 mg, 462.89 µmol, 14.10 µl, 0.1 equiv.) and acetylacetone (23.17 mg, 231.45 µmol, 23.77 µl, 0.05 equiv.), and the mixture was stirred at 90°C for 10 hours. Water (100 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 ml×3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered, the filtrate was evaporated under reduced pressure to give compound WX026-2. LCMS m/z = 230.2 [M+1] ⁺ .

Stage 2: synthesis of compound WX026-3

To a solution of WX026-2 (0.45 g, 1.96 mmol, 1 equiv.) in tetrahydrofuran (5 ml) was added a solution of LiOH H ₂ O (164.76 mg, 3.93 mmol, 2 equiv.) in water (1 ml), and the mixture was stirred at 18°C for 1 hour. Ethyl acetate (20 ml) was added to the reaction solution, and the mixture was adjusted to pH 2 by adding 2N. hydrochloric acid solution. The organic phase was dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure to give compound WX026-3. LCMS m/z = 216.1 [M+1] ⁺ .

Step 3: Synthesis of Compound WX026

WX026-3 (153.52 mg, 713.37 µmol, 1.05 equiv.), DIPEA (263.42 mg, 2.04 mmol, 355.01 μl, 3 eq.) and T ₃ P (648.52 mg, 1.02 mmol, 606.09 μl, 50% pure, 1.5 eq.) at 18°C, and the mixture was stirred for 1 hour. The reaction solution was purified by HPLC (Column: Phenomenex Luna C18 150 mm*30 mm*5 µm; Mobile phase: [water (0.05% HCl)-ACN]; acetonitrile%: 20%-50%, 12 min) to give compound WX026 . ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.84 (s, 1H), 8.60 (d, J = 7.89 Hz, 1H), 8.36 (dd, J = 1.32, 4.82 Hz, 1H), 7.70 (dd , J = 1.53, 8.11 Hz, 1H), 7.25-7.46 (m, 5H), 5.49 (s, 2H), 4.85 (td, J = 7.67, 11.40 Hz, 1H), 4.63-4.75 (m, 1H), 4.51 (dd, J = 7.45, 9.65 Hz, 1H), 4.03 (broad s, 3H); LCMS m/z = 407.1 [M+1] ⁺ .

Example 27. WX027

Synthesis path:

Step 1: Synthesis of Compound WX027

In a round bottom flask was placed N,N-dimethylacetamide (5 ml) at 25°C. Then BB-4 (20 mg, 45.29 µmol, 1 equiv.), T ₃ P (43.23 mg, 67.94 µmol, 40.41 µl, 50% purity, 1.5 equiv.), BB-16 (15.85 mg, 67.94 µmol, 1.5 eq.) and DIPEA (17.56 mg, 135.88 μmol, 23.67 μl, 3 eq.), and the reaction solution was continuously stirred at 25°C under nitrogen atmosphere for 2 hours. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined, washed successively with water (100 ml) and saturated aqueous sodium chloride solution (100 ml), and evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 42%-72%, 8 min) to give the compound WX027. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.78 (broad.d, J = 7.0 Hz, 1H), 8.47 - 8.25 (m, 1H), 8.08 (s, 1H), 7.60 - 7.50 (m, 1H) , 7.35 (t, J = 7.9 Hz, 1H), 7.32 - 7.26 (m, 2H), 6.96 (t, J = 7.4 Hz, 1H), 6.91 (d, J = 8.0 Hz, 1H), 5.38 (s, 2H), 5.20 (td, J = 6.7, 11.4 Hz, 1H), 4.77 (dd, J = 6.3, 9.5 Hz, 1H), 4.38 - 4.23 (m, 1H), 3.92 (s, 3H), 3.84 (s , 3H); LCMS m/z =425.1 [M+1] ⁺ .

Example 28. WX028

Synthesis path:

Step 1: Synthesis of Compound WX028-2

MeCN (80 ml) was placed in a round bottom flask and then WX028-1 (4 g, 17.63 mmol, 1 eq.) was added slowly. After dissolution, 2-(2-bromoethoxy)tetrahydropyran (4.42 g, 21.16 mmol, 3.21 ml, 1.2 eq.) and DIPEA (2.51 g, 19.40 mmol, 3.38 ml, 1.1 eq.) were added, the mixture was heated to 90°C and continuously stirred for 3 hours. Water (100 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (30 ml×2) and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:1) to give compound WX028-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 4.55 (broad s, 1H), 4.34 (t, J = 5.3 Hz, 2H), 4.12 - 4.06 (m, 1H), 3.79 (td, J = 5.4, 10.8 Hz, 1H), 3.64 - 3.53 (m, 1H), 3.50 - 3.41 (m, 1H), 1.76 - 1.58 (m, 2H), 1.57 - 1.43 (m, 4H).

Stage 2: synthesis of compound WX028-3

THF (50 ml) was placed in a round bottom flask and then WX028-2 (4.99 g, 14.06 mmol, 1 eq.) was added slowly. The mixture was cooled to -78°C and n-butyl lithium (2.5 M, 6.75 ml, 1.2 eq.) was added. The mixture was stirred for 1 hour and benzaldehyde (2.98 g, 28.11 mmol, 2.84 ml, 2 eq.) was added slowly. The mixture was stirred at -78°C for 0.5 hours, then heated to -50°C and stirred for 0.5 hours. After completion of the reaction, the mixture was poured into a mixture of saturated aqueous ammonium chloride (200 ml) and ethyl acetate (200 ml), and the layers were separated. The aqueous phase was extracted with ethyl acetate (200 ml). The organic phases were combined, washed with saturated aqueous sodium chloride solution (50 ml×2) and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:1) to give compound WX028-3. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.45 - 7.29 (m, 5H), 6.09 (d, J = 5.8 Hz, 1H), 4.51 (broad s, 1H), 4.34 (ddq, J = 3.5 , 5.5, 7.2 Hz, 1H), 4.26 - 4.15 (m, 1H), 4.07 - 3.97 (m, 1H), 3.78 - 3.56 (m, 2H), 3.47 (dt, J = 5.9, 10.7 Hz, 1H), 1.85 - 1.64 (m, 2H), 1.52 (broad.d, J = 6.3 Hz, 4H).

Step 3: Synthesis of WX028-4 Compound

Toluene (50 ml) was placed in a round bottom flask. Then WX028-3 (4.7 g, 12.30 mmol, 1 eq.) and TsOH⋅H ₂ O (3.04 g, 15.98 mmol, 1.3 eq.) were slowly added and the mixture was continuously stirred at 110°C for 5 hours. Water (50 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 ml×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (20 ml × 2), and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:1) to give compound WX028-4. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.45 - 7.34 (m, 5H), 5.87 (s, 1H), 4.33 - 4.27 (m, 2H), 4.19 - 4.06 (m, 2H).

Step 4: Synthesis of Compound WX028-5

EtOH (20 ml) was placed into the autoclave. Then WX028-4 (500 mg, 1.78 mmol, 1 equiv.), Pd(dppf)Cl ₂ (261.21 mg, 356.99 µmol, 0.2 equiv.) and TEA (1.81 g, 17.85 mmol, 2.48 ml, 10 equiv.) were slowly added. ). The atmosphere in the autoclave was changed to argon three times, and the mixture was continuously stirred at 100° C. under CO (1 MPa) for 24 hours. Water (50 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 ml×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (10 ml×2) and evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 42%-72%, 8 min) to give the compound WX028-5. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.47 - 7.31 (m, 5H), 6.02 - 5.93 (m, 1H), 4.54 - 4.38 (m, 4H), 4.34 - 4.25 (m, 1H), 4.19 - 4.10 (m, 1H), 1.42 (t, J = 7.1 Hz, 3H).

Step 5: Synthesis of Compound WX028-6

THF (4 ml) and H ₂ O (1 ml) were placed in a round bottom flask. Then WX028-5 (260 mg, 951.38 µmol, 1 eq.) and LiOH⋅H ₂ O (119.77 mg, 2.85 mmol, 3 eq.) were added slowly. The mixture was continuously stirred at 25°C for 1 hour. The reaction solution was evaporated to dryness on a rotary evaporator under reduced pressure to remove THF. A 1M aqueous HCl solution was added dropwise to adjust pH to 3 and the mixture was filtered to give product WX028-6. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 7.40 (s, 5H), 6.00 (s, 1H), 4.50 - 4.40 (m, 1H), 4.40 - 4.29 (m, 2H), 4.26 - 4.11 ( m, 1H).

Step 6: Synthesis of Compound WX028-7

N,N-dimethylformamide (1 ml) was placed in a round bottom flask. Then WX028-6 (30 mg, 122.33 µmol, 1 eq.), BB-4 (54.02 mg, 122.33 µmol, 1 eq.), T ₃ P (116.77 mg, 183.50 µmol, 109.13 µl, 50% purity, 1.5 eq.) and DIPEA (47.43 mg, 366.99 μmol, 63.92 μl, 3 eq.), and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was purified by preparative HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 35%-65%, 8 min) to give a racemate compound WX028, which was then separated by chiral supercritical liquid chromatography (Column: DAICEL CHIRALCEL OD (250mm*30mm, 10µm); Mobile phase: [Neu-ETOH]; ethanol %: 45%-45%, min) to give the active isomer WX028 . ^1H NMR (400 MHz, _CDCl3 ) δ = 8.65 (broad.d, J = 7.1 Hz, 1H), 8.37 (b.d, J = 3.3 Hz, 1H), 7.56 (b.d, J = 7.8 Hz , 1H), 7.48 - 7.36 (m, 5H), 7.32 - 7.27 (m, 1H), 5.99 (s, 1H), 5.33 - 5.16 (m, 1H), 4.74 (dd, J = 6.4, 9.5 Hz, 1H ), 4.40 (broad.d, J = 4.3 Hz, 2H), 4.36 - 4.24 (m, 2H), 4.19 - 4.07 (m, 1H), 3.92 (s, 3H); LCMS m/z = 437.1 [M+1] ⁺ .

Example 29. WX029

Synthesis path:

Step 1: Synthesis of Compound WX029-2

MeCN (20 ml) was placed in a round bottom flask at 25°C, then WX029-1 (2.02 g, 15.89 mmol, 1 eq.), K ₂ CO ₃ (4.39 g, 31.79 mmol, 2 eq.) and 3- fluorobenzyl chloride (2.76 g, 19.07 mmol, 2.28 ml, 1.2 eq.). The reaction solution was continuously stirred at 60° C. under nitrogen for 12 hours. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined, washed successively with water (100 ml) and saturated aqueous sodium chloride solution (100 ml), and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:20~100:75) to give compound WX029-2. ^1H NMR (400 MHz, CDCl ₃ ) δ = 8.14 (s, 1H), 7.36 (dt, J = 5.9, 8.0 Hz, 1H), 7.11 - 7.03 (m, 2H), 6.99 (bd, J = 9.0 Hz, 1H), 5.42 (s, 2H), 4.14 - 3.89 (m, 3H).

Step 2: Synthesis of Compound WX029-3

THF (14 ml) and H ₂ O (2 ml) were placed in a round bottom flask at 25°C, and then WX029-2 and LiOH⋅H ₂ O (642.21 mg, 15.31 mmol, 3 eq.) were added slowly. The reaction solution was continuously stirred at 25° C. under nitrogen for 1 hour. The reaction solution was adjusted to pH 6~7 with dilute hydrochloric acid (1M), and the precipitate formed was filtered off and dried to give compound WX029-3.

Step 3: Synthesis of Compound WX029

N,N-dimethylacetamide (5 ml) was placed in a round bottom flask at 25°C, and then WX029-3 (15.03 mg, 67.94 µmol, 1.5 eq.), T ₃ P (43.23 mg, 67.94 µmol, 40.41 µl, 50% pure, 1.5 eq.), BB-4 (15.03 mg, 67.94 µmol, 1.5 eq.) and DIPEA (17.56 mg, 135.87 µmol, 23.67 µl, 3 eq.). The reaction solution was continuously stirred under nitrogen for 2 hours. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined, washed successively with water (100 ml) and saturated aqueous sodium chloride solution (100 ml), and evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 35%-65%, 8 min) to give the product WX029. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.83 (broad.d, J = 7.0 Hz, 1H), 8.37 (d, J = 3.8 Hz, 1H), 8.10 (s, 1H), 7.57 (d, J = 8.0 Hz, 1H), 7.40 - 7.27 (m, 2H), 7.15 - 6.87 (m, 3H), 5.40 (s, 2H), 5.29 - 5.07 (m, 1H), 4.79 (dd, J = 6.3, 9.3 Hz, 1H), 4.32 (t, J = 10.3 Hz, 1H), 3.93 (s, 3H); ¹⁹ F NMR (376 MHz, CDCl ₃ ) δ = -111.33 (s, 1F); LCMS m/z =413.0 [M+1] ⁺ .

Example 30. WX030

Synthesis path:

Step 1: Synthesis of Compound WX030-2

MeCN (20 ml) was placed in a round bottom flask at 25°C and then WX030-1 (2.04 g, 16.05 mmol, 1 eq.), K ₂ CO ₃ (4.44 g, 32.10 mmol, 2 eq.) and 4-methoxybenzyl chloride were added slowly (3.02 g, 19.26 mmol, 2.62 ml, 1.2 equiv.). The reaction solution was continuously stirred at 60° C. under nitrogen for 12 hours. The mixture was slowly poured into water (100 ml) and extracted with ethyl acetate (100 ml x 2). The organic phases were combined, washed successively with water (100 ml) and saturated aqueous sodium chloride solution (100 ml), evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:20~100:75) to give compound WX030-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.05 (s, 1H), 7.27 (d, J = 8.8 Hz, 2H), 6.98 - 6.87 (m, 2H), 5.43 - 5.26 (m, 2H), 4.03 - 3.97 (m, 3H), 3.82 (s, 3H).

Stage 2: synthesis of compound WX030-3

THF (14 ml) and H ₂ O (2 ml) were placed into a round bottom flask at 25°C, and WX030-2 (1.5 g, 6.07 mmol, 1 equiv.) and LiOH⋅H ₂ O (763.68 mg, 18.20 mmol, 3 equiv.). The mixture was continuously stirred at 25° C. under nitrogen for 1 hour. The reaction solution was adjusted to pH 6~7 with dilute hydrochloric acid (1 M), and the precipitate formed was filtered off and dried to give compound WX030-3.

Step 3: Synthesis of Compound WX030

N,N-dimethylacetamide (5 mL) was placed into a round bottom flask at 25°C, then WX030-3 (15.84 mg, 67.94 µmol, 1.5 equiv.), T ₃ P (43.23 mg, 67.94 µmol, 40.41 µL, 50 % purity, 1.5 equiv.), BB-4 (15.84 mg, 67.94 µmol, 1.5 equiv.) and DIPEA (17.56 mg, 135.87 µmol, 23.67 µl, 3 equiv.), and the solution was continuously stirred at 25°C under nitrogen atmosphere 2 hours. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined, washed successively with water (100 ml) and saturated aqueous sodium chloride solution (100 ml), evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 35%-65%, 8 min) to give the product WX030. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.80 (br.s, 1H), 8.37 (b.s, 1H), 7.98 (b.s, 1H), 7.56 (b.s, 1H), 7.38 - 7.28 (m, 1H), 7.26 (br.s, 2H), 6.91 (b.s, 2H), 5.41 - 5.10 (m, 3H), 4.78 (b.s, 1H), 4.31 (b.s, 1H ), 3.98 - 3.73 (m, 6H); LCMS m/z =425.1 [M+1] ⁺ .

Example 31. WX031

Synthesis path:

Step 1: Synthesis of Compound WX031-2

MeCN (20 ml) was placed in a round bottom flask. Then WX031-1 (1.5 g, 11.80 mmol, 1 eq.), K ₂ CO ₃ (2.45 g, 17.70 mmol, 1.5 eq.) and 4-fluorobenzyl chloride (1.88 g, 12.98 mmol, 1.55 ml, 1.1 eq.) were added. , and the mixture was continuously stirred at 50°C for 12 hours. The reaction solution was poured into water (50 ml), and the mixture was extracted with ethyl acetate (50 ml×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (10 ml×2) and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:100) to give compound WX031-2. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.09 (s, 1H), 7.30 (dd, J = 5.3, 8.3 Hz, 2H), 7.08 (t, J = 8.5 Hz, 2H), 5.39 (s, 2H ), 4.00 (s, 3H).

Step 2: Synthesis of Compound WX031-3

THF (8 ml) and H ₂ O (2 ml) were placed in a round bottom flask. Then WX031-2 (1.06 g, 4.52 mmol, 1 eq.) and LiOH⋅H ₂ O (569.16 mg, 13.56 mmol, 3 eq.) were added and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was evaporated under reduced pressure to remove THF. 1M hydrochloric acid solution was added dropwise to pH less than 3, and the mixture was filtered to give compound WX031-3. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.79 (s, 1H), 7.39 (dd, J = 5.5, 8.8 Hz, 2H), 7.25 - 7.17 (m, 2H), 5.47 (s, 2H) .

Step 3: Synthesis of Compound WX031

N,N-dimethylformamide (1 ml) was placed in a round bottom flask. Then WX031-3 (18.03 mg, 81.53 µmol, 1.2 eq.), BB-4 (30 mg, 67.94 µmol, 1 eq.), DIPEA (26.34 mg, 203.82 µmol, 35.50 µl, 3 eq.) and T ₃ P (64.85 mg, 101.91 µmol, 60.61 µl, 50% pure, 1.5 eq.), and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was directly purified by HPLC (Column: Xtimate C18 100*30mm*3µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 40%-70%, 8 min) to give compound WX031. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.81 (broad.d, J = 7.3 Hz, 1H), 8.37 (dd, J = 1.4, 4.6 Hz, 1H), 8.06 (s, 1H), 7.56 (dd , J = 1.4, 7.9 Hz, 1H), 7.33 - 7.27 (m, 3H), 7.07 (t, J = 8.5 Hz, 2H), 5.37 (s, 2H), 5.20 (td, J = 6.7, 11.2 Hz, 1H), 4.78 (dd, J = 6.4, 9.4 Hz, 1H), 4.32 (dd, J = 9.7, 10.9 Hz, 1H), 3.93 (s, 3H); LCMS m/z = 413.1 [M+1] ⁺ .

Example 32. WX032

Synthesis path:

Step 1: Synthesis of Compound WX032-2

WX032-1 (1.5 g, 11.80 mmol, 1 equiv.), 3-chloromethylbenzonitrile (1.97 g, 12.98 mmol, 1.72 ml, 1.1 equiv.) and K ₂ CO ₃ (2.45 g, 17.70 mmol , 1.5 eq.), and the reaction solution was continuously stirred at 50°C for 12 hours. Vodua (50 ml) was added to the solution and the mixture was extracted with ethyl acetate (50 ml x 2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (20 ml×2) and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:100) to give compound WX032-2. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.21 (s, 1H), 7.67 (t, J = 4.3 Hz, 1H), 7.58 (s, 1H), 7.52 (d, J = 4.8 Hz, 2H), 5.47 (s, 2H), 4.01 (s, 3H).

Stage 2: synthesis of compound WX032-3

THF (8 ml) and H ₂ O (2 ml) were placed in a round bottom flask. Then WX032-2 (1.4 g, 5.78 mmol, 1 eq.) and LiOH⋅H ₂ O (727.53 mg, 17.34 mmol, 3 eq.) were added, and the reaction solution was continuously stirred at 25°C for 2 hours. The solution was evaporated under reduced pressure to remove THF. 1M hydrochloric acid was added dropwise to pH less than 3, and the mixture was filtered to give the title compound WX032-3. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.83 - 8.80 (m, 1H), 7.86 - 7.80 (m, 2H), 7.68 - 7.57 (m, 2H), 5.59 - 5.52 (m, 2H).

Stage 3: Synthesis of compound WX032

N,N-dimethylformamide (1 ml) was placed in a round bottom flask. Then WX032-3 (15.50 mg, 67.94 µmol, 1 eq.), BB-4 (30 mg, 67.94 µmol, 1 eq.), DIPEA (26.34 mg, 203.82 µmol, 35.50 µl, 3 eq.) and T ₃ P (64.85 mg, 101.91 µmol, 60.61 µl, 50% pure, 1.5 eq.), and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was directly purified by HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 35%-65%, 8 min) to give the compound WX032. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.85 (broad.d, J = 7.3 Hz, 1H), 8.37 (d, J = 3.3 Hz, 1H), 8.17 (s, 1H), 7.65 (broad.d , J = 2.8 Hz, 1H), 7.60 - 7.55 (m, 2H), 7.53 - 7.47 (m, 2H), 7.29 (dd, J = 4.6, 7.9 Hz, 1H), 5.45 (s, 2H), 5.26 - 5.15 (m, 1H), 4.79 (dd, J = 6.3, 9.5 Hz, 1H), 4.38 - 4.28 (m, 1H), 3.93 (s, 3H); LCMS m/z = 420.1 [M+1] ⁺ .

Example 33. WX033

Synthesis path:

Step 1: Synthesis of Compound WX033-2

MeCN (20 ml) was placed in a round bottom flask. WX033-1 (1.5 g, 11.80 mmol, 1 eq.), 3-methylbenzyl chloride (1.83 g, 12.98 mmol, 1.72 ml, 1.1 eq.) and K ₂ CO ₃ (2.45 g, 17.70 mmol, 1.5 eq.) were added, and the mixture was continuously stirred at 50°C for 12 hours. Water (50 ml) was added to the solution and the mixture was extracted with ethyl acetate (50 ml x 2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (10 ml×2) and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:100) to give compound WX033-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.08 (s, 1H), 7.33 - 7.27 (m, 1H), 7.18 (d, J = 7.6 Hz, 1H), 7.10 (br.d, J = 1.8 Hz , 2H), 5.38 (s, 2H), 4.05 - 3.97 (m, 3H), 2.38 - 2.31 (m, 3H).

Stage 2: synthesis of compound WX033-3

THF (8 ml) and H ₂ O (2 ml) were placed in a round bottom flask. Then WX033-2 (1.6 g, 6.92 mmol, 1 eq.) and LiOH⋅H ₂ O (870.95 mg, 20.76 mmol, 3 eq.) were added and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was evaporated under reduced pressure to remove THF. 1M hydrochloric acid solution was added dropwise to pH less than 3, and the mixture was filtered to give compound WX033-3. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 13.28 (broad s, 1H), 8.79 (s, 1H), 7.30 - 7.23 (m, 1H), 7.17 - 7.07 (m, 3H), 5.43 ( s, 2H), 2.29 (s, 3H).

Step 3: Synthesis of Compound WX033

N,N-dimethylformamide (1 ml) was placed in a round bottom flask. Then WX033-3 (17.71 mg, 81.53 µmol, 1.2 eq.), BB-4 (30 mg, 67.94 µmol, 1 eq.), DIPEA (26.34 mg, 203.82 µmol, 35.50 µl, 3 eq.) and T ₃ P (64.85 mg, 101.91 µmol, 60.61 µl, 50% pure, 1.5 eq.) and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was directly purified by HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 37%-67%, 8 min) to give the compound WX033. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.81 (broad.d, J = 7.0 Hz, 1H), 8.37 (d, J = 3.3 Hz, 1H), 8.03 (s, 1H), 7.57 (d, J = 6.8 Hz, 1H), 7.31 - 7.27 (m, 2H), 7.20 - 7.15 (m, 1H), 7.11 - 7.07 (m, 2H), 5.36 (s, 2H), 5.22 (td, J = 6.8, 11.5 Hz, 1H), 4.79 (dd, J = 6.4, 9.4 Hz, 1H), 4.36 - 4.27 (m, 1H), 3.93 (s, 3H), 2.34 (s, 3H); LCMS m/z = 409.1 [M+1] ⁺ .

Example 34. WX034

Synthesis path:

Step 1: Synthesis of Compound WX034-2.

MeCN (20 ml) was placed in a single-necked flask. Then 1-chloromethyl-3,5-difluorobenzene (2.11 g, 12.98 mmol, 573.45 μl, 1.1 eq.), WX034-1 (1.5 g, 11.80 mmol, 1 eq.) and K ₂ CO ₃ (2.45 g, 17.70 mmol, 1.5 eq.), and the mixture was continuously stirred at 50°C for 12 hours. A new spot appeared on TLC (petroleum ether:ethyl acetate = 1:2). Water (50 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 ml×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (20 ml×2) and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:100) to give compound WX034-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.22 (s, 1H), 6.92 - 6.77 (m, 3H), 5.50 - 5.38 (m, 2H), 4.11 - 3.99 (m, 3H).

Stage 2: synthesis of compound WX034-3

THF (8 ml) and H ₂ O (2 ml) were placed in a round bottom flask. Then WX034-2 (1 g, 3.95 mmol, 1 eq.) and LiOH⋅H ₂ O (497.15 mg, 11.85 mmol, 3 eq.) were added and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was evaporated under reduced pressure to remove THF. 1M hydrochloric acid solution was added dropwise to pH less than 3 and the mixture was filtered to give compound WX034-3. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.80 (s, 1H), 7.24 (broad.t, J = 9.4 Hz, 1H), 7.07 (b.d, J = 6.3 Hz, 2H), 5.52 (s, 2H).

Step 3: Synthesis of Compound WX034.

N,N-dimethylformamide (1 ml) was placed in a round bottom flask. Then WX034-3 (19.50 mg, 81.53 µmol, 1.2 eq.), BB-4 (30 mg, 67.94 µmol, 1 eq.), DIPEA (26.34 mg, 203.82 µmol, 35.50 µl, 3 eq.) and T ₃ P (64.85 mg, 101.91 µmol, 60.61 µl, 50% pure, 1.5 eq.) and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was directly purified by HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 37%-67%, 8 min) to give the compound WX034. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.85 (broad.d, J = 7.0 Hz, 1H), 8.37 (dd, J = 1.5, 4.6 Hz, 1H), 8.16 (s, 1H), 7.57 (dd , J = 1.5, 8.0 Hz, 1H), 7.29 (dd, J = 4.7, 7.9 Hz, 1H), 6.85 - 6.73 (m, 3H), 5.38 (s, 2H), 5.21 (td, J = 6.6, 11.2 Hz, 1H), 4.79 (dd, J = 6.4, 9.5 Hz, 1H), 4.33 (dd, J = 9.6, 11.0 Hz, 1H), 3.93 (s, 3H); LCMS m/z = 431.1 [M+1] ⁺ .

Example 35. WX035

Synthesis path:

Step 1: Synthesis of Compound WX035-2

WX035-1 (1.5 g, 11.80 mmol, 1 equiv.), 1-chloromethyl-2-toluene (1.83 g, 12.98 mmol, 1.72 ml, 1.1 equiv.) and K ₂ CO ₃ (2.45 g, 17.70 mmol, 1.5 eq.), and the mixture was continuously stirred at 50°C for 12 hours. Water (50 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 ml×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (20 ml×2) and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:100) to give compound WX035-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.89 (s, 1H), 7.36 - 7.29 (m, 1H), 7.26 - 7.16 (m, 3H), 5.42 (s, 2H), 4.00 (s, 3H) , 2.27 (s, 3H).

Step 2: Synthesis of Compound WX035-3

THF (8 ml) and H ₂ O (2 ml) were placed in a round bottom flask. Then WX035-2 (1.6 g, 6.92 mmol, 1 eq.) and LiOH·H ₂ O (870.95 mg, 20.76 mmol, 3 eq.) were added and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was evaporated under reduced pressure to remove THF. 1M hydrochloric acid solution was added dropwise to pH less than 3 and the mixture was filtered to give WX035-3. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 13.28 (broad s, 1H), 8.79 (s, 1H), 7.30 - 7.23 (m, 1H), 7.17 - 7.07 (m, 3H), 5.43 ( s, 2H), 2.29 (s, 3H).

Step 3: Synthesis of Compound WX035

N,N-dimethylformamide (1 ml) was placed in a round bottom flask. Then WX035-3 (17.71 mg, 81.53 µmol, 1.2 eq.), BB-4 (30 mg, 67.94 µmol, 1 eq.), DIPEA (26.34 mg, 203.82 µmol, 35.50 µl, 3 eq.) and T ₃ P (64.85 mg, 101.91 µmol, 60.61 µl, 50% pure, 1.5 eq.) and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was directly purified by HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 37%-67%, 8 min) to give the compound WX035. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.81 (broad.d, J = 7.0 Hz, 1H), 8.37 (d, J = 4.5 Hz, 1H), 7.86 (s, 1H), 7.57 (d, J = 7.8 Hz, 1H), 7.34 - 7.27 (m, 2H), 7.26 - 7.15 (m, 3H), 5.40 (s, 2H), 5.22 (td, J = 6.7, 11.2 Hz, 1H), 4.79 (dd, J = 6.4, 9.4 Hz, 1H), 4.38 - 4.26 (m, 1H), 3.93 (s, 3H), 2.29 (s, 3H); LCMS m/z = 409.1 [M+1] ⁺ .

Example 36. WX036

Synthesis path:

Step 1: Synthesis of Compound WX036-2

In a round bottom flask was placed MeCN (20 ml) at 25°C. Then WX036-1 (2.02 g, 15.89 mmol, 1 equiv.), K ₂ CO ₃ (4.39 g, 31.79 mmol, 2 equiv.) and 1-chloro-3-chloromethylbenzene (3.07 g, 19.07 mmol, 2.42 ml , 1.2 equiv.). The reaction solution was continuously stirred at 60° C. under nitrogen for 12 hours. The mixture was slowly poured into water (100 ml) and extracted with ethyl acetate (100 ml x 2). The organic phases were combined, washed successively with water (100 ml) and saturated aqueous sodium chloride (100 ml) and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:20~100:75) to give compound WX036-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.15 (s, 1H), 7.35 - 7.26 (m, 3H), 7.17 (d, J = 6.8 Hz, 1H), 5.40 (s, 2H), 4.04 - 3.93 (m, 3H).

Step 2: Synthesis of Compound WX036-3

In a round bottom flask were placed THF (14 ml) and H ₂ O (2 ml) at 25°C. Then WX036-2 (1.80 g, 7.15 mmol, 1 equiv.) and LiOH⋅H ₂ O (900.33 mg, 21.46 mmol, 3 equiv.) were added slowly. The reaction solution was continuously stirred at 25° C. under nitrogen for 1 hour. The reaction solution was adjusted to pH 6~7 with dilute hydrochloric acid (1 M), and the precipitate was filtered off to give compound WX036-3.

Step 3: Synthesis of compound WX036.

In a round bottom flask was placed N,N-dimethylacetamide (5 ml) at 25°C. Then BB-4 (20 mg, 45.29 µmol, 1 equiv.), T ₃ P (43.23 mg, 67.94 µmol, 40.41 µl, 50% pure, 1.5 equiv.), WX036-3 (16.14 mg, 67.94 µmol, 1.5 eq.) and DIPEA (17.56 mg, 135.87 μmol, 23.67 μl, 3 eq.), and the reaction solution was continuously stirred at 25°C under nitrogen atmosphere for 2 hours. The reaction mixture was slowly poured into water (100 ml) and extracted with ethyl acetate (100 ml x 2). The organic phases were combined, washed successively with water (100 ml) and saturated aqueous sodium chloride (100 ml) and evaporated under reduced pressure to give the crude product. The crude product was isolated by preparative HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 35%-65%, 8 min) to give the compound WX036. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.95 - 8.69 (m, 1H), 8.37 (brd, J = 3.1 Hz, 1H), 8.10 (s, 1H), 7.68 - 7.47 (m, 1H) , 7.38 - 7.27 (m, 3H), 7.26 - 7.20 (m, 1H), 7.16 (bd, J = 7.0 Hz, 1H), 5.38 (s, 2H), 5.26 - 5.10 (m, 1H), 4.79 (dd, J = 6.5, 9.4 Hz, 1H), 4.41 - 4.20 (m, 1H), 3.93 (s, 3H); LCMS m/z =429.0 [M+1] ⁺ .

Example 37. WX037

Synthesis path:

Step 1: Synthesis of Compound WX037-2

In a round bottom flask was placed MeCN (20 ml) at 25°C. Then WX037-1 (2.03 g, 15.97 mmol, 1 equiv.), K ₂ CO ₃ (4.41 g, 31.94 mmol, 2 equiv.) and 1-bromo-3-chloromethylbenzene (3.94 g, 19.17 mmol, 241.82 μl , 1.2 equiv.). The reaction solution was continuously stirred at 60° C. under nitrogen for 12 hours. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined, washed successively with water (100 ml) and saturated aqueous sodium chloride (100 ml) and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:20~100:75) to give compound WX037-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.14 (s, 1H), 7.59 - 7.38 (m, 2H), 7.30 - 7.26 (m, 1H), 7.25 - 7.20 (m, 1H), 5.39 (s, 2H), 4.01 (s, 3H).

Stage 2: synthesis of compound WX037-3

In a round bottom flask were placed THF (14 ml) and H ₂ O (2 ml) at 25°C. Then WX037-2 (2.0 g, 6.75 mmol, 1 equiv.) and LiOH⋅H ₂ O (850.20 mg, 20.26 mmol, 3 equiv.) were added slowly. The reaction solution was continuously stirred at 25° C. under nitrogen for 1 hour. The reaction solution was adjusted to pH 6~7 with dilute hydrochloric acid (1 M), and the precipitate was filtered off to give compound WX037-3.

Step 3: Synthesis of Compound WX037

In a round bottom flask was placed N,N-dimethylacetamide (5 ml) at 25°C. Then BB-4 (20 mg, 45.29 µmol, 1 equiv.), T ₃ P (43.23 mg, 67.94 µmol, 40.41 µl, 50% purity, 1.5 equiv.), WX037-3 (19.16 mg, 67.94 µmol, 1.5 eq.) and DIPEA (17.56 mg, 135.87 μmol, 23.67 μl, 3 eq.), and the reaction solution was continuously stirred at 25°C under nitrogen atmosphere for 2 hours. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined, washed successively with water (100 ml) and saturated aqueous sodium chloride solution (100 ml), and evaporated under reduced pressure to give the crude product. The crude product was isolated by preparative HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 35%-65%, 8 min) to give the compound WX037. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.83 (b.d, J = 6.4 Hz, 1H), 8.37 (b.d, J = 4.5 Hz, 1H), 8.10 (s, 1H), 7.66 - 7.27 (m, 4H), 7.25 - 7.15 (m, 2H), 5.37 (s, 2H), 5.26 - 5.06 (m, 1H), 4.79 (broad dd, J = 6.5, 9.1 Hz, 1H), 4.33 (b .t, J = 10.3 Hz, 1H), 3.93 (s, 3H); LCMS m/z =474.8 [M+1] ⁺ .

Example 38. WX038

Synthesis path:

Step 1: Synthesis of Compound WX038-2

MeCN (20 ml) was placed in a round bottom flask. Then 1-chloromethyl-4-trifluoromethoxybenzene (2.73 g, 12.98 mmol, 517.03 µl, 1.1 eq.), WX038-1 (1.5 g, 11.80 mmol, 1 eq.) and K ₂ CO ₃ (2.45 g, 17.70 mmol, 1.5 eq.), and the mixture was continuously stirred at 50°C for 12 hours. Water (50 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 ml×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (20 ml×2) and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:100) to give compound WX038-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.17 (s, 1H), 7.36 (d, J = 8.5 Hz, 2H), 7.27 - 7.23 (m, 2H), 5.50 - 5.40 (m, 2H), 4.08 - 3.96 (m, 3H).

Step 2: Synthesis of WX038-3 Compound

THF (16 ml) and H ₂ O (4 ml) were placed in a round bottom flask. Then WX038-2 (1.9 g, 6.31 mmol, 1 eq.) and LiOH⋅H ₂ O (794.01 mg, 18.92 mmol, 3 eq.) were added and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was evaporated under reduced pressure to remove THF. 1M hydrochloric acid was added dropwise to pH less than 3, and the precipitate was filtered off to give compound WX038-3. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.81 (s, 1H), 7.51 - 7.33 (m, 4H), 5.53 (s, 2H).

Step 3: Synthesis of Compound WX038

N,N-dimethylformamide (1 ml) was placed in a round bottom flask. Then WX038-3 (23.41 mg, 81.53 µmol, 1.2 eq.), BB-4 (30 mg, 67.94 µmol, 1 eq.), DIPEA (26.34 mg, 203.82 µmol, 35.50 µl, 3 eq.) and T ₃ P (64.85 mg, 101.91 µmol, 60.61 µl, 50% pure, 1.5 eq.) and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was directly purified by HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 40%-70%, 8 min) to give the compound WX038. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.84 (broad d, J = 7.3 Hz, 1H), 8.38 (dd, J = 1.5, 4.5 Hz, 1H), 8.11 (s, 1H), 7.58 (dd , J = 1.5, 8.0 Hz, 1H), 7.34 (d, J = 8.8 Hz, 2H), 7.32 - 7.29 (m, 1H), 7.27 (s, 5H), 7.26 - 7.22 (m, 2H), 5.42 ( s, 2H), 5.28 - 5.15 (m, 1H), 4.80 (dd, J = 6.5, 9.5 Hz, 1H), 4.37 - 4.29 (m, 1H), 3.94 (s, 3H); LCMS m/z = 479.1 [M+1] ⁺ .

Example 39. WX039

Synthesis path:

Step 1: Synthesis of Compound WX039-2

In a round bottom flask was placed MeCN (20 ml) at 25°C. Then WX039-1 (2.0 g, 15.74 mmol, 1 equiv.), K ₂ CO ₃ (4.35 g, 31.48 mmol, 2 equiv.) and 1-chloromethyl-4-methylbenzene (2.66 g, 18.89 mmol, 2.48 ml , 1.2 equiv.). The reaction solution was continuously stirred at 60° C. under nitrogen for 12 hours. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined, washed successively with water (100 ml) and saturated aqueous sodium chloride (100 ml) and evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:20~100:75) to give compound WX039-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.04 (s, 1H), 7.23 - 7.17 (m, 4H), 5.37 (s, 2H), 4.00 (s, 3H), 2.47 - 2.25 (m, 3H) .

Step 2: Synthesis of Compound WX039-3

In a round bottom flask were placed THF (14 ml) and H ₂ O (2 ml) at 25°C. Then WX039-2 (1.5 g, 6.49 mmol, 1 equiv.) and LiOH⋅H ₂ O (816.52 mg, 19.46 mmol, 3 equiv.) were added slowly. The reaction solution was continuously stirred at 25° C. under nitrogen for 1 hour. The reaction solution was adjusted to pH 6~7 with dilute hydrochloric acid (1 M), and the precipitate was filtered off to give compound WX039-3.

Step 3: Synthesis of Compound WX039

BB-4 (20 mg, 45.29 µmol, 1 equiv.), T ₃ P (43.23 mg, 67.94 µmol, 40.41 µl, 50% purity, 1.5 equiv.), WX039- 3 (14.76 mg, 67.94 µmol, 1.5 eq.) and DIPEA (17.56 mg, 135.87 µmol, 23.67 µl, 3 eq.) at 25°C, and the reaction solution was continuously stirred at 25°C under nitrogen atmosphere for 2 hours. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined, washed successively with water (100 ml) and saturated aqueous sodium chloride (100 ml) and evaporated under reduced pressure to give the crude product. The crude product was isolated by preparative HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 40%-70%, 8 min) to give the product WX039. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.80 (broad.d, J = 7.3 Hz, 1H), 8.37 (dd, J = 1.5, 4.5 Hz, 1H), 8.00 (s, 1H), 7.56 (dd , J = 1.5, 7.8 Hz, 1H), 7.28 (dd, J = 4.8, 8.0 Hz, 1H), 7.19 (s, 4H), 5.35 (s, 2H), 5.21 (td, J = 6.8, 11.0 Hz, 1H), 4.78 (dd, J = 6.4, 9.4 Hz, 1H), 4.32 (dd, J = 9.7, 11.2 Hz, 1H), 3.93 (s, 3H), 2.63–2.13 (m, 3H); LCMS m/z =409.1 [M+1] ⁺ .

Example 40. WX040

Synthesis path:

Step 1: Synthesis of WX040-2 Compound

THF (100 ml) was placed in a round bottom flask. Then WX040-1 (10 g, 50.22 mmol, 1 eq.), BB-3-2 (11.34 g, 55.24 mmol, 1.1 eq.) and cesium carbonate (29.45 g, 90.39 mmol, 1.8 eq.) were slowly added, and the mixture was continuously stirred at 65°C for 16 hours. TLC monitoring (petroleum ether:ethyl acetate:acetic acid = 1:1:0.1) showed that the starting compounds had completely reacted. The reaction solution was poured into water (300 ml), then a 2M hydrochloric acid solution was added to adjust the pH to 3. The mixture was extracted with ethyl acetate (500 ml×2). The organic phases were combined and washed with saturated aqueous sodium chloride solution (50 ml x 2). The organic phase was then evaporated under reduced pressure to give WX040-2. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.46 (d, J = 1.8 Hz, 1H), 8.20 (dd, J = 2.0, 8.8 Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 5.62 (bd, J = 8.0 Hz, 1H), 4.83 - 4.61 (m, 2H), 4.45 (dd, J = 3.0, 9.3 Hz, 1H), 3.93 (s, 3H), 1.45 (s, 10H) .

Stage 2: synthesis of compound WX040-3

MeOH (300 ml) was placed in a hydrogenation flask. Then WX040-2 (25.8 g, 67.13 mmol, 1 equiv.) and Pd/C (2.6 g, 26.02 mmol, 10% purity) were added slowly. The atmosphere in the flask was changed to argon three times and the mixture was continuously stirred at 30° C. under a hydrogen atmosphere (40 psi) for 24 hours. A new spot appeared on TLC (petroleum ether:ethyl acetate:acetic acid = 1:1:0.1). The reaction solution was filtered through Celite, and the filtrate was evaporated under reduced pressure to give compound WX040-3. ^1H NMR (400 MHz, CDCl ₃ ) δ = 7.50 - 7.44 (m, 1H), 7.39 (broad s, 1H), 6.70 (d, J = 8.5 Hz, 1H), 6.53 (broad s, 2H) , 6.00 (broad.d, J = 7.5 Hz, 1H), 4.73 (broad.d, J = 7.0 Hz, 1H), 4.43 (b.d., J = 8.5 Hz, 1H), 4.31 - 4.24 (m, 1H ), 3.85 (s, 3H), 1.41 (s, 9H).

Step 3: Synthesis of Compound WX040-4

Ethyl acetate (200 ml) was placed in a round bottom flask. Then WX040-3 (12.5 g, 35.28 mmol, 1 eq.), T ₃ P (33.67 g, 52.91 mmol, 31.47 ml, 50% pure, 1.5 eq.) and DIPEA (13.68 g, 105.83 mmol, 18.43 ml) were slowly added. , 3 eq.), and the mixture was continuously stirred at 25°C for 2 hours. Water (200 ml) was added to the reaction solution, the layers were separated. The aqueous phase was extracted with ethyl acetate (200 ml x 2). The organic phases were combined and washed with saturated aqueous sodium chloride solution (100 ml×2), then the organic phase was evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:1) to give compound WX040-4. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.15 (broad s, 1H), 7.80 (dd, J = 2.0, 8.5 Hz, 1H), 7.73 (d, J = 2.0 Hz, 1H), 7.13 (d , J = 8.3 Hz, 1H), 5.62 (broad.d, J = 5.3 Hz, 1H), 4.71 - 4.59 (m, 2H), 4.33 - 4.23 (m, 1H), 3.95 - 3.89 (m, 3H), 1.43 (s, 9H).

Step 4: Synthesis of Compound WX040-5

N,N-dimethylformamide (200 ml) was placed in a round bottom flask. then WX040-4 (17 g, 50.54 mmol, 1 eq.), cesium carbonate (49.40 g, 151.63 mmol, 3 eq.) and iodomethane (21.52 g, 151.63 mmol, 9.44 ml, 3 eq.) were slowly added, and the mixture stirred continuously at 25°C for 2 hours. Water (300 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (300 ml×2). The organic phases were combined and washed with saturated aqueous sodium chloride solution (100 ml×2), the organic phase was then evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:1) to give compound WX040-5. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.92 - 7.83 (m, 2H), 7.18 (d, J = 8.4 Hz, 1H), 5.50 (bd, J = 6.1 Hz, 1H), 4.72 - 4.52 (m, 2H), 4.28 - 4.17 (m, 1H), 3.92 (s, 3H), 3.42 (s, 3H), 1.38 (s, 9H).

Step 5: Synthesis of Compound WX040-6

Toluene (40 ml) was slowly added WX040-5 (4.01 g, 11.45 mmol, 1 equiv.), Lawsson's reagent (5.09 g, 12.59 mmol, 1.1 equiv.) and Boc ₂ O (10.60 g, 48.55 mmol, 11.15 ml, 4.24 eq.), and the mixture was heated to 110°C and continuously stirred for 12 hours. The reaction solution was slowly poured into a saturated aqueous sodium chloride solution (30 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined and washed with saturated aqueous sodium chloride solution (30 ml×2), the organic phase was then evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1~1:1) to give compound WX040-6. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.99 - 7.92 (m, 2H), 7.21 (d, J = 8.3 Hz, 1H), 6.02 (br.d, J = 7.8 Hz, 1H), 4.80 - 4.69 (m, 1H), 4.53 (dd, J = 6.3, 9.5 Hz, 1H), 4.22 (dd, J = 9.5, 11.3 Hz, 1H), 3.94 (s, 3H), 3.86 (s, 3H), 1.46 - 1.33 (m, 9H).

Step 6: Synthesis of Compound WX040-7

THF (8 ml) and H ₂ O (2 ml) were placed in a round bottom flask. Then WX040-6 (800 mg, 2.18 mmol, 1 eq.) and LiOH⋅H ₂ O (183.23 mg, 4.37 mmol, 2 eq.) were slowly added and the mixture was continuously stirred at 25°C for 2 hours. Dilute hydrochloric acid was added dropwise to the reaction solution until pH<3, and the precipitate was filtered off to give compound WX040-7. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.89 - 7.79 (m, 1H), 7.25 - 7.20 (m, 1H), 7.02 - 6.95 (m, 1H), 6.17 - 6.02 (m, 1H), 4.88 - 4.74 (m, 1H), 4.56 (dd, J = 6.4, 9.7 Hz, 1H), 4.31 - 4.17 (m, 1H), 3.85 (s, 3H), 1.41 (s, 9H).

Step 7: Synthesis of Compound WX040-8

N,N-dimethylformamide (5 ml) was placed in a round bottom flask. Then WX040-7 (200 mg, 567.53 µmol, 1 equiv.), NH ₄ Cl (91.07 mg, 1.70 mmol, 3 equiv.), HATU (647.37 mg, 1.70 mmol, 253.15 µl, 3.00 equiv.) and DIPEA were slowly added. (366.75 mg, 2.84 mmol, 494.27 µl, 5 eq.) and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was directly purified by HPLC (Column: Xtimate C18 100 mm*30 mm*3 µm; Mobile phase: [water (0.225% formic acid)-ACN]; acetonitrile%: 30%-60%, 8 min) to give the compound WX040-8. ^1H NMR (400 MHz, _CDCl3 ) δ = 7.82 (s, 1H), 7.64 (bd, J = 8.3 Hz, 1H), 7.22 (d, J = 8.3 Hz, 1H), 6.98 (s, 2H ), 5.01 (broad s, 1H), 4.86 - 4.66 (m, 1H), 4.52 (dd, J = 6.3, 9.5 Hz, 1H), 4.30 - 4.11 (m, 1H), 3.86 (s, 3H), 1.40 (s, 9H).

Step 8: Synthesis of Compound WX040-9

Ethyl acetate (10 ml) was placed in a round bottom flask. Then WX040-8 (40 mg, 113.82 µmol, 1 eq.) and HCl/ethyl acetate (4M, 10.00 ml, 351.42 eq.) were slowly added and the mixture was continuously stirred at 25°C for 1 hour. The reaction solution was evaporated under reduced pressure to give compound WX040-9. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 8.16 - 8.04 (m, 2H), 7.91 (d, J = 8.5 Hz, 1H), 7.57 (s, 1H), 7.37 (d, J = 8.3 Hz , 1H), 4.62 - 4.41 (m, 3H), 3.82 (s, 3H).

Step 9: Synthesis of Compound WX040

WX040-9 (30 mg, 119.38 µmol, 1 equiv., HCl), BB-8 (24.26 mg, 119.38 µmol, 1 equiv.), HATU (68.09 mg, 179.07 µmol, 1.5 equiv.) .) and DIEA (46.28 mg, 358.13 µmol, 62.38 µl, 3 eq.), and the mixture was continuously stirred at 25°C for 2 hours. The reaction solution was directly purified by HPLC (Column: Phenomenex Gemini-NX 80 mm*30 mm*3 µm; Mobile phase: [water (10 mM NH ₄ HCO ₃ )-ACN]; acetonitrile%: 28%-58%, 9 min ) to get a WX040 connection. ¹ H NMR (400 MHz, CDCl ₃ ) δ =8.70 (broad.d, J = 7.3 Hz, 1H), 8.04 (s, 1H), 7.69 (s, 1H), 7.39 - 7.36 (m, 1H), 7.31 - 7.28 (m, 7H), 5.39 (s, 2H), 5.20 (td, J = 6.8, 11.0 Hz, 1H), 4.72 (dd, J = 6.5, 9.4 Hz, 1H), 4.31 (t, J = 10.3 Hz, 1H), 3.89 (s, 3H); LCMS m/z = 437.1 [M+1] ⁺ .

Example 41. WX041

Synthesis path:

Step 1: Synthesis of Compound WX041-2

In a round bottom flask was placed MeCN (20 ml) at 25°C. Then WX041-1 (2.68 g, 21.09 mmol, 1 equiv.), K ₂ CO ₃ (5.83 g, 42.17 mmol, 2 equiv.) and 4-(chloromethyl)-trifluoromethylbenzene (4.92 g, 25.30 mmol, 3.73 ml) were slowly added , 1.2 equiv.). The reaction solution was continuously stirred at 60° C. under nitrogen for 12 hours. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined and washed successively with water (100 ml) and saturated aqueous sodium chloride solution (100 ml), the organic phase was then evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:20-100:75) to give compound WX041-2. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.17 (s, 1H), 7.65 (d, J = 8.0 Hz, 2H), 7.40 (d, J = 8.0 Hz, 2H), 5.49 (s, 2H), 4.08 - 3.91 (m, 3H).

Step 2: Synthesis of Compound WX041-3

THF (20 ml) and H ₂ O (4 ml) were placed in a round bottom flask at 25°C. WX041-2 (2.40 g, 8.41 mmol, 1 equiv.) and LiOH⋅H ₂ O (1.06 g, 25.24 mmol, 3 equiv.) were added slowly. The reaction solution was continuously stirred at 25° C. under nitrogen for 1 hour. The reaction solution was adjusted to pH 6~7 with dilute hydrochloric acid (1 M), and the precipitate was filtered off to give compound WX041-3.

Step 3: Synthesis of Compound WX041

In a round bottom flask was placed N,N-dimethylacetamide (5 ml) at 25°C. BB-4 (20 mg, 45.29 µmol, 1 equiv.), T ₃ P (43.23 mg, 67.94 µmol, 40.41 µl, 50% purity, 1.5 equiv.), WX041-3 (18.42 mg, 67.94 µmol, 1.5 equiv.) and DIPEA (17.56 mg, 135.87 μmol, 23.67 μl, 3 equiv.), and the reaction solution was continuously stirred at 25°C under nitrogen atmosphere for 2 hours. The reaction solution was slowly poured into water (100 ml), and the mixture was extracted with ethyl acetate (100 ml×2). The organic phases were combined and washed successively with water (100 ml) and saturated aqueous sodium chloride solution (100 ml), then the organic phase was evaporated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (Column: Phenomenex Gemini-NX 80 mm*30 mm*3 µm; Mobile phase: [water (10 mM NH ₄ HCO ₃ )-ACN] to give compound WX041. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.77 (brd, J = 7.5 Hz, 1H), 8.31 (brd, J = 3.6 Hz, 1H), 8.07 (s, 1H), 7.65 - 7.45 (m, 3H), 7.33 (broad d, J = 8.0 Hz, 2H), 7.22 (dd, J = 4.8, 7.9 Hz, 1H), 5.40 (s, 2H), 5.23 - 4.97 (m, 1H), 4.72 (dd, J = 6.5 , 9.4 Hz, 1H), 4.25 (t, J = 10.3 Hz, 1H), 3.87 (s, 3H), ¹⁹ F NMR (377 MHz, CDCl ₃ ) δ = -43.60 (s, 1F), LCMS m/z =463.1 [M+1] ⁺ .

Example 42. WX042

Synthesis path:

Step 1: Synthesis of Compound WX042-2

WX042-1 (2 g, 15.74 mmol, 1 equiv.), 3-chloromethylanisole (2.46 g, 15.74 mmol, 2.14 ml, 1 equiv.) and K ₂ CO ₃ (2.61 g, 18.88 mmol , 1.2 eq.), and the mixture was heated at 50°C for 12 hours. The reaction solution was filtered to remove K ₂ CO ₃ . The filter cake was washed with ethyl acetate (15 ml×3) and the filtrate was washed with saturated aqueous sodium chloride (50 ml). The organic phase was evaporated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1-100:100) to give compound WX042-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.10 - 8.01 (m, 1H), 7.30 - 7.21 (m, 1H), 6.91 - 6.80 (m, 2H), 6.77 (s, 1H), 5.34 (s, 2H), 3.96 (s, 3H), 3.75 (s, 3H).

Stage 2: synthesis of compound WX042-3

WX042-2 (2.5 g, 10.11 mmol, 1 equiv.) was added to THF (30 ml). Then a solution of LiOH⋅H ₂ O (509.17 mg, 12.13 mmol, 1.2 equiv.) in H ₂ O (3 ml) was added. The reaction mixture was stirred at 25°C for 3 hours. TLC monitoring (petroleum ether:ethyl acetate = 1:1) showed that the starting compounds had completely reacted. The reaction solution was adjusted to pH 6 by adding 1M hydrochloric acid, and the mixture was evaporated under reduced pressure to remove the solvent. Toluene (10 ml) was added and the mixture was evaporated under reduced pressure once more to remove toluene to give compound WX042-3. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.19 (s, 4H), 7.21 - 7.17 (m, 1H), 4.65 (s, 2H), 3.79 (s, 3H); LCMS m/z = 233.9 [M+1] ⁺ .

Stage 3: Synthesis of compound WX042

BB-4 (94.67 mg, 214.39 µmol, 1 equiv.), WX042-3 (50 mg, 214.39 µmol, 1 equiv.), T ₃ P (163.71 mg, 257.26 µmol , 153.00 µl, 50% pure, 1.2 eq.) and DIPEA (41.56 mg, 321.58 µmol, 56.01 µl, 1.5 eq.), and the mixture was heated at 25°C for 2 hours. LCMS analysis showed that the parent compounds had completely reacted. The reaction solution was evaporated under reduced pressure to a volume of about 5 ml. The residue was purified by HPLC (formic acid system) to give compound WX042. ^1H NMR (400 MHz, _CDCl3 ) δ = 8.81 (broad.d, J = 7.0 Hz, 1H), 8.37 (dd, J = 1.6, 4.6 Hz, 1H), 8.04 (s, 1H), 7.57 (dd , J = 1.6, 7.9 Hz, 1H), 7.33 - 7.27 (m, 2H), 7.00 - 6.69 (m, 3H), 5.36 (s, 2H), 5.22 (td, J = 6.7, 11.1 Hz, 1H), 4.79 (dd, J = 6.3, 9.5 Hz, 1H), 4.32 (dd, J = 9.5, 11.0 Hz, 1H), 3.93 (s, 3H), 3.79 (s, 3H).

Test Example 1: In Vitro Testing

Testing stages:

1.1 Reagents

Reagent Brand Batch number RPMI-1640 GIBCO 11875-119 10XDPBS GIBCO 14200-075 Fetal bovine serum (FBS) biowest S1810-500 Penicillin/streptomycin (100X) biowest L0022-100 CellTiter Glo Promega G7571 Q-VD-OPh Hydrate Sigma SML0063-5MG Recombinant human TNF-a Pepotech 300-01A-50

1.2 Instruments and materials

CULTURPLATE-384 +LID /50W Brand PerkinElmer cat#6007680

Envision tablet reader (Perkin Elmer, Cat#2104-0010)

1.3 Preparation of reagents

(1) 1×DPSB: 900 ml of deionized water was added to 100 ml of 10×DPBS.

(2) 1640 complete medium: To 89 ml of RPMI-1640 medium was added 10 ml of FBS and 1 ml of penicillin/streptomycin

2. Stages of testing

1. U937 cells were centrifuged and then the medium was discarded. Cells were resuspended in complete mixture 1640 and counted. The cell concentration was adjusted to 5×10 ⁵ cells/ml for use, and the cells were distributed according to the test compounds.

2. QVD-OPh at a final concentration of 25 μM and TNF-α at a final concentration of 100 ng/ml were added to the resulting U937 cell suspension, after which the mixture was gently mixed.

3. 30 µl/well of U937 cells containing 25 µM QVD-OPh and TNF-α at a final concentration of 100 ng/ml was added to a 384-well plate, and blank control wells without QVD-OPh and TNF-α were prepared.

4. To the compound addition group, 10 μl of test compound was added at 4× the final concentration, and DMSO at the appropriate concentration was added to the blank control wells.

5. The 384-well plate was placed in an incubator with 37°C and 5% CO ₂ for 24 hours.

6. After 24 hours, the 384-well plate was removed and equilibrated at room temperature for 20 minutes.

7. 40 μl of CTG was added to each well, and then the plate was shaken on a shaker in the dark for 2 minutes.

8. The plate was incubated at room temperature in the dark for 10 minutes and then read on an Envision plate reader.

Test results:

The results of testing the compounds of the present invention are presented in table 1.

Table 1. Results of testing the activity of the compounds of the present invention on U937 cells Compound IC ₅₀ in cell assay (nM) Compound IC ₅₀ in cell assay (nM) WX001 0.5 WX023 29 WX002 1.8 WX024 43 WX003 0.5 WX025 12 WX004 0.3 WX026 5 WX005 2 WX027 20 WX006 9 WX028 4.7 WX007 70 WX029 2.2 WX008 8.1 WX030 15.6 WX009 1.4 WX031 2.4 WX010 47 WX032 5.7 WX011 1.7 WX033 7.6 WX012 12.1 WX034 3.7 WX013 1.8 WX035 4.9 WX014 3.7 WX036 6.4 WX015 5.1 WX037 24 WX016 9.8 WX038 238 WX017 59 WX039 3.4 WX018 1.3 WX040 4.8 WX019 0.6 WX041 86.5 WX020 0.6 WX042 24.8 WX021 13 WX022 20

Conclusion on the results of testing

The results shown in the table above show that the compounds of the present invention exhibit good inhibitory activity in the TNFα/QVD-OPh-induced programmed cell death (necroptosis) inhibition activity cellular assay.

Test Example 2: In Vitro Activity Testing on Mouse RIP1

cell line

L929 (purchased from ATCC, Item No.: CCL-1)

Reagent

Reagent Name Brand Number EMEM environment ATCC 30-2003 Horse serum Gibco 16050130 Dual antibiotic mix (penicillin, streptomycin) Millipore TMS-AB2-C Recombinant mouse tumor necrosis factor alpha (TNF-α) R&D 410-MT-010 Caspase inhibitor (QVD-OPh) R&D OPH001-01M CellTiter Glo fluorescent cells Promega G7573

device

Tablet reader (Perkin Elmer, Envision)

Preparation of reagents

L929 cell complete medium: EMEM + 10% horse serum (v/v ratio) + 1% dual antibiotic mix (v/v ratio)

Testing stages

1. Grown L929 cells were trypsinized, centrifuged, resuspended in complete growth medium, and then counted. The cell concentration was adjusted to 1×10 ⁵ /ml by diluting with complete growth medium.

2. The cell suspension was added to a 96-well plate at a concentration of 100 μl per well and incubated at 37°C and 5% CO ₂ for 24 hours.

3. Different concentrations of test compounds were added to each well, and dimethyl sulfoxide was added to the control group. The tablet was incubated at 37°C and 5% CO ₂ for 1 hour.

4. TNF-α at a final concentration of 50 ng/ml and Q-VD at a final concentration of 50 μM were added to each well, and the plate was incubated at 37°C and 5% CO ₂ for 24 hours.

5. 50 µl CellTiter Glo fluorescent cells were added to each well and the plate was shaken in the dark for 10 minutes.

6. The tablet was read on a tablet reader.

Test results

The results of testing the compounds of the present invention are presented in table 2.

Table 2. Results of testing the activity of the compounds of the present invention on L929 cells Compound IC ₅₀ in cell assay (nM) WX001 1161 WX003 1213 WX005 1311 WX008 47 WX009 103 WX011 eleven WX013 14 WX019 51 WX020 29 WX021 445 WX028 65 WX029 184 WX031 174 WX032 255 WX039 64

Conclusion on the results of testing

The results shown in the table above show that the compounds of the present invention exhibit good inhibitory activity in a cellular assay of TNFα/QVD-OPh-induced programmed cell death (necroptosis) inhibition activity in mouse L929 cells.

Test Example 3: Compound Pharmacokinetic Study

Testing stages

Eight healthy adult C57BL/6 mice (6-9 weeks old, purchased from Shanghai Lingchang Biotechnology Co., Ltd.) were selected and randomly assigned to two groups of 4 mice per group (n=2). One group was administered 0.5 mg/kg of the test compound via intravenous injection, and the other group was administered 1 mg/kg of the test compound via a force-feeding tube. Blood samples were taken, respectively, from animals from the "intravenous" group and from the "oral" group after 0.083, 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 24 hours after administration. The concentration of the compound in the blood was determined using the LC-MS/MS method. Pharmacokinetic parameters were calculated with WinNonlin™ Version 6.3 software (Pharsight, Mountain View, CA) using the non-compartmental method and the linear logarithmic trapezoid method. AUC _0-last is the area under the plasma concentration-time curve from point 0 to the time of the last detectable concentration; C _max is the peak concentration; T _max is the time to peak; T _1/2 is the half-life; CL is the hatch rate; T _last is the time corresponding to the last quantifiable concentration; po means "by mouth"; iv means intravenous injection; and F% is oral boavailability.

Test results

The test results for the compound of the present invention are shown in Table 3.

Table 3. Pharmacokinetic characteristics of the compound Compound WX009 i.v. T _1/2 (hour) 1.4 Vd _ss (l/kg) 1.1 CL (ml/min/kg) 9.4 AUC _0-last (nM.h) 2210 p.o. T _max (hour) 0.5 C _max (nM) 1495 AUC _0-last (nM.h) 3854 F% 89.1

Conclusion on the results of testing

The compound of the present invention exhibits good pharmacokinetic characteristics, in vivo plasma concentration and bioavailability.

Test Example 4: In Vivo Efficacy Study in Tumor Necrosis Factor (TNF-α) Triggered Systemic Inflammatory Response Syndrome (SIRS) Mouse Model

Reagent

Reagent Brand Part number or batch number Tumor necrosis factor (TNF-α) Pepro Technology 315-01A-1MG Caspase inhibitor zVAD GL Biochem (Shanghai) Ltd. GLS180522-01001

Devices and materials

1) Electronic scale: Changzhou Tianzhiping Instrument Equipment Co., Ltd. brand, model JY20002

2) Anal thermometer: brand PHYSITEMP, model BAT-12

Preparation of reagents

1) TNF-α: TNF-α was centrifuged briefly (10,000 rpm, 30s) before opening the lid. One vial of TNF-α (1 mg) was dissolved in 1 ml of sterile water to give a stock solution at a concentration of 1 mg/ml. The stock solution was stored at 4°C and can be stored for a week. 1000 μl of the stock solution was weighed and added to 5667 μl of sterile water, then 333 mg of trehalose was added and dissolved to obtain a working solution.

2) zVAD: 20 mg zVAD was weighed and added to 480 μl DMSO and the mixture was vortexed, sonicated and heated at 37°C, then 15.52 ml physiological saline was added. A working solution with a concentration of 1.25 mg/mL was obtained.

Testing stages

1) Mice were adapted to the conditions for 3 days, then randomly grouped into cages, 5 mice in each cage. Attached ear tags.

2) First, the test compound was administered to mice by gavage according to their body weight, and then zVAD was administered by intraperitoneal injection at a dosage of 250 μg per mouse. After intraperitoneal injection of zVAD, anal temperature was immediately measured (0 hour) and time was recorded. The time when mice were injected with zVAD was taken as zero hour (0 hour).

3) After 15 minutes, TNF-α was injected into the tail vein at a dosage of 200 μl per mouse, and anal temperature was measured after 15 minutes (0.5 hour).

4) After 1 hour, zVAD was administered by intraperitoneal injection at a dosage of 125 μg per mouse, and anal temperature was immediately measured (1 hour). Anal temperature was measured again after 2 hours, 3 hours and 4 hours.

Test results

The test results for the compound of the present invention are shown in Table 4.

Table 4. Body temperature and survival rate of mice Groups Number of animals in each group Decrease in body temperature in mice (°C) Mice Survival Rate at 4 Hours 1 hour 2 h 3 h 4 h TNF-α + zVAD+ vehicle 5 2.94 9.02 - - 0% after 2 hours TNF-α+zVAD+
WX009 (10mg/kg) 5 0.94 1.2 1.42 1.52 100 %

Conclusion on the results of testing

The compound of the present invention showed a very good protective effect in a mouse model of inhibition of systemic inflammatory response syndrome (SIRS) driven by TNF-α/zVAD, where the decrease in body temperature in mice was significantly reduced and the survival rate of animals increased from 0% to 100%.

Test Example 5: In Vitro Permeability Test on Caco-2 Cells

Cell culture

Cells were incubated with MEM supplemented with 2 mM L-glutamine, 10% fetal bovine serum (FBS), 100 U/ml penicillin-G, and 100 μg/ml streptomycin. Cell incubation conditions: 37±1°C, 5% CO ₂ and saturated humidity. When the cells grew to 80-90% confluence, trypsin (0.05% w/v)/EDTA (0.02% w/v) was added. Cells were seeded in a BD Falcon's Transwell 96-well plate at a density of 1×10 ⁵ cells/cm ² . Cells were incubated in a carbon dioxide incubator for 22 days and then used for transport testing.

Transport testing

Hank's balanced salt solution containing 10 mM HEPES (pH 7.40±0.05) was used in this assay as a transport testing buffer. Two-way transport of the test compound and the drug digoxin at a concentration of 2 μM was studied, each in duplicate. The transport of fenoterol and propranolol from the apical side to the basolateral side (AB) was studied. The DMSO concentration in the incubation system was kept below 1%. After adding the sample plate with cells were incubated at 37±1°C, 5% CO ₂ and saturated humidity for 120 minutes. All samples were mixed on a vortex mixer and then centrifuged at 3220 rpm and 20°C for 20 minutes. Control and test samples were diluted 1:1 (v:v) with ultrapure water and then stored at 4°C. Liquid chromatography with tandem mass spectrometry (LC/MS/MS) was used for analysis and testing.

Sample analysis

In this test, liquid chromatography with tandem mass spectrometry (LC/MS/MS) was used to semi-quantitatively analyze the ratio of peak areas of the test compound and control substances fenoterol, propranolol and digoxin relative to the internal standard in the stock solution, in the acceptor compartment solution and in the dosing supernatant. wells.

Calculation of results

The apparent permeability coefficient (P _app , cm/s), efflux coefficient and extraction coefficient were calculated using the formulas below.

The apparent permeability coefficient (P _app , cm/s) was calculated using the following formula:

P _app = (dC _r /d _t ) × V _r / (A × C ₀ )

where dC _r /d _t is the cumulative concentration of the compound on the acceptor side per unit time (µM/s); _Vr is the volume of the solution from the acceptor side (the volumes of solutions from the apical side and from the basolateral side were 0.075 ml and 0.250 ml, respectively); A is the relative surface area of the cell monolayer (0.0804 cm ² ); C ₀ is the ratio of the peak areas of the initial concentration (nM) of the test compound and control compounds from the donor side.

The efflux coefficient was calculated using the following formula:

Efflux coefficient = P _app (BA) / P _app (AB)

The release ratio was calculated using the following formula:

% extraction ratio = 100 × [(V _r × C _r ) + (V _d × C _d )] / (V _d × C ₀ ),

where C ₀ is the initial concentration (nM) of the test compound from the donor side or the peak area ratio of the control compounds; V _d is the volume from the donor side (apical side is 0.075 ml, and basolateral side is 0.250 ml); C _d and C _r are the ratio of the peak areas of the final concentrations (nM) of the test compound or control compounds from the donor side and the acceptor side, respectively.

Test results

The test results for the compound of the present invention are shown in Table 5.

Table 5. Permeability test results on Caco-2 cells Compound Apparent permeability coefficient (10 ^-6 cm/s) Efflux coefficient WX009 18.35 0.75

Conclusion on the results of testing

The compound of the present invention is a compound with high permeability and low efflux.

Test Example 6 Determination of Plasma Protein Binding Factor (PPB)

796 µl of blank human, SD rat and CD-1 mouse plasma (Plasma purchased from Bioreclamation IVT) was weighed and test compound working solution or warfarin working solution was added so that the final concentration of test compound and warfarin in the plasma sample was 2 µM. The samples were thoroughly mixed. The final concentration of DMSO in the organic phase was 0.5%. 50 µl plasma samples with test compound and warfarin were pipetted into the wells of the plate, and an appropriate volume of blank plasma or buffer was immediately added so that the final volume in each sample well was 100 µl, where the plasma:dialysis buffer volume ratio was 1: 1. Stop reagent was then added to these samples. The resulting samples were used as T ₀ samples to determine recovery and stability. Plasma samples with test compound and warfarin were placed in the top section of each dialysis cell and dialysis buffer blank was placed in the corresponding bottom section of the dialysis cell. The dialysis plate was then sealed with a breathable membrane, placed in a humidified 5% CO ₂ incubator and incubated with shaking at 100 rpm at 37° C. for 4 hours. After the end of dialysis, 50 µl of the dialysis buffer sample and the dialysis plasma sample were pipetted into a new sample plate. An appropriate volume of the appropriate blank plasma or buffer solution was added to the samples so that the final volume in each sample well was 100 μl, where the plasma:dialysis buffer volume ratio was 1:1. All samples were subjected to protein precipitation and then analyzed by LC/MS/MS. Percent unbound (% unbound), percent bound (% bound), and percent recovered (% recovered) for a test compound were calculated using the following formulas:

% unbound = 100 * F _C / T _C ,

% bound = 100 - % unbound, and

% selection = 100 * (F _C + T _C ) / T ₀ ,

where F _C is the concentration of the compound at the buffer end of the dialysis cell; T _C is the concentration of the compound at the plasma end of the dialysis cell; and T ₀ is the concentration of the compound in the plasma sample at time zero.

Test results

The test results for the compound of the present invention are shown in Table 6.

Table 6. Results of determining the coefficient of binding to plasma proteins Compound PPB_unbound (%) WX009 5.7 (human) / 29.7 (rat) / 7.0 (mouse)

Conclusion on the results of testing

The compound of the present invention has an acceptable protein binding ratio in human, rat and mouse plasma, and the percentage of this drug compound in the unbound state in human plasma is 5.7%.

Test Example 7: Determination of Pharmacokinetic Parameters in Dogs

Testing stages

Four healthy adult Beagle dogs were selected and randomly assigned to two groups. One group was administered the test compound by intravenous injection and the other group was administered the test compound via a gastric tube. Blood samples were taken, respectively, from animals from the "intravenous" group and from the "oral" group after 0.083, 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 24 hours after administration. Blood concentration was determined by LC-MS/MS. Pharmacokinetic parameters were calculated with WinNonlin™ Version 6.3 software (Pharsight, Mountain View, CA) using the non-compartmental method and the linear logarithmic trapezoid method. AUC _0-last is the area under the plasma concentration-time curve from point 0 to the time of the last detectable concentration; C _max is the peak concentration; T _max is the time to peak; T _1/2 is the half-life; CL is the hatch rate; Vd _ss is the apparent volume of distribution; po means "by mouth"; iv means intravenous injection; and F% is oral boavailability.

Test results

The results of testing the compounds of the present invention are presented in table 7.

Table 7. Pharmacokinetic characteristics of the compounds of the present invention Connections WX009 WX013 Dosage regimen iv = 3 mg/kg; po = 10 mg/kg iv = 1 mg/kg; po = 5 mg/kg iv T _1/2 (h) 5.5 4.89 Vd _ss (l/kg) 2.4 2.02 CL(ml/min/kg) 6.2 5.18 AUC _0-last (nM.h) 20762 7417 po T _max (h) 2.0 2935 C _max (nM) 284 1.50 AUC _0-last (nM.h) 23640 27744 F% 34.2% 80.1%

Conclusion on the results of testing

The compounds of the present invention have shown good pharmacokinetic parameters and high concentration in vivo in tests on dogs. In particular, the half-life of the compounds of the present invention is about 5 hours, which is relatively ideal.

Test Example 8: Kinase Selectivity Test for a Compound of the Present Invention

The aim of this study was to determine the in vitro inhibitory activity of WX009 against various kinases. The kinases examined in this assay included 413 kinases and the activity assay was provided by Eurofins Pharma Discovery Service. In this test, a method for determining the activity of kinases using ³³ P-radioactive label was used. The WX009 concentration was 1 μM and the ATP concentration was 10 μM. The results of the study are presented in Table 8.

Table 8. Results of WX009 kinase profile evaluation Inhibition coefficient Number of kinases Name of the kinase > 80% 1 RIP1 50-79% 1 LIMK2 < 50% 411 All other kinases

Conclusion of the test results: The compound of the present invention shows a highly specific kinase selectivity and has a highly active inhibitory effect almost exclusively on RIP1. Therefore, the compound of the present invention has a low risk of affecting other untargeted kinase targets.

Claims (53)

1. A compound having formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt , Where T ₁ is selected from N and CR _1t ; T ₂ is selected from N and CR _2t ; T ₃ is CR _3t ; T ₄ is CR _4t ; E ₁ is selected from C(R _1e ) ₂ and O; cycle A is selected from 1,2,4-triazolyl, where 1,2,4-triazolyl, optionally substituted with 1, 2 or 3 halogens or C _1-3 alkyls; cycle B is selected from phenyl and L is selected from a single bond, O and C _1-3 alkylene; R ₁ is selected from H and C _1-3 alkyl, where C _1-3 alkyl is optionally substituted with 1, 2 or 3 R _b substituents; R _1t , R _2t , R _3t and R _4t are each independently selected from H, F, Cl, Br, I, CN, C _1-3 alkyl, COOH and -C(=O)NH ₂ ; R _1e is each independently selected from H and C _1-3 alkyl; R ₂ is each independently selected from H, F, Cl, Br, I, CN, C _1-3 alkyl and C _1-3 alkoxy, where C _1-3 alkyl and C _1-3 alkoxy are optionally substituted with 1, 2 or 3 substituents R _f ; n is 1, 2, 3, 4 or 5; R _b and R _f are each independently selected from F, Cl, Br, I and D. 2. The compound or stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein R ₁ is selected from H and CH ₃ , where CH ₃ is optionally substituted with 1, 2, or 3 R _b substituents. 3. A compound or stereoisomer thereof, or a pharmaceutically acceptable salt according to claim 2, wherein R ₁ is selected from H, CH ₃ and CD ₃ . 4. The compound or stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein R _1t , R _2t , R _3t and R _4t are each independently selected from H, F, Cl, Br, I, CN, CH ₃ , COOH and - C(=O) _NH2 . 5. The compound or stereoisomer or pharmaceutically acceptable salt thereof according to claim 4, wherein R _1t , R _2t , R _3t and R _4t are each independently selected from H, F, Cl, Br, I, CN and CH ₃ . 6. The compound or stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein R _{1e is} each independently selected from H and CH ₃ . 7. A compound or stereoisomer thereof, or a pharmaceutically acceptable salt according to claim 1 or 6, where E ₁ is selected from CH ₂ and O. 8. The compound or stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein R ₂ is each independently selected from H, F, Cl, Br, I, CN, CH ₃ and OCH ₃ , where CH ₃ and OCH ₃ are optionally substituted with 1 , 2 or 3 R _f substituents. 9. The compound or stereoisomer or pharmaceutically acceptable salt thereof according to claim 8, wherein R ₂ is each independently selected from H, F, Cl, Br, I, CN, CH ₃ , OCH ₃ , CF ₃ and OCF ₃ . 10. A compound or stereoisomer thereof, or a pharmaceutically acceptable salt according to claim 1, wherein cycle A is selected from Where optionally substituted with 1, 2 or 3 halo or C _1-3 alkyl. 11. The compound or its stereoisomer, or pharmaceutically acceptable salt according to claim 1, where L is selected from a single bond, -CH ₂ - and -O-. 12. The compound or its stereoisomer, or pharmaceutically acceptable salt according to any one of paragraphs. 1-3, 8 and 9 where the compound is selected from , Where n, T ₁ , T ₂ and T ₃ have the meanings specified in paragraph 1; R ₁ has the meaning specified in paragraph 1, 2 or 3; R ₂ has the meaning specified in paragraph 1, 8 or 9; R is selected from H and halo; the carbon atom with "*" is a chiral carbon atom that exists in the form of an individual (R) or (S) enantiomer or a mixture enriched in one enantiomer. 13. A compound or stereoisomer thereof, or a pharmaceutically acceptable salt according to claim 12, where the compound is selected from , Where n, T ₁ , T ₂ and T ₃ have the meanings specified in paragraph 1; R ₁ has the meaning specified in paragraph 1, 2 or 3; R ₂ has the meaning specified in paragraph 1, 8 or 9; R is selected from H and halo. 14. A compound having the formula: or a stereoisomer thereof, or a pharmaceutically acceptable salt. 15. A compound or stereoisomer thereof, or a pharmaceutically acceptable salt according to claim 14, where the compound is selected from the following: . 16. The use of a compound or its stereoisomer, or a pharmaceutically acceptable salt according to any one of paragraphs. 1-15 in the manufacture of a medicament for the treatment of a disease associated with RIP-1 kinase.