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Non-radioactive labels such as fluorescent, chemi- or bio-luminiscent labels are being used extensively for detection of biomolecules in nanogram or microgram quantities and are replacing the traditional and accurate but intrinsically hazardous radioactive labels [1]. Recently, Jaouen and others have explored a new method for labeling protein and other biomolecules with transition metal carbonyl compounds such as Arene-Cr(CO)3 [2] and Fischer carbene complexes [3,6], etc. Fischer carbene complexes [4] are especially attractive because of their extraordinarily high reactivity towards pendent amino groups of biomolecules and characteristic strong IR signals [5] at 1900¨C2100 cm1 in the IR absorption where no organic molecule absorbs. However, Fischer carbene complexes are generally hydrophobic and insoluble in aqueous medium. This presents a serious bottleneck for their adaptation to biological applications. Mixtures of solvents such as water/acetonitrile have been used [6] but proteins tend to loose their conformational integrity (hence catalytic function) in such mixed medium [7] and might thus interfere with assay results. This prompted us to design and develop synthesis of water-soluble Fischer carbene complexes that are biocompatible. To impart hydrophilicity to a Fischer carbene core, it appeared to us, one could incorporate a sugar moiety [8], an ionic group like NMeþ 3 or an oligoethylene glycol/polyethylene glycol (OEG/PEG) tether (Chart 1). The present study deals with the last two possibilities. It may be pertinent to mention that incorporation of polyethylene glycol (peg) to the Fischer carbene complex would also constitute a method for the pegylation of a protein [9]. Pegylation is the process incorporating PEG chains to a molecule [10]. Pegylated drugs or proteins exhibit increased stability (more resistant to proteolysis), decreased immunogenicity and increased circulating life [11]. In some cases peg conjugation was also found to confer targeting properties to the disease site such as tumor masses by passive diffusion [12]. Effectiveness of pegylation has been demonstrated by the discovery of pegylated drugs such as PEG-IFN-a2 which, in combination with ribavirin, is considered as the gold standard of therapy for hepatitis C [13]. In the present study, we report synthesis of relatively hydrophilic Fischer carbene complexes several of which feature PEG/OEG groups. |
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