CHEMICAL ENGINEERING JOURNAL
·
2025
·
IF 13.2
Liqun Xia,Yi Chen,Haiyang Wu,Mingchao Wang,Gonghui Li
Reaction engineering is key to enhance enzyme-catalyzed reactions. While previous studies have focused on boosting enzyme activity, they often overlook enzyme kinetics—especially the K m ( S 0.5 ) value—that are crucial for optimal performance under physiological conditions. Herein, we developed a novel enzyme-kinetics-guided reaction engineering strategy to improve the substrate affinity of therapeutic enzymes. In this proof-of-concept experiment, we selected arginine deiminase (ADI), a promising therapeutic enzyme for treating arginine-auxotrophic tumors. First, a kinetic trap was identified, and an enzyme-kinetics-based screening model was developed to overcome it. Next, we identified and characterized a novel arginine deiminase (ADI) from Gammaproteobacteria ( Gam ADI), which was subsequently engineered to enhance its catalytic efficiency under the guidance of the kinetic model. Through ultra-high-throughput screening, two ADI variants with low S 0.5 values were identified. After recombination, the final variant GamADI M7 exhibited a remarkable improvement in substrate affinity (91 % reduction in S 0.5 value) and a 1382-fold enhancement in catalytic efficiency. The therapeutic potential of GamADI M7 was validated via cytotoxicity assays in melanoma and prostate cancer cell lines, where its IC 50 values decreased dramatically compared to wild-type ADI. This study highlights the power of enzyme-kinetics-guided reaction engineering, offering a promising strategy to enhance enzyme substrate affinity.
研究领域:
生物化学
·
分子生物学
·
肿瘤学
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产品关联
DOI
高影响因子
ACS Applied Materials & Interfaces
·
2020
·
IF 8.758
Daxiu Li,Xia Li,Fang Yang,Ruo Yuan,Yun Xiang
A protein-based therapy is significantly challenged by the successful delivery of native proteins into the targeted cancer cells. We address this challenge here using an all-sealed divalent aptamer tetrahedral DNA framework (asdTDF) delivery platform, in which the protein drug is encapsulated inside the cavity of the framework stoichiometrically via a reversible chemical bond. The ligase-assisted seal of the nicks results in highly enhanced TDF stability of the against nuclease digestion to effectively protect the therapeutic protein from degradation. In addition, the divalent aptamer sequences incorporated into the framework favor it with a target-specific and efficient delivery capability. Importantly, upon being readily delivered into the targeted cancer cells, endogenous glutathione can trigger the release of the native therapeutic protein from the TDF in a traceless fashion by cleaving the reversible chemical bond, thereby leading to effective apoptosis of the specific cancer cells.
研究领域:
生物技术
·
医学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
BIOMACROMOLECULES
·
2016
·
IF 5.583
Xing Zhou,Yang Zhao,Siyu Chen,Songling Han,Xiaoqiu Xu,Jiawei Guo,Mengyu Liu,Ling Che,Xiaohui Li,Jianxiang Zhang
Targeted delivery of therapeutics to the intestine is preferred for the management of many diseases due to its diverse advantages. Currently, there are still challenges in creating cost-effective and translational pH-responsive microspheres for intestinal delivery of various hydrophobic drugs. Herein we report a multiple noncovalent interactions-mediated assembly strategy in which carboxyl-bearing compounds (CBCs) are guest molecules, while poly(N-isopropylacrylamide) (PNIPAm) serves as a host polymer. Formation of microparticles and therapeutic packaging can be achieved simultaneously by this assembly approach, leading to well-shaped microspheres with extremely higher drug loading capacity as compared to microspheres based on two FDA-approved materials of poly(d,l-lactide-co-glycolide) (PLGA) and an enteric coating polymer EudragitS 100 (S100). Also, carboxyl-deficient hydrophobic drugs can be effectively entrapped. These assembled microspheres, with excellent reconstitution capability as well as desirable scalability, could selectively release drug molecules under intestinal conditions. By significantly enhancing drug dissolution/release in the intestine, these pH-responsive assemblies may notably improve the oral bioavailability of loaded therapeutics. Moreover, the assembled microspheres possessed superior therapeutic performance in rodent models of inflammation and tumor over the control microspheres derived from PLGA and S100. Therapy with newly developed microspheres did not cause undesirable side effects. Furthermore, in vivo evaluation in mice revealed the carrier material PNIPAm was safe for oral delivery at doses as high as 10 g/kg. Collectively, our findings demonstrated that this type of pH-responsive microsphere may function as superior and translational intestine-directed delivery systems for a diverse array of therapeutics.
研究领域:
药剂学
·
生物医学工程
·
药物递送系统
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产品关联
DOI
PubMed
高影响因子
Nanomaterials
·
2022
·Vol. 12
·期 4
·625
·
IF 5.719
Chao Deng,Xianghai Li,Qianru Jin,Deliang Yi
Enzyme therapy has important implications for the treatment of metabolic disorders and biological detoxification. It remains challenging to prepare enzymatic nanoreactors with high therapeutic efficiency and low emission of cytotoxic reaction intermediates. Here, we propose a novel strategy for the preparation of enzymes-loaded polypeptide microcapsules (EPM) with concentrically encapsulated enzymes to achieve higher cascade reaction rates and minimal emission of cytotoxic intermediates. Mesoporous silica spheres (MSS) are used as a highly porous matrix to efficiently load a therapeutic enzyme (glucose oxidase, GOx), and a layer-by-layer (LbL) assembly strategy is employed to assemble the scavenging enzyme (catalase) and polyelectrolyte multilayers on the MSS surface. After removal of the MSS, a concentrically encapsulated EPM is obtained with the therapeutic enzyme encapsulated inside the capsule, and the scavenging enzyme immobilized in the polypeptide multilayer shell. Performance of the concentrically encapsulated GOx-catalase capsules is investigated for synergistic glucose metabolism disturbance correction and cytotoxic intermediate H2O2clearance. The results show that the EPM can simultaneously achieve 99% H2O2clearance and doubled glucose consumption rate. This strategy can be extended to the preparation of other dual- or multi-enzyme therapeutic nanoreactors, showing great promise in the treatment of metabolic disorders.Keywords:therapeutic nanoreactor;mesoporous silica;enzyme therapy;capsule;metabolic disorder redressing
研究领域:
生物医学工程
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生物化学
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纳米医学
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产品关联
DOI
PubMed
高影响因子
Advanced Healthcare Materials
·
2018
·Vol. 8
·期 6
·1801010
·
IF 5.609
Zhipeng Zeng,Cong Dong,Pengfei Zhao,Zhijia Liu,Lixin Liu,Hai-Quan Mao,Kam W. Leong,Xiaohu Gao,Yongming Chen
Flash nanoprecipitation (FNP) by fast mixing of drug-containing organic solvent and water in a microchamber is a powerful and scalable technology to produce solid drug nanoparticles with high payload. The embedded therapeutic drugs, however, are largely limited to hydrophobic small molecules. By transferring proteins into organic solvent via hydrophobic ion pairing, the scope of FNP applications is expanded. This platform technology is capable of producing protein nanoparticles with tunable sizes (from ≈30 nm to sub-micrometers), high-production scale (grams per hour), high drug loading efficiency (>90%), and excellent reproducibility, opening a new paradigm for formulation of biological pharmaceuticals. As a proof-of-concept, insulin nanoparticles are made to address a major medical challenge; oral administration. A relative insulin bioavailability of 13.2% is achieved, enabling sustained reduction of blood glucose levels in a diabetic rat model.
研究领域:
药物递送
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生物技术
·
纳米医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Nature Communications
·
2022
·Vol. 13
·期 1
·1-11
·
IF 17.694
Pan Jiezhou,Gong Guidong,Wang Qin,Shang Jiaojiao,He Yunxiang,Catania Chelsea,Birnbaum Dan,Li Yifei,Jia Zhijun,Zhang Yaoyao,Joshi Neel S.,Guo Junling
The gut microbiota represents a large community of microorganisms that play an important role in immune regulation and maintenance of homeostasis. Living bacteria receive increasing interest as potential therapeutics for gut disorders, because they inhibit the colonization of pathogens and positively regulate the composition of bacteria in gut. However, these treatments are often accompanied by antibiotic administration targeting pathogens. In these cases, the efficacy of therapeutic bacteria is compromised by their susceptibility to antibiotics. Here, we demonstrate that a single-cell coating composed of tannic acids and ferric ions, referred to as ‘nanoarmor’, can protect bacteria from the action of antibiotics. The nanoarmor protects both Gram-positive and Gram-negative bacteria against six clinically relevant antibiotics. The multiple interactions between the nanoarmor and antibiotic molecules allow the antibiotics to be effectively absorbed onto the nanoarmor. Armored probiotics have shown the ability to colonize inside the gastrointestinal tracts of levofloxacin-treated rats, which significantly reduced antibiotic-associated diarrhea (AAD) resulting from the levofloxacin-treatment and improved some of the pre-inflammatory symptoms caused by AAD. This nanoarmor strategy represents a robust platform to enhance the potency of therapeutic bacteria in the gastrointestinal tracts of patients receiving antibiotics and to avoid the negative effects of antibiotics in the gastrointestinal tract.
研究领域:
微生物学
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药学
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纳米技术
·
胃肠病学
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DOI
PubMed
高影响因子
Biomaterials Science
·
2019
·Vol. 7
·期 7
·2934-2950
·
IF 5.251
Xinyi Tan,Yan Zhang,Qian Wang,Tianyang Ren,Jingxin Gou,Wei Guo,Tian Yin,Haibing He,Yu Zhang,Xing Tang
Poor permeation across intestinal mucous barriers often limits the oral delivery of prospective therapeutic proteins and peptides (TPPs). In order to address this issue, cell penetrating peptide (CPP) together with PEG modified and pore-enlarged mesostructured silica nanoparticle (NP) were constructed to form the mucus-penetrating electrostatic particle-complexes, CPP/TPP/NP. Alone, CPP and TPP often present with poor stability, and their traditional electrostatic complex shows reduced pharmacodynamics. To provide satisfactory protection, silica NPs were loaded with CPP and TPP (CPP@NP and TPP@NP), respectively, and then CPP@NP and TPP@NP could together form CPP/TPP/NP via electrostatic interaction. As a result, CPP involvement with PEG modification showed an 8.45-, 1.62- and 5.09-fold increase in cellular uptake, exocytosis and final transcellular permeation in mucous conditions, respectively. It was found that CPP involvement mainly affected transport and exocytosis, and the PEG polymer significantly influenced mucous penetration and cellular uptake, which could further promote CPP ability for uptake and exocytosis. Additionally, NP-mediated CPP/TPP/NP showed a similar uptake mechanism with supporting carriers (clathrin-mediated endocytosis), and could strengthen transcellular routes (the endoplasmic reticulum–Golgi apparatus pathway and the lysosome route). Utilizing recombinant growth hormone (RGH) as a model TPP, oral administration of the RGH-loaded CPP/TPP/LMSN-PEG10k with hydrophilic and electroneutral properties induced 5.41- and 4.91-fold increases in pharmacodynamics in vitro and in vivo, respectively. Thus, CPP/TPP/NP significantly promoted mucous permeation and shows promising potential for oral delivery of TPPs.
研究领域:
药学科学
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纳米技术
·
药物递送系统
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL IMMUNOPHARMACOLOGY
·
2022
·
IF 5.714
Jian-Bing Xiong,Jia-Xi Duan,Nan Jiang,Chen-Yu Zhang,Wen-Jing Zhong,Jin-Tong Yang,Yu-Biao Liu,Feng Su,Yong Zhou,Dai Li,Hui-Hui Yang,Cha-Xiang Guan
Our previous study showed that triggering receptors expressed on myeloid cell-1 (TREM-1) was upregulated in bleomycin (BLM)-induced pulmonary fibrosis (PF) mouse model. However, the role of TREM-1 in the development of PF and its underlying mechanism remain unclear. Herein, we report that the prophylactical blockade of TREM-1 using a decoy peptide dodecapeptide (LR12) exerted protective effects against BLM-induced PF in mice, with a higher survival rate, attenuated tissue injury, and less extracellular matrix deposition. Interestingly, therapeutic blockade of TREM-1 at the early stage of fibrosis also attenuated BLM-induced PF, suggesting a non-inflammatory effect. More importantly, we observed that TREM-1 blockade with LR12 significantly reduced the expression of the senescence-relative protein, including p16, p21, p53, and γ-H2AX in the lungs of PF mice. Notably, TREM-1 was upregulated in alveolar epithelial cells (AECs) and correlated with the levels of senescence markers in BLM-treated mice. In vitro , activating TREM-1 with an agonistic antibody exacerbated BLM-induced senescence in MLE12 cells, a murine AEC cell line. Furthermore, prophylactic or therapeutic blockade of TREM-1 protected MLE12 cells from senescence induced by BLM or H 2 O 2 . In conclusion, our findings elucidate a pro-fibrotic effect of TREM-1 by inducing AECs senescence in PF, providing a potential strategy for fibrotic disease treatment.
研究领域:
肺科学
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免疫学
·
细胞生物学
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药理学
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产品关联
DOI
PubMed
高影响因子
Small
·
2021
·Vol. 17
·期 45
·2102485
·
IF 13.281
Tianyou Wang,Qianqian Fan,Jiaxu Hong,Zhan Chen,Xujiao Zhou,Jianhua Zhang,Yiqin Dai,Hao Jiang,Zhipeng Gu,Yiyun Cheng,Yiwen Li
The therapeutic potential of nanomaterials toward oxidative damage relevant diseases has attracted great attentions by offering promising advantages compared with conventional antioxidants. Although different kinds of nanoantioxidants have been well developed, the facile fabrication of robust and efficient nanoscavengers is still met with challenges like the use of toxic and high-cost subunits, the involvement of multistep synthetic process, and redundant purification work. Herein, a direct fabrication strategy toward polyphenol nanoparticles with tunable size, excellent biocompatibility, and reactive oxygen species (ROS) scavenging capacities from grape seed via an enzymatic polymerization method is reported. The resulting nanoparticles can efficiently prevent cell damage from ROS and exert promising in vivo antioxidant therapeutic effects on several oxidative stress-related diseases, including accelerating wound healing, inhibiting ulcerative colitis, and regulating the oxidative stress in dry eye disease. This study can stimulate the development of more kinds of low-cost, safe, and efficient biomass-based antioxidative nanomaterials via similar fabrication methodologies.
研究领域:
纳米技术
·
药学
·
生物医学工程
·
材料科学
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产品关联
DOI
PubMed
高影响因子
Pharmaceutics
·
2024
·Vol. 16
·期 3
·422
·
IF 4.9
He Zhang,Anle Ge,Yulin Wang,Boran Xia,Xichu Wang,Zhonghui Zheng,Changsheng Wei,Bo Ma,Lin Zhu,Rose Amal,Sung Lai Jimmy Yun,Zi Gu
The therapeutic application of biofunctional proteins relies on their intracellular delivery, which is hindered by poor cellular uptake and transport from endosomes to cytoplasm. Herein, we constructed a two-dimensional (2D) ultrathin layered double hydroxide (LDH) nanosheet for the intracellular delivery of a cell-impermeable protein, gelonin, towards efficient and specific cancer treatment. The LDH nanosheet was synthesized via a facile method without using exfoliation agents and showed a high loading capacity of proteins (up to 182%). Using 2D and 3D 4T1 breast cancer cell models, LDH–gelonin demonstrated significantly higher cellular uptake efficiency, favorable endosome escape ability, and deep tumor penetration performance, leading to a higher anticancer efficiency, in comparison to free gelonin. This work provides a promising strategy and a generalized nanoplatform to efficiently deliver biofunctional proteins to unlock their therapeutic potential for cancer treatment.
研究领域:
纳米技术
·
药学
·
癌症研究
·
生物医学工程
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2014
·
IF 5.825
Lihua Zhang,Hua Liu,Yong Shao,Clement Lin,Huan Jia,Gang Chen,Danzhou Yang,Ying Wang
Aptamers, that exist naturally in living cells as functional elements and can switch nonfluorescent natural targets to fluorophores, are very useful in developing highly sensitive and selective biosensors and screening functional agents. This work demonstrates that human telomeric G-quadruplex (HTG) can serve as a potential fluorophore-switching aptamer (FSA) to target a natural isoquinoline alkaloid. We found that, among the G-quadruplexes studied here and the various structurally similar alkaloids including epiberberine (EPI), berberine (BER), palmatine (PAL), jatrorrhizine (JAT), coptisine (COP), worenine (WOR), sanguinarine (SAN), chelerythrine (CHE), and nitidine (NIT), only the HTG DNA, especially with a 5′-TA-3′ residue at the 5′ end of the G-quadruplex tetrad (5′-TAG3(TTAG3)3-3′, TA[Q]) as the minimal sequence, is the most efficient FSA to selectively light up the EPI fluorescence. Compared to the 5′ end flanking sequences, the 3′ end flanking sequences of the tetrad contribute significantly less to the recognition of EPI. The binding affinity of EPI to TA[Q] (Kd = 37 nM) is at least 20 times tighter than those of the other alkaloids. The steady-state absorption, steady-state/time-resolved fluorescence, and NMR studies demonstrate that EPI most likely interact with the 5′ end flanking sequence substructure beyond the core [Q] and the G-quadruplex tetrad in a much more specific manner than the other alkaloids. The highly selective and tight binding of EPI with the FSA and significantly enhanced fluorescence suggest the potential development of a selective EPI sensor (detection limit of 10 nM). More importantly, EPI, as the brightest FSA emitter among the alkaloids, can also serve as an efficient conformation probe for HTG DNA and discriminate the DNA G-quadruplex from the RNA counterpart. Furthermore, EPI can bind stoichiometrically to each G-quadruplex unit of long HTG DNA multimer with the most significant fluorescence enhancement, which has not been achieved by the previously reported probes. Our work suggests the potential use of EPI as a bioimaging probe and a therapeutic DNA binder.
研究领域:
分子生物学
·
化学生物学
·
纳米技术
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Frontiers in Pharmacology
·
2018
·
IF 3.831
Jiao Shihong,Tang Binghua,Wang Yong,Li Chun,Zeng Zifan,Cui Lixia,Zhang Xuefeng,Shao Mingyan,Guo Dongqing,Wang Qiyan
Coronary artery disease (CAD) is one of the leading causes of deaths worldwide. Energy metabolism disorders, including a reduction in fatty acids oxidation and upregulation of glycolysis pathway, are involved in the process of CAD. Therapeutic angiogenesis has become a promising treatment for CAD. Traditional Chinese medicines, such as Danqi Pill (DQP), have been proven to be effective in treating CAD in China for many years. However, the pro-angiogenic effects of DQP based on fatty acids oxidation are still unknown and the mechanism is worthy of investigation. In this study, left anterior descending (LAD) coronary artery was ligated to induce the CAD models in vivo, and cardiac functions were examined using echocardiography. Human umbilical vein endothelial cells (HUVEC) were subjected to H2O2-induced oxidative stress in vitro. The effects of DQP on CAD rat models and in vitro HUVEC were detected. Our results showed that DQP had cardio-protective effects in rat model. The intensity of capillaries in the marginal area of infarction of the rat heart was increased remarkably in DQP group, and the expression of PPARα and VEGF-2 were increased. The key enzymes involved in the transportation and intake of fatty acids, including CPT1A and CD36, both increased. In H2O2-induced endothelial cells injury models, DQP also showed protective roles and promoted capillary-like tube formation. DQP up-regulated key enzymes in fatty acids oxidation in H2O2-treated HUVEC. In addition, inhibition of CPT1A compromised the pro-angiogenic effects of DQP. In conclusion, fatty acids oxidation axis PPARα-CD36-CPT1A was involved in the pro-angiogenic roles of DQP against CAD. Cardiac CPT1A may serve as a target in therapeutic angiogenesis in clinics.
研究领域:
心脏病学
·
药理学
·
分子生物学
·
中医
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）