Oxygen glycoside bond linked adenosine diphosphate ribose polypeptide as well as synthesis method and application thereof

Abstract

The invention discloses an adenosine diphosphate ribose polypeptide linked by an oxygen glycoside bond as well as a synthesis method and application of the adenosine diphosphate ribose polypeptide. The structural formula of the adenosine diphosphate ribose polypeptide is # imgabs0 #. The invention also specifically discloses a synthesis method of the adenosine diphosphate ribose polypeptide linked by the oxygen glycoside bond and application of the adenosine diphosphate ribose polypeptide as a protein ADP-nuclear saccharification simulant. The method has the advantages of good reaction specificity, high selectivity and simple reaction steps. Wherein the binding constants of the ADP-ribosyl polypeptides T-2-alpha, T-3-alpha, T-3-beta, S-1-alpha and S-2-alpha and the MacroH2A 1.1 protein reach the level of 6.24-9.35 [mu] M, which is close to the binding level of natural ADP-ribosyl polypeptides and the MacroH2A 1.1 protein, which shows that the compounds can be effectively used as the ligand molecules of the MacroH2A 1.1 protein, and can be used for preparing the MacroH2A 1.1 protein. Important design basis and lead compound structures are provided for the development of ADPr-recognition protein specific inhibitors.

Description

Technical Field

[0001] The invention belongs to the technical field of chemical bionic O-ADP-ribosylation reaction and application, and specifically relates to a class of adenosine diphosphate ribose polypeptides linked by oxygen glycosidic bonds, and a synthesis method and application thereof. Background Art

[0002] Adenosine diphosphate ribopolypeptide (ADP-ribopolypeptide) is an important tool for studying protein ADP-ribosylation. It participates in a series of important physiological and pathological processes, such as DNA repair and replication, chromosome structure regulation and transcription, cell differentiation, etc. Oxoside-linked ADP-ribopolypeptide is a class of ADP-ribopolypeptide with important physiological functions; designing and synthesizing such polypeptides is of great significance for studying the physiological function of protein ADP-ribosylation, discovering new drug targets and designing drug lead compounds.

[0003] The traditional synthetic route of ADP-ribosyl compounds usually starts with base-modified nucleosides as starting materials, followed by phosphorylation. This synthetic route requires various nucleotides as reactants to react under harsh reaction conditions. In addition, the various nucleotide intermediates obtained in the reaction have high polarity and high water solubility, which makes the purification and post-processing procedures of these compounds very cumbersome and inefficient. Therefore, it is very important and challenging to develop new concise synthetic routes to effectively synthesize structurally diverse ADP-glycosylated compounds. Summary of the invention

[0004] The technical problem solved by the invention is to provide a class of adenosine diphosphate ribose polypeptides linked by oxygen glycosidic bonds and a synthesis method and application thereof.

[0005] The present invention adopts the following technical solution to solve the above technical problems. A class of adenosine diphosphate ribose polypeptides linked by oxygen glycosidic bonds has the structural formula:

[0006]

[0007] Wherein R is a polypeptide, and the glycosidic bond between the oxygen atom and the ribose is an α-configuration or a β-configuration.

[0008] The method for synthesizing the adenosine diphosphate ribose polypeptide linked by an oxygen glycosidic bond of the present invention comprises the following specific steps: using nicotinamide adenine dinucleotide (NAD + ) and a polypeptide containing a terminal hydroxyl group as raw materials to prepare the target product adenosine diphosphate ribopolypeptide; or nicotinamide adenine dinucleotide (NAD +) and alcohol compounds with functional groups are used as raw materials to prepare α- / β-type ADPr-mimics, and then the α- / β-type ADPr-mimics and a polypeptide containing a terminal azide group or a polypeptide containing a terminal thiol group are used as raw materials to prepare the target product adenosine diphosphate ribose polypeptide; the synthetic route corresponding to the synthetic process is:

[0009]

[0010] The method for synthesizing the adenosine diphosphate ribose polypeptide linked by an oxygen glycosidic bond of the present invention comprises the following specific steps: under anhydrous conditions, nicotinamide adenine dinucleotide β-NAD + The polypeptide containing the terminal hydroxyl group and the hydroxyl donor is added to a mixed solution of ionic liquid [TMG][BF4], tetramethylguanidine (TMG) and DMSO, and the resulting reaction system is stirred at 70-80°C for reaction, and then nicotinamide adenine dinucleotide β-NAD is repeatedly added to the reaction system. + , and then react with stirring at 70-80°C to obtain the target product adenosine diphosphate ribose polypeptide.

[0011] The method for synthesizing the adenosine diphosphate ribose polypeptide linked by an oxygen glycosidic bond of the present invention comprises the following specific steps: under anhydrous conditions, nicotinamide adenine dinucleotide β-NAD + and alcohol compounds are added to the ionic liquid [TMG][BF4], and the reaction system is stirred at 70-80°C to obtain an α- / β-type ADPr-mimetic containing an alkynyl group; a polypeptide containing a terminal azide group, an α- / β-type ADPr-mimetic containing an alkynyl group, and CuSO 4 ·5H 2 O, sodium ascorbate (NaASC), and tris (3-hydroxypropyltriazolemethyl) amine (THPTA) are added to water, and the resulting reaction system is stirred at room temperature to react, and the ADPr group is modified on the polypeptide through a click reaction to obtain the target product adenosine diphosphate ribose polypeptide.

[0012] The method for synthesizing the adenosine diphosphate ribose polypeptide linked by an oxygen glycosidic bond of the present invention comprises the following specific steps: under anhydrous conditions, nicotinamide adenine dinucleotide β-NAD +and alcohol compounds are added to the ionic liquid [TMG][BF4], and the reaction system is stirred at 70-80°C to obtain an α- / β-type ADPr-mimetic containing an alkynyl group; a polypeptide containing a terminal thiol group, dithiothreitol (DTT), an α- / β-type ADPr-mimetic containing an alkynyl group and phenyl (2,4,6-trimethylbenzoyl) lithium phosphate (LAP) are dissolved in water, and the resulting reaction system is placed on ice, and irradiated with 365nm light at a distance of more than 15cm from the sample to react, and the ADPr group is modified on the polypeptide through a thioindene reaction to obtain the target product adenosine diphosphate ribose polypeptide.

[0013] The invention discloses an application of the oxygen-glycosidic bond-linked adenosine diphosphate ribose polypeptide as a protein ADP-ribosylation mimetic.

[0014] The present invention provides a commercial nicotinamide adenine dinucleotide NAD + A new method for synthesizing 1-2 steps of adenosine diphosphate ribose polypeptide linked by an oxygen glycosidic bond using 2-hydroxy-2-nitropropene as raw material under stereochemical control has the following characteristics: 1. Good reaction specificity and high selectivity; 2. Simple reaction steps without complicated protection and deprotection procedures; 3. The structural diversification modification of adenosine diphosphate ribose polypeptide can be easily achieved. DETAILED DESCRIPTION

[0015] The above contents of the present invention are further described in detail below through examples, but this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples, and all technologies implemented based on the above contents of the present invention belong to the scope of the present invention.

[0016] Example 1

[0017]

[0018] Under anhydrous conditions, β-NAD + (0.15 mmol) and a hydroxyl donor (0.015 mmol) were added to a mixed solution of ionic liquid [TMG][BF4] (0.75 mmol), TMG (0.75 mmol) and DMSO (5 mL), and the resulting reaction system was heated at 75 °C with stirring for 3 h, and then β-NAD was repeatedly added to the mixture. + (0.15 mmol), and then heated with stirring at 75°C for 3 hours. Afterwards, the reaction system was extracted with water and dichloromethane, and the aqueous phase was collected. Semi-preparative HPLC (C18, CH 3 CN / 0.1M TEAB=40 / 60 (V / V), flow rate=3.0 mL / min in 40 minutes, I=254 nm) to separate and purify the product. After freeze-drying, a white powder product can be obtained.

[0019] 1 H NMR (600 MHz, D 2 O)δ8.53(s,1H),8.28(s,1H),6.15(d,J=6.0Hz,1H),4.94(d,J=4.8Hz,1H),4.54(t,J=4.8Hz,1H),4.42–4.36(m,2H),4.28(t,J=6.6Hz,1H), 4.24–4.21(m,1H),4.20–4.17(m,1H),4.16–4.14(m,1H),4.13–4.10(m ,1H),4.06–4.00(m,3H),3.95–3.91(m,1H),3.79(s,3H),1.42(s,9H). 31 P NMR (243MHz, D 2 O)δ-11.33(s).MALDI-TOF-MS:m / z C 24 H 38 N 6 O 18 P 2 calculated[MH] - =759.16,found=759.33.

[0020] Example 2

[0021]

[0022] Under anhydrous conditions, β-NAD + (0.15 mmol) and a hydroxyl donor (0.015 mmol) were added to a mixed solution of ionic liquid [TMG][BF4] (0.75 mmol), TMG (0.75 mmol) and DMSO (5 mL), and the resulting reaction system was stirred and heated at 75 °C for 3 hours, and then β-NAD was repeatedly added to the reaction system. + (0.15 mmol), and then heated with stirring at 75°C for 3 hours. Afterwards, the reaction system was extracted with water and dichloromethane, and the aqueous phase was collected. Semi-preparative HPLC (C18, CH 3 CN / 0.1M TEAB=40 / 60 (V / V), flow rate=3.0 mL / min in 40 minutes, I=254 nm) to separate and purify the product. After freeze-drying, a white powder product can be obtained.

[0023] 1 H NMR (600MHz, D 2O)δ8.52(s,1H),8.26(s,1H),7.48–7.38(m,4H),7.26–7.21(m,1H),6.14(d,J=600Hz,1H), 5.04(d,J=3.0Hz,1H),4.57–4.50(m,2H),4.40(s,1H),4.23(s,2H),4.19–4.15(m,1H),4.1 4–4.08(m,2H),4.03–3.99(m,2H),3.98–3.92(m,1H),3.88–3.82(m,1H),3.81–3.73(m,2H) ,3.68–3.54(m,1H),3.37–3.31(m,2H),2.47–2.32(m,1H),2.18–2.00(m,3H),1.42(s,9H). 31 P NMR (243MHz, D 2 O)δ-11.33(s).MALDI-TOF-MS:m / zC 34 H 48 N 8 O 18 P 2 calculated M=918.26, found 918.26.

[0024] Example 3

[0025]

[0026] Under anhydrous conditions, β-NAD + (0.15 mmol) and a hydroxyl donor (0.015 mmol) were added to a mixed solution of ionic liquid [TMG][BF4] (0.75 mmol), TMG (0.75 mmol) and DMSO (5 mL), and the resulting reaction system was heated at 75 °C with stirring for 3 h, and then β-NAD was repeatedly added to the mixture. + (0.15 mmol), and then heated with stirring at 75°C for 3 hours. Afterwards, the reaction system was extracted with water and dichloromethane, and the aqueous phase was collected. Semi-preparative HPLC (C18, CH 3 CN / 0.1M TEAB=40 / 60 (V / V), flow rate=3.0 mL / min in 40 minutes, I=254 nm) to separate and purify the product. After freeze-drying, a white powder product can be obtained.

[0027] 1 H NMR (600MHz, D 2O)δ8.50(s,1H),8.25(s,1H),7.46–7.34(m,4H),7.24–7.19(m,1H),6.12(d,J=5.4Hz,1 H),4.96(s,1H),4.57–4.51(m,12H),4.41–4.36(m,1H),4.36–4.32(m,1H),4.24–4.20(m ,2H),4.19–4.15(m,3H),4.13–4.07(m,2H),4.00–3.95(m,2H),3.83–3.74(m,2H),3.71 –3.64(m,1H),3.27(q,J=7.2Hz,2H),2.39–2.33(m,1H),2.14–2.05(m,3H),1.41(s,9H). 31 P NMR (243MHz, D 2 O)δ-11.28(s).MALDI-TOF-MS:m / z C 36 H 51 N 9 O 19 P 2 calculated M=975.28, found 975.32.

[0028] Example 4

[0029]

[0030] Under anhydrous conditions, β-NAD + (0.15 mmol) and a hydroxyl donor (0.015 mmol) were added to a mixed solution of ionic liquid [TMG][BF4] (0.75 mmol), TMG (0.75 mmol) and DMSO (5 mL), and the resulting reaction system was heated at 75 °C with stirring for 3 h, and then β-NAD was repeatedly added to the mixture. + (0.15 mmol), and then heated with stirring at 75°C for 3 hours. Afterwards, the reaction system was extracted with water and dichloromethane, and the aqueous phase was collected. Semi-preparative HPLC (C18, CH 3 CN / 0.1M TEAB=40 / 60 (V / V), flow rate=3.0 mL / min in 40 minutes, I=254 nm) to separate and purify the product. After freeze-drying, a white powder product can be obtained.

[0031] 1 H NMR (400 MHz, D 2O)δ8.52(s,1H),8.26(s,1H),6.13(s,1H),4.96(s,1H),4.53(s,1H),4.44–4.36(m,3H ),4.34–4.28(m,2H),4.21(s,1H),4.11–4.06(m,1H),4.03–3.97(m,2H),3.89–3.82(m ,2H),3.67–3.60(m,2H),3.52–3.45(m,1H),3.35–3.30(m,2H),2.11(s,3H),2.00–1.9 5(m,2H),1.86–1.72(m,3H),1.63–1.56(m,2H),1.42–1.36(m,6H),1.16–1.10(m,2H). 31 P NMR (243MHz, D 2 O)δ-11.28(s).MALDI-TOF-MS:m / zC 36 H 54 F 3 N 11 O 20 P 2 calculated M=1079.30, found 1079.31.

[0032] Example 5

[0033]

[0034] Under anhydrous conditions, β-NAD + (0.015 mmol) and different alcohol compounds (0.75 mmol) were added to the ionic liquid [TMG][BF4] (0.75 mmol), and the resulting reaction system was stirred and heated at 75°C for 6 hours. Afterwards, the reaction system was extracted with water and dichloromethane, and the aqueous phase was collected. Semi-preparative HPLC (C18, CH 3 CN / 0.1M TEAB=20 / 80, within 40 minutes, flow rate=3.0 mL / min, I=254 nm) to separate and purify the product. After freeze-drying, a white powder product can be obtained.

[0035]

[0036] 1 H NMR (400 MHz, D 2O)δ8.53(s,1H),8.27(s,1H),6.15(d,J=6.0 Hz,1H),5.01(t,J=2.0 Hz,1H),4.78–4.75(m,1H),4.56–4.52(m,1H),4.42–4.38(m,1H),4.26–4.18(m,3H),4.15–4.09(m,2H),4.05–4.00(m,2H),3.77–3.69(m,1H),3.63–3.53(m,1H),3.01(q,J=7.2 Hz,1H),2.47(t,J=6.4 Hz,2H). 31 P NMR(162 MHz,D 2 O).δ-11.34(s).MALDI-TOF-MS:m / z C 19 H 26 N 5 O 14 P 2 - ,calculated[M-H] - :610.10,found:610.97。

[0037]

[0038] 1 H NMR(400 MHz,D 2 O)δ8.53(s,1H),8.27(s,1H),6.14(d,J=6.0 Hz,1H),5.00(s,1H),4.77–4.60(m,1H),4.54(t,J=3.6 Hz,1H),4.40(s,1H),4.29–4.21(m,3H),4.15–4.08(m,2H),4.08–3.96(m,2H),3.79–3.71(m,1H),3.57–3.48(m,1H),3.27(q,J=7.2 Hz,1H),2.42(t,J=6.4 Hz,2H). 31 P NMR(162 MHz,D 2 O).δ-11.34(s).MALDI-TOF-MS:m / zC 19 H 26 N 5 O 14 P 2 - ,calculated[M-H] - :610.10,found:610.98。

[0039]

[0040] 1 H NMR(400 MHz,D 2 O)δ8.53(s,1H),8.26(s,1H),6.14(d,J=6.0 Hz,1H),4.94(d,J=4.0 Hz,1H),4.77–4.75(m,1H),4.53(t,J=4.4 Hz,1H),4.39(s,1H),4.26–4.15(m,3H),4.14–4.08(m,2H),4.05–3.96(m,2H),3.77–3.69(m,1H),3.56–3.47(m,1H),3.30–3.15(m,1H),2.24(t,J=7.2 Hz,2H),1.74(t,J=6.8 Hz,2H). 31 P NMR(162 MHz,D 2 O)δ-11.26(s).MALDI-TOF-MS:m / z C 20 H 27 N 5 O 14 P 2 ,calculated M:625.11,found:625.99。

[0041]

[0042] 1 H NMR(400 MHz,D 2 O)δ8.53(s,1H),8.26(s,1H),6.14(d,J=6.0 Hz,1H),4.96(s,1H),4.79–4.76(m,1H),4.54(t,J=4.0 Hz,1H),4.40(s,1H),4.27–4.21(m,3H),4.15–4.08(m,2H),4.05–3.96(m,2H),3.77–3.69(m,1H),3.54–3.47(m,1H),3.30–3.15(m,1H),2.20(t,J=7.2 Hz,2H),1.69(t,J=6.8 Hz,2H). 31 P NMR(162 MHz,D 2 O)δ-11.32(s).MALDI-TOF-MS:m / z C 20 H 27 N 5 O 14 P 2,calculated M:625.11,found:626.07。

[0043]

[0044] 1 H NMR(400 MHz,D 2 O)δ8.54(s,1H),8.27(s,1H),6.15(d,J=6.0 Hz,1H),4.93(d,J=4.0 Hz,1H),4.76–4.73(m,1H),4.56–4.51(m,1H),4.43–4.38(m,1H),4.27–4.21(m,2H),4.18–4.09(m,3H),4.05–3.99(m,2H),3.67–3.58(m,1H),3.47–3.38(m,1H),3.12–3.10(m,1H),2.19(t,J=6.4 Hz,2H),1.66–1.57(m,2H),1.52–1.44(m,2H). 31 P NMR(162 MHz,D 2 O)δ-11.24(s).MALDI-TOF-MS:m / z C 21 H 31 N 5 O 14 P 2 ,calculated M:639.13,found:639.06。

[0045]

[0046] 1 H NMR(400 MHz,D 2 O)δ8.53(s,1H),8.26(s,1H),6.14(d,J=6.0 Hz,1H),4.94(s,1H),4.78–4.75(m,1H),4.54(t,J=7.2 Hz,1H),4.40(s,1H),4.27–4.20(m,3H),4.14–4.07(m,2H),4.03–3.93(m,2H),3.67–3.60(m,1H),3.46–3.68(m,1H),3.11–3.08(m,1H),2.15(t,J=6.4Hz,2H),1.60–1.52(m,2H),1.48–1.41(m,2H). 31 PNMR(162MHz,D 2O)δ-11.31(s).MALDI-TOF-MS:m / z C 21 H 31 N 5 O 14 P 2 , calculated M: 639.13, found: 640.08.

[0047] Example 6

[0048]

[0049] The peptide containing a terminal azide group (0.025 mmol), α- / β-type ADPr-mimetic containing an alkynyl group (0.05 mmol), CuSO 4 ·5H 2 O (0.05 mmol), NaASC (0.05 mmol) and THPTA (0.01 mmol) were added to 500 μL of water, and the resulting mixture was stirred at room temperature for 5 hours to modify the ADPr group on the peptide by click reaction. After the reaction was completed, the ADPr group was modified on the peptide using semi-preparative HPLC (C18, CH 3 CN and 1‰ TFA / H 2 O, 1‰ TFA=30 / 70, within 20 min, flow rate=3.0 mL / min, I=220 nm) to separate and purify the product, and use a vacuum freeze dryer to remove the solvent to obtain a white powdery compound.

[0050]

[0051] MALDI-TOF-MS: m / z C 106 H 180 N 34 O 38 P 2 calculated M=2600.27, found 2600.24.

[0052]

[0053] MALDI-TOF-MS: m / z C 106 H 180 N 34 O 38 P 2 calculated M=2600.27, found 2600.21.

[0054]

[0055] MALDI-TOF-MS: m / z C107 H 182 N 34 O 38 P 2 calculated M=2614.29,found 2614.32。

[0056]

[0057] MALDI-TOF-MS:m / z C 107 H 182 N 34 O 38 P 2 calculated M=2614.29,found 2614.21。

[0058]

[0059] MALDI-TOF-MS:m / z C 108 H 184 N 34 O 38 P 2 calculated M=2628.30,found 2628.29。

[0060]

[0061] MALDI-TOF-MS:m / z C 108 H 184 N 34 O 38 P 2 calculated M=2628.30,found 2628.27。

[0062]

[0063] MALDI-TOF-MS:m / z C 109 H 186 N 34 O 38 P 2 calculated M=2642.32,found 2642.25。

[0064]

[0065] MALDI-TOF-MS:m / z C 109 H 186 N 34 O 38 P2 calculated M=2642.32, found 2642.31.

[0066] Example 7

[0067]

[0068] Under anhydrous conditions, β-NAD + (0.015 mmol) and different alcohol compounds (0.75 mmol) were added to the ionic liquid [TMG][BF4] (0.75 mmol), and the resulting reaction system was stirred and heated at 75°C for 6 hours. Afterwards, the reaction system was extracted with water and dichloromethane, and the aqueous phase was collected. Semi-preparative HPLC (C18, CH 3 CN / 0.1M TEAB=20 / 80 (V / V, within 40 min, flow rate=3.0 mL / min, I=254 nm) to separate and purify the product.

[0069] After freeze-drying, a white powder product can be obtained.

[0070]

[0071] 1 H NMR (400 MHz, D 2 O)δ8.50(s,1H),8.20(s,1H),6.10(d,J=6.0 Hz,1H),5.94–5.80(m,1H),5.30–5.15(m,2H),4.98(d,J=3.6 Hz,1H),4.74(t,J=5.6 Hz,1H),4.51(t,J=4.0 Hz,1H),4.37(s,1H),4.22–4.10(m,6H),4.02–3.97(m,3H). 31 P NMR (162 MHz, D 2 O)δ-11.34(s).MALDI-TOF-MS:m / z C 18 H 26 N 5 O 14 P 2 - ,calculated[MH] - :598.10,found:598.23.

[0072]

[0073] 1 H NMR (400 MHz, D 2O)δ8.50(s,1H),8.21(s,1H),6.11(d,J=6.0 Hz,1H),5.92–5.78(m,1H),5.30-5.16(m,2H),4.98(s,1H),4.77–4.74(m,1H),4.53(t,J=4.0 Hz,1H),4.39(s,1H),4.28-4.20(m,3H),4.16-4.07(m,3H),4.04–3.94(m,3H). 31 P NMR(162 MHz,D 2 O)δ-11.34(s).MALDI-TOF-MS:m / z C 18 H 26 N 5 O 14 P 2 - ,calculated[M-H] - :598.10,found:598.23。

[0074]

[0075] 1 H NMR(400 MHz,D 2 O)δ8.53(s,1H),8.27(s,1H),6.15(d,J=4.4 Hz,1H),5.86–5.74(m,1H),5.15–5.02(m,2H),4.96(s,1H),4.76(t,J=5.6 Hz,1H),4.54(s,1H),4.40(s,1H),4.26–4.20(m,2H),4.19–4.15(m,1H),4.14–4.08(m,2H),4.05–3.98(m,2H),3.71–3.63(m,1H),3.51–3.46(m,1H),2.34–2.25(m,2H). 31 P NMR(162 MHz,D 2 O)δ-11.27(s).MALDI-TOF-MS:m / z C 19 H 28 N 5 O 14 P 2 - calculated[M-H] - =612.11,found 612.15。

[0076]

[0077] 1 H NMR(400 MHz,D 2 O)δ8.52(s,1H),8.26(s,1H),6.14(d,J=6.0 Hz,1H),5.84–5.69(m,1H),5.10–5.00(m,2H),4.96(s,1H),4.84(s,1H),4.54(t,J=3.6 Hz,1H),4.39(s,1H),4.23(s,3H),4.14–4.06(m,2H),4.04–3.95(m,2H),3.74–3.66(m,1H),3.51–3.45(m,1H),2.28–2.21(m,2H). 31 P NMR(162 MHz,D 2 O)δ-11.30(s).MALDI-TOF-MS:m / zC 19 H 28 N 5 O 14 P 2 - calculated[M-H] - =612.11,found 612.22。

[0078]

[0079] 1 H NMR(600 MHz,D 2 O)δ8.52(s,1H),8.26(s,1H),6.14(d,J=5.4 Hz,1H),5.90–5.78(m,1H),5.06–4.94(m,2H),4.93–4.90(m,1H),4.75(t,J=6.0 Hz,1H),4.55–4.50(m,1H),4.41–4.38(m,1H),4.24–4.20(m,2H),4.18–4.14(m,1H),4.12–4.07(m,2H),4.01(t,J=4.2 Hz,2H),3.65–3.59(m,1H),3.43–3.38(m,1H),2.04(q,J=7.2 Hz,2H),1.61(qu,J=7.2 Hz,2H). 31 P NMR(243 MHz,D 2 O)δ-11.17(s).MALDI-TOF-MS:m / zC 20 H 32 N 5 O 14 P 2calculated M=627.13,found 627.13。

[0080]

[0081] 1 H NMR(600 MHz,D 2 O)δ8.51(s,1H),8.25(s,1H),6.12(d,J=6.0 Hz,1H),5.85–5.73(m,1H),5.03–4.95(m,1H),4.92(s,1H),4.78–4.74(m,1H),4.55–4.51(m,1H),4.38(s,1H),4.25–4.20(m,3H),4.13–4.06(m,2H),4.03–3.93(m,2H),3.65–3.59(m,1H),3.43–3.38(m,1H),1.98(q,J=7.2 Hz,2H),1.54(qu,J=7.2 Hz,2H). 31 P NMR(243 MHz,D 2 O)δ-11.23(s).MALDI-TOF-MS:m / z C 20 H 32 N 5 O 14 P 2 calculated M=627.13,found627.13。

[0082]

[0083] 1 H NMR(600MHz,D 2 O)δ8.59(s,1H),8.33(s,1H),6.21(d,J=8.4Hz,1H),5.98–5.87(m,1H),5.14–5.02(m,2H),5.01–4.91(m,1H),4.81(t,J=7.8Hz,1H),4.62–4.57(m,1H),4.46(s,1H),4.29(s,1H),4.24–4.15(m,3H),4.12–4.06(m,2H),3.70–3.60(m,1H),3.50–3.40(m,1H),2.07(q,J=10.2Hz,2H),1.57(qu,J=11.4Hz,2H),1.39(qu,J=12.6Hz,2H). 31 P NMR(243MHz,D 2O)δ-11.27(s).MALDI-TOF-MS:m / z C 21 H 34 N 5 O 14 P 2 calculated M=641.14, found 641.15.

[0084]

[0085] 1 H NMR (600MHz, D 2 O)δ8.59(s,1H),8.33(s,1H),6.21(d,J=8.4Hz,1H),5.93–5.82(m,1H),5.10 –4.98(m,3H),4.86–4.82(m,1H),4.64–4.59(m,1H),4.46(s,1H),4.29(s,3H ),4.22–4.12(m,2H),4.10–4.00(m,2H),3.72–3.64(m,1H),3.52–3.42(m,1H ), 2.07 (q, J = 10.2Hz, 2H), 1.51 (qu, J = 11.4Hz, 2H), 1.36 (qu, J = 12.6Hz, 2H). 31 P NMR (243MHz, D 2 O)δ-11.23.MALDI-TOF-MS:m / z C 21 H 34 N 5 O 14 P 2 calculated M=641.14, found641.16.

[0086] Example 8

[0087]

[0088] The peptide containing a terminal thiol group (0.025 mmol), DTT (0.05 mmol), an α- / β-type ADPr-mimetic containing an alkynyl group (0.05 mmol) and LAP (0.05 mmol) were dissolved in 200 μL of water. The resulting reaction system was placed on ice and irradiated with 365 nm light for 5 minutes from a distance of more than 15 cm from the sample to modify the ADPr group on the peptide through the thionaphthene reaction. After the reaction was completed, the ADPr group was modified on the peptide using a semi-preparative HPLC (C18, CH 3 CN and 1‰ TFA / H 2The product was separated and purified by evaporation of 4% paraformaldehyde (2% ethanol, 1‰ TFA = 30 / 70 (V / V), within 20 min, flow rate = 3.0 mL / min, I = 220 nm), and the solvent was removed using a vacuum freeze dryer to obtain a white powdery compound.

[0089]

[0090] MALDI-TOF-MS: m / z C 106 H 183 N 31 O 38 P 2 S calculated M=2593.26, found 2593.26.

[0091]

[0092] MALDI-TOF-MS: m / z C 106 H 183 N 31 O 38 P 2 S calculated M=2593.26, found 2593.29.

[0093]

[0094] MALDI-TOF-MS: m / z C 107 H 185 N 31 O 38 P 2 S calculated M=2607.27, found 2607.30.

[0095]

[0096] MALDI-TOF-MS: m / z C 107 H 185 N 31 O 38 P 2 S calculated M=2607.27, found 2607.28.

[0097]

[0098] MALDI-TOF-MS: m / z C 108 H 187 N 31 O 38 P 2S calculated M=2621.29, found 2621.23.

[0099]

[0100] MALDI-TOF-MS: m / z C 108 H 187 N 31 O 38 P 2 S calculated M=2621.29, found 2621.33.

[0101]

[0102] MALDI-TOF-MS: m / z C 109 H 189 N 31 O 38 P 2 S calculated M=2635.30, found 2635.34.

[0103]

[0104] MALDI-TOF-MS: m / z C 109 H 189 N 31 O 38 P 2 S calculated M=2635.30, found 2635.28.

[0105] Example 9

[0106] The equilibrium dissociation constants of MacroH2A 1.1 and a total of 16 ADPr-peptides in Examples 6 and 8 were detected by a reichert2 SPR instrument to explore the effects of different linker lengths and configurations on the structure-activity relationship of two ADP-ribosylation-linked polypeptides. MacroH2A 1.1 is an ADPr-recognition protein binding domain present in a variety of species including viruses. The study found that the binding constants of the new structural polypeptides T-2-α, T-3-α, T-3-β, S-1-α, S-2-α designed and synthesized in this patent with MacroH2A 1.1 protein reached 6.24-9.35 μM, which is close to the binding level of natural ADP-ribosyl polypeptides with MacroH2A 1.1 protein, indicating that these compounds can effectively serve as ligand molecules for MacroH2A 1.1 protein, providing important design basis and lead compound structure for the development of ADPr-recognition protein specific inhibitors.

[0107] The binding constants of the 8 polypeptides in Example 6 with MacroH2A 1.1 are:

[0108]

[0109]

[0110] The binding constants of the 8 polypeptides in Example 8 with MacroH2A 1.1 are:

[0111]

[0112] The above embodiments describe the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The above embodiments and descriptions are only for illustrating the principles of the present invention. Without departing from the scope of the principles of the present invention, the present invention may have various changes and improvements, and these changes and improvements all fall within the scope of protection of the present invention.

Claims

1. A class of adenosine diphosphate ribose polypeptides linked by oxygen glycosidic bonds, characterized in that Its structural formula is: Wherein R is a polypeptide, and the glycosidic bond between the oxygen atom and the ribose is an α-configuration or a β-configuration.

2. A method for synthesizing the oxidoglycosidic bond-linked adenosine diphosphate ribose polypeptide according to claim 1, characterized in that The specific process is: using nicotinamide adenine dinucleotide and a polypeptide containing a terminal hydroxyl group as raw materials to prepare the target product adenosine diphosphate ribopolypeptide; or using nicotinamide adenine dinucleotide and an alcohol compound with a functional group as raw materials to prepare an α- / β-type ADPr-mimetic, and then using an α- / β-type ADPr-mimetic and a polypeptide containing a terminal azide group or a polypeptide containing a terminal thiol group as raw materials to prepare the target product adenosine diphosphate ribopolypeptide; the corresponding synthetic route of the synthetic process is:

3. The method for synthesizing the oxidoglycosidic bond-linked adenosine diphosphate ribose polypeptide according to claim 2, characterized in that The specific steps are as follows: under anhydrous conditions, nicotinamide adenine dinucleotide β-NAD + The polypeptide containing the terminal hydroxyl group and the hydroxyl donor is added to a mixed solution of ionic liquid [TMG][BF4], tetramethylguanidine and DMSO, and the resulting reaction system is stirred at 70-80°C for reaction, and then nicotinamide adenine dinucleotide β-NAD is repeatedly added to the reaction system. + , and then react with stirring at 70-80°C to obtain the target product adenosine diphosphate ribose polypeptide.

4. The method for synthesizing the oxidoglycosidic bond-linked adenosine diphosphate ribose polypeptide according to claim 2, characterized in that The specific steps are as follows: under anhydrous conditions, nicotinamide adenine dinucleotide β-NAD + and alcohol compounds are added to the ionic liquid [TMG][BF4], and the reaction system is stirred at 70-80°C to obtain an α- / β-type ADPr-mimetic containing an alkynyl group; a polypeptide containing a terminal azide group, an α- / β-type ADPr-mimetic containing an alkynyl group, CuSO4·5H2O, sodium ascorbate, and tris(3-hydroxypropyltriazolemethyl)amine are added to water, and the obtained reaction system is stirred at room temperature to modify the ADPr group on the polypeptide through a click reaction to obtain the target product adenosine diphosphate ribose polypeptide.

5. The method for synthesizing the ATP-ribose polypeptide linked by an oxygen glycosidic bond according to claim 2, characterized in that The specific steps are as follows: under anhydrous conditions, nicotinamide adenine dinucleotide β-NAD + and alcohol compounds are added to the ionic liquid [TMG][BF4], and the reaction system is stirred at 70-80°C to obtain an α- / β-type ADPr-mimetic containing an alkynyl group; a polypeptide containing a terminal thiol group, dithiothreitol, an α- / β-type ADPr-mimetic containing an alkynyl group and phenyl (2,4,6-trimethylbenzoyl) lithium phosphate are dissolved in water, and the resulting reaction system is placed on ice, and irradiated with 365nm light at a distance of more than 15cm from the sample to react, and the ADPr group is modified on the polypeptide through a thioindene reaction to obtain the target product adenosine diphosphate ribose polypeptide.

6. Use of the oxidoglycosidic bond-linked adenosine diphosphate ribose polypeptide of claim 1 as a protein ADP-ribosylation mimetic.

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