Chiral 6-NH2 uracil compound as well as preparation method and application thereof

Abstract

The invention discloses a chiral 6-NH2 uracil compound as well as a preparation method and application thereof, and belongs to the technical field of chemical synthesis. The invention provides a method for constructing C-N axial chiral 6-NH2 uracil and a corresponding amide compound thereof through asymmetric organic catalysis, under the catalysis of chiral phosphoric acid, racemization 6-NH2 uracil and azlactone are used as reaction substrates, and 6-amide uracil with optical activity is obtained by utilizing a kinetic resolution strategy. And the 6-NH2 uracil compound with optical activity is recovered. According to the method, chiral uracil is constructed by using an asymmetric catalysis strategy for the first time, and the method has the characteristics of convenience in operation, cheap and easily available raw materials, and high yield and enantioselectivity. The C-N axial chiral uracil shows excellent anti-tumor cell activity, and especially has excellent biological activity on colon cancer, cervical cancer, multiple myeloma, prostate cancer cells and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of chemical synthesis, and in particular to a chiral 6-NH 2 Uracil compounds, preparation methods and applications thereof. Background Art

[0002] As a base unique to RNA, uracil is one of the basic components of vitamin B3, an important cofactor in RNA and terrestrial life metabolism. It replaces thymine in DNA during DNA transcription in living organisms and plays a very important role. Uracil can currently be used directly as a drug with diuretic, anti-inflammatory, and analgesic effects. Molecules containing uracil structures also have special drug activity and are widely used in the treatment of diseases. For example, 5-fluorouracil (Fluorouracil, 5-FU, also known as fluorine, fluorine, etc.), as an antimetabolite, is mainly used to treat tumors, such as digestive system cancer, breast cancer, ovarian cancer, cervical cancer, choriocarcinoma, malignant hydatidiform mole, bladder cancer, lung cancer, skin cancer, head and neck cancer, etc., and has been put into use. Uracil compounds containing N-aryl modifications have been studied in small quantities and have biological activity. For example, CX-659S is an anti-inflammatory drug with antioxidant activity, and its 6-aminouracil derivatives have significant effects on the treatment of atopic dermatitis (AD) and allergic contact dermatitis. GSK1120212 (DMSO solvate of JTP-74057) has been tested as a highly effective and selective inhibitor of mitogen-activated protein kinase / extracellular signal-regulated kinase (MEK). In addition, phosphodiesterase 4 (PDE 4) inhibitors with N-aryl uracil skeleton as the characteristic structure are under study and can be used to treat atopic dermatitis, psoriasis and chronic obstructive pulmonary disease. However, due to the limitations of the unique structure of uracil, the current synthesis methods for synthesizing CN-axial chiral N-aryl uracil are still limited to the resolution of chiral chromatographic columns and the introduction of chiral auxiliary groups or equivalent chiral bases. To date, the asymmetric catalytic synthesis of CN-axial chiral uracil has not been reported. Summary of the invention

[0003] In order to solve the above-mentioned deficiencies in the prior art, the present invention aims to provide a chiral 6-NH 2 Uracil compounds and preparation methods and applications thereof, the method uses an asymmetric organic catalytic strategy to obtain a series of 6-NH with CN axial chirality with high enantioselectivity 2 Uracil compounds and their corresponding amide derivatives. At the same time, this method has the characteristics of convenient operation, cheap and easy to obtain raw materials, mild reaction conditions, wide substrate universality and good product yield.

[0004] The technical solution of the present invention to solve the above technical problems is as follows: a chiral 6-NH 2Uracil compounds, the general structural formula is as follows:

[0005]

[0006] Among them, R 1 is a heterocyclic or substituted heterocyclic ring, an aryl or substituted aryl group; R 2 is a hydrogen atom, an aryl group or a substituted aryl group; R 3 is tert-butyl or halogen.

[0007] Furthermore, R 1 is phenyl, methoxy-substituted phenyl (ortho- or para-methoxy-substituted phenyl), halogen-substituted phenyl, trifluoromethyl-substituted phenyl (para-trifluoromethyl-substituted phenyl), thienyl, furyl, naphthyl; R 2 is a hydrogen atom, a phenyl group, a methyl group, a phenyl group substituted with a tert-butyl group, a phenyl group substituted with a methoxy group, or a phenyl group substituted with a halogen; R 3 is tert-butyl or halogen.

[0008] Furthermore, chiral 6-NH 2 The specific structural formula of uracil compounds is as follows:

[0009]

[0010] The chiral 6-NH 2 The preparation method of uracil compounds has the following reaction formula:

[0011]

[0012] Wherein, R is an aryl group or a substituted aryl group;

[0013] The specific preparation process is as follows:

[0014] The racemic uracil, azlactone, chiral phosphoric acid catalyst, additive and solvent are mixed and stirred at room temperature for 36-72 hours. After the reaction is completed, the chiral 6-NH 2 Uracil compounds.

[0015] Furthermore, racemic uracil, azlactone, chiral phosphoric acid catalyst, additive and solvent were mixed and stirred at room temperature for 36 hours.

[0016] Furthermore, the molar ratio of racemic uracil, azlactone and chiral phosphoric acid catalyst is 1:0.5-0.6:0.03-0.06.

[0017] Furthermore, the molar ratio of racemic uracil, azlactone and chiral phosphoric acid catalyst is 1:0.5-0.6:0.05.

[0018] Further, the structural formula of azlactone is as follows:

[0019]

[0020] Further, the structural formula of the chiral phosphoric acid catalyst is as follows:

[0021]

[0022] Furthermore, the solvent is dry dichloromethane, chloroform, carbon tetrachloride, 1,2-dichloroethane, ether, tetrahydrofuran, toluene or acetonitrile.

[0023] Furthermore, the additive is Molecular sieves, Molecular sieves, Molecular sieve or Na 2 SO 4 , the addition amount is 40-60 mg / mmol, preferably 50 mg / mmol.

[0024] The chiral 6-NH 2 Application of uracil compounds in the preparation of anti-tumor drugs.

[0025] Further, the tumor is renal cell adenocarcinoma, colon cancer, cervical cancer, multiple myeloma, prostate cancer and liver cancer.

[0026] The present invention has the following beneficial effects:

[0027] 1. The present invention provides an asymmetric organic catalytic construction of CN axial chiral 6-NH 2 The method of uracil and its corresponding amide compound is to use racemic 6-NH 2 Uracil and azlactone were used as reaction substrates, and the optically active 6-amidouracil was obtained by kinetic resolution strategy, and the optically active 6-NH 2 Uracil compound; This method is the first to use an asymmetric catalytic strategy to construct chiral uracil, and has the characteristics of easy operation, cheap and readily available raw materials, high yield and enantioselectivity.

[0028] 2. The synthesis method provided by the present invention shows excellent reactivity for various functional group-substituted uracil compounds, and has the characteristics of mild reaction conditions, wide substrate universality, good chemical selectivity and product yield.

[0029] 3. The CN-axis chiral uracil obtained by the present invention exhibits excellent anti-tumor cell activity, especially excellent biological activity against human renal cell adenocarcinoma, colon cancer, cervical cancer, multiple myeloma, prostate cancer and liver cancer cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 For chiral 6-NH 2 The results of the cytotoxicity experiment of uracil compound (S)-1j on human colon cancer cell HCT116.

[0031] Figure 2 For chiral 6-NH 2 The results of the cytotoxicity experiment of uracil compound (S)-1j on human cervical cancer cell HeLa.

[0032] Figure 3 For chiral 6-NH 2 The results of the cytotoxicity experiment of uracil compound (S)-1j on human multiple myeloma cells U266.

[0033] Figure 4 For chiral 6-NH 2 The results of the cytotoxicity experiment of uracil compound (S)-1j on human prostate cancer cell PC3.

[0034] Figure 5 6-NH 2 The results of cytotoxicity experiments of uracil compounds (S)-1j, (R)-1j and rac-1j on human renal cell adenocarcinoma cells ACHN.

[0035] Figure 6 6-NH 2 The results of cytotoxicity experiments of uracil compounds (S)-1j, (R)-1j and rac-1j on human prostate cancer cells LNCaP.

[0036] Figure 7 6-NH 2 The cytotoxicity experimental results of uracil compounds (S)-1j, (R)-1j and rac-1j on human liver cancer cell HepG2.

[0037] Figure 8 6-NH 2 The results of cytotoxicity experiments of uracil compounds (S)-1i, (R)-1i and rac-1i on human renal cell adenocarcinoma cells ACHN.

[0038] Fig. 9 6-NH 2 The results of cytotoxicity experiments of uracil compounds (S)-1i, (R)-1i and rac-1i on human colon cancer cells HCT116.

[0039] Fig.10 6-NH 2 The results of cytotoxicity experiments of uracil compounds (S)-1i, (R)-1i and rac-1i on human cervical cancer cells HeLa.

[0040] Fig.11 6-NH 2 The results of cytotoxicity experiments of uracil compounds (S)-1i, (R)-1i and rac-1i on human prostate cancer cells LNCaP.

[0041] Fig.12 6-NH 2 The results of cytotoxicity experiments of uracil compounds (S)-1i, (R)-1i and rac-1i on human multiple myeloma cells U266.

[0042] Fig.13 6-NH 2 The results of cytotoxicity experiments of uracil compounds (S)-1i, (R)-1i and rac-1i on human prostate cancer cells LNCaP.

[0043] Fig.14 6-NH 2 The results of cytotoxicity experiments of uracil compounds (S)-1i, (R)-1i and rac-1i on human liver cancer cells HepG2. DETAILED DESCRIPTION

[0044] The following examples are only used to explain the present invention and are not intended to limit the scope of the present invention. If no specific conditions are specified in the examples, the conditions are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0045] Embodiment 1:

[0046] A chiral 6-NH 2 The preparation method of uracil compounds comprises the following steps:

[0047] In a 5 mL reaction tube equipped with a stirrer, 0.1 mmol of racemic 6-NH 2 Uracil, 0.05mmol (0.5 equivalent) azlactone, 5mol% chiral phosphoric acid catalyst and 50mg activated Molecular sieves were added, 1 mL of dry dichloromethane was added, and stirred at room temperature for 36 h. After the reaction was completed, the optically active amide product was obtained by separation on a silica gel column, and the optically active 6-NH 2 Uracil.

[0048] The specific reaction process, products and corresponding yields are as follows:

[0049]

[0050]

[0051]

[0052] Note: Standard reaction conditions are: 0.1 mmol racemic uracil 1, 0.05 mmol (0.5 equivalent) of azlactone 2, chiral phosphoric acid catalyst (S)-A1 (5 mol%), Molecular sieve 50mg, ultra-dry dichloromethane solvent 1.0mL, react at room temperature for 36h. The yield is the isolated yield, and the corresponding selectivity er (enantiomeric ratio) value is measured by HPLC. The selectivity factor s is calculated as follows: s = ln [(1-C)(1-ee sub )] / ln[(1-C)(1+ee sub )],C=ee sub / (ee pro +ee sub );where ee sub To recover the enantiomeric excess of raw material (R)-1, ee pro The enantiomeric excess of product (S)-3.

[0053] The chiral phosphoric acid catalyst (S)-A1 used has the following structural formula:

[0054]

[0055] The above experimental results show that uracil modified with different substituents, whether electron-rich groups, electron-poor groups or neutral groups, is compatible and can obtain the target acylated product 3 with excellent yield and enantioselectivity, and recover the optically active 6-NH 2 Uracil compounds. Various functional groups or substituents are compatible in this reaction system, including halogen, alkyl, methoxy, heteroatom-containing rings, condensed rings, etc.

[0056] The product prepared by the present invention was subjected to nuclear magnetic resonance and mass spectrometry characterization analysis, and the nuclear magnetic resonance and mass spectrometry characterization data results were consistent with the obtained product. The specific characterization data are as follows:

[0057] (R)-6-amino-3-benzyl-1-(2-(tert-butyl)phenyl)pyrimidine-2,4(1H,3H)-dione(1a)

[0058]

[0059] Yield: 17.12 mg, yield: 49%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2;

[0060] 1H NMR(400MHz,Chloroform-d)δ7.61(dd,J=8.2,1.5Hz,1H),7.45-7.40(m,1H),7.39-7.35(m,2H),7.30(td,J=7.6,1.5Hz,1H),7.1 9(t,J=7.2Hz,2H),7.15-7.10(m,1H),7.00(dd,J=7.8,1.5Hz,1H),5.11(s,1H),5.05(d,J=20.0Hz,2H),4.72(s,2H),1.21(s,9H); 13 C NMR(101MHz,Chloroform-d)δ163.3,154.1,151.8,148.6,137.5,131.2,130.8, 130.42,130.38,128.7,128.1,128.0,127.2,44.0,36.2,31.4; HRMS(ESI):calcd forC 21 H 24 N 3 O 2 [M+H] + :350.1863,found:350.1861.

[0061] (R)-6-amino-3-benzyl-1-(3-(tert-butyl)-[1,1'-biphenyl]-4-yl)pyrimidine-2,4(1H,3H)-dione(1b)

[0062]

[0063] Yield: 18.30 mg, yield: 42%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2;

[0064] 1 H NMR(400MHz,Chloroform-d)δ7.73(d,J=2.1Hz,1H),7.64-7.59(m,2H),7.50(dd,J=8.1,2.1Hz,1H),7.40(dd,J=8.4,6.9Hz,2H),7.34-7. 28(m,1H),7.18(d,J=3.8Hz,4H),7.14-7.11(m,1H),7.08(d,J=8.1Hz,1H),6.25(s,2H),4.83(d,J=14.8Hz,2H),4.79(s,1H),1.15(s,9H); 13C NMR (151 MHz, DMSO-d 6 )δ161.6,155.0,151.7,148.1,141.3,139.7,138.1,132.2,130.9,129.1,128.14, 128.08,127.8,127.6,127.0,126.9,125.9,74.7,42.9,35.9,31.1; HRMS(ESI):m / z calcd for C 27 H 28 N 3 O 2 [M+H] + :426.2176,found:426.2171.

[0065] (R)-6-amino-3-benzyl-1-(3-(tert-butyl)-4'-methyl-[1,1'-biphenyl]-4-yl)pyrimidine-2,4(1H,3H)-dione(1c)

[0066]

[0067] Yield: 18.30 mg, yield: 42%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2;

[0068] 1 H NMR(400MHz,Chloroform-d)δ7.78(d,J=2.1Hz,1H),7.51-7.42(m,5H),7.30-7.17(m,5H),7.08( d,J=8.1Hz,1H),5.15(d,J=2.4Hz,1H),5.14-5.01(m,2H),4.52(s,2H),2.41(s,3H),1.26(s,9H); 13 C NMR(101MHz,Chloroform-d)δ153.7,151.8,148.8,143.4,138.0,137.6,137.3,131.5,129.7, 129.2,129.0,128.2,127.3,127.2,126.6,100.0,77.7,44.2,36.4,31.5,21.2; HRMS(ESI):m / z calcd for C 28 H 30 N 3 O 2 [M+H] +:440.2333found:440.2333.

[0069] (R)-6-amino-3-benzyl-1-(3,3'-di-tert-butyl-[1,1'-biphenyl]-4-yl)pyrimidine-2,4(1H,3H)-dione(1d)

[0070]

[0071] Yield: 22.64 mg, yield: 47%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2;

[0072] 1 H NMR(400MHz,Chloroform-d)δ7.78(d,J=2.1Hz,1H),7.55(d,J=1.8Hz,1H),7.50(dd,J=8.1,2.1Hz,1H),7.45-7.34(m,5H),7.23(dd,J= 13.7,6.9Hz,2H),7.17(d,J=7.1Hz,1H),7.09(d,J=8.0Hz,1H),5.17(s,1H),5.13-5.00(m,2H),4.75(s,2H),1.38(s,9H),1.27(s,9H); 13 C NMR(101MHz,Chloroform-d)δ163.4,154.0,151.9,151.8,148.7,144.1,140.1,137.6,131.5,129.7, 129.6,128.9,128.7,128.1,127.2,127.0,125.0,124.5,44.1,36.4,34.8,31.5,31.4.HRMS(ESI):m / z calcd for C 31 H 36 N 3 O 2 [M+H] + :482.2802 found:482.2801.

[0073] (R)-6-amino-3-benzyl-1-(3-(tert-butyl)-4'-methoxy-[1,1'-biphenyl]-4-yl)pyrimidine-2,4(1H,3H)-dione(1e)

[0074]

[0075] Yield: 22.32 mg, yield: 49%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2;

[0076] 1 H NMR(400MHz,Chloroform-d)δ7.74(d,J=2.1Hz,1H),7.52-7.47(m,2H),7.44(dd,J=8.1,2.1Hz,1H),7.39(d,J=7.2Hz,2H),7.18(t,J=7.3Hz,2H ),7.14-7.07(m,1H),7.03(d,J=8.1Hz,1H),7.01-6.91(m,2H),5.15(s, 1H),5.14-4.96(m,2H),4.85(d,J=8.0Hz,2H),3.84(s,3H),1.25(s,9H); 13 CNMR(101MHz,Chloroform-d)δ163.3,159.6,154.2,151.9,148.6,142.8,137.6,132.6,131.5,129 .2,128.8,128.7,128.3,128.1,127.1,126.3,114.4,77.2,55.4,44.1,36.3,31.5; HRMS(ESI):m / z calcd for C 28 H 30 N 3 O 3 [M+H] + :456.2282 found:456.2280.

[0077] (R)-6-amino-3-benzyl-1-(3-(tert-butyl)-4'-fluoro-[1,1'-biphenyl]-4-yl)pyrimidine-2,4(1H,3H)-dione(1f)

[0078]

[0079] Yield: 21.73 mg, yield: 49%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2;

[0080] 1H NMR(400MHz,Chloroform-d)δ7.74(d,J=2.1Hz,1H),7.55-7.48(m,2H),7.45(dd,J=8.1,2.1Hz,1H),7.43-7.37(m,2H),7.23-7. 18(m,2H),7.17-7.14(m,2H),7.12(s,1H),7.08(d,J=8.1Hz,1H),5.17(s,1H),5.07(d,J=17.9Hz,2H),4.80(s,2H),1.26(s,9H); 13 C NMR(101MHz,Chloroform-d)δ164.1,163.3,161.6,152.9(d,J=219.2Hz),149.0,142.4,137.5,136.3(d,J=3 .4Hz),131.7,129.9,129.2,128.9(d,J=2.5Hz),128.1,127.2,126.7,115.9(d,J=21.5Hz),44.1,36.4,31.5; 19 F NMR(376MHz,Chloroform-d)δ-114.37; HRMS(ESI):m / z calcd for C 27 H 27 FN 3 O 2 [M+H] + :444.2082found:444.2080.

[0081] (R)-6-amino-1-(2-(tert-butyl)phenyl)-3-(4-methoxybenzyl)pyrimidine-2,4(1H,3H)-dione(1g)

[0082]

[0083] Yield: 18.59 mg, yield: 49%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2; 1H NMR(400MHz,Chloroform-d)δ7.62(dd,J=8.2,1.4Hz,1H),7.48-7.36(m,3H),7.31(d,J=1.4Hz,1H),7.03(dd,J= 7.7,1.5Hz,1H),6.86-6.66(m,2H),5.10(s,1H),5.01(d,J=11.7Hz,2H),4.43(s,2H),3.76(s,3H),1.22(s,9H); 13 C NMR(101MHz,Chloroform-d)δ163.2,158.8,153.5,151.8,148.7,131.2,130.8,130 .6,130.5,130.4,129.8,128.0,113.5,77.7,55.2,43.5,36.2,31.4; HRMS(ESI):m / z calcd for C 22 H 26 N 3 O 3 [M+H] + :380.1969found:380.1971.

[0084] (R)-6-amino-1-(2-(tert-butyl)phenyl)-3-(2-methoxybenzyl)pyrimidine-2,4(1H,3H)-dione(1h)

[0085]

[0086] Yield: 18.59 mg, yield: 49%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2; 1 H NMR(400MHz,Chloroform-d)δ7.59(d,J=8.0Hz,1H),7.41(t,J=7.6Hz,1H),7.34-7.22(m,1H),7.07(t,J=7.8Hz,1H ),6.99(t,J=7.6Hz,2H),6.81-6.67(m,2H),5.15(s,1H),5.14-5.00(m,2H),4.72(s,2H),3.76(s,3H),1.24(s,9H); 13C NMR(101MHz,Chloroform-d)δ156.9,154.3,151.7,148.6,131.3,130.9,130.34,130.27 ,127.9,127.7,126.7,125.5,120.1,110.1,77.1,55.4,39.4,36.2,31.5; HRMS(ESI):m / z calcd for C 22 H 26 N 3 O 3 [M+H] + :380.1969found:380.1962.

[0087] (R)-6-amino-1-(2-(tert-butyl)phenyl)-3-(3,4-dichlorobenzyl)pyrimidine-2,4(1H,3H)-dione(1i)

[0088]

[0089] Yield: 20.08 mg, yield: 48%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2; 20.08mg, 48% yield; 1 H NMR(400MHz,Chloroform-d)δ7.67-7.62(m,1H),7.56(d,J=1.4Hz,1H),7.46(td,J=8.2,7.7,1.5Hz,1H),7. 37-7.30(m,3H),7.06(dd,J=7.8,1.5Hz,1H),5.11(s,1H),5.02(d,J=15.3Hz,2H),4.43(s,2H),1.24(s,9H); 13 C NMR(101MHz,Chloroform-d)δ162.8,153.6,151.7,148.7,137.7,132.2,131.4,131.2,13 1.1,130.7,130.60,130.55,130.2,128.7,128.1,77.6,43.0,36.3,31.4; HRMS(ESI):m / z calcd for C 21 H 22 Cl 2 N 3 O 2 [M+H] +:418.1084 found:418.1082.

[0090] (R)-6-amino-1-(2-(tert-butyl)phenyl)-3-(3,5-difluorobenzyl)pyrimidine-2,4(1H,3H)-dione(1j)

[0091]

[0092] Yield: 18.88 mg, yield: 49%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2; 1 H NMR(400MHz,Chloroform-d)δ7.64(dd,J=8.2,1.4Hz,1H),7.48-7.41(m,1H),7.33(td,J=7.5,1.5Hz,1H),7.05(dd,J=7.8 ,1.5Hz,1H),6.93(h,J=4.3Hz,2H),6.64(tt,J=9.0,2.4Hz,1H),5.12(s,1H),5.10-4.92(m,2H),4.68(s,2H),1.24(s,9H); 13 C NMR(101MHz,Chloroform-d)δ162.9,162.8(d,J=248.1Hz),154.0,151.7,148.6,141.2(d,J=9.2Hz),1 31.1,130.62,130.56,130.5,128.1,111.6(d,J=25.4Hz),102.8(d,J=25.3Hz),77.2,43.4,36.2,31.4; 19 F NMR(376MHz,Chloroform-d)δ-62.55; HRMS(ESI):m / z calcd forC 21 H 22 F 2 N 3 O 2 [M+H] + :386.1675found:386.1673.

[0093] (R)-6-amino-1-(2-(tert-butyl)phenyl)-3-(4-(trifluoromethyl)benzyl)pyrimidine-2,4(1H,3H)-dione(1k)

[0094]

[0095] Yield: 16.70 mg, yield: 40%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2; 1 H NMR(400MHz,Chloroform-d)δ7.61(dd,J=8.2,1.5Hz,1H),7.50(t,J=5.6Hz,4H),7.44-7.38(m,1H),7.30(td,J=7.5 ,1.5Hz,1H),7.00(dd,J=7.8,1.6Hz,1H),5.15(d,J=14.0Hz,2H),5.05(d,J=14.1Hz,1H),4.86(s,2H),1.19(s,9H); 13 C NMR(101MHz,Chloroform-d)δ163.0,154.2,151.7,148.5,141.4,131.1,130.5,129.4(d,J=32 .2Hz), 128.5 (d, J = 98.3Hz), 125.1 (d, J = 3.7Hz), 124.1 (q, J = 272.0Hz), 77.0, 43.6, 36.1, 31.3; 19 FNMR(376MHz,Chloroform-d)δ-62.48; HRMS(ESI):m / z calcd for C 22 H 23 F 3 N 3 O 2 [M+H] + :418.1737 found:418.1736.

[0096] (R)-6-amino-3-(4-bromobenzyl)-1-(2-(tert-butyl)phenyl)pyrimidine-2,4(1H,3H)-dione(1l)

[0097]

[0098] Yield: 20.99 mg, yield: 49%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2; 1H NMR(400MHz,Chloroform-d)δ7.62(d,J=1.5Hz,1H),7.48-7.38(m,1H),7.38-7.26(m,4H),7.14(s ,1H),7.02(dd,J=7.8,1.5Hz,1H),5.06(s,1H),5.00(d,J=17.5Hz,2H),4.59(s,2H),1.22(s,9H); 13 C NMR(101MHz,Chloroform-d)δ163.1,153.8,151.8,148.6,136.4,131.9,131.3,1 31.1,130.8,130.6,130.5,128.1,121.3,100.0,43.4,36.2,31.4; HRMS(ESI):m / z calcd for C 21 H 23 Bn 3 O 2 [M+H] + :428.0968found:428.0961.

[0099] (R)-6-amino-1-(2-(tert-butyl)phenyl)-3-(thiophen-2-ylmethyl)pyrimidine-2,4(1H,3H)-dione(1m)

[0100]

[0101] Yield: 16.71 mg, yield: 47%, properties: yellow solid; R f (PE / EA=1:1)=0.1-0.2; 1 H NMR(400MHz,Chloroform-d)δ7.63(dd,J=8.2,1.5Hz,1H),7.43(td,J=7.7,1.5Hz,1H),7.32(dd,J=7.6,1.5Hz,1H),7.14-7.07 (m,2H),7.03(dd,J=7.8,1.5Hz,1H),6.83(dd,J=5.1,3.5Hz,1H),5.21(d,J=3.4Hz,2H),5.10(s,1H),4.74(s,2H),1.24(s,9H); 13C NMR (101MHz, Chloroform-d) δ162.7,154.0,151.5,148.7,139.0,131.2,130.7,130.5,130.4,128.0,126.2,125.4,77.3,38.5,36.2,31.5; HRMS (ESI): m / z calcd for C 19 H 22 N 3 O 2 S[M+H] + :356.1427 found:356.1421.

[0102] (R)-6-amino-1-(2-(tert-butyl)phenyl)-3-(furan-2-ylmethyl)pyrimidine-2,4(1H,3H)-dione(1n)

[0103]

[0104] Yield: 15.95 mg, yield: 47%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2; 1 H NMR(400MHz,Chloroform-d)δ7.64(dd,J=8.2,1.5Hz,1H),7.44(ddd,J=8.3,7.3,1.6Hz,1H),7.39-7.30(m,1H),7.24(dd,J=1.9,0.9Hz,1H),7.0 6(dd,J=7.7,1.6Hz,1H),6.28(d,J=3.2Hz,1H),6.23(dd,J=3.2,1.9Hz,1 H),5.18-5.09(m,2H),5.04(d,J=14.8Hz,1H),4.62(s,2H),1.25(s,9H); 13 C NMR(101MHz,Chloroform-d)δ162.8,153.9,151.5,150.6,148.7,141.7,131.2 ,130.7,130.49,130.47,128.0,110.2,108.8,37.0,36.3,31.4; HRMS(ESI):m / z calcd for C 19 H 22 N 3 O 3 [M+H] + :340.1656found:340.1656.

[0105] (R)-6-amino-1-(2-(tert-butyl)phenyl)-3-(naphthalen-2-ylmethyl)pyrimidine-2,4(1H,3H)-dione(1o)

[0106]

[0107] Yield: 18.78 mg, yield: 47%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2; 1 H NMR(400MHz,Chloroform-d)δ7.87(d,J=1.6Hz,1H),7.71(s,3H),7.63-7.52(m,2H),7.45-7.35(m,3H),7. 30-7.20(m,1H),6.96(dd,J=7.8,1.5Hz,1H),5.23(d,J=4.9Hz,2H),5.15(s,1H),4.55(s,2H),1.18(s,9H); 13 CNMR(101MHz,Chloroform-d)δ163.2,153.8,151.8,148.6,135.0,133.3,132.7,131.2,130.8,130. 4,128.0,127.9,127.79,127.76,127.5,127.0,125.8,125.6,77.6,44.2,36.2,31.4; HRMS(ESI):m / z calcd for C 25 H 26 N 3 O 2 [M+H] + :400.2020found:400.2018.

[0108] (R)-6-amino-3-benzyl-1-(2-iodophenyl)pyrimidine-2,4(1H,3H)-dione(1p)

[0109]

[0110] Yield: 20.54 mg, yield: 49%, properties: white solid; R f (PE / EA=1:1)=0.1-0.2; 11H NMR (400 MHz, Chloroform-d) δ 7.98 (d, J = 8.0 Hz, 1H), 7.51 (t, J = 7.7 Hz, 1H), 7.39 (d, J = 7.5 Hz, 2H), 7.33 (d, J = 7.8 Hz, 1H), 7.21 (t, J = 7.8 Hz, 3H), 7.14 (d, J = 7.3 Hz, 1H), 5.11 (s, 1H), 5.07 (d, J = 11.5 Hz, 2H), 4.59 (s, 2H); 13 13C NMR (101 MHz, Chloroform-d) δ 163.2, 152.6, 150.6, 140.6, 137.5, 136.7, 131.7, 130.4, 130.2, 128.4, 128.2, 127.2, 99.9, 77.5, 44.2; HRMS (ESI): m / z calcd for C 17 H 15 IN 3 O 2 [M + H] + : 420.0204 found: 420.0201.

[0111] (S)-N-(2-((1-benzyl-3-(2-(tert-butyl)phenyl)-2,6-dioxo-1,2,3,6-tetrahydropy-rimidin-4-yl)amino)-2-oxoethyl)benzamide (3a)

[0112]

[0113] Yield: 25.02 mg, Yield rate: 49%, Appearance: white solid; R f (PE / EA = 1:1) = 0.4 - 0.5; 1 1H NMR (400 MHz, Chloroform-d) δ 11.01 (s, 1H), 7.89 - 7.79 (m, 2H), 7.66 (d, J = 1.5 Hz, 1H), 7.49 (td, J = 7.7, 1.6 Hz, 4H), 7.46 - 7.33 (m, 3H), 7.33 - 7.21 (m, 4H), 7.05 (dd, J = 7.8, 1.5 Hz, 1H), 5.21 (s, 1H), 5.18 - 5.06 (m, 2H), 4.99 (dd, J = 12.4, 5.0 Hz, 2H), 1.20 (s, 9H); 13C NMR(101MHz,Chloroform-d)δ194.6,167.1,161.1,158.6,149.9,148.5,136.9,131.3,131.1,131.0, 130.9,129.3,129.2,128.6,128.5,128.3,127.6,127.1,90.2,50.5,44.3,36.2,31.4; HRMS(ESI):m / z calcd for C 30 H 31 N 4 O 4 [M+H] + :511.2340 found:511.2339.

[0114] (S)-N-(2-((1-benzyl-3-(3-(tert-butyl)-[1,1'-biphenyl]-4-yl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3b)

[0115]

[0116] Yield: 28.75 mg, yield: 49%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR(400MHz,Chloroform-d)δ11.06(s,1H),7.84(dq,J=4.8,2.3Hz,3H),7.55(dt,J=7.2,2.2Hz,3H),7.53-7.44(m,6H),7.41(dd,J=6.7,2.3Hz,2H),7.2 8(dtd,J=13.8,7.4,6.9,3.5Hz,4H),7.12(dd,J=8.1,2.5Hz,1H),5.25-5.22 (m,1H),5.22-5.09(m,2H),5.02(ddd,J=12.0,5.1,2.5Hz,2H),1.26(s,9H); 13C NMR(101MHz,Chloroform-d)δ194.7,167.2,161.1,158.7,150.0,148.8,144.1,139.9,136.9,134.7,131.44,131.37,12 9.9,129.4,129.0,128.5,128.4,128.3,128.2,127.7,127.4,127.3,127.1,90.2,50.6,44.4,36.4,31.5; HRMS(ESI):m / z calcd for C 36 H 35 N 4 O 4 [M+H] + :587.2653found:587.2653.

[0117] (S)-N-(2-((1-benzyl-3-(3-(tert-butyl)-4'-methyl-[1,1'-biphenyl]-4-yl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3c)

[0118]

[0119] Yield: 24.03 mg, yield: 40%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR(400MHz,Chloroform-d)δ10.96(d,J=4.1Hz,1H),7.84-7.66(m,3H),7.49-7.28(m,7H),7.20(dd,J=7.9,2. 3Hz,7H),7.03(s,1H),5.15(s,1H),5.13-5.02(m,2H),4.92(dd,J=10.8,5.0Hz,2H),2.33(s,3H),1.17(s,9H); 13C NMR(101MHz,Chloroform-d)δ194.5,167.2,161.1,158.7,150.0,148.7,144.0,138.2,137.0,136.9,134.6,131.4,1 29.8,129.6,129.4,128.5,128.4,127.9,127.7,127.13,127.08,90.2,50.5,44.4,36.4,31.5,21.2; HRMS(ESI):m / z calcd for C 37 H 37 N 4 O 4 [M+H] + :601.2810 found:601.2814.

[0120] (S)-N-(2-((1-benzyl-3-(3,3'-di-tert-butyl-[1,1'-biphenyl]-4-yl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3d)

[0121]

[0122] Yield: 30.21 mg, yield: 47%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR(400MHz,Chloroform-d)δ11.23-10.80(m,1H),7.91-7.85(m,2H),7.83(d,J=2.0Hz,1H),7.59-7.50(m,4H),7.49-7.39(m,5H),7.37(s,1H), 7.33-7.24(m,4H),7.12(d,J=8.1Hz,1H),5.22(d,J=13.8Hz,1H),5.14(d ,J=13.8Hz,2H),5.03(dd,J=12.2,5.0Hz,2H),1.38(s,9H),1.26(s,9H); 13C NMR(101MHz,Chloroform-d)δ194.7,167.2,161.1,158.7,152.0,150.0,148.7,144.9,139.8,136.9,134.7,131.4,130.1,129.5 ,128.8,128.5,128.4,128.1,127.7,127.6,127.1,125.3,124.6,100.0,90.3,50.6,44.4,36.4,34.9,31.5,31.4; HRMS(ESI):m / z calcd for C 40 H 43 N 4 O 4 [M+H] + :643.3279found:643.3279.

[0123] (S)-N-(2-((1-benzyl-3-(3-(tert-butyl)-4'-methoxy-[1,1'-biphenyl]-4-yl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3e)

[0124]

[0125] Yield: 29.60 mg, yield: 48%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR(400MHz,Chloroform-d)δ11.04(d,J=4.1Hz,1H),7.91-7.82(m,2H),7.80(d,J=2.1Hz,1H),7.58-7.46(m,6H),7.42(dd,J=8.3,6.5Hz,2H),7.36 -7.22(m,4H),7.09(d,J=8.1Hz,1H),6.99(d,J=8.3Hz,2H),5.23(s,1H),5 .21-5.08(m,2H),5.01(dd,J=11.2,5.0Hz,2H),3.86(s,3H),1.25(s,9H); 13C NMR(101MHz,Chloroform-d)δ194.6,167.1,161.1,159.8,158.7,150.0,148.6,143.6,136.9,134.6,132.3,131.4,129. 4,129.3,128.5,128.4,128.3,127.67,127.63,127.1,126.9,114.4,90.2,55.4,50.5,44.4,36.4,31.5; HRMS(ESI):m / z calcd for C 37 H 37 N 4 O 5 [M+H] + :617.2759found:617.2763.

[0126] (S)-N-(2-((1-benzyl-3-(3-(tert-butyl)-4'-fluoro-[1,1'-biphenyl]-4-yl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3f)

[0127]

[0128] Yield: 25.40 mg, yield: 42%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR(400MHz,Chloroform-d)δ10.99(d,J=4.1Hz,1H),7.82-7.74(m,2H),7.72(d,J=2.0Hz,1H),7.47-7.40(m,6H),7.36(d,J=7.5Hz, 2H),7.25-7.17(m,4H),7.13-7.02(m,3H),5.14(d,J=14.0Hz,1H),5.05(d,J=13.8Hz,2H),4.94(dd,J=10.9,5.0Hz,2H),1.18(s,9H); 13CNMR(101MHz,Chloroform-d)δ194.6,167.2,161.7,159.8(d,J=242.0Hz),150.0,149.0,143.1,136.8,136.0(d,J=3.2Hz),134.6,131.6 ,131.4,129.7,129.4,129.0(d,J=8.1Hz),128.5,128.4,128.3,127.7,127.2,127.1,116.0(d,J=21.4Hz),90.2,50.5,44.4,36.4,31.5; 19 F NMR(376MHz,Chloroform-d)δ-113.98; HRMS(ESI):m / z calcd forC 36 H 34 FN 4 O 4 [M+H] + :605.2557found:605.2562.

[0129] (S)-N-(2-((3-(2-(tert-butyl)phenyl)-1-(4-methoxybenzyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3g)

[0130]

[0131] Yield: 21.62 mg, yield: 40%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR (400MHz, Chloroform-d) δ10.94(s,1H),7.78(dt,J=8.5,1.7Hz,2H),7.61(dt,J=8.2,1.7Hz,1H),7.48-7.32(m,6H),7.29-7.14(m,2H),6.98(dt,J= 7.8,1.7Hz,1H),6.80-6.65(m,2H),5.03(d,J=1.6Hz,1H),5.03-4.95(m,2H ), 4.93 (dd, J = 5.0, 1.7Hz, 2H), 3.71 (d, J = 1.7Hz, 3H), 1.14 (d, J = 1.7Hz, 9H); 13C NMR(101MHz,Chloroform-d)δ194.7,167.2,161.1,159.2,158.6,149.9,148.6,131.4,131.12,131.09,131 .01,130.95,129.3,129.2,128.6,128.5,127.1,113.7,90.3,55.3,50.5,43.8,36.3,31.5; HRMS(ESI):m / z calcd forC 31 H 33 N 4 O 5 [M+H] + :541.2446found:541.2449.

[0132] (S)-N-(2-((3-(2-(tert-butyl)phenyl)-1-(2-methoxybenzyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3h)

[0133]

[0134] Yield: 21.62 mg, yield: 40%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 21.62mg, 40% yield; 1 H NMR(400MHz,Chloroform-d)δ11.08-10.89(m,1H),7.80-7.72(m,2H),7.59(dd,J =8.2,1.5Hz,1H),7.44-7.38(m,2H),7.34(dd,J=14.7,7.8Hz,2H),7.32-7.25(m,1 H),7.18-7.10(m,2H),7.05-6.97(m,2H),6.78(t,J=7.8Hz,2H),5.22(d,J=15.3H z,1H),5.13-4.96(m,2H),4.91(dd,J=10.3,5.1Hz,2H),3.76(s,3H),1.17(s,9H); 13C NMR(101MHz,Chloroform-d)δ194.8,167.1,161.2,158.8,157.2,149.7,148.6,134.7,131.3,131.1,131.0,130.9, 129.3,128.6,128.5,128.2,127.6,127.1,124.7,120.3,110.4,90.2,55.5,50.5,39.9,36.3,31.5; HRMS(ESI):m / z calcd for C 31 H 33 N 4 O 5 [M+H]+:541.2446found:541.2448.

[0135] (S)-N-(2-((3-(2-(tert-butyl)phenyl)-1-(3,4-dichlorobenzyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3i)

[0136]

[0137] Yield: 28.39 mg, yield: 49%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5;, 28.39mg, 49% yield; 1 H NMR(400MHz,Chloroform-d)δ10.99(d,J=3.9Hz,1H),7.82-7.74(m,2H),7.63 (dd,J=8.3,1.5Hz,1H),7.53(d,J=1.2Hz,1H),7.48-7.41(m,2H),7.39-7.32(m ,3H),7.29(d,J=1.2Hz,2H),7.19-7.12(m,1H),6.99(dd,J=7.8,1.5Hz,1H),5 .05(d,J=13.9Hz,1H),5.00(s,2H),4.92(dd,J=10.6,5.0Hz,2H),1.15(s,9H); 13C NMR(101MHz,Chloroform-d)δ194.6,167.2,160.8,158.7,149.9,148.6,137.0,134.6,132.4,131.9,131.44,1 31.37,131.3,131.03,130.99,130.3,129.0,128.7,128.5,127.1,90.1,50.5,43.3,36.3,31.4; HRMS(ESI):m / z calcd for C 30 H 29 Cl 2 N 4 O 4 [M+H] + :579.1561found:579.1565.

[0138] (S)-N-(2-((3-(2-(tert-butyl)phenyl)-1-(3,5-difluorobenzyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3j)

[0139]

[0140] Yield: 26.78 mg, yield: 49%, properties: yellow solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR(400MHz,Chloroform-d)δ11.19-10.64(m,1H),7.79-7.72(m,2H),7.62(dd,J=8 .3,1.5Hz,1H),7.46-7.43(m,1H),7.42-7.39(m,1H),7.39-7.29(m,3H),7.15(t,J=4 .6Hz,1H),6.99(dd,J=7.8,1.5Hz,1H),6.97-6.91(m,2H),6.67-6.59(m,1H),5.08( d,J=13.9Hz,1H),4.98(d,J=13.9Hz,2H),4.91(dd,J=10.7,5.0Hz,2H),1.15(s,9H); 13C NMR(101MHz,Chloroform-d)δ194.5,167.2,162.8(d,J=248.5Hz),160.8,158.7,149.9,148.5,140.5(d,J=9.2Hz),134.6, 131.4,131.3,131.0,129.0,128.7,128.5,127.1,112.1(d,J=25.4Hz),103.2(d,J=50.5Hz),90.0,50.5,43.6,36.3,31.4; 19 F NMR(376MHz,Chloroform-d)δ-109.79; HRMS(ESI):m / z calcd for C 30 H 29 F 2 N 4 O 4 [M+H] + :547.2152 found:547.2157.

[0141] (S)-N-(2-((3-(2-(tert-butyl)phenyl)-2,6-dioxo-1-(4-(trifluoromethyl)benzyl)-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3k)

[0142]

[0143] Yield: 27.19 mg, yield: 47%, properties: yellow solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR(400MHz,Chloroform-d)δ11.06(s,1H),7.89-7.82(m,2H),7.70(dd,J=8.1,1.5Hz,1H),7.62(d,J=8.1Hz,2H),7.58-7.48(m,4H),7.47- 7.36(m,3H),7.23(d,J=4.3Hz,1H),7.06(dd,J=7.9,1.5Hz,1H),5.29-5.12(m,2H),5.07(s,1H),5.01(dd,J=11.7,5.0Hz,2H),1.21(s,9H); 13C NMR (101MHz, Chloroform-d) δ194.4,167.2,161.0,158.7,149.9,148.4,140.7,134.5,131.4,131.2,130.9(d,J=2.4Hz),129.8( d,J=32.5Hz),129.6,129.0,128.6,128.5,128.0,127.0,125.3(d,J=3.7Hz),124.1(q,J=272.0Hz),90.0,50.5,43.8,36.2,31.4; 19 F NMR(376MHz,Chloroform-d)δ-62.55; HRMS(ESI):m / z calcd forC 31 H 30 F 3 N 4 O 4 [M+H] + :579.2214found:579.2219.

[0144] (S)-N-(2-((1-(4-bromobenzyl)-3-(2-(tert-butyl)phenyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3l)

[0145]

[0146] Yield: 28.88 mg, yield: 49%, properties: yellow solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR(400MHz,Chloroform-d)δ11.04(d,J=4.0Hz,1H),7.89-7.82(m,2H),7.69(dd,J=8.3,1.5Hz,1H),7.54-7.47(m,2H),7.46-7.37(m,7H) ,7.28-7.22(m,1H),7.05(dd,J=7.8,1.5Hz,1H),5.14(d,J=13.8Hz,1H),5.07(d,J=2.1Hz,2H),5.00(dd,J=11.3,5.0Hz,2H),1.21(s,9H); 13C NMR(101MHz,Chloroform-d)δ194.6,167.2,161.0,158.7,149.9,148.6,135.8,134.6,131.5,131.4,131. 3,131.2,131.02,130.97,129.1,128.6,128.5,127.1,121.8,90.1,50.5,43.7,36.3,31.4; HRMS(ESI):m / z calcd for C 30 H 30 Bn 4 O 4 [M+H] + :589.1445found:589.1448.

[0147] (S)-N-(2-((3-(2-(tert-butyl)phenyl)-2,6-dioxo-1-(thiophen-2-ylmethyl)-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3m)

[0148]

[0149] Yield: 25.31 mg, yield: 49%, properties: yellow solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR(400MHz,Chloroform-d)δ11.02(d,J=4.0Hz,1H),7.87-7.81(m,2H),7.69(dd,J=8.2,1.5Hz,1H),7.54-7.46(m,2H),7.45-7.35(m,3H),7.26-7.1 5(m,3H),7.06(dd,J=7.9,1.5Hz,1H),6.92(dd,J=4.8,3.7Hz,1H),5.36-5. 26(m,2H),5.10(d,J=3.8Hz,1H),5.00(dd,J=8.4,5.0Hz,2H),1.24(s,9H); 13C NMR(101MHz,Chloroform-d)δ194.6,167.2,160.6,158.7,149.6,148.6,138.1,134.7,131.4,131.2,131. 05,130.98,129.1,128.7,128.6,128.5,127.1,126.4,125.9,90.2,50.5,38.8,36.3,31.5; HRMS(ESI):m / z calcdfor C 28 H 29 N 4 O 4 S[M+H] + :517.1904 found:517.1910.

[0150] (S)-N-(2-((3-(2-(tert-butyl)phenyl)-1-(furan-2-ylmethyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3n)

[0151]

[0152] Yield: 23.02 mg, yield: 46%, properties: yellow solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR(400MHz,Chloroform-d)δ11.05(s,1H),7.90-7.79(m,2H),7.70(dd,J=8.2,1.5 Hz,1H),7.54-7.47(m,2H),7.45-7.36(m,3H),7.32(dd,J=1.9,0.9Hz,1H),7.26-7.1 7(m,1H),7.07(dd,J=7.8,1.5Hz,1H),6.41-6.35(m,1H),6.30(dd,J=3.2,1.8Hz,1H) ,5.28-5.12(m,2H),5.06(d,J=4.3Hz,1H),5.00(dd,J=8.0,5.0Hz,2H),1.25(s,9H); 13C NMR(101MHz,Chloroform-d)δ194.7,167.2,160.7,158.8,150.0,149.6,148.7,142.1,131.4,131 .2,131.1,130.0,128.6,128.5,127.1,110.4,109.4,90.1,50.5,37.2,36.3,31.4; HRMS(ESI):m / z calcd for C 28 H 29 N 4 O 5 [M+H] + :501.2133found:501.2137.

[0153] (S)-N-(2-((3-(2-(tert-butyl)phenyl)-1-(naphthalen-2-ylmethyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-

[0154] 4-yl)amino)-2-oxoethyl)benzamide(3o)

[0155]

[0156] Yield: 27.47 mg, yield: 49%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR (400MHz, Chloroform-d) δ11.17-10.92(m,1H),7.97(s,1H),7.88-7.83(m,2H),7.83-7.75(m,3H),7.66(ddd,J=14.6,8.3,1.6Hz,2H),7. 53-7.35(m,7H),7.25(d,J=6.0Hz,1H),7.05(dd,J=7.9,1.5Hz,1H),5.32(d,J=5.3Hz,2H),5.13-5.03(m,1H),5.07-4.98(m,2H),1.19(s,9H); 13C NMR(101MHz,Chloroform-d)δ194.6,167.2,161.1,158.7,150.0,148.6,134.7,134.3,133.3,132.9,131.4,131.2,131 .1,131.0,129.2,128.6,128.5,128.1,127.6,127.3,127.1,126.0,125.9,90.2,50.6,44.5,36.3,31.4; HRMS(ESI):m / z calcdfor C 34 H 33 N 4 O 4 [M+H] + :561.2497found:561.2500.

[0157] (S)-N-(2-((1-benzyl-3-(2-iodophenyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)benzamide(3p)

[0158]

[0159] Yield: 28.44 mg, yield: 49%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR(400MHz,Chloroform-d)δ10.98(s,1H),8.01(d,J=8.0Hz,1H),7.81(d,J=7.5Hz,2H),7.51(dt,J=17.6,8.7Hz,4H ),7.45-7.35(m,3H),7.27(dt,J=22.3,7.6Hz,5H),5.23(d,J=10.7Hz,1H),5.21-5.09(m,2H),4.95(d,J=5.0Hz,2H); 13 C NMR(101MHz,Chloroform-d)δ194.6,167.2,160.9,157.3,148.7,141.1,136.8,135.3,134.6,132. 3,131.4,130.7,130.4,129.0,128.5,128.4,127.7,127.1,99.5,90.0,50.6,44.4; HRMS(ESI):m / z calcdfor C 26 H 22IN 4 O 4 [M+H] + :581.0680found:580.0684.

[0160] (S)-N-(2-((1-benzyl-3-(2-(tert-butyl)phenyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)-4-methylbenzamide(3q)

[0161]

[0162] Yield: 24.66 mg, yield: 47%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR (400MHz, Chloroform-d) δ10.94(s,1H),7.66(d,J=7.8Hz,2H),7.59(d,J=8.2Hz,1H),7.42(d,J=7.2Hz,3H),7.29(t,J=7.6Hz,1H),7.17(dq,J =16.6,8.5,7.6Hz,6H),6.97(d,J=7.8Hz,1H),5.12(d,J=13.8Hz,1H),5. 07-4.97(m,2H),4.91(dd,J=12.7,4.8Hz,2H),2.31(s,3H),1.13(s,9H); 13 C NMR(101MHz,Chloroform-d)δ194.7,167.1,161.1,158.6,149.9,148.5,141.7,136.9,131.8,131.08,131.05, 130.88,129.33,129.26,129.1,128.6,128.3,127.6,127.1,90.2,50.5,44.3,36.2,31.4,21.5; HRMS(ESI):m / z calcd forC 31 H 33 N 4 O 4 [M+H] + :525.2497found:525.2495.

[0163] (S)-N-(2-((1-benzyl-3-(2-(tert-butyl)phenyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)-3-methylbenzamide(3r)

[0164]

[0165] Yield: 25.71 mg, Yield rate: 49%, Appearance: white solid; R f (PE / EA = 1:1) = 0.4 - 0.5; 1 1H NMR (400 MHz, Chloroform-d) δ 10.95 (s, 1H), 7.65 - 7.58 (m, 2H), 7.55 (d, J = 6.1 Hz, 1H), 7.43 (dd, J = 7.4, 5.1 Hz, 3H), 7.31 (t, J = 7.5 Hz, 1H), 7.25 - 7.17 (m, 5H), 7.13 (s, 1H), 6.98 (d, J = 7.9 Hz, 1H), 5.13 (d, J = 13.8 Hz, 1H), 5.08 - 4.96 (m, 2H), 4.95 - 4.85 (m, 2H), 2.32 (s, 3H), 1.13 (s, 9H); 13 13C NMR (101 MHz, Chloroform-d) δ 194.7, 167.4, 161.1, 158.6, 149.9, 148.6, 138.3, 136.8, 134.6, 132.1, 131.12, 131.06, 130.9, 129.4, 129.2, 128.6, 128.37, 128.35, 127.9, 127.7, 90.2, 50.5, 44.3, 36.3, 31.4, 21.4; HRMS (ESI): m / z calcd for C 31 H 33 N 4 O 4 [M + H] + : 525.2497 found: 525.2495.

[0166] (S)-N-(2-((1-benzyl-3-(2-(tert-butyl)phenyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)-4-methoxybenzamide(3s)

[0167]

[0168] Yield: 26.49 mg, yield: 49%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1 H NMR(400MHz,Chloroform-d)δ10.95(s,1H),7.73(d,J=8.7Hz,2H),7.60(d,J=8 .1Hz,1H),7.46-7.36(m,3H),7.29(t,J=7.5Hz,1H),7.25-7.13(m,3H),7.08(d, J=5.0Hz,1H),6.97(d,J=7.8Hz,1H),6.84(d,J=8.5Hz,2H),5.12(d,J=13.8Hz, 1H),5.06-4.95(m,2H),4.90(dd,J=12.2,4.9Hz,2H),3.76(s,3H),1.13(s,9H); 13 C NMR(101MHz,Chloroform-d)δ194.8,166.7,162.1,161.1,158.6,149.9,148.6,136.9,131.1,130.9,129. 32,129.27,128.9,128.6,128.3,127.6,127.0,113.7,90.2,55.4,50.5,44.3,36.2,31.4; HRMS(ESI):m / z calcd for C 31 H 33 N 4 O 5 [M+H] + :541.2446found:541.2446.

[0169] (S)-N-(2-((1-benzyl-3-(2-(tert-butyl)phenyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)-4-fluorobenzamide(3t)

[0170]

[0171] Yield: 24.84 mg, yield: 47%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1H NMR(400MHz,Chloroform-d)δ11.02(s,1H),7.85(dd,J=8.5,5.3Hz,2H),7.68(d,J=8.1Hz,1H),7.49(d,J=6.7Hz,3H),7.38(t,J=7.6Hz,1H),7.2 8(p,J=5.4,4.5Hz,3H),7.20(d,J=4.7Hz,1H),7.15-7.00(m,3H),5.18(s ,1H),5.09(d,J=30.2Hz,2H),4.99(dd,J=13.4,4.9Hz,2H),1.21(s,9H); 13 C NMR(101MHz,Chloroform-d)δ194.5,166.1,164.7(d,J=251.2Hz),163.2,161.1,158.7,149.9,148.6,136.8,131.14,13 1.06,131.0,129.5,129.4(d,J=3.9Hz),129.2,128.6,128.4,127.7,115.5(d,J=21.6Hz),90.2,50.5,44.4,36.3,31.4; 19 F NMR(376MHz,Chloroform-d)δ-108.57; HRMS(ESI):m / z calcd for C 30 H 30 FN 4 O 4 [M+H] + :529.2446found:529.2449.

[0172] (S)-N-(2-((1-benzyl-3-(2-(tert-butyl)phenyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)-4-bromobenzamide(3u)

[0173]

[0174] Yield: 25.94 mg, yield: 44%, properties: brown solid; R f (PE / EA=1:1)=0.4-0.5; 1H NMR(400MHz,Chloroform-d)δ10.93(s,1H),7.64(tt,J=5.9,2.0Hz,2H),7.60(d ,J=1.8Hz,1H),7.53-7.46(m,2H),7.42(dt,J=7.7,1.9Hz,3H),7.31(tt,J=7.5, 1.8Hz,1H),7.25-7.13(m,4H),6.98(dt,J=7.9,1.8Hz,1H),5.13(dd,J=13.8,1. 9Hz,1H),5.04(dd,J=13.8,1.9Hz,2H),4.93-4.82(m,2H),1.14(d,J=2.0Hz,9H); 13 C NMR(101MHz,Chloroform-d)δ194.4,166.2,161.1,158.7,149.9,148.6,136.8,133.5,131.7,131.2,131. 05,130.97,129.4,129.2,128.8,128.6,128.4,127.7,126.0,90.2,50.5,44.4,36.3,31.5; HRMS(ESI):m / z calcd for C 30 H 30 Bn 4 O 4 [M+H] + :589.1445found:589.1453.

[0175] (S)-N-(2-((1-benzyl-3-(2-(tert-butyl)phenyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)-3-chlorobenzamide(3v)

[0176]

[0177] Yield: 25.62 mg, yield: 47%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1H NMR(400MHz,Chloroform-d)δ10.93(s,1H),7.76(t,J=1.9Hz,1H),7.68-7.57(m,2H),7.48-7.36(m,4H),7.34-7.26(m,2H) ,7.24-7.12(m,4H),6.98(dd,J=7.8,1.5Hz,1H),5.13(d,J=13.8Hz,1H),5.08-4.97(m,2H),4.96-4.82(m,2H),1.14(s,9H); 13 C NMR(101MHz,Chloroform-d)δ194.3,165.8,161.1,158.7,149.9,148.6,136.8,136.5,134.7,131.4,131.2,131. 05,130.95,129.8,129.4,129.2,128.6,128.4,127.7,127.5,125.2,90.2,50.5,44.4,36.3,31.4; HRMS(ESI):m / z calcd for C 30 H 30 C1N 4 O 4 [M+H] + :545.1950found:545.1954.

[0178] (S)-N-(2-((1-benzyl-3-(2-(tert-butyl)phenyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)-1-naphthamide(3w)

[0179]

[0180] Yield: 19.06 mg, yield: 34%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1H NMR (400MHz, Chloroform-d) δ10.98(s,1H),8.36(d,J=8.2Hz,1H),7.84(d,J=8.3Hz,1H),7.79(d,J=7.8Hz,1H),7.69-7.58(m,2H),7.45( d,J=7.8Hz,6H),7.27-7.12(m,4H),6.98(d,J=7.8Hz,1H),6.92-6.81(m,1H),5.13(s,1H),5.11-5.04(m,2H),4.90(s,2H),1.15(s,9H).; 13 C NMR(101MHz,Chloroform-d)δ194.6,169.5,161.1,160.0,158.7,150.0,148.6,136.9,134.8,133.7,131.2,131.1,131.0, 130.5,129.4,128.6,128.4,128.2,127.7,127.1,126.4,125.8,125.2,124.8,90.3,50.6,44.4,36.3,31.5; HRMS(ESI):m / z calcdfor C 34 H 33 N 4 O 4 [M+H] + :561.2497found:561.2503.

[0181] (S)-N-(2-((1-benzyl-3-(2-(tert-butyl)phenyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)-2-naphthamide(3x)

[0182]

[0183] Yield: 24.67 mg, yield: 44%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1H NMR(400MHz,Chloroform-d)δ11.19-10.85(m,1H),8.38(s,1H),7.90(td,J=16.1,14.9,7.3Hz,4H),7.69(s,1H),7.53 (dd,J=15.3,6.9Hz,4H),7.38(t,J=7.2Hz,2H),7.34-7.23(m,4H),7.06(d,J=7.8Hz,1H),5.22(d,J=13.8Hz,1H),5.16 -5.07(m,2H),5.07-4.97(m,2H),1.21(s,9H); 13 C NMR(101MHz,Chloroform-d)δ194.7,167.2,161.1,158.7,149.9,148.6,136.9,134.8,132.7,131.9,131.14,131.08,131.0, 129.4,129.2,129.0,128.6,128.4,127.8,127.7,127.60,127.55,126.6,123.8,90.3,50.7,44.4,36.3,31.5; HRMS(ESI):m / z calcd for C 34 H 33 N 4 O 4 [M+H] + :561.2497found:561.2502.

[0184] (S)-N-(2-((1-benzyl-3-(2-(tert-butyl)phenyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)thiophene-2-carboxamide(3y)

[0185]

[0186] Yield: 23.25 mg, yield: 45%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1H NMR(400MHz,Chloroform-d)δ11.01(s,1H),7.68(d,J=7.9Hz,1H),7.56(d,J=3.7Hz,1H),7.47(dd,J=15.0,6.1Hz,4H),7.38(t,J=7.6H z,1H),7.32-7.22(m,3H),7.07(td,J=7.8,6.7,3.6Hz,3H),5.20(d,J=13.8Hz,1H),5.14-5.01(m,2H),5.02-4.90(m,2H),1.21(s,9H); 13 C NMR(101MHz,Chloroform-d)δ194.4,161.7,161.1,158.7,149.9,148.6,139.1,136.8,131.13,131.06,13 0.9,129.7,129.3,129.2,128.6,128.4,128.2,127.7,127.6,90.2,50.4,44.4,36.3,31.5; HRMS(ESI):m / z calcd for C 28 H 29 N 4 O 4 S[M+H] + :517.1904 found:517.1906.

[0187] (S)-N-(2-((1-benzyl-3-(2-(tert-butyl)phenyl)-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)amino)-2-oxoethyl)cinnamamide(3z)

[0188]

[0189] Yield: 14.49 mg, yield: 27%, properties: white solid; R f (PE / EA=1:1)=0.4-0.5; 1H NMR(400MHz,Chloroform-d)δ10.95(s,1H),7.67-7.53(m,2H),7.51-7.37(m,5H),7.36-7.25(m,4H),7.24-7.14(m,3H),6.98(dd,J=7.8,1.5H z,1H),6.57(t,J=5.2Hz,1H),6.43(d,J=15.6Hz,1H),5.12(d,J=13.8Hz,1H),5.03(d,J=13.8Hz,2H),4.87(dd,J=9.0,5.1Hz,2H),1.14(s,9H); 13 C NMR(101MHz,Chloroform-d)δ194.6,165.8,161.1,158.6,149.9,148.6,141.0,136.8,135.0,131.12,131.06,13 0.9,129.6,129.4,129.2,128.8,128.6,128.4,127.8,127.7,120.8,90.2,50.3,44.4,36.3,31.5; HRMS(ESI):m / z calcd forC 32 H 33 N 4 O 4 [M+H] + :537.2497 found:537.2493.

[0190] Embodiment 2:

[0191] Chiral 6-NH 2 Uracil compound (S)-1j (having the opposite configuration to the product (R)-1j in Example 1), the reaction formula and specific reaction process are as follows:

[0192]

[0193] Racemic 1j (0.1 mmol, 38.5 mg), resolution reagent azlactone 2a (0.05 mmol, 8.0 mg), chiral phosphoric acid catalyst A (0.005 mmol, 3.8 mg) were placed in a 5 mL reaction tube, and 50.0 mg The mixture was stirred for 3 hours at room temperature with 1 mL of ultra-dry dichloromethane and molecular sieves. After the reaction was complete by thin layer chromatography, silica gel column chromatography was used with petroleum ether and ethyl acetate as eluents to purify the obtained white solid (S)-1j (18.9 mg, 49% yield, 96:4er).

[0194] Embodiment 3:

[0195] Chiral 6-NH 2 Uracil compound (S)-1i (having the opposite configuration to the product (R)-1i in Example 1), the reaction formula and specific reaction process are as follows:

[0196]

[0197] Racemic 1i (0.1 mmol, 38.5 mg), resolution reagent azlactone 2a (0.05 mmol, 8.0 mg), chiral phosphoric acid catalyst A (0.005 mmol, 3.8 mg) were placed in a 5 mL reaction tube, and 50.0 mg The mixture was stirred for 3 hours at room temperature with 1 mL of ultra-dry dichloromethane and molecular sieves. After the reaction was complete by thin layer chromatography, silica gel column chromatography was used with petroleum ether and ethyl acetate as eluents to purify the obtained white solid (S)-1i (19.3 mg, 49% yield, 93:7 er).

[0198] Embodiment 4:

[0199] In this example, racemic uracil 1a and azlactone 2a were used as reaction substrates, and the effects on reaction yield and enantioselectivity were investigated by changing the reaction conditions. The specific process is as follows (i.e., the optimal reaction condition is None):

[0200]

[0201] The reaction conditions were changed based on the above reaction process. The changes in the reaction conditions and their effects on the reaction yield and enantioselectivity are shown in the following table:

[0202]

[0203]

[0204] Note: The yield is the separation yield, and the corresponding selectivity ratio er (enantiomeric ratio) value is measured by HPLC. The selectivity factor s is calculated as follows: s = ln [(1-C)(1-ee sub )] / ln[(1-C)(1+ee sub )],C=ee sub / (ee pro +ee sub );where ee sub To recover the enantiomeric excess of raw material (R)-1, ee pro The enantiomeric excess of product (S)-3.

[0205] It can be seen from the above table that under the reaction conditions of the present invention, the corresponding chiral 6-NH 2 The recovery rate of uracil is as high as 49%, and the enantioselectivity is 99:1. A series of control experiments show that the catalyst (S)-A1, solvent ultra-dry dichloromethane, feed ratio 1a:2a=1:0.5 and additives Molecular sieves have an impact on the reaction. If they are changed, the yield, enantioselectivity and selectivity factor will all decrease.

[0206] Experimental Example 1:

[0207] Example 2 prepared chiral 6-NH 2 The cytotoxicity test of uracil compound (S)-1j on human colon cancer cells, the specific test process is as follows:

[0208] Human colon cancer cells (HCT116) were inoculated in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in DMEM complete medium containing 10% fetal bovine serum, and compound (S)-1j or irinotecan was added at a concentration of 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compound (S)-1j and irinotecan were used as negative controls, and culture medium without cells was used as blanks. At the end of exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0209] IC 50 IC is the half inhibitory concentration, which is the drug concentration that inhibits the growth of half of the cancer cells. 50 The lower the value, the greater the cytotoxic effect. Graphpad Prism 9.0 software was used to calculate the cell survival rate to obtain the IC 50 value.

[0210] Test results such as Figure 1 The results showed that compound (S)-1j had significant cytotoxicity against human colon cancer cell HCT116, and its IC 50 The value is 8.34 μM; the IC of irinotecan 50 The value is 9.14 μM, and both have comparable cytotoxicity.

[0211] Experimental Example 2:

[0212] Example 2 prepared chiral 6-NH 2 The cytotoxicity test of uracil compound (S)-1j on human cervical cancer cells, the specific test process is as follows:

[0213] Human cervical cancer cells (HeLa) were seeded in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in DMEM complete medium containing 10% fetal bovine serum, and compound (S)-1j or irinotecan was added at a concentration of 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compound (S)-1j and irinotecan were used as negative controls, and culture medium without cells was used as blanks. At the end of exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0214] Test results such as Figure 2 The results showed that compound (S)-1j had good cytotoxicity against human cervical cancer cells Hela, and its IC 50 The value is 16.08 μM; the IC of irinotecan 50 The value is 15.41μM, and both have comparable cytotoxicity.

[0215] Experimental Example 3:

[0216] Example 2 prepared chiral 6-NH 2 The cytotoxicity test of uracil compound (S)-1j on human multiple myeloma cells, the specific test process is as follows:

[0217] Human multiple myeloma cells (U266) were seeded in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in RPMI-1640 complete medium containing 10% fetal bovine serum, and compound (S)-1j or irinotecan was added at a concentration of 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compound (S)-1j and irinotecan were used as negative controls, and culture medium without cells was used as blanks. At the end of exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0218] Test results such as Figure 3 The results showed that compound (S)-1j had good cytotoxicity against human multiple myeloma cells U266, and its IC 50 The value is 6.88 μM; the IC of irinotecan50 The value is 8.42μM, and both have comparable cytotoxicity.

[0219] Experimental Example 4:

[0220] Example 2 prepared chiral 6-NH 2 The cytotoxicity test of uracil compound (S)-1j on human prostate cancer cells, the specific test process is as follows:

[0221] Human prostate cancer cells (PC3) were seeded in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in RPMI-1640 complete medium containing 10% fetal bovine serum, and compound (S)-1j or irinotecan was added at a concentration of 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compound (S)-1j and irinotecan were used as negative controls, and culture medium without cells was used as blanks. At the end of exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0222] Test results such as Figure 4 The results showed that compound (S)-1j had good cytotoxicity against human prostate cancer cells PC3, and its IC 50 The value is 17.91 μM; the IC of irinotecan 50 The value is 16.03μM, and both have comparable cytotoxicity.

[0223] Experimental Example 5:

[0224] Example 1 prepared chiral 6-NH 2 The cytotoxicity test of uracil compounds (S)-1j, (R)-1j and rac-1j on human renal cell adenocarcinoma cells ACHN was conducted. The specific test process is as follows:

[0225] Human renal cell adenocarcinoma cells ACHN were seeded in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in RPMI-1640 complete medium containing 10% fetal bovine serum, and compounds (S)-1j, (R)-1j and rac-1j were added at concentrations ranging from 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compounds (S)-1j, (R)-1j and rac-1j were used as negative controls, and culture medium without cells was used as blanks. At the end of exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0226] Test results such as Figure 5 The results showed that compounds (S)-1j, (R)-1j and rac-1j had good cytotoxicity against human renal cell adenocarcinoma cells ACHN.

[0227] Experimental Example 6:

[0228] Example 1 prepared chiral 6-NH 2 The cytotoxicity test of uracil compounds (S)-1j, (R)-1j and rac-1j on human prostate cancer cells LNCaP was conducted. The specific test process is as follows:

[0229] Human prostate cancer cells LNCaP were seeded in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in RPMI-1640 complete medium containing 10% fetal bovine serum, and the compounds (S)-1j, (R)-1j and rac-1j were added at concentrations ranging from 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compounds (S)-1j, (R)-1j and rac-1j were used as negative controls, and the culture medium without cells was used as a blank. At the end of the exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0230] Test results such as Figure 6 The results showed that compounds (S)-1j, (R)-1j and rac-1j had good cytotoxicity against human prostate cancer cells LNCaP.

[0231] Experimental Example 7:

[0232] Example 1 prepared chiral 6-NH 2The cytotoxicity test of uracil compounds (S)-1j, (R)-1j and rac-1j on human liver cancer cells HepG2 was conducted. The specific test process is as follows:

[0233] Human hepatoma cells HepG2 were seeded in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in RPMI-1640 complete medium containing 10% fetal bovine serum, and compounds (S)-1j, (R)-1j and rac-1j were added at concentrations ranging from 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compounds (S)-1j, (R)-1j and rac-1j were used as negative controls, and culture medium without cells was used as blanks. At the end of exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0234] Test results such as Figure 7 The results showed that compounds (S)-1j, (R)-1j and rac-1j had good cytotoxicity against human liver cancer cells HepG2.

[0235] Experimental Example 8:

[0236] Example 1 prepared chiral 6-NH 2 The cytotoxicity test of uracil compounds (S)-1i, (R)-1i and rac-1i on human renal cell adenocarcinoma cells ACHN was conducted. The specific test process is as follows:

[0237] Human renal cell adenocarcinoma cells ACHN were seeded in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in RPMI-1640 complete medium containing 10% fetal bovine serum, and compounds (S)-1i, (R)-1i and rac-1i were added at concentrations ranging from 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compounds (S)-1i, (R)-1i and rac-1i were used as negative controls, and culture medium without cells was used as blanks. At the end of exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0238] Test results such as Figure 8 The results showed that compounds (S)-1i, (R)-1i and rac-1i had good cytotoxicity against human renal cell adenocarcinoma cells ACHN.

[0239] Experimental Example 9:

[0240] Example 1 prepared chiral 6-NH 2 The cytotoxicity test of uracil compounds (S)-1i, (R)-1i and rac-1i on human colon cancer cells HCT116 was conducted. The specific test process is as follows:

[0241] Human colon cancer cells HCT116 were inoculated in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in RPMI-1640 complete medium containing 10% fetal bovine serum, and compounds (S)-1i, (R)-1i and rac-1i were added at concentrations ranging from 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compounds (S)-1i, (R)-1i and rac-1i were used as negative controls, and culture medium without cells was used as blanks. At the end of exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0242] Test results such as Fig. 9 The results showed that compounds (S)-1i, (R)-1i and rac-1i had good cytotoxicity against human colon cancer cells HCT116.

[0243] Experimental Example 10:

[0244] Example 1 prepared chiral 6-NH 2 The cytotoxicity test of uracil compounds (S)-1i, (R)-1i and rac-1i on human cervical cancer cells HeLa is as follows:

[0245] Human cervical cancer cells HeLa were inoculated in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in RPMI-1640 complete medium containing 10% fetal bovine serum, and compounds (S)-1i, (R)-1i and rac-1i were added at concentrations ranging from 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compounds (S)-1i, (R)-1i and rac-1i were used as negative controls, and culture medium without cells was used as blanks. At the end of exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0246] Test results such as Fig.10The results showed that compounds (S)-1i, (R)-1i and rac-1i had good cytotoxicity against human cervical cancer cells HeLa.

[0247] Experimental Example 11:

[0248] Example 1 prepared chiral 6-NH 2 The cytotoxicity test of uracil compounds (S)-1i, (R)-1i and rac-1i on human prostate cancer cells LNCaP is as follows:

[0249] Human prostate cancer cells LNCaP were seeded in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in RPMI-1640 complete medium containing 10% fetal bovine serum, and the compounds (S)-1i, (R)-1i and rac-1i were added at concentrations ranging from 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compounds (S)-1i, (R)-1i and rac-1i were used as negative controls, and culture medium without cells was used as blanks. At the end of exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0250] Test results such as Fig.11 The results showed that compounds (S)-1i, (R)-1i and rac-1i had good cytotoxicity against human prostate cancer cells LNCaP.

[0251] Experimental Example 12:

[0252] Example 1 prepared chiral 6-NH 2 The cytotoxicity test of uracil compounds (S)-1i, (R)-1i and rac-1i on human multiple myeloma cells U266 is as follows:

[0253] Human multiple myeloma cells U266 were seeded in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in RPMI-1640 complete medium containing 10% fetal bovine serum, and compounds (S)-1i, (R)-1i and rac-1i were added at concentrations ranging from 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compounds (S)-1i, (R)-1i and rac-1i were used as negative controls, and culture medium without cells was used as blanks. At the end of exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0254] Test results such as Fig.12 The results showed that compounds (S)-1i, (R)-1i and rac-1i had good cytotoxicity against human multiple myeloma cells U266.

[0255] Experimental Example 13:

[0256] Example 1 prepared chiral 6-NH 2 The cytotoxicity test of uracil compounds (S)-1i, (R)-1i and rac-1i on human prostate cancer cells LNCaP is as follows:

[0257] Human prostate cancer cells LNCaP were seeded in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in RPMI-1640 complete medium containing 10% fetal bovine serum, and the compounds (S)-1i, (R)-1i and rac-1i were added at concentrations ranging from 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compounds (S)-1i, (R)-1i and rac-1i were used as negative controls, and culture medium without cells was used as blanks. At the end of exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0258] Test results such as Fig.13 The results showed that compounds (S)-1i, (R)-1i and rac-1i had good cytotoxicity against human prostate cancer cells LNCaP.

[0259] Experimental Example 14:

[0260] Example 1 prepared chiral 6-NH 2The cytotoxicity test of uracil compounds (S)-1i, (R)-1i and rac-1i on human liver cancer cells HepG2 was conducted. The specific test process is as follows:

[0261] Human hepatoma cells HepG2 were seeded in a 96-well cell culture dish, with 5000 cells per well. The cells were cultured for 24 hours in RPMI-1640 complete medium containing 10% fetal bovine serum, and compounds (S)-1i, (R)-1i and rac-1i were added at concentrations ranging from 0.1 μM to 200 μM, and the cells were cultured for another 48 hours. Cells not exposed to compounds (S)-1i, (R)-1i and rac-1i were used as negative controls, and culture medium without cells was used as blanks. At the end of exposure, CCK-8 reagent was added at 10 μL / well and incubated at 37°C in the dark for 1-3 hours. Then, the culture plate was shaken for 10 seconds, and the optical density value of each well was measured at a wavelength of 450 nm using a microplate reader. N=6 for each experimental group, and the experiment was repeated three times.

[0262] Test results such as Fig.14 The results showed that compounds (S)-1i, (R)-1i and rac-1i had good cytotoxicity against human liver cancer cells HepG2.

[0263] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A chiral 6-NH2 uracil compound, characterized in that: The general structure is as follows: Among them, R 1 is a heterocyclic or substituted heterocyclic ring, an aryl or substituted aryl group; R 2 is a hydrogen atom, an aryl group or a substituted aryl group; R 3 is tert-butyl or halogen.

2. The chiral 6-NH2 uracil compound according to claim 1, characterized in that: R 1 is phenyl, methoxy-substituted phenyl, halogen-substituted phenyl, trifluoromethyl-substituted phenyl, thienyl, furanyl, naphthyl; Hydrogen atom, methoxy group, halogen, trifluoromethyl group, thienyl group, furyl group, phenyl group; R 2 is a hydrogen atom, a phenyl group, a methyl group, a phenyl group substituted with a tert-butyl group, a phenyl group substituted with a methoxy group, or a phenyl group substituted with a halogen group; R 3 is tert-butyl or halogen.

3. The chiral 6-NH2 uracil compound according to claim 2, characterized in that: The specific structural formula of chiral 6-NH2 uracil compounds is as follows:

4. The method for preparing the chiral 6-NH2 uracil compound according to any one of claims 1 to 3, characterized in that: The reaction formula is as follows: Wherein, R is an aryl group or a substituted aryl group; The specific preparation process is as follows: The racemic uracil, azlactone, chiral phosphoric acid catalyst, additive and solvent are mixed, and the mixture is stirred at room temperature for reaction for 36-72 hours. After the reaction is completed, the mixture is separated and recovered to obtain a chiral 6-NH2 uracil compound.

5. The method for preparing chiral 6-NH2 uracil compounds according to claim 4, characterized in that: The molar ratio of racemic uracil, azlactone and chiral phosphoric acid catalyst is 1:0.5-0.6:0.03-0.

06.

6. The method for preparing chiral 6-NH2 uracil compounds according to claim 4, characterized in that: The structural formula of azlactone is as follows:

7. The method for preparing chiral 6-NH2 uracil compounds according to claim 4, characterized in that: The structural formula of the chiral phosphoric acid catalyst is as follows:

8. The method for preparing chiral 6-NH2 uracil compounds according to claim 4, characterized in that: The solvent is dry dichloromethane, chloroform, carbon tetrachloride, 1,2-dichloroethane, ether, tetrahydrofuran, toluene or acetonitrile; the additive is Molecular sieves, Molecular sieves, Molecular sieve or Na2SO4, added amount is 40-60 mg / mmol.

9. Use of the chiral 6-NH2 uracil compound according to any one of claims 1 to 3 in the preparation of anti-tumor drugs.

10. The use according to claim 9, characterized in that: The tumors were renal cell adenocarcinoma, colon cancer, cervical cancer, multiple myeloma, prostate cancer, and liver cancer.

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