Application of 8-amino-3-hydroxy-6H-benzo [c] chromene-6-ketone hydroxyl etherified derivative in preparation of medicine

Abstract

The invention belongs to the field of medicinal chemistry, and particularly relates to an application of an 8-amino-3-hydroxy-6H-benzo [c] chromene-6-ketone hydroxyl etherified derivative in preparation of drugs. The 8-amino-3-hydroxy-6H-benzo [c] chromene-6-ketone etherified derivative is as shown in the specification. According to the present invention, the acetamide protection group of the N-(3-hydroxy-6-oxo-6H-benzo [c] chromene-8-yl) acetamide hydroxyl etherification derivative is removed, the deprotection and purification process is optimized, and the biological activity detection results show that the obtained compound has good PDE2 inhibition activity, and the PDE2 inhibition activity is significantly improved by finely modifying the structure of the 8-site hydroxyurolithin A derivative; not only is the biological activity strong, but also the LogP value is obviously optimized. The optimization overcomes the pharmacokinetic defect of the active compound possibly caused by overhigh LogP in the prior art, and lays a solid foundation for developing a novel central nervous system disease treatment medicine with an excellent clinical application prospect. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and specifically relates to an 8-amino-3-hydroxy-6H-benzo[c]chromene-6-one hydroxyether derivative prepared from an N-(3-hydroxy-6-oxo-6H-benzo[c]chromene-8-yl)acetamide hydroxyether derivative as a raw material, which is used as an inhibitor of phosphodiesterase II (PDE2). Background Art

[0002] Cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) are ubiquitous second messengers in intracellular signal transduction pathways and play a crucial role in the treatment of central nervous system diseases. Phosphodiesterases (PDEs) are a multigene superfamily of enzymes encompassing 11 gene families. PDEs hydrolyze cAMP and / or cGMP, reducing their intracellular concentrations.

[0003] Phosphodiesterase II (PDE2) is a dual-substrate specific enzyme with a molecular weight of approximately 105 kDa that can simultaneously catalyze the hydrolysis of cAMP and cGMP. Phosphodiesterase inhibitors (PDEIs) primarily inhibit the activity of PDEs, preventing the hydrolysis of cAMP and / or cGMP, thereby increasing the concentrations of these two second messengers within the cell, thereby achieving a balance and playing a therapeutic role.

[0004] PDE2 has long been considered an important target for treating a variety of diseases. It has shown excellent therapeutic properties in the fields of the nervous system, anti-aging, and cardiovascular diseases, and has become a research hotspot.

[0005] Patent CN202411623411.6, a class of 8-hydroxyurolithin A derivatives and their preparation methods and applications, also protects the related structures of 8-hydroxyurolithin A derivatives. The most active is compound F16, with an IC50 of 0.63. However, the LogP value of this F16 derivative is 4.86 (LogP value can be used to measure the lipophilicity and hydrophilicity of a drug. A larger LogP value indicates a more lipophilic drug; conversely, a smaller LogP value indicates a more hydrophilic drug). Central nervous system drugs generally need to be able to cross the blood-brain barrier, so they tend to have high fat solubility. Generally speaking, drugs with LogP values between 0 and 3 are more easily absorbed because they are both sufficiently lipid-soluble to penetrate biological membranes and sufficiently water-soluble to ensure distribution and transport within the body. The LogP value of central nervous system drugs is a key pharmacokinetic parameter that determines the drug's absorption, distribution, and ability to penetrate the blood-brain barrier. LogP values range from -2 to 5, and the ideal LogP value for the central nervous system should be around 3. The aforementioned compound F16, despite its significant activity, has a LogP value of 4.86. While such a high LogP value indicates good lipid solubility, it also suggests that F16 may be excessively lipophilic, which could seriously affect its bioavailability, its ability to effectively penetrate the brain in a therapeutically relevant manner, or its overall drug-like properties.

[0006] Therefore, there is a continuing need in the art for novel 8-hydroxyurolithin A derivatives that should not only have significant biological activity but also exhibit optimized physicochemical properties, especially LogP values that are more conducive to CNS drug development, thereby providing a better balance between efficacy and drug-like properties. Summary of the Invention

[0007] The present invention aims to address the above-mentioned limitations of the prior art and provides a novel structure of N-(3-hydroxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide PDE2 inhibitor. At the same time, the present invention also provides a method for preparing such compound, which is scientific, reasonable, simple and easy to prepare.

[0008] In order to achieve the purpose of the present invention, the technical solution adopted by the present invention is as follows:

[0009] The hydroxy-etherified derivative of 8-amino-3-hydroxy-6H-benzo[c]chromene-6-one as a PDE2 inhibitor of the present invention has the general formula (I):

[0010]

[0011] Wherein, R is one of a straight-chain alkyl group, a branched-chain alkyl group, a haloalkyl group, a thioether alkyl group, an aryl alkyl group, a cycloalkylalkyl group, a heterocyclic alkyl group, a cyclopentyl group, a cyclohexyl group, a saturated heterocyclic group, an aryl group, an alkyl-substituted aryl group, a halogen-substituted aryl group, and a trifluoromethyl-substituted aryl group.

[0012] Furthermore, the straight-chain alkyl group is one of methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, and n-heptyl;

[0013] The branched alkyl group is one of isopropyl, isobutyl, sec-butyl, tert-butyl, and neopentyl;

[0014] The haloalkyl group is one of 2,2-difluoroethyl, 2,2,2-trifluoroethyl, and 2-chloroethyl;

[0015] The thioether alkyl group is 2-(methylthio)ethyl; the aryl alkyl group is benzyl; the cycloalkyl alkyl group is one of cyclopropylmethyl and cyclobutylmethyl; the heterocyclyl alkyl group is one of 2-furylmethyl and (tetrahydrofuran-2-yl)methyl; the saturated heterocyclyl group is tetrahydropyran-4-yl; the aryl group is phenyl; the alkyl-substituted aryl group is p-tolyl; the halogen-substituted aryl group is one of p-chlorophenyl, 3,4-dichlorophenyl, 2,4-dichlorophenyl, p-bromophenyl, and 2,4,6-tribromophenyl; and the trifluoromethyl-substituted aryl group is one of p-trifluoromethylphenyl and 3,5-bis(trifluoromethyl)phenyl. The specific structural formula of the N-(3-hydroxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide PDE2 inhibitor is as follows:

[0016]

[0017]

[0018] The preparation method of the hydroxy-etherified derivative of 8-amino-3-hydroxy-6H-benzo[c]chromene-6-one as a PDE2 inhibitor of the present invention comprises the following steps:

[0019]

[0020] The preferred steps are as follows:

[0021] Add N-(3-hydroxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide hydroxy ether derivative and concentrated hydrochloric acid to an anhydrous ethanol solution. Specially, if the substituent is not benzyl and its derivatives, add zinc chloride and reflux at 90°C for 4-12 hours. After the reaction, evaporate the solvent under reduced pressure, dissolve the remaining solid with ethyl acetate, wash with saturated sodium bicarbonate three times, and separate. The organic phase is spin-dried. Specially, if the substituent is not benzyl and its derivatives, recrystallize with methanol; specially, if the substituent is benzyl and its derivatives, obtain a solid product by column chromatography.

[0022] In the steps, the molar ratio of the N-(3-hydroxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide hydroxyether derivative to concentrated hydrochloric acid and zinc chloride is 2:2:1.

[0023] In the step, the column chromatography fluidity is dichloromethane and ammonia methanol, and the volume ratio is 100:1

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] The present invention provides PDE2 inhibitor compounds, primarily comprising 33 compounds of hydroxyether derivatives of 8-amino-3-hydroxy-6H-benzo[c]chromene-6-one, which exhibit excellent PDE2 inhibitory activity. These compounds have the potential to treat neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Huntington's disease, and can be used as active ingredients in the preparation of drugs that inhibit PDE2 activity.

[0026] These derivatives not only maintain excellent biological activity but, more importantly, exhibit optimized LogP values that are more conducive to CNS drug development. Among the series of compounds provided by this invention, the representative compound UE29 not only exhibits breakthrough nanomolar bioactivity, but also has an optimized LogP value of 3.71. This invention successfully maintains biological activity comparable to the best prior art compound (F16) while significantly improving the key physicochemical parameter, the LogP value. This makes compound UE29 even more promising as a CNS drug candidate, achieving an ideal balance between potent pharmacological effects and good drugability.

[0027] Furthermore, a corresponding preparation method is presented for the provided compound structure. Through process exploration, improved preparation and purification methods were developed to obtain a pure compound. The synthesis method is simple and offers high yields. Among these, the research focused on methods for removing the protecting group. Two commonly used methods for removing the acetyl group are p-toluenesulfonic acid and Lewis acid in an alcoholic solution. Experimental verification indicated that the former method resulted in high impurities and very low yields after the reaction, so it was not considered. The latter method uses simpler conditions and readily available raw materials. When the substituents are non-benzyl groups and their derivatives, simple washing and recrystallization are sufficient to obtain a pure product. However, when the substituents are benzyl groups and their derivatives, a significant number of byproducts are generated. After systematic analysis, cost-effectiveness studies, and subsequent purification considerations, it was ultimately decided to omit the ZnCl2 catalyst during the reaction to slow the reaction rate and prevent ether bond cleavage, which would result in a partial loss of conversion to ensure product purity. Furthermore, when the substituents are benzyl groups and their derivatives, column chromatography was chosen to separate the unreacted raw materials for purification. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 is the liquid phase purity of UE29.

[0029] Figure 2 This is a comparative experiment diagram of toxicity between F16 and UE29. DETAILED DESCRIPTION

[0030] The technical solutions of the present invention are described in detail below with reference to the following examples. The following examples are provided to illustrate, but not to limit, the synthesis methods of the compounds of Formula I. In one embodiment, the hydroxy-etherified derivatives of 8-amino-3-hydroxy-6H-benzo[c]chromene-6-one include the following compounds:

[0031] Example 1

[0032]

[0033] N-(3-Methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide (1 mmol, 0.28 g) and hydrochloric acid (1 mmol, 0.1 g) were added to anhydrous ethanol (20 mL) solution, and zinc chloride (0.5 mmol, 0.07 g) was added. The mixture was refluxed at 90°C for 8 h. After the reaction, the solvent was evaporated under reduced pressure, and the remaining solid was dissolved in ethyl acetate, washed with saturated sodium bicarbonate (3×20 mL) and separated. The organic phase was dried and recrystallized from methanol to obtain a yellow solid product 8-amino-3-methoxy-6H-benzo[c]chromen-6-one (Compound UE1).

[0034] UE1: yellow solid (75% yield), mp 246.6-248.6 ° C, 1 H NMR (400MHz, DMSO-d6) δ (ppm) 8.02 (dd, J=13.9, 8.6Hz, 2H), 7.34 (d, J=2.5Hz, 1H),7.14(dd,J=8.6,2.6Hz,1H),6.96-6.89(m,2H),5.81(s,2H),3.83(s,3H). 13 C NMR (101MHz, DMSO-d6) δ (ppm) 160.91, 159.21, 150.23, 149.05, 123.07, 122.97, 122.21, 120.25, 111.98, 111.79, 111.05, 101.31, 55.59.

[0035] Example 2

[0036]

[0037] Example 2 differs from Example 1 in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced with N-(3-ethoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 1, and a solid product UE2 is obtained in a yield of 84%.

[0038] UE2: yellow solid (84% yield), mp 207.4-208.9 ° C, 1 HNMR (400MHz, DMSO-d6) δ (ppm) 7.99 (dd, J=8.6, 6.3Hz, 2H), 7.35 (d, J=2.6Hz, 1H), 7.15 (dd, J= 8.8,2.5Hz,1H),6.93-6.86(m,2H),5.81(s,2H),4.08(q,J=6.9Hz,2H),1.35(t,J=6.9Hz,3H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)160.93,158.44,150.19,149.00,123.01,122.90,122.21,120.22,112.24,111.64,111.05,101.67,63.54,14.50.

[0039] Example 3

[0040]

[0041] Compared with Example 1, Example 3 differs in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced with N-(6-oxo-3-propoxy-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as in Example 1, and a solid product UE3 is obtained with a yield of 77%.

[0042] UE3: yellow solid (77% yield), mp 196.7-198.8 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.01(t,J=9.0Hz,2H),7.35(d,J=2.9Hz,1H),7.14(dd,J=8.6,2.6Hz,1H ),6.96-6.88(m,2H),5.81(s,2H),3.99(t,J=6.5Hz,2H),1.75(h,J=7.1Hz,2H),1.00(t,J=7.4Hz,3H). 13CNMR(101MHz,DMSO-d6)δ(ppm)160.92,158.61,150.21,148.94,123.06,123.01 ,122.90,122.24,120.22,112.27,111.65,111.10,101.73,69.35,21.91,10.35.

[0043] Example 4

[0044]

[0045] Example 4 differs from Example 1 in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced with N-(3-butoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 1, and a solid product UE4 is obtained in a yield of 68%.

[0046] UE4: yellow solid (68% yield), mp 173.0-175.4 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.04(d,J=8.7Hz,2H),7.42(d,J=2.4Hz,1H),7.21(dd,J=8.6,2.5Hz,1H), 6.97-6.88(m,2H),4.04(t,J=6.5Hz,2H),1.72(p,J=6.7Hz,2H),1.52-1.38(m,2H),0.95(t,J=7.4Hz,3H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)160.77,158.94,150.46,146.59,124.59,123.30,12 3.17,123.12,120.21,112.82,112.36,111.40,101.75,67.64,30.59,18.70,13.67.

[0047] Example 5

[0048]

[0049] Example 5 is different from Example 1 in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced with N-(6-oxo-3-(pentyloxy)-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as in Example 1, and a solid product UE5 is obtained with a yield of 71%.

[0050] UE5: yellow solid (71% yield), mp 163.6-165.2 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.01(dd,J=8.8,5.3Hz,2H),7.37(d,J=2.5Hz,1H),7.17(dd,J=8.6,2.5Hz,1H),6.95-6.86(m ,2H),5.85(s,2H),4.01(t,J=6.5Hz,2H),1.72(p,J=6.8Hz,2H),1.37(ddt,J=20.1,13.9,7.2Hz,4H),0.90(t,J=6.9Hz,3H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)160.90,158.64,150.23,148.74,123.20,123.01,122.9 2,122.33,120.23,112.29,111.63,111.26,101.72,67.89,28.25,27.66,21.88,13.90.

[0051] Example 6

[0052]

[0053] Compared with Example 1, Example 6 differs in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced by N-(3-(hexyloxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 1, and a solid product UE6 is obtained with a yield of 55%.

[0054] UE6: yellow solid (55% yield), mp 160.0-162.5 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.00(dd,J=7.9Hz,2H),7.36(d,J=2.4Hz,1H),7.16(dd,J=8.6,2.5Hz,1H),6.95-6.87(m,2 H),5.81(s,2H),4.01(t,J=6.5Hz,2H),1.71(p,J=6.8Hz,2H),1.41(p,J=7.2Hz,2H),1.34-1.25(m,4H),0.92-0.84(m,3H). 13CNMR(101MHz,DMSO-d6)δ(ppm)160.93,158.61,150.21,148.98,123.04,123.00,122.89,1 22.22,120.22,112.28,111.65,111.08,101.72,67.90,30.99,28.52,25.16,22.07,13.90.

[0055] Example 7

[0056]

[0057] Example 7 differs from Example 1 in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced with N-(3-isopropoxy-6-oxyl-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as in Example 1, and a solid product UE7 is obtained with a yield of 67%.

[0058] UE7: yellow solid (67% yield), mp 196.1-198.0 °C, 1 HNMR(400MHz,Chloroform-d)δ(ppm)7.78(dd,J=8.6,1.1Hz,2H),7.57(d,J=2.5Hz,1H),7.11(dd,J =8.5,1.5Hz,1H),6.87-6.79(m,2H),4.58(hept,J=6.0Hz,1H),4.05(s,2H),1.37(d,J=6.3Hz,6H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)161.37,157.97,150.85,148.60,124.08,123.55,123.10,120.71,113.64,112.19,111.96,103.28,70.17,22.16.

[0059] Example 8

[0060]

[0061] Compared with Example 1, Example 8 differs in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced by N-(3-isobutoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 1, and a solid product UE8 is obtained with a yield of 71%.

[0062] UE8: yellow solid (71% yield), mp 180.6-182.6 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.27(d,J=8.6Hz,1H),8.15(d,J=9.4Hz,1H),7.89(d,J=2.3Hz,1H),7.62(dd,J=8.6,2 .4Hz,1H),7.02-6.95(m,2H),5.77(s,2H),3.85(d,J=6.6Hz,2H),2.06(dq,J=13.2,6.6Hz,1H),1.01(d,J=6.7Hz,6H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)160.23,160.18,151.40,127.22,124.17,12 3.53,120.16,119.09,112.68,110.55,101.91,74.20,54.91,27.60,18.97.

[0063] Example 9

[0064]

[0065] Example 9 differs from Example 1 in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced with N-(3-(2-butoxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as in Example 1, and a solid product UE9 is obtained in a yield of 62%.

[0066] UE9: yellow solid (62% yield), mp 153.4-157.3 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.00(dd,J=8.7,6.3Hz,2H),7.36(d,J=2.5Hz,1H),7.16(dd,J=8.6,2.5Hz,1H),6.9 5-6.87(m,2H),5.81(s,2H),4.49(h,J=6.1Hz,1H),1.76-1.52(m,2H),1.26(d,J=6.0Hz,3H),0.94(t,J=7.4Hz,3H). 13CNMR(101MHz,DMSO-d6)δ(ppm)160.94,157.71,150.27,148.98,123.04,123.0 0,122.22,120.23,113.17,111.59,111.07,102.82,74.57,28.43,18.92,9.48.

[0067] Example 10

[0068]

[0069] Example 10 differs from Example 1 in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced with N-(3-(isopentyloxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 1, and a solid product UE10 is obtained in a yield of 55%.

[0070] UE10: yellow solid (55% yield), mp 160.0-163.2 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.17(d,J=8.7Hz,1H),8.10(d,J=8.7Hz,1H),7.70(d,J=2.4Hz,1H),7.46(dd,J=8.6,2.4Hz ,1H),7.00-6.91(m,2H),4.08(t,J=6.7Hz,2H),1.87-1.72(m,J=6.7Hz,1H),1.64(q,J=6.7Hz,2H),0.95(d,J=6.6Hz,6H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)160.47,159.58,151.00,125.48,123.72,12 3.36,120.18,116.32,112.57,110.92,101.82,66.50,37.24,24.54,22.39.

[0071] Example 11

[0072]

[0073] Example 11 differs from Example 1 in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced with N-(6-oxo-3-(pentan-2-yloxy)-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 1, and a solid product UE11 is obtained in a yield of 53%.

[0074] UE11: yellow solid (53% yield), mp 142.7-145.3 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.53-8.49(d,1H),8.22(d,J=8.7Hz,1H),8.11(d,J=8.4Hz,1H),8.00(dd,J=8.6,2.5Hz,1H),6.94(d,J=11.1H z,2H),5.81(s,2H),4.60-4.57(m,1H),1.69-1.62(m,1H),1.57-1.51( m,1H),1.42-1.33(m,2H),1.26(d,J=8.7Hz,3H),0.90(t,J=7.0Hz,3H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)160.48,159.16,151.47,138.98,129.68,126.05,124.1 8,122.70,119.61,118.06,113.33,110.46,102.68,73.34,37.80,19.36,18.09,13.85.

[0075] Example 12

[0076]

[0077] Example 12 differs from Example 1 in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced with N-(6-oxo-3-(pentane-3-oxy)-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as in Example 1, and a solid product UE12 is obtained in a yield of 72%.

[0078] UE12 yellow solid (72% yield), mp 125.5-126.7 ° C, 1 HNMR (400MHz, DMSO-d6) δ (ppm) 8.28 (d, J = 8.7Hz, 1H), 8.15 (d, J = 8.8Hz, 1H), 7.89 (d, J = 2.4Hz, 1H), 7.63 (dd, J = 8.6, 2.4Hz,1H),7.05-6.93(m,2H),4.40(p,J=5.8Hz,1H),1.65(ddq,J=14.3,7.3,3.6,3.2Hz,4H),0.92(t,J=7.4Hz,6H). 13CNMR(101MHz,DMSO-d6)δ(ppm)160.26,159.81,151.48,127.23,124.29,123.51,120.15,119.11,113.47,110.44,102.85,79.57,25.46,9.29.

[0079] Example 13

[0080]

[0081] Example 13 differs from Example 1 in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced with N-(3-(cyclopropanemethoxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as in Example 1, and a solid product UE13 is obtained with a yield of 75%.

[0082] UE13: yellow solid (75% yield), mp 190.6-193.2 °C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.01(d,J=6.7Hz,1H),7.99(d,J=5.8Hz,1H),7.36(d,J=2.5Hz,1H),7.15(dd,J=8.7,2.5Hz,1H),6.95- 6.88(m,2H),5.82(s,2H),3.88(d,J=7.0Hz,2H),1.24(pd,J=7.3,3.7Hz,1H),0.59(dt,J=8.1,3.0Hz,2H),0.35(dt,J=6.1,3.0Hz,2H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)160.92,158.56,150.19,148.98,123.02,12 2.89,122.22,120.22,112.30,111.63,111.07,101.80,72.51,10.01,3.13.

[0083] Example 14

[0084]

[0085] Example 14 differs from Example 1 in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced with N-(3-(cyclobutylmethoxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as in Example 1, and a solid product UE14 is obtained with a yield of 73%.

[0086] UE14: yellow solid (73% yield), mp 194.7-196.4 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.02(d,J=3.9Hz,1H),8.00(d,J=3.3Hz,1H),7.38(d,J=2.6Hz,1H),7.17(dd,J=8.6,2.5Hz,1H),6.9 5-6.88(m,2H),5.90(s,2H),4.01(d,J=6.7Hz,2H),2.73(hept,J=7.2Hz,1H),2.08(dtd,J=13.7,7.3,2.4Hz,2H),2.00-1.77(m,4H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)160.90,158.77,150.23,148.66,123.25,123.04,12 2.92,122.37,120.23,112.33,111.65,111.32,101.80,71.88,33.83,24.34,18.08.

[0087] Example 15

[0088]

[0089] Example 15 is different from Example 1 in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced with N-(3-(cyclopentylmethoxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as in Example 1, and a solid product UE15 is obtained with a yield of 60%.

[0090] UE15: yellow solid (60% yield), mp 200.4-204.6 ° C, 1HNMR(400MHz,DMSO-d6)δ(ppm)8.07(d,J=8.9Hz,1H),8.05(d,J=9.4Hz,1H),7.51(d,J=2.4Hz,1H),7.29(dd,J=8.6,2.5Hz,1H),6.96-6.89( m,2H),5.87(s,2H),3.91(d,J=7.0Hz,2H),2.32(hept,J=7.5Hz,1H),1.78(h,J=6.6Hz,2H),1.68-1.47(m,4H),1.34(dq,J=13.3,6.9Hz,2H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)160.72,159.18,150.56,125.15,123.69,123.26 ,123.17,120.21,113.43,112.44,111.33,101.81,72.05,38.39,28.95,24.91.

[0091] Example 16

[0092]

[0093] Example 16 differs from Example 1 in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced with N-(3-(cyclohexylmethoxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as in Example 1, and a solid product UE16 is obtained in a yield of 72%.

[0094] UE16: yellow solid (72% yield), mp 214.8-216.3 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.19(d,J=8.5Hz,1H),8.10(d,J=9.8Hz,1H),7.75(d,J=2.3Hz,1H),7.51(dd,J=8.7,2.4Hz,1H),6.95-6.89( m,2H),5.83(s,2H),3.85(d,J=6.2Hz,2H),1.78-1.72(m,2H),1.71(d,J=4.0Hz,1H),1.32-1.13(m,3H),1.06(pd,J=12.0,11.5,3.4Hz,2H). 13CNMR(101MHz,DMSO-d6)δ(ppm)160.39,159.85,151.10,128.44,126.02,123.84,12 3.39,120.14,117.19,112.55,110.76,101.83,73.16,36.92,29.14,25.99,25.21.

[0095] Example 17

[0096]

[0097] N-(3-((2-Fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide (1 mmol, 0.38 g) and hydrochloric acid (1 mmol, 0.1 g) were added to anhydrous ethanol (20 mL) solution, refluxed at 90°C, and reacted for 4 h. After the reaction, the solvent was evaporated under reduced pressure, and the remaining solid was dissolved in ethyl acetate, washed with saturated sodium bicarbonate (3×20 mL) and separated. The organic phase was spin-dried and purified by column chromatography (dichloromethane:ammonia methanol=100:1) to give the solid product 8-amino-3-(benzyloxy)-6H-benzo[c]chromen-6-one (compound UE17).

[0098] UE17: yellow solid (53% yield), mp 244.3-246.2 °C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.03(dd,J=17.3,8.8Hz,2H),7.60(td,J=7.6,1.7Hz,1H),7.49-7.42(m,1H),7.35(d,J=2.5Hz,1 H),7.31-7.23(m,2H),7.15(dd,J=8.6,2.5Hz,1H),7.08(d,J=2.6Hz,1H),7.01(dd,J=8.8,2.6Hz,1H),5.83(s,2H),5.22(s,2H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)161.35,159.73,158.50,150.62,149.60,131.42,131.09,125.06, 123.96,123.81,123.57,123.36,122.68,120.82,115.93,112.97,112.69,111.56,102.72,64.47.

[0099] Example 18

[0100]

[0101] Example 18 differs from Example 17 in that the N-(3-((2-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 17 is replaced with N-(3-((3-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 17, and a solid product UE18 is obtained in a yield of 43%.

[0102] UE18: yellow solid (43% yield), mp 236.5-239.4 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.02(dd,J=13.9,8.7Hz,2H),7.51-7.39(m,2H),7.37- 7.31(m,2H),7.20(td,J=8.4,2.9Hz,2H),7.07-6.98(m,2H),5.86(s,2H),5.23(s,2H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)163.88,161.40,158.39,150.59,149.59,140.07,130.96,124.08, 123.56,123.44,123.36,122.67,120.82,115.22,114.79,113.02,112.65,111.59,102.81,69.18.

[0103] Example 19

[0104]

[0105] Example 19 is different from Example 17 in that the N-(3-((2-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 17 is replaced by N-(3-((4-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 17 to obtain a solid product UE19 with a yield of 39%.

[0106] UE19: yellow solid (39% yield), mp 237.1-240.5 °C, 1HNMR(400MHz,DMSO-d6)δ(ppm)8.06(dd,J=16.1,8.8Hz,2H),7.50(td,J=8.2,6.2Hz,1H),7.43 -7.32(m,3H),7.22(ddd,J=13.1,8.1,2.3Hz,2H),7.10-7.01(m,2H),5.87(s,2H),5.26(s,2H). 13 C NMR (101MHz, DMSO-d6) δ (ppm) 163.88, 161.40 (d, J = 11.11Hz), 158.41, 150.60, 149.61, 140.08 (d, J = 7.07Hz), 130.99 (d, J = 8.08Hz), 12 4.10(d,J=3.03Hz),123.58,123.48,123.35,122.67,120.82,115.18(d,J=21.21Hz),114.81,113.06,112.66,111.56,102.82,69.19.

[0107] Example 20

[0108]

[0109] Example 20 differs from Example 17 in that the N-(3-((2-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 17 is replaced with N-(3-((3-methylbenzyl)oxy)-6-oxo-6H-benzo[c]coumarin-8-yl)acetamide. The other operations are the same as those in Example 17, and a solid product UE20 is obtained in a yield of 39%.

[0110] UE20: yellow solid (39% yield), mp 238.7-241.5 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.01(dd,J=14.0,8.7Hz,2H),7.36(d,J=2.5Hz,1H),7.28(dd,J=6.1,1.9Hz,3H),7.19-

[0111] 7.13(m,2H),7.04-6.97(m,2H),5.82(s,2H),5.14(s,2H),2.33(s,3H). 13CNMR(101MHz,DMSO-d6)δ(ppm)161.37,158.72,150.62,149.57,138.13,137.04,129.07,128 .85,125.39,123.56,123.43,122.69,120.78,113.09,112.48,111.56,102.74,70.14,21.45.

[0112] Example 21

[0113]

[0114] Example 21 differs from Example 17 in that the N-(3-((2-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 17 is replaced with N-(3-((4-methylbenzyl)oxy)-6-oxo-6H-benzo[c]coumarin-8-yl)acetamide. The other operations are the same as those in Example 17, and a solid product UE21 is obtained in a yield of 52%.

[0115] UE21: yellow solid (52% yield), mp 231.6-235.6 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.52(d,J=2.3Hz,1H),8.25(d,J=8.8Hz,1H),8.16(d,J=8.8Hz,1H),8.02(dd,J=8 .8,2.3Hz,1H),7.55(dd,J=8.2,5.5Hz,2H),7.25(t,J=8.9Hz,2H),7.11-7.01(m,2H),5.20(s,2H),2.11(s,3H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)163.07,160.42,159.48,151.34,139.13,132.73,130.22,130.13,129 .56,126.03,124.17,122.82,119.73,118.07,115.43,115.22,112.89,111.00,102.36,68.97,24.05.

[0116] Example 22

[0117]

[0118] Example 22 differs from Example 17 in that the N-(3-((2-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 17 is replaced with N-(3-((2-chlorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 17, and a solid product UE22 is obtained in a yield of 51%.

[0119] UE22: yellow solid (51% yield), mp 234.9-236.0 °C, 1 HNMR(400MHz, DMSO-d6)δ(ppm)8.05(d,J=8.9Hz,1H),8.00(d,J=8.8Hz,1H),7.66-7.59(m,1H),7.57-7.49(m,1H),7.46-7.37(m,2H ),7.34(d,J=2.6Hz,1H),7.14(dd,J=8.7,2.5Hz,1H),7.06(d,J=2.5Hz,1H),7.01(dd,J=8.8,2.6Hz,1H),5.82(s,2H),5.23(s,2H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)161.34,158.50,150.62,149.64,134.39,133.31,130.91,130.56, 129.94,127.90,123.62,123.54,123.33,122.67,120.83,112.95,112.78,111.55,102.80,67.82.

[0120] Example 23

[0121]

[0122] Example 23 differs from Example 17 in that the N-(3-((2-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 17 is replaced with N-(3-((3-chlorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 17, and a solid product UE23 is obtained in a yield of 46%.

[0123] UE23: yellow solid (46% yield), mp 235.2-238.4 ° C, 1HNMR (400MHz, DMSO-d6) δ (ppm) 8.10 (d, J = 8.7Hz, 1H), 8.06 (d, J = 8.7Hz, 1H), 7.61 (d, J = 1.9Hz, 1H), 7.53-7. 46(m,3H),7.39(d,J=2.5Hz,1H),7.20(dd,J=8.7,2.5Hz,1H),7.11-7.05(m,2H),5.87(s,2H),5.27(s,2H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)161.34,158.39,150.61,149.62,139.74,133.63,130.88,128.35, 127.89,126.77,123.59,123.49,123.35,122.67,120.83,113.06,112.69,111.57,102.84,69.12.

[0124] Example 24

[0125]

[0126] Example 24 differs from Example 17 in that the N-(3-((2-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 17 is replaced with N-(3-((4-chlorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 17, and a solid product UE24 is obtained in a yield of 45%.

[0127] UE24: yellow solid (45% yield), mp 266.3-268.1 °C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.01(dd,J=17.2,8.8Hz,2H),7.53-7.45(m,4H),7.33(d, J=2.5Hz,1H),7.13(dd,J=8.6,2.5Hz,1H),7.05-6.95(m,2H),5.81(s,2H),5.19(s,2H). 13 C NMR(101MHz,DMSO-d6)δ(ppm)161.34,158.46,150.60,149.61,136.21,133.01,130.13,1 28.97,123.61,123.50,123.35,122.68,120.79,113.12,112.63,111.53,102.84,69.21.

[0128] Example 25

[0129]

[0130] Example 25 differs from Example 17 in that the N-(3-((2-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 17 is replaced with N-(6-oxo-3-((3-(trifluoromethyl)phenyl)oxy)-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 17, to give a solid product UE 25 in a yield of 42%.

[0131] UE25: yellow solid (42% yield), mp 192.9-194.2 °C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)7.98(dd,J=15.6,8.8Hz,2H),7.83(d,J=1.9Hz,1H),7.78(d,J=7.6Hz,1H),7.69(d,J=8.0Hz, 1H),7.63(t,J=7.7Hz,1H),7.36(d,J=2.5Hz,1H),7.15(dd,J=8.7,2.5Hz,1H),7.06-6.96(m,2H),5.82(s,2H),5.27(s,2H). 13 C NMR(101MHz,DMSO-d6)δ(ppm)161.35,158.36,150.60,149.61,138.66,132.21,129.94,129.54,128. 68,125.98,125.11,124.60,123.51,123.30,122.66,120.83,113.00,112.72,111.59,102.80,69.15.

[0132] Example 26

[0133]

[0134] Example 26 differs from Example 17 in that the N-(3-((2-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 17 is replaced with N-(6-oxo-3-((4-(trifluoromethyl)phenyl)oxy)-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 17, to give a solid product UE 26 in a yield of 30%.

[0135] UE26: yellow solid (30% yield), mp 267.3-269.8 ° C,1 HNMR(400MHz,DMSO-d6)δ(ppm)8.05(d,J=8.8Hz,1H),8.00(d,J=8.7Hz,1H),7.78(d,J=8.1Hz,2H),7.70(d,J =8.1Hz,2H),7.33(d,J=2.5Hz,1H),7.13(dd,J=8.7,2.6Hz,1H),7.08-6.96(m,2H),5.81(s,2H),5.31(s,2H). 13 C NMR(101MHz,DMSO-d6)δ(ppm)161.31,158.32,150.60,149.63,142.06,129.03,128.71,128. 56,125.84,123.55,123.30,122.65,120.83,113.04,112.75,111.57,102.86,69.13,29.45.

[0136] Example 27

[0137]

[0138] Example 27 differs from Example 17 in that the N-(3-((2-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 17 is replaced with N-(3-((4-bromobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 17, and a solid product UE27 is obtained in a yield of 29%.

[0139] UE27: yellow solid (29% yield), mp 275.6-276.9 ° C, 1 HNMR (400MHz, DMSO-d6) δ (ppm) 8.02 (dd, J = 18.4, 8.7Hz, 2H), 7.78 (d, J = 7.9Hz, 2H), 7.71 (d, J = 8.1Hz, 2H),7.36(d,J=2.5Hz,1H),7.16(dd,J=8.6,2.6Hz,1H),7.06-6.99(m,2H),5.83(s,2H),5.32(s,2H). 13CNMR(101MHz,DMSO-d6)δ(ppm)161.31,158.32,150.60,149.63,142.06,128.56,125.85, 125.81,123.59,123.51,123.30,122.65,120.82,113.04,112.74,111.56,102.85,69.12.

[0140] Example 28

[0141]

[0142] Example 28 differs from Example 17 in that the N-(3-((2-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 17 is replaced with N-(3-((4-(tert-butyl)benzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 17, to give a solid product UE 28 in a yield of 31%.

[0143] UE28: yellow solid (31% yield), mp 232.9-235.1 °C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.01(dd,J=15.8,8.8Hz,2H),7.44-7.37(m,4H),7.33(d,J=2.5 Hz,1H),7.13(dd,J=8.7,2.5Hz,1H),7.04-6.95(m,2H),5.79(s,2H),5.14(s,2H),1.28(s,9H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)161.36,158.75,150.93,150.63,149.57,140.78,135.46,134.15,128.23 ,125.70,123.57,123.46,123.42,122.68,120.77,113.11,112.45,111.54,102.75,69.90,34.78,31.58.

[0144] Example 29

[0145]

[0146] Example 29 differs from Example 17 in that the N-(3-((2-fluorobenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 17 is replaced with N-(3-((3,5-dimethoxybenzyl)oxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 17, to give a solid product UE 29 in a yield of 39%.

[0147] UE29: yellow solid (39% yield), mp 188.6-190.2 °C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.50(d,J=2.3Hz,1H),8.22(d,J=8.6Hz,1H),8.14(d,J=9.5Hz,1H),8.00(dd,J=8.8,2.3H z,1H),7.03(dq,J=5.5,2.5Hz,2H),6.63(d,J=2.3Hz,2H),6.45(t,J=2.2Hz,1H),5.81(s,2H),5.19(s,2H),3.74(s,6H). 13 C NMR(101MHz,DMSO-d6)δ(ppm)161.05,160.89,159.98,151.80,139.61,139.32,130.05,126 .50,124.63,123.28,120.22,118.55,113.38,111.47,106.00,102.87,99.98,70.03,55.66.

[0148] Example 30

[0149]

[0150] Example 30 is different from Example 1 in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced by N-(3-(2-(methylthio)ethoxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide.

[0151] The other operations were the same as in Example 1 to obtain a solid product UE30 with a yield of 71%.

[0152] UE30: yellow solid (71% yield), mp 178.3-180.0 ° C, 1HNMR (400MHz, DMSO-d6) δ (ppm) 8.06-7.97 (m, 2H), 7.36 (d, J = 2.3Hz, 1H), 7.19-7.12 (m, 1H) ,6.98-6.90(m,2H),5.81(s,2H),4.22(t,J=6.6Hz,2H),2.88(t,J=6.5Hz,2H),2.18(s,3H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)160.89,158.11,150.17,123.05,122.95,122.21,120.27,112.31 ,111.94,111.08,101.91,67.34,40.09,39.88,39.68,39.47,39.26,39.05,38.84,32.05,15.20.

[0153] Example 31

[0154]

[0155] Compared with Example 1, Example 31 differs in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced by N-(3-(2-methoxyethoxy)-6-oxo-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 1, and a solid product UE31 is obtained with a yield of 67%.

[0156] UE31: yellow solid (37% yield), mp 180.1-183.1 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)8.01(t,J=8.7Hz,2H),7.36(d,J=2.4Hz,1H),7.16(dd,J=8.7,2 .5Hz,1H),6.98-6.90(m,2H),5.82(s,2H),4.20-4.14(m,2H),3.72-3.65(m,2H),3.33(s,3H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)160.90,158.38,150.17,149.05,123.05,122 .95,122.20,120.27,112.30,111.84,111.06,101.80,70.23,67.33,58.14.

[0157] Example 32

[0158]

[0159] Compared with Example 1, Example 32 differs in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced by N-(6-oxo-3-((tetrahydrofuran-2-yl)methoxy)-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as in Example 1 to obtain a solid product UE32 with a yield of 79%.

[0160] UE32: yellow solid (79% yield), mp 180.6-182.6 ° C, 1 HNMR(400MHz,DMSO-d6)δ(ppm)10.31(s,1H),8.15(d,J=8.9Hz,1H),8.13-8.07(m,1H),7.54(d,J=2.7Hz,1H),7.34(dd,J=8.7,2.7Hz,1H),6.97 (d,J=7.6Hz,2H),6.08(ddt,J=17.3,10.5,5.3Hz,1H),5.46(dq,J=17.3,1.7Hz,1H),5.31(dq,J=10.5,1.5Hz,1H),4.66(dt,J=5.3,1.6Hz,2H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)160.62,160.07,151.54,139.27,129.79,126.20,124.28,122.95,119.86,118.2 3,112.80,110.96,102.10,76.50,70.78,67.67,40.29,40.09,39.88,39.67,39.46,39.25,39.04,27.76,25.37.

[0161] Example 33

[0162]

[0163] Compared with Example 1, Example 33 differs in that the N-(3-methoxy-6-oxo-6H-benzo[c]chromen-8-yl)acetamide in Example 1 is replaced by N-(6-oxo-3-((tetrahydro-2H-pyran-4-yl)methoxy)-6H-benzo[c]chromen-8-yl)acetamide. The other operations are the same as those in Example 1 to obtain a solid product UE33 with a yield of 66%.

[0164] UE33: yellow solid (66% yield), mp 229.1-231.1 °C, 1HNMR (400MHz, DMSO-d6) δ (ppm) 8.00 (t, J = 7.7Hz, 2H), 7.36 (d, J = 2.4Hz, 1H), 7.15 (dd, J = 8.6, 2.5Hz, 1H), 6.96-6.87 (m, 2H), 5.82 (s, 2H) ,3.89(d,J=6.7Hz,4H),3.33(t,J=11.2Hz,2H),2.01(ddq,J=11.6,8.5,4.3Hz,1H),1.68(d,J=12.5Hz,2H),1.34(qd,J=12.2,4.4Hz,2H). 13 CNMR(101MHz,DMSO-d6)δ(ppm)160.91,158.57,150.19,149.03,123.03,122.99,12 2.91,122.20,120.24,112.30,111.75,111.06,101.82,72.37,66.58,34.30,29.14.

[0165] Effect example: Toxicity comparison experiment of F16 and UE29

[0166] In this study, the toxicity of F16 and UE29 from CN202411623411.6 was investigated using Caenorhabditis elegans. Synchronized L4 nematodes were cultured at 20°C. The positive control group consisted of 25 μM F16 (optimal concentrations were selected from 10-100 μM). Counts were taken from the moment of transfer, with the day of transfer designated as day 0 of the lifespan experiment. The number of nematodes surviving, dying, and missing was recorded daily to determine the average lifespan of each group, and the data were analyzed for survival. For detailed methods, see Medicinal Chemistry Research (2023) 32:1087–1097.

[0167] from Figure 1 As can be seen in the figure, UE29 shifted the survival curve to the right, extending the lifespan of the nematodes. This example shows that at a dose of 25 μM, UE29 significantly increased the lifespan of the nematodes compared to F16 and was less toxic than F16.

[0168] Effect example: Phosphodiesterase inhibitory activity study of the obtained product UE1-33

[0169] Experimental method: The inhibitor to be tested was dissolved in DMSO to a 1 μM dilution. BAY 60-7550 was removed from a -20°C freezer and diluted with DMSO to a 10 mM solution, followed by dilution with 1× Reaction Buffer (RA) to a gradient of 50 nM, 25 nM, 12.5 nM, 6.25 nM, 3.12 nM, 1.56 nM, and 0.78 nM. PDE2 protein of a predetermined concentration was taken from -80°C and dissolved in a 2.44 mg / mL protein dilution. cAMP was diluted with RA to a 1 μM solution.

[0170] Select an area on a 384-well white plate. Then, pipette 1 μL of the inhibitor dilution to be tested and 1.5 μL of the PDE2 protein dilution into a 284-well white plate. Set up three replicate wells per group. For the Min well, add 2.5 μL of RA (no protein) to each well; for the Max well, add 1 μL of RA (1.5 μL of protein); for the Positive well, add 1 μL of BAY 60-7550 (1.5 μL of protein); and for the Compound (1 μL) to each well. Cover the plate with sealing film and centrifuge at 1000 rpm for 1 minute. After centrifugation, place the plate in a thermostatted shaker at 25°C, 300 rpm, and incubate for 30 minutes. After the reaction, add 2.5 μL of cAMP solution to each well, centrifuge at 1000 rpm for 1 minute, and then incubate at 25°C, 300 rpm, and incubate for 20 minutes. Simultaneously remove the Termination buffer (T) and Detection buffer (D) from the refrigerator. After the reaction is complete, add 2.5 μL of T to each well and centrifuge under the same conditions. Then, add 2.5 μL of D to each well and centrifuge under the same conditions for 20 minutes. After the reaction is complete, add Kinase Reagent buffer (KR) in the dark, cover with aluminum foil, centrifuge under the same conditions, and continue incubation for another 10 minutes. During the reaction, open the multifunctional microplate reader, select the program, select the area, and after the reaction is complete, place the white plate in the instrument's detection area, run the program, and obtain the results.

[0171] Only the inhibitor dilution solution was replaced with BAY 60-7550 dilution solution of each concentration gradient. The reagents were added in sequence according to the above experimental process. The data were read using a microplate reader and the IC was calculated using GraphPad Prism 8. 50 .

[0172] The IC values of the 33 synthesized compounds were further determined. 50 The potential compound was diluted with 1× reaction buffer to 7 final concentrations of 200μM, 100μM, 50μM, 25μM, 12.5μM, 6.25μM, and 3.125μM. The specific experimental steps refer to the IC value of BAY60-7550 mentioned above. 50 Determination method.

[0173] Table 1

[0174]

[0175] The logP values of the 33 synthesized compounds were further determined by using ChemRraw software to predict the compounds.

[0176] Table 2

[0177]

[0178]

[0179] From the specific IC in the table above 50 The data show that the hydroxy ether derivatives of the general formula 8-amino-3-hydroxy-6H-benzo[c]chromene-6-one exhibit excellent inhibitory effects on PDE2 and have the potential to treat central nervous system diseases such as memory impairment, cognitive impairment, anxiety and depression. They can be used as active ingredients to prepare drugs that inhibit PDE2 activity. Among them, except for compounds UE4, UE5, UE9, UE13, UE18, UE19, UE21, UE27 and UE28, this series of compounds IC 50 All <50, and the IC of UE29 50 The breakthrough reached the nanomolar level. The logP value of UE29 is 3.71, which is lower than the 4.86 of F16 in CN202411623411.6. The value of 3.71 is also a relatively ideal value.

[0180] At the same time, the purity of UE29 was determined, such as Figure 1 As shown, the purity was 100% using the HPLC method. Instrument: Agilent Technologies 1260 Infinity; Column: C18; Mobile phase: methanol: water 100:0;

[0181] Detection wavelength: 264nm.

[0182] Although the present invention has been described by way of specific embodiments, it should not be construed as being limited thereto; rather, the present invention encompasses the general aspects disclosed hereinabove. Various modifications and embodiments are possible without departing from the spirit and scope of the present invention.

[0183] The above description is only a preferred specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any technician familiar with the technical field, within the technical scope disclosed by the present invention, who makes equivalent replacements or changes based on the technical solutions and concepts of the present invention, should be covered by the scope of protection of the present invention.

Claims

1. Use of a hydroxy-etherified derivative of 8-amino-3-hydroxy-6H-benzo[c]chromene-6-one in the preparation of phosphodiesterase II inhibitors, characterized in that: The structural formula of the 8-amino-3-hydroxy-6H-benzo[c]chromene-6-one hydroxy ether derivative is as follows: Wherein, R is one of a straight-chain alkyl group, a branched-chain alkyl group, a haloalkyl group, a thioether alkyl group, an aryl alkyl group, a cycloalkylalkyl group, a heterocyclic alkyl group, a cyclopentyl group, a cyclohexyl group, a saturated heterocyclic group, an aryl group, an alkyl-substituted aryl group, a halogen-substituted aryl group, and a trifluoromethyl-substituted aryl group.

2. The hydroxyetherified derivative of 8-amino-3-hydroxy-6H-benzo[c]chromene-6-one as a phosphodiesterase II inhibitor according to claim 1, characterized in that: The derivative is one of the following structural formulas:

3. Use of the hydroxyetherified derivative of 8-amino-3-hydroxy-6H-benzo[c]chromen-6-one according to claim 1 in the preparation of phosphodiesterase II inhibitor drugs, characterized in that: The derivative has the following structural formula:

4. A pharmaceutical composition, characterized in that The invention comprises the compound according to claim 1 or a pharmaceutically acceptable salt, ester, prodrug or solvate thereof, and one or more pharmaceutically acceptable carriers or excipients.

5. The pharmaceutical composition according to claim 4, characterized in that The pharmaceutically acceptable carrier or excipient is selected from at least one of a filler, a binder, a disintegrant, a lubricant, a glidant, a coating material, a sweetener, a flavoring agent, a colorant, a preservative, an antioxidant, and a pH regulator.

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