Total synthesis, structural, and biological evaluation of stylissatin A and related analogs.

The natural product cyclic peptide stylissatin A (1a) was reported to inhibit nitric oxide production in LPS-stimulated murine macrophage RAW 264.7 cells. In the current study, solid-phase total synthesis of stylissatin A was performed by using a safety-catch linker and yielded the peptide with a trans-Phe(7) -Pro(6) linkage, whereas the natural product is the cis rotamer at this position as evidenced by a marked difference in NMR chemical shifts. In order to preclude the possibility of 1b being an epimer of the natural product, we repeated the synthesis using d-allo-Ile in place of l-Ile and a different site for macrocyclization. The resulting product (d-allo-Ile(2) )-stylissatin A (1c) was also found to have the trans-Phe(7) -Pro(6) peptide conformations like rotamer 1b. Applying the second route to the synthesis of stylissatin A itself, we obtained stylissatin A natural rotamer 1a accompanied by rotamer 1b as the major product. Rotamers 1a, 1b, and the epimer 1c were separable by HPLC, and 1a was found to match the natural product in structure and biological activity. Six related analogs 2-7 of stylissatin A were synthesized on Wang resin and characterized by spectral analysis. The natural product (1a), the rotamer (1b), and (d-allo-Ile(2) )-stylissatin A (1c) exhibited significant inhibition of NO(.) . Further investigations were focused on 1b, which also inhibited proliferation of T-cells and inflammatory cytokine IL-2 production. The analogs 2-7 weakly inhibited NO(.) production, but strongly inhibited IL-2 cytokine production compared with synthetic peptide 1b. All analogs inhibited the proliferation of T-cells, with analog 7 having the strongest effect. In the analogs, the Pro(6) residue was replaced by Glu/Ala, and the SAR indicates that the nature of this residue plays a role in the biological function of these peptides. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.

Synthesis, Characterization, and Physicochemical Studies Of Orientation-Controlled Multi-Arm PEG Zn(II)/Mg(II) (Aza)Phthalocyanines.

Tuning the amphiphilicity of (aza)phthalocyanine hydrophobic cores by introducing multiple polyethylene glycol (PEG) moieties with controlled orientations of their (non)peripheral positions is an innovative approach to fabricating water-soluble macrocyclic materials. Although many water-soluble PEGylated macrocycles have been produced in this way, the ability to generate substances with PEG tails oriented outward from the macrocyclic plane in order to obtain non-aggregated, water soluble forms remains a challenge. In this study, we resolved this issue by developing a methods for the synthesis of four new dual directional PEG containing Zn(II)/Mg(II) amphiphiles (ZnPc-PEG, MgPc-PEG, ZnAzaPc-PEG and MgAzaPc-PEG). In addition, the non-aggregating behaviour, and photophysical and photochemical properties of these PEG-complexes were elucidated.

PS-COD and PS-9-BBN: polymer-supported reagents for solution-phase parallel synthesis.

1,5-Cyclooctadiene was deprotonated under LICKOR conditions and reacted with Merrifield resin to afford an immobilized cyclooctadiene in high yield. This polymer is effective as a halogen scavenger, while hydroboration leads to a supported 9-BBN analogue. The latter exhibits similar regioselectivity to 9-BBN in olefin hydroboration. [reaction: see text]

Ionic liquid acceleration of solid-phase suzuki-miyaura cross-coupling reactions.

[reaction: see text] Room-temperature ionic liquids promote various transition metal-catalyzed reactions in the solution phase. Here, for the first time, we show that these effects are translatable to solid-phase reactions. The Suzuki-Miyaura cross-coupling of 4-iodophenol immobilized on polystyrene-Wang resin with various arylboronic acids was significantly accelerated by the ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate ([bmim][BF(4)(-)]).

A method for detecting pyrimidine dimers in the DNA of bacteria irradiated with low doses of ultraviolet light.

Cells of Escherichia coli treated with lysozyme and Brij-58 become permeable to proteins, but do not release their DNA. I incubated permeable cells with an endonuclease that produces single-strand breaks in DNA-containing pyrimidine dimers. The enzyme entered the permeable cells, and, if they had been irradiated with ultraviolet light, caused breaks in their DNA. The frequency of breaks was estimated from the sedimentation pattern of the DNA in alkaline sucrose gradients. The procedure is sensitive enough to detect the dimers produced by a dose of 10 erg/mm(2) at 254 nm, or about 50 dimers per E. coli genome. This method exemplifies and extends the use of permeabilized cells for examining biological processes at the molecular level.

Solid-phase synthesis of peptidomimetic oligomers with a phosphodiester backbone.

An unnatural biopolymer is described in which amino acid side-chains are presented along a negatively charged phosphodiester backbone. For this purpose, a series of phosphoramidite monomers was prepared from chiral 1,2-diols. These were efficiently converted into oligomers using standard coupling conditions on an automated DNA synthesizer.

The effect of lexA and recF mutations on post-replication repair and DNA synthesis in Escherichia coli K-12.

We have examined lexA1 uvrA6 and recF143 uvrBdelta derivatives of Escherichia coli K-12 for post-replication repair and DNA synthesis after UV irradiation. Compared to corresponding lex+ rec+ strains, we found that the lexA and recF cells were defective in (1) converting short DNA segments synthesized after irradiation to DNA of normal size; (2) synthesizing high molecular weight DNA after irradiation; (3) transferring pyrimidine dimers from irradiated DNA into unirradiated daughter strands. Our results support the hypothesis that after UV irradiation the formation of large DNA molecules in excision-deficient cells of E.coli depends directly or indirectly upon joining short DNA segments into longer strands, concomitant with the transfer of DNA from irradiated tamplates into unirradiated daughter strands. This process appears to require the activity of lexA and recF genes.