US 12,018,306 B2
Method for chemically synthesizing Helicobacter pylori core lipopolysaccharide oligosaccharide antigen carbohydrate chain
Jian Yin, Wuxi (CN); Jing Hu, Wuxi (CN); and Xiaopeng Zou, Wuxi (CN)
Assigned to JIANGNAN UNIVERSITY, Wuxi (CN)
Filed by Jiangnan University, Wuxi (CN)
Filed on Sep. 27, 2023, as Appl. No. 18/373,423.
Application 18/373,423 is a continuation of application No. PCT/CN2023/083341, filed on Mar. 23, 2023.
Claims priority of application No. 202210498276.1 (CN), filed on May 9, 2022.
Prior Publication US 2024/0068001 A1, Feb. 29, 2024
Int. Cl. C12P 19/04 (2006.01)
CPC C12P 19/04 (2013.01) 6 Claims
 
1. A method for chemically synthesizing a Helicobacter pylori lipopolysaccharide core oligosaccharide antigen, wherein an H. pylori undecasaccharide antigen shown in formula 1 is synthesized through nine monosaccharide block compounds shown in formula 2 to formula 9 and formula 24;

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wherein PG2, PG6, PG8, PG9, PG12, PG13, PG14, PG15, PG17, PG18, PG19, PG22, PG23, PG25, PG26, PG28, PG29, PG30, PG34, PG35, PG36, and PG37 are independently hydrogen, benzyl, 2-naphthylmethyl tert-butyldimethylsilyl, tert-butyldiphenylsilyl or triethylsilyl;
PG1 is any of hydrogen, methyl, ethyl, tert-butyl, or benzyl;
PG4 and PG5 form propylidene;
PG7, PG11, PG16, PG21, PG27, PG32, PG33, and R38 are selected from the group consisting of acetyl, chloroacetyl, benzoyl, pivaloyl, acetylpropionyl, 9-pentamethoxycarbonyl, 2-naphthylmethyl, and 2-p-methoxybenzyl;
PG10 is one of acetyl, chloroacetyl, benzoyl, pivaloyl, acetylpropionyl, or 9-pentamethoxycarbonyl;
PG20 is monochloroacetyl; PG24 is benzyl; PG31 is benzyl;
a Linker is —(CH2)n—N—Y1Y2 or —(CH2)n—S—Y1, wherein n=1-10, and Y1 and Y2 are independently hydrogen, benzyl, 2-naphthylmethyl, or benzylmethoxycarbonyl; and
LG is a leaving group, and is selected from any of halogen, trichloroacetimidate, N-phenyltrifluoroacetimidate glycoside, methylthio, ethylthio, phenylthio, p-tolylthio, and dibenzyl phosphate;
the method comprising the following steps:
(a) performing a glycosylation reaction between a saccharide block donor formula 3 and a saccharide block receptor formula 2 to prepare a disaccharide compound 11 by the following synthesis route:

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(b) removing the PG7 protecting group of the disaccharide 11 selectively to prepare disaccharide 12; and performing a glycosylation reaction between the disaccharide 12 and a saccharide block donor formula 4 in the presence of an accelerator to prepare a trisaccharide compound 13 by the following synthesis route:

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(c) removing the PG11 protecting group of the trisaccharide compound 13 selectively to prepare trisaccharide 14; and performing a glycosylation reaction between the trisaccharide 14 and a saccharide block donor formula 5 in the presence of an accelerator to prepare a tetrasaccharide compound 15 by the following synthesis route:

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(d) removing the PG16 protecting group of the tetrasaccharide 15 selectively to prepare tetrasaccharide 16; and performing a glycosylation reaction between the tetrasaccharide 16 and a saccharide block donor formula 6 in the presence of an accelerator to prepare a pentasaccharide compound 17 by the following synthesis route:

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OG Complex Work Unit Chemistry
(e) performing a glycosylation reaction between a saccharide block donor formula 7 and a saccharide block receptor formula 9 under the catalysis of an accelerator to prepare disaccharide 18; and removing PG29 of the disaccharide 18 selectively to prepare disaccharide 19 and further performing a glycosylation reaction between the disaccharide 19 and a saccharide block donor formula 7 under the catalysis of an accelerator to prepare a trisaccharide module 20; and after R38 is removed, performing a reaction of the terminal hydroxyl with trichloroacetonitrile or phenyltrifluoroacetyl chloride under an alkaline catalyst to prepare a trisaccharide donor 21 by the following synthesis route:

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(f) removing the PG21 protecting group of the pentasaccharide compound 17 selectively to prepare pentasaccharide 22; and performing a glycosylation reaction between the pentasaccharide 22 and a trisaccharide donor 21 in the presence of an accelerator to prepare an octasaccharide compound 23 by the following synthesis route:

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(g) performing a glycosylation reaction between a glycosyl donor formula 7 and a receptor formula 24 under the catalysis of an accelerator to prepare disaccharide 25; and removing R38 of the disaccharide 25 selectively, and performing a reaction of the terminal hydroxyl with trichloroacetonitrile or phenyltrifluoroacetyl chloride under an alkaline catalyst to prepare a disaccharide receptor 26 by the following synthesis route:

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(h) removing the PG20 protecting group of the octasaccharide 23 selectively to prepare octasaccharide 27; performing a glycosylation reaction between the octasaccharide 27 and a monosaccharide donor formula 8 under an accelerator to prepare a nonasaccharide compound 28; and removing the protecting group PG32 of the nonasaccharide 28 selectively to prepare a nonasaccharide receptor 29, and performing a glycosylation reaction between the nonasaccharide receptor 29 and a glycosyl donor 26 under an accelerator to prepare undecasaccharide 30 by the following synthesis route:

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and
(i) removing an acyl protecting group of the undecasaccharide 30 under an alkaline conditions and an aromatic protecting of the undecasaccharide group under a palladium on carbon/hydrogen condition to complete deprotection, resulting in a completely deprotected H. pylori lipopolysaccharide core undecasaccharide antigen as shown in formula 1.