protein nanocarriers Dotaz Zobrazit nápovědu
Anthracycline antibiotic drugs are widely used in treatment of many patients with cancer. First anthracycline drug, daunomycin (or daunorubicin) was found in a number of different wild type strains of Streptomyces. However, in cancer treatment the most commonly used anthracycline drug is doxorubicin (DOX) or one of its 2000 known analogs1. DOX is used in treatment of many different types of cancer such as neuroblastomas, leukaemia, lymphomas or breast, testicle, ovarian, lung, bladder, thyroid gland or head and neck carcinomas. Although it is so widely used, many side effects have been observed in patients such as sores in mouth and on lips, darkening of palms and nails, unusual bleeding and bruising, nausea and vomiting, and life-threatening cardiotoxicity 2. To eliminate the negative side effects of cancer treatment, researchers are trying to find either new analogs of DOX which are non-toxic for healthy cells or new way to deliver DOX directly into the cancer cells. For targeted delivery, it is possible to administer the drug directly into solid tumor. However, non-solid tumors or tumors with unknown location in patient’s body require encapsulation of DOX in suitable nanocarrier. Liposomal form of DOX is already being sold under the trade name Myocet 3. For enhanced biocompatibility, the liposomes were modified with polyethylene glycol under the trade name Doxil 4. Protein based natural nanocarriers in comparison with artificial nanocarriers seem to be more suitable for delivery of the drugs in patient’s body because of their lower immune response. The protein nanocarriers are usually self-assembled and can either be protein cages, viral capsids or virus-like particles 5. In this work, we compared two types of protein nanocarriers – phage λ and apoferritin, by their ability to encapsulate anthracycline drug doxorubicine. The encapsulation was verified by fluorescence of doxorubicin after the removal of free, non-encapsulated doxorubicin by dialysis.
- Klíčová slova
- enkapsulace léčiv,
- MeSH
- apoferritiny MeSH
- bakteriofág lambda MeSH
- doxorubicin * MeSH
- fluorescence MeSH
- nanomedicína MeSH
- nanostruktury MeSH
- systémy cílené aplikace léků * MeSH
- Publikační typ
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Several plasminogen activators (PAs) have been found effective in treating different thromboembolic diseases. However, administration of conventional thrombolytic therapy is limited by a low efficacy of present formulations of PAs. Conventional treatments using these therapeutic proteins are associated with several limitations including rapid inactivation and clearance, short half-life, bleeding complications or non-specific tissue targeting. Liposome-based formulations of PAs such as streptokinase, tissue-plasminogen activator and urokinase have been developed to improve the therapeutic efficacy of these proteins. Resulting liposomal formulations were found to preserve the original activity of PAs, promote their selective delivery and improve thrombus targeting. Therapeutic potential of these liposome-based PAs has been demonstrated successfully in various pre-clinical models in vivo. Reductions in unwanted side effects (e.g., hemorrhage or immunogenicity) as well as enhancements of efficacy and safety were achieved in comparison to currently existing treatment options based on conventional formulations of PAs. This review summarizes present achievements in: (i) preparation of liposome-based formulations of various PAs, (ii) development of PEGylated and targeted liposomal PAs, (iii) physico-chemical characterization of these developed systems, and (iv) testing of their thrombolytic efficacy. We also look to the future and the imminent arrival of theranostic liposomal formulations to move this field forward.
- MeSH
- aktivátor plasminogenu urokinasového typu aplikace a dávkování terapeutické užití MeSH
- fibrinolytika aplikace a dávkování terapeutické užití MeSH
- lidé MeSH
- liposomy chemie ultrastruktura MeSH
- metaloendopeptidasy aplikace a dávkování terapeutické užití MeSH
- nanostruktury chemie ultrastruktura MeSH
- plasmin aplikace a dávkování terapeutické užití MeSH
- streptokinasa aplikace a dávkování terapeutické užití MeSH
- tkáňový aktivátor plazminogenu aplikace a dávkování terapeutické užití MeSH
- tromboembolie farmakoterapie MeSH
- trombolytická terapie metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Glycogen-nucleic acid constructs i.e., glycoplexes are emerging promising platforms for the alteration of gene expression and transcription. Understanding the interaction of glycoplexes with human blood components, such as serum proteins and peripheral blood mononuclear cells (PBMCs), is important to overcome immune cell activation and control biodistribution upon administration of the glycoplexes in vivo. Herein, we investigated the interactions of polyethylene glycol (PEG)ylated and non-PEGylated glycoplexes carrying siRNA molecules with PBMCs isolated from the blood of healthy donors. We found that both types of glycoplexes were non-toxic and were primarily phagocytosed by monocytes without triggering a pro-inflammatory interleukin 6 cytokine production. Furthermore, we investigated the role of the protein corona on controlling the internalization efficiency in immune cells - we found that the adsorption of serum proteins, in particular haptoglobin, alpha-1-antitrypsin and apolipoprotein A-II, onto the non-PEGylated glycoplexes, significantly reduced the uptake of the glycoplexes by PBMCs. Moreover, the non-PEGylated glycoplexes were efficient in the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) knockdown in monocytic THP-1 cell line. This study provides an insight into the rational design of glycogen-based nanocarriers for the safe delivery of siRNA without eliciting unwanted immune cell activation and efficient siRNA activity upon its delivery.
Allyl- and propargyl ethers of umbelliferone are sensitive probes for palladium and platinum, including anticancer compounds cisplatin, carboplatin and oxaliplatin, and effective for direct visualization of protein and DNA complexes with organometallic compounds in polyacrylamide gels allowing easy detection of interactions with analyzed protein or nucleic acid. Both probes can be used for fast evaluation of Pd/Pt binding to nanocarriers relevant in drug targeted therapy or specific clinically relevant target macromolecules.
In this study, we discussed the use of bacteriophage λ as a nanocarrier of anthracycline drug doxorubicin. This drug is used in treatment of cancer, which is the cause of death of every fourth patient in developed countries. Doxorubicin has many severe side effects for patients' healthy cells, which lowers their well-being. These can be eliminated by encapsulation of doxorubicin into suitable nanocarrier, such as bacteriophage λ capsid. In this work, infusion method was used for this encapsulation and proven by absorbance and fluorescence measurement of whole bacteriophage. It was concluded, that bacteriophage both intercalates into DNA and binds to capsid proteins of bacteriophage λ.
- Klíčová slova
- enkapsulace léčiv,
- MeSH
- bakteriofág lambda * MeSH
- DNA MeSH
- doxorubicin * MeSH
- fluorescenční spektrometrie statistika a číselné údaje MeSH
- nanočástice MeSH
- systémy cílené aplikace léků * MeSH
- virové plášťové proteiny MeSH
- virům podobné částice * MeSH
- Publikační typ
- hodnotící studie MeSH
- práce podpořená grantem MeSH
Routine cancer treatment often causes damage to whole human organism, because cytostatic drugs, which are used during the treatment and whose function is to stop growth of cancer cells, affect also healthy cells and untargeted tissues. Due to this fact, nanocarriers and targeted drug delivery have been recently studied. Side effects and organ damage can be reduced by using nanocarriers and targeted drug delivery. The aim of this assay was to monitor and characterize apoferritin as a nanocarrier for targeted drug delivery. In this work the surface of nanocarrier composed of apoferritin with encapsulated doxorubicin was modified with gold nanoparticles or chloroauric acid, conjugated with linker (HWR peptide) because of correct binding of the antibody and labeled with specific antibody. In this assay the resulting nanocarrier was studied with gel electrophoresis and fluorescent methods.
- Klíčová slova
- nanotransportér, teranostika,
- MeSH
- apoferritiny * MeSH
- doxorubicin MeSH
- elektroforéza MeSH
- ELISA MeSH
- fluorescenční protilátková technika MeSH
- imunoglobulin G MeSH
- kovové nanočástice * MeSH
- nanomedicína MeSH
- protilátky MeSH
- systémy cílené aplikace léků * MeSH
- zlato MeSH
- Publikační typ
- práce podpořená grantem MeSH
RNA interference by short interfering RNAs (siRNAs) holds promise as a therapeutic strategy, but use of siRNAs in vivo remains limited. Here, we developed a system to target delivery of siRNAs to glomeruli via poly(ethylene glycol)-poly(l-lysine)-based vehicles. The siRNA/nanocarrier complex was approximately 10 to 20 nm in diameter, a size that would allow it to move across the fenestrated endothelium to access to the mesangium. After intraperitoneal injection of fluorescence-labeled siRNA/nanocarrier complexes, we detected siRNAs in the blood circulation for a prolonged time. Repeated intraperitoneal administration of a mitogen-activated protein kinase 1 (MAPK1) siRNA/nanocarrier complex suppressed glomerular MAPK1 mRNA and protein expression in a mouse model of glomerulonephritis; this improved kidney function, reduced proteinuria, and ameliorated glomerular sclerosis. Furthermore, this therapy reduced the expression of the profibrotic markers TGF-beta1, plasminogen activator inhibitor-1, and fibronectin. In conclusion, we successfully silenced intraglomerular genes with siRNA using nanocarriers. This technique could aid the investigation of molecular mechanisms of renal disease and has potential as a molecular therapy of glomerular diseases.
Due to many adverse effects of conventional chemotherapy, novel methods of targeting drugs to cancer cells are being investigated. Nanosize carriers are a suitable platform for this specific delivery. Herein, we evaluated the long-term stability of the naturally found protein nanocarrier apoferritin (Apo) with encapsulated doxorubicin (Dox). The encapsulation was performed using Apo's ability to disassemble reversibly into its subunits at low pH (2.7) and reassemble in neutral pH (7.2), physically entrapping drug molecules in its cavity (creating ApoDox). In this study, ApoDox was prepared in water and phosphate-buffered saline and stored for 12 weeks in various conditions (-20°C, 4°C, 20°C, and 37°C in dark, and 4°C and 20°C under ambient light). During storage, a very low amount of prematurely released drug molecules were detected (maximum of 7.5% for ApoDox prepared in PBS and 4.4% for ApoDox prepared in water). Fourier-transform infrared spectra revealed no significant differences in any of the samples after storage. Most of the ApoDox prepared in phosphate-buffered saline and ApoDox prepared in water and stored at -20°C formed very large aggregates (up to 487% of original size). Only ApoDox prepared in water and stored at 4°C showed no significant increase in size or shape. Although this storage caused slower internalization to LNCaP prostate cancer cells, ApoDox (2.5 μM of Dox) still retained its ability to inhibit completely the growth of 1.5×10(4) LNCaP cells after 72 hours. ApoDox stored at 20°C and 37°C in water was not able to deliver Dox inside the nucleus, and thus did not inhibit the growth of the LNCaP cells. Overall, our study demonstrates that ApoDox has very good stability over the course of 12 weeks when stored properly (at 4°C), and is thus suitable for use as a nanocarrier in the specific delivery of anticancer drugs to patients.
- MeSH
- antitumorózní látky aplikace a dávkování farmakokinetika farmakologie MeSH
- apoferritiny aplikace a dávkování chemie farmakokinetika MeSH
- doxorubicin aplikace a dávkování chemie MeSH
- koncentrace vodíkových iontů MeSH
- léky antitumorózní - screeningové testy MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory prostaty farmakoterapie patologie MeSH
- nosiče léků aplikace a dávkování chemie farmakokinetika MeSH
- stabilita léku MeSH
- systémy cílené aplikace léků MeSH
- uvolňování léčiv MeSH
- voda chemie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Gadolinium (Gd)-based contrast agents are extensively used for magnetic resonance imaging (MRI). Liposomes are potential nanocarrier-based biocompatible platforms for development of new generations of MRI diagnostics. Liposomes with Gd-complexes (Gd-lip) co-encapsulated with thrombolytic agents can serve both for imaging and treatment of various pathological states including stroke. In this study, we evaluated nanosafety of Gd-lip containing PE-DTPA chelating Gd+3 prepared by lipid film hydration method. We detected no cytotoxicity of Gd-lip in human liver cells including cancer HepG2, progenitor (non-differentiated) HepaRG, and differentiated HepaRG cells. Furthermore, no potential side effects of Gd-lip were found using a complex system including general biomarkers of toxicity, such as induction of early response genes, oxidative, heat shock and endoplasmic reticulum stress, DNA damage responses, induction of xenobiotic metabolizing enzymes, and changes in sphingolipid metabolism in differentiated HepaRG. Moreover, Gd-lip did not show pro-inflammatory effects, as assessed in an assay based on activation of inflammasome NLRP3 in a model of human macrophages, and release of eicosanoids from HepaRG cells. In conclusion, this in vitro study indicates potential in vivo safety of Gd-lip with respect to hepatotoxicity and immunopathology caused by inflammation.
- MeSH
- diethylentriaminpentaacetát gadolinia * škodlivé účinky toxicita MeSH
- fibrinolytika MeSH
- fosfatidylethanolaminy * škodlivé účinky toxicita MeSH
- hepatocyty účinky léků MeSH
- inflamasomy MeSH
- kontrastní látky * MeSH
- kultivované buňky MeSH
- lidé MeSH
- liposomy * MeSH
- magnetická rezonanční tomografie * MeSH
- makrofágy účinky léků MeSH
- nanočástice MeSH
- nosiče léků * MeSH
- protein NLRP3 MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
To improve the treatment of psoriasiform inflammation, we developed actively targeted nanocarriers loaded with the phosphodiesterase 4 inhibitor AN2728. Methods: Phospholipid-poly(lactic-co-glycolic acid) nanohybrids were prepared and conjugated with monovalent anti-desmoglein 3 antibody to bind keratinocytes. Results: The actively targeted nanohybrids were 229 nm in mean size with a nearly neutral surface charge. Flow cytometry and confocal microscopy showed a 9-fold increase in keratinocyte uptake of targeted nanohybrids relative to non-targeted nanoparticles. The nanoparticles localized mainly in lysosomes after internalization. AN2728-loaded antibody-conjugated nanocarriers inhibited cytokine/chemokine overexpression in activated keratinocytes without affecting cell viability. The targeted nanohybrids also suppressed neutrophil migration by reducing CXCL1 and CXCL2 release from keratinocytes. Following subcutaneous administration in mice, the nanohybrids distributed to the epidermis and hair follicles. In a psoriasis-like skin mouse model, the actively targeted nanoparticles were superior to free drug and non-targeted nanoparticles in mitigating skin inflammation. Intervention with the targeted nanosystem reduced the epidermal thickness of the psoriasiform lesion from 191 to 42 µm, decreased the Psoriasis Area Severity Index by 74%, restored barrier function, and returned chemokine levels to baseline. Conclusions: Our developed nanosystem was safe and demonstrated efficient targeting properties for the treatment of cutaneous inflammation.
- MeSH
- bicyklické sloučeniny heterocyklické aplikace a dávkování farmakologie MeSH
- buněčné linie keratinocytů HaCaT MeSH
- chemokin CXCL1 účinky léků imunologie MeSH
- chemokin CXCL2 účinky léků imunologie MeSH
- chemotaxe účinky léků MeSH
- desmoglein 3 imunologie MeSH
- epidermis MeSH
- fosfolipidy * MeSH
- imunokonjugáty farmakologie MeSH
- inhibitory fosfodiesterasy 4 aplikace a dávkování farmakologie MeSH
- keratinocyty účinky léků imunologie MeSH
- kopolymer kyseliny glykolové a mléčné * MeSH
- lidé MeSH
- lyzozomy metabolismus ultrastruktura MeSH
- modely nemocí na zvířatech MeSH
- myši MeSH
- nanočástice * MeSH
- neutrofily účinky léků MeSH
- nosiče léků MeSH
- protilátky imunologie MeSH
- psoriáza imunologie patologie MeSH
- sloučeniny boru aplikace a dávkování farmakologie MeSH
- systémy cílené aplikace léků MeSH
- vlasový folikul MeSH
- zánět MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
