Application of IDPA and homologues thereof in preparation of antiviral drugs

Abstract

The invention belongs to the field of medicines, and particularly relates to application of IDPA and homologues thereof in preparation of antiviral drugs. The invention firstly discovers that IDPA and homologues thereof can inhibit the replication of porcine reproductive and respiratory syndrome virus (PRRSV) in in-vitro cells, inhibit the expression of PRRSV-N protein, inhibit the replication level of PRRSV and reduce the virus titer of PRRSV, in-vitro cell pharmacological experiments show that IDPA and homologues thereof have exact and good antiviral effects, provide new drugs for prevention and treatment of PRRSV, and have broad application prospects. The traditional Chinese medicine has the characteristics of moderate dosage, remarkable curative effect, convenience in administration, small toxic and side effects and the like, and has wide application prospects.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and particularly relates to the application of IDPA and its homologues in the preparation of antiviral drugs. Background Art

[0002] Porcine reproductive and respiratory syndrome (PRRS) is one of the diseases that threatens the healthy development of the global pig industry, causing serious economic losses to the industry. PRRSV infection mainly causes reproductive disorders in pregnant sows, specifically manifested as reproductive disorders such as miscarriage, stillbirth, weak piglets and mummified fetuses, as well as respiratory diseases in pigs of all ages. PRRSV mutates rapidly, strains recombine frequently, cross-protection between strains is low, and the protective effect of existing vaccines is poor, resulting in the widespread existence of the disease in my country. In addition, PRRSV is a major component of the porcine respiratory disease complex (PRDC). When it co-infects the host with other bacteria or viruses, it often aggravates the clinical symptoms of pigs and increases the mortality rate. In pig herds infected with PRRSV, the presence of PRRSV will also lead to increased costs of vaccines and drugs for related diseases.

[0003] PRRSV remains one of the most important pathogens in global pig production. Vaccination is currently the primary preventive measure, but inactivated vaccines are ineffective, and attenuated vaccines carry a significant risk of spreading the virus. Currently, there is no specific treatment for this disease, and comprehensive prevention and control measures and symptomatic treatment are primarily employed. The clinical efficacy of chemical agents such as tilmicosin and tyvalosin remains to be further characterized. Therefore, there is an urgent need to develop highly effective anti-PRRSV drugs to prevent and control the occurrence and progression of porcine blue ear disease.

[0004] Due to the high frequency of mutation and super immune escape ability of PRRSV, vaccine research is at a bottleneck period. Therefore, finding new antiviral strategies has become a hot topic in the current research on the prevention and control of porcine blue ear disease. Summary of the Invention

[0005] In response to the above technical problems, the present invention has prepared IDPA and its homologues. After incubating MARC-145 cells infected with PRRSV with the IDPA and its homologues, the replication of PRRSV can be significantly inhibited. It is expected to be used in the development of drugs for preventing or treating PRRSV infection, providing new materials and solutions for the effective prevention and control of PRRS.

[0006] Specifically include the following:

[0007] In a first aspect, the present invention provides IDPA and its homologues, wherein the structural formula of the IDPA and its homologues is shown in the following formula (I):

[0008]

[0009] Wherein, Ar is an aryl group, A is a straight chain or a 5- to 7-membered ring, R is a hydrogen alkyl group or an aryl group, and R' is an alkyl group or an aryl group.

[0010] Preferably, the aryl group is selected from substituted or unsubstituted phenyl groups.

[0011] Preferably, the substituted phenyl group includes an alkoxyphenyl group, a halogenated alkylphenyl group, a cyanophenyl group, a halogenated phenyl group, and an alkylphenyl group.

[0012] Preferably, A is a straight chain, and R is a phenyl group.

[0013] Preferably, A is a 5- to 7-membered carbon ring or a 5- to 7-membered oxygen heterocycle, and R is hydrogen.

[0014] Preferably, the IDPA and its homologues are selected from the compounds shown below:

[0015]

[0016] In a second aspect, the present invention provides a use of the IDPA and its homologues described in the first aspect in the preparation of antiviral drugs.

[0017] Preferably, the virus is porcine reproductive and respiratory syndrome virus.

[0018] Preferably, the IDPA and its homologues inhibit the replication of porcine reproductive and respiratory syndrome virus.

[0019] Preferably, the IDPA and its homologues inhibit the expression of PRRSV viral proteins.

[0020] Preferably, the IDPA and its homologues inhibit PRRSV replication.

[0021] Preferably, the IDPA and its homologues reduce PRRSV viral titer.

[0022] Preferably, the IDPA and its homologues or pharmaceutically acceptable salts thereof are added with pharmaceutically acceptable excipients to prepare any pharmaceutically acceptable dosage form.

[0023] Preferably, the dosage forms include tablets, sprays, granules, capsules, oral solutions, and injections.

[0024] The beneficial effects of the present invention are: first, the present invention prepares an IDPA and its homologues; second, the IDPA and its homologues can inhibit the expression of porcine reproductive and respiratory syndrome virus capsid protein, inhibit the viral titer of PRRSV, and inhibit the replication and proliferation of PRRSV, and in vitro cell efficacy tests show that they have a definite and good antiviral effect; the present invention provides a new drug for the prevention and treatment of PRRS, which has the characteristics of moderate dosage, significant efficacy, convenient medication, small toxic and side effects, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are merely embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying any creative work.

[0026] Figure 1 RT-qPCR detection results of the PRRSV ORF7 gene in MARC-145 cells infected with PRRSV SD-YL1712 strain after incubation with IDPA; the control group consisted of MARC-145 cells infected with PRRSV SD-YL1712 strain and treated with dimethyl sulfoxide (DMSO); the IDPA-treated group consisted of MARC-145 cells infected with PRRSV SD-YL1712 strain and treated with IDPA (10 μM);

[0027] Figure 2 RT-qPCR results of PRRSV ORF7 gene in MARC-145 cells infected with PRRSV SD-YL1712 strain after incubation with IDPA. The control group consisted of MARC-145 cells infected with PRRSV SD-YL1712 strain and treated with dimethyl sulfoxide (DMSO). The IDPA-treated group consisted of MARC-145 cells infected with PRRSV SD-YL1712 strain and treated with different doses of IDPA (2, 4, 6, 8, and 10 μM) in a dose-dependent manner.

[0028] Figure 3 RT-qPCR detection results of PRRSV ORF7 gene after incubation with 10 μM IDPA homolog in MARC-145 cells infected with PRRSV SD-YL1712 strain;

[0029] Figure 4Western blot analysis of PRRSV N protein in MARC-145 cells infected with PRRSV SD-YL1712 strain after incubation with IDPA. The inoculated control consisted of MARC-145 cells infected with PRRSV SD-YL1712 strain alone and treated with dimethyl sulfoxide (DMSO). The IDPA group consisted of MARC-145 cells infected with PRRSV SD-YL1712 strain and treated with IDPA. Figure 5 Virus titer in the cell culture supernatant of MARC-145 cells infected with PRRSV SD-YL1712 after incubation with IDPA. The inoculated control consisted of MARC-145 cells infected with PRRSV SD-YL1712 alone and treated with dimethyl sulfoxide (DMSO). The IDPA group consisted of MARC-145 cells infected with PRRSV SD-YL1712 and treated with IDPA. DETAILED DESCRIPTION

[0030] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0031] DMEM medium, trypsin, and fetal bovine serum were purchased from BI Company; β-actin monoclonal antibody was purchased from Beijing Quanshijin Biotechnology Co., Ltd.; dimethyl sulfoxide (DMSO) was purchased from Sigma-Aldrich Company; TRIzol for RNA extraction was purchased from Dalian Baoriyi Biological Co., Ltd.; reverse transcription kit and fluorescence quantitative PCR kit were purchased from Nanjing Novozymes Biotechnology Co., Ltd.; PRRSV ORF7 gene detection primers and internal reference gene GAPDH primers were synthesized by Beijing Qingke Biotechnology Co., Ltd.; MARC-145 cells (a derivative cell line of African green monkey kidney cells MA-104), highly pathogenic PRRSV strain SD-YL1712, and PRRSV-N protein monoclonal antibody were stored in the laboratory of the Animal Immunity and Metabolism Innovation Team of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences.

[0032] Example 1 Preparation of IDPA and its homologues

[0033] IDPA and its homologues (compounds 2-33) involved in this example are shown below:

[0034]

[0035] The specific synthesis method in this embodiment is:

[0036] Synthesis of Compound 2: 1-Indanone oxime (59.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 2 (IDPA, 181 mg, 85% yield) as a white solid. Product assay data are as follows:

[0037] 1 H NMR (400MHz, CDCl3): δ10.07(d,J=9.6Hz,1H),8.62(d,J=8.4Hz,1H),8.00(dd,J=12.4Hz,J=6.8Hz,4H),7.73(dd,J= 12.4Hz,J=6.8Hz,4H),7.57(t,J=7.2Hz,2H),7.52-7.47(m,4H),7.44-7.35(m,6H),7.26-7.16(m,3H),3.28(s,2H); 13 C NMR (100MHz, CDCl3): δ157.8(d,J C-P =5Hz),144.8(d,J C-P =12Hz),138.4(d,J C-P =13Hz),132.9(d,J C-P =118Hz),132.8(d,J C-P =114Hz),132.0(d,J C-P =3Hz),131.8(d,J C-P =3Hz),131.51(d,J C-P =10Hz),131.49(d,J C-P =10Hz),128.58(d,J C-P =12Hz),128.56(d,J C-P =12Hz),127.4,126.7,124.8,123.4(d,J C-P=2Hz),103.6(dd,J C-P =114Hz,J C-P =8Hz),37.9(d,J C-P =12Hz); 31 PNMR (243MHz, CDCl3): δ31.5,18.0; ESI-HRMS: m / z Calcd for C 33 H 27 NO2P2:531.1517,found531.1515.

[0038] Synthesis of Compound 3: 1-Indanone oxime (59.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (0.18 mL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed. Finally, a yellow solid product 3 (114 mg, yield 86%) was obtained. The product monitoring data are as follows:

[0039] 1 H NMR (400MHz, CDCl3): δ7.71 (dd, J=12.0Hz, J=6.8Hz, 4H), 7.53-7.40 (m, 6H), 7.38-7.30 (m, 4H), 5.72 (brs, NH2, 2H), 3.28 (d, J=2.0Hz, 2H); 13 C NMR (100MHz, CDCl3): δ158.8(d,J C-P =7Hz),145.2(d,J C-P =11Hz),139.4(d,J C-P =13Hz),134.4(d,J C-P =104Hz),131.5,131.4(d,J C-P =10Hz),128.4(d,J C-P =12Hz),127.6,126.4,124.1(d,J C-P =2Hz),118.2,91.1(d,JC-P =119Hz),38.0(d,J C-P =13Hz); 31 P NMR (243MHz, CDCl3): δ30.2; ESI-HRMS: m / z Calcd for C 21 H 18 NOP:331.1126, found 331.1124.

[0040] Synthesis of Compound 4: 1-Tetralone oxime (65.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain product 4 (109 mg, 50% yield) as a white solid. Product monitoring data are as follows:

[0041] 1 H NMR (400MHz, CDCl3): δ9.96(d,J=8.4Hz,1H),8.41(d,J=7.6Hz,1H),7.76-7.66(m,8H),7.58(t,J=7.2Hz,2H),7.50-7.46(m,4H),7.35(t,J= 7.2Hz,2H),7.29(d,J=7.6Hz,1H),7.26-7.21(m,4H),7.16(t,J=7.2Hz,1H),6.90(d,J=7.2Hz,1H),2.37(t,J=7.2Hz,2H),1.90-1.85(m,2H); 13 C NMR (100MHz, CDCl3): δ153.6,137.6,132.3(d,J C-P =113Hz),132.1(d,J C-P =3Hz),131.7(d,J C-P =104Hz),131.6(d,J C-P =10Hz),131.5(d,J C-P =9Hz),131.4(d,J C-P=3Hz),129.5,128.58(d,J C-P =12Hz),128.56,128.0(d,J C-P =13Hz),126.2,126.1,104.0(dd,J C-P =102Hz,J C-P =7Hz),28.0(d,J C-P =6Hz),25.5(d,J C-P =10Hz); 31 P NMR (243MHz, CDCl3): δ36.2,18.8; ESI-HRMS: m / z Calcd for C 34 H 29 NO2P2:545.1674, found 545.1670.

[0042] Synthesis of Compound 5: 1-Tetralone oxime (65.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (0.18 mL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed. Finally, a red solid product 5 (69 mg, 50% yield) was obtained. The product monitoring data are as follows:

[0043] 1 H NMR (400MHz, CDCl3): δ7.75 (dd, J=12.0Hz, J=7.2Hz, 4H), 7.52 (t, J=7.2Hz, 2H), 7.48-7.44 (m, 6H), 7. 29-7.26(m,1H),7.16(t,J=5.6Hz,1H),5.95(brs,NH2,2H),2.68(t,J=7.6Hz,2H),2.06-2.00(m,2H); 13 C NMR (100MHz, CDCl3): δ153.2(d,J C-P =4Hz),138.5(d,J C-P =2Hz),133.4(d,JC-P =102Hz),132.0(d,J C-P =10Hz),131.6(d,J C-P =10Hz),131.5(d,J C-P =2Hz),129.3,128.4(d,J C-P =11Hz),127.7,126.7,121.9,87.2(d,J C-P =108Hz),28.7(d,J C-P =7Hz),24.3(d,J C-P =11Hz); 31 P NMR (243MHz, CDCl3): δ37.9; ESI-HRMS: m / zCalcd for C 22 H 20 NOP:345.1283, found 345.1281.

[0044] Synthesis of Compound 6: 1-Benzocycloheptanone oxime (71.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed. The product 6 (107 mg, 48% yield) was obtained as a white solid. Product monitoring data are as follows:

[0045] 1 H NMR (400MHz, CDCl3): δ10.41(d,J=7.2Hz,1H),8.19-8.14(m,1H),8.00(d,J =7.2Hz,1H),7.90-7.87(m,2H),7.70-7.62(m,3H),7.58-7.46(m,10H),7.32 -7.25(m,2H),7.18-7.14(m,3H),6.74(d,J=7.6Hz,1H),1.90-1.86(m,1H), 1.82-1.66(m,2H),1.63-1.54(m,1H),1.51-1.42(m,1H),1.12-1.02(m,1H);13 C NMR (100MHz, CDCl3): δ156.5,140.4,131.9(d,J C-P =2Hz),131.8(d,J C-P =2Hz),131.52(d,J C-P =9Hz),131.5(d,J C-P =102Hz),131.42(d,J C-P =10Hz),131.4(d,J C-P =107Hz),129.6,128.7,128.6(d,J C-P =14Hz),128.5(d,J C-P =11Hz),127.7(d,J C-P =13Hz),127.2,125.4,100.0(dd,J C-P =100Hz,J C-P =7Hz),33.0,30.4,25.4(d,J C-P =11Hz); 31 P NMR (243MHz, CDCl3): δ38.2,15.8; ESI-HRMS: m / zCalcd for C 35 H 31 NO2P2:559.1830,found 559.1826.

[0046] Synthesis of Compound 7: 1-Benzocycloheptanone oxime (71.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (0.18 mL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed. Finally, a white solid product 7 (42 mg, yield 29%) was obtained. The product monitoring data are as follows:

[0047] 1H NMR (400MHz, CDCl3): δ7.74 (dd, J=11.6Hz, J=6.8Hz, 4H), 7.59-7.51 (m, 3H), 7.48-7.44 (m, 4H), 7.33-7.27 ( m,2H),7.17-7.15(m,1H),5.79(brs,NH2,2H),2.56(t,J=6.4Hz,2H),1.79-1.72(m,2H),1.42-1.32(m,2H); 13 C NMR (100MHz, CDCl3): δ159.0(d,J C-P =5Hz),140.4139.0(d,J C-P =14Hz),131.7(d,J C-P =11Hz),131.4(d,J C-P =2Hz),129.9(d,J C-P =139Hz),129.1,128.7,128.3(d,J C-P =11Hz),126.6,125.9,87.4(d,J C-P =106Hz),33.0,31.5,25.4(d,J C-P =11Hz); 31 P NMR (243MHz, CDCl3): δ38.2; ESI-HRMS: m / z Calcd forC 23 H 22 NOP:359.1439, found 359.1436.

[0048] Synthesis of Compound 8: 2-Methoxybenzocycloheptan-5-one oxime (83.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 8 (170 mg, 72% yield) as a white solid. Product monitoring data are as follows:

[0049] 1H NMR(400MHz,CDCl3):δ10.28(d,J=11.2Hz,1H),8.21-8.16(m,2H),7.96(d,J=7.2Hz,1H),7.92-7.87(m,2H),7.69-7.61(m,13H),7.30-7.27(m,1H),7.20-7.16(m,2H),6.85(dd,J=8.8Hz,J=2.8Hz,1H),6.30(d,J=2.8Hz,1H),3.78(s,3H),1.83-1.62(m,4H),1.52-1.42(m,1H),1.11-1.02(m,1H); 13 C NMR(100MHz,CDCl3):δ160.3,156.5(d,J C-P =3Hz),142.5,133.5(d,J C-P =102Hz),132.9,132.8(d,J C-P =100Hz),132.0(d,J C-P =3Hz),131.9(d,J C-P =3Hz),131.8(d,J C-P =10Hz),131.5(d,J C-P =9Hz),130.9(d,J C-P =3Hz),128.5(d,J C-P =12Hz),127.7(d,J C-P =13Hz),113.4,109.6,99.3(dd,J C-P =100Hz,J C-P =6Hz),55.1,32.8,30.6,25.6(d,J C-P =12Hz); 31 P NMR(243MHz,CDCl3):δ38.1,16.0;ESI-HRMS:m / z Calcd for C 36 H 33 NO3P2:589.1936,found 589.1934。

[0050] Synthesis of Compound 9: 5-Trifluoromethyl-1-indanone oxime (87.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed. The product 9 (177 mg, 74% yield) was obtained as a yellow solid. Product monitoring data are as follows:

[0051] 1 H NMR (400MHz, CDCl3): δ10.08(d,J=9.6Hz,1H),8.78(d,J=8.4Hz,1H),7.99(dd,J=12.8Hz,J=6.8H z,4H),7.72(dd,J=12.4Hz,J=6.8Hz,4H),7.61(t,J=7.2Hz,2H),7.55-7.38(m,12H),3.34(s,2H); 13 C NMR (100MHz, CDCl3): δ156.7(d,J C-P =8Hz),145.0(d,J C-P =11Hz),141.7(d,J C-P =13Hz),132.6(d,J C-P =127Hz),132.4(d,J C-P =2Hz),132.2(d,J C-P =127Hz),132.1(d,J C-P =3Hz),131.8,131.6(d,J C-P =10Hz),131.5(d,J C-P =11Hz),129.2(q,J C-F =32Hz),128.8(d,J C-P =12Hz),128.7(d,J C-P =13Hz),125.2,124.2(q,J C-F =271Hz),124.0(d,J C-P =4Hz),120.3,106.9(dd,J C-P=111Hz,J C-P =8Hz),38.0(d,J C-P =12Hz); 31 PNMR (162MHz, CDCl3): δ31.2,18.5; 19 F NMR (376MHz, CDCl3): δ-62.1; ESI-HRMS: m / zCalcd for C 34 H 26 F3NO2P2:599.1391, found 599.1386.

[0052] Synthesis of Compound 10: 5-Cyano-1-indanone oxime (70.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 10 (144 mg, 65% yield) as a white solid. Product monitoring data are as follows:

[0053] 1 H NMR (600MHz, CDCl3): δ10.09(d,J=9.6Hz,1H),8.82(d,J=8.4Hz,1H),7.98(dd,J=12.0Hz,J=6.6Hz,4H),7.72(dd,J=12.0Hz, J=6.6Hz,4H),7.62(t,J=7.2Hz,2H),7.55-7.53(m,5H),7.50(s,1H),7.47(t,J=7.2Hz,2H),7.42-7.39(m,4H),3.33(s,2H); 13 CNMR (150MHz, CDCl3): δ156.3(d,J C-P =6Hz),145.0(d,J C-P =11Hz),142.6(d,J C-P =12Hz),132.5(d,J C-P =3Hz),132.3(d,J C-P =126Hz),132.2(d,JC-P =3Hz),132.1(d,J C-P =127Hz),131.5(d,J C-P =10Hz),131.4(d,J C-P =10Hz),131.0,128.8(d,J C-P =12Hz),128.7(d,J C-P =12Hz),126.8,125.5,119.1,110.5,108.3(dd,J C-P =110Hz,J C-P =8Hz),37.9(d,J C-P =12Hz); 31 PNMR (162MHz, CDCl3): δ31.0,18.7; ESI-HRMS: m / z Calcd for C 34 H 26 N2O2P2:556.1470,found556.1467.

[0054] Synthesis of Compound 11: 5-Methyl-6-chloro-1-indanone oxime (78.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 11 (134 mg, 58% yield) as a white solid. Product monitoring data are as follows:

[0055] 1 H NMR (400MHz, CDCl3): δ10.02(d,J=10.0Hz,1H),8.69(s,1H),8.01(dd,J=12.4Hz,J=6.8Hz,4H),7.71(dd,J=12.4Hz ,J=7.2Hz,4H),7.58(t,J=7.2Hz,2H),7.53-7.48(m,4H),7.45-7.37(m,6H),7.08(s,1H),3.20(s,2H),2.29(s,3H); 13C NMR (100MHz, CDCl3): δ156.8(d,J C-P =7Hz),143.3(d,J C-P =12Hz),137.6(d,J C-P =15Hz),135.3,133.1,132.8(d,J C-P =114Hz),132.7(d,J C-P =119Hz),132.2(d,J C-P =3Hz),132.0(d,J C-P =3Hz),131.5(d,J C-P =10Hz),131.5(d,J C-P =10Hz),128.7(d,J C-P =12Hz),128.7(d,J C-P =12Hz),125.6,125.3,104.1(dd,J C-P =113Hz,J C-P =6Hz),37.4(d,J C-P =12Hz),20.3; 31 P NMR (162MHz, CDCl3): δ31.3,17.9; ESI-HRMS: m / z Calcd for C 34 H 28 ClNO2P2:579.1284,found579.1278.

[0056] Synthesis of Compound 12: 5-Fluoro-1-indanone oxime (66.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 12 (114 mg, 52% yield) as a white solid. Product monitoring data are as follows:

[0057] 1H NMR(400MHz,CDCl3):δ10.07(d,J=10.0Hz,1H),8.63(dd,J=9.6Hz,J=5.2Hz,1H),7.99(dd,J=12.4Hz,J=6.8Hz,4H),7.72(dd,J=12.4Hz,J=6.8Hz,4H),7.58(t,J=7.2Hz,2H),7.53-7.49(m,4H),7.46-7.36(m,6H),6.94(t,J=7.2Hz,2H),3.26(s,2H); 13 CNMR(100MHz,CDCl3):δ162.6(d,J C-F =247Hz),157.0(d,J C-P =4Hz),147.4(d,J C-P =11Hz,J C-F =9Hz),134.4(d,J C-F =13Hz),132.6(d,J C-P =123Hz),132.5(d,J C-P =122Hz),132.2(d,J C-P =3Hz),132.1(d,J C-P =3Hz),131.5(d,J C-P =10Hz),131.4(d,J C-P =10Hz),128.7(d,J C-P =12Hz),128.6(d,J C-P =12Hz),126.2(d,J C-F =9Hz),114.0(d,J C-F =22Hz),110.9(d,J C-F =23Hz),103.3(dd,J C-P =114Hz,J C-P =3Hz),37.8(d,J C-P =12Hz); 31 P NMR(162MHz,CDCl3):δ31.3,18.3; 19 F NMR(376MHz,CDCl3):δ-113.9;ESI-HRMS:m / z Calcd for C 33 H 26 FNO2P2:549.1423,found 549.1419。

[0058] Synthesis of Compound 13: 5-Chloro-1-indanone oxime (73.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 13 (176 mg, 78% yield) as a white solid. Product monitoring data are as follows:

[0059] 1 H NMR (400MHz, CDCl3): δ10.06(d,J=10.0Hz,1H),8.60(d,J=8.4Hz,1H),7.99(dd,J=12.4Hz,J=6.8Hz,4H),7.71(dd,J=12.4H z,J=6.8Hz,4H),7.61-7.57(m,2H),7.53-7.49(m,4H),7.46-7.36(m,6H),7.22(s,1H),7.20(d,J=7.6Hz,1H),3.25(s,2H); 13 CNMR (100MHz, CDCl3): δ156.9(d,J C-P =6Hz),146.5(d,J C-P =12Hz),136.9(d,J C-P =15Hz),133.8,132.6(d,J C-P =118Hz),132.5(d,J C-P =116Hz),132.2(d,J C-P =2Hz),132.0(d,J C-P =2Hz),131.5(d,J C-P =11Hz),131.4(d,J C-P =10Hz),128.7(d,J C-P =12Hz),128.6(d,J C-P =13Hz),127.1,125.8,123.8(d,J C-P =2Hz),104.1(dd,J C-P =112Hz,J C-P=8Hz),37.7(d,J C-P =12Hz); 31 P NMR (162MHz, CDCl3): δ31.4,18.3; ESI-HRMS: m / z Calcd for C 33 H 26 ClNO2P2:565.1127, found 565.1122.

[0060] Synthesis of Compound 14: 5-Bromo-1-indanone oxime (91.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed. The product 14 (129 mg, 53% yield) was obtained as a white solid. Product monitoring data are as follows:

[0061] 1 H NMR (400MHz, CDCl3): δ10.05(d,J=9.6Hz,1H),8.55(d,J=8.8Hz,1H),7.98(dd,J=12.4Hz,J=6.8Hz,4H),7.71(dd,J=1 2.4Hz,J=6.8Hz,4H),7.59(t,J=7.2Hz,2H),7.53-7.49(m,4H),7.44(t,J=7.2Hz,2H),7.46-7.36(m,6H),3.24(s,2H); 13 C NMR (100MHz, CDCl3): δ157.0(d,J C-P =4Hz),146.8(d,J C-P =11Hz),137.4(d,J C-P =15Hz),132.6(d,J C-P =119Hz),132.5(d,J C-P =114Hz),132.3(d,J C-P =2Hz),132.0(d,J C-P =2Hz),131.6(d,J C-P=11Hz),131.5(d,J C-P =10Hz),130.0,128.8(d,J C-P =12Hz),128.7(d,J C-P =13Hz),126.8(d,J C-P =2Hz),126.2,122.2,104.1(dd,J C-P =112Hz,J C-P =7Hz),37.8(d,J C-P =12Hz); 31 P NMR (162MHz, CDCl3): δ31.2,18.2; ESI-HRMS: m / z Calcd for C 33 H 26 BrNO2P2:609.0622,found609.0615.

[0062] Synthesis of Compound 15: 5-iodo-1-indanone oxime (109.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 15 (200 mg, 76% yield) as a white solid. Product monitoring data are as follows:

[0063] 1 H NMR (400MHz, CDCl3): δ10.05(d,J=10.0Hz,1H),8.41(d,J=8.4Hz,1H),7.98(dd,J=12.4Hz,J=6.8Hz,4H),7.71(dd, J=12.4Hz, J=6.8Hz,4H),7.61-7.57(m,3H),7.54-7.49(m,5H),7.46-7.44(m,2H),7.42-7.36(m,4H),3.22(s,2H); 13 C NMR (100MHz, CDCl3): δ157.1(d,J C-P =6Hz),146.9(d,J C-P=12Hz),138.0(d,J C-P =15Hz),135.8,132.7,132.5(d,J C-P =121Hz),132.4(d,J C-P =113Hz),132.3(d,J C-P =2Hz),132.0(d,J C-P =2Hz),131.5(d,J C-P =10Hz),131.4(d,J C-P =10Hz),128.6(d,J C-P =13Hz),128.5(d,J C-P =13Hz),126.3,103.9(dd,J C-P =112Hz,J C-P =7Hz),94.1,37.6(d,J C-P =13Hz); 31 P NMR (162MHz, CDCl3): δ31.3,18.3; ESI-HRMS: m / z Calcd for C 33 H 26 INO2P2:657.0483,found 657.0478.

[0064] Synthesis of Compound 16: 5-Methyl-1-indanone oxime (65.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 16 (118 mg, 54% yield) as a white solid. Product monitoring data are as follows:

[0065] 1H NMR(400MHz,CDCl3):δ10.03(d,J=10.0Hz,1H),8.47(d,J=8.0Hz,1H),7.99(dd,J=12.4Hz,J=6.8Hz,4H),7.72(dd,J=12.4Hz,J=6.8Hz,4H),7.57(t,J=6.8Hz,2H),7.52-7.47(m,4H),7.43-7.35(m,6H),7.05(s,1H),7.03(d,J=9.2Hz,1H),3.23(s,2H),2.29(s,3H); 13 C NMR(100MHz,CDCl3):δ157.8(d,J C-P =6Hz),145.3(d,J C-P =11Hz),137.6,133.5,132.0(d,J C-P =3Hz),131.8(d,J C-P =3Hz),131.7(d,J C-P =147Hz),131.61(d,J C-P =136Hz),131.6(d,J C-P =10Hz),131.5(d,J C-P =10Hz),128.6(d,J C-P =12Hz),128.5(d,J C-P =12Hz),127.7,124.5,124.2(d,J C-P =2Hz),102.2(dd,J C-P =114Hz,J C-P =7Hz),37.7(d,J C-P =12Hz),21.4; 31 P NMR(162MHz,CDCl3):δ31.7,17.9;ESI-HRMS:m / z Calcd forC 34 H 29 NO2P2:545.1674,found 545.1671。

[0066] Synthesis of Compound 17: 5-Methoxy-1-indanone oxime (71.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 17 (72 mg, 32% yield) as a yellow solid. Product monitoring data are as follows:

[0067] 1 H NMR (400MHz, CDCl3): δ10.04 (d, J = 10.0Hz, 1H), 8.51 (d, J = 8.4Hz, 1H), 7.99 (dd, J = 12.4Hz, J = 6.8Hz, 4H), 7.72 (dd, J = 12. 4Hz,J=6.8Hz,4H),7.59-7.56(m,2H),7.52-7.48(m,4H),7.46-7.37(m,6H),6.80-6.76(m,2H),3.74(s,3H),3.24(s,2H); 13 C NMR (100MHz, CDCl3): δ159.7,157.6(d,J C-P =5Hz),147.3(d,J C-P =12Hz),132.9(d,J C-P =139Hz),132.8(d,J C-P =147Hz),132.6,132.0(d,J C-P =3Hz),131.9(d,J C-P =3Hz),131.6(d,J C-P =11Hz),131.5(d,J C-P =10Hz),128.7(d,J C-P =13Hz),128.6(d,J C-P =12Hz),125.8,112.4,109.5(d,J C-P =3Hz),100.7(dd,J C-P =115Hz,J C-P =7Hz),55.3,37.8(d,JC-P =12Hz); 31 P NMR (162MHz, CDCl3): δ31.6,18.0; ESI-HRMS: m / z Calcd for C 34 H 29 NO3P2:561.1623, found 561.1619.

[0068] Synthesis of Compound 18: 6-Methoxy-1-indanone oxime (71.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 18 (197 mg, 88% yield) as a yellow solid. Product monitoring data are as follows:

[0069] 1 H NMR (400MHz, CDCl3): δ10.05(d,J=10.0Hz,1H),8.32(d,J=2.0Hz,1H),8.02(dd,J=12.4Hz,J=6.8Hz,4H),7.73(dd,J=12.4Hz,J=6.8Hz,4H),7 .58(t,J=7.6Hz,2H),7.52-7.48(m,4H),7.46-7.37(m,6H),7.10(d,J=8.0Hz,1H),6.77(dd,J=8.0Hz,J=2.0Hz,1H),3.79(s,3H),3.22(s,2H); 13 C NMR (100MHz, CDCl3): δ158.8,157.6(d,J C-P =8Hz),139.5(d,J C-P =12Hz),137.1(d,J C-P =12Hz),132.74(d,J C-P =127Hz),132.72(d,J C-P =127Hz),132.1(d,J C-P =2Hz),132.0(d,J C-P=3Hz),131.5(d,J C-P =10Hz),131.4(d,J C-P =10Hz),128.7(d,J C-P =13Hz),128.6(d,J C-P =13Hz),123.9,116.1,107.8,104.7(dd,J C-P =113Hz,J C-P =7Hz),55.5,37.3(d,J C-P =12Hz); 31 P NMR (162MHz, CDCl3): δ31.5,18.2; ESI-HRMS: m / zCalcd for C 34 H 29 NO3P2:561.1623, found 561.1620.

[0070] Synthesis of Compound 19: 4-Methoxy-1-indanone oxime (71.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 19 (204 mg, 91% yield) as a yellow solid. Product monitoring data are as follows:

[0071] 1 H NMR (400MHz, CDCl3): δ10.06(d,J=9.6Hz,1H),8.25(d,J=8.0Hz,1H),8.00(dd,J=12.4Hz,J=6.8Hz,4H),7.73(dd,J=12.4Hz,J=6.8Hz,4 H),7.56(t,J=7.6Hz,2H),7.51-7.47(m,4H),7.44-7.35(m,6H),7.21(t,J=8.0Hz,1H),6.72(d,J=8.0Hz,1H),3.75(s,3H),3.23(s,2H); 13 C NMR (100MHz, CDCl3): δ157.5(d,JC-P =6Hz),154.4,140.0(d,J C-P =13Hz),132.9(d,J C-P =119Hz),132.7(d,J C-P =120Hz),132.3,132.2,132.0(d,J C-P =3Hz),131.8(d,J C-P =3Hz),131.5(d,J C-P =10Hz),131.4(d,J C-P =10Hz),128.5(d,J C-P =12Hz),128.4(d,J C-P =9Hz),117.6,116.1,109.3,103.7(dd,J C-P =113Hz,J C-P =7Hz),55.0,35.1(d,J C-P =13Hz); 31 P NMR (162MHz, CDCl3): δ31.6,17.8; ESI-HRMS: m / z Calcd forC 34 H 29 NO3P2:561.1623, found 561.1621.

[0072] Synthesis of Compound 20: 5,6-dimethoxy-1-indanone oxime (83.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 20 (73 mg, 31% yield) as a yellow solid. Product monitoring data are as follows:

[0073] 1H NMR(600MHz,CDCl3):δ10.03(d,J=10.2Hz,1H),8.33(s,1H),8.01(dd,J=12.6Hz,J=7.2Hz,4H),7.73(dd,J=12.6Hz,J=7.2Hz,4H),7.58(t,J=7.2Hz,2H),7.52-7.49(m,4H),7.44(t,J=7.2Hz,2H),7.41-7.38(m,4H),6.79(s,1H),3.88(s,1H),3.80(s,3H),3.23(s,2H); 13 C NMR(150MHz,CDCl3):δ157.8(d,J C-P =4Hz),149.1,148.0,138.4(d,J C-P =10Hz),133.6,132.7(d,J C-P =128Hz),132.4(d,J C-P =128Hz),132.0,132.0,131.5(d,J C-P =10Hz),131.4(d,J C-P =10Hz),128.6(d,J C-P =16Hz),128.5(d,J C-P =16Hz),107.6,106.4,101.4(d,J C-P =110Hz),56.1,55.8,37.8(d,J C-P =12Hz); 31 P NMR(162MHz,CDCl3):δ31.4,18.2;ESI-HRMS:m / zCalcd for C 35 H 31 NO4P2:591.1728,found 591.1728。

[0074] Synthesis of Compound 21: 3-Benzofuranoside oxime (60.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 21 (188 mg, 88% yield) as a white solid. Product monitoring data are as follows:

[0075] 1 H NMR (400MHz, CDCl3): δ8.64 (d, J = 10.8Hz, 1H), 8.49 (d, J = 8.0Hz, 1H), 8.00 (dd, J = 12.4Hz, J = 6.8Hz, 4H), 7.80 (dd, J = 12.4Hz, J = 6.8 Hz,4H),7.59(t,J=7.6Hz,2H),7.52-7.48(m,4H),7.43(t,J=7.6Hz,2H),7.38-7.34(m,4H),7.28-7.25(m,2H),7.17-7.13(m,1H); 13 C NMR (100MHz, CDCl3): δ156.6(d,J C-P =9Hz),138.6(d,J C-P =15Hz),132.6(d,J C-P =3Hz),132.3(d,J C-P =126Hz),132.0(d,J C-P =3Hz),131.6(d,J C-P =109Hz),131.6(d,J C-P =9Hz),131.5(d,J C-P =11Hz),128.7(d,J C-P =12Hz),128.7(d,J C-P =12Hz),128.3(dd,J C-P =138Hz,J C-P =8Hz),127.3,124.5,122.9,121.6(d,J C-P =10Hz),111.5;31 P NMR (162MHz, CDCl3): δ24.9,18.6; ESI-HRMS: m / z Calcd for C 32 H 25 NO3P2:533.1310,found533.1309.

[0076] Synthesis of Compound 22: 6-Fluoro-3-benzofuranone oxime (67.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 22 (117 mg, 53% yield) as a white solid. Product monitoring data are as follows:

[0077] 1 H NMR (400MHz, CDCl3): δ8.64 (d, J=10.8Hz, 1H), 8.50 (dd, J=8.8Hz, J=5.2Hz, 1H), 7.99 (dd, J=12.4Hz, J=7.2Hz, 4H), 7.78 (dd, J=12.4Hz, J=7.2Hz, 4H), 7.60(t,J=7.2Hz,2H),7.53-7.49(m,4H),7.45(t,J=7.2Hz,2H),7.39-7.35 (m,4H),6.98(dd,J=8.8Hz,J=2.0Hz,1H),6.92(dt,J=5.2Hz,J=2.0Hz,1H); 13 C NMR (100MHz, CDCl3): δ162.5(d,J C-F =246Hz),156.8(d,J C-P =13Hz,J C-F =9Hz),138.5(d,J C-F =14Hz),132.7(d,J C-P =3Hz),132.12(d,J C-P =3Hz),132.1(d,J C-P =126Hz),131.5(d,JC-P =110Hz),131.5(d,J C-P =10Hz),131.4(d,J C-P =11Hz),129.1(J C-P =138Hz),128.8(d,J C-P =13Hz),128.7(d,J C-P =13Hz),125.6(d,J C-F =11Hz),118.2(d,J C-F =8Hz),111.7(d,J C-F =24Hz),99.0(d,J C-F =26Hz); 31 P NMR (162MHz, CDCl3): δ24.7,19.0; 19 F NMR (376MHz, CDCl3): δ-112.5; ESI-HRMS: m / z Calcd for C 32 H 24 FNO3P2:551.1215, found 551.1212.

[0078] Synthesis of Compound 23: 5-Methoxy-3-benzofuranone oxime (72.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 23 as a white solid (115 mg, 51% yield). Product monitoring data are as follows:

[0079] 1H NMR(400MHz,CDCl3):δ8.60(d,J=10.8Hz,1H),8.01(dd,J=12.4Hz,J=6.8Hz,4H),8.00(d,J=8.4Hz,1H),7.79(dd,J=12.8Hz,J=6.8Hz,4H),7.59(t,J=7.6Hz,2H),7.52-7.48(m,4H),7.44(t,J=7.6Hz,2H),7.40-7.35(m,4H),7.16(d,J=9.2Hz,1H),6.88(dd,J=9.2Hz,J=2.4Hz,1H),3.78(s,3H); 13 C NMR(100MHz,CDCl3):δ155.6,151.7(d,J C-P =8Hz),138.5(d,J C-P =15Hz),132.6(d,J C-P =3Hz),132.3(d,J C-P =127Hz),132.1(d,J C-P =3Hz),131.6(d,J C-P =110Hz),131.5(d,J C-P =10Hz),131.4(d,J C-P =11Hz),129.1(dd,J C-P =138Hz,J C-P =8Hz),128.8(d,J C-P =13Hz),128.7(d,J C-P =13Hz),121.8(d,J C-P =6Hz),117.9,112.2,104.5,55.7; 31 P NMR(162MHz,CDCl3):δ24.8,18.6;ESI-HRMS:m / z Calcd forC 33 H 27 NO4P2:563.1415,found 563.1412。

[0080] Synthesis of Compound 24: 7-Methoxy-3-benzofuranone oxime (72.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 24 (160 mg, 71% yield) as a white solid. Product monitoring data are as follows:

[0081] 1 H NMR (400MHz, CDCl3): δ8.67(d,J=10.8Hz,1H),8.07(d,J=8.4Hz,1H),8.00(dd,J=12.4Hz,J=7.2Hz,4H),7.81(dd,J=13.2Hz,J=7.2Hz,4H),7 .57(t,J=7.6Hz,2H),7.51-7.47(m,4H),7.42(t,J=7.2Hz,2H),7.38-7.33(m,4H),7.06(d,J=8.0Hz,1H),6.75(d,J=8.0Hz,1H),3.83(s,3H); 13 C NMR (100MHz, CDCl3): δ146.5(d,J C-P =9Hz),145.2,139.0(d,J C-P =15Hz),132.5(d,J C-P =2Hz),132.3(d,J C-P =127Hz),131.9(d,J C-P =3Hz),131.6(d,J C-P =110Hz),131.5(d,J C-P =10Hz),131.4(d,J C-P =11Hz),130.6(d,J C-P =10Hz),128.7(d,J C-P =13Hz),128.6(d,J C-P =12Hz),128.5(dd,J C-P =137Hz,J C-P=8Hz),123.4,116.2,108.8,55.7; 31 P NMR (162MHz, CDCl3): δ25.0,18.4; ESI-HRMS: m / z Calcd forC 33 H 27 NO4P2:563.1415, found 563.1414.

[0082] Synthesis of Compound 25: To a 38 mL oven-dried reaction tube were added isochroman-4-one oxime (66.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL). The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 25 as a white solid (112 mg, 51% yield). Product monitoring data are as follows:

[0083] 1 H NMR (400MHz, CDCl3): δ8.68(d,J=8.4Hz,1H),8.30(d,J=8.0Hz,1H),7.88(dd,J=12.0Hz,J=2.8Hz,4H),7.77(dd,J=12.4Hz,J=3 .2Hz,4H),7.58(t,J=7.2Hz,2H),7.50-7.46(m,5H),7.42-7.36(m,3H),7.27-7.22(m,4H),6.89(d,J=7.2Hz,1H),4.48(s,2H); 13 C NMR (100MHz, CDCl3): δ138.5(d,J C-P =14Hz),132.3(d,J C-P =3Hz),131.8(d,J C-P =124Hz),131.7(d,J C-P =10Hz),131.6(d,J C-P =11Hz),131.5(d,J C-P =3Hz),131.0(d,J C-P =107Hz),130.7(d,JC-P =12Hz),130.6(d,J C-P =9Hz),129.5,129.1(d,J C-P =125Hz),128.9(d,J C-P =13Hz),128.5(d,J C-P =13Hz),128.1(d,J C-P =13Hz),126.4,123.3,68.5(d,J C-P =5Hz); 31 P NMR (162MHz, CDCl3): δ27.8,20.8; ESI-HRMS: m / z Calcd for C 33 H 27 NO3P2:547.1466, found 547.1462.

[0084] Synthesis of Compound 26: 8-Bromo-3,4-dihydro-2H-benzo[B]oxepin-5-one oxime (103.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed. Finally, a white solid product 26 (120 mg, 47% yield) was obtained. The product monitoring data are as follows:

[0085] 1 H NMR (400MHz, CDCl3): δ10.36(d,J=10.4Hz,1H),7.98(d,J=8.4Hz,1H),7.84-7.76(m,8H),7.62(t,J=7.2H z,2H),7.56-7.49(m,4H),7.39-7.32(m,7H),6.77(d,J=2.0Hz,1H),3.82-3.77(m,2H),2.00-1.90(m,2H); 13 C NMR (100MHz, CDCl3): δ155.5,154.9(d,J C-P =3Hz),132.4(d,J C-P=3Hz),132.4(d,J C-P =3Hz),132.2(d,J C-P =131Hz),132.1(d,J C-P =135Hz),131.8(d,J C-P =10Hz),131.6,131.4(d,J C-P =10Hz),128.8(d,J C-P =12Hz),128.8(d,J C-P =12Hz),128.2(d,J C-P =13Hz),126.9,124.7,124.3,99.1(dd,J C-P =101Hz,J C-P =6Hz),79.3,27.4(d,J C-P =12Hz); 31 P NMR (162MHz, CDCl3): δ37.8,16.4; ESI-HRMS: m / zCalcd for C 34 H 28 BrNO3P2:639.0728, found 639.0723.

[0086] Synthesis of Compound 27: To a 38 mL oven-dried reaction tube were added diphenylacetone oxime (85.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL). The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 27 (117 mg, 49% yield) as a white solid. Product monitoring data are as follows:

[0087] 1H NMR(400MHz,CDCl3):δ11.06(d,J=11.6Hz,1H),7.65(dd,J=12.4Hz,J=7.2Hz,4H),7.55(dd,J=12.0Hz,J=7.2Hz,4H),7.44(t,J=7.2Hz,2H),7.35-7.29(m,6H),7.26-7.19(m,4H),6.87-6.79(m,3H),6.71(t,J=7.6Hz,2H),6.64(t,J=7.6Hz,2H),6.42(d,J=7.2Hz,2H),4.48(s,2H); 13 C NMR(150MHz,CDCl3):δ158.7(d,J C-P =3Hz),135.5(d,J C-P =9Hz),134.8(d,J C-P =9Hz),133.2(d,J C-P =129Hz),132.8(d,J C-P =3Hz),132.2(d,J C-P =9Hz),131.8(d,J C-P =2Hz),131.6(d,J C-P =104Hz),131.4(d,J C-P =11Hz),131.3(d,J C-P =2Hz),130.6,128.2(d,J C-P =14Hz),128.1(d,J C-P =12Hz),127.9,127.3,126.5,126.1,106.3(dd,J C-P =98Hz,J C-P =6Hz); 31 P NMR(162MHz,CDCl3):δ36.0,16.0;ESI-HRMS:m / zCalcd for C 38 H 31 NO2P2:595.1830,found 595.1826。

[0088] Synthesis of Compound 28: 4'-Methoxy-2-phenylacetophenone oxime (97.0 mg, 0.40 mmol, 1.0 equiv), diphenylphosphine oxide (324 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 28 (145 mg, 58% yield) as a white solid. Product monitoring data are as follows:

[0089] 1 H NMR (400MHz, CDCl3): δ11.02(d,J=11.6Hz,1H),7.73(dd,J=12.4Hz,J=7.2Hz,4H),7.62(dd,J=12.0Hz,J=7.2Hz,4H),7.50(t,J=7.6Hz,2H),7.42-7.37( m,6H),7.34-7.30(m,4H),6.87(d,J=8.8Hz,2H),6.78(d,J=7.6Hz,1H),6.74 (t,J=8.0Hz,2H),6.51(d,J=6.8Hz,2H),6.32(d,J=8.8Hz,2H),3.62(s,3H); 13 C NMR (150MHz, CDCl3): δ159.0,158.7(d,J C-P =4Hz),135.8(d,J C-P =10Hz),133.8,132.5,132.3(d,J C-P =95Hz),132.26(d,J C-P =94Hz),132.24(d,J C-P =13Hz),132.2(d,J C-P =3Hz),131.4(d,J C-P =14Hz),131.3(d,J C-P =4Hz),128.2(d,J C-P =13Hz),128.1(d,J C-P =12Hz),127.4,127.3,126.0(d,J C-P=3Hz),112.0,106.1(dd,J C-P =97Hz,J C-P =7Hz),54.9; 31 P NMR (162MHz, CDCl3): δ36.1,16.0; ESI-HRMS: m / zCalcd for C 39 H 33 NO3P2:625.1936,found 625.1931.

[0090] Synthesis of Compound 29: 1-Indanone oxime (59.0 mg, 0.40 mmol, 1.0 equiv), di-n-butylphosphine oxide (260 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in a 120°C oil bath for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography was performed (petroleum ether / ethyl acetate = 1:1 to 0:1). The product 29 (114 mg, 63% yield) was obtained as a white solid. Product monitoring data are as follows:

[0091] 1 H NMR (400MHz, CDCl3): δ9.31 (d, J = 8.4Hz, 1H), 8.67 (d, J = 8.0Hz, 1H), 7.42 (t, J = 7.2Hz, 1H), 7.40 (d, J = 8.0Hz, 1H), 7.33(t,J=7.2Hz,1H),3.28(s,2H),1.96-1.83(m,6H),1.68-1.60(m,8H),1.44-1.37(m,10H),0.92-0.87(m,12H); 13 C NMR (100MHz, CDCl3): δ157.9,144.4(d,J C-P =10Hz),138.7(d,J C-P =12Hz),127.3,127.1,124.2,123.7(d,J C-P =2Hz),101.3(d,J C-P =108Hz),36.3(d,J C-P =12Hz),30.1(d,J C-P=117Hz),29.3(d,J C-P =130Hz),24.2(d,J C-P =4Hz),24.0(d,J C-P =14Hz),23.9(d,J C-P =15Hz),23.4(d,J C-P =4Hz),13.60,13.55; 31 P NMR (162MHz, CDCl3): δ47.3, 40.4; ESI-HRMS: m / z Calcd for C 25 H 43 NO2P2:451.2769, found 451.2767.

[0092] Synthesis of Compound 30: 1-indanone oxime (59.0 mg, 0.40 mmol, 1.0 equiv), bis(4-methoxyphenyl)phosphine oxide (420 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1 to 0:1) was performed to obtain the product 30 (198 mg, 76% yield) as a white solid. The product monitoring data are as follows:

[0093] 1 H NMR (400MHz, CDCl3): δ9.91(d,J=9.6Hz,1H),8.61(d,J=8.4Hz,1H),7.91(dd,J=11.6Hz,J=8.8Hz,4H),7.64(dd,J=11.6Hz,J=8.8Hz,4H),7.25 -7.21(m,2H),7.17(t,J=7.2Hz,1H),6.99(dd,J=8.8Hz,J=2.0Hz,4H),6.88(dd,J=8.8Hz,J=2.4Hz,4H),3.84(s,6H),3.75(s,6H),3.24(s,2H); 13 C NMR (100MHz, CDCl3): δ162.4(d,J C-P =3Hz),162.1(d,J C-P=3Hz),157.0(d,J C-P =6Hz),144.7(d,J C-P =11Hz),138.6(d,J C-P =13Hz),133.4(d,J C-P =14Hz),133.3(d,J C-P =13Hz),127.1,126.6,124.8,124.6(d,J C-P =134Hz),124.4(d,J C-P =111Hz),123.3(d,J C-P =1Hz),114.0(d,J C-P =13Hz),114.0(d,J C-P =13Hz),104.4(dd,J C-P =115Hz,J C-P =8Hz),55.3,55.1,37.9(d,J C-P =12Hz); 31 P NMR (162MHz, CDCl3): δ30.7,18.2; ESI-HRMS: m / z Calcd for C 37 H 35 NO6P2:651.1940, found 651.1938.

[0094] Synthesis of Compound 31: 1-Indanone oxime (59.0 mg, 0.40 mmol, 1.0 equiv), bis(p-methylphenyl)phosphine oxide (369 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 31 (138 mg, 59% yield) as a white solid. Product monitoring data are as follows:

[0095] 1H NMR(400MHz,CDCl3):δ10.00(d,J=10.0Hz,1H),8.60(d,J=8.8Hz,1H),7.87(dd,J=12.4Hz,J=8.4Hz,4H),7.60(dd,J=12.0Hz,J=8.0Hz,4H),7.29(dd,J=8.0Hz,J=2.0Hz,4H),7.24-7.14(m,7H),3.24(s,2H),2.42(s,6H),2.30(s,6H); 13 C NMR(100MHz,CDCl3):δ157.3(d,J C-P =6Hz),144.9(d,J C-P =12Hz),142.4(d,J C-P =3Hz),142.1(d,J C-P =3Hz),138.5(d,J C-P =11Hz),131.6(d,J C-P =11Hz),131.5(d,J C-P =11Hz),130.0(d,J C-P =120Hz),129.8(d,J C-P =119Hz),129.3(d,J C-P =12Hz),129.2(d,J C-P =13Hz),127.1,126.7,124.8,123.3(d,J C-P =2Hz),103.8(dd,J C-P =114Hz,J C-P =8Hz),37.9(d,J C-P =12Hz),21.6,21.5; 31 P NMR(162MHz,CDCl3):δ31.5,18.5;ESI-HRMS:m / z Calcd for C 37 H 35 NO2P2:587.2143,found 587.2143。

[0096] Synthesis of Compound 32: 1-Indanone oxime (59.0 mg, 0.40 mmol, 1.0 equiv), bis(3,5-dimethylphenyl)phosphine oxide (414 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was then stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 1:1) was performed to obtain the product 32 (167 mg, 65% yield) as a white solid. Product monitoring data are as follows:

[0097] 1 H NMR (400MHz, CDCl3): δ9.92(d,J=9.6Hz,1H),8.56(d,J=7.6Hz,1H),7.56(d,J=12.8Hz,4H),7.32(d,J=12.4Hz,4H) ,7.26(t,J=7.2Hz,2H),7.20(t,J=7.2Hz,1H),7.17(s,2H),7.03(s,2H),3.29(s,2H),2.34(s,12H),2.24(s,12H); 13 C NMR (100MHz, CDCl3): δ157.4(d,J C-P =8Hz),145.0(d,J C-P =11Hz),138.8(d,J C-P =12Hz),138.2(d,J C-P =13Hz),138.0(d,J C-P =14Hz),133.6(d,J C-P =3Hz),133.5(d,J C-P =4Hz),133.1(d,J C-P =117Hz),132.8(d,J C-P =126Hz),129.2(d,J C-P =10Hz),129.0(d,J C-P =11Hz),127.0,126.7,124.9,123.2(d,J C-P =2Hz),103.9(dd,J C-P =112Hz,J C-P=7Hz),38.2(d,J C-P =12Hz),21.3,21.2; 31 P NMR (162MHz, CDCl3): δ31.4,19.6; ESI-HRMS: m / z Calcd for C 41 H 43 NO2P2:643.2769, found 643.2771.

[0098] Synthesis of compound 33: 1-indanone oxime (59.0 mg, 0.40 mmol, 1.0 equiv), bis(3,5-di-tert-butylphenyl)phosphine oxide (683 mg, 1.60 mmol, 4.0 equiv), DTBP (0.22 mL, 1.20 mmol, 3.0 equiv), acetic acid (70 μL, 1.20 mmol, 3.0 equiv), and DMF (2 mL) were added sequentially to a 38 mL oven-dried reaction tube. The reaction tube was evacuated, purged with nitrogen, and sealed. The mixture was stirred in an oil bath at 120°C for approximately 30 hours. After the starting material was consumed, the system was cooled to room temperature, and the resulting residue was extracted with ethyl acetate (15 mL x 5). The organic phases were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate = 10:1) was performed to obtain product 33 as a white solid (274 mg, 70% yield). The product monitoring data are as follows:

[0099] 1 H NMR (400MHz, CDCl3): δ10.26(d,J=10.0Hz,1H),8.53(d,J=5.6Hz,1H),7.89(dd,J=13.2Hz,J=1.6Hz,4H),7.56(s,2H),7.51(dd,J=1 3.2Hz,J=1.6Hz,4H),7.44(s,2H),7.21(d,J=3.2Hz,1H),7.12(dd,J=5.6Hz,J=3.2Hz,2H),3.30(s,2H),1.28(s,36H),1.23(s,36H); 13 C NMR (100MHz, CDCl3): δ157.5(d,J C-P =6Hz),150.7(d,J C-P =11Hz),150.6(d,J C-P =13Hz),144.7(d,J C-P =10Hz),138.6(d,J C-P =12Hz),132.8(d,J C-P =104Hz),132.2(d,JC-P =126Hz),126.7,126.3(d,J C-P =11Hz),126.2(d,J C-P =3Hz),125.7(d,J C-P =12Hz),125.6(d,J C-P =11Hz),125.5(d,J C-P =3Hz),125.0,122.9,103.4(dd,J C-P =111Hz,J C-P =7Hz),38.1(d,J C-P =12Hz),35.0,34.9,31.4,31.3; 31 P NMR (162MHz, CDCl3): δ32.2, 20.0; ESI-HRMS: m / z Calcd for C 65 H 91 NO2P2:979.6525, found 979.6515.

[0100] The above compounds can be prepared with reference to the literature (Inorg. Synth., 1989, 25, 129; J. Chem. Soc., Dalton Trans., 2001, 15, 2279; J. Organomet. Chem., 1999, 582, 83).

[0101] Example 2 Effects of IDPA and its homologues on PRRSV replication in MARC-145 cells

[0102] 1. Configuration of IDPA and its homologues

[0103] 5 mg of dry powder of IDPA (compound 2) and its homologues (compounds 3-33) were weighed respectively, fully dissolved in the corresponding volume of DMSO to prepare a 20 mM mother solution, filtered through a 0.22 μm filter and stored in a -80°C refrigerator.

[0104] 2. Cell Inoculation and Use of IDPA and Its Homologs

[0105] Cell plating: MARC-145 cells were plated at 1×10 5 Cells were seeded at a density of 100 cells / well in a 12-well cell plate and cultured in a cell culture incubator at 37°C containing 5% CO2.

[0106] Inoculation: When the cell confluence reaches about 80%, inoculate 1 MOI of PRRSV SD-YL1712 strain. According to the formula PFU = number of cells × MOI = 0.7 × TCID 50Calculate the required volume of virus solution. Dilute the required volume of virus solution to 1 mL of fresh serum-free DMEM medium. Remove the 12-well plate, discard the medium, inoculate the diluted virus solution into the 12-well plate, and incubate in a 37°C cell culture incubator with 5% CO2 for 1 hour.

[0107] IDPA treatment: Remove the 12-well plate, discard the virus solution, take 0.5 μL of IDPA homologue stock solution and add it to 1 mL of DMEM medium containing 3% serum, mix well and add it to the 12-well plate; the control group is treated with DMSO;

[0108] Sampling: Cell samples and cell culture supernatant were collected 24 h and 36 h after infection and stored at -80°C. Cell samples were used to detect the expression level of PRRSV ORF7 gene and PRRSV-N protein in cells, and cell culture supernatant was used to detect the titer of virus released into the culture supernatant.

[0109] 3. RT-qPCR detection of PRRSV ORF7 gene expression level in MARC-145 cells

[0110] Extraction of total RNA: Add 400 μL TRIzol lysis buffer to the collected cell sample, vortex to lyse the cells, let stand for 10 minutes, then add 100 μL chloroform, vortex to mix, let stand for 10 minutes, and then centrifuge at 13,000 rpm in a 4°C centrifuge for 10 minutes. Transfer the supernatant to a new 1.5 mL sterile, enzyme-free centrifuge tube, add an equal volume of isopropanol, vortex to mix, let stand for 10 minutes, and then centrifuge at 13,000 rpm in a 4°C centrifuge for 10 minutes. Discard the liquid, add 75% ethanol, invert to mix, and centrifuge at 13,000 rpm in a 4°C centrifuge for 5 minutes. Discard the liquid and air dry on the workbench. Add ddH2O to dissolve the RNA for later use.

[0111] Reverse transcription: Determine the total RNA concentration in the sample, then use a reverse transcription kit to obtain cDNA of the sample, which is then tested by RT-qPCR;

[0112] RT-qPCR detection: The PRRSV ORF7 gene and the internal reference gene GAPDH in the samples were detected using an RT-qPCR detection kit, and the data were analyzed using GraphPad Prism8.

[0113] like Figure 1 As shown, compared with the DMSO control group, the expression level of PRRSV ORF7 gene was reduced when MARC-145 cells were infected with PRRSV SD-YL1712 strain and treated with IDPA for 24h and 36h.

[0114] like Figure 2As shown, compared with the DMSO control group, after MARC-145 cells were infected with PRRSV SD-YL1712 strain and treated with different doses (2, 4, 6, 8, 10 μM) of IDPA for 24 h, the expression level of PRRSV ORF7 gene was reduced in a dose-dependent manner.

[0115] like Figure 3 As shown, compared with the DMSO control group, after PRRSV SD-YL1712 strain infected MARC-145 cells, the expression level of PRRSV ORF7 gene was reduced when the cells were treated with 10 μM IDPA homologs 4, 6, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33 for 24 hours; and after the cells were treated with 10 μM IDPA homologs 3, 5, 7 for 24 hours, the expression level of PRRSV ORF7 gene was reduced. There was no significant difference in the expression level of ORF7 gene; these results indicate that IDPA homologs 4, 6, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, and 33 have significant inhibitory effects on PRRSV replication, while IDPA homologs 3, 5, and 7 have no significant inhibitory effects on PRRSV replication.

[0116] 4. Western blot detection of PRRSV-N protein expression level in MARC-145 cells

[0117] Cell lysis: Add 120 μL of high-efficiency RIPA tissue / cell rapid lysis buffer (containing protease inhibitors, 1:100) to the collected cell sample, repeatedly pipette to dissolve the cells, and then place on ice for 20-30 minutes. After lysis is complete, centrifuge at 13,000 rpm in a 4°C centrifuge for 10 minutes. Transfer the supernatant to a new 1.5 mL sterile, enzyme-free centrifuge tube.

[0118] Protein quantification: BCA protein quantification kit (Shanghai Biotech Co., Ltd.) was used for protein quantification according to the manufacturer's instructions. The protein content in each sample was standardized to 3 μg / μL based on the protein quantification results.

[0119] Sample preparation: Add 20 μL of prepared 5× SDS Loading Buffer to 80 μL of cell lysis supernatant, mix thoroughly, and boil in a water bath for 10 min. Remove the sample and proceed to SDS-PAGE electrophoresis.

[0120] SDS-PAGE electrophoresis: Prepare 12% SDS-PAGE gel as required, load samples in order, run the stacking gel at 90V first, then adjust the voltage to 200V and continue electrophoresis;

[0121] Transfer: After SDS-PAGE electrophoresis, remove the gel and place it in transfer buffer. Immerse the cut PVDF membrane in methanol for 60 seconds to fully activate it. Fasten the transfer clamps in the order of clamps, sponge, filter paper, gel, PVDF membrane, filter paper, sponge, and clamps. Remove all air bubbles between the membrane and gel to ensure a smooth transfer. Transfer at a constant voltage of 100V for 70-90 minutes, depending on the size of the protein to be tested.

[0122] Blocking: After transfer, place the PVDF membrane in the prepared 5% skim milk powder blocking solution and block at room temperature for 2 hours;

[0123] Incubate with primary antibodies: Dilute the desired primary antibodies, PRRSV-N protein monoclonal antibody (1:2000) and β-actin monoclonal antibody (1:2000) with 5% skim milk powder. Place the PVDF membrane in the primary antibody incubation solution and incubate overnight at 4°C in a shaking incubator.

[0124] Washing the membrane: After the primary antibody incubation is completed, remove the PVDF membrane and wash the membrane with 1× PBST on a shaker for 5 minutes each time for a total of 5 times;

[0125] Incubate with secondary antibody: Dilute Goat@Mouse IgG or Goat@Rabbit IgG corresponding to the primary antibody species in 5% skim milk powder at a dilution ratio of 1:2000. Place the rinsed PVDF membrane in the secondary antibody incubation solution and incubate on a shaking incubator at room temperature for 1 hour.

[0126] Wash the membrane: Take out the PVDF membrane incubated with secondary antibody and wash the membrane with 1×PBST on a shaker for 5 minutes each time for a total of 5 times;

[0127] Color development: Immerse the rinsed PVDF membrane in ECL luminescent color development solution, incubate at room temperature for 1 minute, and then use a Bio-Rad gel imager to develop, photograph, and analyze the expression level of PRRSV-N protein;

[0128] The results are as follows Figure 4 As shown, compared with the DMSO control group, the expression level of PRRSV-N protein was inhibited after MARC-145 cells were infected with PRRSV SD-YL1712 strain and treated with IDPA for 24h and 36h.

[0129] 5.TCID 50 Determination of PRRSV titer in supernatant samples

[0130] Plating: MARC-145 cells were plated at 1×10 4 Cells were seeded into 96-well cell plates at a density of 100 μL per well, and 100 μL of culture medium was added to each well. The cells were cultured in a 37°C incubator containing 5% CO2 for 12 h.

[0131] Dilute the virus: Use DMEM medium containing 3% serum to dilute the collected supernatant sample in a 10-fold gradient, from 10 -1 -10 -8 Three identical samples were diluted for each sample to perform biological replicates;

[0132] Inoculation: Discard the culture medium in the 96-well plate, add 100 μL of the serially diluted supernatant to each well of the cell plate, and continue culturing in a cell culture incubator at 37°C with 5% CO2.

[0133] IDPA treatment: Remove the 96-well plate, discard the virus solution, take 0.5 μL of IDPA stock solution and add it to 1 mL of DMEM medium containing 3% serum. Prepare the required IDPA working solution according to this concentration, mix well and add it to the 96-well plate. Incubate the plate in a cell culture incubator at 37°C with 5% CO2. For the control group, add DMSO.

[0134] Observe and record CPE: Observe continuously for 5 to 7 days and record the lesion condition in each well;

[0135] TCID 50 Calculation: Calculate the TCID of each test sample according to the Reed-Muench method 50 .

[0136] The results are as follows Figure 5 As shown, compared with the DMSO control group, after treatment with IDPA for 36 h in MARC-145 cells infected with PRRSV SD-YL1712 strain, the viral titer in the cell culture supernatant was reduced by about 3.94-log (P<0.05).

[0137] The above results show that the IDPA described in the present invention can significantly inhibit the infection and replication of the PRRSV SD-YL1712 strain and reduce the viral titer. It can be used to prepare drugs for preventing PRRSV infection or treating PRRS, providing a new treatment option for the prevention and treatment of PRRS and has broad application prospects.

[0138] In this specification, each embodiment is described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same and similar parts between the embodiments can be referred to each other. For the device disclosed in the embodiment, since it corresponds to the method disclosed in the embodiment, the description is relatively simple, and the relevant parts can be referred to the method part description. The above description of the disclosed embodiments enables professionals and technicians in this field to implement or use the present invention. Various modifications to these embodiments will be apparent to professionals and technicians in this field, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to these embodiments shown herein, but will comply with the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. An IDPA and its homologues, characterized in that: The structural formula of the IDPA and its homologues is shown in the following formula (I): Wherein, Ar is an aryl group, A is a straight chain or a 5- to 7-membered ring, R is hydrogen, an alkyl group or an aryl group, and R' is an alkyl group or an aryl group.

2. IDPA and its homologues according to claim 1, characterized in that The aryl group is selected from substituted or unsubstituted phenyl groups.

3. IDPA and its homologues according to claim 2, characterized in that The substituted phenyl groups include alkoxyphenyl groups, halogenated alkylphenyl groups, cyanophenyl groups, halogenated phenyl groups, and alkylphenyl groups.

4. IDPA and its homologues according to claim 3, characterized in that The A is a straight chain, and the R is a phenyl group.

5. IDPA and its homologues according to claim 3, characterized in that The A is a 5- to 7-membered carbon ring or a 5- to 7-membered oxygen heterocycle, and the R is hydrogen.

6. The use according to claim 3, characterized in that The IDPA and its homologues are selected from the compounds shown below:

7. Use of IDPA and its homologues according to any one of claims 1 to 6 in the preparation of antiviral drugs.

8. The use according to claim 7, characterized in that The virus is porcine reproductive and respiratory syndrome virus.

9. The use according to claim 7, characterized in that The IDPA and its homologues are added with pharmaceutically acceptable excipients to prepare any pharmaceutically acceptable dosage form.

10. The use according to claim 9, characterized in that The dosage forms include tablets, sprays, granules, capsules, oral solutions and injections.

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