(S)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide derivatives and their applications

Abstract

The present invention discloses an (S)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide derivative and its application. The (S)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide derivative includes a compound represented by general formula (I), a stereoisomer of general formula (I) or a salt thereof: wherein the R group is any one of a heteroaryl group, a heterocyclic group, an oxo heterocyclic group, a thio heterocyclic group, a substituted aryl group, a substituted heteroaryl group, a substituted heterocyclic group, and a substituted amino group.

Description

Technical Field

[0001] The present invention belongs to the technical field of anti-cancer drugs, and relates to a (S)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide derivative and its application. Background Art

[0002] Phosphoinositide 3-kinase (PI3K) belongs to the lipid kinase family and can catalyze the phosphorylation of the 3-hydroxy group of phosphatidylinositol to generate phosphatidylinositol-3,4,5-trisphosphate (PtdIns(3,4,5)P3, PIP3). As a second messenger, PIP3 can activate protein kinase B (AKT) or other cellular messengers such as mammalian target of rapamycin (mTOR). The PI3K / AKT / mTOR signaling pathway is involved in regulating processes such as cell growth, survival, proliferation, apoptosis, and migration. Therefore, the dysregulation or mutation of the PI3K / AKT / mTOR signaling pathway can lead to cancer. PI3K kinases can be divided into three classes, Class I, II, and III, according to their structural characteristics and substrate specificities. Currently, the research on Class I PI3K is the most in-depth, and Class I PI3K can be divided into IA and IB according to different regulatory methods. Class IA PI3K contains three subtypes, PI3Kα, PI3Kβ, and PI3Kδ, and is a heterodimer composed of catalytic subunits p110α, p110β, p110δ and regulatory subunits p85α, p85β, p55α, p55δ, and p50α. Class IB PI3K is a heterodimer composed of catalytic subunit p110γ and one of the regulatory subunits p101, p87, or p84. The mutation of PI3Kα is closely related to the occurrence and development of cancer. Amplification and somatic missense mutations of the PIK3CA gene encoding PI3Kα have been found in different types of cancers such as colon cancer, gastric cancer, cervical cancer, ovarian cancer, prostate cancer, lung cancer, and breast cancer. By comparing the distribution of PIK3CA mutation forms in tumors, lung cancer, ovarian cancer, head and neck cancer, esophageal cancer, and prostate cancer are mainly characterized by PIK3CA amplification, while breast cancer, uterine cancer, colorectal cancer, and bladder cancer are mainly characterized by PIK3CA mutation. These mutations or amplifications will lead to the abnormal activation of PI3Kα and excessive cell proliferation, thus resulting in tumorigenesis. PI3Kα is an important target for anti-cancer drug research and development, and PI3Kα selective inhibitors can be used to treat a variety of multiple tumors with PI3Kα activation mutations or amplifications, and have great clinical application value. 2-Aminothiazole-based PI3Kα selective inhibitors are used to treat hormone receptor-positive (HR + ) and human epidermal growth factor receptor 2-negative (HER2 -) Advanced or metastatic breast cancer with PIK3CA mutations, but with low selectivity for the PI3Kα kinase during treatment, thus accompanied by side effects such as hyperglycemia, diarrhea, decreased appetite, rash, etc. Therefore, there is an urgent need to research and develop new-structured PI3Kα selective inhibitors with high activity and high selectivity. Summary of the Invention

[0003] In view of the problems existing in the prior art, the present invention provides a (S)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide derivative and its application, thereby solving the technical problems of low inhibitory activity and low selectivity of existing drugs for PI3Kα.

[0004] The present invention is achieved through the following technical solutions:

[0005] A (S)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide derivative, including a compound represented by the general formula (I), a stereoisomer of the general formula (I), or a salt thereof:

[0006]

[0007] Among them, the R group is any one of a heteroaryl group, a heterocyclic group, an oxoheterocyclic group, a thioheterocyclic group, a substituted aryl group, a substituted heteroaryl group, a substituted heterocyclic group, and a substituted amino group.

[0008] Preferably, the R group is:

[0009] And substituted Any one of them.

[0010] Preferably, the R group is:

[0011]

[0012] Any one of them. Preferably, the R group is:

[0013]

[0014]

[0015] Any one of them.

[0016] Preferably, the R group is:

[0017]

[0018] Any one of the following.

[0019] Preferably, the R group is:

[0020]

[0021] any one of

[0022] A pharmaceutical composition comprising a compound represented by the general formula (I) as described in any one of the above, a stereoisomer of the general formula (I) or a salt thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients; the pharmaceutical composition is made into tablets, capsules, soft capsules or injections by adding excipients to the compound, its stereoisomer or its salt as shown above; the excipients include one or several of additives, stabilizers, solubilizers, lubricants and disintegrants.

[0023] Use of the compound represented by the general formula (I) as described above, a stereoisomer of the general formula (I) or a salt thereof, or a pharmaceutical composition in the preparation of a PI3Kα inhibitor drug.

[0024] Use of the compound represented by the general formula (I) as described above, a stereoisomer of the general formula (I) or a salt thereof, or a pharmaceutical composition in the preparation of a cancer drug; the cancer includes any one of breast cancer, pancreatic cancer, non-small cell lung cancer, thyroid cancer, seminoma, melanoma, bladder cancer, liver cancer, gastric cancer, kidney cancer, ovarian cancer, uterine cancer, prostate cancer, glioblastoma, myelodysplastic syndrome, acute myeloid leukemia and colorectal cancer.

[0025] A method for preparing the compound represented by the general formula (I) as described above, a stereoisomer of the general formula (I) or a salt thereof, characterized in that

[0026] when the R is any one of heteroaryl, substituted aryl, substituted heteroaryl, oxoheterocyclic group or thioheterocyclic group, it comprises the following steps:

[0027] S1: React 2,4-dibromobenzoic acid with resorcinol to obtain 3-hydroxy-9-bromo-6H-dibenzo[b,d]pyran-6-one;

[0028] S2: Reduce 3-hydroxy-9-bromo-6H-dibenzo[b,d]pyran-6-one by 1,1,3,3-tetramethyldisiloxane to obtain 9-bromo-6H-dibenzo[b,d]pyran-3-ol;

[0029] S3: React 9-bromo-6H-dibenzo[b,d]pyran-3-ol with (R)-methyl 2-hydroxypropionate to prepare methyl (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate;

[0030] S4: React methyl (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate with an ammonia methanol solution to obtain (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide;

[0031] S5: React (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide with bis(pinacolato)diboron to obtain (S)-2-[(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide;

[0032] S6: React (S)-2-[(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide with any one of a halogenated heteroaryl, a halogenated substituted aryl, a halogenated substituted heteroaryl, a halogenated oxoheterocyclic group, or a halogenated thioheterocyclic group to obtain a compound of general formula (I), a stereoisomer thereof, or a salt thereof, wherein R is any one of a heteroaryl, a substituted aryl, a substituted heteroaryl, an oxoheterocyclic group, or a thioheterocyclic group;

[0033] When the R is a substituted amino group, react the 9-bromo-6H-dibenzo[b,d]pyran-3-ol obtained in step S2 with an amino substituent to obtain 9-(arylamino)-6H-dibenzo[b,d]pyran-3-ol, then react 9-(arylamino)-6H-dibenzo[b,d]pyran-3-ol with methyl (R)-2-hydroxypropionate to obtain methyl (S)-2-[(9-(arylamino)-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate, and finally react methyl (S)-2-[(9-(arylamino)-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate with ammonia methanol to obtain a compound of general formula (I), a stereoisomer thereof, or a salt thereof, wherein R is a substituted amino group;

[0034] When the R is a heterocyclic group or a substituted heterocyclic group, react methyl (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate obtained in step S3 with a heterocyclic compound or a substituted heterocyclic compound to obtain methyl 9-(heterocyclic group or substituted heterocyclic group)-substituted (S)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propionate, and after reacting with ammonia methanol, obtain a compound of general formula (I), a stereoisomer thereof, or a salt thereof, wherein R is a heterocyclic group or a substituted heterocyclic group.

[0035] Compared with the prior art, the present invention has the following beneficial technical effects:

[0036] An (S)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide derivative, in which structures such as the 6H-dibenzo[b,d]pyran ring and the substituted heteroaryl ring at the 9-position in this derivative enhance the interaction with the hinge region and the affinity region of the catalytic active region of PI3Kα kinase respectively, and the 2-oxo group of propanamide at the 3-position interacts with the non-conserved amino acid Gln859, enhancing the specific binding to PI3Kα kinase, so that this derivative has better inhibitory activity against PI3Kα kinase and higher selectivity. Detailed implementation manners

[0037] To enable those skilled in the art to understand the features and effects of the present invention, the following provides a general description and definition of the terms and phrases mentioned in the specification and claims. Unless otherwise specified, all technical and scientific terms used herein have the ordinary meanings understood by those skilled in the art for the present invention. In case of conflicts, the definitions in this specification shall prevail. The theories or mechanisms described and disclosed herein, whether correct or incorrect, shall not limit the scope of the present invention in any way, that is, the content of the present invention can be implemented without being limited by any specific theory or mechanism. Herein, all features defined in the form of numerical ranges or percentage ranges, such as numerical values, quantities, contents and concentrations, are only for the sake of brevity and convenience. Accordingly, the description of numerical ranges or percentage ranges should be regarded as having covered and specifically disclosed all possible sub-ranges and individual numerical values within the ranges (including integers and fractions). Herein, unless otherwise specified, the terms "comprising", "including", "containing", "having" or similar terms cover the meanings of "consisting of" and "consisting essentially of". For example, "A comprises a" covers the meanings of "A comprises a and others" and "A only comprises a". Herein, for the sake of concise description, all possible combinations of all technical features in each embodiment or example are not described. Therefore, as long as there are no contradictions in the combinations of these technical features, the technical features in each embodiment or example can be combined arbitrarily, and all possible combinations should be considered as the scope described in this specification.

[0038] The present invention provides an (S)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide derivative, including the compound shown in general formula (I), the stereoisomer of general formula (I) or its salt. Preferably, it can be a pharmaceutically acceptable salt:

[0039]

[0040] Wherein, the R group is any one of heteroaryl, heterocyclic group, oxoheterocyclic group, thioheterocyclic group, substituted aryl, substituted heteroaryl, substituted heterocyclic group, substituted amino.

[0041] The present invention also provides a pharmaceutical composition, comprising the compound represented by general formula (I) in the present invention, the stereoisomer of general formula (I) or a salt thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients. The pharmaceutical composition is made into tablets, capsules, soft capsules or injections by adding excipients to the compound represented by general formula (I), the stereoisomer of general formula (I) or a salt thereof. Each tablet, capsule or injection contains 10-500 mg of the compound represented by general formula (I) in the present invention, the stereoisomer of general formula (I) or a salt thereof; the added excipients include one or several of additives, stabilizers, solubilizers, lubricants and disintegrants.

[0042] The present invention also provides the use of the compound represented by general formula (I) in the present invention, the stereoisomer of general formula (I) or a salt thereof, or the above-mentioned pharmaceutical composition in the preparation of PI3Kα inhibitor drugs or the preparation of anti-cancer drugs, wherein the cancer includes any one of breast cancer, pancreatic cancer, non-small cell lung cancer, thyroid cancer, seminoma, melanoma, bladder cancer, liver cancer, gastric cancer, kidney cancer, ovarian cancer, uterine cancer, prostate cancer, glioma, myelodysplastic syndrome, acute myeloid leukemia and colorectal cancer.

[0043] The structure of the compound of the present invention is determined by nuclear magnetic resonance (NMR) or / and gas chromatography-mass spectrometry (GC-MS) or high-resolution mass spectrometer. The NMR chemical shift (δ) is given in parts per million (ppm). The NMR measurement is carried out with a Bruker AVANCE-400 nuclear magnetic resonance instrument or an AVANCE III-600 nuclear magnetic resonance instrument. The solvents for measurement are deuterated dimethyl sulfoxide (DMSO-d6) and deuterated chloroform (CDCl3), and the internal standard is tetramethylsilane (TMS). The GC-MS measurement is carried out with a Shimadzu GCMS-QP2010 gas chromatography-mass spectrometer. The high-resolution mass spectrometry measurement is carried out with a Thermo Fisher Q Exactive Plus high-resolution mass spectrometer. The thin-layer chromatography silica gel plate uses a Qingdao GF254 silica gel plate. The TLC specification is 0.15 mm - 0.20 mm, and the specification for separating and purifying products by thin-layer chromatography is 0.4 mm - 0.5 mm. Column chromatography generally uses 200-300 mesh Yantai Yellow Sea silica gel as the carrier. The starting materials in the examples of the present invention are known and can be purchased on the market, or can be synthesized by methods known in the art. Without special instructions, all reactions of the present invention are carried out under continuous magnetic stirring in a dry nitrogen atmosphere, the solvent is a dry solvent, and the reaction temperature unit is degrees Celsius.

[0044] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. In addition, it should be understood that after reading the content taught by the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

[0045] In the following embodiments, conventional instrument equipment in the art is used. For the experimental methods without specific conditions noted in the following embodiments, they are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer. In the following embodiments, various raw materials are used. Unless otherwise stated, commercially available products are used, and their specifications are the conventional specifications in the art. In the specification of the present invention and the following embodiments, unless otherwise specified, "%" represents weight percentage, "parts" represents weight parts, and the ratio represents weight ratio.

[0046] A (S)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide derivative, including a compound represented by the general formula (I), a stereoisomer of the general formula (I), or a salt thereof:

[0047]

[0048] Among them, the R group is any one of a heteroaryl group, a heterocyclic group, an oxoheterocyclic group, a thioheterocyclic group, a substituted aryl group, a substituted heteroaryl group, a substituted heterocyclic group, and a substituted amino group.

[0049] That is, it includes the general formula

[0050]

[0051] Among them, R 1 is R 2 is the above-mentioned R group.

[0052] Table 1 shows the specific functional groups of the R group in a (S / R)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide derivative prepared in Examples 1 to 108 and the characterization results of the product. The corresponding Chinese names of the (S / R)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide derivatives prepared in Examples 1 to 108 are shown in Table 2.

[0053] Table 1 Characterization test data of the products in Examples 1 to 108

[0054]

[0055]

[0056]

[0057]

[0058]

[0059]

[0060]

[0061]

[0062]

[0063]

[0064]

[0065]

[0066] Among them, in Example 58 of Table 1, the R1 group is In the remaining examples, the R1 group is

[0067] Among them, the preparation method of the (S)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide derivative in Example 1, that is, (S)-2-[(9-(2-ethylpyridin-4-yl)-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide, is as follows:

[0068] The first step: Preparation of 3-hydroxy-9-bromo-6H-dibenzo[b,d]pyran-6-one

[0069]

[0070] Weigh 2,4-dibromobenzoic acid (12.00 g, 42.90 mmol), sodium hydroxide (3.24 g, 81.00 mmol) and resorcinol (9.36 g, 85.80 mmol) and place them in a three-necked flask. After protecting with nitrogen, slowly add 30 mL of water. After heating under reflux for 2 h, a large amount of yellow precipitate will precipitate. Subsequently, slowly drip 18 mL of 5% copper sulfate solution into the reaction system through a constant pressure dropping funnel. Immediately, a large amount of brick-red precipitate will precipitate. Continue heating under reflux for 1 h, stop heating, wait for the reaction system to cool to room temperature, filter by suction, obtain the brick-red product, discard the filtrate, and dry to obtain the title compound (8.75 g, 70%).

[0071] HRMS(ESI)[M-H] - : 288.9515. 11H NMR (600 MHz, DMSO-d6) δ 10.46 (s, 1H), 8.51 (s, 1H), 8.22 (d, J = 8.3 Hz, 1H), 8.06 (d, J = 8.4 Hz, 1H), 7.71 (d, J = 8.3 Hz, 1H), 6.83 (d, J = 8.7 Hz, 1H), 6.74 (s, 1H).

[0072] Step 2: Preparation of 9-bromo-6H-dibenzo[b,d]pyran-3-ol

[0073]

[0074] Weigh 3-hydroxy-9-bromo-6H-dibenzo[b,d]pyran-6-one (2.00 g, 6.87 mmol) and gallium tribromide (0.42 g, 1.37 mmol) and place them in a 100 mL three-necked flask. After nitrogen protection, add toluene (5 mL). Stir at room temperature, and slowly drip 1,1,3,3-tetramethyldisiloxane (1.95 mL, 15.80 mmol) into the three-necked flask. After the addition is complete, raise the temperature to 80 °C and continue the reaction for 15 minutes. Then, rotary evaporate the toluene under reduced pressure, extract three times with 20 mL of ethyl acetate / water, wash three times with saturated NaCl solution, and then combine the organic phases. The organic phases are dried over anhydrous sodium sulfate and then mixed with samples, and separated by silica gel column chromatography to obtain the title compound (1.24 g, 34%).

[0075] HRMS (ESI) [M-H]−: 274.9721. 1 1H NMR (600 MHz, DMSO-d6) δ 9.84 (s, 1H), 7.86 (s, 1H), 7.71 (d, J = 8.5 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.18 (d, J = 8.0 Hz, 1H), 6.50 (d, J = 8.5 Hz, 1H), 6.37 (s, 1H), 5.03 (d, J = 10.9 Hz, 2H).

[0076] Step 3: Preparation of methyl (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate

[0077]

[0078] Weigh 9-bromo-6H-dibenzo[b,d]pyran-3-ol (12.00 g, 43.20 mmol), methyl (R)-2-hydroxypropionate (6.72 g, 64.80 mmol) and triphenylphosphine (22.68 g, 86.40 mmol) and place them in a 100 mL three-necked flask. Measure 20 mL of 1,4-dioxane and add it to the flask. After ice-bathing for 10 min, place DIAD (17.52 g, 86.40 mmol) in a constant-pressure dropping funnel. After protecting with nitrogen, slowly drip DIAD into the three-necked flask. React at room temperature for 12 h. Monitor the reaction of the raw materials by TLC until it is complete. Evaporate the reaction solvent under reduced pressure. Extract with 50 mL of ethyl acetate / water three times. Wash with saturated NaCl solution three times and then combine the organic phases. Dry the organic phase over anhydrous sodium sulfate and then mix with the sample. Separate by silica gel column chromatography to obtain the title compound (14.07 g, 89%).

[0079] HRMS(ESI)[M+Na] + :385.0041. 1 H NMR(600MHz,DMSO-d6)δ7.93(s,1H),7.83(d,J=8.6Hz,

[0080] 1H),7.44(d,J=8.0Hz,1H),7.21(d,J=8.0Hz,1H),6.61(d,J=8.7Hz,1H),6.50(s,1H),5.08–5.04(m,3H),3.69(s,3H),1.51(d,J=6.6Hz,3H).

[0081] Step 4: Preparation of (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide

[0082]

[0083] Weigh methyl (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate (14.07 g, 38.74 mmol) and place it in a round-bottom flask. Add ammonia-methanol solution (140 mL, 7 mmol / mL) and react at room temperature for 12 h. Monitor the reaction of the raw materials by TLC until it is complete. Evaporate the solvent to obtain the title compound (14.05 g, 100%).

[0084] HRMS(ESI)[M+Na] + :370.0050. 1 H NMR(600MHz,DMSO-d6)δ7.93(s,1H),7.84(d,J=8.6Hz,

[0085] 1H), 7.55 (s, 1H), 7.43 (d, J = 8.0 Hz, 1H), 7.27 (s, 1H), 7.21 (d, J = 8.0 Hz, 1H), 6.64 (d, J = 8.7 Hz, 1H) 6.51 (s, 1H), 5.07 (s, 2H), 4.66 (q, J = 6.5 Hz, 1H), 1.44 (d, J = 6.5 Hz, 3H).

[0086] Step 5: Preparation of (S)-2-[(9-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide

[0087]

[0088] Weigh (S)-2-[(9-Bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide (2.50 g, 7.18 mmol), bis(pinacolato)diboron (2.70 g, 10.80 mmol), ferrocenedichloropalladium (0.30 g, 1.40 mmol) and anhydrous K2CO3 (2.00 g, 14.40 mmol), place them in a 100 mL three-necked flask. After protecting the reaction system with nitrogen, measure 60 mL of a liquid of 1,4-dioxane:water = 5:1 and inject it into the flask. Heat the mixture to 125 °C and react for 15 h. Monitor the reaction by TLC until the raw materials are completely reacted. Cool the reaction mixture to room temperature, extract it three times with 20 mL of ethyl acetate / water, wash it three times with saturated NaCl solution, then combine the organic phases. The organic phases are dried over anhydrous sodium sulfate and then mixed with silica gel. The title compound is obtained by silica gel column chromatography (2.39 g, 84%).

[0089] HRMS(ESI)[M+Na] + : 418.1798. 1 1H NMR (600 MHz, DMSO-d6) δ 7.92 (s, 1H), 7.86–7.69 (m, 1H), 7.62–7.49 (m, 2H), 7.26 (d, J = 7.7 Hz, 2H), 6.65 (d, J = 8.6 Hz, 1H), 6.51 (s, 1H), 5.12 (s, 2H), 4.64 (q, J = 6.5 Hz, 1H), 1.42 (dd, J = 24.8, 15.8 Hz, 3H), 1.31 (s, 12H).

[0090] Step 6: Preparation of (S)-2-[(9-(2-Ethylpyridin-4-yl)-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide

[0091]

[0092] Weigh (S)-2-[(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide (0.25 g, 0.63 mmol), 2-ethyl-4-bromopyridine (0.18 g, 0.95 mmol), ferrocenedichloropalladium (0.03 g, 0.13 mmol) and anhydrous K2CO3 (0.17 g, 1.26 mmol), place them in a 100 mL three-necked flask. After protecting the reaction system with nitrogen, measure 6 mL of a liquid of 1,4-dioxane:water = 5:1 and inject it into the flask. Heat the mixture to 125 °C and react for 15 h. Monitor the reaction by TLC. When the raw materials are basically completely reacted, extract with 20 mL of ethyl acetate / water three times, wash with saturated NaCl solution three times, then combine the organic phases. Dry the organic phases over anhydrous sodium sulfate and mix with samples. Separate by silica gel column chromatography to obtain the title compound (0.18 g, 76%).

[0093] HRMS(ESI)[M+H] + : 375.1711. 1 1H NMR(600MHz,DMSO-d6)δ8.54(d,J=5.0Hz,1H),8.13(s,

[0094] 1H),8.03(d,J=8.5Hz,1H),7.74–7.65(m,2H),7.62(d,J=4.9Hz,1H),7.56(s,1H),7.38(d,J=7.7Hz,1H),7.28(s,1H),6.69(d,J=8.7Hz,1H),6.54(s,1H),5.16(s,2H),4.68(d,J=6.6Hz,1H),2.83(q,J=7.6Hz,2H),1.45(d,J=6.4Hz,3H),1.29(t,J=7.5Hz,3H).

[0095] The preparation method of (S)-2-[(9-(2-oxazolidin-3-yl)-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide in Example 19 is as follows:

[0096] The first step: Preparation of methyl (S)-2-[(9-(2-oxazolidin-3-yl)-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate

[0097]

[0098] Weigh methyl (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate (0.26 g, 0.72 mmol), oxazolidin-2-one (0.13 g, 1.44 mmol), cuprous iodide (0.03 g, 0.14 mmol) and anhydrous K2CO3 (0.30 g, 2.16 mmol), place them in a 15 mL reaction tube, protect the reaction system with nitrogen. Take trans-N,N'-dimethyl-1,2-cyclohexanediamine (0.04 g, 0.29 mmol) and add it to 4 mL of toluene to mix, then inject the mixed solution into the reaction tube, heat up to 125 °C and react for 24 h. Monitor the reaction by TLC until the raw materials are completely reacted, cool down to room temperature to end the reaction, extract with 10 mL of ethyl acetate / water three times, wash with saturated NaCl solution three times, then combine the organic phases. The organic phases are dried over anhydrous sodium sulfate and then mixed with the sample, and separated by silica gel column chromatography to obtain a white solid (0.14 g, 53%).

[0099] Step 2: Preparation of (S)-2-[(9-(2-oxazolidin-3-yl)-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide

[0100]

[0101] Put all the obtained white solid into a flask, add 15 mL of ammonia methanol solution, stir at room temperature for 12 h to obtain the title compound (0.13 g, 100%).

[0102] HRMS(ESI)[M+Na] + : 377.1108. 1 H NMR(600 MHz, DMSO-d6)δ7.82(s, 1H), 7.75(d, J = 8.6 Hz,

[0103] 1H), 7.55(s, 1H), 7.51(d, J = 8.2 Hz, 1H), 7.26(d, J = 7.9 Hz, 2H), 6.68(dd, J = 8.6, 2.4 Hz, 1H), 6.52(d, J = 2.4 Hz, 1H), 5.08(s, 2H), 4.66(q, J = 6.6 Hz, 1H), 4.46(t, J = 8.0 Hz, 2H), 4.14(t, J = 8.0 Hz, 2H), 1.44(d, J = 6.6 Hz, 3H).

[0104] The preparation method of (S)-2-[(9-(phenylamino)-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide in Example 27 is as follows:

[0105] Step 1: Preparation of 9-(phenylamino)-6H-dibenzo[b,d]pyran-3-ol

[0106]

[0107] Weigh 9-bromo-6H-dibenzo[b,d]pyran-3-ol (0.35 g, 1.26 mmol), aniline (0.17 g, 1.44 mmol), BrettPhos G3 (6 mg, 0.0025 mmol) and sodium tert-butoxide (0.33 g, 3.15 mmol) into a 100 mL three-necked flask. Measure 6 mL of 1,4-dioxane and pour it into the flask. After protecting with nitrogen, heat the mixture to 110 °C and react for 12 h. Cool down to room temperature to end the reaction. Extract with 10 mL of ethyl acetate / water three times, wash with saturated NaCl solution three times, and then combine the organic phases. The organic phases are dried over anhydrous sodium sulfate, mixed with silica gel, and separated by silica gel column chromatography to obtain a white solid (0.27 g, 73%).

[0108] Step 2: Preparation of (S)-2-[(9-(phenylamino)-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide

[0109]

[0110] Weigh the white solid from the first step (0.12 g, 0.42 mmol), methyl (R)-2-hydroxypropionate (0.06 g, 0.54 mmol) and triphenylphosphine (0.17 g, 0.63 mmol) into a 100 mL three-necked flask. Measure 20 mL of 1,4-dioxane and pour it into the flask. After cooling in an ice bath for 10 min, place DIAD (0.13 g, 0.63 mmol) in a constant pressure dropping funnel. After protecting with nitrogen, slowly drop DIAD into the eggplant-shaped flask. React at room temperature for 12 h. Cool down to room temperature to end the reaction. Extract with 10 mL of ethyl acetate / water three times, wash with saturated NaCl solution three times, and then combine the organic phases. The organic phases are dried over anhydrous sodium sulfate, mixed with silica gel, and separated by silica gel column chromatography to obtain a white solid (0.08 g, 51%). Place all the obtained white solid in a flask, add 10 mL of ammonia-methanol solution, stir at room temperature for 12 h to obtain the title compound (0.08 g, 100%).

[0111] HRMS(ESI)[M+H] + :361.1549. 1 H NMR(600MHz,DMSO-d6)δ8.23(s,1H),7.60(d,J=8.4Hz,

[0112] 1H), 7.53 (s, 1H), 7.35 (s, 1H), 7.29–7.21 (m, 3H), 7.10 (t, J = 8.7 Hz, 3H), 6.97 (d, J = 8.0 Hz, 1H), 6.83 (t, J = 7.1 Hz, 1H), 6.63 (d, J = 8.5 Hz, 1H), 6.50 (s, 1H), 5.02 (s, 2H), 4.63 (d, J = 6.4 Hz, 1H), 1.43 (d, J = 6.3 Hz, 3H).

[0113] Table 2 Names of the products obtained in Examples 1 to 108

[0114]

[0115]

[0116]

[0117]

[0118] Among them, for the products corresponding to Examples 2 to 18 and 29 to 108, the preparation method refers to Example 1; for the products corresponding to Examples 20 to 26, the preparation method refers to Example 19; for the product corresponding to Example 28, the preparation method refers to Example 27.

[0119] That is, when the R is any one of heteroaryl, substituted aryl, substituted heteroaryl, oxoheterocyclic group or thioheterocyclic group, it includes the following steps:

[0120] S1: React 2,4-dibromobenzoic acid with resorcinol to obtain 3-hydroxy-9-bromo-6H-dibenzo[b,d]pyran-6-one;

[0121] S2: Reduce 3-hydroxy-9-bromo-6H-dibenzo[b,d]pyran-6-one by 1,1,3,3-tetramethyldisiloxane to obtain 9-bromo-6H-dibenzo[b,d]pyran-3-ol;

[0122] S3: React 9-bromo-6H-dibenzo[b,d]pyran-3-ol with methyl (R)-2-hydroxypropionate to prepare methyl (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate;

[0123] S4: React methyl (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate with a methanol solution of ammonia to prepare (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide;

[0124] S5: React (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide with bis(pinacolato)diboron to obtain (S)-2-[(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide;

[0125] S6: React (S)-2-[(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide with any one of a halogenated heteroaryl, a halogenated substituted aryl, a halogenated substituted heteroaryl, a halogenated oxoheterocyclic group, or a halogenated thioheterocyclic group to obtain a compound of general formula (I), a stereoisomer thereof, or a salt thereof, wherein R is any one of a heteroaryl, a substituted aryl, a substituted heteroaryl, an oxoheterocyclic group, or a thioheterocyclic group;

[0126] When the said R is a substituted amino group, react the 9-bromo-6H-dibenzo[b,d]pyran-3-ol obtained in the said step S2 with an amino substituent to obtain 9-(aryl amino)-6H-dibenzo[b,d]pyran-3-ol, then react 9-(aryl amino)-6H-dibenzo[b,d]pyran-3-ol with methyl (R)-2-hydroxypropionate to obtain methyl (S)-2-[(9-(aryl amino)-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate, and finally react methyl (S)-2-[(9-(aryl amino)-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate with ammonia in methanol to obtain a compound of general formula (I), a stereoisomer thereof, or a salt thereof, wherein R is a substituted amino group;

[0127] When the said R is a heterocyclic group or a substituted heterocyclic group, react methyl (S)-2-[(9-bromo-6H-dibenzo[b,d]pyran-3-yl)oxy]propionate obtained in the said step S3 with a heterocyclic compound or a substituted heterocyclic compound to obtain methyl 9-(heterocyclic group or substituted heterocyclic group)-substituted (S)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propionate, and after reacting with ammonia in methanol, obtain a compound of general formula (I), a stereoisomer thereof, or a salt thereof, wherein R is a heterocyclic group or a substituted heterocyclic group.

[0128] The results of the determination of the inhibitory effect of the (S)-2-[(6H-dibenzo[b,d]pyran-3-yl)oxy]propanamide derivatives prepared in Examples 1 to 108 of the present invention on PI3Kα / β / δ / γ kinases are shown in Table 3. The specific test procedure is as follows:

[0129] Experimental objective: To test the inhibitory activity of the compounds of the examples on PI3Kα / β / δ / γ kinases.

[0130] Experimental instruments: The centrifuge (5430) was purchased from Eppendorf, the pipette was purchased from Eppendorf, and the microplate reader (Envision, SN.1050214) was purchased from Perkin Elmer.

[0131] Experimental method: In this experiment, the ADP-GLO lipid kinase assay method of Promega (Promegaf#V9102) was used. The lipid kinases PI3Kα / β / δ / γ catalyze reactions in the presence of the substrate PIP2:3PS and ATP, generating ADP from ATP. The activity of the lipid kinase was characterized by measuring the content of ADP in the reaction, and the inhibition rate of the compound on the single concentration of the PI3Kα / β / δ / γ kinase activity, or the half-maximal inhibitory concentration IC 50 。

[0132] The specific experimental operations are as follows:

[0133] The kinase reaction was carried out in a white 384-well plate (Perkin Elmer#6007299). 50 nL of different concentrations of the compound at a 100-fold final concentration diluted with ddH2O containing 1% DMSO was added to each well. 50 nL of ddH2O containing 1% DMSO was added to the positive control well. Then, 2.5 μL of the PI3K kinase solution at a 2-fold final concentration diluted with 1× kinase buffer (HEPES 250 mM, MgCl2 15 mM, NaCl 250 mM, BSA 0.05%) was added to each well. 2.5 μL of 1× kinase buffer was added to the negative control well. After centrifugation at 1000 rpm for 30 seconds, the mixture was incubated at room temperature for 10 minutes after shaking and mixing evenly. 2.5 μL of a mixed solution of PIP2:3PS (Promega#V1701) and ATP at a 2-fold final concentration prepared with 1× kinase buffer was added to all wells to initiate the reaction. After centrifugation at 1000 rpm for 30 seconds and shaking and mixing evenly, the reaction was carried out at room temperature for 90 - 120 minutes. Then, 5 μL of ADP-Glo Reagent (containing 10 mM MgCl2) was added to each well and reacted at room temperature for 60 minutes to remove the excess ATP in the reaction. Then, 10 μL of Kinase Detection Reagent was added to each well, and the chemiluminescence value was detected with an Envision microplate reader after reacting in the dark at room temperature for 20 minutes.

[0134] Experimental data processing method:

[0135] Calculate the percentage inhibition data of the wells treated with the compound by using the positive control wells (DMSO control wells) and negative control wells (without adding kinase) on the plate {% inhibition rate = 100 - [(test compound value - negative control value) / (positive control value - negative control value)×100]}. For a series of concentrations, use GraphPad prism to fit the different concentrations and the corresponding percentage inhibition rate data to a four-parameter nonlinear logistic formula to calculate the IC 50 value.

[0136] Experimental conclusion:

[0137] Through the above protocol, the example compounds shown in the present invention showed the biological activities as shown in Tables 3 and 4 in the PI3Kα / β / δ / γ kinase activity assay.

[0138] Table 3 Inhibition rates of the products in Examples 1 to 108 and existing drugs on PI3Kα / β / δ / γ (%, concentration is 100 nmol / L)

[0139]

[0140]

[0141] Table 4 Inhibitory activity of the example compounds on PI3Kα / β / δ / γ and selectivity for PI3Kα (IC 50 , nmol / L)

[0142]

[0143]

[0144] As can be seen from Table 4, the example compounds shown in the present invention have good activity and selectivity in terms of PI3Kα / β / δ / γ kinase inhibition.

[0145] Furthermore, in order to verify the proliferation inhibitory effect of the example compounds of the present invention on PI3Kα mutant cancer cells, relevant tests were carried out. The specific test process is as follows:

[0146] Experimental purpose: The purpose of this example is to test the proliferation inhibitory activity of the example compounds on PI3Kα mutant breast cancer cells T-47D (H1047R) and MCF-7 (E545K).

[0147] Experimental instruments: The centrifuge (5430) was purchased from Eppendorf, the carbon dioxide incubator was purchased from Thermo, the biological safety cabinet was purchased from Shanghai Boxun, the pipette was purchased from Eppendorf, and the microplate reader (Envision, SN.1050214) was purchased from Perkin Elmer.

[0148] Experimental method: The CCK-8 method was used to detect the proliferation inhibitory effect of the tested compounds on PI3Kα mutant cancer cell lines (T-47D and MCF-7). MCF-7 cells were cultured in DMEM medium containing 10% FBS and 1% penicillin-streptomycin at 37 °C under 5% CO2. MCF-7 cells were seeded in 96-well plates at a density of 3000 cells / well and cultured overnight in an incubator at 37 °C under 5% CO2. After the cells adhered, different concentrations of the compound solution were added, and negative control and blank control were set simultaneously. Then, the cells were continuously cultured for 7 days at 37 °C under 5% CO2. Subsequently, 20 μL of CCK-8 reagent was added to each well, shaken and mixed evenly, and then cultured for 1 - 4 h. After the CCK-8 developed color, the 96-well plate was placed on an enzyme-linked immunosorbent assay (ELISA) reader, and the absorbance was measured at a wavelength of 450 nm to calculate the cell proliferation inhibition rate. T47D cells were cultured in RPMI-1640 medium containing 10% FBS, 1% penicillin-streptomycin and 0.01 mg / mL insulin at 37 °C under 5% CO2. T47D cells were seeded in 96-well plates at a density of 7500 cells / well and cultured overnight in an incubator at 37 °C under 5% CO2. After the cells adhered, different concentrations of the compound solution were added, and negative control and blank control were set simultaneously. Then, the cells were continuously cultured for 3 days at 37 °C under 5% CO2. Subsequently, 20 μL of CCK-8 reagent was added to each well, shaken and mixed evenly, and then cultured for 1 - 4 h. After the CCK-8 developed color, the 96-well plate was placed on an ELISA reader, and the absorbance was measured at a wavelength of 450 nm to calculate the cell proliferation inhibition rate.

[0149] Experimental data processing method: The percentage inhibition data of the wells treated with the tested compounds were calculated by comparing with the negative control wells on the plate {Inhibition rate (%) = 100 - (value of the tested compound - value of the blank control) / (value of the negative control - value of the blank control) × 100}. The IC 50 value was calculated by fitting the data of different concentrations and the corresponding percentage inhibition rates to a four-parameter non-linear logistic equation using GraphPad prism.

[0150] Experimental conclusion:

[0151] Through the above protocol, the tested compounds of the present invention showed the biological activities as shown in Table 5 in the proliferation inhibitory activity test on PI3Kα mutant breast cancer cells T-47D (H1047R) and MCF-7 (E545K).

[0152] Table 5 Proliferation inhibitory activity of the tested compounds on breast cancer cells (IC 50 μmol / L n = 3)

[0153]

[0154]

[0155] Furthermore, to verify the toxicity of the drug in the present invention, an acute toxicity experiment on mice (Experiment 1) was conducted. To verify the pharmacokinetic process of the drug, a pharmacokinetic experiment on rats (Experiment 2) was carried out, and at the same time, the in vivo efficacy test of the drug was explored (Experiment 3). The details are as follows:

[0156] Experiment 1: Acute toxicity experiment on mice

[0157] 1.1 Experimental purpose: The purpose of this study was to investigate the possible toxic reactions that might occur within 7 days after a single administration of Example 69 to Kunming mice by gavage, intraperitoneal injection, and tail vein injection.

[0158] 1.2 Experimental materials and instruments

[0159] Test article: Example 69; Solvent 1: 0.5% CMCNa aqueous solution (suspension); Solvent 2: 10% DMSO / 40% PEG300 / 5% Tween80 / 45% normal saline; Animal information: 120 Kunming mice (KM mice), half male and half female; Instruments: XW-80A vortex mixer; SB-120D ultrasonic cleaner

[0160] 1.3 Experimental method

[0161] 1) In the experiment, the mice were randomly divided into 4 groups by gender: Group 1 was the solvent control, Group 2 was given the drug by gavage, Group 3 was given the drug by intraperitoneal injection, and Group 4 was given the drug by tail vein injection;

[0162] 2) The animals in Group 1 were the control group, and each time they were given the blank solvent, which was administered simultaneously with the other three groups in the same way to observe whether the solvent had an impact on the mice;

[0163] 3) The animals in Group 2 were respectively given Example 69 by gavage at doses of 100, 200, and 400 mg / kg, with 5 males and 5 females at each dose;

[0164] 4) The animals in Group 3 were respectively given Example 69 by intraperitoneal injection at doses of 100, 200, and 400 mg / kg, with 5 males and 5 females at each dose;

[0165] 5) The animals in Group 4 were respectively given Example 69 by tail vein injection at doses of 50, 200, and 300 mg / kg, with 5 males and 5 females at each dose;

[0166] 6) Administration volume: 10 - 20 mL / kg by gavage; 10 mL / kg by intraperitoneal injection; 5 mL / kg by tail vein injection;

[0167] 7) Observe the status of mice during the experiment;

[0168] 8) Euthanize all animals after 7 days.

[0169] 1.4 Experimental data results

[0170] 1.4.1 Blank group

[0171] No abnormal symptoms were found in the blank solvent group, and no deaths / near-deaths were seen.

[0172] 1.4.2 Gavage group

[0173] No deaths / near-deaths were seen in the mice of the gavage administration group. After gavaging with Example 69 at 100 mg / kg, the mice showed slightly dull reactions within 1 h. After gavaging with 200 and 400 mg / kg, abnormal symptoms such as arched backs, fluffy hair, and dull reactions were seen within 2 h, and all symptoms subsided and returned to normal after 24 h.

[0174] 1.4.3 Intraperitoneal injection group

[0175] No deaths / near-deaths were seen in the mice of the intraperitoneal injection group. After intraperitoneal injection of Example 69 at 100, 200, and 400 mg / kg, abnormal symptoms such as arched backs, fluffy hair, and dull reactions were seen, and they became more severe with the increase of the administration dose, and the duration also increased. All symptoms subsided and returned to normal after 24 h.

[0176] 1.4.4 Tail vein injection group

[0177] After tail vein injection of Example 69 at 300 mg / kg, the mice died immediately. After tail vein injection of Example 69 at 50 and 200 mg / kg, abnormal symptoms such as arched backs, fluffy hair, and dull reactions were seen, and they became more severe with the increase of the administration dose, and the duration also increased. All symptoms subsided and returned to normal after 24 h.

[0178] 1.4 Experimental conclusion

[0179] Under the conditions of this experiment, the maximum tolerated dose of a single gavage of Example 69 in mice is greater than 400 mg / kg, the maximum tolerated dose of a single intraperitoneal injection is greater than 400 mg / kg, and the maximum tolerated dose of a single tail vein injection is between 200 - 300 mg / kg.

[0180] Experiment 2: Pharmacokinetics experiment of rats

[0181] 2.1 Experimental method

[0182] Animal information: 12 Sprague-Dawley (SD) rats; Administration method: single-dose gavage administration, single-dose intravenous injection; Test article: Example 69; Administration dose: 7.5 mg / kg by gavage, 3 mg / kg by intravenous injection; Administration volume: 5 mL / kg by gavage, 0.5 mL / kg by intravenous injection; Internal standard compound: Nimodipine; Formulation prescription 1: 0.5% CMCNa, ultrasonic treatment, prepared into a uniform suspension; Formulation prescription 2: 10% DMSO / 40% PEG300 / 5% Tween80 / 45% normal saline; Sampling points: 0.083, 0.25, 0.5, 1, 2, 3, 5, 7, 11, and 24 hours after administration.

[0183] Sample preparation:

[0184] 1) Collect 0.3 mL of jugular vein blood and place it in a centrifuge tube containing sodium heparin. Centrifuge at 12000 r / min for 5 min at room temperature to separate plasma, and store it at -20 °C.

[0185] 2) Take 100 μL of the separated plasma, add 1.0 mL of acetonitrile and 100 μL of the internal standard solution, vortex mix, centrifuge at 12000 r / min for 5 min after mixing, take the supernatant and dry it under nitrogen in a centrifuge tube, then add 200 μL of methanol, vortex mix for 5 min and centrifuge at 12000 r / min for 5 min.

[0186] 3) Take 100 μL of the treated supernatant solution for LC / MS / MS analysis to determine the concentration of the test example.

[0187] 2.2 HPLC-MS / MS determination conditions

[0188] Chromatographic column: Shim-pack GIST HP-C18 (2.1×100 mm, 3 μm); Column temperature: 35 °C; Mobile phase A: ultrapure water; Mobile phase B: methanol solution; Flow rate: 0.200 mL / min; Injection volume: 5.0 μL; Mass spectrometry conditions are shown in Table 6:

[0189] Table 6 Mass spectrometry detection conditions for Example 69 and Nimodipine (internal standard)

[0190]

[0191] 2.3 Pharmacokinetic results:

[0192] The main parameters were calculated using WinNonlin 8.1.0, and the results of the rat pharmacokinetic experiment are shown in Table 7:

[0193] Table 7 Results of the pharmacokinetic experiment of Example 69 in rats by gavage

[0194]

[0195] The results of the pharmacokinetic experiments on rats in the table show that Example 69 of the present invention is rapidly absorbed and eliminated in rats. The absolute bioavailability of Example 69 administered by gavage to rats is 37.2% (7.5 mg). Both the plasma exposure AUC and the maximum blood drug concentration Cmax showed good performance.

[0196] Experiment 3: In vivo pharmacodynamic experiment

[0197] 3.1 Experimental purpose:

[0198] To test the pharmacodynamic effect of Example 69 in vivo and to test the toxic and side effects of long-term drug administration.

[0199] 3.2 Experimental instruments and materials:

[0200] 3.2.1 Instruments:

[0201] 1. Biological safety cabinet (BSC-1300IIA2, Shanghai Boxun Industrial Co., Ltd., Medical Equipment Factory); 2. Ultra-clean workbench (CJ-2F, Suzhou Feng's Experimental Animal Equipment Co., Ltd.); 3. CO2 incubator (Thermo-311); 4. Centrifuge (Centrifuge 5702R, Eppendorf); 5. Automatic cell counter (CountessII, Life); 6. Pipette (Eppendorf); 7. Microscope (TS2, Nikon); 8. Vernier caliper (CD-6AX, Mitutoyo, Japan); 9. Cell culture flask (T75 / T225, Corning); 10. Electronic balance (CPA2202S, Sartorius)

[0202] 3.2.2 Reagents:

[0203] 1. DMEM medium; 2. Fetal bovine serum (FBS); 3. 0.25% trypsin; 4. Phosphate buffer solution (PBS); 5. Matrigel Matrix

[0204] 3.2.3 Animals:

[0205] NCG mice, 4-6 weeks old, ♀, purchased from Chengdu Jicui Yakang Co., Ltd. Three days before cell inoculation, estrogen was added to the drinking water to a final concentration of 2.0 μmol / L, and the drinking water was changed every three days.

[0206] 3.3 Experimental procedures:

[0207] 3.3.1 Cell culture and cell suspension preparation

[0208] a. Take out a strain of MCF7 cells from the cell bank, resuscitate the cells with DMEM medium, and place the resuscitated cells in a cell culture flask and culture them in a CO2 incubator (the temperature of the incubator is 37 °C and the CO2 concentration is 5%);

[0209] b. After the cells cover 80 - 90% of the bottom of the culture flask, passage the cells, and continue to culture the cells in the CO2 incubator after passage. Repeat this process until the number of cells meets the in vivo efficacy requirements;

[0210] c. Collect the cultured cells, count them with an automatic cell counter, and resuspend the cells with PBS and Matrigel according to the counting results to prepare a cell suspension, and place it on an ice box for later use.

[0211] 3.3.2 Cell seeding

[0212] a. Before seeding, label the NCG mice with disposable universal ear tags for mice and rats;

[0213] b. When seeding, mix the cell suspension evenly, draw the cell suspension with a 1 mL syringe, remove the air bubbles, and then place the syringe on an ice bag for later use;

[0214] c. Fix the NCG mouse with the left hand, depilate the area near the right thigh inguinal region (seeding site) of the mouse with depilatory cream, then disinfect it with 75% alcohol, and start seeding after 30 seconds;

[0215] d. Seed the test mice in turn (each mouse is seeded with 0.1 mL of cell suspension, and the number of seeded cells is 5×10 6 cells).

[0216] 3.3.3 Tumor measurement, grouping, and drug administration for tumor-bearing mice

[0217] a. According to the tumor growth situation, measure the tumor and calculate the tumor size on the 14th - 21st day after seeding. Tumor volume calculation: Tumor volume (mm3) = length (mm) × width (mm) × width (mm) × 0.52;

[0218] b. According to the body weight and tumor size of the tumor-bearing mice, use the method of random grouping for grouping;

[0219] c. According to the grouping results, start to administer the test drug (administration method: gavage; administration dose: 12.5 mg / kg; administration volume: 10 mL / kg; administration frequency: once a day; administration cycle: 21 days; solvent: 0.5% CMCNa);

[0220] d. Measure the tumor and weigh the mice once every three days after starting to administer the test drug;

[0221] e. Euthanize the animals after the experiment ends.

[0222] f. Process the data using software such as Excel. Calculation of the tumor growth inhibition rate TGI (%) of the compound: When the tumor does not regress, TGI (%) = [(1 - (average tumor volume at the end of drug administration in the treatment group - average tumor volume at the start of drug administration in the treatment group) / (average tumor volume at the end of treatment in the solvent control group - average tumor volume at the start of treatment in the solvent control group))] × 100%. When the tumor regresses, TGI (%) = [1 - (average tumor volume at the end of drug administration in the treatment group - average tumor volume at the start of drug administration in the treatment group) / average tumor volume at the start of drug administration in the treatment group] × 100%.

[0223] 3.4 The test data are shown in Table 8:

[0224] Table 8 Tumor growth inhibition rate of intragastric administration of Example 69 to tumor-bearing NCG mice

[0225]

[0226] During the drug administration period, the body weights of the mice in the solvent control group and the intragastric administration of Example 69 group increased slowly, and there was no significant difference in the body weights of the two groups of mice.

[0227] 3.4 Test conclusion:

[0228] It can be seen from the above results that Example 69 shows strong in vivo anti-tumor activity. Example 69 has no effect on the body weight of mice, indicating that Example 69 has good tolerance and good safety.

[0229] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

[0230] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the protection scope of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A ([[$ S )-2-( 6H -dibenz b, d pyran-3-yl)oxy]propanamide derivative, characterized in that, Comprising a compound represented by general formula (I), a stereoisomer thereof or a salt thereof: Wherein, the R group is: ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and any one of them.

2. A pharmaceutical composition, characterized in that, Comprising the compound represented by general formula (I), the stereoisomer thereof or a salt thereof as described in claim 1, and one or more pharmaceutically acceptable carriers, diluents or excipients; the pharmaceutical composition is made into tablets, capsules or injections by adding excipients to the compound, its stereoisomer or a salt thereof as shown in claim 1; the excipients include one or several of stabilizers, solubilizers, lubricants and disintegrants.

3. Use of the compound represented by general formula (I), the stereoisomer thereof or a salt thereof as described in claim 1, or the pharmaceutical composition of claim 2 in the preparation of a PI3Kα inhibitor drug.

4. Use of the compound represented by the general formula (I), stereoisomers of the general formula (I), or salts thereof as described in claim 1, or the pharmaceutical composition in claim 2, in the preparation of a cancer drug; characterized in that, The cancer includes any one of breast cancer, pancreatic cancer, non-small cell lung cancer, thyroid cancer, seminoma, melanoma, bladder cancer, liver cancer, gastric cancer, kidney cancer, ovarian cancer, uterine cancer, prostate cancer, glioma, myelodysplastic syndrome, acute myeloid leukemia and colorectal cancer.

5. Preparation method of the compound represented by general formula (I), the stereoisomer thereof or a salt thereof as described in any one of claim 1, characterized in that When the R is 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 , , , , , , , , , , , , , , , , , , , , , or when, the following steps are included: S1: React 2,4-dibromobenzoic acid with resorcinol to obtain 3-hydroxy-9-bromo-6 H -dibenz b, d pyran-6-one; S2: 3-Hydroxy-9-bromo-6 H -dibenz b, d pyran-6-one is reduced by 1,1,3,3-tetramethyldisiloxane to obtain 9-bromo-6 H -dibenz b, d pyran-3-ol; S3: React 9-bromo-6 H -dibenz b, d pyran-3-ol with methyl ([[]] R )-2-hydroxypropionate to obtain methyl ([[]] S )-2-[(9-bromo-6 H -dibenz b, d pyran-3-yl)oxy]propionate; S4: React the methyl ([ S ])-2-[(9-bromo-6 H -dibenz[[ b, d ]]pyran-3-yl)oxy]propionate with an ammonia methanol solution to obtain ([ S ])-2-[(9-bromo- 6H -dibenz[[ b, d ]]pyran-3-yl)oxy]propanamide;​​​​​​​​​​​​ S5: React ( S )-2-[(9-bromo- 6H -dibenz b, d pyran-3-yl)oxy]propanamide with bis(pinacolato)diboron to obtain ( S )-2-[(9-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)- 6H -dibenz b, d pyran-3-yl)oxy]propanamide; S6: Reacting ([ S )-2-[(9-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-6 H -dibenzo[[ b, d pyran-3-yl)oxy]propanamide with any one of a halogenated heteroaryl, a halogenated substituted aryl, and a halogenated substituted heteroaryl to obtain a compound of general formula (I), a stereoisomer thereof, or a salt thereof; When the R is or , the 9-bromo-6 H -dibenz b, d pyran-3-ol obtained in the step S2 reacts with an amino substituent to obtain 9-(aryl amino)- 6H -dibenz b, d pyran-3-ol, and then 9-(aryl amino)- 6H -dibenz b, d pyran-3-ol reacts with methyl ([[]] R )-2-hydroxypropionate to obtain methyl ([[]] S )-2-[(9-(aryl amino)-6 H -dibenz[b,d]pyran-3-yl)oxy]propionate, and finally methyl ([[]] S )-2-[(9-(aryl amino)-6 H -dibenz[b,d]pyran-3-yl)oxy]propionate reacts with ammonia in methanol to prepare a compound of the general formula (I), a stereoisomer thereof or a salt thereof; When the R is , , , , , , or , react one of methyl ( S )-2-[(9-bromo-6 H -dibenzo[b,d]pyran-3-yl)oxy]propionate obtained in the step S3 with a heterocyclic compound or a substituted heterocyclic compound to prepare methyl 9-heterocyclic group or substituted heterocyclic group substituted ( S )-2-[(6 H -dibenzo[b,d]pyran-3-yl)oxy]propionate. After reacting with ammonia methanol, a compound of the general formula (I), a stereoisomer thereof or a salt thereof is prepared.

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