CN116528871A

CN116528871A - Pyrazoloquinazoline compound, and preparation method and application thereof

Info

Publication number: CN116528871A
Application number: CN202180078594.6A
Authority: CN
Inventors: 魏钊; 王文义; 崔洪; 李元念; 周红; 石峰
Original assignee: Hengyuan Biomedical Technology Suzhou Co ltd
Current assignee: Hengyuan Biomedical Technology Suzhou Co ltd
Priority date: 2020-12-31
Filing date: 2021-12-28
Publication date: 2023-08-01
Also published as: WO2022143576A1; TW202241413A

Abstract

Relates to pyrazoloquinazoline compounds, a preparation method and application thereof. In particular to pyrazoloquinazoline compounds shown as a formula I, pharmaceutically acceptable salts thereof, solvates thereof or solvates of pharmaceutically acceptable salts thereof. The compound has obvious PLK1 inhibiting effect.

Description

Pyrazoloquinazoline compound, and preparation method and application thereof

Technical Field

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

Background

PLKs (Polo-like kinases) belong to serine/threonine kinases. Up to now, a total of 5 subtypes, PLK1-5, are found in the family. PLKs are expressed primarily in dividing cells and play a vital role in control of the cell cycle and mitosis. Among the PLK families, PLK1 is the most studied and most elaborated, and its biological activity is also most clearly described. PLK1 is widely recognized by the scientific community as a key kinase directing mitotic entry, centrosome maturation and separation, bipolar spindle formation, metaphase to late transition, and cytokinesis initiation. It was found that PLK1 is highly expressed not only in various tumor tissues but also activated in DNA damage and replication stress, which is also consistent with abnormal activity of proliferation of tumor cells. The PLK1 gene can be knocked out through chemical small molecule inhibitors or biological means, so that a great deal of tumor cells can be found to stay in the G2/M phase, and further the apoptosis of the tumor cells is caused. Thus, PLK1 is considered a promising antitumor target, and in particular its highly expressed properties in various tumor tissues make it possible to have a broad spectrum of antitumor potential. In addition, the expression level of PLK1 in normal cells is low, so that the development of PLK1 inhibitors as antitumor drugs may have a good safety window.

Compared with high expression of PLK1 in tumor cells, PLK2 and PLK3 are more expressed in cells such as nerve cells after mitosis, so that the development of PLK1 specific inhibitors may avoid toxic and side effects caused by simultaneous inhibition of PLK 2/3. The PLK1 inhibitor Volasertib (BI-6727) developed by BI (Boehringer Ingelheim) has serious toxic and side effects in clinic due to lack of selectivity to PLK2/3, and limits the curative effect, so that the third phase clinical failure is caused. As PLK1 specific inhibitor, the small molecule inhibitor, novansertib, developed by cardioff Oncology company, showed better antitumor activity and controllable toxic side effects in a number of second-phase clinical trials. However, onvansertib still has more room for improvement, such as poor transmembrane properties, resulting in lower bioavailability. Moreover, due to its longer half-life (t _1/2 Greater than 24 hours) results in only intermittent dosing during the treatment cycle, further limiting the therapeutic effect that can be achieved, in addition to rendering it more pronounced toxic side effects.

Disclosure of Invention

The invention aims to solve the technical problems of single structure, poor inhibition effect and the like of PLK1 inhibitors in the prior art, and provides a pyrazoloquinazoline compound, a preparation method and application thereof, wherein the compound has an obvious inhibition effect on PLK 1.

The invention solves the technical problems through the following technical proposal.

The invention provides pyrazoloquinazoline compounds shown as a formula I, pharmaceutically acceptable salts thereof, solvates thereof or solvates of pharmaceutically acceptable salts thereof;

wherein R is ¹ Is C ₁ ～C ₄ Alkoxy, substituted by one or more R ^1-1 Substituted C ₁ ～C ₄ Alkoxy or 3-6 membered cycloalkyloxy; r is R ^1-1 Independently halogen or 3-6 membered cycloalkane;

R ² is H, halogen or C ₁ ～C ₄ An alkyl group;

R ³ is 5-10 membered heterocycloalkyl or substituted by one or more R ^3-1 Substituted 5-to 10-membered heterocycloalkyl; in the 5-10 membered heterocycloalkyl, the hetero atom is one or more selected from N, O and S, and the number of the hetero atom is 1-2; r is R ^3-1 Independently C ₁ ～C ₄ An alkyl group;

R ⁴ is cyano, 3-6 membered cycloalkane, substituted with one or more R ^4-1 Substituted 3-to 6-membered cycloalkanes, 3-to 6-membered heterocycloalkanes or by one or more R ^4-2 Substituted 3-6 membered heterocycloalkanes; is/are R ^4-3 Substituted C ₁ ～C ₄ An alkyl group; in the 3-6 membered heterocycloalkyl, the hetero atom is one or more selected from N, O and S, and the number of the hetero atom is 1-2; r is R ^4-1 Independently halogen, hydroxy or C ₁ ～C ₄ An alkyl group; r is R ^4-2 Independently halogen or C ₁ ～C ₄ An alkyl group; r is R ^4-3 Independently halogen, cyano or hydroxy;

R ⁵ Is H, hydroxy, C ₁ ～C ₄ Alkyl or C ₁ ～C ₄ An alkoxy group.

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

in one embodiment, R ^1-1 Independently F, cl, br or I, e.g., F.

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

In one embodiment, when R ¹ Is C ₁ ～C ₄ In the case of alkoxy, said C ₁ ～C ₄ Alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, for example methoxy.

In one embodiment, when R ¹ To be covered by one or more R ^1-1 Substituted C ₁ ～C ₄ In the case of alkoxy, said C ₁ ～C ₄ Alkoxy can be methoxy, ethoxy, isopropoxy or tert-butoxy, for example methoxy.

In one embodiment, when R ¹ In the case of 3-to 6-membered cycloalkyloxy, the 3-to 6-membered cycloalkoxy group is cyclopropyloxy, cyclobutyloxy or cyclopentyloxy, for example cyclopropyloxy or cyclobutyloxy.

In one embodiment, R ¹ (may be) For example

In one embodiment, when R ² In the case of halogen, the halogen may be F, cl, br or I.

In one embodiment, when R ² Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl may be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl.

In one embodiment, when R ^3-1 Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl may be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl.

In one embodiment, when R ³ To be covered by one or more R ^3-1 In the case of substituted 5-to 10-membered heterocycloalkyl, the 5-to 10-membered heterocycloalkyl may be 6-to 9-membered heterocycloalkyl, for example 6-to 9-membered heterocycloalkyl containing two N, for 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 octahydro-pyrrole [3,4-c ]]Pyrrole groups.

In one embodiment, R ³ Preferably is (b is ANDIs a site of attachment).

In one embodiment, when R ³ (may be)

In one embodiment, R ³ (may be)

In one embodiment, R ⁴ Is cyano, covered by one or more R ^4-1 Substituted 3-to 6-membered cycloalkanes, 3-to 6-membered heterocycloalkanes or by one or more R ^4-2 Substituted 3-6 membered heterocycloalkanes.

In one embodiment, R ^4-1 Is halogen or hydroxy.

In one embodiment, R ^4-2 Is C ₁ ～C ₄ An alkyl group.

In one embodiment, R ⁵ H.

In one embodiment, when R ^4-1 In the case of halogen, the halogen is F, cl, br or I, such as F.

In one embodiment, when R ^4-1 Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ The alkyl group may be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl or ethyl.

In one embodiment, when R ^4-2 Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ The alkyl group may be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl or ethyl.

In one embodiment, when R ^4-2 In the case of halogen, the halogen is F, cl, br or I, such as F.

In one embodiment, when R ⁴ In the case of 3-to 6-membered cycloalkanes, the 3-to 6-membered cycloalkanes may be cyclopropane, cyclobutane or cyclopentane, for example cyclopropane or cyclobutane.

In one embodiment, when R ⁴ To be covered by one or more R ^4-1 In the case of substituted 3-to 6-membered cycloalkanes, the 3-to 6-membered cycloalkane may be cyclopropane, cyclobutane or cyclopentane, for example cyclopropane or cyclobutane.

In one embodiment, when R ⁴ To be covered by one or more R ^4-1 In the case of substituted 3-to 6-membered cycloalkanes, said cycloalkane is substituted with one R ^4-1 The substituted 3-to 6-membered cycloalkane may be

In one embodiment, when R ⁴ In the case of 3-to 6-membered heterocyclic alkane, the 3-to 6-membered heterocyclic alkane may be

In one embodiment, when R ⁴ Is covered by one R ^4-2 In the case of substituted 3-6 membered heterocycloalkyl, the 3-6 membered heterocycloalkyl may be a 4 membered heterocycloalkyl containing one N or O or a 5 membered heterocycloalkyl containing one N or O.

In one embodiment, when R ⁴ To be covered by one or more R ^4-2 In the case of substituted 3-to 6-membered heterocycloalkanes, said substituted 3-to 6-membered heterocycloalkanes are substituted by one or more R ^4-2 Substituted 3-to 6-membered heterocycloalkanes

In one embodiment, when R ⁴ To be covered by one or more R ^4-3 Substituted C ₁ ～C ₄ In the case of alkyl groups, said groups are substituted by one or more R ^4-3 Substituted C ₁ ～C ₄ Alkyl is

In one embodiment, R ⁴ Is cyano, covered by one or more R ^4-1 Substituted 3-to 6-membered cycloalkanes, 3-to 6-membered heterocycloalkanes or by one or more R ^4-2 Substituted 3-6 membered heterocycloalkyl, or by one or more R ^4- ³ Substituted C ₁ ～C ₄ An alkyl group.

In one embodiment, R ⁴ To be covered by one or more R ^4-1 Substituted 3-6 membered cycloalkanes, wherein R ^4-1 Is halogen or hydroxy.

In one embodiment, R ⁴ To be covered by one or more R ^4-2 Substituted 3-6 membered heterocycloalkanes, wherein R ^4- ² Is C ₁ ～C ₄ An alkyl group.

In one embodiment, R ⁴ To be covered by one or more R ^4-3 Substituted C ₁ ～C ₄ Alkyl, wherein R is ^4-3 Is hydroxyl.

In one embodiment, R ⁴ Selected from the group consisting of: cyano group,

In one embodiment, R ⁵ Is H or hydroxy.

In one embodiment, R ⁵ Is C ₁ ～C ₄ Alkyl, said C ₁ ～C ₄ The alkyl group may be methyl, ethyl, isopropyl or tert-butyl, for example methyl or ethyl.

In one embodiment, R ⁵ Is C ₁ ～C ₄ Alkoxy, said C ₁ ～C ₄ Alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, e.g. methoxy

The invention also provides pyrazoloquinazoline compounds shown as formula Ia, pharmaceutically acceptable salts thereof, solvates thereof or solvates of pharmaceutically acceptable salts thereof:

in one embodiment, the invention provides pyrazoloquinazolines, pharmaceutically acceptable salts thereof, solvates thereof, or solvates of pharmaceutically acceptable salts thereof, as shown in any one of the following:

the invention also provides pyrazoloquinazoline compounds shown as the formula II, pharmaceutically acceptable salts thereof, solvates thereof or solvates of pharmaceutically acceptable salts thereof:

Wherein R is ¹ To be covered by one or more R ^1-1 Substituted C ₁ ～C ₄ An alkoxy group; r is R ^1-1 Independently halogen;

R ² is H, halogen or C ₁ ～C ₄ An alkyl group;

R ^3’ is 7-9 membered heterocycloalkyl or is substituted by one R ^3-1 Substituted 7-to 9-membered heterocycloalkyl; the 7-9 membered heterocycloalkyl is 7-9 membered hetero spirocycloalkyl or 7-9 membered hetero bridged cycloalkyl; said quilt is R ^3-1 Substituted 7-to 9-membered heterocycloalkyl being substituted by one R ^3-1 Substituted 7-to 9-membered heterospirocycloalkyl or by one R ^3-1 Substituted 7-9 membered heterobridged cycloalkyl; the hetero atoms are selected from one or more of N, O and S, and the number of the hetero atoms is 1-2; r is R ^3-1 Is C ₁ ～C ₄ An alkyl group;

R ^4’ independently is covered by one R ^4-3 Substituted C ₁ ～C ₄ An alkyl group; r is R ^4-3 Is hydroxy or halogen;

R ⁵ is H, hydroxy C ₁ ～C ₄ Alkyl or C ₁ ～C ₄ An alkoxy group.

In one embodiment, R ^1-1 F, cl, br or I, e.g., F.

In one embodiment, when R ¹ To be covered by one or more R ^1-1 Substituted C ₁ ～C ₄ Alkoxy groups, when said

In one embodiment, R ^3-1 May be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl.

In one embodiment, when R ^3’ In the case of a 7-to 9-membered heterospirocycloalkyl group, the 7-to 9-membered heterospirocycloalkyl group may be a 7-to 9-membered heterospirocycloalkyl group having two nitrogen atoms, for example

In one embodiment, when R ^3’ Is covered by one R ^3-1 In the case of substituted 7-to 9-membered heterospirocycloalkyl, said residue is substituted with one R ^3-1 The substituted or unsubstituted 7-to 9-membered heterospirocycloalkyl group may be substituted with one R having two nitrogen atoms ^3-1 Substituted 7-to 9-membered heterospirocycloalkyl groups, e.g.

In one embodiment, when R ^3’ In the case of a 7-to 9-membered heterobridged cycloalkyl group, the 7-to 9-membered heterobridged cycloalkyl group may be a 7-to 9-membered heterobridged cycloalkyl group containing two N's, for example

In one embodiment, when R ^3’ Is covered by one R ^3-1 In the case of substituted 7-to 9-membered heterobridged cycloalkyl, said member is substituted by oneR is a number of ^3-1 The substituted 7-to 9-membered heterobridged cycloalkyl group may be substituted with one R ^3-1 Substituted 7-to 9-membered heterobridged cycloalkyl radicals, e.g.

In one embodiment, when R ^4-3 In the case of halogen, the halogen is F, cl, br or I.

In one embodiment, when R ^4’ Independently is covered by one R ^4-3 Substituted C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl may be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl.

In one embodiment, R ⁴ Can be-CH ₂ OH。

In one embodiment, when R ⁵ Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ The alkyl group may be methyl, ethyl, isopropyl or tert-butyl, for example methyl or ethyl.

In one embodiment, when R ⁵ Is C ₁ ～C ₄ In the case of alkoxy, said C ₁ ～C ₄ Alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, for example methoxy.

In one embodiment, the pyrazoloquinazoline compound shown in formula II, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof may be any one of the following:

the invention also provides pyrazoloquinazoline compounds shown in a formula III, pharmaceutically acceptable salts thereof, solvates thereof or solvates of the pharmaceutically acceptable salts thereof:

wherein R is ¹ Is C ₁ ～C ₄ Alkoxy or by one or more R ^1-1 Substituted C ₁ ～C ₄ An alkoxy group; r is R ^1-1 Independently halogen or 3-6 membered cycloalkane;

R ² is H, halogen or C ₁ ～C ₄ An alkyl group;

R ⁵ is H, hydroxy, C ₁ ～C ₄ Alkyl or C ₁ ～C ₄ An alkoxy group.

In one embodiment, R ¹ Is C ₁ ～C ₄ Alkoxy, such as methoxy, ethoxy, isopropoxy or tert-butoxy.

In one embodiment, R ¹ To be covered by one or more R ^1-1 Substituted C ₁ ～C ₄ Alkoxy, wherein said C ₁ ～C ₄ Alkoxy can be methoxy, ethoxy, isopropoxy or tert-butoxy, for example methoxy.

In one embodiment, R ¹ To be covered by one or more R ^1-1 Substituted C ₁ ～C ₄ Alkoxy, wherein R is ^1-1 Independently F, cl, br or I, e.g., F.

In one embodiment, R ¹ Can be selected from

In one embodiment, R ² H.

In one embodiment, R ² Is halogen, wherein the halogen is F, cl, br or I.

In one embodiment, R ² Is C ₁ ～C ₄ Alkyl groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

In one embodiment, R ³ 5-to 10-membered heterocycloalkyl, e.g. 6-to 9-membered heterocycloalkyl containing two N, e.g. 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 octahydro-pyrrole [3,4-c ]]Pyrrole groups.

In one embodiment, R ³ To be covered by one or more R ^3-1 Substituted 5-to 10-membered heterocycloalkyl, wherein the 5-to 10-membered heterocycloalkyl is, for example, 6-to 9-membered heterocycloalkyl containing two N, for 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 octahydro-pyrrole [3,4-c ]]Pyrrolyl, wherein R is ^3-1 Is C ₁ ～C ₄ Alkyl groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

In one embodiment, R ³ Is that

In one embodiment, R ⁴ Is cyano.

In one embodiment, R ⁴ Is a 3-to 6-membered cycloalkane, such as cyclopropane, cyclobutane or cyclopentane.

In one embodiment, R ⁴ To be covered by one or more R ^4-1 Substituted 3-6 membered cycloalkane, wherein the 3-6 membered cycloalkane is for example cyclopropane, cyclobutane or cyclopentane, R ^4-1 Halogen, such as F, cl, br or I.

In one embodiment, R ⁴ To be covered by one or more R ^4-1 Substituted 3-6 membered cycloalkane, wherein the 3-6 membered cycloalkane is for example cyclopropane, cyclobutane or cyclopentane, R ^4-1 Is C ₁ ～C ₄ Alkyl groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

In one embodiment, R ⁴ Is that

In one embodiment, R ⁴ Is a 3-6 membered heterocycloalkyl, for example a 4 membered heterocycloalkyl containing one N or O or a 5 membered heterocycloalkyl containing one N or O.

In one embodiment, R ⁴ To be covered by one or more R ^4-2 Substituted 3-6 membered heterocycloalkanes, e.g. 4-membered heterocycloalkanes comprising one N or O or 5-membered heterocycloalkanes comprising one N or O, where R ^4-2 Halogen, such as F, cl, br or I.

In one embodiment, R ⁴ Is that

In one embodiment, R ⁴ To be covered by one or more R ^4-2 Substituted 3-6 membered heterocycloalkanes, e.g. 4-membered heterocycloalkanes comprising one N or O or 5-membered heterocycloalkanes comprising one N or O, where R ^4-2 Is C ₁ ～C ₄ Alkyl groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

In one embodiment, R ⁴ To be covered by one or more R ^4-3 Substituted C ₁ ～C ₄ Alkyl, wherein the C ₁ ～C ₄ Alkyl is methyl, ethyl, isopropyl or tert-butyl, R ^4-3 Halogen, such as F, cl, br or I.

In one embodiment, R ⁴ To be covered by one or more R ^4-3 Substituted C ₁ ～C ₄ Alkyl, wherein the C ₁ ～C ₄ Alkyl is methyl, ethyl, isopropyl or tert-butyl, R ^4-3 Is cyano.

In one embodiment, R ⁴ To be covered by one or more R ^4-3 Substituted C ₁ ～C ₄ Alkyl, wherein the C ₁ ～C ₄ Alkyl is methyl, ethyl, isopropyl or tert-butyl, R ^4-3 Is hydroxyl.

In one embodiment, R ⁴ Is that

In one embodiment, R ⁵ Is H or hydroxy.

In one embodiment, R ⁵ Is C ₁ ～C ₄ Alkyl groups 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 pyrazoloquinazoline compound shown in the formula III is:

the invention also provides a preparation method of pyrazoloquinazoline compound shown as the formula I, pharmaceutically acceptable salt thereof, solvate thereof or solvate of pharmaceutically acceptable salt thereof, which comprises the following steps of adding 1g of compound, ligand reagent and R under the action of palladium catalyst in an organic solvent and alkali reagent ³ Carrying out coupling reaction on H to obtain a compound I;

wherein R is ¹ 、R ² 、R ³ 、R ⁴ Or R is ⁵ Is defined as above; x is halogen; l is hydrochloride.

In the coupling reaction, the palladium catalyst may be dibenzylidene acetone dipalladium.

In the coupling reaction, the organic solvent may be a conventional solvent for such a reaction in the art, for example, a tetradioxane.

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

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

The invention also provides pyrazoloquinazoline compounds shown as the formula II, pharmaceutically acceptable salts thereof, solvates thereof or solvates of pharmaceutically acceptable salts thereofComprises the steps of reacting 1g of a compound, a ligand reagent and R in an organic solvent under the action of a palladium catalyst ³ Carrying out coupling reaction on L to obtain a compound II;

wherein R is ¹ 、R ² 、R ^3’ 、R ^4’ Or R is ⁵ Is defined as above; x is halogen; l is hydrochloride.

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

The invention provides application of pyrazoloquinazoline compounds shown as the formula I, pharmaceutically acceptable salts thereof, solvates thereof or solvates of pharmaceutically acceptable salts thereof in preparation of PLK1 inhibitors; the inhibitor is preferably an inhibitor for use in vitro.

The invention also provides a pharmaceutical composition, which also comprises pyrazoloquinazoline compounds shown as a formula I, pharmaceutically acceptable salts thereof, solvates thereof or solvates of the pharmaceutically acceptable salts thereof and pharmaceutical excipients.

The invention also provides application of the pyrazoloquinazoline compound shown as the formula I, pharmaceutically acceptable salt thereof, solvate thereof or solvate of pharmaceutically acceptable salt thereof in preparation of medicines;

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

The invention provides application of pyrazoloquinazoline compounds shown as formula II, pharmaceutically acceptable salts thereof, solvates thereof or solvates of pharmaceutically acceptable salts thereof in preparation of PLK1 inhibitors; the inhibitor is preferably an inhibitor for use in vitro.

The invention also provides a pharmaceutical composition, which comprises pyrazoloquinazoline compounds shown as the formula I, pharmaceutically acceptable salts thereof, solvates thereof or solvates of the pharmaceutically acceptable salts thereof and pharmaceutical excipients.

The invention also provides application of the pyrazoloquinazoline compound shown as the formula II, pharmaceutically acceptable salt thereof, solvate thereof or solvate of pharmaceutically acceptable salt thereof in preparation of medicines;

the medicine is used 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 invention also provides a pharmaceutical composition comprising a compound of the invention, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof, and one or more chemotherapeutic agents.

In one embodiment, the one or more chemotherapeutic agents are used simultaneously, separately or sequentially with the compounds of the invention, pharmaceutically acceptable salts thereof, solvates thereof, or solvates of pharmaceutically acceptable salts thereof.

Interpretation of the terms

In the various parts of the present specification, substituents of the presently disclosed compounds are disclosed in terms of the type or scope of groups. It is specifically noted that the present invention includes each individual subcombination of the individual members of these group classes and ranges. For example, the term "C ₁ ～C ₄ Alkyl "refers specifically to independently disclosed methyl, ethyl, C ₃ Alkyl (i.e. propyl, including n-propyl and isopropyl), C ₄ Alkyl (i.e., butyl, including n-butyl, isobutyl, sec-butyl, and tert-butyl).

When none of the listed substituents indicates through which atom it is attached to a compound included in the chemical structural formula but not specifically mentioned, such substituents may be bonded through any of their atoms. Combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

Where no substituent is explicitly indicated in a recited group, such a group is merely unsubstituted. For example when "C ₁ ～C ₄ Alkyl "not previously" substituted or unsubstituted "refers only to" C ₁ ～C ₄ Alkyl "as such or" unsubstituted C ₁ ～C ₄ An alkyl group.

The term "C _i -C _j "means a range of carbon number where i and j are integers and j is greater than i, and the range of carbon number includes the endpoints (i.e., i and j) and each integer point between the endpoints. For example, C ₁ ～C ₆ Represents a 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, in particular 1 to 10, in particular 1 to 8, in particular 1 to 6, in particular 1 to 5, in particular 1 to 4, in particular 1 to 3, or in particular 1 to 2 carbon atoms.

The term "alkyl" refers to a saturated straight or branched hydrocarbon group. The term "C _i -C _j Alkyl "refers to an alkyl group having i to j carbon atoms. In some embodiments, the alkyl group comprises 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 5From 1 to 4 carbon atoms, from 1 to 3 carbon atoms, or from 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 (t-butyl), 1-pentyl (n-pentyl), 2-pentyl, 3-pentyl, 2-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, 3-dimethyl-2-butyl, 3-dimethyl-2-butyl, 1-heptyl, 1-octyl, and the like.

The term "alkoxy" refers to an alkyl group, as defined above, attached to the parent molecule through an oxygen atom. The term "C _i -C _j Alkoxy "means that the alkyl portion of the alkoxy group has i to j carbon atoms. In some embodiments, the alkoxy groups contain 1 to 12 carbon atoms. In some embodiments, the alkoxy groups contain 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, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms.

The term "cycloalkane" or "cycloalkyl" refers to a compound having the indicated number of carbon atoms (e.g., C ₃ ～C ₆ ) Saturated monocyclic ring groups consisting of carbon atoms only. Cycloalkyl includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

The term "heterocycloalkyl" refers to a cyclic group of a specified number of ring atoms (e.g., 5-10 membered), of a specified number of heteroatoms (e.g., 1, 2, or 3), of a specified heteroatom species (one or more of N, O and S), which is a single ring, bridged ring, or spiro ring, and each ring is saturated. Bridged ring means a polycyclic ring having two or more atoms in common between the monocyclic rings. Spiro refers to polycyclic rings that share one atom between the 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 that is reacted with a pharmaceutically acceptable (relatively non-toxic, safe, suitable for patient use) acid or base. When the compound contains a relatively acidic functional group, the base addition salt may 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 salts, and the like. When the compound contains a relatively basic functional group, the acid addition salt may 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 in particular Handbook of Pharmaceutical Salts Properties, selection, and Use (P.Heinrich Stahl, 2002).

The term "solvate" refers to a material formed upon crystallization of a compound with a solvent (including, but not limited to, water, methanol, ethanol, etc.). The solvates are divided into stoichiometric solvates and non-stoichiometric solvates.

The term "solvate of a pharmaceutically acceptable salt" refers to a compound formed by combining 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.), wherein the pharmaceutically acceptable salt is as defined above for the term "pharmaceutically acceptable salt" and the solvent is stoichiometric or non-stoichiometric. Solvates of the pharmaceutically acceptable salts include, but are not limited to, hydrochloride monohydrate.

The term "pharmaceutical excipients" refers to excipients and additives used in the manufacture of medicaments and formulation of prescriptions, and is all matter contained in the pharmaceutical formulation except for the active ingredient. See in particular the pharmacopoeia of the people's republic of China (2020 edition) or Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009).

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.

As used herein, the singular forms "a", "an", and "the" are understood to include plural referents unless the context clearly dictates otherwise. Furthermore, the term "comprising" is an open-ended limitation and does not exclude other aspects, i.e. it includes the content indicated by the invention.

Unless otherwise indicated, the present invention employs conventional methods of mass spectrometry, elemental analysis, and the various steps and conditions are referred to in the art by conventional procedures and conditions.

The present invention employs, unless otherwise indicated, standard nomenclature for analytical chemistry, organic synthetic chemistry and optics, and standard laboratory procedures and techniques. In some cases, standard techniques are used for chemical synthesis, chemical analysis, and light emitting device performance detection.

In addition, unless explicitly indicated otherwise, the description of the invention as "…" independently is to be understood broadly as meaning that each individual described may be independent of the other, and may be the same or different. In more detail, the description "… is independently" may mean that specific options expressed between the same symbols in different groups do not affect each other; it may also be expressed that specific options expressed between the same symbols in the same group do not affect each other.

Those skilled in the art will appreciate that, in accordance with convention used in the art, the present application describes the structural formula of a group as used inMeaning that the corresponding group is linked to other fragments, groups in the compound through this site.

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 invention provides a pyrazoloquinazoline compound, a preparation method and application thereof, and the pyrazoloquinazoline compound has at least any one of the following advantages that the PLK1 inhibition effect is obvious, and the half life of the medicament can be further regulated.

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

First step

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

Second step

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℃under nitrogen and reacted for 2 hours. The reaction was completed. After cooling, water (50 mL) was added to the reaction mixture, ethyl acetate was extracted (20 mL. Times.2), and the organic phase was washed successively with water (20 mL. Times.1) and saturated brine (20 mL. Times.1), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 1c (750 mg, yield: 76%).

Third step

A solution of compound 1c (750 mg,2.4 mmol) and compound 1d (428 mg,2.4 mmol) in dimethylformamide (10 mL) was heated to 120deg.C under nitrogen and stirred for 2 hours. The reaction was completed. Cooled, water 30 (mL) was added, ethyl acetate (20 mL x 2) was extracted, and the organic phase was washed successively with water (20 mL x 1) and saturated brine (20 mL x 1), dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product which was subjected to silica gel column chromatography (methanol/dichloromethane=0/100%) to give compound 1e (780 mg, yield: 59%).

LCMS(M+H)+m/z：542/544。

Fourth step

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

Fifth step

To a solution of compound 1f (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-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (684 mg,1.8 mmol) under nitrogen at room temperature, and the mixture was reacted for 2 hours. The reaction was completed. The reaction solution was diluted with water (20 mL), extracted with ethyl acetate (20 mL. Times.3), and the organic phase was washed successively with water (10 mL. Times.1) and saturated brine (10 mL. Times.1), dried over anhydrous sodium sulfate, filtered, and concentrated to give 1g of the compound (510 mg, yield: 83%).

LCMS(M+H)+m/z：513/515。

Sixth step

A solution of 1g (51 mg,0.1 mmol) of 8-methyl-3, 8-diazabicyclo [3.2.1] octane dihydrochloride (24 mg,0.12 mmol), cesium carbonate (130 mg,0.4 mmol), 4, 5-bis (diphenylphosphine) -9, 9-dimethylxanthene (24 mg,0.04 mmol) and tris (dibenzylideneacetone) dipalladium (18 mg,0.02 mmol) in 1, 4-dioxane (2 mL) was heated to reflux under nitrogen for 4 hours. The reaction was completed. Cooled, water (10 mL) was added to the reaction mixture, ethyl acetate (15 mL. Times.3) was extracted, and the organic phase was successively washed with water (10 mL. Times.1) and saturated brine (10 mL. Times.1), dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product, which was purified by preparative HPLC to give compound 1 (1.5 mg, yield: 27%).

LCMS(M+H)+m/z：559。

Example 2

First step

Compound 2 (3.0 mg) was obtained from 1g of compound according to the method of example 1.

LCMS(M+H)+m/z：559。

Example 3

First step

Compound 3 (1.6 mg) was obtained from 1g of compound according to the method of example 1.

LCMS(M+H)+m/z：545。

Example 4

First step

Compound 4 (1.3 mg) was obtained from 1g of compound according to the method of example 1.

LCMS(M+H)+m/z：545。

Example 5

First step

Compound 5 (2.0 mg) was obtained from compound 1g according to the method of example 1.

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). Para-toluene sulfonic acid (15.4 g,89.2 mmol) was added and the solution stirred at 100℃for 48 hours. The solvent was evaporated and the residue was dissolved with DCM (500 mL) and NaHCO ₃ Is washed with a saturated solution (500 mL). Na for organic phase ₂ SO ₄ Drying and concentrating. Purification by silica gel column chromatography (0% to 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

At N ₂ LiHMDS (51 mL,51.0mmol,1M in THF) was added dropwise to a solution of 2-methoxycyclohexyl-2-en-1-one (6.40 g,45.7 mmol) in THF (60 mL) at-50deg.C under an atmosphere. After stirring for 30 minutes at-50℃diethyl oxalate (7.47 g,51.0 mmol) was added. The solution was stirred at 25℃for 16 hours. After the reaction was completed, water (100 mL) was added, pH was adjusted to 4-5 by adding 1N HCl, and the resulting solution was extracted with EA (50 mL. Times.3). Na for organic layer ₂ SO ₄ Dried and evaporated to dryness. The crude product was purified by column (0% to 15% ea in PE) to give 2- (3-methoxy-2-oxocyclohex-3-en-1-yl) -2-oxoacetic acid ethyl ester 6f_3 (8.2 g, 71.4%) as a pale yellow oil. 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 a solution of 2- (3-ethoxy-2-oxocyclohex-3-en-1-yl) -2-oxoacetic acid ethyl ester 6F_3 (3.0 g,0.012 mmol) in ethanol (30 mL) was added hydroxyethylhydrazine (0.6 mL,0.012 mmol) at 25deg.C. The reaction was stirred at 80℃for 5 hours. The solvent was then evaporated and the residue redissolved with DCM (30 mL). The organic layer was washed with water, with Na ₂ SO ₄ Drying and concentrating. The residue was dissolved in THF (12 mL) and HCl (12 mL,1 mol/L) and stirred at 25℃for 2 hours, the reaction was run with saturated NaHCO ₃ The solution (50 mL) was quenched with EA (50 ml×3) and the combined organic layers were 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-1H-indazole-3-carboxylic acid ethyl ester 6f_3 (2.4 g, crude) as a brown oil. ESI-MS (M+H) ⁺ ＝253。

Fourth step

6F_3 (4 g,15.9 mmol) was dissolved in DMF (24 mL) at 25℃and then 8mL of DMFDMA was added. The resulting mixture was stirred at 80 ℃ for 20h, then cooled to 25 ℃, concentrated to remove the 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 (600 MHz, 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).

Fifth step

5-bromo-4-fluoro-2-methoxyaniline 6A (7.5 g,34.3 mmol) was dissolved in EtOH (90 mL) and Ac was added ₂ O (8.7 g,85.6 mmol). The mixture was stirred at 25℃for 2.5h 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。

Sixth step

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) were added to the round bottom flask. 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℃for 1h. The reaction mixture was added saturated aqueous NH4Cl (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% to 5% methanol, dichloromethane) to give N- (4-fluoro-2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) acetamide 6a_2 (400 mg, 24.8%) as a pale yellow solid. LCMS (m+h) +=282.

Seventh step

N- (4-fluoro)-2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) acetamido 6A_2 (400 mg,1.67 mmol) was dissolved in EtOH (4 mL), stirred at room temperature, and HCl (1.2 mL,12 mol/L) was added to the solution. Stirring at 80 ℃ for 6 hours, and after the reaction is finished, adding 2mol/L NaOH to adjust the pH of the reaction mixture to 8-9. The aqueous phase was extracted with EA (10 mL. Times.3), the organic layer was washed with brine, na ₂ SO ₄ Drying and concentrating. The residue was purified by column (1% to 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).

Eighth step

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,4.11mmol,50% in H) was added ₂ O), and the reaction was stirred at 80 ℃ for 48H. The mixture was cooled to 25 ℃, diluted with water (3 mL), and extracted with dichloromethane. The pH of the aqueous phase was adjusted to pH by adding 2mol/L NaOH>11. The aqueous phase was extracted with EA (10 mL. Times.3), followed by Na ₂ SO ₄ Dried and concentrated to give 1- (4-fluoro-2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) guanidine 6C (150 mg, crude) as a brown solid.

Ninth step

1- (4-fluoro-2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) guanidine 6C (300 mg,0.98 mmol) and 6- ((dimethylamino) methylene) -1- (2-hydroxyethyl) -7-oxo-4, 5,6, 7-tetrahydro-1H-indazole-3-carboxylic acid ethyl ester 6F were stirred in DMF solution (3.5 mL) at 25 ℃. After stirring at 80 ℃ for 12 hours, the reaction mixture was evaporated under reduced pressure. The residue was purified by column (2% to 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-carboxylate 6D (150 mg, 26.7%) as a pale yellow solid. LCMS (m+h) +=526.

Tenth step

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 ₃ The solution in MeOH (1.5 mL, 7M) was stirred at 80℃in a 10mL tube sealer 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- (4-methylpiperazin-1-yl) phenyl) amino) -1- (2-hydroxyethyl) -4, 5-dihydro-1H-pyrazolo [4, 3-H) as a pale yellow solid]Quinazoline-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

At N ₂ Guanidine carbonate (849.4 mg,4.64 mmol) was added to a solution of (Z) -6- ((dimethylamino) methylene) -1- (2-hydroxyethyl) -7-oxo-4, 5,6, 7-tetrahydro-1H-indazole-3-carboxylic acid ethyl ester 6F (570 mg,1.88 mmol) in DMF (50 ml) under an atmosphere. After stirring at 110℃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% to 50% ea/PE) to give 8-amino-1- (2-hydroxyethyl) -4, 5-dihydro-1H-pyrazolo [4,3-H ] as a pale yellow solid]Quinazoline-3-carboxylic acid ethyl ester 7F_1 (307 mg, 53.8%). ESI-MS (M+H) ⁺ ＝304。

Second step

At N ₂ Next, 8-amino-1- (2-hydroxyethyl) -4, 5-dihydro-1H-pyrazolo [4,3-H]To a solution of quinazoline-3-carboxylic acid ethyl ester 7F_1 (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 (319 mg, 1.00) and isoamyl nitrite (176 mg,1.50 mmol). The reaction was stirred at 80℃for 12 hours. The reaction mixture was diluted with DCM (50 mL) and 20% NH ₃ ·H ₂ O (80 mL), saturated Na ₂ S ₂ O ₃ The aqueous solution (80 mL) and brine (80 mL) were washed, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (2.5% to 10% meoh in DCM) to give 1- (2-hydroxyethyl) -8-iodo-4, 5-dihydro-1H-pyrazolo [4,3-H ] as a yellow solid]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 under N ₂ Potassium nitrate (14 g,138 mmol) and concentrated sulfuric acid (60 ml) were added. The reaction was stirred at 25℃for 2 hours. The reaction mixture was quenched with water (200 mL), extracted with DCM (50 mL. Times.3), and the combined organic layers were extracted with saturated NaHCO ₃ The aqueous solution (100 mL) and brine (100 mL) were washed, dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. The residue was purified by column (0% to 5% ea in PE) to give 1-bromo-2-fluoro-5-nitro-4- (trifluoromethoxy) benzene 7B (30 g, 95%) as a pale yellow oil. ESI-MS (M+H) ⁺ ＝303。

Fourth step

At N ₂ 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) was added iron powder (22.0 g,0.393 mol). The reaction was stirred at 25℃for 3 hours. The reaction mixture was concentrated and diluted with water (100 mL), extracted with EA (30 ml×3), and 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% to 20% ea in PE) to give 5-bromo-4-fluoro-2- (trifluoromethoxy) aniline 7C (21.2 g, 93.8%) as a yellow solid. ¹ HNMR(400MHz,CDCl3)δH 7.00-6.95(m,2H),3.79(brs,2H)。ESI-MS(M+H) ⁺ ＝274.1。

Fifth step

Compound 7D was obtained by a two-step reaction starting from compound 7C according to the synthesis method of example 6.

Sixth step

At N ₂ Down to 1- (2-hydroxy)Ethyl) -8-iodo-4, 5-dihydro-1H-pyrazolo [4,3-H]To a solution of quinazoline-3-carboxylic acid ethyl ester 7F_2 (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 (3.2mg，0.0144mmol)、BINAP(18.0mg，0.0289mmol)，K ₂ CO ₃ (59.6 mg,0.432 mmol). The reaction was stirred at 80℃for 4 hours. The reaction mixture was concentrated and diluted with water (20 mL), extracted with EA (10 ml×3), and 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- (trifluoromethoxy) phenyl) amino) -1- (2-hydroxyethyl) -4, 5-dihydro-1H-pyrazolo [4, 3-H) as a pale yellow solid ]Quinazoline-3-carboxylic acid ethyl ester 7F (75 mg, 16%). ESI-MS (M+H) ⁺ ＝580。

Seventh step

According to the method of example 6, starting from compound 7F (75 mg,0.129 mmol), compound 7 (5.2 mg, 7.3%) was obtained in one step.

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

Compound 8A was obtained in 3 steps starting from compound 8-1 according to the synthesis method of example 6. 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) -4, 5-dihydro-1H-pyrazolo [4, 3-H) at 25 DEG C]Quinazoline-3-carboxylic acid ethyl ester 8A (100 mg,0.138 mmol) to a solution of DCM (5 mL) was added TFA (0.5 mL) and stirred at 25℃for 4 h. Adding saturated NaHCO ₃ The reaction was quenched with aqueous solution (30 mL), extracted with DCM (15 mL. Times.2), the organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered, evaporated under reduced pressure, and the residue was purified by column chromatography (2% to 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

Compound 8C was obtained by one-step reaction starting from compound 8B according to the synthesis method of example 6.

Third step

To 8- ((4-fluoro-5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) -1- (2-hydroxyethyl) -4, 5-dihydro-1H-pyrazolo [4,3-H at 0 ] ]To a solution of fluquinconazole-3-carboxamide 8C (30 mg,0.066 mmol) in DMF (3 mL) was added Cs ₂ CO ₃ (32.6 mg,0.1 mmol) and 2-chloroacetonitrile (6.0 mg, 0.08mmol in 1mL DMF). After stirring at 25℃for 4 hours, the reaction mixture was quenched with water (10 mL). The mixture was extracted with EA (15 ml×2), and 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-methylpiperazin-1-yl) phenyl) amino) -4, 5-dihydro-1H-pyrazolo [4,3-H ] as a pale yellow solid]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 synthesis method of example 8, compound 9 was obtained by 4 steps of reaction starting from compound 8-3 and compound 7D.

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 synthesis method of examples 7 and 8, compound 10 was obtained by starting from compound 8-3 and compound 10-1 through 4 steps of 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

Compound 11 was obtained by 1 step reaction starting from compound 10C according to the synthesis method of example 9.

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 ℃ 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-carboxamide 12E_1 (2.8 g, 93.3%) as a white solid. LCMS (m+h) +=117.

Second step

at-5-0deg.C, N ₂ A suspension of 1-hydroxycyclopropane-1-formylhydrazine 12E_1 (3.0 g,258 mmol) in THF (15 mL) was added drop wise to BH under an atmosphere ₃ THF (258 mL,258mmol, 1M). The reaction mixture was stirred at 60℃for 21 hours. At-5 to 0deg.C at N ₂ Under the protection ofMeOH (50 mL) was added dropwise to the reaction mixture, which was then stirred at 20-25℃for 0.5 hours, and the reaction mixture was concentrated to give a residue. The residue was dissolved in 50mL MeOH and stirred at 80 ℃ for 3 hours. The mixture was cooled to 25 ℃ and evaporated to a residue. The residue was triturated with DCM (30 mL) for 0.5 h and filtered. The filtrate was evaporated to give the crude product 1- (hydrazinomethyl) cyclopropan-1-ol 12e_2 (2.2 g, 75%) as a pale yellow oil. ¹ H NMR(400MHz,CDCl ₃ )d2.91(s,2H),0.83(t,J＝6.0Hz,2H),0.55-0.50(m,2H)。

Third step

Compound 12 was obtained by 4 steps of reaction starting from compound 12e_2 and compound 6f_3 according to the synthesis method of example 9.

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

Compound 13 was obtained by 1 step reaction starting from compound 10C according to the synthesis method of example 9.

¹ H NMR(400MHz,DMSO-d6)δ9.12(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

Compound 14 was obtained by 1 step reaction starting from compound 10C according to the synthesis method of example 9.

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＝9.1,3.0Hz,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

Compound 15 was obtained by 1 step reaction starting from compound 10C according to the synthesis method of example 9.

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

Cs was added to a solution of 16A (193 mg,1 mmol) in DMF (5 mL) at 0deg.C ₂ CO ₃ (650 mg,2 mmol) and (2-bromoethoxy) t-butyldimethylsilane (480 mg,2 mmol). The reaction mixture was stirred overnight at 25℃and quenched with water (10 mL). The mixture was extracted with EA (15 ml×2), and 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 510mg was treated with tris (dimethylamino) methane (2 mL) and the reaction was stirred overnight at 90℃and the volatiles were removed under reduced pressure and the residue was used without further purification.

Third step

According to the synthesis method of example 10, compound 16D was obtained by 1-step reaction starting from compound 16B and compound 15-1.

Fourth step

Methyl 9- (2- ((tert-butyldimethylsilyl) oxy) ethyl) -2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) -6, 9-dihydro-5H-pyrrolo [3,2-H]Methyl quinazoline-7-carboxylate 16D (330 mg,0.5 mmol) was suspended in dioxane (5 mL) and treated with 2N NaOH solution (5 mL,10 mmol) at reflux temperature overnight and H was added ₂ O (50 mL) and acidifying the solution with 2N HCl ph=4. Concentrated under reduced pressure and the residue was purified by column chromatography (DCM: meOH 10:1) to give 9- (2-hydroxyethyl) -2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) -6, 9-dihydro-5H-pyrrolo [3,2-H as a yellow solid]Quinazoline-7-carboxylic acid (240 mg, 90% yield). LC-MS (M+H) +:533.

Fifth step

9- (2-hydroxyethyl) -2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) -6, 9-dihydro-5H-pyrrolo [3,2-H]Quinazoline-7-carboxylic acid (240 mg,0.45 mmol) was dissolved in DMF and HOBt (121 mg,0.9 mmol) and EDCI (172 mg,0.9 mmol) were added to the reaction mixture and stirred for 0.5h. Subsequently, NH is added ₄ OH (4 eq). 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) -2- (trifluoromethoxy) phenyl) amino) -6, 9-dihydro-5H-pyrrolo [3, 2-H) by preparative HPLC]Quinazoline-7-carboxamide, white solid (106 mg, 47% yield).

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

LiHMDS (1M solution in THF, 6.06mL,9.09 mmol) was added dropwise to a solution of cyclopropylmethanol 17A (721.00. Mu.L, 9.09 mmol) in THF (24 mL) at 5 ℃. After 30 min, 4-fluoro-3-nitrobromobenzene (1.11 mL,9.09 mmol) was added quickly and the reaction mixture was allowed to warm to room temperature naturally and stirred overnight. The reaction mixture was quenched with water and extracted with EA, the organic layer was collected 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 3.02g of 4-bromo-1- (cyclopropylmethoxy) -2-nitrobenzene 17B. LCMS (M+H) ⁺ ＝272。

Second step

N ₂ 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), pd under an atmosphere ₂ (dba) ₃ (0.39g，0.43mmol，Aldrich)、Cs ₂ CO ₃ (1.4 g,4.3 mmol) and 1-methylpiperazine (0.43 g,4.3 mmol). After heating overnight at 90 ℃, the reaction was quenched with water, extracted with ethyl acetate (50 ml×3), the organic phase was dried over sodium sulfate, concentrated, and purified by column chromatography (dichloromethane/methanol 20:1) to give 1- (4- (cyclopropylmethoxy) -3-nitrophenyl) -4-methylpiperazine 17C (383 mg,1.32mmol, 61%) as a tan solid. ESI-MS (m+h) +=292.

Third step

1- (4- (cyclopropylmethoxy) -3-nitrophenyl) -4-methylpiperazine 17C (29 mg,1 mmol) was dissolved in a mixed solvent of acetic acid (10 mL) and ethanol (10 mL), zinc powder (650 mg,10 mmol) was added, stirred at 80℃for 3h, filtered over celite, concentrated, and purified by column chromatography (dichloromethane/methanol 20:1) to give 2- (cyclopropylmethoxy) -5- (4-methylpiperazin-1-yl) aniline 17D (185 mg,0.71mmol, 71%). ESI-MS (m+h) +=262.

Fourth step

According to the synthesis method of example 7, starting from compound 17D and 7f_2, the compound 17,2- ((2- (cyclopropylmethoxy) -5- (4-methylpiperazin-1-yl) phenyl) amino) -9- (2-hydroxyethyl) -6, 9-dihydro-5H-pyrrolo [3,2-H ] quinazoline-7-carboxamide is obtained in 2 steps.

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＝4.9Hz,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 CN 101563351B. 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 50mL sealed tube at-78deg.C. After stirring at-78 ℃ for 24 hours, the reaction mixture was cooled to 0 ℃ and then water (20 mL) was added. The mixture was extracted with EA (20 ml×3), and 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% to 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) was in a 50mL sealed tube at 25℃EDCI (70.5 mg,0.347 mmol), DMAP (16.3 mg,0.134 mmol) and O-methyl-hydroxylamine chloride salt (35.3 mg,0.4 mmol). After stirring at 30 ℃ for 2 hours, concentration to remove 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), cyclopropyl alcohol (1.32 g,0.023 mol), K ₂ CO ₃ (9.42 g,0.068 mol) and DMF (10 mL). The reaction was stirred overnight at 25 ℃. By adding H ₂ The reaction mixture was quenched with O (100 mL), extracted with EA (100 mL. Times.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) to give 20A (5.3 g, 90.4%) as a yellow solid.

Compound 14 was obtained by 4 steps of reaction starting from compound 20A according to the synthesis method of example 17.

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 is purchased from SignalChem;

ADP-Glo Kinase Assay is available from Promega;

kinase assay buffer III from SignalChem;

nivo Multi-labelling Analyzer (Perkinelmer).

The experimental method comprises the following steps:

the enzyme, substrate, ATP and inhibitor were diluted using a kinase buffer in the kit. Test compounds were diluted to 1mM with 100% DMSO as the first concentration and then 5-fold diluted to the 8 th concentration, i.e., from 1mM to 0.013. Mu.M, with a row gun. Compound working solutions containing 5% dmso were prepared by diluting each concentration point of the compound 20-fold with 1X kinase buffer, and 1 μl of each concentration gradient working solution of the compound was added to the microplates to set up double wells. To the microplate was added 2. Mu.l PLK1 enzyme (15 ng), 2. Mu.l of a mixture of substrate and ATP (20. Mu.M ATP, 0.2. Mu.g/. Mu.l Casein protein) at a final concentration gradient of 10. Mu.M diluted to 0.13nM and the reaction was allowed to react at 25℃for 60 minutes. After the reaction was completed, 5. Mu.l of ADP-Glo reagent was added to each well, the reaction was continued at 25℃for 40 minutes, 10. Mu.l of the kinase detection reagent was added to each well after the completion of the reaction, and after 30 minutes of the reaction at 25℃the chemiluminescence was read using a Perkinelmer Nivo multi-label analyzer, with an integration time of 0.5 seconds.

Data analysis:

raw data was converted to inhibition ratio, IC, using the equation (Sample-Min)/(Max-Min) ×100% ₅₀ The values of (a) can be obtained by curve fitting with four parameters (log (inhibitor) vs. response-Variable slope mode in GraphPad Prism).

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

Numbering of compounds	PLK1 IC ₅₀ (nM)
6	47.17
7	101.8

8	824.9
9	526
10	9.55
11	8.57
12	9.67
13	18.94
14	50.12
15	14.62
16	6.39
17	9.71

B. In vitro cell proliferation assay

The inhibitory activity of the compounds on tumor cell proliferation was determined using the MTT method. Inoculating HT-29 tumor cells in logarithmic cell growth phase into culture plate, culturing for 24 hr, adding different concentration inhibitors, and culturing at 37deg.C and 5% CO ₂ Culturing for 48 hr, adding 20 μl MTT solution into each well, culturing for 4 hr, dissolving in DMSO, crystallizing, and measuring OD value at 570nm with ELISA ₅₀ 。

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

Numbering of compounds	HT-29 IC ₅₀ (nM)
6	269
10	111
11	232.2
12	75
15	71

Claims

Pyrazoloquinazoline compound as shown in formula I, pharmaceutically acceptable salt thereof, solvate thereof or solvate of pharmaceutically acceptable salt thereof:

wherein R is ¹ Is C ₁ ～C ₄ Alkoxy, substituted by one or more R ^1-1 Substituted C ₁ ～C ₄ Alkoxy or 3-6 membered cycloalkyloxy; r is R ^1-1 Independently halogen or 3-6 membered cycloalkane;

R ² is H, halogen or C ₁ ～C ₄ An alkyl group;

R ³ is 5-10 membered heterocycloalkyl or substituted by one or more R ^3-1 Substituted 5-to 10-membered heterocycloalkyl; in the 5-10 membered heterocycloalkyl, the hetero atom is one or more selected from N, O and S, and the number of the hetero atom is 1-2; r is R ^3-1 Independently C ₁ ～C ₄ An alkyl group;

R ⁴ is cyano, 3-6 membered cycloalkane, substituted with one or more R ^4-1 Substituted 3-6 membered cycloalkanes3-to 6-membered heterocycloalkanes or by one or more R ^4-2 Substituted 3-6 membered heterocycloalkanes; is/are R ^4-3 Substituted C ₁ ～C ₄ An alkyl group; in the 3-6 membered heterocycloalkyl, the hetero atom is one or more selected from N, O and S, and the number of the hetero atom is 1-2; r is R ^4-1 Independently halogen, hydroxy or C ₁ ～C ₄ An alkyl group; r is R ^4-2 Independently halogen or C ₁ ～C ₄ An alkyl group; r is R ^4-3 Independently halogen, cyano or hydroxy;

R ⁵ is H, hydroxy, C ₁ ～C ₄ Alkyl or C ₁ ～C ₄ An alkoxy group.
The pyrazoloquinazoline compound of formula I, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof according to claim 1, wherein R ^1-1 Independently F, cl, br or I, e.g., F;

And/or R ^1-1 Independently a 3-6 membered cycloalkane, said 3-6 membered cycloalkane being a cyclopropane, a cyclobutane or a cyclopentane, for example a cyclopropane or a cyclobutane;

and/or when R ¹ Is C ₁ ～C ₄ In the case of alkoxy, said C ₁ ～C ₄ Alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, for example methoxy;

and/or when R ¹ To be covered by one or more R ^1-1 Substituted C ₁ ～C ₄ In the case of alkoxy, said C ₁ ～C ₄ Alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, for example methoxy;

and/or when R ¹ When the 3-6 membered cycloalkyl oxygen radical is, the 3-6 membered cycloalkoxy radical isCyclopropyloxy, cyclobutyloxy or cyclopentyloxy, for example cyclopropyloxy or cyclobutyloxy;

and/or when R ² When the halogen is halogen, the halogen is F, cl, br or I;

and/or when R ² Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl;

and/or when R ^3-1 Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl;

and/or when R ³ To be covered by one or more R ^3-1 In the case of substituted 5-to 10-membered heterocycloalkyl, the 5-to 10-membered heterocycloalkyl is 6-to 9-membered heterocycloalkyl, for example 6-to 9-membered heterocycloalkyl containing two N, for 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 octahydro-pyrrole [3,4-c ]]A pyrrole group;

and/or when R ^4-1 In the case of halogen, the halogen is F, cl, br or I, such as F;

and/or when R ^4-1 Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl or ethyl;

and/or when R ^4-2 Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, e.g. methyl or ethylA base;

and/or when R ^4-2 In the case of halogen, the halogen is F, cl, br or I, such as F;

and/or when R ^4-3 In the case of halogen, the halogen is F, cl, br or I, such as F;

and/or when R ⁴ In the case of 3-to 6-membered cycloalkanes, the 3-to 6-membered cycloalkanes are cyclopropane, cyclobutane or cyclopentane, for example cyclopropane or cyclobutane;

and/or when R ⁴ To be covered by one or more R ^4-1 When substituted 3-6 membered cycloalkane, the 3-6 membered cycloalkane is cyclopropane, cyclobutane or cyclopentane, for example, cyclopropane or cyclobutane;

and/or when R ⁴ To be covered by one or more R ^4-2 When the 3-6 membered heterocyclic alkane is substituted, the 3-6 membered heterocyclic alkane is 4 membered heterocyclic alkane containing one N or O or 5 membered heterocyclic alkane containing one N or O;

and/or when R ⁵ Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl is methyl, ethyl, isopropyl or tert-butyl, for example methyl or ethyl;

and/or when R ⁵ Is C ₁ ～C ₄ In the case of alkoxy, said C ₁ ～C ₄ Alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, for example methoxy.
The pyrazoloquinazoline compound of formula I, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof according to claim 1, wherein R ¹ Is that For example

And/or R ³ Is that

And/or R ³ Is that

And/or R ³ Is that

And/or when R ⁴ To be covered by one or more R ^4-1 Substituted 3-to 6-membered cycloalkanes, said substituted with one or more R ^4-1 Substituted 3-6 membered cycloalkanes as

And/or when R ⁴ In the case of 3-6 membered heterocyclic alkane, the 3-6 membered heterocyclic alkane is

And/or when R ⁴ To be covered by one or more R ^4-2 In the case of substituted 3-to 6-membered heterocycloalkanes, said substituted 3-to 6-membered heterocycloalkanes are substituted by one or more R ^4-2 Substituted 3-to 6-membered heterocycloalkanes

And/or when R ⁴ To be covered by one or more R ^4-3 Substituted C ₁ ～C ₄ In the case of alkyl groups, said groups are substituted by one or more R ^4-3 Substituted C ₁ ～C ₄ Alkyl is
The pyrazoloquinazoline compound of formula I, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof according to claim 1, wherein R ⁴ Is cyano, covered by one or more R ^4-1 Substituted 3-6 membered cycloalkanes, 3-6 membered heterocycloalkanes, substituted by one or more R ^4-2 Substituted 3-6 membered heterocycloalkyl, or by one or more R ^4-3 Substituted C ₁ ～C ₄ An alkyl group;

and/or R ^4-1 Halogen or hydroxy;

and/or R ^4-2 Is C ₁ ～C ₄ An alkyl group;

and/or R ^4-3 Is hydroxyl;

and/or R ⁵ Is H, hydroxy or C ₁ ～C ₄ An alkoxy group.
The pyrazoloquinazoline compound of formula I, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof according to claim 1, wherein R ⁴ Is cyano, covered by one or more R ^4-1 Substituted 3-6 membered cycloalkanes, 3-6 membered heterocycloalkanes, substituted by one or more R ^4-2 Substituted 3-6 membered heterocycloalkyl, or by one or more R ^4-3 Substituted C ₁ ～C ₄ An alkyl group;

R ^4-1 halogen or hydroxy;

R ^4-2 is C ₁ ～C ₄ An alkyl group;

R ^4-3 is hydroxyl;

R ⁵ is H, hydroxy or C ₁ ～C ₄ An alkoxy group.
The pyrazoloquinazoline compound of formula I, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof according to claim 5 wherein R ⁴ Selected from the group consisting of: cyano group,
The pyrazoloquinazoline compound of formula I, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof according to claim 1, having the formula:
the pyrazoloquinazoline compound of formula I, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof according to claim 7 wherein R ¹ To be covered by one or more R ^1-1 Substituted C ₁ ～C ₄ An alkoxy group.
The pyrazoloquinazoline compound of formula I, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof according to claim 7 wherein R ² Is H or halogen.
The pyrazoloquinazoline compound of formula I, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof according to claim 7 wherein R ⁴ Selected from the group consisting of: cyano group,
A pyrazoloquinazoline compound, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof, wherein the pyrazoloquinazoline compound is selected from any one of the following:
A process for preparing pyrazoloquinazoline compound, its pharmaceutically acceptable salt, its solvate or its solvate of pharmaceutically acceptable salt as shown in formula I includes such steps as preparing 1g of compound, ligand reagent and R in organic solvent under the action of Pd catalyst in alkaline reagent ³ Carrying out coupling reaction on H to obtain a compound I;

wherein R is ¹ 、R ² 、R ³ 、R ⁴ Or R is ⁵ Is as defined in claims 1 to 6; x is halogen.
The method of claim 12, wherein the palladium catalyst is dibenzylidene acetone dipalladium;

and/or the organic solvent is tetradioxane;

and/or, the alkali reagent is cesium carbonate;

and/or the ligand reagent is 4, 5-bis (diphenylphosphine) -9, 9-dimethylxanthene.
Use of a pyrazoloquinazoline compound of formula I, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 11 in the manufacture of a PLK1 inhibitor; the inhibitor is preferably an inhibitor for use in vitro.
A pharmaceutical composition comprising a pyrazoloquinazoline compound of formula I according to claims 1 to 11, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable adjuvant.
Use of a pyrazoloquinazoline compound of any one of claims 1 to 11 as shown in formula I, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof in the manufacture of a medicament;

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

wherein R is ¹ To be covered by one or more R ^1-1 Substituted C ₁ ～C ₄ An alkoxy group; r is R ^1-1 Independently halogen;

R ² is H, halogen or C ₁ ～C ₄ An alkyl group;

R ^3’ is covered by one R ^3-1 Substituted or unsubstituted 7-to 9-membered heterocycloalkyl; the 7-9 membered heterocycloalkyl is 7-9 membered hetero spirocycloalkyl or 7-9 membered hetero bridged cycloalkyl; the hetero atoms are selected from one or more of N, O and S, and the number of the hetero atoms is 1-2; r is R ^3-1 Is C ₁ ～C ₄ An alkyl group;

R ^4’ independently is covered by one R ^4-3 Substituted C ₁ ～C ₄ An alkyl group; r is R ^4-3 Is hydroxy or halogen;

R ⁵ is H, hydroxy, C ₁ ～C ₄ Alkyl or C ₁ ～C ₄ An alkoxy group.
The process for producing pyrazoloquinazoline compound of the formula II, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof according to claim 17, wherein R ^1-1 F, cl, br or I, e.g., F;

and/or when R ¹ To be covered by one or more R ^1-1 Substituted C ₁ ～C ₄ In the case of alkoxy, said C ₁ ～C ₄ Alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, for example methoxy;

and/or when R ² Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl;

and/or R ^3-1 Methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl;

and/or when R ^4-3 When the halogen is halogen, the halogen is F, cl, br or I;

and/or when R ^4’ Independently is covered by one R ^4-3 Substituted C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl;

and/or when R ⁵ Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl is methyl, ethyl, isopropyl or tert-butyl, for example methyl or ethyl;

and/or when R ⁵ Is C ₁ ～C ₄ In the case of alkoxy, said C ₁ ～C ₄ Alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, for example methoxy.
The pyrazoloquinazoline compound of the formula II, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof according to claim 17, wherein R ¹ Is thatFor example

And/or when R ^3’ In the case of a 7-to 9-membered heterospirocycloalkyl group, the 7-to 9-membered heterospirocycloalkyl group is a 7-to 9-membered heterospirocycloalkyl group having two nitrogen atoms, for example

And/or when R ^3’ Is covered by one R ^3-1 In the case of substituted 7-to 9-membered heterospirocycloalkyl, said residue is substituted with one R ^3-1 Substituted or unsubstituted 7-to 9-membered heterospirocycloalkyl being substituted by one R containing two nitrogen atoms ^3-1 Substituted 7-to 9-membered heterospirocycloalkyl groups, e.g.

And/or when R ^3’ In the case of a 7-to 9-membered heterobridged cycloalkyl group, the 7-to 9-membered heterobridged cycloalkyl group is a 7-to 9-membered heterobridged cycloalkyl group containing two N groups, for example

And/or when R ^3’ Is covered by one R ^3-1 In the case of substituted 7-to 9-membered heterobridged cycloalkyl, said substituted is represented by one R ^3-1 Substituted 7-to 9-membered heterobridged cycloalkyl radicals being substituted by one R, containing two nitrogen atoms ^3-1 Substituted 7-to 9-membered heterobridged cycloalkyl radicals, e.g.

And/or R ⁴ is-CH ₂ OH。
The pyrazoloquinazoline compound of formula II, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof according to claim 17, wherein the pyrazoloquinazoline compound of formula II, a pharmaceutically acceptable salt thereof, a solvate of a pharmaceutically acceptable salt thereof, or a solvate of a pharmaceutically acceptable salt thereof is as follows:
A process for preparing pyrazoloquinazoline compound, its pharmaceutically acceptable salt, its solvate or its solvate of pharmaceutically acceptable salt as shown in formula II includes such steps as preparing 1g of compound, ligand reagent and R under the action of Pd catalyst in organic solvent and alkali reagent ³ Carrying out coupling reaction on H to obtain a compound II;

wherein R is ¹ 、R ² 、R ^3’ 、R ^4’ Or R is ⁵ Is as defined in claims 12 to 15; x is halogen.
The method of claim 21, wherein the palladium catalyst is dibenzylidene acetone dipalladium;

and/or the organic solvent is tetradioxane;

and/or, the 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 is ¹ Is C ₁ ～C ₄ Alkoxy, substituted by one or more R ^1-1 Substituted C ₁ ～C ₄ Alkoxy, R ^1- ¹ Independently halogen or 3-6 membered cycloalkane;

R ² is H, halogen or C ₁ ～C ₄ An alkyl group;

R ³ is 5-10 membered heterocycloalkyl or substituted by one or more R ^3-1 Substituted 5-to 10-membered heterocycloalkyl; in the 5-10 membered heterocycloalkyl, the hetero atom is one or more selected from N, O and S, and the number of the hetero atom is 1-2; r is R ^3-1 Independently C ₁ ～C ₄ An alkyl group;

R ⁴ is cyano, 3-6 membered cycloalkane, substituted with one or more R ^4-1 Substituted 3-6Of a cycloalkane, a 3-to 6-membered heterocycloalkyl or by one or more R ^4-2 Substituted 3-6 membered heterocycloalkanes; is/are R ^4-3 Substituted C ₁ ～C ₄ An alkyl group; in the 3-6 membered heterocycloalkyl, the hetero atom is one or more selected from N, O and S, and the number of the hetero atom is 1-2; r is R ^4-1 Independently halogen, hydroxy or C ₁ ～C ₄ An alkyl group; r is R ^4-2 Independently halogen or C ₁ ～C ₄ An alkyl group; r is R ^4-3 Independently halogen, cyano or hydroxy;

R ⁵ is H, hydroxy C ₁ ～C ₄ Alkyl or C ₁ ～C ₄ An alkoxy group.
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 Independently F, cl, br or I, e.g., F;

and/or R ^1-1 Independently 3-6 membered cycloalkane is cyclopropane, cyclobutane or cyclopentane, for example cyclopropane or cyclobutane;

and/or when R ¹ Is C ₁ ～C ₄ In the case of alkoxy, said C ₁ ～C ₄ Alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, for example methoxy;

and/or when R ¹ To be covered by one or more R ^1-1 Substituted C ₁ ～C ₄ In the case of alkoxy, said C ₁ ～C ₄ Alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, for example methoxy;

And/or when R ² When the halogen is halogen, the halogen is F, cl, br or I;

and/or the number of the groups of groups,when R is ² Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl;

and/or when R ^3-1 Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl;

and/or when R ³ To be covered by one or more R ^3-1 In the case of substituted 5-to 10-membered heterocycloalkyl, the 5-to 10-membered heterocycloalkyl is 6-to 9-membered heterocycloalkyl, for example 6-to 9-membered heterocycloalkyl containing two N, for 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 octahydro-pyrrole [3,4-c ]]A pyrrole group;

and/or when R ^4-1 In the case of halogen, the halogen is F, cl, br or I, such as F;

and/or when R ^4-1 Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl or ethyl;

And/or when R ^4-2 Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl or ethyl;

and/or when R ^4-2 In the case of halogen, the halogen is F, cl, br or I, such as F;

and/or when R ^4-3 When halogen is halogen, the halogen is F, cl, br or I,such as F;

and/or when R ⁴ In the case of 3-to 6-membered cycloalkanes, the 3-to 6-membered cycloalkanes are cyclopropane, cyclobutane or cyclopentane, for example cyclopropane or cyclobutane;

and/or when R ⁴ To be covered by one or more R ^4-1 When substituted 3-6 membered cycloalkane, the 3-6 membered cycloalkane is cyclopropane, cyclobutane or cyclopentane, for example, cyclopropane or cyclobutane;

and/or when R ⁴ To be covered by one or more R ^4-2 When the 3-6 membered heterocyclic alkane is substituted, the 3-6 membered heterocyclic alkane is 4 membered heterocyclic alkane containing one N or O or 5 membered heterocyclic alkane containing one N or O;

and/or when R ⁵ Is C ₁ ～C ₄ In the case of alkyl, said C ₁ ～C ₄ Alkyl is methyl, ethyl, isopropyl or tert-butyl, for example methyl or ethyl;

and/or when R ⁵ Is C ₁ ～C ₄ In the case of alkoxy, said C ₁ ～C ₄ Alkoxy is methoxy, ethoxy, isopropoxy or tert-butoxy, for example methoxy.
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 Is thatFor example

And/or R ³ Is that

And/or R ³ Is that

And/or R ³ Is that

And/or when R ⁴ To be covered by one or more R ^4-1 Substituted 3-to 6-membered cycloalkanes, said substituted with one or more R ^4-1 Substituted 3-6 membered cycloalkanes as

And/or when R ⁴ In the case of 3-6 membered heterocyclic alkane, the 3-6 membered heterocyclic alkane is

And/or when R ⁴ To be covered by one or more R ^4-2 In the case of substituted 3-to 6-membered heterocycloalkanes, said substituted 3-to 6-membered heterocycloalkanes are substituted by one or more R ^4-2 Substituted 3-to 6-membered heterocycloalkanes

And/or when R ⁴ To be covered by one or more R ^4-3 Substituted C ₁ ～C ₄ In the case of alkyl groups, said groups are substituted by one or more R ^4-3 Substituted C ₁ ～C ₄ Alkyl is
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 ⁴ Is cyano, covered by one or more R ^4-1 Substituted 3-to 6-membered cycloalkanes, 3-to 6-membered heterocycloalkanes or by one or more R ^4-2 Substituted 3-6 membered heterocycloalkyl, or by one or more R ^4-3 Substituted C ₁ ～C ₄ An alkyl group;

and/or R ^4-1 Halogen or hydroxy;

and/or R ^4-2 Is C ₁ ～C ₄ An alkyl group;

and/or R ^4-3 Is hydroxyl;

and/or R ⁵ Is H, hydroxy or C ₁ ～C ₄ An alkoxy group.
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 ⁴ Is cyano, covered by one or more R ^4-1 Substituted 3-6 membered cycloalkanes, 3-6 membered heterocycloalkanes, substituted by one or more R ^4-2 Substituted 3-6 membered heterocycloalkyl, or by one or more R ^4-3 Substituted C ₁ ～C ₄ An alkyl group;

R ^4-1 halogen or hydroxy;

R ^4-2 is C ₁ ～C ₄ An alkyl group;

R ^4-3 is hydroxyl;

R ⁵ is H, hydroxy or C ₁ ～C ₄ An alkoxy group.
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 a compound of 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 inhibitor is preferably an inhibitor for use in vitro.
A pharmaceutical composition comprising a compound of any one of claims 1-28, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof, and a pharmaceutical excipient.
Use of 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 in the manufacture of a medicament;

the medicine is used 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 of any one of claims 1-28, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof, and one or more chemotherapeutic agents for simultaneous, separate or sequential use with the compound, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof.