Polysubstituted coumarin compound and application

Abstract

The invention provides a polysubstituted coumarin compound with a structure as shown in a general formula (I) as well as pharmaceutically acceptable salts, solvates and application thereof, wherein R1 is defined in the specification. The invention also relates to preparation of the compound, a pharmaceutical composition of the compound and application of the compound to treatment of related diseases by inhibiting inositol-demanding enzyme 1 alpha (IRE1 alpha), such as treatment of diseases such as cancer and pulmonary fibrosis. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of medicine, and specifically relates to a polysubstituted coumarin compound and an application and a preparation method thereof. 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 response is regulated by the unfolded protein response (UPR), which is an integrated adaptive pathway 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 UPR is triggered, allowing cells to restore homeostasis and restore the endoplasmic reticulum to a normal physiological state. However, in the case of persistent or severe ERS, the UPR's ability to restore cells is broken, leading to cell death. 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 essential 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, and activates 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 activation of 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 an active form of XBP1s. XBP1s then translocates to the nucleus and promotes the transcription of ERAD system components, including 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, and the endoribonuclease activity of oligomeric IRE1α is reduced, and the degradation of mRNA and microRNA (miRNA) is promoted through the IRE1α-dependent decay process (Hollien J, Lin JH, Li H, et al. Journal of Cell Biology, 2009, 186 (3): 323-331). PERK, ATF6, and IRE1α work together to maintain cell homeostasis in the ERS environment.

[0004] ORIN1001 is an IRE1α inhibitor developed by Fosun Hongchuang (Suzhou) Pharmaceutical Technology Co., Ltd. It has a good therapeutic effect 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] In view of the above problems, the present invention provides a polysubstituted coumarin compound and its application, which has favorable pharmacological properties, can inhibit IRE1α, and has better proliferation inhibition activity on Mlg2908 cells and H358 cells than ORIN1001 in clinical phase II. Therefore, the compound of formula (I) is suitable for treating diseases dependent on IRE1α, especially 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] Further, the disease mediated by 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 poly-substituted 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 the compound of formula (III): CH3SO3H was added to a suspension of 4-methoxyresorcinol (1.0 eq) in diethyl acetyl succinate (1.2 eq). 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 removed by vacuum distillation to obtain a crude product, which was then separated and purified by a silica gel column to obtain the compound of formula (III).

[0024] Synthesis of the compound of formula (IV): The compound of formula (III) was added once to a 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 precipitate formed was filtered, filtered with ethyl acetate and dried in vacuo to obtain the compound of formula (IV).

[0025] Synthesis of the compound of formula (V): Dissolve the compound of formula (IV) (1.0 eq) and HATU (1.5 eq) in anhydrous DMF, and add DIPEA (4.0 eq) and the amine-containing compound R1 (1.2 eq) in sequence. Stir the mixture at room temperature for 3-5 h. Extract the organic layer with ethyl acetate, remove moisture with anhydrous sodium sulfate, and then separate and purify with a silica gel column to obtain the compound of formula (V).

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

[0027] The beneficial effects of the present invention are: having favorable pharmacological properties, being able to inhibit IRE1α, and having a proliferation inhibitory activity on Mlg2908 cells and H358 cells that is superior to ORIN1001 in clinical phase II. DETAILED DESCRIPTION

[0028] The specific embodiments included below are for illustration and should not be construed as limiting the scope of the invention. In addition, it should be understood that after reading the content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms fall within the scope limited by the appended claims of the application equally.

[0029] The synthetic route of polysubstituted coumarin compounds 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-chromene-8-carbaldehyde (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] CH3SO3H was added 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). 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 removed by vacuum distillation to obtain a crude product, which was then separated and purified by a silica gel column 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] 2-(7-Hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)ethyl acetate (60.0 g, 205 mmol) was added to 30 mL of 4 mol / L NaOH solution at once and stirred at room temperature for 4 h. The reaction mixture was acidified to pH = 3 with 5 mol / L HCl. The formed precipitate was filtered, filtered with ethyl acetate and dried under vacuum to obtain 39.4 g of a light 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] Dissolve 2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetic acid (240.0 mg, 0.9366 mmol, 1.0 eq) and HATU (534.2 mg, 1.4051 mmol, 1.5 eq) in anhydrous DMF, and add DIPEA (652.6 μL, 3.7466 mmol, 4.0 eq) and (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) in turn. Stir the mixture at room temperature for 3 h. Extract the organic layer with ethyl acetate, remove moisture with anhydrous sodium sulfate, and separate and purify with a silica gel column to obtain 242.2 mg of a white solid (Compound V-1). The yield is 74.5%. 1 H NMR (400MHz, DMSO-d6) δ10.16(s,1H),7.14(s,1H),6.75(s,1H),4.20(d,J=60.3Hz,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 with dichloromethane, and the water was removed by anhydrous sodium sulfate. Then, 40.0 mg of a yellow-green solid (Compound I-1) was obtained by separation and purification by a silica gel column. The yield was 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 that of 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) with 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 obtain 32.5 mg of a yellow-green solid (compound I-2), with 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 that of 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) with 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 obtain 35.7 mg of a yellow-green solid (compound I-3), with 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 that of 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) with 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 obtain 28.7 mg of a yellow-green solid (compound I-4), with 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 that of 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) with 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 by 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 that of 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) with 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 by 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 that of 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) with 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 by 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 that of 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) with 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 by 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 that of 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-chromene-2-one) with 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-chromene-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) is replaced by 3-(2-((3R,5R)-3,5-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromene-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(400MHz,DMSO-d6)δ11.77(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-chromene-8-carbaldehyde (Compound I-10)

[0077] This example adopts the same implementation method as that of Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (3S, 5S)-dimethylmorpholine (129.3 mg, 1.1240 mmol, 1.2 eq) to obtain 250.7 mg of an off-white solid (3-(2-((3S, 5S)-3,5-dimethylmorpholino)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one) with a yield of 74%. 1 H NMR (400MHz, DMSO-d6) δ10.15(s,1H),7.14(s,1H),6.75(s,1H),3.96(d,J=9.4Hz,2H),3.8 4(s,3H),3.72-3.51(m,6H),2.29(s,3H),1.26(d,J=20.0Hz,6H); ESI-MS:m / z=362.16[M+H] + .

[0078]

[0079] 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 3-(2-((3S,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 43.8 mg of a yellow-green solid (Compound I-10), with a yield of 32.4%. 1H NMR(400MHz,DMSO-d6)δ11.78(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-benzopyran-8-carbaldehyde (Compound I-11)

[0081] This example adopts the same implementation method as that of Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with 1,4-oxaazepine (113.5 mg, 1.1240 mmol, 1.2 eq) to obtain 213.2 mg of an off-white solid (3-(2-(1,4-oxazolidin-4-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one) with a yield of 65.6%. 1 H NMR(400MHz,DMSO-d6)δ10.15(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] 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 3-(2-(1,4-oxazolidin-4-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one (120.0 mg, 0.3454 mmol, 1.0 eq) to obtain 25.8 mg of a yellow-green solid (Compound I-11), with a yield of 19.8%. 1HNMR(400MHz,DMSO-d6)δ11.78(s,1H),10.38(s,1H),7.41(s,1H),3.88(s,3H),3.76-3.5 1(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 adopts the same implementation method as that of Example 1, except that in step 3, (R)-3-methylmorpholine (113.5 mg, 1.1240 mmol, 1.2 eq) is replaced with (1r, 4r)-4-methoxycyclohexane-1-amine (145.0 mg, 1.1240 mmol, 1.2 eq) to obtain 196 mg of an off-white solid (2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)-N-((1r, 4r)-4-methoxycyclohexyl)acetamide) with a yield of 56.0%. 1 HNMR(400MHz,DMSO-d6)δ10.15(s,1H),7.78(d,J=7.3Hz,1H),7.13(s,1H),6 .73(s,1H),3.83(s,3H),3.57(dd,J=11.6,6.2Hz,1H),3.47(dd,J=15.0,9.0 Hz,1H),3.39(s,2H),3.18(s,3H),2.30(s,3H),1.92(d,J=10.0Hz,2H),1.73 (d, J=10.4Hz, 2H), 1.11 (dd, J=25.0, 12.2Hz, 4H); ESI-MS: m / z=376.17[M+H] + .

[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 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 that of 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]heptane-6-yl)ethyl)-2H-chromen-2-one) with 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]

[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 by 7-hydroxy-6-methoxy-4-methyl-3-(2-oxo-2-(2-oxa-6-azaspiro[3.3]heptane-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 that of 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) with a yield of 63.4%. 1 HNMR(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 by 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 obtain 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 that of 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) with 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 by 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 that of 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) with a yield of 67.7%. 1H 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 by 7-hydroxy-6-methoxy-4-methyl-3-(2-oxo-2-((3aR, 6aS)-tetrahydro-1H-furano[3,4-c]pyrrol-5(3H)-yl)ethyl)-2H-chromen-2-one (124.0 mg, 0.3454 mmol, 1.0 eq) to obtain 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 adopts the same implementation method as that of Example 1, except that in step 3, (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) to obtain 183.5 mg of an off-white solid (3-(2-((1R, 4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-2-oxoethyl)-7-hydroxy-6-methoxy-4-methyl-2H-chromen-2-one) with a yield of 56.9%.1 H NMR (400MHz, DMSO-d6) δ10.17(s,1H),7.14(s,1H),6.74(s,1H),4.61(d,J=39.4Hz,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] 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-((1R, 4R)-2-oxa-5-azabicyclo[2.2.1]heptane-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 28.3 mg of a yellow-green solid (compound I-17), with a yield of 21.9%. 1 HNMR (400MHz, DMSO-d6)11.78(s,1H),10.37(s,1H),7.39(s,1H),4.69(t,J=53.8Hz,2H),3.88(s, 3H),3.83-3.16(m,6H),2.33(d,J=5.6Hz,3H),1.80(d,J=39.8Hz,2H); ESI-MS:m / z=396.11[M+Na] + .

[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 that of 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) with 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]heptane-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%. 1 HNMR(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 that of 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]decane-8-yl)ethyl)-2H-chromen-2-one) with 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%. 1H 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 that of 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) with 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 by 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 that of 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), and 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) with 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 by 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 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 that of 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), and 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 that of 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), and 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 5-(2-(8-formyl-7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetylamino)nicotinic acid methyl ester (Compound I-24)

[0134] This example adopts the same implementation method as that of 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), and 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)acetylamino)nicotinate methyl ester) with 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 by 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 adopts the same implementation 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)nicotinic acid methyl ester (50.0 mg, 0.1238 mmol) is added to 2 mL of 4 mol / L NaOH solution at one time and stirred at room temperature for 2 h. The reaction mixture is acidified to pH = 3 with 1 mol / L HCl. The precipitate formed is filtered, filtered with ethyl acetate and vacuum dried to obtain 11.5 mg of a brown solid (Compound I-25), yield: 23.8%.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 that of 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), and 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) with 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 that of 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), and 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) with 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(400MHz,DMSO-d6)δ11.82(s,1H),10.50(s,1H),10.40(s,1H),8.58(s,1H),8.02(d,J=8.8H z,1H),7.43(d,J=10.0Hz,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 adopts the same implementation method as that of 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-chloropyridine (144.5 mg, 1.1240 mmol, 1.2 eq), and the reaction time is 5 h to obtain 207.4 mg of an off-white solid (N-(5-chloropyridin-2-yl)-2-(7-hydroxy-6-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)acetamide) with a yield of 59.2%. 1 HNMR (400MHz, DMSO-d6) δ10.83(s,1H),10.20(s,1H),8.35(s,1H),8.02(d,J=8.8Hz,1H),7.85(d,J=8.8Hz, 1H),7.14(d,J=12.2Hz,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] 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-2-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 37.9 mg of dark green solid (compound I-28), yield: 28.7%. 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 proliferation inhibition activity of the compounds of the present invention on Mlg2908 cells and H358 cells was determined by Cell Counting Kit-8. Three replicate wells were set for each concentration of the test compound, and a blank control group with only culture medium but no cells and a negative control group with cells inoculated but no drug were set. Three replicate wells were set for both groups. After digestion of cells in the logarithmic growth phase, single cell suspension was blown into a 96-well culture plate; 5×10 cells were used for both Mlg2908 and H358 cells. 3 Cells / well. Add 100 μL of complete culture medium to each well and culture in an incubator with 5% carbon dioxide 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 three-fold diluted 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 an incubator for 3 hours. Detect the OD value of each well with an enzyme reader (detection wavelength: 450 nm), and calculate the inhibition rate 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 Proliferation inhibition activity of the compounds of the present invention on 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 inhibition activity than Orin1001, and have good application prospects.

Claims

1. A polysubstituted coumarin compound, the structural formula of which is as follows: in, R1 is selected from 2. A free form and a pharmaceutically acceptable salt or solvate of a polysubstituted coumarin compound according to claim 1.

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 and intrahepatic bile duct cancer, hepatocellular carcinoma, gastric cancer, glioma, glioblastoma, endometrial cancer, melanoma, kidney and renal pelvic 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 or villous colon adenoma.

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

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