Polypeptide side chain modification method and polypeptide product

Abstract

The invention relates to a polypeptide side chain modification method and a polypeptide product, and the polypeptide side chain modification method comprises the following steps: S001, adding polypeptide, an aryl iodine compound, Cu I, Ni (py) 4Cl2, 2, 2 '-bipiperazine and K2CO3 into a dry threaded sealing pipe; s002, adding an organic solvent for dissolving; s003, performing constant-temperature heating at 100 DEG C in a nitrogen atmosphere for 10 hours; s004, a NaCl solution is added for a quenching reaction; s005, adding an organic extraction liquid for extraction to obtain an extraction liquid; and S006, enabling the extract to flow through a short chromatographic column filled with anhydrous sodium sulfate, carrying out rotary evaporation to remove the solvent, and purifying by utilizing column chromatography to obtain the required polypeptide product. According to the technical scheme, copper is used as a catalyst, and the copper is low in price and biotoxicity. Besides, in the synthetic route of the method, a functional group (such as a fluorine-containing group) can be preset in the aryl compound, so that further conversion of the target compound is facilitated, and the application range of the product is expanded.

Description

Technical Field

[0001] The present invention relates to the technical field of polypeptide modification, and in particular to a method for modifying polypeptide side chains and a polypeptide product. Background Art

[0002] Peptide side chain modification is widely used in the biotechnology industry, mainly using precious metals (palladium, platinum, nickel) or special aromatic sources (triaryl bismuth). Not only is the cost high, but metals such as palladium and platinum also have certain biological toxicity;

[0003] Based on this, the inventors proposed a method for modifying polypeptide side chains that can reduce costs and biological toxicity. Summary of the invention

[0004] In view of this, the purpose of the present invention is to provide a method for modifying the side chain of a polypeptide to solve the problem that the prior art uses precious metals (palladium, platinum, nickel) or special aromatic sources (triaryl bismuth), which are not only costly but also have certain biological toxicity.

[0005] The present invention is achieved through the following technical solutions:

[0006] A method for modifying a polypeptide side chain comprises the following steps:

[0007] S001. Add peptide, aryl iodide compound, CuI, Ni(py)4Cl2, 2,2'-bipiperazine and K2CO into a dry screw-sealed tube. 3；

[0008] S002, adding an organic solvent to dissolve;

[0009] S003, heating at 100°C for 10 h under nitrogen atmosphere;

[0010] S004, adding NaCl solution to quench the reaction;

[0011] S005, adding an organic extract to perform extraction to obtain an extract;

[0012] S006. The extract is passed through a short chromatography column filled with anhydrous sodium sulfate, the solvent is removed by rotary evaporation, and then the desired polypeptide product is purified by column chromatography.

[0013] It is further defined that in said S001:

[0014] The dosage of the peptide is: 0.05-0.1mmol, 1.0equiv;

[0015] The amount of aryl iodide compound used is: 0.25mmol, 2.5equiv;

[0016] The amount of CuI used is: 5.7 mg, 0.03 mmol, 30 mol%;

[0017] The amount of Ni(py)4Cl2 used is: 4.4 mg, 0.01 mmol, 10 mol%;

[0018] The dosage of 2,2'-bipiperazine is: 3 mg, 0.02 mmol, 20 mol%;

[0019] The dosage of K2CO3 is: 55.2 mg, 0.4 mmol, 4.0 equ iv.

[0020] It is further defined that the organic solvent in S002 is a mixed solvent of 1,2-dichloroethane and acetonitrile, wherein the ratio and amount of 1,2-dichloroethane and acetonitrile are respectively: volume ratio 1:1, totaling 1.0 mL.

[0021] It is further defined that the organic extractant in S005 is ethyl acetate.

[0022] A polypeptide product is prepared by the above polypeptide side chain modification method.

[0023] The beneficial effects of the present invention are:

[0024] The method for modifying the side chain of a polypeptide uses copper as a catalyst, which is not only cheap but also has low biological toxicity. In addition, in the synthesis route of the method, a functional group (such as a fluorine-containing group) can be pre-set in the aromatic compound, which is conducive to the further transformation of the target compound and expands the application range of the product.

[0025] Other advantages, objectives and features of the present invention will be described in the following description to some extent, and to some extent, will be obvious to those skilled in the art based on the following examination and study, or can be taught from the practice of the present invention. The objectives and other advantages of the present invention can be realized and obtained through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 For the polypeptide product of Example 1 1 H NMR detection spectrum;

[0027] Figure 2 For the polypeptide product of Example 1 13 C NMR detection spectrum;

[0028] Figure 3 For the polypeptide product of Example 1 19 F NMR detection spectrum;

[0029] Figure 4 For the polypeptide product of Example 21 H NMR detection spectrum;

[0030] Figure 5 For the polypeptide product of Example 2 13 C NMR detection spectrum;

[0031] Figure 6 For the polypeptide product of Example 2 19 F NMR detection spectrum;

[0032] Figure 7 For the polypeptide product of Example 3 1 H NMR detection spectrum;

[0033] Figure 8 For the polypeptide product of Example 3 13 C NMR detection spectrum;

[0034] Fig. 9 For the polypeptide product of Example 3 19 F NMR detection spectrum;

[0035] Fig.10 For the polypeptide product of Example 4 1 H NMR detection spectrum;

[0036] Fig.11 For the polypeptide product of Example 4 13 C NMR detection spectrum;

[0037] Fig.12 For the polypeptide product of Example 4 19 F NMR detection spectrum;

[0038] Fig.13 For the polypeptide product of Example 5 1 H NMR detection spectrum;

[0039] Fig.14 For the polypeptide product of Example 5 13 C NMR detection spectrum;

[0040] Fig.15 For the polypeptide product of Example 5 19 F NMR detection spectrum;

[0041] Fig.16 For the polypeptide product of Example 6 1 H NMR detection spectrum;

[0042] Fig.17 For the polypeptide product of Example 6 13 C NMR detection spectrum;

[0043] Fig.18For the polypeptide product of Example 6 19 F NMR detection spectrum;

[0044] Fig.19 For Example 7 polypeptide product 1 H NMR detection spectrum;

[0045] Fig. 20 For Example 7 polypeptide product 13 C NMR detection spectrum;

[0046] Fig.21 For Example 7 polypeptide product 19 F NMR detection spectrum;

[0047] Fig. 22 For Example 8 polypeptide product 1 H NMR detection spectrum;

[0048] Fig.23 For Example 8 polypeptide product 13 C NMR detection spectrum;

[0049] Fig.24 For Example 8 polypeptide product 19 F NMR detection spectrum; DETAILED DESCRIPTION

[0050] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Generally, the components of the embodiments of the present invention described and shown in the drawings here can be arranged and designed in various different configurations.

[0051] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0052] It should be noted that similar reference numerals and letters denote similar items in the following drawings, and therefore, once an item is defined in one drawing, further definition and explanation thereof is not required in subsequent drawings.

[0053] In the above description of the present invention, it should be noted that the terms "one side", "the other side", etc. indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, or the orientation or positional relationship in which the product of the invention is usually placed when in use, which is only for the convenience of describing the present invention and simplifying the description, and does not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first", "second", etc. are only used to distinguish the description, and cannot be understood as indicating or implying relative importance.

[0054] In addition, the term "same" does not mean that the parts must be absolutely the same, but slight differences are allowed. The term "vertical" only means that the positional relationship between the parts is more vertical than "parallel", and does not mean that the structure must be completely vertical, but can be slightly tilted.

[0055] The present invention provides a technical solution: a method for modifying a polypeptide side chain, comprising the following steps:

[0056] S001. Add peptide, aryl iodide compound, Cu I, Ni(py)4Cl2, 2,2'-bipiperazine and K2CO3 into a dry screw-sealed tube;

[0057] S002, adding an organic solvent to dissolve;

[0058] S003, heating at 100°C for 10 h under nitrogen atmosphere;

[0059] S004, adding NaCl solution to quench the reaction;

[0060] S005, adding an organic extract to perform extraction to obtain an extract;

[0061] S006. The extract is passed through a short chromatography column filled with anhydrous sodium sulfate, the solvent is removed by rotary evaporation, and then the desired polypeptide product is purified by column chromatography.

[0062] In said S001:

[0063] The dosage of the peptide is: 0.05-0.1 mmol, 1.0 equ iv;

[0064] The dosage of the aryl iodide compound is: 0.25 mmol, 2.5 equivalents;

[0065] The dosage of Cu I is: 5.7 mg, 0.03 mmol, 30 mol%;

[0066] The dosage of Ni(py)4Cl2 is: 4.4mg, 0.01mmol, 10mol%;

[0067] The dosage of 2,2'-bipiperazine is: 3 mg, 0.02 mmol, 20 mol%;

[0068] The dosage of K2CO3 is: 55.2 mg, 0.4 mmol, 4.0 equ iv.

[0069] The organic solvent in S002 is a mixed solvent of 1,2-dichloroethane and acetonitrile, wherein the ratio and amount of 1,2-dichloroethane and acetonitrile are respectively: volume ratio 1:1, totaling 1.0 mL.

[0070] The organic extractant in S005 is ethyl acetate.

[0071] Embodiment 1

[0072] Add dipeptide 1 (0.1 mmol, 1.0 equiv.), aryl iodide 2 (0.25 mmol, 2.5 equiv.), Cu I (5.7 mg, 0.03 mmol, 30 mol%), Ni(py)4Cl2 (4.4 mg, 0.01 mmol, 10 mol%), 2,2'-bipiperazine (3 mg, 0.02 mmol, 20 mol%), K2CO3 (55.2 mg, 0.4 mmol, 4.0 equiv.) into a dry 8 mL screw-sealed tube, and then add a mixed solvent of 1,2-dichloroethane and acetonitrile (volume ratio 1:1, total 1.0 mL);

[0073] Heat at 100 °C for 10 h under nitrogen atmosphere, and the progress of the reaction can be monitored by thin layer chromatography;

[0074] The reaction was quenched by adding saline solution, and then extracted with ethyl acetate. The resulting organic solution was passed through a short chromatography column filled with anhydrous sodium sulfate. After the solvent was removed by rotary evaporation, the desired dipeptide product 3 was purified by column chromatography with a yield of 75% as a light yellow solid with a melting point of 52.2-53.6°C.

[0075] Among them, the reaction chemical formula is:

[0076]

[0077] Peptide product detection:

[0078] 1 H NMR (such as Figure 1 shown):

[0079] (400MHz, CDCl3) δ7.90(dd,J=7.7,1.1Hz,1H),7.64(td,J=7.6,1.3Hz,1H),7.57-7.54(m,1H),7.50(td,J=7.2,0.6Hz,1 H),7.42(d,J=7.5Hz,1H),7.14(d,J=4.2Hz,1H),7.13(d,J=4.4Hz,1H),7.08(s,1H),7.05(td,J=5.1,2.0Hz,1H),6.65( d,J=7.4Hz,1H),5.59(s,1H),4.98(s,1H),4.94(dd,J=13.0,5.5Hz,1H),4.15(s,1H),3.71(s,3H),3.39-3.36(m,2H),2 .10-2.06(m,2H),1.37(s,9H),1.35-1.32(m,4H),1.30(d,J=4.8Hz,3H),1.03(s,3H),0.98(s,3H),0.87(t,J=7.1,3H);

[0080] 13 C NMR (such as Figure 2 shown):

[0081] (100MHz, CDCl3) δ200.61 (q, J = 3.9Hz), 172.46, 172.12, 165.30, 155.37, 137.92, 137 .85,132.77,131.16,130.68,128.73,128.32,127.98,127.70,123.26(q,J=272Hz),1 22.74,120.20,118.88,110.52,110.31,104.24,102.23(q,J=33.7Hz),79.99,79.76, 52.92,52.51,50.24,29.40,28.29,27.60,26.71,26.00,25.87,22.21,18.39,13.81;

[0082] 19 F NMR (such as Figure 3 As shown): (376MHz,CDCl3)δ=-64.07(s);

[0083] HRMS-ESI(m / z):[M+Na] + ca lcd.for C 38 H 46 F3N3NaO7 +,736.3180;found,736.3171;

[0084] IR(KBr,cm -1 ):ν3325,2978,1715,1601,1497,1293,742;

[0085] Embodiment 2

[0086] Add dipeptide 4 (0.1 mmol, 1.0 equiv.), aryl iodide 2 (0.25 mmol, 2.5 equiv.), Cu I (5.7 mg, 0.03 mmol, 30 mol%), Ni(py)4Cl2 (4.4 mg, 0.01 mmol, 10 mol%), 2,2'-bipiperazine (3 mg, 0.02 mmol, 20 mol%), K2CO3 (55.2 mg, 0.4 mmol, 4.0 equiv.) into a dry 8 mL screw-sealed tube, and then add a mixed solvent of 1,2-dichloroethane and acetonitrile (volume ratio 1:1, total 1.0 mL);

[0087] Heat at 100 °C for 10 h under nitrogen atmosphere, and the progress of the reaction can be monitored by thin layer chromatography;

[0088] The reaction was quenched by adding saline solution, and then extracted with ethyl acetate. The resulting organic solution was passed through a short chromatography column filled with anhydrous sodium sulfate. After the solvent was removed by rotary evaporation, the desired dipeptide product 5 was purified by column chromatography with a yield of 50% as a light yellow solid with a melting point of 50.4-51.1°C.

[0089] Among them, the reaction chemical formula is:

[0090]

[0091] Peptide product detection:

[0092] 1 H NMR (such as Figure 4 shown):

[0093] (400MHz, CDCl3) δ7.90(d,J=7.6Hz,1H),7.71(s,1H),7.64(t,J=7.6Hz,1H),7.50(t,J=7.6Hz,1H),7 .45(d,J=7.8Hz,1H),7.18-7.13(m,3H),7.10-7.03(m,1H),6.39(s,1H),5.61(s,1H),5.30(s,1H),4. 54(s,1H),3.94(td,J=18.1,5.2Hz,1H),3.86(d,J=17.6Hz,1H),3.66(s,3H),3.47-3.15(m,2H),2.13 -2.04(m,2H),1.44(s,9H),1.39-1.28(m,4H),1.06(s,3H),1.02(s,3H),0.87(td,J=7.0,2.7Hz,3H);

[0094] 13 C NMR (such as Figure 5 shown):

[0095] (100MHz, CDCl3) δ200.64 (q, J = 4.1Hz), 171.90, 169.94, 165.16, 155.56, 138.14, 137.92,132.80,131.24,130.49,128.70,128.19,127.95,123.62(q,J=272Hz),12 2.81,120.31,119.16,111.29,110.17,104.23,102.31(q,J=33.6Hz),80.24,79. 80,52.29,52.28,41.26,29.79,29.42,28.38,26.78,26.73,26.01,22.21,13.79;

[0096] 19 F NMR (such as Figure 6 As shown): (376MHz,CDCl3)δ=-64.08(s);

[0097] HRMS-ESI(m / z):[M+Na] + ca lcd.for C 37 H 44 F3N3NaO7 + ,722.3024;found,722.3038;

[0098] IR(KBr,cm -1):ν3310,2932,1714,1667,1496,1293,1116,741;

[0099] Embodiment 3

[0100] Add dipeptide 6 (0.1 mmol, 1.0 equiv.), aryl iodide 2 (0.25 mmol, 2.5 equiv.), Cu I (5.7 mg, 0.03 mmol, 30 mol%), Ni(py)4Cl2 (4.4 mg, 0.01 mmol, 10 mol%), 2,2'-bipiperazine (3 mg, 0.02 mmol, 20 mol%), K2CO3 (55.2 mg, 0.4 mmol, 4.0 equiv.) into a dry 8 mL screw-sealed tube, and then add a mixed solvent of 1,2-dichloroethane and acetonitrile (volume ratio 1:1, total 1.0 mL);

[0101] Heat at 100 °C for 10 h under nitrogen atmosphere, and the progress of the reaction can be monitored by thin layer chromatography;

[0102] The reaction was quenched by adding saline solution, and then extracted with ethyl acetate. The resulting organic solution was passed through a short chromatography column filled with anhydrous sodium sulfate. After the solvent was removed by rotary evaporation, the desired dipeptide product 7 was purified by column chromatography in a yield of 72% as a white solid with a melting point of 53.2-54.5°C.

[0103] Among them, the reaction chemical formula is:

[0104]

[0105] Peptide product detection:

[0106] 1 H NMR (such as Figure 7 shown):

[0107] (400MHz, CDCl3) δ7.91(dd,J=7.7,1.1Hz,1H),7.70(s,1H),7.61(td,J=7.6,1.1Hz,1H),7.48(t,J= 7.6Hz,1H),7.44(d,J=7.5Hz,1H),7.17-7.11(m,3H),7.08-7.04(m,1H),6.49(s,1H),5.54(s,1H), 5.19(s,1H),4.52(s,1H),4.37(ddd,J=8.3,4.5Hz,1H),3.40-3.22(m,2H),2.11-2.05(m,3H),1.43 (s,9H),1.40(s,9H),1.36-1.27(m,4H),0.94(s,6H),0.86(t,J=7.1Hz,3H),0.83(d,J=6.2Hz,6H);

[0108] 13 C NMR (such as Figure 8 shown):

[0109] (100MHz, CDCl3) δ200.60 (q, J = 3.3Hz), 171.46, 170.60, 165.32, 155.62, 138.08, 137.91 ,132.67,131.34,130.57,128.71,128.28,127.86,127.84,123.64(q,J=273Hz),122.74, 120.23,119.17,111.26,110.20,104.24,102.17(q,J=33.6Hz),81.98,81.97,79.69,57. 66,57.65,31.46,29.40,28.37,28.07,26.76,26.02,25.83,22.21,18.77,17.78,13.79;

[0110] 19 F NMR (such as Fig. 9 As shown): (376MHz,CDCl3)δ=-64.08(s);

[0111] HRMS-ESI(m / z):[M+Na] + ca lcd.for C 43 H 56 F3N3NaO7 + ,806.3963;found,806.3983;

[0112] IR(KBr,cm -1):ν3326,2968,2932,1716,1665,1496,1292,1116,742;

[0113] Embodiment 4

[0114] Add dipeptide 8 (0.1 mmol, 1.0 equiv.), aryl iodide 2 (0.25 mmol, 2.5 equiv.), Cu I (5.7 mg, 0.03 mmol, 30 mol%), Ni(py)4Cl2 (4.4 mg, 0.01 mmol, 10 mol%), 2,2'-bipiperazine (3 mg, 0.02 mmol, 20 mol%), K2CO3 (55.2 mg, 0.4 mmol, 4.0 equiv.) into a dry 8 mL screw-sealed tube, and then add a mixed solvent of 1,2-dichloroethane and acetonitrile (volume ratio 1:1, total 1.0 mL);

[0115] Heat at 100 °C for 10 h under nitrogen atmosphere, and the progress of the reaction can be monitored by thin layer chromatography;

[0116] The reaction was quenched by adding saline solution, and then extracted with ethyl acetate. The resulting organic solution was passed through a short chromatography column filled with anhydrous sodium sulfate. The solvent was removed by rotary evaporation and then purified by column chromatography to obtain the desired dipeptide product 9 in a yield of 72% as a white solid with a melting point of 53.5-54.2°C.

[0117] Among them, the reaction chemical formula is:

[0118]

[0119] Peptide product detection:

[0120] 1 H NMR (such as Fig.10 shown):

[0121] 400MHz, CDCl3) δ7.91(dd,J=7.7,1.1Hz,1H),7.69(s,1H),7.63(td,J=7.7,1.3Hz,1H),7.49(t,J=7.4Hz, 1H),7.44(d,J=7.8Hz,1H),7.17-7.12(m,3H),7.08-7.06(m,1H),6.53(s,1H),5.56(s,1H),5.21(s,1H), 4.49-4.46(m,2H),3.61(s,3H),3.39-3.17(m,2H),2.11-2.03(m,2H),1.78(s,1H),1.44(s,9H),1.37-1. 28(m,6H),0.99(s,3H),0.94(s,3H),0.87(t,J=6.9Hz,3H),0.83(t,J=7.3Hz,3H),0.76(d,J=6.6Hz,3H);

[0122] 13 C NMR (such as Fig.11 shown):

[0123] (100MHz, CDCl3) δ200.61 (q, J = 3.8Hz), 171.83, 171.44, 165.28, 155.62, 138.05, 137.93 ,132.71,131.36,130.57,128.62,128.21,127.91,127.78,123.63(q,J=272Hz),122.77, 120.27,119.14,111.29,110.18,104.22,102.19(q,J=33.6Hz),80.23,79.70,56.65,56. 64,52.04,37.94,29.40,28.36,26.74,26.01,25.83,25.16,22.20,15.27,13.79,11.56;

[0124] 19 F NMR (such as Fig.12 As shown): (376MHz,CDCl3)δ=-64.09(s);

[0125] HRMS-ESI(m / z):[M+Na] + ca lcd.for C 41 H 52 F3N3NaO7 + ,778.3650;found,778.3667;

[0126] IR(KBr,cm -1 ):ν3323,2964,1715,1496,1293,1117,739;

[0127] Embodiment 5

[0128] Add dipeptide 10 (0.1 mmol, 1.0 equiv.), aryl iodide 2 (0.25 mmol, 2.5 equiv.), Cu I (5.7 mg, 0.03 mmol, 30 mol%), Ni(py)4Cl2 (4.4 mg, 0.01 mmol, 10 mol%), 2,2'-bipiperazine (3 mg, 0.02 mmol, 20 mol%), K2CO3 (55.2 mg, 0.4 mmol, 4.0 equiv.) into a dry 8 mL screw-sealed tube, and then add a mixed solvent of 1,2-dichloroethane and acetonitrile (volume ratio 1:1, total 1.0 mL);

[0129] Heat at 100 °C for 10 h under nitrogen atmosphere, and the progress of the reaction can be monitored by thin layer chromatography;

[0130] The reaction was quenched by adding saline solution, and then extracted with ethyl acetate. The resulting organic solution was passed through a short chromatography column filled with anhydrous sodium sulfate. The solvent was removed by rotary evaporation and then purified by column chromatography to obtain the desired dipeptide product 11 in a yield of 75% as a white solid with a melting point of 54.6-56.2°C.

[0131] Among them, the reaction chemical formula is:

[0132]

[0133] Peptide product detection:

[0134] 1 H NMR (such as Fig.13 shown):

[0135] (400MHz, CDCl3)δ7.91(d,J=7.6Hz,1H),7.70(s,1H),7.61(t,J=7.2Hz,1H),7.48t,J=7.5Hz,1H),7 .44(s,1H),7.17-7.11(m,3H),7.09-7.05(m,1H),6.35(s,1H),5.55(s,1H),5.17(s,1H),4.50(s,1 H),4.45(dd,J=13.3,8.0Hz,1H),3.32-3.28(m,2H),2.10-2.15(m,2H),1.64-1.65(m,2H),1.57-1. 50(m,1H)1.43(s,9H),1.40(s,9H),1.35-1.28(m,4H),0.99(s,3H),0.95(m,3H),0.88-0.85(m,9H);

[0136] 13 C NMR (such as Fig.14 shown):

[0137] (100MHz, CDCl3)δ200.60(q,J=4.8Hz),171.75,171.18,165.32,155.58,138.08,137.87,1 32.68,131.33,130.54,128.67,128.34,127.92,127.86,123.63(q,J=272Hz),122.73,120 .25,119.20,111.19,110.19,104.23,102.17(q,J=33.7Hz),81.87,80.12,79.69,51.52,5 1.50,41.98,29.40,28.36,28.01,26.75,26.02,25.83,24.82,22.77,22.21,22.14,13.80;

[0138] 19 F NMR (such as Fig.15 As shown): (376MHz,CDCl3)δ=-64.08(s);

[0139] HRMS-ESI(m / z):[M+Na] + ca lcd.for C 44 H 58 F3N3NaO7 + ,820.4119;found,820.4140;

[0140] IR(KBr,cm-1 ):ν3337,2961,1716,1666,1496,1293,1117,741

[0141] Embodiment 6

[0142] Add tripeptide 30 (0.1 mmol, 1.0 equiv.), aryl iodide 2 (0.25 mmol, 2.5 equiv.), Cu I (5.7 mg, 0.03 mmol, 30 mol%), Ni(py)4Cl2 (4.4 mg, 0.01 mmol, 10 mol%), 2,2'-bipiperazine (3 mg, 0.02 mmol, 20 mol%), K2CO3 (55.2 mg, 0.4 mmol, 4.0 equiv.) into a dry 8 mL screw-sealed tube, and then add a mixed solvent of 1,2-dichloroethane and acetonitrile (volume ratio 1:1, total 1.0 mL);

[0143] Heat at 100 °C for 10 h under nitrogen atmosphere, and the progress of the reaction can be monitored by thin layer chromatography;

[0144] The reaction was quenched by adding saline solution, and then extracted with ethyl acetate. The resulting organic solution was passed through a short chromatography column filled with anhydrous sodium sulfate. After the solvent was removed by rotary evaporation, the desired tripeptide product 31 was purified by column chromatography in a yield of 65% as a light white solid with a melting point of 67.8-69.5°C.

[0145] Among them, the reaction chemical formula is:

[0146]

[0147] Peptide product detection:

[0148] 1 H NMR (such as Fig.16 shown):

[0149] (400 MHz, CDCl3) δ 7.89 (d, J = 7.5 Hz, 1H), 7.62 (t, J = 7.3 Hz, 1H), 7.55 (m, 1H), 7.49 - 7.45 (m, 1H), 7.13 - 7.09 (m, 3H), 7.05 - 7.03 (m, 1H), 6.92 (s, 1H), 6.64 (s, 1H), 5.54 (s, 1H), 5.57 (s, 1H), 4.89 (dd, J = 12.0, 6.6 Hz, 1H), 4.41 (s, 1H), 4.11 - 4.08 (m, 1H), 3.68 (s, 3H), 3.40 - 3.27 (m, 4H), 2.09 - 2.02 (m, 4H), 1.84 - 1.75 (m, 2H), 1.42 (s, 9H), 1.35 - 1.24 (m, 7H), 1.00 (s, 3H), 0.95 (s, 3H), 0.85 (t, J = 6.9 Hz, 3H);

[0150] 13 13C NMR (as Fig.17 shown):

[0151] (100 MHz, CDCl3) δ 200.58 (q, J = 4.2 Hz), 172.12, 171.97, 171.74, 165.32, 155.85, 137.96, 137.85, 132.81, 131.22, 130.58, 128.79, 128.27, 127.89, 127.68, 123.63 (q, J = 273 Hz), 122.70, 120.24, 118.86, 110.66, 110.27, 104.25, 102.21 (q, J = 34.4 Hz), 80.60, 79.72, 59.87, 52.97, 52.47, 48.96, 47.15, 29.39, 28.38, 28.13, 27.60, 26.70, 26.00, 25.82, 24.63, 22.20, 17.58, 13.79;

[0152] 19 19F NMR (as Fig.18 shown): (376 MHz, CDCl3) δ = -64.08 (s);

[0153] HRMS - ESI (m / z): [M + Na] + calcd. for C 43 H 53 F3N4NaO 10 + , 833.3708; found, 833.3731;

[0154] IR(KBr,cm -1 ):ν3306,2933,1693,1498,1292,1118,910,734;

[0155] Embodiment 7

[0156] Add tetrapeptide 59 (0.05 mmol, 1.0 equiv.), aryl iodide 2 (0.25 mmol, 2.5 equiv.), Cu I (5.7 mg, 0.03 mmol, 30 mol%), Ni(py)4Cl2 (4.4 mg, 0.01 mmol, 10 mol%), 2,2'-bipiperazine (3 mg, 0.02 mmol, 20 mol%), K2CO3 (55.2 mg, 0.4 mmol, 4.0 equiv.) into a dry 8 mL screw-sealed tube, and then add a mixed solvent of 1,2-dichloroethane and acetonitrile (volume ratio 1:1, total 1.0 mL);

[0157] Heat at 100 °C for 10 h under nitrogen atmosphere, and the progress of the reaction can be monitored by thin layer chromatography;

[0158] The reaction was quenched by adding saline solution, and then extracted with ethyl acetate. The resulting organic solution was passed through a short chromatography column filled with anhydrous sodium sulfate. The solvent was removed by rotary evaporation and then purified by column chromatography to obtain the desired tetrapeptide product 60 in a yield of 57% as a light yellow solid at 91.0-92.2°C;

[0159] Among them, the reaction chemical formula is:

[0160]

[0161] Peptide product detection:

[0162] 1 H NMR (such as Fig.19 shown):

[0163] (400 MHz, CDCl3) δ 7.91 (d, J = 8.0 Hz, 1H), 7.89 (d, J = 8.0 Hz, 1H), 7.62 - 7.56 (m, 4H), 7.50 - 7.40 (m, 4H), 7.16 - 7.04 (m, 8H), 6.93 (s, 2H), 6.40 (s, 1H), 5.60 - 5.70 (m, 2H), 5.18 - 5.15 (m, 1H), 4.91 (dd, J = 12.0, 5.6 Hz, 1H), 4.39 - 4.37 (m, 1H), 4.30 (m, 1H), 4.26 - 4.24 (m, 1H), 3.71 (s, 3H), 3.45 - 3.36 (m, 2H), 3.27 - 3.17 (m, 2H), 2.10 - 2.05 (m, 4H), 1.38 (s, 9H), 1.36 - 1.26 (m, 14H), 1.12 (s, 6H), 1.00 (s, 6H), 0.86 (t, J = 7.0 Hz, 6H);

[0164] 13 13C NMR (as Fig. 20 shown):

[0165] (100 MHz, CDCl3) δ 200.63 (q, J = 3.4 Hz), 200.61 (q, J = 4.0 Hz), 172.20, 172.19, 172.00, 171.65, 165.40, 165.13, 156.01, 138.05, 137.98, 137.91, 137.80, 132.87, 132.71, 131.36, 131.23, 130.62, 130.44, 128.68, 128.62, 128.40, 128.08, 128.03, 127.85, 127.80, 127.58, 123.64 (q, J = 272 Hz), 123.61 (q, J = 272 Hz), 122.99, 122.65, 120.40, 120.16, 119.05, 118.97, 111.16, 110.68, 110.31, 110.23, 104.29, 104.16, 102.35 (q, J = 34.0 Hz), 102.21 (q, J = 33.2 Hz), 80.81, 79.78, 79.76, 55.58, 53.05, 52.48, 49.55, 49.17, 29.41, 28.29, 27.56, 26.75, 26.01, 25.89, 22.20, 17.70, 13.80;

[0166] 19 19F NMR (as Fig.21As shown): (376MHz,CDCl3)δ=-64.05(s);

[0167] HRMS-ESI(m / z):[M+Na] + ca lcd.for C 70 H 80 F6N6NaO 11 + ,1317.5681; found,1317.5692;

[0168] IR(KBr,cm -1 ):ν3287,2932,1714,1634,1497,1292,1116,738;

[0169] Embodiment 8

[0170] Add pentapeptide 48 (0.05 mmol, 1.0 equiv.), aryl iodide 2 (0.25 mmol, 2.5 equiv.), Cu I (5.7 mg, 0.03 mmol, 30 mol%), Ni(py)4Cl2 (4.4 mg, 0.01 mmol, 10 mol%), 2,2'-bipiperazine (3 mg, 0.02 mmol, 20 mol%), K2CO3 (55.2 mg, 0.4 mmol, 4.0 equiv.) into a dry 8 mL screw-sealed tube, and then add a mixed solvent of 1,2-dichloroethane and acetonitrile (volume ratio 1:1, total 1.0 mL);

[0171] Heat at 100 °C for 10 h under nitrogen atmosphere, and the progress of the reaction can be monitored by thin layer chromatography;

[0172] The reaction was quenched by adding saline solution, and then extracted with ethyl acetate. The resulting organic solution was passed through a short chromatography column filled with anhydrous sodium sulfate. The solvent was removed by rotary evaporation and then purified by column chromatography to obtain the desired tetrapeptide product 49 in a yield of 49% as a white solid with a melting point of 109.2-110.5°C.

[0173] Among them, the reaction chemical formula is:

[0174]

[0175] Peptide product detection:

[0176] 1 H NMR (such as Fig. 22 shown):

[0177] (400MHz, Acetone-D6) δ 7.90 (d, J = 7.8Hz, 1H), 7.84 - 7.70 (m, 4H), 7.64 - 7.56 (m, 3H), 7.23 - 7.10 (m, 9H), 7.03 - 7.00 (m, 1H), 6.30 (d, J = 6.1Hz, 1H), 5.65 - 5.61 (m, 1H), 4.81 (dd, J = 13.1, 7.6Hz, 1H), 4.71 - 4.64 (m, 1H), 4.58 (s, 1H), 4.45 - 4.41 (m, 1H), 4.16 - 4.04 (m, 1H), 3.70 (s, 3H), 3.41 - 3.22 (m, 2H), 3.20 - 3.12 (m, 1H), 3.07 - 2.84 (m, 3H), 2.55 - 2.42 (m, 2H), 2.14 - 2.09 (m, 2H),

[0178] 2.02 (s, 3H), 1.93 - 1.85 (m, 1H), 1.46 - 1.39 (m, 5H), 1.37 (s, 9H), 1.34 - 1.29 (m, 4H), 1.02 (s, 3H), 0.99 (s, 3H), 0.91 (t, J = 7.6Hz, 6H), 0.87 - 0.85 (m, 9H);

[0179] 13 C NMR (as Fig.23 shown):

[0180] (100MHz, Acetone-D6) δ 200.48 (q, J = 3.6Hz), 173.58, 171.70, 171.51, 171.26, 171.10, 165.02, 156.14, 138.61, 138.37, 137.82, 133.54, 131.47, 131.30, 130.00, 129.49, 129.17, 128.83, 128.55, 128.46, 127.03, 124.45 (q, J = 273Hz), 122.92, 120.39, 119.83, 112.72, 110.43, 105.26, 102.23 (q, J = 33.3Hz), 79.59, 79.42, 57.40, 55.70, 54.66, 54.33, 52.99, 51.85, 41.68, 37.76, 37.65, 32.43, 30.40, 28.43, 28.05, 26.85, 26.25, 25.92, 25.64, 25.10, 23.01, 22.48, 22.00, 15.76, 15.01, 13.76, 11.70;

[0181] 19 F NMR (such as Fig.24 As shown): (376MHz,CDCl3)δ=-64.06(s);

[0182] HRMS-ESI(m / z):[M+Na] + ca lcd.for C 61 H 81 F3N6NaO 10 S + ,1169.5579; found,1169.5596;

[0183] IR(KBr,cm -1 ):ν3289,2960,1716,1635,1497,1117,742.

[0184] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solution of the present invention, which should be included in the scope of the claims of the present invention.

Claims

1. A method for modifying a polypeptide side chain, characterized in that: The steps include: S001. Add peptide, aryl iodide compound, CuI, Ni(py)4Cl2, 2,2'-bipiperazine and K2CO3 into a dry screw-sealed tube; S002, adding an organic solvent to dissolve; S003, heating at 100°C for 10 h under nitrogen atmosphere; S004, adding NaCl solution to quench the reaction; S005, adding an organic extract to perform extraction to obtain an extract; S006. The extract is passed through a short chromatography column filled with anhydrous sodium sulfate, the solvent is removed by rotary evaporation, and then the desired polypeptide product is purified by column chromatography.

2. The method for modifying a polypeptide side chain according to claim 1, characterized in that: In said S001: The dosage of the peptide is: 0.05-0.1mmol, 1.0equiv; The amount of aryl iodide compound used is: 0.25mmol, 2.5equiv; The amount of CuI used is: 5.7 mg, 0.03 mmol, 30 mol%; The amount of Ni(py)4Cl2 used is: 4.4 mg, 0.01 mmol, 10 mol%; The amount of 2,2'-bipiperazine used is: 3 mg, 0.02 mmol, 20 mol%; The amount of K2CO3 used is: 55.2 mg, 0.4 mmol, 4.0 equiv.

3. The method for modifying a polypeptide side chain according to claim 2, characterized in that: The organic solvent in S002 is a mixed solvent of 1,2-dichloroethane and acetonitrile, wherein the ratio and amount of 1,2-dichloroethane and acetonitrile are respectively: volume ratio 1:1, totaling 1.0 mL.

4. The method for modifying a polypeptide side chain according to claim 1, characterized in that: The organic extractant in S005 is ethyl acetate.

5. A polypeptide product, characterized in that: The method is prepared by modifying the polypeptide side chain according to any one of claims 1 to 4.

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