Ortho- and para-(difluoromethyl)aryl-β-d-glucosides: A new class of enzyme-activated irreversible inhibitors of β-glucosidases

Halazy, S.; Berges, V.; Ehrhard, A.; Danzin, Charles

doi:10.1016/0045-2068(90)90007-r

Cited by 88 publications

(54 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The second example is a difluorotolyl glycoside (Fig. 7B) whose aglycone, upon enzymatic release, again liberates hydrogen fluoride, but this time yielding a reactive quinone methide (Halazy et al, 1990). Interestingly, a natural product that functions as a time-dependent inactivator of glycosidases, salicortin, has been isolated from plants of the family Salicaceae (Clausen et al, 1990).…”

Section: S C Withers and R Aebersoldmentioning

confidence: 99%

Approaches to labeling and identification of active site residues in glycosidases

1995

View full text Add to dashboard Cite

Glycosidases play a key role in a number of biological processes and, as such, are of considerable clinical and biotechnological importance. Knowledge of the identities of catalytically important active site residues is essential for understanding the catalytic mechanism, for enzyme classification, and for targeted bioengineering of glycosidases with altered characteristics. Here we review and discuss traditional strategies and novel approaches based on tandem mass spectrometry for the identification of the key active site residues in glycosidases.

show abstract

Section: S C Withers and R Aebersoldmentioning

confidence: 99%

Approaches to labeling and identification of active site residues in glycosidases

1995

View full text Add to dashboard Cite

show abstract

“…Less common are the more selective mechanism-based inhibitors whose efficacy depends upon binding and subsequent enzymatic action to generate a reactive species. These include the conduritol epoxides (Legler, 1968(Legler, , 1970, the quinone methidegenerating glycosides (Halazy et al, 1990;Briggs et al, 1992), and the glycosylmethyl triazenes (Marshall et al, 1980;Sinnott and Smith, 1976). Interestingly, two naturally occurring inhibitors of this class have now been described: the hydroxymethylconduritol epoxide, cyclophellitol (Atsumi et al, 1990;Withers and Umezawa, 1991), isolated from Phellinus sp.…”

mentioning

confidence: 99%

Mechanism-based Inhibition of Yeast α-Glucosidase and Human Pancreatic α-Amylase by a New Class of Inhibitors

Braun

Brayer

Withers

1995

Journal of Biological Chemistry

106

View full text Add to dashboard Cite

A burst of release of one equivalent of trinitrophenolate observed upon inactivation of human pancreatic ␣-amylase proves the required 1:1 stoichiometry. These are the first mechanism-based inhibitors of this class to be described, and the first mechanism-based inhibitors of any sort for the medically important ␣-amylase. In addition to having potential as therapeutics, compounds of this class should prove useful in subsequent structural and mechanistic studies of these enzymes.Specific inhibitors of glycosidases have proved valuable in a number of applications ranging from mechanistic studies (Legler, 1990;Sinnott, 1990) through their use to study protein glycosylation (Elbein et al., 1984), to possible therapeutic uses such as the control of blood glucose levels via control of the degradation of dietary disaccharides and starch (Truscheit et al., 1981) or control of viral infectivity through interference with normal glycosylation of viral coat proteins (Elbein, 1984;Prasad et al., 1987). A number of naturally occurring reversible glycosidase inhibitors are known such as nojirimycin, castanospermine, swainsonine, and acarbose (Legler, 1990), and these have been subjected to intensive study including the synthesis and testing of a number of analogues. Another class of inhibitors that has been less well studied is that of the covalent, irreversible type, typically affinity labels. These are generally synthetic analogues of sugars containing reactive groups such as epoxides, isothiocyanates and ␣-halocarbonyls as reviewed recently (Legler, 1990;Withers and Aebersold, 1995). Less common are the more selective mechanism-based inhibitors whose efficacy depends upon binding and subsequent enzymatic action to generate a reactive species. These include the conduritol epoxides (Legler, 1968(Legler, , 1970, the quinone methidegenerating glycosides (Halazy et al., 1990;Briggs et al., 1992), and the glycosylmethyl triazenes (Marshall et al., 1980;Sinnott and Smith, 1976). Interestingly, two naturally occurring inhibitors of this class have now been described: the hydroxymethylconduritol epoxide, cyclophellitol (Atsumi et al., 1990;Withers and Umezawa, 1991), isolated from Phellinus sp.; and the putative quinone methide-generating glycoside salicortin, isolated from Salix (Clausen et al., 1990).An additional, relatively recently described class of mechanism-based inhibitor that has proved successful is that of the 2-deoxy-2-fluoro (Withers and Aebersold, 1995;Withers et al., 1987Withers et al., , 1988Withers et al., , 1990. These function as excellent inactivators of retaining glycosidases (glycosidases that hydrolyze the glycosidic linkage with net retention of anomeric configuration) by formation of a stable glycosyl-enzyme intermediate which turns over to product only very slowly. As shown in Scheme 1 for an ␣-glucosidase, the normal mechanism of action of this class of enzyme involves the formation and hydrolysis of a glycosyl-enzyme intermediate with general acid/base catalytic assistance via transition states with subs...

show abstract

“…The azide-equipped skeleton of the suicide substrate 2 was prepared by an established procedure 10,11,17 as shown in Scheme 2. The methyl ester of sialic acid 8 was coupled with 6 in a one-pot reaction 26 via a glycosyl chloride intermediate to yield the α-sialoside 9.…”

Section: Preparation and Biochemical Evaluation Of 4-azide-2-difluoromentioning

confidence: 99%

“…11,22,23 Each enzyme was preincubated with 2, and an aliquot of the solution was added to a solution of 2--(4-methylumbelliferyl)-N-acetylneuraminic acid (4-MU-NANA) to determine the residual activity in 5-min intervals. As shown in Table 1, the mM range of the obtained K I values suggests that a high concentration of 2 is required to inactivate the neuraminidases because of the poor interaction between the enzymes and the aglycone moiety.…”

Section: Preparation and Biochemical Evaluation Of 4-azide-2-difluoromentioning

confidence: 99%

“…The function-specific activation by a neuraminidase (sialidase) and the irreversible labeling ability of the 2-difluoromethylphenyl sialoside have potential as novel drug and diagnostic tools. This 2-difluoromethylphenyl-type glycoside was first reported in 1990 by Danzin et al as a glucosidase inhibitor, 11 and it has been applied to various glycosidases, such as galactosidases, 12 N-acetyl glucosaminidase 13 ; other hydrolases, such as phosphatases, 14 sulfatase, 15 and proteases 16 ; and neuraminidases. 10,[17][18][19] Recently, we reported that another 2-difluoromethylphenyl-type sialoside (1b) (Figure 2a) inhibits the sialidase activity of a Trypanosoma cruzi trans-sialidase (TcTS) and that a host cell infection by T. cruzi is also inhibited by treatment with compound 1b.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Aglycone-focused randomization of 2-difluoromethylphenyl-type sialoside suicide substrates for neuraminidases

Kai

Hinou

Nishimura

2012

Bioorganic & Medicinal Chemistry

View full text Add to dashboard Cite

A selective and potent inhibitor of neuraminidases, a hydrolase that is responsible for processing sialylated glycoconjugates, is a promising drug candidate for various infective diseases. The current study demonstrates that the use of an aglycone-focused library of 2-difluoromethylphenyl -sialosides is an effective technique to find potent and selective mechanism-based labeling reagents for neuraminidases. The focused library was constructed from a 4-azide-2-difluoromethylphenyl sialoside (2) and an alkyne-terminated compound library by a click reaction. The focused library showed different inhibition patterns for two neuraminidases, Vibrio cholerae neuraminidase (VCNA) and human neuraminidase 2 (hNeu2), and the most potent inhibitors for each neuraminidase were selected. A kinetic analysis of the selected inhibitors demonstrated that the modification of the aglycone moiety improved the K I value with little change in the t 1/2 value of the enzyme activity relative to the basic skeleton (2).

show abstract

Ortho- and para-(difluoromethyl)aryl-β-d-glucosides: A new class of enzyme-activated irreversible inhibitors of β-glucosidases

Cited by 88 publications

References 22 publications

Approaches to labeling and identification of active site residues in glycosidases

Approaches to labeling and identification of active site residues in glycosidases

Mechanism-based Inhibition of Yeast α-Glucosidase and Human Pancreatic α-Amylase by a New Class of Inhibitors

Aglycone-focused randomization of 2-difluoromethylphenyl-type sialoside suicide substrates for neuraminidases

Contact Info

Product

Resources

About