Morphologic Design of Silver-Bearing Sugar-Based Polymer Nanoparticles for Uroepithelial Cell Binding and Antimicrobial Delivery.

Platelet-like and cylindrical nanostructures from sugar-based polymers are designed to mimic the aspect ratio of bacteria and achieve uroepithelial cell binding and internalization, thereby improving their potential for local treatment of recurrent urinary tract infections. Polymer nanostructures, derived from amphiphilic block polymers composed of zwitterionic poly(d-glucose carbonate) and semicrystalline poly(l-lactide) segments, were constructed with morphologies that could be tuned to enhance uroepithelial cell binding. These nanoparticles exhibited negligible cytotoxicity, immunotoxicity, and cytokine adsorption, while also offering substantial silver cation loading capacity, extended release, and in vitro antimicrobial activity (as effective as free silver cations) against uropathogenic Escherichia coli. In comparison to spherical analogues, cylindrical and platelet-like nanostructures engaged in significantly higher association with uroepithelial cells, as measured by flow cytometry; despite their larger size, platelet-like nanostructures maintained the capacity for cell internalization. This work establishes initial evidence of degradable platelet-shaped nanostructures as versatile therapeutic carriers for treatment of epithelial infections.

In Situ Production of Ag/Polymer Asymmetric Nanoparticles via a Powerful Light-Driven Technique.

As a rapid, controllable, and easily transferrable approach to the preparation of antimicrobial nanoparticle systems, a one-step, light-driven procedure was developed to produce asymmetric hybrid inorganic-organic nanoparticles (NPs) directly from a homogeneous Ag/polymer mixture. An amphiphilic triblock polymer was designed and synthesized to build biocompatible NPs, consisting of poly(ethylene oxide) (PEO), carboxylic acid-functionalized polyphosphoester (PPE), and poly(l-lactide) (PLLA). Unexpectedly, snowman-like asymmetric nanostructures were subsequently obtained by simply loading silver cations into the polymeric micelles together with purification via centrifugation. With an understanding of the chemistry of the asymmetric NP formation, a controllable preparation strategy was developed by applying UV irradiation. A morphology transition was observed by transmission electron microscopy over the UV irradiation time, from small silver NPs distributed inside the micelles into snowman-like asymmetric NPs, which hold promise for potential antimicrobial applications with their unique two-stage silver release profiles.

Minocycline and Silver Dual-Loaded Polyphosphoester-Based Nanoparticles for Treatment of Resistant Pseudomonas aeruginosa.

Pseudomonas aeruginosa has been detected in the lungs of ∼50% of patients with cystic fibrosis (CF), including 20% of adult CF patients. The majority of these adult patients harbor multi-drug resistant (MDR) strains, limiting the available treatment options. Silver has long been used as a broad-spectrum antimicrobial agent with a low incidence of resistance. Despite low toxicity, poor availability of silver cations mandates a high dosage to effectively eradicate infections. To address this shortcoming of silver, nanoparticles have been used as delivery devices to improve treatment outcomes. Furthermore, studies have demonstrated that synergistic combinations with careful dose calibrations and efficient delivery systems result in superior antimicrobial activity while avoiding potential side effects of both therapeutics. Here 4-epi-minocycline, a metabolite of minocycline, was identified as an active antimicrobial against P. aeruginosa using a high-throughput screen. The antimicrobial activities of 4-epi-minocycline, minocycline, and silver acetate against clinical isolates of P. aeruginosa obtained from CF patients were evaluated in vitro. Next, the synergistic activity of the silver/minocycline combination against P. aeruginosa isolates was investigated using checkerboard assays and identified with end-point colony forming unit determination assays. Finally, nanoparticles coloaded with minocycline and silver were evaluated in vitro for antimicrobial activity. The results demonstrated that both silver and minocycline are potent antimicrobials alone and that the combination allows a reduced dosage of both therapeutics to achieve the same antimicrobial effect. Furthermore, the proposed synergistic silver/minocycline combination can be coloaded into nanoparticles as a next-generation antibiotic to combat the threats presented by MDR pathogens.

Functional, Degradable Zwitterionic Polyphosphoesters as Biocompatible Coating Materials for Metal Nanostructures.

A zwitterionic polyphosphoester (zPPE), specifically l-cysteine-functionalized poly(but-3-yn-1-yloxy)-2-oxo-1,3,2-dioxaphospholane (zPBYP), has been developed as a poly(ethylene glycol) (PEG) alternative coating material for gold nanoparticles (AuNPs), the most extensively investigated metal nanoparticulate platform toward molecular imaging, photothermal therapy, and drug delivery applications. Thiol-yne conjugation of cysteine transformed an initial azido-terminated and alkynyl-functionalized PBYP homopolymer into zPBYP, offering hydrolytic degradability, biocompatibility, and versatile reactive moieties for installation of a range of functional groups. Despite minor degradation during purification, zPPEs were able to stabilize AuNPs presumably through multivalent interactions between combinations of the side chain zwitterions (thioether and phosphoester groups of the zPPEs with the AuNPs). 31P NMR studies in D2O revealed ca. 20% hydrolysis of the phosphoester moieties of the repeat units had occurred during the workup and purification by aqueous dialysis at pH 3 over ca. 1 d, as observed by the 31P signal of the phosphotriesters resonating at ca. -0.5 to -1.7 shifting downfield to ca. 1.1 to -0.4 ppm, attributed to transformation to phosphates. Further hydrolysis of side chain and backbone units proceeded to an extent of ca. 75% over the next 2 d in nanopure water (pH 5-6). The NMR degradation results were consistent with the broadening and red-shift of the surface plasmon resonance (SPR) observed by UV-vis spectroscopy of the zPPE-coated AuNPs in water over time. All AuNP formulations in this study, including those with citrate, PEG, and zPPE coatings, exhibited negligible immunotoxicity, as determined by cytokine overexpression in the presence of the nanostructures relative to those in cell culture medium. Notably, the zPPE-coated AuNPs displayed superior antifouling properties, as assessed by the extent of cytokine adsorption relative to both the PEGylated and citrate-coated AuNPs. Taken together, the physicochemical and biological evaluations of zPPE-coated AuNPs in conjunction with PEGylated and citrate-coated analogues indicate the promise of zPPEs as favorable alternatives to PEG coatings, with negligible immunotoxicity, good antifouling performance, and versatile reactive groups that enable the preparation of highly tailored nanomaterials for diverse applications.

Advancing the Development of Highly-Functionalizable Glucose-Based Polycarbonates by Tuning of the Glass Transition Temperature.

Fundamental studies that gain an understanding of the tunability of physical properties of natural product-based polymers are vital for optimizing their performance in extensive applications. Variation of glass transition temperature ( Tg) was studied as a function of the side chain structure and molar mass for linear poly(glucose carbonate)s. A remarkable range of Tg values, from 38 to 125 °C, was accomplished with six different alkyloxycarbonyl side chains. The impact of molar mass on Tg was investigated for two series of polymers and discrete oligomers synthesized and fractionated with precise control over the degrees of polymerization. The Tg was found to be greatly influenced by a synergistic effect of the flexibility and bulkiness of the repeating unit side chain, as well as the chain end relative free volume. This work represents an important advance in the development of glucose-based polycarbonates, as materials that possess high degrees of functionalizability to be capable of exhibiting diversified physicochemical and thermal properties by simple side chain modification.

Chemical Design of Both a Glutathione-Sensitive Dimeric Drug Guest and a Glucose-Derived Nanocarrier Host to Achieve Enhanced Osteosarcoma Lung Metastatic Anticancer Selectivity.

Although nanomedicines have been pursued for nearly 20 years, fundamental chemical strategies that seek to optimize both the drug and drug carrier together in a concerted effort remain uncommon yet may be powerful. In this work, two block polymers and one dimeric prodrug molecule were designed to be coassembled into degradable, functional nanocarriers, where the chemistry of each component was defined to accomplish important tasks. The result is a poly(ethylene glycol) (PEG)-protected redox-responsive dimeric paclitaxel (diPTX)-loaded cationic poly(d-glucose carbonate) micelle (diPTX@CPGC). These nanostructures showed tunable sizes and surface charges and displayed controlled PTX drug release profiles in the presence of reducing agents, such as glutathione (GSH) and dithiothreitol (DTT), thereby resulting in significant selectivity for killing cancer cells over healthy cells. Compared to free PTX and diPTX, diPTX@CPGC exhibited improved tumor penetration and significant inhibition of tumor cell growth toward osteosarcoma (OS) lung metastases with minimal side effects both in vitro and in vivo, indicating the promise of diPTX@CPGC as optimized anticancer therapeutic agents for treatment of OS lung metastases.

Multi-responsive polypeptide hydrogels derived from N-carboxyanhydride terpolymerizations for delivery of nonsteroidal anti-inflammatory drugs.

A polypeptide-based hydrogel system, when prepared from a diblock polymer with a ternary copolypeptide as one block, exhibited thermo-, mechano- and enzyme-responsive properties, which enabled the encapsulation of naproxen (Npx) during the sol-gel transition and its release in the gel state. Statistical terpolymerizations of l-alanine (Ala), glycine (Gly) and l-isoleucine (Ile) NCAs at a 1 : 1 : 1 feed ratio initiated by monomethoxy monoamino-terminated poly(ethylene glycol) afforded a series of methoxy poly(ethylene glycol)-block-poly(l-alanine-co-glycine-co-l-isoleucine) (mPEG-b-P(A-G-I)) block polymers. ß-Sheets were the dominant secondary structures within the polypeptide segments, which facilitated a heat-induced sol-to-gel transition, resulting from the supramolecular assembly of ß-sheets into nanofibrils. Deconstruction of the three-dimensional networks by mechanical force (sonication) triggered the reverse gel-to-sol transition. Certain enzymes could accelerate the breakdown of the hydrogel, as determined by in vitro gel weight loss profiles. The hydrogels were able to encapsulate and release Npx over 6 days, demonstrating the potential application of these polypeptide hydrogels as an injectable local delivery system for small molecule drugs.

Improving paclitaxel delivery: in vitro and in vivo characterization of PEGylated polyphosphoester-based nanocarriers.

Nanomaterials have great potential to offer effective treatment against devastating diseases by providing sustained release of high concentrations of therapeutic agents locally, especially when the route of administration allows for direct access to the diseased tissues. Biodegradable polyphosphoester-based polymeric micelles and shell cross-linked knedel-like nanoparticles (SCKs) have been designed from amphiphilic block-graft terpolymers, PEBP-b-PBYP-g-PEG, which effectively incorporate high concentrations of paclitaxel (PTX). Well-dispersed nanoparticles physically loaded with PTX were prepared, exhibiting desirable physiochemical characteristics. Encapsulation of 10 wt% PTX, into either micelles or SCKs, allowed for aqueous suspension of PTX at concentrations up to 4.8 mg/mL, as compared to <2.0 µg/mL for the aqueous solubility of the drug alone. Drug release studies indicated that PTX released from these nanostructures was defined through a structure-function relationship, whereby the half-life of sustained PTX release was doubled through cross-linking of the micellar structure to form SCKs. In vitro, physically loaded micellar and SCK nanotherapeutics demonstrated IC50 values against osteosarcoma cell lines, known to metastasize to the lungs (CCH-OS-O and SJSA), similar to the pharmaceutical Taxol formulation. Evaluation of these materials in vivo has provided an understanding of the effects of nanoparticle structure-function relationships on intratracheal delivery and related biodistribution and pharmacokinetics. Overall, we have demonstrated the potential of these novel nanotherapeutics toward future sustained release treatments via administration directly to the sites of lung metastases of osteosarcoma.

Tetrahydroindazolone substituted 2-aminobenzamides as fluorescent probes: switching metal ion selectivity from zinc to cadmium by interchanging the amino and carbamoyl groups on the fluorophore.

Three fluorescent probes CdABA', CdABA and ZnABA', which are structural isomers of ZnABA, have been designed with N,N-bis(2-pyridylmethyl) ethylenediamine (BPEA) as chelator and 2-aminobenzamide as fluorophore. These probes can be divided into two groups: CdABA, CdABA' for Cd(2+) and ZnABA, ZnABA' for Zn(2+). Although there is little difference in their chemical structures, the two groups of probes exhibit totally different fluorescence properties for preference of Zn(2+) or Cd(2+). In the group of Zn(2+) probes, ZnABA/ZnABA' distinguish Zn(2+) from Cd(2+) with F(Zn)(2+)-F(Cd)(2+) = 1.87-2.00. Upon interchanging the BPEA and carbamoyl groups on the aromatic ring of the fluorophore, the structures of ZnABA/ZnABA' are converted into CdABA/CdABA'. Interestingly, the metal ions selectivity of CdABA/CdABA' was switched to discriminate Cd(2+) from Zn(2+) with F(Cd)(2+)-F(Zn)(2+) = 2.27-2.36, indicating that a small structural modification could lead to a remarkable change of the metal ion selectivity. (1)H NMR titration and ESI mass experiments demonstrated that these fluorescent probers exhibited different coordination modes for Zn(2+) and Cd(2+). With CdABA' as an example, generally, upon addition of Cd(2+), the fluorescence response possesses PET pathway to display no obvious shift of maximum λ(em) in the absence or presence of Cd(2+). However, an ICT pathway could be employed after adding Zn(2+) into the CdABA' solution, resulting in a distinct red-shift of maximal λ(em).

Experiments and Simulations of Complex Sugar-Based Coil-Brush Block Polymer Nanoassemblies in Aqueous Solution.

In this work, we investigated the fundamental molecular parameters that guide the supramolecular assembly of glucose-based amphiphilic coil-brush block polymers in aqueous solution and elucidated architecture-morphology relationships through experimental and simulation tools. Well-defined coil-brush polymers were synthesized through ring-opening polymerizations (ROP) of glucose carbonates to afford norbornenyl-functionalized poly(glucose carbonate) (NB-PGC) macromonomers, followed by sequential ring-opening metathesis polymerizations (ROMP) of norbornene N-hydroxysuccinimidyl (NHS) esters and the NB-PGC macromonomers. Variation of the macromonomer length and grafting through ROMP conditions allowed for a series of coil-brush polymers to be synthesized with differences in the brush and coil dimensions, independently, where the side chain graft length and brush backbone were used to tune the brush, and the coil block length was used to vary the coil. Hydrolysis of the NHS moieties gave the amphiphilic coil-brush polymers, where the hydrophilic-hydrophobic ratios were dependent on the brush and coil relative dimensions. Experimental assembly in solution was studied and found to yield a variety of structurally dependent nanostructures. Simulations were conducted on the solution assembly of coil-brush polymers, where the polymers were represented by a coarse-grained model and the solvent was represented implicitly. There is qualitative agreement in the phase diagrams obtained from simulations and experiments, in terms of the morphologies of the assembled nanoscopic structures achieved as a function of coil-brush design parameters ( e.g., brush and coil lengths, composition). The simulations further showed the chain conformations adopted by the coil-brush polymers and the packing within these assembled nanoscopic structures. This work enables the predictive design of nanostructures from this glucose-based coil-brush polymer platform while providing a fundamental understanding of interactions within solution assembly of complex polymer building blocks.

Assessment of Copper Nanoclusters for Accurate in Vivo Tumor Imaging and Potential for Translation.

Nanoparticles have been widely used for preclinical cancer imaging. However, their successful clinical translation is largely hampered by potential toxicity, unsatisfactory detection of malignancy at early stages, inaccurate diagnosis of tumor biomarkers, and histology for imaging-guided treatment. Herein, a targeted copper nanocluster (CuNC) is reported with high potential to address these challenges for future translation. Its ultrasmall structure enables efficient renal/bowel clearance, minimized off-target effects in nontargeted organs, and low nonspecific tumor retention. The pH-dependent in vivo dissolution of CuNCs affords minimal toxicity and potentially selective drug delivery to tumors. The intrinsic radiolabeling through the direct addition of 64Cu to CuNC (64Cu-CuNCs-FC131) synthesis offers high specific activity for sensitive and accurate detection of CXCR4 via FC131-directed targeting in novel triple negative breast cancer (TNBC) patient-derived xenograft mouse models and human TNBC tissues. In summary, this study not only reveals the potential of CXCR4-targeted 64Cu-CuNCs for TNBC imaging in clinical settings, but also provides a useful strategy to design and assess the translational potential of nanoparticles for cancer theranostics.

Acid-Triggered Polymer Backbone Degradation and Disassembly to Achieve Release of Camptothecin from Functional Polyphosphoramidate Nanoparticles.

Camptothecin (CPT) is a promising anticancer drug, yet its therapeutic potential has been limited by poor water solubility and facile hydrolysis of the lactone form into an inactive carboxylate form at neutral pH. In this work, a fundamental synthetic methodology was advanced to allow for the preparation of well-defined functional polyphosphoramidate (PPA)-based block copolymers that coassembled with CPT into nanoparticles, which underwent coincident acid-triggered polymer backbone degradation, nanoparticle disassembly, and CPT release. Encapsulation of CPT by the PPA polymer inhibited premature hydrolysis of CPT at pH 7.4 and enabled accelerated CPT release at pH 5.0 (ca. 4× faster than at pH 7.4). Two degradable oxazaphospholidine monomers, with one carrying an alkyne group, were synthesized to access well-defined block PPAs (dispersity, D<1.2) via sequential organobase-catalyzed ring-opening polymerizations (ROP). The resulting amphiphilic block copolymers (PEOMP-b-PBYOMP) were physically loaded with CPT to achieve well-dispersed nanotherapeutics, which allowed the aqueous suspension of CPT at concentrations up to 3.2 mg/mL, significantly exceeding the aqueous solubility of the drug (<2.0 µg/mL at 37 °C). Cytotoxicity studies revealed enhanced efficacy of the CPT-loaded nanoparticles over free CPT in cancer cells and similar toxicity in normal cells.

Polyphosphoramidates That Undergo Acid-Triggered Backbone Degradation.

The direct and facile synthesis of polyphosphoramidates (PPAs) with acid-labile phosphoramidate backbone linkages are reported, together with demonstration of their hydrolytic degradability, evaluated under acidic conditions. The introduction of acid-labile linkages along the polymer backbone led to rapid degradation of the polymer backbone dependent upon the environmental stimuli. An oxazaphospholidine monomer bearing a phosphoramidate linkage was designed and synthesized to afford the PPAs via organobase-catalyzed ring-opening polymerization in a controlled manner. The hydrolytic degradation of the PPAs was studied, revealing breakdown of the polymer backbone through cleavage of the phosphoramidate linkages under acidic conditions.

Polyphosphoester nanoparticles as biodegradable platform for delivery of multiple drugs and siRNA.

Delivery of multiple therapeutics and/or diagnostic agents to diseased tissues is challenging and necessitates the development of multifunctional platforms. Among the various strategies for design of multifunctional nanocarriers, biodegradable polyphosphoester (PPE) polymers have been recently synthesized via a rapid and simple synthetic strategy. In addition, the chemical structure of the polymer could be tuned to form nanoparticles with varying surface chemistries and charges, which have shown exceptional safety and biocompatibility as compared to several commercial agents. The purpose of this study was to exploit a mixture of PPE nanoparticles of cationic and neutral surface charges for multiple delivery of anticancer drugs (ie, sorafenib and paclitaxel) and nucleic acids (ie, siRNA). Cationic PPE polymers could efficiently complex siRNA, and the stability of the nanoparticles could be maintained in physiological solutions and upon freeze-drying and were able to deliver siRNA in vivo when injected intravenously in mice. Commercially available cationic polyethylenimine polymer had LD50 of ca. 61.7 mg/kg in mice, whereas no animal died after injection of the cationic PPE polymer at a dose of >130 mg/kg. Neutral PPE nanoparticles were able to encapsulate two hydrophobic drugs, namely, sorafenib and paclitaxel, which are commonly used for the treatment of hepatocellular carcinoma. Mixing the neutral and cationic PPE nanoparticles did not result in any precipitation, and the size characteristics of both types of nanoparticles were maintained. Hence, PPE polymers might have potential for the delivery of multiple drugs and diagnostic agents to diseased tissues via simple synthesis of the individual polymers and assembly into nanoparticles that can host several drugs while being mixed in the same administration set, which is of importance for industrial and clinical development.

Design and development of multifunctional polyphosphoester-based nanoparticles for ultrahigh paclitaxel dual loading.

Multifunctional polyphosphoester-based nanoparticles capable of loading paclitaxel (PTX) both chemically and physically were prepared, achieving an ultrahigh equivalent PTX aqueous concentration of 25.30 mg mL-1. The dual-loaded nanoparticles were effective in killing cancer cells, which has the potential to minimize the amount of nanocarriers needed for clinical applications, due to their ultrahigh loading capacity.

Degradable polyphosphoester-based silver-loaded nanoparticles as therapeutics for bacterial lung infections.

In this study, a new type of degradable polyphosphoester-based polymeric nanoparticle, capable of carrying silver cations via interactions with alkyne groups, has been developed as a potentially effective and safe treatment for lung infections. It was found that up to 15% (w/w) silver loading into the nanoparticles could be achieved, consuming most of the pendant alkyne groups along the backbone, as revealed by Raman spectroscopy. The well-defined Ag-loaded nanoparticles released silver in a controlled and sustained manner over 5 days, and displayed enhanced in vitro antibacterial activities against cystic fibrosis-associated pathogens and decreased cytotoxicity to human bronchial epithelial cells, in comparison to silver acetate.

Construction of a versatile and functional nanoparticle platform derived from a helical diblock copolypeptide-based biomimetic polymer.

Sequential polymerization of N-carboxyanhydrides accelerated by nitrogen flow is utilized to generate a novel well-defined diblock copolypeptide (PDI = 1.08), with incorporation of alkyne-functionalized side-chain groups allowing for rapid and efficient thiol-yne click-type modifications, followed by self-assembly into nanopure water to construct a helical polypeptide-based versatile and functional nanoparticle platform.

Multi-responsive Hydrogels Derived from the Self-assembly of Tethered Allyl-functionalized Racemic Oligopeptides.

A multi-responsive triblock hydrogelator oligo(dl-allylglycine)-block-poly(ethylene glycol)-block-oligo(dl-allylglycine) (ODLAG-b-PEG-b-ODLAG) was synthesized facilely by ring-opening polymerization (ROP) of DLAG N-carboxyanhydride (NCA) with a diamino-terminated PEG as the macroinitiator. This system exhibited heat-induced sol-to-gel transitions and either sonication- or enzyme-induced gel-to-sol transitions. The ß-sheeting of the oligopeptide segments was confirmed by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and wide-angle X-ray scattering (WAXS). The ß-sheets further displayed tertiary ordering into fibrillar structures that, in turn generated a porous and interconnected hydrogel matrix, as observed via transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The reversible macroscopic sol-to-gel transitions triggered by heat and gel-to-sol transitions triggered by sonication were correlated with the transformation of nanostructural morphologies, with fibrillar structures observed in gel and spherical aggregates in sol, respectively. The enzymatic breakdown of the hydrogels was also investigated. This allyl-functionalized hydrogelator can serve as a platform for the design of smart hydrogels, appropriate for expansion into biological systems as bio-functional and bio-responsive materials.

Tunable stereoselectivity during sialylation using an N-acetyl-5-N,4-O-oxazolidinone-protected p-toluene 2-thio-sialoside donor with Tf(2)O/Ph(2)SO/TTBPy.

An N-acetyl-5-N,4-O-oxazolidinone-protected p-toluene 2-thio-sialoside donor, promoted by Ph(2)SO/Tf(2)O/TTBPy, was thoroughly investigated in the coupling to various acceptors. The stereoselectivity of the sialylation was found to be dependent on the various reaction conditions, such as pre-activation time, reaction time, the amount of Ph(2)SO, and TTBPy. A detailed Ph(2)SO/Tf(2)O-promoted glycosylation mechanism was proposed, which contained three crucial reactive species: an oxacarbenium ion, C2-sialyloxosulfonium salts, and oxosulfonium supramers. Our research results indicate that it is possible to tune the stereoselectivity of the sialylation by carefully changing the reaction conditions. For instance, Ph(2)SO (2.0-3.0 equiv)/TTBPy (0-1.0 equiv) promotion gives higher α-selective sialylation in dichloromethane, while Ph(2)SO (4-5 equiv)/TTBPy (0 equiv) or Ph(2)SO (2.0 equiv)/TTBPy (2.0 equiv) affords lower stereoselectivity.