CN116655605A

CN116655605A - A kind of phenylpyrazole compound, its preparation method and application

Info

Publication number: CN116655605A
Application number: CN202310194497.4A
Authority: CN
Inventors: 王任小; 李嫣; 龚奇能; 周宓; 李晴; 王浩杰
Original assignee: Fudan University
Current assignee: Fudan University
Priority date: 2023-03-02
Filing date: 2023-03-02
Publication date: 2023-08-29
Anticipated expiration: 2043-03-02
Also published as: CN116655605B

Abstract

本发明公开了一种苯基吡唑类化合物、及其制备方法和应用。本发明提供了一种如式I所示的苯基吡唑类化合物、其立体异构体或其药学上可接受的盐。该类化合物能够在分子水平上有效地、选择性地抑制细胞凋亡过程中的关键蛋白MCL‑1，或者，它们对癌细胞，尤其是人浆细胞白血病细胞H929和人髓性单核细胞白血病细胞MV‑4‑11具有明显的杀伤作用和高选择性，具有制备成为新型抗急性髓性白血病药物的潜力，具有较好的市场化前景。 The invention discloses a phenylpyrazole compound, its preparation method and application. The present invention provides a phenylpyrazole compound represented by formula I, its stereoisomer or a pharmaceutically acceptable salt thereof. Such compounds can effectively and selectively inhibit the key protein MCL-1 in the process of apoptosis at the molecular level, or they can inhibit cancer cells, especially human plasma cell leukemia cells H929 and human myelomonocytic leukemia cells The cell MV‑4‑11 has obvious killing effect and high selectivity, has the potential to be prepared as a new type of anti-acute myeloid leukemia drug, and has a good market prospect.

Description

Phenylpyrazole compound, and preparation method and application thereof

Technical Field

The invention discloses a phenylpyrazole compound, and a preparation method and application thereof.

Background

Apoptosis and cell proliferation are fundamental phenomena of life. Modern research suggests that tumor development is actually caused by imbalance in proliferation and apoptosis of cells. Therefore, restoring the intrinsic apoptotic mechanisms in tumor cells is a well-established scientific and effective antitumor strategy. As a key regulatory factor in the mitochondrial apoptosis pathway, the development of small molecule drugs targeting BCL-2 family proteins has become a hotspot for anti-tumor research. Successful marketing of valnemulin verifies this target and its tremendous market potential.

However, as a selective inhibitor of BCL-2 protein, the pro-apoptotic factors released by the binding of vitamin netrop to BCL-2 protein can still be captured by another member of the family, MCL-1 protein, thus inhibiting activation of pro-apoptotic factors, causing resistance problems in MCL-1 protein over-expressing tumor cells (e.g. leukemia, multiple myeloma, etc.). In addition, a great deal of research shows that MCL-1 is one of the main driving carcinogen factors in the occurrence process of the acute myelogenous leukemia, is a potential target for treating the acute myelogenous leukemia, and has great market demand and development potential. Therefore, the development of selective small molecule inhibitors targeting the MCL-1 protein can effectively solve the drug resistance problem of the existing targeted BCL-2 drugs, expand the application range of the targeted BCL-2 drugs for resisting tumors, and provide a new treatment strategy for malignant tumors mainly driven by the MCL-1 protein.

Disclosure of Invention

The invention aims to solve the technical problems of high selectivity and high inhibition activity of MCL-1 small molecule inhibitor or lack of protease degradation agent, and provides a phenylpyrazole compound, a pharmaceutical composition, a preparation method and application thereof. The compound can effectively and selectively inhibit the key protein MCL-1 in the apoptosis process at the molecular level. Or the derivatives have obvious killing effect and high selectivity on cancer cells, especially human plasma cell leukemia cell H929 and human myelomonocytic leukemia cell MV-4-11, have the potential of being prepared into novel acute myelogenous leukemia resisting medicines, and have good market prospect.

The invention provides a phenylpyrazole compound shown in a formula I, a stereoisomer or a pharmaceutically acceptable salt thereof,

wherein,,

R ^a is R ¹ -(CH ₂ ) _m -；

M is 0, 1,2,3, 4, 5 or 6;

said R is ¹ Is phenyl or is substituted by one or more R ^1-1 A substituted phenyl group; when a plurality of substituents are present, the substituents are the same or different;

said R is ^1-1 independently-NH ₂ 、-OH、-SH、C ₁ -C ₄ Alkoxy, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl; the number of heteroatoms in the 3-8 membered heterocycloalkyl is 1,2,3 or 4, and the heteroatoms are N, O and S independently;

R ^b1 、R ^b2 and R is ^b3 Independently H, halogen or C ₁ ～C ₆ An alkyl group;

R ^c is H or C ₁ ～C ₄ An alkyl group;

R ^d is phenyl, 3-10 membered heteroaryl, substituted with one or more R ² Substituted phenyl or by one or more R ² Substituted 3-10 membered heteroaryl; the "3-10 membered heteroaryl" and "are substituted with one or more R ² In the 3-10 membered heteroaryl group in the substituted 3-10 membered heteroaryl group ", the number of heteroatoms is independently 1,2,3 or 4, and the heteroatoms are independently N, O and S; when a plurality of substituents are present, the substituents are the same or different;

the R is ² Independently hydroxyOr C ₁ ～C ₆ An alkyl group;

indicating z configuration->E configuration->Or a mixture thereof;

the chiral carbon labeled ". X" is in S configuration, R configuration, or a mixture thereof.

In one embodiment, R is ¹ Is R ^1-1 A substituted phenyl group.

In one embodiment, R is ^1-1 independently-NH ₂ 、-OH、C ₁ -C ₄ Alkoxy, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl; the number of heteroatoms in the 3-8 membered heterocycloalkyl group is 1,2,3 or 4, and the heteroatoms are N, O and S independently.

In one embodiment, R is ^1-1 independently-NH ₂ -OH, -SH or 3-8 membered cycloalkyl.

In one embodiment, R is ^1-1 independently-NH ₂ -OH, or 3-8 membered cycloalkyl.

In one embodiment, R is ^1-1 Independently C ₁ -C ₄ Alkoxy or 3-8 membered heterocycloalkyl; the number of heteroatoms in the 3-8 membered heterocycloalkyl group is 1,2,3 or 4, and the heteroatoms are N, O and S independently.

In one embodiment, R is ^b1 、R ^b2 And R is ^b3 One of which is halogen or C ₁ ～C ₆ Alkyl, the rest is H; preferably, R ^b2 Is halogen or C ₁ ～C ₆ Alkyl, R ^b1 And R is ^b3 Is H; more preferably, R ^b2 Is halogen, R ^b1 And R is ^b3 H.

In one embodiment, R is ^d Is 3-10 membered heteroarylIs/are R ² Substituted phenyl or by one or more R ² Substituted 3-10 membered heteroaryl; the "3-10 membered heteroaryl" and "are substituted with one or more R ² In the 3-10 membered heteroaryl group in the substituted 3-10 membered heteroaryl group ", the number of hetero atoms is 1,2,3 or 4, and the hetero atoms are N, O and S independently.

In one embodiment, R is ^d Is 3-10 membered heteroaryl; the number of heteroatoms in the 3-10 membered heteroaryl is 1,2,3 or 4, and the heteroatoms are N, O and S independently.

In one embodiment, R is ² Is C ₁ ～C ₆ An alkyl group.

In one embodiment, wherein

R ^a Is R ¹ -(CH ₂ ) _m -；

M is 0;

said R is ¹ Is R ^1-1 A substituted phenyl group;

said R is ^1-1 independently-NH ₂ -OH, or 3-8 membered cycloalkyl;

R ^c is H or C ₁ ～C ₄ An alkyl group;

R ^d is 3-10 membered heteroaryl or "substituted with one or more R ² Substituted 3-10 membered heteroaryl "; the "3-10 membered heteroaryl" and "are substituted with one or more R ² In the 3-10 membered heteroaryl group in the substituted 3-10 membered heteroaryl group ", the number of hetero atoms is 1 or 2, and the hetero atoms are N;

the R is ² Is C ₁ ～C ₆ An alkyl group.

In one embodiment, wherein

R ^a Is R ¹ -(CH ₂ ) _m -；

M is 0;

said R is ¹ Is R ^1-1 A substituted phenyl group;

said R is ^1-1 Independently C ₁ -C ₄ Alkoxy or 3-8 membered heterocycloalkyl; the number of heteroatoms in the 3-8 membered heterocycloalkyl is 1,2,3 or 4, and the heteroatoms are N, O and S independently;

R ^c is H or C ₁ ～C ₄ An alkyl group;

the R is ² Is C ₁ ～C ₆ An alkyl group.

In one embodiment, m is 0, 1, 2, 3, or 4.

In one embodiment, m is 0.

In one embodiment, R is ^1-1 The number of substitutions may be one or more, for example 2, 3, 4 or 5, when a plurality of R's are present ^1-1 When R is ^1-1 The same or different.

In one embodiment, R is ^1-1 May be independently located ortho, meta or para to the "phenyl and pyrazole attachment site", preferably para.

In one embodiment, when R ^1-1 Is C ₁ -C ₄ Alkoxy, the C ₁ -C ₄ Alkoxy is methoxy, ethoxy, propoxy or butoxy, preferably methoxy.

In one embodiment, when R ^1-1 In the case of a 3-8 membered cycloalkyl group, the 3-8 membered cycloalkyl group is cyclopropyl, cyclopentyl, cyclohexyl or cycloheptyl, preferably cyclohexyl.

In one embodiment, when R ^1-1 In the case of 3-8 membered heterocycloalkyl, the 3-8 membered heterocycloalkyl is a 5-6 membered heterocycloalkyl having 1 to 2 heteroatoms selected from N and/or O, for example morpholinyl.

In one embodiment, when R ^b1 、R ^b2 And R is ^b3 When independently halogen, the halogen is fluorine, chlorine, bromine or iodine, preferably chlorine.

In one embodiment, when R ^b1 、R ^b2 And R is ^b3 Independently C ₁ ～C ₆ When alkyl, the C ₁ ～C ₆ Alkyl is preferably C ₁ ～C ₄ Alkyl groups such as methyl, ethyl, propyl or butyl.

In one embodiment, when R ^c Is C ₁ ～C ₄ When alkyl, the C ₁ ～C ₄ Alkyl is methyl, ethyl, propyl or butyl.

In one embodiment, when R ^d Is "3-10 membered heteroaryl" or "is substituted with one or more R ² In the case of a substituted 3-10 membered heteroaryl group ", the 3-10 membered heteroaryl group is a 5-10 membered heteroaryl group having 1 to 2 heteroatoms, such as indolyl, selected from N.

In one embodiment, when R ^d To be covered by one or more R ² When substituted 3-10 membered heteroaryl, when multiple R's are present ² When R is ² The same or different.

In one embodiment, when R ^d To be covered by one or more R ² R in the case of substituted 3-10 membered heteroaryl ² Independently located ortho, meta or para to the attachment site to the other group;

in one embodiment, when R ² Is C ₁ ～C ₆ When alkyl, the C ₁ ～C ₆ Alkyl is C ₁ ～C ₄ Alkyl groups such as methyl, ethyl, propyl or butyl.

In one embodiment, the R ^a Is that

In one embodiment, the R ^d Is that

In one embodiment, R is ^1-1 independently-NH ₂ 、-OH、C ₁ -C ₄ Alkoxy, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl; the 3-8 membered heterocycloalkyl is 5-6 membered heterocycloalkyl with heteroatom number of 1-2, and the heteroatom is selected from N and/or O.

In one embodiment, R is ^1-1 independently-NH ₂ 、-OH、C ₁ -C ₄ Alkoxy, 3-8 membered cycloalkyl or morpholinyl.

In one embodiment, R is ^d Is phenyl, 3-10 membered heteroaryl, substituted with one or more R ² Substituted phenyl or by one or more R ² Substituted 3-10 membered heteroaryl; the "3-10 membered heteroaryl" and "are substituted with one or more R ² In the 3-10 membered heteroaryl group in the substituted 3-10 membered heteroaryl group ", the number of hetero atoms is 1 or 2, and the hetero atoms are N.

In one embodiment, R is ^d Is 3-10 membered heteroaryl or is substituted with one or more R ² Substituted 3-10 membered heteroaryl; the "3-10 membered heteroaryl" and "are substituted with one or more R ² In the 3-10 membered heteroaryl group in the substituted 3-10 membered heteroaryl group ", the number of hetero atoms is 1 or 2, and the hetero atoms are N.

In one embodiment, R is ^d Is phenyl, indolyl or is covered with oneOr a plurality of R ² Substituted indolyl.

In one embodiment, R is ^d Is indolyl or is substituted by one or more R ² Substituted indolyl.

In one embodiment, R is ² Independently is hydroxy or C ₁ ～C ₄ An alkyl group.

In one embodiment, the phenylpyrazole compound I may be any of the following compounds:

the invention provides a compound shown in a formula II, a stereoisomer or a pharmaceutically acceptable salt thereof,

wherein,,

x is a ligand of E3 (ubiquitin) ligase, which is a binder

L is a linker (1 linker), e.g.

The end a is connected with Y, and the end b is connected with X;

n1 and n2 are independently 0, 1,2,3, 4 or 5;

L ¹ and L ² Independently is-O- -S-or-NH-;

y is(target protein conjugate binder);

R ^b1 、R ^b2 、R ^b3 、R ^c and R is ^d Is defined as in any one of the above schemes;

L ³ is R ³ -(CH ₂ ) _m -；

M is 0, 1,2,3, 4, 5 or 6;

said R is ³ Is phenyl or R ^3-1 A substituted phenyl group;

the L is connected to L ³ Ortho, meta or para to the pyrazole attachment site;

the R is ^3-1 independently-NH ₂ 、-OH、-SH、C ₁ -C ₄ Alkoxy, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl; the number of heteroatoms in the 3-8 membered heterocycloalkyl is 1,2,3 or 4, and the heteroatoms are N, O and S independently;

represents Z configuration, E configuration or a mixture thereof;

In one embodiment of the invention, L is

In one aspect of the invention, the L ¹ is-O-.

In one aspect of the invention, the L ² is-O-.

In one aspect of the invention, the L is attached to L ³ Para to the pyrazole attachment site.

In one embodiment of the invention, R is ³ Is phenyl.

In one aspect of the invention, the R ^3-1 independently-NH ₂ 、-OH、C ₁ -C ₄ Alkoxy, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl; the hetero atom in the 3-8 membered heterocyclic alkyl group isThe number is 1,2,3 or 4, and the heteroatoms are independently N, O and S.

In one embodiment, R is ^3-1 independently-NH ₂ -OH, -SH or 3-8 membered cycloalkyl.

In one embodiment, R is ^3-1 independently-NH ₂ -OH, or 3-8 membered cycloalkyl.

In one embodiment, R is ^3-1 Independently C ₁ -C ₄ Alkoxy or 3-8 membered heterocycloalkyl.

In one embodiment, n1 is 0, 1,2 or 3.

In one embodiment, n2 is 0, 1,2 or 3.

In one embodiment, said m is 0, 1,2,3 or 4.

In one embodiment, m is 0.

In one embodiment, R is ^3-1 The number of substitutions may be one or more, for example 2,3, 4 or 5, when a plurality of R's are present ^3-1 When R is ^3-1 The same or different.

In one embodiment, when R ^3-1 Is C ₁ -C ₄ Alkoxy, the C ₁ -C ₄ Alkoxy is methoxy, ethoxy, propoxy or butoxy, preferably methoxy.

In one embodiment, when R ^3-1 In the case of a 3-8 membered cycloalkyl group, the 3-8 membered cycloalkyl group is cyclopropyl, cyclopentyl, cyclohexyl or cycloheptyl, preferably cyclohexyl.

In one embodiment, when R ^3-1 In the case of 3-8 membered heterocycloalkyl, the 3-8 membered heterocycloalkyl is a 5-6 membered heterocycloalkyl having 1 to 2 heteroatoms selected from N and/or O, for example morpholinyl.

In one embodiment, R is ^3-1 Independently of the NH ₂ 、-OH、C ₁ -C ₄ Alkoxy, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl; the 3-8 membered heterocycloalkyl is 5-6 membered heterocycloalkyl with heteroatom number of 1-2, and the heteroatom is selected from N and/or O.

At a certain sideIn the scheme, R is ^3-1 independently-NH ₂ 、-OH、C ₁ -C ₄ Alkoxy, 3-8 membered cycloalkyl or morpholinyl.

In one embodiment, X is

In one embodiment, L is

In one embodiment, Y is

In one embodiment of the present invention, the compound of formula II, its stereoisomer or its pharmaceutically acceptable salt may be a compound of formula II-a as follows

Wherein R is ^c 、R ^b1 、R ^b2 、R ^b3 And R is ^d Is defined as above;

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

In one embodiment of the present invention, the compound of formula II, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof may be any one of the following compounds:

the invention also provides a preparation method of the compound shown in the formula I, which comprises a method 1 or a method 2;

the method 1 comprises the following steps: in an organic solvent, carrying out amidation reaction on the compound III and the compound IV in the presence of a catalyst to obtain the compound I;

wherein R is ^a 、R ^b1 、R ^b2 、R ^b3 And R is ^d Is defined as above.

The method 2 comprises the following steps: in organic solvent, "R" is used ^c Is C ₁ ～C ₄ The compound I of the alkyl group "is subjected to hydrolysis reaction as shown below to give" R " ^c The compound I is-OH';

wherein R is ^a 、R ^b1 、R ^b2 、R ^b3 And R is ^d Is defined as above.

In method 1, the conditions for the amidation reaction may be conventional conditions for the reaction in the art, for example, the following conditions:

the organic solvent may be an amide-based solvent (e.g., N-dimethylformamide). The amount of the organic solvent may be an amount conventionally used in the art so as not to affect the reaction.

The catalyst may be one or more of N, N ' -Diisopropylethylamine (DIPEA), 2- (7-benzotriazol-oxide) -N, N, N ', N ' -tetramethyluronium Hexafluorophosphate (HATU) and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI).

The molar ratio of said compound III to said compound IV is preferably 1:1 to 1:2 (e.g. 1:2).

The progress of the amidation reaction may be monitored by conventional monitoring methods in the art (e.g., TLC, HPLC or NMR), and typically the reaction is terminated when compound III is no longer reacted, and the reaction time may be 6 hours.

In method 2, the conditions of the hydrolysis reaction may be conventional conditions in the art for the reaction, for example, the following conditions:

the organic solvent may be an ether solvent (e.g., tetrahydrofuran). The amount of the organic solvent may be an amount conventionally used in the art so as not to affect the reaction.

The hydrolysis is preferably carried out in the presence of a base, which may be an alkali metal hydroxide (e.g. NaOH).

The preparation method of the compound I can further comprise the following steps: in an organic solvent, in the presence of alkali, performing an addition elimination reaction shown below on the compound IV and the compound V to obtain the compound III;

wherein R is ^a 、R ^b1 、R ^b2 And R is ^b3 As described above;

the R is ^e Is C ₁ ～C ₄ Alkyl (e.g., ethyl).

The conditions for the addition elimination reaction may be those conventional in the art for such reactions, such as the following:

The molar ratio of the compound V to the compound VI is preferably from 1:2 to 1:3 (for example 1:2.2).

The preparation method of the compound I can further comprise the following steps: in an organic solvent, in the presence of a catalyst, carrying out a Wilsmeier-Haack formylation reaction on a compound VII and disubstituted formamide as shown below to obtain the compound V;

wherein R is ^a 、R ^b1 、R ^b2 And R is ^b3 As described above.

The conditions for the Vilsmeier-Haack formylation reaction may be those conventional in the art for such reactions, such as the following:

the organic solvent may be a carboxamide solvent (e.g., N-dimethylformamide). The amount of the organic solvent may be an amount conventionally used in the art so as not to affect the reaction.

The disubstituted formamide can be N, N-dimethylformamide.

The catalyst is POCl ₃ 。

The molar ratio of the compound VII to the catalyst is preferably from 1:2 to 1:5 (for example 1:4).

The preparation method of the compound I can further comprise the following steps: in an organic solvent, in the presence of a catalyst, carrying out condensation reaction of a compound VIII and a compound IX to obtain the compound VII;

Wherein R is ^a 、R ^b1 、R ^b2 And R is ^b3 As described above.

The conditions for the condensation reaction may be those conventional in the art for such reactions, such as the following:

the organic solvent may be an alcoholic solvent (e.g., ethanol). The amount of the organic solvent may be an amount conventionally used in the art so as not to affect the reaction.

The catalyst may be acetic acid.

The molar ratio of compound VIII to compound IX described in the present invention is preferably 1:1 to 1:1.5 (e.g., 1:1.2).

The invention also provides a preparation method of the compound II-a, which comprises the following steps: in a solvent, under the action of a catalyst, reacting the compound II-b with the compound II-c to obtain a compound shown as a formula II-a;

wherein R is ^b1 、R ^b2 、R ^b3 、R ^c And R is ^d Is defined as above;

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

The conditions of the reaction may be those conventional in the art for such reactions, for example, the following conditions:

the solvent may be a formamide solvent (e.g., N-dimethylformamide). The amount of solvent may be that which is conventional in the art so as not to interfere with the reaction.

The catalyst may be N, N' -Diisopropylethylamine (DIPEA).

The molar ratio of said compound II-b to said compound II-c is preferably 1:1 to 1:1.5.

In one embodiment, the method for preparing the compound II-a may further comprise the steps of: removing protecting groups from the compound II-d in a solvent to obtain a compound shown in a formula II-b;

wherein R is ^b1 、R ^b2 、R ^b3 、R ^c And R is ^d Is defined as above;

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

the solvent may be a halogenated hydrocarbon solvent (e.g., methylene chloride). The amount of solvent may be that which is conventional in the art so as not to interfere with the reaction.

The deprotecting reagent may be an acid reagent such as trifluoroacetic acid.

In one embodiment, the method for preparing the compound II-a may further comprise the steps of: in a solvent, reacting the compound II-e with the compound II-f in the presence of alkali and a condensation reagent to obtain a compound shown as a formula II-d;

wherein R is ^b1 、R ^b2 、R ^b3 、R ^c And R is ^d Is defined as above;

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

The catalyst may be N, N ' -Diisopropylethylamine (DIPEA) and 2- (7-oxo-benzotriazol) -N, N, N ', N ' -tetramethylurea Hexafluorophosphate (HATU).

The molar ratio of the compound of formula II-e to the compound II-f is preferably 1:1 to 1:1.5, more preferably 1:1.1 to 1:1.2.

The invention provides a pharmaceutical composition, which comprises a substance A, a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutic adjuvant; the substance A is a compound shown as a formula I or a formula II; the substance a, stereoisomer thereof, or pharmaceutically acceptable salt thereof can be in a therapeutically effective amount.

The invention also provides application of the substance A, a stereoisomer or a pharmaceutically acceptable salt thereof in preparing a Bcl anti-apoptotic protein inhibitor or preparing a medicament for treating and/or preventing diseases related to the Bcl anti-apoptotic protein;

the substance A is the compound I or the compound II;

the "Bcl anti-apoptotic proteins" are, for example, one or more of Bc1-XL, bc1-2 and Mcl-1 proteins, preferably Mcl-1;

the "disease associated with Bcl anti-apoptotic proteins" may be cancer; the cancer is preferably leukemia; such as human acute lymphoblastic leukemia or human plasma cell leukemia.

The invention also provides application of the substance B, the stereoisomer or the pharmaceutically acceptable salt thereof in preparing protease degradation agent; the substance B is a compound II.

The present invention also provides a method of treating a Bcl anti-apoptotic protein related disease comprising administering to a patient a therapeutically effective amount of substance a, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above;

the "Bcl anti-apoptotic proteins" are, for example, one or more of Bcl-XL, bcl-2 and Mcl-1 proteins, preferably Mcl-1;

the "Bcl anti-apoptotic protein related disease" may be cancer; the cancer may be leukemia; such as human acute lymphoblastic leukemia or human plasma cell leukemia.

In such applications, the Bc1 anti-apoptotic protein inhibitor may be used in mammalian organisms; it is also useful in vitro, mainly as an experimental use, for example: the kit can be used as a standard sample or a control sample for comparison or prepared according to a conventional method in the field, and can be used for rapidly detecting the Bcl anti-apoptotic protein inhibition effect.

The pharmaceutical excipients can be those which are widely used in the field of pharmaceutical production. Adjuvants are used primarily to provide a safe, stable and functional pharmaceutical composition, and may also provide means for allowing the subject to dissolve at a desired rate after administration, or for promoting effective absorption of the active ingredient after administration of the composition. The pharmaceutical excipients may be inert fillers or provide a function such as stabilizing the overall pH of the composition or preventing degradation of the active ingredients of the composition. The pharmaceutical excipients can comprise one or more of the following excipients: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, sizing agents, disintegrants, lubricants, anti-adherents, glidants, wetting agents, gelling agents, absorption retarders, dissolution inhibitors, enhancing agents, adsorbents, buffering agents, chelating agents, preservatives, colorants, flavoring agents, and sweeteners.

The pharmaceutical compositions of the present invention may be prepared in accordance with the disclosure using any method known to those of skill in the art. For example, conventional mixing, dissolving, granulating, emulsifying, levigating, encapsulating, entrapping or lyophilizing processes.

The pharmaceutical compositions of the present invention may be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid formulations), inhalation, ocular, rectal, topical or parenteral (infusion, injection, implantation, subcutaneous, intravenous, intra-arterial, intramuscular). The pharmaceutical compositions of the invention may also be in controlled or delayed release dosage forms (e.g., liposomes or microspheres). Examples of solid oral formulations include, but are not limited to, powders, capsules, caplets, soft capsules, and tablets. Examples of liquid formulations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs and solutions. Examples of topical formulations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops or serum formulations. Examples of formulations for parenteral administration include, but are not limited to, solutions for injection, dry formulations which may be dissolved or suspended in a pharmaceutically acceptable carrier, suspensions for injection, and emulsions for injection. Examples of other suitable formulations of the pharmaceutical composition include, but are not limited to, eye drops and other ophthalmic formulations; aerosol: such as nasal sprays or inhalants; a liquid dosage form suitable for parenteral administration; suppositories and lozenges.

Unless otherwise specified, all technical and scientific terms used herein have the standard meaning of the art to which the claimed subject matter belongs. In case there are multiple definitions for a term, the definitions herein control.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention as claimed. As used herein, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Furthermore, the term "comprising" is defined as open and not closed.

Unless otherwise indicated, the present invention employs conventional methods of mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniques or pharmacological detection, and reference is made to procedures and conditions conventional in the art.

The present invention employs, unless otherwise indicated, standard nomenclature of analytical chemistry, organic synthetic chemistry, and medicinal chemistry, and standard laboratory procedures and techniques. In some cases, standard techniques are used for chemical synthesis, chemical analysis, drug preparation, formulation and drug delivery, and treatment of patients.

The term "pharmaceutically acceptable" as used herein is intended to refer to those compounds, materials, compositions and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention prepared from the compounds of the present invention which have the specified substituents found herein with relatively non-toxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts may be obtained by contacting neutral forms of such compounds with a sufficient amount of a base in pure solution or in a suitable inert solvent. When the compounds of the present invention contain relatively basic functional groups, the acid addition salts may be obtained by contacting the neutral form of such compounds with a sufficient amount of an acid in pure solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, and certain specific compounds of the invention contain basic and acidic functionalities and can be converted to either base or acid addition salts. Preferably, the salt is contacted with a base or acid in a conventional manner to isolate the parent compound, thereby regenerating the neutral form of the compound. The parent form of a compound differs from its various salt forms in certain physical properties, such as solubility in polar solvents.

The "pharmaceutically acceptable salts" of the present invention can be synthesized from the parent compound containing an acid or base by conventional chemical methods. In general, the preparation of such salts is as follows: prepared via reaction of these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both. Generally, nonaqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.

The small molecule Bcl anti-apoptotic protein inhibitors of the invention can be used as single agents, or in combination with other therapeutic agents, to enhance the efficacy of these therapeutic agents.

The term "active ingredient", "therapeutic agent", or "active agent" refers to a chemical entity that is effective in treating a disorder, disease, or condition of interest.

The term "comprising" is an open-ended expression, i.e., including what is indicated by the invention, but not excluding other aspects.

In the present invention, the term "C ₁ -C ₆ Alkyl "is preferably each independently methyl, ethyl, propyl, butyl, pentyl or hexyl; wherein propyl is C ₃ Alkyl (including isomers such as n-propyl or isopropyl); butyl is C ₄ Alkyl (including isomers such as n-butyl, sec-butyl, isobutyl, or tert-butyl); pentyl is C ₅ Alkyl (including isomers, e.g. n-pentyl < e.g.) >, isoamyl < e.g.)>> or neopentyl < e.g.)>>; hexyl is C ₆ Alkyl (including isomers, such as n-hexyl);

in the present invention, the term "C ₁ -C ₄ The alkyl groups "are preferably each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl or tert-butyl.

In the present invention, the term "C ₁ -C ₄ Alkoxy "is preferably each independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy or tert-butoxy.

In the present invention, the number of "substitutions" may be one or more < e.g., 2, 3, 4, or 5 >, where there are multiple "substitutions" that are the same or different.

In the present invention, the position of the term "substituted" may be arbitrary unless otherwise specified.

The term "stereoisomer" herein refers to a cis, trans or optical isomer. These stereoisomers may be isolated, purified and enriched by asymmetric synthesis methods or chiral separation methods (including but not limited to thin layer chromatography, rotary chromatography, column chromatography, gas chromatography, high pressure liquid chromatography, etc.), and may be obtained by chiral resolution by bonding (chemical bonding, etc.) or salifying (physical bonding, etc.) other chiral compounds. The term "single stereoisomer" means that one stereoisomer of the compound of the present invention is present in an amount of not less than 95% by mass relative to all stereoisomers of the compound.

The term "aryl" herein refers to a compound having the indicated number of carbon atoms (e.g., C ₆ ～C ₁₀ ) In the case of a monocyclic or polycyclic (e.g., 2 or 3) monovalent hydrocarbon group, which is cyclic, two atoms and one bond are shared between the monocyclic rings, and each ring has aromaticity. Aryl groups are linked through the remainder of the aromatic ring molecule. Aryl groups include, but are not limited to: phenyl and naphthyl.

The term "heteroaryl" as used herein means having the designationA monovalent group of a cyclic, unsaturated, or cyclic type having a specified number of heteroatoms (e.g., 1, 2, or 3) and a specified number of heteroatoms (one or more of N, O and S) and having a cyclic or polycyclic ring, wherein the polycyclic ring is a single ring having two atoms and one bond (condensed) in common, and each ring has aromaticity. Heteroaryl groups are attached to the remainder of the molecule through a carbon atom or heteroatom; heteroaryl groups are attached to the remainder of the molecule through a ring with a heteroatom or a ring without a heteroatom. Heteroaryl groups include, but are not limited to: etc.

The term "heterocycloalkyl" herein refers to a cyclic, saturated monovalent group of a specified number of ring atoms (e.g., 3-8 membered), of a specified number of heteroatoms (e.g., 1, 2, or 3), of a specified class of heteroatoms (one or more of N, O and S), which may be monocyclic, bicyclic, or polycyclic (preferably monocyclic). The heterocycloalkyl group is attached to the remainder of the molecule via a carbon atom or heteroatom. (monocyclic) heterocycloalkyl groups include, but are not limited to: Etc.

The term "cycloalkyl" herein refers to a compound having the indicated number of carbon atoms (e.g., C ₃ ～C ₈ ) A cyclic, saturated monovalent hydrocarbon group which may be monocyclic, bicyclic or polycyclic (preferably monocyclic). Cycloalkyl groups include, but are not limited to:etc.

The term "ligand" is herein a concept in the biological field, referring to a molecule or group that is capable of binding to a protein of interest.

The above preferred conditions can be arbitrarily combined on the basis of not deviating from the common knowledge in the art, and thus, each preferred embodiment of the present invention can be obtained.

The reagents and materials used in the present invention are commercially available.

The invention has the positive progress effects that: the phenylpyrazole compound provided by the invention can effectively and selectively inhibit the key protein MCL-1 in the apoptosis process at the molecular level. Alternatively, they have a pronounced killing effect and high selectivity on cancer cells, in particular human plasma cell leukemia cell H929 and human myelomonocytic leukemia cell MV-4-11. Has the potential of preparing the novel acute myelogenous leukemia resisting medicine and has better market prospect.

Detailed Description

The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.

Example 1 preparation of (E) -1- (1- (2-naphthyl) ethylene) -2- (m-tolyl) hydrazine (1)

To a 100mL eggplant-shaped bottle were added 2-naphthacenedione (3.54 g,20.8 mmol), ethanol (50 mL), acetic acid (0.5 mL), and m-tolylhydrazine hydrochloride (4 g,25 mmol), and the mixture was heated under reflux with stirring for 2h. Water (150 mL) was added and the mixture was extracted 3 times with dichloromethane (30 mL), dried, filtered, and concentrated to the next step.

Example 23 preparation of- (2-Ylnaphthalene) -1- (m-tolyl) -1H-pyrazole-4-carbaldehyde (2)

To a 100mL eggplant-shaped bottle was added (E) -1- (1- (2-naphthyl) ethylene) -2- (m-tolyl) hydrazine (4.93 g,18 mmol), DMF (50 mL), phosphorus oxychloride (5.6 mL,61.9 mmol) was added dropwise at 0deg.C, and the mixture was stirred at room temperature for 6h. Aqueous sodium bicarbonate (150 mL) was added, extracted 3 times with dichloromethane (50 mL), dried, filtered, and concentrated.Petroleum ether ethyl acetate=5:1 column chromatography gives the solid product 3- (2-ylnaphthalene) -1- (m-tolyl) -1H-pyrazole-4-carbaldehyde (2.8 g, 50%). ¹ H NMR(400MHz，CDCl ₃ )δ10.14(s，1H)，8.57(s，1H)，8.33(s，1H)，8.02-7.84(m，4H)，7.68(s，1H)，7.64-7.49(m，3H)，7.40(s，1H)，7.23(s，1H)，2.47(s，3H)。

Example 3 2 preparation of ethyl (3) diphenoxyphosphoryl) acetate

To a 100mL eggplant-shaped bottle was added diphenyl phosphate (8.23 g,43 mmol), anhydrous dichloromethane (20 mL), ethyl bromoacetate (6.2 mL,55.9 mmol), triethylamine (7.77 mL,55.9 mmol) was added dropwise at 0deg.C, and the mixture was stirred at room temperature for 6h. Aqueous solution (100 ml) was added, and dichloromethane (40 ml) was extracted 3 times, dried, filtered, and concentrated. Petroleum ether: ethyl acetate=4:1 column chromatography gives a clear oil (7 g, 51%). ¹ H NMR(400MHz，CDCl ₃ )δ7.39-7.30(m，4H)，7.26-7.12(m，6H)，4.25-4.20(m，2H)，3.35-3.12(m，2H)，1.30-1.26(m，3H)。

Example 4 preparation of (Z/E) -3- (3- (2-Yl-naphthalene) -1- (m-tolyl) -1H-pyrazol-4-yl) acrylic acid Ethyl ester (4)

To a 100mL eggplant-shaped bottle was added ethyl 2- (diphenoxyphosphoryl) acetate (2.8 g,8.8 mmol), anhydrous tetrahydrofuran (20 mL), and a tetrahydrofuran solution (1.25 g,4 mmol) of 3- (2-ylnaphthalene) -1- (m-tolyl) -1H-pyrazole-4-carbaldehyde was added dropwise with stirring, followed by stirring at room temperature for 3 hours. Aqueous solution (100 mL) was added, dichloromethane (30 mL) was extracted 3 times, dried, filtered, concentrated and taken directly to the next step.

Example 5 preparation of (Z/E) -3- (3- (2-naphthyl) -1- (m-tolyl) -1H-pyrazol-4-yl) acrylic acid (5)

A100 mL eggplant-shaped bottle was charged with a mixture of ethyl (Z) -3- (3- (2-ylnaphthalene) -1- (m-tolyl) -1H-pyrazol-4-yl) acrylate and ethyl (E) -3- (3- (2-ylnaphthalene) -1- (m-tolyl) -1H-pyrazol-4-yl) acrylate, tetrahydrofuran (20 mL), and an aqueous sodium hydroxide solution (960 mg,24 mmol) was added dropwise with stirring, followed by heating and stirring for 6 hours. Dilute hydrochloric acid was added to adjust the pH to acidity, extraction was performed 3 times with ethyl acetate (20 mL), drying, filtration, and concentration. Petroleum ether:ethyl acetate=10:1 column chromatography gives cis-5 (0.7 g, 49%) and trans-5 (0.5 g, 35%) as yellow solids. cis-5: ¹ H NMR(600MHz，DMSO-d6)δ12.36(s，1H)，9.24(d，J＝0.6Hz，1H)，8.15(dd，J＝1.6，0.8Hz，1H)，8.07-8.05(m，2H)，8.01-7.99(m，1H)，7.82(dd，J＝8.4，1.7Hz，1H)，7.74(d，J＝2.0Hz，1H)，7.70-7.67(m，1H)，7.60-7.57(m，2H)，7.45(t，J＝7.8Hz，1H)，7.22(ddt，J＝7.5，1.7，0.9Hz，1H)，6.97(dd，J＝12.5，0.7Hz，1H)，5.94(d，J＝12.5Hz，1H)，2.43(s，3H)； ¹³ C NMR(151 MHz，DMSO-d6)δ167.74，153.74，139.92，139.60，133.46，133.29，133.13，131.43，130.06，129.92，128.79，128.73，128.29，128.24，128.10，127.14，127.06，126.89，119.93，118.39，116.57，116.20，21.50；ESI-MS：m/z 355.1[M+H]+，ESI-HRMS：calcd for C ₂₃ H ₁₉ O ₂ N ₂ [M+H] ⁺ 355.1441，found 355.1439.

trans-5： ¹ H NMR(600MHz，DMSO-d6)δ12.26(s，1H)，9.26(s，1H)，8.18(d，J＝1.7Hz，1H)，8.11-8.04(m，2H)，8.01(dd，J＝6.1，3.4Hz，1H)，7.82(d，J＝2.0Hz，1H)，7.80(dd，J＝8.4，1.7Hz，1H)，7.76(dd，J＝8.0，2.2Hz，1H)，7.64-7.58(m，3H)，7.45(t，J＝7.8Hz，1H)，7.22(dd，J＝7.6，1.6Hz，1H)，6.48(d，J＝15.8Hz，1H)，2.43(s，3H)； ¹³ C NMR(151MHz，DMSO-d6)δ168.18，152.45，139.80，139.46，134.65，133.35，133.16，130.04，129.97，129.06，128.89，128.72，128.19，128.17，127.91，127.20，127.18，126.65，119.63，119.12，117.71，116.28，21.53；ESI-MS：m/z 355.1[M+H] ⁺ ，ESI-HRMS：calcd for C ₂₃ H ₁₉ O ₂ N ₂ [M+H] ⁺ 355.1441，found 355.1440.

example 6 preparation of (Z/E) - (3- (3- (2-naphthyl) -1- (m-tolyl) -1H-4-pyrazolyl) acryloyl) tryptophan (I-1, I-2)

To a 50mL eggplant-shaped bottle was added 5 (70 mg,0.2 mmol), anhydrous DMF (10 mL), HATU (114 mg,0.3 mmol), DIPEA (100. Mu.L, 0.6 mmol), L-tryptophan methyl ester hydrochloride (102 mg,0.4 mmol) and stirred at room temperature for 6h. Water (60 mL) was added, extraction 3 times with ethyl acetate (15 mL), drying, filtration, concentration and the next step. 50mL eggplant-shaped bottle was charged with the above raw material, tetrahydrofuran (10 mL), aqueous potassium hydroxide (60 mg), and heated and stirred for 2h. Dilute hydrochloric acid was added to adjust the pH to acidity, extraction was performed 3 times with ethyl acetate (15 mL), drying, filtration, and concentration. Petroleum ether: ethyl acetate=1:1 column chromatography gives yellow solid I-1 (83 mg, 77%), obtained by the same procedure as I-2.

(Z) - (3- (3- (2-naphthyl) -1- (m-tolyl) -1H-4-pyrazolyl) acryloyl) -L-tryptophan (I-1)

¹ H NMR(600MHz，DMSO-d6)δ12.64(s，1H)，10.85(d，J＝2.5Hz，1H)，9.32(s，1H)，8.54(d，J＝7.8Hz，1H)，8.13(d，J＝1.7Hz，1H)，8.07-8.03(m，2H)，7.99(dt，J＝6.9，3.5Hz，1H)，7.79(dd，J＝8.4，1.7Hz，1H)，7.68(d，J＝2.0Hz，1H)，7.62(dd，J＝8.1，2.3Hz，1H)，7.60-7.56(m，3H)，7.42(t，J＝7.8Hz，1H)，7.35-7.31(m，1H)，7.22-7.17(m，2H)，7.07(ddd，J＝8.1，7.0，1.2Hz，1H)，6.99(ddd，J＝7.9，6.9，1.0Hz，1H)，6.70(d，J＝12.5Hz，1H)，6.04(d，J＝12.6Hz，1H)，4.63(ddd，J＝9.2，7.8，4.8Hz，1H)，3.26(dd，J＝14.7，4.8Hz，1H)，3.08(dd，J＝14.7，9.2Hz，1H)，2.42(s，3H)； ¹³ C NMR(151MHz，DMSO-d6)δ174.09，166.12，153.66，139.86，139.65，136.57，133.28，133.07，131.58，130.10，130.03，128.79，128.77，128.65，128.33，128.10，128.06，127.62，127.08，127.06，127.02，124.10，121.42，120.93，119.74，118.87，118.65，116.53，116.47，111.88，110.49，53.57，27.58，21.51；ESI-MS：m/z 541.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₄ H ₂₉ O ₃ N ₄ [M+H] ⁺ 541.2234，found 541.2234.

(E) - (3- (3- (2-naphthyl) -1- (m-tolyl) -1H-4-pyrazolyl) acryloyl) -L-tryptophan (I-2)

¹ H NMR(600MHz，DMSO-d6)δ12.48(s，1H)，10.85(d，J＝2.4Hz，1H)，8.98(s，1H)，8.39(d，J＝7.9Hz，1H)，8.16(d，J＝1.7Hz，1H)，8.09-7.97(m，3H)，7.84-7.74(m，3H)，7.61-7.54(m，3H)，7.48-7.42(m，2H)，7.34(d，J＝8.1Hz，1H)，7.20(d，J＝7.5Hz，1H)，7.16(d，J＝2.4Hz，1H)，7.06(ddd，J＝8.2，6.9，1.2Hz，1H)，6.97(ddd，J＝8.0，6.9，1.0Hz，1H)，6.58(d，J＝15.7Hz，1H)，4.60(ddd，J＝9.2，7.8，4.8Hz，1H)，3.23(dd，J＝14.6，4.8Hz，1H)，3.08(dd，J＝14.7，9.2Hz，1H)，2.42(s，3H)； ¹³ C NMR(151 MHz，DMSO-d6)δ174.04，172.48，165.62，152.08，139.75，139.56，136.58，133.37，133.12，130.27，129.93，129.87，128.80，128.73，128.23，128.14，128.05，127.71，127.62，127.10，126.63，124.05，122.13，121.41，119.70，118.86，118.63，118.16，116.32，111.86，110.51，53.64，27.53，21.54；ESI-MS：m/z 541.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₄ H ₂₉ O ₃ N ₄ [M+H] ⁺ 541.2234，found 541.2235.

Example 7 the esterification product (Z) - (3- (1- (3-chlorophenyl) -3- (4-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester was retained and further hydrolyzed to give the product (Z) - (3- (1- (3-chlorophenyl) -3- (4-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan) by changing 2-naphthacenetone to p-methoxyacetophenone, and changing m-tolylhydrazine hydrochloride to m-chlorophenylhydrazine hydrochloride, and the remaining desired starting materials, reagents, and preparation methods were the same as in example 6. The same procedure gave (E) - (3- (1- (3-chlorophenyl) -3- (4-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester and (E) - (3- (1- (3-chlorophenyl) -3- (4-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan.

(Z) - (3- (1- (3-chlorophenyl) -3- (4-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-31)

¹ H NMR(600MHz，DMSO-d6)δ10.87(d，J＝2.4Hz，1H)，9.21(s，1H)，8.66(d，J＝7.5Hz，1H)，7.89(t，J＝2.1Hz，1H)，7.76(ddd，J＝8.2，2.2，0.9Hz，1H)，7.54(td，J＝7.9，5.7Hz，4H)，7.41(ddd，J＝8.0，2.0，0.9Hz，1H)，7.33(dt，J＝8.1，0.9Hz，1H)，7.18(d，J＝2.2Hz，1H)，7.07(dd，J＝8.8，2.2Hz，3H)，6.99(ddd，J＝7.9，7.0，1.0Hz，1H)，6.58(d，J＝12.6Hz，1H)，6.01(d，J＝12.6Hz，1H)，4.66(ddt，J＝7.4，5.4，3.7Hz，1H)，3.82(s，3H)，3.63(s，3H)，3.22(dd，J＝14.6，5.4Hz，1H)，3.10(dd，J＝14.6，8.8Hz，1H)； ¹³ C NMR(151MHz，DMSO-d6)δ173.13，166.13，160.04，154.00，140.77，136.57，134.53，131.92，131.46，130.52(2C)，128.81，127.49，126.90，124.61，124.22，121.47，120.97，118.92，118.87，118.48，117.62，116.50，114.63(2C)，111.94，109.99，55.71，53.68，52.40，27.55；ESI-MS：m/z 555.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₁ H ₂₈ O ₄ N ₄ Cl[M+H] ⁺ 555.1794，found 555.1792.

(Z) - (3- (1- (3-chlorophenyl) -3- (4-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (I-32)

¹ H NMR(600MHz，DMSO-d6)δ12.71(s，1H)，10.84(d，J＝2.4Hz，1H)，9.26(s，1H)，8.51(d，J＝7.8Hz，1H)，7.88(t，J＝2.1Hz，1H)，7.75(ddd，J＝8.2，2.2，0.9Hz，1H)，7.58-7.51(m，4H)，7.41(ddd，J＝8.0，2.1，0.9Hz，1H)，7.32(d，J＝8.0Hz，1H)，7.17(d，J＝2.1Hz，1H)，7.09-7.04(m，3H)，6.99(ddd，J＝8.0，7.0，1.0Hz，1H)，6.56(d，J＝12.6Hz，1H)，6.01(d，J＝12.6Hz，1H)，4.61(ddt，J＝9.2，4.9，3.1Hz，1H)，3.82(s，3H)，3.25(dd，J＝14.7，4.8Hz，1H)，3.07(dd，J＝14.7，9.1Hz，1H)； ¹³ C NMR(151MHz，DMSO-d6)δ174.05，166.01，160.03，154.05，140.78，136.56，134.54，131.92，131.57，130.54(2C)，128.59，127.62，126.88，124.62，124.10，121.41，121.17，118.90，118.86，118.65，117.58，116.54，114.62(2C)，111.86，110.46，60.23，55.70，27.57；ESI-MS：m/z 541.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₀ H ₂₆ O ₄ N ₄ Cl[M+H] ⁺ 541.1637，found 541.1637.

(E) - (3- (1- (3-chlorophenyl) -3- (4-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-33)

¹ H NMR(600MHz，DMSO-d6)δ10.88(d，J＝2.5Hz，1H)，9.04(d，J＝0.7Hz，1H)，8.58(d，J＝7.6Hz，1H)，8.05(t，J＝2.1Hz，1H)，7.92(ddd，J＝8.3，2.2，0.9Hz，1H)，7.58-7.51(m，4H)，7.42(ddd，J＝8.0，2.0，0.9Hz，1H)，7.38-7.33(m，2H)，7.17(d，J＝2.2Hz，1H)，7.11-7.05(m，3H)，6.99(ddd，J＝8.0，7.0，1.1Hz，1H)，6.52(d，J＝15.7Hz，1H)，4.62(ddt，J＝7.3，5.5，3.7Hz，1H)，3.82(s，3H)，3.62(s，3H)，3.21(dd，J＝14.8，5.2Hz，1H)，3.11(dd，J＝14.7，8.9Hz，1H)； ¹³ C NMR(151MHz，DMSO-d6)δ173.06，165.63，162.78，160.11，152.59，140.71，136.59，134.53，131.81，130.07(2C)，128.39，127.49，126.89，124.71，124.16，121.75，121.47，118.92，118.73，118.46，118.11，117.52，114.76(2C)，111.93，110.07，55.71，53.81，52.35，27.48；ESI-MS：m/z 555.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₁ H ₂₈ O ₄ N ₄ Cl[M+H] ⁺ 555.1794，found 555.1788.

(E) - (3- (1- (3-chlorophenyl) -3- (4-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (I-34)

¹ H NMR(600MHz，DMSO-d6)δ12.69(s，1H)，10.85(d，J＝2.4Hz，1H)，9.02(s，1H)，8.42(d，J＝7.9Hz，1H)，8.04(t，J＝2.1Hz，1H)，7.92(ddd，J＝8.2，2.2，0.9Hz，1H)，7.58-7.53(m，4H)，7.41(ddd，J＝8.0，2.1，0.9Hz，1H)，7.37-7.32(m，2H)，7.16(d，J＝2.1Hz，1H)，7.10-7.05(m，3H)，6.98(ddd，J＝7.9，7.0，1.0Hz，1H)，6.53(d，J＝15.7Hz，1H)，4.59(ddt，J＝9.4，4.9，3.1Hz，1H)，3.82(s，3H)，3.24(dd，J＝14.7，4.8Hz，1H)，3.09(dd，J＝14.7，9.3Hz，1H)； ¹³ C NMR(151 MHz，DMSO-d6)δ174.02，165.54，160.09，152.55，140.72，136.57，134.53，131.80，130.06(2C)，129.79，128.31，127.62，126.86，124.74，124.05，122.14，121.41，118.86，118.72，118.63，118.18，117.51，114.75(2C)，111.86，110.53，60.23，55.71，27.51；ESI-MS：m/z 541.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₀ H ₂₆ O ₄ N ₄ Cl[M+H] ⁺ 541.1637，found 541.1629.

Example 8 the same procedure was followed except for using 2-naphthaleneethanone as the 3-methoxyphenylacetone, m-methylphenylhydrazine hydrochloride as the m-chlorophenylhydrazine hydrochloride, and the remaining desired starting materials, reagents and preparation methods as in example 6, to give (E) - (3- (1- (3-chlorophenyl) -3- (3-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester as the esterification product.

(Z) - (3- (1- (3-chlorophenyl) -3- (3-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-35)

¹ H NMR(600MHz，DMSO-d ₆ )δ10.87(s，1H)，9.21(s，1H)，8.67(d，J＝7.5Hz，1H)，7.91(q，J＝1.9Hz，1H)，7.78(d，J＝8.2Hz，1H)，7.59-7.51(m，2H)，7.47-7.41(m，2H)，7.33(d，J＝8.1Hz，1H)，7.20-7.15(m，2H)，7.14(s，1H)，7.10-7.03(m，2H)，7.00(t，J＝7.5Hz，1H)，6.62(d，J＝12.6Hz，1H)，6.04(d，J＝12.6Hz，1H)，4.69-4.62(m，1H)，3.83(d，J＝1.7Hz，3H)，3.63(s，3H)，3.23(dd，J＝14.7，5.4Hz，1H)，3.10(dd，J＝14.7，8.8Hz，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ172.47，165.47，159.19，153.27，140.09，135.94，133.92，132.97，131.30，130.93，129.65，127.95，126.86，126.44，123.58，120.92，120.84，120.68，118.38，118.29，117.84，117.15，116.12，114.10，113.88，111.30，109.36，55.03，53.06，51.76，26.90.ESI-MS：m/z 555.2[M+H] ⁺ ，

(E) - (3- (1- (3-chlorophenyl) -3- (3-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-36)

¹ H NMR(600MHz，DMSO-d ₆ )δ10.87(d，J＝2.3Hz，1H)，9.06(s，1H)，8.59(d，J＝7.5Hz，1H)，8.06(t，J＝2.0Hz，1H)，7.94(dd，J＝8.2，2.2Hz，1H)，7.58(td，J＝8.1，1.3Hz，1H)，7.52(d，J＝7.9Hz，1H)，7.48-7.33(m，4H)，7.20-7.16(m，3H)，7.10-7.04(m，2H)，6.99(t，J＝7.4Hz，1H)，6.54(dd，J＝15.8，1.3Hz，1H)，4.64(td，J＝8.4，6.3Hz，1H)，3.82(s，3H)，3.62(s，3H)，3.22(dd，J＝14.6，5.5Hz，1H)，3.11(dd，J＝14.6，8.9Hz，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ172.39，164.96，159.28，151.86，140.02，135.96，133.92，133.04，131.20，129.81，129.29，127.95，126.86，126.44，123.50，121.41，120.84，120.47，118.29，118.25，117.82，117.75，117.05，114.13，113.50，111.30，109.42，55.03，53.17，51.71，26.84.ESI-MS：m/z 555.2[M+H] ⁺ ，

Example 9 the same procedure was followed except for using 2-naphthone as the 2-methoxyphenylacetone, m-methylphenylhydrazine hydrochloride as the m-chlorophenylhydrazine hydrochloride, and the remaining desired starting materials, reagents and preparation methods as in example 6, to give (E) - (3- (1- (3-chlorophenyl) -3- (2-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester as the esterification product.

(Z) - (3- (1- (3-chlorophenyl) -3- (2-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-37)

¹ H NMR(600MHz，DMSO-d ₆ )δ10.87(s，1H)，9.34(d，J＝3.0Hz，1H)，8.63-8.59(m，1H)，7.85(s，1H)，7.74(d，J＝8.2Hz，1H)，7.55(tt，J＝8.3，3.0Hz，2H)，7.52-7.46(m，1H)，7.44-7.39(m，1H)，7.367.32(m，2H)，7.19-7.15(m，2H)，7.10-7.04(m，2H)，7.03-6.98(m，1H)，6.21(dd，J＝12.9，3.0Hz，1H)，5.88(dd，J＝12.8，3.0Hz，1H)，4.67(dd，J＝9.2，5.3Hz，1H)，3.75(d，J＝3.0Hz，3H)，3.64(d，J＝3.0Hz，3H)，3.25-3.20(m，1H)，3.13-3.06(m，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ172.51，165.63，156.88，152.58，140.18，135.95，133.90，131.33(2C)，130.48，130.23，128.76，126.86，126.23，123.55，120.84，120.48，120.32，118.90，118.29，118.20，117.84，117.72，116.95，111.49，111.30，109.39，55.23，53.03，51.77，26.91.ESI-MS：m/z 555.2[M+H] ⁺ ，

(E) - (3- (1- (3-chlorophenyl) -3- (2-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-38)

¹ H NMR(600MHz，DMSO-d ₆ )δ10.86(s，1H)，9.00(d，J＝2.7Hz，1H)，8.48(d，J＝7.6Hz，1H)，8.01(s，1H)，7.89(d，J＝8.4Hz，1H)，7.56(td，J＝8.2，2.6Hz，1H)，7.50(t，J＝8.2Hz，2H)，7.41(d，J＝8.1Hz，1H)，7.39-7.32(m，2H)，7.20-7.16(m，1H)，7.15(s，1H)，7.11-7.05(m，3H)，7.01-6.96(m，1H)，6.32(dd，J＝15.8，2.8Hz，1H)，4.59(d，J＝7.6Hz，1H)，3.71(d，J＝2.7Hz，3H)，3.60(d，J＝2.7Hz，3H)，3.223.17(m，1H)，3.123.05(m，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ172.44，165.14，156.77，150.52，140.13，135.93，133.89，131.18，130.99，130.46，130.18，127.08，126.84，126.16，123.46，120.82，120.76，120.36，119.85，119.32，118.28，118.00，117.80，116.80，111.67，111.28，109.47，55.21，53.10，51.67，26.76.ESI-MS：m/z 555.2[M+H] ⁺ ，

Example 10 conversion of 2-naphthaceneethanone to p-methoxyphenylacetone, conversion of m-methylphenylhydrazine hydrochloride to 3, 5-dimethylphenylhydrazine hydrochloride the remaining desired starting materials, reagents and preparation methods are the same as in example 6, leaving the esterified product (Z) - (3- (1- (3, 5-xylyl) -3- (2-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester.

(Z) - (3- (1- (3, 5-xylyl) -3- (2-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-39)

¹ H NMR(400MHz，DMSO-d ₆ )δ10.87(s，1H)，9.17(s，1H)，8.64(d，J＝7.5Hz，1H)，7.52-7.49(m，3H)，7.40(s，2H)，7.32(d，J＝8.0Hz，1H)，7.17(d，J＝2.5Hz，1H)，7.07-7.05(m，3H)，6.99(d，J＝8.8Hz，2H)，6.56(d，J＝12.6Hz，1H)，5.95(d，J＝12.6Hz，1H)，4.63(q，J＝7.6Hz，1H)，3.81(d，J＝2.1Hz，3H)，3.61(s，3H)，3.25-3.16(m，1H)，3.12-3.05(m，1H)，2.34(s，6H).ESI-MS：m/z 549.2[M+H] ⁺ ，

Example 11 the procedure was followed except for using the same materials, reagents and preparation methods as in example 6 except for changing 2-naphthacene to 4' -cyclohexylacetophenone and m-tolylhydrazinium hydrochloride to m-chlorophenylhydrazine hydrochloride, and reserving the esterified product of (Z) - (3- (1- (3- (m-chlorophenyl) -3- (4-cyclohexylphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester, and further hydrolyzing the esterified product to give the product of (Z) - (3- (1- (3- (m-chlorophenyl) -3- (4-cyclohexylphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan.

(Z) - (3- (1- (3- (m-chlorophenyl) -3- (4-cyclohexylphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-40)

¹ H NMR(600MHz，DMSO-d6)δ10.87(d，J＝2.4Hz，1H)，9.22(s，1H)，8.66(d，J＝7.5Hz，1H)，7.89(t，J＝2.1Hz，1H)，7.76(ddd，J＝8.2，2.2，0.9Hz，1H)，7.57-7.50(m，4H)，7.42(ddd，J＝8.1，2.0，0.9Hz，1H)，7.37-7.32(m，3H)，7.18(d，J＝2.2Hz，1H)，7.06(ddd，J＝8.1，6.9，1.2Hz，1H)，6.99(ddd，J＝7.9，7.0，1.0Hz，1H)，6.61(dd，J＝12.6，0.7Hz，1H)，6.02(d，J＝12.6Hz，1H)，4.69-4.62(m，1H)，3.63(s，3H)，3.22(dd，J＝14.6，5.4Hz，1H)，3.10(dd，J＝14.6，8.8Hz，1H)，2.57(tt，J＝11.6，3.3Hz，1H)，1.82(dq，J＝8.9，3.0，2.4Hz，4H)，1.75-1.70(m，1H)，1.50-1.34(m，4H)，1.31-1.21(m，1H)； ¹³ C NMR(151MHz，DMSO-d6)δ173.13，166.13，154.18，148.52，140.76，136.57，134.54，131.94，131.52，129.86，129.19(2C)，128.77，127.49，127.47(2C)，126.96，124.21，121.47，121.06，118.92，118.90，118.47，117.64，116.61，111.94，110.00，53.68，52.40，44.06，34.34(2C)，27.50，26.81(2C)，26.06；ESI-MS：m/z 607.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₆ H ₃₆ O ₃ N ₄ Cl[M+H] ⁺ 607.2470，found 607.2476.

(Z) - (3- (1- (3- (m-chlorophenyl) -3- (4-cyclohexylphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (I-41)

¹ H NMR(600MHz，DMSO-d6)δ12.69(s，1H)，10.84(d，J＝2.4Hz，1H)，9.28(s，1H)，8.52(d，J＝7.8Hz，1H)，7.88(t，J＝2.1Hz，1H)，7.75(ddd，J＝8.2，2.2，0.9Hz，1H)，7.59-7.48(m，4H)，7.42(ddd，J＝8.0，2.1，0.9Hz，1H)，7.38-7.34(m，2H)，7.34-7.31(m，1H)，7.17(d，J＝2.1Hz，1H)，7.06(ddd，J＝8.0，6.9，1.2Hz，1H)，6.98(ddd，J＝8.0，7.0，1.0Hz，1H)，6.58(d，J＝12.6Hz，1H)，6.02(d，J＝12.7Hz，1H)，4.61(ddt，J＝9.2，4.9，3.2Hz，1H)，3.25(dd，J＝14.7，4.8Hz，1H)，3.07(dd，J＝14.7，9.2Hz，1H)，2.57(tt，J＝11.8，3.3Hz，1H)，1.82(td，J＝10.4，9.9，5.1Hz，4H)，1.75-1.69(m，1H)，1.50-1.34(m，4H)，1.31-1.22(m，1H)； ¹³ C NMR(151MHz，DMSO-d6)δ174.04，166.01，154.24，148.51，140.77，136.56，134.54，131.94，131.63，129.87，129.21(2C)，128.56，127.61，127.47(2C)，126.94，124.10，121.41，121.25，118.93，118.86，118.64，117.60，116.65，111.87，110.45，60.23，44.06，34.34(2C)，27.56，26.81(2C)，26.06；ESI-MS：m/z 593.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₅ H ₃₄ O ₃ N ₄ Cl[M+H] ⁺ 593.2314，found 593.2313.

(E) - (3- (1- (3- (m-chlorophenyl) -3- (4-cyclohexylphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-42)

¹ H NMR(600MHz，DMSO-d6)δ10.88(d，J＝2.5Hz，1H)，9.07(s，1H)，8.59(d，J＝7.5Hz，1H)，8.05(t，J＝2.1Hz，1H)，7.95-7.90(m，1H)，7.57(t，J＝8.1Hz，1H)，7.53(dd，J＝8.0，3.5Hz，3H)，7.44-7.41(m，1H)，7.39-7.33(m，4H)，7.17(d，J＝2.1Hz，1H)，7.09-7.05(m，1H)，7.01-6.97(m，1H)，6.54(d，J＝15.7Hz，1H)，4.66-4.59(m，1H)，3.62(s，3H)，3.21(dd，J＝14.6，5.4Hz，1H)，3.11(dd，J＝14.6，9.0Hz，1H)，2.57(tt，J＝11.8，3.3Hz，1H)，1.86-1.79(m，4H)，1.72(dd，J＝13.0，4.0Hz，1H)，1.50-1.34(m，4H)，1.26(dddd，J＝15.9，12.6，8.2，3.7Hz，1H)； ¹³ C NMR(151 MHz，DMSO-d6)δ173.06，165.64，152.84，148.65，140.71，136.59，134.54，131.83，130.05，129.91，128.74(2C)，128.29，127.60(2C)，127.49，126.96，124.15，121.86，121.47，118.92，118.78，118.47，118.25，117.56，111.93，110.07，53.82，52.35，44.06，34.32(2C)，27.47，26.80(2C)，26.05；ESI-MS：m/z 607.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₆ H ₃₆ O ₃ N ₄ Cl[M+H] ⁺ 607.2470，found 607.2478.

(E) - (3- (1- (3- (m-chlorophenyl) -3- (4-cyclohexylphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (I-43)

¹ H NMR(600MHz，DMSO-d6)δ12.67(s，1H)，10.86(s，1H)，9.05(s，1H)，8.44(d，J＝7.8Hz，1H)，8.05(s，1H)，7.93(d，J＝8.3Hz，1H)，7.55(dd，J＝23.6，7.9Hz，4H)，7.44-7.32(m，5H)，7.18(s，1H)，7.07(t，J＝7.5Hz，1H)，6.99(t，J＝7.5Hz，1H)，6.56(d，J＝15.6Hz，1H)，4.62-4.59(m，1H)，3.25(dd，J＝15.1，4.7Hz，1H)，3.10(dd，J＝14.7，9.3Hz，1H)，2.57(t，J＝12.1Hz，1H)，1.82(t，J＝13.8Hz，4H)，1.72(d，J＝13.0Hz，1H)，1.43(dq，J＝25.5，12.7Hz，4H)，1.28-1.21(m，1H)； ¹³ C NMR(151MHz，DMSO-d6)δ174.04，165.56，152.79，148.62，140.72，136.58，134.54，131.81，129.93，129.73，128.72(2C)，128.20，127.63，127.59(2C)，126.93，124.04，122.25，121.41，118.86，118.76，118.64，118.32，117.53，111.86，110.54，53.71，44.07，34.31(2C)，27.52，26.80(2C)，26.05；ESI-MS：m/z 593.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₅ H ₃₄ O ₃ N ₄ Cl[M+H] ⁺ 593.2314，found 593.2323.

Example 12 conversion of 2-naphthacene to propiophenone, conversion of m-methyl phenylhydrazine hydrochloride to m-chlorophenylhydrazine hydrochloride the remaining desired starting materials, reagents and preparation method are the same as in example 6, leaving the esterified product (Z) - (3- (1- (3-chlorophenyl) -3-phenethyl-1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester and further hydrolysis to give the product (Z) - (3- (1- (3-chlorophenyl) -3-phenethyl-1H-pyrazol-4-yl) acryloyl) -L-tryptophan. The same procedure gave (E) - (3- (1- (3-chlorophenyl) -3-phenethyl-1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester and (E) - (3- (1- (3-chlorophenyl) -3-phenethyl-1H-pyrazol-4-yl) acryloyl) -L-tryptophan.

(Z) - (3- (1- (3-chlorophenyl) -3-phenethyl-1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-52)

¹ H NMR(600MHz，Chloroform-d)δ9.11(s，1H)，8.02(s，1H)，7.75(d，J＝2.0Hz，1H)，7.58(dd，J＝8.1，2.1Hz，1H)，7.51(d，J＝7.9Hz，1H)，7.33(t，J＝8.0Hz，1H)，7.31-7.27(m，3H)，7.25-7.24(m，1H)，7.23(s，1H)，7.22(d，J＝7.1Hz，1H)，7.19(d，J＝7.3Hz，1H)，7.16(t，J＝7.6Hz，1H)，7.09(t，J＝7.5Hz，1H)，6.93(d，J＝2.3Hz，1H)，6.45(d，J＝12.4Hz，1H)，6.15(d，J＝7.9Hz，1H)，5.59(d，J＝12.5Hz，1H)，5.03(dt，J＝8.6，5.3Hz，1H)，3.74(s，3H)，3.37(d，J＝5.2Hz，2H)，3.02(t，J＝2.6Hz，4H). ¹³ C NMR(151MHz，Chloroform-d)δ172.40，165.92，155.02，141.49，140.73，136.07，135.11，130.67，130.31，128.97，128.49(2C)，128.46(2C)，127.57，126.34，126.13，122.65，122.35，119.80，119.40，118.51，118.15，117.11，116.39，111.25，110.01，52.92，52.47，35.59，28.44，27.62.ESI-MS：m/z 553.2[M+H] ⁺ .

(Z) - (3- (1- (3-chlorophenyl) -3-phenethyl-1H-pyrazol-4-yl) acryloyl) -L-tryptophan (I-53)

¹ H NMR(600MHz，DMSO-d ₆ )δ12.70(s，1H)，10.85(d，J＝5.2Hz，1H)，9.25(d，J＝10.1Hz，1H)，8.49(d，J＝7.5Hz，1H)，7.79(s，1H)，7.67(d，J＝8.2Hz，1H)，7.59(d，J＝8.0Hz，1H)，7.51(td，J＝8.2，2.5Hz，1H)，7.35(dt，J＝14.5，7.5Hz，2H)，7.29(d，J＝4.4Hz，4H)，7.18(d，J＝7.8Hz，2H)，7.07(t，J＝7.2Hz，1H)，7.00(q，J＝6.9Hz，1H)，6.61(d，J＝12.8Hz，1H)，5.96(d，J＝12.2Hz，1H)，4.65-4.61(m，1H)，3.30-3.24(m，1H)，3.12-3.06(m，1H)，3.01-2.95(m，4H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ173.51，165.53，154.57，141.15，140.19，135.94，133.86，131.24，130.43，128.26(2C)，128.08(2C)，127.41，127.01，125.85，125.78，123.44，120.78，119.38，118.23，118.02，117.95，116.58(2C)，111.24，109.87，52.93，34.51，27.46，26.98.ESI-MS：m/z 539.2[M+H] ⁺

(E) - (3- (1- (3-chlorophenyl) -3-phenethyl-1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-54)

¹ H NMR(600MHz，Chloroform-d)δ8.12(s，1H)，7.96(s，1H)，7.71(t，J＝1.9Hz，1H)，7.57-7.50(m，2H)，7.47(d，J＝15.6Hz，1H)，7.40-7.33(m，2H)，7.28(s，2H)，7.25(s，1H)，7.23-7.15(m，4H)，7.09(t，J＝7.5Hz，1H)，7.01-6.98(m，1H)，6.07-6.01(m，2H)，5.09(dd，J＝8.2，4.8Hz，1H)，3.73(s，3H)，3.453.34(m，2H)，3.05(s，4H). ¹³ C NMR(151MHz，Chloroform-d)δ172.43，165.48，153.81，141.36，140.47，136.11，135.35，131.11，130.52，128.53(2C)，128.45(2C)，127.76，126.64，126.27，126.14，122.77，122.33，119.79，119.33，119.05，118.71，118.36，116.79，111.28，110.17，53.17，52.43，34.92，29.23，27.74.ESI-MS：m/z 553.2[M+H] ⁺

(E) - (3- (1- (3-chlorophenyl) -3-phenethyl-1H-pyrazol-4-yl) acryloyl) -L-tryptophan (I-55)

¹ H NMR(600MHz，DMSO-d ₆ )δ10.85(s，1H)，8.84(s，1H)，8.40(t，J＝6.1Hz，1H)，7.92(s，1H)，7.81(d，J＝8.2Hz，1H)，7.57(dd，J＝8.3，3.9Hz，1H)，7.53(td，J＝8.2，2.3Hz，1H)，7.39-7.26(m，7H)，7.217.15(m，2H)，7.06(t，J＝7.5Hz，1H)，7.00-6.95(m，1H)，6.47(dd，J＝16.3，4.9Hz，1H)，4.63(t，J＝7.4Hz，1H)，3.25(dd，J＝14.1，6.3Hz，1H)，3.09(q，J＝7.4Hz，1H)，3.03(s，4H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ173.45，165.07，152.51，141.14，140.08，135.94，133.85，131.12，128.88，128.31(3C)，128.11(2C)，127.05，125.78(2C)，123.35，120.76，120.45，118.20，118.05，117.97，117.65，116.43，111.21，109.90，53.05，33.60，28.64，27.07.ESI-MS：m/z 539.2[M+H] ⁺

Example 13

To a 50mL eggplant-shaped bottle was added 28.5mg,0.05mmol of (Z) - (3- (3- (4-nitrophenyl) -1- (m-tolyl) -1H-pyrazol-4-yl) -L-tryptophan methyl ester, methanol (10 mL), iron powder (14 mg,0.25 mmol), an aqueous ammonium chloride solution (8 mg,0.15 mmol), and the mixture was heated under reflux for 5 hours; the iron powder was removed by suction filtration through celite, water (10 mL), dichloromethane (10 mL) was extracted 3 times, dried, filtered, concentrated, and petrol ether: ethyl acetate=1:1 column chromatography to give (Z) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (27 mg, 100%) as a solid. Hydrolysis with methanol gives (Z) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan. The same procedure gave (E) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester and (E) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan

(Z) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-56)

¹ H NMR(600MHz，DMSO-d ₆ )δ10.87(s，1H)，9.18(s，1H)，8.63(d，J＝7.5Hz，1H)，7.86(t，J＝2.0Hz，1H)，7.74(d，J＝8.1Hz，1H)，7.53(dt，J＝7.9，4.1Hz，2H)，7.41-7.30(m，2H)，7.28(d，J＝8.0Hz，2H)，7.19(s，1H)，7.07(t，J＝7.5Hz，1H)，7.00(t，J＝7.4Hz，1H)，6.68(d，J＝8.0Hz，2H)，6.59(d，J＝12.5Hz，1H)，5.98(d，J＝12.6Hz，1H)，5.40(s，2H)，4.66(q，J＝7.4Hz，1H)，3.63(s，3H)，3.26-3.20(m，1H)，3.14-3.07(m，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ172.53，165.61，154.40，149.09，140.24，135.95，133.87，131.24，130.60，129.43(2C)，128.69，126.87，125.97，123.58，120.84，119.68，118.71，118.29，118.05，117.85，116.77，115.58，113.52(2C)，111.31，109.39，53.04，51.76，26.92.ESI-MS：m/z 540.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₀ H ₂₆ ClN ₅ O ₃ [M+H] ⁺ 540.1797，found 540.1796.

(Z) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (I-57)

¹ H NMR(600MHz，DMSO-d ₆ )δ10.85(s，1H)，9.23(s，1H)，8.48(d，J＝7.8Hz，1H)，7.86(s，1H)，7.72(d，J＝8.2Hz，1H)，7.57(d，J＝7.9Hz，1H)，7.53(t，J＝8.1Hz，1H)，7.39(dd，J＝8.0，2.0Hz，1H)，7.33(d，J＝8.1Hz，1H)，7.28(d，J＝8.2Hz，2H)，7.18(s，1H)，7.06(t，J＝7.5Hz，1H)，6.99(t，J＝7.4Hz，1H)，6.68(d，J＝8.2Hz，2H)，6.57(d，J＝12.6Hz，1H)，5.98(d，J＝12.6Hz，1H)，4.61(td，J＝8.4，4.8Hz，1H)，3.25(dd，J＝14.7，4.8Hz，1H)，3.08(dd，J＝14.7，9.1Hz，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ173.46，165.49，154.44，149.04，140.24，135.93，133.87，131.25，130.71，129.44(2C)，128.47，126.99，125.96，123.46，120.77，119.88，118.74，118.23，118.08，118.01，116.73，115.62，113.53(2C)，111.23，109.84，52.92，26.94.ESI-MS：m/z 526.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₂₉ H ₂₄ ClN ₅ O ₃ [M+H] ⁺ 526.1640，found 526.1627.

(E) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-58)

¹ H NMR(600MHz，DMSO-d ₆ )δ10.88(s，1H)，8.98(s，1H)，8.56(d，J＝7.5Hz，1H)，8.03(d，J＝2.5Hz，1H)，7.91(d，J＝8.2Hz，1H)，7.577.51(m，2H)，7.40(d，J＝7.4Hz，2H)，7.37-7.33(m，1H)，7.32-7.28(m，2H)，7.18(s，1H)，7.08(t，J＝7.5Hz，1H)，7.00(t，J＝7.6Hz，1H)，6.69(d，J＝8.0Hz，2H)，6.51(dd，J＝15.7，2.0Hz，1H)，5.42(s，2H)，4.64(q，J＝7.5Hz，1H)，3.62(s，3H)，3.22(dd，J＝14.8，5.4Hz，1H)，3.12(dd，J＝14.6，8.5Hz，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ172.46，165.13，152.92，149.15，140.18，135.96，133.87，131.13，129.99，128.94(2C)，127.41，126.87，125.96，123.52，120.84，120.59，118.86，118.29，117.91，117.84，117.18，116.70，113.62(2C)，111.30，109.46，53.17，51.71，26.86.ESI-MS：m/z 540.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₀ H ₂₆ ClN ₅ O ₃ [M+H] ⁺ 540.1797，found 540.1790.

(E) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (I-59)

¹ H NMR(600MHz，DMSO-d ₆ )δ10.85(s，1H)，8.96(s，1H)，8.40(d，J＝7.8Hz，1H)，8.02(s，1H)，7.90(d，J＝8.2Hz，1H)，7.59-7.53(m，2H)，7.41-7.36(m，2H)，7.36-7.32(m，2H)，7.30(d，J＝8.0Hz，2H)，7.17(s，1H)，7.07(t，J＝7.5Hz，1H)，6.99(t，J＝7.5Hz，1H)，6.69(d，J＝8.0Hz，2H)，6.52(d，J＝15.7Hz，1H)，4.60(td，J＝8.5，4.8Hz，1H)，3.24(dd，J＝14.7，4.9Hz，1H)，3.09(dd，J＝14.6，9.5Hz，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ173.43，165.04，152.89，149.17，140.18，135.94，133.86，131.12，129.67，128.93(2C)，127.31，126.98，125.93，123.41，120.94，120.78，118.85，118.23，118.00，117.90，117.23，116.68，113.59(2C)，111.23，109.90，53.02，26.89.ESI-MS：m/z 526.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₂₉ H ₂₄ ClN ₅ O ₃ [M+H] ⁺ 526.1640，found 526.1632.

Example 14

To a 50mL two-necked flask was added (Z) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (54 mg,0.1 mmol), anhydrous DMF (8 mL), DIPEA (39 mg,0.3 mmol), 1-bromo-2- (2-bromoethoxy) ethane (30 mg,0.13 mmol) under nitrogen and stirring under heating at 75deg.C for 6H; adding water (10 mL), extracting with ethyl acetate (10 mL) for 3 times, drying, filtering, and concentrating; petroleum ether ethyl acetate=1:1 column chromatography gives solid (Z) - (3- (1- (3-chlorophenyl) -3- (4-morpholinophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (16 mg, 27%). Hydrolysis with methanol gives (Z) - (3- (1- (3-chlorophenyl) -3- (4-morpholinophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan. The same procedure gave (E) - (3- (1- (3-chlorophenyl) -3- (4-morpholinophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester and (E) - (3- (1- (3-chlorophenyl) -3- (4-morpholinophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan

(Z) - (3- (1- (3-chlorophenyl) -3- (4-morpholinophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-60)

¹ H NMR(600MHz，CDCl3)δ9.10(s，1H)，8.06(s，1H)，7.80(s，1H)，7.64(d，J＝8.1Hz，1H)，7.57(s，2H)，7.50(d，J＝7.7Hz，1H)，7.33(t，J＝8.0Hz，1H)，7.24(d，J＝8.0Hz，2H)，7.09(dt，J＝29.3，7.2Hz，4H)，6.92(s，1H)，6.66(s，1H)，6.33-6.23(m，1H)，5.71(s，1H)，5.04(d，J＝5.5Hz，1H)，3.94(s，4H)，3.75(s，3H)，3.37(s，2H)，3.27(s，4H). ¹³ C NMR(151MHz，CDCl ₃ )δ171.81，165.35，153.71，140.01，135.44，134.48，130.19，129.68(2C)，129.48(2C)，126.85，125.90(2C)，122.05，121.73(2C)，119.18，118.83(2C)，118.70，117.79，116.54，115.28，110.65(2C)，109.29，65.98(2C)，52.29，51.90，48.96，29.07，26.91.ESI-MS：m/z 610.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₄ H ₃₂ ClN ₅ O ₄ [M+H] ⁺ 610.2216，found 610.2195.

(Z) - (3- (1- (3-chlorophenyl) -3- (4-morpholinophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (I-61)

¹ H NMR(600MHz，DMSO-d ₆ )δ12.74(s，1H)，10.85(s，1H)，9.27-9.24(m，1H)，8.50(d，J＝7.6Hz，1H)，7.88(s，1H)，7.77-7.72(m，1H)，7.58(dd，J＝8.2，2.9Hz，1H)，7.54(t，J＝8.1Hz，1H)，7.48(d，J＝8.4Hz，2H)，7.41(d，J＝7.9Hz，1H)，7.33(d，J＝8.0Hz，1H)，7.19(s，1H)，7.08-7.04(m，3H)，6.99(t，J＝7.5Hz，1H)，6.58(d，J＝12.5Hz，1H)，6.04-5.99(m，1H)，4.62(dd，J＝8.5，5.0Hz，1H)，3.77(t，J＝4.6Hz，3H)，3.26(dd，J＝14.7，4.8Hz，1H)，3.21-3.17(m，4H)，3.08(ddd，J＝14.8，9.5，3.6Hz，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ173.48，165.42，153.67，150.90，140.19，135.93，133.90，131.27，130.88，129.33(2C)，128.13，127.01，126.14，123.46，122.04，120.77，120.35，118.22，118.21，118.02，116.86，115.86，114.56(2C)，111.23，109.87，65.87(2C)，52.97，47.78(2C)，26.96.ESI-MS：m/z 596.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₃ H ₃₀ ClN ₅ O ₄ [M+H] ⁺ 596.2059，found 596.2044.

(E) - (3- (1- (3-chlorophenyl) -3- (4-morpholinophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-62)

¹ H NMR(600MHz，CDCl ₃ )δ8.09(s，1H)，7.79(s，1H)，7.68-7.56(m，4H)，7.56-7.48(m，2H)，7.44-7.35(m，2H)，7.28(s，2H)，7.21-7.15(m，2H)，7.14-7.06(m，2H)，7.05-6.94(m，1H)，6.16(s，1H)，5.10(s，1H)，3.97(s，4H)，3.73(s，3H)，3.40(s，2H)，3.26(s，4H). ¹³ C NMR(151MHz，CDCl ₃ )δ172.51，165.42，140.38，136.18，135.39，130.62(2C)，129.75(2C)，127.75，126.90(2C)，123.07，122.23(3C)，119.69(2C)，119.50(2C)，118.67(2C)，118.09，117.04，111.42(2C)，109.99，72.14，70.92，61.73，53.00，52.54，29.70，27.76.ESI-MS：m/z 610.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₄ H ₃₂ ClN ₅ O ₄ [M+H] ⁺ 610.2216，found 610.2188.

(E) - (3- (1- (3-chlorophenyl) -3- (4-morpholinophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (I-63)

¹ H NMR(600MHz，DMSO-d ₆ )δ10.85(s，1H)，9.00(s，1H)，8.38(d，J＝7.8Hz，1H)，8.04(s，1H)，7.91(d，J＝8.2Hz，1H)，7.59-7.47(m，4H)，7.43-7.36(m，1H)，7.36-7.32(m，2H)，7.17(s，1H)，7.10-7.04(m，3H)，6.98(t，J＝7.5Hz，1H)，6.54(d，J＝15.7Hz，1H)，4.59(dt，J＝8.9，4.8Hz，1H)，3.76(t，J＝4.8Hz，4H)，3.25(dd，J＝14.7，4.5Hz，1H)，3.19(t，J＝4.9Hz，4H)，3.15-3.06(m，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ173.54，164.90，152.15，150.95，140.13，135.93，133.88，131.15，129.27，128.82(2C)，127.51，127.05，126.11，123.38，122.13，121.46，120.74，118.20，118.02，117.52，116.80，114.65(2C)，114.47，111.20，110.02，65.86(2C)，53.20，47.74(2C)，26.93.ESI-MS：m/z 596.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₃ H ₃₀ ClN ₅ O ₄ [M+H] ⁺ 596.2059，found 596.2041.

Example 15

To a 50mL eggplant-shaped bottle was added (Z) - (3- (1- (3-chlorophenyl) -3- (4-methoxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (55 mg,0.1 mmol 1), methylene chloride (5 mL), boron tribromide (5 mL), and the mixture was stirred at room temperature for 5H; methanol quench reaction, dichloromethane: methanol=20:1 column chromatography gave solid (Z) - (3- (1- (3-chlorophenyl) -3- (4-hydroxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (37 mg, 67%). Hydrolysis with methanol gives (Z) - (3- (1- (3-chlorophenyl) -3- (4-hydroxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan. The same procedure gave (E) - (3- (1- (3-chlorophenyl) -3- (4-hydroxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester and (E) - (3- (1- (3-chlorophenyl) -3- (4-hydroxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan.

(Z) - (3- (1- (3-chlorophenyl) -3- (4-hydroxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-64)

¹ H NMR(600MHz，DMSO-d ₆ )δ10.87(s，1H)，9.74(s，1H)，9.21(s，1H)，8.65(d，J＝7.5Hz，1H)，7.89(s，1H)，7.76(d，J＝8.2Hz，1H)，7.57-7.52(m，2H)，7.47-7.38(m，3H)，7.38-7.33(m，1H)，7.19(s，1H)，7.07(t，J＝7.4Hz，1H)，7.00(t，J＝7.4Hz，1H)，6.91(d，J＝8.0Hz，2H)，6.59(d，J＝12.6Hz，1H)，6.01(d，J＝12.5Hz，1H)，4.66(dq，J＝16.0，7.4Hz，1H)，3.64(s，3H)，3.24(dd，J＝14.6，5.4Hz，1H)，3.12(dd，J＝14.4，8.3Hz，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ172.50，165.55，157.76，153.78，140.17，135.95，133.90，131.25，130.73，129.92(2C)，128.34，126.88，126.15，123.58，122.35，120.84，120.11，118.29，118.17，117.85，116.90，115.77，115.33(2C)，111.31，109.38，53.05，51.76，26.92.ESI-MS：m/z 541.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₀ H ₂₅ ClN ₄ O ₄ [M+H] ⁺ 541.1637，found 541.1621.

(Z) - (3- (1- (3-chlorophenyl) -3- (4-hydroxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (I-65)

¹ H NMR(600MHz，DMSO-d ₆ )δ12.68(s，1H)，10.84(s，1H)，9.74(s，1H)，9.26(s，1H)，8.50(d，J＝7.7Hz，1H)，7.88(d，J＝2.2Hz，1H)，7.74(d，J＝8.3Hz，1H)，7.63-7.50(m，2H)，7.47-7.39(m，3H)，7.33(d，J＝8.4Hz，1H)，7.18(s，1H)，7.10-7.04(m，1H)，7.04-6.97(m，1H)，6.90(d，J＝8.0Hz，2H)，6.57(d，J＝12.6Hz，1H)，6.01(d，J＝12.6Hz，1H)，4.62(td，J＝8.8，5.1Hz，1H)，3.26(dd，J＝14.8，5.1Hz，1H)，3.08(dd，J＝14.8，9.0Hz，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ173.45，165.43，157.74，153.83，140.18，135.93，133.90，131.27，130.85，129.94(2C)，129.44，128.13，127.00，126.15，123.46，122.36，120.77，120.31，118.22，118.01，116.86，115.80，115.32(2C)，111.23，109.84，52.93，26.94.ESI-MS：m/z 527.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₂₉ H ₂₃ ClN ₄ O ₄ [M+H] ⁺ 527.1481，found 527.1460.

(E) - (3- (1- (3-chlorophenyl) -3- (4-hydroxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-66)

¹ H NMR(600MHz，DMSO-d ₆ )δ10.99(s，1H)，9.90(s，1H)，9.03(s，1H)，8.63(d，J＝7.5Hz，1H)，8.04(s，1H)，7.92(d，J＝8.2Hz，1H)，7.58-7.50(m，2H)，7.44(d，J＝7.9Hz，2H)，7.40(d，J＝8.0Hz，1H)，7.38-7.33(m，2H)，7.20(s，1H)，7.07(t，J＝7.6Hz，1H)，6.99(t，J＝7.5Hz，1H)，6.94(d，J＝7.9Hz，2H)，6.55(d，J＝15.7Hz，1H)，4.63(q，J＝7.3Hz，1H)，3.62(s，3H)，3.22(dd，J＝14.7，5.2Hz，1H)，3.13(dd，J＝14.6，9.3Hz，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ172.44，165.10，157.90，152.35，140.10，135.96，133.89，131.16，129.62，129.42(2C)，127.64，126.84，126.15，123.60，122.40，120.97，120.80，118.27，118.02，117.79，117.38，116.81，115.48(2C)，111.33，109.38，53.22，51.71，26.83.ESI-MS：m/z 541.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₀ H ₂₅ ClN ₄ O ₄ [M+H] ⁺ 541.1637，found 541.1622.

(E) - (3- (1- (3-chlorophenyl) -3- (4-hydroxyphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (I-67)

¹ H NMR(600MHz，DMSO-d ₆ )δ12.60(s，1H)，10.85(d，J＝2.4Hz，1H)，9.76(s，1H)，9.00(s，1H)，8.42(d，J＝7.9Hz，1H)，8.04(s，1H)，7.92(d，J＝8.3Hz，1H)，7.59-7.51(m，2H)，7.47-7.43(m，2H)，7.41(dd，J＝7.9，1.9Hz，1H)，7.38-7.32(m，2H)，7.17(d，J＝2.5Hz，1H)，7.07(t，J＝7.5Hz，1H)，6.99(t，J＝7.5Hz，1H)，6.94-6.89(m，2H)，6.53(d，J＝15.7Hz，1H)，4.60(tt，J＝8.3，3.8Hz，1H)，3.24(dd，J＝14.7，4.7Hz，1H)，3.09(dd，J＝15.0，9.6Hz，1H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ173.42，164.96，157.81，152.29，140.12，135.95，133.89，131.15，129.44(2C)，129.34，127.55，126.98，126.13，123.41，122.49，121.29，120.78，118.23，118.03，118.00，117.43，116.81，115.44(2C)，111.24，109.90，53.04，26.89.ESI-MS：m/z 527.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₂₉ H ₂₃ ClN ₄ O ₄ [M+H] ⁺ 527.1481，found 527.1464.

Example 16

To a 50mL eggplant flask was added (E) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (I-58, 73mg,0.135 mmol), anhydrous DMF (10 mL), HATU (77 mg,0.202 mmol), DIPEA (67. Mu.L, 0.405 mmol), 3- (2- ((tert-butoxycarbonyl) amino) ethoxy) propionic acid (35 mg,0.16 mmol), and stirred at room temperature overnight. Water (20 mL) was added, extraction was performed 3 times with ethyl acetate (20 mL), drying, filtration, and concentration. Petroleum ether: ethyl acetate=1:1 column chromatography gives solid I-58-Linker-NHBoc. Subsequently, 50mL eggplant-shaped bottle was charged with I-58-Linker-NHBoc, anhydrous dichloromethane (2 mL), trifluoroacetic acid (2 mL), and stirred at room temperature for 1h. Aqueous sodium bicarbonate (5 mL) was added, extracted 3 times with dichloromethane (5 mL), dried, filtered, concentrated and put into the next reaction. Anhydrous DMF (10 mL), HATU (68 mg,0.178 mmol), DIPEA (60. Mu.L, 0.365 mmol), brix An Yisuan (48 mg,0.144 mmol) and stirring overnight at room temperature. Water (10 mL) was added, extraction was performed 3 times with ethyl acetate (10 mL), drying, filtration, and concentration. Column chromatography with dichloromethane: methanol=20:1 gives solid I-58-PROTAC2, which hydrolyzes methyl ester to give I-59-PROTAC2.

I-58-PROTAC2(I-68)

¹ H NMR(600MHz，DMSO-d ₆ )δ11.12(s，1H)，10.87(s，1H)，10.12(s，1H)，9.05(s，1H)，8.59(d，J＝7.6Hz，1H)，8.078.02(m，2H)，7.93(d，J＝8.3Hz，1H)，7.80(t，J＝7.9Hz，1H)，7.74(d，J＝8.2Hz，2H)，7.61-7.50(m，4H)，7.47(d，J＝7.3Hz，1H)，7.43(d，J＝8.1Hz，1H)，7.41-7.32(m，3H)，7.18(s，1H)，7.07(t，J＝7.6Hz，1H)，6.99(t，J＝7.6Hz，1H)，6.53(d，J＝15.7Hz，1H)，5.12(dd，J＝12.9，5.4Hz，1H)，4.78(s，2H)，4.63(q，J＝7.5Hz，1H)，3.74(t，J＝6.4Hz，2H)，3.62(s，3H)，3.49(t，J＝6.0Hz，2H)，3.42-3.37(m，2H)，3.22(dd，J＝14.9，5.3Hz，1H)，3.18(s，1H)，3.11(dd，J＝14.7，8.9Hz，1H)，2.89(ddd，J＝18.1，14.1，5.4Hz，1H)，2.64-2.53(m，4H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ172.60，172.42，169.72，169.21，166.77，166.55，165.27，164.98，154.84，151.76，140.05，139.43，136.73，135.95，133.90，132.86，131.19，129.41，128.50(2C)127.83，126.85，126.31(2C)，123.52，121.24，120.83，120.18，118.95(2C)，118.29，118.14，117.83，117.55，116.92，116.61，115.86，111.29，109.43，68.43，67.35，66.21，64.75，53.18，51.72，48.63，36.91，30.78，26.86，21.83.ESI-MS：m/z 969.2[M+H] ⁺ ，

I-59-PROTAC2(I-69)

¹ H NMR(600MHz，DMSO-d ₆ )δ12.61(s，1H)，10.84(s，1H)，10.14(s，1H)，9.03(s，1H)，8.49-8.41(m，1H)，8.05(s，1H)，7.97-7.86(m，2H)，7.81-7.73(m，2H)，7.60-7.55(m，4H)，7.49-7.40(m，2H)，7.39-7.33(m，2H)，7.29-7.25(m，1H)，7.23-7.14(m，2H)，7.07(t，J＝7.5Hz，1H)，6.99(t，J＝7.5Hz，1H)，6.82(s，1H)，6.54(d，J＝15.7Hz，1H)，4.64-4.49(m，3H)，4.33(q，J＝8.0，7.5Hz，1H)，3.72(t，J＝6.3Hz，2H)，3.47(t，J＝6.0Hz，2H)，3.33-3.30(m，6H)，3.27-3.16(m，1H)，3.09(dd，J＝14.8，9.2Hz，1H)，2.60(q，J＝6.9Hz，2H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ173.55，173.39，173.05，171.85，170.18，169.23，166.26，164.91，154.36，151.76，140.06，139.43，135.94，134.61，133.90，131.18，130.89，129.85，129.62，129.08，128.53(2C)，127.77，126.97，126.42，126.29，123.41，121.61，120.78，120.27，118.98(2C)，118.23，118.13，118.00，117.63，116.92，111.23，109.88，68.41，66.23，59.59，53.03，52.04，36.91，31.33，26.89，20.89，20.60.ESI-MS：m/z 955.2[M+H] ⁺ ，

Example 17

The procedure of example 16 was repeated except for changing (E) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester to (z) - (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester and changing 3- (2- ((tert-butoxycarbonyl) amino) ethoxy) propionic acid to N-Boc-3- [2- (2-aminoethoxy) ethoxy ] propionic acid to obtain the product I-56-PROTAC1 (I-70).

I-56-PROTAC1(I-70)

¹ H NMR(600MHz，DMSO-d ₆ )δ11.12(s，1H)，10.95(s，1H)，10.29(s，1H)，9.20(s，1H)，8.74-8.70(m，1H)，8.10-8.07(m，1H)，7.89(s，1H)，7.83-7.75(m，4H)，7.58-7.50(m，4H)，7.48(d，J＝7.2Hz，1H)，7.44-7.37(m，2H)，7.34(d，J＝8.1Hz，1H)，7.20(s，1H)，7.06(t，J＝7.5Hz，1H)，6.99(t，J＝7.4Hz，1H)，6.60(d，J＝12.5Hz，1H)，6.04(d，J＝12.6Hz，1H)，5.12(dd，J＝12.9，5.4Hz，1H)，4.80(s，2H)，4.64(q，J＝7.5Hz，1H)，3.72(t，J＝6.3Hz，2H)，3.63(s，3H)，3.563.52(m，4H)，3.47(t，J＝5.7Hz，2H)，3.31(q，J＝5.8Hz，2H)，3.25-3.15(m，2H)，3.11(dd，J＝14.6，8.8Hz，1H)，2.90(ddd，J＝18.0，13.9，5.5Hz，1H)，2.63-2.57(m，4H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ172.62，172.49，169.72，169.34，166.75，166.57，165.50，165.27，154.84，153.21，140.11，139.46，136.76，135.94，133.89，132.86，131.29，130.88，128.92(2C)，128.09，126.84，126.29，126.20，123.64，120.79，120.48，120.15，118.82(2C)，118.26(2C)，117.80，117.02，116.55，115.95，115.83，111.32，109.32，69.44，69.39，68.65，67.29，66.47，53.09，51.76，48.63，38.23，37.01，30.78，26.87，21.83.ESI-MS：m/z 1013.2[M+H] ⁺ ，

Example 18

The (E) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester was converted to (Z) - (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester, and the remaining required raw materials, reagents and preparation method were the same as in example 16 to give product I-58-PROTAC1 (I-71), and hydrolysis gave I-59-PROTAC1 (I-72).

I-58-PROTAC1(I-71)

¹ H NMR(600MHz，DMSO-d ₆ )δ11.12(s，1H)，10.87(s，1H)，10.13(s，1H)，9.05(s，1H)，8.58(d，J＝7.6Hz，1H)，8.05(s，1H)，8.027.99(m，1H)，7.93(d，J＝8.3Hz，1H)，7.79(t，J＝8.0Hz，1H)，7.75(d，J＝8.3Hz，2H)，7.597.55(m，3H)，7.52(d，J＝8.0Hz，1H)，7.48(d，J＝7.1Hz，1H)，7.43(d，J＝8.1Hz，1H)，7.41-7.33(m，3H)，7.18(s，1H)，7.07(t，J＝7.5Hz，1H)，6.99(t，J＝7.4Hz，1H)，6.53(d，J＝15.4Hz，1H)，5.12(dd，J＝12.9，5.4Hz，1H)，4.79(s，2H)，4.63(q，J＝7.4Hz，1H)，3.72(t，J＝6.3Hz，2H)，3.62(s，3H)，3.54(d，J＝5.3Hz，4H)，3.47(t，J＝5.7Hz，2H)，3.31(q，J＝5.8Hz，2H)，3.25-3.16(m，2H)，3.11(dd，J＝14.8，8.8Hz，1H)，2.89(ddd，J＝18.2，13.8，5.4Hz，1H)，2.62-2.57(m，4H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ172.61，172.42，169.71，169.29，166.73，166.56，165.27，164.99，154.81，151.78，140.05，139.45，136.74，135.95，133.91，132.86，131.19，129.40，128.53(2C)，127.85，126.86，126.37，126.31，123.52，121.24，120.84，120.15，118.94(2C)，118.29，118.14，117.83，117.57，116.92，116.58，115.85，111.29，109.43，69.45，69.39，68.66，67.31，66.44，53.18，51.72，48.63，38.23，37.05，30.78，26.86，21.86.ESI-MS：m/z 1013.2[M+H] ⁺ ，

I-59-PROTAC1(I-72)

¹ H NMR(600MHz，DMSO-d ₆ )δ10.85(s，1H)，10.15(s，1H)，9.04(s，1H)，8.74(d，J＝8.0Hz，1H)，8.43(d，J＝8.0Hz，1H)，8.05(s，1H)，7.94(d，J＝8.2Hz，1H)，7.77(d，J＝8.2Hz，2H)，7.62-7.55(m，4H)，7.48-7.37(m，3H)，7.37-7.32(m，2H)，7.28(d，J＝8.6Hz，1H)，7.21(d，J＝8.7Hz，1H)，7.19-7.16(m，1H)，7.07(t，J＝7.6Hz，1H)，6.99(t，J＝7.5Hz，1H)，6.82(d，J＝9.4Hz，1H)，6.54(d，J＝15.4Hz 1H)，4.64-4.50(m，3H)，4.33(t，J＝8.7Hz，1H)，3.72(q，J＝5.0，3.9Hz，2H)，3.54-3.50(m，5H)，3.48-3.43(m，3H)，3.42-3.35(m，2H)，3.30(t，J＝5.6Hz，2H)，3.24(dd，J＝14.6，4.7Hz，1H)，3.15-3.05(m，1H)，2.60(t，J＝6.4Hz，2H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ173.56，173.40，173.06，170.18，169.29，167.58，167.43，164.91，154.35，151.76，140.06，139.45，135.94，134.61，133.91，131.18，129.83，129.08，128.53(2C)，127.78，126.98，126.41，126.29，124.55，123.41，121.62，120.78，120.25，118.95(2C)，118.23，118.13，118.00，117.63，116.92，115.56，111.23，109.89，69.40，69.36，68.61，67.73，66.44，53.03，52.03，38.22，37.04，31.33，26.89，20.89.ESI-MS：m/z 999.2[M+H] ⁺ ，

Example 19

The (E) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester was converted to (Z) - (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester, and the remaining required raw materials, reagents and preparation method were the same as in example 16 to give a product I-56-PROTAC2 (I-73), and hydrolysis gave I-57-PROTAC2 (I-74).

I-56-PROTAC2(I-73)

¹³ C NMR(151MHz，Chloroform-d)δ171.93，170.46，169.60，168.04，167.96，166.18，165.62，165.42，153.47，152.99，139.85，137.81，136.51，135.49，134.49，132.72，130.08，129.73，128.87(2C)，128.72，127.07，126.63，125.99，122.21，121.58，119.58，119.08(2C)，119.06，118.73，117.80，117.11，116.68，116.50，115.30，115.24，110.65，108.95，69.15，66.90，66.29，65.24，51.92，48.64，38.16，37.59，30.64，26.83，21.86.ESI-MS：m/z 969.2[M+H] ⁺ ，

I-57-PROTAC2(I-74)

¹ H NMR(600MHz，DMSO-d ₆ )δ12.70(s，1H)，10.85(s，1H)，10.12(s，1H)，9.27(s，1H)，8.53(d，J＝7.8Hz，1H)，7.89(s，1H)，7.78-7.73(m，3H)，7.59-7.50(m，4H)，7.49-7.44(m，1H)，7.42(d，J＝8.1Hz，1H)，7.33(d，J＝8.3Hz，1H)，7.30-7.20(m，2H)，7.18(s，1H)，7.06(t，J＝7.6Hz，1H)，6.99(t，J＝7.5Hz，1H)，6.83(s，1H)，6.59(d，J＝12.5Hz，1H)，6.03(d，J＝12.6Hz，1H)，4.64-4.49(m，3H)，4.37-4.28(m，1H)，3.72(t，J＝6.4Hz，2H)，3.48(t，J＝6.0Hz，2H)，3.42-3.37(m，1H)，3.33-3.30(m，4H)，3.25(dd，J＝14.9，4.8Hz，1H)，3.18(s，1H)，3.08(dd，J＝14.8，9.2Hz，1H)，2.60(q，J＝7.1Hz，2H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ173.55，173.44，173.06，170.18，169.25，166.90，166.27，165.38，154.36，153.27，140.13，139.35，135.93，134.62，133.91，131.30，130.95，129.86，129.62，129.01(2C)，127.91，126.98，126.30(2C)，123.47，122.17，120.77，120.64，120.27，118.86(2C)，118.31,118.23，118.00，116.98，115.99，111.23，109.82，68.38，67.76，66.25，64.75，52.94，52.16，36.92，31.23，26.89，20.60.ESI-MS：m/z 955.2[M+H] ⁺ ，

Example 20

The (N-Boc-3- [2- (2-aminoethoxy) ethoxy ] propionic acid was changed to 2, 2-dimethyl-4-oxo-3, 8, 11, 14-tetraoxa-5-aza-heptadecane-17-acid, and the remaining required raw materials, reagents and preparation method were the same as in example 17 to give the product I-56-PROTAC3 (I-75), and hydrolysis gave I-57-PROTAC3 (I-76).

I-56-PROTAC3(I-75)

¹ H NMR(600MHz，DMSO-d ₆ )δ11.12(s，1H)，10.87(s，1H)，10.12(s，1H)，9.21(s，1H)，8.66(d，J＝7.5Hz，1H)，8.02-7.99(m，1H)，7.96(s，1H)，7.90(s，1H)，7.82-7.78(m，1H)，7.77-7.73(m，2H)，7.58-7.52(m，4H)，7.49(d，J＝7.3Hz，1H)，7.40(dd，J＝20.4，8.3Hz，2H)，7.33(d，J＝8.1Hz，1H)，7.19(s，1H)，7.07(t，J＝7.6Hz，1H)，7.00(t，J＝7.5Hz，1H)，6.61(d，J＝12.5Hz，1H)，6.02(d，J＝12.6Hz，1H)，5.12(dd，J＝12.9，5.6Hz，1H)，4.78(s，2H)，4.66(q，J＝7.5Hz，1H)，3.72(t，J＝6.3Hz，2H)，3.63(s，3H)，3.54-3.49(m，8H)，3.46(t，J＝5.8Hz，2H)，3.42-3.37(m，1H)，3.33-3.29(m，1H)，3.23(dd，J＝14.7，5.4Hz，1H)，3.10(dd，J＝14.7，8.8Hz，1H)，2.90(s，2H)，2.64-2.53(m，4H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ172.60，172.49，169.70，169.28，166.72，166.56，165.48，165.27，162.14，154.80,153.20，140.11，139.38，136.76,135.94，133.90，132.86，131.29，130.88，128.98(2C)，128.12，126.86，126.30，126.26，123.58，120.83，120.44，120.15，118.81(2C)，118.27，117.84，117.01，116.59，115.94，115.87，111.30，109.36，69.56，69.53，69.45，68.64，67.32，66.44，64.75，53.05，51.77，48.64，37.05，35.62，30.69，26.91，21.83.ESI-MS：m/z 1057.2[M+H] ⁺ ，

I-57-PROTAC3(I-76)

¹ H NMR(600MHz，DMSO-d ₆ )δ12.64(s，1H)，10.84(d，J＝2.5Hz，1H)，10.13(s，1H)，9.27(s，1H)，8.52(d，J＝7.8Hz，1H)，7.89(d，J＝2.3Hz，1H)，7.84(t，J＝5.7Hz，1H)，7.75(d，J＝8.2Hz，3H)，7.56(q，J＝8.1Hz，4H)，7.487.40(m，2H)，7.377.26(m，2H)，7.227.16(m，2H)，7.06(t，J＝7.5Hz，1H)，6.99(t，J＝7.4Hz，1H)，6.82(d，J＝11.6Hz，1H)，6.59(d，J＝12.5Hz，1H)，6.03(d，J＝12.6Hz，1H)，4.64-4.55(m，3H)，4.32(q，J＝6.2，5.4Hz，1H)，3.72(t，J＝6.2Hz，2H)，3.51(d，J＝19.9Hz，9H)，3.44(t，J＝5.9Hz，2H)，3.42-3.37(m，1H)，3.30(t，J＝5.5Hz，1H)，3.28-3.23(m，1H)，3.18(s，1H)，3.08(dd，J＝14.7，9.1Hz，1H)，2.89(s，1H)，2.60(t，J＝6.3Hz，2H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ173.56，173.44，171.85，170.18,169.29，167.44，166.22，165.38，154.35，153.27，140.13，139.38，135.93,134.61，133.91,131.30，131.00，129.84，129.60,129.02(2C)，127.90,126.98，126.29,124.56，123.47，120.77,120.64，120.25，118.82(2C)，118.31，118.23，118.00，116.98，115.98，115.56，111.23，109.82，69.51，69.42，69.37，68.54，67.74，66.45，64.75，52.93，52.03，37.06，31.33，28.70，26.93，20.89.ESI-MS：m/z 1043.2[M+H] ⁺ ，

Example 21

The (Z) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester was converted to (E) - (3- (3- (4-aminophenyl) -1- (3-chlorophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester, and the remaining desired starting materials, reagents and preparation method were the same as in example 20 to give product I-56-PROTAC3 (I-77), and hydrolysis gave I-57-PROTAC3 (I-78).

I-58-PROTAC3(I-77)

¹ H NMR(600MHz，DMSO-d ₆ )δ11.12(s，1H)，10.87(s，1H)，10.14(s，1H)，9.05(s，1H)，8.59(d，J＝7.6Hz，1H)，8.05(s，1H)，8.00(t，J＝5.8Hz，1H)，7.93(d，J＝8.3Hz，1H)，7.82-7.74(m，3H)，7.60-7.55(m，3H)，7.53(d，J＝8.0Hz，1H)，7.49(d，J＝7.3Hz，1H)，7.43(d，J＝8.1Hz，1H)，7.40-7.33(m，3H)，7.18(s，1H)，7.07(t，J＝7.6Hz，1H)，6.99(t，J＝7.5Hz，1H)，6.53(d，J＝15.7Hz，1H)，5.12(dd，J＝12.9，5.4Hz，1H)，4.78(s，2H)，4.64(q，J＝7.5Hz，1H)，3.72(t，J＝6.3Hz，2H)，3.62(s，3H)，3.51(d，J＝10.4Hz，9H)，3.45(t，J＝5.8Hz，2H)，3.42-3.37(m，1H)，3.333.29(m，1H)，3.22(dd，J＝14.8，5.3Hz，1H)，3.12(dt，J＝14.7，8.8Hz，1H)，2.90(td，J＝16.3，14.1，5.5Hz，1H)，2.66-2.52(m，4H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ172.60,172.42,169.70，169.28，166.71,166.56，165.26,164.98，154.81，151.79,140.04，139.47,136.74，135.95，133.90，132.86，131.18，129.40，128.53(2C)，127.85，126.86，126.37，126.30，123.52，121.25，120.83，120.14，118.93(2C)，118.29，118.13，117.83，117.56，116.92，116.58，115.86，111.29，109.43，69.55，69.53，69.44，68.64，67.31，66.43，64.75，53.43，53.18，51.71，48.64，37.04，30.78，26.86，21.83.ESI-MS：m/z 1057.2[M+H] ⁺ ，

I-59-PROTAC3(I-78)

¹ H NMR(600MHz，DMSO-d ₆ )δ12.63(s，1H)，10.84(d，J＝2.3Hz，1H)，10.15(s，1H)，9.03(s，1H)，8.43(d，J＝8.0Hz，1H)，8.05(d，J＝2.2Hz，1H)，7.967.86(m，1H)，7.84(t，J＝5.8Hz，1H)，7.77(d，J＝8.2Hz，2H)，7.57(td，J＝6.2，4.7，2.3Hz，4H)，7.487.40(m，2H)，7.397.29(m，2H)，7.297.19(m，2H)，7.18(d，J＝7.4Hz，1H)，7.07(t，J＝7.5Hz，1H)，6.99(t，J＝7.5Hz，1H)，6.82(d，J＝11.6Hz，1H)，6.54(d，J＝15.7Hz，1H)，4.654.55(m，3H)，4.32(q，J＝6.7，5.8Hz，1H)，3.72(t，J＝6.2Hz，2H)，3.543.47(m，8H)，3.463.42(m，2H)，3.373.34(m，5H)，3.323.27(m，1H)，3.273.20(m，1H)，3.153.04(m，1H)，2.60(t，J＝6.4Hz，2H). ¹³ C NMR(151MHz，DMSO-d ₆ )δ173.56，173.40，173.05，169.29，167.44，166.22，165.69，164.911，154.35，151.76，140.06，139.46，135.94，134.61，133.90，131.18，130.89，129.83，129.59，129.08，128.53(2C)，127.78，126.98，126.41，126.29，124.56，123.41，121.61，120.78，118.95(2C)，118.23，118.13，118.00，117.63，116.92，111.23，109.88，69.51，69.41，69.36，68.54，67.73，67.56，66.43，53.03，52.03，48.43，37.04，31.33，26.89，20.89.ESI-MS：m/z 1042.2[M+H] ⁺ ，

Comparative example 1

(E) - (3- (3- (naphthalen-2-yl) -1-phenyl-1H-pyrazol-4-yl) acryloyl) -L-tryptophan

(comparative Compound 1)

The required raw materials, reagents and preparation method are the same as CN 110746355Example 64 in A gave the comparative compound 1 (E) - (3- (3- (naphthalen-2-yl) -1-phenyl-1H-pyrazol-4-yl) acryloyl) -L-tryptophan. ¹ H NMR(400MHz，Methanol-d ₄ )δ8.68(s，1H)，8.14(d，J＝1.6Hz，1H)，8.03-7.86(m，6H)，7.83(dd，J＝8.5，1.7Hz，1H)，7.68(d，J＝15.7Hz，1H)，7.60(d，J＝7.9Hz，1H)，7.58-7.53(m，4H)，7.40(t，J＝7.4Hz，1H)，7.33(d，J＝8.1Hz，1H)，7.12(s，1H)，7.09(t，J＝7.5Hz，1H)，7.04-6.95(m，1H)，6.55(d，J＝15.7Hz，1H)，4.87-4.83(m，4H)，3.44(dd，J＝14.8，4.5Hz，1H)，3.26(dd，J＝14.7，7.3Hz，1H).MS(ESI，m/z)：525.0[M-H] ^- .

Comparative example 2

(Z) - (3- (3- (naphthalen-2-yl) -1-phenyl-1H-pyrazol-4-yl) acryloyl) -L-tryptophan

(comparative Compound 2)

The desired starting materials, reagents and preparation method CN 110746355A, example 64, gave the comparative compound 2 (Z) - (3- (3- (naphthalen-2-yl) -1-phenyl-1H-pyrazol-4-yl) acryloyl) -L-tryptophan. ¹ H NMR(400MHz，DMSO-d ₆ )δ10.70(s，1H)，9.44(s，1H)，8.12(s，1H)，8.04(d，J＝8.1Hz，2H)，8.007.97(m，1H)，7.88(d，J＝8.1Hz，2H)，7.79(dd，J＝8.5，1.8Hz，2H)，7.59-7.52(m，6H)，7.37(t，J＝7.4Hz，1H)，7.31-7.23(m，1H)，7.10(s，1H)，6.99(dd，J＝8.1，6.9Hz，1H)，6.88(t，J＝7.4Hz，1H)，6.57(d，J＝12.5Hz，1H)，6.00(d，J＝12.6Hz，1H)，4.34-4.27(m，1H)，3.16(d，J＝4.7Hz，1H)，3.02(dd，J＝14.7，7.4Hz，1H).MS(ESI，m/z)：527.2[M+H] ⁺ .

Comparative example 3

(Z) - (3- (3- (2-naphthyl) -1- (m-tolyl) -1H-4-pyrazolyl) acryloyl) -L-tryptophan (comparative compound 3)

Comparative Compound 3 the same Compound (I-1) as in example 6

Comparative example 4

The comparative compound 4 (Z) - (3- (3- (2-naphthyl) -1- (o-tolyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan was obtained by replacing m-tolylhydrazine hydrochloride with o-tolylhydrazine hydrochloride, and the remaining required raw materials, reagents, and preparation methods were the same as in example 6.

(Z) - (3- (3- (2-naphthyl) -1- (o-tolyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (comparative compound 4)

¹ H NMR(600MHz，DMSO-d6)δ12.49(s，1H)，10.84(d，J＝2.4Hz，1H)，8.98(d，J＝0.5Hz，1H)，8.49(d，J＝7.8Hz，1H)，8.15-8.09(m，1H)，8.05-8.01(m，2H)，8.00-7.95(m，1H)，7.76(dd，J＝8.5，1.7Hz，1H)，7.60-7.53(m，3H)，7.48-7.36(m，4H)，7.32(dt，J＝8.1，0.9Hz，1H)，7.17(d，J＝2.4Hz，1H)，7.05(ddd，J＝8.1，6.9，1.2Hz，1H)，6.97(ddd，J＝8.0，7.0，1.1Hz，1H)，6.76-6.71(m，1H)，6.01(d，J＝12.7Hz，1H)，4.58(ddd，J＝9.2，7.8，4.9Hz，1H)，3.24(dd，J＝14.7，4.5Hz，1H)，3.06(dd，J＝14.7，9.1Hz，1H)，2.30(s，3H)； ¹³ C NMR(151MHz，DMSO-d6)δ174.05，172.49，166.19，153.03，139.62，136.56，135.67，133.31，133.22，132.98，131.93，130.27，129.08，129.02，128.73，128.63，128.31，128.07，127.61，127.36，127.15，126.98，126.09，124.07，121.40，120.29，118.85，118.62，115.31，111.86，110.48，53.51，27.55，18.45；ESI-MS：m/z541.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₄ H ₂₉ O ₃ N ₃ [M+H] ⁺ 541.2234，found 541.2234.

Comparative example 5

The comparative compound 5 (Z) - (3- (3- (2-naphthyl) -1- (p-tolyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan was obtained by converting m-tolylhydrazine hydrochloride to p-tolylhydrazine hydrochloride, and the remaining required raw materials, reagents, and preparation methods were the same as in example 6.

(Z) - (3- (3- (2-naphthyl) -1- (p-tolyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (comparative compound 5)

¹ H NMR(600MHz，DMSO-d6)δ12.68(s，1H)，10.86(d，J＝2.4Hz，1H)，9.29(s，1H)，8.53(d，J＝7.8Hz，1H)，8.12(d，J＝1.7Hz，1H)，8.09-8.02(m，2H)，7.99(dt，J＝7.1，3.6Hz，1H)，7.79(dd，J＝8.4，1.7Hz，1H)，7.74-7.70(m，2H)，7.63-7.56(m，3H)，7.34(dd，J＝8.2，6.2Hz，3H)，7.19(d，J＝2.2Hz，1H)，7.07(t，J＝7.3Hz，1H)，7.00(t，J＝7.4Hz，1H)，6.70(d，J＝12.6Hz，1H)，6.03(d，J＝12.6Hz，1H)，4.66-4.61(m，1H)，3.27(dd，J＝14.7，4.8Hz，1H)，3.09(dd，J＝14.7，9.2Hz，1H)，2.37(s，3H)； ¹³ C NMR(151MHz，DMSO-d6)δ174.09，166.15，153.49，137.45，136.74，136.58，133.29，133.05，131.38，130.58(2C)，130.15，128.76，128.65，128.29，128.09，127.63，127.60，127.06(2C)，127.02，124.10，121.42，120.83，119.20(2C)，118.88，118.65，116.44，111.89，110.50，53.57，27.57，20.98；ESI-MS：m/z 541.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₄ H ₂₉ O ₃ N ₄ [M+H] ⁺ 541.2234，found 541.2233.

Comparative example 6 exchange of 2-Naphtholone for p-nitroacetophenone, exchange of m-methyl phenylhydrazine hydrochloride for m-chlorophenylhydrazine hydrochloride, and the remaining desired starting materials, reagents and preparation method are the same as those of example 6, giving comparative compound 6 (Z) - (3- (1- (3-chlorophenyl) -3- (4-nitrophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan and comparative compound 7 (E) - (3- (1- (3-chlorophenyl) -3- (4-nitrophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan

(Z) - (3- (1- (3-chlorophenyl) -3-mono (4-nitrophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (comparative compound 6)

¹ H NMR(600MHz，DMSO-d6)δ12.52(s，1H)，10.84(s，1H)，9.23(s，1H)，8.57(d，J＝7.8Hz，1H)，8.38-8.30(m，2H)，7.98-7.88(m，3H)，7.79(ddd，J＝8.2，2.2，0.9Hz，1H)，7.56(t，J＝8.0Hz，2H)，7.46(ddd，J＝8.1，2.0，0.9Hz，1H)，7.32(dd，J＝8.0，1.0Hz，1H)，7.17(d，J＝2.2Hz，1H)，7.05(ddd，J＝8.1，7.0，1.2Hz，1H)，6.98(ddd，J＝7.9，6.9，1.0Hz，1H)，6.65(d，J＝12.5Hz，1H)，6.11(d，J＝12.5Hz，1H)，4.63-4.56(m，1H)，3.24(dd，J＝14.7，4.8Hz，1H)，3.07(dd，J＝14.7，9.1Hz，1H)； ¹³ C NMR(151 MHz，DMSO-d6)δ173.96，165.63，151.46，147.64，140.53，138.89，136.56，134.61，132.10，131.98，130.06(2C)，127.81，127.61，127.42，124.38(2C)，124.11，122.82，121.41，119.23，118.86，118.64，117.90，117.35，111.86，110.39，53.56，27.58；ESI-MS：m/z 556.1[M+H]+，ESI-HRMS：calcd for C ₂₉ H ₂₃ O ₅ N ₄ Cl[M+H] ⁺ 556.1382，found 556.1381.

(E) - (3- (1- (3-chlorophenyl) -3- (4-nitrophenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (comparative compound 7)

¹ H NMR(600MHz，DMSO-d6)δ12.71(s，1H)，10.86(s，1H)，9.12(s，1H)，8.48(d，J＝7.8Hz，1H)，8.38(d，J＝8.4Hz，2H)，8.08(s，1H)，7.94(dd，J＝14.1，8.2Hz，3H)，7.62-7.54(m，2H)，7.46(d，J＝8.0Hz，1H)，7.38(d，J＝15.6Hz，1H)，7.34(d，J＝8.1Hz，1H)，7.17(s，1H)，7.07(t，J＝7.5Hz，1H)，6.98(t，J＝7.5Hz，1H)，6.57(d，J＝15.7Hz，1H)，4.62(dt，J＝12.5，6.2Hz，1H)，3.25(dd，J＝14.8，4.8Hz，1H)，3.09(dd，J＝14.7，9.3Hz，1H)； ¹³ C NMR(151MHz，DMSO-d6)δ173.98，165.29，150.22，147.77，140.46，138.91，136.58，134.61，131.90，129.75(2C)，129.31，128.88，127.61，127.46，124.58(2C)，124.07，123.46，121.42，119.07，119.02，118.87，118.63，117.83，111.87，110.47，53.69，27.54；ESI-MS：m/z 556.1[M+H] ⁺ ，ESI-HRMS：calcd for C ₂₉ H ₂₃ O ₅ N ₄ Cl[M+H] ⁺ 556.1382，found 556.1384.

Comparative example 7

The esterification product (Z) - (3- (1- (3- (m-chlorophenyl) -3- (4-ethylphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester was retained, and further hydrolyzed to give the comparative compound 8 (Z) - (3- (1- (3- (m-chlorophenyl) -3- (4-ethylphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan by changing 2-naphthaceneone to p-ethylacetophenone and m-chlorophenylhydrazine hydrochloride, and the other desired raw materials, reagents and preparation method were the same as in example 6.

(Z) - (3- (1- (3- (m-chlorophenyl) -3- (4-ethylphenyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan methyl ester (comparative compound 8)

¹ H NMR (600 mhz, dmso-d 6) δ10.87 (d, j=2.4 hz, 1H), 9.22 (s, 1H), 8.67 (d, j=7.5 hz, 1H), 7.89 (t, j=2.1 hz, 1H), 7.77 (ddd, j=8.2, 2.2,0.9hz, 1H), 7.57-7.51 (m, 4H), 7.42 (ddd, j=8.0, 2.1,0.9hz, 1H), 7.38-7.31 (m, 3H), 7.18 (d, j=2.3 hz, 1H), 7.06 (ddd, j=8.1, 6.9,1.2hz, 1H), 6.99 (ddd, j=7.9, 6.9,1.0 hz), 6.60 (d, j=12.5 hz, 1H), 6.02 (ddd, 12.12 hz), 7.38-7.31 (m, 3H), 7.18 (d, j=2.3 hz, 1H), 7.06 (d, 1H), 7.8.9, 6.9, 1H), 6.60 (d, 1H), 6.9 (d, 1H), 6.12.9 (d, 6.9 hz), 6.6.6.6H), 6.6 (1H), 6.7 (1H), 6.7.7.8 (1H), 6.9 (1H), 6.7.7.7.9 (1H), 1H (j=7.6.9, 1H). ¹³ C NMR(151MHz，DMSO-d6)δ173.12，166.12，154.18，144.77，140.76，136.57，134.54，131.93，131.53，129.70，129.20(2C)，128.74，128.56(2C)，127.49，126.97，124.22，121.47，121.07，118.92(2C)，118.47，117.67，116.63，111.94，110.00，53.68，52.40，28.45，27.54，16.00；ESI-MS：m/z 553.2[M+H] ⁺ ，ESI-HRMS：calcd for C ₃₂ H ₃₀ O ₃ N ₄ Cl[M+H] ⁺ 553.2001，found 553.2013.

Comparative example 8

I-8-PROTAC (comparative Compound 9)

To a 50mL eggplant flask was added (E) - (3- (1- (3-chlorophenyl) -3- (2-naphthyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan (I-8, 184mg,0.33 mmol), anhydrous DMF (10 mL), HATU (194 mg,0.5 mmol), DIPEA (112. Mu.L, 0.66 mmol), tert-butyl 2- (2- (2-aminoethoxy) ethoxy) ethylcarbamate (127 mg,0.5 mmol), and stirred at room temperature for 6H. Water (60 mL) was added, and the mixture was extracted 3 times with dichloromethane (20 mL), dried, filtered, and concentrated. Dichloromethane: methanol=20:1 column chromatography gave I-8-Linker-NHBoc (260 mg, 99%) as a yellow oil. ¹ H NMR(400MHz，Chloroform-d)δ8.68(s，1H)，8.27(s，1H)，8.11(s，1H)，8.00(d，J＝3.7Hz，2H)，7.95-7.84(m，3H)，7.79(d，J＝8.2Hz，1H)，7.69(dd，J＝18.3，11.1Hz，2H)，7.51(q，J＝4.3Hz，2H)，7.43-7.31(m，2H)，7.29(d，J＝7.7Hz，1H)，7.19-7.06(m，2H)，6.62(s，1H)，6.35(d，J＝15.5Hz，1H)，6.15(s，1H)，5.95(s，1H)，5.05(s，1H)，4.93-4.74(m，1H)，3.68-3.01(m，7H)，3.04-2.91(m，5H)，2.88-2.85(m，2H)，1.48-1.33(m，9H).

50mL eggplant-shaped flask was charged with I-8-Linker-NHBoc, dichloromethane (4 mL), trifluoroacetic acid (1 mL), and stirred at room temperature for 1h. Aqueous sodium hydroxide was adjusted to pH, extracted 3 times with dichloromethane (5 mL), dried, filtered, concentrated, taken to the next step (189 mg,0.27 mmol), DMF (10 mL), DIPEA (90. Mu.L, 0.54 mmol), F-thalidomide (68.7 mg,0.25 mmol), stirred and heated to reflux for 2h. Water (30 mL) was added, and the mixture was extracted 3 times with dichloromethane (10 mL), dried, filtered, and concentrated. Dichloromethane: methanol=20:1 column chromatography gave I-8-procac (50 mg, 20%) as a yellow oil.

¹ H NMR(600MHz，DMSO-d6)δ11.10(s，1H)，10.81(s，1H)，9.07(s，1H)，8.32(d，J＝8.2Hz，1H)，8.16(s，1H)，8.09(s，2H)，8.04(dd，J＝13.8，6.9Hz，2H)，7.99-7.94(m，2H)，7.78(d，J＝8.5Hz，1H)，7.64(d，J＝7.9Hz，1H)，7.58(d，J＝7.6Hz，3H)，7.52(t，J＝7.8Hz，1H)，7.47-7.42(m，1H)，7.32(d，J＝8.1Hz，1H)，7.17-7.14(m，1H)，7.05(t，J＝7.9Hz，2H)，7.00(d，J＝6.9Hz，1H)，6.96(t，J＝7.4Hz，1H)，6.58(d，J＝16.3Hz，2H)，5.76(s，1H)，5.05(dd，J＝12.9，5.5Hz，1H)，4.65(q，J＝7.9Hz，1H)，3.60-3.40(m，10H)，3.26-3.17(m，2H)，3.15-3.10(m，1H)，3.03-2.96(m，1H)，2.91-2.84(m，2H)，2.73(s，2H)，2.61-2.56(m，1H)； ¹³ C NMR(151MHz，DMSO-d6)δ172.64，171.59，169.92，168.75，167.11，164.66，162.16，151.92，146.17，140.04，135.99，135.89，132.70，132.54，131.88，131.20，129.33，128.76，128.20，128.10，127.93，127.50，127.18，127.13，126.54，126.48，126.42，125.91，123.45，122.19，120.67，118.37，118.23，118.08，118.02，117.16，117.00，111.09，110.47，110.05，109.04，69.46(2C)，68.75，68.69，54.74，53.51，48.38，41.49，30.81，27.86，21.96；ESI-MS：m/z 969.2[M+Na] ⁺ .

Comparative example 9

The (E) - (3- (1- (3-chlorophenyl) -3- (2-naphthyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan was replaced by (Z) - (3- (1- (3-chlorophenyl) -3- (5, 6,7, 8-tetrahydronaphthalen-2-yl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan, and the remaining desired starting materials, reagents and preparation method were the same as in comparative example 8 to give the product I-25-PROTAC.

I-25-PROTAC (comparative Compound 10)

¹ H NMR(600MHz，DMSO-d6)δ11.09(s，1H)，10.82-10.79(m，1H)，9.23(s，1H)，8.43(d，J＝8.0Hz，1H)，8.16(t，J＝5.7Hz，1H)，7.96(s，1H)，7.88(s，1H)，7.77-7.72(m，1H)，7.66(d，J＝7.9Hz，1H)，7.53(t，J＝7.6Hz，2H)，7.39(d，J＝8.1Hz，1H)，7.30(d，J＝8.1Hz，1H)，7.27(d，J＝6.4Hz，2H)，7.17(d，J＝8.6Hz，2H)，7.106.94(m，3H)，6.58-6.50(m，2H)，6.00(d，J＝12.6Hz，1H)，5.05(dd，J＝12.8，5.5Hz，1H)，4.65(td，J＝8.5，5.1Hz，1H)，3.59-3.45(m，5H)，3.43-3.37(m，3H)，3.28-3.21(m，2H)，3.14(dd，J＝14.6，5.2Hz，1H)，2.98(dd，J＝14.5，9.2Hz，1H)，2.89(s，3H)，2.81-2.74(m，4H)，2.73(s，3H)，1.77(dd，J＝7.2，3.6Hz，4H)； ¹³ C NMR(151 MHz，DMSO-d6)δ172.62，171.71，169.90，168.74，167.11，165.33，162.14，153.58，146.17，140.14，137.03，136.81，135.98，135.87，133.90，131.88，131.21，130.73，128.99,128.93，127.23，127.11，126.19，125.70，123.53,121.11，120.66,118.40，118.12，118.00，117.16，116.88，116.09，111.08，110.46,110.02,109.05,69.48,69.45,68.76,68.70,53.43,48.38,41.48,35.61,30.81,30.60,28.60，28.47，27.86，22.49，21.97；ESI-MS：m/z 973.3[M+Na] ⁺ .

Comparative example 10

The (E) - (3- (1- (3-chlorophenyl) -3- (2-naphthyl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan was replaced by (E) - (3- (1- (3-chlorophenyl) -3- (5, 6,7, 8-tetrahydronaphthalen-2-yl) -1H-pyrazol-4-yl) acryloyl) -L-tryptophan, and the remaining desired starting materials, reagents and preparation method were the same as in comparative example 8 to give the product I-26-PROTAC.

I-26-PROTAC (comparative Compound 11)

¹ H NMR(600MHz，DMSO-d6)δ11.10(s，1H)，10.82-10.80(m，1H)，8.98(s，1H)，8.29(d，J＝8.2Hz，1H)，8.09(t，J＝5.7Hz，1H)，8.03(q，J＝3.3，2.2Hz，1H)，7.96(s，3H)，7.79(d，J＝7.3Hz，1H)，7.73(t，J＝8.8Hz，1H)，7.65-7.62(m，1H)，7.56-7.52(m，1H)，7.41(dd，J＝8.1，2.1Hz，1H)，7.33-7.30(m，2H)，7.27-7.22(m，1H)，7.19-7.14(m，1H)，7.10-6.99(m，2H)，6.97(t，J＝7.4Hz，1H)，6.58(t，J＝5.8Hz，1H)，6.53(d，J＝15.7Hz，1H)，5.17(dd，J＝12.9，5.4Hz，1H)，4.64(td，J＝8.5，5.1Hz，1H)，3.62-3.47(m，5H)，3.45-3.38(m，3H)，3.27-3.18(m，2H)，3.16-3.10(m，1H)，3.02-2.95(m，1H)，2.91-2.86(s，3H)，2.79-2.74(m，4H)，2.73-2.72(s，3H)，1.84-1.67(s，4H)；

Effect example 1 results of affinity test of partial Compounds with Bcl-xL, bcl-2 and Mcl-1

A BidBH3 polypeptide (amino acids 79-104: QEDIRNIARHLAQVGDSMDRSIPPG) with 26 amino acid residues was synthesized and labeled with 6-carboxyfluorescein succinimidyl ester (FAM) as fluorescent tag (FAM-Bid) at the N-terminus. Dissolving His-Bcl-xL protein or His-Bcl-2 protein or His-Mcl-1 protein and a small molecule compound to be detected in a phosphate buffer salt solution, incubating for 30 minutes at 37 ℃ in a dark place, then adding FAM-Bid polypeptide, mixing uniformly, and incubating for 20 minutes at 37 ℃ in a dark place. His-Bcl-XL protein, his-Bcl-2 protein, his-Mcl-1 protein and FAM-Bid polypeptide have final concentrations of 230nM,425nM,200nM and 10nM, respectively. The final concentrations of the compounds in the system were 1nM,10nM,100nM, 1. Mu.M, 10. Mu.M, 50. Mu.M and 100. Mu.M, respectively. 60 μl of each of the above reaction solutions was added to 384-well plates (three groups in parallel), and fluorescence polarization was immediately detected on an enzyme-labeled instrument. Fluorescence polarization (mP) was measured at 535nm emission wavelength resulting from 485nm wavelength excitation. Two control groups are simultaneously established, one control group is that the reaction system only contains Bcl-xL or Bcl-2 or Mcl-1 and FAM-Bid (equivalent to 0 percent inhibition rate), and the other control group is that the reaction system only contains FAM-Bid polypeptide. Protein inhibition was calculated from the control and the results of the measurement of the polarization values of the test compounds. Determination of IC by plotting the protein inhibition ratio against the log of the compound concentration ₅₀ Values. According to the formula ki= [ I]50/([L]50/Kd+[P]0/Kd+1) derivedCompetition inhibition constant Ki value for compound and protein. In the formula [ I ]]50 is the concentration of the compound at which the protein inhibition rate is 50% [ L ]]50 is the concentration of free FAM-Bid with 50% protein inhibition rate, kd is the dissociation constant of target protein and FAM-Bid polypeptide, [ P ]]0 is the free protein concentration at which the protein inhibition rate is 0%. The specific results are shown in Table 1.

TABLE 1 Bcl-X pair of compounds of the invention _L Competition inhibition constant K for Bcl-2 and Mcl-1 proteins _i

N.a. indicates inactivity.

The table shows that the phenylpyrazole compound provided by the invention can effectively and selectively inhibit the key protein MCL-1 in the apoptosis process at the molecular level.

Utility example 2 inhibition of proliferation activity of partial Compounds on several tumor cell lines

Human plasma cell leukemia cell H929 and human acute lymphoblastic leukemia cell RS4 to be detected; 11 with RPMI1640 medium containing 10% fetal bovine serum, human myelomonocytic leukemia cells MV-4-11 with IMDM medium containing 10% fetal bovine serum, human embryonic kidney 293T cells with DMEM medium containing 10% fetal bovine serum. Seeding cells in 96 well plates, the cell concentrations of H929 and MV-4-11 being 8000/100. Mu.L, RS4;11 cells at 16000/100. Mu.L and 293T cells at 10000/100. Mu.L; the zeroing group was supplemented with medium only. Compounds (five different concentrations 40. Mu.M, 20. Mu.M, 10. Mu.M, 5. Mu.M and 2.5. Mu.M, three groups in parallel at each concentration) were added separately to 96-well plates and then placed in CO at 37 ℃ ₂ Incubate in incubator for 48 hours. To each well, 10. Mu. LCCK-8 solution was added and incubated at 37℃for 2 hours, and the light absorption value of each well was measured at a wavelength of 450nm using a microplate reader. Calculate cell viability =(experimental group light absorption value-zeroing group light absorption value)/(control group light absorption value-zeroing group light absorption value). Determination of IC by plotting the log of the cell viability and the concentration of the compound ₅₀ Values. The specific results are shown in tables 2 and 3.

Table 2 Compounds of the invention are described in H929, MV-4-11, RS4; cytotoxicity test results on 11 and 293T cell lines

/: indicating no test data

The table shows that the phenylpyrazole compound provided by the invention has obvious killing effect and high selectivity on cancer cells, especially human plasma cell leukemia cell H929 and human myelomonocytic leukemia cell MV-4-11.

Utility example 3 Studies of the kinetics of Compound I-66 in living cells

MV-4-11 cells were seeded in 6-well cell culture plates with 1.2X106 cells per well, all incubated with 40. Mu.M I-66 treatment (5%DMSO). At the following time points (0, 1, 2, 3, 5, 7 and 8 h), one well of cells was collected and immediately centrifuged at 4℃and 200g for 5 minutes. The supernatant was analyzed by LC-MS/MS quantitative analysis as an extracellular component. The pelleted cells were lysed in RIPA lysis buffer (50mMTris,pH 7.4, 150mM NaCl,1%Triton X-100,0.1% SDS,1% sodium deoxycholate) for 30min. After cell lysis, the cells were centrifuged at 12000rpm for 20min, and the supernatant was collected as an intracellular component for LC-MS/MS analysis. The percentage of intracellular or extracellular compounds is calculated as follows:

The content of I-66 and its carboxylic acid form (I-67) in the intracellular and extracellular phases was accurately measured by incubating 40. Mu.M of I-66 with MV-4-11 cells at 0, 1, 2, 3, 5 and 8 hours, respectively. As shown in Table 4, I-66 and I-67 were not present in the cells at 0h and were fully exposed outside the cells. After 1h incubation of I-66 with cells, only I-67 was detected in the cells, accounting for 20% of the total weight. I-66 was not detected in the cell, confirming that I-66 was completely hydrolyzed to carboxylic acid I-67 after entering the cell to exert intracellular biological effects. As the treatment time of I-66 was prolonged, the weight of intracellular I-67 increased rapidly over time. The conversion of intracellular I-67 to extracellular I-67 was detected with apoptosis and death, resulting in a slow decrease in the ratio of I-67 in living cells.

TABLE 4 Table 4

The experiment shows that I-66 has better membrane permeability through dynamic monitoring of the amount of the compound in living cells, and is completely hydrolyzed into I-67 in the cells to exert biological functions.

Claims

1. A phenylpyrazole compound shown in a formula I, a stereoisomer or a pharmaceutically acceptable salt thereof,

wherein R is ^a Is R ¹ -(CH ₂ ) _m -；

M is 0, 1, 2, 3, 4, 5 or 6;

R ^c is H or C ₁ ～C ₄ An alkyl group;

R ^d is phenyl, 3-10 membered heteroaryl, substituted with one or more R ² Substituted phenyl or by one or more R ² Substituted 3-10 membered heteroaryl; the 3-10 membered heteroaryl and "substituted with one or more R ² In the 3-10 membered heteroaryl group in the substituted 3-10 membered heteroaryl group ", the number of heteroatoms is independently 1,2,3 or 4, and the heteroatoms are independently N, O and S; when a plurality of substituents are present, the substituents are the same or different;

said R is ² Independently is hydroxy or C ₁ ～C ₆ An alkyl group;

represents Z configuration, E configuration or a mixture thereof;

2. The phenylpyrazole compound according to claim 1 of formula I, its stereoisomers or pharmaceutically acceptable salts thereof, which fulfil one or more of the following conditions:

(1) Said R is ¹ To be covered by one or more R ^1-1 A substituted phenyl group;

(2) Said R is ^1-1 independently-NH ₂ 、-OH、C ₁ -C ₄ Alkoxy, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl;

Preferably, R ^1-1 independently-NH ₂ -OH or 3-8 membered cycloalkyl; alternatively, said R ^1-1 Independently C ₁ -C ₄ Alkoxy or 3-8 membered heterocycloalkyl;

(3) Said R is ^b1 、R ^b2 And R is ^b3 One of which is halogen or C ₁ ～C ₆ Alkyl, the rest is H; preferably, R ^b2 Is halogen or C ₁ ～C ₆ Alkyl, R ^b1 And R is ^b3 Is H; more preferably, R ^b2 Is halogen, R ^b1 And R is ^b3 Is H;

(4) Said R is ^d Is 3-10 membered heteroaryl, substituted with one or more R ² Substituted phenyl or by one or more R ² Substituted 3-10 membered heteroaryl; preferably, R ^d Is 3-10 membered heteroaryl;

(5) Said R is ² Is C ₁ ～C ₆ An alkyl group.

3. The phenylpyrazole compound according to claim 1 of formula I, its stereoisomers or pharmaceutically acceptable salts thereof, which fulfil one or more of the following conditions:

(1) M is 0, 1, 2, 3 or 4;

(2) Said R is ^1-1 The number of substitutions is 1, 2, 3, 4 or 5;

(3) Said R is ^1-1 Independently located ortho, meta or para to the "phenyl and pyrazole attachment site";

(4) When R is ^1-1 Is C ₁ -C ₄ Alkoxy, the C ₁ -C ₄ Alkoxy is methoxy, ethoxy, propoxy or butoxy;

(5) When R is ^1-1 In the case of 3-8 membered cycloalkyl, the 3-8 membered cycloalkyl is cyclopropyl, cyclopentyl, cyclohexyl or cycloheptyl;

(6) When R is ^1-1 In the case of 3-8 membered heterocycloalkyl, the 3-8 membered heterocycloalkyl is 5-6 membered heterocycloalkyl with 1-2 heteroatoms selected from N and/or O;

(7) When R is ^b1 、R ^b2 And R is ^b3 When independently halogen, the halogen is fluorine, chlorine, bromine or iodine;

(8) When R is ^b1 、R ^b2 And R is ^b3 Independently C ₁ ～C ₆ When alkyl, the C ₁ ～C ₆ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

(9) When R is ^c Is C ₁ ～C ₄ When alkyl, the C ₁ ～C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

(10) When R is ^d In the case of 3-10 membered heteroaryl, the 3-10 membered heteroaryl is a 5-10 membered heteroaryl with heteroatom selected from N and heteroatom number of 1-2;

(11) When R is ^d To be covered by one or more R ² When the 3-10 membered heteroaryl is substituted, the 3-10 membered heteroaryl is a 5-10 membered heteroaryl with heteroatom selected from N and heteroatom number of 1-2;

(12) Said R is ² The number of substitutions is 1, 2, 3, 4 or 5;

(13) When R is ² Is C ₁ ～C ₆ When alkyl, the C ₁ ～C ₆ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

4. The phenylpyrazole compound according to claim 1 of formula I, its stereoisomers or pharmaceutically acceptable salts thereof, which fulfil one or more of the following conditions:

(1) M is 0;

(2) Said R is ^1-1 The number of the substitutions is 1;

(3) Said R is ^1-1 Independently located para to the "phenyl and pyrazole attachment site";

(4) When R is ^1-1 Is C ₁ -C ₄ Alkoxy, the C ₁ -C ₄ Alkoxy is methoxy;

(5) When R is ^1-1 In the case of 3-8 membered cycloalkyl, the 3-8 membered cycloalkyl is cyclohexyl;

(6) When R is ^1-1 In the case of 3-8 membered heterocycloalkyl, the 3-8 membered heterocycloalkyl is morpholinyl;

(7) When R is ^b1 、R ^b2 And R is ^b3 When independently halogen, the halogen is chlorine;

(8) When R is ^d Is 3-10 membered heteroWhen aryl, the 3-10 membered heteroaryl is indolyl;

(9) When R is ^d To be covered by one or more R ² When substituted 3-10 membered heteroaryl, the 3-10 membered heteroaryl is indolyl;

(10) Said R is ² The number of substitutions was 1.

5. The phenylpyrazole compound according to claim 1, wherein the phenylpyrazole compound according to formula I, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof is according to scheme one or scheme two;

scheme one:

R ^a is R ¹ -(CH ₂ ) _m -；

M is 0;

said R is ¹ To be covered by one or more R ^1-1 A substituted phenyl group;

said R is ^1-1 independently-NH ₂ -OH or 3-8 membered cycloalkyl;

R ^c is H or C ₁ ～C ₄ An alkyl group;

R ^d is 3-10 membered heteroaryl or is substituted with one or more R ² Substituted 3-10 membered heteroaryl; the 3-10 membered heteroaryl and "substituted with one or more R ² In the 3-10 membered heteroaryl group in the substituted 3-10 membered heteroaryl group ", the number of hetero atoms is independently 1 or 2, and the hetero atoms are N;

the R is ² Is C ₁ ～C ₆ An alkyl group;

scheme II:

R ^a is R ¹ -(CH ₂ ) _m -；

M is 0;

said R is ¹ To be covered by one or more R ^1-1 A substituted phenyl group;

said R is ^1-1 Independently C ₁ -C ₄ Alkoxy or 3-8 membered heterocycloalkyl; in the 3-8 membered heterocycloalkylThe number of heteroatoms of (a) is independently 1,2,3 or 4, and the heteroatoms are independently N, O and S;

R ^c is H or C ₁ ～C ₄ An alkyl group;

the R is ² Is C ₁ ～C ₆ An alkyl group.

6. The phenylpyrazole compound according to claim 1 of formula I, its stereoisomers or pharmaceutically acceptable salts thereof, which fulfil one or more of the following conditions:

(1) The R is ^a Is that

(2) The R is ^d Is that

(3)Is->

7. The phenylpyrazole compound according to claim 1, which is represented by the formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the phenylpyrazole compound is any one of the following:

8. A compound of formula II, a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

wherein,,

x is

L is

The end a is connected with Y, and the end b is connected with X;

n1 and n2 are independently 0, 1,2,3, 4 or 5;

L ¹ and L ² Independently is-O- -S-or-NH-;

y is

R ^b1 、R ^b2 、R ^b3 、R ^c And R is ^d Is as defined in any one of claims 1 to 7;

L ³ is-R ³ -(CH ₂ ) _m -；-(CH ₂ ) _m -terminal to pyrazole;

m is 0, 1,2,3, 4, 5 or 6;

said R is ³ Is phenyl or is substituted by one or more R ^3-1 A substituted phenyl group; when a plurality of substituents are present, the substituents are the same or different;

l is connected to L ³ Ortho, meta or para to the pyrazole attachment site;

represents Z configuration, E configuration or a mixture thereof;

9. A compound of formula II, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as claimed in claim 8 which satisfies one or more of the following conditions:

(1) The L is

(2) The L is ¹ is-O-;

(3) The L is connected to L ³ Para to the pyrazole attachment site;

(4) Said R is ³ Is phenyl;

(5) The R is ^3-1 independently-NH ₂ 、-OH、C ₁ -C ₄ Alkoxy, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl; the number of heteroatoms in the 3-8 membered heterocycloalkyl is 1,2,3 or 4, and the heteroatoms are N, O and S independently; preferably, said R ^3-1 independently-NH ₂ -OH or 3-8 membered cycloalkyl.

10. A compound of formula II, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as claimed in claim 8 which satisfies one or more of the following conditions:

(1) N1 is 0, 1,2 or 3;

(2) Said m is 0, 1,2,3 or 4; preferably, m is 0;

(3) Said R is ^3-1 The number of substitutions is 1,2,3, 4 or 5;

(4) When R is ^3-1 Is C ₁ -C ₄ Alkoxy, the C ₁ -C ₄ Alkoxy is methoxy, ethoxy, propoxy or butoxy, preferably methoxy;

(5) When R is ^3-1 In the case of a 3-8 membered cycloalkyl group, the 3-8 membered cycloalkyl group is cyclopropyl, cyclopentyl, cyclohexyl or cycloheptyl, preferably cyclohexyl;

(6) When R is ^3-1 In the case of 3-8 membered heterocycloalkyl, the 3-8 membered heterocycloalkyl is a 5-6 membered heterocycloalkyl having 1 to 2 heteroatoms selected from N and/or O, for example morpholinyl.

11. A compound of formula II, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as claimed in claim 8 which satisfies one or more of the following conditions:

(1) The X is

(2) The L is

(3) Y is

12. The compound of formula II, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to claim 8, which is a compound of formula II-a as follows

Wherein R is ^c 、R ^b1 、R ^b2 、R ^b3 And R is ^d Is as defined in any one of claims 8 to 11;

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

13. A compound of formula II, a stereoisomer thereof or a pharmaceutically acceptable salt thereof according to any one of claims 8 to 11, which is any one of the following:

14. a process for the preparation of a compound of formula I according to any one of claims 1 to 7, comprising process 1 or process 2;

wherein R is ^a 、R ^b1 、R ^b2 、R ^b3 And R is ^d Is as defined in any one of claims 1 to 7;

method 2, comprising the steps of: in organic solvent, "R" is used ^c Is C ₁ ～C ₄ The compound I of the alkyl group "is subjected to hydrolysis reaction as shown below to give" R " ^c The compound I is H';

the reaction conditions of the method 1 and the method 2 preferably satisfy one or more of the following conditions:

(1) In the method 1, the organic solvent is an amide solvent, such as N, N-dimethylformamide;

(2) In the method 1, the catalyst is one or more of N, N ' -diisopropylethylamine, 2- (7-benzotriazol-oxide) -N, N, N ', N ' -tetramethylurea hexafluorophosphate and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride;

(3) In the method 1, the mol ratio of the compound III to the compound IV is 1:1-1:2;

(4) In the method 2, the organic solvent is an ether solvent, such as tetrahydrofuran;

(5) In process 2, the hydrolysis is carried out in the presence of a base, such as an alkali metal hydroxide, e.g., naOH.

15. The method of preparing a compound of formula I according to claim 14, further comprising the steps of:

wherein R is ^a 、R ^b1 、R ^b2 And R is ^b3 The method of claim 14; r is R ^e Is C ₁ ～C ₄ An alkyl group;

step one: in an organic solvent, carrying out condensation reaction on a compound VIII and a compound IX in the presence of a catalyst to obtain the compound VII;

step two: in an organic solvent, in the presence of a catalyst, carrying out a Wilsmeier-Hakk formylation reaction shown by a compound VII and disubstituted formamide to obtain the compound V;

step three: in an organic solvent, in the presence of alkali, carrying out an addition elimination reaction shown by a compound IV and a compound V to obtain the compound III;

preferably, in step one it fulfils one or more of the following conditions:

(1) The organic solvent is an alcohol solvent, such as ethanol;

(2) The catalyst is acetic acid;

preferably, in step two it fulfils one or more of the following conditions:

(1) The organic solvent is formamide solvent, such as N, N-dimethylformamide;

(2) The disubstituted formamide is N, N-dimethylformamide;

(3) The catalyst is POCl ₃ ；

(4) The molar ratio of said compound VII to said catalyst is from 1:2 to 1:5, for example 1:4;

preferably, in step three it fulfils one or more of the following conditions:

(1) The R is ^e Is ethyl;

(2) The organic solvent is an ether solvent, such as tetrahydrofuran;

(3) The molar ratio of the compound V to the compound VI is from 1:2 to 1:3, for example 1:2.2.

16. A process for the preparation of a compound of formula II according to any one of claims 12 to 13, comprising the steps of: in a solvent, under the action of a catalyst, reacting the compound II-b with the compound II-c to obtain a compound shown as a formula II-a;

wherein R is ^b1 、R ^b2 、R ^b3 、R ^c And R is ^d Is as defined in any one of claims 8 to 13;

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

the conditions of the reaction preferably satisfy one or more of the following conditions:

(1) The solvent is formamide solvent, such as N, N-dimethylformamide;

(2) The catalyst is N, N' -diisopropylethylamine;

(3) The molar ratio of the compound II-b to the compound II-c is 1:11-1:1.5.

17. The method of preparing a compound of formula II according to claim 16, further comprising the steps of:

wherein R is ^b1 、R ^b2 、R ^b3 、R ^c And R is ^d Is defined as set forth in claim 16;

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

step one: in a solvent, reacting the compound II-e with the compound II-f in the presence of alkali and a condensation reagent to obtain a compound shown as a formula II-d;

Step two: removing protecting groups from the compound II-d in a solvent to obtain a compound shown in a formula II-b;

preferably, in step one it fulfils one or more of the following conditions:

(1) The solvent is formamide solvent, such as N, N-dimethylformamide;

(2) The catalyst is N, N ' -diisopropylethylamine and 2- (7-benzotriazol-N, N, N ', N ' -tetramethylurea hexafluorophosphate;

(3) The molar ratio of the compound of formula II-e to the compound II-f is 1:1 to 1:1.5, preferably 1:1.1 to 1:1.2;

preferably, in step two it fulfils one or more of the following conditions:

(1) The solvent is halogenated hydrocarbon solvent such as dichloromethane;

(2) The deprotecting reagent is an acid reagent such as trifluoroacetic acid.

18. A pharmaceutical composition comprising substance a, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutical adjuvant; substance a is a compound of formula I as defined in any one of claims 1 to 7 or a compound of formula II as defined in any one of claims 8 to 13.

19. Use of substance a, a stereoisomer thereof or a pharmaceutically acceptable salt thereof for the preparation of a BCL-2 anti-apoptotic protein inhibitor or for the preparation of a medicament for the treatment and/or prophylaxis of a disease associated with BCL-2 anti-apoptotic proteins;

Substance a is a compound of formula I as defined in any one of claims 1 to 7 or a compound of formula II as defined in any one of claims 8 to 13;

the "BCL-2 anti-apoptotic protein" may be BCL-X _L One or more of BCL-2 and MCL-1 proteins, preferably MCL-1;

the "disease associated with BCL-2 anti-apoptotic proteins" may be cancer; the cancer is preferably leukemia; such as human acute lymphoblastic leukemia or human plasma cell leukemia.

20. The use of substance B, a stereoisomer thereof or a pharmaceutically acceptable salt thereof in the preparation of a protease degradation agent;

substance B is a compound of formula II as defined in any one of claims 8 to 13.