Fabrication of Dual pH/Reduction-Responsive P(CL-co-ACL)-Based Cross-Linked Polymeric Micelles for PTX Delivery.

To enhance the stability of the polymeric micelles and optimize their drug-controlled release ability, three disulfide-linked polyethylene glycol methyl ether methacrylate-disulfide-poly(ε-caprolactone-co-γ-amine-ε-caprolactone) (PPEGMA-SS-P(CL-co-ACL)) polymers were synthesized and characterized by 1H NMR, GPC, and FT-IR successfully, and their dual pH/reduction-responsive cross-linked polymeric micelles were prepared for paclitaxel (PTX) delivery by using 2,3-dimethylmaleic anhydride (DMMA) as the cross-linking agent. The PTX loading capacity (LC) and encapsulation efficiency (EE) values of the cross-linked micelles formed by PPEGMA8-SS-P(CL47-co-ACL15) achieved were 23.96% and 71.58%, slightly higher than those of un-cross-linked micelles. Both particle sizes of blank micelles and in vitro drug release of PTX-loaded micelles confirmed that compared with those un-cross-linked micelles, the cross-linked micelles were more stable at pH 7.4 + 0 mM DTT, with a PTX cumulative release of 13% at 120 h, while the PTX cumulative release of the cross-linked micelles at pH 5.0 + 10 mM DTT were close to that of un-cross-linked micelles after 60 h, indicating the successful reversible cross-linking and smooth drug release of the cross-linked micelles. The cytotoxicity assay showed that PPEGMA8-SS-P(CL47-co-ACL15) and its cross-linked micelles had low cell cytotoxicity, and both PTX-loaded micelles revealed a certain inhibitory effect on HepG2 cells. These results revealed that the dual pH/reduction-responsive cross-linked polymeric micelles prepared from PPEGMA8-SS-P(CL47-co-DCL15) were a promising candidate for PTX delivery.

Folate modified dual pH/reduction-responsive mixed micelles assembled using FA-PEG-PDEAEMA and PEG-SS-PCL for doxorubicin delivery.

Aiming at achieving the concurrent performances of high loading, well controlled release and active targeted delivery, folate (FA) modified dual pH/reduction-responsive mixed polymeric micelles were rationally assembled using FA-PEG-PDEAEMA and PEG-SS-PCL by dissipative particle dynamics (DPD) simulations. The optimized polymers PEG112-PDEAEMA40, FA-PEG112-PDEAEMA40, and PEG112-SS-PCL70 were synthesized and characterized using 1H NMR, FT-IR and GPC, and their mixed micelles were applied for doxorubicin (DOX) delivery. The drug loading capacity (LC) and encapsulation efficiency (EE) values of the MIX1 (FA-PEG112-PDEAEMA40/PEG112-SS-PCL70) at a DOX/polymer feeding ratio of 15 mg/30 mg were 20.22% and 50.69%, which were higher than those of single polymer micelles and MIX2 (PEG112-PDEAEMA40/PEG112-SS-PCL70). Particle size distributions, mesoscopic morphologies, DPD simulations and in vitro drug release profiles all confirmed the well-controlled release performance of the DOX-loaded micelles formed by MIX1: slow DOX release with a cumulative release of 20.46% in the neutral environment and accelerated release with a cumulative release of 74.20% at pH 5.0 + 10 mM DTT within 120 h, which were similar to those of MIX2. Cytotoxicity assay found that both MIX1 and MIX2 blank micelles were biocompatible, and a superior inhibitory effect of the FA-modified DOX-loaded micelles MIX1 on HepG2 cells was found compared to that of free DOX and non-FA-modified DOX-loaded micelles MIX2. All of these confirmed the superiority of MIX1 micelles with high loading capacity, well controlled release, and enhanced inhibitory effects on HepG2 cells, which might be a prospective candidate for anticancer drug delivery.

Systematic Design and Study of Star-like Polymeric Prodrug Unimolecular Micelles ß-CD-P[CL-co-(ACL-g-DOX)-SS-MPEG]21 by DPD Simulations.

Unimolecular micelles composed of a single polymeric molecule have recently attracted significant attention in anti-cancer drug delivery due to their high thermodynamic stability and small particle sizes. Applying the prodrug strategy to unimolecular micelles may provide superior nano-drug carriers with simultaneous high stability, low drug leakage, and well-drug loading capacity. However, the formation mechanism of the unimolecular prodrug micelles, the superiority of the prodrug strategy, as well as the prodrug controlled release mechanism were scantily understood at the mesoscopic scale. In this work, dissipative particle dynamics mesoscopic simulations were employed to investigate the self-assembly behavior, formation conditions, drug distribution regularities, and the prodrug release process of the star-like polymeric prodrug unimolecular micelles formed by ß-CD-P[CL-co-(ACL-g-DOX)-SS-MPEG]21. A special bond-breaking script was used to accomplish the bond-breaking simulation of the grafted DOX bonds and the disulfide bonds. Results showed that to form well monodispersed and superior DOX-loaded unimolecular micelles, the polymer concentration should be well controlled at low volume fractions (≤10.59%), and the detailed molecular structure of the polymer was suggested as ß-cyclodextrin-P[caprolactone-co-(amino caprolactone-g-doxorubicin)-disulfide-methyl polyethylene glycol]21) (ß-CD-P[CL30-co-(ACL-g-DOX)8-SS-MPEG49]21). By comparison with the DOX physically loaded micelles, it was found that the prodrug unimolecular micelles with DOX grafted on the polymer displayed no drug leakage and superior drug loading capacity. Simulations on the prodrug release process showed that the prodrug unimolecular micelles assembled by ß-CD-P[CL30-co-(ACL-g-DOX)8-SS-MPEG49]21 would provide good dual pH/reduction-responsive DOX release performance.

Mesoscopic Simulations of Diselenide-Containing Crosslinked Doxorubicin-Loaded Micelles and Their Tumor Microenvironment Responsive Release Behaviors.

There is currently limited research on the structure-property relationship of reduction stimuli-responsive polymeric crosslinked micelles using mesoscopic simulations. Herein, dissipative particle dynamics (DPD) simulations were used to simulate the self-assembly process of the blank non-crosslinked micelle, the structure and doxorubicin (DOX) distribution of diselenide crosslinked micelle with different crosslinker contents (CCs) based on the nearest-neighbor bonding principle. The results revealed that the formation of a three-layer spherical micelle and the loaded DOX mainly distributed in the polycaprolactone (PCL) core and hydroxyethyl methacrylate (HEMA) mesosphere. The larger the dosage of DOX, the more DOX encapsulated, but the encapsulation of DOX in the hydrophobic domain would reach saturation when the dosage increased to 6.0 %. In micelles with lower CCs or crosslinking levels (CLs), DOX entered the middle layer and the inner core faster. Then, based on the nearest media-bead bond breaking principle and subsequently DPD simulation, the effects of different CCs on the micelle structure and DOX release properties were investigated. Low CC could cause fast drug release. With the increase of CCs, the micelle showed a slower DOX release trend. The multilayer crosslinked network system also affected the DOX release rate. Hence, this work can provide some mesoscale guidance for the structural design and structure-property relationship of stimuli-responsive reversible crosslinked micelles for drug delivery.

DPD simulations on mixed polymeric DOX-loaded micelles assembled from PCL-SS-PPEGMA/PDEA-PPEGMA and their dual pH/reduction-responsive release.

The design of mixed polymeric micelles by a combination of two or more dissimilar polymers is a potential strategy to achieve multiple stimuli-response for anti-cancer drug delivery. However, their drug loading co-micellization behavior and multiple stimuli-responsive drug release mechanism have been poorly understood at the mesoscopic level, especially in the system that involves reduction-response due to the difficulty of simulation on the cleavage of chemical bonds. In this work, the co-micellization behavior, drug distribution regularities and dual pH/reduction-responsive drug release process of mixed micelles formed by disulfide-linked polycaprolactone-b-polyethylene glycol methyl ether methacrylate (PCL-SS-PPEGMA) and poly(ethylene glycol) methyl ether-b-poly(N,N-diethylamino ethyl methacrylate) (PDEA-PPEGMA) were studied by dissipative particle dynamics (DPD) mesoscopic simulations. A dedicated bond-breaking script was employed to accomplish the disulfide bond-breaking simulations. The results showed that PCL55-SS-PPEGMA10 and PDEA34-PPEGMA11 could be well mixed to form superior DOX-loaded micelles with good drug-loading capacity and drug-controlled release performance. To prepare the DOX-loaded micelles with optimized properties, the simulation results suggested the feed ratio of DOX:PCL55-SS-PPEGMA10:PDEA34-PPEGMA11 set to 3:4:4. Compared with the two single stimuli-response, the dual pH/reduction-response process perfectly combined both pH-response and reduction-response together, providing a higher release rate of DOX. Therefore, this study provides theoretical guidance aimed at the property optimization and micellar structure design of the dual pH/reduction-responsive mixed micelles.

DPD simulations and experimental study on reduction-sensitive polymeric micelles self-assembled from PCL-SS-PPEGMA for doxorubicin controlled release.

Delivery of anticancer drugs by amphiphilic polymeric micelles with disulfide bonds as the reduction-responsive groups has potential application in the field of drug-controlled release. In this study, three disulfide-linked polycaprolactone-b-polyethylene glycol methyl ether methacrylate (PCL-SS-PPEGMA) were synthesized and confirmed by 1H NMR and GPC, and then used for doxorubicin (DOX) delivery. The CMC values of the three PCL-SS-PPEGMA micelles were low (0.71-4.56 mg/L), indicative of the good stability of micelles in aqueous solution. The drug loading content (LC) and encapsulation efficiency (EE), together with the DOX accelerated release profiles were determined, with good drug loading capacity and well drug-controlled release performance. And to explore the mesoscopic behavior of reduction-responsive drug-loaded polymeric micelles, by using a dedicated disulfide bond-breaking model and script, dissipative particle dynamics (DPD) simulations were carried out on the three PCL-SS-PPEGMA polymers. Their self-assembled behavior, formation of DOX-loaded micelles, the disulfide bond-breaking process, as well as the DOX reduction-responsive release process were simulated and assessed. Comparing the DPD simulation results with the experimental data, we found that they were in good agreement, effectively demonstrating that the DPD simulation method developed can provide a practical mesoscopic approach for the reduction-responsive drug-loaded polymeric micelles that involved the cleavage of dynamic covalent bonds.

pH-Sensitive Mixed Micelles Assembled from PDEAEMA-PPEGMA and PCL-PPEGMA for Doxorubicin Delivery: Experimental and DPD Simulations Study.

To decrease critical micelle concentration (CMC), improve stability, and keep high drug-loading capacity, three pH-sensitive mixed micelles applied for anticancer drug controlled delivery were prepared by the mixture of polymers poly (N,N-diethylaminoethyl methacrylate)-b-poly(poly(ethylene glycol) methyl ether methacrylate) (PDEAEMA-PPEGMA) and polycaprolactone-b-poly (poly(ethylene glycol) methyl ether methacrylate) (PCL-PPEGMA), which were synthesized and confirmed by 1H NMR and gel permeation chromatographic (GPC). The critical micelle concentration (CMC) values of the prepared mixed micelles were low, and the micellar sizes and zeta potentials of the blank mixed micelles demonstrated good pH-responsive behavior. Combined experimental techniques with dissipative particle dynamics (DPD) simulation, the particle sizes, zeta potentials, drug loading content (LC), encapsulation efficiency (EE), aggregation morphologies, and doxorubicin (DOX) distribution of the mixed micelles were investigated, and the high DOX-loading capacity of the mixed micelles was found. Both in vitro DOX release profiles and DPD simulations of the DOX dynamics release process exhibited less leakage and good stability in neutral conditions and accelerated drug release behavior with a little initial burst in slightly acidic conditions. Cytotoxicity tests showed that the polymer PDEAEMA-PPEGMA and the blank mixed micelles had good biocompatibility, and DOX-loaded mixed micelles revealed certain cytotoxicity. These results suggest that the drug-loaded mixed micelles that consisted of the two polymers PDEAEMA-PPEGMA and PCL-PPEGMA can be new types of pH-responsive well-controlled release anticancer drug delivery mixed micelles.

Mesoscopic simulations of drug-loaded diselenide crosslinked micelles: Stability, drug loading and release properties.

Intelligent reversible crosslinked micelles that have a good balance of structure stability in normal tissue and controlled drug release responded to the tumor microenvironment are highly promising novel drug delivery systems. However, to date, there have been very few reports about mesoscale simulations of drug-loaded polymeric reversible crosslinked micelles. Here, dissipative particle dynamics (DPD) simulation, the nearest-neighbor bonding principle, and the nearest media-bead bond breaking principle were used to investigate the influence of physiological environment along with low tumor pH and reduction microenvironment on the stability and doxorubicin (DOX) distribution of the star polymer [PCL-b-P(HEMA-Se-Se˜)-b-PPEGMA]6 diselenide crosslinked micelles with different diselenide crosslinking levels (CLs). The self-assembly process results obtained by DPD simulations reveal the formation of three-layer spherical micelles with the loaded DOX mainly distributed at the interfacial regions of the inner PCL core and middle HEMA layer. The structural stability and DOX loading capacity of the micelles can be improved by appropriately increasing the CL based on the nearest-neighbor bonding principle due to the effect of the pressure exerted by the crosslink that squeezes the loaded drugs from the intermediate and interfacial layers into the micelle core. Furthermore, the effect of breaking of the diselenide bond on the drug release properties was investigated through the use of the nearest media-bead bond breaking principle. A low CL gives rise to intense drug release, increasing the toxic side effects on the system. With the increase in the CL, the micelles show the transformation from local crosslinking to compact crosslinking, leading to slower drug release. Therefore, this work can provide some guidance on the mesoscale for the structural design and controlled construction of reversible crosslinked micelles for smart drug delivery systems.

Facile In Situ Preparation and In Vitro Antibacterial Activity of PDMAEMA-Based Silver-Bearing Copolymer Micelles.

Well-defined polymer micelles with core-shell structure are good delivery platform for stabilizing silver nanoparticles (AgNPs) in the field of antimicrobials targeting diseases. The rational construction of the polymer structure, an efficient, facile, and green preparation approach, and comprehensive exploration of the derived AgNPs are necessary, such as size, particle stability, antibacterial activity, and other properties. Herein, we designed and assessed the in vitro antimicrobial activity of AgNPs-decorated copolymer micelles with different copolymer topologies. First, linear or four-arm star triblock copolymers with the similar molecular weight and degree of polymerization were obtained, which consisted of DMAEMA for in situ reduction of silver ions to form AgNPs without external reducing agent. HEMA and PEGMA in micellar shell gave an enhanced stability of AgNPs during blood circulation. The combination of computational modeling and experimental results indicated that both types of micelles could fabricate AgNPs with monodisperse and spherical morphology. Star copolymer micelles stabilized AgNPs had smaller average size, better stability, and higher antibacterial activity than those with linear structure, which may due to higher stability of micelles from star copolymers. Furthermore, the cytotoxicity evaluation test showed that the achieved linear or star copolymers micelles stabilized AgNPs had good biocompatibility. This work provides a facile and universal approach in the rational design of micelles stabilized AgNPs with suitable topology for fighting against a wide range of bacterial infections.

DPD studies on mixed micelles self-assembled from MPEG-PDEAEMA and MPEG-PCL for controlled doxorubicin release.

In order to better understand and improve the drug loading capacity and release behavior of the pH-responsive mixed micelles in well controlled pH environments, dissipative particle dynamics (DPD) simulations are employed. This is performed by studying the co-micellization behavior of these materials produced from the two specific diblock polymers, poly(ethylene glycol) methyl ether-b-poly(N, N diethylamino ethyl methacrylate) (MPEG-PDEAEMA) and poly(ethylene glycol) methyl ether-b-polycaprolactone (MPEG-PCL) for doxorubicin (DOX) delivery. With the use of appropriate interaction parameters, the formation mechanism of (drug-loaded) mixed micelles, particle sizes, morphology, and composition are investigated. Simulation results show that compared with pure MPEG-PDEAEMA or MPEG-PCL, the mixed MPEG-PDEAEMA and MPEG-PCL system can combine to form multifunctional mixed micelles with larger particle sizes that lead to improved stability, higher drug loading capacity and better-controlled drug release performance. Simulations of the drug release process using the mixed micelles show that, when the environment is acidic, the tertiary amine group of PDEAEMA and DOX3 lead to rapid diffusion and release of the DOX in the aqueous solution. It is found that the presence of MPEG-PCL has a great influence in avoiding the fast release of the drug inside the core of micelles. Therefore, this study offers a deeper understanding of the mechanism on the co-micellization behaviors, the pH-responsive and drug controlled release behaviors of mixed micelles.

Controlled construction of gold nanoparticles in situ from ß-cyclodextrin based unimolecular micelles for in vitro computed tomography imaging.

The development of nanomaterials as highly efficient contrast agents for tumor computed tomography (CT) imaging still remains a huge challenge. In this study, a novel and facile approach to fabricate unimolecular micelles-stablized gold nanoparticles (AuNPs) without external reductant for in vitro targeted CT imaging was described. Amphiphilic 21-arm star-like polymers ß-cyclodextrin-g-{poly(2-(dimethylamino)ethyl methacrylate)-poly(2-hydroxyethyl methacrylate)-poly[poly(ethylene glycol) methyl ether methacrylate]} [ß-CD-g-(PDMA-b-PHEMA-b-PPEGMA)] was firstly synthesized and proved to form unimolecular core-middle layer-shell-type micelles in water through experimental and computer simulation results. Taking advantage of the reducing groups of PDMA block, AuNPs were decorated in the micellar PDMA block because of the in situ reduction of gold ions, which were absorbed by the PDMA chains in the core layer with a narrow nanoparticle size distribution. This strategy could prevent aggregation of AuNPs, which were capable of being employing as a highly effective probe for specific CT imaging in vitro. Importantly, the ß-CD-g-(PDMA-b-PHEMA-b-PPEGMA)/AuNPs incubated with HepG2 cells, displayed more intense X-ray attenuation property (>37%) than conventional iodine-based CT imaging agent (Omnipaque) and also possessed a satisfying cytocompatibility in the given concentration range. The facile fabrication procedures and the efficiency of CT imaging render the novel hybrid unimolecular micelles to become potent candidates for applications in tumor-targeted CT imaging.

Delivery of anticancer drug using pH-sensitive micelles from triblock copolymer MPEG-b-PBAE-b-PLA.

To improve the drug release rate in well-controlled manner, a new pH-sensitive triblock amphiphilic copolymer methyl poly(ethylene glycol) ether-b-poly(ß-amino esters)-b-poly lactic acid (MPEG-b-PBAE-b-PLA) and its self-assembled micelles were developed for anticancer drug delivery. The average molecular weight and molecular structure of MPEG-b-PBAE-b-PLA were confirmed by gel permeation chromatography (GPC) and 1H NMR. The formation of self-assembled micelles, the microstructures at different pH values, and the distribution of doxorubicin (DOX) were investigated by dissipative particle dynamics (DPD) simulation combined with experimental techniques. The copolymers formed stable core-shell-type micelles in water. The critical micelle concentration (CMC) values, particle sizes and zeta potentials of the blank micelles increased along with globule-extended conformational transitions when the pH values decreased from 7.4 to 5.0, due to the protonation of amine groups of PBAE. Obvious increases in the particle sizes and the drug loading content of micelles were observed with increasing DOX. The in vitro release behavior of DOX from the micelles was pH-dependent. The DOX release rate was improved obviously as pH decreased from pH7.4 to pH5.0, with over 96% of DOX was released within 48h. The drug release mechanism under different conditions was also analyzed using theoretical formulas. All the results suggest that the pH-sensitive MPEG-b-PBAE-b-PLA micelles might be a prospective candidate as anticancer drug delivery carrier with well-controlled release behavior.

pH-Sensitive Micelles Based on Star Copolymer Ad-(PCL-b-PDEAEMA-b-PPEGMA)4 for Controlled Drug Delivery.

Enhancing drug loading efficacy and stability of polymeric micelles remains a grand challenge. Here we develop adamantane-based star copolymers adamantane-[poly(ε-caprolactone)-b-poly(2-(diethylamino)ethyl methacrylate)-b-poly(poly(ethylene glycol) methyl ether methacrylate)]4 (Ad-(PCL-b-PDEAEMA-b-PPEGMA)4) and their self-assembled micelles for controlled drug delivery. Results show that the polymers have excellent stability in solution with low critical micelle concentration (CMC) (0.0025⁻0.0034 mg/mL) and the apparent base dissociation constant (pKb) of the polymers is from 5.31 to 6.05. Dynamic light scattering analysis exhibits the great environmental response capability of the pH-sensitive micelles according to particle sizes and zeta potentials. With the synergy effect of the adamantane and hydrophobic block, the micelles display the high Doxorubicin (DOX) loading efficacy (up to 22.4%). The DOX release study shows that the micelles are capable of controlled release for drug. This work indicates the Ad-(PCL-b-PDEAEMA-b-PPEGMA)4 micelles may provide new guidelines for drug control and release system in overcoming cancer treatment.

[Research and development of Lipin family.].

Lipin family including at least three members Lipin 1, Lipin 2, and Lipin 3 is a critical regulatory enzyme identified recently, which plays dual roles in lipid metabolisms. Lipin family has physiological effects not only on regulating lipid metabolism, but also on maintaining normal peripheral nervous functions, liver lipoprotein secretion, cell morphous, reproductive functions, and energy homeostasis. Since mutations in Lipin gene express may be associated with AIDS, insulin resistance, obesity, diabetes mellitus, and the other diseases of metabolic syndrome, Lipin may be a new useful target in treatment of above-mentioned clinical-related diseases. In this article, we focused on discovery, construction features, expression, regulatory mechanism, and biological functions of Lipin, as well as its correlation research with clinical-related diseases.