Synthetic method of phosphoramidite monomer

Abstract

The invention discloses a synthesis method of a phosphoramidite monomer. The invention specifically discloses a preparation method for preparing a compound shown as a formula III from a compound shown as a formula II. The preparation method has one or more advantages as follows: a new strategy for synthesizing a phosphoramidite monomer with an amino protecting group easy to remove is provided; the reaction steps are few; the cost is low; the overall yield is high; conditions are mild and high in safety; post-treatment is easy; the method is suitable for industrial production; and the phosphoramidite monomer of which the base protecting group is easy to remove can be rapidly prepared in a large scale. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a method for synthesizing phosphoramidite monomers. Background Art

[0002] In the solid-phase synthesis of oligonucleotides (Oligo), the commonly used raw material is phosphoramidite monomer, and the exocyclic amino group of its base usually needs to be protected by a protecting group to reduce the influence on the coupling reaction stage. In the final stage of solid-phase synthesis, the protecting group on the base needs to be removed. Therefore, in order to improve the cycle efficiency of solid-phase synthesis, there are certain requirements for the stability of the selected protecting group.

[0003] In the synthesis process of phosphoramidite monomers, due to the poor stability of the phosphoramidite group, the phosphoramidite group is often introduced in the last step of the synthesis of phosphoramidite monomers, and an easily removable amino protecting group is introduced in the steps before the last step. For example, S.M. Ching et al. reported a method for synthesizing phosphoramidite monomers (the route is shown below), and adopted a synthesis route of introducing an amino protecting group first and finally introducing a phosphoramidite group (Bioorg. Med. Chem. 18 (2010) 6657–6665).

[0004]

[0005] Although more easily removable amino protecting groups will improve the operation efficiency of solid-phase synthesis, relatively speaking, in the process of synthesizing the phosphoramidite monomers protected by them, due to their poor stability and the relatively large number of synthesis steps, side products of protecting group removal are often easily generated in the stages of reaction, post-treatment and purification. Moreover, for some relatively difficult-to-synthesize nucleoside precursors, such as the 2’-OTBS series, the TBS protecting group is extremely likely to migrate between 2’-O and 3’-O, resulting in yield loss (see Becker H F, Corda Y, Mathews M B, et al. [J]. Rna - a Publication of the Rna Society, 1999, 5(7): 865 - 875.). Although US20020150936A1 reported the recovery of 3’-OTBS products, the recovery process requires multiple equilibriums in solvents and additional purification steps, which are time-consuming, costly and have low yields. Therefore, the recovery of 3’-OTBS products is difficult to apply in actual processes, which leads to a significant reduction in the yields of some nucleoside precursor syntheses.

[0006]

[0007] Therefore, the traditional synthetic route to synthesize phosphoramidite monomers with easily removable base protecting groups often has a less than ideal yield and a high cost due to the long reaction route, the easy generation of by-products and the difficulty in purification. In addition, phosphoramidite monomers with easily removable protecting groups have poor stability and are difficult to store for a long time, but de novo synthesis takes a long time, so there are constraints on each other in production, storage and use.

[0008] In view of the importance of synthesizing phosphoramidite monomers with easily removable amino protecting groups, it is urgent to develop a method that has good yield, is easy to operate, has low cost and high safety, is suitable for industrial production and can quickly and massively prepare phosphoramidite monomers with easily removable base protecting groups. Summary of the invention

[0009] In view of the defects of the prior art in synthesizing phosphoramidite monomers with easily removable amino protecting groups, such as long reaction route, easy generation of by-products and difficulty in purification, unsatisfactory yield and high cost, the present invention provides a method for synthesizing phosphoramidite monomers, which has one or more of the following advantages: providing a new strategy for synthesizing phosphoramidite monomers with easily removable amino protecting groups; fewer reaction steps; low cost; high overall yield; mild conditions and high safety; easy post-processing; suitable for industrial production; and capable of rapidly preparing phosphoramidite monomers with easily removable base protecting groups in large quantities.

[0010] The present invention provides a method for preparing compound III, which is the following scheme 1 or scheme 2:

[0011]

[0012] in,

[0013] Base is (*Position directly related to R 1 or Pg2 connected);

[0014] R is H, C1-C4 alkoxy, -OTBS (TBS is tert-butyldimethylsilyl), -OTOM (TOM is (triisopropylsilyloxy)methyl), halogen or -O-MOE (MOE is methoxyethyl);

[0015] Pg2 (The corresponding Base is * position directly connected to Pg2) or (The corresponding Base is * Location directly related to R 1 or Pg2 connected); n is 0, 1 or 2; R 2 is isopropyl or tert-butyl; R X , R Y and RZ Independently H or C 1-6 Alkyl, or, R X And R Z Form ring A with the carbon and nitrogen atoms between them; Ring A is a 4- to 8-membered nitrogen-containing carbocyclic heterocycle;

[0016] Scheme 1: R 1 Is H;

[0017] And when the said Pg2 is It includes the following steps:

[0018] In the presence of an alkyl tertiary amine base, in a halogenated hydrocarbon solvent, the said Compound II and the Pg2 protecting group precursor B are subjected to substitution reaction B to obtain the said Compound III;

[0019] The said Pg2 protecting group precursor B is Where -OR 5 Is a leaving group; n is 0, 1 or 2; R 2 Is isopropyl or tert-butyl;

[0020] Scheme 2: R 1 Is H or C1-C4 alkyl-C(=O)-;

[0021] And when the said Pg2 is It includes the following steps:

[0022] In solvent A, the said Compound II and the Pg2 protecting group precursor A are subjected to substitution reaction A to obtain the said Compound III;

[0023] The said solvent A is an alcohol solvent, a halogenated hydrocarbon solvent or a nitrogen-containing aromatic compound solvent;

[0024] The said Pg2 protecting group precursor A is

[0025] Wherein, R X 、R Y And R Z Independently are H or C 1-6 Alkyl, or, R X And R Z Form ring A with the carbon and nitrogen atoms between them; Ring A is a 4- to 8-membered nitrogen-containing carbocyclic heterocycle.

[0026] In a certain embodiment of the present invention, the said solvent A is an alcohol solvent.

[0027] In a certain embodiment of the present invention, R X 、R Y And R Z Independently are H or C 1-4 Alkyl; The C 1-4The alkyl group is preferably methyl, ethyl or n-butyl.

[0028] In one embodiment of the present invention, R X is H or C 1-4 alkyl.

[0029] In one embodiment of the present invention, R Y and R Z are independently C 1-4 alkyl.

[0030] In one embodiment of the present invention, R X is H or methyl.

[0031] In one embodiment of the present invention, R Y is methyl, ethyl or n-butyl.

[0032] In one embodiment of the present invention, R Z is methyl, ethyl or n-butyl.

[0033] In one embodiment of the present invention, when R X and R Z form ring A with the carbon atom and nitrogen atom between them, Pg2 is For example

[0034] In one embodiment of the present invention, ring A is

[0035] In one embodiment of the present invention, Pg2 is

[0036] In one embodiment of the present invention, is For example

[0037] In one embodiment of the present invention, the is

[0038] The is preferably

[0039] In one embodiment of the present invention, the Pg2 protecting group precursor A is N,N-dimethylformamide dimethyl acetal, N,N-diethylformamide dimethyl acetal, N,N-dimethylacetamide dimethyl acetal, N,N-dibutylformamide dimethyl acetal or 2,2-dimethoxy-1-methylpyrrolidine.

[0040] In one embodiment of the present invention, Pg2 is When, the Pg2 protecting group precursor A is DMF-DMA (i.e., N,N-dimethylformamide dimethyl acetal).

[0041] In a certain embodiment of the present invention, Pg2 is When, the Pg2 protecting group precursor A is DMF-DMA; the solvent A is an alcohol solvent.

[0042] In a certain embodiment of the present invention, Pg2 is When, the Pg2 protecting group precursor A is

[0043] Preferably, the solvent A is a halogenated hydrocarbon solvent.

[0044] In a certain embodiment of the present invention, Pg2 is When, the Pg2 protecting group precursor A is

[0045] Preferably, the solvent A is a nitrogen-containing aromatic compound solvent.

[0046] In a certain embodiment of the present invention, Pg2 is When, the Pg2 protecting group precursor A is

[0047]

[0048] Preferably, the solvent A is a halogenated hydrocarbon solvent.

[0049] In a certain embodiment of the present invention, Pg2 is When, the Pg2 protecting group precursor A is

[0050] Preferably, the solvent A is a halogenated hydrocarbon solvent.

[0051] In a certain embodiment of the present invention, the Pg2 protecting group precursor B is R 5 Preferably is

[0052]

[0053] n is preferably 0 or 1;

[0054] R 2 Preferably is isopropyl;

[0055] Preferably, the Pg2 protecting group precursor B is

[0056] In one embodiment of the present invention, in the preparation method of Compound III, in the substitution reaction B, the molar ratio of the Pg2 protecting group precursor B to Compound II is preferably 1 - 5, such as 1.3, 3.5 or 4.

[0057] In one embodiment of the present invention, in the preparation method of Compound III, in the substitution reaction B, the molar ratio of the Pg2 protecting group precursor B to Compound II is 1 - 15, such as 1, 1.1, 1.3, 3.5, 4 or 8.

[0058] In one embodiment of the present invention, when the R 5 is , in the preparation method of Compound III, in the substitution reaction B, the molar ratio of the Pg2 protecting group precursor B to Compound II is 1 - 2, such as 1.3.

[0059] In one embodiment of the present invention, when the R 5 is , in the preparation method of Compound III, in the substitution reaction B, the molar ratio of the alkyl tertiary amine base to Compound II is 1 - 3, such as 2 or 2.6.

[0060] In one embodiment of the present invention, when the R 5 is , in the preparation method of Compound III, in the substitution reaction B, the concentration of Compound II in the halogenated hydrocarbon solvent is 0.20 - 0.30 mol / L, such as 0.23 mol / L or 0.24 mol / L.

[0061] In one embodiment of the present invention, when the R 5 is , in the preparation method of Compound III, in the substitution reaction B, the Base in Compound II and Compound III is

[0062] In the preparation method of Compound III, in the substitution reaction B, the alkyl tertiary amine base is preferably triethylamine or DIPEA.

[0063] In the preparation method of Compound III, in the substitution reaction B, the molar ratio of the alkyl tertiary amine base to Compound II is preferably 1.5 - 5, such as 2, 2.6, 4.4 or 4.5.

[0064] In one embodiment of the present invention, in the substitution reaction B, the molar ratio of the alkyl tertiary amine base to Compound II is preferably 1.5 - 15, such as 2, 2.6, 4.4, 4.5, 5, 9 or 10.

[0065] In the preparation method of the said Compound III, in the substitution reaction B, the halogenated hydrocarbon solvent is preferably DCM.

[0066] In the preparation method of the said Compound III, in the substitution reaction B, the concentration of the Compound II in the halogenated hydrocarbon solvent is preferably 0.1 - 0.30 mol / L, such as 0.11 mol / L, 0.13 mol / L, 0.17 mol / L, 0.18 mol / L, 0.23 mol / L or 0.24 mol / L.

[0067] In a certain embodiment of the present invention, in the substitution reaction B, the concentration of the Compound II in the halogenated hydrocarbon solvent is 0.1 - 1.5 mol / L, such as 0.11 mol / L, 0.13 mol / L, 0.15 mol / L, 0.17 mol / L, 0.18 mol / L, 0.23 mol / L, 0.24 mol / L, 0.26 mol / L, 0.57 mol / L or 1.27 mol / L.

[0068] In the preparation method of the said Compound III, the temperature of the substitution reaction B is preferably 10 - 30 °C, more preferably 20 - 30 °C, further preferably 20 - 25 °C, such as 20 °C or 25 °C.

[0069] In a certain embodiment of the present invention, the temperature of the substitution reaction B is 10 - 40 °C, such as 15 °C, 20 °C, 25 °C or 30 °C.

[0070] In the preparation method of the said Compound III, the progress of the substitution reaction B is monitored by conventional detection methods in the art (such as HPLC or LC - MS), and generally the disappearance or non - reaction of the Compound II in the reaction solution is taken as the end point of the reaction. The reaction time of the substitution reaction B is preferably 1 - 65 h, such as 1 h, 12 h, 20 h, 22 h or 64 h.

[0071] In a certain embodiment of the present invention, the reaction time of the substitution reaction B is 1 - 200 h, such as 1 h, 10 h, 12 h, 16 h, 20 h, 22 h, 64 h or 120 h.

[0072] In a certain embodiment of the present invention, the substitution reaction B includes one or more of the following post - treatment steps: liquid - liquid separation, washing the organic phase, concentration, column chromatography separation, liquid chromatography separation.

[0073] Among them, for the liquid - liquid separation, it is preferably to mix the reaction solution with an aqueous solution mixture of sodium carbonate and sodium bicarbonate and then perform liquid - liquid separation.

[0074] Among them, for washing the organic phase, it is preferably to wash with an aqueous solution mixture of sodium bicarbonate and sodium carbonate.

[0075] Among them, the column chromatography separation is preferably eluted with a mixed solvent of petroleum ether, ethyl acetate and triethylamine.

[0076] Among them, the liquid chromatography separation is preferably eluted with acetonitrile as the mobile phase.

[0077] In a certain embodiment of the present invention, the substitution reaction B includes one or more of the following post-treatment steps: liquid separation, washing the organic phase, concentration, column chromatography separation, and liquid chromatography separation.

[0078] Among them, the liquid separation is preferably carried out by mixing the reaction solution with an aqueous sodium bicarbonate solution and then separating the liquid, or by mixing the reaction solution with a mixed aqueous solution of sodium carbonate and sodium bicarbonate and then separating the liquid;

[0079] Among them, the washing of the organic phase is preferably carried out with an aqueous sodium bicarbonate solution, or with a mixed aqueous solution of sodium bicarbonate and sodium carbonate;

[0080] Among them, the column chromatography separation is preferably eluted with a mixed solvent of petroleum ether, ethyl acetate and triethylamine, or with a mixed solvent of dichloromethane, methanol and triethylamine.

[0081] Among them, the liquid chromatography separation is preferably eluted with acetonitrile as the mobile phase.

[0082] In a certain embodiment of the present invention, in the preparation method of the compound III, in the substitution reaction B, the compound II is

[0083]

[0084] Correspondingly, the compound III is

[0085]

[0086] In a certain embodiment of the present invention, the substitution reaction B is any of the following cases:

[0087] (1) The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA;

[0088] (2) The Pg2 protecting group precursor B is The alkyl tertiary amine base is triethylamine;

[0089] (3) The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; Preferably, the substitution reaction B is any of the following cases:

[0090] (1) The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 4, and the molar ratio of the alkyl tertiary amine base to Compound II is 4.4;

[0091] (2) The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 3.5, and the molar ratio of the alkyl tertiary amine base to Compound II is 4.4;

[0092] (3) The Pg2 protecting group precursor B is The alkyl tertiary amine base is triethylamine; the molar ratio of the Pg2 protecting group precursor B to Compound II is 1.3, and the molar ratio of the alkyl tertiary amine base to Compound II is 2;

[0093] (4) The Pg2 protecting group precursor B is The alkyl tertiary amine base is triethylamine; the molar ratio of the Pg2 protecting group precursor B to Compound II is 1.3, and the molar ratio of the alkyl tertiary amine base to Compound II is 2.6;

[0094] (5) The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 8, and the molar ratio of the alkyl tertiary amine base to Compound II is 9;

[0095] (6) The Pg2 protecting group precursor B is The alkyl tertiary amine base is triethylamine; the molar ratio of the Pg2 protecting group precursor B to Compound II is 1, and the molar ratio of the alkyl tertiary amine base to Compound II is 2;

[0096] (7) The Pg2 protecting group precursor B is The alkyl tertiary amine base is triethylamine; the molar ratio of the Pg2 protecting group precursor B to Compound II is 1.1, and the molar ratio of the alkyl tertiary amine base to Compound II is 2;

[0097] (8) The Pg2 protecting group precursor B is The alkyl tertiary amine base is triethylamine; the molar ratio of the Pg2 protecting group precursor B to Compound II is 1.3, and the molar ratio of the alkyl tertiary amine base to Compound II is 2;

[0098] (9) The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 8, and the molar ratio of the alkyl tertiary amine base to Compound II is 10;

[0099] (10) The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 3.5, and the molar ratio of the alkyl tertiary amine base to Compound II is 4.5;

[0100] (11) The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 4, and the molar ratio of the alkyl tertiary amine base to Compound II is 5;

[0101] More preferably, the substitution reaction B is any of the following cases:

[0102] (1) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 4, and the molar ratio of the alkyl tertiary amine base to Compound II is 4.4;

[0103] (2) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 3.5, and the molar ratio of the alkyl tertiary amine base to Compound II is 4.4;

[0104] (3) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 4, and the molar ratio of the alkyl tertiary amine base to Compound II is 4.4;

[0105] (4) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is triethylamine; the molar ratio of the Pg2 protecting group precursor B to Compound II is 1.3, and the molar ratio of the alkyl tertiary amine base to Compound II is 2;

[0106] (5) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is triethylamine; the molar ratio of the Pg2 protecting group precursor B to Compound II is 1.3, and the molar ratio of the alkyl tertiary amine base to Compound II is 2.6;

[0107] (6) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 8, and the molar ratio of the alkyl tertiary amine base to Compound II is 9;

[0108] (7) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is triethylamine; the molar ratio of the Pg2 protecting group precursor B to Compound II is 1, and the molar ratio of the alkyl tertiary amine base to Compound II is 2;

[0109] (8) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is triethylamine; the molar ratio of the Pg2 protecting group precursor B to Compound II is 1.1, and the molar ratio of the alkyl tertiary amine base to Compound II is 2;

[0110] (9) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is triethylamine; the molar ratio of the Pg2 protecting group precursor B to Compound II is 1.3, and the molar ratio of the alkyl tertiary amine base to Compound II is 2;

[0111] (10) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 8, and the molar ratio of the alkyl tertiary amine base to Compound II is 10;

[0112] (11) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 3.5, and the molar ratio of the alkyl tertiary amine base to Compound II is 4.5;

[0113] (12) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 3.5, and the molar ratio of the alkyl tertiary amine base to Compound II is 4.5;

[0114] (13) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 3.5, and the molar ratio of the alkyl tertiary amine base to Compound II is 4.5;

[0115] (14) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 4, and the molar ratio of the alkyl tertiary amine base to Compound II is 5;

[0116] (15) Compound II is The Pg2 protecting group precursor B is The alkyl tertiary amine base is DIPEA; the molar ratio of the Pg2 protecting group precursor B to Compound II is 3.5, and the molar ratio of the alkyl tertiary amine base to Compound II is 4.5.

[0117] In the preparation method of Compound III, in the substitution reaction A, the R 1 is preferably H, acetyl or isobutyryl.

[0118] In the preparation method of Compound III, in the substitution reaction A, the molar ratio of DMF-DMA to Compound II is preferably 1 - 5, more preferably 1.5 - 4.5, such as 3 or 4.

[0119] In a certain embodiment of the present invention, in the substitution reaction A, the molar ratio of DMF-DMA to Compound II is 1 - 5, such as 2.5, 3, 4 or 4.5.

[0120] In a certain embodiment of the present invention, in the substitution reaction A, the molar ratio of the Pg2 protecting group precursor A to Compound II is 1 - 10, preferably 1 - 6, such as 2.5, 3, 4, 4.5 or 6.

[0121] In the preparation method of Compound III, in the substitution reaction A, the alcohol solvent is preferably a C1-C4 alcohol solvent, such as methanol.

[0122] In a certain embodiment of the present invention, in the substitution reaction A, in the solvent A, the halogenated hydrocarbon solvent is DCM.

[0123] In one embodiment of the present invention, in the substitution reaction A, in the solvent A, the nitrogen atom in the nitrogen-containing aromatic compound solvent is located on the aromatic ring of the aromatic compound.

[0124] In one embodiment of the present invention, in the substitution reaction A, in the solvent A, the nitrogen-containing aromatic compound solvent is pyridine.

[0125] In the preparation method of the compound III, in the substitution reaction A, the concentration of the compound II in the alcohol solvent is preferably 0.05 - 0.25 mol / L, more preferably 0.1 - 0.2 mol / L, such as 0.11 mol / L, 0.12 mol / L, 0.16 mol / L or 0.18 mol / L.

[0126] In one embodiment of the present invention, in the substitution reaction A, the concentration of the compound II in the solvent A is 0.05 - 0.25 mol / L, preferably 0.1 - 0.2 mol / L, such as 0.10 mol / L, 0.11 mol / L, 0.12 mol / L, 0.13 mol / L, 0.14 mol / L, 0.16 mol / L or 0.18 mol / L.

[0127] In the preparation method of the compound III, the temperature of the substitution reaction A is preferably 10 - 30 °C, more preferably 20 - 30 °C, further preferably 20 - 25 °C, such as 20 °C or 25 °C.

[0128] In one embodiment of the present invention, in the preparation method of the compound III, the temperature of the substitution reaction A is 10 - 40 °C, such as 15 °C, 20 °C, 25 °C, 30 °C or 35 °C.

[0129] In the preparation method of the compound III, the progress of the substitution reaction A is monitored by conventional detection methods in the art (such as HPLC or LC-MS), and generally the disappearance or no longer reaction of the compound II in the reaction solution is used as the reaction end point. The reaction time of the substitution reaction A is preferably 3 - 20 h, such as 3 h or 16 h.

[0130] In one embodiment of the present invention, the reaction time of the substitution reaction A is 3 - 30 h, such as 3 h, 12 h, 16 h or 24 h.

[0131] In one embodiment of the present invention, the substitution reaction A includes one or more of the following post-treatment steps: concentration and column chromatography separation.

[0132] Among them, the column chromatography separation is preferably eluted with one or more of dichloromethane, methanol, petroleum ether, ethyl acetate and triethylamine. For example, it is eluted with a mixed solvent of dichloromethane, methanol and triethylamine, or eluted with a mixed solvent of petroleum ether, ethyl acetate and triethylamine.

[0133] In a certain embodiment of the present invention, the substitution reaction A includes one or more of the following post-treatment steps:

[0134] Dilution, washing, concentration and column chromatography separation;

[0135] The dilution is preferably carried out by diluting the reaction solution with ethyl acetate or methyl tert-butyl ether;

[0136] The washing is preferably carried out by washing the diluted solution with semi-saturated brine, and taking the organic phase;

[0137] The column chromatography separation is preferably eluted with one or more of dichloromethane, methanol, petroleum ether, ethyl acetate, acetonitrile and triethylamine. For example, it is eluted with a mixed solvent of dichloromethane, methanol and triethylamine, or eluted with a mixed solvent of petroleum ether, ethyl acetate and triethylamine, or eluted with acetonitrile.

[0138] In a certain embodiment of the present invention, when the R 1 is H,

[0139] in the substitution reaction A, the compound II is

[0140] Correspondingly, the compound III is

[0141] In a certain embodiment of the present invention, when the R 1 is H,

[0142] in the substitution reaction A, the compound II is Correspondingly, the compound III is

[0143] In a certain embodiment of the present invention, when the R 1 is acetyl or isobutyryl,

[0144] in the substitution reaction A, the compound II is

[0145]

[0146] Correspondingly, the compound III is

[0147] In a certain embodiment of the present invention, the substitution reaction A is any of the following cases:

[0148] (1) Compound II is The molar ratio of the DMF-DMA to Compound II is 4;

[0149] (2) Compound II is The molar ratio of the DMF-DMA to Compound II is 4;

[0150] (3) Compound II is The molar ratio of the DMF-DMA to Compound II is 3;

[0151] (4) Compound II is The molar ratio of the DMF-DMA to Compound II is 1.5;

[0152] (5) Compound II is The molar ratio of the DMF-DMA to Compound II is 2.5;

[0153] (6) Compound II is The molar ratio of the DMF-DMA to Compound II is 3;

[0154] (7) Compound II is The molar ratio of the DMF-DMA to Compound II is 4;

[0155] (8) Compound II is The molar ratio of the DMF-DMA to Compound II is 3;

[0156] (9) Compound II is The molar ratio of the DMF-DMA to Compound II is 4;

[0157] (10) Compound II is The molar ratio of the DMF-DMA to Compound II is 3;

[0158] (11) Compound II is The molar ratio of the DMF-DMA to Compound II is 3;

[0159] (12) Compound II is The molar ratio of the DMF-DMA to Compound II is 3;

[0160] (13) The compound II is The molar ratio of the DMF-DMA to the compound II is 2.5;

[0161] (14) The compound II is The molar ratio of the DMF-DMA to the compound II is 4;

[0162] (15) The compound II is The molar ratio of the DMF-DMA to the compound II is 4.5;

[0163] (16) The compound II is The molar ratio of the N,N-diethylformamide dimethyl acetal to the compound II is 6;

[0164] (17) The compound II is The molar ratio of the N,N-dimethylacetamide dimethyl acetal to the compound II is 6;

[0165] (18) The compound II is The molar ratio of the N,N-dibutylformamide dimethyl acetal to the compound II is 6;

[0166] (19) The compound II is The molar ratio of the 2,2-dimethoxy-1-methylpyrrolidine to the compound II is 6.

[0167] In a certain embodiment of the present invention, the method for preparing the compound of formula III further comprises the following steps:

[0168] In the presence of an organic base, in a solvent, the compound I is subjected to a deprotection reaction to obtain the compound II;

[0169]

[0170] wherein, the organic base is ammonia and / or alkylamine;

[0171] The solvent is an ether solvent and / or an alcohol solvent;

[0172] R 1 is H;

[0173] Base is (* The position is directly connected to R 1 or Pg1);

[0174] Pg1 is an alkyl-C(=O)- of C1-C4 or an aryl-C(=O)- of C6-C 14 ;

[0175] Pg1 is directly connected to the amino group of Base;

[0176] R is H, C1-C4 alkoxy, -OTBS, -OTOM, halogen or -O-MOE.

[0177] In the deprotection reaction, the C1-C4 alkyl-C(=O)- is preferably acetyl or isobutyryl.

[0178] In the deprotection reaction, the C6-C 14 aryl-C(=O)- is preferably benzoyl.

[0179] In the deprotection reaction, the organic base is preferably ammonia or alkylamine; the alkylamine is preferably methylamine, ethylamine or triethylamine.

[0180] In the deprotection reaction, the molar ratio of the organic base to Compound I is preferably 5-40, such as 10, 12, 17.2, 17.4, 17.6, 19.8, 34 or 36.

[0181] In one embodiment of the present invention, in the deprotection reaction, the molar ratio of the organic base to Compound I is 5-80, such as 10, 12, 17.2, 17.4, 17.6, 19.8, 20, 34, 36, 58 or 59.

[0182] In the deprotection reaction, the solvent is preferably an ether solvent or an alcohol solvent; the ether solvent is preferably THF; the alcohol solvent is preferably a C1-C4 alcohol solvent, such as methanol.

[0183] In the deprotection reaction, the concentration of Compound I in the solvent is preferably 0.05-0.35 mol / L, such as 0.06 mol / L, 0.1 mol / L, 0.11 mol / L, 0.12 mol / L, 0.13 mol / L, 0.25 mol / L or 0.27 mol / L.

[0184] The temperature of the deprotection reaction is preferably 15-30 °C, more preferably 20-25 °C, such as 20 °C or 25 °C.

[0185] In one embodiment of the present invention, the temperature of the deprotection reaction is 10-35 °C, such as 15 °C, 20 °C, 25 °C or 30 °C.

[0186] The progress of the deprotection reaction is monitored by conventional detection methods in the art (such as HPLC or LC-MS), and generally, the disappearance or no longer reaction of the compound I in the reaction solution is taken as the end point of the reaction. The reaction time of the deprotection reaction is preferably 3 - 65 h, such as 3 h, 12 h, 16 h, 40 h or 60 h.

[0187] In a certain embodiment of the present invention, the deprotection reaction includes one or more of the following post-treatment steps:

[0188] Concentration, liquid separation, recrystallization, column chromatography separation.

[0189] Among them, for the liquid separation, it is preferred to mix the reaction solution with water and methyl tert-butyl ether and then perform liquid separation to take the organic phase.

[0190] Among them, for the recrystallization, it is preferred to perform recrystallization with methyl tert-butyl ether and n-hexane, or with ethyl acetate and n-heptane.

[0191] Among them, for the column chromatography separation, it is preferred to elute with a mixed solvent of petroleum ether, ethyl acetate and / or triethylamine.

[0192] In a certain embodiment of the present invention, in the deprotection reaction, in the post-treatment step, the liquid separation is carried out through the following steps: Mix the reaction solution with methyl tert-butyl ether, wash with semi-saturated brine (such as washing 3 times), and perform liquid separation to take the organic phase.

[0193] In a certain embodiment of the present invention, when Pg1 is benzoyl, it satisfies one or more of the following conditions:

[0194] (1) In the deprotection reaction, the organic base is an alkylamine; the alkylamine is methylamine or ethylamine;

[0195] (2) In the deprotection reaction, the molar ratio of the organic base to the compound I is 15 - 40, such as 17.2, 17.6, 19.8 or 36;

[0196] (3) In the deprotection reaction, the solvent is an ether solvent; the ether solvent is preferably THF;

[0197] (4) In the deprotection reaction, the concentration of the compound I in the solvent is 0.05 - 0.15 mol / L, such as 0.06 mol / L, 0.10 mol / L, 0.11 mol / L or 0.12 mol / L;

[0198] (5) In the deprotection reaction, the temperature is 20 - 25 °C, such as 20 °C or 25 °C;

[0199] (6) The reaction time of the deprotection reaction is preferably 3 - 15 h, such as 3 h or 12 h.

[0200] In a certain embodiment of the present invention, when Pg1 is an acetyl group, it satisfies one or more of the following conditions:

[0201] (1) In the deprotection reaction, the organic base is an alkylamine, such as triethylamine;

[0202] (2) In the deprotection reaction, the molar ratio of the organic base to Compound I is 5 - 15, such as 10;

[0203] (3) In the deprotection reaction, the solvent is an alcohol solvent; the alcohol solvent is preferably a C1 - C4 alcohol solvent, such as methanol;

[0204] (4) In the deprotection reaction, the concentration of Compound I in the solvent is 0.20 - 0.30 mol / L, such as 0.25 mol / L or 0.27 mol / L;

[0205] (5) The temperature of the deprotection reaction is 20 - 25 °C, such as 25 °C;

[0206] (6) The reaction time of the deprotection reaction is preferably 5 - 15 h, such as 12 h.

[0207] In a certain embodiment of the present invention, when Pg1 is an isobutyryl group, it satisfies one or more of the following conditions:

[0208] (1) In the deprotection reaction, the molar ratio of the organic base to Compound I is 10 - 40, such as 12, 17.4 or 34;

[0209] (2) In the deprotection reaction, the solvent is an alcohol solvent or an ether solvent; the alcohol solvent is preferably a C1 - C4 alcohol solvent, such as methanol; the ether solvent is preferably THF;

[0210] (3) In the deprotection reaction, the concentration of Compound I in the solvent is 0.08 - 0.15 mol / L, such as 0.10 mol / L, 0.11 mol / L or 0.13 mol / L;

[0211] (4) The temperature of the deprotection reaction is 20 - 25 °C, such as 25 °C;

[0212] (5) The reaction time of the deprotection reaction is preferably 12 - 60 h, such as 16 h, 40 h or 60 h.

[0213] In a certain embodiment of the present invention, in the deprotection reaction, the compound I is

[0214]

[0215] Correspondingly, the compound II is

[0216]

[0217] In a certain embodiment of the present invention, the deprotection reaction is any of the following cases:

[0218] (1) The organic base is methylamine and the solvent is THF;

[0219] (2) The organic base is ethylamine and the solvent is THF;

[0220] (3) The organic base is triethylamine and the solvent is methanol;

[0221] (4) The organic base is ammonia and the solvent is methanol;

[0222] Preferably, the deprotection reaction is any of the following cases:

[0223] (1) The compound I is The organic base is methylamine and the solvent is THF;

[0224] (2) The compound I is The organic base is ethylamine and the solvent is THF;

[0225] (3) The compound I is The organic base is methylamine and the solvent is THF;

[0226] (4) The compound I is The organic base is ethylamine and the solvent is THF;

[0227] (5) The compound I is The organic base is triethylamine and the solvent is methanol;

[0228] (6) The compound I is The organic base is triethylamine and the solvent is methanol;

[0229] (7) The compound I is The organic base is ammonia and the solvent is methanol;

[0230] (8) The compound I is The organic base is ammonia and the solvent is methanol;

[0231] (9) The compound I is The organic base is ethylamine and the solvent is THF;

[0232] (10) The compound I is The organic base is triethylamine and the solvent is methanol;

[0233] (11) The compound I is The organic base is ammonia and the solvent is methanol;

[0234] (12) The compound I is The organic base is ammonia and the solvent is methanol;

[0235] (13) The compound I is The organic base is triethylamine and the solvent is methanol;

[0236] (14) The compound I is The organic base is ethylamine and the solvent is THF;

[0237] (15) The compound I is The organic base is ethylamine and the solvent is THF.

[0238] The present invention also provides a method for preparing a compound of formula II, which comprises the following steps:

[0239] In the presence of ammonia and / or alkylamine, in a solvent, the compound I as shown is subjected to a deprotection reaction to obtain the compound II as shown;

[0240]

[0241] Among them, the conditions and operations are preferably the same as those described in the above deprotection reaction.

[0242] The present invention also provides a compound of formula IV-1 and formula IV-2:

[0243]

[0244] The present invention also provides an application of the compound of formula IV-1 and formula IV-2 as an amino protecting reagent; in the said application, the compound of formula IV-1 and formula IV-2 replaces the hydrogen atom in -NH- or -NH2 in the nitrogen-containing organic compound with an amino protecting group;

[0245] The amino protecting group is

[0246] The nitrogen-containing organic compound contains a -NH- or -NH2 structure.

[0247] In one embodiment of the present invention, the application includes the following steps:

[0248] In the presence of an alkyl tertiary amine base and in a halogenated hydrocarbon solvent, the nitrogen-containing organic compound and the compound of formula IV-1 or formula IV-2 are subjected to substitution reaction B to obtain a product in which the hydrogen atom in -NH- or -NH2 in the nitrogen-containing organic compound is substituted with the amino protecting group;

[0249] Among them, the conditions and operations are preferably the same as those in the above substitution reaction B;

[0250] The nitrogen-containing organic compound is preferably an amino nucleoside;

[0251] The amino nucleoside is preferably

[0252] wherein Base is (*The position is directly connected to R 1 connected);

[0253] R 1 is H;

[0254] R is H, C1-C4 alkoxy, -OTBS, -OTOM, halogen or -O-MOE;

[0255] The amino nucleoside is more preferably

[0256]

[0257] The present invention also provides a method for preparing the compound of formula IV-1, which includes the following steps:

[0258] In the presence of a condensing agent and in a halogenated hydrocarbon solvent, p-nitrophenol and 4-isopropylphenoxyacetic acid are subjected to a condensation reaction to obtain the compound IV-1.

[0259] In the condensation reaction, the condensing agent is preferably N,N-diisopropylcarbodiimide.

[0260] In the condensation reaction, the molar ratio of the condensing agent to the 4-isopropylphenoxyacetic acid is preferably 1.

[0261] In the condensation reaction, the halogenated hydrocarbon solvent is preferably dichloromethane.

[0262] In the condensation reaction, the concentration of 4-isopropylphenoxyacetic acid in the halogenated hydrocarbon solvent is preferably 0.51 mol / L.

[0263] In the condensation reaction, the molar ratio of p-nitrophenol to 4-isopropylphenoxyacetic acid is preferably 1.

[0264] The temperature of the condensation reaction is preferably 20 °C.

[0265] The time of the condensation reaction is preferably 12 h.

[0266] The condensation reaction may include the following post-treatment steps: filtration, concentration, and column chromatography separation.

[0267] The present invention also provides a method for preparing the compound IV-2, which includes the following steps:

[0268] In the presence of a condensing agent, in a halogenated hydrocarbon solvent, pentafluorophenol and 4-isopropylphenoxyacetic acid are subjected to a condensation reaction to obtain the compound IV-2.

[0269] In the condensation reaction, the condensing agent is preferably N,N-diisopropylcarbodiimide.

[0270] In the condensation reaction, the molar ratio of the condensing agent to 4-isopropylphenoxyacetic acid is preferably 1.

[0271] In the condensation reaction, the halogenated hydrocarbon solvent is preferably dichloromethane.

[0272] In the condensation reaction, the concentration of 4-isopropylphenoxyacetic acid in the halogenated hydrocarbon solvent is preferably 0.51 mol / L.

[0273] In the condensation reaction, the molar ratio of pentafluorophenol to 4-isopropylphenoxyacetic acid is preferably 1.

[0274] The temperature of the condensation reaction is preferably 20 °C.

[0275] The time of the condensation reaction is preferably 16 h.

[0276] The condensation reaction may include the following post-treatment steps: filtration, concentration, and column chromatography separation.

[0277] In a certain embodiment of the present invention, in any of the substitution reactions A, any of the substitution reactions B, any of the deprotection reactions, and any of the condensation reactions, the materials used are limited to those mentioned in this technical solution and do not involve materials not mentioned.

[0278] Unless otherwise specified, the various terms in the present invention may have the following definitions:

[0279] Those skilled in the art can understand that, according to the conventions used in the art, in the structural formulas of the groups described in the present invention, the means that the corresponding group is connected to other fragments or groups in the compound through this site.

[0280] In this article, the substituents used can be added with a single dash "-", indicating that the named substituent is connected to the parent part through a single bond.

[0281] The term "alkyl" refers to a straight-chain or branched-chain alkyl group having a specified number of carbon atoms (for example, C1-C6). Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, etc.

[0282] The term "alkoxy" refers to the group R X -O-, where the definition of R X is the same as the term "alkyl".

[0283] The term "carbocyclic heterocycle" refers to a saturated cyclic group having a specified number of ring atoms (for example, 3-6 membered), a specified number of heteroatoms (for example, 1 or 2), and a specified type of heteroatoms (1, 2, or 3 of N, O, and S). Examples of carbocyclic heterocycles include, but are not limited to, oxiranyl, aziridinyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, morpholinyl, or piperazinyl, etc.

[0284] "Molar ratio" refers to the numerical value of the amount of substance of one substance to the amount of substance of another substance. For example, the meaning of "the molar ratio of the Pg2 protecting group precursor B to compound II is 1-15" is "the ratio of the amount of substance of the Pg2 protecting group precursor B to the amount of substance of the compound II is (1:1)-(15:1)".

[0285] On the basis of not violating the common knowledge in the art, the above preferred conditions can be combined arbitrarily to obtain various preferred examples of the present invention.

[0286] The reagents and raw materials used in the present invention are all commercially available.

[0287] The positive and progressive effects of the present invention are as follows: A new strategy for synthesizing phosphoramidite monomers with easily removable amino protecting groups is provided, which has one or more of the following advantages: fewer reaction steps; low cost; high overall yield; mild conditions and high safety; easy post-treatment; suitable for industrial production; and can rapidly and massively prepare phosphoramidite monomers with easily removable base protecting groups. Detailed Embodiments

[0288] The present invention will be further described below by way of examples, but the present invention is not limited to the scope of the described examples. For the experimental methods without specific conditions in the following examples, they are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0289] Example 1 Synthesis of Intermediate A, Intermediate IV-1, Intermediate IV-2, Intermediate IV-3 and Compound WX001

[0290] Synthesis route of Intermediate A:

[0291]

[0292] Compound A-1 (25 g, 164.31 mmol, 1 eq) was added to dichloromethane (250 mL), followed by addition of pentafluorophenol (30.24 g, 164.31 mmol, 1 eq) and N,N-diisopropylcarbodiimide (20.74 g, 164.31 mmol, 25.44 mL, 1 eq). The reaction was stirred at 20 °C for 12 hours. The reaction was detected to be complete by LCMS, water (300 mL) was added, and liquid separation extraction was carried out. The organic phase was dried with anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 100:1 to 0:1) to obtain white solid Compound A (48 g, yield: 92%).

[0293] 1 1H NMR (400 MHz, ACETONITRILE-d3) δ = 7.37–7.33 (m, 2H), 7.06–6.99 (m, 3H), 5.12 (s, 2H).

[0294] Synthesis route of Intermediate IV-1:

[0295]

[0296] Compound IV-1-1 (100 g, 514.86 mmol, 1 eq) was added to dichloromethane (1000 mL), followed by addition of p-nitrophenol (71.62 g, 514.86 mmol, 1 eq) and N,N-diisopropylcarbodiimide (64.98 g, 514.86 mmol, 1 eq). The reaction was stirred at 20 °C for 12 hours. The reaction was detected to be complete by TLC, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 100:1 to 0:1) to obtain yellow solid Compound IV-1 (130 g, yield: 80%).

[0297] 11H NMR (400 MHz, CHLOROFORM-d1) δ = 8.30–8.26 (m, 2H), 7.35–7.32 (m, 2H), 7.21–7.19 (m, 2H), 6.94–6.92 (m, 2H), 4.90 (s, 2H), 2.93–2.86 (m, 1H), 1.25 (s, 3H), 1.24 (s, 3H).

[0298] Synthetic route of intermediate IV-2:

[0299]

[0300] Compound IV-1-1 (10 g, 51.49 mmol, 1 eq) was added to dichloromethane (100 mL), followed by addition of pentafluorophenol (9.48 g, 51.49 mmol, 1 eq) and N,N-diisopropylcarbodiimide (6.50 g, 51.49 mmol, 1 eq). The reaction was stirred at 20 °C for 16 h. The reaction was monitored by TLC until completion, and the reaction mixture was filtered. The filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 100:1 to 0:1) to give compound IV-2 as a colorless oil (12 g, yield: 64.6%).

[0301] 1 1H NMR (400 MHz, CHLOROFORM-d1) δ = 7.21-7.19 (m, 2H), 6.92–6.89 (m, 2H), 4.98 (s, 2H), 2.93–2.86 (m, 1H), 1.25 (s, 3H), 1.24 (s, 3H).

[0302] Synthetic route of intermediate IV-3:

[0303]

[0304] Compound IV-1-1 (10 g, 51.49 mmol, 1 eq) was added to dichloromethane (100 mL), followed by addition of N-hydroxyphthalimide (8.40 g, 51.49 mmol, 1 eq) and N,N-diisopropylcarbodiimide (6.50 g, 51.49 mmol, 1 eq). The reaction was stirred at 20 °C for 16 h. The reaction was monitored by TLC until completion, and the reaction mixture was filtered. The filtrate was concentrated under reduced pressure to give the crude product of compound IV-3 as a yellow solid (12 g).

[0305] Synthesis of compound WX001

[0306]

[0307] Step 1:

[0308] Compound 1-1 (20 g, 20.24 mmol, 1 eq) was added to a flask containing methylamine (2 M solution in tetrahydrofuran, 200 mL, 19.76 eq). The reaction was stirred at 20 °C for 3 hours. The reaction was monitored by LCMS and found to be complete. The reaction mixture was concentrated under reduced pressure to obtain crude compound 1-2 (23 g) as a colorless oil.

[0309] Step 2:

[0310] Compound 1-2 (20 g, 22.62 mmol, 1 eq) and N,N-dimethylformamide dimethyl acetal (10.78 g, 90.49 mmol, 4 eq) were added to a flask containing methanol (140 mL). The reaction was stirred at 20 °C for 3 hours. The reaction was monitored by LCMS and found to be complete. The reaction mixture was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 3:1 to 0:1, with 0.5% triethylamine) to obtain off-white solid compound WX001 (14 g, yield: 65%, purity: 99%).

[0311] 1 H NMR (400 MHz, ACETONITRILE-d3) δ = 8.89 (d, J = 3.6 Hz, 1H), 8.29 (d, J = 14.8 Hz, 1H), 8.08 (d, J = 11.2 Hz, 1H), 7.47 - 7.27 (m, 9H), 6.85–6.81 (m, 4H), 5.97 (dd, J = 4.4 Hz, J = 2.4 Hz, 1H), 5.14–5.07 (m, 1H), 4.48–4.46 (m, 2H), 3.76 - 3.75 (m, 1H), 3.65 (s, 6H), 3.64 - 3.31 (m, 5H), 3.16 - 3.15 (m, 6H), 2.67 - 2.44 (m, 2H), 1.19 - 1.09 (m, 12H), 0.74 (s, 9H), -0.03--0.2 (dd, J = 11.6 Hz, J = 9.6 Hz, 6H)

[0312] LCMS (ESI) m / z: 939.2 [M+H] +

[0313] Example 2 Synthesis of Compound WX002

[0314]

[0315] Step 1:

[0316] Compound 2-1 (45 g, 51.37 mmol, 1 eq) was added to a flask containing ethylamine (2 M solution in tetrahydrofuran, 450 mL), and the reaction was stirred at 25 °C for 12 h. The reaction was monitored by LCMS and was found to be complete. The reaction mixture was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 50:1 to 0:1, with 0.5% triethylamine) to obtain a white solid compound 2-2 (34.8 g, yield: 86%, purity: 98%).

[0317] Step 2:

[0318] Compound 2-2 (34.8 g, 45.09 mmol, 1 eq) and N,N-dimethylformamide dimethyl acetal (10.75 g, 90.18 mmol, 4 eq) were added to a flask containing methanol (245 mL), and the reaction was stirred at 25 °C for 3 h. The reaction was monitored by LCMS and was found to be complete. The reaction mixture was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 50:1 to 0:1, with 0.5% triethylamine) to obtain a pale yellow solid compound WX002 (34 g, yield: 89%, purity: 98%).

[0319] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 8.89 (s, 1H), 8.42 (d, J = 4.0 Hz, 1H), 8.16 (s, 1H), 7.32 - 7.18 (m, 9H), 6.77–6.74 (m, 4H), 6.32–6.27 (m, 1H), 6.24–5.81 (m, 1H), 5.40–5.20 (m, 1H), 4.25–4.24 (m, 1H), 3.89–3.71 (m, 1H), 3.65 (s, 6H), 3.63 - 3.23 (m, 5H), 3.13 (s, 6H), 2.66 - 2.49 (m, 2H), 1.20 - 1.06 (m, 12H)

[0320] LCMS (ESI) m / z: 827.8 [M+H] +

[0321] Example 3 Synthesis of Compound WX003

[0322] Synthesis route:

[0323]

[0324] Step 1:

[0325] Compound 3-1 (100 g, 112.62 mmol, 1 eq) was added to a flask containing methylamine (2 M solution in tetrahydrofuran, 2.03 L, 36 eq). The reaction was stirred at 20 °C for 3 hours. The reaction was monitored by LCMS and when it was complete, water (2 L) was added, followed by methyl tert-butyl ether (2 L). The layers were separated, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 100:1 to 0:1) to obtain a white solid compound 3-2 (83 g, yield: 92%, purity: 98%).

[0326] Step 2: WX003 was synthesized in a similar manner to Example 2.

[0327] Compound 3-2 (41 g, 52.30 mmol, 1 eq) and N,N-dimethylformamide dimethyl acetal (15.58 g, 130.76 mmol, 2.5 eq) were added to a flask containing methanol (410 mL). The reaction was stirred at 25 °C for 3 hours. The reaction was monitored by LCMS and when it was complete, the reaction mixture was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 50:1 to 0:1, with 0.3% triethylamine) to obtain a pale yellow solid compound WX003 (29 g, yield: 59%, purity: 98.2%).

[0328] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 8.88 (s, 1H), 8.38 - 8.36 (d, J = 7.6 Hz, 1H), 8.15 - 8.12 (d, J = 10 Hz, 1H), 7.43 - 7.16 (m, 9H), 6.82 - 6.77 (m, 4H), 6.10 - 6.09 (m, 1H), 4.85 - 4.67 (m, 2H), 4.35 - 4.30 (m, 1H), 3.79 - 3.70 (m, 1H), 3.72 (s, 6H), 3.71 - 3.70 (m, 3H), 3.48 - 3.41 (m, 4H), 3.33 - 3.27 (m, 1H), 3.13 (s, 6H), 2.70 - 2.52 (m, 2H), 1.20 - 1.07 (m, 12H)

[0329] LCMS (ESI) m / z: 899.4 [M+HCO3] -

[0330] Synthesis of Compound WX004 in Example 4

[0331] Synthetic route:

[0332]

[0333] Step 1:

[0334] Compound 4-1 (22 g, 25.64 mmol, 1 eq) was added to a flask containing ethylamine (2 M solution in tetrahydrofuran, 220 mL), and the reaction was stirred at 25 °C for 12 hours. The reaction was monitored by LCMS and found to be complete. The reaction mixture was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 3:1 to 0:1) to obtain a white solid compound 4-2 (12.8 g, yield: 67%, purity: 99%).

[0335] Step 2: WX004 was synthesized in a similar manner to Example 2.

[0336] Compound 4-2 (5 g, 6.63 mmol, 1 eq) and N,N-dimethylformamide dimethyl acetal (2.37 g, 19.89 mmol, 3 eq) were added to a flask containing methanol (50 mL), and the reaction was stirred at 15 °C for 12 hours. The reaction was monitored by LCMS and found to be complete. Ethyl acetate (50 mL) was added, and the mixture was washed twice with half-saturated brine (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 50:1 to 0:1, with 0.3% triethylamine) to obtain a white solid compound WX004 (4 g, yield: 74.5%, purity: 98.3%).

[0337] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 8.88 (s, 1H), 8.35 (s, 1H), 8.10 (s, 1H), 7.37 - 7.19 (m, 9H), 6.78 - 6.74 (m, 4H), 6.43 - 6.39 (m, 1H), 4.93 - 4.87 (m, 1H), 4.21 - 4.20 (m, 1H), 3.72 - 3.60 (m, 10H), 3.27 - 3.26 (m, 2H), 3.13 (s, 6H), 2.66 - 2.53 (m, 3H), 1.18 - 1.07 (m, 12H)

[0338] LCMS (ESI) m / z: 809.6 [M+H] +

[0339] Synthesis of Compound WX005 in Example 5

[0340] Synthesis route:

[0341]

[0342] Step 1:

[0343] Compound 5-1 (19 g, 21.06 mmol, 1 eq) and N,N-dimethylformamide dimethyl acetal (7.53 g, 63.19 mmol, 3 eq) were added to a flask containing methanol (190 mL). The reaction was stirred at 25 °C for 16 h. The reaction was monitored by LCMS and found to be complete. The reaction mixture was concentrated under reduced pressure. Methyl tert-butyl ether (100 mL) was added, and the mixture was washed twice with semi-saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 100:1 to 0:1, with 0.5% triethylamine) to obtain a white solid compound Compound WX005 (11 g, yield: 57%, purity: 98%).

[0344] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 8.67 (s, 1H), 7.98 - 7.88 (m, 1H), 7.49 - 7.24 (m, 10H), 6.89–6.85 (m, 4H), 6.01–5.91 (m, 1H), 5.69–5.66 (m, 1H), 4.36–4.25 (m, 2H), 3.76 (s, 6H), 3.60 - 3.52 (m, 6H), 3.12 - 3.05 (m, 6H), 2.65 - 2.44 (m, 2H), 1.15 - 1.00 (m, 12H), 0.90 (s, 9H), 0.13 - 0.11 (m, 6H)

[0345] LCMS (ESI) m / z: 915.3 [M+H] +

[0346] Synthesis of Compound WX006 in Example 6

[0347] Synthesis route:

[0348]

[0349] WX006 was synthesized by a method similar to that in Example 5

[0350] Compound 6-1 (1 g, 1.26 mmol, 1 eq) and N,N-dimethylformamide dimethyl acetal (116.61 mg, 5.06 mmol, 4 eq) were added to a flask containing methanol (10 mL). The reaction was stirred at 20 °C for 12 h. The reaction was monitored by LCMS and found to be complete. The reaction mixture was concentrated under reduced pressure. Ethyl acetate (10 mL) was added, and the mixture was washed three times with semi-saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 100:1 to 0:1, with 0.5% triethylamine) to obtain a white solid compound WX006 (0.8 g, yield: 78.7%, purity: 99.4%).

[0351] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 8.67 (s, 1H), 7.99 - 7.93 (m, 1H), 7.48 - 7.30 (m, 10H), 6.89 - 6.85 (m, 4H), 5.96 - 5.89 (m, 1H), 5.63 - 5.60 (m, 1H), 5.19 - 5.05 (m, 1H), 4.74 - 4.67 (m, 1H), 4.20 - 4.18 (m, 1H), 3.80 - 3.76 (m, 7H), 3.65 - 3.41 (m, 6H), 3.13 - 3.05 (m, 6H), 2.65 - 2.63 (m, 2H), 2.51 - 2.20 (m, 3H), 1.18 - 1.03 (m, 12H)

[0352] LCMS (ESI) m / z: 803.3 [M + H] +

[0353] Synthesis of Compound WX007 in Example 7

[0354] Synthetic Route:

[0355]

[0356] WX007 was synthesized by a method similar to that in Example 5.

[0357] Compound 7 - 1 (6 g, 7.48 mmol, 1 eq) and N,N - dimethylformamide dimethyl acetal (2.67 g, 22.44 mmol, 3 eq) were added to a flask containing methanol (60 mL). The reaction was stirred at 30 °C for 24 hours. LCMS was used to detect the completion of the reaction, and the reaction solution was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 100:1 to 0:1, adding 0.3% triethylamine) to obtain off - white solid compound WX007 (3.8 g, yield: 62.3%, purity: 97.0%).

[0358] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 8.65 (s, 1H), 8.11 - 7.97 (m, 1H), 7.37 - 7.30 (m, 9H), 6.89 - 6.85 (m, 4H), 5.95 - 5.92 (m, 1H), 5.61 - 5.58 (m, 1H), 4.55 - 4.42 (m, 1H), 4.17 - 4.15 (m, 1H), 3.89 - 3.40 (m, 17H), 3.11 - 3.03 (m, 6H), 2.65 - 2.31 (m, 2H), 1.18 - 1.09 (m, 12H)

[0359] LCMS(ESI) m / z: 815.5 [M+H] +

[0360] Synthesis of Compound WX008 in Example 8

[0361] Synthetic route:

[0362]

[0363] WX008 was synthesized by a method similar to that in Example 5.

[0364] Compound 8-1 (10 g, 12.95 mmol, 1 eq) and N,N-dimethylformamide dimethyl acetal (1.19 g, 51.82 mmol, 4 eq) were added to a flask containing methanol (100 mL). The reaction was stirred at 30 °C for 16 hours. The reaction was monitored by LCMS and found to be complete. The reaction mixture was concentrated under reduced pressure. Ethyl acetate (100 mL) was added, and the mixture was washed three times with semi-saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (dichloromethane:methanol = 1:0 to 10:1, with 0.5% triethylamine) to obtain off-white solid Compound WX008 (6.1 g, yield: 59.9%, purity: 98.2%).

[0365] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 8.64 (s, 1H), 7.85 - 7.80 (m, 1H), 7.44 - 7.29 (m, 10H), 6.88 - 6.84 (m, 4H), 6.21 - 6.17 (m, 1H), 5.73 - 5.71 (m, 1H), 4.63 - 4.56 (m, 1H), 4.12 - 4.09 (m, 1H), 3.76 - 3.57 (m, 11H), 3.37 - 3.32 (m, 2H), 3.12 - 3.04 (m, 6H), 2.28 - 2.20 (m, 2H), 1.15 - 0.97 (m, 12H)

[0366] LCMS(ESI) m / z: 785.3 [M+H] +

[0367] Synthesis of Compound WX009 in Example 9

[0368] Synthetic route:

[0369]

[0370] WX009 was synthesized by a method similar to that in Example 10.

[0371] Compound 9-1 (10 g, 10.30 mmol, 1 eq) and N,N-dimethylformamide dimethyl acetal (3.68 g, 30.92 mmol, 3.0 eq) were added to a flask containing methanol (100 mL). The reaction was stirred at 30 °C for 16 h. The reaction was monitored by LCMS and when it was complete, the reaction mixture was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 1:0 to 0:1, with 0.3% triethylamine) to obtain off-white solid compound WX009 (5 g, yield: 50.7%, purity: 99.7%).

[0372] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 9.90 (s, 1H), 8.47 - 8.46 (m, 1H), 7.79 - 7.77 (m, 1H), 7.46 - 7.21 (m, 9H), 6.84 - 6.81 (m, 4H), 5.94 - 5.93 (m, 1H), 4.81 - 4.78 (m, 1H), 4.40 - 4.26 (m, 2H), 3.80 - 3.75 (m, 8H), 3.62 - 3.59 (m, 3H), 3.36 - 3.34 (m, 2H), 3.03 - 3.00 (m, 6H), 2.66 - 2.40 (m, 2H), 1.17 - 1.00 (m, 12H), 0.80 - 0.79 (m, 9H), 0.01 - -0.10 (m, 6H)

[0373] LCMS (ESI) m / z: 953.4 [M-H] -

[0374] Synthesis of Compound WX010 in Example 10

[0375] Synthetic route:

[0376]

[0377] Compound 10-1 (45 g, 52.45 mmol, 1 eq) and N,N-dimethylformamide dimethyl acetal (9.38 g, 78.68 mmol, 1.5 eq) were added to a flask containing methanol (450 mL). The reaction was stirred at 25 °C for 3 h. The reaction was monitored by LCMS and when it was complete, the reaction mixture was concentrated under reduced pressure. The crude product was eluted by column chromatography (dichloromethane:methanol = 1:0 to 10:1, with 0.5% triethylamine) to obtain off-white solid compound WX010 (71 g, yield: 79%, purity: 98%).

[0378] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 10.38 (s, 1H), 8.49 (d, J = 4.0 Hz, 1H), 7.81 (s, 1H), 7.38 - 7.22 (m, 9H), 6.81–6.78 (m, 4H), 6.21–6.14 (m, 1H), 5.66–5.51 (m, 1H), 5.25–5.10 (m, 1H), 4.26–4.24 (m, 1H), 3.72–3.51 (m, 11H), 3.35–3.2 (m, 1H), 3.06 - 3.02 (m, 6H), 2.62 - 2.43 (m, 2H), 1.16–

[0379] 0.99 (m, 12H)

[0380] LCMS (ESI) m / z: 841.3 [M-H] -

[0381] Synthesis of Compound WX011 in Example 11

[0382] Synthetic Route:

[0383]

[0384] Compound WX011 was synthesized by a method similar to that in Example 10.

[0385] Compound 11-1 (10 g, 11.49 mmol, 1 eq) and N,N-dimethylformamide dimethyl acetal (4.11 g, 34.48 mmol, 3 eq) were added to a flask containing methanol (100 mL). The reaction was stirred at 20 °C for 12 hours. The reaction was monitored by LCMS and found to be complete. The reaction mixture was concentrated under reduced pressure. Ethyl acetate (100 mL) was added, and the mixture was washed 4 times with semi-saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (dichloromethane:methanol = 1:0 to 10:1, with 0.5% triethylamine) to obtain a white solid compound WX011 (8.7 g, yield: 88.5%, purity: 98.5%).

[0386] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 9.63 (s, 1H), 8.53 - 8.50 (m, 1H), 7.76 - 7.72 (m, 1H), 7.43 - 7.26 (m, 9H), 6.85 - 6.79 (m, 4H), 6.00 - 5.99 (m, 1H), 4.65 - 4.63 (m, 1H), 4.40 - 4.39 (m, 1H), 4.31 - 4.28 (m, 1H), 3.75 (s, 6H), 3.63 - 3.59 (m, 4H), 3.46 - 3.37 (m, 5H), 3.06 - 3.03 (m, 6H), 2.64 - 2.44 (m, 2H), 1.17 - 1.02 (m, 12H)

[0387] LCMS (ESI) m / z: 855.3 [M + H] +

[0388] Synthesis of Compound WX012 in Example 12

[0389] Synthetic Route:

[0390]

[0391] WX012 was synthesized by a method similar to that in Example 10.

[0392] Compound 12 - 1 (100 g, 119.06 mmol, 1 eq) and N,N - dimethylformamide dimethyl acetal (42.56 g, 357.17 mmol, 3 eq) were added to a flask containing methanol (1.0 L). The reaction was stirred at 35 °C for 12 hours. LCMS was used to detect the completion of the reaction, and the reaction solution was concentrated under reduced pressure. Ethyl acetate (1000 mL) was added, and the mixture was washed twice with semi - saturated brine (1000 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 1:0 to 0:1, with 0.3% triethylamine) to obtain a white solid compound WX012 (57 g, yield: 58%, purity: 99.1%).

[0393] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 10.22 (s, 1H), 8.51 (s, 1H), 7.73 (s, 1H), 7.40 - 7.22 (m, 9H), 6.81 - 6.77 (m, 4H), 6.30 - 6.28 (m, 1H), 4.82 - 4.77 (m, 1H), 4.17 - 4.05 (m, 1H), 3.75 (s, 6H), 3.64 - 3.63 (m, 3H), 3.30 - 3.25 (m, 2H), 3.05 - 3.01 (m, 6H), 2.63 - 2.58 (m, 1H), 2.53 - 2.50 (m, 3H), 1.22 - 1.05 (m, 12H)

[0394] LCMS (ESI) m / z: 825.5 [M+H] +

[0395] Synthesis of Compound WX013 in Example 13

[0396] Synthesis Route:

[0397]

[0398] Compound 1-2 (65 g, 73.52 mmol, 1 eq), Compound A (93.58 g, 294.09 mmol, 4 eq) and N,N-diisopropylethylamine (41.81 g, 323.50 mmol, 4.4 eq) were added to a flask containing dichloromethane (650 mL). The reaction was stirred at 20 °C for 22 hours. LCMS detected that the reaction was complete. The reaction solution was added to an aqueous solution of a mixture of sodium carbonate and sodium bicarbonate (400 mL), and then separated. The organic phase was washed with an aqueous solution of a mixture of sodium bicarbonate and sodium carbonate (300 mL × 10). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 250*70 mm #10um; mobile phase: acetonitrile; flow rate: 160 mL / min), and then concentrated to obtain a white solid compound WX013 (43 g, yield: 57%, purity: 99%).

[0399] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 9.50 (s, 1H), 8.56–8.53 (m, 1H), 8.32–8.31 (m, 1H), 7.47 - 7.45 (m, 2H), 7.34 - 7.31 (m, 9H), 7.01 - 7.00 (m, 2H), 6.83–6.81 (m, 4H), 6.03–6.01 (m, 1H), 5.14–5.11 (m, 1H), 4.98 (s, 1H), 4.36–4.35 (m, 2H), 3.73 - 3.70 (m, 2H), 3.65 (s, 6H), 3.63 - 3.35 (m, 4H), 2.67 - 2.44 (m, 2H), 1.19 - 1.07 (m, 12H), 0.76 (s, 9H), -0.01--0.20 (m, 6H)

[0400] LCMS (ESI) m / z: 1016.2 [M-H] -

[0401] Synthesis of Compound WX014 in Example 14

[0402] Synthesis Route:

[0403]

[0404] WX014 was synthesized by a method similar to that in Example 13.

[0405] Compound 2-2 (44 g, 57.00 mmol, 1 eq), Compound A (146 g, 456.60 mmol, 8 eq) and N,N-diisopropylethylamine (66.00 g, 510.66 mmol, 9 eq) were added to a flask containing dichloromethane (440 mL). The reaction was stirred at 30 °C for 10 hours. LCMS detected that the reaction was complete. The reaction solution was added to an aqueous solution of a mixture of sodium carbonate and sodium bicarbonate (500 mL), and then separated. The organic phase was washed with an aqueous solution of a mixture of sodium bicarbonate and sodium carbonate (300 mL × 10). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 180*70 mm #10um; mobile phase: acetonitrile; flow rate: 160 mL / min), and then concentrated to obtain a white solid compound WX014 (22 g, yield: 42.5%, purity: 99.5%).

[0406] 11H NMR (400 MHz, ACETONITRILE-d3) δ = 9.62 (s, 1H), 8.64 - 8.63 (m, 1H), 8.37 (s, 1H), 7.32 - 7.19 (m, 11H), 7.00 - 6.98 (m, 3H), 6.77 - 6.75 (m, 3H), 6.33 - 6.28 (m, 1H), 5.90 - 5.75 (m, 1H), 5.40 - 5.34 (m, 1H), 5.00 (s, 2H), 4.29 (s, 1H), 3.71 - 3.51 (m, 11H), 3.35 - 3.24 (m, 1H), 2.64 - 2.49 (m, 2H), 1.20 - 1.07 (m, 12H)

[0407] LCMS (ESI) m / z: 904.3 [M-H] -

[0408] Synthesis of Compound WX015 in Example 15

[0409] Synthetic Route:

[0410]

[0411] Compound 3-2 (68 g, 86.75 mmol, 1 eq), Compound A (96.61 g, 303.63 mmol, 3.5 eq) and N,N-diisopropylethylamine (49.33 g, 381.71 mmol, 4.4 eq) were added to a flask containing dichloromethane (680 mL). The reaction was stirred at 20 °C for 12 hours. LCMS detected that the reaction was complete. The reaction solution was added to an aqueous solution of a mixture of sodium carbonate and sodium bicarbonate (500 mL), and then separated. The organic phase was washed with an aqueous solution of a mixture of sodium bicarbonate and sodium carbonate (300 mL × 10). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 180*70 mm #10um; mobile phase: acetonitrile; flow rate: 160 mL / min), and then concentrated to obtain a white solid compound WX015 (36 g, yield: 45%, purity: 98%).

[0412] 11H NMR (400 MHz, ACETONITRILE-d3) δ = 9.47 (s, 1H), 8.47 (d, J = 7.2 Hz, 1H), 8.23 (d, J = 8.8 Hz, 1H), 7.33 - 7.25 (m, 2H), 7.22 - 7.02 (m, 9H), 6.92 - 6.84 (m, 3H), 6.70 - 6.67 (m, 4H), 6.02 (dd, J = 4.4 Hz, J = 2.4 Hz, 1H), 4.89 (s, 2H), 4.73 - 4.55 (m, 2H), 4.27 - 4.15 (m, 1H), 3.82 - 3.68 (m, 1H), 3.62 - 3.59 (m, 6H), 3.58 - 3.44 (m, 3H), 3.40 - 3.29 (m, 4H), 3.25 - 3.19 (m, 1H), 2.59 - 2.52 (m, 1H), 2.39 (t, J = 6.0 Hz, 1H), 1.12 - 0.96 (m, 12H)

[0413] LCMS (ESI) m / z: 916.2 [M-H] -

[0414] Synthesis of Compound WX016 in Example 16

[0415] Synthesis Route:

[0416]

[0417] Compound 4-2 (26 g, 34.49 mmol, 1 eq), Compound A (43.9 g, 137.96 mmol, 4 eq) and N,N-diisopropylethylamine (19.61 g, 151.76 mmol, 4.4 eq) were added to a flask containing dichloromethane (260 mL). The reaction was stirred at 20 °C for 12 hours. LCMS detected that the reaction was complete. The reaction solution was separated by liquid-liquid extraction with an aqueous solution of sodium carbonate and sodium bicarbonate (200 mL). The organic phase was washed with an aqueous solution of sodium bicarbonate and sodium carbonate (120 mL × 10), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 3:1 to 0:1, adding 0.5% triethylamine) to obtain off-white solid Compound WX016 (26 g, yield: 85%, purity: 98%).

[0418] 11H NMR (400 MHz, ACETONITRILE-d3) δ = 9.76 (s, 1H), 8.56 (s, 1H), 8.33 (s, 1H), 7.35 - 7.27 (m, 2H), 7.25 - 7.22 (m, 9H), 6.98 - 6.96 (m, 3H), 6.76 - 6.74 (m, 4H), 6.43 - 6.40 (m, 1H), 5.02 (s, 2H), 4.94 - 4.88 (m, 1H), 4.26 - 4.24 (m, 1H), 3.73 - 3.59 (m, 10H), 3.45 - 3.20 (m, 2H), 3.15 - 3.05 (m, 1H), 2.65–2.52 (m, 3H), 1.23–1.09 (m, 12H)

[0419] LCMS (ESI) m / z: 886.2 [M-H] -

[0420] Synthesis of Compound WX017 in Example 17

[0421] Synthesis Route:

[0422]

[0423] Step 1:

[0424] Compound 5-1 (60 g, 66.51 mmol, 1 eq) and triethylamine (67.30 g, 665.12 mmol, 10 eq) were added to a flask containing methanol (245 mL). The reaction was stirred at 25 °C for 12 hours. LCMS detected that the reaction was complete. The reaction solution was concentrated under reduced pressure. The crude product was dissolved in methyl tert-butyl ether (120 mL) and added dropwise to n-hexane (1200 mL). The mixture was stirred at room temperature for 0.5 hour, filtered, and the filter cake was concentrated under reduced pressure to obtain a white solid compound 17-1 (55 g, yield: 95%, purity: 99%).

[0425] Step 2:

[0426] Compound 17-1 (72 g, 83.72 mmol, 1 eq), compound C (31.16 g, 108.83 mmol, 1.3 eq) and triethylamine (16.94 g, 167.43 mmol, 2 eq) were added to a flask containing dichloromethane (360 mL). The reaction was stirred at 25 °C for 1 hour. The reaction was monitored by LCMS and found to be complete. The reaction mixture was added to an aqueous sodium bicarbonate solution (300 mL), and then separated by liquid-liquid extraction. The organic phase was washed with an aqueous sodium bicarbonate solution (300 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was dissolved in methyl tert-butyl ether (300 mL) and purified by column chromatography. The resulting organic phase was concentrated to obtain a pale yellow solid compound WX017 (79 g, yield: 95%, purity: 99%).

[0427] 1H NMR (400 MHz, ACETONITRILE-d3) δ=10.25 (s, 1H), 8.49–8.42 (m, 1H), 7.50–7.39 (m, 2H), 7.38-7.30 (m, 9H), 7.28–6.88 (m, 8H), 5.70–5.62 (m, 1H), 4.79–4.65 (m, 2H), 4.39–4.30 (m, 3H), 3.74 (s, 6H), 3.63-3.50 (m, 6H), 2.59-2.46 (m, 2H), 1.14–0.91 (m, 12H), 0.90 (s, 9H), 0.21-0.13 (m, 6H)

[0428] LCMS (ESI) m / z: 992.4 [M-H] -

[0429] Example 18: Synthesis of Compound WX018

[0430] Synthesis route:

[0431]

[0432] Step 1:

[0433] Compound 6-1 (39 g, 49.38 mmol, 1 eq) and triethylamine (49.97 g, 493.78 mmol, 10 eq) were added to a flask containing methanol (200 mL). The reaction was stirred at 25 °C for 12 hours. The reaction was monitored by LCMS and found to be complete. The reaction mixture was concentrated under reduced pressure. The crude product was dissolved in methyl tert-butyl ether (80 mL) and added dropwise to n-hexane (800 mL). The mixture was stirred at room temperature for 0.5 hour, filtered, and the filter cake was concentrated under reduced pressure to obtain an off-white solid compound 18-1 (41 g, yield: 99%, purity: 99%).

[0434] Step 2:

[0435] Compound 18-1 (24 g, 28.89 mmol, 1 eq), compound C (10.75 g, 37.55 mmol, 1.3 eq) and triethylamine (7.6 g, 75.1 mmol, 2.6 eq) were added to a flask containing dichloromethane (120 mL). The reaction was stirred at 25 °C for 1 hour. The reaction was detected to be complete by LCMS. The reaction solution was added to an aqueous sodium bicarbonate solution (100 mL), and then separated by liquid-liquid extraction. The organic phase was washed with an aqueous sodium bicarbonate solution (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 250*70mm #10um; mobile phase: acetonitrile; flow rate: 90 mL / min), and then concentrated to obtain a white solid compound WX018 (20 g, yield: 71%, purity: 99%).

[0436] 1H NMR (400 MHz, ACETONITRILE-d3) δ=9.49 (s, 1H), 8.43–8.37 (m, 1H), 7.48–7.30 (m, 11H), 6.99–6.86 (m, 7H), 5.95–5.89 (m, 1H), 5.23–5.09 (m, 1H), 4.72–4.71 (m, 3H), 4.24–4.22 (m, 1H), 3.76 - 3.56 (m, 12H), 2.61 - 2.48 (m, 2H), 1.18–1.02 (m, 12H)

[0437] LCMS (ESI) m / z: 880.2 [M-H] -

[0438] Synthesis of Compound WX019 in Example 19

[0439] Synthetic route:

[0440]

[0441] WX019 was synthesized by a method similar to that in Example 17.

[0442] Step 1:

[0443] Compound 7-1 (70 g, 87.30 mmol, 1 eq) and triethylamine (88.33 g, 872.97 mmol, 10 eq) were added to a flask containing methanol (700 mL). The reaction was stirred at 25 °C for 12 hours. The reaction was detected to be complete by LCMS. The reaction solution was concentrated under reduced pressure. The crude product was dissolved in methyl tert-butyl ether (490 mL) and added dropwise to n-hexane (70 mL). The mixture was stirred at room temperature for 2 hours, filtered, and the filter cake was concentrated under reduced pressure to obtain a white solid compound 19-1 (65 g, yield: 96.8%, purity: 98.8%).

[0444] Step 2:

[0445] Compound 19-1 (52 g, 68.44 mmol, 1 eq), compound C (19.59 g, 68.44 mmol, 1.0 eq) and triethylamine (13.85 g, 136.87 mmol, 2 eq) were added to a flask containing dichloromethane (260 mL). The reaction was stirred at 25 °C for 1 hour. The reaction was detected to be complete by LCMS. The reaction solution was added to an aqueous sodium bicarbonate solution (260 mL) and then separated. The organic phase was washed with an aqueous sodium bicarbonate solution (260 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 100:1 to 0:1, adding 0.5% triethylamine) to obtain a white solid compound WX019 (55 g, yield: 88.9%, purity: 98.9%).

[0446] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 9.83 (s, 1H), 8.55 - 8.44 (m, 1H), 7.49 - 7.30 (m, 11H), 6.99 - 6.86 (m, 8H), 5.85 - 5.80 (m, 1H), 4.76 - 4.75 (m, 2H), 4.60 - 4.44 (m, 1H), 4.18 - 4.15 (m, 1H), 3.89 - 3.88 (m, 1H), 3.75 - 3.71 (m, 8H), 3.62 - 3.42 (m, 8H), 2.64 - 2.50 (m, 2H), 1.20 - 1.01 (m, 12H)

[0447] LCMS (ESI) m / z: 892.2 [M-H] -

[0448] Synthesis of Compound WX020 in Example 20

[0449] Synthesis route:

[0450]

[0451] Synthesize WX020 using a method similar to that of Example 17.

[0452] Step 1:

[0453] Add compound 20-1 (70 g, 83.94 mmol, 1 eq) to a flask containing amine methanol (7 M, 700 mL). The reaction was stirred at 20 °C for 16 hours. The reaction was monitored by LCMS and found to be complete. The reaction mixture was concentrated under reduced pressure. The crude product was purified by column chromatography (dichloromethane:acetone = 100:0 to 8:1, with 0.5% triethylamine) to give a white solid compound 20-2 (40 g, yield: 65.2%, purity: 97.4%).

[0454] Step 2:

[0455] Add compound 20-2 (20.9 g, 27.46 mmol, 1 eq), compound C (8.64 g, 30.2 mmol, 1.1 eq) and triethylamine (5.5 g, 57.27 mmol, 2 eq) to a flask containing dichloromethane (105 mL). The reaction was stirred at 25 °C for 16 hours. The reaction was monitored by LCMS and found to be complete. The reaction mixture was added to an aqueous sodium bicarbonate solution (105 mL), and the layers were separated. The organic layer was washed with an aqueous sodium bicarbonate solution (105 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (dichloromethane:methanol = 100:1 to 0:1, with 0.5% triethylamine) to give a white solid compound WX020 (22.5 g, yield: 50%, purity: 98.8%).

[0456] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 9.29 (s, 1H), 8.27 - 8.20 (m, 1H), 7.43 - 7.00 (m, 11H), 6.88 - 6.85 (m, 8H), 6.11 - 6.06 (m, 1H), 4.70 (s, 2H), 4.63 - 4.56 (m, 1H), 4.15 - 4.13 (m, 1H), 3.76 - 3.50 (m, 10H), 3.40 - 3.36 (m, 2H), 2.63 - 2.51 (m, 3H), 2.31 - 2.29 (m, 1H), 1.17 - 1.06 (m, 12H)

[0457] LCMS (ESI) m / z: 862.4 [M-H] -

[0458] Synthesis of Compound WX021 in Example 21

[0459] Synthetic route:

[0460]

[0461] Compound 9-1 (95 g, 97.92 mmol, 1 eq) and amine in methanol (7 M, 475 mL) were added to a flask containing methanol (475 mL). The reaction was stirred at 25 °C for 16 hours. The reaction was monitored by LCMS and found to be complete. The reaction mixture was filtered, and the filter cake was concentrated under reduced pressure to obtain off-white solid Compound 21-1 (80 g, yield: 90%, purity: 99%).

[0462] Step 2:

[0463] Compound 21-1 (65 g, 72.22 mmol, 1 eq), Compound IV-1 (79.7 g, 252.75 mmol, 3.5 eq) and N,N-diisopropylethylamine (42 g, 324.97 mmol, 4.5 eq) were added to a flask containing dichloromethane (455 mL). The reaction was stirred at 25 °C for 64 hours. The reaction was monitored by LCMS and found to be complete. The reaction mixture was separated by liquid-liquid extraction with an aqueous mixture of sodium carbonate and sodium bicarbonate (400 mL). The organic layer was washed with an aqueous mixture of sodium bicarbonate and sodium carbonate (400 mL × 10), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 1:0 to 0:1, with 0.5% triethylamine) to obtain off-white solid Compound WX021 (70 g, yield: 87%, purity: 97%).

[0464] 1 H NMR (400 MHz, ACETONITRILE-d3) δ = 7.97 (s, 1H), 7.47 - 7.46 (m, 2H), 7.35 - 7.18 (m, 9H), 6.89 - 6.80 (m, 6H), 5.91–5.87 (m, 1H), 4.89 - 4.86 (m, 1H), 4.67 - 4.65 (m, 2H), 4.40 - 4.29 (m, 2H), 3.61 - 3.58 (m, 10H), 3.44 - 3.42 (m, 2H), 2.88–2.83 (m, 1H), 2.66–2.39 (m, 2H), 1.20–1.02 (m, 18H), 0.78 (s, 9H), 0.00--0.13 (m, 6H)

[0465] LCMS (ESI) m / z: 1074.3 [M-H] -

[0466] Synthesis of Compound WX022 in Example 22

[0467] Synthetic route:

[0468]

[0469] Step 1:

[0470] Compound 10-1 (33 g, 38.47 mmol, 1 eq) and amine in methanol (7 M, 66 mL) were added to a flask containing methanol (240 mL). The reaction was stirred at 25 °C for 40 hours. The reaction was monitored by LCMS and was found to be complete. The reaction mixture was concentrated under reduced pressure to obtain crude compound 22-1 (35 g) as an off-white solid.

[0471] Step 2

[0472] Compound 22-1 (29 g, 36.81 mmol, 1 eq), compound IV-1 (40.63 g, 128.84 mmol, 3.5 eq) and N,N-diisopropylethylamine (21.41 g, 165.65 mmol, 4.5 eq) were added to a flask containing dichloromethane (200 mL). The reaction was stirred at 25 °C for 20 hours. The reaction was monitored by LCMS and was found to be complete. The reaction mixture was separated by liquid-liquid extraction with an aqueous mixture of sodium carbonate and sodium bicarbonate (200 mL). The organic layer was washed with an aqueous mixture of sodium bicarbonate and sodium carbonate (200 mL × 10), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 250*100mm #10um; mobile phase: acetonitrile; flow rate: 200 mL / min) and then concentrated to obtain off-white solid compound WX022 (10 g, yield: 33%, purity: 99%).

[0473] 1 H NMR (400 MHz, ACETONITRILE-d3) δ = 7.96 (s, 1H), 7.42 - 7.30 (m, 2H), 7.27 - 7.21 (m, 9H), 6.96 - 6.80 (m, 6H), 6.20 – 6.15 (m, 1H), 5.62 – 5.48 (m, 1H), 4.95 - 4.75 (m, 3H), 4.33 - 4.30 (m, 1H), 3.74 - 3.44 (m, 13H), 2.90 – 2.89 (m, 1H), 2.51 – 2.47 (m, 2H), 1.23 – 1.05 (m, 19H)

[0474] LCMS (ESI) m / z: 962.2 [M-H] -

[0475] Synthesis of Compound WX023 in Example 23

[0476] Synthetic route:

[0477]

[0478] Compound 11-1 (50 g, 57.48 mmol, 1 eq) was added to a flask containing ethylamine (2 M solution in tetrahydrofuran, 500 mL), and the reaction was stirred at 25 °C for 60 h. The reaction was monitored by LCMS and was found to be complete. The reaction mixture was concentrated under reduced pressure. The crude product was dissolved in ethyl acetate (110 mL) and slowly added dropwise to n-heptane (770 mL). The mixture was stirred for 15 min, filtered, and the filter cake was concentrated under reduced pressure to give a white solid, compound 23-1 (45 g, yield: 98%, purity: 98%).

[0479] Step 2:

[0480] Compound 23-1 (45 g, 56.26 mmol, 1 eq), compound IV-1 (62.09 g, 196.91 mmol, 3.5 eq) and N,N-diisopropylethylamine (32.72 g, 253.17 mmol, 4.5 eq) were added to a flask containing dichloromethane (450 mL), and the reaction was stirred at 25 °C for 20 h. The reaction was monitored by LCMS and was found to be complete. The reaction mixture was separated by adding an aqueous solution of sodium carbonate and sodium bicarbonate (200 mL). The organic layer was washed with an aqueous solution of sodium bicarbonate and sodium carbonate (200 mL × 10), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 250*100mm#10um; mobile phase: acetonitrile; flow rate: 200 mL / min) and then concentrated to give a white solid, compound WX023 (20 g, yield: 36%, purity: 99%).

[0481] 1 H NMR (400 MHz, ACETONITRILE-d3) δ=7.96–7.94 (m, 1H), 7.45-7.43 (m, 2H), 7.31-7.17 (m, 10H), 6.82-6.79 (m, 6H), 5.97–5.94 (m, 1H), 4.73–4.71 (m, 2H), 4.59-4.57 (m, 2H), 4.43-4.31 (m, 1H), 3.72-3.59 (m, 10H), 3.44–3.41 (m, 6H), 2.85–2.83 (m, 1H), 2.64–2.45 (m, 2H), 1.20–1.06 (m, 19H)

[0482] LCMS (ESI) m / z: 974.2 [M-H] -

[0483] Synthesis of Compound WX024 in Example 24

[0484] Synthetic route:

[0485]

[0486] WX024 was synthesized by a method similar to that of Example 21.

[0487] Step 1:

[0488] Compound 12-1 (4 g, 4.76 mmol, 1 eq) was added to a flask containing amine methanol (7 M, 40 mL), and the reaction was stirred at 20 °C for 16 h. The reaction was monitored by LCMS and was found to be complete. The reaction mixture was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 1:0 to 0:1, with 0.5% triethylamine) to obtain a white solid compound 24-1 (2.9 g, yield: 79.1%, purity: 97.9%).

[0489] Step 2:

[0490] Compound 24-1 (2.9 g, 36.81 mmol, 1 eq), compound IV-1 (4.75 g, 15.06 mmol, 4 eq) and N,N-diisopropylethylamine (2.43 g, 18.83 mmol, 5 eq) were added to a flask containing dichloromethane (29 mL), and the reaction was stirred at 25 °C for 64 h. The reaction was monitored by TLC and was found to be complete. The reaction mixture was separated by adding an aqueous solution of sodium carbonate and sodium bicarbonate (50 mL), and the organic phase was washed with an aqueous solution of sodium bicarbonate and sodium carbonate (50 mL × 10). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 1:0 to 0:1, with 0.5% triethylamine) to obtain a white solid compound WX024 (2.2 g, yield: 61.7%, purity: 99.0%).

[0491] 1 H NMR (400 MHz, ACETONITRILE-d3) δ = 7.89 (s, 1H), 7.37 - 7.27 (m, 2H), 7.25 - 7.17 (m, 9H), 6.93 - 6.80 (m, 2H), 6.76 - 6.75 (m, 4H), 6.26 - 6.23 (m, 1H), 4.78 - 4.70 (m, 3H), 4.23 - 4.22 (m, 1H), 3.70 - 3.56 (m, 10H), 3.31 - 3.28 (m, 2H), 2.87 - 2.83 (m, 2H), 2.63 - 2.51 (m, 3H), 1.20 - 1.07 (m, 18H)

[0492] Synthesis of Compound WX025 in Example 25

[0493] Synthesis route:

[0494]

[0495] Synthesize WX025 using a method similar to that of Example 5.

[0496] Step 1:

[0497] Add compound 25-1 (10 g, 10.26 mmol, 1 eq) and N,N-dimethylformamide dimethyl acetal (3.06 g, 25.66 mmol, 2.5 eq) to a flask containing methanol (100 mL). Stir the reaction at 30 °C for 24 hours. Detect the completion of the reaction by LCMS and concentrate the reaction solution under reduced pressure. Add ethyl acetate (100 mL) and wash it 3 times with semi-saturated brine (100 mL). Dry the organic phase with anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure. Elute the crude product by column chromatography (petroleum ether:ethyl acetate = 3:1 to 0:1, adding 1% triethylamine) to obtain off-white solid compound WX025 (5.9 g, yield: 57.8%, purity: 98.0%).

[0498] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 8.65 (s, 1H), 7.92 - 7.83 (m, 1H), 7.48 - 7.31 (m, 9H), 6.88 - 6.85 (m, 4H), 6.05 - 6.04 (m, 1H), 5.71 - 5.69 (m, 1H), 5.10 - 5.02 (m, 2H), 4.40 - 4.37 (m, 3H), 3.76 - 3.38 (m, 13H), 3.13 - 3.05 (m, 6H), 2.65 - 2.46 (m, 2H), 1.16 - 1.13 (m, 12H), 1.09 - 1.00 (m, 20H)

[0499] Synthesis of Compound WX026 in Example 26

[0500] Synthesis route:

[0501]

[0502] Synthesize WX026 using a method similar to that of Example 2.

[0503] Step 1:

[0504] Compound 26-1 (10 g, 10.02 mmol, 1 eq) and N,N-dimethylformamide dimethyl acetal (4.77 g, 40.07 mmol, 4 eq) were added to a flask containing methanol (70 mL). The reaction was stirred at 30 °C for 12 hours. The reaction was monitored by LCMS and was found to be complete. The reaction mixture was concentrated under reduced pressure. Methyl tert-butyl ether (100 mL) was added, and the mixture was washed three times with semi-saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 1:1 to 0:1, with 0.3% triethylamine) to obtain a white solid compound WX026 (6 g, yield: 58.5%, purity: 98.8%).

[0505] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 8.88 (s, 1H), 8.34 - 8.31 (m, 1H), 8.12 - 8.10 (m, 1H), 7.41 - 7.22 (m, 9H), 6.82 - 6.78 (m, 4H), 6.14 - 6.12 (m, 1H), 5.31 - 5.26 (m, 1H), 5.03 - 5.01 (m, 2H), 4.99 - 4.94 (m, 1H), 4.34 - 4.33 (m, 1H), 3.73 - 3.31 (m, 12H), 3.13 - 3.05 (m, 6H), 2.69 - 2.49 (m, 2H), 1.20 - 1.11 (m, 12H), 0.90 - 0.86 (m, 20H)

[0506] LCMS (ESI) m / z: 1011.5 [M+H] +

[0507] Example 27 Synthesis of Compound WX027

[0508] Synthesis route:

[0509]

[0510] WX027 was synthesized by a method similar to that of Example 10.

[0511] Step 1:

[0512] Compound 27-1 (10 g, 9.86 mmol, 1 eq) and N,N-dimethylformamide dimethyl acetal (5.29 g, 44.37 mmol, 4.5 eq) were added to a flask containing methanol (100 mL). The reaction was stirred at 30 °C for 16 hours. The reaction was monitored by LCMS and was found to be complete. The reaction mixture was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 250*70 mm #10um; mobile phase: acetonitrile; flow rate: 200 mL / min) and then concentrated to obtain a white solid compound WX027 (6.5 g, yield: 64.1%, purity: 97.4%).

[0513] 1H NMR (400 MHz, ACETONITRILE-d3) δ=9.80 (s, 1H), 8.47 (s, 1H), 7.74 (s, 1H), 7.42 - 7.24 (m, 9H), 6.83 - 6.80 (m, 4H), 6.06 - 6.04 (m, 1H), 5.00 - 4.98 (m, 1H), 4.93 - 4.90 (m, 2H), 4.51 - 4.50 (m, 1H), 4.26 - 4.25 (m, 1H), 3.74–3.73 (m, 6H), 3.65 - 3.59 (m, 4H), 3.58 - 3.40 (m, 2H), 3.03 (s, 6H), 2.44 - 2.41 (m, 2H), 1.16 - 1.14 (m, 12H), 0.93 - 0.89 (m, 21H)

[0514] LCMS (ESI) m / z: 1025.6 [M+H] +

[0515] Synthesis of Compound WX028 in Example 28

[0516] Synthetic route:

[0517]

[0518] WX028 was synthesized by a method similar to that in Example 17.

[0519] Step 1:

[0520] Compound 25-1 (13 g, 13.34 mmol, 1 eq) and triethylamine (13.50 g, 133.44 mmol, 10 eq) were added to a flask containing methanol (50 mL). The reaction was stirred at 30 °C for 16 h. The reaction was monitored by LCMS and found to be complete. The reaction mixture was concentrated under reduced pressure. The crude product was dissolved in methyl tert-butyl ether (130 mL) and added dropwise to n-hexane (1300 mL). The mixture was stirred at room temperature for 0.5 h, filtered, and the filter cake was concentrated under reduced pressure to obtain a white solid, compound 28-1 (11 g, yield: 88.4%, purity: 97.8%).

[0521] Step 2:

[0522] Compound 28-1 (10.6 g, 11.37 mmol, 1 eq), compound C (4.23 g, 14.77 mmol, 1.3 eq) and triethylamine (2.3 g, 22.74 mmol, 2 eq) were added to a flask containing dichloromethane (20 mL). The reaction was stirred at 25 °C for 1 h. The reaction was monitored by LCMS and found to be complete. The reaction mixture was added to an aqueous sodium bicarbonate solution (100 mL) and then separated. The organic phase was washed with an aqueous sodium bicarbonate solution (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 250*70mm#10um; mobile phase: acetonitrile; flow rate: 200 mL / min) and then concentrated to obtain a white solid, compound WX028 (4.7 g, yield: 64.1%, purity: 98.0%).

[0523] 1H NMR (400 MHz, ACETONITRILE-d3) δ=9.59 (s, 1H), 8.43 - 8.32 (m, 1H), 7.49 - 7.47 (m, 2H), 7.37 - 7.31 (m, 9H), 6.99 - 6.86 (m, 8H), 5.98 (s, 1H), 5.17 - 5.10 (m, 1H), 4.74 - 4.72 (m, 2H), 4.44 - 4.23 (m, 3H), 3.75 - 3.69 (m, 7H), 3.57 - 3.42 (m, 6H), 2.62 - 2.48 (m, 2H), 1.16 - 1.11 (m, 12H), 1.06 - 1.01 (m, 21H)

[0524] LCMS (ESI) m / z: 1064.5 [M-H] -

[0525] Synthesis of Compound WX029 in Example 29

[0526] Synthesis route:

[0527]

[0528] WX029 was synthesized by a method similar to that of Example 13.

[0529] Step 1:

[0530] Compound 26-1 (15 g, 15.02 mmol, 1 eq) was added to a flask containing ethylamine (2 M solution in tetrahydrofuran, 150 mL). The reaction was stirred at 15 °C for 12 h. The reaction was monitored by LCMS and when it was complete, the reaction mixture was concentrated under reduced pressure. Methyl tert-butyl ether (100 mL) was added, and the mixture was washed three times with semi-saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 4:1 to 0:1, with 0.3% triethylamine) to obtain a off-white solid compound 29-1 (11.7 g, yield: 81.4%, purity: 100%).

[0531] Step 2:

[0532] Compound 29-1 (8.5 g, 8.88 mmol, 1 eq), compound A (22.6 g, 71.02 mmol, 8 eq) and N,N-diisopropylethylamine (10.34 g, 80.00 mmol, 10 eq) were added to a flask containing dichloromethane (85 mL). The reaction was stirred at 30 °C for 12 h. The reaction was monitored by LCMS and when it was complete, the reaction mixture was separated by adding an aqueous solution of sodium carbonate and sodium bicarbonate (120 mL). The organic phase was washed with an aqueous solution of sodium bicarbonate and sodium carbonate (120 mL × 10), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 3:1 to 0:1, with 0.3% triethylamine) to obtain a off-white solid compound WX029 (5 g, yield: 51%, purity: 98.4%).

[0533] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 9.63 (s, 1H), 8.54 - 8.52 (m, 1H), 8.34 - 8.32 (m, 1H), 7.41 - 7.32 (m, 2H), 7.30 - 7.24 (m, 9H), 7.00 - 6.98 (m, 3H), 6.82 - 6.78 (m, 4H), 6.18 - 6.16 (m, 1H), 5.27 - 5.25 (m, 1H), 5.02 - 4.92 (m, 4H), 4.85 - 4.67 (m, 1H), 4.39 - 4.38 (m, 1H), 3.73 - 3.67 (m, 7H), 3.66 - 3.35 (m, 5H), 2.68 - 2.49 (m, 2H), 1.20 - 1.10 (m, 12H), 0.86 - 0.81 (m, 21H)

[0534] LCMS(ESI) m / z: 1088.6 [M-H] -

[0535] Synthesis of Compound WX030 in Example 30

[0536] Synthesis Route:

[0537]

[0538] WX030 was synthesized by a method similar to that in Example 21.

[0539] Step 1:

[0540] Compound 27-1 (20 g, 19.72 mmol, 1 eq) was added to a flask containing ethylamine (2 M solution in tetrahydrofuran, 207 mL). The reaction was stirred at 20 °C for 36 hours. The reaction was monitored by LCMS and when it was complete, the reaction mixture was concentrated under reduced pressure. Methyl tert-butyl ether (150 mL) was added, stirred for 15 minutes, filtered, and the filter cake was concentrated under reduced pressure to obtain a pale white crude solid compound 30-1 (23 g). It was directly used for the next step.

[0541] Step 2:

[0542] Compound 30-1 (13 g, 13.37 mmol, 1 eq), compound IV-1 (14.76 g, 46.80 mmol, 3.5 eq) and N,N-diisopropylethylamine (7.78 g, 60.17 mmol, 4.5 eq) were added to a flask containing dichloromethane (91 mL). The reaction was stirred at 15 °C for 120 hours. The reaction was monitored by LCMS and when it was complete, the reaction mixture was separated by liquid-liquid extraction with an aqueous mixture of sodium carbonate and sodium bicarbonate (100 mL). The organic phase was washed with an aqueous mixture of sodium bicarbonate and sodium carbonate (100 mL × 10), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 10:1 to 0:1, with 0.3% triethylamine) to obtain a pale yellow solid compound WX030 (7.9 g, yield: 50.4%, purity: 98.0%).

[0543] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 11.77 (s, 1H), 8.15 (s, 1H), 7.38 - 7.36 (m, 2H), 7.28 - 7.14 (m, 10H), 6.90 - 6.82 (m, 6H), 5.99 - 5.98 (m, 1H), 4.96 - 4.92 (m, 2H), 4.87 - 4.84 (m, 3H), 4.45 - 4.25 (m, 2H), 3.72 (s, 6H), 3.66 - 3.55 (m, 5H), 2.82 - 2.80 (m, 1H), 2.61 - 2.50 (m, 2H), 1.21 - 1.12 (m, 18H), 0.90 - 0.76 (m, 21H)

[0544] LCMS (ESI) m / z: 1146.6 [M - H] -

[0545] Synthesis of Compound WX031 in Example 31

[0546] Synthesis Route:

[0547]

[0548] Step 1:

[0549] Compound 4 - 2 (2 g, 2.65 mmol, 1 eq) and N,N - diethylformamide dimethyl acetal (2.34 g, 15.91 mmol, 6 eq) were added to a flask containing dichloromethane (20 mL). The reaction was stirred at 15 °C for 12 hours. LCMS was used to detect the completion of the reaction, and the reaction solution was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 250*70 mm #10um; mobile phase: acetonitrile; flow rate: 100 mL / min) and then concentrated to obtain a white solid compound WX031 (1.8 g, yield: 81.0%, purity: 98.7%).

[0550] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 8.94 (s, 1H), 8.38 (s, 1H), 8.11 (s, 1H), 7.39 - 7.30 (m, 2H), 7.27 – 7.24 (m, 7H), 6.80 - 6.77 (m, 4H), 6.44 - 6.41 (m, 1H), 4.94 - 4.90 (m, 1H), 4.23 - 4.21 (m, 1H), 3.76 - 3.48 (m, 14H), 3.47 - 3.25 (m, 2H), 3.20 – 3.05 (m, 1H), 2.67 - 2.55 (m, 3H), 1.29 - 1.17 (m, 18H)

[0551] LCMS(ESI) m / z: 837.4 [M+H] +

[0552] Synthesis of Compound WX032 in Example 32

[0553] Synthesis route:

[0554]

[0555] Step 1:

[0556] Compound 4-2 (2 g, 2.65 mmol, 1 eq) and N,N-dimethylacetamide dimethyl acetal (2.12 g, 15.91 mmol, 6 eq) were added to a flask containing pyridine (20 mL). The reaction was stirred at 35 °C for 72 hours. LCMS was used to detect the completion of the reaction, and the reaction solution was concentrated under reduced pressure. The crude product was purified by machine fractionation (column: Welch Xtimate C18 250*70 mm #10um; mobile phase: acetonitrile; flow rate: 100 mL / min), and then eluted by column chromatography (petroleum ether:acetone = 10:1 to 3:1, adding 0.3% triethylamine) to obtain a white solid compound WX032 (0.4 g, yield: 18.32%, purity: 95.0%).

[0557] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 8.38 (s, 1H), 8.06 (s, 1H), 7.36 - 7.26 (m, 2H), 7.24–7.22 (m, 7H), 6.79 - 6.38 (m, 4H), 6.41 - 6.38 (m, 1H), 4.92 - 4.88 (m, 1H), 4.20 - 4.19 (m, 1H), 3.73 - 3.58 (m, 10H), 3.40 - 3.20 (m, 2H), 3.19–3.10 (m, 7H), 2.70 - 2.52 (m, 3H), 2.00 (s, 3H), 1.18 - 1.08 (m, 12H) LCMS(ESI) m / z: 823.1 [M+H] +

[0558] Synthesis of Compound WX033 in Example 33

[0559] Synthesis route:

[0560]

[0561] Step 1:

[0562] Compound 4-2 (2 g, 2.65 mmol, 1 eq) and N,N-dibutylformamide dimethyl acetal (4.04 g, 15.91 mmol, 6 eq) were added to a flask containing dichloromethane (20 mL). The reaction was stirred at 15 °C for 12 hours. The reaction was monitored by LCMS and found to be complete. The reaction mixture was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 250*70 mm #10um; mobile phase: acetonitrile; flow rate: 120 mL / min), concentrated, and a white solid compound WX033 (1.6 g, yield: 67.5%, purity: 96.9%) was obtained.

[0563] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 8.95 (s, 1H), 8.38 (s, 1H), 8.12 (s, 1H), 7.39 - 7.26 (m, 2H), 7.25 – 7.21 (m, 7H), 6.81 - 6.77 (m, 4H), 6.45 - 6.42 (m, 1H), 4.96 - 4.90 (m, 1H), 4.23 - 4.20 (m, 1H), 3.84 - 3.61 (m, 12H), 3.43 - 3.02 (m, 6H), 2.68 - 2.55 (m, 3H), 1.67 – 1.57 (m, 4H), 1.40 - 1.10 (m, 18H), 0.97 – 0.93 (m, 7H)

[0564] LCMS (ESI) m / z: 893.4 [M+H] +

[0565] Synthesis of Compound WX034 in Example 34

[0566] Synthetic route:

[0567]

[0568] Step 1:

[0569] Compound 4-2 (2 g, 2.65 mmol, 1 eq) and 2,2-dimethoxy-1-methylpyrrolidine (2.31 g, 15.91 mmol, 6 eq) were added to a flask containing dichloromethane (20 mL). The reaction was stirred at 15 °C for 1 hour. The reaction was monitored by LCMS and found to be complete. The reaction mixture was concentrated under reduced pressure. The crude product was eluted by column chromatography (petroleum ether:ethyl acetate = 1:3 to 0:1, adding 0.3% triethylamine), and a white solid compound WX034 (1.0 g, yield: 45.1%, purity: 99.5%) was obtained.

[0570] 1H NMR (400 MHz, ACETONITRILE-d3) δ = 8.31 (s, 1H), 8.02 (s, 1H), 7.33 - 7.31 (m, 2H), 7.24–7.15 (m, 7H), 6.74 - 6.70 (m, 4H), 6.37 - 6.34 (m, 1H), 4.90–4.86 (m, 1H), 4.15 - 4.12 (m, 1H), 4.00–3.42 (m, 10H), 3.41 - 3.39 (m, 2H), 3.37 - 3.25 (m, 3H), 3.09–3.01 (m, 4H), 2.80 - 2.73 (m, 2H), 2.67–2.48 (m, 3H), 1.93–1.88 (m, 2H), 1.13–1.03 (m, 12H)

[0571] LCMS (ESI) m / z: 835.3 [M + H] +

[0572] Screening Examples 1 - 15

[0573]

[0574] Using an operation similar to that in Step 2 of Example 21, the feed amount of Compound 21 - 1 was 0.1 g, and the reaction was monitored by HPLC. The screening of reaction conditions was as follows:

[0575]

[0576] Among them, 10V means that the volume (mL) of the reagent used is 10 times the gram weight of the raw material. For example, in Screening Example 1, the feed amount of Compound 21 - 1 was 0.1 g, and 10V indicates that the amount of DCM used was 1 mL.

[0577] Screening Examples 16 - 21

[0578]

[0579] The feed amount of Compound 21 - 1 was 0.05 g, and the reaction was monitored by HPLC. The screening of the condensation reaction conditions was as follows:

[0580]

[0581] Among them, 10V means that the volume (mL) of the reagent used is 10 times the gram weight of the raw material.

[0582] Screening Examples 22 - 27

[0583]

[0584] Similar operations to those in Step 1 of Example 21 were adopted. The feeding amount of Compound 9-1 was 0.1 g, and the reaction was monitored by HPLC. The screening of reaction conditions was as follows:

[0585]

[0586]

[0587] Among them, 10V means that the volume (mL) of the reagent used is 10 times the weight of the raw material in grams. For example, in Screening Example 22, the feeding amount of Compound 9-1 was 0.1 g, and 10V means that the amount of 2M methylamine THF solution used was 1 mL.

[0588] Comparative Example 1

[0589]

[0590] The cost of synthesizing Compound WX011 by the conventional method in the art as shown in the above route is significantly higher (about 5-6 times higher) than that by the route shown in the present application. At the same time, the yield of the route in the present application is increased by 3-4 times compared with the conventional route.

[0591]

[0592] Comparative Example 2

[0593]

[0594] The cost of synthesizing Compound WX021 by the conventional method in the art as shown in the above route is significantly higher (about 3-4 times higher) than that by the route shown in the present application. At the same time, the yield of the synthesis route in the present application is increased by about ten times. Most importantly, the migration of TBS can be effectively avoided, and the target product can be obtained more efficiently and quickly.

[0595]

Claims

1. A method for preparing compound III, which is the following scheme 1 or scheme 2: in, Base is R is H, C1-C4 alkoxy, -OTBS, -OTOM, halogen or -O-MOE; Pg2 n is 0, 1 or 2; R 2 is isopropyl or tert-butyl; R X , R Y and R Z independently H or C 1-6 Alkyl, or, R X and R Z Together with the carbon atom and nitrogen atom between them, they form ring A; ring A is a 4-8 membered nitrogen-containing carbon heterocycle; Option 1: R 1 is H; And the Pg2 is When, it includes the following steps: In the presence of an alkyl tertiary amino base, in a halogenated hydrocarbon solvent, the compound II and the Pg2 protective group precursor B are subjected to a substitution reaction B to obtain the compound III; The Pg2 protecting group precursor B is Where -OR 5 is a leaving group; n is 0, 1 or 2; R 2 is isopropyl or tert-butyl; Option 2: R 1 is H or C1-C4 alkyl-C(═O)-; And the Pg2 is When, it includes the following steps: In solvent A, the compound II and Pg2 protective group precursor A are subjected to substitution reaction A to obtain the compound III; The solvent A is an alcohol solvent, a halogenated hydrocarbon solvent or a nitrogen-containing aromatic compound solvent; The Pg2 protecting group precursor A is Among them, R X , R Y and R Z independently H or C 1-6 Alkyl, or, R X and R Z Together with the carbon atom and nitrogen atom between them, they form ring A; ring A is a 4-8 membered nitrogen-containing carbon heterocycle.

2. The preparation method according to claim 1, characterized in that It meets one or more of the following conditions: (1)R X , R Y and R Z independently H or C 1-4 Alkyl; the C 1-4 The alkyl group is preferably methyl, ethyl or n-butyl; Preferably, R X H or C 1-4 Alkyl; R Y and R Z Independently for C 1-4 alkyl; More preferably, R X is H or methyl; R Y is methyl, ethyl or n-butyl; R Z is methyl, ethyl or n-butyl; (2) When R X and R Z When forming a ring A with the carbon atom and nitrogen atom between them, Pg2 is (3) Pg2 is (4) for Said Preferably 3. The preparation method according to claim 2, characterized in that: It meets one or more of the following conditions: (1) Pg2 is When the Pg2 protecting group precursor A is DMF-DMA; Preferably, the solvent A is an alcohol solvent; (2) Pg2 is When the Pg2 protecting group precursor A is (3) Pg2 is When the Pg2 protecting group precursor A is (4) Pg2 is When the Pg2 protecting group precursor A is (5) Pg2 is When the Pg2 protecting group precursor A is 4. The preparation method according to claim 1, characterized in that: It meets one or more of the following conditions: (1) The Pg2 protecting group precursor B is (2) In the preparation method of the compound III, in the substitution reaction B, the molar ratio of the Pg2 protecting group precursor B to the compound II is 1-15; (3) In the preparation method of the compound III, in the substitution reaction B, the alkyl tertiary amino base is triethylamine or DIPEA; (4) In the preparation method of the compound III, in the substitution reaction B, the molar ratio of the alkyl tertiary amino base to the compound II is 1.5-15; (5) In the preparation method of the compound III, in the substitution reaction B, the halogenated hydrocarbon solvent is DCM; (6) In the preparation method of the compound III, in the substitution reaction B, the concentration of the compound II in the halogenated hydrocarbon solvent is 0.1-1.5 mol / L; (7) In the method for preparing compound III, the temperature of the substitution reaction B is 10-40°C; (8) The substitution reaction B comprises one or more of the following post-treatment steps: liquid separation, washing of the organic phase, concentration, column chromatography separation, and liquid chromatography separation; (9) In the substitution reaction B, The compound II is Accordingly, the compound III is (10) In the preparation method of compound III, in the substitution reaction A, R 1 is H, acetyl or isobutyryl; (11) In the preparation method of the compound III, in the substitution reaction A, the molar ratio of the Pg2 protecting group precursor A to the compound II is 1-10; (12) In the preparation method of compound III, in the substitution reaction A, the alcohol solvent is a C1-C4 alcohol solvent; (13) In the preparation method of the compound III, in the substitution reaction A, the concentration of the compound II in the solvent A is 0.05-0.25 mol / L; (14) In the method for preparing compound III, the temperature of the substitution reaction A is 10-40°C; (15) In the method for preparing compound III, the substitution reaction A includes one or more of the following post-treatment steps: concentration and column chromatography separation; (16) When the R 1 When H is In the substitution reaction A, the compound II is Accordingly, the compound III is (17) When the R 1 When it is acetyl or isobutyryl, In the substitution reaction A, the compound II is Accordingly, the compound III is (18) When the R 1 When H is In the substitution reaction A, the compound II is Accordingly, the compound III is (19) In the preparation method of the compound III, in the substitution reaction A, in the solvent A, the halogenated hydrocarbon solvent is DCM; (20) In the preparation method of compound III, in the substitution reaction A, the nitrogen-containing aromatic compound solvent is pyridine.

5. The preparation method according to claim 4, characterized in that: It meets one or more of the following conditions: (1) When the Pg2 protecting group precursor B is hour, R 5 for n is 0 or 1; R 2 is isopropyl; (2) In the preparation method of the compound III, in the substitution reaction B, the molar ratio of the Pg2 protecting group precursor B to the compound II is 1, 1.1, 1.3, 3.5, 4 or 8; (3) In the method for preparing the compound III, in the substitution reaction B, the molar ratio of the alkyl tertiary amino base to the compound II is 2, 2.6, 4.4, 4.5, 5, 9 or 10; (4) In the preparation method of the compound III, in the substitution reaction B, the concentration of the compound II in the halogenated hydrocarbon solvent is 0.11 mol / L, 0.13 mol / L, 0.15 mol / L, 0.17 mol / L, 0.18 mol / L, 0.23 mol / L, 0.24 mol / L, 0.26 mol / L, 0.57 mol / L or 1.27 mol / L; (5) In the method for preparing compound III, the temperature of the substitution reaction B is 15°C, 20°C, 25°C or 30°C; (6) In the post-treatment step of the substitution reaction B, the liquid separation is to mix the reaction solution with a mixed aqueous solution of sodium carbonate and sodium bicarbonate and then separate the liquids; (7) In the post-treatment step of the substitution reaction B, the washing organic phase is washed with a mixed aqueous solution of sodium bicarbonate and sodium carbonate; (8) In the post-treatment step of the substitution reaction B, the column chromatography separation is eluted with a mixed solvent of petroleum ether, ethyl acetate and triethylamine; (9) In the post-treatment step of the substitution reaction B, the liquid chromatography separation is eluted with acetonitrile as the mobile phase; (10) In the preparation method of the compound III, in the substitution reaction A, the molar ratio of the Pg2 protecting group precursor A to the compound II is 1-6; (11) In the preparation method of compound III, in the substitution reaction A, the alcohol solvent is methanol; (12) In the preparation method of the compound III, in the substitution reaction A, the concentration of the compound II in the solvent A is 0.1-0.2 mol / L; (13) In the method for preparing compound III, the temperature of the substitution reaction A is 15°C, 20°C, 25°C, 30°C or 35°C; (14) In the post-treatment step of the substitution reaction A, the column chromatography separation is eluted with one or more of dichloromethane, methanol, petroleum ether, ethyl acetate and triethylamine.

6. The preparation method according to claim 5, characterized in that: It meets one or more of the following conditions: (1) The Pg2 protecting group precursor B is (2) In the preparation method of the compound III, in the substitution reaction A, the molar ratio of the Pg2 protecting group precursor A to the compound II is 2.5, 3, 4, 4.5 or 6; (3) In the method for preparing the compound III, in the substitution reaction A, the concentration of the compound II in the solvent A is 0.10 mol / L, 0.11 mol / L, 0.12 mol / L, 0.13 mol / L, 0.14 mol / L, 0.16 mol / L or 0.18 mol / L; (4) In the post-treatment step of the substitution reaction A, the column chromatography separation is eluted with a mixed solvent of dichloromethane, methanol and triethylamine, or with a mixed solvent of petroleum ether, ethyl acetate and triethylamine.

7. The preparation method according to claim 1, characterized in that: The following steps are also included: In the presence of an organic base, in a solvent, compound I is subjected to a deprotection reaction to obtain compound II; Wherein, the organic base is ammonia and / or alkylamine; The solvent is an ether solvent and / or an alcohol solvent; R 1 is H; Base is Pg1 is C1-C4 alkyl-C(=O)- or C6-C 14 Aryl-C(═O)-; Pg1 is directly connected to the amino group of Base; R is H, C1-C4 alkoxy, -OTBS, -OTOM, halogen or -O-MOE.

8. The preparation method according to claim 7, characterized in that: It meets one or more of the following conditions: (1) In the deprotection reaction, the C1-C4 alkyl -C(=O)- is acetyl or isobutyryl; (2) In the deprotection reaction, the C6-C 14 The aryl-C(=O)- is benzoyl; (3) In the deprotection reaction, the organic base is ammonia or an alkylamine; the alkylamine is preferably methylamine, ethylamine or triethylamine; (4) In the deprotection reaction, the molar ratio of the organic base to the compound I is 5-80, for example, 10, 12, 17.2, 17.4, 17.6, 19.8, 20, 34, 36, 58 or 59; (5) In the deprotection reaction, the solvent is an ether solvent or an alcohol solvent; the ether solvent is preferably THF; the alcohol solvent is preferably a C1-C4 alcohol solvent, such as methanol; (6) In the deprotection reaction, the concentration of the compound I in the solvent is 0.05-0.35 mol / L, for example, 0.06 mol / L, 0.1 mol / L, 0.11 mol / L, 0.12 mol / L, 0.13 mol / L, 0.25 mol / L or 0.27 mol / L; (7) The temperature of the deprotection reaction is 10-35°C, for example, 15°C, 20°C, 25°C or 30°C; (8) The deprotection reaction comprises one or more of the following post-treatment steps: Concentration, separation, recrystallization, column chromatography separation; Wherein, the separation is preferably carried out by mixing the reaction solution with water and methyl tert-butyl ether and then separating the mixture to obtain an organic phase; Wherein, the recrystallization is preferably carried out with methyl tert-butyl ether and n-hexane, or with ethyl acetate and n-heptane; Wherein, the column chromatography separation is preferably eluted with a mixed solvent of petroleum ether, ethyl acetate and / or triethylamine; (9) In the deprotection reaction, the compound I is Accordingly, the compound II is 9. A method for preparing a compound of formula II, comprising the following steps: In the presence of ammonia and / or alkylamine, in a solvent, the compound I shown is subjected to a deprotection reaction to obtain the compound II shown; The conditions and operations are the same as those described in the deprotection reaction of claim 7 or 8.

10. A compound of formula IV-1 or IV-2:

11. Use of a compound of formula IV-1 or IV-2 as an amino protecting agent; in the use, the compound of formula IV-1 or IV-2 replaces the hydrogen atom in -NH- or -NH2 in a nitrogen-containing organic substance with an amino protecting group; The amino protecting group is The application preferably comprises the following steps: In the presence of an alkyl tertiary amino base, in a halogenated hydrocarbon solvent, the nitrogen-containing organic compound and the compound of formula IV-1 or IV-2 are subjected to substitution reaction B to obtain a product in which the hydrogen atom in -NH- or -NH2 in the nitrogen-containing organic compound is replaced with the amino protecting group; The conditions and operations therein are all as described in the conditions and operations in the substitution reaction B of any one of claims 1 to 5.

12. The use according to claim 11, characterized in that The nitrogen-containing organic matter is an amino nucleoside; The amino nucleoside is preferably Among them, Base is R 1 is H; R is H, C1-C4 alkoxy, -OTBS, -OTOM, halogen or -O-MOE.

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