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PH-induced structural changes of surface immobilized poly(l-lysine) by two-dimensional (2D) infrared correlation study

This paper reports the pH-induced structural changes in the surface immobilized poly(l-lysine) (PLL) film. Two-dimensional (2D) correlation analysis was applied to the Fourier transform infrared (FTIR) spectra of the surface-immobilized PLL film to examine the spectral changes induced by the alternations of the protonation state of the amino group in the side chain. Significant spectral changes in the FTIR spectra of the PLL film were observed between pH 7 and 8. The decrease in the protonation state of the amino group in the side chain induced spectral changes in the amino group as well as conformational changes in the alkyl group in the side chain. From pH 1-8, the spectral changes in the amino and alkyl groups in the side chain occurred before those of the amide group in the main chain of the surface immobilized PLL film.

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Organometallic-polypeptide block copolymers: Synthesis and self-assembly of poly(ferrocenyldimethylsilane)-b-poly(epsilon-benzyloxycarbonyl-L-lysine)

A new type of metallopolymer-polypeptide block copolymer poly(ferrocenyldimethylsilane)-b-poly (e-benzyloxycarbonyl-L-lysine) was synthesized by ring-opening polymerization of epsilon-benzyloxycarbonyl-L-lysine N-carboxyanhydride initiated by using a primary amino-terminated poly(ferrocenyldimethylsilane). Studies on the self-organization behavior of this polypeptide block copolymer in both the bulk state and in solution were performed. In the bulk state, a cylindricalin-lamellar structure was observed in a compositionally asymmetric sample. Rod-like micelles with a polyferrocenylsilane core formed in a polypeptideselective solvent alone or in the presence of a common solvent. Significantly, an additional small quantity of the common solvent assisted the formation of longer micelles and micelles with better shape-regularity. This is attributed to a decrease in the number of nucleation events and PFS core reorganization effects.

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Co-delivery of Cu(I) chelator and chemotherapeutics as a new strategy for tumor theranostic

Chelating Cu from tumors has been verified as an effective and promising strategy for cancer therapy through antiangiogenesis. However, systematic removal Cu by injecting with Cu chelators will result unavoidable side effects, since Cu is indispensable to the body. In this work, a micelle targeting to tumors’ newborn vessels based on a polypeptide was developed to co-load DOX and Probe X, which can go through an ?OFF-to-ON? procedure to report the Cu+-capture events in vivo in a real-time way by giving near infrared (NIR) fluorescence and photoacoustic signal. By co-delivering antiangiogenesis and chemotherapeutic reagents, the tumor can be significantly suppressed, meanwhile with a low systematic toxicity. Hopefully, this work can offer new insights in designing sophisticated antitumor strategy.

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Reversible PEGylation and Schiff-base linked imidazole modification of polylysine for high-performance gene delivery

Gene carriers made from polylysine are of interest in relation to gene therapy but suffer from the lack of transfection efficiency due to limited stability and endosomal escape ability. To address this problem, we designed and developed Schiff-base linked imidazole modified polylysine with a reversible-PEGylation catiomer (SL-ImPEG-SS-PLL) for high efficiency gene delivery. The reversible PEGylation was introduced for in vivo circulation, as well as selective PEG detachment to augment the cellular internalization, while introducing Schiff-base linked imidazole residues into polylysine was expected to accelerate the endosomal escape of the DNA payload, as well as facilitate intracellular DNA unpacking and release, thus significantly enhancing gene delivery efficiency. The size alteration of the SL-I15mPEG-SS-PLL/pDNA polyplexes in the presence of 10 mM GSH suggests stimulus-induced PEG detachment under tumor relevant reduction conditions. Acid-base titration assays indicate that imidazole residues confer on polylysine remarkable buffering ability. The agarose gel retardation assay suggests that the Schiff-base linkages provide an increased DNA binding ability to protect DNA against nucleases and timely intracellular DNA unpacking to permit DNA dissociation from polylysine and access to transcriptional machinery. Biological efficacy assessment of this multifunctional carrier, using pEGFP and pGL-3 as reporter genes, indicates comparable to or even higher transfection efficiencies than gold standard PEI and their transfection efficiencies are slightly affected by serum. More importantly, in vivo transfection of pEGFP reveals that GFP expression was found not only in some of the important organs, such as the liver, spleen, kidneys and lungs, but also in the transplanted carcinoma. These experimental results suggest that the reversible PEGylation and Schiff-base linked imidazole modification make SL-ImPEG-SS-PLL a great potential candidate for an effective and biocompatible gene delivery system. This journal is

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SYNTHESIS OF AMINO ACID, N-CARBOXYANHYDRIDES

A process for producing N-carboxyanhydrides is disclosed. The process produces N-carboxyanhydrides as a product and HCl as a by-product. The HCl by-product is purged from the reaction mixture by passing a purge gas through the reaction mixture as a carbonylation reagent is reacting with an amino acid or a salt thereof. The N-carboxyanhydrides produced by this process have a relatively lower chloride impurity content, relatively higher yields can be achieved, and the N-carboxyanhydrides can be produced on a larger scale.

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pH-sensitive polymeric micelles assembled by stereocomplexation between PLLA-b-PLys and PDLA-b-mPEG for drug delivery

pH-responsive stereocomplexed micelles based on poly(l-lactic acid)-b-polylysine/poly(d-lactic acid)-b-methoxy poly(ethylene glycol) (PLLA-b-PLys/PDLA-b-mPEG) were fabricated by stereocomplexation between enantiomeric PLA segments. The morphology, critical micelle concentration, formation of stereocomplexation and pH sensitivity of the stereocomplexed micelles were investigated by TEM, DLS, fluorescence spectra, DSC, XRD, and the acid-base titration method. Interestingly, it was observed that compared to PLLA-b-PLys micelles, the stereocomplexed micelles showed lower critical micelle concentration. Anticancer drug doxorubicin (DOX) was encapsulated in the stereocomplexed micelles and then they were incubated with Hela cells to study the in vitro anti-tumor effect. The results showed that the DOX-loaded stereocomplexed micelles exhibited a slower drug release behavior and a weaker efficacy of intracellular proliferation inhibition than PLLA-b-PLys micelles. Stereocomplexation would provide a favorable platform to construct stable micelles for cancer therapy.

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SYNTHESIS AND STRUCTURAL STUDY BY X-RAY DIFFRACTION OF LYOTROPIC BLOCK LIPOPEPTIDIC POLYMERS WITH POLYLYSINE AND POLY(GLUTAMIC ACID) PEPTIDIC CHAINS

Amphipatic lipopeptides with polylysine or poly(glutamic acid) peptidic chains have been synthesized.Their structural study, in the dry state and in water solution, by X-ray diffraction, has shown the existence of two types of mesophases: lamellar and hexagonal.The influence of the nature and the degree of polymerization of the peptidic chains and of the water concentration on the type and the structural parameters of the mesophases has been established.

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Useful synthetic method of polypeptides with well-defined structure by polymerization of activated urethane derivatives of alpha-amino acids

A facile polymerization strategy for synthesis of polypeptides with well-defined structure using urethane derivatives of alpha-amino acids in the presence of amine is reported. N-(phenoxycarbonyl) alpha-amino acid was polymerized at 60C in N,N-dimethyl acetamide (DMAc). The obtained polypeptides were adjusted by varying the initial ratio of urethane derivatives to amine, and the molecular weight distribution remained narrow (below 1.35). MALDI-TOF-MS revealed that the obtained polypeptides had amine-incorporated initiating end and amino group at propagating end without any side reactions, terminating chain growth. The block copolypeptides were established by polymerization of urethane derivatives in the presence of amine, followed by subsequent addition of other urethane derivatives. Copyright

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Discovery of (S)-Benzyl (4-(2,5-dioxooxazolidin-4-yl)butyl)carbamate

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Fluorinated PEG-Polypeptide Polyplex Micelles Have Good Serum-Resistance and Low Cytotoxicity for Gene Delivery

A novel PEGylation polypeptide, poly(ethylene glycol)-b-poly(l-lysine)-b-poly(l-cysteine) (PEG-PLL-PCys) triblock copolymer is synthesized via the sequential ring-opening polymerization of amino acid N-carboxyanhydrides initiated by methoxypolyethylene glycol amine (mPEG-NH2, Mw is 2 kDa). Subsequently, the obtained polypeptide is partially conjugated with fluorocarbon chains via disulfide exchange reaction. PLL segment can condense plasmid DNA through an electrostatic force to form a complex core, PEG segment surrounding the complex like a corona can prevent the complex from precipitation and reduce the adsorption of serum, while PCys segment with fluorocarbon can enhance the cellular uptake and the stability of the formed polyplex micelles in physiological conditions. Experiment results exhibit that the fluorinated polypeptides have low cytotoxicity and good gene transfection efficiency even in the presence of 50% fetal bovine serum. (Figure presented.).

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High potency and broad-spectrum antimicrobial peptides synthesized via ring-opening polymerization of alpha-Aminoacid-N-carboxyanhydrides

Antimicrobial peptides (AMPs), particularly those effective against methicillin-resistant Staphylococcus aureus (S. aureus) and antibiotic-resistant Pseudomonas aeruginosa (P. aeruginosa), are important alternatives to antibiotics. Typical peptide synthesis methods involving solid-phase sequential synthesis are slow and costly, which are obstacles to their more widespread application. In this paper, we synthesize peptides via ring-opening polymerization of alpha-amino acid N-carboxyanhydrides (NCA) using a transition metal initiator. This method offers high potential for inexpensive synthesis of substantial quantities of AMPs. Lysine (K) was chosen as the hydrophilic amino acid and alanine (A), phenylalanine (F), and leucine (L) as the hydrophobic amino acids. We synthesized five series of AMPs (i.e., P(KA), P(KL), P(KF), P(KAL), and P(KFL)), varied the hydrophobic amino acid content from 0 to 100%, and determined minimal inhibitory concentrations (MICs) against clinically important Gramnegative and Gram-positive bacteria and fungi (i.e., Escherichia coli (E. coli), P. aeruginosa, Serratia marcescens (S. marcescens), and Candida albicans (C. albicans). We found that P(K10F 7.5L7.5) and P(K10F15) show the broadest activity against all five pathogens and have the lowest MICs against these pathogens. For P(K10F7.5L7.5), the MICs against E. coli, P. aeruginosa, S. marcescens, S. aureus, and C. albicans are 31 mug/mL, 31 mug/mL, 250 mug/mL, 31 mug/mL, and 62.5 mug/mL, while for P(K10F15) the respective MICs are 31 mug/mL, 31 mug/mL, 250 mug/mL, 31 mug/mL, and 125 mug/mL. These are lower than the MICs of many naturally occurring AMPs. The membrane depolarization and SEM assays confirm that the mechanism of microbe killing by P(K10F 7.5L7.5) copeptide includes membrane disruption, which is likely to inhibit rapid induction of AMP-resistance in pathogens.

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