EA040139B1 - AZITIDINE DERIVATIVE - Google Patents

Description

Link to related applications

This Application Claims Priority Under Patent Applications

of China with serial Nos. 201710112350.0 and 201710805883.7 filed with the Chinese National Intellectual Property Office on February 28, 2017 and September 8, 2017, respectively, which are incorporated herein by reference in their entirety.

The field of technology to which the present invention relates

The present application relates to a class of compounds or a pharmaceutically acceptable salt thereof for the treatment of multiple myeloma, as well as their use in the preparation of a medicament for the treatment of a disease associated with multiple myeloma.

Background of the Invention

Multiple myeloma (MM) is a malignant proliferative disease of plasma cells, which is characterized by abnormal proliferation of clonal plasma cells in the bone marrow, impaired hematopoietic function, stimulation of the bone, which leads to osteolytic bone lesions and the presence of a monoclonal immunoglobulin or its fragment (protein M), which found in serum and/or urine. Its clinical manifestations are bone pain, anemia, hypercalcemia, kidney failure, infection, bleeding, etc. Bortezomib is a reversible proteasome inhibitor that achieves the goal of treating multiple myeloma by promoting apoptosis of myeloma cells. However, during long-term treatment, some patients with multiple myeloma develop resistance to bortezomib. Therefore, there is still a need for a new and safe drug for the treatment of multiple myeloma.

The essence of the present invention

In one aspect, the invention relates to a compound of formula (I), a pharmaceutically acceptable salt or stereoisomer thereof

where ring A is selected from the group consisting of C _3-6 cycloalkyl, phenyl and 5-10 membered heteroaryl, where the prefix hetero in 5-10 membered heteroaryl is each independently selected from the group consisting of -O-, -S -, -NH- and N; and the number of heteroatoms or a group containing a heteroatom is independently selected from 1, 2 or 3;

n is selected from 0, 1,2 or 3;

R ₁ is each independently selected from the group consisting of halogen, OH, NH ₂ , CN, C _1-3 alkyl, C _1-3 alkoxy and phenyl, where C _1-3 alkyl, C _1-3 alkoxy or phenyl are each unsubstituted or substituted with 1, 2 or 3 R;

R ₃ is H;

R2 is selected from the group consisting of H, halogen, OH, NH ₂ , CN, C _1-6 alkyl, C _1-6 alkoxy, C _1-3 alkylO-C1.3 alkyl-, C _3-6 cycloalkyl-(CH ₂ ) _1-3 - and phenyl-(CH ₂ ) _1-3 -, where C _1-6 alkyl, C _1-6 alkoxy, C _1-3 alkyl-OC _1-3 alkyl-, C _3-6 cycloalkyl- (CH ₂ ) _1-3 - or phenyl-(CH ₂ ) _1-3 - each unsubstituted or substituted with 1, 2 or 3 R; or

R2 and R3 together with the carbon atom to which they are attached form a 3-6 membered cycloalkyl;

R4 is selected from the group consisting of C _1-6 alkyl and C _3-6 cycloalkyl-(CH ₂ ) _1-3 -;

R5 is selected from the group consisting of H and C _1-3 alkyl;

each R is independently selected from the group consisting of F, Cl, Br, I, OH, Me, NH ₂ , NH(CH ₃ ) and N(CH3) 2 .

In one embodiment, n is selected from 0, 1, or 2. In yet another embodiment, R ₁ is each independently selected from the group consisting of halogen, OH, NH ₂ , CN, C _1-3 alkoxy, phenyl, and C _1-3 alkyl , unsubstituted or substituted with 1, 2 or 3 F.

In yet another embodiment, R1 is each independently selected from the group consisting of F, Cl, n

CN, CF3 and \=/·

In yet another embodiment, ring A is selected from the group consisting of cyclopropyl, phenyl, 5-membered heteroaryl, 6-membered heteroaryl, and 9-membered heteroaryl.

In yet another embodiment, Ring A is selected from the group consisting of cyclopropyl, phenyl, pyridyl, pyrimidinyl, pyrazinyl, and thiazolyl.

(RO-^-y η 7 d 1

In another embodiment, the structural unit in the compound of formula (I) is selected from the group consisting of

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implementation structural unit

In yet another embodiment, is selected from the group consisting of in a compound of formula (I)

In yet another embodiment

R3 is H;

R2 is selected from the group consisting of H, C1-6 alkyl, C _1-3 alkoxy, C _1-3 alkyl-OC _1-3 alkyl-, C3-6 cycloalkyl-CH ₂ - and phenyl-CH ₂ -, where C _1-6 alkyl, C _1-3 alkoxy, C _1-3 alkyl-OC _1-3 alkyl-, C _3-6 cycloalkyl-CH ₂ - or phenyl-CH ₂ - each unsubstituted or substituted with 1, 2 or 3 substituents, independently selected from the group consisting of F, Cl, Br, I and hydroxyl.

In another embodiment, R3 is H and R2 is selected from the group consisting of H, Me,

In yet another embodiment, R ₄ is selected from the group consisting of C ₃ - ₄ alkyl and C ₃ - ₄ cycloalkyl-CH ₂ -.

In yet another embodiment, R4 is selected from !

In yet another embodiment, R5 is selected from the group consisting of H and Me.

In yet another embodiment, each R is independently selected from the group consisting of F and OH.

In one embodiment, the compound of formula (I) is selected from the group consisting of a compound of formula (II), a compound of formula (III), a compound of formula (IV), a compound of formula (V), a compound of formula (VI), a compound of formula (I- a), compounds of formula (I-b), compounds of formula (II-a), compounds of formula (II-b), compounds of formula (III-a), compounds of formula (III-b), compounds of formula (IV-a), compounds formula (IV-b), compounds of formula (V-a), compounds of formula (V-b), compounds of formula (VI-a) and compounds of formula (VI-b)

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(Vl-b) where ring C is selected from the group consisting of cyclopropyl, 5-membered heteroaryl and 6-membered heteroaryl.

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In another embodiment, the structural unit

in formula (IV-a), formula (IV-b), formula (VI-a) or formula (VI-b) is selected from the group consisting of

In another embodiment, the compound of formula (I) is selected from the group consisting of

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or a pharmaceutically acceptable salt or stereoisomer thereof.

In one embodiment, the compound of formula (I) is selected from the group consisting of

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or a pharmaceutically acceptable salt thereof.

In one aspect of the invention, there is provided a pharmaceutical composition comprising a compound of the present invention, a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier, excipient, or adjuvant.

In one aspect of the invention, there is provided a method for the prevention or treatment of multiple myeloma, the method comprising administering to a subject in need thereof a compound of the present invention, a pharmaceutically acceptable salt or stereoisomer thereof.

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Detailed description of the present invention

The present invention relates to a compound of formula (I), a pharmaceutically acceptable salt, tautomer, stereoisomer or geometric isomer thereof.

where ring A is selected from the group consisting of C _3-6 cycloalkyl, phenyl and 5-10 membered heteroaryl;

n is selected from 0, 1,2 or 3;

R ₁ is each independently selected from the group consisting of halogen, OH, NH ₂ , CN, C _1-3 alkyl, C _1-3 heteroalkyl and phenyl, where C _1-3 alkyl, C _1-3 heteroalkyl or phenyl are each optionally substituted 1, 2 or 3 R;

R ₂ and R ₃ are each independently selected from the group consisting of H, halogen, OH, NH ₂ , CN, C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl, C _3-6 cycloalkyl-(CH ₂ ) _1-3 - and phenyl-(CH ₂ ) _1-3 -, where C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl, C _3-6 cycloalkyl-(CH ₂ ) _1-3 - or phenyl-(CH ₂ ) _1-3 - each is optionally substituted with 1, 2 or 3 R; or

R ₂ and R ₃ together with the carbon atom to which they are attached form a 3-6 membered ring;

R4 is selected from the group consisting of C _1-6 alkyl and C _3-6 cycloalkyl-(CH ₂ ) _1-3 -, where C _1-6 alkyl or C _3-6 cycloalkyl-(CH ₂ ) _1-3 is each optionally substituted with 1, 2 or 3 R;

R5 is selected from the group consisting of H and C _1-3 alkyl optionally substituted with 1, 2 or 3 R;

each R is independently selected from the group consisting of F, Cl, Br, I, OH, Me, NH ₂ , NH(CH ₃ ) and n(CH3) 2 ;

where the prefix hetero in C _1-3 heteroalkyl, C _1-6 heteroalkyl and 5-10-membered heteroaryl is each independently selected from the group consisting of -O-, -S-, -NH- and N; in any of the above cases, the number of heteroatoms or groups containing a heteroatom is independently selected from 1, 2 or 3.

According to some embodiments of the present application, n is selected from 0, 1, or 2.

According to some embodiments of the present application, the hetero prefix in C _1-3 heteroalkyl and C _1-6 heteroalkyl is each independently selected from the group consisting of -O-, -S- and -NH-, and the hetero prefix in 5-10 membered heteroaryl selected from the group consisting of N, -O- and -S-. In some embodiments of the present application, the hetero prefix in C _1-3 heteroalkyl and C _1-6 heteroalkyl is -O- and the hetero prefix in 5-10 membered heteroaryl is selected from the group consisting of N and -S-.

According to some embodiments of the present application, R ₁ is each independently selected from the group consisting of halogen, OH, NH ₂ , CN, C _1-3 alkyl, C _1-3 alkoxy and phenyl, where C _1-3 alkyl, C _1-3 alkoxy or phenyl are each optionally substituted with 1, 2 or 3 R; preferably R ₁ is each independently selected from the group consisting of halogen, OH, NH ₂ , CN, C _1-3 alkoxy, phenyl and C _1-3 alkyl optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of F, Cl, Br and I; more preferably R1 is each independently selected from the group consisting of halo, OH, NH ₂ , CN, C _1-3 alkoxy, phenyl and C _1-3 alkyl optionally substituted with 1, 2 or 3 F; even more preferably R1 is each independently selected from the group consisting of halo, CN, phenyl and C _1-3 alkyl optionally substituted with 3 F; and most preferably R ₁ is each independently selected from the group consisting of F, Cl CN, phenyl and methyl optionally substituted with 3 F.

According to some embodiments of the present application, R1 is each independently selected from . _n ........ ... .. Ah .., oh. H , a group consisting of F, Cl, Br, I, OH, NH ₂ , CN, Me, and where Me, ' ^{or 4=7} is optionally substituted with 1, 2 or 3 R.

According to some preferred embodiments of the present application, R1 each 0 0 is independently selected from the group consisting of F, Cl, Br, I, CN, Me and '—'> where Me or '—' is optionally substituted with 1, 2 or 3 substituents, independently selected from the group consisting of F, Cl, Br and I; preferably R1 is each independently selected from the group consisting of F, Cl, Br, I, CN, ^= ⁷ and Me optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of F, Cl, Br and I ; even more preferably R1 is each independently selected from the group consisting of F, Cl, Br, I,

CN, \=/ and Me optionally substituted with 1, 2 or 3 F; and most preferably R1 each independently

- 10 040139 Simo is selected from the group consisting of F, Cl, Br, I, CN,

and Me optionally substituted with 3 F.

According to some preferred embodiments of the present application, R1 is each independently selected from the group consisting of F, Cl, Br, I, OH, NH2, CN, Me, CF ₃ , > and preferably R ₁ is each independently selected from the group consisting of F, Cl, CN,

and CF3.

According to some embodiments of the present application, ring A is selected from the group consisting of phenyl and 5-10 membered heteroaryl.

In some embodiments of the present application, ring A is selected from the group consisting of cyclopropyl, phenyl, 5-membered heteroaryl, 6-membered heteroaryl, 7-membered heteroaryl, 8-membered heteroaryl, 9-membered heteroaryl, and 10-membered heteroaryl; preferably ring A is selected from the group consisting of cyclopropyl, phenyl, 5-membered heteroaryl, 6-membered heteroaryl and 9-membered heteroaryl; and more preferably ring A is selected from the group consisting of cyclopropyl, phenyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, 1,3,4 oxadiazolyl, thiazolyl, isothiazolyl, thienyl, pyrazolyl, imidazolyl and 1H-indazolyl.

According to some preferred embodiments of the present application, ring A is selected from the group consisting of cyclopropyl, phenyl, pyridyl, pyrimidinyl, thiazolyl and pyridazinyl, and more preferably ring A is selected from the group consisting of phenyl, pyridyl, pyrimidinyl, thiazolyl and pyridazinyl.

According to some preferred embodiments of the present application, ring A is selected from the group consisting of phenyl, pyridyl, pyrimidinyl, pyrazinyl, 1,3,4-oxadiazolyl, thiazolyl, imidazolyl, and 1H-indazolyl; preferably ring A is selected from the group consisting of phenyl, pyridyl and pyrimidinyl, and more preferably ring A is selected from phenyl.

According to some embodiments of the present application, the structural unit in the compound of formula (I) is selected from the group consisting of

According to some preferred embodiments of the present application, the structural (Ri)^^ η 7 l ) unit ⁴ is selected from the group consisting of

unit ⁿ b } _k ,. selected from the group consisting of

According to some preferred embodiments of the present application, the structural

selected from the group consisting of

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tour unit

According to some specific embodiments of the present application, the structural unit is selected from the group consisting of selected from

According to some more preferred embodiments of the present application, the structure

selected from the group consisting of even more preferably a structural unit

and most preferably the structural unit is selected from the group consisting of

According to some more preferred embodiments of the present application, the structural unit

selected from the group consisting of

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According to some specific embodiments of the present application, the structural unit

selected from the group consisting of

According to some preferred specific embodiments of the present application, ^p ( A ) is selected from the group consisting of a structural unit

selected from the group consisting of

More preferably a structural unit

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and even more preferably the structural unit is selected from the group consisting of

According to some preferred specific embodiments of the present application, ^η ( A ) structural unit is selected from the group consisting of

According to some embodiments of the present application, R2 and R3 are each independently selected from the group consisting of H, halo, OH, NH2, CN, C _1-6 alkyl, C _1-3 alkoxy, C _1-3 alkyl-OC ₁ _ ₃ alkyl -, C _3-6 cycloalkyl-CH ₂ - and phenyl-CH ₂ -, where C _1-6 alkyl, C1-3 alkoxy, C1-3 alkyl-OC ₁ _ ₃ alkyl-, C _3-6 cycloalkyl-CH ₂ - or phenyl-CH ₂ - each is optionally substituted with 1, 2 or 3 R; or R2 and R3 together with the carbon atom to which they are attached form a 3-6 membered cycloalkyl; preferably R 2 and R ₃ are each independently selected from the group consisting of H, C _1-6 alkyl, C 1-3 alkoxy, C 1-3 alkyl-OC ₁ _ ₃ alkyl-, C _3-6 cycloalkyl-CH ₂ - and phenyl- CH ₂ -, where C _1-6 alkyl, C 1-3 alkoxy, C 1-3 alkyl-OC ₁ _ ₃ alkyl-, C _3-6 cycloalkyl-CH ₂ - or phenyl-CH ₂ - each is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of F, Cl, Br, I and hydroxyl; more preferably R2 and _R3 are each independently selected from the group consisting of H _{, C1-6} alkyl, C1-3 alkoxy, _C3-6 cycloalkyl- _CH2- and phenyl- _CH2- ; even more preferably R2 and _R3 are _each independently selected from the group consisting of H, _C1-6 alkyl, _C3-6 cycloalkyl- _CH2- and phenyl- _CH2- ; and most preferably R2 and _R3 are each independently selected from the group consisting of H, Me,

According to some preferred embodiments of the present application, R ₃ is H and R 2 is selected from the group consisting of H, halo, OH, NH2, CN, _C1-6 alkyl, C1-3 alkoxy, C1-3 alkyl-OC ₁ _ ₃ alkyl -, C ₃ - ₆ cycloalkyl-CH ₂ - and phenyl-CH ₂ -, where C1- ₆ alkyl, C1-3 alkoxy, C1-3 alkyl-OC ₁ _ ₃ alkyl-, C ₃ - ₆ cycloalkyl-CH ₂ - or phenyl-CH ₂ - each is optionally substituted with 1, 2 or 3 R; preferably the aforementioned R ₃ is H and R 2 is selected from the group consisting of H, C _1-6 alkyl, C 1-3 alkoxy, C 1-3 alkyl-OC ₁ _ ₃ alkyl-, C ₃ - ₆ cycloalkyl-CH ₂ - and phenyl -CH ₂ -, where _C1-6 alkyl, C1-3 alkoxy, C1-3 alkyl-OC ₁ _ ₃ alkyl-, C ₃ - ₆ cycloalkyl-CH ₂ - or phenyl-CH ₂ - each is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of F, Cl, Br, I and hydroxyl; more preferably the aforementioned R ₃ is H and R 2 is selected from the group consisting of H, C _1-6 alkyl, C 1-3 alkoxy, C ₃ - ₆ cycloalkyl-CH ₂ - and phenyl-CH ₂ -; more preferably R ₃ is H

- 14 040139 and R2 is selected from the group consisting of H, C _1-6 alkyl, C _3-6 cycloalkyl-CH2- and phenyl-CH2-; and most preferably R ₃ is H and R 2 is selected from the group consisting of H, Me,

According to some embodiments of the present application, branes are from the group consisting of H, F, Cl, Br, I, OH,

, where Me, Et, or

R.

According to some embodiments of the present application, py, consisting of H, Me, is H.

R2 and R ₃ each independently

NH2, CN, Me, Et, optionally substituted 1, 2

R2 and R ₃ each independently

brane from the group consisting of H, Me,

According to some specific embodiments of the present application, R2 is selected from the group

you' and R ₃ represent According to some embodiments of the present application, R ₂ and R ₃ together with the carbon atom to which they are attached, form a 3-6 membered cycloalkyl.

According to some embodiments of the present application, the structural unit ^R ^ ^R3 is

According to some embodiments of the present application, R4 is selected from the group consisting of C1-6 alkyl and C ₃ - ₆ cycloalkyl-CH ₂ -, where C 1-6 alkyl or C ₃ - ₆ cycloalkyl-CH ₂ - each is optionally substituted with 1, 2 or 3R; preferably R4 is selected from the group consisting of C1-6 alkyl and C ₃ - ₆ cycloalkylCH ₂ -; more preferably R4 is selected from the group consisting of C1-4 alkyl and C ₃ - ₆ cycloalkyl-CH ₂ -; even more preferably R4 is selected from the group consisting of C ₃ - ₄ alkyl and C ₃ - ₄ cycloalkyl-CH ₂ -; and most preferably R4 is selected from the group consisting of C ₄ alkyl and C ₄ cycloalkyl-CH ₂ -.

According to some embodiments of the present application, R4 is selected from the group consisting of ^{1 and 1} , where I or - each is optionally substituted with 1, 2 or 3 R; preferably R4 is selected from the group consisting of ' and '; and more preferably R4 is '

In some embodiments of the present application, R5 is selected from the group consisting of H and C _1-3 alkyl; preferably R5 is selected from the group consisting of H, Me and Et; more preferably R ₅ is selected from the group consisting of H and Me; and most preferably R ₅ is H.

In some embodiments of the present application, each R is independently selected from the group consisting of F, C1, Br, I, OH, Me, and NH2; preferably each R is independently selected from the group consisting of F, C1, Br, OH and Me; and more preferably each R is independently selected from the group consisting of F and OH.

It should be understood that the above variables n, R, R1, R ₂ , R ₃ , R4, R5 and ring A according to the present application may be combined in any manner to form a plurality of embodiments η 7 e) ; and the aforementioned variables R, R ₂ , R ₃ , R4, R5 and structural unit can also be combined in any way to form a variety of embodiments.

In some embodiments of the present application, the compound of formula (I) is selected from the group consisting of

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or a pharmaceutically acceptable salt, tautomer, stereoisomer or geometric isomer thereof, wherein R1, R2, _R3 , R4 and R5 are as defined above.

The present application also relates to a compound of formula (II), a pharmaceutically acceptable salt, tautomer, stereoisomer or geometric isomer thereof.

where the ring A, n, R _b R2 and R5 are defined above.

ⁿ 7 l)

In some embodiments, the structural unit is as defined above.

The present application also relates to a compound of formula (III) or a compound of formula (IV), its pharmaceutically acceptable salt, tautomer, stereoisomer or geometric isomer

where n, R1, R2 and R5 are defined above; ring C is selected from the group consisting of cyclopropyl, 5-membered heteroaryl and 6-membered heteroaryl.

The present application also relates to a compound of formula (V) or a compound of formula (VI), its pharmaceutically acceptable salt, tautomer, stereoisomer or geometric isomer

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where n, R1, R2 and ring C are defined above.

According to some embodiments of the present application, ring C in the compound of formula (IV) or compound of formula (VI) is selected from the group consisting of cyclopropyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, 1,3,4-oxadiazolyl, thiazolyl, isothiazolyl, thienyl, pyrazolyl and imidazolyl; preferably ring C is selected from the group consisting of cyclopropyl, pyridyl, pyrimidinyl, thiazolyl and pyridazinyl; and more preferably ring C is selected from the group consisting of pyridinyl, pyrimidinyl, thiazolyl and pyridazinyl.

V

According to some embodiments of the present application, the structural unit of the compound of formula (IV) or the compound of formula (VI) is selected from the group consisting of

preferably / the structural unit is selected from the group consisting of ( ^R i)

more preferably the structural unit is selected from the group consisting of

and most preferably the structural unit is selected from the group consisting of

The present application also relates to a compound of formula (I-a) or a compound of formula (I-b), its pharmaceutically acceptable salt, stereoisomer or geometric isomer

where ring A, n, R1, R2, R ₃ , R4 and R5 are defined above.

In some embodiments, the structural unit is as defined above.

The present application also relates to a compound of formula (II-a) or a compound of formula (II-b), its pharmaceutically acceptable salt, stereoisomer or geometric isomer

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where ring A, n, R _b R ₂ and R ₅ are defined above.

In some embodiments, the structural unit

defined above.

The present application also relates to a compound of formula (III-a), formula (III-b), formula (IV-a) or formula (IV-b), a pharmaceutically acceptable salt, stereoisomer or geometric isomer thereof.

(IV-a) (IV-b) where n, R _b R ₂ , R5 and ring C are defined above.

ⁿ 7 s)

In some embodiments, the structural unit is as defined above.

The present application also relates to a compound of formula (V-a), formula (V-b), formula (VI-a) or formula (VI-b), a pharmaceutically acceptable salt, stereoisomer or geometric isomer thereof.

In some embodiments, the structural unit

defined above.

The present application also relates to a compound selected from the following structures, its pharmaceutically acceptable salt, tautomer, stereoisomer or geometric isomer

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application the compound of formula (I) is selected

According to some embodiments of the present, from the group consisting of

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- 23 040139

- 24 040139

- 25 040139

or a pharmaceutically acceptable salt thereof.

In another aspect, the present application also relates to a pharmaceutical composition containing a compound of formula (I), a pharmaceutically acceptable salt, tautomer, stereoisomer or geometric isomer thereof. In some embodiments, the pharmaceutical composition of the present application further comprises a pharmaceutically acceptable excipient, carrier, or diluent. In some embodiments, the pharmaceutical composition of the present application further comprises a pharmaceutically acceptable carrier, excipient, or adjuvant.

According to a further aspect, the present application also relates to a method of treating multiple myeloma in a mammal, comprising administering to a mammal, preferably a human, in need thereof, a therapeutically effective amount of a compound of formula (I), a pharmaceutically acceptable salt, tautomer, stereoisomer, or geometric isomer thereof, or a pharmaceutical composition thereof. In a further aspect, the present application relates to a method for the prevention or treatment of multiple myeloma, the method comprising administering to a subject in need thereof a compound of formula (I), a pharmaceutically acceptable salt, tautomer, stereoisomer, or geometric isomer thereof, or a pharmaceutical composition thereof.

According to another aspect, the present application also relates to the use of a compound of formula (I), a pharmaceutically acceptable salt, tautomer, stereoisomer or geometric isomer thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for the prevention or treatment of multiple myeloma.

In yet another aspect, the present application relates to a compound of formula (I), a pharmaceutically acceptable salt, tautomer, stereoisomer or geometric isomer thereof, or a pharmaceutical composition thereof, for use in the prevention or treatment of multiple myeloma. According to another aspect, the present application also relates to the use of a compound of the formula (I), its pharmaceutically acceptable salt, tautomer, stereoisomer or geometric isomer, or its pharmaceutical composition in the prevention or treatment of multiple myeloma.

According to another aspect, the present application also relates to a compound of formula (I), a pharmaceutically acceptable salt, tautomer, stereoisomer or geometric isomer thereof, or a pharmaceutical composition thereof, for use in the prevention or treatment of multiple myeloma.

In an additional aspect, the compound of formula (I) according to the present application can be obtained by a person skilled in the art in the following general schemes and using a standard method of the art:

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General scheme 1

1-6A

General scheme 3

1-ZA 1-4A 1-5A

BB-1A

General scheme 4

General scheme 5

3-1A 3-2A

General scheme 6

The compound according to the present application has a good antitumor effect against multiple myeloma and good oral bioavailability, and also has a therapeutic potential for resistance of multiple myeloma to bortezomib.

Definition

Unless otherwise noted, the following terms and expressions used in the present description have the following meanings. A particular term or expression should not be considered obscure or undefined unless specifically defined. They should be understood according to their general meaning. As used herein, the trade name refers to the respective product or its active ingredient.

The term pharmaceutically acceptable refers to a compound, substance, composition and/or dosage form that is applicable in contact with human and animal tissues without undue toxicity, irritation, allergic reaction or other problems or complications based on a thorough medical evaluation and in accordance with an acceptable balance of benefits. /risk.

The dotted line (—) in structural units or groups according to the present application refers to a covalent bond.

If the covalent bond in some structural units or groups according to this application

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O (for example, the dotted line (-) in ^^) is not associated with a specific atom, which means that a covalent bond can be associated with any atom in structural units or groups, since the theory of valence bonds is not violated.

T 5 O 5

Thus, for example, a structural unit includes

The term pharmaceutically acceptable salt refers to a salt of a compound of this application that is derived from a compound with the specific substituents disclosed herein and a relatively non-toxic acid or base. If the compound according to the present application contains a relative acidic functional group, a base addition salt can be obtained by bringing the compound into contact with a sufficient amount of base. The pharmaceutically acceptable base addition salt includes sodium, potassium, calcium, ammonium, organic ammonium or magnesium salt or the like. If the compound according to the present application contains a relatively basic functional group, an acid addition salt can be obtained by bringing the compound into contact with a sufficient amount of acid. Examples of the pharmaceutically acceptable acid addition salt include an inorganic acid salt, where the inorganic acid includes acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, hydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid. , phosphorous acid, and the like; and an organic acid salt, wherein the organic acid includes acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid , benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methylsulfonic acid, and the like; and also includes an amino acid salt (eg, arginine), and an organic acid salt such as glucuronic acid and the like. (See Berge et al., Pharmaceutical Salts, Journal of Pharmaceutical Science 66: 1-19 (1977)). Some specific compounds according to the present application contain alkaline and acidic functional groups to be able to turn into any base addition salts or acid addition salts.

Preferably, the parent compound is reduced by contacting the salt with a base or acid in the conventional manner and then separating the parent compound. The differences between the original form of the compound and its various salt forms lie in some physical properties. For example, the solubilities in a polar solvent are different.

As used herein, the pharmaceutically acceptable salt belongs to derivatives of the compound of the present application, wherein the parent compound is modified to form a salt with an acid or a base. Examples of a pharmaceutically acceptable salt include, without limitation, an inorganic or organic acid salt of a base (such as an amine), an alkali metal or an organic salt of an acid (such as a carboxylic acid), and the like. The pharmaceutically acceptable salt includes conventional non-toxic salts or quaternary ammonium salts of the parent compound, for example salts formed with a non-toxic inorganic or organic acid. Conventional non-toxic salts include, without limitation, those derived from inorganic acids and organic acids, where the inorganic acids or organic acids are selected from 2-acetoxybenzoic acid, 2-isethionic acid, acetic acid, ascorbic acid, benzenesulfonic acid, benzoic acid, bicarbonate, carbonic acid, citric acid, edetic acid, ethanedisulfonic acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, hydrobromic acid, hydrochloric acid, hydroiodate, hydroxynaphthoic acid, isethionic acid, lactic acid, dodecanesulfonic acid acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, nitric acid, oxalic acid, pamoic acid, pantothenic acid, phenylacetic acid, phosphoric acid, polygalacturonic acid, propionic acid, salicylic acid, c thearinic acid, a basic salt of acetic acid, succinic acid, aminosulfonic acid, sulfanilic acid, sulfuric acid, tannic acid, tartaric acid and p-toluenesulfonic acid.

The pharmaceutically acceptable salt of the present application can be synthesized from the starting compound containing an acid or base group by conventional chemical means. As a rule, the method of obtaining a salt includes: the interaction of such compounds in the forms of free acids or bases with a stoichiometric amount of the corresponding bases or acids in water or an organic solvent of a water-organic solvent mixture. In general, a non-aqueous medium such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile is preferred.

In addition to the salt form, there is a prodrug form for the present compound.

- 28 040139 invention. The prodrug of the compound described in the present invention was easily converted into the compound of the present invention by chemical changes under physiological conditions.

In addition, the prodrug can be converted to the compound of the present invention by chemical or biochemical means in an in vivo environment.

Some compounds according to the present application may exist in unsolvated or solvated forms, including hydrated forms. In general, the solvated form is similar to the unsolvated form, both of which are included within the scope of this application.

Some compounds according to the present application may contain asymmetric carbon atoms (stereocenter) or double bonds. Racemic isomers, diastereomers, geometric isomers and simple isomers are included within the scope of this application.

Unless otherwise noted, the absolute configuration of a stereocenter is represented by wedge and dashed links ''' λ one of the absolute configurations (e.g. one of and .·) of the stereocenter is represented by a wavy line ’ and the relative stereocenter configuration is represented by ''''

When a compound of the present application contains an olefinic double bond or other geometric center of asymmetry, E and Z geometric isomers are included, unless otherwise noted. Likewise, all tautomeric forms are included within the scope of this application.

The compound of the present application may exist in the form of a particular geometric or stereoisomeric isomer. This application contemplates all such compounds, including tautomers, cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, W)- isomers, as well as racemic mixtures and other mixtures, such as enantiomer- or diastereoisomer-enriched mixtures, are all included within the scope of this application. Other asymmetric carbons may exist in substituents such as alkyl. All such isomers and mixtures thereof are included within the scope of this application.

Optically active (R)- and (S)-isomers and (D)- and (L)-isomers can be obtained by chiral synthesis or chiral reagents or other conventional technologies. The enantiomer of the compound of the present application can be obtained by asymmetric synthesis or derivatizing action with chiral excipients, in which the resulting diastereomeric mixtures were isolated and the auxiliary groups were cleaved off to obtain the desired pure enantiomer. Alternatively, if the molecule contains an alkaline functional group (such as amino) or an acidic functional group (such as carboxyl), the molecule is reacted with an appropriate optically active acid or base to form a diastereomeric salt, the diastereomer is cleaved by well known conventional methods of the art, and then can be pure enantiomers were obtained. In addition, the separation of enantiomers and diastereomers was usually carried out by chromatography, which used a chiral stationary phase, and optionally combined with a chemical derivatization process (for example, carbamate is formed from an amine).

The compound of the present application may have an unusual proportion of atomic isotopes at one or more of the atoms that make up the compound. For example, the compound may be labeled with a radioactive isotope such as tritium ( ³ H), iodine-125 ( ¹²⁵ 1) or C-14 ( ¹⁴ C). All variants composed of isotopes of the compound disclosed in this application, whether radioactive or not, are included within the scope of this application.

The term pharmaceutical composition refers to a mixture of one or more compounds or their salts according to the present application and a pharmaceutically acceptable excipient. The purpose of the pharmaceutical composition is to facilitate the introduction of the compounds according to the present application into the body.

The term pharmaceutically acceptable carrier, pharmaceutically acceptable excipients or pharmaceutically acceptable adjuvant refers to those carriers, excipients or adjuvants that do not significantly affect the body and will not impair the biological activity and properties of the active compound. Suitable carriers, excipients or adjuvants are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and/or water-swellable polymers, hydrophilic or hydrophobic substances, gelatin, oils, solvents, water, and the like.

The term includes and variations thereof (such as includes or including) should be understood to have an open and non-exclusive meaning, i.e. includes without limitation.

The pharmaceutical composition of the present application can be obtained by combining the compounds of the present application with a suitable pharmaceutically acceptable adjuvant. For example, it may be formulated into solid, semi-solid, liquid or gaseous formulations such as tablets, pills, capsules, powders, granules, lozenges, ointments, syrups, emulsions, suspensions, solutions, suppositories, injections, volatiles, gels, microspheres. , aerosols, etc.

Exemplary routes of administration for a compound of the present application, or a pharmaceutically acceptable salt, tautomer, stereoisomer, or geometric isomer thereof, or a pharmaceutical composition thereof

- 29 040139 include, without limitation, oral, rectal, transmucosal, topical, transdermal, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration. Preferred routes of administration are oral administration and injection administration.

The pharmaceutical composition according to the present application can be prepared using a method known in the art, such as a conventional mixing method, a dissolution method, a granulation method, a dragee method, a grinding method, an emulsification method, a lyophilization method, and the like.

In some embodiments, the pharmaceutical composition according to the present application is in oral form. For oral administration, the pharmaceutical composition may be formulated by mixing the active compound with a pharmaceutically acceptable excipient or excipient well known in the art. Such excipient or adjuvant provides the formulation of the compound according to the present application into tablets, pills, lozenges, dragees, capsules, powders, granules, liquids, syrups, emulsions, gels, slurries, suspensions, etc., which are used for oral administration to the patient.

A solid pharmaceutical composition suitable for oral administration can be prepared by the conventional mixing, filling or tableting process. For example, oral compositions in solid form can be prepared by mixing the active compound with a solid excipient or excipient, optionally grinding the resulting mixture, optionally adding other appropriate excipients or excipients, and then processing the mixture into granules to form tablet cores. or dragee. Suitable adjuvants or adjuvants include, but are not limited to, fillers, binders, diluents, disintegrants, lubricants, glidants, sweeteners, flavors, and the like.

The pharmaceutical composition according to the present application may also be suitable for parenteral administration, such as a sterile solution, suspension, emulsion or lyophilized product in an appropriate unit dosage form. A suitable adjuvant such as a filler, buffer, or surfactant may be used.

The compound of formula (I) according to the present application can be administered daily at a dose of 0.01 mg/kg body weight to 200 mg/kg body weight in a single dose or in divided doses.

The term pharmaceutically acceptable carrier refers to any agent, carrier or carrier medium that is capable of delivering an effective amount of the active substance disclosed in this application, does not interfere with the biological activity of the active substance and does not cause toxic side effects on the recipient or patient. Typical carriers include water, oil and minerals, cream base, lotion base, ointment base, and the like. Such bases include suspensions, suspending agents, viscosity enhancers, transdermal penetration enhancers, and the like. Such agents are well known to those skilled in the art of cosmetics or topical medicines. Other carrier information can be found in Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are incorporated herein by reference.

The term adjuvant or excipient usually refers to a carrier, diluent and/or medium necessary to obtain an effective pharmaceutical composition.

With respect to a drug or pharmacologically active agent, the term "effective amount" or "therapeutically effective amount" means a sufficient amount of the drug or formulation that can achieve the desired effects without being toxic. In relation to an oral formulation in accordance with the present application, the term effective amount of one active substance in the composition means the amount necessary to achieve the desired effect, in combination with another active substance in the composition. The determination of an effective amount varies from person to person, depending on the age and general health of the subject, and also depends on the particular active substance. An appropriate effective amount in individual cases can be determined by a person skilled in the art in accordance with conventional tests.

The terms active ingredient, therapeutic agent, active substance or active agent refer to a chemical compound that can effectively treat the target disorder, disease or condition.

The term treatment process or treatment means that a compound or composition according to the present application is administered to prevent, alleviate or eliminate diseases or one or more symptoms associated with said diseases, and includes:

(i) preventing the manifestation of the disease or condition in mammals, especially if such

- 30 040139 mammals are susceptible to a disease or condition, but said disease or condition has not yet been diagnosed;

(ii) inhibition of the disease or condition, ie. inhibition of the development of the disease or condition;

(iii) alleviate the disease or condition, i. e. providing regression of the disease or condition.

The term "therapeutically effective amount" refers to an amount of a compound according to the present application that (i) treats or prevents a particular disease, condition, or disorder, (ii) relieves, alleviates, or eliminates one or more of the symptoms of a particular disease, condition, or disorder, or (iii) ) prevents or slows the onset of one or more symptoms of a particular disease, condition, or disorder described herein. The amount of a compound according to the present application, which determines a therapeutically effective amount, will vary depending on the nature of the compound, the condition and severity of the disease, condition or disorder, the route of administration, and the age of the mammal being treated. However, the amount may be determined according to normal practice by one of ordinary skill in the art based on his or her skill and the present disclosure.

The term optional or optional means that the event or condition described below may or may not occur and that the description includes cases in which the specified event or condition occurs and cases in which the specified event or condition does not occur.

The term substituted refers to one or more hydrogen atoms on a particular atom that are substituted by a substituent, including deuterium and hydrogen variants, and the valence state of a particular atom is normal and the compound obtained after substitution is stable. If the substituent is oxo (ie =O), which means that two hydrogen atoms are replaced, oxo substitution does not occur on the aromatic group. The term "optionally substituted" means that it may or may not be substituted, and unless otherwise noted, the type and number of substituents may be arbitrary, provided that this can be achieved in chemistry.

If any variable (eg, R or R ₁ ) occurs more than once in the composition or structure of the compound, the definition in each case is independent. Thus, for example, if a group is substituted with 0-2 R, the group may optionally be substituted with a majority of two R, and R in each case has an independent variant. As another example, each of the R1s in the structural unit

is independent and they may be the same or different. In addition, a combination of substituents and/or variants thereof is permitted if such a combination will result in a stable compound.

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

Unless otherwise noted, C _3-6 cycloalkyl-(CH ₂ ) _1-3 - includes C _3-6 cycloalkyl-CH ₂ -, C _3-6 cycloalkyl-(CH ₂ ) ₂ - and C _3-6 cycloalkyl -(CH ₂ ) ₃ -. Similarly, phenyl-(CH ₂ ) _1-3 - includes phenyl-CH ₂ -, phenyl-(CH ₂ ) ₂ - and phenyl-(CH ₂ ) ₃ -.

If one of the variables is a single link, it means that the two groups connected to it are directly related to each other. For example, if the L in A-L-Z is a single bond, this means that the actual structure is A-Z.

If the substituent is absent, this means that the substituent is not present. For example, if X is absent in A-X, this means that the structure is actually A. If a substituent can be connected to one or more atoms on the ring, the substituent can be connected to any atom on the ring. For example, a structural unit

means that the R substituent may be substituted by a hydrogen atom at any position on the cyclohexyl or cyclohexadiene ring. If the atom through which the given substituent is connected to the substitutable group is not designated, such a substituent may be bonded through any of its atom. For example, pyridyl as a substituent may be linked to the substituted group through any atom on the pyridine ring. If a given linking group does not show its linking direction, the linking direction is arbitrary. For example, a linking group L in can link ring A and ring B to form

represents -M-W-, then -M-Wva to the right, and with the formation

in the same direction as the left direction, opposite from left to right.

The combination of linking groups, substituents and/or variants thereof is only allowed if such a combination will result in a stable connection.

Unless otherwise noted, the term hetero is a heteroatom or a heteroatom group (i.e., a group containing a heteroatom) including atoms excluding carbon (C) and hydrogen (H), and

- 31 040139 groups containing such heteroatoms, for example, including oxygen (O), nitrogen (N), sulfur (S), silicon (Si), germanium (Ge), aluminum (Al), boron (B), -O- , -S-, =O, =S, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) ₂ and optionally substituted -C(=O)n(h)-, -N(H)-, -C(=NH)-, -S(=O) ₂ N(H)- or -S(=O) N(H)-.

Unless otherwise noted, the term ring refers to substituted or unsubstituted cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, aryl, or heteroaryl. Said ring includes a monocyclic, bicyclic, dicyclic, spiro, fused or bridged ring. The number of atoms in a ring is usually defined as the number of members that make up the ring. For example, a 5-7 membered ring refers to a ring formed by 5-7 atoms. Unless otherwise noted, the ring optionally contains 13 heteroatoms. Thus, a 5-7 membered ring includes, for example, phenyl, pyridinyl and piperidinyl. On the other hand, the term 5-7 membered heterocyclyl includes pyridyl and piperidinyl, but does not include phenyl. The term ring also includes a ring system containing at least one ring, where each ring independently corresponds to the above definition.

Unless otherwise noted, the term heterocycle or heterocyclyl refers to a stable monocyclic, bicyclic or tricyclic ring containing a heteroatom or heteroatomic group, they may be saturated, partially unsaturated or unsaturated (aromatic) and they contain carbon atoms and 1, 2, 3 or 4 a heteroatom which is independently selected from the group consisting of N, O and S, wherein any of the above heterocycle may be fused to a benzene ring to form a bicyclic ring. Nitrogen atoms and sulfur atoms may optionally be oxidized (ie NO and S(O) _p , p is 1 or 2). The nitrogen atoms may be substituted or unsubstituted (ie, N or NR, where R is H or other substituents as defined herein). The heterocycle can be attached to a pendant group of any heteroatoms or carbon atoms to form a stable structure. If the compound formed is stable, the heterocycle described herein may be substituted on its carbon or nitrogen atoms. The nitrogen atoms in the heterocycle are optionally quaternized. A preferred embodiment is one in which the total number of S and O atoms in the heterocycle is greater than 1, such heteroatoms are not adjacent to each other. Another preferred embodiment is one in which the total number of S and O atoms in the heterocycle is not more than 1. As used herein, the term aromatic heterocyclic group or heteroaryl refers to a stable 5-, 6-, 7-membered monocyclic or bicyclic or 7-, An 8-, 9-, or 10-membered bicyclic aromatic heterocyclyl that contains carbon atoms and 1, 2, 3, or 4 heteroatoms that are independently selected from the group consisting of N, O, and S. The nitrogen atoms may be substituted or unsubstituted (i.e. .N or NR if R is H or other substituents as defined herein). Nitrogen atoms and sulfur atoms may optionally be oxidized (ie NO and S(O) _p , p is 1 or 2). It should be noted that the total number of S and O atoms in an aromatic heterocycle is not more than 1. Bridged rings are also included in the definition of a heterocycle. When one or more atoms (ie, C, O, N, or S) are bonded to two non-adjacent carbon or nitrogen atoms, a bridged ring is formed. It should be noted that the bridge always converts a monocyclic ring into a tricyclic ring. In a bridged ring, a ring substituent may also be bridged.

Examples of heterocyclyl include, without limitation: acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzimidazolinyl, benzimidazolyl, benzomercaptophenyl, benzoxazolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzoisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-benzolyl, chromene, carboline, carbazolyl, decahydroquinolinyl, 2R,6R-1,5,2dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indoalkenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isoindolyl, isoindolinyl, isoquinolyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4- oxadiazolyl, oxazolidinyl, oxazolyl, hydroxyindolyl, pyrimidyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, benzoxanthinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidyl, piperidonyl, 4-pi peridonyl, piperonyl, pteridyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazolyl, pyridoimidazolyl, pyridothiazolyl, pyridyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolyl, 4H-quinolizinyl, quinolizinyl, quinolizinyl tetrahydrofuryl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thiantrenyl, thiazyl, isothiazolylthienyl, thienooxazolyl, thienothiazolyl, thienoimidazolyl, thienyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl and xanthenyl. Also included are fused ring and spiro compounds.

Unless otherwise noted, the term hydrocarbyl, or specific terms thereof (such as alkyl, alkenyl, alkynyl, aryl, etc.), by itself or as part of another substituent, is a straight, branched, or cyclic hydrocarbon group, or a combination thereof, which may

- 32 040139 be fully saturated (such as alkyl) or mono- or polyunsaturated (such as alkenyl, alkynyl and aryl), may be monosubstituted or polysubstituted, may be monovalent (eg methyl), divalent (eg methylene) or multivalent ( e.g. methine) may include divalent or multivalent atomic groups and contain a specified number of carbon atoms (e.g. C ₁ -C ₁₂ is 1 to 12 carbon atoms, C _1-12 is selected from C1, C ₂ , C ₃ , ^C 4, ^C 5, ^C 6, ^C 7, C8, ^C 9, ^C 10 ^{, C} 11 ^{and C} 12, ^and C 3-12 ^{is selected from C} 3, C 4, ^C 5, ^C 6, ^C 7, ^C 8, ^C 9, ^C 10, ^C 11 ^{and C} 12). The term ^hydrocarbyl includes, without limitation, aliphatic hydrocarbyl and aromatic hydrocarbyl. Aliphatic hydrocarbyl includes linear and cyclic aliphatic hydrocarbyl, and specifically includes, without limitation, alkyl, alkenyl, and alkynyl. The aromatic hydrocarbyl includes, without limitation, 6-12 membered aromatic hydrocarbyl such as phenyl, naphthyl, and the like. In some embodiments, the term hydrocarbyl is a straight or branched atomic group, or a combination thereof, which may be fully saturated, or mono- or polyunsaturated, and may include divalent and polyvalent groups. Examples of saturated hydrocarbon groups include, without limitation, homologues or isomers of methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl and n-amyl, n -hexyl, n-heptyl, n-octyl, and the like. The unsaturated hydrocarbyl contains one or more double bonds or triple bonds, and examples thereof include, without limitation, vinyl, 2-propenyl, butenyl, crotyl, 2-isopentenyl, 2-butadienyl, 2,4-pentadienyl, 3-(1,4 -pentadienyl), acetenyl, 1-propynyl and 3-propynyl, 3-butynyl, and the like. and higher homologues and isomers.

Unless otherwise noted, the term heterohydrocarbyl, or specific terms thereof (such as heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl, and the like), by itself or when combined with another term, is a stable straight, branched, or cyclic hydrocarbon group, or a combination thereof, which consists of a certain number of carbon atoms and at least one heteroatom. In some embodiments, the term heteroalkyl, by itself or when combined with another term, is a stable straight or branched hydrocarbon group, or combination thereof, that is composed of the specified number of carbon atoms and at least one heteroatom. In an exemplary embodiment, the heteroatom is selected from the group consisting of B, O, N, and S, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atoms are optionally quaternized. Heteroatoms or heteroatomic groups may be located at any internal positions of the heterohydrocarbyl, including the position where the hydrocarbyl is attached to the remainder of the molecule. However, the terms alkoxy, alkylamino, and alkylthio (or thioalkoxy) belong to common expressions and refer to those alkyl groups that are attached to the rest of the molecule via an oxygen atom, an amino group, or a sulfur atom, respectively. Examples include, without limitation -CH ₂ -CH ₂ -O-CH ₃ , -CH2-CH2-NHCH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2 -S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -CH2-CH=N-OCH ₃ and -CH=CH-N(CH ₃ ) _-CH3 . Mostly the two heteroatoms may be adjacent, such as -CH ₂ -NH-OCH ₃ .

Unless otherwise noted, the terms cyclohydrocarbyl, heterocyclohydrocarbyl, or specific terms thereof (such as aryl, heteroaryl, cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, and the like) alone or when combined with other terms, respectively, represent a cyclic hydrocarbyl or heterohydrocarbyl. In addition, with respect to heterohydrocarbyl or heterocyclohydrocarbyl (such as heteroalkyl and heterocycloalkyl), heteroatoms may occupy the position in which the heterocyclic ring is attached to the remainder of the molecule. Examples of cyclohydrocarbyl include, without limitation, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Non-limiting examples of heterocyclohydrocarbyl include 1-(1,2,5,6-tetrahydropyridinyl), 1-piperidyl, 2-piperidyl, 3-piperidyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuranilindol-3-yl, tetrahydrothiophen-2-yl, tetrahydrothiophen-3-yl, 1-piperazinyl and 2-piperazinyl.

Unless otherwise noted, the term alkyl refers to a straight or branched saturated hydrocarbyl which may be monosubstituted (eg -CH2F) or polysubstituted (eg -CF ₃ ) and may be monovalent (eg methyl), divalent (eg methylene) or polyvalent (for example, methine). Examples of alkyl include methyl (Me), ethyl (Et), propyl (e.g. n-propyl and isopropyl), butyl (e.g. n-butyl, isobutyl, sec-butyl and t-butyl), pentyl (e.g. n -pentyl, isopentyl and neopentyl) and the like. For example, the term C _1-3 alkyl refers to alkyl having 1 to 3 carbon atoms (such as methyl, ethyl, n-propyl, isopropyl). For example, the term C _1-6 alkyl refers to alkyl having 1 to 6 carbon atoms (such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc.).

Unless otherwise noted, cycloalkyl includes any stable monocyclic or polycyclic hydrocarbyl in which any carbon atom is saturated. Cycloalkyl may be monosubstituted or polysubstituted and may be monovalent, divalent, or polyvalent. Examples of cycloalkyl include, without limitation, cyclopropyl, norbornanyl,

[2.2.2]dicyclooctane, [4.4.0]dicyclodecane, etc.

Unless otherwise noted, cycloalkenyl includes any stable cyclic or polycyclic hydrocarbyl containing one or more unsaturated carbon-carbon double bonds at any position on the ring, which may be monosubstituted or polysubstituted, and may be monovalent, divalent or polyvalent. Examples of cycloalkenyl include, without limitation, cyclopentenyl, cyclohexenyl, and the like.

Unless otherwise noted, cycloalkynyl includes any stable cyclic or polycyclic hydrocarbyl containing one or more carbon-carbon triple bonds at any position on the ring, which may be monosubstituted or polysubstituted and may be monovalent, divalent or polyvalent.

Unless otherwise noted, the term halo or halogen per se or as part of another substituent refers to a fluorine (F), chlorine (Cl), bromine (Br), or iodine (I) atom. Moreover, the term haloalkyl is intended to include monohaloalkyl and polyhaloalkyl. For example, the term halo(C1- _C4 )alkyl is intended to include, without limitation, trifluoromethyl, 2,2,2-trifluoroethyl, 4chlorobutyl, 3-bromopropyl, and the like. Unless otherwise noted, examples of haloalkyl include, without limitation, trifluoromethyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl.

The term heteroalkyl is straight or branched alkyl which preferably has 1 to 14 carbon atoms, more preferably 1 to 10 carbon atoms, even more preferably 1 to 6 carbon atoms, and most preferably 1 to 3 carbon atoms in the chain, wherein one or more carbon atoms are replaced by a heteroatom selected from the group consisting of S, O and N. Exemplary heteroalkyl includes alkyl ethers, secondary alkylamines and tertiary alkylamines, amides, alkyl sulfides, and the like, including alkoxy, alkylthio , alkylamino. Unless otherwise noted, C1-6 heteroalkyl includes C1, _C2 , _C3 , _C4 , _C5 and _C6 heteroalkyl such as C1-6 alkoxy, C1-6 alkylthio and C1-6 alkylamino.

The term alkoxy refers to an alkyl group, as defined above, with the specified number of carbon atoms attached via an oxygen bridge. Unless otherwise noted, C1-6 alkoxy means C1, C ₂ , C ₃ , C ₄ , C ₅ and C ₆ alkoxy. Examples of alkoxy include, without limitation, methoxy, ethoxy, npropoxy, isopropoxy, n-butoxy, sec-butoxy, t-butoxy, n-pentyloxy, and S-pentoxy.

Unless otherwise noted, the term aryl is a polyunsaturated aromatic hydrocarbon substituent which may be monosubstituted or polysubstituted and may be monovalent, divalent or polyvalent. It may be monocyclic or polycyclic (eg 1-3 rings; at least one ring is aromatic). They are fused together or linked covalently. Aryl preferably contains 6 to 15 carbon atoms and more preferably 6 to 12 carbon atoms.

The term heteroaryl refers to an aryl containing 1 to 4 heteroatoms. In an exemplary embodiment, the heteroatom is selected from the group consisting of B, N, O and S, in which the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atoms are optionally quartetized. A heteroaryl may be linked to the remainder of the molecule via a heteroatom.

Non-limiting examples of aryl or heteroaryl include phenyl, naphthyl, biphenyl, pyrrolyl, pyrazolyl, imidazolyl, pyrazinyl, oxazolyl, phenyloxazolyl, isoxazolyl, thiazolyl, furanyl, thienyl, pyridyl, pyrimidinyl, benzothiazolyl, purinyl, benzimidazolyl, indolyl, isoquinolyl, quinoxalinyl, quinolyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl- 4-oxazolyl, 5oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3- pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzoimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. A substituent on any of the above aryl and heteroaryl ring systems is selected from the acceptable substituents described below.

Unless otherwise noted, when used in combination with other terms (eg, aryloxy, arylthio, arylalkyl), the term aryl includes aryl and heteroaryl rings as defined above. Thus, the term aralkyl is intended to include those aromatic groups (e.g., benzyl, phenethyl, pyridylmethyl, etc.) in which an aryl group is attached to an alkyl group, including an alkyl group in which a carbon atom (e.g., methylene ) is replaced by, for example, an oxygen atom, such as phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, and the like.

The term leaving group refers to a functional group or atom that can be replaced by another functional group or atom by a substitution reaction (such as a nucleophilic substitution reaction). By way of example, typical leaving groups include triflate; chlorine, bromine and iodine; a sulfonate group such as mesylate, tosylate, brosylate, p-toluenesulfonate and the like; and acyloxy such as acetoxy, trifluoroacetoxy and the like.

The term protecting group includes, without limitation, amino protecting group, carboxyl

- 34 040139 a protecting group, a hydroxyl protecting group and a mercapto protecting group. The term amino protecting group refers to a protecting group which is suitable for preventing the occurrence of side reactions at the nitrogen atom of an amino group. Exemplary amino protecting groups include, without limitation, formyl; acyl, for example an alkanoyl such as acetyl, trichloroacetyl or trifluoroacetyl; alkoxycarbonyl such as t-butoxycarbonyl (Boc); arylmethoxycarbonyl such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl such as benzyl (Bn), trityl (Tr) and 1,1di-(4'-methoxyphenyl)methyl; silyl such as trimethylsilyl (TMS) and t-butyldimethylsilyl (TBS); and so on. The term hydroxyl protecting groups refers to a protecting group which is suitable for preventing side reactions of the hydroxyl group. Exemplary hydroxy protecting groups include, without limitation, alkyl such as methyl, ethyl, and tert-butyl; acyl, eg alkanoyl, such as acetyl; arylmethyl such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (benzhydryl, DPM); silyl such as trimethylsilyl (TMS) and t-butyldimethylsilyl (TBS); and so on.

The compound of the present application can be prepared by many synthetic methods that are well known to those skilled in the art, including the following specific embodiments, embodiments obtained by combining the specific embodiments with other chemical synthetic methods, and equivalent alternative methods that are well known to those skilled in the art. technology. Preferred embodiments include, without limitation, examples of the present application.

The solvents used in this application are commercially available. In this application, the following abbreviations have been used: aq. is water; HATU is O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; EDC is N(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride; m-CPBA is 3-chloroperoxybenzoic acid; equiv. is equivalent or equal in number; CDI is carbonyldiimidazole; DCM is dichloromethane; PE is petroleum ether; DIAD is diisopropyl azodicarboxylate; DMF is N,N-dimethylformamide; DMSO is dimethyl sulfoxide; EtOAc is ethyl acetate; EtOH is ethanol; MeOH is methanol; CBz is benzyloxycarbonyl which is an amino protecting group; BOC is t-butoxycarbonyl which is an amino protecting group; HOAc/AcOH is acetic acid; NaCNBH ₃ is sodium cyanoborohydride; k.t. is room temperature; O/N represents all night; THF is tetrahydrofuran; Boc2O is di-tert-butyl dicarbonate; TFA is trifluoroacetic acid; FA is formic acid; ACN is acetonitrile; Hepes is 4-(2-hydroxyethyl)-1-piperazinylethanesulfonic acid; HBSS is Hank's Balanced Salt Solution; DIPEA/DIEA is diisopropylethylamine; SOCl ₂ is thionyl chloride; CS2 is carbon disulfide; TsOH is p-toluenesulfonic acid; NFSI is N-fluoro-N-(phenylsulfonyl)benzenesulfonamide; NCS is 1-chloropyrrolidine-2,5-dione; n-Bu ₄ NF is tetrabutylammonium fluoride; iPrOH is 2-propanol; m.p. represents the melting point; LDA is lithium diisopropylamide; IPA is isopropanol; DEA is diethylamine; DCE is dichloroethane; TMSCl is trimethylchlorosilane; TBTU is O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate; TEA is triethylamine.

Supplier catalog names were used for commercially available compounds.

Examples

The present application is described in detail below with the help of the following examples, but is not intended to be interpreted as limiting. The present application has been described in detail in the present description and its specific embodiments are also disclosed in the present description. A person skilled in the art will understand that various changes and modifications to the embodiments of the present application will be made without deviating from the spirit and scope of the present application.

Reference Example 1: Fragment BB-1

- 35 040139

Synthesis path:

Stage 1: Synthesis of compound BB-1-2

To a solution of compound BB-1-1 (50.00 g, 344.59 mmol, 45.05 ml, 1.00 eq.) in DCM (300.00 ml) was added cyclohexylamine (68.35 g, 689.18 mmol , 78.56 ml, 2.00 eq.) and calcium chloride (38.24 g, 344.59 mmol, 1.00 eq.) in an ice bath. The reaction mixture was heated, stirred at room temperature for 12 hours and then filtered. The filtrate was concentrated under reduced pressure to give a crude product, which was then recrystallized using methanol (50 ml) to give compound BB-1-2.

Stage 2: Synthesis of compound BB-1-3

Acetyl chloride (1.23 g , 24.96 mmol, 1.10 eq.) dropwise in an ice bath. The reaction mixture was stirred at 0° C. for 1 h, and then compound BB-1-2 (5.00 g, 22.69 mmol, 1.00 eq.) was added thereto. The resulting reaction mixture was warmed to room temperature, stirred continuously for 12 hours, and then concentrated under reduced pressure to obtain a crude product, which was then purified by silica gel column chromatography (mobile phase: dichloromethane:methanol = 50:1 - 20:1) to obtain compound BB-1-3.

Stage 3: Synthesis of compound BB-1-6

Compound BB-1-5 (30.00 g , 247.52 mmol, 1.02 eq.), anhydrous magnesium sulfate (145.58 g, 1.21 mol, 5.00 eq.) and p-toluenesulfonic acid (6.08 g, 24.19 mmol, 0. 10 equiv.) at room temperature. The reaction mixture was heated to 40° C., stirred continuously for 12 hours and then filtered. The filtrate was concentrated under reduced pressure to obtain a crude product, which was then purified by silica gel column chromatography (mobile phase: petroleum ether: ethyl acetate=100:1-20:1) to obtain compound BB-1-6.

Stage 4: Synthesis of compound BB-1-7

Bis(pinacolato)diborone (56.07 g, 220.79 mmol, 1.10 eq.), compound BB-1-3 (2.70 g, 10.04 mmol, 0.05 eq.) and sodium tert-butoxide (5.79 g, 60.22 mmol, 0.30 eq. .) at room temperature. The reaction mixture was stirred at room temperature for 96 hours under nitrogen gas protection and then concentrated under reduced pressure to give a crude product, which was then purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate=100:1-20: 1) to obtain compound BB-1-7.

Step 5: Synthesis of Compound BB-1

To a solution of compound BB-1-7 (46.00 g, 144.97 mmol, 1.00 eq.) in dioxane (120.00 ml) was added a solution of hydrochloric acid in dioxane (4 M, 80.00 ml, 2. 21 eq.) at room temperature. The reaction mixture was stirred at room temperature for 12 hours and then concentrated under reduced pressure to give a crude product. A solvent mixture of petroleum ether and ethyl acetate (petroleum ether:ethyl acetate = 5:1, 100 ml) was added to the crude product, stirred for 15 minutes, and then filtered. The filter cake was washed with methyl tert-butyl ether (100 ml) to give compound BB-1.

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 7.97 (s, 3H), 2.62-2.66 (m, 1H), 1.51-1.72 (m., 1H), 1.43-1.50 (m., 2H), 1.23 (s, 12H), 0.85 (d, J= 6.4 Hz, 6H).

- 36 040139

Example 1

Synthesis path:

Step 1: Synthesis of Compound 1-2

To a stirred solution of compound 1-1 (15.00 g, 74.55 mmol, 1.00 eq.) and methanol (150.00 ml) was added TMSCl (40.49 g, 372.75 mmol, 5.00 eq.) at 0°C, and then the reaction mixture was stirred at room temperature for 12 h under the protective action of nitrogen gas. The resulting reaction mixture was concentrated under reduced pressure to give the crude product, i.e. connections 1-2.

1H NMR: (400 MHz, METHANOL^) δ 5.18 (t, J=9.03 Hz, 1H), 4.15 (q, J=9.29 Hz, 1H), 3.88-4.00 (m, 1H), 3.79-3.86 (m, 3Н), 2.60-2.87 (m, 1H), 2.60-2.87 (m, 1H).

Step 2: Synthesis of Compound 1-3

To a solution of p-fluorophenylboronic acid (2.21 g, 15.83 mmol, 1.20 eq.) in acetonitrile (35.00 ml) was added compound 1-2 eq., hydrochloride), 4A molecular sieve (1.00 g), Cu(OAc) ₂ (2.64 g, 14.51 mmol, 1.10 eq.) and TEA (5.34 g, 52.76 mmol , 7.32 ml, 4.00 eq.) at room temperature. The reaction mixture was heated to 80° C. and stirred continuously for 12 hours. The reaction mixture was then filtered and the filtrate was concentrated. The obtained residue was purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate = 10:1) to obtain compound 1-3.

1H NMR: (400 MHz, CHLOROFORM-d) δ 6.87-6.97 (m, 2H), 6.44-6.52 (m, 2H), 4.45 (dd, J=7.65, 8.66 Hz, 1H), 4.00 (ddd, J= 3.76, 6.78, 8.53 Hz, 1H), 3.82 (s, 3H), 3.62-3.71 (m, 1H), 2.49-2.69 (m, 2H). MS (ESI) m/z: 210.0 [M+1].

Stage 3: Synthesis of compounds 1-4

LiOH -H ₂ O (401.11 mg, 9.56 mmol, 5.00 eq.) in an ice bath. The reaction mixture was stirred at room temperature for 1 hour and then adjusted to pH = 3 with 1 N dilute hydrochloric acid. The mixture solution was then concentrated and extracted with ethyl acetate. The organic phases were combined and concentrated to give the crude compound 1-4, which was used immediately in the next step. MS (ESI) m/z: 195.9 [M+1].

Step 4: Synthesis of Compound 1-5

To a solution of compound 1-4 (370.00 mg, 1.90 mmol, 1.00 eq.) in DMF (5.00 ml) was added methylphenylalaninate hydrochloride (491.75 mg, 2.28 mmol, 1.20 eq. ), TBTU (732.06 mg, 2.28 mmol, 1.20 eq.) and DIEA (982.22 mg, 7.60 mmol, 1.33 ml, 4.00 eq.) at -10°C. The reaction mixture was stirred at −10° C. to 0° C. for 1 hour, and then a saturated ammonium chloride aqueous solution (10 ml) was added thereto. The aqueous phase was extracted with ethyl acetate, and the organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and then concentrated. The resulting crude product was purified by silica gel column chromatography (mobile phase: petroleum ether: ethyl acetate = 1:1) to obtain compound 1-5.

1H NMR: (400 MHz, CHLOROFORM-d) δ 7.21-7.33 (m, 4H), 7.13-7.19 (m, 2H), 6.87-6.95 (m, 2H), 6.386.46 (m, 2H), 4.88- 4.96 (m, 1H), 4.20 (dd, J=7.78, 9.03 Hz, 1H), 3.82 (ddd, J=3.39, 6.96, 8.47 Hz, 1H), 3.71 (s, 3H), 3.60-3.67 (m, 1H), 3.29 (dd, J=5.65, 13.93 Hz, 1H), 3.07 (dd, J=7.78, 14.05 Hz, 1H), 2.45-2.56 (m, 1H), 2.03-2.17 (m, 1H) MS ( ESI) m/z: 357.1 [M+1].

Step 5: Synthesis of Compound 1-6

To a solution of compound 1-5 (500.00 mg, 1.40 mmol, 1.00 equiv.) in a mixture of THF (2.00 ml), MeOH (2.00 ml)

- 37 040139 and H2O (1.00 mL) LiOH-H2O (293.72 mg, 7.00 mmol, 5.00 eq.) was added at room temperature. The reaction mixture was stirred at room temperature for 12 hours and then adjusted to pH=3 with 1 N dilute hydrochloric acid. The resulting mixture solution was concentrated and then extracted with ethyl acetate. The organic phases were combined and concentrated to give the crude compound 1-6, which was used immediately in the next step. MS (ESI) m/z: 343.1 [M+1].

Step 6: Synthesis of Compound 1-7

To a solution of compound 1-6 (250.00 mg, 730.23 µmol, 1.00 eq.) in DMF (5.00 ml) eq., HCl), TBTU (304.80 mg, 949.29 µmol, 1.30 eq.) and DIEA (377.50 mg, 2.92 mmol, 510.13 µl, 4.00 eq.) at - 10°C. The reaction mixture was stirred at −10° C. to 0° C. for 1 hour, and then a saturated ammonium chloride aqueous solution (10 ml) was added thereto. The aqueous phase was extracted with ethyl acetate, and the organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and then concentrated to give the crude product of compound 1-7. MS (ESI) m/z: 538.3 [M+1].

Step 7: Synthesis of Compound WX-193

To a solution of compound 1-7 (390.00 mg, 725.62 µmol, 1.00 eq.) in methanol (5.00 ml) was added isobutylboronic acid (517.79 mg, 5.08 mmol, 7.00 eq. ) and an aqueous solution of HCl (1 M, 51.87 μl, 2.00 eq.) in an ice bath. The reaction mixture was warmed to room temperature and stirred continuously for 4 hours. The resulting reaction mixture was concentrated under reduced pressure to give the crude product, which was then separated and purified by preparative HPLC (0.225% FA) followed by SFC to give compound WX-193 .

1H NMR: (400 MHz, METHANOL-d ₄ ) δ 7.33 - 7.14 (m, 5H), 6.86 (br t, J=8.7 Hz, 2H), 6.63 - 6.45 (m, 2H), 4.77 (brt, J= 8.0 Hz, 1H), 4.66 - 4.58 (m, 1H), 4.52 - 4.41 (m, 1H), 3.17 - 2.97 (m, 4H), 2.65 (br t, J=7.5 Hz, 1H), 2.24 - 1.98 ( m, 2H), 1.34 (dt, J=6.7, 13.2 Hz, 1H), 1.13 (br t, J=7.4 Hz, 2H), 0.83 (br t, J=6.7 Hz, 6H)o MS (ESI) m /z: (M-17)438.2.

Separation method by SFC method:

Column: AD 250 mmx30 mm, 5 µm

Mobile phase: A: carbon dioxide; B: ethanol (containing 0.1% aqueous ammonia), elution gradient B%: 15% ~ 15%

Flow rate: 50 ml/min

Column temperature: 40°C

Compound WX-193 was the second peak in high performance chiral liquid column chromatography.

Compounds WX-268, WX-301, WX-351, WX-355, WX-365, WX-373, WX-381 and WX-385 were synthesized using the same method, and the separation conditions are as follows:

- 38 040139

No. connect- opinions Connection structure MS-17 'NMR Separation conditions WX-2 68 n 1 A n^n^ _n A _b , _0H ΛΑ 0 ⁿ OH F 362.1 Ή NMR (400 MHz, METHANOL-d4) δ 6.91 (br s, 2H), 6.49 (br s, 2H), 4.60 (br s, 1H), 4.00-4.40 (m, 1H), 3.50-3.97 (m, 2H), 2.95-3.19 (m, 3H), 2.69 (br s, 1H), 2.49 (br s, 1H), 2.03 (br s, 1H), 1.59 (br s, 3H), 0.93 (br s, 9H ) SFC separation, column: OJ (250 mm* 30 mm, 5 µm); mobile phase: A: carbon dioxide, B: ethanol, gradient elution V%: 15%-15%; speed flow: 60ml/min, its peak position was the second peak in high performance chiral liquid column chromatography.

- 39 040139

WX-3 01 η H 9 A n-^n^ _n A _b ,oh θ 0 ^H OH F 348.0 Ή NMR (400 MHz, METHANOL-d4) δ 6.76-6.93 (m, 2H), 6.63 (br d, J=4.52 Hz, 2H), 4.59 (br s, 5H), 4.08 (br d, J=10.29 Hz, 1H), 2.73 (br s, 1H) , 2.07-2.40 (m, 2H), 1.52-1.75 (m, 1H), 1.31 (br d, J=16.81 Hz, 2H), 0.80-0.97 (m, 6H) SFC separation, column: AD (250 mm x 30 mm, 10 µm); mobile phase: A: carbon dioxide, B: ethanol, gradient elution V%: 25%-25%; speed flow: 50 ml/min; its peak position was the second peak in high performance chiral liquid column chromatography. WX-3 51 h? A n^n^ _n A _b ,oh fA° ^{H 6h} F ₃ C 432.1 Ή NMR (400 MHz, METHANOL-d4) δ 7.38-7.57 (m, 2H), 6.69 (br d, J=8.53 Hz, 1H), 4.41-4.55 (m, 1H), 4.01-4.18 (m, 3H), 2.59-2.84 (m, 2H), 2.38-2.51 (m, 1H), 1.53-1.73 (m, 1H), 1.27-1.43 (m, 2H), 0.92 (d, J=6.78 Hz, 6H) Separation by preparative HPLC, column: Xtimate C18 150x25 mm, 5 µm; mobile phase: A: water (0.225% FA), B: methanol, gradient elution V%: 70%-100%, its retention time was 13 min using the high performance liquid column chromatography method. WX-3 55 η H 9 A A>-f ^{0 H} ° ^H F ₃ C 416.1 Ή NMR (400 MHz, METHANOL-d4) δ 7.16-7.42 (m, 2Η), 6.59-6.80 (m, 1H), 4.56-4.77 (m, 1H), Separation by preparative HPLC, column: Xtimate C18 150x25 mm, 5 µm; mobile phase: A:

- 40 040139

3.96-4.33 (m, 4H), 2.42-2.81 (m, 3H), 1.64 (br dd, J=6.53, 13.55 Hz, 1H), 1.20-1.48 (m, 2H), 0.92 (br d, >5.52Hz, 6H) water (0.225% FA), B: acetonitrile, elution gradient B%: 75%-85%, its retention time was 13.0 min in the high performance liquid column chromatography method. WX-3 65 ΓΛ ^H ίι f ⁴ n^n^ _n A _b , _oh ° ° ^H N ^Z 372.9 Ή NMR (400 MHz, METHANOL-d4) δ 7.28-7.44 (m, 2H), 6.64 (t, J=8.78 Hz, 1H), 4.73-4.80 (m, 2H), 3.96-4.32 (m, 4H), 2.62-2.81 (m, 2H), 2.38- 2.57 (m, 1H), 1.64 (qd, J=6.86, 13.55 Hz, 1H), 1.23-1.43 (m, 2H), 0.93 (d, J=6.53 Hz, 6H) SFC separation, column: AS (250 mm* 30 mm, 5 µm); mobile phase: A: carbon dioxide, B: ethanol, gradient elution V%: 15%-15%; its peak position was second peak in high performance chiral liquid column chromatography. WX-3 73 r\ H 9 AF N' ^R yN^< _N A _B ,OH A^f ^{0 H} ° ^H Cl 400.1 Ή NMR (400 MHz, METHANOL-d4) δ 6.81-7.05 (m, 2Η), 4.30-4.43 (m, 1H), 3.94-4.35 (m, 4H), 2.52-2.81 (m, 2H), 2.32-2.50 (m, 1H), 1.64 (qd, J=6.72, 13.46 Hz, 1H), 1.26-1.49 (m, 2H), 0.93 (dd, J=2.26, 6.53 Hz, 6H) Separation by preparative HPLC, column: Xtimate C18 150x25 mm, 5 µm; mobile phase: A: water (0.225% FA), B: methanol, gradient elution V%: 72%-82%, its retention time was 12.0 min with the method

- 41 040139

high performance liquid column chromatography. WX-3 81 π _n ? A n^n^ _n A _b , _0H Qhci ^{0 H 6h} F 381.9 Ή NMR (400 MHz, METHANOL-d4) δ 7.08 (dd, J=2.76, 8.28 Hz, 1H), 6.88-7.00 (m, 1H), 6.61 (dd, J=5.02, 9.03 Hz, 1H), 4.60 ( br t, J=8.28 Hz, 1H), 4.30-4.43 (m, 1H), 4.03-4.17 (m, 2H), 3.79 (q, J=7.86 Hz, 1H), 2.35-2.79 (m, 3H), 1.56-1.73 (m, 1H), 1.25-1.40 (m, 2H), 0.91 ( d, J=6.53Hz, 7H) Separation by preparative HPLC, Column: Xtimate C18 150x25 mm, 5 µm; mobile phase: A: water (0.225% FA), B: methanol, gradient elution V%: 65%-85%, its retention time was 13.0 min in the high performance liquid column chromatography method. WX-3 85 η H 9 AfyCF? ^HάΗCl 432.1 Ή NMR (400 MHz, METHANOL-d4) δ 7.29-7.59 (m, 2H), 6.62 (d, J=9.03 Hz, 1H), 4.54-4.78 (m, 1H), 4.26 (br s, 1H), 3.84-4.15 (m, 3H), 2.52-2.82 (m, 2H), 2.32-2.49 (m, 1H), 1.63 (td, J=6.71, 13.18 Hz, 1H), 1.23-1.42 (m, 2H), 0.91 (br d, J=6.27 Hz, 6H) Separation by preparative HPLC, column: Xtimate C18 150x25 mm, 5 µm; mobile phase: A: water (0.225% FA), B: methanol, gradient elution V%: 70%-100%, its retention time was 9.5 minutes using the high performance liquid column chromatography method.

Example 2

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Synthesis path:

Step 1: Synthesis of Compound 2-2

To a solution of compound 1-2 (1.00 g, 6.60 mmol, 1.00 eq.) and 2-chloro-4-phenylpyrimidine (1.26 g, 6.60 mmol, 1.00 eq.) in EtOH (20.00 ml) was added DIPEA (2.56 g, 19.80 mmol, 3.46 ml, 3.00 eq.) and Na ₂ CO ₃ (2.10 g, 19.80 mmol, 3.00 eq. .) at room temperature. The reaction mixture was heated to 40° C. and stirred for 12 hours. The solvent was then removed by rotary evaporation under reduced pressure, the residue was diluted with water (20 ml) and then extracted with ethyl acetate. The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered and then concentrated. The resulting residue was separated by preparative chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to obtain compound 2-2. MS (ESI) m/z: 269.9 [M+1].

Step 2: Synthesis of Compound 2-3

LiOH-H ₂ O ₍ 23 .37 mg, 0.57 mmol, 1.00 eq.) at room temperature. The reaction mixture was stirred at room temperature for 4 hours and then adjusted to pH=6-7 with 1N dilute hydrochloric acid. The resulting mixture solution was concentrated to give the crude product of compound 2-3, which was used immediately in the next step. MS (ESI) m/z: 255.9 [M+1].

Step 3: Synthesis of Compound 2-4

To a solution of compound 2-3 (150.00 mg, 0.59 mmol, 1.00 eq.) in DMF (2.00 ml) was added methyl phenyalaninate hydrochloride (126.37 mg, 0.71 mmol, 1.20 eq. ), TBTU (377.34 mg, 1.18 mmol, 2.00 eq.) and DIPEA (303.77 mg, 2.35 mmol, 0.41 ml, 4.00 eq.) at -20°C. The reaction mixture was stirred at -20°C to 0°C for 2 h, and then water (10 ml) was added thereto. The aqueous phase was extracted with ethyl acetate and the organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered and then concentrated to give the crude product, which was then separated and purified by preparative chromatography (mobile phase: petroleum ether: ethyl acetate = 2:1 ) to give compound 2-4.

Step 4: Synthesis of Compound 2-5

LiOH-H ₂ O (75.56 mg, 1.80 mmol, 3.00 eq.) at room temperature. The reaction mixture was stirred at room temperature for 12 hours and then adjusted to approximately pH=3 with 1 N dilute hydrochloric acid. The resulting mixture solution was concentrated and extracted with ethyl acetate. The organic phases were combined and concentrated to give the crude compound 2-5, which was used immediately in the next step. MS (ESI) m/z: 403.5 [M+1].

Step 5: Synthesis of Compound 2-6

To a solution of compound 2-5 (260.00 mg, 646.06 μmol, 1.00 eq.) in DMF (5.00 ml) was added compound BB-1 (193.49 mg, 775.27 μmol, 1.20 eq.), TBTU (311.15 mg, 969.09 µmol, 1.50 eq.) and DIPEA (333.99 mg, 2.58 mmol, 451.33 µl, 4.00 eq.) at -10° WITH. The reaction mixture was stirred at −10° C. to 0° C. for 1 hour, and then a saturated ammonium chloride aqueous solution (10 ml) was added thereto. The aqueous phase was extracted with ethyl acetate, and the organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and then concentrated to give the crude product of compound 2-6. MS (ESI) m/z: 599.2 [M+1].

Step 6: Synthesis of Compound WX-174

To a solution of compound 2-6 (380.00 mg, 634.88 µmol, 1.00 eq.) in MeOH (3.00 ml) was added isobutylboronic acid (478.92 mg, 4.70 mmol, 7.40 eq. ) and an aqueous solution of HCl (1 M, 1.27 ml, 2.00 eq.) at 0°C. The reaction mixture was stirred at 0 to 20° C. for 1 hour and then concentrated under reduced pressure to give a crude product which was then separated by preparative HPLC (0.225% FA) to give compound WX-174.

Ή NMR (400 MHz, METHANOL-d ₄ ) δ 8.38 (br d, J=5.3 Hz, 1H), 8.21-8.03 (m, 2H), 7.59-7.45 (m, 3H),

- 43 040139

7.36-7.05 (m, 6H), 4.20-3.95 (m, 2H), 3.23-2.93 (m, 2H), 2.67-2.44 (m, 2H), 2.23 (br s, 1H), 1.49-1.07 (m, 3H),

0.83 (br t, J=6.0 Hz, 6H). MS (ESI) m/z: 497.9 [M-17].

Method of SFC Analysis of Compound WX-174:

Column: AS 150 mm x 4.6 mm, 5 µm

Mobile phase: A: carbon dioxide; B: ethanol (containing 0.05% diethanolamine), elution gradient B%: 5% ~ 40%

Flow rate: 3 ml/min

Column temperature: 40°C

The retention time of WX-174 was 2.592 minutes on high performance chiral liquid column chromatography.

Connections WX-260, WX-306, WX-308, WX-311, WX-313, WX-317, WX-319, WX-327, WX-329, WX-367, WX-379, WX-387 and WX -393 was synthesized using the same method and the conditions for their separation are as follows: _________________________________________________________________

No. connect- opinions Connection structure MS-17 'NMR Separation conditions WX-2 60 Hmm ZI O-oo) - ¹ b / 404.2 Ή NMR (400 MHz, METHANOL-d4) δ 8.17 (s, 2H), 4.52-4.66 (m, 2H), 3.88-3.95 (m, 2H), 3.15 (td, J=E54, 3.20 Hz, 1H), 2.24-2.60 (m, 3H), 1.61-1.76 (m, 1H), 1.43-1.55 (m, 2H), 1.12-1.23 (m, 2H), 0.76 (d, J=6.53 Hz, 6H), 0.56- 0.67 (m, 1H), 0.24-0.41 (m, 2H), -0.08-0.06 (m, 2H). Preparative HPLC separation, column: Xtimate C18 150 mm *25 mm, 5 µm; mobile phase: A: water (0.225% FA), B: methanol, elution gradient B%: 60%-80%, its retention time was 7.0 min. high performance liquid column chromatography.

-44040139

WX-3 06 O-co) - ^T b / I 452.9 Ή NMR (400 MHz, METHANOL-d ₄ ) δ 8.35 (s, 1H), 7.98-8.12 (m, 1H), 6.86 (d, 7=9.29 Hz, 1H), 5.11-5.24 (m, 1H), 4.61 -4.73 (m, 1H), 4.18-4.41 (m, 2H), 2.83-2.97 (m, 1H), 2.75 (br dd, 7=6.27, 8.78 Hz, 1H), 2.48-2.62 (m, 1H), 1.81-1.92 (m, 1H), 1.58-1.72 (m, 2H), 1.26-1.42 (m, 2H), 0.89-0.96 (m, 6H), 0.74-0.87 (m, 1H), 0.43-0.59 (m , 2H), 0.13-0.24 (m, 2H) Preparative HPLC separation, column: Phenomenex Synergi C18 (150mm*30mm, 4um); mobile phase: A: water (0.225% FA), B: methanol), elution gradient B%: 70%-85%, 10 min); flow rate: 25 ml/min, its retention time was 2.8 min with the method high performance liquid column chromatography. WX-3 08 Ο H 9 A θ4 ok \7 CF ₃ 459.1 Ή NMR (400 MHz, METHANOL-d ₄ ) δ 7.38 (s, 1H), 4.84 (br s, 1H), 4.69 (t, 7=7.28 Hz, 1H), 4.08-4.19 (m, 1H), 4.03 ( q, 7=7.61 Hz, 1H), 2.67-2.79 (m, 2H), 2.51-2.65 (m, 1H), 1.78-1.90 (m, 1H), 1.58-1.72 (m, 2H), 1.31-1.38 ( m, 2H), 0.90-0.94 (m, 6H), 0.73-0.85 (m, 1H), 0.40-0.57 (m, 2H), 0.11-0.20 (m, 2H) Preparative HPLC separation, column: Phenomenex Synergi C18 (150 mmx30 mm, 4 µm); mobile phase: A: water (0.225% FA), B: methanol, elution gradient B%: 70%-85%, 10 min); flow rate: 25 ml/min, its retention time was 2.8 min

- 45 040139

with the method high performance liquid column chromatography. WX-3 And Xa o ZI _χ A O-oo ) - ^x ъ / 350.2 Ή NMR (400 MHz, METHANOL-d ₄ ) δ 8.35 (s, 2H), 4.76 (dd, 7=6.78, 9.29 Hz, 1H), 4.03-4.17 (m, 4H), 2.74 (br t, 7=7.65 Hz, 1H), 2.57-2.69 (m, 1H), 2.46 (qd, 7=7.72, 10.98 Hz, 1H), 1.59-1.71 (m, 1H), 1.31-1.41 (m, 2H), 0.92 (d, 7=6.53Hz, 6H) Preparative HPLC separation, column: Phenomenex Synergi С18 150 mm*30 mm, 4 microns; mobile phase: A: water (0.225% FA), B: methanol, elution gradient B%: 50%-75%, its retention time was 7.5 minutes with the method high performance liquid column chromatography. WX-3 13 GD n fl A /X o \u003d ⁿ OH yx F 364.1 Ή NMR (400 MHz, METHANOL-d4) δ 8.36 (s, 2H), 4.61-4.84 (m, 2H), 4.05-4.16 (m, 2H), 2.38-2.80 (br, 3H), 1.61-1.73 (m , 1Н), 1.48 (d, J=7.28 Hz, 3H), 1.29-1.40 (m, 2H), 0.95 (d, J=6.53 Hz, 7H). SFC separation, column: AD (250 mm*30 mm, 5 µm); mobile phase: A: carbon dioxide, B: ethanol, elution gradient B%: 15%-15%; his the peak position was the second peak in

- 46 040139

high performance chiral liquid column chromatography. WX-3 17 He ? A ν^ν^ _ν ^^οη A ° ^H ° ^H F 349.1 6.57 (dd, J=3.51, 9.03 Hz, 1H), 4.64 (br t, J=8.28 Hz, 1H), 4.05-4.25 (m, 2H), 3.90 (q, J=7.86 Hz, 1H), 2.74 (br s, 1H), 2.42-2.67 (m, 2H), 1.57- 1.75 (m, 1H), 1.36 (br t, J=7.03 Hz, 2H), 1.15 (d, J=6.27 Hz, 1H), 0.93 (br d, J=6.53 Hz, 6H) Separation by SFC method, column: Chiralpak AS-H 250mm*30mm, 5µm; mobile phase: A: carbon dioxide, B: ethanol (containing 0.1% NH3.H2O), elution gradient B%: 20%-20%; his the peak position was the second peak in high performance chiral liquid column chromatography. WX-3 19 h? ^A0HOH 294.1 Ή NMR (400 MHz, METHANOL-d4) δ 4.03-4.25 (m, 2Η), 3.87-3.98 (m, 1H), 3.42-3.52 (m, 1H), 2.76 (br s, 1H), 2.33-2.46 (m, 1H), 2.05-2.22 (m, 2H), 1.58-1.78 (m, 1H), 1.30-1.46 (m, 3H), 1.16-1.24 (m, 1H), 0.88-0.99 (m, 6H), Preparative HPLC separation, column: DuraShell 150mm*25mm, 5µm; mobile phase: A: water (0.225% FA), B: acetonitrile, elution gradient B%: 0%-30%, its time

- 47 040139

0.39-0.65 (m, 4H). retention was 6.5 min with the method high performance liquid column chromatography. WX-3 27 h? A 0 ^H OH %/N CF ₃ 405.1 'H NMR (400 MHz, METHANOL-d4) δ 7.43 (s, 1H), 4.84 (br s, 1H), 4.00-4.27 (m, 4H), 2.47-2.91 (m, 3H), 1.69 (td, J =6.62, 13.11 Hz, 1H), 1.28-1.47 (m, 2H), 0.94 (br d, J=6.27 Hz, 6H). Preparative HPLC separation, column: Phenomenex Synergi С18 150 mm*30 mm, 4 microns; mobile phase: A: water (0.225% FA), B: methanol, elution gradient B%: 50%-75%, its retention time was 9.4 min in the method high performance liquid column chromatography. WX-3 29 η h fl A / ⁼ ^ 0 ^H OH yy F ₃ C 399.1 Ή NMR (400 MHz, METHANOL-d4) δ 8.36 (br s, 1H), 7.83 (br d, J=8.78 Hz, 1H), 6.63 (br d, J=8.78 Hz, 1H), 4.77-4.84 (m , 1H), 4.01-4.26 (m, 4H), 2.49-2.84 (m, 3H), 1.68 (br dd, J=6.40, 12.92 Hz, 1H), 1.24-1.47 (m, Preparative HPLC separation, column: Phenomenex Synergi C18 150 mmx30 mm, 4 µm; mobile phase: A: water (0.225% FA), B: methanol, elution gradient B%:

- 48 040139

2H), 0.94 (br d, J=6.27 Hz, 6H) 55%-80%, its retention time was 9.2 minutes with the method high performance liquid column chromatography. WX-3 67 П h 9 A ν^ν^ _ν Α _β ,οη ^{0 H 0H} F ₃ C ^Z 400.1 Ή NMR (400 MHz, METHANOL-d4) δ 7.77 (dd, J=2.38, 9.41 Hz, 1H), 7.03 (d, J=9.29 Hz, 1H), 4.95-5.06 (m, 1H), 4.07-4.29 ( m, 4H), 2.53-2.86 (m, 3H), 1.61-1.80 (m, 1H), 1.27-1.49 (m, 2H), 0.86-1.00(m, 6H) Preparative HPLC separation, column: Xtimate C18 150 mm*25 mm, 5 µm; mobile phase: A: water (0.225% FA), B: methanol, elution gradient B%: 55%-85%, its retention time was 13.0 min. high performance liquid column chromatography. WX-3 79 ΓΛ H 9 AA^ _f ^{0 H} OH F 367.1 Ή NMR (400 MHz, METHANOL-d4) δ 7.89 (br s, 1H), 7.43 (br t, 7=9.54 Hz, 1H), 4.78-4.83 (m, 1H), 3.96-4.24 (m, 4H), 2.74 (br s, 1H), 2.63 (br d, 7=7.03 Hz, 1H), 2.46-2.58 (m, 1H), 1.65 (br d, 7=6.02 Hz, 1H), Preparative HPLC separation, column: Xtimate C18 (150 mmx25 mm, 5 µm); mobile phase: A: carbon dioxide, B: ethanol, gradient elution B%: 55%-85%, 9.5 min);

- 49 040139

1.35 (br t, 7=6.90 Hz, 2H), 0.92 (br d, 7=5.77 Hz, 6H) flow rate: 25 ml/min, its retention time was 4.0 min with the method high performance liquid column chromatography. WX-3 87 GL H 9 A (/A_ _f ^{0 H} OH N=/ 350.1 'H NMR (400 MHz, METHANOL-d4) δ 8.28 (br s, 1H), 7.98-8.16 (m, 2H), 4.93-5.10 (m, 1H), 4.32 (br d, J=5.52 Hz, 2H) , 4.01-4.23 (m, 2H), 2.70-2.84 (m, 1H), 2.37-2.67 (m, 2H), 1.53-1.72 (m, 1H), 1.20-1.42 (m, 2H), 0.88 (d, J=6.53Hz, 6H) Preparative HPLC separation, column: Xtimate C18 150 mm*25 mm, 5 µm; mobile phase: A: water (0.225% FA), B: methanol, elution gradient B%: 21%-51%, its retention time was 13.0 min. high performance liquid column chromatography. WX-3 93 η H 9 A \ N'V^^OH A^ _f ^{0 H} OH N ⁷ 391.1 ^X H NMR (400 MHz, METHANOL-d4) δ 8.08 (br s, 1H), 7.17-7.38 (m, 2H), 5.06 (dt, J=3.39, 6.21 Hz, 1H), 4.23-4.53 (m, 2H ), 4.11 (s, 2H), 2.30-2.84 (m, 3H), 1.56-1.75 (m, 1H), 1.23-1.45 (m, 2H), Preparative HPLC separation, column: Xtimate C18 150 mm*25 mm, 5 µm; mobile phase: A: water (0.225% FA), B: methanol, elution gradient B%:

- 50 040139

0.80-1.03 (m, 6H) 64%-74%, its retention time was 12.0 min with the method high performance liquid column chromatography.

3-6

WX-333

Step 1: Synthesis of Compound 3-3

To a solution of compound 3-1 (10 g, 77.46 mmol, 1.00 eq.) and compound 3-2 (20.13 g, 77.46 mmol, 1.00 eq.) in acetonitrile (200 ml) was added Ν,N-diisopropylethylamine (22.02 g, 170.40 mmol, 2.20 eq.) at room temperature. The reaction mixture was stirred at 100°C for 16 h, then cooled to room temperature and then added to ethyl acetate. The organic layer was further washed with water and brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated and then purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate=10:1) to obtain compound 3-3. MS (ESI) m/z: 227.9 [M+1].

Step 2: Synthesis of Compound 3-4

LiOHH ₂ O (6.65 g, 158.45 mmol, 5.00 eq.) at 0°C. The reaction mixture was stirred at room temperature for 1 hour and then concentrated under reduced pressure, diluted with water and ethyl acetate, and separated into layers. The aqueous layer was adjusted to pH=6 with 1N hydrochloric acid and then extracted with ethyl acetate. The organic phase was combined and washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give compound 3-4, which was used immediately in the next step. MS (ESI) m/z: 213.9 [M+1].

Step 3: Synthesis of Compound 3-5

To a solution of compound 3-4 (1.5 g, 7.04 mmol, 1.00 eq.) in dichloromethane (50 ml) was added glycine methyl ester hydrochloride (1.06 g, 8.44 mmol, 1.20 eq. , hydrochloride), TBTU (2.71 g, 8.44 mmol, 1.20 eq.) and Ν,N-diisopropylethylamine (3.64 g, 28.15 mmol, 4.90 ml, 4.00 eq.) at -10°C. The reaction mixture was stirred at -10°C to 0°C for 3 h, and then diluted with water (40 ml) and was extracted with dichloromethane. The organic phase was combined and washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and then purified by silica gel column chromatography (mobile phase: petroleum

-51 040139 ether:ethyl acetate=5:1) to give compound 3-5. MS (ESI) m/z: 284.9 [M+1].

Step 4: Synthesis of Compound 3-6:

LiOH-H ₂ O (369 03 mg, 8.79 mmol, 5.00 eq.) at 0°C. The reaction mixture was stirred at 0°C to 20°C for 2 h and then concentrated, diluted with water (3 ml) and separated into layers. The aqueous layer was adjusted to pH=6 with 1N hydrochloric acid and then extracted with ethyl acetate. The organic phase was combined and washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give compound 3-6, which was used immediately in the next step. MS (ESI) m/z: 270.9 [M+1]

Step 5: Synthesis of Compound 3-8

To a solution of compound 3-6 (0.26 g, 962.14 µmol, 1.00 eq.), compound 3-7 (437.84 mg, 1.15 mmol, 1.20 eq.) and TBTU (370, 71 mg, 1.15 mmol, 1.20 eq.) in dichloromethane (10 ml) was added Ν,N-diisopropylethylamine (273.56 mg, 2.12 mmol, 2.20 eq.) at -10°C. The reaction mixture was slowly warmed to room temperature and stirred continuously for 2 hours. The reaction mixture was then diluted with water (10 ml) and extracted with dichloromethane. The organic phase was combined and washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and then purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate = 1:1) to obtain compound 3-8. MS (ESI) m/z: 518.2 [M+1]

Step 6: Synthesis of Compound WX-333

To a solution of compound 3-8 (0.17 g, 328.56 µmol, 1.00 equiv.) in a mixture of methanol (4 ml) and n-hexane (6 ml) mmol, 7.00 eq.) and 1 M HCl (1.31 ml, 4.00 eq.) at 0°C. The reaction mixture was slowly warmed to room temperature, stirred continuously for 12 hours, and then concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC and then separated by SFC to give compound WX-333.

¹ H NMR (400 MHz, METHANOL-d ₄ ) δ 6.83 (br s, 2H), 6.61 (br s, 1H), 4.49 (br s, 1H), 4.10 (br s, 3H), 3.84 (br s, 1H), 2.75 (br s, 1H), 2.59 (br s, 1H), 2.48 (br s, 1H), 1.62 (br s, 1H), 1.30 (br s, 2H), 0.92 (br s, 6H) _OMS (ESI) m/z: 366.1 [M-17].

Method for Preparative HPLC Separation of Compound WX-333:

Column: Xtimate C18 150x25 mm, 5 µm;

Mobile phase: water (0.225%FA)-MeOH

Retention time: 9.5 min

Method for Preparative SFC Separation of Compound WX-333:

Column: C2 250 mmx30 mm, 10 µm

Mobile phase: A: carbon dioxide; B: methanol, elution gradient B%: 30%-30%

Flow rate: 60 ml/min

The peak position of the WX-333 compound was the second peak in high performance chiral liquid column chromatography.

Compound WX-391 was synthesized using the same method and its separation conditions are as follows:

- 52 040139

No. connect- nia Structure connections MS-17 'NMR Separation conditions WX-391 P n 9 A Ul ^{o n} on FF 384.1 Ή NMR (400 MHz, METHANOL-d ₄ ) δ 7.06 (dt, 7=7.28, 11.17 Hz, 1H), 6.53 (td, 7=7.94, 11.73 Hz, 1H), 4.60 (br s, 2H), 4.54 ( br t, 7=8.16 Hz, 1H), 4.03-4.17 (m, 3H), 3.87 (q, 7=7.53 Hz, 1H), 2.67-2.81 (m, 1H), 2.36-2.67 (m, 2H), 1.55-1.75 (m, 1H), 1.26-1.43 (m, 3H), 0.93 (dd, 7=1.63, 6.65 Hz, 6H) Separation by preparative HPLC, column: Xtimate C18 150x25 mm, 5 µm; mobile phase: A: water (0.225% FA), B: methanol, gradient elution: In%: 52%-82%, its retention time was 7.0 min using the high performance liquid column chromatography method.

Example 4

Synthesis path:

Step 1: Synthesis of Compound 4-3:

To a solution of compound 4-2 (4.37 g, 24.68 mmol, 2.84 ml, 1.00 eq) in DMSO (40 ml) was added compound 1-2 (5 g, 32.98 mmol, 1. 34 eq.) and DIEA (9.57 g, 74.03 mmol, 3.00 eq.). The reaction mixture was heated to 110° C. and stirred continuously for 3 hours, and then water (20 ml) was added thereto. The aqueous phase was extracted with ethyl acetate. The organic phase was combined and washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The resulting residue was purified by silica gel column chromatography (mobile phase: petroleum ether: ethyl acetate=1:0-5:1) to obtain compound 4-3. MS (ESI) m/z: 272.9 [M+1].

Step 2: Synthesis of Compound 4-4

To a solution of compound 4-3 (5.2 g, 19.10 mmol, 1.00 eq.) in THF (50 ml) was added 10% Pd/C (1 g, 9.55 mmol, 0.5 eq.) at room temperature. The reaction mixture was reacted under H ₂ (15 psi) for 12 hours and then filtered. The filtrate was concentrated, and the residue was purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate=1:0 1:1) to obtain compound 4-4. MS (ESI) m/z: 242.9 [M+1].

Step 3: Synthesis of Compound 4-5

To a solution of compound 4-4 (3.2 g, 13.21 mmol, 1.00 eq.) in ethanol (30.00 ml) was added fluoroboronic acid (7.73 g, 42.28 mmol, content 48%, 3 ,20 eq.) and isoamyl nitrite (1.70 g, 14.53 mmol, 1.10 eq.) at 0°C. The reaction mixture was stirred at 0° C. for 1.5 h, and then water (20 ml) was added thereto. The resulting mixture was extracted with ethyl acetate. The organic phase was combined and washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The resulting residue was purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate=1:0-3:1) to obtain compound 4-5. MS (ESI) m/z: 228.3 [M+1].

Step 4: Synthesis of Compound 4-6

To a solution of compound 4-5 (1.1 g, 4.84 mmol, 1.00 eq.) in a mixture of THF (4.00 ml), MeOH (4.00 ml) and H ₂ O (2.00 ml) added LiOH-H ₂ O (1.02 g, 24.21 mmol, 5.00 eq.) at 0°C. The reaction mixture was slowly warmed to room temperature, stirred continuously for 1 hour, and then adjusted to approximately pH=5 with 1N dilute hydrochloric acid. The resulting mixture solution was concentrated and extracted with ethyl acetate. The organic phase was combined and concentrated to give the crude product of compound 4-6, which was used immediately in the next step. MS (ESI) m/z: 214.0 [M+1].

Step 5: Synthesis of Compound 4-7

To a solution of compound 4-6 (0.68 g, 3.19 mmol, 1.00 eq.) in DCM (10.00 mL) was added glycine methyl ester hydrochloride (480.59 mg, 3.83 mmol, 1.20 eq.), TBTU (1.23 g, 3.83 mmol, 1.20 eq.) and DIEA (1.65 g, 12.76 mmol, 4.00 eq.) at -10°C. The reaction mixture was stirred at -10°C to 0°C for 0.5 h, and then 15 ml of water was added thereto. The resulting mixture was extracted with dichloromethane. The organic phase was combined and washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The resulting crude product was purified by silica gel column chromatography (mobile phase: petroleum ether: ethyl acetate=1:0-3:1) to obtain compound 4-7. MS (ESI) m/z: 284.9 [M+1].

Step 6: Synthesis of Compound 4-8

To a solution of compound 4-7 (0.5 g, 1.76 mmol, 1.00 eq.) in a mixture of THF (2.00 ml), MeOH (2.00 ml) and H ₂ O (1.00 ml) added LiOH-H ₂ O (369.03 mg, 8.79 mmol, 5.00 eq.) at 0°C. The reaction mixture was slowly warmed to room temperature, stirred continuously for 1 hour, and then adjusted to approximately pH=5 with 1N dilute hydrochloric acid. The resulting mixture solution was concentrated and extracted with ethyl acetate. The organic phase was combined and concentrated to give the crude product of compound 4-8, which was used immediately in the next step. MS (ESI) m/z: 270.9 [M+1].

Step 7: Synthesis of Compound 4-9

To a solution of compound 4-8 (0.5 g, 1.85 mmol, 1.00 eq.) in DMF (8.00 ml) was added compound 3-7 (841.99 mg, 2.22 mmol, 1.20 eq.), TBTU (712.90 mg, 2.22 mmol, 1.20 eq.) and DIEA (526.08 mg, 4.07 mmol, 2.20 eq.) at -10°C. The reaction mixture was stirred at -10°C to 0°C for 0.5 h, and then 10 ml of water was added thereto. The resulting mixture was extracted with ethyl acetate. The organic phase was combined and washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The resulting crude product was purified by silica gel column chromatography to give compound 4-9. MS (ESI) m/z: 518.2 [M+1].

Step 8: Synthesis of Compound WX-407

Isobutylboronic acid (579.23 mg , 5.68 mmol, 7.00 eq.) and HCl (1 M, 1.62 ml, 2.00 eq.) at 0°C. The reaction mixture was slowly warmed to room temperature and stirred continuously for 12 hours, and then 5.00 ml of n-hexane was added thereto. The resulting mixture solution was extracted with MeOH (10 ml). The methanol layer was adjusted to pH 5-6 with saturated sodium bicarbonate aqueous solution and then extracted with ethyl acetate. The organic phase was combined and dried over anhydrous sodium sulfate, filtered and concentrated. The resulting crude product was purified by preparative HPLC to give compound WX-407. MS (ESI) m/z: 366.1 [M-17].

1Н NMR (400 MHz, METHANOL-d ₄ ) δ 6.96 (ddd, J=5.02, 8.91, 12.17 Hz, 1H), 6.40-6.58 (m, 1H), 6.34

- 54 040139 (ddd, J=3.14, 7.15, 10.04 Hz, 1H), 4.49-4.70 (m, 2H), 4.12 (s, 3H), 3.78-4.00 (m, 1H), 2.57-2.79 (m, 2H ), 2.402.53 (m, 1H), 1.57-1.76 (m, 1H), 1.25-1.46 (m, 2H), 0.81-1.01 (m, 6H).

Method for Preparative HPLC Separation of Compound WX-407:

Column: Xtimate C18 150x25 mmx5 µm

Mobile phase: A: water (0.225% FA); B: MeOH, elution gradient B%: 59%-89%

The retention time of WX-407 was 9.5 minutes using the high performance liquid column chromatography method.

Experimental Example 1 In Vitro Antiproliferative Activity Test on MM1.S Cells

This test aims to investigate the proliferation inhibitory effect of a compound by determining the effect of the compound on cell viability of the MM1.S tumor cell line in vitro.

MM1.S cells were seeded in a black 96-well cell culture plate at a density of 7000 cells per well, and then the plate was incubated overnight in a 5% CO ₂ and 100% relative humidity incubator at 37°C. A solution of the test compound in DMSO was added to the cell culture wells at a certain concentration (from 0.3 to 2000 nM) and then the culture plate was placed back into the incubator and provided with a vehicle control (containing DMSO without compound) and a blank control.

The culture plate was incubated in an incubator with 5% CO ₂ and 100% relative humidity at 37°C for 2 days. The sample was processed in a standard manner using the Promega CellTiter-Glo Luminescent Cell Viability Assay Kit (Promega-G7571) and the luminescence signal was detected on a SpectraMax i3x plate reader from Molecular Devices. The degree of inhibition of the test compound was calculated by the following equation using the original data:

_ή . „ RLU control with carrier medium - RLU compounds , % degree of inhibition = -------------------------------------------------- ------------χ 100%.

RLU of control with carrier medium - RLU of empty

RLU is the relative luminescence intensity. The result of the in vitro test of the antiproliferative activity of the test compound against MM1.S cells is shown in Table 1. 1.

____________________________________________________Table 1

Compound WX-174 WX-193 WX-333 WX-260 WX-301 WX-306 WX-308 WX-313 WX-317 WX-327 WX-329 WX-351 WX-355 WX-365 WX-367 WX-373 WX-379 WX-381 WX-385 WX-391

IC50 (µM) 0.0058 0.0082 0.0010 0.0110 0.0058 0.0082 0.0089 0.0245 0.0320 0.0242 0.0115 0.0459 0.0288 0.0906 0.1831 0.0267 0.0240 0.1194 0.1045 0.0054

Experimental Example 2 Stability of Test Compounds in Liver Microsome

The test compound was incubated with CD-I mouse liver microsome, SD rat liver microsome and human liver microsome, respectively, to evaluate the stability of the test compound.

Preparation of test compound solution sample: A 10 mM solution (5 µl) of the compound prepared in Example Compound in DMSO was added to a mixed solvent (450 µl) of DMSO (45 µl), methanol and water (the volume ratio of methanol to water was 1:1) with obtaining a 100 μM solution of the test compound; and 50 μl of 100 μM test compound solution was added to 450 μl of 100 mM potassium phosphate buffer to prepare a 10 μM test compound solution.

- 55 040139 μM solution of the test compound was pre-incubated with the microsome from the above three species for 10 minutes, respectively, and then the working solution of the recovery system based on reduced nicotinamide adenine dinucleotide phosphate (NADPH) was added to the incubation plate at each time point to initiate the reaction, and finally , stop solution (100% ACN) was added to the reaction plate to terminate the reaction after 0, 5, 10, 20, 30 and 60 minutes. The test compound was determined by the LC-MS/MS method. The result of the stability test of the test compound in the liver microsome is shown in table. 2.

table 2

Compound Stability in the liver microsome (T1/2, minutes) WX-174 3.5(H), 1.6(R), 0.8(M) WX-193 4.8(H), 4.6(R), 2.7(M) WX-333 67.4(H), 43.1(R), 67.4(M) WX-260 107.9(H), >145(R), 84.8(M) WX-301 74.3(H), 42.1(R), 43.5(M) WX-306 17.8(H), 37.4(R), 14.0(M) WX-308 27.4(H), 26.8(R), 20.1(M) WX-313 >145(H), 57.2(R), 52.5(M) WX-317 77.0(H), 35.5(R), 27.1(M) WX-327 75.0(H), 26.5(R), 37.7(M) WX-329 51.5(H), 43.3(R), 40.3(M)

Note: H represents a person

R is a rat and M is a mouse.

Experimental Example 3 Cell Membrane Permeability of Test Compounds The cell membrane permeability of a test compound was evaluated on MDR1-MDCKII cells.

The test compound (10 mM solution of the compound in DMSO) was diluted with transfer buffer (HBSS with 10 mM Hepes, pH = 7.4) to obtain a sample having a final concentration of 2 μM, followed by bidirectional (A-B and B-A) administration . After injection, the cell plate was incubated in an incubator with 5% CO2 and saturated humidity at 37°C for 150 min. After a 150 minute incubation, a sample was collected and the concentration of test compound in the transfer sample was determined semi-quantitatively using the LC/MS/MS method. The test result of the permeability of the test compound through the cell membrane is shown in table. 3.

Table 3

Compound Papp A - B (10e-6 cm/s) Papp B - A (10e-6 cm/s) Efflux coefficient WX-174 0.44 18.39 41.79 WX-193 0.10 13.06 130.60 WX-333 0.38 7.32 19.26 WX-260 0.30 5.10 17.00 WX-306 0.30 17.80 59.33 WX-308 0.20 15.30 76.50 WX-313 0.20 1.29 7.16 WX-317 0.01 0.05 5.00 WX-327 0.24 2.24 9.33 WX-329 0.27 3.74 13.85

Note: Papp A - B represents the rate at which the compound enters the cell; Papp B - A is the rate at which the cell removes the compound;

efflux coefficient \u003d Papp B - A / Papp A - B.

Claims (20)

CLAIM 1. A compound of formula (I), a pharmaceutically acceptable salt or stereoisomer thereof - 56 040139 where ring A is selected from the group consisting of C _3-6 cycloalkyl, phenyl and 5-10 membered heteroaryl, where the prefix hetero in 5-10 membered heteroaryl is each independently selected from the group consisting of -O-, -S-, -NH- and N; and the number of heteroatoms or a group containing a heteroatom is independently selected from 1, 2 or 3; n is selected from 0, 1,2 or 3; R1 is each independently selected from the group consisting of halo, OH, NH2, CN, C _1-3 alkyl, C _1-3 alkoxy, and phenyl, where C _1-3 alkyl, C _1-3 alkoxy, or phenyl are each unsubstituted or substituted with 1 , 2 or 3R; R3 is H; R2 is selected from the group consisting of H, halo, OH, NH2, CN, C _1-6 alkyl, C _1-6 alkoxy, C _1-3 alkylO-Ci-3 alkyl-, C _3-6 cycloalkyl-(CH ₂ ) _1-3 - and phenyl-(CH ₂ ) _1-3 -, where C _1-6 alkyl, C _1-6 alkoxy, C _1-3 αkyl-OC _1-3 alkyl-, C _3-6 cycloalkyl-( CH ₂ ) _1-3 - or phenyl-(CH ₂ ) _1-3 - each unsubstituted or substituted with 1, 2 or 3 R; or R2 and R3 together with the carbon atom to which they are attached form a 3-6 membered cycloalkyl; R ₄ is selected from the group consisting of C _1-6 alkyl and C _3-6 cycloalkyl-(CH ₂ ) _1-3 -; R5 is selected from the group consisting of H and C _1-3 alkyl; each R is independently selected from the group consisting of F, Cl, Br, I, OH, Me, NH2, NH( _CH3 ) and N(CH3)2. 2. The connection according to claim 1, where n is selected from 0, 1 or 2. 3. A compound according to claim 1 or 2, wherein R ₁ is each independently selected from the group consisting of halo, OH, NH2, CN, C _1-3 alkoxy, phenyl, and C _1-3 alkyl, unsubstituted or substituted with 1, 2, or 3F. 4. The compound of claim 3 wherein R1 is each independently selected from the group consisting of F, Cl, CN, CF3 and 5. A compound according to any one of claims 1 to 4, wherein ring A is selected from the group consisting of cyclopropyl, phenyl, 5-membered heteroaryl, 6-membered heteroaryl and 9-membered heteroaryl. 6. A compound according to claim 5 wherein ring A is selected from the group consisting of cyclopropyl, phenyl, pyridyl, pyrimidinyl, pyrazinyl and thiazolyl. η 7 A) 7. The compound according to claim 1, where the AC unit in the compound of formula (I) is selected from the group consisting of 8. Connection by item 7, where the structural unit in the compound of formula (I) is selected from the group consisting of - 57 040139 9. Connection according to any one of claims 1 to 8, where R ₃ represents H; R2 is selected from the group consisting of H, C1-6 alkyl, C _1-3 alkoxy, C _1-3 alkyl-OC ₁ . ₃ alkyl-, C _3-6 cycloalkyl-CH ₂ - and phenyl-CH ₂ -, where C _1-6 alkyl, C1.3 alkoxy, C _1-3 alkyl-OC ₁ . ₃ alkyl-, C _3-6 cycloalkyl-CH ₂ - or phenyl-CH ₂ - each unsubstituted or substituted with 1, 2 or 3 substituents independently selected from the group consisting of F, Cl, Br, I and hydroxyl. 10. A compound according to claim 9, wherein R ₃ is H and R 2 is selected from the group consisting of H, Me, 11. A compound according to any one of claims 1-10, wherein R4 is selected from the group consisting of C _3-4 alkyl and C _3-4 cycloalkyl-CH ₂ -. 12. The compound according to claim 11, where R4 is selected from 13. A compound according to any one of claims 1-12, wherein R5 is selected from the group consisting of H and Me. 14. A compound according to any one of claims 1 to 13, wherein each R is independently selected from the group consisting of F and OH. 15. A compound according to any one of claims 1 to 14, wherein the compound of formula (I) is selected from the group consisting of a compound of formula (II), a compound of formula (III), a compound of formula (IV), a compound of formula (V), a compound of formula (VI), compounds of formula (I-a), compounds of formula (I-b), compounds of formula (II-a), compounds of formula (II-b), compounds of formula (III-a), compounds of formula (III-b), compounds of formula (IV-a), compounds of formula (IV-b), compounds of formula (V-a), compounds of formula (Vb), compounds of formula (VI-a) and compounds of formula (VI-b), - 58 040139 - 59 040139 where ring C is selected from the group consisting of cyclopropyl, 5-membered heteroaryl and 6-membered heteroaryl. in formula (IV-a), formula (IV-b), 16. The compound according to claim 15, where the structural unit of formula (VI-a) or formula (VI-b) is selected from the group consisting of 17. A compound according to any one of claims 1 to 16, wherein the compound of formula (I) is selected from the group consisting of - 60 040139 - 61 040139 from or a pharmaceutically acceptable salt or stereoisomer thereof. 18. A compound according to any one of claims 1 to 17, wherein the compound of formula (I) is selected from the group consisting of - 62 040139 - 63 040139 - 64 040139 - 65 040139 or a pharmaceutically acceptable salt thereof. 19. A pharmaceutical composition containing a compound according to any one of claims 1 to 18, a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier, excipient or adjuvant. 20. A method for preventing or treating multiple myeloma, the method comprising administering to a subject in need thereof a compound of any one of claims 1 to 18, a pharmaceutically acceptable salt or stereoisomer thereof.