Synthesis of biocompatible amino acid-modified poly(acrylic acid) derivatives for intracellular gene delivery

Cationic polymers provide versatile traits that fit for nonviral gene delivery applications. However, cationic polymers could exhibit significant cytotoxicity. Poly(acrylic acid) by conjugating side chains with either primary amine or with different number of alpha mine of alanine (denoted as PAla polymers) was chemically modified and the impact of amine groups of poly(acrylic acid) regarding biocompatibility and intracellular gene delivery efficiency was investigated. Design of PAla polymers proved that the incorporation of alanine in cationic polymers may significantly decrease cytotoxicity of the resulting polymers while maintaining the efficiency of cellular uptake and the intracellular gene delivery of the DNA/cationic polymer complex.     

Synthesis and characterization of four‐arm star‐shaped polymers and copolymers

Four‐arm star‐shaped polymers and copolymers were obtained by transition metal‐catalyzed atom‐transfer radical polymerization (ATRP). The polymers were characterized by FTIR and ¹H‐NMR spectroscopy. Gel permeation chromatography results indicated the formation of polystyrene and polystyrene‐block‐poly(methyl methacrylate) (PS‐b‐PMMA) arms with controlled molecular weights. In dilute solution, the linear polymers had higher inherent viscosities than star‐shaped ones. Thermogravimetric analysis showed a similar degradation mechanism for linear and star‐shaped polymers. Differential scanning calorimetry indicated the successful formation of diblock star‐shaped copolymers. Copyright © 2006 Society of Chemical Industry     

Star polymers with random number of temperature sensitive arms and crosslinked poly(EGDMA)-core and their application to drug delivery

A simple methodology is described for the preparation of temperature sensitive star polymers with random number of arms and crosslinked core. In the first step, well defined, monodisperse poly(N-isopropylacrylamide) PNIPAAm polymers were prepared by reversible addition–fragmentation chain transfer polymerization (RAFT-polymerization) by using 4-cyanopentanoic acid dithiobenzoate as chain transfer agent (CTA). In the second step, the PNIPAAm polymers were used as macro-CTA’s for copolymerization with ethylene glycol dimethacrylate (EGDMA) by the RAFT-technique. Adjusting the macro-CTA to EGDMA ratio, nanometric star polymers with small or big core could be prepared as revealed by gel permeation chromatography (GPC), dynamic light scattering (DLS) and viscometry. The same methodology was applied for the preparation of star polymers with poly(NIPAAm)-b-poly(hexyl acrylate) arms. The so prepared temperature sensitive multiarm materials are a stable alternative to polymeric micelles as nano-carriers for drug-delivery applications.     

Thiol Michael addition reaction: a facile tool for introducing peptides into polymer‐based gene delivery systems

Polymers, as widely used non‐viral gene carriers, suffer from high cytotoxicity and relatively low transfection efficiency. Such crucial drawbacks of polymers could be solved by incorporating short and bioactive peptides. The resulting synthetic polymer–peptide conjugates can not only maintain their own special characteristics, but also gain novel characteristics far beyond those of their parent polymer and peptide components to overcome barriers to gene delivery. There are various chemoselective reactions applied in the synthesis of polymer–peptide conjugates, such as Heck, Sonogashira and Suzuki coupling, Diels–Alder cycloaddition, click chemistry, Staudinger ligation, reductive alkylation and oxime/hydrazone chemistry. Among them, thiol–ene click reactions, including thiol–ene radical and thiol Michael addition reactions, are common methods for preparing peptide–polymer conjugates. In this review, we focus on thiol Michael addition reactions, elaborate on their mechanisms and highlight their applications in the synthesis of polymer–peptide conjugates for gene delivery. © 2017 Society of Chemical Industry     

通过RAFT聚合合成星型聚合物

采用先臂法合成(tBA/HDDA)星型聚合物。由丙烯酸正丁酯(tBA)的可逆加成-断裂链转移自由基聚合(RAFT),得到线型PtBA大分子链转移剂;PtBA与双官能团的偶联剂1,6-己二醇二丙烯酸酯(HDDA)反应得到星型聚合物。研究了偶联剂HDDA与线型PtBA大分子链转移剂的摩尔比对合成星型聚合物的影响。采用GPC法测定了线型大分子链转移剂PtBA及星型聚合物的分子量和分子量分布。结果表明,HDDA与PtBA的比例越高,星型聚合物的产率越大;超过一定值,则产生凝胶。GPC结果表示,所得星型聚合物的分子量大,分子量分布窄(PD I<1.19)。     

星形聚合物的研究与应用进展

回顾了结构性能独特且应用颇为广泛的非线性星形聚合物研究现状;介绍了星形聚合物的特点、应用,以及通过活性聚合制备星形聚合物的不同合成方法,比较了各种合成方法的优缺点;综述了星形聚合物用作研究聚合物的支化模型、简化树形聚合物复杂的支化结构、获得聚合物支化信息的研究进展。     

Star poly(2‐ethyl‐2‐oxazoline)s—synthesis and thermosensitivity

A series of star‐shaped poly(2‐ethyl‐2‐oxazoline)s was prepared by cationic polymerization. The polymerization was initiated by dipentaerythrityl hexakis(4‐nitrobenzene sulfonate) and a tosylated hyperbranched polymer of glycidol. The polymerization proceeded in a controlled manner. The star structure of the products was determined by nuclear magnetic resonance. The molar mass distributions that were measured by gel permeation chromatography with multiangle laser light scattering were narrow, and the experimental values of the molar masses were close to those predicted. The very compact structure of the polymers obtained (compared with the linear counterparts) confirmed the star formation. The star poly(2‐ethyl‐2‐oxazoline)s show a phase transition temperature in the range 62–75 °C. Comparison of this phase transition temperature with that of the linear poly(2‐ethyl‐2‐oxazoline)s with the same molar masses indicates the influence of molar mass and topological structure of the macromolecule on temperature behavior. The prepared copolymers are spherical, which might be useful for the controlled transport and release of active compounds. Copyright © 2011 Society of Chemical Industry     

Gene Delivery by Liposomes

In gene therapy, genetic materials, such as plasmid DNA, antisense oligonucleotides (asODNs), and small interfering RNA (siRNA), can be used to treat or prevent disease. This includes replacing a mutated gene, inactivating a mutated gene, or introducing a new gene. Although gene therapy is a promising treatment option for a number of diseases (including inherited disorders, some types of cancer, and certain viral infections), successful gene therapy is hampered by the lack of effective delivery systems (viral and nonviral). There are several nonviral gene carriers, such as lipids, polymers, and peptides, that can be used for this purpose. Liposomal delivery of nucleic acids, such as plasmid DNA, asODNs, and siRNAs, represents a very promising nanocarrier system that is relatively safe and effective in delivering genes to targeted locations in the body. Lipoid-based delivery systems have also been shown to be stable in serum and plasma, have improved biodistribution, prolonged circulation half-life, and enhanced target tissue selectivity. The most common lipids used in liposomes are cationic lipids, which facilitate binding between their positively charged head group(s) to negatively charged nucleic acids. This review is focused on the most common methods of lipoid-based nanocarriers of nucleic acids for gene therapy in vivo.     

Synthesis and evaluation of a star amphiphilic block copolymer from poly(ε‐caprolactone) and poly(ethylene oxide) as load and release carriers for guest molecules

The use of polymeric materials as the carrier in the controlled release of guest molecules has become an important research area in the polymeric materials science, because of their advantages of the safety, efficacy and patient convenience. One of them, star amphiphilic polymer can self‐assemble into supermolecular structure (polymer micelles) by the balance of hydrophilic and hydrophobic interaction. In this study, star amphiphilic copolymer consisting of hydrophobic and biodegradable poly(ε‐caprolactone) (PCL) and hydrophilic poly(ethylene oxide) (PEO) blocks were synthesized by two‐step ring‐opening polymerization. The resultant polymer was characterizated by FTIR, ¹H‐NMR, and DSC to determine its chemical structure. The morpholoy of the polymer micelles was analyzed by TEM. Using star‐PCL‐b‐PEO as carriers and congo red as model guest molecules, the encapsulation and release properties were investigated by UV–visable analysis. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

In vitro degradation behavior of biodegradable 4-star micelles

Drug delivery vectors for sustained release include a variety of polymeric constituents, both natural and synthetic. Among synthetic polymers several linear block copolymer systems have been explored for use as drug delivery vectors. Release of the pharmaceutical agent is affected by the degradation characteristics and/or by the swelling of the polymer. The goal of this study is to evaluate the degradation behavior of branched polyethylene oxide polylactide polyether ester as a drug delivery vector. Three samples of a star polyethylene oxide/polylactide copolymer with differing polylactide chain lengths were evaluated by characterizing the thermal properties of the neat polymer and in vitro degradation behavior.The thermal and morphological properties were examined by DSC, TGA and XRD. The in vitro polymeric micelle samples were observed over time by UV-vis, TEM and fluorescence. The four star PEO-PLA polymers have exceptional amphiphilic characteristics, which enable their use for a variety of applications. The polymers are thermally stable at biological conditions. In addition, the star polymers have shorter degradation times as compared to previously reported linear PLA and PEG-PLA copolymers, suggesting use as a short-term drug release agent. The four star PEO/PLA copolymer may be an excellent candidate for drug delivery applications.     

Bioreducible poly(amido amine) copolymers derived from histamine and agmatine for highly efficient gene delivery

Histamine (HIS) can facilitate the endosomal escape of polyplexes via the ‘proton sponge effect’ because of its imidazole groups. Agmatine (AGM) can improve the transmembrane process of polyplexes as a result of its guanidinium groups. Therefore, HIS and AGM were used as amino monomers to react with cystamine bisacrylamide (CBA) through Michael addition. The synthesized peptide‐mimicking poly(CBA‐HIS/AGM)s showed high transfection efficiency and low cytotoxicity, indicating their great potential as gene carriers. The results also demonstrated that the effects of HIS and AGM on the properties of poly(CBA‐HIS/AGM)s were different: HIS could increase their buffering capacities and bioreducibility, but AGM could facilitate their plasmid DNA packaging and condensing abilities. In addition, the results of transfection mechanism studies indicated that poly(CBA‐HIS/AGM) polyplexes entered into cells mainly via clathrin‐dependent endocytosis and they could efficiently escape the endosome, indicating endosomal escape was not the limiting step for gene delivery based on these polymers. © 2018 Society of Chemical Industry     

Z-RAFT star polymerization of styrene: Comprehensive characterization using size-exclusion chromatography

Reversible addition-fragmentation chain transfer (RAFT) polymerizations of styrene in bulk at 80 °C using tri-, tetra-, and hexafunctional trithiocarbonates, in which the active RAFT groups are linked to the core via the stabilizing Z-group, were studied in detail. These Z-RAFT star polymerizations of styrene showed excellent molecular weight control up to very high monomer conversions and star sizes of more than 200 kDa. The application of high pressure up to 2600 bar was found to significantly increase the relative amount of living star polymer. Not even at very high monomer conversions and for large star molecules, a shielding effect of growing arms hampering the RAFT process could be identified. Absolute molecular weights of star polymers using a conventionally calibrated SEC setup were determined with high precision by using a mixture of linear and star-shaped RAFT agents. When using phenylethyl as the leaving R-group, well-defined star polymers that perfectly match the theoretical predictions were formed. However, when using benzyl as the leaving group, a pronounced impact of monomer conversion on the star polymer topology was observed and pure star polymers with the expected number of arms could not be obtained.     

pH responsive polymers with amino acids in the side chains and their potential applications

Design and synthesis of pH responsive polymeric materials has become an important subject in academia as well as in industrial field in recent years due to their applications in diverse field including controlled drug delivery, biomedical applications, membrane science, sensors and actuators, oil recovery, colloid stabilization, etc. Efforts have been made to incorporate stimuli‐responsive biomolecules in synthetic polymers to develop pH responsive “smart” non‐biological hybrid macromolecules with high water solubility, enhanced biocompatibility, bio‐mimetic structure and properties. This review is focused on the recent advances in side‐chain amino acid‐based pH responsive polymers synthesis and potential application aspects of these macromolecular architectures in drug and gene delivery, and other fields. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 41084.     

Novel polyisobutylene stars. XXIII. Thermal,mechanical, and processing characteristics of poly(phenylene ether)/polydivinylbenzene(polyisobutylene‐b‐polystyrene)37 blends

The objective of these investigations was to increase the use temperature of novel star‐block polymers consisting of a crosslinked polydivinylbenzene (PDVB) core from which radiate multiple poly(isobutylene‐b‐polystyrene) (PIB‐b‐PSt) arms, abbreviated by PDVB(PIB‐b‐PSt)_n. We achieved this objective by blending star‐blocks with poly(phenylene oxide) (PPO) that is miscible with PSt. Thus, various PPO/PDVB(PIB‐b‐PSt)_n blends were prepared, and their thermal, mechanical, and processing properties were investigated. The hard‐phase glass‐transition temperature of the blends could be controlled by the amount (wt %) of PPO. The blends displayed superior retention of tensile strengths at high temperatures as compared to star blocks. The melt viscosities of blends with low weight percentages of PPO were lower than those of star blocks. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 86: 2866–2872, 2002     

Synthesis,characterization and surface properties of star‐shaped polymeric surfactants with polyhedral oligomeric silsesquioxane core

Star‐shaped amphiphilic polymeric surfactants comprising a hydrophobic polyhedral oligomeric silsesquioxane (POSS) core and hydrophilic poly(ethylene glycol) (PEG) arms with various chain lengths are successfully synthesized using copper(I)‐catalysed azide–alkyne cycloaddition (CuAAC) click reaction. Their chemical structures and molecular characteristics are clearly confirmed using Fourier transform infrared and ¹H NMR spectroscopies and gel permeation chromatography, and no homopolymer is found after CuAAC click reaction. Aqueous solutions of these star‐shaped polymers have been investigated using atomic force and transmission electron microscopies and dynamic light scattering studies and it is found that they can self‐assemble into micelles. The sizes of the micelles can be adjusted by the length of the PEG arms, where longer chains not only lead to increased micelle sizes, but also reduce the contact angle values. Moreover, the melting points and root mean square roughness of the obtained star‐shaped polymers are slightly increased on increasing the chain length of the PEG arms. © 2017 Society of Chemical Industry     

Co‐delivery of methotrexate and doxorubicin via nanocarriers of star‐like poly(DMAEMA‐block‐HEMA‐block‐AAc) terpolymers

Combined therapy is a promising strategy for clinical cancer treatment with synergistic effects. The purpose of the work reported was to evaluate a smart nanocarrier for co‐delivery of doxorubicin (DOX) and methotrexate (MTX). Since star‐like nanocarriers can load a high dose of drugs with various properties, we developed star polymer nanomicelles based on poly[(2,2‐dimethylaminoethyl methacrylate)‐block‐(2‐hydroxyethyl methacrylate)‐block‐(acrylic acid)] having potential for multi‐drug delivery. The nanomicelles demonstrated high encapsulation efficiency, i.e. 97.1% for DOX and 79.5% for MTX. To this end, the star‐like terpolymers were synthesized via atom transfer radical polymerization with pentaerythritol as an initiator. The micellar properties and dual stimuli‐responsive behaviour of the terpolymers were investigated using transmission electron microscopy, field emission scanning electron microscopy and dynamic light scattering measurements, concluding that this co‐therapy offers a promising approach for cancer treatment. © 2019 Society of Chemical Industry     

Polymeric siRNA delivery vectors: knocking down cancers with polymeric‐based gene delivery systems

One of the most promising routes for cancer therapy that has evolved over the previous decade is the use of small‐interfering RNA (siRNA) as a means of switching off genes that are responsible for tumour development. However, while siRNA and gene/antisense therapies provide alternatives to conventional chemotherapies, significant hurdles related to the delivery and efficacy of treatment must still be overcome before this technology can be used as an effective treatment for cancer and other diseases. This review highlights the issues associated with siRNA therapy in vivo, and describes the various approaches that are being explored using polymers as delivery vectors. In particular, the review focuses on targeted delivery as a means of improving efficacy of the gene therapy. © 2014 Society of Chemical Industry     

Synthesis of multi‐arm star polystyrene with hyperbranched polyester initiators by atom transfer radical polymerization

Multi‐arm star polystyrenes with hyperbranched polyester (HP3) core were prepared by atom transfer radical polymerization (ATRP). The structures of the polymers were investigated with FTIR and ¹H NMR. GPC results showed that the resultant polymers had relatively broad polydispersity indices that arouse from the macromolecular initiator (HP3‐Br). The thermal properties were studied using differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). DSC analysis indicated that polystyrene star polymers had only the glass transition temperatures (T_g), which changes with the weight ratio of multi‐functional macroinitiator‐to‐monomer. In addition, these star polymers could form the spherical micelles in the selected solvent (THF/n‐hexane). © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 99: 728–733, 2006     

Phase separation phenomena in aqueous solutions of amphiphilic poly(ethylene oxide) star polymers

Three- and four-armed star polymers with poly(ethylene oxide) arms capped with hydrophobic end-groups were synthesized from nonylphenoxypoly(ethylene glycol)s and well defined tri- and tetraisocyanates. The latter were hydrosilylation products of m-isopropenyl-,-dimethylbenzyl isocyanate (m-TMI). In aqueous solution the arm ends associate, and above a critical star concentration the mixture phase separates into a dilute phase and a condensed gel phase. Their respective polymer concentrations remain constant as long as the two phases coexist, their volume fractions being proportional to the total polymer concentration. Brookfield viscosity measurements confirm the formation of the gel phase which resembles an amphiphilic hydrogel. It exhibits a high affinity for hydrophobic compounds.     

Incorporation of Polycaprolactone to Cyclodextrin‐Based Nanocarrier for Potent Gene Delivery

Stability of polyplex and safety are key factors to achieve stable gene transfection and high transfection efficiency. In this report, a star‐like amphiphilic biocompatible cyclodextrin‐poly(ε‐caprolactone)‐poly(2‐(dimethylamino) ethyl methacrylate), β‐CD‐g‐(PCL‐b‐PDMAEMA) _x copolymer, consisting of biocompatible cyclodextrin core, biodegradable and stable poly(ε‐caprolactone) PCL segments, cationic and hydrophilic PDMAEMA blocks, is synthesized to achieve high efficiency of gene transfection with enhanced stability, due to the micelle formation by hydrophobic PCL segments. In comparison with polyethylenimine (PEI‐25k), a golden standard for nonviral vector gene delivery, this copolymer shows higher encapsulated plasmid desoxyribose nucleic acid (pDNA) ability and the persistence of transgene expression. More interestingly, this gene delivery platform by β‐CD‐g‐(PCL‐b‐PDMAEMA) _x shows lower toxicity but better gene transfection efficiency at low N/P ratios, indicating high potential in gene therapy applications.