HK40088676B - Nitrile derivative that acts as inhibitor of dipeptidyl peptidase 1 and use thereof - Google Patents

Description

A nitrile derivative as an inhibitor of dipeptidyl peptidase-1 and its uses

Technical Field

This invention relates to a nitrile derivative as a dipeptidyl peptidase 1 inhibitor, its stereoisomers, deuterated derivatives, cocrystals, solvates or pharmaceutically acceptable salts, and its use in the preparation of medicaments for treating dipeptidyl peptidase 1-mediated diseases.

Background Technology

Dipeptidyl peptidase 1 (DPP1), also known as cathepsin C, is a lysosomal cysteine protease capable of removing dipeptides from the amino terminus of protein substrates. DPP1 was first discovered by Gutman and Fruton in 1948 (J Biol Chem, 174, 851-858); however, the human enzyme's cDNA was first described in 1995 (FEBS Lett, 369, 326-330). DPP1 is the only member of the papain family that functions as a tetramer, consisting of four identical subunits. Each subunit comprises an N-terminal fragment, a heavy chain, and a light chain (J Biol Chem, 270, 21626-21631).

DPP1 is highly expressed in various tissues of the lung, kidney, liver, and spleen (Biol. Chem. Hoppe Seyler 373:367-373, 1992). Consistent with its activation of serine proteases in hematopoietic stem cells, DPP1 is also relatively highly expressed in neutrophils, cytotoxic lymphocytes, natural killer cells, alveolar macrophages, and mast cells. Recent data indicate that, in addition to being an important enzyme in lysosomal protein degradation, DPP1 also plays a key role in the activation of granule serine proteases in the following cellular cells: cytotoxic T lymphocytes and natural killer cells (granulases A and B; Proc. Nat. Acad. Sci 96:8627-8632, 1999), mast cells (chymotrypsin and plasmin; J Biol. Chem. 276:18551-18556, 2001), and neutrophils (cathepsin G, elastase, and protease 3; J Clin. Invest. 109:363, 371, 2002). Once activated, these proteases can degrade various extracellular matrix components, leading to tissue damage and chronic inflammation. DPP1 is considered an effective therapeutic target because it plays a central role in activating these proteases (J Clin Invest, 2002, 109, 363-271; J Immunol, 2004, 173, 7277-7281).

Therefore, cathepsin C inhibitors have the potential to treat neutrophil-dominated inflammatory diseases such as rheumatoid arthritis, chronic obstructive pulmonary disease (COPD), emphysema, asthma, multiple sclerosis, and cystic fibrosis (Curr. Topics Med. Chem. 10:708-716, 2010; Expert Opin. Ther. Patents 20:497-506, 2010). Given the role of DPP1 in activating some pro-inflammatory serine proteases, the preparation of compounds that inhibit its activity and thus inhibit the activity of downstream serine proteases has promising clinical application prospects. Currently, relevant patents have reported the synthesis of DPP1 inhibitors. WO2004/110988 relates to certain nitrile derivatives and their use as DPP1 inhibitors. WO2009/074829 relates to peptidyl nitriles and their use as DPP1 inhibitors. WO2010/128324 relates to α-aminoamide nitriles and their use as DPP1 inhibitors. WO2012/119941 relates to peptidyl nitrile compounds and their use as DPP1 inhibitors. WO2013/041497 relates to N-[1-cyano-2-(phenyl)ethyl]-2-azabicyclo[2.2.1]heptane-3-carboxamide and its use as a DPP1 inhibitor. WO2001/096285 and WO2003/048123 relate to β-aminoamide nitriles with inhibitory activity against cysteine proteases. However, to date, there are still no marketed DPP1 inhibitors, therefore, a DPP1 inhibitor with high inhibitory activity and low toxicity remains an unmet clinical need.

Summary of the Invention

This invention first provides a compound of formula (I), (II), (III), or (IV) with high activity, high bioavailability, good pharmacokinetics, and low toxicity, including its stereoisomers, deuterated derivatives, eutectics, solvates, or pharmaceutically acceptable salts:

Wherein, ring G is a 5-12 membered carbon ring, a 5-12 membered monocyclic heterocycle containing 1-3 heteroatoms selected from N, S, and O, or a fused ring of formula (I-1), and L ₁ replaces any hydrogen atom on the ring atom of ring G (e.g., carbon atom on the 5-12 membered carbon ring, cyclic carbon atom or cyclic heteroatom on the 5-12 membered monocyclic heterocycle, or cyclic carbon atom or cyclic heteroatom on the fused ring of formula (I-1)). The carbon ring or monocyclic heterocycle is optionally replaced by 1-3 R _Gs .

Optionally, equation (I-1) has a structure

Optionally, equation (I-1) has a structure

In some embodiments, ring G is a 5-9 member monocyclic carbon ring, a 5-7 member monocyclic heterocycle containing 1-3 heteroatoms selected from N, S, and O, or a fused ring of formula (I-1). Optionally, formula (I-1) has structure (I-11), and optionally, formula (I-1) has structure (I-12).

In some embodiments, ring G is cyclopentane, cyclohexane, cycloheptane, benzene ring, or ring G optionally substituted with 1-3 R _Gs .

Optionally, equation (I-2) has a structure

Optionally, equation (I-2) has a structure

In some implementations, one of the following structures is selected:

In some embodiments, ring G is a benzene ring optionally substituted with 1-2 _RGs or

Each _RG is independently selected from deuterium, _SF5 , =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C1-4 alkoxy, _C3-6 cycloalkyloxy, C3-6 cycloalkyl, _C2-6 alkenyl, _C2-6 alkynyl, _-NHC1-4 alkyl, -N( _C1-4 alkyl) ₂ , _-COC1-4 alkyl _, -COOC1-4 alkyl, _-CONH2 , _-CONHC1-4 alkyl, _-CONHC3-6 cycloalkyl, -CON( _C1-4 alkyl) ₂ , _-NHCOC1-4 alkyl, -NHCOC3-6 _cycloalkyl , -P(O)( _{C1-4 alkyl} ) ₂ , -S(O _{)C1-4 alkyl, -S(O)2C1-4 alkyl ,} -S(O) _2C The RG may be further substituted with 1-3 groups selected from N, S, and O heteroatoms, including _3-6 cycloalkyl, -S(O)NH ₂ , -S(O)NHC _1-4 alkyl, -S(O)N(C _1-4 alkyl) _2, -S(O) ₂ NH ₂ , and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O heteroatoms. The _RG may also be further substituted with 1-3 groups selected from deuterium, C _1-4 alkyl, halo-C _1-4 alkyl, hydroxy-C _1-4 alkyl, -CONH ₂ , NH ₂ , C _1-6 alkoxy, hydroxyl, -COOH, halogen, and 5-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O heteroatoms. Further, the 5-7 membered heterocycle may be substituted with 1-2 =O, halogen, cyano, C _1-4 alkyl, or halo-C _1-4 alkyl groups.

In some embodiments, each _RG is independently selected from deuterium, _SF5 , =O, halogen, cyano, hydroxyl, _NH2 , _C1-4 alkyl, _C1-4 alkoxy, _C3-6 cycloalkyl, _C2-6 alkenyl, _C2-6 alkynyl, _-NHC1-4 alkyl, -N( _C1-4 alkyl) ₂ , _-COC1-4 alkyl, -CONH2, _-CONHC1-4 alkyl, -CON( _C1-4 alkyl) ₂ , _-NHCOC1-4 alkyl, -P(O)( _C1-4 alkyl) ₂ , and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O _, wherein the _RG is optionally further composed of 1-3 heteroatoms selected from deuterium, _C1-4 alkyl, halogenated _C1-4 alkyl, hydroxyl _C1-4 alkyl, _-CONH2 , _NH2 , C _{The 1-6} alkoxy, hydroxyl, halogen, and groups containing 1-3 5-7 membered heterocycles selected from N, S, and O heteroatoms are substituted; further, the 5-7 membered heterocycles are optionally substituted by 1-2 =O, halogen, cyano, _C1-4 alkyl, or halo _-C1-4 alkyl groups.

In some embodiments, each _RG is independently selected from F, Cl, Br, I, methyl, ethyl, propyl, _SF5 , and CN; the methyl, ethyl, and propyl groups may optionally be further substituted with 1-3 groups selected from deuterium, F, Cl, Br, and I;

Cy is a 5-12 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the heterocycle is optionally surrounded by 1-3 heteroatoms selected from deuterium, =O, halogen, cyano, hydroxyl, _NH₂ , -COOH, _C1-4 alkyl, halo- _C1-4 alkyl, hydroxy- _C1-4 alkyl, _C1-4 alkoxy, _C3-6 cycloalkyloxy, _C1-6 alkoxyalkyl, halo- _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, -NHC1-4 alkyl, -N( _C1-4 alkyl) _₂ , _-COC1-4 alkyl, _-COOC1-4 alkyl, _-CONH₂ , _{-CONHC1-4 alkyl} , -CON( _C1-4 alkyl) _₂ , _-NHCOC1-4 alkyl, -S(O) _C1-4 alkyl, -S(O) _₂C Substitution of _1-4 alkyl, -S(O)NH ₂ , -S(O)NHC _1-4 alkyl, -S(O)N(C _1-4 alkyl) ₂ , -S(O) _{2NH 2 ,} -S(O) _{2NHC 1-4} alkyl, -S(O) _2N (C _1-4 alkyl) ₂ groups;

In some specific embodiments, Cy is a 5-12 membered monocyclic heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the monocyclic heterocycle is optionally surrounded by 1-3 heteroatoms selected from deuterium, =O, halogen, cyano, hydroxyl, _NH₂ , -COOH, _C1-4 alkyl, halo- _C1-4 alkyl, hydroxy- _C1-4 alkyl, _C1-4 alkoxy, _C3-6 cycloalkoxy, _C1-6 alkoxyalkyl, halo- _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, _-NHC1-4 alkyl, -N( _C1-4 alkyl) _₂ , _-COC1-4 alkyl, _-COOC1-4 alkyl, _-CONH₂ , -CONHC1-4 alkyl, -CON( _{C1-4 alkyl} ) _₂ , _-NHCOC1-4 alkyl, -S(O) _C1-4 alkyl, -S(O) _₂C Substitution of _1-4 alkyl, -S(O)NH ₂ , -S(O)NHC _1-4 alkyl, -S(O)N(C _1-4 alkyl) ₂ , -S(O) _{2NH 2 ,} -S(O) _{2NHC 1-4} alkyl, -S(O) _2N (C _1-4 alkyl) ₂ groups;

In some specific embodiments, Cy is a heterocycle containing 1-3 heteroatoms selected from N, S, and O, comprising: 5-8 membered monocyclic heterocycles, 7-10 membered spirocyclic heterocycles, 6-9 membered bridged heterocycles, or 6-10 membered fused heterocycles. Cy is optionally surrounded by 1-3 heteroatoms selected from =O, halogen, cyano, hydroxyl, _NH₂ , -COOH, C₁ _-4 alkyl, halo-C₁ _-4 alkyl, hydroxy-C₁ _-4 alkyl, C₁ _-4 alkoxy, C₃ _-6 cycloalkoxy, C₁ _-6 alkoxyalkyl, halo-C₁ _-4 alkoxy, -NHC₁ _-4 alkyl, -N(C₁ _-4 alkyl) _₂ , -COC₁ _-4 alkyl, -CONH₂, -CONHC₁ _{-4 alkyl} , -CON(C₁ _-4 alkyl) _₂ , -NHCOC₁ _-4 alkyl, -S(O)C₁ _-4 alkyl, -S(O) _₂C₂ Substitution of _1-4 alkyl, -S(O)NH ₂ , -S(O)NHC _1-4 alkyl, -S(O)N(C _1-4 alkyl) ₂ , -S(O) _{2NH 2 ,} -S(O) _{2NHC 1-4} alkyl, -S(O) _2N (C _1-4 alkyl) ₂ groups;

In some specific embodiments, Cy is a 5-8 membered monocyclic heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the monocyclic heterocycle is optionally surrounded by 1-3 heteroatoms selected from =O, halogen, cyano, hydroxyl, _NH₂ , -COOH, _C1-4 alkyl, halo- _C1-4 alkyl, hydroxy- _C1-4 alkyl, _C1-4 alkoxy, _C3-6 cycloalkoxy, _C1-6 alkoxyalkyl, halo- _C1-4 alkoxy, _-NHC1-4 alkyl, -N( _C1-4 alkyl) _₂ , _-COC1-4 alkyl, _-CONH₂ , _-CONHC1-4 alkyl, -CON( _C1-4 alkyl) _₂ , -NHCOC1-4 alkyl, -S(O) _{C1-4 alkyl} , -S(O) _{₂C1-4 alkyl} , -S(O) _NH₂ , -S(O)NHC Substitution of _1-4 alkyl, -S(O)N(C _1-4 alkyl) ₂ , -S(O) _{2NH 2 ,} -S(O) _{2NHC 1-4} alkyl, -S(O) _2N (C _1-4 alkyl) ₂ groups;

In some specific embodiments, Cy is a 5-8 membered monocyclic heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the monocyclic heterocycle is optionally substituted by 1-3 groups selected from =O, halogen, cyano, hydroxyl, _NH₂ , -COOH, C₁ _-4 alkyl, halo-C₁ _-4 alkyl, C₁ _-4 alkoxy, C₃ _-6 cycloalkoxy, halo-C₁ _- 4 alkoxy, -NHC₁ _-4 alkyl, -N(C₁ _-4 alkyl) _₂ , _-CONH₂ , -CONHC₁- ₄ alkyl, -CON(C₁ _{-4 alkyl)₂ ,} -S(O) _₂NH₂ , -S(O)₂NHC₁ _{- 4} alkyl, -S(O) _₂N (C₁ _-4 alkyl) _₂ .

In some specific implementation schemes, Cy is

Wherein Rc is H, =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, halo- _C1-4 alkyl, hydroxy _-C1-4 alkyl, _C1-4 alkoxy, _C1-6 alkoxyalkyl, or halo- _C1-4 alkoxy;

In some specific embodiments, Rc is H, halogen, _C1-2 alkyl, or _C1-2 alkoxy;

W is C(=O), C(=S), C(= _NRW ), S(=O) or S(=O) ₂ , and _RW is OH, CN, or _C1-4 alkyl.

In some specific implementations, W is C (＝O);

M is NR _M or O, and _RM is H, _C1-4 alkyl or _C3-6 cycloalkyl;

In some specific implementation schemes, M is NH;

_R1 , _R2 , and _R3 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 ynyl, _C3-6 cycloalkyl, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O. The alkyl, alkoxy, alkenyl, ynyl, cycloalkyl, and heterocycles are optionally substituted with 1-3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 , and -COOH. Optionally, _R1 and _R2 form a _C3-6 cycloalkyl or a 4-7 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O with the carbon atoms they are attached to. The cycloalkyl or heterocycle is optionally substituted with 1-3 groups selected from =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C2-6 alkenyl, C2-6 ynyl, and _C3-6 cycloalkyl. Substitution of _3-6 cycloalkyl groups;

In some specific embodiments, _R1 , _R2 and _R3 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl and _C2-6 alkynyl, wherein the alkyl, alkoxy, alkenyl and alkynyl groups are optionally substituted by 1 to 3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 and COOH;

Optionally, _R1 and _R2 form a _C3-6 cycloalkyl group or a 4-7 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the cycloalkyl group or heterocycle is optionally substituted by 1-3 groups selected from halogen, cyano, hydroxyl, _NH2 , COOH, and _C1-4 alkyl groups.

In some specific embodiments, _R1 , _R2 and _R3 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, wherein the alkyl or alkoxy group is optionally substituted by 1 to 3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 and COOH;

Optionally, _R1 and _R2 form a _C3-6 cycloalkyl group with the carbon atoms they are attached to, wherein the cycloalkyl group is optionally substituted by 1-3 groups selected from halogen, cyano, hydroxy, _NH2 , COOH, and _C1-4 alkyl groups;

In some specific embodiments, _R1 , _R2 and _R3 are each independently selected from H, deuterium, F, Cl, Br, methyl, ethyl, methoxy or ethoxy, wherein the methyl, ethyl, methoxy or ethoxy group is optionally substituted by 1 to 3 groups selected from F, Cl, Br, cyano, hydroxyl and _NH2 ;

_L1 is a bond, _C1-3 alkylene, -NH-, -N( _C1-4 alkyl)-, -O-, -S-, _C2-6 alkenyl, _C2-6 alynyl, -CO-, or -CONH-, wherein the alkylene, alkenyl, or alynyl group is optionally substituted by 1-3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 , and -COOH;

In some specific embodiments, L1 is a bond, _C1-3 alkylene, -NH-, -O-, -S-, _C2-6 alkenyl, _C2-6 alynyl, or -CO-, wherein the alkylene, alkenyl, or alynyl group is optionally substituted by 1-3 groups selected from halogen, cyano, hydroxyl, and _NH2 ;

In some specific implementations, L1 is the key,

_Y1 and _Y2 are each independently selected from _CR4 or N;

In some specific implementation schemes, _Y1 and _Y2 are CH;

Each R ₄ is independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, _C3-6 cycloalkyl, cyano, hydroxyl, NH ₂ , -NHC _1-4 alkyl, -N(C _1-4 alkyl) ₂ , -COOH, -COC _1-4 alkyl, -COOC _1-4 alkyl, -CONHC _1-4 alkyl, -CON(C _1-4 alkyl) ₂ , -NHCOC _1-4 alkyl, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O; the alkyl, alkenyl, alkynyl, cycloalkyl, and heterocyclic groups in R ₄ are optionally substituted with 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, NH ₂ , and -COOH;

In some specific embodiments, each _R4 is independently selected from H, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, _C3-6 cycloalkyl, cyano, hydroxyl, _NH2 ; the alkyl, alkenyl, alkynyl, and cycloalkyl groups in _R4 are optionally substituted by 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, _NH2 , and COOH;

In some specific implementation schemes, _R4 is H;

_X1 , _X2 , _X3 and _X4 are each independently selected from bond, _NR5 , O, _CR6R7 , S _, S(O) and S(O) ₂ , and at most one of _X1 , _X2 , _X3 and _X4 is a bond;

Each R ₅ is independently selected from H, C _1-4 alkyl, -COC _1-4 alkyl, C _2-6 alkenyl, C _2-6 ynyl and C _3-6 cycloalkyl; the alkyl, alkenyl, ynyl and cycloalkyl are optionally substituted by 1 to 3 groups selected from deuterium, halogen, cyano, hydroxyl, NH ₂ and COOH;

In some specific embodiments, each R ₅ is independently selected from H, C _1-4 alkyl, -COC _1-4 alkyl, C _3-6 cycloalkyl; the alkyl or cycloalkyl group is optionally substituted with 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, NH ₂ and COOH;

In some specific implementations, _R5 is methyl;

_R6 and _R7 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, _C3-6 cycloalkyl, cyano, hydroxyl, _NH2 , -COOH, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, and heterocycles are optionally substituted by 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, _NH2 , and -COOH;

Alternatively, _R6 and _R7 form = 0;

Optionally, two _R5s on adjacent ring atoms, two _R6s on adjacent ring atoms, or _R5s and _R6s on adjacent ring atoms together with the atoms they are connected to form double bonds in _X1 , _X2 , _X3 and _X4 ;

Optionally, _R6 and _R7 on the same carbon atom form a _C3-12 carbon ring with the carbon atom to which they are attached, or a 4-7 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O. The carbon ring or heterocycle is optionally substituted by 1-3 groups selected from =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 ynyl, and _C3-6 cycloalkyl.

In some specific embodiments, _R6 and _R7 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, and _C3-6 cycloalkyl; the alkyl, alkoxy, alkenyl, alkynyl, and cycloalkyl groups are optionally substituted by 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, _NH2 , and COOH;

Alternatively, _R6 and _R7 form = 0;

Optionally, two _R5s on adjacent ring atoms, two _R6s on adjacent ring atoms, or _R5s and _R6s on adjacent ring atoms together with the atoms they are connected to form double bonds in _X1 , _X2 , _X3 and _X4 ;

Optionally, _R6 and _R7 on the same carbon atom form a _C3-6 cycloalkyl group or a 4-7 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the cycloalkyl group or heterocycle is optionally substituted by 1-3 groups selected from =O, halogen, cyano, hydroxyl, _NH2 , COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 ynyl and _C3-6 cycloalkyl groups;

In some specific embodiments, _R6 and _R7 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, and _C1-4 alkoxy; the alkyl and alkoxy groups are optionally substituted by 1 to 3 groups selected from deuterium, halogen, cyano, hydroxyl, _NH2 , and COOH;

Alternatively, _R6 and _R7 form = 0;

Optionally, two _R5s on adjacent ring atoms, two _R6s on adjacent ring atoms, or _R5s and _R6s on adjacent ring atoms together with the atoms they are connected to form double bonds in _X1 , _X2 , _X3 and _X4 ;

Z is CH or N;

The compounds of formulas (I)-(III) must satisfy one of the conditions (1)-(4).

(1) A is a six-membered heteroaryl or a 5-10-membered non-aromatic heterocycle, wherein the heteroaryl or non-aromatic heterocycle contains 1-3 heteroatoms selected from N, S, and O, and the hetero A is optionally substituted by 1-3 R ₈ atoms;

* indicates the connection end with an alkyl carbon;

Ring B is a _C3-12 carbon ring or contains 1-3 4-7 membered heterocycles selected from N, S, and O heteroatoms;

Each R ₈ is independently selected from H, =O, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, -SC _1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, C3-6 cycloalkyl, cyano, hydroxyl, NH ₂ , -NHC _1-4 alkyl, -N( _{C1-4 alkyl} ) ₂ , -COOH, -COC _1-4 alkyl, -COOC _1-4 alkyl, -CONHC _1-4 alkyl, -CON( _C1-4 alkyl) ₂ , -NHCOC _1-4 alkyl, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O; the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, and heterocyclic groups in R ₈ are optionally substituted with 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, NH ₂ , and -COOH;

And Cy is neither substituted nor unsubstituted.

In some specific embodiments, A is a six-membered heteroaryl or a 5-7-membered non-aromatic monocyclic heterocycle, wherein the heteroaryl or non-aromatic monocyclic heterocycle contains 1-3 heteroatoms selected from N, S, and O, and A is optionally substituted by 1-3 R ₈ atoms;

* indicates the connection end with an alkyl carbon;

Ring B is a _C4-6 carbon ring or contains 1-3 5-6 membered heterocycles selected from N, S, and O heteroatoms;

Each R ₈ is independently selected from H, =O, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _-SC1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, cyano, hydroxyl, -COOH, NH2, and _C3-6 cycloalkyl; said alkyl, alkoxy, alkenyl, alkynyl, and cycloalkyl are optionally substituted by 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, _{NH2 ,} and -COOH;

In some other specific implementations, A is ring A, which is optionally replaced by 1-3 R ₈ ;

Each _R8 is independently selected from halogen, =O, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 ynyl and _C3-6 cycloalkyl;

* indicates the connection end with an alkyl carbon;

r is an integer from 1 to 3;

E is selected from NH, S, and O;

In other specific embodiments, A is a ring A optionally substituted with 1-3 R _8s ; the substituents are located at any substituted position on the ring A, including on the carbon atom of the benzene ring and on the carbon atom or heteroatom of the ring fused with it;

Each _R8 is independently selected from halogen, =O, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C1-4 alkoxy, _-SC1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl and _C3-6 cycloalkyl;

* indicates the connection end with an alkyl carbon;

or,

(2) A is a five-membered heteroaryl group containing 1-3 heteroatoms selected from N, S, and O, wherein A is optionally substituted by 1-3 R ₈ atoms;

Each R ₈ is independently selected from H, =O, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, -SC _1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, _C3-6 cycloalkyl, cyano, hydroxyl, NH ₂ , -NHC _1-4 alkyl, -N( _C1-4 alkyl) ₂ , -COOH, -COC _1-4 alkyl, -COOC _1-4 alkyl, -CONHC _1-4 alkyl, -CON( _C1-4 alkyl) ₂ , -NHCOC _1-4 alkyl, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O; the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, and heterocyclic groups in R ₈ are optionally substituted with 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, NH ₂ , and -COOH;

And Cy is neither substituted nor unsubstituted.

In some specific embodiments, A is a five-membered heteroaryl group containing 1-3 heteroatoms selected from N, S, and O, wherein A is optionally substituted by 1-3 R ₈ atoms;

Each R ₈ is independently selected from H, =O, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _-SC1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, cyano, hydroxyl, -COOH, NH2, and _C3-6 cycloalkyl; said alkyl, alkoxy, alkenyl, alkynyl, and cycloalkyl are optionally substituted by 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, _{NH2 ,} and -COOH;

And Cy is neither substituted nor unsubstituted.

In some specific implementations, A is optionally replaced by 1-3 R ₈ ;

Each _R8 is independently selected from halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl and _C3-6 cycloalkyl;

_Z1 and _Z2 are each independently CH or N;

_Z3 is S, O, or NH;

* indicates the connection end with an alkyl carbon;

In some specific embodiments, A is optionally substituted with 1-3 groups selected from halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl and _C3-6 cycloalkyl groups;

And Cy is

* indicates the connection end with an alkyl carbon;

or,

(3) A is * to indicate the connection end with the alkyl carbon;

_RA1 , _RA2 , _RA3 , and _RA4 are each independently _R8 , and _RA1 , _RA2 , _RA3 , and _RA4 are not simultaneously H;

Each R ₈ is independently selected from H, =O, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, -SC _1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, _C3-6 cycloalkyl, cyano, hydroxyl, NH ₂ , -NHC _1-4 alkyl, -N( _C1-4 alkyl) ₂ , -COOH, -COC _1-4 alkyl, -COOC _1-4 alkyl, -CONHC _1-4 alkyl, -CON( _C1-4 alkyl) ₂ , -NHCOC _1-4 alkyl, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O; the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, and heterocyclic groups in R ₈ are optionally substituted with 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, NH ₂ , and -COOH;

Cy is a 5-12 membered monocyclic heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the monocyclic heterocycle is optionally surrounded by 1-3 heteroatoms selected from deuterium, =O, halogen, cyano, hydroxyl, _NH₂ , -COOH, _C1-4 alkyl, halo _{-C1-4 alkyl} , hydroxy- _C1-4 alkyl, _C1-4 alkoxy, C3-6 cycloalkoxy, _C1-6 alkoxyalkyl, halo- _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, -NHC1-4 alkyl, -N _{(C1-4 alkyl} ) _₂ , _-COC1-4 alkyl, _-COOC1-4 alkyl, _-CONH₂ , _-CONHC1-4 alkyl, -CON( _C1-4 alkyl) _₂ , _-NHCOC1-4 alkyl, -S(O) _C1-4 alkyl, -S(O) _₂C Substitution of _1-4 alkyl, -S(O)NH ₂ , -S(O)NHC _1-4 alkyl, -S(O)N(C _1-4 alkyl) ₂ , -S(O) _{2NH 2 ,} -S(O) _{2NHC 1-4} alkyl, -S(O) _2N (C _1-4 alkyl) ₂ groups;

And Cy is neither substituted nor unsubstituted.

In some specific implementations, A is * to indicate the connection end with the alkyl carbon;

_RA1 , _RA2 , _RA3 , and _RA4 are each independently _R8 , and _RA1 , _RA2 , _RA3 , and _RA4 are not simultaneously H;

Each R ₈ is independently selected from H, =O, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _-SC1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, cyano, hydroxyl, -COOH, NH2, and _C3-6 cycloalkyl; said alkyl, alkoxy, alkenyl, alkynyl, and cycloalkyl are optionally substituted by 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, _{NH2 ,} and -COOH;

Cy is a 5-8 membered monocyclic heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the monocyclic heterocycle is optionally surrounded by 1-3 heteroatoms selected from =O, halogen, cyano, hydroxyl, _NH₂ , -COOH, _C1-4 alkyl, halo _{-C1-4 alkyl} , hydroxy- _C1-4 alkyl, _C1-4 alkoxy, C3-6 cycloalkoxy, _C1-6 alkoxyalkyl, halo- _C1-4 alkoxy, _-NHC1-4 alkyl, -N( _C1-4 alkyl) _₂ , _-COC1-4 alkyl, -CONH₂, _-CONHC1-4 alkyl, -CON( _{C1-4 alkyl} ) _₂ , _-NHCOC1-4 alkyl, -S ₍ O) _C1-4 alkyl, -S(O) _₂C1-4 alkyl, -S(O) _NH₂ , -S(O)NHC Substitution of _1-4 alkyl, -S(O)N(C _1-4 alkyl) ₂ , -S(O) _{2NH 2 ,} -S(O) _{2NHC 1-4} alkyl, -S(O) _2N (C _1-4 alkyl) ₂ groups;

In some specific implementations, A is * to indicate the connection end with the alkyl carbon;

_RA1 , _RA2 , _RA3 , and _RA4 are each independently _R8 , and _RA1 , _RA2 , _RA3 , and _RA4 are not simultaneously H;

Each R ₈ is independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, cyano, hydroxyl, _NH2 , COOH; the alkyl and alkoxy groups are optionally selected from 1 to 3 of deuterium, halogen, cyano, hydroxyl, _NH2 , and COOH;

And Cy is

or,

(4) A is * to indicate that it is connected to an alkyl carbon;

_R1 , _R2 , and _R3 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, _C3-6 cycloalkyl, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O. The alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, and heterocycles are optionally substituted by 1-3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 , and -COOH, and _R1 , _R2 , and _R3 are not all H.

Optionally, _R1 and _R2 form a _C3-6 cycloalkyl group or a 4-7 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the cycloalkyl group or heterocycle is optionally substituted with 1-3 groups selected from =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 ynyl, and _C3-6 cycloalkyl.

In some specific implementations, A is * indicating a connection to an alkyl carbon;

_R1 , _R2 , and _R3 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, and _C2-6 alkynyl, wherein the alkyl, alkoxy, alkenyl, and alkynyl groups are optionally substituted by 1 to 3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 , and COOH, and _R1 , _R2 , and _R3 are not all H;

Optionally, _R1 and _R2 form a _C3-6 cycloalkyl group with the carbon atoms they are attached to, wherein the cycloalkyl group is optionally substituted with 1-3 groups selected from =O, halogen, cyano, hydroxyl, _NH2 , COOH, _C1-4 alkyl, _C2-6 alkenyl, and _C2-6 ynyl.

In some specific implementations, A is * indicating a connection to an alkyl carbon;

_R1 , _R2 , and _R3 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, and _C2-6 alkynyl, wherein the alkyl, alkoxy, alkenyl, and alkynyl groups are optionally substituted by 1 to 3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 , and COOH, and _R1 , _R2 , and _R3 are not all H;

Optionally, _R1 and _R2 form a _C3-6 cycloalkyl group with the carbon atoms they are attached to, wherein the cycloalkyl group is optionally substituted by 1-3 groups selected from halogen, cyano, hydroxyl, _NH2 , COOH;

In this invention, the term "connection site" is used.

As a more specific first technical solution of the present invention, the present invention provides a compound of formula (I), including its stereoisomer, deuterated derivative, eutectic, solvate, or pharmaceutically acceptable salt:

Wherein, ring G is a 5-12 membered carbon ring, a 5-12 membered monocyclic heterocycle containing 1-3 heteroatoms selected from N, S, and O, or a fused ring of formula (I-1), and L ₁ replaces any hydrogen atom on the ring atom of ring G (e.g., carbon atom on the 5-12 membered carbon ring, cyclic carbon atom or cyclic heteroatom on the 5-12 membered monocyclic heterocycle, or cyclic carbon atom or cyclic heteroatom on the fused ring of formula (I-1)). The carbon ring or monocyclic heterocycle is optionally replaced by 1-3 R _Gs .

Optionally, equation (I-1) has a structure

Optionally, equation (I-1) has a structure

Each _RG is independently selected from deuterium, _SF5 , =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, halo- _C1-4 alkyl, _C1-4 alkoxy, _C3-6 cycloalkyloxy, _C3-6 cycloalkyl, _C2-6 alkenyl, _C2-6 alkynyl, _-NHC1-4 alkyl, -N( _C1-4 alkyl) ₂ , _-COC1-4 alkyl, _-COOC1-4 alkyl, _-CONH2 , _-CONHC1-4 alkyl, -CONHC3-6 cycloalkyl, -CON( _C1-4 alkyl) _{2 , -NHCOC1-4} alkyl, _-NHCOC3-6 cycloalkyl, -P(O)( _C1-4 alkyl) ₂ , -S( _O ) _C1-4 alkyl, -S(O) _2C1-4 alkyl, -S(O) _2C The RG may be further substituted with 1-3 groups selected from N, S, and O heteroatoms, including _3-6 cycloalkyl, -S(O)NH ₂ , -S(O)NHC _1-4 alkyl, -S(O)N(C _1-4 alkyl) _2, -S(O) ₂ NH ₂ , and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O heteroatoms. The _RG may also be further substituted with 1-3 groups selected from deuterium, C _1-4 alkyl, halo-C _1-4 alkyl, hydroxy-C _1-4 alkyl, -CONH ₂ , NH ₂ , C _1-6 alkoxy, hydroxyl, -COOH, halogen, and 5-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O heteroatoms. Further, the 5-7 membered heterocycle may be substituted with 1-2 groups selected from =O, halogen, cyano, C _1-4 alkyl, and halo-C _1-4 alkyl.

Cy is a 5-12 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the heterocycle is optionally surrounded by 1-3 heteroatoms selected from deuterium, =O, halogen, cyano, hydroxyl, _NH₂ , -COOH, _C1-4 alkyl, halo- _C1-4 alkyl, hydroxy- _C1-4 alkyl, _C1-4 alkoxy, _C3-6 cycloalkyloxy, _C1-6 alkoxyalkyl, halo- _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, -NHC1-4 alkyl, -N( _C1-4 alkyl) _₂ , _-COC1-4 alkyl, _-COOC1-4 alkyl, _-CONH₂ , _{-CONHC1-4 alkyl} , -CON( _C1-4 alkyl) _₂ , _-NHCOC1-4 alkyl, -S(O) _C1-4 alkyl, -S(O) _₂C Substitution of _1-4 alkyl, -S(O)NH ₂ , -S(O)NHC _1-4 alkyl, -S(O)N(C _1-4 alkyl) ₂ , -S(O) _{2NH 2 ,} -S(O) _{2NHC 1-4} alkyl, and -S(O) _2N (C _1-4 alkyl) ₂ groups;

W is C(=O), C(=S), C(=NR _W ), S(=O) or S(=O) ₂ ;

M is NR _M or O;

R _W is OH, CN, or _C1-4 alkyl;

_RM is H, _C1-4 alkyl, or _C3-6 cycloalkyl;

_R1 , _R2 , and _R3 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, _C3-6 cycloalkyl, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O. The alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, and heterocycles are optionally substituted with 1-3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 , and -COOH.

Optionally, _R1 and _R2 form a _C3-6 cycloalkyl group or a 4-7 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the cycloalkyl group or heterocycle is optionally substituted with 1-3 groups selected from =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 ynyl, and _C3-6 cycloalkyl.

_L1 is a bond, _C1-3 alkylene, -NH-, -N( _C1-4 alkyl)-, -O-, -S-, _C2-6 alkenyl, _C2-6 alynyl, -CO-, or -CONH-, wherein the alkylene, alkenyl, or alynyl group is optionally substituted by 1-3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 , and -COOH;

_Y1 and _Y2 are each independently selected from _CR4 and N;

Each R ₄ is independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, _C3-6 cycloalkyl, cyano, hydroxyl, NH ₂ , -NHC _1-4 alkyl, -N(C _1-4 alkyl) ₂ , -COOH, -COC _1-4 alkyl, -COOC _1-4 alkyl, -CONHC _1-4 alkyl, -CON(C _1-4 alkyl) ₂ , -NHCOC _1-4 alkyl, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O; the alkyl, alkenyl, alkynyl, cycloalkyl, and heterocyclic groups in R ₄ are optionally substituted with 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, NH ₂ , and -COOH;

_X1 , _X2 , _X3 and _X4 are each independently selected from bond, _NR5 , O, _CR6R7 , S _, S(O) and S(O) ₂ , and at most one of _X1 , _X2 , _X3 and _X4 is a bond;

Each R ₅ is independently selected from H, C _1-4 alkyl, -COC _1-4 alkyl, C _2-6 alkenyl, C _2-6 ynyl and C _3-6 cycloalkyl, wherein the alkyl, alkenyl, ynyl and cycloalkyl are optionally substituted by 1 to 3 groups selected from deuterium, halogen, cyano, hydroxyl, NH ₂ and COOH;

_R6 and _R7 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, _C3-6 cycloalkyl, cyano, hydroxyl, _NH2 , -COOH, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O; wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, and heterocycles are optionally substituted with 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, _NH2 , and -COOH;

Alternatively, _R6 and _R7 form = 0;

Optionally, two _R5s on adjacent ring atoms, two _R6s on adjacent ring atoms, or _R5s and _R6s on adjacent ring atoms together with the atoms they are connected to form double bonds in _X1 , _X2 , _X3 and _X4 ;

Optionally, _R6 and _R7 on the same carbon atom form a _C3-12 carbon ring with the carbon atom to which they are attached, or a 4-7 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O. The carbon ring or heterocycle is optionally substituted by 1-3 groups selected from =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 ynyl, and _C3-6 cycloalkyl.

The condition is that,

(1) A is a six-membered heteroaryl or a 5-10-membered non-aromatic heterocycle, wherein A is optionally substituted by 1-3 R ₈ ;

* indicates the connection end with an alkyl carbon;

Ring B is a _C3-12 carbon ring or a 4-7 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the carbon ring or heterocycle is optionally substituted by 1-3 groups selected from =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C1-4 alkoxy, _-SC1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, and _C3-6 cycloalkyl.

Each R ₈ is independently selected from H, =O, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, -SC _1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, C3-6 cycloalkyl, cyano, hydroxyl, NH ₂ , -NHC _1-4 alkyl, -N( _{C1-4 alkyl} ) ₂ , -COOH, -COC _1-4 alkyl, -COOC _1-4 alkyl, -CONHC _1-4 alkyl, -CON( _C1-4 alkyl) ₂ , -NHCOC _1-4 alkyl, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O; the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, and heterocyclic groups in R ₈ are optionally substituted with 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, NH ₂ , and -COOH;

And Cy is neither substituted nor unsubstituted.

or,

(2) A is a five-membered heteroaryl group containing 1-3 heteroatoms selected from N, S, and O, wherein A is optionally substituted by 1-3 heteroatoms selected from R ₈ ;

Each R ₈ is independently selected from H, =O, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, -SC _1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, _C3-6 cycloalkyl, cyano, hydroxyl, NH ₂ , -NHC _1-4 alkyl, -N( _C1-4 alkyl) ₂ , -COOH, -COC _1-4 alkyl, -COOC _1-4 alkyl, -CONHC _1-4 alkyl, -CON( _C1-4 alkyl) ₂ , -NHCOC _1-4 alkyl, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O; the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, and heterocyclic groups in R ₈ are optionally substituted with 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, NH ₂ , and -COOH;

And Cy is neither substituted nor unsubstituted.

or,

(3) A is * to indicate the connection end with the alkyl carbon;

_RA1 , _RA2 , _RA3 , and _RA4 are each independently _R8 , and _RA1 , _RA2 , _RA3 , and _RA4 are not simultaneously H;

Each R ₈ is independently selected from H, =O, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, -SC _1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, _C3-6 cycloalkyl, cyano, hydroxyl, NH ₂ , -NHC _1-4 alkyl, -N( _C1-4 alkyl) ₂ , -COOH, -COC _1-4 alkyl, -COOC _1-4 alkyl, -CONHC _1-4 alkyl, -CON( _C1-4 alkyl) ₂ , -NHCOC _1-4 alkyl, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O; the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, and heterocyclic groups in R ₈ are optionally substituted with 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, NH ₂ , and -COOH;

Cy is a 5-12 membered monocyclic heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the monocyclic heterocycle is optionally surrounded by 1-3 heteroatoms selected from deuterium, =O, halogen, cyano, hydroxyl, _NH₂ , -COOH, _C1-4 alkyl, halo _{-C1-4 alkyl} , hydroxy- _C1-4 alkyl, _C1-4 alkoxy, C3-6 cycloalkyloxy, _C1-6 alkoxyalkyl, halo- _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, -NHC1-4 alkyl, -N _{(C1-4 alkyl} ) _₂ , _-COC1-4 alkyl, _-COOC1-4 alkyl, _-CONH₂ , _-CONHC1-4 alkyl, -CON( _C1-4 alkyl) _₂ , _-NHCOC1-4 alkyl, -S(O) _C1-4 alkyl, -S(O) _₂C Substitution of _1-4 alkyl, -S(O)NH ₂ , -S(O)NHC _1-4 alkyl, -S(O)N(C _1-4 alkyl) ₂ , -S(O) _{2NH 2 ,} -S(O) _{2NHC 1-4} alkyl, -S(O) _2N (C _1-4 alkyl) ₂ groups;

And Cy is neither substituted nor unsubstituted.

or,

(4) A is * to indicate that it is connected to an alkyl carbon;

_R1 , _R2 , and _R3 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, _C3-6 cycloalkyl, and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O. The alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, and heterocycles are optionally substituted by 1-3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 , and -COOH, and _R1 , _R2 , and _R3 are not all H.

Optionally, _R1 and _R2 form a _C3-6 cycloalkyl group or a 4-7 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the cycloalkyl group or heterocycle is optionally substituted with 1-3 groups selected from =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 ynyl, and _C3-6 cycloalkyl.

As a second technical solution of the present invention, the compound of formula (I), its stereoisomer, deuterated product, eutectic, solvate, or pharmaceutically acceptable salt, has the structure of formula (II):

Each group is as described in the first technical solution.

As a third technical solution of the present invention, the compound of formula (II), its stereoisomer, deuterated product, eutectic, solvate, or pharmaceutically acceptable salt, wherein,

Ring G is a 5-9 member monocyclic carbon ring, a 5-7 member monocyclic heterocycle containing 1-3 heteroatoms selected from N, S, and O, or a fused ring of formula (I-1). L ₁ replaces any hydrogen atom on the ring G and is connected. The monocyclic carbon ring or monocyclic heterocycle is optionally replaced by 1-3 R _G.

Optionally, equation (I-1) has a structure

Optionally, equation (I-1) has a structure

Each _RG is independently selected from deuterium, _SF5 , =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, halo- _C1-4 alkyl, _C1-4 alkoxy, _C3-6 cycloalkyloxy, _C3-6 cycloalkyl, _C2-6 alkenyl, _C2-6 alkynyl, _-NHC1-4 alkyl, -N( _C1-4 alkyl) ₂ , _-COC1-4 alkyl, _-COOC1-4 alkyl, _-CONH2 , _-CONHC1-4 alkyl, -CONHC3-6 cycloalkyl, -CON( _C1-4 alkyl) _{2 , -NHCOC1-4} alkyl, _-NHCOC3-6 cycloalkyl, -P(O)( _C1-4 alkyl) ₂ , -S( _O ) _C1-4 alkyl, -S(O) _2C1-4 alkyl, -S(O) _2C The RG may be further substituted with 1-3 groups selected from N, S, and O heteroatoms, including _3-6 cycloalkyl, -S(O)NH ₂ , -S(O)NHC _1-4 alkyl, -S(O)N(C _1-4 alkyl) _2, -S(O) ₂ NH ₂ , and 4-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O heteroatoms. The _RG may also be further substituted with 1-3 groups selected from deuterium, C _1-4 alkyl, halo-C _1-4 alkyl, hydroxy-C _1-4 alkyl, -CONH ₂ , NH ₂ , C _1-6 alkoxy, hydroxyl, -COOH, halogen, and 5-7 membered heterocycles containing 1-3 heteroatoms selected from N, S, and O heteroatoms. Further, the 5-7 membered heterocycle may be substituted with 1-2 =O, halogen, cyano, C _1-4 alkyl, or halo-C _1-4 alkyl groups.

Cy is a heterocycle containing 1-3 heteroatoms selected from N, S, and O, consisting of 5-8 membered monocyclic heterocycles, 7-10 membered spirocyclic heterocycles, 6-9 membered bridged heterocycles, or 6-10 membered fused heterocycles. Cy is optionally surrounded by 1-3 heteroatoms selected from =O, halogen, cyano, hydroxyl, _NH₂ , -COOH, C₁ _-4 alkyl, halo-C₁ _-4 alkyl, hydroxy-C₁- ₄ alkyl, C₁ _-4 alkoxy, C₃ _-6 cycloalkyloxy, C₁ _-6 alkoxyalkyl, halo-C₁ _-4 alkoxy, -NHC₁- ₄ alkyl, -N(C₁ _-4 alkyl) _₂ , -COC₁ _-4 alkyl, -CONH₂, -CONHC₁ _{- 4} alkyl, -CON(C₁ _-4 alkyl) _₂ , -NHCOC₁ _-4 alkyl, -S(O)C₁ _-4 alkyl, -S(O) _₂C₂ Substitution of _1-4 alkyl, -S(O)NH ₂ , -S(O)NHC _1-4 alkyl, -S(O)N(C _1-4 alkyl) ₂ , -S(O) _{2NH 2 ,} -S(O) _{2NHC 1-4} alkyl, -S(O) _2N (C _1-4 alkyl) ₂ groups;

_R1 , _R2 and _R3 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl and _C2-6 alkynyl, wherein the alkyl, alkoxy, alkenyl and alkynyl groups are optionally substituted by 1 to 3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 and COOH;

Optionally, _R1 and _R2 form a _C3-6 cycloalkyl group or a 4-7 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the cycloalkyl group or heterocycle is optionally substituted by 1-3 groups selected from halogen, cyano, hydroxyl, _NH2 , COOH, and _C1-4 alkyl groups.

Each _R4 is independently selected from H, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, _C3-6 cycloalkyl, cyano, hydroxyl, _NH2 ; the alkyl, alkenyl, alkynyl, and cycloalkyl in _R4 are optionally substituted by 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, _NH2 , and COOH;

_R6 and _R7 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, _C2-6 alkynyl, and _C3-6 cycloalkyl; wherein the alkyl, alkoxy, alkenyl, alkynyl, and cycloalkyl groups are optionally substituted by 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, _NH2 , and COOH;

Alternatively, _R6 and _R7 form = 0;

Optionally, two _R5s on adjacent ring atoms, two _R6s on adjacent ring atoms, or _R5s and _R6s on adjacent ring atoms together with the atoms they are connected to form double bonds in _X1 , _X2 , _X3 and _X4 ;

Optionally, _R6 and _R7 on the same carbon atom form a _C3-6 cycloalkyl group or a 4-7 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the cycloalkyl group or heterocycle is optionally substituted by 1-3 groups selected from =O, halogen, cyano, hydroxyl, _NH2 , COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 ynyl and _C3-6 cycloalkyl groups;

The condition is that,

(1) A is a six-membered heteroaryl or a 5-7-membered non-aromatic monocyclic heterocycle, wherein the heteroaryl or non-aromatic monocyclic heterocycle contains 1-3 heteroatoms selected from N, S, and O, and A is optionally substituted by 1-3 R ₈ atoms;

* indicates the connection end with an alkyl carbon;

Ring B is _{a C4-6} carbon ring or a 5-6 membered heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the carbon ring or heterocycle is optionally substituted by 1-3 groups selected from =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C1-4 alkoxy, _-SC1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, and _C3-6 cycloalkyl.

Each R ₈ is independently selected from H, =O, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _-SC1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, cyano, hydroxyl, -COOH, NH2, and _C3-6 cycloalkyl; said alkyl, alkoxy, alkenyl, alkynyl, and cycloalkyl are optionally substituted by 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, _{NH2 ,} and -COOH;

And Cy is neither substituted nor unsubstituted.

or,

(2) A is a five-membered heteroaryl group containing 1-3 heteroatoms selected from N, S, and O, wherein A is optionally substituted by 1-3 R ₈ atoms;

Each R ₈ is independently selected from H, =O, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _-SC1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, cyano, hydroxyl, -COOH, NH2, and _C3-6 cycloalkyl; said alkyl, alkoxy, alkenyl, alkynyl, and cycloalkyl are optionally substituted by 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, _{NH2 ,} and -COOH;

And Cy is neither substituted nor unsubstituted.

or,

(3) A is * to indicate the connection end with the alkyl carbon;

_RA1 , _RA2 , _RA3 , and _RA4 are each independently _R8 , and _RA1 , _RA2 , _RA3 , and _RA4 are not simultaneously H;

Each R ₈ is independently selected from H, =O, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _-SC1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, cyano, hydroxyl, -COOH, NH2, and _C3-6 cycloalkyl; said alkyl, alkoxy, alkenyl, alkynyl, and cycloalkyl are optionally substituted by 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, _{NH2 ,} and -COOH;

Cy is a 5-8 membered monocyclic heterocycle containing 1-3 heteroatoms selected from N, S, and O, wherein the monocyclic heterocycle is optionally surrounded by 1-3 heteroatoms selected from =O, halogen, cyano, hydroxyl, _NH₂ , -COOH, _C1-4 alkyl, halo _{-C1-4 alkyl} , hydroxy- _C1-4 alkyl, _C1-4 alkoxy, C3-6 cycloalkyloxy, _C1-6 alkoxyalkyl, halo- _C1-4 alkoxy, _-NHC1-4 alkyl, -N( _C1-4 alkyl) _₂ , _-COC1-4 alkyl, -CONH₂, _-CONHC1-4 alkyl, -CON( _{C1-4 alkyl} ) _₂ , _-NHCOC1-4 alkyl, -S ₍ O) _C1-4 alkyl, -S(O) _₂C1-4 alkyl, -S(O) _NH₂ , -S(O)NHC Substitution of _1-4 alkyl, -S(O)N(C _1-4 alkyl) ₂ , -S(O) _{2NH 2 ,} -S(O) _{2NHC 1-4} alkyl, -S(O) _2N (C _1-4 alkyl) ₂ groups;

And Cy is neither substituted nor unsubstituted.

or,

(4) A is * to indicate that it is connected to an alkyl carbon;

_R1 , _R2 , and _R3 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, and _C2-6 alkynyl, wherein the alkyl, alkoxy, alkenyl, and alkynyl groups are optionally substituted by 1 to 3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 , and COOH, and _R1 , _R2 , and _R3 are not all H;

Optionally, _R1 and _R2 form a _C3-6 cycloalkyl group with the carbon atoms they are attached to, wherein the cycloalkyl group is optionally substituted with 1-3 groups selected from =O, halogen, cyano, hydroxyl, _NH2 , COOH, _C1-4 alkyl, _C2-6 alkenyl, and _C2-6 ynyl.

Undefined groups are as described in the second technical solution.

As a fourth technical solution of the present invention, the compound of formula (I) or formula (II), its stereoisomer, deuterated product, eutectic, solvate or pharmaceutically acceptable salt, wherein the compound further has the structure of formula (III):

Wherein, Rc is H, =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, halo- _C1-4 alkyl, hydroxy _-C1-4 alkyl, _C1-4 alkoxy, _C1-6 alkoxyalkyl, or halo- _C1-4 alkoxy.

_R1 , _R2 and _R3 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, wherein the alkyl or alkoxy group is optionally substituted by 1 to 3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 and COOH;

Optionally, _R1 and _R2 form a _C3-6 cycloalkyl group with the carbon atoms they are attached to, wherein the cycloalkyl group is optionally substituted by 1-3 groups selected from halogen, cyano, hydroxy, _NH2 , COOH, and _C1-4 alkyl groups;

L1 is a bond, _C1-3 alkylene, -NH-, -O-, -S-, _C2-6 alkenyl, _C2-6 alynyl, or -CO-, wherein the alkylene, alkenyl, or alynyl group is optionally substituted by 1-3 groups selected from halogen, cyano, hydroxyl, and _NH2 ;

Each R ₅ is independently selected from H, _C1-4 alkyl, and _C3-6 cycloalkyl; the alkyl or cycloalkyl group is optionally substituted with 1-3 groups selected from deuterium, halogen, cyano, hydroxyl, _NH2 , and COOH;

_R6 and _R7 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, and _C1-4 alkoxy; the alkyl and alkoxy groups are optionally substituted by 1 to 3 groups selected from deuterium, halogen, cyano, hydroxyl, _NH2 and COOH;

Alternatively, _R6 and _R7 form = 0;

Optionally, two _R5s on adjacent ring atoms, two _R6s on adjacent ring atoms, or _R5s and _R6s on adjacent ring atoms together with the atoms they are connected to form double bonds in _X1 , _X2 , _X3 and _X4 ;

The condition is that,

(1) A is a ring A that is optionally substituted by 1-3 R ₈ ; the substituents are located at any substituted position on the ring A, including on the carbon atom of the benzene ring and on the carbon atom or heteroatom of the ring fused with it;

Each _R8 is independently selected from halogen, =O, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl and _C3-6 cycloalkyl;

* indicates the connection end with an alkyl carbon;

r is an integer from 1 to 3;

E is selected from NH, S, and O;

or,

(2) A can be arbitrarily replaced by 1-3 R _8s ;

Each _R8 is independently selected from halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 alkynyl and _C3-6 cycloalkyl;

_Z1 and _Z2 are each independently CH or N;

_Z3 is S, O, or NH;

* indicates the connection end with an alkyl carbon;

or,

(3) A is * to indicate the connection end with the alkyl carbon;

_RA1 , _RA2 , _RA3 , and _RA4 are each independently _R8 , and _RA1 , _RA2 , _RA3 , and _RA4 are not simultaneously H;

Each R ₈ is independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, cyano, hydroxyl, _NH2 , COOH; the alkyl and alkoxy groups are optionally selected from 1 to 3 of deuterium, halogen, cyano, hydroxyl, _NH2 , and COOH;

or,

(4) A is * to indicate that it is connected to an alkyl carbon;

_R1 , _R2 , and _R3 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, and _C2-6 alkynyl, wherein the alkyl, alkoxy, alkenyl, and alkynyl groups are optionally substituted by 1 to 3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 , and COOH, and _R1 , _R2 , and _R3 are not all H;

Optionally, _R1 and _R2 form a _C3-6 cycloalkyl group with the carbon atoms they are attached to, wherein the cycloalkyl group is optionally substituted by 1-3 groups selected from halogen, cyano, hydroxyl, _NH2 , and COOH;

Undefined groups are as described in the third technical solution.

As the fifth technical solution of the present invention, the compound of formula (III), its stereoisomer, deuterated product, eutectic, solvate, or pharmaceutically acceptable salt, wherein,

Ring G can be cyclopentane, cyclohexane, cycloheptane, benzene ring, or ring G can be optionally substituted with 1-3 R _Gs .

Optionally, equation (I-2) has a structure

Optionally, equation (I-2) has a structure

_L1 replaces any hydrogen atom on the G ring and is used for connection.

Each _RG is independently selected from deuterium, _SF5 , =O, halogen, cyano, hydroxyl, _NH2 , _C1-4 alkyl, _C1-4 alkoxy, _C3-6 cycloalkyl, _C2-6 alkenyl, _C2-6 alkynyl, _-NHC1-4 alkyl, -N( _C1-4 alkyl) ₂ , _-COC1-4 alkyl, -CONH2, -CONHC1-4 alkyl, _{-CON(C1-4 alkyl)2, -NHCOC1-4 alkyl, -P(O)(C1-4 alkyl ) 2 , and 4-7} membered heterocycles containing 1-3 heteroatoms selected from N, S, and O, wherein the _RG is optionally further composed of 1-3 heteroatoms selected from deuterium, _C1-4 alkyl, halogenated _C1-4 alkyl, hydroxyl _C1-4 alkyl, _-CONH2 , _NH2 , C _{The 1-6} alkoxy, hydroxyl, halogen, and groups containing 1-3 5-7 membered heterocycles selected from N, S, and O heteroatoms are substituted; further, the 5-7 membered heterocycles are optionally substituted by 1-2 =O, halogen, cyano, _C1-4 alkyl, or halo _-C1-4 alkyl groups.

The condition is that,

(1) A is a ring A that is optionally substituted by 1-3 R ₈ ; the substituents are located at any substituted position on the ring A, including on the carbon atom of the benzene ring and on the carbon atom or heteroatom of the ring fused with it;

Each _R8 is independently selected from =O, halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 ynyl and _C3-6 cycloalkyl;

* indicates the connection end with an alkyl carbon; or

(2) A is optionally substituted with 1-3 groups selected from halogen, cyano, hydroxyl, _NH2 , -COOH, _C1-4 alkyl, _C2-6 alkenyl, _C2-6 ynyl and _C3-6 cycloalkyl;

* indicates the connection end with an alkyl carbon;

or,

(3) A is * to indicate the connection end with the alkyl carbon;

_RA1 , _RA2 , _RA3 , and _RA4 are each independently _R8 , and _RA1 , _RA2 , _RA3 , and _RA4 are not simultaneously H;

Each R ₈ is independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, cyano, hydroxyl, _NH2 , COOH; the alkyl and alkoxy groups are optionally selected from 1 to 3 of deuterium, halogen, cyano, hydroxyl, _NH2 , and COOH;

or,

(4) A is * to indicate that it is connected to an alkyl carbon;

_R1 , _R2 , and _R3 are each independently selected from H, deuterium, halogen, _C1-4 alkyl, _C1-4 alkoxy, _C2-6 alkenyl, and _C2-6 alkynyl, wherein the alkyl, alkoxy, alkenyl, and alkynyl groups are optionally substituted by 1 to 3 groups selected from halogen, _C1-4 alkyl, cyano, hydroxyl, _NH2 , and COOH, and _R1 , _R2 , and _R3 are not all H;

Optionally, _R1 and _R2 form a _C3-6 cycloalkyl group with the carbon atoms they are attached to, said cycloalkyl group being optionally substituted with 1-3 groups selected from halogen, cyano, hydroxyl, _NH2 , and COOH.

Undefined groups are as described in the fourth technical solution.

As the sixth technical solution of the present invention, the compound of formula (III), its stereoisomer, deuterated product, eutectic, solvate or pharmaceutically acceptable salt, wherein it is selected from one of the following structures:

Undefined groups are as described in the fifth technical solution.

As the seventh technical solution of the present invention, the compound of formula (I), its stereoisomer, deuterated derivative, eutectic, solvate, or pharmaceutically acceptable salt, wherein the compound has the structure of formula (IV):

Wherein, Rc is H, halogen, _C1-2 alkyl or _C1-2 alkoxy;

_R1 , _R2 and _R3 are each independently selected from H, deuterium, F, Cl, Br, methyl, ethyl, methoxy or ethoxy, wherein the methyl, ethyl, methoxy or ethoxy group is optionally substituted by 1 to 3 groups selected from F, Cl, Br, cyano, hydroxyl and _NH2 ;

Z is CH or N;

Ring G is a benzene ring optionally substituted with 1-2 _RGs or

Each R _G is independently selected from F, Cl, Br, I, methyl, ethyl, propyl, _SF5 and CN, wherein the methyl, ethyl or propyl group is optionally further substituted by 1 to 3 groups selected from deuterium, F, Cl, Br and I.

As the eighth technical solution of the present invention, the compound of formula (I), its stereoisomer, deuterated product, eutectic, solvate, or pharmaceutically acceptable salt, wherein the compound is selected from one of the following structures:

As the ninth technical solution of the present invention, the compound of formula (I) is a stereoisomer, deuterated product, eutectic, solvate, or pharmaceutically acceptable salt, wherein the compound is selected from one of the following structures:

As the tenth technical solution of the present invention, the present invention also provides a pharmaceutical composition comprising the compound described in any one of the first to ninth technical solutions, its stereoisomer, deuterated product, eutectic, solvate or pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier and/or excipient.

The present invention further provides the use of the compounds described in any one of the first to ninth technical solutions, their stereoisomers, deuterated derivatives, eutectics, solvates or pharmaceutically acceptable salts, or the compositions described in the tenth technical solution, in the preparation of medicaments for treating dipeptidyl peptidase 1-mediated diseases.

Furthermore, the diseases mediated by dipeptidyl peptidase 1 are selected from airway obstructive diseases, bronchiectasis, cystic fibrosis, asthma, emphysema, and chronic obstructive pulmonary disease, etc.

Synthetic route

Those skilled in the art can prepare the compounds of this invention using known organic synthesis techniques, with starting materials being commercially available chemicals and/or compounds described in chemical literature. "Commercially available chemicals" are obtained from legitimate commercial sources, and suppliers include: Titan Technology, Energie Chemicals, Shanghai Demo, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, Nanjing Yaoshi, WuXi AppTec, and Bailingwei Technology, among others.

References and monographs in this field provide detailed descriptions of the synthesis of reactants that can be used to prepare the compounds described herein, or provide articles describing such preparation methods for reference. These reference books and monographs include: "Synthetic Organic Chemistry", John Wiley & Sons, Inc., New York; S.R. Sandler et al., "Organic Functional Group Preparations," 2nd Ed., Academic Press, New York, 1983; H.O. House, "Modern Synthetic Reacti ons", 2nd Ed., W.A. Benjamin, Inc. Menlo Park, Calif. 1972; T.L. Gilchrist, "Heterocyclic Chemistry", 2nd Ed., John Wiley & Sons, New York, 1992; J. March, "Advanced Organic Chemistry: Reactions, Mechanisms and Structure", 4th Ed., Wiley-Interscience, New York, 1992; Fuhrhop, J. and Penzlin G. "Organic Synthesis: Concepts, Methods, Starting Materials", Second, Revised and Enlarged Edition (1994) John Wiley & Sons ISBN: 3-527-29074-5; Hof fman, R.V. "Organic Chemistry, An Intermediate Text" (1996) Oxford University Press, ISBN 0-19-509618-5; Larock, R.C. "Comprehensive Organic Transformations: A Guide to Functional Group Preparations" 2nd Edition ( 1999) Wiley-VCH, ISBN: 0-471-19031-4; March, J. "Advanced Organic Chemistry: Reactions, Mechanisms, and Stru cture" 4th Edition (1992) John Wiley & Sons, ISBN: 0-471-60180-2; Otera, J. (editor) "ModernCarbonyl Chemistry" ( 2000) Wiley-VCH, ISBN: 3-527-29871-1; Patai, S. "Patai’s 1992Guide to the Chemistry of Functional Groups" (1992) Interscience ISBN: 0-471-93022-9; Solomons, T.W.G. "Organic Chemistry" 7th Edition (2000) John Wiley& Sons, ISBN: 0-471-19095-0; Stowell, J.C., "Intermediate Organic Chemistry" 2nd Edition (1993) Wiley-Interscience, ISBN: 0-471-57456-2; "Industrial Organic Chemicals: Starting Materials and Intermediates: An Ullma nn’s Encyclopedia” (1999) John Wiley & Sons, ISBN: 3-527-29645-X, in 8 volumes; “Organic Reactions” (1942-2000 ) John Wiley & Sons, in over 55 volumes; and “Chemistry of Functional Groups” John Wiley & Sons, in 73 volumes.

Specific and similar reactants can be selectively identified using indexes of known chemical substances prepared by the American Chemical Society's Chemical Abstracts Service. These indexes are available in most public and university libraries, as well as online. Chemicals known but not commercially available in the catalogue can optionally be prepared by custom chemical synthesis plants, many of which offer custom synthesis services (e.g., those listed above). A reference for the preparation and selection of pharmaceutical salts of the compounds described herein is P.H. Stahl & C.G. Wermuth, “Handbook of Pharmaceutical Salts”, Verlag Helvetica Chimica Acta, Zurich, 2002.

the term

Unless otherwise specified in this invention, the terminology used in this invention has the following meanings:

The carbon, hydrogen, oxygen, sulfur, nitrogen, or halogen involved in the groups and compounds described in this invention include their isotopes, and the carbon, hydrogen, oxygen, sulfur, nitrogen, or halogen involved in the groups and compounds described in this invention may optionally be further replaced by one or more of their corresponding isotopes, wherein the isotopes of carbon include ^12C , ^13C , and ^14C , the isotopes of hydrogen include protium (H), deuterium (also known as heavy hydrogen), and tritium (also known as superheavy hydrogen), the isotopes of oxygen include ^16O , ^17O , and ^18O , the isotopes of sulfur include ^32S , ^33S , ^34S , and ^36S , the isotopes of nitrogen include ^14N and ^15N , the isotope of fluorine is ^19F , the isotopes of chlorine include ^35Cl and ^37Cl , and the isotopes of bromine include ^79Br and ^81Br .

In this article, "halogen" refers to F, Cl, Br, I, or their isotopes.

"Halogenation" or "halogen substitution" refers to substitution by one or more halogen substituents selected from F, Cl, Br, I, or their isotopes. The upper limit of the number of halogen substituents is equal to the sum of the number of hydrogen atoms that can be substituted in the substituted group. Unless otherwise specified, the number of halogen substituents can be any integer between 1 and this upper limit. When the number of halogen substituents is greater than 1, the substitution can be by the same or different halogens. Common cases include 1-5 halogen substitutions, 1-3 halogen substitutions, 1-2 halogen substitutions, and 1 halogen substitution.

"Deuterium" refers to the hydrogen (H) isotope deuterium.

"Deuteration" refers to the situation where hydrogen atoms on alkyl, cycloalkyl, alkylene, aryl, heteroaryl, alkenyl, alkynyl, and other groups are replaced by at least one isotope, deuterium. The upper limit of the number of deuterations is equal to the sum of the number of hydrogen atoms that can be replaced in the substituted group. Unless otherwise specified, the number of deuterations is any integer between 1 and the upper limit, preferably 1-20 deuterium atoms, more preferably 1-10 deuterium atoms, more preferably 1-6 deuterium atoms, and even more preferably 1-3 deuterium atoms.

"C _xy " groups refer to groups containing x to y carbon atoms, such as "C _1-6 alkyl" which refers to alkyl groups containing 1 to 6 carbon atoms.

"alkyl" refers to a monovalent straight-chain or branched saturated aliphatic hydrocarbon group, and unless otherwise specified, it is an alkyl group with 1 to 20 carbon atoms, preferably an alkyl group with 1 to 8 carbon atoms, more preferably an alkyl group with 1 to 6 carbon atoms, and even more preferably an alkyl group with 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, neobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, and their various branched isomers.

"Alkylene" refers to divalent straight-chain and branched saturated alkyl groups. Examples of alkylene groups include, but are not limited to, methylene, ethylene, etc.

"Haloalkyl" refers to an alkyl group in which one or more hydrogen atoms are replaced by one or more halogen atoms (such as fluorine, chlorine, bromine, iodine, or their isotopes). The upper limit of the number of halogen substituents is equal to the sum of the number of hydrogen atoms that can be substituted in the alkyl group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and this upper limit. Typically, alkyl groups are substituted by 1-5 halogens, or 1-3 halogens, or 1-2 halogens, or 1 halogen. When the number of halogen substituents is greater than 1, they can be the same or different halogens. Specific examples include, but are not limited to, _-CF3 , _-CH2Cl , _-CH2CF3 , _-CCl2 , _CF3 , _etc.

"Alkoxy" or "alkyloxy" refers to -O-alkyl. For example, -OC _1-8 alkyl, -OC _1-6 alkyl, -OC _1-4 alkyl, or -OC _1-2 alkyl. Specific non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexoxy, cyclopropoxy, and cyclobutoxy, etc.; the alkoxy group may optionally be substituted with a substituent.

"Haloalkoxy" refers to -O-haloalkyl. For example, -O- _haloC1-8 alkyl, -O- _haloC1-6 alkyl, -O- _haloC1-4 alkyl, or -O- _haloC1-2 alkyl; the upper limit of the number of halogen substituents is equal to the sum of the number of hydrogens that can be substituted in the substituted group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and this upper limit, preferably 1-5 halogen substituents, 1-3 halogen substituents, 1-2 halogen substituents, or 1 halogen substituent; when the number of halogen substituents is greater than 1, the same or different halogens can be used for substitution; non-limiting examples include monofluoromethoxy, difluoromethoxy, trifluoromethoxy, difluoroethyloxy, etc.

"alkylamino" or "alkylamino" refers to an amino group substituted with one or two alkyl groups, also written as -N-(alkyl) ₂ or -NH-alkyl, the latter also written as monoalkylamino. Non-limiting examples include dimethylamino, monomethylamino, diethylamino, monoethylamino, etc.

"Alkenyl" refers to a straight-chain or branched hydrocarbon group containing at least one carbon-carbon double bond (C=C), typically containing 2 to 18 carbon atoms, such as 2 to 8 carbon atoms, further such as 2 to 6 carbon atoms, and even further such as 2 to 4 carbon atoms. Examples include, but are not limited to, vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2-methyl-1-butenyl, 2... -Methyl-3-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 1-octenyl, 3-octenyl, 1-nonenyl, 3-nonenyl, 1-decenyl, 4-decenyl, 1,3-butadiene, 1,3-pentadiene, 1,4-pentadiene, and 1,4-hexadiene, etc.; the alkenyl group may optionally be further substituted with substituents.

"Alkenyl" refers to a straight-chain or branched divalent unsaturated hydrocarbon group containing at least one carbon-carbon double bond (C=C). Unless otherwise specified, alkenyl contains 2-6 carbon atoms, preferably 2-4 carbon atoms. Non-limiting examples include alkenylene. The alkenyl group may optionally be substituted with substituents.

"Alynyl" refers to a straight-chain or branched hydrocarbon group containing at least one carbon-carbon triple bond (C≡C), typically containing 2 to 18 carbon atoms, further containing 2 to 8 carbon atoms, further containing 2 to 6 carbon atoms, and further containing 2 to 4 carbon atoms. Examples include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, butynyl, 2-butynyl, 3-butynyl, 1-methyl-2-propynyl, 4-pentynyl, 3-pentynyl, 1-methyl-2-butynyl, 2-hexynyl, 3-hexynyl, 2-hepynyl, 3-hepynyl, 4-hepynyl, 3-octyynyl, 3-nonynyl, and 4-decynyl; the alkynyl group may optionally be substituted with substituents.

"Alynyl" refers to a straight-chain or branched divalent unsaturated hydrocarbon group containing a carbon-carbon triple bond (C≡C), typically containing 2-6 carbon atoms, and more commonly 2-4 carbon atoms. Non-limiting examples include ethynyl, propynyl, and butynyl, wherein the ethynyl group may optionally be substituted with substituents. "Cycloalkyl" refers to a saturated or partially unsaturated, non-aromatic carbocyclic hydrocarbon group that does not contain cyclic heteroatoms. Cycloalkyl groups can be monocyclic, bicyclic, or polycyclic. Bicyclic or polycyclic groups can be fused, spirocyclic, bridged, or combinations thereof. A bicyclic or polycyclic group may include one or more aromatic rings, but the ring system as a whole is not aromatic, and the linking site may be on an aromatic ring or a non-aromatic ring. Typically, cycloalkyl groups contain 3 to 20 carbon atoms, more commonly 3 to 8 carbon atoms, and even more commonly 3 to 6 carbon atoms; when monocyclic, they contain 3 to 15 carbon atoms, or 3 to 10 carbon atoms, or 3 to 8 carbon atoms, or 3 to 6 carbon atoms; when bicyclic or polycyclic, they contain 5 to 12 carbon atoms, or 5 to 11 carbon atoms, or 6 to 10 carbon atoms; non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, butenyl, cyclopentenyl, cyclohexenyl, etc., and the cycloalkyl group may optionally be substituted with substituents. "Cycloalkylene" refers to a divalent saturated, substituted or unsubstituted cycloalkyl group, and non-limiting examples include...

"Carbocyclic" or "carbocyclic group" refers to a substituted or unsubstituted, saturated or unsaturated, aromatic or non-aromatic carbocyclic group, including monocyclic carbocyclic rings, bicyclic bridged rings, bicyclic fused rings, bicyclic spirocyclic rings, and tricyclic or multi-ringed groups, etc., typically having 3 to 14 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 6 to 8 carbon atoms or 3 to 6 carbon atoms. In non-limiting embodiments, monocyclic carbocyclic rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or phenyl, etc., bicyclic bridged rings include, etc., bicyclic fused rings include, etc., bicyclic spirocyclic rings include, etc., and tricyclic rings include, etc.

"Heterocycle" or "heterocyclic group" refers to a substituted or unsubstituted, saturated or unsaturated, aromatic or non-aromatic ring. Unless otherwise specified, it contains 1 to 5 heteroatoms selected from N, O, or S, preferably 1 to 4 heteroatoms, more preferably 1 to 3 heteroatoms, including monocyclic heterocycles, bicyclic bridged heterocycles, bicyclic fused heterocycles, and bicyclic spirocyclic heterocycles, as well as heterocycles with three or more members. Preferably, it is a 3- to 15-membered heterocycle, more preferably a 4- to 14-membered heterocycle, even more preferably a 4- to 10-membered heterocycle, a 5- to 12-membered heterocycle, and even more preferably a 5- to 8-membered heterocycle or a 5- to 6-membered heterocycle. The heterocycle is preferably a saturated heterocycle, such as a 5- to 12-membered saturated heterocycle, and even more preferably a 5- to 8-membered saturated heterocycle, a 7-membered saturated heterocycle, or a 5- to 6-membered saturated heterocycle. The ring atoms N and S of the heterocyclic group can be oxidized to various oxidation states. Heterocyclic groups can be attached to heteroatoms or carbon atoms. Non-limiting examples include epoxyethyl, azirropropyl, oxacyclobutyl, azirrobutyl, 1,3-dioxopentyl, 1,4-dioxopentyl, 1,3-dioxhexane, piperazinyl, azirroheptyl, pyridinyl, furanyl, thiopheneyl, pyranyl, N-alkylpyrroleyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyridazinyl, imidazolyl, piperidinyl, piperinyl, morpholinyl, thiomorpholinyl, 1,3-dithial. Dihydrofuranyl, dihydropyranyl, dithiapentylyl, tetrahydrofuranyl, tetrahydropyrroleyl, tetrahydroimidazoyl, oxazolyl, dihydrooxazolyl, tetrahydrooxazolyl, tetrahydrothiazoyl, tetrahydropyranyl, benzimidazolyl, benzopyridyl, pyrrolopyridyl, benzodihydrofuranyl, azabicyclo[3.2.1]octyl, azabicyclo[5.2.0]nonyl, oxatricyclo[5.3.1.1]dodecyl, azaadamantyl and oxaspiro[3.3]heptyl, etc.

"Hypo-heterocyclic group" refers to a divalent heterocyclic group that is substituted or unsubstituted, saturated or unsaturated, aromatic or non-aromatic. Non-limiting examples include, etc.

"Aryl" refers to an aromatic group, including 5- and 6-membered monocyclic aromatic groups containing 0 to 4 N, S, and O heteroatoms, as well as polycyclic systems with at least one aromatic ring. The concept includes aromatic carbocyclic and heterocyclic rings, such as phenyl, pyrrole, furan, thiophene, thiazole, isothiazole, imidazole, triazole, tetraazole, pyrazole, oxazole, isoxazole, pyridine, pyrazine, pyridazine, and pyrimidine. Polycyclic aryl groups (tricyclic or bicyclic) include naphthalene, benzoxazole, benzodioxazole, benzothiazole, benzimidazole, benzothiophene, methylenedioxybenzene, quinoline, isoquinoline, naphthidine, indole, benzofuran, purine, benzofuran, denitropurine, or indene. Aryl groups with heteroatoms in their ring structure can also be called "aryl heterocycles," "heteroaryl groups," or "heterocyclic aromatic rings."

A "spirocyclic ring" is a polycyclic group that shares a single carbon atom (called a spiro atom) between rings. It can contain 0 or more double or triple bonds and 0 to 5 heteroatoms selected from N, O, S, P, Si, and their oxidation states. Spirocyclic rings are typically 6 to 14-membered, 6 to 12-membered, or 6 to 10-membered. Common spirocyclic rings include tri-spirotri- (representing a three-membered ring followed by a spirotri-membered ring), tri-spirotetra-, tri-spiropenta-, tri-spirohexa-, tetra-spirotetra-, tetra-spiropenta-, tetra-spirohexa-, tetra-spiropenta-, tetra-spirohexa-, penta-spiropenta-, or penta-spirohexa-. Non-limiting examples of spirocyclic rings include...

The spiroring can be optionally replaced by a substituent.

"Built rings" refer to polycyclic groups that share two adjacent ring atoms and a chemical bond. They can contain one or more double or triple bonds and can contain 0 to 5 heteroatoms selected from N, S, O, P, Si, and their oxidation states. Typically, fused rings are 5 to 20-membered, 5 to 14-membered, 5 to 12-membered, or 5 to 10-membered rings. Common fused ring types include trifused tetracyclic rings (representing fused rings formed by a three-membered and a four-membered ring; according to IUPC nomenclature, this could be a fused ring with either a three-membered or a four-membered ring as the base ring, and the same applies below), trifused pentacyclic rings, trifused hexacyclic rings, tetrafused tetracyclic rings, tetrafused pentacyclic rings, tetrafused hexacyclic rings, pentafused pentacyclic rings, pentafused hexacyclic rings, and hexafused hexacyclic rings. Non-limiting examples of fused rings include purines, quinolines, isoquinolines, benzopyrans, benzofurans, and benzothiophenes.

The cyclic ring can be optionally replaced by a substituent.

A "bridged ring" refers to two rings sharing two non-adjacent ring atoms and may contain one or more double or triple bonds. Bridged rings can contain 0 to 5 heteroatoms selected from N, S, O, P, Si, and their oxidation states. Typically, bridged rings have 5 to 20, 5 to 14, 5 to 12, or 5 to 10 ring atoms. Non-limiting examples of bridged rings include adamantane, etc.

Unless otherwise specified, "substitution" or "substituent" refers to any substitution that occurs at a position permitted by chemical theory, and the number of substituents conforms to the rules of chemical bonding. Exemplary substituents include, but are not limited to: _C1-6 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, _C3-8 heteroalkyl, _C5-12 aryl, 5-12 heteroaryl, hydroxyl, _C1-6 alkoxy, _C5-12 aryloxy, thiol, _C1-6 alkylthio, cyano, halogen, C1-6 alkylthiocarbonyl, _C1-6 alkylcarbamoyl, N-carbamoyl, nitro, silyl, sulfinyl, sulfonyl, sulfoxide, halo- _C1-6 alkyl, halo- _C1-6 alkoxy, amino, phosphonic acid, -CO₂ ( _C1-6 alkyl), -OC(＝ _O )( _C1-6 alkyl), _-OCO₂ ( _C1-6 alkyl), -C(＝O) _NH₂ , -C(＝O)N( _{C1-6 alkyl} ) _₂ , -OC(＝O)NH(C _1-6 alkyl), -NHC(＝O)(C _1-6 alkyl), -N(C _1-6 alkyl)C(＝O)(C _1-6 alkyl), -NHCO ₂ (C _1-6 alkyl), -NHC(＝O)N(C _1-6 alkyl) ₂ , -HC(＝O)NH(C _1-6 alkyl), -NHC(＝O)NH ₂ , -NHSO ₂ (C _1-6 alkyl), -SO ₂ N(C _1-6 alkyl) ₂ , -SO ₂ NH(C _1-6 alkyl), -SO ₂ NH ₂ , -SO ₂ C _1-6 alkyl, etc.

"Optional" or "optionally" means that the event or environment described below may but does not have to occur, and the description includes the possibility or possibility that the event or environment may or may not occur. For example, "optionally substituted F alkyl" means that the alkyl group may but does not have to be substituted with F, and the description includes the case where the alkyl group is substituted with F and the case where the alkyl group is not substituted with F.

When the substituents are optionally further substituted, they do not contain groups that cannot be formed according to common chemical knowledge.

When a group is selected from atoms or groups such as H or deuterium, if the group is optionally further substituted, it does not contain H or deuterium atoms that are further substituted.

"Pharmaceutically acceptable salt" means that the compound of the present invention retains the bioavailability and properties of a free acid or a free base, wherein the free acid is obtained by reacting with a non-toxic inorganic or organic base, and the free base is obtained by reacting with a non-toxic inorganic or organic acid.

"Pharmaceutical composition" means one or more of the compounds described herein or their stereoisomers, solvates, pharmaceutically acceptable salts or eutectics, mixed with other components, wherein the other components contain physiologically/pharmaceuticalally acceptable carriers and/or excipients.

"Carrier" refers to a system that does not cause significant stimulation to the organism and does not eliminate the biological activity and properties of the given compound, and can change the way the drug enters the human body and its distribution in the body, control the release rate of the drug, and deliver the drug to the target organ. Non-limiting examples include microcapsules and microspheres, nanoparticles, liposomes, etc.

"Excipient" refers to an agent that is not itself a therapeutic agent but is used as a diluent, excipient, binder, and/or medium to be added to a pharmaceutical composition to improve its disposal or storage properties or to allow or promote the formation of a unit dosage form of the compound or pharmaceutical composition for administration. As known to those skilled in the art, pharmaceutical excipients can provide a variety of functions and can be described as wetting agents, buffers, suspending agents, lubricants, emulsifiers, disintegrants, absorbents, preservatives, surfactants, colorants, flavoring agents, and sweeteners. Examples of pharmaceutical excipients include, but are not limited to: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, cellulose acetate, hydroxypropyl methyl cellulose, hydroxypropyl cellulose, microcrystalline cellulose and croscarmellose (e.g. croscarmellose sodium); (4) tragacanth gum powder; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn... Oils and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer’s solution; (19) ethanol; (20) pH buffer solution; (21) polyester, polycarbonate and/or polyanhydride; and (22) other non-toxic compatible substances used in pharmaceutical preparations.

"Stereoisomers" are isomers that are produced by different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, and conformational isomers.

"Solvate" refers to a substance formed by the combination of the compound of the present invention or its salt with a stoichiometric or non-stoichiometric solvent through intermolecular non-covalent forces. When the solvent is water, it is a hydrate.

"Co-crystal" refers to a crystal formed by the bonding of an active pharmaceutical ingredient (API) and a co-crystal form (CCF) through hydrogen bonds or other non-covalent bonds. Both API and CCF are solids at room temperature in their pure states, and a fixed stoichiometric ratio exists between the components. Co-crystal is a multi-component crystal, encompassing both binary co-crystals formed between two neutral solids and multi-component co-crystals formed between a neutral solid and a salt or solvate.

Detailed Implementation

The present invention will be described in detail below through embodiments. Unless otherwise specified, experimental methods under conventional conditions were used in the embodiments. The embodiments are provided to better illustrate the present invention, but should not be construed as limiting the invention to the examples given. Non-essential improvements and adjustments made to the implementation schemes by those skilled in the art based on the above description are still within the scope of protection of the present invention.

Detection methods

The structure of the compounds was determined by nuclear magnetic resonance (NMR) and/or mass spectrometry (MS). NMR shifts (δ) are given in units of 10⁻⁶ (ppm). NMR measurements were performed using Bruker Avance III 400 and Bruker Avance 300 NMR spectrometers in the following solvents: deuterated dimethyl sulfoxide (DMSO-d₆), deuterated chloroform (CDCl₃), and deuterated methanol (CD₃OD), with tetramethylsilane (TMS) as the internal standard.

MS determination was performed using (Agilent 6120B (ESI) and Agilent 6120B (APCI));

HPLC determinations were performed using an Agilent 1260DAD high-performance liquid chromatograph (Zorbax SB-C18 100×4.6mm, 3.5μM).

Thin-layer chromatography silica gel plates are Yantai Huanghai HSGF254 or Qingdao GF254. The silica gel plates used in thin-layer chromatography (TLC) are 0.15mm-0.20mm in diameter, and the silica gel plates used for thin-layer chromatography separation and purification are 0.4mm-0.5mm in diameter.

Column chromatography typically uses Yantai Huanghai silica gel with a mesh size of 200-300 as the carrier.

Abbreviation:

Burgess reagent: (methoxycarbonylaminosulfonyl)triethylammonium hydroxide, inner salt

Pd(dppf) _Cl₂ : 1,1'-bis(diphenylphosphino)ferrocene palladium(II) chloride

X-Phos: 2-Dicyclohexylphosphine-2,4,6-triisopropylbiphenyl

DMF: N,N-dimethylformamide

HATU: 2-(7-Azobenzotriazole)-N,N,N',N'-Tetramethylurea hexafluorophosphate

DIPEA: N,N-Diisopropylethylamine

LDA: Lithium diisopropylamino

PE: Petroleum ether

EA: Ethyl acetate

THF: Tetrahydrofuran

MeOH: Methanol

DCM: Dichloromethane

TMSOTf: Trimethylsilyl trifluoromethanesulfonate

intermediate

INT-1: (S)-(1-cyano-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)phenyl)ethyl)tert-butyl carbamate

tert-butyl(S)-(1-cyano-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)ethyl)carbamate

Compound 1a (4.50 g, 12.1 mmol, preparation method according to WO2013041497) was dissolved in 1,4-dioxane (45 ml), and pinacolborane (3.12 g, 24.2 mmol), triethylamine (3.67 g, 36.3 mmol), and Pd(dppf) _Cl₂ (877 mg, 1.2 mmol) were added. After addition, the mixture was reacted in a microwave oven at 100 °C for 1 hour. After the reaction was complete, the mixture was filtered, the filtrate was concentrated, and the residue was purified by silica gel column chromatography (eluting ratio PE:EA(v/v) = 1:0 to 10:1) to give INT₁, a white solid (2.1 g, yield 46.7%). LCMS m/z = 373.2 [M+1] ^⁺ .

¹ H NMR (400MHz, CDCl ₃ ) δ7.81(d,2H),7.29(d,2H),4.81(s,1H),4.72(s,1H),3.15-3.03(m,2H),1.44(s,9H),1.34(s,12H).

INT-2: (S)-(1-cyano-2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)phenyl)ethyl)tert-butyl carbamate

tert-butyl(S)-(1-cyano-2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)ethyl)carbamate

Compound 2a (2.0 g, 5.8 mmol, preparation method according to WO2016016242) was dissolved in ethylene glycol dimethyl ether (40 ml), and pinacol diboron ester (2.23 g, 8.7 mmol), potassium acetate (1.70 g, 17.4 mmol), and Pd(dppf) _Cl₂ (423.1 mg, 0.58 mmol) were added. After the addition was complete, the mixture was heated to 90 °C and reacted for 3 hours. After the reaction was completed, the mixture was filtered, the filtrate was concentrated, and the residue was purified by silica gel column chromatography (eluting ratio PE:EA(v/v) = 1:0 to 10:1) to give INT₂, a white solid (2.1 g, yield 92.1%). LCMS m/z = 335.2 [M⁺¹-56] ^⁺ .

¹ H NMR (400MHz, CDCl ₃ ) δ7.56(d,1H),7.50(d,1H),7.29(d,1H),4.99(d,1H),4.82(s,1H),3.18-3.16(m,2H),1.42(s,9H),1.34(s,12H).

Example 1: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 1)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 1)

Step 1: (S)-(1-cyano-2-(2-fluoro-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)tert-butyl carbamate (1B)

(S)-tert-butyl(1-cyano-2-(2-fluoro-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)carbamate(1B)

1A (0.29 g, 0.85 mmol, synthetic method referred to WO 2016016242A1) was dissolved in 1,4-dioxane (10 mL) and water (0.4 mL). Intermediate 2a (0.35 g, 1.27 mmol), potassium carbonate (0.24 g, 1.70 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloromethane dichloride complex (70 mg, 0.09 mmol) were added. The mixture was reacted at 90 °C for 3 h. After cooling to room temperature, saturated sodium chloride aqueous solution (20 mL) was added, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 4:1) to give title compound 1B (white solid, 0.34 g, 99.0%). LC-MS (ESI): m/z=412.1[M+H] ⁺ .

Step 2: (S)-2-amino-3-(2-fluoro-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)propionitrile (1C)

(S)-2-amino-3-(2-fluoro-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)propanenitrile(1C)

1B (0.34 g, 0.83 mmol) was dissolved in formic acid (5 mL), and the reaction was allowed to proceed overnight at room temperature. The solution was concentrated to dryness, and ethyl acetate (25 mL) was added. The pH was adjusted to approximately 8 by dropwise addition of saturated sodium bicarbonate solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product, title compound 1C (pale yellow solid, 0.21 g, 69.5%). LC-MS (ESI): m/z = 312.1 [M+H] ⁺ .

Step 3: (S)-2-(((S)-1-cyano-2-(2-fluoro-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (1D)

(S)-tert-butyl 2-(((S)-1-cyano-2-(2-fluoro-4-(3-methyl-2-oxo-2,3–dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(1D)

1C (0.21 g, 0.60 mmol) was dissolved in DMF (10 mL), and DIPEA (0.23 g, 1.80 mmol), HATU (0.34 g, 0.90 mmol), and INT-3 (0.22 g, 0.90 mmol, preparation method referred to WO2015110826) were added. After the addition was complete, the reaction was allowed to proceed overnight at room temperature. The reaction was quenched dropwise by adding saturated ammonium chloride aqueous solution, followed by adding (30 mL) of saturated sodium chloride aqueous solution. The mixture was extracted with ethyl acetate (25 mL), and the organic phase was washed with saturated sodium chloride aqueous solution (25 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the title compound 1D (pale yellow solid, 0.32 g, 99.0%), which was directly used in the next reaction. LC-MS (ESI): m/z = 483.1 [M-57+H] ⁺ .

Step 4: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 1)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 1)

1D (0.32 g, 0.59 mmol) was dissolved in formic acid (2.5 mL), and the mixture was reacted at 50 °C for 10 min after the addition was complete. The solution was concentrated to dryness, and ethyl acetate (20 mL) was added. The pH was adjusted to approximately 8 by dropwise addition of saturated sodium bicarbonate solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 20:1) to give title compound 1 (0.15 g, 58.0%). LC-MS (ESI): m/z = 439.1 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ )δ7.43–7.22(m,5H),7.12(d,1H),5.19(dd,1H),4.18–4.04(m,1H),4.05–3.95(m, 1H),3.78(m,1H),3.46(s,3H),3.41–3.17(m,3H),3.03–2.87(m,3H),1.88(m,2H).

Example 2: N-((S)-1-cyano-2-(2-methoxy-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 2)

N-((S)-1-cyano-2-(2-methoxy-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 2)

Step 1: 4-Bromo-1-(Bromomethyl)-2-methoxybenzene (2B)

4-bromo-1-(bromomethyl)-2-methoxybenzene(2B)

(2A) (4 g, 18.43 mmol) was dissolved in DCM (60 mL), CBr ₄ (9.09 g, 27.64 mmol) was added, and PPh ₃ (7.24 g, 27.64 mmol) was slowly added. The mixture was reacted at room temperature for 1 hour. PE/EA (v/v = 5:1, 12 mL) was added to the reaction mixture, stirred, filtered, and the filtrate was concentrated. The residue was purified by silica gel column chromatography (PE:EA (v/v) = 30:1 to 20:1) to give the title compound (2B), a white solid (3.9 g, 76% yield).

Step 2: (2S,5R)-2-(4-bromo-2-methoxybenzyl)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazine (2C)

(2S,5R)-2-(4-bromo-2-methoxybenzyl)-5-isopropyl-3,6-dimethoxy-2,5-dihydropyrazine(2C)

(R)-2-isopropyl-3,6-dimethoxy-2,5-dihydropyrazine (1 g, 5.43 mmol) was dissolved in THF (20 mL), purged with nitrogen, and cooled to -78 °C. Butyllithium (6.5 mmol, 2.5 M in toluene, 2.6 mL) was added, and the reaction was allowed to proceed for 1 h. Then, a solution of 2B (1.67 g, 5.97 mmol) in THF (8 mL) was added, and the reaction was continued at -78 °C for 2 h. The reaction was quenched with saturated _NH₄Cl solution. The system was extracted with EA (10 mL × 3), and the organic phases were combined. The organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was column-sected on silica gel (EA/PE (v/v) = 0%–6%) to give the title compound (2C), a colorless oil (1.82 g, 87% yield). LCMS m/z = 383.3 [M+1] ⁺

Step 3: Methyl (S)-2-amino-3-(4-bromo-2-methoxyphenyl)propionate (2D)

methyl(S)-2-amino-3-(4-bromo-2-methoxyphenyl)propanoate(2D)

2C (1.82 g, 4.75 mmol) was dissolved in acetonitrile (15 mL), and 1 M HCl (5 mL) was added. The mixture was reacted overnight at room temperature. The system was evaporated to dryness, and the system was neutralized to weakly alkaline with saturated sodium bicarbonate aqueous solution. EA (15 mL × 3) was added for extraction and separation. The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (DCM:MeOH (v/v) = 20:1–10:1) to give 2D (1.34 g, 97% yield). LCMS m/z = 288.1 [M+1] ⁺

Step 4: (S)-3-(4-bromo-2-methoxyphenyl)-2-((tert-butoxycarbonyl)amino)methyl propionate (2E)

methyl(S)-3-(4-bromo-2-methoxyphenyl)-2-((tert-butoxycarbonyl)amino)propanoate(2E)

2D (1.34 g, 4.69 mmol) was dissolved in DCM (20 mL), and triethylamine (1.3 mL) and di-tert-butyl dicarbonate (1.23 g, 5.63 mmol) were added. The mixture was reacted at room temperature for 3 h. Water was added, and the mixture was extracted and separated. The organic phases were combined and washed with saturated brine (10 mL). The solution was dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (DCM:MeOH (v/v) = 20:1–10:1) to give 2E, a colorless oil (0.7 g, yield 31%). LCMS m/z = 288.1 [M-boc+1] ⁺

Step 5: (S)-3-(4-bromo-2-methoxyphenyl)-2-((tert-butoxycarbonyl)amino)propionic acid (2F)

(S)-3-(4-bromo-2-methoxyphenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid(2F)

2E (0.7 g, 1.87 mmol) was dissolved in methanol (10 mL) and water (6 mL), and NaOH (1.50 g, 3.74 mmol) was added. The mixture was reacted at room temperature for 2 hours. The methanol was removed by vortexing, and the pH was adjusted to weakly acidic with dilute hydrochloric acid. EA (10 mL × 3) was added for extraction and separation. The organic phase was evaporated to dryness to give the crude product (2F) (0.65 g). LCMS m/z = 274.2 [M-boc+1] ⁺

Step 6: (S)-(1-amino-3-(4-bromo-2-methoxyphenyl)-1-oxopropane-2-yl)tert-butyl carbamate (2G)

tert-butyl(S)-(1-amino-3-(4-bromo-2-methoxyphenyl)-1-oxopropan-2-yl)carbamate(2G)

2F (0.65 g, 1.73 mmol), _NH₄Cl (0.74 g), and HATU (0.66 g, 1.73 mmol) were dissolved in DMF (15 mL), and DIPEA (1.15 mL) was added. The mixture was reacted overnight at room temperature. Water and EA were added, and the mixture was extracted separately. The organic phase was dried over anhydrous _{Na₂SO₄ and} concentrated to give crude product (2 g) (0.62 g). LCMS m/z = 273.1 [M-boc+1] ⁺

Step 7: (S)-(2-(4-bromo-2-methoxyphenyl)-1-cyanoethyl)tert-butyl carbamate (2H)

tert-butyl(S)-(2-(4-bromo-2-methoxyphenyl)-1-cyanoethyl)carbamate(2H)

2G (0.62 g, 1.66 mmol) was dissolved in DCM (10 mL), and Burgess reagent (0.79 g, 3.32 mmol) was added. The mixture was reacted overnight at room temperature. The system was concentrated and evaporated to dryness. The crude product was purified by silica gel column chromatography (EA/PE (v/v) = 0%-25%) to give 2H (0.5 g, 85% yield). LCMS m/z = 355.1 [M+1] ⁺

Step 8: (S)-(1-cyano-2-(2-methoxy-4-(3-methyl-2-oxy-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)tert-butyl carbamate (2I)

tert-butyl(S)-(1-cyano-2-(2-methoxy-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)carbamate(2I)

2H (0.5 g, 1.41 mmol), 1B (0.11 g, 1.41 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloromethane dichloride complex (115 mg, 0.14 mmol), and potassium carbonate (390 mg, 2.82 mmol) were dissolved in 1,4-dioxane (20 mL). The system was protected by nitrogen purging and reacted at 90 °C for 3 hours. The solution was concentrated and evaporated to dryness, dissolved in DCM, filtered through diatomaceous earth, and the filtrate was evaporated to dryness. The crude product was separated by silica gel column chromatography (EA/PE (v/v) = 0%-40%) to give 2I, a pale yellow solid (550 mg, yield 92%). LCMS m/z = 424.2 [M+1] ⁺

Step 9: (S)-2-amino-3-(2-methoxy-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)propionitrile (2J)

(S)-2-amino-3-(2-methoxy-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)propanenitrile(2J)

2I (135 mg, 0.32 mmol) was dissolved in formic acid (8 mL) and stirred overnight at room temperature. The system was diluted with DCM, and the pH was adjusted to weakly alkaline with saturated sodium bicarbonate solution. The DCM and water were dissolved in water, and the organic phase was dried and concentrated. The solution was then evaporated to dryness to obtain the crude product (2 J) (90 mg), which was used directly in the next reaction. LCMS m/z = 324.1 [M+1] ⁺

Step 10: (S)-2-(((S)-1-cyano-2-(2-methoxy-4-(3-methyl-2-oxy-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazacycloheptane-4-carboxylic acid tert-butyl ester (2K)

tert-butyl(S)-2-(((S)-1-cyano-2-(2-methoxy-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(2K)

2J (90 mg, 0.28 mmol) and INT-3 (82 mg, 0.34 mmol) were dissolved in DMF (5 mL), and HATU (0.12 g, 0.31 mmol) and DIPEA (0.2 mL) were added. The mixture was reacted overnight at room temperature. Water and EA were added to the reaction mixture, and the mixture was extracted separately. The organic phase was dried and concentrated to give a pale yellow oily crude product. 2K (90 mg) was obtained by MeOH/DCM (v/v) = 0-10%. LCMS m/z = 549.1 [M+1] ⁺

Step 11: (S)-N-((S)-1-cyano-2-(2-methoxy-4-(3-methyl-2-oxy-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 2)

(S)-N-((S)-1-cyano-2-(2-methoxy-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 2)

2K (90 mg, 0.16 mmol) was dissolved in formic acid (5 mL), and the reaction was carried out at room temperature for 4 hours. The system was diluted with DCM, and the pH was adjusted to weakly alkaline with saturated sodium bicarbonate solution. The DCM and water phases were dissolved in water, dried, concentrated, and evaporated to dryness to obtain the residue. The residue was purified by silica gel column chromatography (DCM:MeOH (v/v) = 50:1–5:1) to give title compound 2 (13 mg, yield 18%). LC-MS m/z = 451.2 [M+1] ⁺

¹ H NMR (400MHz, CDCl ₃ )δ7.49(d,1H),7.32-7.28(m,2H),7.14-7.12(m,2H),7.05(d,1H),5.13-5.06(m,1H),4.18-4.14(m,1H),4.07-4.01(m,1H),3.96(s, 3H),3.82–3.76(m,1H),3.46(s,3H),3.44-3.41(m,1H),3.27-3.16(m,3H),3.07-2.97(m,3H),2.93-2.87(q,1H),1.98-1.94(q,2H).

Example 3: N-((S)-1-cyano-2-(3-fluoro-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 3)

(S)-N-((S)-1-cyano-2-(3-fluoro-4-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 3)

Compound 3 was prepared from compound 3A using the same method as compound 2. LC-MS m/z = 439.2 [M+1] ⁺

¹ H NMR (400MHz, CDCl ₃ )δ7.43(t,1H),7.31-7.27(m,2H),7.21-7.17(m,2H),7.14(d,1H),5.21-5.15(m,1H),4.30-4.27(m,1H),4.08-4.02 (m,1H),3.84-3.77(m,1H),3.53-3.49(m,1H),3.44(s,3H),3.23-3.04(m,5H),2.04-2.00(m,3H),1.26-1.22(m,1H).

¹⁹ F NMR (376MHz, CDCl ₃ ) δ-117.13.

Example 4: (S)-N-((S)-1-cyano-2-(5-(1-methyl-2-oxoindoline-6-yl)thiophen-2-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 4)

(S)-N-((S)-1-cyano-2-(5-(1-methyl-2-oxoindolin-6-yl)thiophen-2-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 4)

Step 1: 6-Bromo-1-methylindoline-2-one (4B)

6-bromo-1-methylindolin-2-one(4B)

4A (5 g, 23.5 mmol) was dissolved in 200 mL of acetonitrile, and potassium carbonate (23 g, 94.32 mmol) and methyl iodide (2.96 mL, 47.16 mmol) were added. The mixture was heated to 70 °C and stirred overnight. The reaction solution was concentrated, extracted with DCM and water, and the organic phase was dried and concentrated. Separation was performed by column chromatography (PE:EA = 2:1 (v/v)) to give a brown solid 4B (1.8 g, 34%). LC-MS (ESI): m/z = 226.1 [M+H] ⁺ .

Step 2: 1-Methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)indoline-2-one (4C)

Methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-2-one(4C)

Compound 4B (1 g, 4.42 mmol), palladium acetate (200 mg, 0.88 mmol), potassium acetate (0.87 g, 8.84 mmol), 2-(dicyclohexylphosphino)-2,4,6-triisopropylbiphenyl (1.2 g, 2.65 mmol), and pinacol diboronate (1.35 g, 5.3 mmol) were mixed and dissolved in 50 mL of 1,4-dioxane. The mixture was heated to 95 °C under _N2 protection and reacted for 3 hours. The reaction was monitored by LC-MS until complete. The reaction solution was concentrated and separated by column chromatography (PE:EA (v/v) = 1:1) to give a pale yellow solid 4C (880 mg, 73%). LC-MS (ESI): m/z = 274.2 [M+H] ⁺ .

Step 3: (S)-(1-cyano-2-(5-(1-(1-methyl-2-oxoindoline-6-yl)thiophene-2-yl)ethyl)tert-butyl carbamate (4D)

tert-butyl-(S)-(1-cyano-2-(5-(1-methyl-2-oxoindolin-6-yl)thiophen-2-yl)ethyl)carbamate(4D)

N-[(1S)-2-(5-bromothiophene-2-yl)-1-cyanoethyl] tert-butyl carbamate (480 mg, 1.45 mmol), 4C (475 mg, 1.74 mmol), potassium carbonate (400 mg, 2.9 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (21 mg, 0.29 mmol) were mixed and dissolved in 5 mL of 1,4-dioxane and 0.1 mL of water. The mixture was microwaved to 120 °C and reacted for 1 hour. The reaction was monitored by LC-MS until complete. The reaction solution was concentrated and separated by column chromatography (PE:EA(v/v) = 1:1) to give a pale yellow solid 4D (175 mg, 33%). LC-MS (ESI): m/z = 398.2 [M+H] ⁺ .

Step 4: (S)-2-amino-3-(5-(1-methyl-2-oxoindoline-6-yl)thiophen-2-yl)propionitrile (4E)

(S)-2-amino-3-(5-(1-methyl-2-oxoindolin-6-yl)thiophen-2-yl)propanenitrile(4E)

3 mL of formic acid was added to compound 4D (175 mg, 0.44 mmol), and the mixture was stirred at room temperature for 3 hours. After the reaction was monitored by LC-MS until complete, the pH of the reaction solution was adjusted to 10 by adding saturated sodium carbonate solution, followed by DCM extraction. The organic phase was dried and concentrated to give product 4E (119 mg, 91%). LC-MS (ESI): m/z = 298.1 [M+H] ⁺ .

Step 5: (S)-2-(((S)-1-cyano-2-(5-(1-methyl-2-oxoindoline-6-yl)thiophene-2-yl)ethyl)carbamoyl)-1,4-oxazacycloheptane-4-carboxylic acid tert-butyl ester (4F)

tert-butyl-(S)-2-(((S)-1-cyano-2-(5-(1-methyl-2-oxoindolin-6-yl)thiophen-2-yl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(4F)

Compound 4E (119 mg, 0.4 mmol), INT-3 (98 mg, 0.4 mmol), HATU (180 mg, 0.48 mmol), and DIEA (0.13 mL, 0.8 mmol) were mixed and dissolved in DMF, and stirred overnight at room temperature. The reaction was monitored by LC-MS until complete. Water was added, followed by extraction with EA. The organic phase was dried and concentrated, and separated by column chromatography (PE:EA (v/v) = 1:1) to give a yellow solid 4F (165 mg, 78%). LC-MS (ESI): m/z = 525.2 [M+H] ⁺ .

Step 6: (S)-N-((S)-1-cyano-2-(5-(1-methyl-2-oxoindoline-6-yl)thiophen-2-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 4)

(S)-N-((S)-1-cyano-2-(5-(1-methyl-2-oxoindolin-6-yl)thiophen-2-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 4)

Compound 4F (145 mg, 0.28 mmol) was dissolved in 3 mL of formic acid solution and reacted at room temperature for 3 hours. The reaction was monitored by LC-MS until complete. The reaction solution was poured into a saturated sodium carbonate solution, the pH was adjusted to 10, and the mixture was extracted with EA. The organic phase was dried and concentrated, and purified by silica gel column chromatography (methanol:dichloromethane (v/v) = 1:100–1:10) to give compound 4 (45 mg, 38%). LC-MS (ESI): m/z = 425.2 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ): 8.67-8.69(m,1H),7.40-7.41(d,1H),7.18-7.28(m,3H),6.99-7.00(m,1H),4.94-5.03(m,1H),4.01-4.04(m,1H),3. 71-3.77(m,1H),3.55(s,2H),3.33-3.46(m,3H),3.16(s,3H),3.07-3.12(m,1H),2.61-2.81(m,4H),1.69-1.78(m,2H).

Example 5: ((S)-N-((S)-1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophene-2-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 5)

(S)-N-((S)-1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophen-2-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 5)

Step 1: (S)-3-(5-bromothiophene-2-yl)-2-((tert-butoxycarbonyl)amino)propionic acid (5B)

(S)-3-(5-bromothiophen-2-yl)-2-((tert-butoxycarbonyl)amino)propanoiccid(5B)

5A (10 g, 40.3 mmol) was dissolved in methanol (200 mL), triethylamine (12.2 g, 120.9 mmol) was added, followed by di-tert-butyl dicarbonate (10.5 g, 48.36 mmol). The mixture was reacted at room temperature for 2 hours. After concentration, the pH was adjusted to 6–7 with dilute hydrochloric acid (1 N). Extraction with dichloromethane was followed by concentration and purification by silica gel column chromatography (methanol:dichloromethane (v/v) = 1:100–1:10) to give the title compound (5B), a yellow solid (14 g, 99% yield). LCMS m/z = 350.23 [M+1] ⁺

Step 2: (S)-(1-amino-3-(5-bromothiophen-2-yl)-1-oxopropane-2-yl)tert-butyl carbamate (5C)

tert-butyl(S)-(1-amino-3-(5-bromothiophen-2-yl)-1-oxopropan-2-yl)carbamate(5C)

5B (4 g, 11.5 mmol) was dissolved in DMF (50 mL), and ammonium chloride (620 mg, 11.5 mmol), 2-(1H-benzotriazo-L-1-yl)-1,1,3,3-tetramethylurea tetrafluoroborate (4.4 mg, 13.8 mmol), and DIPEA (2.9 g, 23.0 mmol) were added. The mixture was reacted overnight at room temperature, and water (50 mL) was added. The mixture was extracted with ethyl acetate (50 mL × 3), washed three times with water (50 mL × 3), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (methanol:dichloromethane (v/v) = 1:100–1:10) to give the title compound (5C), a pale yellow solid (1.8 g, 45% yield). LC-MS m/z = 349.01 [M+1] ⁺

Step 3: (S)-(2-(5-bromothiophene-2-yl)-1-cyanoethyl)tert-butyl carbamate (5D)

tert-butyl(S)-(2-(5-bromothiophen-2-yl)-1-cyanoethyl)carbamate(5D)

5C (1.8 g, 5.2 mmol) was dissolved in dichloromethane (50 mL), and Burgess reagent (1.6 g, 6.2 mmol) was added under ice bath conditions. The reaction was carried out at room temperature for 2 hours, followed by the addition of water (50 mL). The mixture was extracted with ethyl acetate (50 mL × 3), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (PE:EA (v/v) = 1:10–1:5) to give the title compound (5D) as a pale yellow solid (1.4 g, 81% yield). LCMS m/z = 331.23 [M+1] ⁺

Step 4: (S)-(1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophene-2-yl)ethyl)tert-butyl carbamate (5E)

tert-butyl(S)-(1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophen-2-yl)ethyl)carbamate(5E)

5D (700 mg, 2.1 mmol) was dissolved in dioxane (10 mL), and 3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)benzo[d]oxazol-2(3H) (825 mg, 3.0 mmol), potassium carbonate (869 mg, 6.3 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (70 mg) were added. Water (2 mL) was then added, and the reaction was carried out under nitrogen protection at 100 °C for 12 hours. After concentration, the mixture was purified by silica gel column chromatography (PE:EA(v/v) = 1:5 to 1:1) to give the title compound (5E), a yellow solid (290 mg, yield 34%). LCMS m/z = 400.13 [M+1] ⁺

Step 5: ((S)-2-amino-3-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl]thiophen-2-yl)propionitrile (5F)

(S)-2-amino-3-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophen-2-yl)propanenitrile(5F)

5E (290 mg, 0.73 mmol) was dissolved in formic acid (5 mL) and reacted at room temperature for 3 hours. The pH was adjusted to 7–8 with saturated sodium carbonate aqueous solution, and the mixture was extracted with dichloromethane (50 mL × 3). The organic phases were combined, washed with saturated sodium chloride aqueous solution (50 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (methanol:dichloromethane (v/v) = 1:100–1:10) to give the title compound (5F) as a yellow solid (150 mg, 69% yield). LCMS m/z = 300.07 [M+1] ⁺

Step 6: (S)-2-(((S)-1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophene-2-yl)ethyl)carbamoyl)-1,4-oxazacycloheptane-4-carboxylic acid tert-butyl ester (5G)

tert-butyl(S)-2-(((S)-1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophen-2-yl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(5G)

5F (90 mg, 0.30 mmol) was dissolved in DMF (10 mL), and HATU (152 mg, 0.4 mmol), DIEA (116 mg, 0.9 mmol), and (S)-4-(tert-butoxycarbonyl)-1,4-oxaheptane-2-carboxylic acid (73 mg, 0.30 mmol) were added sequentially. The mixture was reacted at room temperature for 12 hours, and water (30 mL) was added. The mixture was extracted with ethyl acetate (30 mL × 3), and the organic phases were combined. The organic phases were washed with water (30 mL × 2), then with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (methanol:dichloromethane (v/v) = 1:100–1:10) to give the title compound (5G), a yellow solid (100 mg, yield 64%). LCMS m/z = 527.19 [M+1] ⁺

Step 7: (S)-N-((S)-1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophen-2-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 5)

(S)-N-((S)-1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophen-2-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 5)

5 g (100 mg, 0.19 mmol) was dissolved in formic acid (2 mL) and reacted at room temperature for 3 hours. The pH was adjusted to 7–8 with saturated sodium carbonate aqueous solution. The mixture was extracted with dichloromethane (30 mL × 3). The organic phases were combined, washed with saturated sodium chloride aqueous solution (50 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (methanol:dichloromethane (v/v) = 1:100–1:10) to give the title compound 5 (25 mg, yield 31%). LCMS M/Z (ESI): m/z = 427.14 [M+1] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ8.72(d,1H),7.52–7.44(m,1H),7.40–7.29(m,3H),7.01(d,1H),5.04–4.88(m,1H),4.11–3.98(m,1H),3.98–3.84(m,1H),3 .81–3.68(m,1H),3.47–3.38(m,2H),3.38(s,3H),3.36–3.32(m,1H),3.24–3.12(m,1H),2.94–2.62(m,3H),1.90–1.69(m,2H).

Example 6: (S)-N-((S)-2-(4-(7-acetamido-2,3-dihydro-1H-inden-4-yl)-2-fluorophenyl)-1-cyanoethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 6)

(S)-N-((S)-2-(4-(7-acetamido-2,3-dihydro-1H-inden-4-yl)-2-fluorophenyl)-1-cyanoethyl)-1,4-oxazepane-2-carboxamide(compound 6)

Step 1: (S)-(2-(4-(7-acetamido-2,3-dihydro-1H-inden-4-yl)-2-fluorophenyl)-1-cyanoethyl)tert-butyl carbamate (6B)

tert-butyl(S)-(2-(4-(7-acetamido-2,3-dihydro-1H-inden-4-yl)-2-fluorophenyl)-1-cyanoethyl)carbamate(6B)

Compound 6A (254.0 mg, 1.00 mmol, preparation method reference: Journal of Medicinal Chemistry, 2015, 58, 878–887), INT-2 (470.0 mg, 1.20 mmol), Pd(dppf) _Cl₂ (160.0 mg, 0.20 mmol), and potassium carbonate (280.0 mg, 2.00 mmol) were added to a single-necked flask, followed by 1,4-dioxane (10 mL) and water (0.4 mL). After purging with nitrogen three times, the reaction was carried out at 95 °C for 4 hours. After cooling to room temperature, the mixture was concentrated and purified by silica gel column chromatography (PE:EA(v/v) = 2:1) to give the title compound 6B as a white solid (360.0 mg, 82.3%). LC-MS (ESI): m/z = 381.1 [M-57+H] ^⁺ .

Step 2: (S)-N-(7-(4-(2-amino-2-cyanoethyl)-3-fluorophenyl)-2,3-dihydro-1H-inden-4-yl)acetamide (6C)

(S)-N-(7-(4-(2-amino-2-cyanoethyl)-3-fluorophenyl)-2,3-dihydro-1H-inden-4-yl)acetamide(6C)

6B (360.0 mg, 0.82 mmol) was dissolved in formic acid (5 mL) and reacted at 35 °C for 4 h. The reaction mixture was adjusted to alkaline with saturated potassium carbonate solution, extracted with ethyl acetate (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 6C, a pale yellow oil (270.0 mg, 97.6%), which was used directly in the next reaction. LC-MS (ESI): m/z = 338.2 [M+H] ⁺ .

Step 3: (S)-2-(((S)-2-(4-(7-acetamido-2,3-dihydro-1H-inden-4-yl)-2-fluorophenyl)-1-cyanoethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (6D)

tert-butyl(S)-2-(((S)-2-(4-(7-acetamido-2,3-dihydro-1H-inden-4-yl)-2-fluorophenyl)-1-cyanoethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(6D)

6C (270 mg, 0.80 mmol) was dissolved in DMF (5 mL), and INT-3 (235.2 mg, 0.96 mmol), HATU (364.8 mg, 0.96 mmol), and DIPEA (309.6 mg, 2.40 mmol) were added. The mixture was reacted overnight at room temperature. Water (20 mL) was added to the system, and the mixture was extracted with ethyl acetate (30 mL × 2). The extract was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (PE:EA (v/v) = 2:1) to give the title compound 6D as a white solid (370.0 mg, 82.0%). LC-MS (ESI): m/z = 563.3 [MH] ^-.

Step 4: (S)-N-((S)-2-(4-(7-acetamido-2,3-dihydro-1H-inden-4-yl)-2-fluorophenyl)-1-cyanoethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 6)

(S)-N-((S)-2-(4-(7-acetamido-2,3-dihydro-1H-inden-4-yl)-2-fluorophenyl)-1-cyanoethyl)-1,4-oxazepane-2-carboxamide(compound 6)

6D (370 mg, 0.66 mmol) was dissolved in formic acid (5 mL) and reacted at 35 °C for 4 h. The reaction mixture was adjusted to alkaline with saturated potassium carbonate solution, extracted with ethyl acetate (30 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 30:1) to give title compound 6 (90.0 mg, 30.0%).

¹ H NMR (400MHz, CDCl ₃ )δ7.85(d,1H),7.34(t,1H),7.19–7.13(m,3H),6.98(s,1H),5.21–5.15(m ,1H),4.09(q,1H),4.03–3.97(m,1H),3.79–3.73(m,1H),3.29(dd,1H),3. 24–3.17(m,2H),2.99(t,2H),2.96–2.85(m,5H),2.22(s,3H),2.15–2.07( m,2H),1.87–1.80(m,2H),1.61–1.50(m,2H).LC-MS(ESI):m/z＝465.2[M+H] ⁺ .

Example 7: (S)-N-((S)-1-cyano-2-(4-(1,1-dioxy-2,3-dihydrobenzo[b]thiophene-5-yl)-2-fluorophenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 7)

(S)-N-((S)-1-cyano-2-(4-(1,1-dioxido-2,3-dihydrobenzo[b]thiophen-5-yl)-2-fluorophenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 7)

Step 1: (S)-(1-cyano-2-(4-(1,1-dioxy-2,3-dihydrobenzo[b]thiophene-5-yl)-2-fluorophenyl)ethyl)tert-butyl carbamate (7B)

Tert-butyl(S)-(1-cyano-2-(4-(1,1-dioxido-2,3-dihydrobenzo[b]thiophen-5-yl)-2-fluorophenyl)ethyl)carbamate(7B)

Compound 7A (0.200 g, 0.81 mmol, preparation method according to EP3342765), INT-2 (0.316 g, 0.81 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (0.066 g, 0.081 mmol), and potassium carbonate (0.33 g, 2.43 mmol) were dissolved in 1,4-dioxane (10 mL), water (2 mL) was added, and the mixture was purged three times with nitrogen. The reaction was carried out at 90°C for 4 hours under a nitrogen atmosphere. The reaction solution was concentrated to dryness, dissolved in dichloromethane, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 5:1) to give title compound 7B, a white solid (0.25 g, 71.8%). LC-MS (ESI): m/z = 431.1 [M+H] ⁺ .

Step 2: (S)-2-amino-3-(4-(1,1-dioxy-2,3-dihydrobenzo[b]thiophene-5-yl)-2-fluorophenyl)propionitrile 4-methylbenzenesulfonate (7C)

(S)-2-amino-3-(4-(1,1-dioxido-2,3-dihydrobenzo[b]thiophen-5-yl)-2-fluorophenyl)propanenitrile 4-methylbenzenesulfonate(7C)

7B (0.25 g, 0.58 mmol) was dissolved in acetonitrile (5 mL), and p-toluenesulfonic acid (0.331 g, 1.74 mmol) was added. After the addition was complete, the mixture was reacted at room temperature for 16 h. The mixture was filtered, and the filter cake was rinsed once with acetonitrile (2 mL). The filter cake was then evaporated to dryness to give the title compound 7C, a white solid (0.240 g, 82.2%), which was used directly in the next step of the reaction.

Step 3: (S)-2-(((S)-1-cyano-2-(4-(1,1-dioxy-2,3-dihydrobenzo[b]thiophene-5-yl]-2-fluorophenyl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (7D)

tert-butyl(S)-2-(((S)-1-cyano-2-(4-(1,1-dioxido-2,3-dihydrobenzo[b]thiophen-5-yl)-2-fluorophenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(7D)

INT-3 (0.161 g, 0.66 mmol) was dissolved in dichloromethane (10 mL), followed by the addition of triethylamine (0.17 g, 1.32 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate (0.25 g, 0.66 mmol). After the addition was complete, the mixture was stirred at room temperature for 1 hour. Then, 7C (0.24 g, 0.477 mmol) was added, and the mixture was allowed to react overnight at room temperature. After the reaction was complete, the mixture was concentrated, and the crude 7D was used directly in the next reaction. LC-MS (ESI): m/z = 556.3 [MH] ^- .

Step 4: (S)-N-((S)-1-cyano-2-(4-(1,1-dioxy-2,3-dihydrobenzo[b]thiophene-5-yl)-2-fluorophenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 7)

(S)-N-((S)-1-cyano-2-(4-(1,1-dioxido-2,3-dihydrobenzo[b]thiophen-5-yl)-2-fluorophenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 7)

The crude compound 7D was dissolved in acetonitrile (10 mL), and p-toluenesulfonic acid (0.331 g, 1.74 mmol) was added. After the addition was complete, the mixture was reacted at room temperature for 16 h, concentrated to dryness, and ethyl acetate (25 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 20:1) to give the title compound 7 (110 mg, two-step yield 50.5%).

¹ H NMR (400MHz, CDCl ₃ )δ7.82(d,1H),7.65(d,1H),7.54(s,1H),7.42(d,1H),7.38(s,1H),7.32(d,1H),7.25 –7.19(m,1H),5.22–5.11(m,1H),4.14-4.10(m,1H),4.09–4.00(m,1H),3.82-3.76(m, ,1H),3.58–3.52(m,2H),3.50–3.42(m,2H),3.41–3.32(m,1H),3.24(t,2H),3.00–2.9 3(m,2H),1.90(d,2H),1.60–1.51(m,1H),1.39(dd,1H).LC-MS(ESI):m/z＝458.1[M+H] ⁺ .

Example 8: (S)-N-((S)-1-cyano-2-(3-fluoro-4'-(pentafluoro-16-thio)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 8)

(S)-N-((S)-1-cyano-2-(3-fluoro-4'-(pentafluoro-l6-sulfaneyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide

Step 1: (S)-(1-cyano-2-(3-fluoro-4'-(pentafluoro-16-thio)-[1,1'-biphenyl]-4-yl)ethyl)tert-butyl carbamate (8A)

tert-butyl(S)-(1-cyano-2-(3-fluoro-4'-(pentafluoro-l6-sulfaneyl)-[1,1'-biphenyl]-4-yl)ethyl)carbamate

INT-2 (0.69 g, 1.77 mmol) was dissolved in dioxane (30 mL), followed by the addition of (4-bromophenyl)sulfur pentafluoride (0.5 g, 1.77 mmol), potassium carbonate (0.24 g, 1.77 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (260 mg, 0.35 mmol), then water (6 mL). The reaction was carried out under nitrogen protection at 100 °C for 4 hours. After concentration, the mixture was purified by silica gel column chromatography (PE:EA (v/v) = 1:10 to 1:5) to give the title compound 8A (600 mg, 80% yield). LCMS m/z = 467.11 [M+1] ⁺

Step 2: (S)-2-amino-3-(3-fluoro-4'-(pentafluoro-16-thio)-[1,1'-biphenyl]-4-yl)propionitrile (8B)

(S)-2-amino-3-(3-fluoro-4'-(pentafluoro-l6-sulfaneyl)-[1,1'-biphenyl]-4-yl)propanenitrile

8A (0.6 g, 1.29 mmol) was dissolved in acetonitrile (20 mL), and p-toluenesulfonic acid (0.67 g, 3.87 mmol) was added. The mixture was reacted at 30 °C for 2 hours, followed by the addition of water (30 mL). The pH was adjusted to 7–8 with saturated sodium carbonate aqueous solution. The mixture was extracted with dichloromethane (50 mL × 3), washed with saturated brine (50 mL × 1), dried over anhydrous sodium sulfate, and concentrated to give the title compound 8B (0.4 g, 84% yield). LCMS m/z = 367.32 [M+1] ⁺

Step 3: (S)-2-(((S)-1-cyano-2-(3-fluoro-4'-(pentafluoro-16-thio)-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (8C)

tert-butyl(S)-2-(((S)-1-cyano-2-(3-fluoro-4'-(pentafluoro-l6-sulfaneyl)-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate

8B (200 mg, 0.55 mmol) was dissolved in DMF (10 mL), followed by the addition of HATU (250 mg, 0.66 mmol), DIPEA (260 mg, 2.02 mmol), and INT-3 (130 mg, 0.55 mmol). The reaction was carried out at room temperature for 12 hours. Water (30 mL) was added, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with water (30 mL × 2), and then washed with saturated brine (30 mL × 1). The mixture was dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (ethyl acetate: dichloromethane (v/v) = 1:10 to 1:5) to give the title compound 8C (250 mg, yield 76%). LCMS m/z = 594.18 [M+1] ⁺

Step 4: (S)-N-((S)-1-cyano-2-(3-fluoro-4'-(pentafluoro-16-thio)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 8)

(S)-N-((S)-1-cyano-2-(3-fluoro-4'-(pentafluoro-l6-sulfaneyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide

8C (250 mg, 0.42 mmol) was dissolved in acetonitrile (20 mL), p-toluenesulfonic acid (220 mg, 1.26 mmol) was added, and the mixture was reacted at 30 °C for 3 hours. The pH was adjusted to 7–8 with saturated sodium carbonate aqueous solution, and the mixture was extracted with dichloromethane (40 mL × 3). The organic phases were combined, washed with saturated sodium chloride aqueous solution (50 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 0.01:1–0.1:1) to give title compound 8 (80 mg, 38%).

¹ H NMR (400MHz, DMSO-d ₆ )δ8.70(d,1H),8.12–7.87(m,4H),7.67–7.57(m,2H),7.54–7.47(m,1H),5.15–4.85(m,1H),4.03–3.97(m,1H),3.90–3.80(m,1H),3.7 6–3.69(m,1H),3.35–3.28(m,2H),3.26–3.19(m,1H),3.12–3.01(m,1H),2.88–2.72(m,1H),2.67–2.52(m,2H),1.81–1.62(m,2H).LCMS m/z(ESI): m/z＝494.13[M+1] ⁺

Example 9: (S)-N-((S)-1-cyano-2-(3-fluoro-3'-(pentafluoro-16-thio)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 9)

(S)-N-((S)-1-cyano-2-(3-fluoro-3'-(pentafluoro-l6-sulfaneyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide

Step 1: (S)-(1-cyano-2-(3-fluoro-3'-(pentafluoro-16-thio)-[1,1'-biphenyl]-4-yl)ethyl)tert-butyl carbamate (9A)

tert-butyl(S)-(1-cyano-2-(3-fluoro-3'-(pentafluoro-l6-sulfaneyl)-[1,1'-biphenyl]-4-yl)ethyl)carbamate

INT-2 (0.69 g, 1.77 mmol) was dissolved in dioxane (30 mL), followed by the addition of (3-bromophenyl)sulfur pentafluoride (0.5 g, 1.77 mmol), potassium carbonate (0.24 g, 1.77 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (260 mg, 0.35 mmol), then water (6 mL). The reaction was carried out under nitrogen protection at 100 °C for 4 hours. After concentration, the mixture was purified by silica gel column chromatography (PE:EA (v/v) = 1:10 to 1:5) to give the title compound 9A (600 mg, 80% yield). LCMS m/z = 467.11 [M+1] ⁺

Step 2: (S)-2-amino-3-(3-fluoro-4'-(pentafluoro-16-thio)-[1,1'-biphenyl]-4-yl)propionitrile (9B)

(S)-2-amino-3-(3-fluoro-4'-(pentafluoro-l6-sulfaneyl)-[1,1'-biphenyl]-4-yl)propanenitrile

9A (0.6 g, 1.29 mmol) was dissolved in acetonitrile (20 mL), and p-toluenesulfonic acid (0.67 g, 3.87 mmol) was added. The mixture was reacted at 30 °C for 2 hours, followed by the addition of water (30 mL). The pH was adjusted to 7–8 with a saturated sodium carbonate aqueous solution. The mixture was extracted with dichloromethane (50 mL × 3), washed with saturated brine (50 mL × 1), dried over anhydrous sodium sulfate, and concentrated to give the title compound 9B (0.4 g, 84% yield). LCMS m/z = 367.32 [M+1] ⁺

Step 3: (S)-2-(((S)-1-cyano-2-(3-fluoro-3'-(pentafluoro-16-thio)-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (9C)

tert-butyl(S)-2-(((S)-1-cyano-2-(3-fluoro-3'-(pentafluoro-l6-sulfaneyl)-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate

9B (200 mg, 0.55 mmol) was dissolved in DMF (10 mL), followed by the addition of HATU (250 mg, 0.66 mmol), DIEA (260 mg, 2.02 mmol), and INT-3 (130 mg, 0.55 mmol). The reaction was carried out at room temperature for 12 hours. Water (30 mL) was added, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with water (30 mL × 2), and then washed with saturated brine (30 mL × 1). The mixture was dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (ethyl acetate: dichloromethane (v/v) = 1:10 to 1:5) to give the title compound 9C (250 mg, yield 76%). LCMS m/z = 594.18 [M+1] ⁺

Step 4: (S)-N-((S)-1-cyano-2-(3-fluoro-3'-(pentafluoro-16-thio)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 9)

(S)-N-((S)-1-cyano-2-(3-fluoro-3'-(pentafluoro-l6-sulfaneyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide

9C (250 mg, 0.42 mmol) was dissolved in acetonitrile (20 mL), p-toluenesulfonic acid (220 mg, 1.26 mmol) was added, and the mixture was reacted at 30 °C for 3 hours. The pH was adjusted to 7–8 with saturated sodium carbonate aqueous solution, and the mixture was extracted with dichloromethane (40 mL × 3). The organic phases were combined, washed with saturated sodium chloride aqueous solution (50 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 0.01:1–0.1:1) to give title compound 9 (78 mg, 37%).

¹ H NMR (400MHz, DMSO-d ₆ )δ8.69(d,1H),8.12(s,1H),8.01(d,1H),7.93(d,1H),7.76–7.44(m,4H),5.21–4.95(m,1H),4.05–3.97(m,1H),3.91–3.80(m,1H), 3.77–3.63(m,1H),3.32(d,2H),3.26–3.18(m,1H),3.10–3.00(m,1H),2.83–2.73(m,1H),2.66–2.54(m,2H),1.81–1.61(m,2H).LCMS m/z(ESI): m/z＝494.13[M+1] ⁺

Example 10: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-oxo-1,2,3,4-tetrahydroisoquinoline-6-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 10)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

Step 1: (S)-(1-cyano-2-(2-fluoro-4-(1-oxo-1,2,3,4-tetrahydroisoquinoline-6-yl)phenyl)ethyl)tert-butyl carbamate (10A)

tert-butyl(S)-(1-cyano-2-(2-fluoro-4-(1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)phenyl)ethyl)carbamate

4A (0.27 g, 1.2 mmol), INT-2 (0.36 g, 0.92 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (71 mg, 0.1 mmol), and potassium carbonate (0.28 g, 2.0 mmol) were added sequentially to 1,4-dioxane (15 mL) and water (3 mL). The system was purged three times with nitrogen and reacted at 100 °C for 2 hours. After the reaction was complete, the mixture was cooled to room temperature, and water (50 mL) was added. The aqueous phase was extracted with ethyl acetate (50 mL × 2). The combined organic phases were washed with saturated sodium chloride aqueous solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by silica gel column chromatography (PE:EA = 3:1 to 1:1) to give 10A, a brown solid (0.32 g, yield 84%). LCMS m/z = 410.2 [M+1] ⁺ .

Step 2: (S)-2-amino-3-(2-fluoro-4-(1-oxo-1,2,3,4-tetrahydroisoquinoline-6-yl)phenyl)propionitrile (10B)

(S)-2-amino-3-(2-fluoro-4-(1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)phenyl)propanenitrile

10A (0.32 g, 0.78 mmol) was dissolved in formic acid (5.0 mL), and the mixture was reacted at 50 °C for 10 minutes after the addition was complete. The solution was concentrated to dryness, and ethyl acetate (60 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated, and the aqueous phase was extracted with ethyl acetate (60 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 10B (0.24 g, 100% yield) LCMS m/z = 310.2 [M+1] ⁺ .

Step 3: (S)-2-(((S)-1-cyano-2-(2-fluoro-4-(1-oxo-1,2,3,4-tetrahydroisoquinoline-6-yl)phenyl)ethyl)carbamoyl)-1,4-oxazacycloheptane-4-carboxylic acid tert-butyl ester (10C)

tert-butyl(S)-2-(((S)-1-cyano-2-(2-fluoro-4-(1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)phenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate

10B (0.24 g, 0.78 mmol) was dissolved in DMF (10 mL), and INT-3 (0.25 g, 1.0 mmol), diisopropylethylamine (0.19 g, 1.5 mmol), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (0.38 g, 1 mmol) were added. The mixture was reacted at room temperature for 1 hour after the addition was complete. Saturated sodium chloride aqueous solution (30 mL) was added, and the mixture was extracted with ethyl acetate (60 mL × 2). The organic phase was washed with saturated sodium chloride aqueous solution (60 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated by silica gel column chromatography (PE:EA = 2:1 to 1:2) to give the title compound 10C, a pale yellow solid (0.25 g, yield 60%).

Step 4: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-oxo-1,2,3,4-tetrahydroisoquinoline-6-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 10)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

10C (0.25 g, 0.47 mmol) was dissolved in formic acid (5.0 mL) and reacted at 50 °C for 10 min. The mixture was concentrated under reduced pressure, and ethyl acetate (60 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with ethyl acetate (60 mL × 5). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 4:1) to give compound 10 (0.12 g, yield 59%).

^1H NMR (400MHz, DMSO-d6 ₎ )δ8.78(d,1H),7.96–7.87(m,2H),7.70–7.64(m,2H),7.63–7.53(m,2H),7 .47(t,1H),5.11–5.01(m,1H),4.13-4.06(m,1H),3.92-3.82(m,1H),3.78 -3.70(m,1H),3.43–3.31(m,2H),3.26–3.10(m,2H),2.97(t,2H),2.94–2. 84(m,1H),2.79–2.61(m,2H),2.04-1.92(m,1H),1.82–1.73(m,2H).LC-MS m/z＝437.2[M+1] ⁺

Example 11: (S)-N-((S)-1-cyano-2-(4-(cyclopentylethynyl)-2-fluorophenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 11)

(S)-N-((S)-1-cyano-2-(4-(cyclopentylethynyl)-2-fluorophenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 11)

Step 1: (S)-(1-cyano-2-(4-(cyclopentylethynyl)-2-fluorophenyl)ethyl)tert-butyl carbamate (11B)

tert-butyl(S)-(1-cyano-2-(4-(cyclopentylethynyl)-2-fluorophenyl)ethyl)carbamate(11B)

Compound 11A (66 mg, 0.70 mmol), INT-2 (200 mg, 0.58 mmol), Pd(dppf) _Cl₂ (115.0 mg, 0.14 mmol), and potassium carbonate (193.2 mg, 1.4 mmol) were added to a single-necked flask, followed by 1,4-dioxane (10 mL) and water (0.4 mL). After purging with nitrogen three times, the reaction was carried out at 95 °C for 4 hours. After cooling to room temperature, the mixture was concentrated and purified by silica gel column chromatography (PE:EA(v/v) = 3:1) to give the title compound 11B as a white solid (172.0 mg, 83.5%). LC-MS (ESI): m/z = 300.1 [M-57+H] ^⁺ .

Step 2: (S)-2-amino-3-(4-(cyclopentylethynyl)-2-fluorophenyl)propionitrile (11C)

(S)-2-amino-3-(4-(cyclopentylethynyl)-2-fluorophenyl)propanenitrile(11C)

11B (172 mg, 0.48 mmol) was dissolved in formic acid (5 mL) and reacted at 35 °C for 4 h. The reaction mixture was adjusted to alkaline with saturated potassium carbonate solution, extracted with ethyl acetate (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 11C, a pale yellow oil (130 mg, 100.0%), which was used directly in the next reaction. LC-MS (ESI): m/z = 257.1 [M + H] ⁺ .

Step 3: (S)-2-(((S)-1-cyano-2-(4-(cyclopentylethynyl)-2-fluorophenyl)ethyl)carbamoyl)-1,4-oxazacycloheptane-4-carboxylic acid tert-butyl ester (11D)

tert-butyl(S)-2-(((S)-1-cyano-2-(4-(cyclopentylethynyl)-2-fluorophenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(11D)

11C (130 mg, 0.51 mmol) was dissolved in DMF (5 mL), and INT-3 (150 mg, 0.61 mmol), HATU (230 mg, 0.61 mmol), and DIPEA (200 mg, 1.53 mmol) were added. The mixture was reacted overnight at room temperature. Water (20 mL) was added to the system, and the mixture was extracted with ethyl acetate (30 mL × 2). The extract was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (PE:EA (v/v) = 2:1) to give the title compound 11D as a white solid (190.0 mg, 77.0%). LC-MS (ESI): m/z = 428.3 [M-57+H] ⁺ .

Step 4: (S)-N-((S)-1-cyano-2-(4-(cyclopentylethynyl)-2-fluorophenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 11)

(S)-N-((S)-1-cyano-2-(4-(cyclopentylethynyl)-2-fluorophenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 11)

11D (190 mg, 0.39 mmol) was dissolved in formic acid (5 mL) and reacted at 35 °C for 4 h. The reaction system was adjusted to alkaline with saturated potassium carbonate solution, extracted with ethyl acetate (30 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 30:1) to give compound 11 (50.0 mg, 33.0%).

¹ H NMR (400MHz, CDCl ₃ )δ7.20–7.17(m,2H),7.07(dd,1H),7.02(dd,1H),5.08–5.02(m,1H),4.09(q,1H),3.94–3.89(m,1H),3.71–3.64(m,1H),3.22(dd,1H),3.13 –3.03(m,2H),2.87–2.80(m,3H),2.78–2.70(m,1H),2.02–1.88(m,3H),1.82–1.67(m,4H),1.66–1.50(m,4H).LC-MS(ESI):m/z＝384.2[M+H] ⁺ .

Example 12: (S)-N-((S)-1-cyano-2-(4-(5-cyano-4-methylthiazo-2-yl)-2-fluorophenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 12)

(S)-N-((S)-1-cyano-2-(4-(5-cyano-4-methylthiazol-2-yl)-2-fluorophenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 12)

Step 1: 2-Bromo-4-methylthiazol-5-carboxamide (12B)

2-bromo-4-methylthiazole-5-carboxamide(12B)

12A (1.20 g, 4.8 mmol) was dissolved in 30% ammonia solution (30 mL) and reacted at 35 °C for 24 h. After cooling to room temperature, the mixture was extracted with ethyl acetate (30 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 12B as a yellow solid (0.80 g, 75.4%). LC-MS (ESI): m/z = 221.0 [M + H] ^⁺ .

Step 2: 2-Bromo-4-methylthiazolyl-5-onitrile (12C)

2-bromo-4-methylthiazole-5-carbonitrile(12C)

12B (0.55 g, 2.49 mmol) was dissolved in dichloromethane (25 mL), and Burgess reagent (1.19 g, 4.98 mmol) was added. The mixture was allowed to react overnight at room temperature. After concentration, the solution was purified by silica gel column chromatography (PE:EA (v/v) = 5:1) to give the title compound 12C as a white solid (0.40 g, 80.1%). LC-MS (ESI): m/z = 203.0 [M+H] ⁺ .

Step 3: (S)-(1-cyano-2-(4-(5-cyano-4-methylthiazolyl)-2-fluorophenyl)ethyl)tert-butyl carbamate (12D)

(S)-(1-cyano-2-(4-(5-cyano-4-methylthiazol-2-yl)-2-fluorophenyl)ethyl)tert-butyl carbamate(12D)

12C (199 mg, 0.98 mmol), INT-2 (458.9 mg, 1.18 mmol), Pd(dppf) _Cl₂ (160.5 mg, 0.20 mmol), and potassium carbonate (270.9 mg, 1.96 mmol) were added to a single-necked flask, followed by 1,4-dioxane (10 mL) and water (0.4 mL). After purging with nitrogen three times, the reaction was carried out at 95 °C for 4 hours. After cooling to room temperature, the mixture was concentrated and purified by silica gel column chromatography (PE:EA(v/v) = 3:1) to give the title compound 12D as a yellow solid (172.0 mg, 45.5%). LC-MS (ESI): m/z = 331.0 [M-57+H] ^⁺ .

Step 4: (S)-2-(4-(2-amino-2-cyanoethyl)-3-fluorophenyl)-4-methylthiazolyl-5-onitrile (12E)

(S)-2-(4-(2-amino-2-cyanoethyl)-3-fluorophenyl)-4-methylthiazole-5-carbonitrile(12E)

12D (172 mg, 0.45 mmol) was dissolved in formic acid (5 mL) and reacted at 35 °C for 4 h. The reaction mixture was adjusted to alkaline with saturated potassium carbonate solution, extracted with ethyl acetate (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 12E, a pale yellow oil (120 mg, 96.0%), which was used directly in the next reaction. LC-MS (ESI): m/z = 287.1 [M + H] ⁺ .

Step 5: (S)-2-(((S)-1-cyano-2-(4-(5-cyano-4-methylthiazolyl-2-yl)-2-fluorophenyl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (12F)

tert-butyl(S)-2-(((S)-1-cyano-2-(4-(5-cyano-4-methylthiazol-2-yl)-2-fluorophenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(12F)

12E (120 mg, 0.42 mmol) was dissolved in DMF (5 mL), and INT-3 (120 mg, 0.50 mmol), HATU (190 mg, 0.50 mmol), and DIPEA (160 mg, 1.26 mmol) were added. The mixture was reacted overnight at room temperature. Water (20 mL) was added to the system, and the mixture was extracted with ethyl acetate (30 mL × 2). The extract was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (PE:EA (v/v) = 2:1) to give the title compound 12F as a yellow solid (160.0 mg, 74.2%). LC-MS (ESI): m/z = 458.1 [M-57+H] ⁺ .

Step 6: (S)-N-((S)-1-cyano-2-(4-(5-cyano-4-methylthiazo-2-yl)-2-fluorophenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 12)

(S)-N-((S)-1-cyano-2-(4-(5-cyano-4-methylthiazol-2-yl)-2-fluorophenyl)ethyl)-1,4-oxazepane-2-carboxamide (Compound 12)

12F (160 mg, 0.31 mmol) was dissolved in formic acid (5 mL) and reacted at 35 °C for 4 h. The reaction mixture was adjusted to alkaline with saturated potassium carbonate solution, extracted with ethyl acetate (30 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 30:1) to give title compound 12 (50.0 mg, 39.0%).

¹ H NMR (400MHz, CDCl ₃ )δ7.72(dd,1H),7.69(dd,1H),7.44(t,1H),5.22–5.16(m,1H),4.09(q,1H),4.05–3.99(m,1H),3.80–3.73(m,1H), 3.30(dd,1H),3.26–3.23(m,2H),2.97–2.90(m,3H),2.67(s,3H),1.19–1.82(m,4H).LC-MS(ESI):m/z＝414.2[M+H] ⁺ .

Example 13: (S)-N-((S)-2-(4-(benzo[d]thiazo-2-yl)-2-fluorophenyl)-1-cyanoethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 13)

(S)-N-((S)-2-(4-(benzo[d]thiazol-2-yl)-2-fluorophenyl)-1-cyanoethyl)-1,4-oxazepane-2-carboxamide

Step 1: (S)-(1-cyano-2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-oxacyclopentaborane-2-yl)phenyl)ethyl)tert-butyl carbamate (13B)

tert-butyl(S)-(1-cyano-2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)ethyl)carbamate

INT-2 (5.00 g, 14.57 mmol) was dissolved in 1,4-dioxane (100 mL), and pinacol diboronate (4.81 g, 18.94 mmol), potassium acetate (4.29 g, 43.71 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (2.39 g, 2.91 mmol) were added. Under nitrogen protection, the mixture was heated to 100 °C for 2 h. After cooling to room temperature, the mixture was filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA(v/v) = 6:1) to give the title compound 13B, a colorless liquid (5.00 g, 87.93%). LC-MS (ESI): m/z = 391.2 [M+H] ⁺ .

Step 2: (S)-(2-(4-(benzo[d]thiazolyl-2-yl)-2-fluorophenyl)-1-cyanoethyl)tert-butyl carbamate (13C)

tert-butyl(S)-(2-(4-(benzo[d]thiazol-2-yl)-2-fluorophenyl)-1-cyanoethyl)carbamate

13B (0.40 g, 1.02 mmol) was dissolved in 1,4-dioxane (10 mL) and water (0.4 mL). 2-bromobenzothiazole (0.26 g, 1.22 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloromethane complex (0.17 g, 0.20 mmol), and potassium carbonate (0.28 g, 2.04 mmol) were added. Under nitrogen protection, the mixture was heated to 90 °C for 2 h. The reaction was allowed to proceed overnight at room temperature. The mixture was concentrated to dryness, and saturated ammonium chloride aqueous solution (50 mL) was added. Extraction was performed with ethyl acetate (20 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 4:1) to give the title compound 13C, a pale yellow liquid (0.24 g, 59.20%). LC-MS (ESI): m/z=398.1[M+H] ⁺ .

Step 3: (S)-2-amino-3-(4-(benzo[d]thiazolyl)-2-fluorophenyl)propionitrile (13D)

(S)-2-amino-3-(4-(benzo[d]thiazol-2-yl)-2-fluorophenyl)propanenitrile

13C (0.32 g, 0.81 mmol) was dissolved in formic acid (5 mL), and the reaction was carried out at 30 °C for 3 h after the addition was complete. The pH was adjusted to approximately 8 by dropwise addition of saturated sodium bicarbonate aqueous solution, and the mixture was extracted with ethyl acetate (25 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 13D as a colorless liquid (0.24 g, 99.65%), which was used directly in the next reaction. LC-MS (ESI): m/z = 298.1 [M+H] ⁺ .

Step 4: (S)-2-(((S)-2-(4-(benzo[d]thiazo-2-yl)-2-fluorophenyl)-1-cyanoethyl)carbamoyl)-1,4-oxazacycloheptane-4-carboxylic acid tert-butyl ester (13E)

tert-butyl(S)-2-(((S)-2-(4-(benzo[d]thiazol-2-yl)-2-fluorophenyl)-1-cyanoethyl)carbamoyl)-1,4-oxazepane-4-carboxylate

13D (0.24 g, 0.81 mmol) was dissolved in dichloromethane (10 mL), and INT-1 (0.26 g, 1.05 mmol), diisopropylethylamine (0.31 g, 2.43 mmol), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (0.46 g, 1.22 mmol) were added. The mixture was reacted overnight at room temperature. A saturated sodium chloride aqueous solution (30 mL) was added, and the mixture was extracted with ethyl acetate (25 mL). The organic phase was washed with saturated sodium chloride aqueous solution (25 mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 4:1) to give the title compound 13E, a pale yellow solid (0.25 g, 58.83%). LC-MS (ESI): m/z = 469.2 [M-57+H] ⁺ .

Step 5: (S)-N-((S)-2-(4-(benzo[d]thiazo-2-yl)-2-fluorophenyl)-1-cyanoethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 13)

(S)-N-((S)-2-(4-(benzo[d]thiazol-2-yl)-2-fluorophenyl)-1-cyanoethyl)-1,4-oxazepane-2-carboxamide

13E (0.32 g, 0.59 mmol) was dissolved in formic acid (2.0 mL), and the mixture was reacted at 35 °C for 4 h after the addition was complete. The solution was concentrated to dryness, and ethyl acetate (25 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 20:1) to give the title compound 13 (30 mg, 14.72%).

¹ H NMR (400MHz, CDCl ₃ )δ8.08(d,1H),7.94–7.78(m,3H),7.47(m,3H),5.21(dd,1H),4.15–3.92(m,2H),3.7 6(m,1H),3.38–3.19(m,3H),2.95(dt,3H),1.87(d,2H).LC-MS(ESI):m/z＝425.1[M+H] ⁺ .

Example 14: (S)-N-((S)-1-cyano-2-(3-fluoro-4'-((4-methylpiperazin-1-yl)methyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 14)

(S)-N-((S)-1-cyano-2-(3-fluoro-4'-((4-methylpiperazin-1-yl)methyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 14)

Step 1: (S)-(1-cyano-2-(3-fluoro-4'-((4-methylpiperazin-1-yl)methyl)-[1,1'-biphenyl]-4-yl)ethyl)tert-butyl carbamate (14B)

tert-butyl-(S)-(1-cyano-2-(3-fluoro-4'-((4-methylpiperazin-1-yl)methyl)-[1,1'-biphenyl]-4-yl)ethyl)carbamate(14B)

14A (2.54 g, 8.04 mmol), INT-2 (2.3 g, 6.70 mmol), potassium carbonate (1.85 g, 13.4 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (0.98 g, 1.34 mmol) were dissolved in 1,4-dioxane (100 ml) and water (10 ml). The mixture was purged with nitrogen three times, refluxed at 95 °C for 3 h, filtered, and the filter cake was washed with ethyl acetate (100 ml). The filtrate was dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (methanol:dichloromethane (v/v) = 0%–15%) to give the title compound 14B, a dark brown solid (3.00 g, 99.0%). LC-MS (ESI): m/z = 453.3 [M+H] ⁺ .

Step 2: (S)-2-amino-3-(3-fluoro-4'-((4-methylpiperazin-1-yl)methyl)-[1,1'-biphenyl]-4-yl)propionitrile (14C)

(S)-2-amino-3-(3-fluoro-4'-((4-methylpiperazin-1-yl)methyl)-[1,1'-biphenyl]-4-yl)propanenitrile(14C)

14B (3.26 g, 7.2 mmol) was dissolved in formic acid (40 ml) and stirred overnight at room temperature. Water (40 ml) and dichloromethane (80 ml) were added, and the pH was adjusted to alkaline with sodium bicarbonate. The organic phase was separated, and the aqueous phase was extracted again with dichloromethane (80 ml x 2). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (methanol:dichloromethane (v/v) = 0%–10%) to give the title compound 14C, a dark brown oil (2.32 g, 91.3%). LC-MS (ESI): m/z = 353.3 [M+H] ⁺ .

Step 3: (S)-2-(((S)-1-cyano-2-(3-fluoro-4'-((4-methylpiperazin-1-yl)methyl)-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (14D)

tert-butyl-(S)-2-(((S)-1-cyano-2-(3-fluoro-4'-((4-methylpiperazin-1-yl)methyl)-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(14D)

14C (500 mg, 1.42 mmol), INT-3 (348 mg, 1.42 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (648 mg, 1.70 mmol), and DIPEA (366 mg, 2.84 mmol) were dissolved in dichloromethane (15 mL). The mixture was stirred at room temperature for 4 h. The reaction solution was washed with saturated sodium bicarbonate solution (10 mL x 3), the organic phase was separated, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (methanol:dichloromethane (v/v) = 0%–10%) to give the title compound 14D, a yellow solid (400 mg, 48.6%). LC-MS (ESI): m/z = 580.3 [M+H] ⁺ .

Step 4: (S)-N-((S)-1-cyano-2-(3-fluoro-4'-((4-methylpiperazin-1-yl)methyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 14)

(S)-N-((S)-1-cyano-2-(3-fluoro-4'-((4-methylpiperazin-1-yl)methyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 14)

14D (400 mg, 0.69 mmol) and 2,6-dimethylpyridine (74 mg, 0.69 mmol) were dissolved in dichloromethane (20 mL) and stirred in an ice-water bath. Tert-butyldimethylsilyltrifluoromethanesulfonate (550 mg, 2.07 mmol) was added dropwise to the reaction flask. After the addition was complete, the ice-water bath was removed, and the reaction was stirred at room temperature for 2 h. The reaction solution was washed with saturated sodium bicarbonate solution (20 mL x 2), the organic phase was separated, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (methanol:dichloromethane (v/v) = 0%–15%) to give title compound 14 (57 mg, 17.2%). LC-MS (ESI): m/z = 480.3 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.69(d,1H),7.64(d,2H),7.54–7.46(m,2H),7.43(t,1H),7.37(d,2H),5.04(dd,1H),4.01(dd,1H),3.90–3.82(m,1H),3.73(ddd,1H) ,3.48(s,2H),3.19(dd,1H),3.06(dd,1H),2.83–2.74(m,1H),2.68–2.61(m,1H),2.58(dd,1H),2.35(d,8H),2.15(s,3H),1.74(ddd,2H).

Example 15: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(4-methylthiazo-2-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 15)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(4-methylthiazol-2-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 15)

Step 1: (S)-(1-cyano-2-(2-fluoro-4-(4-methylthiazolyl-2-yl)phenyl)ethyl)tert-butyl carbamate (15B)

tert-butyl(S)-(1-cyano-2-(2-fluoro-4-(4-methylthiazol-2-yl)phenyl)ethyl)carbamate(15B)

15A (0.23 g, 1.28 mmol), INT-2 (0.50 g, 1.28 mmol), 1,1'-(diphenylphosphine)ferrocene]dichloropalladium (94 mg, 0.13 mmol), and potassium carbonate (0.53 g, 3.84 mmol) were dissolved in a mixed solvent of dioxane (27 mL) and water (3 mL). The reaction was carried out at 90 °C for 5 h under nitrogen protection. After cooling to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic layers were combined and washed successively with saturated sodium bicarbonate (20 mL) and saturated brine (20 mL). The mixture was dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (PE:EA (v/v) = 2:1) to give the title compound 15B (0.25 g, 53%). LC-MS (ESI): m/z = 362.1 [M+H] ⁺ .

Step 2: (S)-2-amino-3-(2-fluoro-4-(4-methylthiazolyl-2-yl)phenyl)propionitrile (15C)

(S)-2-amino-3-(2-fluoro-4-(4-methylthiazol-2-yl)phenyl)propanenitrile(15C)

15B (0.25 g, 0.68 mmol) was dissolved in anhydrous formic acid (4 mL) and reacted at 50 °C for 20 min. After cooling to room temperature, most of the solvent was removed by concentration. A saturated sodium bicarbonate solution (20 mL) was added to the residue, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 97:3) to give the title compound 15C (0.17 g, 96%). LC-MS (ESI): m/z = 262.2 [M+H] ⁺ .

Step 3: (S)-2-(((S)-1-cyano-2-(2-fluoro-4-(4-methylthiazo-2-yl)phenyl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (15D)

tert-butyl(S)-2-(((S)-1-cyano-2-(2-fluoro-4-(4-methylthiazol-2-yl)phenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(15D)

INT-3 (0.18 g, 0.73 mmol) was dissolved in DMF (5 mL). Under nitrogen protection, HATU (0.42 g, 1.09 mmol) and DIPEA (0.28 g, 2.19 mmol) were added. After stirring at room temperature for 20 min, 15C (0.19 g, 0.73 mmol) was added, and the reaction was allowed to proceed for 1 h at room temperature. 30 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (15 mL x 5). The organic layers were combined, dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 97:3) to give the title compound 15D (0.26 g, 73%). LC-MS (ESI): m/z = 489.1 [M+H] ⁺ .

Step 4: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(4-methylthiazo-2-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 15)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(4-methylthiazol-2-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 15)

15D (0.26 g, 0.53 mmol) was dissolved in formic acid (4.0 mL), and the mixture was reacted at 50 °C for 30 min after the addition was complete. The solution was concentrated to dryness, and ethyl acetate (25 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 1:2) to give the title compound 15 (80 mg, 39%).

^1H NMR (400MHz, DMSO-d6 ₎ )δ8.97(d,J＝8.4Hz,1H),7.74–7.66(m,2H),7.48(t,J＝7.8Hz,1H),7.39(s,1H), 5.05(m,1H),4.31(d,J＝9.4Hz,1H),3.90(m,1H),3.76(m,1H),3.36–3.32(m,1H) ,3.21(m,1H),3.15–3.07(m,1H),2.97(m,1H),2.82(dd,J＝14.0,9.4Hz,1H),2.4 3(s,3H),1.92(p,J＝5.6Hz,2H),1.29–1.21(m,1H).LC-MS(ESI):m/z＝389.1[M+H] ⁺ .

Example 16: (S)-N-((S)-2-(4-(1-acetylindoline-5-yl)-2-fluorophenyl)-1-cyanoethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 16)

(S)-N-((S)-2-(4-(1-acetylindolin-5-yl)-2-fluorophenyl)-1-cyanoethyl)-1,4-oxazepane-2-carboxamide(compound 16)

Step 1: (S)-(2-(4-(1-acetylindoline-5-yl)-2-fluorophenyl)-1-cyanoethyl)tert-butyl carbamate 16B)

tert-butyl(S)-(2-(4-(1-acetylindolin-5-yl)-2-fluorophenyl)-1-cyanoethyl)carbamate(16B)

16A (0.200 g, 0.83 mmol), INT-2 (0.323 g, 0.83 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (0.067 g, 0.083 mmol), and potassium carbonate (0.343 g, 2.49 mmol) were dissolved in 1,4-dioxane (10 mL), water (2 mL) was added, and the mixture was purged three times with nitrogen. The reaction was carried out at 90°C for 4 hours under a nitrogen atmosphere. The reaction solution was concentrated to dryness, dissolved in dichloromethane, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 5:1) to give the title compound 16B, a white solid (0.100 g, 28.4%). LC-MS (ESI): m/z = 424.1 [M+H] ⁺ .

Step 2: (S)-3-(4-(1-acetylindoline-5-yl)-2-fluorophenyl)-2-aminopropionitrile 4-methylbenzenesulfonate (16C)

(S)-3-(4-(1-acetylindolin-5-yl)-2-fluorophenyl)-2-aminopropanenitrile4-methylbenzenesulfonate(16C)

16B (0.100 g, 0.24 mmol) was dissolved in acetonitrile (2 mL), and p-toluenesulfonic acid monohydrate (0.134 g, 0.71 mmol) was added. After the addition was complete, the mixture was reacted at room temperature for 16 h. The mixture was filtered, and the filter cake was washed once with acetonitrile (1 mL). The filter cake was then evaporated to dryness to give the title compound 16C, a white solid (0.100 g, 85.5%), which was used directly in the next step of the reaction.

Step 3: (S)-2-(((S)-2-(4-(1-acetylindoline-5-yl)-2-fluorophenyl)-1-cyanoethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (16D)

tert-butyl tert-butyl(S)-2-(((S)-2-(4-(1-acetylindolin-5-yl)-2-fluorophenyl)-1-cyanoethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(16D)

INT-3 (0.074 g, 0.303 mmol) was dissolved in dichloromethane (5 mL), followed by the addition of triethylamine (0.061 g, 0.606 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate (0.115 g, 0.303 mmol). After the addition was complete, the mixture was stirred at room temperature for 1 hour. Then, 16C (0.100 g, 0.202 mmol) was added, and the mixture was allowed to react overnight at room temperature. After the reaction was complete, the mixture was concentrated, and the crude 16D was used directly in the next step. LC-MS (ESI): m/z = 549.2 [MH] ^- .

Step 4: (S)-N-((S)-2-(4-(1-acetylindoline-5-yl)-2-fluorophenyl)-1-cyanoethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 16)

(S)-N-((S)-2-(4-(1-acetylindolin-5-yl)-2-fluorophenyl)-1-cyanoethyl)-1,4-oxazepane-2-carboxamide(compound 16)

The crude compound 16D was dissolved in acetonitrile (10 mL) and p-toluenesulfonic acid monohydrate (0.134 g, 0.71 mmol) was added. After the addition was complete, the mixture was reacted at room temperature for 16 h. The mixture was concentrated to dryness, and ethyl acetate (25 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 20:1) to give the title compound 16 (27 mg, two-step yield 29.7%).

¹ H NMR (400MHz, CD ₃ OD)δ8.15(d,1H),7.51(s,1H),7.47–7.28(m,4H),5.14(dd,1H),4.23–4.09(m,3H),4.05–3.96(m,1H),3.82-3.74(m,1H),3.34(s,1H) ,3.30–3.17(m,5H),3.05–2.95(m,1H),2.93–2.82(m,1H),2.72(dd,1H),2.25(s,3H),1.97–1.83(m,2H).LC-MS(ESI):m/z＝451.2[M+H] ⁺ .

Example 17: (S)-N-((S)-1-cyano-2-(4'-cyano-3'-cyclopropyl-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 17)

(S)-N-((S)-1-cyano-2-(4'-cyano-3'-cyclopropyl-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 17)

Step 1: (S)-(1-cyano-2-(4'-cyano-3'-cyclopropyl-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)tert-butyl carbamate (17B)

tert-butyl(S)-(1-cyano-2-(4'-cyano-3'-cyclopropyl-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)carbamate(17B)

17A (0.300 g, 1.67 mmol), INT-2 (0.646 g, 1.67 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (0.134 g, 0.167 mmol), and potassium carbonate (0.686 g, 4.98 mmol) were dissolved in 1,4-dioxane (15 mL), water (3 mL) was added, and the mixture was purged three times with nitrogen. The reaction was carried out at 100°C for 16 hours under a nitrogen atmosphere. The reaction solution was concentrated to dryness, dissolved in dichloromethane, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 5:1) to give the title compound 17B, a white solid (0.380 g, 55.4%). LC-MS (ESI): m/z = 406.1 [M+H] ⁺ .

Step 2: (S)-4'-(2-amino-2-cyanoethyl)-3-cyclopropyl-3'-fluoro-[1,1'-biphenyl]-4-nitrile 4-methylbenzenesulfonate

(S)-4'-(2-amino-2-cyanoethyl)-3-cyclopropyl-3'-fluoro-[1,1'-biphenyl]-4-carbonitrile 4-methylbenzenesulfonate(17C)

17B (0.380 g, 0.94 mmol) was dissolved in acetonitrile (5 mL), and p-toluenesulfonic acid monohydrate (534 g, 2.81 mmol) was added. After the addition was complete, the mixture was reacted at room temperature for 16 h. The mixture was filtered, and the filter cake was washed once with acetonitrile (2 mL). The filter cake was then evaporated to dryness to give the title compound 17C, a white solid (0.360 g, 80.5%), which was used directly in the next step of the reaction.

Step 3: (S)-2-(((S)-1-cyano-2-(4'-cyano-3'-cyclopropyl-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazacycloheptane-4-carboxylic acid tert-butyl ester (17D)

tert-butyl(S)-2-(((S)-1-cyano-2-(4'-cyano-3'-cyclopropyl-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(17D)

INT-3 (0.123 g, 0.503 mmol) was dissolved in dichloromethane (5 mL), followed by the addition of triethylamine (0.101 g, 1.00 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate (0.193 g, 0.503 mmol). The mixture was stirred at room temperature for 1 hour after the addition was complete, then 17C (0.160 g, 0.335 mmol) was added, and the mixture was allowed to react overnight at room temperature. The reaction mixture was then concentrated, and the crude 17D was used directly in the next reaction. LC-MS (ESI): m/z = 531.2 [MH] ^- .

Step 4: (S)-N-((S)-1-cyano-2-(4'-cyano-3'-cyclopropyl-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 17)

(S)-N-((S)-1-cyano-2-(4'-cyano-3'-cyclopropyl-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 17)

The crude compound 17D was dissolved in acetonitrile (10 mL) and p-toluenesulfonic acid monohydrate (0.190 g, 1.00 mmol) was added. After the addition was complete, the mixture was reacted at room temperature for 16 h. The mixture was concentrated to dryness, and ethyl acetate (25 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 20:1) to give the title compound 17 (60 mg, two-step yield 41.4%).

^1H NMR (400MHz, CDCl3 ₎ )δ7.66(d,1H),7.44–7.37(m,2H),7.33(dd,1H),7.28(d,2H),7.10(s,1H), 5.15(dt,1H),4.22(dt,1H),4.12–3.99(m,1H),3.85–3.73(m,1H),3.57–3. 41(m,1H),3.30–3.17(m,2H),3.13–2.97(m,3H),2.34(ddd,1H),2.07–1.94 (m,2H),1.24–1.15(m,2H),0.92–0.83(m,2H).LC-MS(ESI):m/z＝433.2[M+H] ⁺ .

Example 18: (S)-N-((S)-1-cyano-2-(3-fluoro-4'-(methylcarbamoyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 18)

(S)-N-((S)-1-cyano-2-(3-fluoro-4'-(methylcarbamoyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 18)

Step 1: (S)-(1-cyano-2-(3-fluoro-4'-(methylcarbamoyl)-[1,1'-biphenyl]-4-yl)ethyl)tert-butyl carbamate (18B)

tert-butyl(S)-(1-cyano-2-(3-fluoro-4'-(methylcarbamoyl)-[1,1'-biphenyl]-4-yl)ethyl)carbamate(18B)

Compound 18A (0.50 g, 1.46 mmol), 2a (0.26 g, 1.46 mmol), 1,1'-(diphenylphosphine)ferrocene]dichloropalladium (0.11 mg, 0.15 mmol), and potassium carbonate (0.61 g, 4.38 mmol) were dissolved in a mixed solvent of dioxane (27 mL) and water (3 mL). The reaction was carried out at 90 °C for 5 h under nitrogen protection. After cooling to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic layers were combined and washed successively with saturated sodium bicarbonate (20 mL) and saturated brine (20 mL). The mixture was dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (PE:EA (v/v) = 2:1) to give the title compound 18B (0.34 g, 59%). LC-MS (ESI): m/z = 398.2 [M+H] ⁺ .

Step 2: (S)-4'-(2-amino-2-cyanoethyl)-3'-fluoro-N-methyl-[1,1'-biphenyl]-4-carboxamide (18C)

(S)-4'-(2-amino-2-cyanoethyl)-3'-fluoro-N-methyl-[1,1'-biphenyl]-4-carboxamide(18C)

18B (0.34 g, 0.86 mmol) was dissolved in anhydrous formic acid (4 mL) and reacted at 50 °C for 20 min. After cooling to room temperature, most of the solvent was removed by concentration. A saturated sodium bicarbonate solution (20 mL) was added to the residue, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 97:3) to give the title compound 18C (0.22 g, 87%). LC-MS (ESI): m/z = 298.1 [M+H] ⁺ .

Step 3: (S)-2-(((S)-1-cyano-2-(3-fluoro-4'-(methylcarbamoyl)-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (18D)

tert-butyl(S)-2-(((S)-1-cyano-2-(3-fluoro-4'-(methylcarbamoyl)-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(18D)

INT-3 (0.23 g, 0.76 mmol) was dissolved in DMF (5 mL). Under nitrogen protection, HATU (0.43 g, 1.14 mmol) and DIPEA (0.29 g, 2.28 mmol) were added. After stirring at room temperature for 20 min, 18C (0.23 g, 0.76 mmol) was added, and the reaction was allowed to proceed for 1 h at room temperature. 30 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (15 mL x 5). The organic layers were combined, dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 97:3) to give the title compound 18D (0.31 g, 77%). LC-MS (ESI): m/z = 469.2 [M-56+H] ⁺ .

Step 4: (S)-N-((S)-1-cyano-2-(3-fluoro-4'-(methylcarbamoyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 18)

(S)-N-((S)-1-cyano-2-(3-fluoro-4'-(methylcarbamoyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 18)

18D (0.31 g, 0.59 mmol) was dissolved in formic acid (4.0 mL), and the mixture was reacted at 50 °C for 30 min after the addition was complete. The solution was concentrated to dryness, and ethyl acetate (25 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 1:2) to give the title compound 18 (50 mg, 20%).

¹ H NMR (400MHz, DMSO-d ₆ )δ8.98(d,1H),8.49(d,1H),7.93(d,2H),7.81(d,2H),7.64–7.57(m,2H),7.47(t,1H),5.06(m,1H),4.35–4.29(m,1H),3.91(m,1H),3 .77(m,1H),3.38(m,1H),3.18(m,1H),2.98(m,2H),2.87(m,1H),2.80(d,3H),1.92(m,2H),1.23(m,1H).LC-MS(ESI):m/z＝425.2[M+H] ⁺ .

Example 19: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-methyl-1H-indol-2-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 19)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-methyl-1H-indol-2-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

Step 1: (S)-(1-cyano-2-(2-fluoro-4-(1-methyl-1H-indol-2-yl)phenyl)ethyl)tert-butyl carbamate (19B)

tert-butyl(S)-(1-cyano-2-(2-fluoro-4-(1-methyl-1H-indol-2-yl)phenyl)ethyl)carbamate

2a (0.40 g, 1.17 mmol) was dissolved in 1,4-dioxane (10 mL) and water (0.4 mL). Then, 1-methyl-2-indoleboronic acid pinacol ester (0.39 g, 1.52 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloromethane dichloride complex (0.19 g, 0.23 mmol), and potassium carbonate (0.32 g, 2.34 mmol) were added. Under nitrogen protection, the mixture was heated to 90 °C for 2 h. The reaction was allowed to proceed overnight at room temperature. The mixture was concentrated to dryness, and saturated ammonium chloride aqueous solution (50 mL) was added. Extraction was performed with ethyl acetate (20 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 10:1, 4:1) to give title compound 19B, a white solid (0.25 g, 54.31%). LC-MS (ESI): m/z=394.2[M+H] ⁺ .

Step 2: (S)-2-amino-3-(2-fluoro-4-(1-methyl-1H-indol-2-yl)phenyl)propionitrile (19C)

(S)-2-amino-3-(2-fluoro-4-(1-methyl-1H-indol-2-yl)phenyl)propanenitrile

19B (0.37 g, 0.94 mmol) was dissolved in acetonitrile (30 mL), and p-toluenesulfonic acid monohydrate (0.54 g, 2.82 mmol) was added. After the addition was complete, the mixture was reacted at room temperature for 4 h. The solution was concentrated to dryness, and ethyl acetate (25 mL) was added. The pH was adjusted to approximately 8 by dropwise addition of saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 1:1) to give title 19C, a pale yellow solid (0.25 g, 90.67%). LC-MS (ESI): m/z = 294.3 [M+H] ⁺ .

Step 3: (S)-2-(((S)-1-cyano-2-(2-fluoro-4-(1-methyl-1H-indol-2-yl)phenyl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (19D)

tert-butyl(S)-2-(((S)-1-cyano-2-(2-fluoro-4-(1-methyl-1H-indol-2-yl)phenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate

19C (0.25 g, 0.85 mmol) was dissolved in dichloromethane (10 mL), and INT-3 (0.21 g, 0.85 mmol), diisopropylethylamine (0.55 g, 4.25 mmol), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (0.32 g, 0.85 mmol) were added. The mixture was reacted overnight at room temperature. A saturated sodium chloride aqueous solution (30 mL) was added, and the mixture was extracted with ethyl acetate (25 mL). The organic phase was washed with saturated sodium chloride aqueous solution (25 mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 2:1) to give title compound 19D, a white solid (0.40 g, 90.39%). LC-MS (ESI): m/z = 485.1 [M-57+H] ⁺ .

Step 4: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-methyl-1H-indol-2-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 19)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-methyl-1H-indol-2-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

19D (0.40 g, 0.77 mmol) was dissolved in acetonitrile (30 mL), and p-toluenesulfonic acid monohydrate (0.44 g, 2.31 mmol) was added. After the addition was complete, the mixture was reacted at room temperature for 4 h. The solution was concentrated to dryness, and ethyl acetate (25 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 20:1) to give title compound 19, a pale yellow solid (0.15 g, 46.33%).

¹ H NMR (400MHz, CDCl ₃ )δ7.64(d,1H),7.46–7.03(m,6H),6.59(s,1H),5.25–5.09(m,1H),4.15(dd,1H),4.07–3.90(m,1H),3.82–3 .61(m,4H),3.37(dd,1H),3.25(t,2H),3.12–2.86(m,3H),1.93–1.75(m,2H).LC-MS(ESI):m/z＝421.2[M+H] ⁺ .

Example 20: (S)-N-((S)-1-cyano-2-(4'-(dimethylphosphoryl)-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 20)

(S)-N-((S)-1-cyano-2-(4'-(dimethylphosphoryl)-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 20)

Step 1: (S)-(1-cyano-2-(4'-(dimethylphosphoryl)-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)tert-butyl carbamate (20B)

tert-butyl(S)-(1-cyano-2-(4'-(dimethylphosphoryl)-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)carbamate(20B)

Compound 20A (0.200 g, 1.29 mmol), INT-2 (0.502 g, 1.29 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (0.105 g, 0.129 mmol), and potassium carbonate (0.534 g, 3.87 mmol) were dissolved in 1,4-dioxane (15 mL), water (3 mL) was added, and the mixture was purged three times with nitrogen. The reaction was carried out at 90°C for 4 hours under a nitrogen atmosphere. The reaction solution was concentrated to dryness, dissolved in dichloromethane, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 5:1) to give the title compound 20B, a white solid (0.420 g, 78.5%). LC-MS (ESI): m/z = 417.2 [M+H] ⁺ .

Step 2: (S)-2-amino-3-(4'-(dimethylphosphoryl)-3-fluoro-[1,1'-biphenyl]-4-yl)propionitrile 4-methylbenzenesulfonate (20C)

(S)-2-amino-3-(4'-(dimethylphosphoryl)-3-fluoro-[1,1'-biphenyl]-4-yl)propanenitrile 4-methylbenzenesulfonate(20C)

20B (0.420 g, 1.01 mmol) was dissolved in acetonitrile (8 mL), and p-toluenesulfonic acid monohydrate (0.575 g, 3.03 mmol) was added. After the addition was complete, the mixture was reacted at room temperature for 16 h. The mixture was filtered, and the filter cake was rinsed once with acetonitrile (2 mL). The filter cake was then evaporated to dryness to give the title compound 20C, a white solid (0.500 g, 99.9%), which was used directly in the next step of the reaction.

Step 3: (S)-2-(((S)-1-cyano-2-(4'-(dimethylphosphoryl)-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (20D)

tert-butyl(S)-2-(((S)-1-cyano-2-(4'-(dimethylphosphoryl)-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(20D)

INT-3 (0.148 g, 0.605 mmol) was dissolved in dichloromethane (10 mL), followed by the addition of triethylamine (0.124 g, 1.23 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate (0.230 g, 0.605 mmol). The reaction mixture was stirred at room temperature for 1 hour, followed by the addition of 20°C (0.200 g, 0.41 mmol) and incubation at room temperature overnight. The reaction mixture was then concentrated, and the crude product 20D was used directly in the next reaction. LC-MS (ESI): m/z = 542.2 [MH] ^- .

Step 4: (S)-N-((S)-1-cyano-2-(4'-(dimethylphosphoryl)-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 20)

(S)-N-((S)-1-cyano-2-(4'-(dimethylphosphoryl)-3-fluoro-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 20)

The crude compound 20D was dissolved in acetonitrile (10 mL), and p-toluenesulfonic acid monohydrate (0.234 g, 1.23 mmol) was added. After the addition was complete, the mixture was reacted at room temperature for 16 h, concentrated to dryness, and ethyl acetate (25 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 20:1) to give the title compound 20 (60 mg, two-step yield 33.1%).

¹ H NMR (400MHz, CDCl ₃ )δ7.83(d,1H),7.81(d,1H),7.69(dd,2H),7.41(d,2H),7.38–7.30(m,2H),5.18(dd,1H),4.20(dd,1H),4.09–3.98(m,1H),3.84–3.74(m,1H ),3.41(dd,1H),3.31–3.17(m,2H),3.06–2.91(m,3H),1.96(s,1H),1.78(d,6H),1.38(dd,1H),1.22(dd,1H).LC-MS(ESI):m/z＝444.3[M+H] ⁺ .

Example 21: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 21)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 21)

Step 1: (S)-(1-cyano-2-(2-fluoro-4-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)phenyl)ethyl)tert-butyl carbamate (21B)

tert-butyl(S)-(1-cyano-2-(2-fluoro-4-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)phenyl)ethyl)carbamate(21B)

Reactant 21A (0.47 g 2.0 mmol) and INT-2 (0.69 g 2.0 mmol) were dissolved in 1,4-dioxane (50 ml), followed by the addition of potassium carbonate (0.83 g 6.0 mmol) and Pd(dppf) _Cl₂ (0.15 g 0.2 mmol). The mixture was purged with nitrogen and reacted at 100°C. TLC and LC-MS showed a small amount of the starting material remaining. The solution was concentrated and evaporated to dryness, dissolved in DCM, filtered through diatomaceous earth, and the filtrate was evaporated to dryness and passed through a column (EA/PE = 0%–40%) to give the title compound 21B as a pale yellow solid (0.50 g, 67.3%). LC-MS (ESI): m/z = 372.3 [M+H] ^⁺ .

Step 2: (S)-2-amino-3-(2-fluoro-4-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)phenyl)propionitrile (21C)

(S)-2-amino-3-(2-fluoro-4-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)phenyl)propanenitrile(21C)

21B (0.5 g, 1.35 mmol) was dissolved in anhydrous formic acid (10 mL). The reaction was carried out at 50 °C for 0.5 h. TLC analysis showed complete substrate reaction. The formic acid was removed at low temperature, and the pH was adjusted to 8-10 with saturated sodium bicarbonate solution. Extraction was performed using DCM (20 mL x 2), and the solution was dried over anhydrous sodium sulfate. The extract was then purified by column chromatography (DCM: _CH₃OH = 10:1) to give the title compound 21C as a pale yellow liquid (0.35 g, 95.6%). LC-MS (ESI): m/z = 272.3 [M+H] ^⁺ .

Step 3: (S)-(((S)-1-cyano-2-(2-fluoro-4-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)phenyl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (21D)

tert-butyl(S)-2-(((S)-1-cyano-2-(2-fluoro-4-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)phenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(21D)

INT-3 (0.32 g, 1.29 mmol), HATU (0.59 g, 1.55 mmol), and DIPEA (0.5 mL) were dissolved in dichloromethane (30 mL) at room temperature. After stirring for 10 min at room temperature, 21C (0.35 g, 1.29 mmol) was added, and the mixture was stirred for 1 h at room temperature. The reaction was monitored by TLC until complete. The reaction was quenched with water (10 mL), and the mixture was extracted with dichloromethane (15 mL × 2). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (PE:EA = 1:1) to give the title compound 21D (0.4 g, 62%). LC-MS (ESI): m/z = 443.1 [M-56+H] ⁺ .

Step 4: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 21)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 21)

21D (0.4 g, 0.8 mmol) was dissolved in anhydrous formic acid (10 ml). After stirring at 50 °C for 0.5 h, TLC analysis showed that the substrate reaction was complete. The formic acid was removed at low temperature, and the pH was adjusted to 8-10 by adding saturated sodium bicarbonate solution. The mixture was extracted with dichloromethane (20 ml x 2), dried over anhydrous sodium sulfate, and purified by column chromatography (DCM: _CH3OH = 10:1) to give compound 21 (0.2 g, 62.7%). LC-MS (ESI): m/z = 399.1 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.71-8.69(m,1H),8.22(s,1H),7.87-7.84(m,1H),7.48-7.37(m,3H),6.48-6.46(m,1H),5.03-4.99(m,1H),4.05-4.02(m,1H) ,3.90-3.84(m,1H),3.76-3.70(m,1H),3.50(s,3H),3.25-3.09(m,4H),2.86-2.80(m,1H),2.70-2.58(m,2H),1.80-1.71(m,2H),.

Example 22: (S)-N-((S)-1-cyano-2-(4-(1-ethyl-1H-pyrazol-4-yl)-2-fluorophenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 22)

(S)-N-((S)-1-cyano-2-(4-(1-ethyl-1H-pyrazol-4-yl)-2-fluorophenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 22)

Step 1: (S)-(1-cyano-2-(4-(1-ethyl-1H-pyrazole-4-yl)-2-fluorophenyl)ethyl)tert-butyl carbamate (22B)

(S)-tert-butyl(1-cyano-2-(4-(1-ethyl-1H-pyrazol-4-yl)-2-fluorophenyl)ethyl)carbamate(22B)

2a (0.32 g, 1.46 mmol) was dissolved in DMF (10 mL), and intermediate 1 (0.5 g, 1.46 mmol), potassium carbonate (0.61 g, 4.38 mmol), and 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride (0.11 g, 0.15 mmol) were added. After the addition was complete, the mixture was reacted at 100 °C for 3 hours. The mixture was cooled to room temperature, and an aqueous solution (20 mL) was added. The mixture was extracted with dichloromethane (20 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 2:1) to give the title compound 22B, a colorless liquid (0.44 g, 84.1%). LC-MS (ESI): m/z = 359.1 [M+H] ⁺ .

Step 2: (S)-2-amino-3-(4-(1-ethyl-1H-pyrazol-4-yl)-2-fluorophenyl)propionitrile (22C)

(S)-2-amino-3-(4-(1-ethyl-1H-pyrazol-4-yl)-2-fluorophenyl)propanenitrile(22C)

22B (0.40 g, 1.12 mmol) was dissolved in formic acid (10 mL), and the mixture was reacted at 50 °C for 2 h after the addition was complete. The solution was concentrated to dryness, and ethyl acetate (25 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 22C, a colorless liquid (0.29 g, 100%), which was used directly in the next reaction.

Step 3: (S)-2-(((S)-1-cyano-2-(4-(1-ethyl-1H-pyrazole-4-yl)-2-fluorophenyl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (22D)

(S)-tert-butyl 2-(((S)-1-cyano-2-(4-(1-ethyl-1H-pyrazol-4-yl)-2-fluorophenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(22D)

22C (0.30 g, 1.16 mmol) was dissolved in DMF (10 mL), and INT-3 (0.28 g, 1.16 mmol), diisopropylethylamine (0.45 g, 3.48 mmol), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (0.66 g, 1.74 mmol) were added. The reaction was allowed to proceed overnight at room temperature. A saturated sodium chloride aqueous solution (30 mL) was added, and the mixture was extracted with ethyl acetate (25 mL). The organic phase was washed with saturated sodium chloride aqueous solution (25 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 22D, a pale yellow solid (0.30 g, 53.3%), which was used directly in the next reaction. LC-MS (ESI): m/z = 430.1 [M-57+H] ⁺ .

Step 4: (S)-N-((S)-1-cyano-2-(4-(1-ethyl-1H-pyrazol-4-yl)-2-fluorophenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 22)

(S)-N-((S)-1-cyano-2-(4-(1-ethyl-1H-pyrazol-4-yl)-2-fluorophenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound 22)

22D (0.15 g, 0.31 mmol) was dissolved in formic acid (2.0 mL), and the mixture was reacted at 35 °C for 4 h after the addition was complete. The solution was concentrated to dryness, and ethyl acetate (25 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 1:2) to give the title compound 22 (80 mg, 66.9%).

¹ H NMR (400MHz, CDCl ₃ )δ9.43(s,1H),9.10–9.08(m,1H),8.27(s,1H),7.92(s,1H),7.44–7.31 (m,3H),5.03–4.96(m,1H),4.51–4.48(m,1H),4.17–4.11(m,2H),3.96–3 .90(m,1H),5.82–4.76(m,1H),3.47–3.43(m,1H),3.29–3.21(m,2H),3.1 6–3.07(m,2H),2.98–2.92(m,1H),2.12–1.96(m,2H),1.42–1.38(m,3H). LC-MS (ESI): m/z=386.1[M+H] ⁺ .

Example 23: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(3-(methyl-d3)-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 23)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(3-(methyl-d3)-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound23)

Step 1: 5-Chloro-3-(methyl-d3)benzo[d]oxazol-2(3H)-one (23B)

5-chloro-3-(methyl-d3)benzo[d]oxazol-2(3H)-one(23B)

23A (5 g, 29.49 mmol) was dissolved in DMF (25 mL), and cesium carbonate (10.57 g, 32.44 mmol) and deuterated iodomethane (4.28 g, 29.52 mmol) were added. The reaction was carried out at 25 °C for 4 hours. After the reaction was completed, the reaction solution was poured into water (60 mL), filtered, and the filter cake was collected. The filter cake was dissolved in dichloromethane, washed with saturated brine, and the organic phase was collected. The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 23B as a brown solid (5.4 g, yield 98.1%). LCMS m/z = 187.0 [M+1] ⁺

Step 2: 3-(methyl-d3)-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)benzo[d]oxazol-2(3H)-one (23C)

3-(methyl-d3)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]oxazol-2(3H)-one(23C)

Compound 23B (5.4 g, 28.94 mmol) was dissolved in 1,4-dioxane (54 mL), and pinacol borane (12.49 g, 49.20 mmol), potassium acetate (8.52 g, 86.82 mmol), palladium acetate (652 mg, 2.89 mmol), and X-Phos (2.73 g, 5.79 mmol) were added. After the addition was complete, the mixture was heated to 100 °C and reacted for 3 hours. After the reaction was complete, the mixture was filtered, and the filtrate was concentrated. The residue was purified by silica gel column chromatography (eluting ratio PE:EA (v/v) = 1:0-10:1) to give 23C, a white solid (7.2 g, yield 89.8%). LC-MS m/z = 279.2 [M+1] ⁺

Step 3: (S)-(1-cyano-2-(2-fluoro-4-(3-(methyl-d3)-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)tert-butyl carbamate (23D)

tert-butyl(S)-(1-cyano-2-(2-fluoro-4-(3-(methyl-d3)-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)carbamate(23D)

Compound 23C (2.0 g, 7.19 mmol) was dissolved in a mixed solvent of 1,4-dioxane (20 ml) and water (4 ml). Compound 2a (2.47 mg, 7.19 mmol) was added, followed by potassium carbonate (2.98 g, 21.57 mmol), palladium acetate (161 mg, 0.72 mmol), and X-Phos (686 mg, 1.44 mmol). After three nitrogen purgings, the mixture was heated to 100 °C and reacted for 5 hours. The reaction was then concentrated, and the residue was purified by silica gel column chromatography (eluting ratio PE:EA (v/v) = 1:0–5:1) to give compound 23D, a white solid (2.57 g, 86.2%). LCMS m/z = 415.3 [M+1] ⁺

Step 4: (S)-2-amino-3-(2-fluoro-4-(3-(methyl-d3)-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)propionitrile (23E)

(S)-2-amino-3-(2-fluoro-4-(3-(methyl-d3)-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)propanenitrile(23E)

Compound 23D (1 g, 2.41 mmol) was dissolved in anhydrous formic acid (10 mL) and stirred at 25 °C for 5 hours. After the reaction was complete, the reaction solution was slowly poured into a saturated sodium bicarbonate solution (100 mL), pH 7–8, and then extracted with dichloromethane (50 mL × 3). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 23E, a pale yellow solid (710 mg, 93.5%). LCMS m/z = 315.2 [M+1] ⁺

Step 5: (S)-2-(((S)-1-cyano-2-(2-fluoro-4-(3-(methyl-d3)-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazacycloheptane-4-carboxylic acid tert-butyl ester (23F)

tert-butyl(S)-2-(((S)-1-cyano-2-(2-fluoro-4-(3-(methyl-d3)-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(23F)

Under nitrogen protection, compound 23E (656 mg, 2.07 mmol) was dissolved in DMF (7 mL), and INT-3 (510 mg, 2.07 mmol), diisopropylethylamine (1.34 g, 10.35 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.18 g, 3.10 mmol) were added. After the addition was complete, the mixture was heated to 25 °C and reacted for 3 hours. After the reaction was complete, the reaction solution was poured into water (20 mL) and extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with saturated brine (40 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by silica gel column chromatography (eluent ratio PE:EA (v/v) = 1:0 to 1:1) to give compound 23F, a yellow solid (837 mg, 74.7%). LCMS m/z = 486.3[M-tBu+1] ⁺

Step 6: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(3-(methyl-d3)-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 23)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(3-(methyl-d3)-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide(compound23)

Compound 23F (837 mg, 1.55 mmol) was dissolved in anhydrous formic acid (3 mL) and stirred at 25 °C for 3 hours. After the reaction was complete, the reaction solution was slowly poured into a saturated sodium bicarbonate solution (30 mL), pH 7–8, and then extracted with dichloromethane (50 mL × 3). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by preparative liquid chromatography (HPLC). HPLC conditions: C18 reversed-phase column; mobile phase: deionized water (A) containing 0.1% trifluoroacetic acid; acetonitrile (B) containing 0.1% trifluoroacetic acid; gradient elution; mobile phase B concentration: 5%–50%; elution time: 15 min; flow rate: 12 mL/min; column temperature: 30 °C; retention time: 3.7 min. Compound 23 (300 mg, yield 43.8%) was obtained.

¹ H NMR (400MHz, DMSO-d ₆ )δ8.76(d,1H),7.65(d,J＝1.8,1H),7.62–7.50(m,2H),7.49–7.42(m,2H),7.38(m,1H),5.05(m,1H),4.13(m,1H),3.99–3.82(m ,1H),3.74(m,1H),3.65–3.52(m,1H),3.30(m,2H),3.16(m,2H),3.00–2.80(m,1H),2.74–2.58(m,1H),1.88–1.68(m,2H).LCMS m/z＝442.1[M+1] ⁺

Example 24: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinoline-6-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 24)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

Step 1: 6-Bromo-2-methyl-3,4-dihydroisoquinoline-1(2H)-one (24B)

6-bromo-2-methyl-3,4-dihydroisoquinolin-1(2H)-one

24A (0.57 g, 2.5 mmol) was dissolved in dry N,N-dimethylformamide (20 mL), cooled to 0 °C under nitrogen protection, and sodium hydride (0.12 g, 3.0 mmol, 60% wt) was added in portions. After the addition was complete, the reaction was allowed to proceed for 20 minutes, and then iodomethane (0.51 g, 3.6 mmol) was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature for 30 minutes. The reaction was quenched with water (100 mL), extracted with ethyl acetate (100 mL × 2), the organic phases were combined, washed with saturated brine (100 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by silica gel column chromatography (PE:EA = 6:1) to give the target compound 6-bromo-2-methyl-3,4-dihydroisoquinoline-1(2H)-one (24B), a yellow solid (0.48 g, 79% yield).

Step 2: (S)-(1-cyano-2-(2-fluoro-4-(2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinoline-6-yl)phenyl)ethyl)tert-butyl carbamate (24C)

tert-butyl(S)-(1-cyano-2-(2-fluoro-4-(2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)phenyl)ethyl)carbamate

24B (0.29 g, 1.2 mmol), INT-2 (0.39 g, 1.0 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (71 mg, 0.1 mmol), and potassium carbonate (0.28 g, 2.0 mmol) were added sequentially to 1,4-dioxane (15 mL) and water (3 mL). The system was purged three times with nitrogen and reacted at 100 °C for 2 hours. After the reaction was complete, the mixture was cooled to room temperature, and water (50 mL) was added. The aqueous phase was extracted with ethyl acetate (50 mL × 2). The combined organic phases were washed with saturated sodium chloride aqueous solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by silica gel column chromatography (PE:EA = 3:1 to 2:1) to give 24C, a brown solid (0.35 g, yield 83%). LCMS m/z = 424.2 [M+1] ⁺ .

Step 3: (S)-2-amino-3-(2-fluoro-4-(2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinoline-6-yl)phenyl)propionitrile (24D)

(S)-2-amino-3-(2-fluoro-4-(2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)phenyl)propanenitrile

24C (0.35 g, 0.83 mmol) was dissolved in formic acid (6.0 mL), and the mixture was reacted at 50 °C for 10 minutes after the addition was complete. The solution was concentrated to dryness, and ethyl acetate (60 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated, and the aqueous phase was extracted with ethyl acetate (60 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 24D (0.24 g, 89% yield).

Step 4: (S)-2-(((S)-1-cyano-2-(2-fluoro-4-(2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinoline-6-yl)phenyl)ethyl)carbamoyl)-1,4-oxazacycloheptane-4-carboxylic acid tert-butyl ester (24E)

tert-butyl(S)-2-(((S)-1-cyano-2-(2-fluoro-4-(2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)phenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate

24D (0.24 g, 0.74 mmol) was dissolved in DMF (10 mL), and INT-3 (0.25 g, 1.0 mmol), diisopropylethylamine (0.19 g, 1.5 mmol), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (0.38 g, 1 mmol) were added. The mixture was reacted at room temperature for 1 hour after the addition was complete. Water (30 mL) was added, and the mixture was extracted with ethyl acetate (60 mL × 2). The organic phase was washed with saturated sodium chloride aqueous solution (60 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated by silica gel column chromatography (PE:EA = 2:1 to 1:2) to give the title compound 24E as a pale yellow solid (0.28 g, yield 69%).

Step 5: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinoline-6-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 24)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

24E (0.28 g, 0.51 mmol) was dissolved in formic acid (6.0 mL) and reacted at 50 °C for 10 min. The mixture was concentrated under reduced pressure, and ethyl acetate (60 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate solution. The organic layer was separated and extracted with ethyl acetate (60 mL × 5). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 10:1) to give compound 24 (0.18 g, yield 78%).

¹ H NMR (400MHz, DMSO-d ₆ )δ8.82(d,1H),7.93(d,1H),7.71–7.63(m,2H),7.63–7.53(m,2H),7.47 (t,1H),5.11–5.01(m,1H),4.15(dd,1H),3.94-3.84(m,1H),3.80-3.70( m,1H),3.58(t,2H),3.37–3.16(m,4H),3.08–3.01(m,4H),3.00-2.90(m, 1H),2.89–2.65(m,2H),1.88-1.74(m,2H).LC-MS(ESI):m/z＝451.3[M+H] ⁺ .

Example 25: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(2-methyl-1-oxoisoindoline-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 25)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(2-methyl-1-oxoisoindolin-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

Step 1: (S)-(1-cyano-2-(2-fluoro-4-(2-methyl-1-oxoisoindoline-5-yl)phenyl)ethyl)tert-butyl carbamate (25B)

tert-butyl(S)-(1-cyano-2-(2-fluoro-4-(2-methyl-1-oxoisoindolin-5-yl)phenyl)ethyl)carbamate

25A (0.27 g, 1.2 mmol), INT-2 (0.36 g, 0.92 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (71 mg, 0.1 mmol), and potassium carbonate (0.28 g, 2.0 mmol) were added sequentially to 1,4-dioxane (15 mL) and water (3 mL). The system was purged three times with nitrogen and reacted at 100 °C for 2 hours. After the reaction was complete, the mixture was cooled to room temperature, and water (50 mL) was added. The aqueous phase was extracted with ethyl acetate (50 mL × 2). The combined organic phases were washed with saturated sodium chloride aqueous solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by silica gel column chromatography (PE:EA = 3:1 to 2:1) to give 25B, a brown solid (0.36 g, 95% yield). LCMS m/z = 410.1 [M+1] ⁺ .

Step 2: (S)-2-amino-3-(2-fluoro-4-(2-methyl-1-oxoisoindoline-5-yl)phenyl)propionitrile (25C)

(S)-2-amino-3-(2-fluoro-4-(2-methyl-1-oxoisoindolin-5-yl)phenyl)propanenitrile

25B (0.36 g, 0.88 mmol) was dissolved in formic acid (6.0 mL), and the mixture was reacted at 50 °C for 10 minutes after the addition was complete. The solution was concentrated to dryness, and ethyl acetate (60 mL) was added. The pH was adjusted to approximately 8 by dropwise addition of saturated sodium bicarbonate solution. The organic layer was separated, and the aqueous phase was extracted with ethyl acetate (60 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 25C (0.28 g, 100% yield). LCMS m/z = 310.1 [M+1] ⁺ .

Step 3: Tert-butyl(S)-2-(((S)-1-cyano-2-(2-fluoro-4-(2-methyl-1-oxoisoindoline-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazacycloheptane-4-carboxylic acid ester (25D)

tert-butyl(S)-2-(((S)-1-cyano-2-(2-fluoro-4-(2-methyl-1-oxoisoindolin-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate

25C (0.28 g, 0.88 mmol) was dissolved in DMF (10 mL), and INT-3 (0.25 g, 1.0 mmol), diisopropylethylamine (0.19 g, 1.5 mmol), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (0.38 g, 1 mmol) were added. The mixture was reacted at room temperature for 1 hour after the addition was complete. Saturated sodium chloride aqueous solution (30 mL) was added, and the mixture was extracted with ethyl acetate (60 mL × 2). The organic phase was washed with saturated sodium chloride aqueous solution (60 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated by silica gel column chromatography (PE:EA = 2:1 to 1:2) to give the title compound 25D as a pale yellow solid (0.29 g, yield 60%).

Step 4: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(2-methyl-1-oxoisoindoline-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 25)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(2-methyl-1-oxoisoindolin-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

25D (0.29 g, 0.55 mmol) was dissolved in formic acid (6.0 mL) and reacted at 50 °C for 10 min. The mixture was concentrated under reduced pressure, and ethyl acetate (60 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate solution. The organic layer was separated and extracted with ethyl acetate (60 mL × 5). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 10:1) to give compound 25 (0.15 g, yield 64%).

¹ H NMR (400MHz, DMSO-d ₆ )δ8.84(d,1H),7.92(s,1H),7.80(dd,1H),7.73(d,1H),7.64–7.55(m,2H),7.48(t,1H),5.07(q,1H),4.51(s,2H),4.16(dd,1H),3. 94-3.84(m,1H),3.80-3.70(m,1H),3.45–3.13(m,3H),3.10(s,3H),3.02–2.91(m,1H),2.86–2.67(m,2H),1.90-1.74(m,2H).LC-MS m/z＝437.2[M+1] ⁺

Example 26: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(6-fluoro-3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 26)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(6-fluoro-3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamid(compound26)

Step 1: 6-Fluorobenzo[d]oxazol-2(3H)-one (26B)

6-fluorobenzo[d]oxazol-2(3H)-one(26B)

26A (10 g, 78.67 mmol) was dissolved in anhydrous DMF (130 mL), and N,N'-carbonyldiimidazole (15.31 g, 94.40 mmol) in DMF (100 mL) was added dropwise under ice bath conditions. After the addition was complete, the mixture was reacted at 60 °C for 4 h. After cooling to room temperature, 600 mL of water was added, and the mixture was extracted with EA (100 mL x 4). The combined organic layers were backwashed with saturated brine, dried over anhydrous sodium sulfate, and the residue was concentrated. The residue was purified by silica gel column chromatography (PE:EA (v/v) = 3:1) to give the title compound 26B (11 g, 91%). LC-MS (ESI): m/z = 154.1 [M+H] ⁺ .

Step 2: 6-Fluoro-5-iodobenzo[d]oxazol-2(3H)-one (26C)

6-fluoro-5-iodobenzo[d]oxazol-2(3H)-one(26C)

26B (3 g, 19.59 mmol) was dissolved in sulfuric acid (40 mL), and N-iodosuccinimide (5.29 g, 23.51 mmol) was added in portions. The reaction mixture was reacted overnight at 40 °C. After cooling to room temperature, the reaction mixture was poured into 500 mL of water and extracted with EA (50 mL x 5). The combined organic layers were backwashed with saturated sodium bicarbonate aqueous solution (200 mL), dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (PE:EA (v/v) = 4:1) to give the title compound 26C (1.1 g, 20%). LC-MS (ESI): m/z = 280.0 [M+H] ⁺ .

Step 3: 6-Fluoro-5-iodo-3-methylbenzo[d]oxazol-2(3H)-one (26D)

6-fluoro-5-iodo-3-methylbenzo[d]oxazol-2(3H)-one(26D)

26C (1 g, 3.58 mmol) was dissolved in acetonitrile (40 mL), potassium carbonate (0.59 g, 4.30 mmol) was added, and the mixture was stirred at 40 °C for 30 min. The mixture was then transferred to room temperature, and iodomethane (1.02 g, 7.16 mmol) was added dropwise. The reaction was allowed to proceed overnight at room temperature. Insoluble matter was removed by filtration, and the mixture was extracted with EA (50 mL x 3). The combined organic layers were backwashed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (PE:EA (v/v) = 5:1) to give the title compound 26D (0.92 g, 88%). LC-MS (ESI): m/z = 294.0 [M+H] ⁺ .

Step 4: 6-Fluoro-3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)benzo[d]oxazol-2(3H)-one (26E)

6-fluoro-3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]oxazol-2(3H)-one(26E)

26D (0.92 g, 3.14 mmol), pinacol diborate (0.96 g, 3.77 mmol), 1,1'-(diphenylphosphine)ferrocene]palladium dichlorochloride (0.23 g, 0.31 mmol), and potassium acetate (0.92 g, 9.42 mmol) were dissolved in dioxane (40 mL) and reacted at 100 °C for 5 h under nitrogen protection. After cooling to room temperature, 120 mL of water was added, and the mixture was extracted with EA (30 mL x 3). The combined organic layers were backwashed with saturated brine, dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (PE:EA (v/v) = 5:1) to give the title compound 26E (0.52 g, 56%).

¹ H NMR (400MHz, CDCl ₃ ) δ7.25 (d, J = 4.5 Hz, 1H), 6.95 (d, J = 8.2 Hz, 1H), 3.41 (s, 3H), 1.37 (s, 12H). LC-MS (ESI): m/z = 294.1 [M+H] ⁺ .

Step 5: (S)-(1-cyano-2-(2-fluoro-4-(6-fluoro-3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)tert-butyl carbamate (26F)

tert-butyl(S)-(1-cyano-2-(2-fluoro-4-(6-fluoro-3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)carbamate(26F)

Compound 2a (0.62 g, 1.81 mmol), 26E (0.53 g, 1.81 mmol), 1,1'-(diphenylphosphine)ferrocene]dichloropalladium (0.13 g, 0.18 mmol), and potassium carbonate (0.75 g, 5.43 mmol) were dissolved in a mixed solvent of dioxane (27 mL) and water (3 mL). The reaction was carried out at 90 °C for 5 h under nitrogen protection. After cooling to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic layers were combined and washed successively with saturated sodium bicarbonate (20 mL) and saturated brine (20 mL). The mixture was dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (PE:EA (v/v) = 2:1) to give the title compound 26F (0.6 g, 77%). LC-MS (ESI): m/z = 430.1 [M+H] ⁺ .

Step 6: (S)-2-amino-3-(2-fluoro-4-(6-fluoro-3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)propionitrile (26G)

(S)-2-amino-3-(2-fluoro-4-(6-fluoro-3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)propanenitrile(26G)

26F (0.6 g, 1.40 mmol) was dissolved in anhydrous formic acid (3 mL) and reacted at 50 °C for 20 min. After cooling to room temperature, most of the solvent was removed by concentration. A saturated sodium bicarbonate solution (20 mL) was added to the residue, and the mixture was extracted with EA (10 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 97:3) to give the title compound 26G (0.36 g, 78%). LC-MS (ESI): m/z = 330.0 [M+H] ⁺ .

Step 7: (S)-2-(((S)-1-cyano-2-(2-fluoro-4-(6-fluoro-3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (compound 26H)

tert-butyl(S)-2-(((S)-1-cyano-2-(2-fluoro-4-(6-fluoro-3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(compound 26H)

INT-3 (0.26 g, 1.06 mmol) was dissolved in DMF (5 mL). Under nitrogen protection, HATU (0.6 g, 1.59 mmol) and DIPEA (0.41 g, 3.18 mmol) were added. After stirring at room temperature for 20 min, 26 G (0.35 g, 1.06 mmol) was added, and the reaction was allowed to proceed for 1 h at room temperature. 30 mL of water was added to the reaction mixture, and the mixture was extracted with EA (15 mL x 5). The organic layers were combined, dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 97:3) to give the title compound 26H (0.33 g, 56%). LC-MS (ESI): m/z = 501.1 [M+H] ⁺ .

Step 8: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(6-fluoro-3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 26)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(6-fluoro-3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamid(compound26)

26H (0.33 g, 0.59 mmol) was dissolved in anhydrous formic acid (3 mL) and reacted at 50 °C for 1 h. After cooling to room temperature, most of the solvent was removed by concentration. A saturated sodium bicarbonate solution (25 mL) was added to the residue, and the mixture was extracted multiple times with EA until no product was found in the aqueous layer. The organic layers were combined, dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 10:1) to give compound 26 (120 mg, 44%).

¹ H NMR (400MHz, CD ₃ OD)δ7.47–7.22(m,5H),5.17(dd,J＝8.8,6.8Hz,1H),4.13(dd,J＝8.8,3.6 Hz,1H),4.02–3.95(m,1H),3.79(m,1H),3.43(s,3H),3.38–3.34(m,1H),3 .27–3.18(m,2H),2.99–2.91(m,1H),2.87–2.78(m,1H),2.67(dd,J＝14.4, 8.6Hz,1H),2.22–2.15(m,1H),2.03(m,1H).LC-MS(ESI):m/z＝457.1[M+H] ⁺ .

Example 27: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-(oxacyclobut-3-yl)-1,2,3,6-tetrahydropyridin-4-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-(oxetan-3-yl)-1,2,3,6-tetrahydropyridin-4-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

Step 1: 1-(oxetanebut-3-yl)-1,2,3,6-tetrahydropyridine-4-yl trifluoromethanesulfonate (27B)

1-(oxetan-3-yl)-1,2,3,6-tetrahydropyridin-4-yltrifluoromethanesulfonate

Under nitrogen protection at -78℃, LDA (2.6 mL, 5.2 mmol) was slowly added dropwise to 27A (400 mg, 2.6 mmol, preparation method referred to WO2016/172496A1) in THF (4.0 mL). After stirring for 30 minutes at this temperature, N,N-bis(trifluoromethanesulfonyl)aniline (1.40 g, 3.9 mmol) in THF solution (2.0 mL) was added dropwise. The mixture was allowed to rise naturally to room temperature for 1 hour. The reaction was quenched with 5 mL of water, extracted with ethyl acetate (5 mL × 3), and the organic phases were combined and washed with water (5 mL × 1), washed with saturated sodium chloride (5 mL × 1), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then subjected to column chromatography (DCM:MeOH = 50:1-20:1) to obtain a pale yellow liquid 27B (240 mg, 32.4%). LC-MS (ESI): m/z = 288.1 [M+H] ⁺ .

Step 2: (S)-(1-cyano-2-(2-fluoro-4-(1-(oxetane-3-yl)-1,2,3,6-tetrahydropyridin-4-yl)phenyl)ethyl)carboxylic acid tert-butyl ester (27C)

tert-butyl-(S)-(1-cyano-2-(2-fluoro-4-(1-(oxetan-3-yl)-1,2,3,6-tetrahydropyridin-4-yl)phenyl)ethyl)carbamate

INT-2 (200 mg, 0.6 mmol), 27B (200 mg, 0.7 mmol), Pd(dppf) _Cl₂ (44 mg, 0.06 mmol), and sodium carbonate (190 mg, 1.8 mmol) were added to a mixed solvent of 1,4-dioxane (5 mL) and water (1 mL) at room temperature. The mixture was purged with nitrogen three times and heated to 60°C for 2 hours. After cooling to room temperature, the reaction mixture was evaporated to dryness and purified by direct column chromatography (DCM:MeOH = 50:1-20:1) to give the title compound 27C as a yellow solid (180 mg, 64%). LC-MS (ESI): m/z = 402.2 [M+H] ^⁺ .

Step 3: (S)-2-amino-3-(2-fluoro-4-(1-(oxetanebut-3-yl)-1,2,3,6-tetrahydropyridin-4-yl)phenyl)propionitrile (27D)

(S)-2-amino-3-(2-fluoro-4-(1-(oxetan-3-yl)-1,2,3,6-tetrahydropyridin-4-yl)phenyl)propanenitrile

27C (180 mg, 0.4 mmol) was added to anhydrous formic acid (3.0 mL) at room temperature, and the mixture was heated to 50 °C and reacted for 20 minutes. Most of the formic acid was removed under reduced pressure, and the mixture was neutralized to alkaline by adding saturated sodium bicarbonate. The mixture was extracted with dichloromethane (10 mL × 4), washed with saturated sodium chloride (5 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude compound 27D, a yellow viscous substance (200 mg). LC-MS (ESI): m/z = 302.2 [M + H] ⁺ .

Step 4: (S)-2-(((S)-1-cyano-2-(2-fluoro-4-(1-(oxacyclobut-3-yl)-1,2,3,6-tetrahydropyridin-4-yl)phenyl)ethyl)formyl)-1,4-oxazacycloheptane-4-carboxylic acid tert-butyl ester (27E)

tert-butyl-(S)-2-(((S)-1-cyano-2-(2-fluoro-4-(1-(oxetan-3-yl)-1,2,3,6-tetrahydropyridin-4-yl)phenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate

Crude 27D (200 mg), INT-3 (170 mg, 0.7 mmol), and DIPEA (180 mg, 1.4 mmol) were added to dichloromethane at room temperature, followed by the addition of HATU (270 mg, 0.7 mmol). The mixture was stirred at room temperature for 1 hour. After the reaction mixture was evaporated to dryness, it was purified directly by column chromatography (DCM:MeOH = 50:1-20:1) to give a yellow solid 27E (200 mg, 94% yield in two steps). LC-MS (ESI): m/z = 529.3 [M+H] ⁺ .

Step 5: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-(oxacyclobut-3-yl)-1,2,3,6-tetrahydropyridin-4-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Chemical 27)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(1-(oxetan-3-yl)-1,2,3,6-tetrahydropyridin-4-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

27E (150 mg, 0.3 mmol) was added to anhydrous formic acid (3.0 mL) at room temperature, and the mixture was heated to 50 °C and reacted for 30 minutes. Most of the formic acid was removed under reduced pressure, and the mixture was neutralized to alkaline by adding saturated sodium bicarbonate. The mixture was extracted with dichloromethane (10 mL × 4), washed with saturated sodium chloride (5 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 27 (40 mg, 33%). LC-MS (ESI): m/z = 429.3 [M + H] ⁺ .

^1H NMR (400MHz, CDCl3 ₎ )δ7.27-7.21(m,2H),7.18-7.15(m,1H),7.12-7.08(m,1H),6.12-6.10(m,1H ),5.16-5.10(m,1H),4.74-4.68(m,4H),4.18-4.15(m,1H),4.05-3.99(m,1H) ,3.81-3.75(m,1H),3.68-3.63(m,1H),3.42-3.37(m,1H),3.22-3.14(m,2H) ,3.09-3.07(m,2H),3.01-2.92(m,3H),2.61-2.56(m,4H),1.95-1.92(m,3H).

Example 28: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(7-methoxy-1-methyl-1H-indol-4-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 28)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(7-methoxy-1-methyl-1H-indol-4-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

Step 1: 4-Bromo-7-methoxy-1-methyl-1H-indole (28B)

4-bromo-7-methoxy-1-methyl-1H-indole

28A (0.50 g, 2.21 mmol) was dissolved in acetonitrile (5 mL), and cesium carbonate (1.44 g, 4.22 mmol) and iodomethane (0.47 g, 3.31 mmol) were added. The mixture was reacted overnight at room temperature. Water (25 mL) was added, and the mixture was extracted with ethyl acetate (30 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 4:1) to give the title compound 28B as a white solid (0.50 g, 94.23%). LC-MS (ESI): m/z = 240.1 [M+H] ⁺ .

Step 2: (S)-(2-(4-(benzo[d]thiazolyl)-2-fluorophenyl)-1-cyanoethyl)tert-butyl carbamate (28C)

tert-butyl(S)-(2-(4-(benzo[d]thiazol-2-yl)-2-fluorophenyl)-1-cyanoethyl)carbamate

28B (0.40 g, 1.02 mmol) was dissolved in 1,4-dioxane (10 mL) and water (0.4 mL). INT-2 (0.26 g, 1.22 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloromethane complex (0.17 g, 0.20 mmol), and potassium carbonate (0.28 g, 2.04 mmol) were added. After the addition was complete, the mixture was heated to 90 °C for 2 h under nitrogen protection. The reaction was allowed to proceed overnight at room temperature. The mixture was concentrated to dryness, and saturated ammonium chloride aqueous solution (50 mL) was added. The mixture was extracted with ethyl acetate (20 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 4:1) to give the title compound 28C, a pale yellow liquid (0.24 g, 59.20%). LC-MS (ESI): m/z=398.1[M+H] ⁺ .

Step 3: (S)-2-amino-3-(4-(benzo[d]thiazolyl)-2-fluorophenyl)propionitrile (28D)

(S)-2-amino-3-(4-(benzo[d]thiazol-2-yl)-2-fluorophenyl)propanenitrile

28C (0.32 g, 0.81 mmol) was dissolved in formic acid (5 mL), and the reaction was carried out at 30 °C for 3 h after the addition was complete. The pH was adjusted to approximately 8 by dropwise addition of saturated sodium bicarbonate aqueous solution, and the mixture was extracted with ethyl acetate (25 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 28D as a colorless liquid (0.24 g, 99.65%), which was used directly in the next reaction. LC-MS (ESI): m/z = 298.1 [M+H] ⁺ .

Step 4: (S)-2-(((S)-2-(4-(benzo[d]thiazolyl)-2-fluorophenyl)-1-cyanoethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (28E)

tert-butyl(S)-2-(((S)-2-(4-(benzo[d]thiazol-2-yl)-2-fluorophenyl)-1-cyanoethyl)carbamoyl)-1,4-oxazepane-4-carboxylate

28D (0.24 g, 0.81 mmol) was dissolved in dichloromethane (10 mL), and INT-3 (0.26 g, 1.05 mmol), diisopropylethylamine (0.31 g, 2.43 mmol), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (0.46 g, 1.22 mmol) were added. The mixture was reacted overnight at room temperature. A saturated sodium chloride aqueous solution (30 mL) was added, and the mixture was extracted with ethyl acetate (25 mL). The organic phase was washed with saturated sodium chloride aqueous solution (25 mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 4:1) to give title compound 28E, a pale yellow solid (0.25 g, 58.83%). LC-MS (ESI): m/z = 469.2 [M-57+H] ⁺ .

Step 5: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(7-methoxy-1-methyl-1H-indol-4-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 28)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(7-methoxy-1-methyl-1H-indol-4-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

28E (0.32 g, 0.59 mmol) was dissolved in formic acid (2.0 mL), and the mixture was reacted at 35 °C for 4 h after the addition was complete. The solution was concentrated to dryness, and ethyl acetate (25 mL) was added. The pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with ethyl acetate (25 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 20:1) to give the title compound 28 (30 mg, 14.72%).

¹ H NMR (400MHz, CDCl ₃ )δ7.49–7.28(m,3H),7.04–6.88(m,2H),6.67(d,1H),6.54(d,1H),5.32–5.04(m,1H),4.12–4.05(m,4H),4.01 –3.89(m,4H),3.75(m,1H),3.26(qd,3H),2.97–2.82(m,3H),1.90–1.68(m,2H).LC-MS(ESI):m/z＝451.2[M+H] ⁺ .

Example 29: (S)-N-((S)-1-cyano-2-(4'-cyano-3-fluoro-3'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 29)

(S)-N-((S)-1-cyano-2-(4'-cyano-3-fluoro-3'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 29)

Step 1: (S)-(1-cyano-2-(4'-cyano-3-fluoro-3'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)ethyl)tert-butyl carbamate (29B)

tert-butyl(S)-(1-cyano-2-(4'-cyano-3-fluoro-3'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)ethyl)carba mate(29B)

Compound 29A (300.0 mg, 1.00 mmol), INT-2 (470.0 mg, 1.20 mmol), Pd(dppf) _Cl₂ (160.0 mg, 0.20 mmol), and potassium carbonate (280.0 mg, 2.00 mmol) were added to a single-necked flask, followed by 1,4-dioxane (10 mL) and water (0.4 mL). After purging with nitrogen three times, the mixture was reacted at 95 °C for 4 hours. After cooling to room temperature, the mixture was concentrated and purified by silica gel column chromatography (PE:EA(v/v) = 2:1) to give the title compound 29B as a white solid (230.0 mg, 53.1%). LC-MS (ESI): m/z = 377.1 [M-57+H] ^⁺ .

Step 2: (S)-4'-(2-amino-2-cyanoethyl)-3'-fluoro-3-(trifluoromethyl)-[1,1'-biphenyl]-4-nitrile (29C)

(S)-4'-(2-amino-2-cyanoethyl)-3'-fluoro-3-(trifluoromethyl)-[1,1'-biphenyl]-4-carbonitrile(29C)

29B (230.0 mg, 0.53 mmol) was dissolved in formic acid (5 mL) and reacted at 35 °C for 4 h. The reaction mixture was adjusted to alkaline with saturated potassium carbonate solution, extracted with ethyl acetate (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 29C, a pale yellow oil (180.0 mg, 100%), which was used directly in the next reaction. LC-MS (ESI): m/z = 334.1 [M+H] ⁺ .

Step 3: (S)-2-(((S)-1-cyano-2-(4'-cyano-3-fluoro-3'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (29D)

tert-butyl(S)-2-(((S)-1-cyano-2-(4'-cyano-3-fluoro-3'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(29D)

29C (180 mg, 0.54 mmol) was dissolved in DMF (5 mL), and INT-3 (160 mg, 0.65 mmol), HATU (250 mg, 0.65 mmol), and DIPEA (210 mg, 1.62 mmol) were added. The mixture was reacted overnight at room temperature. Water (20 mL) was added to the system, and the mixture was extracted with ethyl acetate (30 mL × 2). The extract was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (PE:EA (v/v) = 2:1) to give the title compound 29D as a white solid (280 mg, 92.5%). LC-MS (ESI): m/z = 505.2 [M-57+H] ⁺ .

Step 4: (S)-N-((S)-1-cyano-2-(4'-cyano-3-fluoro-3'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 29)

(S)-N-((S)-1-cyano-2-(4'-cyano-3-fluoro-3'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 29)

29D (280 mg, 0.50 mmol) was dissolved in formic acid (5 mL) and reacted at 35 °C for 4 h. The reaction mixture was adjusted to alkaline with saturated potassium carbonate solution, extracted with ethyl acetate (30 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 30:1) to give title compound 29 (100.0 mg, 43.4%).

¹ H NMR (400MHz, CDCl ₃ )δ7.96–7.92(m,2H),7.85(dd,1H),7.49(t,1H),7.40(dd,1H),7.35(dd,1H),5.23–5.17(m,1H),4.09(q,1H),4.05–4.00(m,1H),3.80 –3.74(m,1H),3.30(dd,1H),3.29–3.20(m,2H),2.98–2.86(m,3H),1.93–1.80(m,2H),1.61–1.50(m,2H).LC-MS(ESI):m/z＝461.2[M+H] ⁺ .

Example 30: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(3-oxoisoindoline-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 30)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(3-oxoisoindolin-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

Step 1: (S)-(1-cyano-2-(2-fluoro-4-(3-oxoisoindoline-5-yl)phenyl)ethyl)tert-butyl carbamate (30B)

tert-butyl(S)-(1-cyano-2-(2-fluoro-4-(3-oxoisoindolin-5-yl)phenyl)ethyl)carbamate

30A (0.20 g, 0.95 mmol), INT-2 (0.31 g, 0.79 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (0.12 g, 0.16 mmol), and potassium carbonate (0.22 g, 1.58 mmol) were added sequentially to 1,4-dioxane (20 mL) and water (4 mL). The system was purged three times with nitrogen and reacted at 100 °C for 2.5 h. After the reaction was complete, the mixture was cooled to room temperature, and water (40 mL) was added. The aqueous phase was extracted with ethyl acetate (60 mL × 3). The combined organic phases were washed with saturated sodium chloride aqueous solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by silica gel column chromatography (PE:EA (v/v) = 3:1 to 2:1) to give 30B (0.26 g, yield 83%). LCMS m/z = 396.1[M+H] ⁺ .

Step 2: (S)-2-amino-3-(2-fluoro-4-(3-oxoisoindoline-5-yl)phenyl)propionitrile (30C)

(S)-2-amino-3-(2-fluoro-4-(3-oxoisoindolin-5-yl)phenyl)propanenitrile

30B (0.26 g, 0.66 mmol) was dissolved in formic acid (5.0 mL), and the mixture was reacted at 50 °C for 10 minutes after the addition was complete. The solution was concentrated to dryness, and ethyl acetate (60 mL) was added. The pH was adjusted to approximately 8 by dropwise addition of saturated sodium bicarbonate solution. The organic layer was separated, and the aqueous phase was extracted with ethyl acetate (60 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 30C (0.15 g, 77% yield). LCMS m/z = 296.1 [M+H] ⁺ .

Step 3: (S)-2-(((S)-1-cyano-2-(2-fluoro-4-(3-oxoisoindoline-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (30D)

tert-butyl(S)-2-(((S)-1-cyano-2-(2-fluoro-4-(3-oxoisoindolin-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate

30C (0.15 g, 0.51 mmol) was dissolved in N,N-dimethylformamide (10 mL), and INT-3 (0.13 g, 0.51 mmol), triethylamine (0.1 g, 1 mmol), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (0.23 g, 0.61 mmol) were added. The mixture was reacted at room temperature for 1 hour after the addition was complete. Saturated sodium chloride aqueous solution (30 mL) was added, and the mixture was extracted with ethyl acetate (60 mL × 3). The organic phase was washed with saturated sodium chloride aqueous solution (60 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated by silica gel column chromatography (PE:EA (v/v) = 2:1 to 1:5) to give the title compound 30D (0.14 g, yield 53%).

Step 4: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(3-oxoisoindoline-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 30)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(3-oxoisoindolin-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

30D (0.14 g, 0.27 mmol) was dissolved in formic acid (6.0 mL) and reacted at 50 °C for 10 min. The mixture was concentrated under reduced pressure, ethyl acetate (60 mL) was added, and the pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with dichloromethane (60 mL × 5). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 4:1) to give compound 30 (48 mg, yield 42%).

¹ H NMR (400MHz, DMSO-d ₆ )δ8.93(d,1H),8.62(s,1H),7.98-7.92(m,2H),7.71–7.55(m,3H),7.47(t,1H),5.06(q,1H),4.42(s,2H),4.27(dd,1H),3.95- 3.90m,1H),3.80-3.72(m,1H),3.28–3.15(m,3H),3.12-3.02(m,1H),2.95–2.87(m,1H),2.81(dd,1H),1.94-1.84(m,2H).LC-MS m/z=423.2[M+H] ⁺ .

Example 31: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(2-methyl-3-oxoisoindoline-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 31)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(2-methyl-3-oxoisoindolin-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

Step 1: 6-Bromo-2-methyl-isoindoline-1-one (31A)

6-bromo-2-methylisoindolin-1-one

30A (0.49 g, 2.3 mmol) was dissolved in dry N,N-dimethylformamide (20 mL), cooled to 0 °C under nitrogen protection, and sodium hydride (0.14 g, 3.45 mmol, 60% wt) was added in portions. After the addition was complete, the reaction was allowed to proceed for 20 minutes, and then iodomethane (0.49 g, 3.45 mmol) was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature for 30 minutes. The reaction was quenched with water (100 mL), extracted with ethyl acetate (100 mL × 2), the organic phases were combined, washed with saturated brine (100 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by silica gel column chromatography (PE:EA = 6:1) to give the target compound 31A (0.37 g, 71% yield).

¹ H NMR (400MHz, CDCl ₃ ) δ7.97(d,1H),7.64(dd,1H),7.31(d,1H),4.33(s,2H),3.20(s,3H).

Step 2: (S)-(1-cyano-2-(2-fluoro-4-(2-methyl-3-oxoisoindoline-5-yl)phenyl)ethyl)tert-butyl carbamate (31B)

tert-butyl(S)-(1-cyano-2-(2-fluoro-4-(2-methyl-3-oxoisoindolin-5-yl)phenyl)ethyl)carbamate

31A (0.18 g, 0.8 mmol), INT-2 (0.29 g, 0.74 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (0.11 g, 0.15 mmol), and potassium carbonate (0.2 g, 1.48 mmol) were added sequentially to 1,4-dioxane (20 mL) and water (4 mL). The system was purged three times with nitrogen and reacted at 100 °C for 2 hours. After the reaction was complete, the mixture was cooled to room temperature, and water (40 mL) was added. The aqueous phase was extracted with ethyl acetate (60 mL × 3). The combined organic phases were washed with saturated sodium chloride aqueous solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by silica gel column chromatography (PE:EA (v/v) = 3:1 to 1:1) to give 31B (0.27 g, yield 89%). LCMS m/z = 432.1 [M+Na] ⁺ .

Step 3: (S)-2-amino-3-(2-fluoro-4-(2-methyl-3-oxoisoindoline-5-yl)phenyl)propionitrile (31C)

(S)-2-amino-3-(2-fluoro-4-(2-methyl-3-oxoisoindolin-5-yl)phenyl)propanenitrile

31B (0.27 g, 0.66 mmol) was dissolved in formic acid (6.0 mL), and the mixture was reacted at 50 °C for 10 minutes after the addition was complete. The solution was concentrated to dryness, and ethyl acetate (60 mL) was added. The pH was adjusted to approximately 8 by dropwise addition of saturated sodium bicarbonate solution. The organic layer was separated, and the aqueous phase was extracted with ethyl acetate (60 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 31C (0.16 g, 78% yield). LCMS m/z = 310.2 [M+H] ⁺ .

Step 4: (S)-2-(((S)-1-cyano-2-(2-fluoro-4-(2-methyl-3-oxoisoindoline-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazacycloheptane-4-carboxylic acid tert-butyl ester (31D)

tert-butyl tert-butyl(S)-2-(((S)-1-cyano-2-(2-fluoro-4-(2-methyl-3-oxoisoindolin-5-yl)phenyl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate

31C (0.16 g, 0.52 mmol) was dissolved in N,N-dimethylformamide (10 mL), and INT-3 (0.13 g, 0.52 mmol), triethylamine (0.1 g, 1 mmol), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (0.24 g, 0.62 mmol) were added. The mixture was reacted at room temperature for 1 hour. Saturated sodium chloride aqueous solution (30 mL) was added, and the mixture was extracted with ethyl acetate (60 mL × 3). The organic phase was washed with saturated sodium chloride aqueous solution (60 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated by silica gel column chromatography (PE:EA (v/v) = 2:1 to 1:3) to give the title compound 31D (0.21 g, 75% yield). LCMS m/z = 559.2 [M+Na] ⁺ .

Step 5: (S)-N-((S)-1-cyano-2-(2-fluoro-4-(2-methyl-3-oxoisoindoline-5-yl)phenyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 31)

(S)-N-((S)-1-cyano-2-(2-fluoro-4-(2-methyl-3-oxoisoindolin-5-yl)phenyl)ethyl)-1,4-oxazepane-2-carboxamide

31D (0.21 g, 0.39 mmol) was dissolved in formic acid (6.0 mL) and reacted at 50 °C for 10 min. The mixture was concentrated under reduced pressure, ethyl acetate (60 mL) was added, and the pH was adjusted to approximately 8 by adding saturated sodium bicarbonate aqueous solution. The organic layer was separated and extracted with dichloromethane (60 mL × 5). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 10:1) to give compound 31 (100 mg, yield 59%).

¹ H NMR (400MHz, DMSO-d ₆ )δ8.85(d,1H),7.94-7.90(m,2H),7.68(d,1H),7.65–7.56(m,2H),7.46(t,1H),5.11–5.01(m,1H),4.51(s,2H),4.18(dd,1H),3. 94-3.84(m,1H),3.79-3.70(m,1H),3.26–3.14(m,3H),3.10(s,3H),3.02-2.92(m,1H),2.87–2.69(m,2H),1.90-1.77m,2H).LC-MS m/z=437.2[M+H] ⁺ .

Example 32: (2S)-N-(1-cyano-2-(3-fluoro-5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophene-2-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 32)

(2S)-N-(1-cyano-2-(3-fluoro-5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophen-2-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 32)

Step 1: Methyl 5-bromo-3-fluorothiophene-2-carboxylic acid (32B)

methyl 5-bromo-3-fluorothiophene-2-carboxylate(32B)

At room temperature, 32A (15 g, 93.7 mmol) was dissolved in chloroform (200 mL), bromine (120 g, 750 mmol) was added, and the mixture was heated to 80 °C for 3 hours. The reaction solution was then poured into a saturated sodium thiosulfate solution (500 mL), the organic phase was concentrated to dryness, and the mixture was purified to obtain the title compound 32B (10 g, 45% yield).

Step 2: (5-bromo-3-fluorothiophene-2-yl)methanol (32C)

(5-bromo-3-fluorothiophen-2-yl)methanol(32C)

At room temperature, 32B (5 g, 20.9 mmol) was dissolved in dichloromethane (120 mL), and diisobutylaluminum hydride (42 mL, 62.8 mmol) was added dropwise. The mixture was reacted at room temperature for 3 hours, and then water (100 mL) was added. The mixture was filtered, and the organic phase was concentrated to dryness to give the title compound 32C (4 g, 90% yield).

Step 3: 5-Bromo-2-(Bromomethyl)-3-fluorothiophene (32D)

5-bromo-2-(bromomethyl)-3-fluorothiophene(32D)

At room temperature, 32C (4 g, 19.0 mmol) was dissolved in dichloromethane (60 mL), and carbon tetrabromide (7.5 g, 22.7 mmol) and triphenylphosphine (7.5 g, 28.4 mmol) were added. The mixture was reacted at room temperature for 2 hours. The reaction solution was concentrated to dryness and purified by direct column chromatography (PE:EA(v:v) = 20:1-10:1) to give the title compound 32D (4 g, 77% yield).

Step 4: 3-(5-bromo-3-fluorothiophene-2-yl)-2-((diphenylmethylene)amino)propionitrile (32E)

3-(5-bromo-3-fluorothiophen-2-yl)-2-((diphenylmethylene)amino)propanenitrile(32E)

32D (2 g, 7.3 mmol) was dissolved in dichloromethane (40 mL) and water (4 mL) at room temperature. Sodium hydroxide (0.5 g, 13.1 mmol) and benzyltrimethylammonium chloride (140 mg, 0.73 mmol) were added, and the mixture was reacted at room temperature for 16 hours. The reaction solution was washed with water (50 mL × 1), the organic phase was concentrated, and the mixture was purified by direct column chromatography (PE:EA(v:v) = 20:1-10:1) to give the title compound 32E (1.2 g, 40% yield). LC-MS (ESI): m/z = 413.1 [M+H] ⁺ .

Step 5: 2-((diphenylmethylene)amino)-3-(3-fluoro-5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophen-2-yl)propionitrile (32F)

2-((diphenylmethylene)amino)-3-(3-fluoro-5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophen-2-yl)propanenitrile(32F)

At room temperature, 32E (1.2 g, 2.9 mmol) was dissolved in dioxane (30 mL) and water (3 mL), followed by the addition of 1A (1 g, 3.5 mmol), potassium carbonate (1 g, 7.3 mmol), 2-dicyclohexylphospho-2,4,6-triisopropylbiphenyl (0.28 g, 0.6 mmol), and palladium acetate (0.065 g, 0.29 mmol). The reaction mixture was reacted at 100 °C for three hours. The reaction solution was then directly mixed and purified by column chromatography (PE:EA(v:v) = 5:1-1:1) to give the title compound 32F (1.4 g, 70% yield). LC-MS (ESI): m/z = 482.1 [M+H] ⁺ .

Step 6: 2-Amino-3-(3-fluoro-5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophen-2-yl)propionitrile (32G)

2-amino-3-(3-fluoro-5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophen-2-yl)propane-nitrile(32G)

At room temperature, 32F (1.4 g, 2.9 mmol) was dissolved in tetrahydrofuran (20 mL), and 1 N hydrochloric acid (40 mL) was added. The reaction was carried out at room temperature for 1 hour. The reaction solution was extracted with ethyl acetate (50 mL × 1), and the pH of the aqueous phase was adjusted to 8-9 with potassium carbonate. The product was extracted with ethyl acetate (50 mL × 2), and the organic phase was concentrated to give the title compound 32G (0.45 g, 49% yield). LC-MS (ESI): m/z = 318.1 [M+H] ⁺ .

Step 7: (2S)-2-((1-cyano-2-(3-fluoro-5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophene-2-yl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (32H)

(2S)-tert-butyl-2-((1-cyano-2-(3-fluoro-5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophen-2-yl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(32H)

At room temperature, 32 g (0.45 g, 1.4 mmol) was dissolved in N,N-dimethylformamide (10 mL), followed by N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (0.6 g, 1.6 mmol), INT-3 (0.4 g, 1.7 mmol), and diisopropylethylamine (0.5 g, 3.6 mmol). The reaction mixture was reacted at room temperature for 2 hours. The reaction solution was poured into water (200 mL), extracted with ethyl acetate (100 mL × 2), and the organic phase was concentrated to dryness to give the title compound 32H (0.4 g, yield 52%). LC-MS (ESI): m/z = 545.2 [M + H] ⁺

Step 8: (2S)-N-(1-cyano-2-(3-fluoro-5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophene-2-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compound 32)

(2S)-N-(1-cyano-2-(3-fluoro-5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)thiophen-2-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 32)

At room temperature, compound 32G (0.4 g, 0.73 mmol) was dissolved in acetonitrile (20 mL), p-toluenesulfonic acid (0.5 g, 3.0 mmol) was added, and the reaction was carried out at 50 °C for 1 hour. The reaction solution was poured into saturated sodium bicarbonate aqueous solution (100 mL), extracted with ethyl acetate (100 mL × 2), the organic phase was concentrated, and purified by column chromatography (DCM:MeOH (v:v) = 50:1-20:1) to give compound 32 (0.24 g, 70%).

¹ H NMR (400MHz, CDCl ₃ )δ7.33-7.29(s,1H),7.21-7.19(m,1H),7.07-7.03(m,1H),7.00(s,1H),5.18-5.13(m,1H),4.14-4.00(m,2H),3.8 1-3.71(m,1H),3.41(s,3H),3.36-3.23(m,3H),3.04-2.90(m,3H),1.91-1.81(m,2H).LC-MS(ESI):m/z＝445.1[M+H] ⁺ .

Examples 33 and 34: (S)-N-((S)-1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-2-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide and (S)-N-((R)-1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-2-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compounds 33 and 34)

(S)-N-((S)-1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-2-yl)ethyl)-1,4-oxazepane-2-carboxamide and(S)-N-((R)-1 -cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-2-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 33 and compound 34)

Step 1: 5-Bromo-2-(Bromomethyl)pyridine (33B)

5-bromo-2-(bromomethyl)pyridine(33B)

2-Methyl-5-bromopyridine 33A (5 g, 29.07 mmol) was dissolved in carbon tetrachloride (50 mL), and N-bromosuccinimide (5.43 g, 30.52 mmol) and azobisisobutyronitrile (1.19 g, 7.27 mmol) were added. The mixture was reacted at 90 °C for 2.5 h. After cooling to room temperature, the reaction solution was concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 10:1) to give the title compound 33B (4.42 g, 61%). LC-MS (ESI): m/z = 251.9 [M+H] ⁺ .

Step 2: 3-(5-bromopyridin-2-yl)-2-((diphenylmethylene)amino)propionitrile (33C)

3-(5-bromopyridin-2-yl)-2-((diphenylmethylene)amino)propanenitrile(33C)

33B (4.42 g, 17.54 mmol) and N-(diphenylmethylene)aminoacetonitrile (3.86 g, 17.54 mmol) were dissolved in dichloromethane (50 mL). Benzyltrimethylammonium chloride (0.33 g, 1.75 mmol) was added, followed by the addition of an aqueous solution of sodium hydroxide (1.40 g, 35.08 mmol) (5 mL) with vigorous stirring. The reaction was allowed to proceed overnight at room temperature. 100 mL of water was added, and the mixture was extracted with dichloromethane (30 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (PE:EA (v/v) = 10:1) to give the title compound 33C (5.20 g, 76%). LC-MS (ESI): m/z = 390.0 [M+H] ⁺ .

Step 3: 2-((diphenylmethylene)amino)-3-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)propionitrile (33D)

2-((diphenylmethylene)amino)-3-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)propanenitrile(33D)

33C (1.73 g, 4.43 mmol), 1A (1.34 g, 4.87 mmol), 1,1'-(diphenylphosphine)ferrocene]dichloropalladium (0.33 g, 0.44 mmol), and potassium carbonate (1.22 g, 8.86 mmol) were dissolved in a mixed solvent of dioxane (50 mL) and water (5 mL). The reaction was carried out at 90 °C for 5 h under nitrogen protection. After cooling to room temperature, 200 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic layers were combined and washed successively with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL). The mixture was dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (PE:EA (v/v) = 3:1) to give the title compound 33D (2.03 g, 68.92%). LC-MS (ESI): m/z = 459.1 [M+H] ⁺ .

Step 4: 2-Amino-3-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-2-yl)propionitrile (33E)

2-amino-3-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-2-yl)propanenitrile(33E)

33D (1.02 g, 2.19 mmol) was dissolved in tetrahydrofuran (50 mL) and water (5 mL), and 2.5 mL of 1 M HCl aqueous solution was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature for 5 h. The reaction mixture was extracted with diethyl ether (15 mL x 3) and discarded. The aqueous layer was adjusted to pH 12 with 2 M NaOH aqueous solution and extracted with DCM (20 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated to obtain 580 mg of crude 33E, the title compound. This crude product was used directly in the next reaction without further purification. LC-MS (ESI): m/z = 295.0 [M+H] ⁺ .

Step 5: (2S)-2-((1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-2-yl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (33F)

tert-butyl(2S)-2-((1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-2-yl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate

33E (0.4 g, 1.36 mmol) was dissolved in dichloromethane (10 mL), and DIPEA (0.34 g, 2.64 mmol), HATU (0.55 g, 1.45 mmol), and intermediate INT-3 (0.36 g, 1.5 mmol) were added sequentially. The reaction was carried out at room temperature for 1 hour. After the reaction was confirmed to be complete by TLC, water (20 mL) was added to the reaction solution, and the layers were separated. The organic phase was washed successively with water (20 mL) and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a yellow oily crude product. The crude product was purified by rapid column chromatography (DCM:MeOH (v/v) = 96:4) to give a white solid 33F (0.4 g, yield 56.4%). LC-MS (ESI): m/z = 522.2 [M+H] ⁺ .

Step 6: (2S)-N-(-1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-2-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (33G)

(2S)-N-(1-cyano-2-(5-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-2-yl)ethyl)-1,4-oxazepane-2-carboxamide

Compound 33F (0.3 g, 0.58 mmol) was dissolved in dichloromethane (10 mL), and TMSOTf (0.19 g, 0.87 mmol) was added. 2,6-lutidine (0.12 g, 1.16 mmol) was added dropwise under ice bath conditions. After the addition was complete, the mixture was heated to room temperature and reacted for 1 h. The reaction solution was poured into 30 mL of saturated ammonium chloride solution and extracted with dichloromethane (20 mL × 3). The organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated to obtain a pale yellow oily crude product. The crude product was purified by rapid column chromatography (DCM:MeOH = 95:5, v/v) to obtain compound 33G (0.14 g).

Compound 33G was separated by chiral preparation via SFC to obtain peak 1 (40 mg, ee% = 100%, yield 16.5%, retention time: 2.728 min, designated as compound 33) and peak 2 (50 mg, ee% = 98.5%, yield: 20.6%, retention time: 3.987 min, designated as compound 34).

The purification conditions were as follows: (Instrument name: MG II preparative SFC (SFC-14; Column: ChiralPak AD, 250×30mm ID, 10μm; Mobile phase: Phase A: _CO2 ; Phase B: Isopropanol (0.1% _NH3 · _H2O ); Flow rate: 70mL/min; Column pressure: 100bar; Column temperature: 35℃; Absorption wavelength: 220nm; Cycle time: approximately 7min).

Peak 1: LC-MS (ESI): m/z=422.2[M+H] ⁺ . ;

¹ H NMR (400MHz, DMSO-d ₆ )δ8.79-8.78(m,1H),8.50-8.48(m,1H),7.88-7.85(m,1H),7.37-7.31(m,3H),7.14(s,1H),5.37-5.35(m,1H),4.28-4.25(m,1H),4.12-4. 08(m,1H),3.85-3.69(m,1H),3.52-3.51(m,1H),3.47(s,3H),3.37-3 .35(m,2H),3.13-3.05(m,2H),2.02-2.01(m,2H),1.26-1.20(m,2H).

Peak 2: LC-MS (ESI): m/z=422.2[M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ )δ8.79-8.78(m,1H),8.50-8.48(m,1H),7.88-7.85(m,1H),7.37-7.31(m,3H),7.14(s,1H),5.39-5.34(m,1H),4.28-4.25(m,1H),4.15-4. 09(m,1H),3.85-3.71(m,1H),3.52-3.51(m,1H),3.47(s,3H),3.37-3 .34(m,2H),3.14-3.04(m,2H),2.02-2.01(m,2H),1.26-1.20(m,2H).

Examples 35 and 36: (S)-N-((S)-1-cyano-2-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide and (S)-N-((R)-1-cyano-2-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compounds 35 and 36)

(S)-N-((S)-1-cyano-2-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazepane-2-carboxamide and(S)-N-((R)-1 -cyano-2-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 35 and compound 36)

Step 1: 2-Bromo-4-fluoro-5-methylpyridine (35B)

2-bromo-4-fluoro-5-methylpyridine

2-Bromo-4-amino-5-methylpyridine 35A (1.87 g, 10 mmol) was dissolved in pyridine hydrofluoric acid (20 mL). Sodium nitrite (0.83 g, 12 mmol) was added in portions at -10 °C. After the addition was complete, the mixture was allowed to rise to room temperature and reacted overnight. After the reaction was complete, water (100 mL) was added, the pH was adjusted to 8 with saturated sodium bicarbonate, and the mixture was extracted with ethyl acetate (100 mL x 2). The extract was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by silica gel column chromatography (PE:EA = 10:1, v/v) to give the target compound 35B (1.21 g, yield 63.4%).

¹ H NMR (400MHz, CDCl ₃ ) δ8.20(d,1H),7.19(d,1H),2.23(s,3H).LC-MS m/z＝190.0/192.0[M+1] ⁺

Step 2: 2-Bromo-5-(bromomethyl)-4-fluoropyridine (35°C)

2-bromo-5-(bromomethyl)-4-fluoropyridine

35B (2.84 g, 15.0 mmol) was dissolved in carbon tetrachloride (50 mL), and N-bromosuccinimide (2.93 g, 16.5 mmol) and azobisisobutyronitrile (0.49 g, 3.0 mmol) were added. The mixture was reacted at 90 °C for 4 h. After cooling to room temperature, the reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v/v) = 10:1) to give the title compound 35C (2.45 g, 61%).

¹ H NMR (400MHz, CDCl ₃ ) δ8.40(d,1H),7.28(d,1H),4.43(s,2H).

Step 3: 3-(6-bromo-4-fluoropyridin-3-yl)-2-((diphenylmethylene)amino)propionitrile (35D)

3-(6-bromo-4-fluoropyridin-3-yl)-2-((diphenylmethylene)amino)propanenitrile

35C (2.4 g, 8.9 mmol) and N-(diphenylmethylene)aminoacetonitrile (1.96 g, 8.9 mmol) were dissolved in dichloromethane (40 mL). Benzyltrimethylammonium chloride (0.17 g, 0.89 mmol) was added, followed by the addition of an aqueous solution of sodium hydroxide (1.07 g, 26.8 mmol) (4 mL) with vigorous stirring. The reaction was allowed to proceed overnight at room temperature. 100 mL of water was added, and the mixture was extracted with dichloromethane (60 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (PE:EA (v/v) = 10:1) to give the title compound 35D (2.23 g, 61%). LC-MS (ESI): m/z = 408.1 [M+H] ⁺ .

Step 4: 2-((diphenylmethylene)amino)-3-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridinyl)propionitrile (35E)

2-((diphenylmethylene)amino)-3-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)propanenitrile(35E)

35D (1.3 g, 3.18 mmol), 1A (0.87 g, 3.18 mmol), 1,1'-(diphenylphosphine)ferrocene]dichloropalladium (0.35 g, 0.48 mmol), and potassium carbonate (0.88 g, 6.36 mmol) were dissolved in a mixed solvent of dioxane (30 mL) and water (5 mL). The reaction was carried out at 90 °C for 5 h under nitrogen protection. After cooling to room temperature, 80 mL of water was added, and the mixture was extracted with ethyl acetate (60 mL x 3). The organic layers were combined and washed successively with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL). The mixture was dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (PE:EA (v/v) = 3:1) to give the title compound 35E (1.02 g, 67%). LC-MS (ESI): m/z = 477.2 [M+H] ⁺ .

Step 5: 2-Amino-3-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)propionitrile (35F)

2-amino-3-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)propanenitrile(35F)

35E (1.02 g, 2.14 mmol) was dissolved in tetrahydrofuran (25 mL) and water (5 mL), and 5 mL of 1M HCl aqueous solution was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature for 5 h. The reaction mixture was extracted with diethyl ether (40 mL) and discarded. The aqueous layer was adjusted to pH 12 with 2M NaOH aqueous solution and extracted with DCM (50 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated to obtain 0.64 g of crude 35F, the title compound. This crude product was used directly in the next reaction without further purification. LC-MS (ESI): m/z = 313.1 [M+H] ⁺ .

Step 6: (2S)-N-(1-cyano-2-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (35G)

(2S)-N-(1-cyano-2-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazepane-2-carboxamide

35F (0.64 g, 2.05 mmol) was dissolved in dichloromethane (10 mL), and intermediates INT-3 (0.5 g, 2.05 mmol), DIPEA (0.53 g, 4.1 mmol), and HATU (0.94 g, 2.46 mmol) were added sequentially. The reaction was carried out at room temperature for 1 hour. After the reaction was confirmed to be complete by TLC, water (20 mL) was added to the reaction solution, and the layers were separated. The organic phase was washed successively with water (20 mL) and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a yellow oily crude product. The crude product was purified by rapid column chromatography (DCM:MeOH (v/v) = 20:1) to give 35G (0.98 g, yield 88.6%). LC-MS (ESI): m/z = 540.2 [M+H] ⁺ .

Step 7: (S)-N-((S)-1-cyano-2-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide and (S)-N-((R)-1-cyano-2-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (compounds 35 and 36)

(S)-N-((S)-1-cyano-2-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazepane-2-carboxamide a nd(S)-N-((R)-1-cyano-2-(4-fluoro-6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazepane-2-carboxamide

Compound 35G (0.98 g, 1.82 mmol) was dissolved in formic acid (10 mL) and reacted at 50 °C for 10 min. The solution was concentrated under reduced pressure, and dichloromethane (40 mL) and saturated sodium bicarbonate (40 mL) were added. The mixture was separated, and the aqueous phase was extracted with dichloromethane (50 mL × 4). The combined organic phases were washed with saturated sodium chloride (40 mL), dried over anhydrous sodium sulfate, and concentrated to give the target compound (0.76 g, 95% yield). Two isomers were prepared by SFC: peak 1 (retention time: 2.47 min, designated as compound 35) and peak 2 (retention time: 3.69 min, designated as compound 36).

Preparation conditions: Instrument: MG II preparative SFC (SFC _- 14). Column: ChiralPak AD, 250×30mm ID, 10μm. Mobile phase: A: _CO2 , B: methanol (0.1% _NH3H2O ). Gradient: B 40%. Flow rate: 80mL/min. Back pressure: 100bar. Column temperature: 38℃. Wavelength: 220nm. Cycle time: approximately 10min. Sample preparation: Sample dissolved in 15ml methanol/dichloromethane. Injection: 3.5ml per syringe.

Peak 1: LC-MS m/z=440.1[M+1] ⁺ .

¹ H NMR (400MHz, CDCl ₃ )δ8.60(d,1H),7.72(d,1H),7.67(dd,1H),7.47(d,1H),7.32–7.26(m,2H),5.28–5.17(m,1H),4.12–3 .99(m,2H),3.81-3.73(m,1H),3.48(s,3H),3.34–3.16(m,3H),3.01–2.80(m,3H),1.97–1.75(m,2H).

Peak 2: LC-MS m/z=440.1[M+1] ⁺ .

¹ H NMR (400MHz, CDCl ₃ )δ8.58(d,1H),7.73(d,1H),7.67(dd,1H),7.48(d,1H),7.32(d,1H),7.28(d,1H),5.17(dt,1H),4.13–4.01(m,2H),3 .83-3.75(m,1H),3.48(s,3H),3.38(dd,1H),3.33–3.19(m,2H),3.07(dd,1H),3.02–2.87(m,2H),2.06–1.78(m,2H).

Examples 37 and 38: (S)-N-((S)-1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide and (S)-N-((R)-1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compounds 37 and 38)

(S)-N-((S)-1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazepane-2-carboxamide and(S)-N-((R)-1 -cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 37 and compound 38)

Step 1: 2-Bromo-5-(Bromomethyl)pyridine (37B)

2-bromo-5-(bromomethyl)pyridine(37B)

37A (5 g, 29.07 mmol) was dissolved in carbon tetrachloride (50 mL), and N-bromosuccinimide (5.43 g, 30.52 mmol) and azobisisobutyronitrile (1.19 g, 7.27 mmol) were added. The mixture was reacted at 90 °C for 2.5 h. After cooling to room temperature, the reaction solution was concentrated, and the residue was purified by silica gel column chromatography (PE:EA (v/v) = 10:1) to give the title compound 37B (4.42 g, 61%). LC-MS (ESI): m/z = 251.9 [M+H] ⁺ .

Step 2: 3-(6-bromopyridin-3-yl)-2-((diphenylmethylene)amino)propionitrile (37D)

3-(6-bromopyridin-3-yl)-2-((diphenylmethylene)amino)propanenitrile(37D)

37B (4.42 g, 17.54 mmol) and 37C (3.86 g, 17.54 mmol) were dissolved in dichloromethane (50 mL), and benzyltrimethylammonium chloride (0.33 g, 1.75 mmol) was added. Then, with vigorous stirring, an aqueous solution of sodium hydroxide (1.40 g, 35.08 mmol) (5 mL) was added, and the mixture was reacted overnight at room temperature. 100 mL of water was added, and the mixture was extracted with dichloromethane (30 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (PE:EA (v/v) = 10:1) to give the title compound 37D (5.20 g, 76%).

¹ H NMR (400MHz, CDCl ₃ )δ8.16(s,1H),7.64–7.57(m,2H),7.50–7.32(m,8H),6.96(q,J＝2.4Hz,2H) ,4.40(t,J＝6.5Hz,1H),3.17(d,J＝6.5Hz,2H).LC-MS(ESI):m/z＝390.0[M+H] ⁺ .

Step 3: 2-((diphenylmethylene)amino)-3-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)propionitrile (37E)

2-((diphenylmethylene)amino)-3-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)propanenitrile(37E)

37D (1.5 g, 3.84 mmol), 1A (1.06 g, 3.84 mmol), 1,1'-(diphenylphosphine)ferrocene]dichloropalladium (0.28 g, 0.38 mmol), and potassium carbonate (1.59 g, 11.52 mmol) were dissolved in a mixed solvent of dioxane (50 mL) and water (5 mL). The reaction was carried out at 90 °C for 5 h under nitrogen protection. After cooling to room temperature, 200 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic layers were combined and washed successively with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL). The mixture was dried over anhydrous sodium sulfate, and the residue was concentrated and purified by silica gel column chromatography (PE:EA (v/v) = 3:1) to give the title compound 37E (1.02 g, 57%). LC-MS (ESI): m/z = 459.1 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ )δ8.48(s,1H),7.74(s,1H),7.69–7.61(m,5H),7.49–7.42(m,4H),7.36(t,J＝7.5Hz,2H),7 .28–7.24(m,1H),6.98(d,J＝7.7Hz,2H),4.50–4.42(m,1H),3.47(s,3H),3.29–3.24(m,2H).

Step 4: 2-Amino-3-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)propionitrile (compound 37F)

2-amino-3-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)propanenitrile(compound 37F)

37E (1.02 g, 2.19 mmol) was dissolved in tetrahydrofuran (50 mL) and water (5 mL), and 2.5 mL of 1 M HCl aqueous solution was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature for 5 h. The reaction mixture was extracted with diethyl ether (15 mL x 3) and discarded. The aqueous layer was adjusted to pH 12 with 2 M NaOH aqueous solution and extracted with DCM (20 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated to obtain 580 mg of crude 37F, the title compound. This crude product was used directly in the next reaction without further purification. LC-MS (ESI): m/z = 295.1 [M+H] ⁺ .

Step 5: (2S)-2-((1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (compound 37G)

tert-butyl(2S)-2-((1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate(compound 37G)

INT-3 (0.36 g, 1.22 mmol) was dissolved in DMF (5 mL). Under nitrogen protection, HATU (0.6 g, 1.59 mmol) and N,N-diisopropylethylamine (0.47 g, 3.66 mmol) were added. After stirring at room temperature for 20 min, 37F (0.3 g, 1.22 mmol) was added, and the reaction was allowed to proceed for 3 h at room temperature. 30 mL of water was added to the reaction mixture, and the mixture was extracted with EA (15 mL x 5). The organic layers were combined, dried over anhydrous sodium sulfate, and the residue was concentrated. The residue was purified by silica gel column chromatography (DCM:MeOH (v/v) = 97:3) to give the title compound 37G (0.32 g, 51%). LC-MS (ESI): m/z = 522.2 [M+H] ⁺ .

Step 6: (S)-N-(1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (compound 37H)

(S)-N-(1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridin-3-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 37H)

37G (0.32 g, 0.61 mmol) was dissolved in anhydrous formic acid (3 mL) and reacted at 50 °C for 1 h. After cooling to room temperature, most of the solvent was removed by concentration. A saturated sodium bicarbonate solution (25 mL) was added to the residue, and the mixture was extracted multiple times with EA until no product was found in the aqueous layer. The organic layers were combined, dried over anhydrous sodium sulfate, and the residue was purified by silica gel column chromatography (DCM:MeOH (v/v) = 10:1) to give compound 37H (140 mg, 55%).

Compound 37H was isolated by chiral preparation via SFC to obtain peak 1 (63 mg, ee% = 98.22%, yield 24.8%, retention time: 2.120 min, designated as compound 37) and peak 2 (73 mg, ee% = 100%, yield: 28.7%, retention time: 2.689 min, designated as compound 38).

The purification conditions were as follows: ₍ Instrument name: MG II preparative SFC (SFC-14); Column: ChiralPak AD, 250×30mm ID, 10μm; Mobile phase: Phase A: _CO2 ; Phase B: Methanol (0.1% _NH3H2O ); Flow rate: 80mL/min; Column pressure: 100bar; Column temperature: 35℃; Absorption wavelength: 220nm; Cycle time: approximately 16.9min.)

Peak 1: LC-MS (ESI): m/z=422.3[M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD)δ8.55(s,1H),7.90–7.82(m,2H),7.79(m,2H),7.37–7.31(m,1H),5.17(d d,J＝9.0,6.8Hz,1H),4.11(dd,J＝8.6,3.6Hz,1H),4.03–3.95(m,1H),3.78(m, 1H),3.47(s,3H),3.34(m,1H),3.27–3.14(m,2H),2.90(m,1H),2.78(m,1H),2. 62(dd,J＝14.4,8.6Hz,1H),1.96–1.78(m,2H). Peak 2: LC-MS(ESI): m/z＝422.3[M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD)δ8.55(s,1H),7.90–7.82(m,2H),7.81–7.76(m,2H),7.36–7.31(m,1H),5.12(dd,J＝8.6,6.8Hz,1H),4.11(dd,J＝8.6,3.6 Hz,1H),4.05(m,1H),3.79(m,1H),3.47(s,3H),3.40–3.33(m,1H),3.29–3.20(m,2H),3.06–2.89(m,3H),2.02–1.83(m,2H).

Examples 39 and 40: (S)-N-((S)-1-cyano-2-(2-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyrimidinyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide and (S)-N-((R)-1-cyano-2-(2-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyrimidinyl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compounds 39 and 40)

(S)-N-((S)-1-cyano-2-(2-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyrimidin-5-yl)ethyl)-1,4-oxazepane-2-carboxamide and(S)-N-((R)-1 -cyano-2-(2-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyrimidin-5-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 39 and compound 40)

Step 1: (2-Chloroprene-5-yl)methanol (39B)

(2-chloropyrimidin-5-yl)methanol

39A (2 g, 10.72 mmol) was dissolved in anhydrous THF (20 mL) under nitrogen protection, and then the temperature was lowered to 0 °C. Diisobutylaluminum hydride (21.5 mL) was slowly added dropwise to the reaction solution. The mixture was stirred in an ice bath for 0.5 h. The reaction was monitored by TLC until complete. Saturated ammonium chloride (10 mL) was added to quench the reaction. The mixture was extracted with ethyl acetate (3 x 20 mL), and the combined organic phases were dried over anhydrous sodium sulfate. Column chromatography yielded product 39B (1.2 g, 77.4%) (PE:EA = 1:1). LC-MS (ESI): m/z = 145.1 [M+H] ⁺ .

Step 2: 5-(bromomethyl)-2-chloropyrimidine (39C)

5-(bromomethyl)-2-chloropyrimidine

39B (1 g, 6.92 mmol) was dissolved in DCM (20 ml) at room temperature. The reaction solution was cooled to 0 °C under nitrogen protection, and triphenylphosphine (1.82 g, 6.92 mmol) and carbon tetrabromide (2.29 g, 6.92 mmol) were added while maintaining this temperature. After stirring at 0 °C for 0.5 h, the reaction solution was heated to room temperature and stirred for another 1.5 h. TLC showed a small amount of substrate remaining. The reaction solution was evaporated to dryness and loaded onto a column (PE:EA = 5:1) to obtain 39C (1 g, 70%). LC-MS (ESI): m/z = 207.1 [M+H] ⁺ .

Step 3: 3-(2-chloropyrimidin-5-yl)-2-((diphenylmethylene)amino)propionitrile (39D)

3-(2-chloropyrimidin-5-yl)-2-((diphenylmethylene)amino)propanenitrile

39C (1 g 4.82 mmol), diphenylmethyleneaminoacetonitrile (1.06 g 4.82 mmol), and benzyltrimethylammonium chloride (0.18 g 0.96 mmol) were dissolved in DCM (30 ml) at room temperature. NaOH (0.8 ml 19 mol/L) was slowly added to the reaction solution, and the reaction solution was stirred overnight at room temperature. The reaction was quenched by adding 20 ml of water, extracted with DCM (20 ml x 3), and the organic phases were combined, dried over anhydrous sodium sulfate, and evaporated to dryness before column chromatography (PE:EA = 3:1) to give product 39D (1.1 g, 65.8%). LC-MS (ESI): m/z = 347.1 [M+H] ⁺ .

Step 4: 2-((diphenylmethylene)amino)-3-(2-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyrimidin-5-yl)propionitrile (39E)

2-((diphenylmethylene)amino)-3-(2-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyrimidin-5-yl)propanenitrile

Reactants 39D (1.1 g, 3.17 mmol) and 1A (0.96 g, 3.49 mmol) were dissolved in 1,4-dioxane (100 ml), followed by the addition of potassium carbonate (1.31 g, 9.51 mmol) and Pd(dppf) _Cl₂ (0.23, 0.32 mmol). The mixture was then subjected to nitrogen purging and reacted at 100°C. TLC and LC-MS showed a small amount of reactants remaining. The mixture was concentrated and evaporated to dryness, dissolved in DCM, filtered through diatomaceous earth, and the filtrate was evaporated to dryness and passed through a column. EA/PE = 0%–40%. The column chromatography yielded a pale yellow solid 39E (1.1 g, 75.5%). LC-MS (ESI): m/z = 460.2 [M+H] ^⁺ .

Step 5: 2-Amino-3-(2-(3-(2-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl]pyrimidin-5-yl)propionitrile (39F)

2-amino-3-(2-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyrimidin-5-yl)propanenitrile

Substrate 39E (1.1 g, 2.39 mmol) was dissolved in DCM (30 ml), and 1 M HCl (7.2 ml) was added. The mixture was stirred at room temperature for 3 h. TLC showed complete reaction of the starting material. The pH of the solution was adjusted to 8-10, and the mixture was extracted with EA (20 ml x 2). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness to obtain product 39F (0.6 g, 85%). LC-MS (ESI): m/z = 296.1 [M+H] ⁺ .

Step 6: (2S)-2-(1-cyano-2-(2-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyrimidin-5-yl)ethylcarbamoyl)-1,4-oxazacycloheptane-4-carboxylic acid tert-butyl ester (39G)

(2S)-tert-butyl 2-(1-cyano-2-(2-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyrimidin-5-yl)ethylcarbamoyl)-1,4-oxazepane-4-carboxylate

INT-3 (0.5 g, 2.03 mmol), HATU (0.85 g, 2.23 mmol), and DIPEA (0.7 ml) were dissolved in DCM (30 ml) at room temperature. After stirring for 10 min at room temperature, 39F (0.6 g, 2.03 mmol) was added, and the mixture was stirred for 1 h at room temperature. The reaction was monitored by TLC until complete. The reaction was quenched with water (10 ml), extracted with dichloromethane (15 ml × 2), and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (PE:EA = 1:1) to give product 39G (0.6 g, 56%). LC-MS (ESI): m/z = 523.2 [M+H] ⁺ .

Step 7: (2S)-N-(1-cyano-2-(2-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyrimidin-5-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (39H)

(2S)-N-(1-cyano-2-(2-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyrimidin-5-yl)ethyl)-1,4-oxazepane-2-carboxamide

39G (0.6g, 1.15mmol) was dissolved in anhydrous formic acid (20ml). After stirring at room temperature for 2h, TLC analysis showed that the substrate reaction was complete. The formic acid was removed at low temperature, and the pH was adjusted to 8-10 by adding saturated sodium bicarbonate solution. Extraction was performed by DCM (20ml x 2), and the product was dried over anhydrous sodium sulfate. The extract was then purified by column chromatography (DCM: _CH3OH = 10:1) to obtain compound 39H (0.15g, 31%). Compound 39H was chirally separated by SFC to obtain peak 1 (retention time: 1.528min, designated as compound 39) and peak 2 (retention time: 2.387min, designated as compound 40).

The separation method is as follows: (Instrument name: MG II preparer SFC (SFC-14; Column: ChiralPakAD, 250×30mm I.D., 10μm; Mobile phase: Phase A: CO2; Phase B: Isopropanol (0.1% NH3·H2O); Flow rate: 80mL/min; Column pressure: 100bar; Column temperature: 35℃; Absorption wavelength: 220nm; Cycle time: 3min.)

Peak 1: ¹ H NMR (400MHz, DMSO-d ₆ )δ8.79-8.78(m,1H),8.38-8.35(m,1H),7.43-7.38(m,3H),6.48-6.46(m,1H),5.03-4.99(m,1H),4.05-4.02(m,1H),3.90 -3.84(m,1H),3.76-3.70(m,1H),3.50(s,3H),3.25-3.09(m,4H),2.86-2.80(m,1H),2.70-2.58(m,2H),1.80-1.71(m,2H).

Peak 2: ¹ H NMR (400MHz, DMSO-d ₆ )δ8.79-8.78(m,1H),8.38-8.35(m,1H),7.43-7.38(m,3H),6.48-6.46(m,1H),5.03-4.99(m,1H),4.05-4.02(m,1H),3.90 -3.84(m,1H),3.76-3.70(m,1H),3.50(s,3H),3.25-3.09(m,4H),2.86-2.80(m,1H),2.70-2.58(m,2H),1.80-1.71(m,2H).

Examples 41 and 42: (S)-N-((S)-1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridazin-3-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide and (S)-N-((R)-1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridazin-3-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (Compounds 41 and 42)

(S)-N-((S)-1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridazin-3-yl)ethyl)-1,4-oxazepane-2-carboxamide and(S)-N-((R)-1 -cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridazin-3-yl)ethyl)-1,4-oxazepane-2-carboxamide(compound 41 and compound 42)

Step 1: 3-(bromomethyl)-6-chloropyridazine (41B)

3-(bromomethyl)-6-chloropyridazine

3-(methyl)-6-chloropyridazine (12.8 g, 100 mmol) was dissolved in carbon tetrachloride (300 mL), N-bromosuccinimide (17.8 g, 100 mmol) was added, followed by azobisisobutyronitrile (3.4 g, 20 mmol). The mixture was reacted overnight at 70 °C. After filtration and concentration, the filtrate was purified by silica gel column chromatography (PE:EA(v/v) = 1:10–1:5) to give the title compound (41B), (4 g, 20% yield). LCMS m/z = 207.46 [M+1] ⁺

Step 2: Ethyl 3-(6-chloropyridazine-3-yl)-2-((diphenylmethylene)amino)propionate (41C)

ethyl 3-(6-chloropyridazin-3-yl)-2-((diphenylmethylene)amino)propanoate

Ethyl 2-((diphenylmethylene)amino)acetate (6.18 g, 23.14 mmol), tetrabutylammonium bromide (9.32 g, 28.92 mmol), and potassium hydroxide (3.25 g, 2.24 mmol) were dissolved in a mixture of toluene (100 mL) and water (20 mL). 41B (4.0 g, 19.28 mmol) was added under ice bath conditions. The mixture was allowed to warm naturally to room temperature for 2 hours. Water (50 mL) was added, and the mixture was extracted with ethyl acetate (50 mL × 3), washed three times with water (50 mL × 3), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (PE:EA (v/v) = 1:10–1:5) to give the title compound (41C) (3.4 g, 45% yield). LCMS m/z = 394.12 [M+1] ⁺

Step 3: 3-(6-Chlorpyridazine-3-yl)-2-((diphenylmethylene)amino)propionamide (41D)

3-(6-chloropyridazin-3-yl)-2-((diphenylmethylene)amino)propanamide

41C (3.4 g, 8.7 mmol) was dissolved in ammonia-methanol solution (7 N, 50 mL), and the mixture was sealed in a tube (120 mL) and reacted overnight at 80 °C. After concentration, the mixture was purified by silica gel column chromatography (MeOH:DCM (v/v) = 1:10) to give the title compound 41D (1.8 g, yield 56%). LCMS m/z = 365.11 [M+1] ⁺

Step 4: 3-(6-Chlorpyridazin-3-yl)-2-((diphenylmethylene)amino)propionitrile (41E)

3-(6-chloropyridazin-3-yl)-2-((diphenylmethylene)amino)propanenitrile

41D (1.8 g, 4.93 mmol) was dissolved in dichloromethane (30 mL), and Burgess reagent (2.35 g, 9.86 mmol) was added. The mixture was reacted at room temperature for 2 hours, concentrated, and purified by silica gel column chromatography (PE:EA (v/v) = 1:5 to 5:5) to give the title compound 41E (1.4 g, 82% yield). LCMS m/z = 347.13 [M+1] ⁺

Step 5: 2-((diphenylmethylene)amino)-3-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridazin-3-yl)propionitrile (41F)

2-((diphenylmethylene)amino)-3-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridazin-3-yl)propanenitrile

41E (1.4 g, 4.04 mmol) was dissolved in dioxane (30 mL), 1A (2.22 g, 8.08 mmol), potassium carbonate (1.67 g, 12.12 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (590 mg, 0.81 mmol) were added, followed by water (6 mL). The mixture was reacted in a microwave reactor under nitrogen protection at 120 °C for 2 hours. After concentration, the mixture was purified by silica gel column chromatography (PE:EA (v/v) = 1:5 to 1:1) to give the title compound 41F (800 mg, yield 43%). LCMS m/z = 460.17 [M+1] ⁺

Step 6: 2-Amino-3-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridazin-3-yl)propionitrile (41G)

2-amino-3-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridazin-3-yl)propanenitrile

41F (800 mg, 1.74 mmol) was dissolved in dioxane (20 mL), and hydrochloric acid solution (0.5 N, 8 mL) was added. The reaction was carried out at room temperature for 0.5 h. The pH was adjusted to 7–8 with saturated sodium carbonate aqueous solution, and the mixture was extracted with dichloromethane (30 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (MeOH:DCM (v/v) = 1:100–1:10) to give the title compound 41G (300 mg, yield 58%). LCMS m/z = 296.11 [M+1] ⁺

Step 7: (2S)-2-((1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl]pyridazin-3-yl)ethyl)carbamoyl)-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (41H)

tert-butyl(2S)-2-((1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridazin-3-yl)ethyl)carbamoyl)-1,4-oxazepane-4-carboxylate

41G (300 mg, 1.02 mmol) was dissolved in DMF (20 mL), and HATU (390 mg, 1.02 mmol), DIPEA (260 mg, 2.02 mmol), and INT-3 (250 mg, 1.02 mmol) were added sequentially. The mixture was reacted at room temperature for 12 hours, followed by the addition of water (30 mL). The mixture was extracted with ethyl acetate (30 mL × 3), and the organic phases were combined. The organic phases were washed with water (30 mL × 2), then with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (MeOH:DCM (v/v) = 1:100–1:10) to give the title compound 41H (300 mg, yield 56%). LCMS m/z = 523.22 [M+1] ⁺

Step 8: (S)-N-((S)-1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridazin-3-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide and (S)-N-((R)-1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridazin-3-yl)ethyl)-1,4-oxazacycloheptane-2-carboxamide (compounds 41 and 42)

(S)-N-((S)-1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridazin-3-yl)ethyl)-1,4-oxazepane-2-carboxamide a nd(S)-N-((R)-1-cyano-2-(6-(3-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)pyridazin-3-yl)ethyl)-1,4-oxazepane-2-carboxamide

41H (300 mg, 0.57 mmol) was dissolved in acetonitrile (20 mL), p-toluenesulfonic acid (332 mg, 1.71 mmol) was added, and the mixture was reacted at 30 °C for 3 hours. The pH was adjusted to 7–8 with saturated sodium carbonate aqueous solution, and the mixture was extracted with dichloromethane (40 mL × 3). The organic phases were combined and washed with saturated sodium chloride aqueous solution (50 mL). The mixture was dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (DCM:MeOH (v/v) = 0.01:1–0.1:1) to obtain the crude compound.

150 mg of the crude compound was separated by chiral preparation, yielding two optical isomers: peak 1 (retention time: 2.088 min, 30 mg, ee = 99%, designated as compound 41) and peak 2 (retention time: 2.955 min, 30 mg, ee = 99%, designated as compound 42). Resolution conditions: Instrument: MG II preparative SFC (SFC-14); Column: Chiral Pak AD, 250 × 30 mm ID; Mobile phase: A: CO2, B: ethanol (0.05% _NH3H2O ); Gradient: B 40%; Flow rate: 70 mL/min; Back pressure: 100 bar; Column temperature: 38℃; Wavelength: 220 nm; Period: 13 _min ; Sample preparation: Compound 1 was dissolved in methanol to obtain 15 mg/mL; Injection: 1.0 mL/needle.

Peak 1: MS M/Z (ESI): m/z=423.17[M+1] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.73(d,1H),8.27(d,1H),8.08(d,1H),8.00–7.90(m,1H),7.78(d,1H),7.50(d,1H),5.41–5.32(m,1H),4 .02–3.84(m,2H),3.78–3.49(m,3H),3.44(s,3H),3.18–3.07(m,1H),2.90–2.53(m,4H),1.82–1.63(m,2H).

Peak 2: MS M/Z (ESI): m/z=423.17[M+1] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ )δ8.75(d,1H),8.27(d,1H),8.07(d,1H),7.98–7.86(m,1H),7.78(d,1H),7.50(d,1H),5.62–5.19(m,1H),4.05–3.97(m,1H),3.92–3.82 (m,1H),3.77–3.66(m,1H),3.66–3.51(m,2H),3.43(s,3H),3.09–3.00(m,1H),2.86–2.70(m,1H),2.69–2.51(m,3H),1.80–1.63(m,2H).

Example 43: N-((S)-1-cyano-2-(3-fluoro-4'-(pentafluoro-16-thio)-[1,1'-biphenyl]-4-yl)ethyl)-6-methoxy-1,4-oxazacycloheptane-2-carboxamide (Compound 43)

N-((S)-1-cyano-2-(3-fluoro-4'-(pentafluoro-l6-sulfanyl)-[1,1'-biphenyl]-4-yl)ethyl)-6-methoxy-1,4-oxazepane-2-carboxamide(compound 43)

Step 1: (2S,6R)-2-(((benzyloxy)methyl)-6-methoxy-4-toluenesulfonyl-1,4-oxazacycloheptane (43B)

(2S,6R)-2-((benzyloxy)methyl)-6-methoxy-4-tosyl-1,4-oxazepane(43B)

43A (0.5 g, 1.28 mmol, preparation method reference: Eur. J. Org. Chem. 2007, 2107–2113 (DOI: 10.1002/ejoc.200700011)) was dissolved in tetrahydrofuran (20 mL). Sodium hydride (0.26 g, 6.4 mmol) was added at 0 °C, and the mixture was stirred for 30 min. Then, iodomethane (0.91 g, 6.4 mmol) was added, and the mixture was reacted at room temperature for 3 h. Water (30 mL) was added at 0 °C, and the mixture was extracted with EA (20 mL x 3). The extract was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a colorless oily substance 43B (0.4 g, yield 77%).

Step 2: (2S,6R)-2-((benzyloxy)methyl)-6-methoxy-1,4-oxazacycloheptane (43C)

(2S,6R)-2-((benzyloxy)methyl)-6-methoxy-1,4-oxazepane(43C)

43B (0.3 g, 0.74 mmol) was dissolved in methanol (10 mL), and magnesium filings (2.36 g, 97.08 mmol) were added. The mixture was sonicated at 50 °C for 2 h, then reacted at room temperature for 16 h. The solution was filtered and concentrated to obtain a white oily substance, 43C (0.18 g, 96% yield), which was then directly proceeded to the next step. LC-MS (ESI): m/z = 252.3 [M+1] ⁺

Step 3: (2S,6R)-2-((benzyloxy)methyl)-6-methoxy-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (43D)

(2S,6R)-tert-butyl 2-((benzyloxy)methyl)-6-methoxy-1,4-oxazepane-4-carboxylate(43D)

43C (0.18 g, 0.72 mmol) was dissolved in DCM (10 mL), and triethylamine (0.087 g, 0.85 mmol) and TBSCl (0.37 g, 1.71 mmol) were added sequentially. Under nitrogen protection, the mixture was reacted at room temperature for 2 hours, concentrated to dryness, and purified by column chromatography (PE:EA = 10:1-4:1) to obtain a colorless oily substance 43D (0.12 g, yield 47%).

Step 4: (2S,6R)-2-(hydroxymethyl)-6-methoxy-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (43E)

(2S,6R)-tert-butyl 2-(hydroxymethyl)-6-methoxy-1,4-oxazepane-4-carboxylate (43E): 43D (0.12 g, 0.34 mmol) was dissolved in methanol (10 mL), and palladium on carbon (0.1 g, 10%) was added. The mixture was reacted under a hydrogen atmosphere for 24 hours, filtered, concentrated to dryness, and a colorless oily substance 43E (0.08 g, 86% yield) was obtained.

Step 5: (2S,6R)-4-(tert-Butoxycarbonyl)-6-methoxy-1,4-oxazacycloheptane-2-carboxylic acid (43F)

(2S,6R)-4-(tert-butoxycarbonyl)-6-methoxy-1,4-oxazepane-2-carboxylicacid (43F) was prepared by dissolving 43E (0.095 g, 0.3 mmol) in acetone (7 mL), adding saturated sodium bicarbonate (3 mL), sodium bromide (0.025 g, 0.24 mmol), and TEMPO (0.004 g, 0.024 mmol). Trichloroisocyanuric acid (0.25 g, 1.06 mmol) was added at 0 °C, and the reaction was allowed to proceed for 16 hours. The pH was adjusted to 5-6 with dilute hydrochloric acid, and the mixture was extracted with water (30 mL) and dichloromethane (20 mL x 3). The extract was washed with water (30 mL) and saturated sodium chloride (30 mL), dried over anhydrous sodium sulfate, and concentrated to dryness to obtain a yellow oily substance, 43F (0.046 g, 44% yield).

Step 6: 2-(((S)-1-cyano-2-(3-fluoro-4'-(pentafluoro-16-thio)-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-6-methoxy-1,4-oxazacycloheptan-4-carboxylic acid tert-butyl ester (43G)

tert-butyl-2-(((S)-1-cyano-2-(3-fluoro-4'-(pentafluoro-l6-sulfanyl)-[1,1'-biphenyl]-4-yl)ethyl)carbamoyl)-6-methoxy-1,4-oxazepane-4-carboxylate

Compounds 8B (400 mg, 1.1 mmol), 43F (300 mg, 1.1 mmol), HATU (458 mg, 1.65 mmol), and DIEA (0.54 mL, 3.3 mmol) were mixed and dissolved in DMF, and stirred overnight at room temperature. The reaction was monitored by LC-MS until complete. Water was added, followed by extraction with EA. The organic phase was dried and concentrated, and separated by column chromatography (DCM:MeOH = 10:1) to give the title compound 43G (512 mg, 75%). LC-MS (ESI): m/z = 624.2 [M+H] ⁺ .

Step 7: N-((S)-1-cyano-2-(3-fluoro-4'-(pentafluoro-16-thio)-[1,1'-biphenyl]-4-yl)ethyl)-6-methoxy-1,4-oxazacycloheptane-2-carboxamide (Compound 43)

N-((S)-1-cyano-2-(3-fluoro-4'-(pentafluoro-l6-sulfanyl)-[1,1'-biphenyl]-4-yl)ethyl)-6-methoxy-1,4-oxazepane-2-carboxamide

Compound 43 (512 mg, 0.82 mmol) was dissolved in 40 mL of acetonitrile, and p-toluenesulfonic acid (706 mg, 2.46 mmol) was added. The mixture was heated to 40 °C and reacted for 2 hours. The reaction was monitored by LC-MS until complete. The reaction solution was concentrated, and EA and saturated sodium bicarbonate aqueous solution were added. The mixture was extracted, separated, and the organic phase was dried and concentrated. The organic phase was then separated by column chromatography (DCM:MeOH = 10:1) to give the title compound 43 (75 mg, 17%). LC-MS (ESI): m/z = 524.2 [M+H] ⁺ .

¹ HNMR (400MHz, DMSO-d ₆ ): 8.74(d,1H),7.92-8.00(m,4H),7.48-7.66(m,3H),5.03-5.09(m,1H),3.97-4.01(m,2H) ,3.56-3.59(m,1H),3.28-3.30(m,1H),3.24-3.25(m,6H),2.78-2.89(m,3H),1.91(s,1H).

Biological testing

1. In vitro DPP1 enzyme activity detection experiment

Recombinant human DPP1 enzyme (R&D Systems, Cat. No. 1071-CY) at a final concentration of 100 μg/mL was mixed with recombinant human cathepsin L (R&D System, Cat. No. 952-CY) at a final concentration of 20 μg/mL and incubated at room temperature for 1 hour to activate the DPP1 enzyme. The activated DPP1 enzyme was diluted 100-fold, and 5 μL of different concentrations of the compound and 5 μL of diluted DPP1 enzyme were added to 384-well plates, incubated at room temperature for 30 minutes. Then, 10 μL of 20 μM substrate Gly-Arg-AMC (bachem, Cat. No. I-1215) was added, and incubation continued at room temperature for 60 minutes. Fluorescence intensity was detected using a microplate reader, with excitation at 380 nm and emission at 460 nm. The _IC50 value was calculated using the DosResp function in Origin 2019 software.

Test results: The compounds of this invention exhibited inhibitory activity against the DPP1 receptor. The IC50 values of the compounds in the examples against the DPP1 receptor were less than 100 nM. The test results for some of the examples are shown in Table 1.

Table 1 DPP1 inhibitory activity

Compound numbering <![CDATA[IC₅₀/nM]]> Compound 1 1.6 Compound 10 6.7 Compound 11 239 Compound 12 twenty one Compound 13 71 Compound 14 12.9 Compound 15 33 Compound 17 7.4 Compound 23 0.5 Compound 29 4.9 Compound 35 4.0 Compound 43 0.3

Conclusion: The compounds of this invention exhibit high inhibitory activity against the DPP1 receptor.

2. Rat pharmacokinetic test

1.1 Experimental animals: Male SD rats, approximately 220g, 6-8 weeks old, 6 rats/compound. Purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

1.2 Experimental Design: On the day of the experiment, 6 SD rats were randomly divided into groups according to their body weight. They were fasted for 12-14 hours before drug administration but allowed free access to water. They were fed 4 hours after drug administration.

Table 2 Drug Administration Information

Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Gavage administration solvent: 0.5% MC; Control compound INS1007, namely compound 2 in patent WO2015110826A1, was prepared according to the patented method.

Blood samples of 0.1 ml were collected via the orbital cavity before and after isoflurane anesthesia, placed in EDTAK2 centrifuge tubes, and centrifuged at 5000 rpm for 10 min at 4°C to collect plasma. Blood collection time points for the intravenous group: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 24 h; blood collection time points for the gavage group: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 24 h. All samples were stored at -80°C before analysis.

Table 3. Pharmacokinetic parameters of the tested compounds in rat plasma.

Conclusion: The compounds of this invention have good bioavailability and pharmacokinetic characteristics.

3. Repeated oral administration toxicity test in rats over 14 days

SD rats were randomly divided into three groups according to body weight: a solvent control group (0.5% MC), an INS1007 (30, 100, 300 mg/kg) group, and a compound (30, 100, 300 mg/kg) group, with 16 rats in each drug administration group and 10 rats in each solvent control group. The rats were half male and half female. The appropriate concentration of drug or solvent was administered orally by gavage daily for 14 consecutive days, followed by a 7-day recovery period. During the drug administration period, general symptoms, body weight, and food intake were monitored in each group. At the end of the drug administration period and the end of the recovery period, hematological, serum biochemical, and gross anatomical examinations were performed in each group.

Conclusion: At the same dosage, the compounds of this invention are less toxic than INS1007 and have higher safety.

Claims (13)

1. A compound of formula (IV), or a pharmaceutically acceptable salt thereof: in, Rc is H; _R1 , _R2 and _R3 are each independently selected from H, deuterium, F, Cl, Br, methyl, ethyl, methoxy or ethoxy, wherein the methyl, ethyl, methoxy or ethoxy group is optionally substituted by 1 to 3 groups selected from F, Cl, Br, cyano, hydroxyl and _NH2 ; Z is CH or N; Ring G is a benzene ring optionally substituted with 1-2 _RGs or Each R _G is independently selected from F, Cl, Br, I, methyl, ethyl, propyl, _SF5 and CN, wherein the methyl, ethyl or propyl group is optionally further substituted by 1 to 3 groups selected from deuterium, F, Cl, Br and I. 2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein Z is CH. 3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the ring G is optionally substituted with 1-2 _RGs . 4. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein each R _G is independently selected from methyl, ethyl, propyl, wherein the methyl, ethyl, or propyl group is optionally further substituted by 1-3 groups selected from deuterium, F, Cl, Br, and I. 5. The compound of claim 4, or a pharmaceutically acceptable salt thereof, wherein each _RG is independently selected from methyl, ethyl, or propyl. 6. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein the compound optionally substituted with 1-2 _RGs is selected from one of the following structures: 7. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from one of the following structures: 8. A compound, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from one of the following structures: 9. A compound, or a pharmaceutically acceptable salt thereof, characterized in that the compound is selected from the following structures: 10. A compound, or a pharmaceutically acceptable salt thereof, characterized in that the compound is selected from the following structures: 11. A pharmaceutical composition comprising the compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and/or excipient. 12. Use of the compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, or the composition of claim 11, in the preparation of a medicament for treating dipeptidyl peptidase 1-mediated diseases selected from airway obstructive diseases. 13. Use of the compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, or the composition of claim 11, in the preparation of a medicament for treating dipeptidyl peptidase 1-mediated diseases selected from bronchiectasis, cystic fibrosis, asthma, emphysema, and chronic obstructive pulmonary disease.