59322-44-0

Usage

Type of compound

Synthetic chemical compound

Use

Substrate in laboratory assays to detect and quantify neuraminidase enzyme activity

Modified form

Sialic acid derivative N-acetylneuraminic acid

Modification

Addition of a 4-methylumbelliferyl group at the 2' position

Function

Produces a fluorescent signal when cleaved by neuraminidase

Applications

Enzymatic activity assays, high-throughput screening of neuraminidase inhibitors, influenza research, and drug development targeting neuraminidase.

InChI:InChI=1/C21H25NO11/c1-9-5-16(27)31-15-6-11(3-4-12(9)15)32-21(20(29)30)7-13(25)17(22-10(2)24)19(33-21)18(28)14(26)8-23/h3-6,13-14,17-19,23,25-26,28H,7-8H2,1-2H3,(H,22,24)(H,29,30)/t13-,14+,17+,18?,19+,21+/m0/s1

Upstream product

• 59361-08-9 methyl (4-methylcoumarin-7-yl-5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosid)onate 
• 1434265-18-5 methyl 2,4,7,8,9--penta--O--acetyl-5--tert-butyloxycarbonylamino--3,5--dideoxy--D--glycero--α,β--D-galaco-2--nonulsopyranosonate 
• 73159-08-7 methyl 3,5--dideoxy--D--glycero--α,β--D--galaco--2--nonulsopyranosonate amino glycoside 
• 6931-68-6 methyl 5-acetamido-2,4,7,8,9-penta-O-acetyl-3,5-dideoxy-D-glycero-D-galacto-2-nonulopyranosonate 
• 132883-18-2 methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-D-glycero-β-D-galacto-non-2-ulopyranosyl chloride 
• 107020-85-9 (4S,5R,6R)-4-Acetoxy-5-acetylamino-6-((1S,2R)-1,2,3-triacetoxy-propyl)-tetrahydro-pyran-2-carboxylic acid methyl ester 
• 90-33-5 7-hydroxy-4-methyl-chromen-2-one 
• 118977-25-6 methyl (phenyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-2-thio-D-galacto-2-nonulopyranoside)onate 
• 6770-41-8 Methyl (5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-β-D-glycero-D-galacto-2-nonulopyranosyl chloride)onate 
• 5980-33-6 sodium 4-methylumbelliferonate 
• 67670-69-3 N-acetyl-4,7,8,9-tetra-O-acetyl-2-chloro-2-deoxyneuraminic acid methyl ester 

Downstream Products

• 6813-81-6 N-acetyl neuraminic acid 
• 90-33-5 7-hydroxy-4-methyl-chromen-2-one 
• 489-46-3 sialic acid 
• 126569-16-2 benzyl (4-methylcoumarin-7-yl 5-acetamido-7,8-di-O-acetyl-4,9-di-O-butyldimethylsilyl-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosid)onate 
• 126569-11-7 benzyl (4-methylcoumarin-7-yl 5-acetamido-7-O-acetyl-4-O-butyldimethylsilyl-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosid)onate 
• 126569-15-1 benzyl (4-methylcoumarin-7-yl 5-acetamido-4,9-di-O-butyldimethylsilyl-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosid)onate 
• 126569-13-9 (2S,4S,5S,6R)-6-((1R,2R)-1-Acetoxy-2,3-dihydroxy-propyl)-5-acetylamino-4-(tert-butyl-dimethyl-silanyloxy)-2-(4-methyl-2-oxo-2H-chromen-7-yloxy)-tetrahydro-pyran-2-carboxylic acid benzyl ester 
• 126569-17-3 benzyl (4-methylcoumarin-7-yl 5-acetamido-7,8-di-O-acetyl-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosid)onate 
• 126660-08-0 (2S,4S,5S,6R)-5-Acetylamino-4-(tert-butyl-dimethyl-silanyloxy)-6-[(S)-((R)-2,2-dimethyl-[1,3]dioxolan-4-yl)-hydroxy-methyl]-2-(4-methyl-2-oxo-2H-chromen-7-yloxy)-tetrahydro-pyran-2-carboxylic acid benzyl ester 
• 126569-08-2 benzyl (4-methylcoumarin-7-yl 5-acetamido-4-O-acetyl-8,9-O-isopropylidene-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosid)onate 
• 126569-07-1 benzyl (4-methylcoumarin-7-yl 5-acetamido-8,9-O-isopropyledene-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosid)onate 
• 126569-12-8 benzyl (4-methylcoumarin-7-yl 5-acetamido-7-O-acetyl-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosid)onate 
• 126569-05-9 benzyl (4-methylcoumarin-7-yl-5-acetamido-9-O-acetyl-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosid)onate 
• 126569-09-3 benzyl (4-methylcoumarin-7-yl 5-acetamido-4-O-acetyl-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosid)onate 

Relevant academic research and scientific papers

Facile anomer-oriented syntheses of 4-methylumbelliferyl sialic acid glycosides

As part of a program to find new sialidases and determine their enzymatic specificity and catalytic activity, a library of 4-methylumbelliferyl sialic acid glycosides derivatised at the C-5 position were prepared fromN-acetylneuraminic acid. Both α- and β-4-methylumbelliferyl sialic acid glycosides were prepared in high yields and stereoselectivity. α-Anomers were accessedviareagent control by utilising additive CH3CN and TBAI, whereas the β-anomers were synthesised through a diastereoselective addition reaction of iodine and the aglycone to the corresponding glycal followed by reduction of the resulting 3-iodo compounds. Both anomer-oriented synthetic pathways allow for gram-scale stereoselective syntheses of the desired C-5 modified neuraminic acid derivatives for use as tools to quantify the enzymatic activity and substrate specificity of known sialidases, and potential detection and investigation of novel sialidases.

Fluorogenic sialic acid glycosides for quantification of sialidase activity upon unnatural substrates

Herein we report the synthesis of N-acetyl neuraminic acid derivatives as 4-methylumbelliferyl glycosides and their use in fluorometrically quantifying human and bacterial sialidase activity and substrate specificities. We found that sialidases in the human promyelocytic leukemic cell line HL60 were able to cleave sialic acid substrates with fluorinated C-5 modifications, in some cases to a greater degree than the natural N-acetyl functionality. Human sialidases isoforms were also able to cleave unnatural substrates with bulky and hydrophobic C-5 modifications. In contrast, we found that a bacterial sialidase isolated from Clostridium perfringens to be less tolerant of sialic acid derivatization at this position, with virtually no cleavage of these glycosides observed. From our results, we conclude that human sialidase activity is a significant factor in sialic acid metabolic glycoengineering efforts utilizing unnatural sialic acid derivatives. Our fluorogenic probes have enabled further understanding of the activities and substrate specificities of human sialidases in a cellular context.

PHASE-TRANSFER-CATALYZED SYNTHESIS OF ARYL α-KETOSIDES OF N-ACETYLNEURAMINIC ACID. A 2-METHYLFLUORAN-6-YL GLYCOSIDE OF N-ACETYLNEURAMINIC ACID, 2-METHYL-6-(5-ACETAMIDO-3,5-DIDEOXY-α-D-glycero-D-galacto-NONULOPYRANOSYLONIC ACID)XANTHENE-9-SPIRO-1'-ISOBENZOFURAN-3'-ONE, A NEW SUBSTRATE..

Glycosidation of N-acetylneuraminic acid by phase-transfer catalysis in chloroform-aqueous alkali gave several known and some new aryl α-ketosides in a short reaction time and in good yields.The 4-methylumbelliferyl α-ketoside, the standard substrate for neuraminidase, was prepared in a yield of up to 70percent.New Neu5Ac ketosides were prepared with fluorescein and the fluorescein analog, 2-methyl-6-hydroxyfluoran (2-methyl-6-hydroxyxanthene-9-spiro-1'-isobenzofuran-3'-one) as aglycons, the latter being synthesized from 2-(2-hydroxy-5-methylbenzoyl) benzoic acid and 3-fluorophenol.The α configuration was ascertained by 400-MHz 1H-NMR spectroscopy and by cleavage of the ketosides with neuraminidases from Vibrio cholerae and Clostridium perfringens.The enzymic hydrolysis of the 2-methylfluoran-6-yl ketoside gave Km values of 82 μM (V. cholerae) and 96 μM (C. perfringens).

Reinvestigation of the synthesis of 4-methylcoumarin-7-yl 5-acetamido-3,5-dideoxy-alpha-D-glycero-D-galacto-2-nonulopyranosidonic acid, a fluorogenic substrate for neuraminidase.

A crystalline tetrabutylammonium salt of 7-hydroxy-4-methylcoumarin was prepared and shown to contain two coumarin residues for each ammonium group. Condensation of this salt with the glycosyl chloride of methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-beta-D-glycero-D-galacto-2-nonulopyranosonate in dry acetonitrile at room temperature gave the corresponding alpha-glycoside in higher yield and purity than previously reported methods. Removal of the acetyl and methyl ester blocking-groups gave the free glycoside, which was shown to have the alpha configuration by n.m.r. spectroscopy. In contrast, the reaction of the free coumarin derivative with the chloro sugar in refluxing, dry toluene in the presence of cadmium carbonate as acid acceptor gave none of the above glycoside, but gave the corresponding glycal in good yield.