Development and application of diazirines in biological and synthetic macromolecular systems

Blencowe, Anton; Hayes, Wayne

doi:10.1039/b501989c

Cited by 131 publications

(161 citation statements)

References 378 publications

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“…Subsequent work has attempted to improve on these pioneering studies by synthesizing anesthetic analogs with incorporated diazirine groups (6,7). The diazirine moiety absorbs light at longer wavelengths and has been widely used in photolabeling studies (8) and a number of different photoactivatable anesthetics have been synthesized. A diazirine alcohol derivative, 3-(2-hydroxylethyl)-3-n-pentyldiazirin (azioctanol) has been synthesized and characterized (7) and shown to label the nicotinic acetylcholine receptor from Torpedo (9).…”

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confidence: 99%

Identification of Anesthetic Binding Sites on Human Serum Albumin Using a Novel Etomidate Photolabel

Bright¹,

Adham²,

Lemaire³

et al. 2007

Journal of Biological Chemistry

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We have synthesized a novel analog of the general anesthetic etomidate in which the ethoxy group has been replaced by an azide group, and which can be used as a photolabel to identify etomidate binding sites. This acyl azide analog is a potent general anesthetic in both rats and tadpoles and, as with etomidate, is stereoselective in its actions, with the R(؉) enantiomer being significantly more potent than the S(؊) enantiomer. Its effects on ␣1␤2␥2s GABA A receptors expressed in HEK-293 cells are virtually indistinguishable from the parent compound etomidate, showing stereoselective potentiation of GABA-induced currents, as well as direct mimetic effects at higher concentrations. In addition, a point mutation (␤2 N265M), which is known to attenuate the potentiating actions of etomidate, also blocks the effects of the acyl azide analog. We have investigated the utility of the analog to identify etomidate binding sites by using it to photolabel human serum albumin, a protein that binds ϳ75% of etomidate in human plasma and which is thought to play a major role in its pharmacokinetics. Using HPLC/mass spectrometry we have identified two anesthetic binding sites on HSA. One site is the well-characterized drug binding site I, located in HSA subdomain IIA, and the second site is also an established drug binding site located in subdomain IIIB, which also binds propofol. The acyl azide etomidate may prove to be a useful new photolabel to identify anesthetic binding sites on the GABA A receptor or other putative targets.Although it is now widely accepted that general anesthetics exert their effects by binding directly to their protein targets (1-3), information on the precise molecular locations of these binding sites has been slow in coming. Most information has been derived from in vitro electrophysiological experiments in which putative anesthetic targets, ion channels, or receptors, are genetically modified. What this approach provides is information on the molecular determinants of anesthetic sensitivity, but it is usually impossible to tell whether these determinants represent portions of anesthetic binding sites or regions of the channel or receptor that are responsible for transducing anesthetic binding into changes in channel gating. In principle, the most direct approach would be to determine a high resolution crystal structure of the ion channel or receptor in question in the presence and absence of anesthetics. Unfortunately, because of the difficulties in crystallizing membrane proteins, such experiments are still some way off. An alternative strategy that has recently had some success is to use anesthetics that have been modified so that they contain photoactivatable groups. When these anesthetic analogs are illuminated with a bright light, they are converted into highly reactive intermediates, which then, hopefully, bind irreversibly to their targets.The idea of using photoaffinity labeling to identify anesthetic binding sites was first implemented using the volatile anesthetic halothane (4, 5). Halothane ...

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confidence: 99%

Identification of Anesthetic Binding Sites on Human Serum Albumin Using a Novel Etomidate Photolabel

Bright¹,

Adham²,

Lemaire³

et al. 2007

Journal of Biological Chemistry

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“…[18][19][20][21] Chemical stability of diazirines allows sophisticated derivatization of probes under various synthetic conditions. On labeling, diazirines are rapidly photolyzed at wavelength around 350 nm to produce carbene that is usually more reactive than nitrene.…”

Section: Development Of New Phenyldiazirine Familymentioning

confidence: 99%

Development and Leading-Edge Application of Innovative Photoaffinity Labeling

Hatanaka

2015

Chem. Pharm. Bull.

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“…There are two major types of carbene precursor used in cmPAL, namely, diazirine and diazo compounds, which have occupied a uniquely important place in PAL. In particular, in comparison with the latter, diazirines generate relatively more reactive carbenes 12,13 and have either relatively high thermal and chemical stability 2 or a much shorter lifetime when activated by photoirradiation. The potential of carbene precursors as photoreactive groups, especially 3H-aryldiazirines with an absorption wavelength ranging from 340 to 380 nm, was rst revealed in 1973 by Jeremy Knowles.…”

Section: Introductionmentioning

confidence: 99%

“…The boom in cmPAL, as well as the recent development of various diazirine-compatible reactions 12 and the increasing commercial availability of diazirine derivatives, has inspired equally innovative advances in research into small molecule-protein and macromolecule-protein interactions and ligand-gated ion channels. In the last few decades, a great deal of effort has been dedicated to characterizing many classes of enzymes such as kinases, g-secretases, methyltransferases, metalloproteinases and histone deacetylases (HDACs) by using cmPAL in combination with chemical probes, namely, affinity-based probes (AfBPs), in which cmPAL is used to create covalent interactions between the probe and an enzyme that does not employ a catalytic nucleophilic amino acid within the ligand-binding site.…”

Section: Introductionmentioning

confidence: 99%

Current advances of carbene-mediated photoaffinity labeling in medicinal chemistry

Chen

et al. 2018

RSC Adv.

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Photoaffinity labeling (PAL) in combination with a chemical probe to covalently bind its target upon UV irradiation has demonstrated considerable promise in drug discovery for identifying new drug targets and binding sites. In particular, carbene-mediated photoaffinity labeling (cmPAL) has been widely used in drug target identification owing to its excellent photolabeling efficiency, minimal steric interference and longer excitation wavelength.Specifically, diazirines, which are among the precursors of carbenes and have higher carbene yields and greater chemical stability than diazo compounds, have proved to be valuable photolabile reagents in a diverse range of biological systems. This review highlights current advances of cmPAL in medicinal chemistry, with a focus on structures and applications for identifying small molecule-protein and macromolecule-protein interactions and ligand-gated ion channels, coupled with advances in the discovery of targets and inhibitors using carbene precursor-based biological probes developed in recent decades.

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Development and application of diazirines in biological and synthetic macromolecular systems

Cited by 131 publications

References 378 publications

Identification of Anesthetic Binding Sites on Human Serum Albumin Using a Novel Etomidate Photolabel

Identification of Anesthetic Binding Sites on Human Serum Albumin Using a Novel Etomidate Photolabel

Development and Leading-Edge Application of Innovative Photoaffinity Labeling

Current advances of carbene-mediated photoaffinity labeling in medicinal chemistry

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