A polysubstituted coumarin compound and applications

Abstract

The present application provides a polysubstituted coumarin compound having the general structure of (I) and its pharmaceutically acceptable salts, solvates and uses, wherein R1 is defined herein. The present application also relates to the preparation of the compound, pharmaceutical compositions of the compound and the use of the compound for treating related diseases by inhibiting inositol requiring enzyme 1 alpha (IRE1 alpha), such as for the treatment of diseases such as cancer and pulmonary fibrosis.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and in particular relates to a polysubstituted coumarin compound and its application and preparation method. Background Art

[0002] In recent years, many studies have found that endoplasmic reticulum stress (ERS) plays an important role in the occurrence and development of diseases such as cancer and pulmonary fibrosis. The endoplasmic reticulum (ER) is a special organelle located in eukaryotic cells. It plays an important role in regulating biological processes such as protein biosynthesis, folding, lipid biosynthesis, cell apoptosis and calcium homeostasis (Shi Z, Xu L, Xie H, et al. BMC Pulmonary Medicine, 2020, 20(1):92). Whether the internal environment of the ER is stable plays an important role in its normal physiological function and is also an important basis for the stability of the entire cell's internal environment. Any factor that interferes with the stability of the internal environment of the ER may cause misfolded proteins to accumulate in the ER, a situation known as ERS. The perception and response of ERS reactions are regulated by the unfolded protein response (UPR), an integrated adaptive pathway that evolved to maintain endoplasmic reticulum protein homeostasis (Salvagno C, Mandula JK, Rodriguez PC, et al. Trends in Cancer, 2022, 8(11):930-943). When ERS occurs, the adaptive mechanism of the UPR is triggered, allowing cells to restore homeostasis and restore the endoplasmic reticulum to a normal physiological state. However, in cases of persistent or severe ERS, the UPR's ability to restore cells is broken, leading to cell death. The UPR is a complex signaling pathway that is mainly mediated by three effector pathways, including the activation of PKR-like endoplasmic reticulum kinase (PERK), transcription activator 6 (ATF6) and inositol-requiring enzyme 1α (IRE1α) (Nanthakumar CB, Hatley RJD, Lemma S, et al. Nature Reviews Drug Discovery. 2015, 14(10):693–720). Glucose-regulated protein 78 (Grp78), also known as immunoglobulin heavy chain binding protein (Bip), is a member of the heat shock protein family and is crucial for UPR regulation. Under normal circumstances, Grp78 / Bip binds to three endoplasmic reticulum proteins (PERK, ATF6, and IRE1α) to form an inactive complex and inhibit their signaling. Grp78 / Bip also binds to misfolded proteins in the endoplasmic reticulum. Therefore, as misfolded proteins accumulate, Grp78 / Bip dissociates from these three proteins, PERK, ATF6, and IRE1α, activating the UPR to improve protein folding and maintain the stability of the intracellular environment.However, if ERS persists and worsens, PERK, ATF6 and IRE1α will induce and activate corresponding signaling pathways, leading to cell apoptosis (Bertolotti A, Zhang Y, Hendershot LM, et al. Nature Cell Biology, 2000, 2(6): 326–332).

[0003] IRE1α is a bifunctional enzyme with endonuclease and kinase activities located in the endoplasmic reticulum membrane. After being activated by ERS, IRE1α polymerizes and autophosphorylates, activating its endoribonuclease activity, removing the intron of the transcription factor XBP1 and converting it into the active form XBP1s. XBP1s then translocates to the nucleus and promotes the transcription of components of the ERAD system, including the endoplasmic reticulum degradation-enhancing α-mannosidase-like protein (Yoshida H, Matsui T, Yamamoto A, et al. Cell, 2001, 107(7):881–891). Activated IRE1α can also oligomerize and autophosphorylate, reducing the endoribonuclease activity of oligomeric IRE1α and promoting the degradation of mRNA and microRNA (miRNA) through IRE1α-dependent decay (Hollien J, Lin JH, Li H, et al. Journal of Cell Biology, 2009, 186(3):323–331). PERK, ATF6, and IRE1α signaling proteins work together to maintain cellular homeostasis in the ERS environment.

[0004] ORIN1001 is an IRE1α inhibitor developed by Fosun Hongchuang (Suzhou) Pharmaceutical Technology Co., Ltd. It has shown good therapeutic effects on idiopathic pulmonary fibrosis and metastatic breast cancer in phase I and II clinical trials.

[0005] In summary, IRE1α inhibitors play an important role in the treatment of diseases such as cancer and pulmonary fibrosis. Therefore, it is urgent to find and discover new IRE1α inhibitors. Summary of the Invention

[0006] To address these issues, the present invention proposes a polysubstituted coumarin compound and its use. This compound exhibits favorable pharmacological properties, inhibiting IRE1α and demonstrating superior antiproliferative activity against Mlg2908 and H358 cells compared to ORIN1001, currently in Phase II clinical trials. Therefore, the compound of formula (I) is suitable for treating IRE1α-dependent diseases, particularly proliferative diseases such as cancer and pulmonary fibrosis.

[0007] In the first aspect, the present invention provides a polysubstituted coumarin compound, the structural formula of which is as follows:

[0008]

[0009] Wherein, R1 is selected from

[0010]

[0011]

[0012] More specifically, the structural formula of the polysubstituted coumarin compound is one of the following:

[0013]

[0014]

[0015]

[0016] In a second aspect, the present invention provides a use of a polysubstituted coumarin compound as a drug for treating diseases mediated by inositol-requiring enzyme 1α (IRE1α).

[0017] Furthermore, the disease mediated by the inositol-requiring enzyme 1α (IRE1α) is cancer or pulmonary fibrosis; the cancer is one of lung and bronchial cancer, prostate cancer, breast cancer, pancreatic cancer, colon and rectal cancer, thyroid cancer, liver and intrahepatic bile duct cancer, hepatocellular carcinoma, gastric cancer, glioma / glioblastoma, endometrial cancer, melanoma, kidney and renal pelvis cancer, bladder cancer, uterine body cancer, cervical cancer, ovarian cancer, multiple myeloma, esophageal cancer, acute myeloid leukemia, chronic myeloid leukemia, lymphocytic leukemia, myeloid leukemia, brain cancer, oral and pharyngeal cancer, laryngeal cancer, small intestine cancer, non-Hodgkin's lymphoma, melanoma and villous colon adenoma.

[0018] In a third aspect, the present invention provides a pharmaceutical composition comprising the polysubstituted coumarin compound according to claim 1 and a pharmaceutically acceptable excipient.

[0019] The polysubstituted coumarin compounds having the general structural formula (I) provided by the present invention are prepared by the following steps, but are not limited to the following methods:

[0020]

[0021] wherein R1 is as described in claim 1.

[0022] The specific reaction process is as follows:

[0023] Synthesis of compound of formula (III): To a suspension of 4-methoxyresorcinol (1.0 eq) in acetylsuccinyl diethyl ester (1.2 eq) was added CH3SO3H. The mixture was stirred at room temperature for 12 h. The organic layer was extracted with ethyl acetate, anhydrous sodium sulfate was used to remove water, and the solvent was removed under reduced pressure to obtain a crude product, which was further purified by silica gel column separation to obtain the compound of formula (III).

[0024] Synthesis of compound of formula (IV): The compound of formula (III) was added to a 4 mol / L NaOH solution at one time and stirred at room temperature for 4 h. The reaction mixture was acidified with 5 mol / L HCl to pH = 3. The precipitate formed was filtered, filtered with ethyl acetate and dried under vacuum to obtain the compound of formula (IV).

[0025] Synthesis of compound of formula (V): The compound of formula (IV) (1.0 eq) and HATU (1.5 eq) were dissolved in anhydrous DMF, and DIPEA (4.0 eq) and an amine-containing compound R1 (1.2 eq) were added in sequence. The mixture was stirred at room temperature for 3-5 h. The organic layer was extracted with ethyl acetate, anhydrous sodium sulfate was used to remove water, and further purified by silica gel column separation to obtain the compound of formula (V).

[0026] Synthesis of compound of formula (I): A mixture of the compound of formula (V) (1.0 eq) and HMTA (3.0-4.0 eq) in TFA was heated at 90-120°C under nitrogen for 1.5-3 h. The organic layer was extracted with dichloromethane, anhydrous sodium sulfate was used to remove water, and further purified by silica gel column separation to obtain the compound of formula (I).

[0027] The present application has the beneficial effect of having favorable pharmacological properties, being able to inhibit IRE1α, and having a better inhibitory activity on the proliferation of Mlg2908 cells and H358 cells than ORIN1001 in clinical phase II. DETAILED DESCRIPTION

[0028] The specific examples contained herein are for purposes of illustration only and should not be construed as limiting the scope of the application. Further, those skilled in the art will appreciate that various modifications can be made to the application herein described and that such modifications come within the scope of the appended claims.

[0029] The synthesis route of the polysubstituted coumarin compound is as follows:

[0030]

[0031] Example 1: Preparation of (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2- oxoethyl)-2-oxo-2H-chromen-8-carboxaldehyde (Compound I-1)

[0032] Step 1: Preparation of ethyl 2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetate (Compound III)

[0033]

[0034] To a suspension of 4-methoxyresorcinol (18.0 g, 258 mmol, 1.0 eq) in diethyl acetyl succinate (30.6 mL, 309 mmol, 1.2 eq) was added CH₃SO₃H₄. The mixture was stirred at room temperature for 12 h. The organic layer was extracted with ethyl acetate, and the water was removed by anhydrous sodium sulfate. The solvent was then distilled off under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography to obtain 60.3 g of a white solid (Compound III). The yield was 80.0%. 1 H NMR(400MHz,DMSO-d6)δ10.22(s,1H),7.16(s,1H),6.75(s,1H),4.05(d,J=7.2Hz,2H), 3.84(s,3H),3.60(s,2H),2.35(s,3H),1.15(t,J=7.2Hz,3H); ESI-MS:m / z=293.10[M+H] + .

[0035] Step 2: Preparation of 2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetic acid (Compound IV)

[0036]

[0037] Ethyl 2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetate (60.0 g, 205 mmol) was added all at once to 30 mL of 4 mol / L NaOH solution and stirred at room temperature for 4 h. The reaction mixture was acidified to pH 3 with 5 mol / L HCl. The resulting precipitate was filtered, eluted with ethyl acetate, and dried under vacuum to yield 39.4 g of a pale pink solid (Compound IV). Yield: 72.8%. 1 H NMR (400MHz, DMSO-d6) δ12.39(s,1H),10.22(s,1H),7.17(s,1H),6.77(s,1H),3.85(s,3H),3.54(s,2H),2.35(s,3H); ESI-MS: m / z=265.07[M+H] + .

[0038] Step 3: Preparation of (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3- methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (Compound V-1)

[0039]

[0040] A mixture of (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2- oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) and HMTA (145.2 mg, 1.0364 mmol, 3.0 eq) in TFA was heated at 90 °C under nitrogen for 1.5 h. The organic layer was extracted by dichloromethane, and the water was removed by anhydrous sodium sulfate, and then purified by silica gel column to give yellow-green solid (Compound I-1) 40.0 mg. Yield 30.9%. 1 H NMR (400 MHz, DMSO-d6) δ 10.16 (s, 1H), 7.14 (s, 1H), 6.75 (s, 1H), 4.20 (d, J = 60.3 Hz, 2H), 3.84 (s, 3H), 3.73-3.38 (m, 7H), 2.28 (s, 3H), 1.30 (s, 3H); ESI-MS: m / z = 348.14 [M+H] + .

[0041] Step 4: Preparation of (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3- methylmorpholino)-2-oxoethyl)-2-oxo-2H-chromene-8-carbaldehyde (Compound I-1)

[0042]

[0043] A mixture of (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2- oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) and HMTA (145.2 mg, 1.0364 mmol, 3.0 eq) in TFA was heated at 90 °C under nitrogen for 1.5 h. The organic layer was extracted by dichloromethane, and the water was removed by anhydrous sodium sulfate, and then purified by silica gel column to give yellow-green solid (Compound I-1) 40.0 mg. Yield 30.9%. 1H NMR (400MHz, DMSO-d6) δ11.81(s,1H),10.40(s,1H),7.42(s,1H),4.24(d,J=59.2Hz,1H),3.91( s,3H),3.88-3.33(m,8H),2.33(s,3H),1.24(dd,J=87.4,5.2Hz,3H); ESI-MS:m / z=398.12[M+Na] + .

[0044] Example 2: Preparation of (S)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2-oxo-2H-chromene-8-carbaldehyde (Compound I-2)

[0045] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (S)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) to obtain 256.8 mg of an off-white solid ((S)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one) in a yield of 79.0%. 1 HNMR(400MHz,DMSO-d6)δ10.16(s,1H),7.13(s,1H),6.74(s,1H),4.20(d,J=62.4Hz,2H),3. 83(s,3H),3.76-3.34(m,7H),2.27(s,3H),1.25(d,J=42.2Hz,3H); ESI-MS:m / z=348.14[M+H] + .

[0046]

[0047] In step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) was replaced with (S)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) to give 32.5 mg of a yellow-green solid (compound I-2) in a yield of 25.1%. 1H NMR (400MHz, DMSO-d6) δ11.74(s,1H),10.36(s,1H),7.37(s,1H),4.21(d,J=61.6Hz,1H),3.87( s,3H),3.82-3.22(m,8H),2.29(s,3H),1.21(dd,J=87.2,4.8Hz,3H); ESI-MS:m / z=398.12[M+Na] + .

[0048] Example 3: Preparation of (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(2-methylmorpholino)-2-oxoethyl)-2-oxo-2H-chromene-8-carbaldehyde (Compound I-3)

[0049] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (R)-2-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) to obtain 233.7 mg of an off-white solid ((R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(2-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one) in a yield of 71.9%. 1 HNMR(400MHz,DMSO-d6)δ10.16(s,1H),7.14(s,1H),6.74(s,1H),4.20-4.01(m,2H),3.83 (s,3H),3.81-3.37(m,7H),2.28(s,3H),1.08(d,J=14.0Hz,3H); ESI-MS:m / z=348.14[M+H] + .

[0050]

[0051] In step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) was replaced with (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(2-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) to give 35.7 mg of a yellow-green solid (compound I-3) in a yield of 27.6%. 1H NMR (400MHz, DMSO-d6) δ11.67(s,1H),10.26(s,1H),7.26(s,1H),4.10-3.92(m,1H),3.76( s,3H),3.69-2.67(m,8H),2.19(s,3H),0.98(d,J=20.6Hz,3H); ESI-MS:m / z=398.12[M+Na] + .

[0052] Example 4: Preparation of (S)-7-hydroxy-6-methoxy-4-methyl-3-(2-(2-methylmorpholino)-2-oxoethyl)-2-oxo-2H-chromene-8-carbaldehyde (Compound I-4)

[0053] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (S)-2-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) to obtain 249.6 mg of an off-white solid ((S)-7-hydroxy-6-methoxy-4-methyl-3-(2-(2-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one) in a yield of 76.8%. 1 HNMR(400MHz,DMSO-d6)δ10.17(s,1H),7.13(s,1H),6.74(s,1H),4.17-3.93(m,2H),3.83(s, 3H), 3.81-3.41 (m, 7H), 2.28 (s, 3H), 1.08 (dd, J=18.7, 5.7Hz, 3H); ESI-MS: m / z=348.14[M+H] + .

[0054]

[0055] In step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) was replaced with (S)-7-hydroxy-6-methoxy-4-methyl-3-(2-(2-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) to give 28.7 mg of a yellow-green solid (compound I-4) in a yield of 22.2%. 1H NMR(400MHz,DMSO-d6)δ11.79(s,1H),10.31(s,1H),7.32(s,1H),4.04(s,1H),3.81(s, 3H), 3.76-2.76 (m, 8H), 2.24 (s, 3H), 1.02 (d, J=19.6Hz, 3H); ESI-MS: m / z=398.12[M+Na] + .

[0056] Example 5: Preparation of 3-(2-((2S,6S)-2,6-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromene-8-carbaldehyde (Compound I-5)

[0057] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (2S,6S)-2,6-dimethylmorpholine (129.3 mg, 1.1240 mmol, 1.2 eq) to obtain 270.8 mg of an off-white solid (3-(2-((2S,6S)-2,6-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one) in a yield of 80.1%. 1 H NMR (400MHz, DMSO-d6) δ10.16(s,1H),7.14(s,1H),6.74(s,1H),3.95(s,1H),3.83(s,3H),3.72-3. 43(m,5H),3.29-3.14(m,2H),2.28(s,3H),1.08(dd,J=37.6,6.0Hz,6H); ESI-MS:m / z=362.16[M+H] + .

[0058]

[0059] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced with 3-(2-((2S,6S)-2,6-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one (124.7 mg, 0.3454 mmol, 1.0 eq) to obtain 31.2 mg of a yellow-green solid (compound I-5), with a yield of 23.1%. 1H NMR(400MHz,DMSO-d6)δ11.71(s,1H),10.21(s,1H),7.22(s,1H),3.82(s,1H),3.72(s, 3H), 3.61-3.00 (m, 7H), 2.14 (s, 3H), 0.94 (d, J=38.8Hz, 6H); ESI-MS: m / z=412.13[M+Na] + .

[0060] Example 6: Preparation of 3-(2-((2R,6R)-2,6-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromene-8-carbaldehyde (Compound I-6)

[0061] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (2R,6R)-2,6-dimethylmorpholine (129.3 mg, 1.1240 mmol, 1.2 eq) to obtain 236.8 mg of an off-white solid (3-(2-((2R,6R)-2,6-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one) in a yield of 70.0%. 1 HNMR(400MHz,DMSO-d6)δ10.20(s,1H),7.13(s,1H),6.76(s,1H),3.95(s,1H),3.83(s,3H),3.73-3 .39(m,5H),3.29-3.14(m,2H),2.28(s,3H),1.08(dd,J=37.8,5.8Hz,6H); ESI-MS:m / z=362.16[M+H] + .

[0062]

[0063] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced with 3-(2-((2R,6R)-2,6-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one (124.7 mg, 0.3454 mmol, 1.0 eq) to obtain 33.5 mg of a yellow-green solid (compound I-6), with a yield of 24.8%. 1H NMR(400MHz,DMSO-d6)δ11.81(s,1H),10.38(s,1H),7.40(s,1H),3.96(s,1H),3.88(s,3H),3.68(d,J =8.2Hz,1H),3.52-3.12(m,6H),2.31(s,3H),1.09(dd,J=38.2,6.2Hz,6H); ESI-MS:m / z=412.13[M+Na] + .

[0064] Example 7: Preparation of 3-(2-((3R,5S)-3,5-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromene-8-carbaldehyde (Compound I-7)

[0065] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (3R,5S)-3,5-dimethylmorpholine (129.3 mg, 1.1240 mmol, 1.2 eq) to obtain 245.7 mg of an off-white solid (3-(2-((3R,5S)-3,5-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one) in a yield of 72.6%. 1 HNMR (400MHz, DMSO-d6) δ10.18(s,1H),7.14(s,1H),6.76(s,1H),4.10(d,J=35.6Hz,2H),3. 83(s,3H),3.72-3.53(m,6H),2.28(s,3H),1.33(d,J=41.6Hz,6H); ESI-MS:m / z=362.16[M+H] + .

[0066]

[0067] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced with 3-(2-((3R,5S)-3,5-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one (124.7 mg, 0.3454 mmol, 1.0 eq) to obtain 36.2 mg of a yellow-green solid (compound I-7), with a yield of 26.8%. 1H NMR (400MHz, DMSO-d6) δ11.83(s,1H),10.40(s,1H),7.41(s,1H),4.14(d,J=41.2Hz,2H),3.91(s,3 H),3.64(dd,J=90.6,24.8Hz,6H),2.33(s,3H),1.31(d,J=103.8Hz,6H); ESI-MS:m / z=412.13[M+Na] + .

[0068] Example 8: Preparation of 3-(2-((2R,6S)-2,6-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromene-8-carbaldehyde (Compound I-8)

[0069] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (2R,6S)-2,6-dimethylmorpholine (129.3 mg, 1.1240 mmol, 1.2 eq) to obtain 224.4 mg of an off-white solid (3-(2-((2R,6S)-2,6-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one) in a yield of 66.4%. 1 H NMR (400MHz, DMSO-d6) δ10.15(s,1H),7.14(s,1H),6.74(s,1H),4.16(d,J=13.0Hz,1H),3.95(d,J=13.2Hz ,1H),3.84(s,3H),3.76-3.51(m,6H),2.28(s,3H),1.08(dd,J=18.8,5.4Hz,6H); ESI-MS:m / z=362.16[M+H] + .

[0070]

[0071] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced with 3-(2-((2R,6S)-2,6-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one (124.7 mg, 0.3454 mmol, 1.0 eq) to obtain 31.8 mg of a yellow-green solid (compound I-8), with a yield of 25.6%.1 H NMR (400MHz, DMSO-d6) δ11.78(s,1H),10.38(s,1H),7.40(s,1H),4.17(d,J=12.8Hz,1H),3.95(d,J=12.8Hz ,1H),3.88(s,3H),3.80-3.26(m,6H),2.31(s,3H),1.09(dd,J=21.6,6.2Hz,6H); ESI-MS:m / z=412.13[M+Na] + .

[0072] Example 9: Preparation of 3-(2-((3R,5R)-3,5-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromene-8-carbaldehyde (Compound I-9)

[0073] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (3R,5R)-3,5-dimethylmorpholine hydrochloride (170.4 mg, 1.1240 mmol, 1.2 eq) to obtain 235.4 mg of an off-white solid (3-(2-((3R,5R)-3,5-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one) in a yield of 69.6%. 1 HNMR(400MHz,DMSO-d6)δ10.15(s,1H),7.14(s,1H),6.74(s,1H),3.96(d,J=10.2Hz,2H),3. 84(s,3H),3.74-3.39(m,6H),2.28(s,3H),1.26(d,J=24.6Hz,6H); ESI-MS:m / z=362.16[M+H] + .

[0074]

[0075] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced with 3-(2-((3R,5R)-3,5-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one (124.7 mg, 0.3454 mmol, 1.0 eq) to obtain 39.8 mg of a yellow-green solid (compound I-9), with a yield of 29.5%.1 H NMR (400 MHz, DMSO-d6) δ 11.77 (s, 1H), 10.38 (s, 1H), 7.40 (s, 1H), 4.08-3.90 (m, 4H), 3.88 (s, 3H), 3.77-3.50 (m, 4H), 2.31 (s, 3H), 1.30 (s, 6H); ESI-MS: m / z = 412.13 [M+Na] + .

[0076] Example 10: Preparation of 3-(2-((3S,5S)-3,5-dimethylmorpholino)-2- oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-8-carboxaldehyde (Compound I-10)

[0077] This example employed the same procedure as Example 1, except that (R)-3- methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) was replaced by (3S,5S)-dimethylmorpholine (129.3 mg, 1.1240 mmol, 1.2 eq) in Step 3 to give a white solid (3-(2-((3S,5S)-3,5-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4- methyl-2H-chromen-2-one) 250.7 mg, yield: 74%. 1 H NMR (400 MHz, DMSO-d6) δ 10.15 (s, 1H), 7.14 (s, 1H), 6.75 (s, 1H), 3.96 (d, J = 9.4 Hz, 2H), 3.84 (s, 3H), 3.72-3.51 (m, 6H), 2.29 (s, 3H), 1.26 (d, J = 20.0 Hz, 6H); ESI-MS: m / z = 362.16 [M+H] + .

[0078]

[0079] This example employed the same procedure as Example 1, except that (R)-3- methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) was replaced by (3S,5S)-dimethylmorpholine (129.3 mg, 1.1240 mmol, 1.2 eq) in Step 3 to give a white solid (3-(2-((3S,5S)-3,5-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4- methyl-2H-chromen-2-one) 250.7 mg, yield: 74%. 1H NMR (400 MHz, DMSO-d6) δ 11.78 (s, 1H), 10.38 (s, 1H), 7.40 (s, 1H), 4.09-3.91 (m, 4H), 3.88 (s, 3H), 3.79-3.49 (m, 4H), 2.31 (s, 3H), 1.30 (s, 6H); ESI-MS: m / z = 412.13 [M+Na] + .

[0080] Example 11: Preparation of 3-(2-(1,4-oxazepan-4-yl)-2-oxoethyl)-7-hydroxy-6- methoxy-4-methyl-2-oxo-2H-chromen-8-carboxaldehyde (Compound I-11)

[0081] This example employed the same procedure as Example 1, except that (R)-3- methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) was replaced by 1,4-oxazepane (113.5 mg, 1.1240 mmol, 1.2 eq) in Step 3 to give a white solid (3-(2-(1,4-oxazepan-4-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2- one) 213.2 mg, yield: 65.6%. 1 H NMR (400 MHz, DMSO-d6) δ 10.15 (s, 1H), 7.14 (s, 1H), 6.74 (s, 1H), 3.84 (s, 3H), 3.75-3.50 (m, 10H), 2.29 (s, 3H), 1.91-1.68 (m, 2H); ESI-MS: m / z = 348.14 [M+H] + .

[0082]

[0083] This example employed the same procedure as Example 1, except that (R)-3- methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) was replaced by 1,4-oxazepane (113.5 mg, 1.1240 mmol, 1.2 eq) in Step 3 to give a white solid (3-(2-(1,4-oxazepan-4-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2- one) 213.2 mg, yield: 65.6%. 1HNMR (400 MHz, DMSO-d6) δ 11.78 (s, 1H), 10.38 (s, 1H), 7.41 (s, 1H), 3.88 (s, 3H), 3.76-3.51 (m, 10H), 2.31 (s, 3H), 2.00-1.85 (m, 1H), 1.76-1.63 (m, 1H); ESI-MS: m / z = 398.12 [M+Na] + .

[0084] Example 12: Preparation of 2-(8-formyl-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)-N-((1r,4r)-4-methoxycyclohexyl)acetamide (Compound I-12)

[0085] This example employed the same procedure as Example 1 except that (R)-3- methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) was replaced by (1r,4r)-4- methoxycyclohexan-1-amine (145.0 mg, 1.1240 mmol, 1.2 eq) in Step 3 to give a white solid (2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)-N-((1r,4r)-4- methoxycyclohexyl)acetamide) 196 mg, yield: 56.0%. 1 HNMR (400 MHz, DMSO-d6) δ 11.78 (s, 1H), 10.38 (s, 1H), 7.41 (s, 1H), 3.88 (s, 3H), 3.76-3.51 (m, 10H), 2.31 (s, 3H), 2.00-1.85 (m, 1H), 1.76-1.63 (m, 1H); ESI-MS: m / z = 398.12 [M+Na] + .

[0086]

[0087] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced by 2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)-N-((1r,4r)-4-methoxycyclohexyl)acetamide (129.6 mg, 0.3454 mmol, 1.0 eq) to obtain 17.4 mg of a yellow-green solid (compound I-12), with a yield of 12.4%. 1 HNMR (400MHz, DMSO-d6) δ11.76(s,1H),10.37(s,1H),7.83(d,J=7.8Hz,1H),7.38(s,1H),3.87(s,3H),3.53-3.44(m,1H),3.42(s,2H),3. 18(s,3H),3.10-3.00(m,1H),2.32(s,3H),1.93(d,J=11.0Hz,2H),1.74(d,J=11.2Hz,2H),1.23-1.06(m,4H); ESI-MS:m / z=426.15[M+Na] + .

[0088] Example 13: Preparation of 3-(2-(3,3-bis(hydroxymethyl)azetidin-1-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromene-8-carbaldehyde (Compound I-13)

[0089] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with 2-oxa-6-azaspiro[3.3]heptane (111.3 mg, 1.1240 mmol, 1.2 eq) to obtain 207.6 mg of an off-white solid (7-hydroxy-6-methoxy-4-methyl-3-(2-oxo-2-(2-oxa-6-azaspiro[3.3]hept-6-yl)ethyl)-2H-chromen-2-one) in a yield of 64.2%. 1 H NMR (400MHz, DMSO-d6) δ10.16(s,1H),7.13(s,1H),6.73(s,1H),4.65(d,J=5.4Hz,2H),4.37(s, 2H),3.99(s,2H),3.83(s,3H),3.34(s,2H),2.81(s,2H),2.30(s,3H); ESI-MS: m / z=346.12[M+H] + .

[0090]

[0091]

[0092] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced with 7-hydroxy-6-methoxy-4-methyl-3-(2-oxo-2-(2-oxa-6-azaspiro[3.3]hept-6-yl)ethyl)-2H-chromen-2-one (119.2 mg, 0.3454 mmol, 1.0 eq) to obtain 20.3 mg of a yellow-green solid (compound I-13), with a yield of 15.0%. 1 H NMR(400MHz,DMSO-d6)δ11.77(s,1H),10.37(s,1H),7.39(s,1H),4.86(s,2H),3.91(s,2H), 3.88(s,3H),3.52(s,2H),3.48(s,4H),3.39(s,2H),2.33(s,3H); ESI-MS: m / z=414.12[M+Na] + .

[0093] Example 14: Preparation of 3-(2-(4,4-bis(hydroxymethyl)piperidin-1-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromene-8-carbaldehyde (Compound I-14)

[0094] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with 2-oxa-7-azaspiro[3.5]nonane (142.7 mg, 1.1240 mmol, 1.2 eq) to obtain 221.4 mg of an off-white solid (7-hydroxy-6-methoxy-4-methyl-3-(2-oxo-2-(2-oxa-7-azaspiro[3.5]non-7-yl)ethyl)-2H-chromen-2-one) in a yield of 63.4%. 1HNMR(400MHz,DMSO-d6)δ10.15(s,1H),7.13(s,1H),6.74(s,1H),4.31(s,4H),3.83(s,3H),3.61 (s,2H),3.39(d,J=48.8Hz,5H),2.26(s,3H),1.82(s,2H),1.67(s,2H); ESI-MS:m / z=374.16[M+H] + .

[0095]

[0096] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced with 7-hydroxy-6-methoxy-4-methyl-3-(2-oxo-2-(2-oxa-7-azaspiro[3.5]non-7-yl)ethyl)-2H-chromen-2-one (128.8 mg, 0.3454 mmol, 1.0 eq) to give 17.1 mg of a yellow-green solid (compound I-14) with a yield of 11.9%. 1 HNMR(400MHz,DMSO-d6)δ11.79(s,1H),10.38(s,1H),7.39(s,1H),4.43(s,2H),3.88(s,3H),3.62( s,2H),3.53-3.36(m,4H),3.31(s,4H),2.29(s,3H),1.47-1.24(m,4H); ESI-MS:m / z=442.15[M+Na] + .

[0097] Example 15: Preparation of 3-(2-(8-oxa-3-azabicyclo[3.2.1]oct-3-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-benzopyran-8-carbaldehyde (Compound I-15)

[0098] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with 8-oxo-3-azabicyclo[3,2,1]octane hydrochloride (168.2 mg, 1.1240 mmol, 1.2 eq) to obtain 224.7 mg of an off-white solid (3-(2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one) in a yield of 66.8%.1 H NMR(400MHz,DMSO-d6)δ10.17(s,1H),7.14(s,1H),6.75(s,1H),4.33-4.27(m,2H),3.8 3(s,3H),3.77-3.54(m,6H),2.28(s,3H),1.21-1.21(m,4H); ESI-MS:m / z=360.14[M+H] + .

[0099]

[0100] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced with 3-(2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one (124.0 mg, 0.3454 mmol, 1.0 eq) to obtain 26.2 mg of a yellow-green solid (compound I-15), with a yield of 19.5%. 1 HNMR (400MHz, DMSO-d6) δ11.80(s,1H),10.39(s,1H),7.41(s,1H),4.30(d,J=6.0Hz,2H),3.89(s,3H),3.76(t,J=13.4Hz,2H),3.48(d,J =16.6Hz,1H),3.33(d,J=12.8Hz,2H),2.82-2.73(m,1H),2.31(s,3H),1.90-1.70(m,3H),1.59-1.49(m,1H); ESI-MS: m / z=410.12[M+Na] + .

[0101] Example 16: Preparation of 7-hydroxy-6-methoxy-4-methyl-2-oxo-3-(2-oxo-2-((3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)ethyl)-2H-chromene-8-carbaldehyde (Compound I-16)

[0102] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (3aR,6aS)-hexahydro-1H-furo[3,4-c]pyrrole hydrochloride (168.2 mg, 1.1240 mmol, 1.2 eq) to obtain 227.8 mg of an off-white solid (7-hydroxy-6-methoxy-4-methyl-3-(2-oxo-2-((3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)ethyl)-2H-chromen-2-one) in a yield of 67.7%. 1 H NMR (400MHz, DMSO-d6) δ10.15(s,1H),7.13(s,1H),6.74(s,1H),3.83(s,3H),3. 81-3.45(m,10H),2.92(d,J=44.8Hz,2H),2.29(s,3H); ESI-MS:m / z=360.14[M+H] + .

[0103]

[0104] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced with 7-hydroxy-6-methoxy-4-methyl-3-(2-oxo-2-((3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)ethyl)-2H-chromen-2-one (124.0 mg, 0.3454 mmol, 1.0 eq) to give 33.1 mg of a yellow-green solid (compound I-16), with a yield of 24.6%. 1 HNMR(400MHz,DMSO-d6)δ11.79(s,1H),10.39(s,1H),7.42(s,1H),3.89(s,3H),3.85-3.4 7(m,10H),3.03-2.94(m,1H),2.90-2.82(m,1H),2.33(s,3H); ESI-MS:m / z=410.12[M+Na] + .

[0105] Example 17: Preparation of 3-(2-((1R,4R)-2-oxo-5-azabicyclo[2.2.1]heptane-5-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-benzopyran-8-carbaldehyde (Compound I-17)

[0106] This example employs the same implementation as Example 1, except that (R)-3- methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (1R,4R)-2-oxa-5- azabicyclo[2.2.1]heptane hydrochloride (152.4 mg, 1.1240 mmol, 1.2 eq) in Step 3 to give a white solid (3-(2-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptan-5-yl)-2-oxoethyl)-7- hydroxy-6-methoxy-4-methyl-2H-chromen-2-one) 183.5 mg, yield: 56.9%. 1 H NMR (400 MHz, DMSO-d6) δ 10.17 (s, 1H), 7.14 (s, 1H), 6.74 (s, 1H), 4.61 (d, J = 39.4 Hz, 2H), 3.83 (s, 3H), 3.76 - 3.44 (m, 6H), 2.32 (s, 3H), 1.89 - 1.71 (m, 2H); ESI-MS: m / z = 346.12 [M+H] + .

[0107]

[0108] This example employs the same implementation as Example 1, except that (R)-3- methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (1R,4R)-2-oxa-5- azabicyclo[2.2.1]heptane hydrochloride (152.4 mg, 1.1240 mmol, 1.2 eq) in Step 3 to give a white solid (3-(2-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptan-5-yl)-2-oxoethyl)-7- hydroxy-6-methoxy-4-methyl-2H-chromen-2-one) 183.5 mg, yield: 56.9%. 1 H NMR (400 MHz, DMSO-d6) δ 10.17 (s, 1H), 7.14 (s, 1H), 6.74 (s, 1H), 4.61 (d, J = 39.4 Hz, 2H), 3.83 (s, 3H), 3.76 - 3.44 (m, 6H), 2.32 (s, 3H), 1.89 - 1.71 (m, 2H); ESI-MS: m / z = 346.12 [M+H] + .

[0109] Example 18: Preparation of 3-(2-((1S,4S)-2-oxo-5-azabicyclo[2.2.1]heptane-5-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-benzopyran-8-carbaldehyde (Compound I-18)

[0110] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptane hydrochloride (152.4 mg, 1.1240 mmol, 1.2 eq) to obtain 201.6 mg of an off-white solid (3-(2-((1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one) in a yield of 63.3%. 1 H NMR (400MHz, DMSO-d6) δ10.16(s,1H),7.14(s,1H),6.74(s,1H),4.61(d,J=38.4Hz,2H),3.8 4(s,3H),3.71-3.46(m,6H),2.32(s,3H),1.79(d,J=38.4Hz,2H); ESI-MS:m / z=346.12[M+H] + .

[0111]

[0112] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (119.2 mg, 0.3454 mmol, 1.0 eq) is replaced by 3-(2-((1S,4S)-2-oxa-5-azabicyclo[2.2.1]hept-5-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) to obtain 25.2 mg of a yellow-green solid (compound I-18), with a yield of 19.5%. 1HNMR(400MHz,DMSO-d6)δ11.78(s,1H),10.37(s,1H),7.39(s,1H),4.84-4.56(m,2H),3.88(s,3H),3.83-3.66(m,2H), 3.62-3.41(m,3H),3.19(dd,J=26.2,11.6Hz,1H),2.33(d,J=5.8Hz,3H),1.92-1.71(m,2H); ESI-MS:m / z=396.11[M+Na] + .

[0113] Example 19: Preparation of 7-hydroxy-6-methoxy-4-methyl-2-oxo-3-(2-oxo-2-(2-oxa-8-azaspiro[4.5]dec-8-yl)ethyl)-2H-chromene-8-carbaldehyde (Compound I-19)

[0114] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with 2-oxa-8-azaspiro[4.5]decane hydrochloride (199.6 mg, 1.1240 mmol, 1.2 eq) to obtain 235.0 mg of an off-white solid (7-hydroxy-6-methoxy-4-methyl-3-(2-oxo-2-(2-oxa-8-azaspiro[4.5]decan-8-yl)ethyl)-2H-chromen-2-one) in a yield of 64.8%. 1 H NMR (400MHz, DMSO-d6) δ10.15(s,1H),7.14(s,1H),6.75(s,1H),3.86(s,3H),3.73(dd,J=12.8,6.8Hz,2H),3.63-3.45(m ,8H),2.28(s,3H),1.72(dd,J=13.2,6.6Hz,2H),1.54(d,J=4.6Hz,2H),1.40(d,J=4.2Hz,2H); ESI-MS: m / z=388.17[M+H] + .

[0115]

[0116] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced by 7-hydroxy-6-methoxy-4-methyl-3-(2-oxo-2-(2-oxa-8-azaspiro[4.5]decan-8-yl)ethyl)-2H-chromen-2-one (133.7 mg, 0.3454 mmol, 1.0 eq) to obtain 34.3 mg of a yellow-green solid (compound I-19), with a yield of 23.8%. 1 H NMR (400MHz, DMSO-d6) δ11.82(s,1H),10.41(s,1H),7.43(s,1H),3.91(s,3H),3.76(t,J=7.2Hz,2H),3.70-3.49(m, 6H),3.39-3.31(m,2H),2.33(s,3H),1.75(t,J=7.2Hz,2H),1.51(dt,J=61.2,5.4Hz,4H); ESI-MS:m / z=438.15[M+Na] + .

[0117] Example 20: Preparation of 7-hydroxy-6-methoxy-4-methyl-2-oxo-3-(2-oxo-2-(8-oxa-2-azaspiro[4.5]dec-2-yl)ethyl)-2H-chloromethane-8-carbaldehyde (Compound I-20)

[0118] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with 8-oxa-2-azaspiro[4.5]decane hydrochloride (199.6 mg, 1.1240 mmol, 1.2 eq) to obtain 215.1 mg of an off-white solid (7-hydroxy-6-methoxy-4-methyl-3-(2-oxo-2-(8-oxa-2-azaspiro[4.5]dec-2-yl)ethyl)-2H-chromen-2-one) in a yield of 59.3%. 1 H NMR (400MHz, DMSO-d6) δ10.14(s,1H),7.12(s,1H),6.73(s,1H),3.83(s,3H),3.71-3.39(m,10H),2.28(s, 3H), 1.83 (t, J=6.8Hz, 1H), 1.71 (t, J=6.8Hz, 1H), 1.47 (dd, J=11.8, 4.8Hz, 4H); ESI-MS: m / z=388.17[M+H] + .

[0119]

[0120] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced with 7-hydroxy-6-methoxy-4-methyl-3-(2-oxo-2-(8-oxa-2-azaspiro[4.5]dec-2-yl)ethyl)-2H-chromen-2-one (133.7 mg, 0.3454 mmol, 1.0 eq) to obtain 41.5 mg of a yellow-green solid (compound I-20), with a yield of 28.8%. 1 HNMR (400MHz, DMSO-d6) δ11.78(s,1H),10.38(s,1H),7.40(s,1H),3.88(s,3H),3.72-3.45(m,8H),3.32(dd,J=13. 6,6.4Hz,2H),2.32(s,3H),1.78(dt,J=48.8,7.2Hz,2H),1.49(dt,J=18.8,5.6Hz,4H); ESI-MS:m / z=438.15[M+Na] + .

[0121] Example 21: Preparation of N-(1,3-dihydroisobenzofuran-5-yl)-2-(8-formyl-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamide (Compound I-21)

[0122] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with 1,3-dihydroisobenzofuran-5-amine (151.9 mg, 1.1240 mmol, 1.2 eq). The reaction time is 5 h to obtain 175.9 mg of an off-white solid (N-(1,3-dihydroisobenzofuran-5-yl)-2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamide) in a yield of 49.2%. 1 H NMR (400MHz, DMSO-d6) δ10.20(s,1H),10.12(s,1H),7.56(s,1H),7.39(d,J=8.2Hz,1H),7.18(d,J=11 .4Hz,2H),6.75(s,1H),4.92(s,4H),3.84(s,3H),3.67(s,2H),2.38(s,3H); ESI-MS:m / z=382.12[MH]+ .

[0123]

[0124] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced with N-(1,3-dihydroisobenzofuran-5-yl)-2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamide (131.6 mg, 0.3454 mmol, 1.0 eq), the amount of HMTA used is 193.6 mg (1.3819 mmol, 4.0 eq), the reaction time is 3 h, the reaction temperature is 120 ° C, and 12.4 mg of brown solid (compound I-21) is obtained with a yield of 8.7%. 1 HNMR(400MHz,DMSO-d6)δ11.79(s,1H),10.39(s,1H),10.15(s,1H),7.56(s,1H),7.42(s,1H),7.39(d,J=7.0H z,1H),7.18(d,J=8.2Hz,1H),4.91(s,4H),3.89(s,3H),3.71(s,2H),2.41(s,3H); ESI-MS:m / z=432.11[M+Na] + .

[0125] Example 22: Preparation of 2-(8-formyl-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)-N-(5-methoxypyridin-2-yl)acetamide (Compound I-22)

[0126] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with 2-amino-5-methoxypyridine (139.4 mg, 1.1240 mmol, 1.2 eq). The reaction time is 5 h to obtain 190.7 mg of an off-white solid (2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)-N-(5-methoxypyridin-2-yl)acetamide) with a yield of 54.9%. 1H NMR(400MHz,DMSO-d6)δ10.48(s,1H),10.18(s,1H),8.01(s,1H),7.92(s,1H),7.38(s,1H),7.16( s,1H),6.75(s,1H),3.84(s,3H),3.76(s,3H),3.71(s,2H),2.36(s,3H); ESI-MS:m / z=371.12[M+H] + .

[0127]

[0128] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced by 2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)-N-(5-methoxypyridin-2-yl)acetamide (127.8 mg, 0.3454 mmol, 1.0 eq) to obtain 27.1 mg of dark green solid (compound I-22), yield: 20.8%. 1 H NMR (400MHz, DMSO-d6) δ11.80(s,1H),10.51(s,1H),10.38(s,1H),8.00(d,J=2.2Hz,1H),7.91(d,J=9.0Hz,1H ),7.41(s,1H),7.36(dd,J=9.2,2.6Hz,1H),3.89(s,3H),3.76(s,5H),2.39(s,3H); ESI-MS:m / z=399.12[M+H] + .

[0129] Example 23: Preparation of 2-(8-formyl-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)-N-(6-methoxypyridin-3-yl)acetamide (Compound I-23)

[0130] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with 5-amino-2-methoxypyridine (139.4 mg, 1.1240 mmol, 1.2 eq). The reaction time is 5 h to obtain 204.2 mg of an off-white solid (2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)-N-(6-methoxypyridin-3-yl)acetamide) with a yield of 58.8%. 1H NMR(400MHz,DMSO-d6)δ10.10(s,2H),8.32(s,1H),7.85(s,1H),7.17(s,1H),6.76(s ,2H),3.84(s,3H),3.77(s,3H),3.66(s,2H),2.38(s,3H); ESI-MS: m / z=371.12[M+H] + .

[0131]

[0132] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced by 2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)-N-(6-methoxypyridin-3-yl)acetamide (127.8 mg, 0.3454 mmol, 1.0 eq) to obtain 23.7 mg of dark green solid (compound I-23), yield: 18.2%. 1 H NMR (400MHz, DMSO-d6) δ11.77(s,1H),10.39(s,1H),10.12(s,1H),8.32(d,J=2.4Hz,1H),7.84(dd,J=8.8,2.2Hz,1H ),7.42(s,1H),6.75(d,J=8.8Hz,1H),3.89(s,3H),3.77(s,3H),3.70(s,2H),2.41(s,3H); ESI-MS: m / z=399.12[M+H] + .

[0133] Example 24: Preparation of methyl 5-(2-(8-formyl-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamido)nicotinate (Compound I-24)

[0134] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with 5-aminopyridine-3-carboxylic acid methyl ester (170.8 mg, 1.1240 mmol, 1.2 eq). The reaction time is 5 h to obtain 238.3 mg of an off-white solid (5-(2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamido)nicotinate) in a yield of 64.0%. 1HNMR(400MHz,DMSO-d6)δ10.56(s,1H),10.22(s,1H),8.90(s,1H),8.73(s,1H),8.59(s,1H) ,7.18(s,1H),6.77(s,1H),3.85(s,6H),3.73(s,3H),2.40(s,3H); ESI-MS:m / z=399.11[M+H] + .

[0135]

[0136] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced with 5-(2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamido)nicotinate methyl ester (137.8 mg, 0.3454 mmol, 1.0 eq) to obtain 38.5 mg of a yellow-green solid (compound I-24), with a yield of 27.5%. 1 H NMR(400MHz,DMSO-d6)δ11.82(s,1H),10.59(s,1H),10.41(s,1H),8.90(s,1H),8.73(s,1H),8.58( s,1H),7.46(s,1H),3.91(s,3H),3.84(s,3H),3.78(s,2H),2.44(s,3H); ESI-MS:m / z=427.11[M+H] + .

[0137] Example 25: Preparation of 5-(2-(8-formyl-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetylamino)nicotinic acid (Compound I-25)

[0138]

[0139] This example follows the same protocol as Example 24, except that Step 5 is added: 5-(2-(8-formyl-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamido)nicotinate (50.0 mg, 0.1238 mmol) was added all at once to 2 mL of 4 mol / L NaOH solution and stirred at room temperature for 2 h. The reaction mixture was acidified to pH 3 with 1 mol / L HCl. The resulting precipitate was filtered, eluted with ethyl acetate, and dried under vacuum to afford 11.5 mg of a brown solid (Compound I-25) in a 23.8% yield.1 H NMR(400MHz,DMSO-d6)δ13.41(s,1H),10.53(s,1H),9.42(s,2H),8.90(s,1H),8.72(s,1H), 8.55(s,1H),6.78(s,1H),3.84(s,3H),3.74(s,2H),2.38(s,3H); ESI-MS: m / z=413.10[M+H] + .

[0140] Example 26: Preparation of N-(5-chloropyridin-3-yl)-2-(8-formyl-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamide (Compound I-26)

[0141] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with 3-amino-5-chloropyridine (144.5 mg, 1.1240 mmol, 1.2 eq). The reaction time is 5 h to obtain 217.5 mg of an off-white solid (N-(5-chloropyridin-3-yl)-2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamide) in a yield of 62.1%. 1 HNMR(400MHz,DMSO-d6)δ10.56(s,1H),10.22(s,1H),8.61(s,1H),8.28(s,1H),8.19(s,1H) ,7.17(s,1H),6.76(s,1H),3.85(s,3H),3.72(s,2H),2.39(s,3H); ESI-MS: m / z=375.06[M+H] + .

[0142]

[0143] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced by N-(5-chloropyridin-3-yl)-2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamide (128.7 mg, 0.3454 mmol, 1.0 eq) to obtain 37.4 mg of dark green solid compound I-26) with a yield of 28.6%. 1HNMR(400MHz,DMSO-d6)δ11.81(s,1H),10.59(s,1H),10.40(s,1H),8.61(s,1H),8.28(s,1H) ,8.18(s,1H),7.44(s,1H),3.90(s,3H),3.77(s,2H),2.43(s,3H).; ESI-MS: m / z=403.07[M+H] + .

[0144] Example 27: Preparation of N-(6-chloropyridin-3-yl)-2-(8-formyl-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamide (Compound I-27)

[0145] This example adopts the same implementation method as Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with 2-chloro-5-aminopyridine (144.5 mg, 1.1240 mmol, 1.2 eq). The reaction time is 5 h to obtain 195.3 mg of an off-white solid (N-(6-chloropyridin-3-yl)-2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamide) in a yield of 55.6%. 1 HNMR(400MHz,DMSO-d6)δ10.47(s,1H),10.21(s,1H),8.57(s,1H),8.02(s,1H),7.42(s,1H) ,7.17(s,1H),6.76(s,1H),3.84(s,3H),3.70(s,2H),2.38(s,3H); ESI-MS:m / z=375.06[M+H] + .

[0146]

[0147] The difference is that in step 4, (R)-7-hydroxy-6-methoxy-4-methyl-3-(2-(3-methylmorpholino)-2-oxoethyl)-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced by N-(6-chloropyridin-3-yl)-2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamide (128.7 mg, 0.3454 mmol, 1.0 eq) to obtain 29.3 mg of dark green solid (compound I-27), yield: 22.2%. 1HNMR (400 MHz, DMSO-d6) δ 11.82 (s, 1H), 10.50 (s, 1H), 10.40 (s, 1H), 8.58 (s, 1H), 8.02 (d, J = 8.8 Hz, 1H), 7.43 (d, J = 10.0 Hz, 2H), 3.90 (s, 3H), 3.75 (s, 2H), 2.42 (s, 3H); ESI-MS: m / z = 403.07 [M+H] + .

[0148] Example 28: Preparation of N-(5-chloropyridin-2-yl)-2-(8-formyl-7-hydroxy-6- methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamide (Compound I-28)

[0149] This example employed the same procedure as in Example 1 except that (R)-3- methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) was replaced by 2-amino-5- chloropyridine (144.5 mg, 1.1240 mmol, 1.2 eq) in Step 3 and the reaction time was 5 h to give a white solid (N-(5-chloropyridin-2-yl)-2-(7-hydroxy-6-methoxy-4-methyl-2- oxo-2H-chromen-3-yl)acetamide) 207.4 mg in 59.2% yield. 1 HNMR (400 MHz, DMSO-d6) δ 10.83 (s, 1H), 10.20 (s, 1H), 8.35 (s, 1H), 8.02 (d, J = 8.8 Hz, 1H), 7.85 (d, J = 8.8 Hz, 1H), 7.14 (d, J = 12.2 Hz, 1H), 6.76 (s, 1H), 3.84 (s, 3H), 3.76 (s, 2H), 2.36 (s, 3H); ESI-MS: m / z = 375.06 [M+H] + .

[0150]

[0151] This example employed the same procedure as in Example 1 except that (R)-3- methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) was replaced by 2-amino-5- chloropyridine (144.5 mg, 1.1240 mmol, 1.2 eq) in Step 3 and the reaction time was 5 h to give a white solid (N-(5-chloropyridin-2-yl)-2-(7-hydroxy-6-methoxy-4-methyl-2- oxo-2H-chromen-3-yl)acetamide) 207.4 mg in 59.2% yield. 1HNMR(400MHz,DMSO-d6)δ11.82(s,1H),10.86(s,1H),10.41(s,1H),8.35(s,1H),8.01(d,J=8.4Hz,1H) ,7.85(d,J=8.8Hz,1H),7.46(s,1H),3.90(s,3H),3.81(s,2H),2.41(s,3H); ESI-MS:m / z=403.07[M+H] + .

[0152] Example 29: Proliferation inhibition activity of the compounds of the present invention on mouse lung fibroblast cell line Mlg2908 and human non-small cell lung cancer cell line H358

[0153] The inhibitory activity of the compounds of the present invention on Mlg2908 and H358 cells was determined using the Cell Counting Kit-8. Three replicate wells were set up for each concentration of the test compound. A blank control group, in which only culture medium was added but no cells were present, and a negative control group, in which cells were inoculated but no drug was administered, were set up for each group. Three replicate wells were set up for each group. Cells in the logarithmic growth phase were digested, perfused into a single-cell suspension, and seeded into 96-well culture plates. Both Mlg2908 and H358 cells were plated at 5×10 3 Cells / well. Add 100 μL of complete culture medium to each well and culture in a 5% carbon dioxide incubator for 24 hours. Discard the culture medium, add 200 μL of drug-containing culture medium to each well and culture for another 72 hours. The drug concentration is diluted threefold starting from 30 μM to set 9 concentrations. After 72 hours, add 20 μL of Cell Counting Kit-8 reagent to each well and incubate in the incubator for 3 hours. The OD value of each well is detected by enzyme reader (detection wavelength: 450 nm), and the inhibition rate is calculated according to the following formula: Inhibition rate (%) = (OD negative - OD administration) / (OD negative - OD blank) × 100%, IC 50 The values ​​were obtained by processing with Graphpad 5.0 software.

[0154] Table 1 Inhibitory activity of the compounds of the present invention on proliferation of Mlg2908 cells and H358 cells

[0155]

[0156]

[0157] From the proliferation inhibitory activity data on Mlg2908 cells and H358 cells in Table 1, it can be seen that most compounds exhibit good proliferation inhibitory activity on Mlg2908 cells and H358 cells, and some compounds have better cell proliferation inhibitory activity than Orin1001, indicating good application prospects.

Claims

1. A polysubstituted coumarin compound, the structural formula of which is as follows: （I） in, R1 is selected from 、 、 、 、 、 、 、 、 、 、 、 、 、 、 or .

2. A polysubstituted coumarin compound according to claim 1 in free form or a pharmaceutically acceptable salt thereof.

3. Use of a polysubstituted coumarin compound according to claim 1 in the preparation of a drug for treating diseases mediated by inositol-requiring enzyme 1α (IRE1α).

4. The use according to claim 3, characterized in that The disease mediated by the inositol-requiring enzyme 1α (IRE1α) is cancer or pulmonary fibrosis; the cancer is lung cancer, bronchial cancer, prostate cancer, breast cancer, pancreatic cancer, colon cancer, rectal cancer, thyroid cancer, liver cancer, gastric cancer, glioma, glioblastoma, endometrial cancer, kidney cancer, bladder cancer, uterine body cancer, cervical cancer, ovarian cancer, multiple myeloma, esophageal cancer, lymphocytic leukemia, myeloid leukemia, brain cancer, oral cancer, small intestine cancer, non-Hodgkin's lymphoma, and melanoma.

5. The use according to claim 4, wherein the cancer is villous colon adenoma, intrahepatic bile duct carcinoma, hepatocellular carcinoma, renal pelvis carcinoma, acute myeloid leukemia, chronic myeloid leukemia, pharyngeal cancer, and laryngeal cancer.

6. A pharmaceutical composition containing the polysubstituted coumarin compound according to claim 1, characterized in that: The pharmaceutical composition consists of the polysubstituted coumarin compound and a pharmaceutically acceptable excipient.

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