Natural biomacromolecules have attracted increased attention as carriers in biomedicine in recent years because of their inherent biochemical and biophysical properties including renewability,nontoxicity, biocompatibi...Natural biomacromolecules have attracted increased attention as carriers in biomedicine in recent years because of their inherent biochemical and biophysical properties including renewability,nontoxicity, biocompatibility, biodegradability, long blood circulation time and targeting ability. Recent advances in our understanding of the biological functions of natural-origin biomacromolecules and the progress in the study of biological drug carriers indicate that such carriers may have advantages over synthetic material-based carriers in terms of half-life, stability, safety and ease of manufacture. In this review, we give a brief introduction to the biochemical properties of the widely used biomacromoleculebased carriers such as albumin, lipoproteins and polysaccharides. Then examples from the clinic and in recent laboratory development are summarized. Finally the current challenges and future prospects of present biological carriers are discussed.展开更多
Protein therap34 wherein therapeutic proteins are delivered to treat disorders, is considered the safest and most direct approach for treating diseases. However, its applications are highly limited by the paucity of e...Protein therap34 wherein therapeutic proteins are delivered to treat disorders, is considered the safest and most direct approach for treating diseases. However, its applications are highly limited by the paucity of efficient strategies for delivering proteins and the rapid clearance of therapeutic proteins in vivo after their administration. Here, we demonstrate a novel strategy that can significantly prolong the circulation time of therapeutic proteins as well as minimize their immunogenicity. This is achieved by encapsulating individual protein molecules with a thin layer of crosslinked phosphorylcholine polymer that resists protein adsorption. Through extensive cellular studies, we demonstrate that the crosslinked phosphorylcholine polymer shell effectively prevents the encapsulated protein from being phagocytosed by macrophages, which play an essential role in the clearance of nanoparfides in vivo. Moreover, the polymer shell prevents the encapsulated protein from being identified by immune cells. As a result, immune responses against the therapeutic protein are effectively suppressed. This work describes a feasible method to prolong the circulation time and reduce the immunogenicity of therapeutic proteins, which may promote the development and application of novel protein therapies in the treatment of diverse diseases.展开更多
Therapeutic proteins and peptides have revolutionized treatment for a number of diseases, and the expected increase in macromolecule-based therapies brings a new set of challenges for the pharmaceutics field. Due to t...Therapeutic proteins and peptides have revolutionized treatment for a number of diseases, and the expected increase in macromolecule-based therapies brings a new set of challenges for the pharmaceutics field. Due to their poor stability, large molecular weight, and poor transport properties,therapeutic proteins and peptides are predominantly limited to parenteral administration. The short serum half-lives typically require frequent injections to maintain an effective dose, and patient compliance is a growing issue as therapeutic protein treatments become more widely available. A number of studies have underscored the relationship of subcutaneous injections with patient non-adherence, estimating that over half of insulin-dependent adults intentionally skip injections. The development of oral formulations has the potential to address some issues associated with non-adherence including the interference with daily activities, embarrassment, and injection pain. Oral delivery can also help to eliminate the adverse effects and scar tissue buildup associated with repeated injections. However, there are several major challenges associated with oral delivery of proteins and peptides, such as the instability in the gastrointestinal(GI)tract, low permeability, and a narrow absorption window in the intestine. This review provides a detailed overview of the oral delivery route and associated challenges. Recent advances in formulation and drugdelivery technologies to enhance bioavailability are discussed, including the co-administration of compounds to alter conditions in the GI tract, the modification of the macromolecule physicochemical properties, and the use of improved targeted and controlled release carriers.展开更多
Lipid nanoparticle(LNP)-based drug delivery systems have become the most clinically advanced non-viral delivery technology.LNPs can encapsulate and deliver a wide variety of bioactive agents,including the small molecu...Lipid nanoparticle(LNP)-based drug delivery systems have become the most clinically advanced non-viral delivery technology.LNPs can encapsulate and deliver a wide variety of bioactive agents,including the small molecule drugs,proteins and peptides,and nucleic acids.However,as the physicochemical properties of small-and macromolecular cargos can vary drastically,every LNP carrier system needs to be carefully tailored in order to deliver the cargo molecules in a safe and efficient manner.Our group applied the combinatorial library synthesis approach and in vitro and in vivo screening strategy for the development of LNP delivery systems for drug delivery.In this Review,we highlight our recent progress in the design,synthesis,characterization,evaluation,and optimization of combinatorial LNPs with novel structures and properties for the delivery of small-and macromolecular therapeutics both in vitro and in vivo.These delivery systems have enormous potentials for cancer therapy,antimicrobial applications,gene silencing,genome editing,and more.We also discuss the key challenges to the mechanistic study and clinical translation of new LNP-enabled therapeutics.展开更多
High MW chitosan (CS) solutions have already been proposed as vehicles for protein delivery. The aim of the present work is to investigate the potential utility of water-soluble chitosan (WSC) as vehicles to load and ...High MW chitosan (CS) solutions have already been proposed as vehicles for protein delivery. The aim of the present work is to investigate the potential utility of water-soluble chitosan (WSC) as vehicles to load and deliver proteins. WSC nanoparticles (WSC NP) with various formations were prepared based on ionic gelation of WSC with pentasodium tripolyphosphate (TPP) anions. Bovine serum albumin (BSA) was used as a model protein drug incorporated into the WSC nanoparticles. Blank and BSA-loaded WSC nanoparticles were examined and determined to have a spherical shape with diameters between 35―190 nm, and zeta potential between 35―42 mV. FTIR confirmed that the tripolyphosphoric groups of TPP linked to the ammonium groups of WSC in the nanoparticles. Some factors affecting delivery properties of BSA have been investigated. Altering the concentration of BSA from 0.05 to 1 mg/mL enhanced the loading capacity of BSA but decreased loading efficiency simultaneously. Also, with the introduction of poly ethylene glycol (PEG), BSA release accelerated. Nanoparticle preparation from WSC with various deacetylation degrees (DDs) from 72.6% to 90% and MWs ranging from 3.5 to 15.8 kDa promoted loading efficiency and decreased the release rate. These results indicate that WSC nanoparticles are promising carriers for protein delivery.展开更多
病毒样颗粒(virus like particles,VLPs)近年来被广泛应用于疫苗和治疗性药物递送系统领域。与体内组装VLPs相比,体外组装VLPs具有组装寡聚体成分明确、反应条件可控、避免宿主成分的引入以及能实现药物活性成分的包裹和递送功能等诸多...病毒样颗粒(virus like particles,VLPs)近年来被广泛应用于疫苗和治疗性药物递送系统领域。与体内组装VLPs相比,体外组装VLPs具有组装寡聚体成分明确、反应条件可控、避免宿主成分的引入以及能实现药物活性成分的包裹和递送功能等诸多优势。概述了体外VLPs的生产方法、影响体外VLPs形成的因素、VLPs的特征分析方法,综述了体外VLPs在疫苗和治疗性药物递送方面的应用进展,期望对体外VLPs组装技术的发展提供参考思路,促进体外VLPs组装技术在疾病预防和治疗应用方面发挥更大的作用。展开更多
Controlled delivery of proteins and other biologics is a growing medium of therapy for diseases previously untreatable.Here we report a self-assembling,tunable vesicle for the controlled delivery of growth factors and...Controlled delivery of proteins and other biologics is a growing medium of therapy for diseases previously untreatable.Here we report a self-assembling,tunable vesicle for the controlled delivery of growth factors and cytokines.Coacervate made of heparin and a biocompatible polycation,PEAD,forms the core of the vesicle;lipids form the membrane of the vesicle.We call this vesicle lipocoacervate(LipCo),which has a high affinity for growth factors and cytokines due to heparin.LipCo is a tunable protein delivery vehicle.The vesicle size is controlled through polymer and salt concentrations.Membrane functionalization enables potential for targeting capabilities with long-term storage through lyophilization.Importantly,the controlled delivery of therapeutics also avoids high toxicity to treated cells in vitro.Here we report on these key principles of LipCo assembly and design.展开更多
Polymer conjugation was found highly valuable in clinic to improve the bioavailability of protein therapeutics.However,it is still a tremendous challenge to achieve a complete release of original proteins from the con...Polymer conjugation was found highly valuable in clinic to improve the bioavailability of protein therapeutics.However,it is still a tremendous challenge to achieve a complete release of original proteins from the conjugated hybrid under external stimulus to recover active proteins in the targeted tissue.Herein,we report a general light-controlled protein delivery methodology by weaving a photodegradable polymer cocoon around proteins,which could reliably protect them from degradation in the dark while efficiently releasing them under illumination without any residual atoms.The surface charge of the polymer shell is easily tunable to facilitate efficient cell uptake.The versatility of this strategy is demonstrated by the delivery of the Cas9/sg RNA complex that realized light-controlled gene editing both in vitro and in vivo,and such repertoire is of particular value in regard to minimizing the off-target toxicity of CRISPR-Cas9-based gene therapy.展开更多
Lacking a precise targeting strategy,castration-resistant prostate cancer(CRPC)is still hard to be treat effectively.Exploring treatment options that can accurately target CPRC is an important issue with urgent need.I...Lacking a precise targeting strategy,castration-resistant prostate cancer(CRPC)is still hard to be treat effectively.Exploring treatment options that can accurately target CPRC is an important issue with urgent need.In this study,a novel nanotechnologybased strategy had been developed for the precise target treatment of CRPC.By combining microwaves and photothermal therapy(PTT),this nanoplatform,cmHSP70-PL-AuNC-DOX,targets tumor tissues with outstanding precision and achieves better anti-tumor activity by simultaneously eliciting photothermal and chemotherapeutic effects.From nanotechnology,cmHSP70-modified and thermo-sensitive liposome-coated AuNC-DOX were prepared and used for CRPC-targeted photothermal ablation and chemotherapy.Doxorubicin(DOX)was selected as the chemotherapeutic agent for cytotoxicity.In terms of the curative scheme,prostate tissues were firstly pre-treated with microwaves to induce the expression of heat shock protein 70(HSP70)and its migration to the cell membrane,which was then targeted by HSP70 antibody(cmHSP70)coated on the nanoparticles to achieve accurate drug delivery.The nanoplatform then achieved precise ablation and controlled release of DOX under external near-infrared(NIR)irradiation.Through the implementation,the targeting,cell killing,and safety of this therapeutical strategy had been verified in vivo and in vitro.This work establishes an accurate,controllable,efficient,non-invasive,and safe treatment platform for targeting CRPC,provides a rational design for CRPC’s PTT,and offers new prospects for nanomedicines with great precision.展开更多
Intracellular protein delivery is critical to the development of protein-based biopharmaceuticals and therapies.However,current delivery vectors often suffer from complicated syntheses,low generality among various pro...Intracellular protein delivery is critical to the development of protein-based biopharmaceuticals and therapies.However,current delivery vectors often suffer from complicated syntheses,low generality among various proteins,and insufficient serum stability.Herein,we developed an enlightened cytosolic protein delivery strategy by dynamically crosslinking epigallocatechin gallate(EGCG),low-molecular-weight polyethylenimine(PEI 1.8k),and 2-acetylphenylboric acid(2-APBA)on the protein surface,hence forming the EPP-protein nanocapsules(NCs).EGCG enhanced protein encapsulation via hydrogen bonding,and reduced the positive charge density of PEI to endow the NCs with high serum tolerance,thereby enabling effective cellular internalization in serum.The formation of reversible imine and boronate ester among 2-APBA,EGCG,and PEI 1.8k allowed acid-triggered dissociation of EPP-protein NCs in the endolysosomes,which triggered efficient intracellular release of the native proteins.Such strategy therefore showed high efficiency and universality for diversities of proteins with different molecular weights and isoelectric points,including enzyme,toxin,antibody,and CRISPR(clustered regularly interspaced short palindromic repeats)-Cas9 ribonucleoprotein(RNP),outperforming the commercial protein transduction reagent PULSin and RNP transfection reagent lipofectamine CMAX.Moreover,intravenously(i.v.)injected EPP-saporin NCs efficiently delivered saporin into 4T1 tumor cells to provoke robust antitumor effect.This simple,versatile,and robust cytosolic protein delivery system holds translational potentials for the development of protein-based therapeutics.展开更多
文摘Natural biomacromolecules have attracted increased attention as carriers in biomedicine in recent years because of their inherent biochemical and biophysical properties including renewability,nontoxicity, biocompatibility, biodegradability, long blood circulation time and targeting ability. Recent advances in our understanding of the biological functions of natural-origin biomacromolecules and the progress in the study of biological drug carriers indicate that such carriers may have advantages over synthetic material-based carriers in terms of half-life, stability, safety and ease of manufacture. In this review, we give a brief introduction to the biochemical properties of the widely used biomacromoleculebased carriers such as albumin, lipoproteins and polysaccharides. Then examples from the clinic and in recent laboratory development are summarized. Finally the current challenges and future prospects of present biological carriers are discussed.
基金This work is supported by the National Natural Science Foundation of China （NSFC, Nos. 91127045, 51390483, 51473319, 51303025, 81401439 and 51343007）, YG2012MS38 and China Postdoctoral Science Foundation （No. 2014M551399）.
文摘Protein therap34 wherein therapeutic proteins are delivered to treat disorders, is considered the safest and most direct approach for treating diseases. However, its applications are highly limited by the paucity of efficient strategies for delivering proteins and the rapid clearance of therapeutic proteins in vivo after their administration. Here, we demonstrate a novel strategy that can significantly prolong the circulation time of therapeutic proteins as well as minimize their immunogenicity. This is achieved by encapsulating individual protein molecules with a thin layer of crosslinked phosphorylcholine polymer that resists protein adsorption. Through extensive cellular studies, we demonstrate that the crosslinked phosphorylcholine polymer shell effectively prevents the encapsulated protein from being phagocytosed by macrophages, which play an essential role in the clearance of nanoparfides in vivo. Moreover, the polymer shell prevents the encapsulated protein from being identified by immune cells. As a result, immune responses against the therapeutic protein are effectively suppressed. This work describes a feasible method to prolong the circulation time and reduce the immunogenicity of therapeutic proteins, which may promote the development and application of novel protein therapies in the treatment of diverse diseases.
基金supported in part by a grant from the National Institutes of Health (R01-EB-00246020)the Cockrell Family Regents Chair. Angela M.Wagner was supported by a National Science Foundation Graduate Research Fellowship (DGE-1610403)+1 种基金the S.E.S.H.A. Endowed Graduate Fellowship in Engineeringthe Philanthropic Educational Organization Scholar Award
文摘Therapeutic proteins and peptides have revolutionized treatment for a number of diseases, and the expected increase in macromolecule-based therapies brings a new set of challenges for the pharmaceutics field. Due to their poor stability, large molecular weight, and poor transport properties,therapeutic proteins and peptides are predominantly limited to parenteral administration. The short serum half-lives typically require frequent injections to maintain an effective dose, and patient compliance is a growing issue as therapeutic protein treatments become more widely available. A number of studies have underscored the relationship of subcutaneous injections with patient non-adherence, estimating that over half of insulin-dependent adults intentionally skip injections. The development of oral formulations has the potential to address some issues associated with non-adherence including the interference with daily activities, embarrassment, and injection pain. Oral delivery can also help to eliminate the adverse effects and scar tissue buildup associated with repeated injections. However, there are several major challenges associated with oral delivery of proteins and peptides, such as the instability in the gastrointestinal(GI)tract, low permeability, and a narrow absorption window in the intestine. This review provides a detailed overview of the oral delivery route and associated challenges. Recent advances in formulation and drugdelivery technologies to enhance bioavailability are discussed, including the co-administration of compounds to alter conditions in the GI tract, the modification of the macromolecule physicochemical properties, and the use of improved targeted and controlled release carriers.
基金supported by the National Institutes of Health(NIH)Grants R01 EB027170-04 and UG3 TR002636-01,USA。
文摘Lipid nanoparticle(LNP)-based drug delivery systems have become the most clinically advanced non-viral delivery technology.LNPs can encapsulate and deliver a wide variety of bioactive agents,including the small molecule drugs,proteins and peptides,and nucleic acids.However,as the physicochemical properties of small-and macromolecular cargos can vary drastically,every LNP carrier system needs to be carefully tailored in order to deliver the cargo molecules in a safe and efficient manner.Our group applied the combinatorial library synthesis approach and in vitro and in vivo screening strategy for the development of LNP delivery systems for drug delivery.In this Review,we highlight our recent progress in the design,synthesis,characterization,evaluation,and optimization of combinatorial LNPs with novel structures and properties for the delivery of small-and macromolecular therapeutics both in vitro and in vivo.These delivery systems have enormous potentials for cancer therapy,antimicrobial applications,gene silencing,genome editing,and more.We also discuss the key challenges to the mechanistic study and clinical translation of new LNP-enabled therapeutics.
文摘High MW chitosan (CS) solutions have already been proposed as vehicles for protein delivery. The aim of the present work is to investigate the potential utility of water-soluble chitosan (WSC) as vehicles to load and deliver proteins. WSC nanoparticles (WSC NP) with various formations were prepared based on ionic gelation of WSC with pentasodium tripolyphosphate (TPP) anions. Bovine serum albumin (BSA) was used as a model protein drug incorporated into the WSC nanoparticles. Blank and BSA-loaded WSC nanoparticles were examined and determined to have a spherical shape with diameters between 35―190 nm, and zeta potential between 35―42 mV. FTIR confirmed that the tripolyphosphoric groups of TPP linked to the ammonium groups of WSC in the nanoparticles. Some factors affecting delivery properties of BSA have been investigated. Altering the concentration of BSA from 0.05 to 1 mg/mL enhanced the loading capacity of BSA but decreased loading efficiency simultaneously. Also, with the introduction of poly ethylene glycol (PEG), BSA release accelerated. Nanoparticle preparation from WSC with various deacetylation degrees (DDs) from 72.6% to 90% and MWs ranging from 3.5 to 15.8 kDa promoted loading efficiency and decreased the release rate. These results indicate that WSC nanoparticles are promising carriers for protein delivery.
文摘病毒样颗粒(virus like particles,VLPs)近年来被广泛应用于疫苗和治疗性药物递送系统领域。与体内组装VLPs相比,体外组装VLPs具有组装寡聚体成分明确、反应条件可控、避免宿主成分的引入以及能实现药物活性成分的包裹和递送功能等诸多优势。概述了体外VLPs的生产方法、影响体外VLPs形成的因素、VLPs的特征分析方法,综述了体外VLPs在疫苗和治疗性药物递送方面的应用进展,期望对体外VLPs组装技术的发展提供参考思路,促进体外VLPs组装技术在疾病预防和治疗应用方面发挥更大的作用。
文摘Controlled delivery of proteins and other biologics is a growing medium of therapy for diseases previously untreatable.Here we report a self-assembling,tunable vesicle for the controlled delivery of growth factors and cytokines.Coacervate made of heparin and a biocompatible polycation,PEAD,forms the core of the vesicle;lipids form the membrane of the vesicle.We call this vesicle lipocoacervate(LipCo),which has a high affinity for growth factors and cytokines due to heparin.LipCo is a tunable protein delivery vehicle.The vesicle size is controlled through polymer and salt concentrations.Membrane functionalization enables potential for targeting capabilities with long-term storage through lyophilization.Importantly,the controlled delivery of therapeutics also avoids high toxicity to treated cells in vitro.Here we report on these key principles of LipCo assembly and design.
基金supported by the National Key R&D Program of China (2018YFA0903500)the Max-Planck Partner Group+1 种基金the National Natural Science Foundation of China (22077042)the National Key Research and Development Program of China (2022YFA1206001 to S.G.)。
文摘Polymer conjugation was found highly valuable in clinic to improve the bioavailability of protein therapeutics.However,it is still a tremendous challenge to achieve a complete release of original proteins from the conjugated hybrid under external stimulus to recover active proteins in the targeted tissue.Herein,we report a general light-controlled protein delivery methodology by weaving a photodegradable polymer cocoon around proteins,which could reliably protect them from degradation in the dark while efficiently releasing them under illumination without any residual atoms.The surface charge of the polymer shell is easily tunable to facilitate efficient cell uptake.The versatility of this strategy is demonstrated by the delivery of the Cas9/sg RNA complex that realized light-controlled gene editing both in vitro and in vivo,and such repertoire is of particular value in regard to minimizing the off-target toxicity of CRISPR-Cas9-based gene therapy.
基金This study was supported by the National Natural Science Foundation of China(Nos.82172679 and 82104405)Zhejiang Provincial Medicine and Health Science Foundation(No:2021KY010).
文摘Lacking a precise targeting strategy,castration-resistant prostate cancer(CRPC)is still hard to be treat effectively.Exploring treatment options that can accurately target CPRC is an important issue with urgent need.In this study,a novel nanotechnologybased strategy had been developed for the precise target treatment of CRPC.By combining microwaves and photothermal therapy(PTT),this nanoplatform,cmHSP70-PL-AuNC-DOX,targets tumor tissues with outstanding precision and achieves better anti-tumor activity by simultaneously eliciting photothermal and chemotherapeutic effects.From nanotechnology,cmHSP70-modified and thermo-sensitive liposome-coated AuNC-DOX were prepared and used for CRPC-targeted photothermal ablation and chemotherapy.Doxorubicin(DOX)was selected as the chemotherapeutic agent for cytotoxicity.In terms of the curative scheme,prostate tissues were firstly pre-treated with microwaves to induce the expression of heat shock protein 70(HSP70)and its migration to the cell membrane,which was then targeted by HSP70 antibody(cmHSP70)coated on the nanoparticles to achieve accurate drug delivery.The nanoplatform then achieved precise ablation and controlled release of DOX under external near-infrared(NIR)irradiation.Through the implementation,the targeting,cell killing,and safety of this therapeutical strategy had been verified in vivo and in vitro.This work establishes an accurate,controllable,efficient,non-invasive,and safe treatment platform for targeting CRPC,provides a rational design for CRPC’s PTT,and offers new prospects for nanomedicines with great precision.
基金supported by the Natural Science Foundation of Jiangsu Province(No.BK20220245)the National Natural Science Foundation of China(Nos.52273144 and 82241008)Collaborative Innovation Center of Suzhou Nano Science&Technology,the 111 project,Suzhou Key Laboratory of Nanotechnology and Biomedicine,and Joint International Research Laboratory of Carbon-Based Functional Materials and Devices.
文摘Intracellular protein delivery is critical to the development of protein-based biopharmaceuticals and therapies.However,current delivery vectors often suffer from complicated syntheses,low generality among various proteins,and insufficient serum stability.Herein,we developed an enlightened cytosolic protein delivery strategy by dynamically crosslinking epigallocatechin gallate(EGCG),low-molecular-weight polyethylenimine(PEI 1.8k),and 2-acetylphenylboric acid(2-APBA)on the protein surface,hence forming the EPP-protein nanocapsules(NCs).EGCG enhanced protein encapsulation via hydrogen bonding,and reduced the positive charge density of PEI to endow the NCs with high serum tolerance,thereby enabling effective cellular internalization in serum.The formation of reversible imine and boronate ester among 2-APBA,EGCG,and PEI 1.8k allowed acid-triggered dissociation of EPP-protein NCs in the endolysosomes,which triggered efficient intracellular release of the native proteins.Such strategy therefore showed high efficiency and universality for diversities of proteins with different molecular weights and isoelectric points,including enzyme,toxin,antibody,and CRISPR(clustered regularly interspaced short palindromic repeats)-Cas9 ribonucleoprotein(RNP),outperforming the commercial protein transduction reagent PULSin and RNP transfection reagent lipofectamine CMAX.Moreover,intravenously(i.v.)injected EPP-saporin NCs efficiently delivered saporin into 4T1 tumor cells to provoke robust antitumor effect.This simple,versatile,and robust cytosolic protein delivery system holds translational potentials for the development of protein-based therapeutics.
