MMP2-Targeting and Redox-Responsive PEGylated Chlorin e6 Nanoparticles for Cancer Near-Infrared Imaging and Photodynamic Therapy.
Hou Wenxiu,Xia Fangfang,Alves Carla S,Qian Xiaoqing,Yang Yuming,Cui Daxiang
ACS applied materials & interfaces
A unique matrix metalloproteinase 2-targeted photosensitizer delivery platform was developed in this study for tumor-targeting imaging and photodynamic therapy. The model photosensitizer therapeutic agent chlorin e6 (Ce6) was first covalently conjugated with matrix metalloproteinase 2-cleavable polypeptide and then modified with polyethylene glycol via a redox-responsive cleavable disulfide linker. The resultant matrix metalloproteinase 2-cleavable polypeptide modified PEGylated Ce6 (PEG-SS-Ce6-MMP2) nanoparticles, which formed via self-assembly, were observed to be monodisperse and significantly stable in aqueous solution. In addition, owing to their cellular redox-responsiveness at the cleavable disulfide linker, the PEG-SS-Ce6-MMP2 nanoparticles were able to release Ce6 rapidly. Despite displaying enhanced intracellular internalization, the synthesized PEG-SS-Ce6-MMP2 nanoparticles did not compromise their phototoxic effects toward A549 cancer cells when compared with free Ce6 and PEGylated Ce6 nanoparticles. In vivo experiments further revealed that, in contrast with the free Ce6 or with the PEGylated Ce6 nanoparticles, the PEG-SS-Ce6-MMP2 nanoparticles showed a remarkable increase in tumor-targeting ability and a significantly improved photodynamic therapeutic efficiency in A549 tumor-bearing mice. These results suggest that the PEG-SS-Ce6-MMP2 nanoparticles hold great potential for tumor-targeting imaging and photodynamic therapy.
Silicon Quantum Dot Nanoparticles with Antifouling Coatings for Immunostaining on Live Cancer Cells.
Tu Chang-Ching,Chen Kuang-Po,Yang Tsu-An,Chou Min-Yuan,Lin Lih Y,Li Yaw-Kuen
ACS applied materials & interfaces
Fluorescent silicon quantum dots (SiQDs) have shown a great potential as antiphotobleaching, nontoxic and biodegradable labels for various in vitro and in vivo applications. However, fabricating SiQDs with high water-solubility and high photoluminescence quantum yield (PLQY) remains a challenge. Furthermore, for targeted imaging, their surface chemistry has to be capable of conjugating to antibodies, as well as sufficiently antifouling. Herein, antibody-conjugated SiQD nanoparticles (SiQD-NPs) with antifouling coatings composed of bovine serum albumin (BSA) and polyethylene glycol (PEG) are demonstrated for immunostaining on live cancer cells. The monodisperse SiQD-NPs of diameter about 130 nm are synthesized by a novel top-down method, including electrochemical etching, photochemical hydrosilylation, high energy ball milling, and "selective-etching" in HNO3 and HF. Subsequently, the BSA and PEG are covalently grafted on to the SiQD-NP surface through presynthesized chemical linkers, resulting in a stable, hydrophilic, and antifouling organic capping layer with isothiocyanates as the terminal functional groups for facile conjugation to the antibodies. The in vitro cell viability assay reveals that the BSA-coated SiQD-NPs had exceptional biocompatibility, with minimal cytotoxicity at concentration up to 1600 μg mL(-1). Under 365 nm excitation, the SiQD-NP colloid emits bright reddish photoluminescence with PLQY = 45-55% in organic solvent and 5-10% in aqueous buffer. Finally, through confocal fluorescent imaging and flow cytometry analysis, the anti-HER2 conjugated SiQD-NPs show obvious specific binding to the HER2-overexpressing SKOV3 cells and negligible nonspecific binding to the HER2-nonexpressing CHO cells. Under similar experimental conditions, the immunofluorescence results obtained with the SiQD-NPs are comparable to those using conventional fluorescein isothiocyanate (FITC).
Synthesis and activity of a folate targeted monodisperse PEG camptothecin conjugate.
Henne Walter A,Kularatne Sumith A,Hakenjos John,Carron Joshua D,Henne Kristene L
Bioorganic & medicinal chemistry letters
A folate targeted camptothecin small molecule drug conjugate (SMDC) was synthesized using a monodisperse PEG spacer linked to folate via a releasable disulfide carbonate linker. Cell cytotoxicity in human KB cells exhibited an IC50 of 6nM. Importantly, activity of the prodrug was blocked by excess folate, demonstrating receptor-mediated celluar uptake of the PEG conjugate.
Supramolecular self-assembly of monoend-functionalized methoxy poly(ethylene glycol) and α-cyclodextrin: from micelles to hydrogel.
Long Youyu,Song Hongmei,He Bin,Lai Yusi,Liu Rong,Long Chunyan,Gu Zhongwei
Journal of biomaterials applications
An effective strategy was developed to fabricate a supramolecular hydrogel with the complexation of α-cyclodextrins (α-CDs) and monoend-functionalized low molecular weight methoxy poly(ethylene glycol) (mPEG, Mn=2000) micelles. Hydrophobic cinnamic acid was immobilized on methoxy poly(ethylene glycol) via L-lysine as linker to prepare amphiphilic mPEG. The monoend-functionalized mPEG self-assembled micelles in aqueous solution. The size and size distribution of the micelles were tested by dynamic laser scattering (DLS). The morphology of the micelles was observed by SEM, TEM and AFM. The critical micelle concentration (CMC) was tested and it was 42.5 mg/L. The monodisperse micelles had core-shell structure and the mean diameter was around 40 nanometers. α-cyclodextrins were added in the suspension of micelles to form supramolecular hydrogel with the polypseudorotaxanes complexation. Hydrophilic drug doxorubicin hydrochloride was used as model drug to study the release profile. The results showed that the hydrogel was a promising carrier for drug delivery.
Monodisperse and Polydisperse PEGylation of Peptides and Proteins: A Comparative Study.
Wang Jie,Deng Tao,Liu Yuntai,Chen Kexin,Yang Zhigang,Jiang Zhong-Xing
Although PEGylation is widely used in biomedicine with great success, it suffers from many drawbacks, such as polydispersity, nonbiodegradability, and loss of precursor potency. Recently, the search for polyethylene glycol (PEG) substitutes has attracted considerable attention. Some of the substitutes partially address the drawbacks of PEGs, but sacrifice the "stealth" effect of PEGs and bring in new issues. Herein, we developed monodisperse oligoethylene glycol (M-OEG) polyamides over 5000 Da as biodegradable and monodisperse PEGylation (M-PEGylation) agents, which provided M-PEGylated peptides and proteins with high monodispersity and a biodegradable PEG moiety. Compared to regular PEGylated proteins with a complex "stealth" cloud of PEG, the hydrogen bond interactions between the M-OEG polyamides and proteins provided the M-PEGylated protein with a biodegradable "stealth" cloak. The monodisperse and biodegradable M-PEGylation strategy as well as the peculiar protein-M-OEG polyamide interactions may shed light on many long-lasting issues during the development of PEGylated biologic drugs, such as monodispersity, biodegradability, and tunable conformation.
Peptidic Monodisperse PEG "Comb" as Multifunctional "Add-On" Module for Imaging-Traceable and Thermo-Responsive Theranostics.
Zhu Junfei,Zhang Huaibin,Chen Kexin,Li Yu,Yang Zhigang,Chen Shizhen,Zheng Xing,Zhou Xin,Jiang Zhong-Xing
Advanced healthcare materials
Monodisperse polyethylene glycols-modified (M-PEGylated) biomaterials exhibit high structural accuracy, biocompatibility, and fine-tunable physicochemical properties. To develop "smart" drug delivery systems in a controllable and convenient manner, a peptidic M-PEG "comb" with fluorinated L-lysine side chains and a fluorescent N-terminal is conveniently prepared as a F magnetic resonance imaging ( F MRI) and fluorescence dual-imaging traceable and thermo-responsive "add-on" module for liposomal theranostics in cancer therapy. The peptidic M-PEG "comb" has high biocompatibility, thermo-responsivity with a sharp lower critical solution temperature, an aggregation-induced emission fluorescence, and high F MRI sensitivity. As a highly branched amphiphile, it self-assembles and firmly anchors on the doxorubicin-loaded liposomal nanoparticles, which M-PEGylates the liposomes and facilitates the thermo-responsive drug release and drug tracking with dual-imaging technologies. In a rodent xenograft model of human liver cancer HepG2 cells, the M-PEGylated liposomes exhibit long in vivo half time, low toxicity, high tumor accumulation, "hot spot" F MRI, and therapeutic efficacy. With accurately programmable chemical structure, fine-tunable physicochemical and biological properties to meet the demands of diagnosis, drug delivery, and therapy, the M-PEG "comb" is promising as a versatile "add-on" module for rapid and convenient formulation of various "smart" theranostics.
Transglutaminase-mediated PEGylation of proteins: direct identification of the sites of protein modification by mass spectrometry using a novel monodisperse PEG.
Mero Anna,Spolaore Barbara,Veronese Francesco M,Fontana Angelo
Poly(ethylene glycol) (PEG) has been widely used to prolong the residence time of proteins in blood and to decrease their immunogenicity and antigenicity. A drawback of this polymer lies in its polydispersity that makes difficult the identification of the sites of protein modification. This is a mandatory requirement if a PEGylated protein should be approved as a drug. Here, a fast and reliable method is proposed to characterize proteins conjugated at the level of glutamine (Gln) residues using microbial transglutaminase (TGase). The novelty resides in the use of a monodisperse Boc-PEG-NH(2) for the derivatization that allows the direct identification of the sites of PEGylation by electrospray ionization mass spectrometry (ESI-MS). The procedure has been tested on three model proteins, namely, human granulocyte colony-stimulating factor, human growth hormone, and horse heart apomyoglobin. The Gln residues linked to the polymer chain were easily identified by ESI-MS and tandem MS analyses, demonstrating the advantage of using a monodisperse polymer in combination with mass spectrometry for an easy characterization of conjugated proteins. Interestingly, the PEGylation reaction led to the production only of mono- and bis-derivative products, indicating that the TGase-mediated PEGylation can be extremely selective and thus very useful for the derivatization of protein drugs.
High-performance PEGylated Mn-Zn ferrite nanocrystals as a passive-targeted agent for magnetically induced cancer theranostics.
Xie Jun,Zhang Yu,Yan Caiyun,Song Lina,Wen Song,Zang Fengchao,Chen Gong,Ding Qi,Yan Changzhi,Gu Ning
An effective magnetic nanocrystals (MNCs)-mediated theranostics strategy as a combination of simultaneous diagnostics and heating treatment of tumors by using magnetic resonance imaging (MRI) and alternating current magnetic field (ACMF) is successfully developed. In this strategy, we had firstly synthesized a well-established Mn-Zn ferrite MNCs coated with PEG-phospholipids (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol copolymers, DSPE-PEG2000). The monodisperse PEGylated MNCs with core-shell structure (15 nm) exhibited excellent performance, such as high magnetism of 98 emu g(-1) Fe, relaxivity coefficient (r2) of 338 mm(-1) s(-1), and specific absorption rate (SAR) value of 324 W g(-1) Fe. It was proved that the obtained MNCs with an average diameter of 48.6 nm can drastically minimize the recognition and phagocytosis of macrophages, simultaneously improve their biocompatibility in vitro. These advantages endowed them with efficient passive targeting ability in vivo for prominent tumor MRI and magnetically induced heating when exposed to ACMF, based on enhanced permeability and retention (EPR) effects. To ensure sufficient accumulation of MNCs within tumors for targeted hyperthermia, we described the use of MNCs with a well-tolerated intravenous single dose of 18 mg Fe/kg mouse body weight, achieving repeatedly injection and hyperthermia within a subcutaneous breast cell carcinoma mouse model. With an ACMF of 12 A at 390 kHz, the tumor surface sites could be heated to approximately 43 °C in 30 min based on MNCs-mediated intravenous injections. The long-lasting hyperthermia could effectively induce the apoptosis of tumor cells, inhibit the angiogenesis of tumor vessels, and finally suppress the tumor growth within a certain period of time.
Effect of Chemical Binding of Doxorubicin Hydrochloride to Gold Nanoparticles, Versus Electrostatic Adsorption, on the In Vitro Drug Release and Cytotoxicity to Breast Cancer Cells.
Zayed Gamal M,Kamal Islam,Abdelhafez Wael A,M Alsharif Fahd,Amin Mohamed A,Shaykoon Montaser Sh A,Sarhan Hatem A,Abdelsalam Ahmed M
PURPOSE:The selective delivery of chemotherapeutic agent to the affected area is mainly dependent on the mode of drug loading within the delivery system. This study aims to compare the physical method to the chemical method on the efficiency of loading DOX.HCl to GNPs and the specific release of the loaded drug at certain tissue. METHOD:Bifunctional polyethylene glycol with two different functionalities, the alkanethiol and the carboxyl group terminals, was synthesized. Then, DOX·HCl was covalently linked via hydrazone bond, a pH sensitive bond, to the carboxyl functional group and the produced polymer was used to prepare drug functionalized nanoparticles. Another group of GNPs was coated with carboxyl containing polymer; loading the drug into this system by the means of electrostatic adsorption. Finally, the prepared system were characterized with respect to size, shape and drug release in acetate buffer pH 5 and PBS pH 7.4 Also, the effect of DOX.HCl loaded systems on cell viability was assessed using MCF-7 breast cancer cell line. RESULTS:The prepared nanoparticles were spherical in shape, small in size and monodisperse. The release rate of the chemically bound drug in the acidic pH was higher than the electrostatically adsorbed one. Moreover, both systems show little release at pH 7.4. Finally, cytotoxicity profiles against human breast adenocarcinoma cell line (MCF-7) exhibited greater cytotoxicity of the chemically bound drug over the electrostatically adsorbed one. CONCLUSION:Chemical binding of DOX·HCl to the carboxyl group of PEG coating GNPs selectively delivers high amount of drug to tumour-affected tissue which leads to reducing the unwanted effects of the drug in the non-affected ones.
Monodisperse thermoresponsive microgels of poly(ethylene glycol) analogue-based biopolymers.
Cai Tong,Marquez Manuel,Hu Zhibing
Langmuir : the ACS journal of surfaces and colloids
Monodisperse microgels of P(MEO2MA-co-OEGMA) have been synthesized by using free radical polymerization. Microgels with a variety of particle radii ranging from 82 to 412 nm have been obtained with different surfactant concentrations. The particle size distribution is extremely narrow and even better than that for PNIPAM microgels. Pure MEO2MA microgels have an LCST of about 22 degrees C. The LCSTs corresponding to the molar ratio of OEGMA to MEO2MA at 10 and 20% are 31 and 37 degrees C, respectively. Microgels in water self-assemble into various phases, including a crystalline phase with iridescent colors, which are the result of Bragg diffraction from differently oriented crystalline planes. Considering that PEG is nontoxic and anti-immunogenic as proven by the FDA, thermoresponsive P(MEO2MA-co-OEGMA) microgels may have many exciting biomedical applications.
Preparation of biodegradable PEGylated pH/reduction dual-stimuli responsive nanohydrogels for controlled release of an anti-cancer drug.
Zhou Tingting,Zhao Xubo,Liu Lei,Liu Peng
A facile and efficient method was developed to prepare the monodisperse biodegradable PEGylated pH and reduction dual-stimuli sensitive poly[methacrylic acid-co-poly(ethylene glycol) methyl ether methacrylate-co-N,N-bis(acryloyl)cystamine] (PMPB) nanohydrogels with dried particle size below 200 nm via one-step distillation precipitation polymerization as a drug delivery system (DDS) for the controlled release of a wide-spectrum anti-cancer drug, doxorubicin hydrochloride (DOX). Under normal physiological media, the nanohydrogels possessed high drug encapsulation efficiency (more than 96%) within 48 h and exhibited good stability with a trifle premature drug release. However, rapid DOX release was achieved at lower pH or in the presence of reductive reagent glutathione (GSH) with a cumulative release of more than 85% within 30 h. Furthermore, the nanohydrogels manifested nontoxicity on HepG2 cells at a concentration of 10 μg mL(-1) or lower. Based on the excellent characteristics of the nanohydrogels, such as low toxicity, impressive biodegradability, sharp dual responsiveness, adequate drug loading capacity and a high drug encapsulation efficiency, they were supposed to have potential application in the area of cancer therapy.
Paclitaxel-induced formation of 3D nanocrystal superlattices within injectable protein-based hybrid nanoparticles.
Lee Jeong Yu,Son Ho Yeon,Park Jae Chul,Park Jongnam,Nam Yoon Sung
Chemical communications (Cambridge, England)
Self-assembly of monodisperse superparamagnetic iron oxide nanocrystals into a close-packed, three-dimensional (3D) superlattice is designed within cross-linked protein-based nanoparticles composed of human serum albumin and polyethylene glycol. The prepared nanoparticles are very stable in serum and exhibit a high T2 relaxivity as well as anti-cancer activity, indicating the practical benefits of ordering nanocrystals.
Enhanced Gene Delivery in Bacterial and Mammalian Cells Using PEGylated Calcium Doped Magnetic Nanograin.
Hashemi Ehsan,Mahdavi Hossein,Khezri Jafar,Razi Farideh,Shamsara Mehdi,Farmany Abbas
International journal of nanomedicine
Background:Beyond viral carriers which have been widely used in gene delivery, non-viral carriers can further improve the delivery process. However, the high cytotoxicity and low efficiency impedes the clinical application of non-viral systems. Therefore, in this work, we fabricated polyethylene glycol (PEG) coated, calcium doped magnetic nanograin (PEG/Ca(II)/FeO) as a genome expression enhancer. Methods:Monodisperse magnetic nanograins (MNGs) with tunable size were synthesized by a solvothermal method. The citrate anions on the spherical surface of MNGs capture Ca ions by an ion exchange process, which was followed by surface capping with PEG. The synthesized PEG/Ca(II)/FeO was characterized using Fourier-transform infrared spectroscopy (FTIR), dynamic light scattering (DLS) spectra, vibrating sample magnetometer (VSM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). MTT test was utilized to assess the toxicity of PEG/Ca(II)/FeO. Real time qPCR was applied for quantification of gene expression. Results:DLS spectra and TEM images confirmed a thin layer of PEG on the nanocarrier surface. Shifting the zeta potential in the biological pH window from -23.9 mV (for FeO) to ≈ +11 mV (for PEG/Ca(II)/FeO) confirms the MNGs surface protonation. Cytotoxicity results show that cell viability and proliferation were not hindered in a wide range of nanocarrier concentrations and different incubation times. Conclusion:PEGylated calcium doped magnetic nanograin enhanced PUC19 plasmid expression into and GFP protein expression in HEK-293 T cells compared to control. A polymerase chain reaction of the NeoR test shows that the transformed plasmids are of high quality.
Poly(ethylene glycol) hydroxystearate-based nanosized emulsions: effect of surfactant concentration on their formation and ability to solubilize quercetin.
Dora Cristiana L,Silva Luis F C,Putaux Jean-Luc,Nishiyama Yoshiharu,Pignot-Paintrand Isabelle,Borsali Redouane,Lemos-Senna Elenara
Journal of biomedical nanotechnology
Quercetin is a natural compound that has shown several biological activities. However, it displays poor water solubility and, therefore, low bioavailability. In this study, oil-in-water nanosized emulsions were obtained by the hot solvent diffusion method, using castor oil as oily phase and poly(ethylene glycol) (660)-12-hydroxystearate (PEG 660-stearate) and lecithin as surfactants. The effect of the PEG 660-stearate concentration on the droplet size of the nanosized emulsions and on the ability of these systems to load quercetin was investigated. Dynamic light scattering (DLS), transmission electron microscopy (TEM), cryo-TEM, and small-angle X-ray scattering (SAXS) were used to characterize the systems. We have demonstrated that a critical concentration of PEG 660-stearate (2.5 wt%) was needed to obtain colloidal dispersions displaying microemulsion characteristics. This colloidal dispersion, that was not optically birefringent, was constituted by a monodisperse population of 20 nm-large droplets, and exhibited excellent stability. Besides, this system was able to solubilize five times more quercetin than nanoemulsions prepared using 0.25 wt% PEG 660-stearate. SAXS results suggest that the spherical droplets have a core-shell structure. With regard to the hot solvent diffusion method, both diffusion of the solvent towards the aqueous phase and increase of the temperature above the phase inversion temperature (PIT) of PEG 660-stearate appeared to be required for obtaining clear and isotropic colloidal dispersions.
Enhanced antibiotic activity of ampicillin conjugated to gold nanoparticles on PEGylated rosette nanotubes.
Fan Yiwen,Pauer Alexander C,Gonzales Arthur A,Fenniri Hicham
International journal of nanomedicine
Purpose:This work presents the preparation of a nanocomposite of ampicillin-conjugated gold nanoparticles (AuNPs) and self-assembled rosette nanotubes (RNTs), and evaluates its antibacterial properties against two strains of drug-resistant bacteria ( , methicillin-resistant [MRSA]). Materials and methods:Small, nearly monodisperse AuNPs (1.43±0.5 nm in diameter) nucleated on the surface of polyethylene glycol-functionalized RNTs in a one-pot reaction. Upon conjugation with ampicillin, their diameter increased to 1.86±0.32 nm. The antibacterial activity of the nanocomposite against and MRSA was tested using different concentrations of ampicillin. The cytocompatibility of the nanocomposite was also tested against human dermal fibroblasts. Results:Based on bacterial inhibition studies, the nanocomposite demonstrated enhanced antibiotic activity against both bacterial strains. The minimum inhibitory concentration (MIC) of the nanocomposite against was found to be 0.58 μg/mL, which was 18% lower than ampicillin alone. The nanocomposite also exhibited a 20 hrs MIC of 4 μg/mL against MRSA, approximately 10-20 times lower than previously reported values for ampicillin alone. In addition, at concentrations of 4 μg/mL of ampicillin (70 μg/mL of AuNPs), the nanocomposite showed negligible cytotoxic effects. Conclusion:Our findings offer a new approach for the treatment of drug-resistant bacteria by potentiating inhibitory effects of existing antibiotics, and delivering them using a non-toxic formulation.
Utilizing microfluidics to synthesize polyethylene glycol microbeads for Förster resonance energy transfer based glucose sensing.
Kantak Chaitanya,Zhu Qingdi,Beyer Sebastian,Bansal Tushar,Trau Dieter
Here, we utilize microfluidic droplet technology to generate photopolymerizeable polyethylene glycol (PEG) hydrogel microbeads incorporating a fluorescence-based glucose bioassay. A microfluidic T-junction and multiphase flow of fluorescein isothiocyanate dextran, tetramethyl rhodamine isothiocyanate concanavalin A, and PEG in water were used to generate microdroplets in a continuous stream of hexadecane. The microdroplets were photopolymerized mid-stream with ultraviolet light exposure to form PEG microbeads and were collected at the outlet for further analysis. Devices were prototyped in PDMS and generated highly monodisperse 72 ± 2 μm sized microbeads (measured after transfer into aqueous phase) at a continuous flow rate between 0.04 ml/h-0.06 ml/h. Scanning electron microscopy analysis was conducted to analyze and confirm microbead integrity and surface morphology. Glucose sensing was carried out using a Förster resonance energy transfer (FRET) based assay. A proportional fluorescence intensity increase was measured within a 1-10 mM glucose concentration range. Microfluidically synthesized microbeads encapsulating sensing biomolecules offer a quick and low cost method to generate monodisperse biosensors for a variety of applications including cell cultures systems, tissue engineering, etc.
Organoclay-assisted interfacial polymerization for microfluidic production of monodisperse PEG-microdroplets and in situ encapsulation of E. coli.
Wang Kye Won,Lee Kyoung G,Park Tae Jung,Lee Young-Chul,Yang Ji-Won,Kim Do Hyun,Lee Seok Jae,Park Jung Youn
Biotechnology and bioengineering
We developed a novel one-pot synthetic strategy for preparing monodisperse polyethylene glycol diacrylate (PEGDA) microdroplets via organoclay-assisted interfacial polymerization approach for Escherichia coli encapsulation. Based on the mechanism of spontaneous and rapid polymerization of PEGDA precursor solution with Mg-organoclay, the prepared PEGDA microdroplets have uniform size and fine round shape, with size range of 74-118 µm. The size of microdroplets can be controlled through the changing continuous phase flow rate. Organoclay-assisted polymerization method provides a unique environment to produce non-toxic ways of fabricating microorganism encapsulated microdroplets and to prohibit microdroplets merge during the processes. Furthermore, we successfully carried out to entrap E. coli inside of the PEGDA microdroplets. E. coli expressing a green fluorescent protein shows a good viability inside the PEGDA microdroplets. The in situ microfluidic synthetic method provides a novel approach for the preparation of monodisperse PEGDA microdroplets via a one-pot route.
Colloidal dispersions of monodisperse magnetite nanoparticles modified with poly(ethylene glycol).
Barrera Carola,Herrera Adriana P,Rinaldi Carlos
Journal of colloid and interface science
Monodisperse magnetite nanoparticles modified with poly(ethylene glycol) (PEG) were synthesized using a silane functionalized PEG obtained by reacting 3-aminopropyl triethoxysilane with carboxylic acid-methoxy PEG (mPEG-COOH) using amide reactions. Transmission electron microscopy (TEM), dynamic light scattering (DLS), and zeta potential measurements show the particles are monodisperse (sigma(gv) approximately 0.2) and stable in water for pH of 3-9 and ionic strengths, up to 0.3 M NaCl. Thermogravimetric analysis coupled with TEM and DLS indicates formation of a dense graft layer on the particle surface. An analysis of the interparticle interaction energy indicates that the particles are stabilized by strong steric repulsions between PEG chains on their surface.
Oligo(lactic acid)n-Paclitaxel Prodrugs for Poly(ethylene glycol)-block-poly(lactic acid) Micelles: Loading, Release, and Backbiting Conversion for Anticancer Activity.
Tam Yu Tong,Gao Jieming,Kwon Glen S
Journal of the American Chemical Society
Poly(ethylene glycol)-block-poly(d,l-lactic acid) (PEG-b-PLA) micelles are nanocarriers for poorly water-soluble anticancer agents and have advanced paclitaxel (PTX) to humans due to drug solubilization, biocompatibility, and dose escalation. However, PEG-b-PLA micelles rapidly release PTX, resulting in widespread biodistribution and low tumor exposure. To improve delivery of PTX by PEG-b-PLA micelles, monodisperse oligo(l-lactic acid), o(LA)8 or o(LA)16, has been coupled onto PTX at the 7-OH position, forming ester prodrugs: o(LA)8-PTX and o(LA)16-PTX, respectively. As expected, o(LA)n-PTX was more compatible with PEG-b-PLA micelles than PTX, increasing drug loading from 11 to 54%. While in vitro release of PTX was rapid, resulting in precipitation, o(LA)n-PTX release was more gradual: t1/2 = 14 and 26 h for o(LA)8-PTX and o(LA)16-PTX, respectively. Notably, o(LA)8-PTX and o(LA)16-PTX in PEG-b-PLA micelles resisted backbiting chain end scission, based on reverse-phase HPLC analysis. By contrast, o(LA)8-PTX and o(LA)16-PTX degraded substantially in 1:1 acetonitrile:10 mM PBS, pH 7.4, at 37 °C, generating primarily o(LA)2-PTX. The IC50 value of o(LA)2-PTX was ∼2.3 nM for A549 human lung cancer cells, equipotent with PTX in vitro. After weekly IV injections at 20 mg/kg as PEG-b-PLA micelles, o(LA)8-PTX induced tumor regression in A549 tumor-bearing mice, whereas PTX delayed tumor growth. Surprisingly, o(LA)8-PTX caused less toxicity than PTX in terms of change in body weight. In conclusion, o(LA)n acts as a novel promoiety, undergoing backbiting conversion without a reliance on metabolizing enzymes, and o(LA)n-PTX improves PTX delivery by PEG-b-PLA micelles, providing a strong justification for clinical evaluation.
Oligo(ethylene glycol)-based thermosensitive dendrimers and their tumor accumulation and penetration.
Wu Wei,Driessen Wouter,Jiang Xiqun
Journal of the American Chemical Society
Dendrimers have several featured advantages over other nanomaterials as drug carriers, such as well-defined structure, specific low-nanometer size, and abundant peripheral derivable groups, etc. However, these advantages have not been fully exploited yet to optimize their biological performance, especially tumor penetration, which is a shortcoming of current nanomaterials. Here we show the syntheses of a new class of oligo(ethylene glycol) (OEG)-based thermosensitive dendrimers up to the fourth generation. Each dendrimer shows monodisperse structure. OEG/poly(ethylene glycol) (PEG) moieties with different precise lengths were introduced to the periphery of the fourth-generation dendrimer followed by an antitumor agent, gemcitabine (GEM). The biodistributions of the GEM-conjugated dendrimers were investigated by micro positron emission tomography and multispectral optoacoustic tomography imaging techniques and compared with that of GEM-conjugated poly(amidoamine) (PAMAM). The GEM-conjugated dendrimer with the longest peripheral PEG segments exhibited the most desirable tumor accumulation and penetration and thus had significantly higher antitumor activity than the GEM-conjugated PAMAM.
Toxicity evaluation of monodisperse PEGylated magnetic nanoparticles for nanomedicine.
Patsula Vitalii,Tulinska Jana,Trachtová Štěpánka,Kuricova Miroslava,Liskova Aurelia,Španová Alena,Ciampor Fedor,Vavra Ivo,Rittich Bohuslav,Ursinyova Monika,Dusinska Mária,Ilavska Silvia,Horvathova Mira,Masanova Vlasta,Uhnakova Iveta,Horák Daniel
Innovative nanotechnology aims to develop particles that are small, monodisperse, smart, and do not cause unintentional side effects. Uniform magnetic FeO nanoparticles (12 nm in size) were prepared by thermal decomposition of iron(III) oleate. To make them colloidally stable and dispersible in water and cell culture medium, they were modified with phosphonic acid- (PA) and hydroxamic acid (HA)-terminated poly(ethylene glycol) yielding PA-PEG@FeO and HA-PEG@FeO nanoparticles; conventional γ-FeO particles were prepared as a control. Advanced techniques were used to evaluate the properties and safety of the particles. Completeness of the nanoparticle coating was tested by real-time polymerase chain reaction. Interaction of the particles with primary human peripheral blood cells, cellular uptake, cytotoxicity, and immunotoxicity were also investigated. Amount of internalized iron in peripheral blood mononuclear cells was 72, 38, and 25 pg Fe/cell for HA-PEG@FeO, γ-FeO, and PA-PEG@FeO, respectively. Nanoparticles were localized within the cytoplasm and in the extracellular space. No cytotoxic effect of both PEGylated nanoparticles was observed (0.12-75 μg/cm) after 24 and 72-h incubation. Moreover, no suppressive effect was found on the proliferative activity of T-lymphocytes and T-dependent B-cell response, phagocytic activity of monocytes and granulocytes, and respiratory burst of phagocytes. Similarly, no cytotoxic effect of γ-FeO particles was observed. However, they suppressed the proliferative activity of T-lymphocytes (75 μg/cm, 72 h) and also decreased the phagocytic activity of monocytes (15 μg/cm, 24 h; 3-75 μg/cm, 72 h). We thus show that newly developed particles have great potential especially in cancer diagnostics and therapy.
Synthesis and application of monodisperse oligo(oxyethylene)-grafted polystyrene resins for solid-phase organic synthesis.
Lumpi Daniel,Braunshier Christian,Horkel Ernst,Hametner Christian,Fröhlich Johannes
ACS combinatorial science
In a preliminary investigation by our group, we found that poly(styrene-oxyethylene) graft copolymers (PS-PEG), for example, TentaGel resins, are advantageous for gel-phase (13)C NMR spectroscopy. Because of the solution-like environment provided by the PS-PEG resins, good spectral quality of the attached moiety can be achieved, which is useful for nondestructive on-resin analysis. The general drawbacks of such resins are low loading capacities and the intense signal in the spectra resulting from the PEG linker (>50 units). Here, we describe the characterization of solvent-dependent swelling and reaction kinetics on a new type of resin for solid-phase organic synthesis (SPOS) that allows an accurate monitoring by gel-phase NMR without the above disadvantages. A series of polystyrene-oligo(oxyethylene) graft copolymers containing monodisperse PEG units (n = 2-12) was synthesized. A strong correlation between the linker (PEG) length and the line widths in the (13)C gel-phase spectra was observed, with a grafted PEG chain of 8 units giving similar results in terms of reactivity and gel-phase NMR monitoring to TentaGel resin. Multistep on-resin reaction sequences were performed to prove the applicability of the resins in solid-phase organic synthesis.
Monodisperse carboxyl-functionalized poly(ethylene glycol)-coated magnetic poly(glycidyl methacrylate) microspheres: application to the immunocapture of β-amyloid peptides.
Horák Daniel,Hlídková Helena,Hiraoui Mohamed,Taverna Myriam,Proks Vladimír,Mázl Chánová Eliška,Smadja Claire,Kučerová Zdenka
Identification and evaluation of small changes in β-amyloid peptide (Aβ) levels in cerebrospinal fluid is of crucial importance for early detection of Alzheimer's disease. Microfluidic detection methods enable effective preconcentration of Aβ using magnetic microparticles coated with Aβ antibodies. Poly(glycidyl methacrylate) microspheres are coated with α-amino-ω-methoxy-PEG5000 /α-amino-ω-Boc-NH-PEG5000 Boc groups deprotected and NH2 succinylated to introduce carboxyl groups. Capillary electrophoresis with laser-induced fluorescence detection confirms the efficient capture of Aβ 1-40 peptides on the microspheres with immobilized monoclonal anti-Aβ 6E10. The capture specificity is confirmed by comparing Aβ 1-40 levels on the anti-IgG-immobilized particles used as a control.
Understanding the Influence of a Bifunctional Polyethylene Glycol Derivative in Protein Corona Formation around Iron Oxide Nanoparticles.
Ruiz Amalia,Alpízar Adán,Beola Lilianne,Rubio Carmen,Gavilán Helena,Marciello Marzia,Rodríguez-Ramiro Ildefonso,Ciordia Sergio,Morris Christopher J,Morales María Del Puerto
Materials (Basel, Switzerland)
Superparamagnetic iron oxide nanoparticles are one of the most prominent agents used in theranostic applications, with MRI imaging the main application assessed. The biomolecular interface formed on the surface of a nanoparticle in a biological medium determines its behaviour in vitro and in vivo. In this study, we have compared the formation of the protein corona on highly monodisperse iron oxide nanoparticles with two different coatings, dimercaptosuccinic acid (DMSA), and after conjugation, with a bifunctional polyethylene glycol (PEG)-derived molecule (2000 Da) in the presence of Wistar rat plasma. The protein fingerprints around the nanoparticles were analysed in an extensive proteomic study. The results presented in this work indicate that the composition of the protein corona is very difficult to predict. Proteins from different functional categories-cell components, lipoproteins, complement, coagulation, immunoglobulins, enzymes and transport proteins-were identified in all samples with very small variability. Although both types of nanoparticles have similar amounts of bonded proteins, very slight differences in the composition of the corona might explain the variation observed in the uptake and biotransformation of these nanoparticles in Caco-2 and RAW 264.7 cells. Cytotoxicity was also studied using a standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Controlling nanoparticles' reactivity to the biological environment by deciding on its surface functionalization may suggest new routes in the control of the biodistribution, biodegradation and clearance of multifunctional nanomedicines.
Application of Monodisperse PEGs in Pharmaceutics: Monodisperse Polidocanols.
Yu Zeqiong,Bo Shaowei,Wang Huiyuan,Li Yu,Yang Zhigang,Huang Yongzhuo,Jiang Zhong-Xing
Polydisperse PEGs are ubiquitously used in pharmaceutical industry and biomedical research. However, the monodispersity in PEGs may play a role in the development of safe and effective PEGylated small molecular drugs. Here, to avoid the polydispersity in polidocanol, the active ingredient in a clinically used drug, a macrocyclic sulfate-based strategy for the efficient and scalable synthesis of monodisperse polidocanols, their sulfates, and their methylated derivatives, was developed. TLC and HPLC analysis indicated a complex mixture in regular polidocanol and high purities in monodisperse polidocanols and their derivatives. Assay on HUVEC, L929, and HePG2 cells showed that monodisperse polidocanols have much higher cytotoxicity and safety than that of regular polidocanol. It was found that the monodispersity of PEGs in polidocanols is crucial for achieving the optimal therapeutic results. Therefore, based on this case study, it would be beneficial to optimize PEGylated small molecular drugs with monodisperse PEGs in pharmaceutical research and development.
Protein stabilization by an amphiphilic short monodisperse oligo(ethylene glycol).
Sadhukhan Nabanita,Muraoka Takahiro,Ui Mihoko,Nagatoishi Satoru,Tsumoto Kouhei,Kinbara Kazushi
Chemical communications (Cambridge, England)
A short, monodisperse additive (octa(ethylene glycol) monophenyl ether) functions to suppress aggregation of thermally and chemically denatured lysozyme. Control studies with shorter and non-amphiphilic derivatives revealed that the amphiphilic structure is essential, and octa(ethylene glycol) is nearly the minimum chain length for amphiphilic poly(ethylene glycol)s to stabilize proteins.
Monodisperse oligoethylene glycols modified Camptothecin, 10-Hydroxycamptothecin and SN38 prodrugs.
Deng Tao,Mao Xianglan,Xiao Yan,Yang Zhigang,Zheng Xing,Jiang Zhong-Xing
Bioorganic & medicinal chemistry letters
Camptothecin, which represents a class of natural products with high anticancer activity, suffers low water solubility which hampers its clinic application. To address this issue, monodisperse polyethylene glycols were employed to modify this class of natural products, including Camptothecin, 10-Hydroxycamptothecin, and SN38. Through selective modification with a series of monodisperse polyethylene glycols, 31 Camptothecin derivatives, including 9 ethers and 22 carbonates, were prepared using a macrocyclic sulfate-based strategy with high efficacy. Monodisperse polyethylene glycols modification provided the Camptothecin derivatives with high purity and fine-tunable water solubility. Through the physicochemical and biological assays, a few novel prodrugs with good solubility, cytotoxicity, and valuable drug release profile were identified as promising anticancer drug candidates.
Synthesis, characterization and applications of carboxylated and polyethylene-glycolated bifunctionalized InP/ZnS quantum dots in cellular internalization mediated by cell-penetrating peptides.
Liu Betty R,Winiarz Jeffrey G,Moon Jong-Sik,Lo Shih-Yen,Huang Yue-Wern,Aronstam Robert S,Lee Han-Jung
Colloids and surfaces. B, Biointerfaces
Semiconductor nanoparticles, also known as quantum dots (QDs), are widely used in biomedical imaging studies and pharmaceutical research. Cell-penetrating peptides (CPPs) are a group of small peptides that are able to traverse cell membrane and deliver a variety of cargoes into living cells. CPPs deliver QDs into cells with minimal nonspecific absorption and toxic effect. In this study, water-soluble, monodisperse, carboxyl-functionalized indium phosphide (InP)/zinc sulfide (ZnS) QDs coated with polyethylene glycol lipids (designated QInP) were synthesized for the first time. The physicochemical properties (optical absorption, fluorescence and charging state) and cellular internalization of QInP and CPP/QInP complexes were characterized. CPPs noncovalently interact with QInP in vitro to form stable CPP/QInP complexes, which can then efficiently deliver QInP into human A549 cells. The introduction of 500nM of CPP/QInP complexes and QInP at concentrations of less than 1μM did not reduce cell viability. These results indicate that carboxylated and polyethylene-glycolylated (PEGylated) bifunctionalized QInP are biocompatible nanoparticles with potential for use in biomedical imaging studies and drug delivery applications.
Highly efficient synthesis of monodisperse poly(ethylene glycols) and derivatives through macrocyclization of oligo(ethylene glycols).
Zhang Hua,Li Xuefei,Shi Qiuyan,Li Yu,Xia Guiquan,Chen Long,Yang Zhigang,Jiang Zhong-Xing
Angewandte Chemie (International ed. in English)
A macrocyclic sulfate (MCS)-based approach to monodisperse poly(ethylene glycols) (M-PEGs) and their monofunctionalized derivatives has been developed. Macrocyclization of oligo(ethylene glycols) (OEGs) provides MCS (up to a 62-membered macrocycle) as versatile precursors for a range of monofunctionalized M-PEGs. Through iterative nucleophilic ring-opening reactions of MCS without performing group protection and activation, a series of M-PEGs, including the unprecedented 64-mer (2850 Da), can be readily prepared. Synthetic simplicity coupled with versatility of this new strategy may pave the way for broader applications of M-PEGs.
Monodisperse oligoethylene glycols modified Propofol prodrugs.
Deng Tao,Mao Xianglan,Li Yu,Bo Shaowei,Yang Zhigang,Jiang Zhong-Xing
Bioorganic & medicinal chemistry letters
The low water solubility of Propofol resulted in complicated formulation and adverse effects during its clinical application. To improve its water solubility and maintain its anesthetic effects, Propofol prodrugs with monodisperse oligoethylene glycols as solubility enhancer were designed and synthesized. Monodisperse oligoethylene glycols enable the concise manipulation of water solubility, biocompatibility and anesthetic effects. Through the physicochemical and biological assay, a few water soluble prodrugs of Propofol were identified as promising anesthetic to overcome the drawbacks associated with Propofol.
One-pot synthesis of monodisperse dual-functionalized polyethylene glycols through macrocyclic sulfates.
Lv Xiaoyan,Zheng Xing,Yang Zhigang,Jiang Zhong-Xing
Organic & biomolecular chemistry
Dual-functionalization of monodisperse oligoethylene glycols, especially hetero-functionalization, provides a series of highly valuable intermediates for life and materials sciences. However, the existing methods for the preparation of these compounds suffer excessive protecting and activating group manipulation as well as tedious purification. Here, a one-pot dual-substitution strategy with macrocyclic sulfates of polyethylene glycols as the key intermediates was developed for the convenient and scalable preparation of a series of homo-functionalized and hetero-functionalized oligoethylene glycols in just 1 step. A high synthetic efficacy was achieved by avoiding the protecting and activating group manipulation and the intermediate purification.
Synthesis and characterization of monodisperse poly(ethylene glycol)-conjugated collagen pentapeptides with collagen biosynthesis-stimulating activity.
Kim Myung Sun,Park Eun Ji,Na Dong Hee
Bioorganic & medicinal chemistry letters
Although collagen pentapeptide (Lys-Thr-Thr-Lys-Ser, KTTKS) has received a great deal of attention owing to its collagen biosynthesis-stimulating effects, its enzymatic instability in the skin is an obstacle to effective topical application. PEGylation is a useful approach for improving the chemical and biological stability of peptides. However, the polydispersity of poly(ethylene glycol) (PEG) produces conjugates with different molecular sizes, which may create difficulties in chemical characterization and purity control, and in variability of biological properties. To overcome these difficulties, monodisperse PEG was site-specifically conjugated to the N-terminal amine of KTTKS to produce a single molecular conjugate, enabling more complete chemical characterization and more exact product specifications. PEG-KTTKS conjugates prepared using monodisperse PEG with two different molecular weights, monodisperse PEG220 and PEG572, were characterized by mass spectrometry. These monodisperse PEG-KTTKS conjugates showed no cytotoxicity (1-100 μM) and stimulated collagen biosynthesis in human skin fibroblasts. They also had high stability against proteolytic enzymes in rat skin. This study demonstrates the usefulness of monodisperse PEG for preparing chemically defined conjugates and suggests that monodisperse PEG-KTTKS would be a good candidate for use as a collagen biosynthesis-stimulating agent.
Solid Phase Stepwise Synthesis of Polyethylene Glycols.
Khanal Ashok,Fang Shiyue
Chemistry (Weinheim an der Bergstrasse, Germany)
Polyethylene glycol (PEG) and derivatives with eight and twelve ethylene glycol units were synthesized by stepwise addition of tetraethylene glycol monomers on a polystyrene solid support. The monomer contains a tosyl group at one end and a dimethoxytrityl group at the other. The Wang resin, which contains the 4-benzyloxy benzyl alcohol function, was used as the support. The synthetic cycle consists of deprotonation, Williamson ether formation (coupling), and detritylation. Cleavage of PEGs from solid support was achieved with trifluoroacetic acid. The synthesis including monomer synthesis was entirely chromatography-free. PEG products including those with different functionalities at the two termini were obtained in high yields. The products were analyzed with ESI and MALDI-TOF MS and were found close to monodispersity.
Polyethylene glycol modified magnetic carbon nanotubes as nanosorbents for the determination of methylprednisolone in rat plasma by high performance liquid chromatography.
Yu Panfeng,Ma Hongwei,Shang Yong,Wu Ji,Shen Shun
Journal of chromatography. A
In this paper, polyethylene glycol modified (PEGylated) magnetic carbon nanotubes were developed as solid-phase extraction nanosorbents for the determination of methylprednisolone in rat plasma. The procedure mainly involved two steps including preparation of PEGylated magnetic nanosorbents and bioanalysis. Monodisperse magnetites (Fe3O4) anchored onto multi-walled carbon nanotubes (MWCNTs) were synthesized by a facile solvothermal synthesis method. The obtained MWCNTs-Fe3O4 nanomaterials were further non-covalently functionalized by a surfactant phospholipids-polyethylene glycol (DSPE-PEG). Owing to dispersibility and high enrichment ability, water-soluble PEGylated MWCNTs-Fe3O4 nanomaterials can provide more efficient way for the extraction of methylprednisolone than only MWCNTs-Fe3O4 used. The methylprednisolone could be easily extracted via π-π stacking interactions with PEGylated MWCNTs-Fe3O4. The captured methylprednisolone/nanosorbents were isolated from the matrix by placing a magnet, and desorbed by the elution solvent composed of acetonitrile. Extraction conditions such as amount of nanosorbents added, adsorption time, desorption solvent, and desorption time were investigated and optimized. The method recoveries were obtained from 88.2% to 92.9%. Limits of quantification and limits of detection of 0.01 and 0.005μg/mL were acquired, respectively. The precision ranged from 4.2% to 7.8% for within-day measurement, and for between-day variation was in the range of 5.5-9.0%. Moreover, the analytical performance obtained by PEGylated magnetic MWCNTs was compared with that of magnetic MWCNTs. The results indicated that the approach based on PEGylated magnetic MWCNTs was useful for the analysis of methylprednisolone in the complex plasma.
Fragment-based solid-phase assembly of oligonucleotide conjugates with peptide and polyethylene glycol ligands.
Dirin Mehrdad,Urban Ernst,Noe Christian R,Winkler Johannes
European journal of medicinal chemistry
Ligand conjugation to oligonucleotides is an attractive strategy for enhancing the therapeutic potential of antisense and siRNA agents by inferring properties such as improved cellular uptake or better pharmacokinetic properties. Disulfide linkages enable dissociation of ligands and oligonucleotides in reducing environments found in endosomal compartments after cellular uptake. Solution-phase fragment coupling procedures for producing oligonucleotide conjugates are often tedious, produce moderate yields and reaction byproducts are frequently difficult to remove. We have developed an improved method for solid-phase coupling of ligands to oligonucleotides via disulfides directly after solid-phase synthesis. A 2'-thiol introduced using a modified nucleotide building block was orthogonally deprotected on the controlled pore glass solid support with N-butylphosphine. Oligolysine peptides and a short monodisperse ethylene glycol chain were successfully coupled to the deprotected thiol. Cleavage from the resin and full removal of oligonucleotide protection groups were achieved using methanolic ammonia. After standard desalting, and without further purification, homogenous conjugates were obtained as demonstrated by HPLC, gel electrophoresis, and mass spectrometry. The attachment of both amphiphilic and cationic ligands proves the versatility of the conjugation procedure. An antisense oligonucleotide conjugate with hexalysine showed pronounced gene silencing in a cell culture tumor model in the absence of a transfection reagent and the corresponding ethylene glycol conjugate resulted in down regulation of the target gene to nearly 50% after naked application.
Amide bond-containing monodisperse polyethylene glycols beyond 10 000 Da.
Wan Zihong,Li Yu,Bo Shaowei,Gao Ming,Wang Xuemeng,Zeng Kai,Tao Xin,Li Xuefei,Yang Zhigang,Jiang Zhong-Xing
Organic & biomolecular chemistry
Although monodisperse polyethylene glycols (M-PEGs) above 4000 Da are especially valuable in biomedical applications, their synthesis remains a long-standing challenge. To this end, a peptide-based strategy for such M-PEGs was developed. With macrocyclic sulfates as the key intermediates, a panel of oligoethylene glycol (OEG) containing ω-amino acids were prepared with high efficiency. Through solid phase peptide synthesis (SPPS), these amino acids were conveniently assembled into a series of amide bond-containing M-PEGs with high flexibility in molecular weight and amide density selection. With this strategy, an M-PEG of 10 262 Da was prepared on a gram scale and its biocompatibility was assessed in a mice model.