WO2022143576A1

WO2022143576A1 - Pyrazoloquinazoline compound, and preparation method therefor and use thereof

Info

Publication number: WO2022143576A1
Application number: PCT/CN2021/141831
Authority: WO
Inventors: 魏钊; 王文义; 崔洪; 李元念; 周红; 石峰
Original assignee: 恒元生物医药科技(苏州)有限公司
Priority date: 2020-12-31
Filing date: 2021-12-28
Publication date: 2022-07-07
Also published as: CN116528871A; TW202241413A

Abstract

The present invention relates to a pyrazoloquinazoline compound, and a preparation method therefor and the use thereof. The present invention particularly relates to a pyrazoloquinazoline compound as shown in formula I, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of the pharmaceutically acceptable salt thereof. The compound has an obvious inhibitory effect on PLK1.

Description

A kind of pyrazoloquinazoline compound, its preparation method and application

technical field

The invention relates to a pyrazoloquinazoline compound, a preparation method and application thereof.

Background technique

PLKs (Polo-like kinases) are serine/threonine kinases. So far, five subtypes have been found in their family, namely PLK1-5. PLKs are mainly expressed in dividing cells and play a crucial role in cell cycle control and mitosis. In the PLK family, PLK1 is the most studied and the most detailed, and its biological activity is also the most clearly described. PLK1 is widely recognized by the scientific community as a key kinase that directs mitotic entry, centrosome maturation and separation, bipolar spindle formation, metaphase to anaphase transition, and initiation of cytoplasmic division. Studies have found that PLK1 is not only highly expressed in various tumor tissues, but also activated during DNA damage and replication stress, which is also consistent with the abnormally active tumor cell proliferation. Knockout of the PLK1 gene by chemical small molecule inhibitors or biological means can find that a large number of tumor cells stay in the G2/M phase, and further lead to tumor cell apoptosis. Therefore, PLK1 is considered to be a promising anti-tumor target, especially its high expression in various tumor tissues makes it possible to have broad-spectrum anti-tumor potential. In addition, the expression level of PLK1 in normal cells is low, so the development of PLK1 inhibitors as antitumor drugs may have a good safety window.

Compared with the high expression of PLK1 in tumor cells, PLK2 and PLK3 are more expressed in post-mitotic cells such as nerve cells, so the development of PLK1-specific inhibitors may avoid the toxic and side effects caused by simultaneous inhibition of PLK2/3. The PLK1 inhibitor Volasertib (BI-6727) developed by BI (Boehringer Ingelheim) has serious toxic and side effects in the clinic due to its lack of selectivity for PLK2/3, which limits its efficacy, thus leading to its phase III clinical failure. As a PLK1-specific inhibitor, Onvansertib, a small molecule inhibitor developed by Cardiff Oncology, has shown good antitumor activity and controllable side effects in multiple proof-of-concept Phase II clinical trials. However, Onvansertib still has much room for improvement, such as poor permeation properties resulting in lower bioavailability. Moreover, due to its long half-life (t _1/2 is greater than 24 hours), it can only be administered intermittently during the treatment cycle, which further limits its therapeutic effect in addition to obvious toxic and side effects. .

SUMMARY OF THE INVENTION

The technical problem to be solved by the present invention is the defects of single PLK1 inhibitor structure and poor inhibitory effect in the prior art, and provides a pyrazoloquinazoline compound, its preparation method and application, the compound inhibits PLK1 The effect is obvious.

The present invention solves the above-mentioned technical problems through the following technical solutions.

The present invention provides a pyrazoloquinazoline compound as shown in formula I, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof;

Wherein, R ¹ is C ₁ -C ₄ alkoxy, C ₁ -C ₄ alkoxy substituted by one or more R ^1-1 or 3-6 membered cycloalkyloxy; R ^1-1 is independently It is halogen or 3-6 membered cycloalkane;

R ² is H, halogen or C ₁ -C ₄ alkyl;

R ³ is a 5- to 10-membered heterocycloalkyl group or a 5- to 10-membered heterocycloalkyl group substituted by one or more R ^3-1 ; in the 5- to 10-membered heterocycloalkyl group, the heteroatom is selected from N One or more of , O and S, the number of heteroatoms is 1-2; R ^3-1 is independently C ₁ -C ₄ alkyl;

R ⁴ is cyano, 3-6 membered cycloalkane, 3-6 membered cycloalkane substituted by one or more R ^4-1 , 3-6 membered heterocycloalkane or substituted by one or more R ^4-2 3-6-membered heterocycloalkane; C ₁ -C ₄ alkyl substituted by one or more R ^4-3 ; in the 3-6 membered heterocycloalkyl, the heteroatom is selected from N, O and S One or more of , the number of heteroatoms is 1-2; R ^4-1 is independently halogen, hydroxyl or C ₁ -C ₄ alkyl; R ^4-2 is independently halogen or C ₁ -C ₄ alkyl; R ^4-3 is independently halogen, cyano or hydroxyl;

R ⁵ is H, hydroxy, C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy.

In a certain scheme, in the pyrazoloquinazoline compound, its pharmaceutically acceptable salt, its solvate or its pharmaceutically acceptable salt solvate, certain groups is defined as follows, and the remaining groups are defined as described in any one of the schemes above (hereinafter referred to as "in a scheme"):

In one embodiment, R ^1-1 is independently F, Cl, Br or I, eg, F.

In one embodiment, R ^1-1 is independently a 3- to 6-membered cycloalkane, such as cyclopropane, cyclobutane, or cyclopentane.

In a certain scheme, when R ¹ is C ₁ -C ₄ alkoxy, the C ₁ -C ₄ alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, For example methoxy.

In a certain scheme, when R ¹ is C ₁ -C ₄ alkoxy substituted by one or more R ^1-1 , the C ₁ -C ₄ alkoxy may be methoxy, ethoxy , isopropoxy or tert-butoxy, for example methoxy.

In a certain scheme, when R ¹ is 3-6 membered cycloalkyloxy, the 3-6 membered cycloalkoxy is cyclopropyloxy, cyclobutyloxy or cyclopentyloxy , such as cyclopropyloxy or cyclobutyloxy.

In one scheme, R ¹ can be

E.g

^In one embodiment, when R2 is halogen, the halogen can be F, Cl, Br or I.

In a certain scheme, when R ² is a C ₁ -C ₄ alkyl group, the C ₁ -C ₄ alkyl group can be methyl, ethyl, n-propyl, isopropyl, n-butyl, isopropyl Butyl or tert-butyl, eg methyl.

In a certain scheme, when R ^3-1 is a C ₁ -C ₄ alkyl group, the C ₁ -C ₄ alkyl group can be methyl, ethyl, n-propyl, isopropyl, n-butyl , isobutyl or tert-butyl such as methyl.

In a certain scheme, when R ³ is a 5-10-membered heterocycloalkyl substituted by one or more R ^3-1 , the 5-10-membered heterocycloalkyl can be a 6-9 membered heterocycle Alkyl, such as 6- to 9-membered heterocycloalkyl containing two N, and piperazinyl, hexahydropyridazinyl, hexahydropyrimidinyl, 3,8-diazabicyclo[3.2.1]octane base, octahydropyrrole[1,2-a]pyrazinyl, 2,6-diazaspiro[3.4]octyl, 3,6-diazabicyclo[3.2.0]heptyl, 1, 6-diazaspiro[3.4]octyl or octahydropyrrole[3,4-c]pyrrolyl.

In a certain scheme, R ³ is preferably

(b is with

junction site).

In a certain scheme, when R ³ can be

In a certain scheme, R ³ can be

In a certain scheme, R ⁴ is cyano, 3-6 membered cycloalkane substituted with one or more R ^4-1 , 3-6 membered heterocycloalkane, or 3-membered cycloalkane substituted with one or more R ^4-2 ~6-membered heterocycloalkane.

In one embodiment, R ^4-1 is halo or hydroxy.

In one embodiment, R ^4-2 is C ₁ -C ₄ alkyl.

^In one embodiment, R5 is H.

In one embodiment, when ^R4-1 is halogen, the halogen is F, Cl, Br or I, eg, F.

In a certain scheme, when R ^4-1 is a C ₁ -C ₄ alkyl group, the C ₁ -C ₄ alkyl group can be methyl, ethyl, n-propyl, isopropyl, n-butyl , isobutyl or tert-butyl such as methyl or ethyl.

In a certain scheme, when R ^4-2 is a C ₁ -C ₄ alkyl group, the C ₁ -C ₄ alkyl group can be methyl, ethyl, n-propyl, isopropyl, n-butyl , isobutyl or tert-butyl such as methyl or ethyl.

In one embodiment, when ^R4-2 is halogen, the halogen is F, Cl, Br or I, eg, F.

In a certain scheme, when R ⁴ is a 3-6 membered cycloalkane, the 3-6 membered cycloalkane can be cyclopropane, cyclobutane or cyclopentane, such as cyclopropane or cyclobutane.

In a certain scheme, when R ⁴ is a 3-6 membered cycloalkane substituted by one or more R ^4-1 , the 3-6 membered cycloalkane can be cyclopropane, cyclobutane or cyclopentane , such as cyclopropane or cyclobutane.

In a certain scheme, when R ⁴ is a 3-6 membered cycloalkane substituted by one or more R ^4-1 , the 3-6 membered cycloalkane substituted by one R ^4-1 may be

In a certain scheme, when R ⁴ is a 3-6 membered heterocycloalkane, the 3-6 membered heterocycloalkane may be

In a certain scheme, when R ⁴ is a 3-6 membered heterocycloalkane substituted with one R ^4-2 , the 3-6 membered heterocycloalkane can be a 4-membered heterocycloalkane containing one N or O or a 5-membered heterocycloalkane containing one N or O.

In a certain scheme, when R ⁴ is a 3-6 membered heterocycloalkane substituted by one or more R ^4-2 , the 3-6 membered heterocycle substituted by one or more R ^4-2 Alkanes are

In a certain scheme, when R ⁴ is C ₁ -C ₄ alkyl substituted by one or more R ^4-3 , said C ₁ -C ₄ alkane substituted by one or more R ^4-3 base for

In a certain scheme, R ⁴ is cyano, 3-6 membered cycloalkane substituted with one or more R ^4-1 , 3-6 membered heterocycloalkane, or 3-membered cycloalkane substituted with one or more R ^4-2 ~6-membered heterocycloalkane, or C ₁ -C ⁴ alkyl substituted with one or more R ^4-3 _.

In one embodiment, R ⁴ is a 3-6 membered cycloalkane substituted with one or more R ^4-1 , wherein R ^4-1 is halo or hydroxy.

In one embodiment, R ⁴ is a 3-6 membered heterocycloalkane substituted with one or more R ^4-2 , wherein R ^4-2 is a C ₁ -C ⁴ alkyl group _.

In one embodiment, R ⁴ is C ₁ -C ₄ alkyl substituted with one or more R ^4-3 , wherein R ^4-3 is hydroxy.

^In a certain scheme, R is selected from the group consisting of cyano,

^In one embodiment, R5 is H or hydroxy.

In a certain scheme, R ⁵ is a C ₁ -C ₄ alkyl group, and the C ₁ -C ₄ alkyl group can be methyl, ethyl, isopropyl or tert-butyl, such as methyl or ethyl.

In a certain scheme, R ⁵ is C ₁ -C ₄ alkoxy, and the C ₁ -C ₄ alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, such as methyl Oxygen

The present invention also provides a pyrazoloquinazoline compound as shown in formula Ia, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof:

In a certain aspect, the present invention provides a pyrazoloquinazoline compound, a pharmaceutically acceptable salt thereof, a solvate or a solvate of a pharmaceutically acceptable salt thereof as shown in any one of the following:

The present invention also provides a pyrazoloquinazoline compound represented by formula II, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof:

wherein, R ¹ is C ₁ -C ₄ alkoxy substituted by one or more R ^1-1 ; R ^1-1 is independently halogen;

R ² is H, halogen or C ₁ -C ₄ alkyl;

R ^3' is a 7-9 membered heterocycloalkyl or a 7-9 membered heterocycloalkyl substituted by one R ^3-1 ; the 7-9 membered heterocycloalkyl is a 7-9 membered heterospirocycloalkane or 7- to 9-membered hetero-bridged cycloalkyl; the 7- to 9-membered heterocycloalkyl substituted by one R ^3-1 is a 7- to 9-membered heterospirocycloalkyl substituted by one R ^3-1 or A 7- to 9-membered hetero-bridged cycloalkyl substituted by one R ^3-1 ; the heteroatom is selected from one or more of N, O and S, and the number of heteroatoms is 1-2; R ^3-1 is C ₁ -C ₄ alkyl;

R ^4' is independently C ₁ -C ₄ alkyl substituted with one R ^4-3 ; R ^4-3 is hydroxy or halogen;

R ⁵ is H, hydroxy C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy.

In one embodiment, R ^1-1 is F, Cl, Br or I, eg F.

In a certain scheme, when R ¹ is C ₁ -C ₄ alkoxy substituted by one or more R ^1-1 , the

In one embodiment, ^R3-1 can be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, eg methyl.

In a certain scheme, when R ^3' is a 7- to 9-membered heterospirocycloalkyl group, the 7- to 9-membered heterospirocycloalkyl group can be a 7- to 9-membered heterospirocycloalkane containing two nitrogen atoms base, e.g.

In a certain scheme, when R ^3' is a 7-9 membered heterospirocycloalkyl substituted by one R ^3-1 , the 7-9 membered heterospirocyclic substituted or unsubstituted by one R ^3-1 The alkyl group may be a 7- to 9-membered heterospirocycloalkyl group substituted with one R ^3-1 containing two nitrogen atoms, such as

In a certain scheme, when R ^3' is a 7- to 9-membered hetero-bridged cycloalkyl group, the 7- to 9-membered hetero-bridged cycloalkyl group can be a 7- to 9-membered hetero-bridged cycloalkyl group containing two Ns ,E.g

In a certain embodiment, when R ^3' is a 7-9 membered heterobridged cycloalkyl substituted by one R ^3-1 , the 7-9 membered heterobridged cycloalkyl substituted by one R ^3-1 can be is a 7- to 9-membered heterobridged cycloalkyl substituted with one R ^3-1 containing two nitrogens, such as

In one embodiment, when ^R4-3 is halogen, the halogen is F, Cl, Br or I.

In a certain scheme, when R ^4' is independently C ₁ -C ₄ alkyl substituted with one R ^4-3 , the C ₁ -C ₄ alkyl may be methyl, ethyl, n-propyl , isopropyl, n-butyl, isobutyl or tert-butyl, eg methyl.

^In one aspect, R4 can be _-CH2OH .

In a certain scheme, when R ⁵ is a C ₁ -C ₄ alkyl group, the C ₁ -C ₄ alkyl group can be methyl, ethyl, isopropyl or tert-butyl, such as methyl or ethyl base.

In a certain scheme, when R ⁵ is C ₁ -C ₄ alkoxy, the C ₁ -C ₄ alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, For example methoxy.

In a certain scheme, the pyrazoloquinazoline compound represented by formula II, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof may be as follows Either shown:

The present invention also provides a pyrazoloquinazoline compound represented by formula III, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof:

Wherein, R ¹ is C ₁ -C ₄ alkoxy or C ₁ -C ₄ alkoxy substituted by one or more R ^1-1 ; R ^1-1 is independently halogen or 3-6 membered cycloalkane;

R ² is H, halogen or C ₁ -C ₄ alkyl;

R ⁵ is H, hydroxy, C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy.

In one embodiment, R ¹ is a C ₁ -C ₄ alkoxy group, such as methoxy, ethoxy, isopropoxy or tert-butoxy.

In a certain scheme, R ¹ is C ₁ -C ₄ alkoxy substituted by one or more R ^1-1 , wherein the C ₁ -C ₄ alkoxy may be methoxy, ethoxy, Isopropoxy or tert-butoxy, eg methoxy.

In one embodiment, R ¹ is C ₁ -C ₄ alkoxy substituted with one or more R ^1-1 , wherein R ^1-1 is independently F, Cl, Br, or I, eg, F.

In a certain scheme, R ¹ can be selected from

^In one embodiment, R2 is H.

^In a certain embodiment, R2 is halogen which is F, Cl, Br or I.

In one embodiment, R ² is a C ₁ -C ₄ alkyl group such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

In a certain scheme, R ³ is a 5- to 10-membered heterocycloalkyl, such as a 6- to 9-membered heterocycloalkyl, such as a 6- to 9-membered heterocycloalkyl containing two Ns, and another example is piperazinyl, six-membered heterocycloalkyl Hydropyridazinyl, hexahydropyrimidinyl, 3,8-diazabicyclo[3.2.1]octyl, octahydropyrrole[1,2-a]pyrazinyl, 2,6-diazaspiro[ 3.4]Octyl, 3,6-diazabicyclo[3.2.0]heptyl, 1,6-diazaspiro[3.4]octyl or octahydropyrrole[3,4-c]pyrrolyl .

In one embodiment, R ³ is a 5-10 membered heterocycloalkyl substituted with one or more R ^3-1 , wherein the 5-10 membered heterocycloalkyl is, for example, a 6-9 membered heterocycloalkyl , such as a 6- to 9-membered heterocycloalkyl group containing two Ns, and another example, piperazinyl, hexahydropyridazinyl, hexahydropyrimidinyl, 3,8-diazabicyclo[3.2.1]octyl, Octahydropyrrole[1,2-a]pyrazinyl, 2,6-diazaspiro[3.4]octyl, 3,6-diazabicyclo[3.2.0]heptyl, 1,6- Diazaspiro[3.4]octyl or octahydropyrrole[3,4-c]pyrrolyl, wherein R ^3-1 is C ₁ -C ₄ alkyl, such as methyl, ethyl, n-propyl, isopropyl propyl, n-butyl, isobutyl or tert-butyl.

In one scheme, R ³ is

In one scheme, R ³ is

In one scheme, R ³ is

^In one embodiment, R4 is cyano.

In one embodiment, R4 is a 3-6 ^membered cycloalkane, such as cyclopropane, cyclobutane or cyclopentane.

In a certain scheme, R ⁴ is a 3-6 membered cycloalkane substituted by one or more R ^4-1 , wherein the 3-6 membered cycloalkane is, for example, cyclopropane, cyclobutane or cyclopentane, R ^4-1 is halogen such as F, Cl, Br or I.

In a certain scheme, R ⁴ is a 3-6 membered cycloalkane substituted by one or more R ^4-1 , wherein the 3-6 membered cycloalkane is, for example, cyclopropane, cyclobutane or cyclopentane, R ^4-1 is a C ₁ -C ₄ alkyl group, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

In one scheme, R ⁴ is

In one embodiment, R ⁴ is a 3-6 membered heterocycloalkane, such as a 4-membered heterocycloalkane containing one N or O or a 5-membered heterocycloalkane containing one N or O.

In a certain scheme, R ⁴ is a 3-6 membered heterocycloalkane substituted with one or more R ^4-2 , such as a 4-membered heterocycloalkane containing one N or O or a 5-membered heterocycloalkane containing one N or O Cycloalkanes wherein R ^4-2 is halogen such as F, Cl, Br or I.

In one scheme, R ⁴ is

In a certain scheme, R ⁴ is a 3-6 membered heterocycloalkane substituted with one or more R ^4-2 , such as a 4-membered heterocycloalkane containing one N or O or a 5-membered heterocycloalkane containing one N or O Cycloalkane, wherein R ^4-2 is a C ₁ -C ₄ alkyl group such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

In one embodiment, R ⁴ is C ₁ -C ₄ alkyl substituted with one or more R ^4-3 , wherein the C ₁ -C ₄ alkyl is methyl, ethyl, isopropyl or tertiary Butyl, R ^4-3 is halogen, such as F, Cl, Br or I.

In one embodiment, R ⁴ is C ₁ -C ₄ alkyl substituted with one or more R ^4-3 , wherein the C ₁ -C ₄ alkyl is methyl, ethyl, isopropyl or tertiary Butyl, R ^4-3 is cyano.

In one embodiment, R ⁴ is C ₁ -C ₄ alkyl substituted with one or more R ^4-3 , wherein the C ₁ -C ₄ alkyl is methyl, ethyl, isopropyl or tertiary Butyl, R ^4-3 is hydroxy.

In one scheme, R ⁴ is

^In one embodiment, R5 is H or hydroxy.

In a certain embodiment, R5 is _C1 ^- _C4 alkyl, such as methyl, ethyl, isopropyl or tert-butyl.

In one embodiment, R ⁵ is C ₁ -C ₄ alkoxy, such as methoxy, ethoxy, isopropoxy or tert-butoxy.

In a certain scheme, the described pyrazoloquinazoline compounds as shown in formula III are:

The present invention also provides a preparation method of a pyrazoloquinazoline compound shown in formula I, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof, The method comprises the following steps: in an organic solvent, in an alkali reagent, under the action of a palladium catalyst, compound 1g, a ligand reagent and R ³ -H are subjected to a coupling reaction to obtain compound I;

Wherein, R ¹ , R ² , R ³ , R ⁴ or R ⁵ are as defined above; X is halogen; L is hydrochloride.

In the coupling reaction, the palladium catalyst may be tridibenzylideneacetone dipalladium.

In the coupling reaction, the organic solvent can be a conventional solvent for this type of reaction in the art, such as tetradioxane.

In the coupling reaction, the base reagent may be cesium carbonate.

In the coupling reaction, the ligand reagent may be 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene.

The present invention also provides a pyrazoloquinazoline compound represented by formula II, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof, comprising the following In the above steps, in an organic solvent, under the action of a palladium catalyst, compound 1g, a ligand reagent and R ³ -L are subjected to a coupling reaction to obtain compound II;

Wherein, R ¹ , R ² , R ^3' , R ^4' or R ⁵ are as defined above; X is halogen; L is hydrochloride.

In the coupling reaction, the base can be cesium carbonate.

The present invention provides the above-mentioned pyrazoloquinazoline compound represented by formula I, its pharmaceutically acceptable salt, its solvate or its solvate of pharmaceutically acceptable salt in Application in the preparation of PLK1 inhibitor; the inhibitor is preferably an inhibitor used in vitro.

The present invention also provides a pharmaceutical composition, the pharmaceutical composition further comprises a pyrazoloquinazoline compound shown in formula I, a pharmaceutically acceptable salt thereof, a solvate thereof or a pharmaceutically acceptable compound thereof Solvates of acceptable salts and pharmaceutical excipients.

The present invention also provides a pyrazoloquinazoline compound represented by formula I, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof as described above in application in the preparation of medicines;

The medicament is a medicament for treating at least one of the following diseases: breast cancer, prostate cancer, lung cancer, colorectal cancer, liver cancer, pancreatic cancer, gastric cancer, esophageal cancer, melanoma, multiple myeloma, leukemia and lymphoma.

The present invention provides a pyrazoloquinazoline compound as shown in formula II as described above, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof in the preparation of Use in PLK1 inhibitor; the inhibitor is preferably an inhibitor used in vitro.

The present invention also provides a pharmaceutical composition comprising a pyrazoloquinazoline compound as shown in formula I, a pharmaceutically acceptable salt thereof, a solvate thereof or a pharmaceutically acceptable compound thereof Solvates and pharmaceutical excipients of the accepted salts.

The present invention also provides the above-mentioned pyrazoloquinazoline compound represented by formula II, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof application in the preparation of medicines;

The medicine is a medicine for treating at least one of the following diseases: breast cancer, prostate cancer, lung cancer, colorectal cancer, liver cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, melanoma, osteosarcoma, multiple myeloma , leukemia and lymphoma.

The present invention also provides a pharmaceutical composition comprising a compound of the present invention, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof, and one or more chemotherapeutics agent.

In one embodiment, the one or more chemotherapeutic agents are concurrently, separately or consecutively with a compound of the invention, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof use.

Terminology Explanation

In various parts of this specification, the substituents of the compounds disclosed in the present invention are disclosed in terms of group type or scope. Specifically, the present invention includes each and every independent subcombination of each member of these group species and ranges. For example, the term " _C1 - _C4 alkyl" specifically refers to the independently disclosed methyl, ethyl, C3 alkyl (ie _propyl , including n-propyl and isopropyl), _C4 alkyl (ie butyl) , including n-butyl, isobutyl, sec-butyl and tert-butyl).

When a listed substituent does not indicate through which atom it is attached to a compound included in the general formula but not specifically mentioned, such substituent may be bonded through any of its atoms. Combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

When a group is enumerated without expressly indicating that it has a substituent, such group is only meant to be unsubstituted. For example, when there is no definition of "substituted or unsubstituted" before "C ₁ -C ₄ alkyl", it refers only to "C ₁ -C ₄ alkyl" itself or "unsubstituted C ₁ -C ₄ alkyl".

The term "C _i -C _j " denotes a range of carbon numbers, where i and j are integers and j is greater than i, and the range of carbon numbers includes the endpoints (ie, i and j) and every integer point between the endpoints. For example, _C1 - _C6 represents the range of 1 to 6 carbon atoms, including 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms and 6 carbon atoms. In some embodiments, the term "C ₁ -C ₁₂ " means 1 to 12, especially 1 to 10, especially 1 to 8, especially 1 to 6, especially 1 to 5, especially 1 to 4, especially 1 to 3, or specifically 1 to 2 carbon atoms.

The term "alkyl" refers to a saturated straight or branched chain hydrocarbon group. The term "Ci- _Cj alkyl" refers to an alkyl group having _i to j carbon atoms. In some embodiments, the alkyl group contains 1 to 12 carbon atoms. In some embodiments, the alkyl group contains 1 to 11 carbon atoms, 1 to 10 carbon atoms, 1 to 9 carbon atoms, 1 to 8 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms , 1 to 5 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, 1-propyl (n-propyl), 2-propyl (isopropyl), 1-butyl (n-butyl), 2-methyl-1 -propyl (isobutyl), 2-butyl (neobutyl), 2-methyl-2-propyl (tert-butyl), 1-pentyl (n-pentyl), 2-pentyl, 3 -pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2- Hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl -3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, 1-heptyl, 1-octyl, etc.

The term "alkoxy" refers to an alkyl group, as defined above, attached to the parent molecule through an oxygen atom. The term "Ci-Cjalkoxy" means that the alkyl portion of the alkoxy group has _i to _j carbon atoms. In some embodiments, the alkoxy group contains 1 to 12 carbon atoms. In some embodiments, the alkoxy group contains 1 to 11 carbon atoms. In some embodiments, the alkoxy group contains 1 to 10 carbon atoms, 1 to 9 carbon atoms, 1 to 8 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms.

The term "cycloalkane" or "cycloalkyl" refers to a saturated monocyclic cyclic group consisting of only carbon atoms having the specified number of carbon atoms (eg, C3 _- _C6 ). Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

The term "heterocycloalkyl" refers to a specified number of ring atoms (eg, 5-10 members), a specified number of heteroatoms (eg, 1, 2, or 3), a specified heteroatom species (N, O, and S) one or more of), which are monocyclic, bridged, or spirocyclic, and each ring is saturated. Bridged rings refer to polycyclic rings in which two or more atoms are shared between monocyclic rings. A spiro ring refers to a polycyclic ring in which a single atom is shared between the monocyclic rings. Heterocycloalkyl groups include, but are not limited to, azetidinyl, tetrahydropyrrolyl, tetrahydrofuranyl, morpholinyl, piperidinyl, and the like.

The term "pharmaceutically acceptable salt" refers to a salt of a compound obtained by reacting a compound with a pharmaceutically acceptable (relatively non-toxic, safe, patient-friendly) acid or base. When the compound contains a relatively acidic functional group, base addition salts can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to, sodium, potassium, calcium, aluminum, magnesium, bismuth, ammonium, and the like. When the compound contains relatively basic functional groups, acid addition salts can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride, sulfate, mesylate, and the like. See specifically Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, 2002).

The term "solvate" refers to a substance formed by crystallizing a compound with a solvent including, but not limited to, water, methanol, ethanol, and the like. Solvates are divided into stoichiometric and non-stoichiometric solvates.

The term "solvate of a pharmaceutically acceptable salt" refers to a compound with a pharmaceutically acceptable (relatively non-toxic, safe, suitable for patient use) acid or base, a solvent (including but not limited to: water, methanol, ethanol) etc.) are combined to form substances, wherein the pharmaceutically acceptable salt has the same meaning as the term "pharmaceutically acceptable salt" above and the solvent is stoichiometric or non-stoichiometric. Solvates of pharmaceutically acceptable salts include, but are not limited to, hydrochloride monohydrate.

The term "pharmaceutical excipients" refers to the excipients and additives used in the production of pharmaceuticals and the formulation of prescriptions, and are all substances contained in pharmaceutical preparations other than active ingredients. For details, please refer to the Pharmacopoeia of the People's Republic of China (2020 edition) or the Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009).

Unless otherwise defined, all technical and scientific terms used herein have the standard meaning in the art to which the claimed subject matter belongs. If more than one definition exists for a term, the definitions herein prevail.

It should be understood that singular forms such as "a" or "an" used in the present invention include plural referents unless stated otherwise. In addition, the term "comprising" is an open definition rather than a closed one, that is, it includes the contents specified in the present invention, but does not exclude other aspects.

Unless otherwise specified, the present invention adopts traditional methods of mass spectrometry and elemental analysis, and each step and condition may refer to the conventional operation steps and conditions in the art.

Unless otherwise indicated, the present invention employs standard nomenclature and standard laboratory procedures and techniques of analytical chemistry, synthetic organic chemistry, and optics. In some cases, standard techniques are used for chemical synthesis, chemical analysis, and performance testing of light-emitting devices.

In addition, it should be noted that, unless clearly stated otherwise, the description mode "...independently" used in the present invention should be understood in a broad sense, meaning that the described individuals are independent of each other and may be independent of each other. are the same or different specific groups. In more detail, the description mode "...independently" can either mean that in different groups, the specific options expressed between the same symbols do not affect each other; it can also mean that in the same group, the same symbols are The specific options expressed between them do not affect each other.

It can be understood by those skilled in the art that, according to the conventions used in the art, the formula used in the structural formula of the group described in this application

It means that the corresponding group is connected with other fragments and groups in the compound through this site.

On the basis of not violating common knowledge in the art, the above preferred conditions can be combined arbitrarily to obtain preferred examples of the present invention.

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

The positive improvement effect of the present invention is that the present invention provides a pyrazoloquinazoline compound, a preparation method and application thereof, and the compound has at least any one of the following advantages, which has obvious inhibitory effect on PLK1 and can further regulate the half-life of the drug.

Detailed ways

The present invention is further described below by way of examples, but the present invention is not limited to the scope of the described examples. The experimental methods that do not specify specific conditions in the following examples are selected according to conventional methods and conditions, or according to the product description.

Example 1

first step

Compound 1a (480 mg, 2.0 mmol) and 2-hydrazinoethanol (152 mg, 2.0 mmol) were dissolved in 5 mL of glacial acetic acid at room temperature and stirred for 5 hours. After the reaction was completed, the crude product was dispersed in ethyl acetate (20 mL) and water (30 mL) after removing most of the glacial acetic acid, and the aqueous phase was extracted with ethyl acetate (20 mL × 1), and the organic phase was successively dissolved in saturated sodium bicarbonate ( 20 mL x 1) and saturated brine (20 mL x 1), washed with anhydrous sodium sulfate, filtered, and concentrated to obtain compound 1b (430 mg, yield: 85%).

second step

Under nitrogen protection, a solution of compound 1b (800 mg, 3.2 mmol) and dimethylformamide di-tert-butyl acetal (709 mg, 3.5 mmol) in dimethylformamide (20 mL) was heated to 60 °C and reacted for 2 hours . The reaction ends. Cool, add water (50mL) to the reaction solution, extract with ethyl acetate (20mL×2), wash the organic phase with water (20mL×1) and saturated brine (20mL×1) successively, dry over anhydrous sodium sulfate, filter , concentrated to obtain compound 1c (750 mg, yield: 76%).

third step

Under nitrogen, a solution of compound 1c (750 mg, 2.4 mmol) and compound 1d (728 mg, 2.4 mmol) in dimethylformamide (10 mL) was heated to 120°C and stirred for 2 hours. The reaction ends. Cool, add 30 (mL) of water, extract with ethyl acetate (20 mL x 2), wash the organic phase with water (20 mL x 1) and saturated brine (20 mL x 1) successively, dry over anhydrous sodium sulfate, filter, and concentrate, The obtained crude product was subjected to silica gel column chromatography (methanol/dichloromethane=0/100%) to obtain compound 1e (780 mg, yield: 59%).

LCMS (M+H)+m/z: 542/544.

the fourth step

To a solution of compound 1e (780 mg, 1.4 mmol) in ethanol (10 mL) was added potassium hydroxide in ethanol (1.5 M, 3 mL) at room temperature. The resulting reaction solution was continuously stirred for 4 hours. The reaction ends. Filtration gave compound 1f (650 mg, yield: 82%).

the fifth step

To a solution of compound If (650 mg, 1.2 mmol), ammonium chloride (80 mg, 1.5 mmol) and triethylamine (310 mg, 3.6 mmol) in dimethylformamide (5 mL) was added 2-( 7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (684 mg, 1.8 mmol), and reacted for 2 hours. The reaction ends. The reaction solution was diluted with water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phase was washed successively with water (10 mL x 1) and saturated brine (10 mL x 1), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain Compound 1 g (510 mg, yield: 83%).

LCMS (M+H)+m/z: 513/515.

Step 6

Under nitrogen protection, compound 1g (51mg, 0.1mmol), 8-methyl-3,8-diazabicyclo[3.2.1]octane dihydrochloride (24mg, 0.12mmol), cesium carbonate ( 130 mg, 0.4 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (24 mg, 0.04 mmol) and tridibenzylideneacetone dipalladium (18 mg, 0.02 mmol) A solution of 1,4-dioxane (2 mL) was heated to reflux for 4 hours. The reaction ends. Cool, add water (10mL) to the reaction solution, extract with ethyl acetate (15mL×3), wash the organic phase with water (10mL×1) and saturated brine (10mL×1) successively, dry over anhydrous sodium sulfate, filter , concentrated, and the obtained crude product was purified by preparative HPLC to obtain compound 1 (1.5 mg, yield: 27%).

LCMS (M+H)+m/z: 559.

Example 2

first step

According to the method of Example 1, compound 2 (3.0 mg) was obtained from compound 1 g.

LCMS (M+H)+m/z: 559.

Example 3

first step

According to the method of Example 1, compound 3 (1.6 mg) was obtained from compound 1 g.

LCMS (M+H)+m/z: 545.

Example 4

first step

According to the method of Example 1, compound 4 (1.3 mg) was obtained from compound 1 g.

LCMS (M+H)+m/z: 545.

Example 5

first step

According to the method of Example 1, compound 5 (2.0 mg) was obtained from compound 1 g.

LCMS (M+H)+m/z: 545.

Example 6

first step

1,2-cyclohexanedione 6F_1 (100 g, 0.892 mol) was dissolved in a mixed solvent of toluene (700 mL) and MeOH (500 mL). p-Toluenesulfonic acid (15.4 g, 89.2 mmol) was added and the solution was stirred at 100°C for 48 hours. The solvent was evaporated and the residue was dissolved in DCM (500 mL) and washed with a saturated solution of _NaHCO3 (500 mL). The organic phase was dried _over _Na2SO4 and concentrated. Purification by silica gel column chromatography (0%-30% EA in PE) afforded 2-methoxycyclohexyl-2-en-1-one 6F_2 (42 g, 37.3%) as an oil. ESI-MS (M+H) ⁺ =127.

second step

LiHMDS (51 mL, 51.0 mmol, 1 M in THF) was added dropwise to 2-methoxycyclohexyl- ₂ -en-1-one (6.40 g, 45.7 mmol) at -50 °C under N atmosphere solution in THF (60 mL). After stirring at -50°C for 30 minutes, diethyl oxalate (7.47 g, 51.0 mmol) was added. The solution was stirred at 25°C for 16 hours. After the reaction, water (100 mL) was added, and 1N HCl was added to adjust the pH to 4-5, and the resulting solution was extracted with EA (50 mL×3). The organic layer was dried _over _Na2SO4 and evaporated to dryness. The crude product was purified by column (0%-15% EA in PE) to give 2-(3-methoxy-2-oxocyclohex-3-en-1-yl)-2-oxoacetic acid as a pale yellow oil Ethyl ester 6F_3 (8.2 g, 71.4%). ¹ HNMR (400MHz, CDCl ₃ ) δH 14.86(s, 1H), 5.89(t, 1H), 4.35(q, 2H), 3.66(s, 3H), 2.90(t, 2H), 2.45-2.38(m, 2H)), 1.38(t, 3H).

third step

To ethyl 2-(3-ethoxy-2-oxocyclohex-3-en-1-yl)-2-oxoacetate 6F_3 (3.0 g, 0.012 mmol) in ethanol (30 mL) at 25 °C To the solution was added hydroxyethylhydrazine (0.6 mL, 0.012 mmol). The reaction was stirred at 80°C for 5 hours. The solvent was then evaporated and the residue was redissolved with DCM (30 mL). The organic layer was washed with water, dried _over _Na2SO4 and concentrated. The residue was dissolved in THF (12 mL) and HCl (12 mL, 1 mol/L) and stirred at 25 °C for 2 h, the reaction was quenched with saturated NaHCO ₃ solution (50 mL), EA (50 mL×3), combined The organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to give 1-(2-hydroxyethyl)-7-oxo-4,5,6,7-tetrahydro as a brown oil -1H-Indazole-3-carboxylic acid ethyl ester 6F_3 (2.4 g, crude). ESI-MS (M+H) ⁺ =253.

the fourth step

6F_3 (4 g, 15.9 mmol) was dissolved in DMF (24 mL) at 25°C, then 8 mL of DMFDMA was added. The resulting mixture was stirred at 80 °C for 20 h, then cooled to 25 °C, concentrated to remove solvent, and the crude product was purified by column (20%-50% EA in PE) to give 6F (3.25 g, 66.7%) as a yellow solid. 1HNMR(600MHz,DMSOd6)δH 7.49(s,1H),4.83(t,1H),4.60(t,2H),4.26(q,2H),3.72(q,2H),3.12(s,6H),2.95 -2.80(m, 4H), 1.29(t, 3H).

the fifth step

5-Bromo-4-fluoro-2-methoxyaniline 6A (7.5 g, 34.3 mmol) was dissolved in EtOH (90 mL) and _Ac2O (8.7 g, 85.6 mmol) was added. The mixture was stirred at 25 °C for 2.5 h, and the solvent was evaporated to give N-(5-bromo-4-fluoro-2-methoxyphenyl)acetamide 6A_1 (8 g crude) as a brown solid. ESI-MS (M+H) ⁺ =262.

Step 6

In a round bottom flask was added N-(5-bromo-4-fluoro-2-methoxyphenyl)acetamide 6A_1 (1.5 g, 5.74 mmol), Davephos (90 mg, 0.223 mmol), Pd2(dba)3 ( 105 mg, 0.115 mmol) and THF (7.5 mL). The flask was evacuated and backfilled with nitrogen. LiHMDS (1 mol/L in tetrahydrofuran, 12.63 mL) and N-methylpiperazine (690 mg, 6.89 mmol) were added dropwise, and the reaction was refluxed at 70° C. for 1 h. The reaction mixture was added with saturated aqueous NH4Cl solution (10 mL), extracted with EA (10 mL×3), the organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure. The residue was purified by column chromatography (2.5%～5% methanol, dichloromethane) to obtain N-(4-fluoro-2-methoxy-5-(4-methylpiperazin-1-yl)phenyl)ethyl Amide 6A_2 (400 mg, 24.8%) was a pale yellow solid. LCMS(M+H)+=282.

Step 7

N-(4-Fluoro-2-methoxy-5-(4-methylpiperazin-1-yl)phenyl)acetamide 6A-2 (400 mg, 1.67 mmol) was dissolved in EtOH (4 mL), stirred at room temperature, To the solution was added HCl (1.2 mL, 12 mol/L). The mixture was stirred at 80° C. for 6 h. After the reaction was completed, 2 mol/L NaOH was added to adjust the pH of the reaction mixture to 8-9. The aqueous phase was extracted with EA (10 mL x 3), the organic layer was washed with brine, dried _over _Na2SO4 and concentrated. The residue was purified by column (1%-50% EA in PE) to give 4-fluoro-2-methoxy-5-(4-methylpiperazin-1-yl)aniline 6B (300 mg, 88.2%) as a yellow solid . ¹ HNMR (400MHz, DMSO-d6)δH 6.68(d,1H), 6.37(d,1H), 4.47(s,2H), 3.69(s,3H), 2.85(brs,4H), 2.43(brs,4H) ),2.20(s,3H).

Step 8

4-Fluoro-2-methoxy-5-(4-methylpiperazin-1-yl)aniline 6B (150 mg, 0.514 mmol) was dissolved in hydrochloric acid (6 mol/L, 1 mL), and cyanamide (345 mg) was added. , 4.11 mmol, 50% in _H2O ) and the reaction was stirred at 80 °C for 48H. The mixture was cooled to 25°C, diluted with water (3 mL), and extracted with dichloromethane. The pH of the aqueous phase was adjusted to >11 by adding 2 mol/L NaOH. The aqueous phase was extracted with EA (10 mL x 3), dried over Na ₂ SO ₄ and concentrated to give 1-(4-fluoro-2-methoxy-5-(4-methylpiperazine-1) as a brown solid -yl)phenyl)guanidine 6C (150 mg, crude).

Step 9

1-(4-Fluoro-2-methoxy-5-(4-methylpiperazin-1-yl)phenyl)guanidine 6C (300 mg, 0.98 mmol) and 6-((dimethylamino) at 25°C )methylene)-1-(2-hydroxyethyl)-7-oxo-4,5,6,7-tetrahydro-1H-indazole-3-carboxylic acid ethyl ester 6F in DMF (3.5 mL) stirring in. After stirring at 80°C for 12 hours, the reaction mixture was evaporated under reduced pressure. The residue was purified by column (2%-5% MeOH in DCM) to give ethyl 8-((4-fluoro-2-methoxy-5-(4-methylpiperazin-1-yl)phenyl) )amino)-1-(2-hydroxyethyl)-4,5-dihydro-1H-pyrazolo[4,3-h]quinoxaline-3-carboxylic acid ethyl ester 6D (150 mg, 26.7%), For light yellow solid. LCMS(M+H)+=526.

Step 10

Ethyl 8-((4-fluoro-2-methoxy-5-(4-methylpiperazin-1-yl)phenyl)amino)-1-(2-hydroxyethyl)-4,5 - Dihydro-1H-pyrazolo[4,3-h]quinoxaline- ₃ -carboxylic acid ethyl ester 6D (110 mg, 0.222 mmol) in NH in MeOH (1.5 mL, 7 M) at 80 °C under stirring in a 10 mL sealed tube for 36 hours. After the reaction was completed, the reaction mixture was evaporated under reduced pressure, and the residue was purified by column chromatography (DCM:MeOH=10:1) to give 8-((4-fluoro-2-methoxy-5 as a pale yellow solid) -(4-Methylpiperazin-1-yl)phenyl)amino)-1-(2-hydroxyethyl)-4,5-dihydro-1H-pyrazolo[4,3-h]quinazole Line-3-carboxamide 6 (12.0 mg, 11.5%).

¹ HNMR (400MHz, CD ₃ OD) δH 8.27(s, 1H), 7.70(d, 1H), 6.88(d, 1H), 4.74(s, 2H), 3.87(t, 2H), 3.84(s, 3H) ), 3.13-3.03 (m, 6H), 2.91-2.86 (m, 2H), 2.65 (brs, 4H), 2.36 (s, 3H). LCMS(M+H)+=497.2.

Example 7

first step

Under N atmosphere, put (Z)-6-((dimethylamino)methylene)-1-( ₂ -hydroxyethyl)-7-oxo-4,5,6,7-tetrahydro- To a solution of 1H-indazole-3-carboxylic acid ethyl ester 6F (570 mg, 1.88 mmol) in DMF (50 ml) was added guanidine carbonate (849.4 mg, 4.64 mmol). After stirring at 110°C for 16 hours, the reaction mixture was added to water (50 mL), filtered and dried in vacuo. The residue was purified by column chromatography (20%-50% EA/PE) to give 8-amino-1-(2-hydroxyethyl)-4,5-dihydro-1H-pyrazolo as pale yellow solid Ethyl [4,3-h]quinazoline-3-carboxylate 7F_1 (307 mg, 53.8%). ESI-MS (M+H) ⁺ =304.

second step

To 8-amino-1-( ₂ -hydroxyethyl)-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxylic acid ethyl ester 7F-1( To a solution of 303 mg, 1.00 mmol) in THF (30 ml) was added cuprous iodide (57.1 mg, 0.300 mmol), iodine (126.5 mg, 0.500 mmol), cesium iodide (259 mg, 1.00) and isoamyl nitrite (176 mg) , 1.50 mmol). The reaction was stirred at 80°C for 12 hours. The reaction mixture was diluted with DCM (50 mL), washed with 20% NH ₃ ·H ₂ O (80 mL), saturated aqueous Na ₂ S ₂ O ₃ (80 mL), brine (80 mL), dried over anhydrous sodium sulfate, filtered, reduced pressure concentrate. The residue was purified by column chromatography (2.5%-10% MeOH in DCM) to give 1-(2-hydroxyethyl)-8-iodo-4,5-dihydro-1H-pyrazole as a yellow solid and [4,3-h]quinazoline-3-carboxylic acid ethyl ester 7F_2 (300 mg, 52%). ESI-MS (M+H) ⁺ =415.

third step

A round bottom flask was charged with 1-bromo-2-fluoro-4-(trifluoromethoxy)benzene 7A (27 g, 104 mmol), and potassium nitrate (14 g, 138 mmol), concentrated sulfuric acid (60 ml) were added under _N2 ). The reaction was stirred at 25°C for 2 hours. The reaction mixture was quenched with water (200 mL), extracted with DCM (50 mL x 3), the combined organic layers were washed with saturated aqueous NaHCO ₃ (100 mL), brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. The residue was purified by column (0%-5% EA in PE) to give 1-bromo-2-fluoro-5-nitro-4-(trifluoromethoxy)benzene 7B as a pale yellow oil (30 g, 95%). ESI-MS (M+H) ⁺ =303.

the fourth step

To a solution of 1-bromo- ₂ -fluoro-5-nitro-4-(trifluoromethoxy)benzene 7B (25.0 g, 12.1 mmol) in AcOH (50 mL) and EtOH (50 mL) under N2 Iron powder (22.0 g, 0.393 mol) was added to it. The reaction was stirred at 25°C for 3 hours. The reaction mixture was concentrated and diluted with water (100 ml), extracted with EA (30 mL x 3), the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure. The residue was purified by column (5%-20% EA in PE) to give 5-bromo-4-fluoro-2-(trifluoromethoxy)aniline 7C (21.2 g, 93.8%) as a yellow solid. ¹ H NMR (400 MHz, CDCl3) δH 7.00-6.95 (m, 2H), 3.79 (brs, 2H). ESI-MS (M+H) ⁺ = 274.1.

the fifth step

According to the synthetic method of Example 6, starting from compound 7C, compound 7D is obtained through two-step reaction.

Step 6

To 1-( ₂ -hydroxyethyl)-8-iodo-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxylic acid ethyl ester 7F_2( To a solution of 126.6 mg, 0.432 mmol) in DMF (6 ml) was added 4-fluoro-5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)aniline 7D (60 mg, 0.144 mmol), Pd(OAc) ₂ (3.2 mg, 0.0144 mmol), BINAP (18.0 mg, 0.0289 mmol), K2CO3 ₍ 59.6 _mg , 0.432 mmol). The reaction was stirred at 80°C for 4 hours. The reaction mixture was concentrated and diluted with water (20 mL), extracted with EA (10 mL x 3), the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure. The residue was purified by column chromatography (10% MeOH in DCM) to give 8-((4-fluoro-5-(4-methylpiperazin-1-yl)-2-(tris) as a pale yellow solid Fluoromethoxy)phenyl)amino)-1-(2-hydroxyethyl)-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxylate ethyl ester 7F (75 mg, 16%). ESI-MS (M+H) ⁺ =580.

Step 7

According to the synthetic method of Example 6, compound 7 (5.2 mg, 7.3%) was obtained by one-step reaction from compound 7F (75 mg, 0.129 mmol).

¹ HNMR (400MHz, CD ₃ OD) δH 8.27(s, 1H), 7.45(d, 1H), 7.17(d, 1H), 4.78-4.74(m, 2H), 3.81(t, 2H), 3.26-3.21 (m, 4H), 3.10-3.05 (m, 2H), 2.90-2.83 (m, 2H), 2.67-2.62 (m, 4H), 2.36 (s, 3H).

LCMS(M+H)+m/z=551.2.

Example 8

first step

According to the synthetic method of Example 6, starting from compound 8-1, compound 8A is obtained through 3 steps. ¹ HNMR (400MHz, DMSO d6)δH 8.15(s,1H), 7.95(d,1H), 7.26-7.15(m,9H), 7.05(d,6H), 6.87(d,1H), 4.23(q, 2H), 3.80(s, 3H), 3.00(t, 2H), 2.88-2.81(m, 4H), 2.67-2.58(m, 2H), 2.53-2.35(m, 7H), 1.24(t, 3H) .

second step

To 8-((4-fluoro-5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl)amino)-1-(2-hydroxyethyl) at 25°C )-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxylic acid ethyl ester 8A (100 mg, 0.138 mmol) in DCM (5 mL) was added TFA (0.5 mL) , and stirred at 25 °C for 4 hours. The reaction was quenched by adding saturated aqueous NaHCO ( ₃₀ mL), extracted with DCM (15 mL×2), the organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered, evaporated under reduced pressure, and the residue was subjected to column chromatography (2%～ 8% MeOH in DCM) to give 8-((4-fluoro-2-methoxy-5-(4-methylpiperazin-1-yl)phenyl)amino)-4,5-dihydro -1H-Pyrazolo[4,3-h]quinazoline-3-carboxylic acid ethyl ester 8B (40 mg, 60.2%). ESI-MS (M+H) ⁺ =482.

third step

According to the synthetic method of Example 6, compound 8C is obtained from compound 8B through one-step reaction.

third step

To 8-((4-fluoro-5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl)amino)-1-(2-hydroxyl at 0°C Ethyl)-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxamide 8C (30 mg, 0.066 mmol) in DMF ( ₃ mL) was added _Cs2CO3 (32.6 mg, 0.1 mmol) and 2-chloroacetonitrile (6.0 mg in 1 mL DMF, 0.08 mmol). After stirring at 25°C for 4 hours, the reaction mixture was quenched with water (10 mL). The mixture was extracted with EA (15 mL x 2), the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure. The residue was purified by column chromatography (DCM:MeOH=15:1) to give 1-(cyanomethyl)-8-((4-fluoro-2-methoxy-5-(4) as a pale yellow solid -Methylpiperazin-1-yl)phenyl)amino)-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxamide 8 (16.7 mg, 51.2% ).

¹ HNMR (400MHz, CD ₃ OD) δH 8.31(s, 1H), 7.68(d, 1H), 6.88(d, 1H), 5.88(s, 2H), 3.84(s, 3H), 3.15-3.05(m , 6H)), 2.90(t, 2H), 2.63-2.58(m, 4H), 2.33(s, 3H).

LCMS[M+H]+: 492.3.

Example 9

According to the synthetic method of Example 8, compound 9 was obtained from compound 8-3 and compound 7D through 4-step reaction.

¹ HNMR (400MHz, CD ₃ OD) δH 8.30(s, 1H), 7.45(d, 1H), 7.14(d, 1H), 5.80(s, 2H), 3.18-3.05(m, 6H), 2.90(t ,2H),2.68-2.60(m,4H),2.36(s,3H).

LCMS[M+H]+: 446.3.

Example 10

According to the synthetic methods of Examples 7 and 8, compound 10 was obtained from compound 8-3 and compound 10-1 through 4-step reaction.

¹ HNMR (400MHz, CD ₃ OD) δH 8.37(s, 1H), 7.54(s, 1H), 7.26(d, 1H), 6.85(d, 1H), 5.87(s, 2H), 3.31-3.25(m , 4H), 3.14 (t, 2H), 2.96 (t, 2H), 2.71-2.63 (m, 4H), 2.38 (s, 3H).

LCMS[M+H]+: 528.2.

Example 11

According to the synthetic method of embodiment 9, starting from compound 10C, compound 11 is obtained through 1-step reaction.

¹ HNMR (400MHz, CD ₃ OD) δH 8.31(s, 1H), 7.45(s, 1H), 7.25-7.20(m, 1H), 6.85-6.77(m, 1H), 5.10-5.02(m, 2H) ,4.62-4.51(m,2H),4.34-4.28(m,1H),3.26(brs,4H),3.15-3.08(m,2H),2.94-2.86(m,2H),2.61(brs,4H) , 2.55-2.49(m, 1H), 2.36(s, 3H), 2.35-2.29(m, 1H).

LCMS[M+H]+: 559.2.

Example 12

first step

Methyl 1-hydroxycyclopropane-1-carboxylate 12A (3.0 g, 25.8 mmol) was dissolved in MeOH (15 mL) and hydrazine hydrate (6.37 g, 258 mmol) was added. The mixture was stirred at 60°C for 16 hours and the solvent was evaporated. The residue was triturated with EA (30 mL) for 0.5 h and filtered to give 1-hydroxycyclopropane-1-carboxyhydrazide 12E_1 (2.8 g, 93.3%) as a white solid. LCMS(M+H)+=117.

second step

A suspension of 1-hydroxycyclopropane- ₁ -carboxyhydrazide 12E_1 (3.0 g, 258 mmol) in THF (15 mL) was added dropwise BH ₃ ·THF (258 mL, 258 mmol, 1M). The reaction mixture was stirred at 60°C for 21 hours. MeOH (50 mL) was added dropwise to the reaction mixture at -5-0 °C under _N2 protection, then stirred at 20-25 °C for 0.5 h, and the reaction mixture was concentrated to give a residue. The residue was dissolved in 50 mL of MeOH and stirred at 80°C for 3 hours. The mixture was cooled to 25°C and evaporated to give a residue. The residue was triturated with DCM (30 mL) for 0.5 h and filtered. The filtrate was evaporated to give crude 1-(hydrazinomethyl)cyclopropan-1-ol 12E_2 as a pale yellow oil (2.2 g, 75%). ¹ H NMR (400 MHz, CDCl ₃ ) d 2.91 (s, 2H), 0.83 (t, J=6.0 Hz, 2H), 0.55-0.50 (m, 2H).

third step

According to the synthetic method of Example 9, compound 12 was obtained from compound 12E_2 and compound 6F_3 through 4-step reaction.

¹ HNMR (400MHz, CD ₃ OD) δH 8.28(s, 1H), 7.44(s, 1H), 7.20(d, 1H), 6.79(d, 1H), 4.74(s, 2H), 3.30-3.24(m ,4H),3.12-3.05(m,2H),2.90-2.84(m,2H),2.63-2.58(m,4H),2.33(s,3H),0.59-0.54(m,2H),0.49-0.46 (m, 2H).

LCMS[M+H]+: 559.2.

Example 13

According to the synthetic method of Example 9, starting from compound 10C, compound 13 was obtained through one-step reaction.

¹ H NMR (400MHz, DMSO-d6) δ 9.12 (s, 1H), 8.38 (s, 1H), 7.49–7.21 (m, 5H), 6.92 (dd, J=9.1, 3.0Hz, 1H), 4.79 (d, J=7.0Hz, 2H), 4.40 (t, J=7.0Hz, 2H), 4.31 (t, J=6.2Hz, 2H), 3.56 (d, J=12.3Hz, 3H), 3.20 (ddt , J=26.3, 15.4, 7.4Hz, 4H), 3.00 (t, J=7.5Hz, 4H), 2.90 (s, 4H), 2.83 (t, J=7.7Hz, 3H).

Example 14

According to the synthetic method of Example 9, starting from compound 10C, compound 14 was obtained through one-step reaction.

¹ H NMR (400MHz, CD ₃ OD-d4) δppm 8.51 (s, 1H), 7.53 (d, J=2.9Hz, 1H), 7.36-7.21 (m, 1H), 6.87 (dd, J=9.1, 3.0 Hz,1H),4.76(d,J＝12.7Hz,2H),4.50(d,J＝13.2Hz,1H),4.42(d,J＝12.7Hz,1H),3.65(d,J＝2.0Hz, 2H), 3.26(dt, J＝18.0, 6.2Hz, 6H), 3.06(t, J＝7.7Hz, 2H), 2.67(t, J＝5.0Hz, 4H), 2.40(s, 3H), 1.42( s, 3H).

LCMS[M+H]+: 573.2.

Example 15

According to the synthetic method of Example 9, starting from compound 10C, compound 15 was obtained through one-step reaction.

¹ HNMR (400MHz, CD ₃ OD) δH 8.35(s, 1H), 7.53(s, 1H), 7.26(d, 1H), 6.85(d, 1H), 5.00-4.88(m, 1H), 4.65-4.58 (m,1H),3.29(s,4H),3.15-3.08(m,2H),2.93(t,2H),2.80(s,4H),2.49(s,3H),2.21-2.12(m,1H) ), 1.39-1.32 (m, 2H).

LCMS[M+H]+: 579.2.

Example 16

first step

To a solution of 16A (193 mg, 1 mmol) in DMF (5 mL) at 0 °C was added _Cs2CO3 (650 mg, ₂ mmol) and (2-bromoethoxy)tert-butyldimethylsilane (480 mg, 2 mmol) . Stir overnight at 25°C and quench the reaction mixture with water (10 mL). The mixture was extracted with EA (15 mL x 2), the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure. The residue was purified by column chromatography (PE:EA=5:1) to give 16B (220 mg, 64%) as a white solid.

second step

16B 510 mg was treated with tris(dimethylamino)methane (2 mL) and the reaction was stirred at 90 °C overnight, the volatiles were removed under reduced pressure, and the residue was used without further purification.

third step

According to the synthetic method of Example 10, compound 16D is obtained by one-step reaction from compound 16B and compound 15-1.

the fourth step

Methyl 9-(2-((tert-butyldimethylsilyl)oxy)ethyl)-2-((5-(4-methylpiperazin-1-yl)-2-(tri Fluoromethoxy)phenyl)amino)-6,9-dihydro-5H-pyrrolo[3,2-h]quinazoline-7-carboxylic acid methyl ester 16D (330 mg, 0.5 mmol) was suspended in dioxygen hexacyclic ring (5 mL) and treated with 2N NaOH solution (5 mL, 10 mmol) at reflux temperature overnight, _H2O (50 mL) was added and the solution was acidified to pH=4 with 2N HCl. Concentrated under reduced pressure, the residue was subjected to column chromatography (DCM:MeOH 10:1) to give the product 9-(2-hydroxyethyl)-2-((5-(4-methylpiperazin-1-yl) as a yellow solid) -2-(Trifluoromethoxy)phenyl)amino)-6,9-dihydro-5H-pyrrolo[3,2-h]quinazoline-7-carboxylic acid (240 mg, 90% yield). LC-MS (M+H)+: 533.

the fifth step

9-(2-hydroxyethyl)-2-((5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl)amino)-6,9-di Hydrogen-5H-pyrrolo[3,2-h]quinazoline-7-carboxylic acid (240 mg, 0.45 mmol) was dissolved in DMF, to the reaction mixture were added HOBt (121 mg, 0.9 mmol) and EDCI (172 mg, 0.9 mmol) ), stirred for 0.5h. Subsequently, _NH4OH (4eq) was added. The reaction mixture was stirred at room temperature for 14 hours, the reaction mixture was concentrated under reduced pressure to give 9-(2-hydroxyethyl)-2-((5-(4-methylpiperazin-1-yl) by preparative HPLC -2-(Trifluoromethoxy)phenyl)amino)-6,9-dihydro-5H-pyrrolo[3,2-h]quinazoline-7-carboxamide, white solid (106 mg, yield 47%).

¹ H NMR(400MHz,DMSO-d6)δ8.52(s,1H),8.14(s,1H),7.55(s,1H),7.24(d,J=3.0Hz,1H),7.21(dq,J ＝9.1,1.4Hz,1H),6.79(d,J＝3.0Hz,1H),6.77(d,J＝3.0Hz,1H),4.71(t,J＝5.3Hz,1H),4.35(t,J =5.0Hz,2H),3.46-3.40(m,3H),3.15(dd,J=6.4,3.7Hz,4H),2.98(t,J=7.7Hz,2H),2.72(t,J=7.7Hz , 2H), 2.46 (t, J=5.0Hz, 4H), 2.24 (s, 3H).

Example 17

first step

To a solution of cyclopropylmethanol 17A (721.00 μL, 9.09 mmol) in THF (24 mL) was added dropwise LiHMDS (1 M in THF, 6.06 mL, 9.09 mmol) at 5°C. After 30 minutes, 4-fluoro-3-nitrobromobenzene (1.11 mL, 9.09 mmol) was added rapidly and the reaction mixture was allowed to warm to room temperature and stirred overnight. The reaction mixture was quenched with water and extracted with EA, the collected organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by silica gel column chromatography (PE/EA, 100:0 to 20:1) to give 3.02 g of 4-bromo-1-(cyclopropylmethoxy)-2-nitrobenzene 17B. LCMS (M+H) ⁺ =272.

second step

Under N atmosphere, to dry 1,4 _- dioxane (20 mL) was added 4-bromo-1-(cyclopropylmethoxy)-2-nitrobenzene (587 mg, 2.16 mmol), XANTPHOS ( 0.37 g, 0.65 mmol), Pd2(dba ₎₃ ₍ 0.39 g, 0.43 mmol, Aldrich), _Cs2CO3 (1.4 g, _4.3 mmol) and 1-methylpiperazine (0.43 g, 4.3 mmol). After heating at 90 °C overnight, the reaction was quenched with water, extracted with ethyl acetate (50 mL x 3), the organic phase was dried over sodium sulfate, concentrated, and purified by column chromatography (dichloromethane/methanol 20:1), 1-(4-(Cyclopropylmethoxy)-3-nitrophenyl)-4-methylpiperazine 17C (383 mg, 1.32 mmol, 61%) was obtained as a tan solid. ESI-MS(M+H)+=292.

third step

1-(4-(Cyclopropylmethoxy)-3-nitrophenyl)-4-methylpiperazine 17C (291 mg, 1 mmol) was dissolved in a mixed solvent of acetic acid (10 mL) and ethanol (10 mL) , added zinc powder (650 mg, 10 mmol), stirred at 80° C. for 3 h, filtered through celite, concentrated, and purified by column chromatography (dichloromethane/methanol 20:1) to obtain 2-(cyclopropylmethoxy) )-5-(4-methylpiperazin-1-yl)aniline 17D (185 mg, 0.71 mmol, 71%). ESI-MS(M+H)+=262.

the fourth step

According to the synthetic method of Example 7, compound 17, 2-((2-(cyclopropylmethoxy)-5-(4-methylpiperazin-1-yl) was obtained from compound 17D and 7F_2 through 2-step reaction )phenyl)amino)-9-(2-hydroxyethyl)-6,9-dihydro-5H-pyrrolo[3,2-h]quinazoline-7-carboxamide.

1H NMR(400MHz,DMSO-d6)δ8.43(s,1H),8.05(s,1H),7.74(d,J=2.8Hz,1H),7.50(s,1H),7.29(s,1H) ,6.94(d,J＝8.9Hz,1H),6.60(dd,J＝8.8,2.9Hz,1H),4.78(t,J＝5.6Hz,2H),3.85(d,J＝6.9Hz,2H) ,3.80(t,J＝5.6Hz,2H),3.05(t,J＝5.0Hz,4H),3.00(d,J＝7.7Hz,2H),2.83(t,J＝7.6Hz,2H),2.49 (t, J=4.9 Hz, 4H), 2.25 (s, 3H), 1.29–1.18 (m, 1H), 0.60–0.51 (m, 2H), 0.36–0.28 (m, 2H).

LCMS[M+H]+: 519.3.

Example 18

18A was prepared by the method described in patent CN101563351B. A stirred solution of 18A (100 mg, 0.178 mmol) in THF (2 mL) was added to LiAlH4 (28 mg, 0.712 mmol) in a 50 mL sealed tube at -78 °C. After stirring at -78°C for 24 hours, the reaction mixture was cooled to 0°C and water (20 mL) was added. The mixture was extracted with EA (20 mL×3), the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure. The residue was purified by column chromatography (2.5%-10% MeOH in DCM) to give compound 18 (15 mg, 16.2%) as a pale yellow solid.

1HNMR(400MHz,CD3OD)δH 8.27(s,1H),7.54(d,1H),7.20(d,1H),6.79(d,1H),4.71(t,2H),4.60(s,2H),3.79 (t,2H),3.28-3.21(m,4H),2.90-2.80(m,4H),2.66(m,4H),2.38(s,3H).

LC-MS [M+H]+: 520.20.

Example 19

A stirred solution of 18A (0.15 g, 0.267 mmol) in DCM (6 mL) at 25 °C in a 50 mL sealed tube, EDCI (70.5 mg, 0.347 mmol), DMAP (16.3 mg, 0.134 mmol), O-methyl- Hydroxylamine chloride salt (35.3 mg, 0.4 mmol). After stirring at 30°C for 2 hours, after concentration to remove the solvent, the crude product was purified by column chromatography (DCM:MeOH=10:1) to give 19 (30 mg). ¹ HNMR (400MHz, CD ₃ OD) δ _H 8.30(s, 1H), 7.51(d, 1H), 7.21(d, 1H), 6.79(d, 1H), 4.76(t, 2H), 3.82(t, 2H), 3.79(s, 3H), 3.30-3.22(m, 4H), 3.07(t, 2H), 2.88(t, 2H), 2.64-2.61(m, 4H), 2.35(s, 3H).

LC-MS [M+H]+: 563.30.

Example 20

A round bottom flask was charged with 17A (5 g, 0.023 mol), cyclopropanol (1.32 g, 0.023 mol), K2CO3 (9.42 _g , 0.068 mol) and DMF ( ₁₀ mL). The reaction was stirred at 25°C overnight. The reaction mixture was quenched by adding _H2O (100 mL), extracted with EA (100 mL x 3), the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. The residue was passed through a column (in PE

EA) Purification to give 2OA (5.3 g, 90.4%) as a yellow solid.

According to the synthetic method of Example 17, starting from compound 20A, compound 14 was obtained through 4-step reaction.

¹ HNMR(400MHz, CD3OD)δH 8.30(s,1H), 7.82(d,1H), 7.24(d,1H), 6.72(dd,1H), 4.90-4.88(m,2H), 3.94(t,2H) ),3.85-3.82(m,1H),3.15-3.12(m,4H),3.07(t,2H),2.87(t,2H),2.71-2.66(m,4H),2.39(s,3H), 0.79-0.77(m,2H),0.74-0.72(m,2H).

LC-MS[M+H]+: 505.30.

Example 21

A. PLK1 Kinase Activity Assay

Experimental Materials:

PLK1 Active was purchased from CARNA;

Casein Protein was purchased from SignalChem;

ADP-Glo Kinase Assay was purchased from Promega;

Kinase assay buffer III was purchased from SignalChem;

Nivo Multilabel Analyzer (PerkinElmer).

experimental method:

Dilute enzyme, substrate, ATP and inhibitor with kinase buffer from the kit. The compounds to be tested were diluted with 100% DMSO to 1 mM as the first concentration, and then 5-fold diluted to the eighth concentration with a drain gun, ie, from 1 mM to 0.013 μM. Dilute each concentration point of the compound by 20 times with 1X kinase buffer, prepare a compound working solution containing 5% DMSO, add 1 μL of each concentration gradient working solution of the compound to the microplate, and set up double wells. Add 2μl of PLK1 enzyme (15ng), 2μl of the mixture of substrate and ATP (20μM ATP, 0.2μg/μl Casein protein) to the microtiter plate, at this time the final compound concentration gradient is 10μM and diluted to 0.13nM, the reaction system is placed at 25 reaction for 60 minutes. After the reaction, 5 μl of ADP-Glo reagent was added to each well, and the reaction was continued at 25 degrees for 40 minutes. After the reaction was completed, 10 μL of kinase detection reagent was added to each well. After 30 minutes of reaction at 25 degrees, the PerkinElmer Nivo multi-label analyzer was used to read the chemiluminescence, and the integral Time 0.5 seconds.

data analysis:

Using the equation (Sample-Min)/(Max-Min)*100% to convert the raw data into inhibition rate, the IC ₅₀ value can be obtained by curve fitting with four parameters (log(inhibitor) vs.response in GraphPad Prism --Variable slope mode).

The following table shows the PLK1 kinase activity test results of the compounds of the present application.

化合物编号Compound number	PLK1 IC ₅₀(nM) PLK1 IC ₅₀ (nM)
66	47.1747.17
77	101.8101.8

88	824.9824.9
99	526526
1010	9.559.55
1111	8.578.57
1212	9.679.67
1313	18.9418.94
1414	50.1250.12
1515	14.6214.62
1616	6.396.39
1717	9.719.71

B. In Vitro Cell Proliferation Assay

The inhibitory activity of compounds on tumor cell proliferation was determined using the MTT assay. The HT-29 tumor cells in the logarithmic growth phase were inoculated into the culture plate according to a certain amount of cells, incubated for ₂₄ hours, and added with different concentrations of inhibitors. Add 20 μL of MTT solution to continue culturing for 4 h, dissolve the crystals with DMSO, and measure the OD value at a wavelength of 570 nm with an enzyme-linked immunosorbent assay to calculate IC ₅₀ .

The following table shows the test results of the HT-29 tumor cell proliferation inhibitory activity of the compounds of the present application.

化合物编号Compound number	HT-29 IC ₅₀(nM) HT-29 IC ₅₀ (nM)
66	269269
1010	111111
1111	232.2232.2
1212	7575
1515	7171

Claims

A pyrazoloquinazoline compound as shown in formula I, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof:

Wherein, R 1 is C 1 -C 4 alkoxy, C 1 -C 4 alkoxy substituted by one or more R 1-1 or 3-6 membered cycloalkyloxy; R 1-1 is independently It is halogen or 3-6 membered cycloalkane;

R 2 is H, halogen or C 1 -C 4 alkyl;

R 3 is a 5- to 10-membered heterocycloalkyl group or a 5- to 10-membered heterocycloalkyl group substituted by one or more R 3-1 ; in the 5- to 10-membered heterocycloalkyl group, the heteroatom is selected from N One or more of , O and S, the number of heteroatoms is 1-2; R 3-1 is independently C 1 -C 4 alkyl;

R 4 is cyano, 3-6 membered cycloalkane, 3-6 membered cycloalkane substituted by one or more R 4-1 , 3-6 membered heterocycloalkane or substituted by one or more R 4-2 3-6-membered heterocycloalkane; C 1 -C 4 alkyl substituted by one or more R 4-3 ; in the 3-6 membered heterocycloalkyl, the heteroatom is selected from N, O and S One or more of , the number of heteroatoms is 1-2; R 4-1 is independently halogen, hydroxyl or C 1 -C 4 alkyl; R 4-2 is independently halogen or C 1 -C 4 alkyl; R 4-3 is independently halogen, cyano or hydroxyl;

R 5 is H, hydroxy, C 1 -C 4 alkyl or C 1 -C 4 alkoxy.
The pyrazoloquinazoline compound represented by formula I, its pharmaceutically acceptable salt, its solvate or its solvate of pharmaceutically acceptable salt as claimed in claim 1, characterized in that , R 1-1 is independently F, Cl, Br or I, such as F;

And/or, R 1-1 is independently a 3- to 6-membered cycloalkane, and the 3- to 6-membered cycloalkane is cyclopropane, cyclobutane or cyclopentane, such as cyclopropane or cyclobutane;

And/or, when R 1 is C 1 -C 4 alkoxy, the C 1 -C 4 alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, such as methyl Oxygen;

And/or, when R 1 is C 1 -C 4 alkoxy substituted by one or more R 1-1 , the C 1 -C 4 alkoxy is methoxy, ethoxy, iso- Propoxy or tert-butoxy, such as methoxy;

And/or, when R 1 is a 3- to 6-membered cycloalkyloxy group, the 3- to 6-membered cycloalkoxy group is cyclopropyloxy, cyclobutyloxy or cyclopentyloxy, for example cyclopropyloxy or cyclobutyloxy;

And/or, when R 2 is halogen, the halogen is F, Cl, Br or I;

And/or, when R 2 is a C 1 -C 4 alkyl group, the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl such as methyl;

And/or, when R 3-1 is a C 1 -C 4 alkyl group, the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl radical or tert-butyl, such as methyl;

And/or, when R 3 is a 5- to 10-membered heterocycloalkyl substituted by one or more R 3-1 , the 5- to 10-membered heterocycloalkyl is a 6- to 9-membered heterocycloalkyl, For example, a 6- to 9-membered heterocycloalkyl group containing two Ns, such as piperazinyl, hexahydropyridazinyl, hexahydropyrimidinyl, 3,8-diazabicyclo[3.2.1]octyl, octanyl Hydropyrrole[1,2-a]pyrazinyl, 2,6-diazaspiro[3.4]octyl, 3,6-diazabicyclo[3.2.0]heptyl, 1,6-bis azaspiro[3.4]octyl or octahydropyrrole[3,4-c]pyrrolyl;

And/or, when R 4-1 is halogen, the halogen is F, Cl, Br or I, such as F;

And/or, when R 4-1 is a C 1 -C 4 alkyl group, the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl radical or tert-butyl, such as methyl or ethyl;

And/or, when R 4-2 is a C 1 -C 4 alkyl group, the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl radical or tert-butyl, such as methyl or ethyl;

And/or, when R 4-2 is halogen, the halogen is F, Cl, Br or I, such as F;

And/or, when R 4-3 is halogen, the halogen is F, Cl, Br or I, such as F;

And/or, when R 4 is 3-6 membered cycloalkane, the 3-6 membered cycloalkane is cyclopropane, cyclobutane or cyclopentane, such as cyclopropane or cyclobutane;

And/or, when R 4 is a 3- to 6-membered cycloalkane substituted by one or more R 4-1 , the 3- to 6-membered cycloalkane is cyclopropane, cyclobutane or cyclopentane, such as cycloalkane Propane or cyclobutane;

And/or, when R 4 is a 3-6 membered heterocycloalkane substituted by one or more R 4-2 , the 3-6 membered heterocycloalkane is a 4-membered heterocycloalkane containing one N or O or a 5-membered heterocycloalkane containing one N or O;

And/or, when R 5 is a C 1 -C 4 alkyl group, the C 1 -C 4 alkyl group is methyl, ethyl, isopropyl or tert-butyl, such as methyl or ethyl;

And/or, when R 5 is C 1 -C 4 alkoxy, the C 1 -C 4 alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, such as methyl Oxygen.
The pyrazoloquinazoline compound represented by formula I, its pharmaceutically acceptable salt, its solvate or its solvate of pharmaceutically acceptable salt as claimed in claim 1, characterized in that , the R 1 is

E.g

and/or, R3 is

and/or, R3 is

and/or, R3 is

And/or, when R 4 is a 3-6 membered cycloalkane substituted by one or more R 4-1 , the 3-6 membered cycloalkane substituted by one or more R 4-1 is:

And/or, when R 4 is a 3- to 6-membered heterocycloalkane, the 3- to 6-membered heterocycloalkane is

And/or, when R 4 is a 3-6 membered heterocycloalkane substituted by one or more R 4-2 , the 3-6 membered heterocycloalkane substituted by one or more R 4-2 is

And/or, when R 4 is C 1 -C 4 alkyl substituted by one or more R 4-3 , the C 1 -C 4 alkyl substituted by one or more R 4-3 is
The pyrazoloquinazoline compound represented by formula I, its pharmaceutically acceptable salt, its solvate or its solvate of pharmaceutically acceptable salt as claimed in claim 1, characterized in that , R 4 is a cyano group, a 3- to 6-membered cycloalkane substituted by one or more R 4-1 , a 3- to 6-membered heterocycloalkane, a 3- to 6-membered heterocycle substituted by one or more R 4-2 Alkanes, or C 1 -C 4 alkyl substituted by one or more R 4-3 ;

And/or, R 4-1 is halogen or hydroxyl;

And/or, R 4-2 is C 1 -C 4 alkyl;

And/or, R 4-3 is hydroxyl;

And/or, R 5 is H, hydroxy or C 1 -C 4 alkoxy.
The pyrazoloquinazoline compound represented by formula I, its pharmaceutically acceptable salt, its solvate or its solvate of pharmaceutically acceptable salt as claimed in claim 1, characterized in that , R 4 is a cyano group, a 3- to 6-membered cycloalkane substituted by one or more R 4-1 , a 3- to 6-membered heterocycloalkane, a 3- to 6-membered heterocycle substituted by one or more R 4-2 Alkanes, or C 1 -C 4 alkyl substituted by one or more R 4-3 ;

R 4-1 is halogen or hydroxyl;

R 4-2 is C 1 -C 4 alkyl;

R 4-3 is hydroxyl;

R 5 is H, hydroxyl or C 1 -C 4 alkoxy.
The pyrazoloquinazoline compound represented by formula I as claimed in claim 5, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof, wherein R 4 Selected from the group consisting of: cyano,
The pyrazoloquinazoline compound represented by formula I as claimed in claim 1, a pharmaceutically acceptable salt thereof, a solvate or a solvate of a pharmaceutically acceptable salt thereof, which has the following Mode:
The pyrazoloquinazoline compound represented by formula I as claimed in claim 7, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof, wherein R 1 is C 1 -C 4 alkoxy substituted by one or more R 1-1s.
The pyrazoloquinazoline compound represented by formula I as claimed in claim 7, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof, wherein R 2 is H or halogen.
The pyrazoloquinazoline compound represented by formula I as claimed in claim 7, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof, wherein R 4 Selected from the group consisting of: cyano,
A pyrazoloquinazoline compound, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof, characterized in that the pyrazoloquinazoline compound Choose from any of the following:
A preparation method of a pyrazoloquinazoline compound as shown in formula I, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof, comprising the following steps , in an organic solvent, under the action of a palladium catalyst, in an alkali reagent, the compound 1g, the ligand reagent and R 3 -H are subjected to a coupling reaction to obtain compound I;

Wherein, the definitions of R 1 , R 2 , R 3 , R 4 or R 5 are as described in claims 1 to 6; X is halogen.
The preparation method of claim 12, wherein the palladium-based catalyst is dipalladium tridibenzylidene acetone;

And/or, described organic solvent is tetradioxane;

And/or, described alkali reagent is cesium carbonate;

And/or, the ligand reagent is 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene.
A pyrazoloquinazoline compound represented by formula I, a pharmaceutically acceptable salt thereof, a solvate or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 11 The application of the solvate in the preparation of PLK1 inhibitor; the inhibitor is preferably an inhibitor used in vitro.
A pharmaceutical composition is characterized in that, it comprises the pyrazoloquinazoline compound shown in formula I as claimed in claims 1-11, its pharmaceutically acceptable salt, its solvate or its Solvates of pharmaceutically acceptable salts and pharmaceutical excipients.
A pyrazoloquinazoline compound as claimed in any one of claims 1 to 11 as shown in formula I, a pharmaceutically acceptable salt thereof, a solvate thereof or a pharmaceutically acceptable salt thereof The application of solvate in the preparation of medicine;

The medicament is a medicament for treating at least one of the following diseases: breast cancer, prostate cancer, lung cancer, colorectal cancer, liver cancer, pancreatic cancer, gastric cancer, esophageal cancer, melanoma, multiple myeloma, leukemia and lymphoma.
A pyrazoloquinazoline compound as shown in formula II, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof;

wherein, R 1 is C 1 -C 4 alkoxy substituted by one or more R 1-1 ; R 1-1 is independently halogen;

R 2 is H, halogen or C 1 -C 4 alkyl;

R 3' is a 7- to 9-membered heterocycloalkyl substituted or unsubstituted by one R 3-1 ; the 7- to 9-membered heterocycloalkyl is a 7- to 9-membered heterospirocycloalkyl or a 7- to 9-membered heterocycloalkyl Bridged cycloalkyl; heteroatoms are selected from one or more of N, O and S, and the number of heteroatoms is 1-2; R 3-1 is C 1 -C 4 alkyl;

R 4' is independently C 1 -C 4 alkyl substituted with one R 4-3 ; R 4-3 is hydroxy or halogen;

R 5 is H, hydroxy, C 1 -C 4 alkyl or C 1 -C 4 alkoxy.
The preparation method of the pyrazoloquinazoline compound as shown in claim 17, its pharmaceutically acceptable salt, its solvate or its pharmaceutically acceptable salt solvate, It is characterized in that R 1-1 is F, Cl, Br or I, such as F;

And/or, when R 1 is C 1 -C 4 alkoxy substituted by one or more R 1-1 , the C 1 -C 4 alkoxy is methoxy, ethoxy, iso- Propoxy or tert-butoxy, such as methoxy;

And/or, when R 2 is a C 1 -C 4 alkyl group, the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl such as methyl;

And/or, R 3-1 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl;

And/or, when R 4-3 is halogen, the halogen is F, Cl, Br or I;

And/or, when R 4' is independently a C 1 -C 4 alkyl group substituted by one R 4-3 , the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl propyl, n-butyl, isobutyl or tert-butyl, for example methyl;

And/or, when R 5 is a C 1 -C 4 alkyl group, the C 1 -C 4 alkyl group is methyl, ethyl, isopropyl or tert-butyl, such as methyl or ethyl;

And/or, when R 5 is C 1 -C 4 alkoxy, the C 1 -C 4 alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, such as methyl Oxygen.
The pyrazoloquinazoline compound represented by formula II, its pharmaceutically acceptable salt, its solvate or the solvate of its pharmaceutically acceptable salt as claimed in claim 17, characterized in that , R1 is
E.g

And/or, when R 3' is a 7- to 9-membered heterospirocycloalkyl group, the 7- to 9-membered heterospirocycloalkyl group is a 7- to 9-membered heterospirocycloalkyl group containing two nitrogen atoms, for example

And/or, when R 3' is a 7-9 membered heterospirocycloalkyl substituted by one R 3-1 , the 7-9 membered heterospirocycloalkyl substituted or unsubstituted by one R 3-1 is a 7- to 9-membered heterospirocycloalkyl group substituted with one R 3-1 containing two nitrogen atoms, such as

And/or, when R 3' is a 7- to 9-membered hetero-bridged cycloalkyl group, the 7- to 9-membered hetero-bridged cycloalkyl group is a 7- to 9-membered hetero-bridged cycloalkyl group containing two Ns, for example

And/or, when R 3' is a 7- to 9-membered hetero-bridged cycloalkyl substituted by one R 3-1 , the 7- to 9-membered hetero-bridged cycloalkyl substituted by one R 3-1 is a A 7- to 9-membered heterobridged cycloalkyl group of two nitrogens substituted by one R 3-1 , for example

and/or, R4 is -CH2OH .
The pyrazoloquinazoline compound represented by formula II, its pharmaceutically acceptable salt, its solvate or the solvate of its pharmaceutically acceptable salt as claimed in claim 17, characterized in that , the pyrazoloquinazoline compound shown in formula II, its pharmaceutically acceptable salt, its solvate or the solvate of its pharmaceutically acceptable salt can be shown as any one of the following:
A preparation method of a pyrazoloquinazoline compound as shown in formula II, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof, comprising the following steps , in an organic solvent, in an alkali reagent, under the action of a palladium catalyst, the compound 1g, the ligand reagent and R 3 -H are subjected to a coupling reaction to obtain compound II;

Wherein, the definitions of R 1 , R 2 , R 3' , R 4' or R 5 are as described in claims 12-15; X is halogen.
The preparation method of claim 21, wherein the palladium-based catalyst is tridibenzylideneacetone dipalladium;

And/or, described organic solvent is tetradioxane;

And/or, described alkali reagent is cesium carbonate;

And/or, the ligand reagent is 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene.
A compound of formula III, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof:

Wherein, R 1 is C 1 -C 4 alkoxy, C 1 -C 4 alkoxy substituted by one or more R 1-1 , and R 1- 1 is independently halogen or 3-6 membered cycloalkane;

R 2 is H, halogen or C 1 -C 4 alkyl;

R 3 is a 5- to 10-membered heterocycloalkyl group or a 5- to 10-membered heterocycloalkyl group substituted by one or more R 3-1 ; in the 5- to 10-membered heterocycloalkyl group, the heteroatom is selected from N One or more of , O and S, the number of heteroatoms is 1-2; R 3-1 is independently C 1 -C 4 alkyl;

R 4 is cyano, 3-6 membered cycloalkane, 3-6 membered cycloalkane substituted by one or more R 4-1 , 3-6 membered heterocycloalkane or substituted by one or more R 4-2 3-6-membered heterocycloalkane; C 1 -C 4 alkyl substituted by one or more R 4-3 ; in the 3-6 membered heterocycloalkyl, the heteroatom is selected from N, O and S One or more of , the number of heteroatoms is 1-2; R 4-1 is independently halogen, hydroxyl or C 1 -C 4 alkyl; R 4-2 is independently halogen or C 1 -C 4 alkyl; R 4-3 is independently halogen, cyano or hydroxyl;

R 5 is H, hydroxy C 1 -C 4 alkyl or C 1 -C 4 alkoxy.
The compound of formula III, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof according to claim 23, wherein R 1-1 is independently is F, Cl, Br or I, such as F;

And/or, R 1-1 is independently a 3- to 6-membered cycloalkane that is cyclopropane, cyclobutane or cyclopentane, such as cyclopropane or cyclobutane;

And/or, when R 1 is C 1 -C 4 alkoxy, the C 1 -C 4 alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, such as methyl Oxygen;

And/or, when R 1 is C 1 -C 4 alkoxy substituted by one or more R 1-1 , the C 1 -C 4 alkoxy is methoxy, ethoxy, iso- Propoxy or tert-butoxy, such as methoxy;

And/or, when R 2 is halogen, the halogen is F, Cl, Br or I;

And/or, when R 2 is a C 1 -C 4 alkyl group, the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl such as methyl;

And/or, when R 3-1 is a C 1 -C 4 alkyl group, the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl radical or tert-butyl, such as methyl;

And/or, when R 3 is a 5- to 10-membered heterocycloalkyl substituted by one or more R 3-1 , the 5- to 10-membered heterocycloalkyl is a 6- to 9-membered heterocycloalkyl, For example, a 6- to 9-membered heterocycloalkyl group containing two Ns, such as piperazinyl, hexahydropyridazinyl, hexahydropyrimidinyl, 3,8-diazabicyclo[3.2.1]octyl, octanyl Hydropyrrole[1,2-a]pyrazinyl, 2,6-diazaspiro[3.4]octyl, 3,6-diazabicyclo[3.2.0]heptyl, 1,6-bis azaspiro[3.4]octyl or octahydropyrrole[3,4-c]pyrrolyl;

And/or, when R 4-1 is halogen, the halogen is F, Cl, Br or I, such as F;

And/or, when R 4-1 is a C 1 -C 4 alkyl group, the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl radical or tert-butyl, such as methyl or ethyl;

And/or, when R 4-2 is a C 1 -C 4 alkyl group, the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl radical or tert-butyl, such as methyl or ethyl;

And/or, when R 4-2 is halogen, the halogen is F, Cl, Br or I, such as F;

And/or, when R 4-3 is halogen, the halogen is F, Cl, Br or I, such as F;

And/or, when R 4 is 3-6 membered cycloalkane, the 3-6 membered cycloalkane is cyclopropane, cyclobutane or cyclopentane, such as cyclopropane or cyclobutane;

And/or, when R 4 is a 3- to 6-membered cycloalkane substituted by one or more R 4-1 , the 3- to 6-membered cycloalkane is cyclopropane, cyclobutane or cyclopentane, such as cycloalkane Propane or cyclobutane;

And/or, when R 4 is a 3-6 membered heterocycloalkane substituted by one or more R 4-2 , the 3-6 membered heterocycloalkane is a 4-membered heterocycloalkane containing one N or O or a 5-membered heterocycloalkane containing one N or O;

And/or, when R 5 is a C 1 -C 4 alkyl group, the C 1 -C 4 alkyl group is methyl, ethyl, isopropyl or tert-butyl, such as methyl or ethyl;

And/or, when R 5 is C 1 -C 4 alkoxy, the C 1 -C 4 alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, such as methyl Oxygen.
The compound of formula III, its pharmaceutically acceptable salt, its solvate or its pharmaceutically acceptable salt solvate according to claim 23, wherein said R 1 is
E.g

and/or, R3 is

and/or, R3 is

and/or, R3 is

And/or, when R 4 is a 3-6 membered cycloalkane substituted by one or more R 4-1 , the 3-6 membered cycloalkane substituted by one or more R 4-1 is:

And/or, when R 4 is a 3- to 6-membered heterocycloalkane, the 3- to 6-membered heterocycloalkane is

And/or, when R 4 is a 3-6 membered heterocycloalkane substituted by one or more R 4-2 , the 3-6 membered heterocycloalkane substituted by one or more R 4-2 is

And/or, when R 4 is C 1 -C 4 alkyl substituted by one or more R 4-3 , the C 1 -C 4 alkyl substituted by one or more R 4-3 is
The compound of formula III, its pharmaceutically acceptable salt, its solvate or its pharmaceutically acceptable salt solvate of claim 23, wherein R 4 is cyano, 3-6-membered cycloalkane, 3-6 membered heterocycloalkane substituted by one or more R 4-1 , or 3-6 membered heterocycloalkane substituted by one or more R 4-2 , or substituted by one or more C 1 -C 4 alkyl substituted with one R 4-3 ;

And/or, R 4-1 is halogen or hydroxyl;

And/or, R 4-2 is C 1 -C 4 alkyl;

And/or, R 4-3 is hydroxyl;

And/or, R 5 is H, hydroxy or C 1 -C 4 alkoxy.
The compound of formula III, its pharmaceutically acceptable salt, its solvate or its pharmaceutically acceptable salt solvate of claim 23, wherein R 4 is cyano, 3-6 membered cycloalkane substituted by one or more R 4-1 , 3-6 membered heterocycloalkane, 3-6 membered heterocycloalkane substituted by one or more R 4-2 , or substituted by one or more C 1 -C 4 alkyl substituted with one R 4-3 ;

R 4-1 is halogen or hydroxyl;

R 4-2 is C 1 -C 4 alkyl;

R 4-3 is hydroxyl;

R 5 is H, hydroxyl or C 1 -C 4 alkoxy.
The compound of formula III, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof according to claim 23, wherein the compound is:
Use of the compound according to any one of claims 1 to 28, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof in the preparation of a PLK1 inhibitor; the The inhibitor is preferably an inhibitor used in vitro.
A pharmaceutical composition comprising the compound according to any one of claims 1 to 28, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof, and a pharmaceutical excipient .
Use of the compound according to any one of claims 1 to 28, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof in the preparation of a medicament;

The medicine is a medicine for treating at least one of the following diseases: breast cancer, prostate cancer, lung cancer, colorectal cancer, liver cancer, pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, melanoma, osteosarcoma, multiple myeloma , leukemia and lymphoma.
A pharmaceutical combination comprising a compound according to any one of claims 1 to 28, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof, and one or Multiple chemotherapeutic agents, the one or more chemotherapeutic agents being used simultaneously, separately or sequentially with the compound, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof.