WO2019233458A1 - Vegfr inhibitor, preparation method therefor and use thereof - Google Patents

Abstract

The present invention relates to a quinoline or quinazoline derivative having a structure of formula (I), a pharmaceutical composition comprising the compound of formula (I), and use of the compound in the preparation of a medicament for preventing or treating angiogenesis-associated diseases, in particular for preventing or treating protein tyrosine kinase-associated tumors. Each substituent in formula (I) has the same definition as that in the description.

Description

VEGFR inhibitor and preparation method and application thereof Technical field

The present application belongs to the technical field of medicine, and particularly relates to quinoline or quinazoline derivatives and uses thereof for preparing a medicament for treating malignant tumor diseases.

This application claims the priority of Chinese patent CN201810597224.3 (filed on June 8, 2018, the invention name VEGFR inhibitor, and its preparation method and application).

Background technique

Receptor tyrosine kinases are a class of enzymes that span cell membranes. They have extracellular binding regions, transmembrane structural regions, and intracellular parts that bind growth factors. Residues phosphorylate and affect cell proliferation.

Vascular endothelial growth factor (VEGF), also known as vascular permeability factor (VPF), is a highly specific vascular endothelial cell growth factor, and vascular endothelial growth factor receptors (including VEGFR-1, VEGFR-2, VEGFR- 3) It specifically binds and activates receptor tyrosine kinases to exert vascular regulatory effects, and promotes increased vascular permeability, extracellular matrix degeneration, vascular endothelial cell migration, proliferation, and angiogenesis.

Normal angiogenesis plays an important role in a variety of processes, including embryonic development, wound healing, etc .; bad or pathological angiogenesis is associated with disease states, including diabetic retinopathy, psoriasis, cancer, rheumatoid arthritis, Atherosclerosis. Tumor angiogenesis, the formation of new blood vessels, and their permeability are mainly regulated by (tumor-derived) vascular endothelial cell growth factor (VEGF), which acts through at least two different receptors: VEGFR-1 and VEGFR-2 The receptor is highly specific for vascular endothelial cells (Endocr. Rev. 1992, 13, 18; FASEB J. 1999, 13, 9). VEGFR-1 and VEGFR-2 are mainly distributed on the surface of tumor vascular endothelial cells and regulate tumor angiogenesis; VEGFR-3 is mainly distributed on the surface of lymphatic endothelium and regulate tumor angiogenesis. Most human tumors express VEGF and its receptor at high levels, which has led to the hypothesis that VEGF released by tumor cells stimulates capillary growth and tumor endothelial proliferation in a paracrine manner, and promotes tumor growth by increasing blood supply.

The role of VEGF as a tumor angiogenic factor in vivo is demonstrated by the inhibition of VEGF expression or VEGF activity by VEGF antibodies, VEGFR-2 negative mutants, and VEGF antisense RNA, which can make glioma cell lines or other tumor cell lines grow in vivo Slow down. Several major mechanisms play an important role in anti-tumor angiogenesis-inhibiting activity: inhibition of angiogenesis, especially capillaries, the balance between cell death and proliferation makes the tumor without net growth; due to the lack of blood inflow and outflow from the tumor, Inhibit the metastasis of tumor cells; inhibit the proliferation of endothelial cells and avoid the effect of endothelial cells that arrange blood vessels on the paracrine growth-stimulation of surrounding tissues.

Currently a series of patent applications for VEGFR inhibitors and their applications in vascular related diseases have been published, including WO2016091165, WO2017177962, CN103382206, etc., but there is still a need to develop new novel VEGFR inhibitors with better drug efficacy.

Summary of the Invention

The object of the present invention is to provide a class of quinoline or quinazoline VEGFR inhibitors having excellent activity.

Another object of the present invention is to provide the use of the quinoline or quinazoline VEGFR inhibitor in the preparation of a medicament for preventing or treating angiogenesis-related diseases, in particular to prevent or treat tumor diseases related to protein tyrosine kinase .

To achieve the objective of the present invention, the technical solution of the present invention is as follows:

The compound represented by the following formula (I), its stereoisomers, pharmaceutically acceptable salts or esters or solvates according to the present invention:

R ^{1 is} selected from C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, nitro, -NR ⁸ R ⁹ ,- NR ⁸ COR ⁷ , -COR ⁷ , -SO ₂ R ⁷ , -SOR ⁷ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₈ alkoxy or 4-10 membered heterocyclic The C ₃ -C ₆ cycloalkyl or 4- to 10-membered heterocyclic group may be substituted by an amino group, a hydroxyl group, a-(CH ₂ ) _n CN, a carboxyl group, or a C ₁ -C ₄ alkyl group. Or C ₁ -C ₄ alkoxy.

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

R ^{2 is} selected from H, deuterium or halogen;

X is selected from N or CH;

Y is selected from O or CR ³ R ⁴ ;

R ³ and R ⁴ are each independently selected from H, deuterium, halogen, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or C ₁ -C ₈ alkoxy;

R ^{5 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

R ^{6 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

R ^{7 is} selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, -NR ⁸ R ⁹ or C ₃ -C ₆ heterocyclyl. The C ₃ -C ₆ heterocyclyl may be Further substituted with hydroxy, carboxy or C ₁ -C ₄ alkyl;

R ⁸ and R ⁹ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl.

The embodiment of the present invention is characterized by:

R ¹ is C ₁ -C ₄ alkyl, optionally further substituted with one or more substituents selected from deuterium, hydroxyl, methyl, ethyl, cyclopropyl, or a 4-6 membered heterocyclic group, said The cyclopropyl or 4- to 6-membered heterocyclic group may be further substituted with an amino group, a hydroxyl group, a carboxyl group, a C _{1 to} C ₄ alkyl group, or a C _{1 to} C ₄ alkoxy group. Is selected from pyrrolidinyl, morpholinyl, piperazinyl or piperidinyl.

R ^{2 is} selected from H, deuterium or halogen;

X is CH;

Y is CH ₂ ;

R ³ and R ⁴ are each independently selected from H, deuterium, halogen, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or C ₁ -C ₈ alkoxy;

R ^{5 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

R ^{6 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl.

An embodiment of the present invention is a compound described by the general formula (II), a stereoisomer thereof, a pharmaceutically acceptable salt or ester, or a solvate:

R ¹ is C ₁ -C ₈ alkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, -COR ⁷ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl Or a 4-6 membered heterocyclyl substituent, the C ₃ -C ₆ cycloalkyl or 4-6 membered heterocyclyl may be further substituted with amino, hydroxyl,-(CH ₂ ) _n CN, carboxyl, C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy;

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

R ^{7 is} selected from the group consisting of H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or C ₃ -C ₆ heterocyclyl. The C ₃ -C ₆ heterocyclyl may be further substituted by hydroxy, carboxy or C ₁ -C ₄ alkyl.

The embodiment of the present invention is characterized by:

R ¹ is C ₁ -C ₄ alkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, C ₁ -C ₂ alkyl, C ₃ -C ₆ cycloalkyl, or 4-6 membered heterocyclic group substituents, the C ₃ -C ₆ cycloalkyl or 4-6 membered heterocyclic group may be further substituted by amino, _{hydroxy, - (CH 2) n CN} , carboxy, C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy substituted; preferably, R ¹ is methyl, ethyl, which is further substituted with a hydroxy, methyl, cyclopropyl or pyrrolidinyl substituent, said Cyclopropyl or pyrrolidinyl may be further substituted with amino, hydroxyl,-(CH ₂ ) _n CN, carboxyl, methyl or methoxy;

n is selected from 0, 1 or 2, preferably n is 0 or 1.

The embodiment of the present invention is characterized by:

R ¹ is C ₁ -C ₄ alkyl, optionally further substituted with one or more substituents selected from deuterium, hydroxyl, methyl, ethyl, cyclopropyl, or a 4-6 membered heterocyclic group, said The cyclopropyl or 4- to 6-membered heterocyclic group may be further substituted with an amino group, a hydroxyl group, a carboxyl group, a C _{1 to} C ₄ alkyl group, or a C _{1 to} C ₄ alkoxy group. Is selected from pyrrolidinyl, morpholinyl, piperazinyl or piperidinyl.

In a preferred embodiment of the present invention, the compound is selected from:

Another aspect of the present invention provides a method for preparing a compound of formula (I) or a pharmaceutically acceptable salt or ester or solvate thereof, comprising the following steps:

Among them, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , X, Y are as defined for the compound of formula (I).

The compound of formula (I) or a pharmaceutically acceptable salt or ester or solvate thereof according to the present invention is a novel VEGFR inhibitor, and thus can be used to prepare a medicament for preventing or treating angiogenesis-related diseases, especially for preventing or treating proteins. Tyrosine kinase-associated malignancies.

As a further preferred embodiment, the tumor is selected from the group consisting of ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, non-cancer Hodgkin's lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer, kidney cancer, anaplastic large cell lymphoma, acute myeloid leukemia, multiple bone marrow Tumor, melanoma or mesothelioma, soft tissue sarcoma.

Another aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the aforementioned compound of formula (I) or a pharmaceutically acceptable salt or ester or solvate thereof as an active ingredient and a pharmaceutically acceptable carrier.

Another aspect of the present invention provides the application of the aforementioned pharmaceutical composition in the preparation of a medicament for preventing or treating malignant tumors.

Unless stated to the contrary, the following terms used in the specification and claims have the following meanings.

In the present invention, "C ₁ -C ₈ alkyl" refers to a straight-chain alkyl group and a branched alkyl group including 1 to 8 carbon atoms. The alkyl group refers to a saturated aliphatic hydrocarbon group, such as methyl, ethyl, n- Propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2 -Dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-tris Methylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl Methyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4 -Methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl , 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2- Dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl 2-ethylpentyl, 2-methyl-3-ethylpentyl, or various branched isomers thereof, and the like.

In the present invention, "cycloalkyl" refers to a saturated monocyclic hydrocarbon substituent, and "C ₃ -C ₈ cycloalkyl" refers to a monocyclic cycloalkyl including 3 to 8 carbon atoms, for example, Restrictive examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.

In the present invention, "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein one or more ring atoms are selected from nitrogen, oxygen, or S (O) r (where r is an integer of 0, Heteroatoms of 1, 2), but excluding the ring part of -OO-, -OS- or -SS-, the remaining ring atoms are carbon. "4-10 membered heterocyclyl" refers to a cyclic group containing 4 to 10 ring atoms. Non-limiting examples of monocyclic heterocyclyl include azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like. Polycyclic heterocyclic groups include spiro, fused and bridged heterocyclic groups.

In the present invention, "alkoxy" means -O- (alkyl), wherein alkyl is as defined above. "C ₁ -C ₈ alkoxy" refers to an alkyl group having 1 to 8 carbon atoms, non-limiting embodiments include methoxy, ethoxy, propoxy, butoxy and the like.

"Halogen" means fluorine, chlorine, bromine or iodine.

"Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or a physiologically pharmaceutically acceptable salt or prodrug thereof with other chemical components, as well as other components such as physiologically pharmaceutically acceptable carriers and excipients.形 剂。 Shape agent. The purpose of the pharmaceutical composition is to promote the administration to the organism, which is beneficial to the absorption of the active ingredient and then exerts the biological activity.

In the preparation steps of the present invention, the abbreviations of the reagents used represent:

DCM: Dichloromethane

DIEA N, N-diisopropylethylamine

MTBE, methyl tert-butyl ether

EA, ethyl acetate

PE, petroleum ether

THF: Tetrahydrofuran

TFAA Trifluoroacetic anhydride

EtOH Ethanol

BnBr benzyl bromide

DMAP, 4-dimethylaminopyridine

TFA Trifluoroacetic acid

MsCl, p-toluenesulfonyl chloride

DMAC: Dimethylacetamide

TBDPSCl tert-butyldiphenylchlorosilane

TBAB: Tetrabutylammonium bromide

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 Nuclear magnetic resonance proton spectrum of the compound of Example 1;

Figure 2 Nuclear magnetic resonance proton spectrum of the compound of Example 2;

Figure 3 Nuclear magnetic resonance proton spectrum of the compound of Example 3;

Detailed ways

The present invention is described below with reference to specific examples. Those skilled in the art can understand that these examples are only used to illustrate the present invention, and they do not limit the scope of the present invention in any way.

Unless otherwise specified, the experimental methods in the following examples are conventional methods. Unless otherwise specified, the medicinal materials, reagent materials, etc. used in the following examples are all commercially available products.

Example 1

1-(((9-((4-fluoro-2-methyl-1-hydroindol-5-yl) oxy) -1,2-dihydrofuro [3,2-f] quinoline- 4-yl) oxy) methyl) -cyclopropane-1-aminohydrochloride

Step 1: Synthesis of 2,3-dihydrobenzofuran-7-formyl chloride

In an ice bath, 2,3-dihydrobenzofuran-7-carboxylic acid (13.65 g, 83.15 mmol) and SOCl ₂ (70 ml) were sequentially added to the single-necked flask, and the reaction was terminated at 60 ° C. for 1 h. After reducing to room temperature, it was concentrated under reduced pressure to obtain an off-white solid. Without further purification, it was directly used in the next reaction.

Step 2: Synthesis of N-methoxy-N-methyl-2,3-dihydrobenzofuran-7-formamide

At room temperature, add 2,3-dihydrobenzofuran-7-formyl chloride (15.18 g, 83.13 mmol) to a single-necked flask, dissolve it in DCM (150 ml), add DIEA (30 ml) dropwise under ice bath, and add N, O-dimethylhydroxylamine hydrochloride (9.73 g, 99.75 mmol), the reaction was terminated at room temperature for 1 h. Extracted with EA, washed twice with saturated brine, dried the organic phase over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (EA / PE system) to obtain 16.37 g of a yellow solid with a yield of 95% in two steps. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 7.30 (dd, J = 7.3, 0.9 Hz, 1H), 7.09 (d, J = 7.5 Hz, 1H), 6.86 (t, J = 7.5 Hz, 1H) , 4.56 (t, J = 8.7 Hz, 2H), 3.55 (s, 3H), 3.20 (t, J = 8.8 Hz, 2H), 3.18 (s, 3H).

Step 3: Synthesis of 1- (2,3-dihydrobenzofuran-7-yl) ethanone

At room temperature, N-methoxy-N-methyl-2,3-dihydrobenzofuran-7-carboxamide (16.37 g, 78.99 mmol), THF (450 ml), and N ₂ were added to the three-necked flask in order. Three times, a solution of methyl magnesium bromide in THF (170.00 ml, 170.00 mmol) was added dropwise under an ice bath, and the reaction was terminated at room temperature for 1 h after the dropwise addition. The reaction system was moved to an ice bath, and the pH was adjusted to 3 to 4 with 1N hydrochloric acid. It was then extracted with EA, washed twice with saturated brine, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (EA / PE system) to obtain 11.11 g of an off-white solid with a yield of 85%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.53 (d, J = 7.9 Hz, 1 H), 7.46 (dd, J = 7.2, 1.1 Hz, 1 H), 6.91 (t, J = 7.5 Hz, 1 H) , 4.68 (t, J = 8.8 Hz, 2H), 3.23 (t, J = 8.8 Hz, 2H), 2.52 (s, 3H).

Step 4: Synthesis of 1- (5-nitro-2,3-dihydrobenzofuran-7-yl) ethanone

Add H ₂ SO ₄ (50 ml) to a three-necked flask under ice bath, and add 1- (2,3-dihydrobenzofuran-7-yl) ethanone (11.10 g, 68.44 mmol) and KNO in portions with stirring. ₃ (11.76 g, 116.32 mmol), the reaction was terminated at 0 ° C for 1.5 h. The reaction solution was slowly poured into ice water and stirred for 10 min. The liquid was extracted with EA, and the organic phase was washed once with a saturated sodium bicarbonate solution and a saturated saline solution. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and column chromatography. Purification (EA / PE system) gave 12.20 g of yellow solid with a yield of 86%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.39 (d, J = 2.2 Hz, 1 H), 8.33-8.29 (m, 1 H), 4.90 (t, J = 8.8 Hz, 2 H), 3.35 (t, J = 8.9Hz, 2H), 2.58 (s, 3H).

Step 5: Synthesis of (5-nitro-2,3-dihydrobenzofuran-7-yl) acetate

TFAA (200ml) was added to a single-necked flask, and H ₂ O ₂ (50ml) was added dropwise at -10 ° C. After the drop was completed, 1- (5-nitro-2,3-dihydrogen) was added dropwise after stirring at this temperature for 20min. A solution of benzofuran-7-yl) ethanone (19.00 g, 91.70 mmol) in DCM (45 ml) was added, and the reaction was terminated at room temperature for 3 h. Extracted with EA, washed twice with saturated brine, dried the organic phase over anhydrous sodium sulfate, filtered, and concentrated to give a yellow solid, which was directly used in the next reaction without further purification.

Step 6: Synthesis of 5-nitro-2,3-dihydrobenzofuran-7-ol

(5-nitro-2,3-dihydrobenzofuran-7-yl) acetate (20.47g, 91.71mmol) and EtOH (250ml) were added to a single-necked bottle in turn at room temperature, and 40 drops were added in an ice bath. % NaOH solution (25ml), and the reaction was terminated at room temperature for 0.5h after dropping. Adjust the pH to 3 ~ 4 with 2N hydrochloric acid in an ice bath, extract with EA, and wash twice with saturated brine. The organic phase is dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (EA / PE system) to obtain a yellow solid 15.00g, 90% yield in two steps. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 7.70-7.64 (m, 1H), 7.54 (d, J = 2.3Hz, 1H), 4.70 (t, J = 8.9Hz, 2H), 3.26 (t, J = 8.8Hz, 2H). MS (ESI) m / z: 180.0 [MH] ^- .

Step 7: Synthesis of 7-benzyloxy-5-nitro-2,3-dihydrobenzofuran

At room temperature, 5-nitro-7-hydroxy-2,3-dihydrobenzofuran (15.0 g, 82.80 mmol), DMF (300 ml), K ₂ CO ₃ (171.66 g, 1242.02 mmol) were sequentially added to the single-necked flask. ), BnBr (17.00g, 99.40mmol), the reaction was terminated at 80 ° C for 0.5h. Extracted with EA, washed twice with saturated brine, dried the organic phase over anhydrous sodium sulfate, filtered, concentrated, and then slurried with PE (250ml) at room temperature for 30min, and filtered with suction to obtain 19.42g of yellow solid with a yield of 87%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 7.87–7.83 (m, 1H), 7.80 (d, J = 2.0 Hz, 1H), 7.47–7.33 (m, 5H), 5.24 (s, 2H), 4.74 (t, J = 8.9 Hz, 2H), 3.30 (t, J = 8.9 Hz, 2H).

Step 8: Synthesis of 7-benzyloxy-2,3-dihydrobenzofuran-5-amine

At room temperature, 7-benzyloxy-5-nitro-2,3-dihydrobenzofuran (19.00 g, 70.04 mmol), EtOH (450 ml), H ₂ O (112 ml), NH were sequentially added to the single-necked flask. ₄ Cl (15.69 g, 293.33 mmol) was heated to 80 ° C, Fe powder (30.00 g, 537.20 mmol) was added, and the reaction was terminated at this temperature for 1.5 h. After suction filtration, H ₂ O was added to the filtrate and extracted with EA three times. The organic phases were combined and dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (EA / PE system) to obtain 14.71 g of a black solid with a yield of 87%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 7.42–7.29 (m, 5H), 6.14 (s, 1H), 6.11 (s, 1H), 5.00 (s, 2H), 4.37 (t, J = 8.6 Hz, 2H), 3.01 (t, J = 8.5Hz, 2H). MS (ESI) m / z: 242.2 [M + H] ⁺ .

Step 9: Synthesis of 5- (ethoxymethylene) -2,2-dimethyl-1,3-dioxane-4,6-dione

At room temperature, cyclopropylmalonate (8.36 g, 58.00 mmol), triethyl orthoformate (29.37 ml, 176.58 mmol) were sequentially added to the single-necked flask, and the reaction was terminated at 80 ° C for 1 h. Without further processing, it was directly used in the next reaction.

Step 10: 5-((7-benzyloxy-2,3-dihydrobenzofuran-5-ylamino) methylene) -2,2-dimethyl-1,3-dioxane-4 Synthesis of 1,6-dione

At room temperature, 7-benzyloxy-2,3-dihydrobenzofuran-5-amine (13.99g, 57.98mmol), 5- (ethoxymethylene) -2,2 -Dimethyl-1,3-dioxane-4,6-dione (11.61 g, 57.99 mmol), isopropanol (350 ml), reaction was terminated at 80 ° C for 0.5 h. The temperature was lowered to room temperature, and 19.05 g of a dark yellow solid was obtained by suction filtration, and the yield in two steps was 83%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.50 (d, J = 16.0 Hz, 1H), 7.46–7.32 (m, 5H), 7.26 (d, J = 1.1 Hz, 1H), 7.08 (s, 1H), 5.15 (s, 2H), 4.56 (t, J = 8.8Hz, 2H), 3.19 (t, J = 8.7Hz, 2H), 1.66 (s, 6H). MS (ESI) m / z: 394.2 [MH] ^- .

Step 11: Synthesis of 4-benzyloxy-1,2-dihydrofuro [3,2-f] quinolin-9-ol

At room temperature, 5-((7-benzyloxy-2,3-dihydrobenzofuran-5-ylamino) methylene) -2,2-dimethyl-1 was sequentially added to the microwave reaction flask. 3-Dioxane-4,6-dione (1.5 g, 3.79 mmol) and diphenyl ether (18 ml) were stirred in a 70 ° C water bath for 15 minutes, and then transferred to a microwave at 220 ° C for 0.5 h to terminate. Cool to room temperature, add PE to beat for 1 h, and suction filter to obtain 1.02 g of yellow solid with a yield of 92%. Without further purification, it was directly used in the next reaction. ¹ H NMR (400MHz, DMSO-d ₆ ) δ7.67 (t, J = 4.0Hz, 1H), 7.49–7.33 (m, 5H), 6.92 (s, 1H), 5.83 (d, J = 7.2Hz, 1H), 5.19 (s, 2H), 4.58 (t, J = 9.1Hz, 2H), 3.66 (t, J = 9.0Hz, 2H). MS (ESI) m / z: 294.2 [M + H] ⁺ .

Step 12: Synthesis of 4-benzyloxy-9-chloro-1,2-dihydrofuro [3,2-f] quinoline

At room temperature, 4-benzyloxy-1,2-dihydrofuro [3,2-f] quinolin-9-ol (6.17 g, 21.04 mmol), and phosphorus oxychloride (60 ml) were sequentially added to the three-necked flask. ), N _{2 was} replaced three times, and the reaction was terminated at 107 ° C. for 0.5 h. It was concentrated to obtain a black crude product, which was purified by column chromatography (MeOH / DCM system) to obtain 3.62 g of a yellow solid with a yield of 55%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.51 (d, J = 4.8 Hz, 1 H), 7.51–7.45 (m, 4H), 7.44–7.33 (m, 3H), 5.32 (s, 2H), 4.76 (t, J = 9.2 Hz, 2H), 3.88 (t, J = 9.2 Hz, 2H). MS (ESI) m / z: 312.2 [M + H] ⁺ .

Step 13: 4-benzyloxy-9- (4-fluoro-2methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2-f] quinoline Synthesis

At room temperature, 4-benzyloxy-9-chloro-1,2-dihydrofuro [3,2-f] quinoline (3.62 g, 11.61 mmol), 2-methyl- 4-Fluoro-5-hydroxy-1-hydroindole (3.84g, 23.25mmol), DMAP (2.41g, 19.73mmol), 2,6-dimethylpyridine (72ml), N ₂ substitution three times, microwave 200 ° C The reaction was terminated in 2.5 h. After concentration, it was extracted with EA, washed with saturated brine twice, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (THF / DCM system) to obtain 1.29 g of a yellow solid with a yield of 25%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.35 (d, J = 5.2 Hz, 1H), 7.52 (d, J = 7.2 Hz, 2H), 7.45–7.34 (m, 4H), 7.20 (d, J = 8.6Hz, 1H), 6.98 (t, J = 8.1Hz, 1H), 6.31 (d, J = 5.1Hz, 1H), 6.27 (s, 1H), 5.31 (s, 2H), 4.75 (t, J = 9.1Hz, 2H), 3.81 (t, J = 9.1Hz, 2H), 2.41 (s, 3H). MS (ESI) m / z: 441.2 [M + H] ⁺ .

Step 14: Synthesis of 9- (4-fluoro-2methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2-f] quinolin-4-ol

At room temperature, 4-benzyloxy-9- (4-fluoro-2methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2 -f] quinoline (0.62 g, 1.41 mmol), N ₂ substitution three times, TFA (12 ml), H ₂ O (3 ml) were added, and the reaction was terminated at 85 ° C. for 3 h. It was extracted with a mixed solution of EA and THF, washed twice with saturated brine, and the aqueous phase was back-extracted once with EA. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (EA / MeOH system) to obtain a yellow solid. 0.31 g, yield 50%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.28 (d, J = 5.2 Hz, 1 H), 7.19 (d, J = 8.6 Hz, 1 H), 7.17 (s, 1 H), 6.97 (t, J = 8.1Hz, 1H), 6.26 (s, 1H), 6.22 (d, J = 5.1Hz, 1H), 4.73 (t, J = 9.1Hz, 2H), 3.79 (t, J = 9.1Hz, 2H), 2.41 (s, 3H). MS (ESI) m / z: 351.2 [M + H] ⁺ .

Step 15: Synthesis of (1- (tert-butoxycarbonylamino) cyclopropyl) methyl mesylate

Add (1-hydroxymethylcyclopropyl) -tert-butoxycarbonylamino (1.00 g, 5.34 mmol), DCM (10 ml), DIEA (1.38 g, 10.68 mmol) to a single-necked flask under an ice bath, and stir for 15 min MsCl (0.673 g, 5.85 mmol) was added dropwise, and the reaction was terminated at room temperature for 0.5 h after the dropwise addition. Extracted with EA, washed twice with saturated sodium bicarbonate solution, dried the organic phase over anhydrous sodium sulfate, filtered, and concentrated to give 1.29 g of a pale yellow solid with a yield of 91%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 4.18 (s, 2H), 3.12 (s, 3H), 1.37 (s, 9H), 0.79 (t, J = 4.0Hz, 2H), 0.73 (t, J = 4.0Hz, 2H).

Step 16: 1-((9- (4-fluoro-2methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2-f] quinoline-4 -Yloxy) methyl) cyclopropylcarbamic acid tert-butyl ester synthesis

At room temperature, 9- (4-fluoro-2-methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2-f] quinoline was added to a three-necked flask. 4-alcohol (0.10 g, 0.29 mmol), (1- (tert-butoxycarbonylamino) cyclopropyl) methyl methanesulfonate (0.30 g, 1.13 mmol), cesium carbonate (0.46 g, 1.41 mmol), DMAC (6ml), N ₂ three times replacement, 100 ℃ 1h reaction was terminated. Extracted with EA, washed twice with purified water, dried the organic phase over anhydrous sodium sulfate, filtered, concentrated, and then purified by forward column chromatography (MeOH / DCM), followed by reverse phase column purification (0.05% aqueous phosphoric acid solution / acetonitrile system) 0.07 g of a yellow solid was obtained with a yield of 50%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 7.35 (s, 1H), 7.27 (s, 1H), 7.20 (d, J = 8.6Hz, 1H), 6.98 (t, J = 8.1Hz, 1H) , 6.30 (d, J = 5.0Hz, 1H), 6.27 (s, 1H), 4.75 (t, J = 9.1Hz, 2H), 4.17 (s, 2H), 3.81 (t, J = 9.2Hz, 2H) , 2.41 (s, 3H), 1.36 (s, 9H), 0.85 (t, J = 5.6Hz, 2H), 0.76 (t, J = 5.6Hz, 2H). MS (ESI) m / z: 520.2 [M + H] ⁺ .

Step 17: 1-((9- (4-fluoro-2-methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2-f] quinoline- Synthesis of 4-yloxy) methyl) -cyclopropane-1-aminohydrochloride

At room temperature, 1-((9- (4-fluoro-2methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2- f] Quinolin-4-yloxy) methyl) cyclopropylcarbamic acid tert-butyl ester (0.04g, 0.08mmol), THF (2ml), N ₂ replacement three times, tetrahydrofuran hydrochloride solution was added dropwise under ice bath, dropwise After completion, the reaction was terminated at 40 ° C for 3h. After moving to an ice bath and stirring for 0.5 h, 0.03 g of yellow solid was obtained by suction filtration with a yield of 70%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.72 (d, J = 6.3 Hz, 1 H), 7.72 (s, 1 H), 7.29 (d, J = 8.4 Hz, 1 H), 7.11 (t, J = 7.9Hz, 1H), 6.79 (d, J = 6.0Hz, 1H), 6.32 (s, 1H), 4.90 (t, J = 9.1Hz, 2H), 4.45 (s, 2H), 3.93 (t, J = 9.0Hz, 2H), 2.43 (s, 3H), 1.19 (t, J = 3.0Hz, 2H), 1.07 (t, J = 3.0Hz, 2H). MS (ESI) m / z: 418.2 [MH] ^- .

Example 2

9- (4-fluoro-2-methyl-1-hydroindole-5-yloxy) -4- (2- (pyrrolidin-1-yl) -hydroxyethyl) -1,2-dihydro Furo [3,2-f] quinoline

Step 1: Referring to steps 1-14 of Example 1, 9- (4-fluoro-2methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2- f] quinolin-4-ol.

Step 2: Synthesis of 2- (pyrrolidin-1-ylethyl) methanesulfonate

At room temperature, N- (2-hydroxyethyl) -pyrrolidine (1.00 g, 8.68 mmol), NaHCO ₃ (1.75 g, 20.83 mmol), DCM (15 ml), MsCl (0.99 g, 8.64) mmol), the reaction was terminated at room temperature for 3h. Anhydrous sodium sulfate was added to the reaction solution, stirred to dry, filtered, and concentrated to obtain a yellow solid, which was slurried with DCM and filtered with suction to obtain a white solid and a filtrate. The filtrate was concentrated and purified by column chromatography (ammonia methanol solution / DCM system) 0.22 g of pale yellow solid with a yield of 13%.

Step 3: 9- (4-fluoro-2-methyl-1-hydroindole-5-yloxy) -4- (2- (pyrrolidin-1-yl) -hydroxyethyl) -1,2 -Dihydrofuro [3,2-f] quinoline Synthesis

At room temperature, 9- (4-fluoro-2methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2-f] quinoline was sequentially added to the three-necked flask. 4-alcohol (0.05g, 0.14mmol), 2- (pyrrolidin-1-ylethyl) methanesulfonate (0.06g, 0.31mmol), Cs ₂ CO ₃ (0.12g, 0.37mmol), DMAC ( 3 ml), N _{2 was} replaced three times, and the reaction was terminated at 100 ° C. for 1 h. Extracted with EA, washed twice with purified water, dried the organic phase over anhydrous sodium sulfate, filtered, concentrated, purified by forward column chromatography (ammonia methanol solution / MeOH), and then reversed-phase column purification (0.1% formic acid aqueous solution / acetonitrile system) ), And finally purified by column chromatography (ammonia methanol solution / MeOH) to obtain 0.01 g of off-white solid with a yield of 15%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.36 (d, J = 5.2 Hz, 1 H), 7.33 (s, 1 H), 7.20 (d, J = 8.6 Hz, 1 H), 6.98 (t, J = 8.0Hz, 1H), 6.30 (d, J = 5.1Hz, 1H), 6.27 (s, 1H), 4.75 (t, J = 9.0Hz, 2H), 4.28 (t, J = 4.9Hz, 2H), 3.81 (t, J = 9.1 Hz, 2H), 2.92 (t, J = 4.0 Hz, 2H), 2.67–2.57 (m, 4H), 2.41 (s, 3H), 1.75–1.69 (m, 4H). MS (ESI) m / z: 448.2 [M + H] ⁺ .

Example 3

2- (9- (4-fluoro-2-methyl-1H-indol-5-yl) oxy) -1,2-dihydrofuran [3,2-f] quinolin-4-yl) ethyl (Alkoxy) -1-ol

At room temperature, add 9- (4-fluoro-2methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2-f] quinoline- 4-Alcohol (0.20 g, 0.57 mmol), 2-bromoethanol (0.29 g, 2.28 mmol), Cs ₂ CO ₃ (0.93 g, 2.85 mmol) and DMAC (5 mL), protected by nitrogen, and terminated at 60 ° C. for 2.5 h. Extracted by EA, washed twice with saturated brine, and dried the organic phase over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (MeOH / DCM system) to obtain 0.06 g of a yellow solid product with a yield of 28%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.35 (d, J = 5.1 Hz, 1 H), 7.31 (s, 1 H), 7.20 (d, J = 8.5 Hz, 1 H), 6.98 (t, J = 8.0Hz, 1H), 6.30 (d, J = 4.9Hz, 1H), 6.27 (s, 1H), 4.74 (t, J = 9.0Hz, 2H), 4.18 (t, J = 4.2Hz, 2H), 3.81 (t, J = 7.0 Hz, 4H), 2.41 (s, 3H). MS (ESI) m / z: 395.0 [M + H] ⁺ .

Example 4

(S) -1-((9-((4-fluoro-2-methyl-1H-indol-5-yl) oxy) -1,2-dihydrofuran [3,2-f] quinoline -4-yl) oxy) propane-2-ol

At room temperature, 9- (4-fluoro-2-methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2-f] quinoline was added to a three-necked flask. 4-alcohol (0.20 g, 0.57 mmol), Cs ₂ CO ₃ (0.93 g, 2.85 mmol), (S) -1-chloro-2-propanol (0.22 g, 2.28 mmol) and DMAC (3 mL), nitrogen Protected and terminated at 80 ° C for 2h. EA was added for extraction, and saturated brine was washed twice with water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (MeOH / DCM system) to obtain 0.98 g of a yellow solid product with a yield of 70%. MS (ESI) m / z: 409.2 [M + H] ⁺ .

Example 5

(R) -1-((9-((4-fluoro-2-methyl-1H-indol-5-yl) oxy) -1,2-dihydrofuran [3,2-f] quinoline -4-yl) oxy) propane-2-ol

At room temperature, add 9-((4-fluoro-2methyl-1-hydroindol-5-yl) oxy) -1,2-dihydrofuro [3,2-f] quine to a three-necked flask. Porphyrin-4-ol (0.20g, 0.57mmol), Cs ₂ CO ₃ (0.93g, 2.85mmol), (R) -1-chloro-2-propanol (0.22g, 2.28mmol) and DMAC (3mL), Under nitrogen protection, the reaction was terminated at 80 ° C for 2h. EA was added for extraction, the saturated brine was washed twice with water, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (MeOH / DCM system) to obtain 0.13 g of a yellow solid product with a yield of 55%. MS (ESI) m / z: 409.2 [M + H] ⁺ .

Example 6

(R) -3-((9-((4-fluoro-2-methyl-1H-indol-5-yl) oxy) -1,2-dihydrofuran [3,2-f] quinoline -4-yl) oxy) propane-1,2-diol

At room temperature, 9- (4-fluoro-2-methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2-f] quinoline was added to a three-necked flask. 4-alcohol (0.20 g, 0.57 mmol), Cs ₂ CO ₃ (0.93 g, 2.85 mmol), (R) -3-chloro-1,2-propanediol (0.25 g, 2.28 mmol) and DMF (3 mL), Under nitrogen protection, the reaction was terminated at 80 ° C for 4h. EA was added for extraction, the saturated brine was washed twice with water, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (MeOH / DCM system) to obtain 0.09 g of a yellow solid product with a yield of 37%. MS (ESI) m / z: 425.1 [M + H] ⁺ .

Example 7

(S) -3-((9-((4-fluoro-2-methyl-1H-indol-5-yl) oxy) -1,2-dihydrofuran [3,2-f] quinoline -4-yl) oxy) propane-1,2-diol

At room temperature, 9- (4-fluoro-2-methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2-f] quinoline was added to a three-necked flask. 4-alcohol (0.20 g, 0.57 mmol), Cs ₂ CO ₃ (0.93 g, 2.85 mmol), (S) -3-chloro-1,2-propanediol (0.25 g, 2.28 mmol) and DMF (3 mL), Under nitrogen protection, the reaction was terminated at 80 ° C for 4h. EA was added for extraction, and saturated brine was washed twice with water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (MeOH / DCM system) to obtain 0.10 g of a yellow solid product with a yield of 41%. MS (ESI) m / z: 425.1 [M + H] ⁺ .

Example 8

2- (2-((3S, 4R) -3,4-dimethoxypyrrolidin-1-yl) ethoxy) -9-((4-fluoro-2-methyl-1H-indole- 5-yl) oxy) -1,2-dihydrofuran [3,2-f] quinoline

Step 1: Synthesis of tert-butyl (3R, 4S) -3,4-dimethoxypyrrolidine-1-carboxylic acid ester

In an ice bath, add tert-butyl (3R, 4S) -3,4-dihydroxypyrrolidine-1-carboxylic acid ester (1.00 g, 4.92 mmol), NaH (0.30 g, 12.30 mmol) and DMF to a three-necked flask. (10 mL), protected by nitrogen, stirred at 0 ° C. for 15 min, and then added iodomethane (1.54 g, 10.83 mmol) dropwise. After the dropwise addition, the reaction was terminated at 0 ° C. for 3 h. EA was added for extraction, and saturated brine was washed twice with water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. Purified by column chromatography (PE / EA system) to obtain 0.91 g of a pale yellow oil with a yield of 80%.

Step 2: Synthesis of (3R, 4S) -3,4-dimethoxypyrrolidine hydrochloride

At room temperature, add tert-butyl (3R, 4S) -3,4-dimethoxypyrrolidine-1-carboxylic acid ester (0.91g, 3.93mmol) and DCM (5mL) to a single-necked flask, and drop in an ice bath. A solution of hydrochloric acid in ethyl acetate (1 mL) was added, and the reaction was terminated at 20 ° C. for 1 h after the dropwise addition. Concentration gave 0.65 g of a yellow solid with a yield of 98%.

Step 3: 4- (2-Bromoethoxy) -9-((4-fluoro-2-methyl-1H-indol-5-yl) oxy) -1,2-dihydrofuran [3, Synthesis of 2-f] quinoline

At room temperature, 9- (4-fluoro-2-methyl-1-hydroindole-5-yloxy) -1,2-dihydrofuro [3,2-f] quinoline was added to a three-necked flask. 4-alcohol (1.00g, 2.85mmol), K ₂ CO ₃ (1.18g, 8.56mmol), 1,2-dibromoethane (1.61g, 8.56mmol) and DMAC (5mL), nitrogen protected, 90 ° C The reaction was terminated 6h. EA was added for extraction, and saturated brine was washed twice with water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (MeOH / DCM system) to obtain 0.98 g of a yellow solid with a yield of 70%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.41 (d, J = 5.3 Hz, 1H), 7.35 (s, 1H), 7.21 (d, J = 8.6 Hz, 1H), 7.00 (t, J = 8.1Hz, 1H), 6.38 (d, J = 5.2Hz, 1H), 6.27 (s, 1H), 4.78 (t, J = 9.2Hz, 2H), 4.54 (t, J = 5.3Hz, 2H), 3.89 (t, J = 5.3 Hz, 2H), 3.83 (t, J = 9.1 Hz, 2H), 2.41 (s, 3H). MS (ESI) m / z: 456.8, 458.8 [M + H] ⁺ .

Step 4: 4- (2-((3S, 4R) -3,4-dimethoxypyrrolidin-1-yl) ethoxy) -9-((4-fluoro-2-methyl-1H- Synthesis of indole-5-yl) oxy) -1,2-dihydrofuran [3,2-f] quinoline

Add 4- (2-bromoethoxy) -9-((4-fluoro-2-methyl-1H-indol-5-yl) oxy) -1,2-di Hydrofuran [3,2-f] quinoline (0.20 g, 0.44 mmol), (3R, 4S) -3,4-dimethoxypyrrolidine hydrochloride (0.11 g, 0.66 mmol), Cs ₂ CO ₃ (0.43 g, 1.31 mmol) and DMF (8 mL), protected by nitrogen, terminated at 80 ° C. for 2 h. EA was added for extraction, and the saturated brine was washed twice with water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and purified by reversed-phase column (0.05% phosphoric acid aqueous solution / acetonitrile system) to obtain 0.14 g of a white solid product with a yield of 64%. MS (ESI) m / z: 508.1 [M + H] ⁺ .

Example 9

4- (2-((3R, 4R) -3,4-dimethoxypyrrolidin-1-yl) ethoxy) -9-((4-fluoro-2-methyl-1H-indole- 5-yl) oxy) -1,2-dihydrofuran [3,2-f] quinoline

Step 1: Synthesis of tert-butyl (3S, 4S) -3,4-dimethoxypyrrolidine-1-carboxylic acid ester

Add tert-butyl (3S, 4S) -3,4-dihydroxypyrrolidine-1-carboxylic acid ester (1.00g, 4.92mmol), NaH (0.30g, 12.30mmol) and DMF to a three-necked flask under ice bath (10 mL), protected by nitrogen, stirred at 0 ° C. for 15 min, and then added iodomethane (1.54 g, 10.83 mmol) dropwise. After the dropwise addition, the reaction was terminated at 0 ° C. for 3 h. EA was added for extraction, and the saturated brine was washed twice with water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and purified by column chromatography (PE / EA system) to obtain 0.80 g of a pale yellow oil with a yield of 70%.

Step 2: Synthesis of (3S, 4S) -3,4-dimethoxypyrrolidine hydrochloride

At room temperature, add tert-butyl (3S, 4S) -3,4-dimethoxypyrrolidine-1-carboxylic acid ester (0.80 g, 3.93 mmol) and DCM (5 mL) to a single-necked flask, and drop in an ice bath. A solution of hydrochloric acid in ethyl acetate (1 mL) was added, and the reaction was terminated at 20 ° C. for 1 h after the dropwise addition. Concentration gave 0.55 g of a yellow solid with a yield of 95%.

Step 3: 4- (2-((3R, 4R) -3,4-dimethoxypyrrolidin-1-yl) ethoxy) -9-((4-fluoro-2-methyl-1H- Synthesis of indole-5-yl) oxy) -1,2-dihydrofuran [3,2-f] quinoline

Add 4- (2-bromoethoxy) -9-((4-fluoro-2-methyl-1H-indol-5-yl) oxy) -1,2-di Hydrofuran [3,2-f] quinoline (0.40 g, 0.88 mmol), (3S, 4S) -3,4-dimethoxypyrrolidine hydrochloride (0.22 g, 1.31 mmol), Cs ₂ CO ₃ (0.86g, 2.62mmol) and DMF (20mL), protected by nitrogen, the reaction was terminated at 80 ° C for 2h. EA was added for extraction, and saturated brine was washed twice with water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and purified by reversed-phase column (0.05% phosphoric acid aqueous solution / acetonitrile system) to obtain 0.26 g of off-white solid product with a yield of 60%. MS (ESI) m / z: 508.1 [M + H] ⁺ .

Example 10

VEGFR kinase activity test

Inhibition This test uses γ- ³³ p-ATP isotope test compound on kinase assays VEGFR / VEGFR2 / VEGFR3, and to obtain the compound inhibiting the activity of half inhibitory concentration IC _50. The positive control drug Anlotinib was purchased from Suzhou Hengning Pharmaceutical Technology Co., Ltd., batch number 2016120201, and Lenvatinib was purchased from Hubei Weidley Chemical Technology Co., Ltd., batch number HBW171008.

Basal reaction buffer

20mM Hepes (pH 7.5), 10mM MgCl ₂ , 1mM EGTA, 0.02% Brij35, 0.02mg / ml BSA, 0.1mM Na3VO4, 2mM DTT, 1% DMSO.

2. Compound formulation

The compound was dissolved to a specific concentration with 100% DMSO, and then it was gradient diluted to a different concentration of the test sample (DMSO solution) using an automatic sampling device.

3. Reaction steps

3.1 Dilute the reaction substrate with the basic reaction buffer;

3.2 Add the kinase to the substrate solution and mix gently;

3.3 Use an automatic sample loading system to add 100% DMSO-diluted compounds of different concentrations to the kinase solution and incubate for 20 min at room temperature;

3.4 Add ³³ P-ATP (10 μM, 10 μCi / μl) at room temperature to start the kinase reaction, and react for 2 h.

4. Detection

The reaction solution was subjected to ion exchange filtration system to remove unreacted ATP and ADP plasma generated by the reaction, and then measured the ³³ P isotope radiation in the substrate.

5. Data processing

The inhibitory effect of different concentrations of compounds on kinase activity was calculated based on the amount of kinase added to the inhibitor system at different concentrations, and the IC ₅₀ was inhibited by graphpad prism fitting.

The biochemical activity of the compounds of the present invention was determined through the above tests. The measured IC ₅₀ values are shown in Table 1:

Table 1 VEGFR kinase activity test results

Note: "-" in the table means not tested.

Conclusion: Compared with the positive control drug Anlotinib, the compound of the present invention has better kinase inhibitory activity, especially VEGFR2 inhibitory activity.

Example 11

VEGF-induced HUVEC cell proliferation inhibition test

This test method by MTT test compound effect of VEGF-induced HUVEC cell activity and derived compounds inhibit VEGF-induced proliferation of HUVEC cells half maximal inhibitory concentration IC ₅₀ activity.

1. HUVEC cell lines were cultured under the conditions of ECM + 5% FBS + 1% ECGS (endothelial cell growth factor additive) + 1% P / S (penicillin streptomycin mixed solution). The day before the experiment, the cells were cultured with ECM + 5% FBS. A 96-well cell culture plate was inoculated with 100 μL of HUVEC cell suspension in logarithmic growth phase, with a density of 5 × 10 ⁴ / ml. The culture plate was cultured in an incubator for 24 hours to make the cells adhere (37 ° C, 5% CO ₂ ).

2. Each compound has been dissolved in DMSO to prepare a 10 mM stock solution, which is diluted 400 times the target concentration with DMSO and diluted to 2 times the target concentration with serum-free medium to maintain the DMSO concentration in the drug solution at 0.5. %. In a 96-well plate inoculated with cells, different concentrations of drug solution and 10 ng / mL of VEGF165, 100 μL / well were sequentially added. Three replicates were set for each concentration, and blank control, normal control, and VEGF-induced groups were set up, and the culture was continued at 37 ° C and 5% CO ₂ for 72 hours.

3. Terminate the culture, add 20 μL of MTT solution (5mg / ml) to each well, continue to incubate at 37 ° C, 5% CO ₂ for 4 h, discard the medium, add DMSO 150 μL / well, shake at room temperature for 10 min, and 570 nM, 620 nM OD values (OD _570- OD ₆₂₀ ) were measured at two wavelengths, and IC ₅₀ values were calculated by Graphpad Prism 6.0 data processing.

The biochemical activity of the compounds of the present invention was determined through the above tests. The measured IC ₅₀ values are shown in Table 2:

Table 2 VEGF-induced HUVEC cell proliferation inhibition test

Conclusion: Compared with the positive control, the compound 2 of the present invention has better activity of inhibiting the proliferation of HUVEC cells induced by VEGF.

Example 12

Liver microsome test

The total volume of the incubation system was 250 μL. A 50 mmol / L PBS buffer (pH = 7.4) was used to prepare a human liver microsome incubation solution with a protein concentration of 0.5 mg / mL. Before incubation, 2.5 μL of 100 μmol / L test compound and The above incubation solution was mixed with 197.5 μL, and pre-incubated in a 37 ° C water bath for 5 min. Then, 50 μL of a reduced coenzyme Ⅱ solution (5 mmol / L) which was also pre-incubated for 5 min was added to start the reaction. g / L, final concentration of test compound is 1 μmol / L), incubate with shaking in a 37 ° C water bath, and take them out at 0, 5, 15, 30, and 60 min respectively, and immediately add the internal standard as Terfenadine (positive ion internal standard, 25ng / mL) and Tolbutamide (anion internal standard, 50ng / mL) positive and negative internal standard mixed methanol solution 600 μL to stop the reaction. The incubation solution was shaken for 2 minutes, centrifuged (4 ° C, 16000 r / min) for 10 minutes, and the supernatant was taken for LC-MS / MS detection to quantitatively analyze the remaining amount of the parent drug. (DMSO <0.1%).

The concentration of the compound at 0 min incubation was taken as 100%, and the concentrations at other incubation time points were converted to the percentage remaining amount. The natural logarithm of the percentage remaining amount at each time point was linearly regression to the incubation time, and the slope k was calculated. According to the formula , T _1/2 = -0.693 / k calculated in vitro half-life. Clearance in liver microsomes (CLint (μL / min / mg protein) = Ln (2) * 1000 / T _1/2 (min) / Protein Conc (mg / ml)).

The liver microsomal test data is shown in Table 3:

Table 3 Liver microsomal test results

Conclusion: Compared with the positive control drug, the compound of the present invention has considerable human liver microsomal metabolic stability, and therefore has good drug-making properties.

Claims (10)

A compound represented by formula (I), a stereoisomer thereof, a pharmaceutically acceptable salt or ester, or a solvate:

R ^{1 is} selected from C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, nitro, -NR ⁸ R ⁹ ,- NR ⁸ COR ⁷ , -COR ⁷ , -SO ₂ R ⁷ , -SOR ⁷ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₈ alkoxy or 4-10 membered heterocyclic The C ₃ -C ₆ cycloalkyl or 4- to 10-membered heterocyclic group may be substituted by an amino group, a hydroxyl group, a-(CH ₂ ) _n CN, a carboxyl group, or a C ₁ -C ₄ alkyl group. Or C ₁ -C ₄ alkoxy. n is selected from 0, 1, 2 or 3; R ^{2 is} selected from H, deuterium or halogen; X is selected from N or CH; Y is selected from O or CR ³ R ⁴ ; R ³ and R ⁴ are each independently selected from H, deuterium, halogen, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or C ₁ -C ₈ alkoxy; R ^{5 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl; R ^{6 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl; R ^{7 is} selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, -NR ⁸ R ⁹ or C ₃ -C ₆ heterocyclyl. The C ₃ -C ₆ heterocyclyl may be Further substituted with hydroxy, carboxy or C ₁ -C ₄ alkyl; R ⁸ and R ⁹ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl. The compound according to claim 1, wherein: R ¹ is C ₁ -C ₄ alkyl, optionally further substituted with one or more substituents selected from deuterium, hydroxyl, methyl, ethyl, cyclopropyl, or a 4-6 membered heterocyclic group, said The cyclopropyl or 4- to 6-membered heterocyclic group may be further substituted with an amino group, a hydroxyl group, a carboxyl group, a C _{1 to} C ₄ alkyl group, or a C _{1 to} C ₄ alkoxy group. Is selected from pyrrolidinyl, morpholinyl, piperazinyl or piperidinyl. R ^{2 is} selected from H, deuterium or halogen; X is CH; Y is CH ₂ ; R ³ and R ⁴ are each independently selected from H, deuterium, halogen, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or C ₁ -C ₈ alkoxy; R ^{5 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl; R ^{6 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl. A compound according to general formula (I), a stereoisomer thereof, a pharmaceutically acceptable salt or ester or a solvate according to claim 1, which is a compound according to general formula (II), a stereoisomer thereof, Pharmaceutically acceptable salts or esters or solvates:

R ¹ is C ₁ -C ₈ alkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, -COR ⁷ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl Or a 4-6 membered heterocyclyl substituent, the C ₃ -C ₆ cycloalkyl or 4-6 membered heterocyclyl may be further substituted with amino, hydroxyl,-(CH ₂ ) _n CN, carboxyl, C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy; n is selected from 0, 1, 2 or 3; R ^{7 is} selected from the group consisting of H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or C ₃ -C ₆ heterocyclyl. The C ₃ -C ₆ heterocyclyl may be further substituted by hydroxy, carboxy or C ₁ -C ₄ alkyl. The compound according to claim 3, wherein: R ¹ is C ₁ -C ₄ alkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, C ₁ -C ₂ alkyl, C ₃ -C ₆ cycloalkyl, or 4-6 membered heterocyclic group substituents, the C ₃ -C ₆ cycloalkyl or 4-6 membered heterocyclic group may be further substituted by amino, _{hydroxy, - (CH 2) n CN} , carboxy, C ₁ -C Substituted with ₄ alkyl or C ₁ -C ₄ alkoxy; n is selected from 0, 1, or 2. The compound according to claim 4, characterized in that: R ¹ is C ₁ -C ₄ alkyl, optionally further substituted with one or more substituents selected from deuterium, hydroxyl, methyl, ethyl, cyclopropyl, or a 4-6 membered heterocyclic group, said The cyclopropyl or 4- to 6-membered heterocyclic group may be further substituted with an amino group, a hydroxyl group, a carboxyl group, a C _{1 to} C ₄ alkyl group, or a C _{1 to} C ₄ alkoxy group. Is selected from pyrrolidinyl, morpholinyl, piperazinyl or piperidinyl. A compound, a stereoisomer thereof, a pharmaceutically acceptable salt or ester, or a solvent, characterized in that the compound is selected from:

A pharmaceutical composition comprising the compound according to any one of claims 1 to 6, a stereoisomer thereof, a pharmaceutically acceptable salt or ester or a solvate, and a pharmaceutically acceptable carrier. The pharmaceutical composition according to claim 7, wherein the pharmaceutical composition is a capsule, powder, tablet, granule, pill, injection, syrup, oral solution, inhalant, ointment, suppository or Patch. Use of the compound according to any one of claims 1 to 6 or the pharmaceutical composition according to any one of claims 7 to 8 in the manufacture of a therapeutic medicament for preventing or treating an angiogenesis-mediated disease. Use of the compound according to any one of claims 1 to 6 or the pharmaceutical composition according to any one of claims 7 to 8 in the preparation of a medicament for preventing or treating malignant tumors.

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