3 research outputs found
Allylation and Alkylation of Biologically Relevant Nucleophiles by Diallyl Sulfides
Allyl
sulfides are bioactive phytochemicals found in garlic, onion,
and other members of the genus <i>Allium</i>. Here we showed
that diallyl disulfide and diallyl trisulfide can transfer allyl side
chains to low molecular weight thiols. Diallyl monosulfide is inert
with respect to this allyl transfer reaction. On the other hand, diallyl
sulfone, a known metabolite of diallyl monosulfide, alkylates both
amines and thiols under physiologically relevant conditions via isomerization
to an electrophilic vinyl sulfone
Reactions of 1,3-Diketones with a Dipeptide Isothiazolidin-3-one: Toward Agents That Covalently Capture Oxidized Protein Tyrosine Phosphatase 1B
Protein tyrosine phosphatase 1B (PTP1B)
is a validated therapeutic
target for the treatment of type 2 diabetes; however, the enzyme has
been classified by some as an “undruggable target”.
Here we describe studies directed toward the development of agents
that covalently capture the sulfenyl amide “oxoform”
of PTP1B generated during insulin signaling events. The sulfenyl amide
residue found in oxidized PTP1B presents a unique electrophilic sulfur
center that may be exploited in drug and probe design. Covalent capture
of oxidized PTP1B could permanently disable the intracellular pool
of enzyme involved in regulation of insulin signaling. Here, we employed
a dipeptide model of oxidized PTP1B to investigate the nucleophilic
capture of the sulfenyl amide residue by structurally diverse 1,3-diketones.
All of the 1,3-diketones examined here reacted readily with the electrophilic
sulfur center in the sulfenyl amide residue to generate stable covalent
attachments. Several different types of products were observed, depending
upon the substituents present on the 1,3-diketone. The results provide
a chemical foundation for the development of agents that covalently
capture the oxidized form of PTP1B generated in cells during insulin
signaling events
Covalent Allosteric Inactivation of Protein Tyrosine Phosphatase 1B (PTP1B) by an Inhibitor–Electrophile Conjugate
Protein tyrosine
phosphatase 1B (PTP1B) is a validated drug target,
but it has proven difficult to develop medicinally useful, reversible
inhibitors of this enzyme. Here we explored covalent strategies for
the inactivation of PTP1B using a conjugate composed of an active
site-directed 5-aryl-1,2,5-thiadiazolidin-3-one 1,1-dioxide inhibitor
connected via a short linker to an electrophilic α-bromoacetamide
moiety. Inhibitor–electrophile conjugate <b>5a</b> caused
time-dependent loss of PTP1B activity consistent with a covalent inactivation
mechanism. The inactivation occurred with a second-order rate constant
of (1.7 ± 0.3) × 10<sup>2</sup> M<sup>–1</sup> min<sup>–1</sup>. Mass spectrometric analysis of the inactivated enzyme
indicated that the primary site of modification was C121, a residue
distant from the active site. Previous work provided evidence that
covalent modification of the allosteric residue C121 can cause inactivation
of PTP1B [Hansen, S. K., Cancilla, M. T., Shiau, T. P., Kung, J.,
Chen, T., and Erlanson, D. A. (2005) <i>Biochemistry</i> <i>44</i>, 7704–7712]. Overall, our results are
consistent with an unusual enzyme inactivation process in which noncovalent
binding of the inhibitor–electrophile conjugate to the active
site of PTP1B protects the nucleophilic catalytic C215 residue from
covalent modification, thus allowing inactivation of the enzyme via
selective modification of allosteric residue C121