The present invention relates to novel pyrrole substituted 2-indolinone compounds and physiologically acceptable salts and prodrugs thereof which modulate the activity of protein kinases and therefore are expected to be useful in the prevention and treatment of protein kinase related cellular disorders such as cancer

Pyrrole substituted 2-indolinone protein kinase inhibitors

Fluorinated analogues of the canonical α-L-amino acids have gained widespread attention as building blocks that may endow peptides and proteins with advantageous biophysical, chemical and biological properties. This critical review covers the literature dealing with investigations of peptides and proteins containing fluorinated analogues of the canonical amino acids published over the course of the past decade including the late nineties. It focuses on side-chain fluorinated amino acids, the carbon backbone of which is identical to their natural analogues. Each class of amino acids—aliphatic, aromatic, charged and polar as well as proline—is presented in a separate section. General effects of fluorine on essential properties such as hydrophobicity, acidity/basicity and conformation of the specific side chains and the impact of these altered properties on stability, folding kinetics and activity of peptides and proteins are discussed (245 references).

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Fluorinated amino acids: compatibility with native protein structures and effects on protein–protein interactions

Eosinophils promote tissue injury and contribute to the pathogenesis of allergen-triggered diseases like asthma, but the chemical basis of damage to eosinophil targets is unknown. We now demonstrate that eosinophil activation in vivo results in oxidative damage of proteins through bromination of tyrosine residues, a heretofore unrecognized pathway for covalent modification of biologic targets in human tissues. Mass spectrometric studies demonstrated that 3-bromotyrosine serves as a specific “molecular fingerprint” for proteins modified through the eosinophil peroxidase-H2O2 system in the presence of plasma levels of halides. We applied a localized allergen challenge to model the effects of eosinophils and brominating oxidants in human lung injury. Endobronchial biopsy specimens from allergen-challenged lung segments of asthmatic, but not healthy control, subjects demonstrated significant enrichments in eosinophils and eosinophil peroxidase. Baseline levels of 3-bromotyrosine in bronchoalveolar lavage (BAL) proteins from mildly allergic asthmatic individuals were modestly but not statistically significantly elevated over those in control subjects. After exposure to segmental allergen challenge, lung segments of asthmatics, but not healthy control subjects, exhibited a >10-fold increase in BAL 3-bromotyrosine content, but only two- to threefold increases in 3-chlorotyrosine, a specific oxidation product formed by neutrophil- and monocyte-derived myeloperoxidase. These results identify reactive brominating species produced by eosinophils as a distinct class of oxidants formed in vivo. They also reveal eosinophil peroxidase as a potential therapeutic target for allergen-triggered inflammatory tissue injury in humans.

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Eosinophils generate brominating oxidants in allergen-induced asthma

Publisher Summary This chapter has adapted the nonsense codon suppression method for the incorporation of unnatural amino acids into membrane proteins in a Xenopus oocyte expression system Combining this method with electrophysiologic analysis allows it to probe structure-function relationships in ion channels and receptors in ways not possible with conventional mutagenesis In the absence of atomic-scale, structural data for membrane proteins, these techniques can provide detailed structural information In the nonsense codon suppression method, Xenopus oocytes are coinjected with two RNAs: (1) mRNA transcribed in vitro from a mutated cDNA containing a TAG nonsense (stop) codon at the position of interest and (2) a suppressor tRNA containing the corresponding anticodon, CUA, and chemically acylated at the end with an amino acid During protein synthesis, the aminoacylated suppressor tRNA directs the incorporation of the amino acid into the desired position of the protein Because the amino acid is appended synthetically, it is not limited to the natural, and many unnatural amino acids have been incorporated into various proteins using this method This chapter describes suppressor tRNA design and synthesis, chemical acylation of the suppressor tRNA, relevant organic synthesis methods, and optimization of mRNA and suppressor tRNA for Xenopus oocyte expression

In vivo incorporation of unnatural amino acids into ion channels in Xenopus oocyte expression system.

Detection of specific reaction products is a powerful approach for dissecting out pathways that mediate oxidative damage in vivo. Eosinophil peroxidase (EPO), an abundant protein secreted from activated eosinophils, has been implicated in promoting oxidative tissue injury in conditions such as asthma, allergic inflammatory disorders, cancer, and helminthic infections. This unique heme protein amplifies the oxidizing potential of H2O2 by utilizing plasma levels of Br- as a cosubstrate to form potent brominating agents. Brominated products might thus serve as powerful tools for identifying sites of eosinophil-mediated tissue injury in vivo; however, structural identification and characterization of specific brominated products formed during EPO-catalyzed oxidation have not yet been reported. Here we explore the role of EPO and myeloperoxidase (MPO), a related leukocyte protein, in promoting protein oxidative damage through bromination and demonstrate that protein tyrosine residues serve as endogenous traps ...

3-Bromotyrosine and 3,5-dibromotyrosine are major products of protein oxidation by eosinophil peroxidase: potential markers for eosinophil-dependent tissue injury in vivo.

Aus dem α-Brom-o-nitr0phenylacetat (I) erhalt man mit Phthalimid-kalium das I Substitutionsprodukt (II), das zu (IIIa) bzw. dem Hydrochlorid (IIIb) verseift wird.

Synthesis and microbiological properties of 3-amino-1-hydroxy-2-indolinone and related compounds.

2-Chlorotyrosine and 2-bromotyrosine, as well as the previously reported 2-fluorotyrosine, were synthesized by hydrolysis of the condensation products from the appropriate benzyl bromide and ethyl acetamidomalonate and were compared with the corresponding 3-halotyrosines as growth inhibitors of Escherichia coli 9723, Streptococcus faecalis 8043 and Lactobacillus plantarum 8014. In contrast to the 2- and 3-fluorotyrosines which were equally effective as growth inhibitors, the 2-chloro- and 2-bromotyrosines were much more effective than the 3-chloro- and 3-bromotyrosines in inhibiting the growth of the three microorganisms. For each of the assay organisms, the growth inhibitions of all three 2-halotyrosines were reversed competitively in varying degrees by tyrosine.

David R. Smith

Papers

Synthesis and microbiological properties of 3-amino-1-hydroxy-2-indolinone and related compounds.

Synthesis and microbiological activities of some monohalogenated analogs of tyrosine

Preparation and antimicrobial properties of the D and L forms of 3-amino-3,4-dihydro-1-hydroxycarbostyril

Synthesis and microbiological properties of 3-amino-1-hydroxy-2-indolinone and related compounds

Synthetic and structural studies of some chloro‐substituted methyl 2,1‐benzisoxazole‐3‐carboxylates and related compounds