The present invention concerns fusion of Fc domains with biologically active peptides and a process for preparing pharmaceutical agents using biologically active peptides. In this invention, pharmacologically active compounds are prepared by a process comprising: a) selecting at least one peptide that modulates the activity of a protein of interest; and b) preparing a pharmacologic agent comprising an Fc domain covalently linked to at least one amino acid of the selected peptide. Linkage to the vehicle increases the half-life of the peptide, which otherwise would be quickly degraded in vivo. The preferred vehicle is an Fc domain. The peptide is preferably selected by phage display, E. coli display, ribosome display, RNA-peptide screening, or chemical-peptide screening.

Modified peptides as therapeutic agents

We report a new quantitative metabolome profiling technique based on differential 12C-/13C-isotope dansylation labeling of metabolites, fast liquid chromatography (LC) separation and electrospray ionization Fourier-transform ion cyclotron resonance mass spectrometry (ESI-FTICR MS) detection. An isotope reagent, 13C-dansyl chloride, can be readily synthesized. This reagent, along with 12C-dansyl chloride, provides a simple and robust means of labeling metabolites containing primary amine, secondary amine, or phenolic hydroxyl group(s). It is shown that dansylation labeling offers 1−3 orders of magnitude ESI signal enhancement over the underivatized counterparts. Dansylation alters the chromatographic behaviors of polar and ionic metabolites normally not retainable on a reversed phase (RP) column to an extent that they can be retained and separated by RPLC with high efficiency. There is no isotopic effect on RPLC separation of the differential isotope labeled metabolites, and 12C-/13C-labeled isoforms of me...

Differential 12C-/13C-Isotope Dansylation Labeling and Fast Liquid Chromatography/Mass Spectrometry for Absolute and Relative Quantification of the Metabolome

Tetrahedron report number 186

Biologically active conjugates are disclosed which are formed by reaction of a thiol moiety of a biologically active molecule with a non-peptidic polymer having an active sulfone moiety. Also disclosed are compounds having the formula R1-X-R2 wherein at least one of R1 and R2 is a biologically active molecule having a reactive thiol moiety which forms a covalent bond with X, a Michael acceptor-activated non-peptidic polymer. Further disclosed are methods of making the conjugates and compounds of the present invention as well as pharmaceutical compositions containing them. In addition, activated polymers suitable for attachment to a variety of molecules and surfaces are disclosed.

Pegylation reagents and compounds formed therewith

The chemistry of acetylenic and allenic sulfones

Diphenylphosphinsaurecyanid und Diphenylthiophosphinsaurecyanid reagieren selektiv mit der Mercaptogruppe—auch bei Anwesenheit von Verbindungen mit primaren Aminogruppen. Die Thioloesterbindung in Thiophosphin-, Thiophosphon- und Thiophosphorsaure-S-estern wird durch nicht solvatisierte Fluoridionen ([R4N]F°3 H2O (R = C4H9) und CsF) unter schonenden Bedingungen in Chloroform, Methylenchlorid oder THF gespalten. (C6H5)2P(O)CN und (C6H5)2P(S)CN sind als Schutzgruppen-Reagentien fur Mercaptane geeignet. Diphenylphosphinsaurechlorid und Diphenylthiophosphinsaurechlorid sind in Konkurrenzversuchen mit Butylamin/Butylmercaptan N-selektiv. Bei Cysteamin und Cysteinmethylester wird aber im Gegensatz zu den entsprechenden S-selektiven Cyaniden keine Selektivitat beobachtet. Diphenylphosphinicacidcyanide and diphenylthiophosphinicacidcyanide react selectively with the mercaptogroup even in presence of primary aminogroups. The thioloesterbond in thiophosphinic-, thiophosphonic- and thiophosphoricacid-S-este...

PHOSPHORORGANISCHE VERBINDUNGEN 97: Die Diphenylphosphinyl- und die Diphenylthiophosphinylgruppe als selektive Schutzgruppe für die Mercaptofunktion. Nucleophile Ablösung der Thioloestergruppe aus Thiophosphin-, Thiophosphon-und Thiophosphorsäure-S-estern durch nicht solvatisierte Fluoridionen

Aryl vinyl sulfones selectively react with thiol groups to acid-stable compounds which easily are cleaved by mild bases to thiol compounds. Ester and amide groups are not attacked under these conditions. Therefore, these compounds are good protecting groups or labels in peptide chemistry. The following aryl vinyl sulfones were investigated: phenyl vinyl sulfone 1, p-carbethoxyphenyl vinyl sulfone 15 and the fluorescent 5-dimethylaminonaphthyl 1-vinyl sulfone 5 and 5-methoxynaphthyl-1 vinyl sulfone 10. The last two compounds are very useful reagents for the quantitative determination of SH-groups in polypeptides such as enzymes. Aryl-vinylsulfone reagieren selektiv mit Thiolgruppen zu Verbindungen, die saurestabil sind, aus denen aber unter milden basischen Bedingungen die Thiolverbindung wieder freigesetzt werden kann. Ester- und Amidbindungen werden nicht angegriffen, so das die Anwendung in der Peptidchemie moglich ist. Untersucht werden die Reaktionen von Phenylvinylsulfon 1 sowie p-Carbethoxy...

Aryl-vinylsulfone—reagentien zum schutz und nachweis von thiolfunktionen

Phosphinic-acid-, phosphonic-acid- and phosphoric-acid-derivatives react in high yields with Lawesson's reagent to form the corresponding thiophosphyl-compounds. Phosphinsaure-, Phosphonsaure- und Phosphorsaure-derivate setzen sich mit Lawesson's Reagenz 1 in guten Ausbeuten zu den entsprechenden Thiophosphylverbindungen um.

Phosphororganische Verbindungen 1001 Thiophosphylverbindungen Durch-Austausch

As a result of competition reactions triphenylvinylphosphonium bromide 1 is SH-selective. But the reactivity is significantly reduced in comparison to the vinyl sulfones. The elimination reaction (b) is also slowed down compared with the cleavage of the corresponding sulfones. In diphenylvinylphosphine oxide 2 the SH-selectivity is preserved. The rate of the reactions (a) and (b) is so extremely slow that 2 has no chance as a SH-protective reagent. Triphenyl-vinylphosphoniumbromid 1 reagiert in Konkurrenzversuchen mit RXH (X=O, S, NH) SH-selektiv bei deutlich reduzierter Reaktivitat im Vergleich zu den Vinylsulfonen. Auch die Eliminierung nach (b) verlauft langsamer als bei analogen Sulfonen. Beim Diphenyl-vinylphosphinoxid 2 ist die SH-Selektivitat noch erhalten. Die Reaktionen (a) und (b) verlaufen jedoch so extrem langsam, das 2 als SH-Schutzgruppe praktisch ausscheidet.

Hans Lindel

Papers

PHOSPHORORGANISCHE VERBINDUNGEN 97: Die Diphenylphosphinyl- und die Diphenylthiophosphinylgruppe als selektive Schutzgruppe für die Mercaptofunktion. Nucleophile Ablösung der Thioloestergruppe aus Thiophosphin-, Thiophosphon-und Thiophosphorsäure-S-estern durch nicht solvatisierte Fluoridionen

Aryl-vinylsulfone—reagentien zum schutz und nachweis von thiolfunktionen

Phosphororganische Verbindungen 1001 Thiophosphylverbindungen Durch-Austausch

Phosphororganische verbindungen 1121 reaktivität und selektivität von vinylphosphoniumsalzen und vinylphosphinoxiden gegenüber nucleophilen

Fluoreszierende und nicht fluoreszierende (Aryl)(vinyl)sulfone — Reagenzien zum Schutz und Nachweis von Thiolfunktionen