The tear fluids from three healthy individuals and three patients with diabetes mellitus were examined using atomic force microscopy-infrared spectroscopy (AFM-IR) and Fourier transform infrared spectroscopy (FTIR). The dried tear samples showed different surface morphologies: the control sample had a dense network of heart-shaped dendrites, while the diabetic sample had fern-shaped dendrites. By using the AFM-IR technique we identified spatial distribution of constituents, indicating how diabetes affects the structural characteristics of dried tears. FTIR showed that the dendritic structures gradually disappeared over time due to glucose-induced lysozyme damage. The tear fluid from diabetes mellitus patients has a higher concentration of glucose, which accelerates the breakdown of lysozyme and, as a result, the quick loss of the dendritic structure. Our study shows that analysis of dry tear fluid can be promising technique for the detection of glycated proteins that reveal long lasting hyperglycemia and diabetes mellitus.
- Klíčová slova
- Diabetes mellitus, Photothermal AFM-IR analysis, Tear fluid,
- MeSH
- biologické markery analýza metabolismus MeSH
- diabetes mellitus * metabolismus diagnóza MeSH
- dospělí MeSH
- glukosa metabolismus MeSH
- lidé středního věku MeSH
- lidé MeSH
- mikroskopie atomárních sil * metody MeSH
- muramidasa metabolismus MeSH
- slzy * chemie metabolismus MeSH
- spektroskopie infračervená s Fourierovou transformací metody MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- biologické markery MeSH
- glukosa MeSH
- muramidasa MeSH
We recently prepared pH-responsive HPMA copolymer conjugates of bradykinin (P-BK), which release BK in response to the acidic tumor microenvironment, and found that administration of P-BK increased the tumor accumulation and therapeutic efficacy of nanomedicine. Because the release of BK from P-BK determines its onset of action, P-BKs with different release rates were prepared, and their properties were evaluated. The release kinetics were significantly altered by substitution proximal to hydrazone bond, release constant of methyl-substituted P-BK (P-MeBK) was approximately 4- and 80-fold higher than that of cyclopropyl-substituted P-BK (P-CPBK) and phenyl-substituted P-BK (P-PhBK). None of the P-BKs were active, but the release of BK restored their BK-like activity. Pre-administration of the P-BKs increased the tumor accumulation of nanomedicine in C26 tumor-bearing mice by 2- and 1.4-fold for P-MeBK and P-PhBK at 3 and 6 h. Altogether, this study provides insights into the design of pH-responsive nanodrugs with the desired release properties to target acidic lesions such as cancer and inflammation.
- Klíčová slova
- Bradykinin, Controlled release, HPMA polymer, Hydrazone, Tumor,
- MeSH
- bradykinin MeSH
- doxorubicin chemie MeSH
- koncentrace vodíkových iontů MeSH
- myši MeSH
- nádorové mikroprostředí MeSH
- nádory * farmakoterapie MeSH
- nanomedicína MeSH
- polymery * chemie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- bradykinin MeSH
- doxorubicin MeSH
- polymery * MeSH
We synthesized three novel STAT3 inhibitors (S3iD1-S3iD3) possessing oxoheptanoic residue enabling linkage to HPMA copolymer carrier via a pH-sensitive hydrazone bond. HPMA copolymer conjugates bearing doxorubicin (Dox) and our STAT3 inhibitors were synthesized to evaluate the anticancer effect of Dox and STAT3 inhibitor co-delivery into tumors. S3iD1-3 and their copolymer-bound counterparts (P-S3iD1-P-S3iD3) showed considerable in vitro cytostatic activities in five mouse and human cancer cell lines with IC50 ~0.6-7.9 μM and 0.7-10.9 μM, respectively. S3iD2 and S3iD3 were confirmed to inhibit the STAT3 signaling pathway. The combination of HPMA copolymer-bound Dox (P-Dox) and P-S3iD3 at the dosage showing negligible toxicity demonstrated significant antitumor activity in B16F10 melanoma-bearing mice and completely cured 2 out of 15 mice. P-Dox alone had a significantly lower therapeutic activity with no completely cured mice. Thus, polymer conjugates bearing STAT3 inhibitors may be used for the chemosensitization of chemorefractory tumors.
- Klíčová slova
- Doxorubicin, HPMA copolymer carrier, STAT3 inhibitor, pH-sensitive drug release,
- MeSH
- doxorubicin * farmakologie terapeutické užití MeSH
- koncentrace vodíkových iontů MeSH
- kyseliny polymethakrylové MeSH
- lidé MeSH
- methakryláty * MeSH
- myši MeSH
- nádory * farmakoterapie MeSH
- transkripční faktor STAT3 metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- doxorubicin-N-(2-hydroxypropyl)methacrylamide copolymer conjugate MeSH Prohlížeč
- doxorubicin * MeSH
- hydroxypropyl methacrylate MeSH Prohlížeč
- kyseliny polymethakrylové MeSH
- methakryláty * MeSH
- STAT3 protein, human MeSH Prohlížeč
- transkripční faktor STAT3 MeSH
Rheumatoid arthritis is a chronic inflammatory autoimmune disease caused by alteration of the immune system. Current therapies have several limitations and the use of nanomedicines represents a promising strategy to overcome them. By employing a mouse model of adjuvant induced arthritis, we aimed to evaluate the biodistribution and therapeutic effects of glucocorticoid dexamethasone conjugated to a nanocarrier based on biocompatible N-(2-hydroxypropyl) methacrylamide copolymers. We observed an increased accumulation of dexamethasone polymer nanomedicines in the arthritic mouse paw using non-invasive fluorescent in vivo imaging and confirmed it by the analysis of tissue homogenates. The dexamethasone conjugate exhibited a dose-dependent healing effect on arthritis and an improved therapeutic outcome compared to free dexamethasone. Particularly, significant reduction of accumulation of RA mediator RANKL was observed. Overall, our data suggest that the conjugation of dexamethasone to a polymer nanocarrier by means of stimuli-sensitive spacer is suitable strategy for improving rheumatoid arthritis therapy.
- Klíčová slova
- Biodistribution, Dexamethasone, HPMA, RANKL, Rheumatoid arthritis,
- MeSH
- artritida experimentální farmakoterapie patologie MeSH
- dexamethason * chemie farmakokinetika aplikace a dávkování farmakologie terapeutické užití MeSH
- myši MeSH
- nanočástice chemie MeSH
- nosiče léků chemie farmakokinetika MeSH
- polymery * chemie farmakokinetika MeSH
- revmatoidní artritida * farmakoterapie patologie MeSH
- tkáňová distribuce MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- dexamethason * MeSH
- nosiče léků MeSH
- polymery * MeSH
Biodistribution analyses of nanocarriers are often performed with optical imaging. Though dye tags can interact with transporters, e.g., organic anion transporting polypeptides (OATPs), their influence on biodistribution was hardly studied. Therefore, this study compared tumor cell uptake and biodistribution (in A431 tumor-bearing mice) of four near-infrared fluorescent dyes (AF750, IRDye750, Cy7, DY-750) and dye-labeled poly(N-(2-hydroxypropyl)methacrylamide)-based nanocarriers (dye-pHPMAs). Tumor cell uptake of hydrophobic dyes (Cy7, DY-750) was higher than that of hydrophilic dyes (AF750, IRDye750), and was actively mediated but not related to OATPs. Free dyes' elimination depended on their hydrophobicity, and tumor uptake correlated with blood circulation times. Dye-pHPMAs circulated longer and accumulated stronger in tumors than free dyes. Dye labeling significantly influenced nanocarriers' tumor accumulation and biodistribution. Therefore, low-interference dyes and further exploration of dye tags are required to achieve the most unbiased results possible. In our assessment, AF750 and IRDye750 best qualified for labeling hydrophilic nanocarriers.
- Klíčová slova
- Biodistribution, Drug delivery system, Fluorescent dye-labeling, Molecular imaging, Optical imaging,
- MeSH
- fluorescenční barviva chemie MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádory * diagnostické zobrazování farmakoterapie MeSH
- nosiče léků * chemie MeSH
- optické zobrazování MeSH
- tkáňová distribuce MeSH
- zkreslení výsledků (epidemiologie) MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- fluorescenční barviva MeSH
- nosiče léků * MeSH
In this study, we developed a nanoformulation of 5-aminolevulinic acid (5-ALA) for tumor-targeted photodynamic therapy, in which 5-ALA was conjugated with a biocompatible polymer N-(2-hydroxypropyl)methacrylamide (HPMA) through the hydrazone bond, i.e., P-ALA. P-ALA behaves as the nano-sized molecule with an average size of 5.5 nm in aqueous solution. P-ALA shows a largely increased release rate in acidic pH than physiological pH, suggesting the rapid release profile in acidic tumor environment. P-ALA did not show apparent cytotoxicity up to 0.1 mg/ml, however, under light irradiation, remarkable cell death was induced with the IC50 of 20-30 μg/ml. More importantly, we found significantly higher tumor accumulation of P-ALA than 5-ALA which benefit from its nano-size by taking advantage of the enhanced permeability and retention (EPR) effect. Consequently, P-ALA exhibited much improved in vivo antitumor efficacy without any apparent side effects. We thus anticipate the application of P-ALA as a nano-designed photosensitizer for anticancer photodynamic therapy.
- Klíčová slova
- 5-Aminolevulinic acid, EPR effect, HPMA copolymer, Photodynamic therapy, Tumor targeting,
- MeSH
- doxorubicin farmakologie MeSH
- fotochemoterapie * MeSH
- kyselina aminolevulová farmakologie terapeutické užití MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory * patologie MeSH
- polymery chemie MeSH
- protinádorové látky * farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- doxorubicin MeSH
- hydroxypropyl methacrylate MeSH Prohlížeč
- kyselina aminolevulová MeSH
- polymery MeSH
- protinádorové látky * MeSH
Polymer nanomedicines with anti-tumor activity should exhibit sufficient stability during systemic circulation to the target tissue; however, they should release the active drug selectively in the tumor. Thus, choice of a tumor-specific stimuli-sensitive spacer between the drug and the carrier is critical. Here, a series of polymer conjugates of anti-cancer drugs doxorubicin and pirarubicin covalently bound to copolymers based on N-(2-hydroxypropyl)methacrylamide via various enzymatically cleavable oligopeptide spacers were prepared and characterized. The highest rate of the drug release from the polymer carriers in presence of the lysosomal protease cathepsin B was determined for the copolymers with Val-Cit-Aba spacer. Copolymers containing pirarubicin were more cytotoxic and showed higher internalization rate than the corresponding doxorubicin counterparts. The conjugates containing GFLG and Val-Cit-Aba spacers exhibited the highest anti-tumor efficacy in vivo against murine sarcoma S-180, the highest rate of the enzymatically catalyzed drug release, and the highest cytotoxicity in vitro.
- Klíčová slova
- Drug delivery, Enzymatic release, Polymer cancerostatics,
- MeSH
- doxorubicin farmakologie chemie MeSH
- lidé MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádory * farmakoterapie MeSH
- nanomedicína MeSH
- nosiče léků chemie MeSH
- polymery chemie MeSH
- protinádorové látky * farmakologie terapeutické užití chemie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- doxorubicin MeSH
- nosiče léků MeSH
- pirarubicin MeSH Prohlížeč
- polymery MeSH
- protinádorové látky * MeSH
- Klíčová slova
- Actinonin, Matrix metalloproteinases, Metastases, Polymer conjugate, pH-sensitive release,
- MeSH
- doxorubicin farmakologie MeSH
- kyseliny hydroxamové MeSH
- lidé MeSH
- methakryláty MeSH
- nádory * farmakoterapie MeSH
- nanomedicína * MeSH
- polymery farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- actinonin MeSH Prohlížeč
- doxorubicin MeSH
- hydroxypropyl methacrylate MeSH Prohlížeč
- kyseliny hydroxamové MeSH
- methakryláty MeSH
- polymery MeSH
Nanoparticles offer targeted delivery of drugs with minimal toxicity to surrounding healthy tissue and have great potential in the management of human papillomavirus (HPV)-related diseases. We synthesized lipid-modified AS1411 aptamers capable of forming nanoaggregates in solution containing Mg2+. The nanoaggregates presented suitable properties for pharmaceutical applications such as small size (100 nm), negative charge, and drug release. The nanoaggregates were loaded with acridine orange derivative C8 for its specific delivery into cervical cancer cell lines and HPV-positive tissue biopsies. This improved inhibition of HeLa proliferation and cell uptake without significantly affecting healthy cells. Finally, the nanoaggregates were incorporated in a gel formulation with promising tissue retention properties aiming at developing a local delivery strategy of the nanoaggregates in the female genital tract. Collectively, these findings suggest that the nanoformulation protocol has great potential for the delivery of both anticancer and antiviral agents, becoming a novel modality for cervical cancer management.
- Klíčová slova
- AS1411 G-quadruplex, Cervical cancer, Drug delivery, HPV, Nanoaggregates,
- MeSH
- antivirové látky * chemie farmakokinetika farmakologie MeSH
- aptamery nukleotidové * chemie farmakokinetika farmakologie MeSH
- HeLa buňky MeSH
- lékové transportní systémy * MeSH
- lidé MeSH
- nádory děložního čípku farmakoterapie metabolismus MeSH
- oligodeoxyribonukleotidy * chemie farmakokinetika farmakologie MeSH
- proliferace buněk účinky léků MeSH
- protinádorové látky * chemie farmakokinetika farmakologie MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- AGRO 100 MeSH Prohlížeč
- antivirové látky * MeSH
- aptamery nukleotidové * MeSH
- oligodeoxyribonukleotidy * MeSH
- protinádorové látky * MeSH
Liposomes functionalized with monoclonal antibodies or their antigen-binding fragments have attracted much attention as specific drug delivery devices for treatment of various diseases including cancer. The conjugation of antibodies to liposomes is usually achieved by covalent coupling using cross-linkers in a reaction that might adversely affect the characteristics of the final product. Here we present an alternative strategy for liposome functionalization: we created a recombinant Fab antibody fragment genetically fused on its C-terminus to the hydrophobic peptide derived from pulmonary surfactant protein D, which became inserted into the liposomal bilayer during liposomal preparation and anchored the Fab onto the liposome surface. The Fab-conjugated liposomes specifically recognized antigen-positive cells and efficiently delivered their cargo, the Alexa Fluor 647 dye, into target cells in vitro and in vivo. In conclusion, our approach offers the potential for straightforward development of nanomedicines functionalized with an antibody of choice without the need of harmful cross-linkers.
- Klíčová slova
- Active targeting, Antibody engineering, Immunoliposome, Liposome functionalization, Recombinant Fab antibody fragment,
- MeSH
- antigen CD48 metabolismus MeSH
- antigeny CD59 metabolismus MeSH
- imunoglobuliny - Fab fragmenty chemie imunologie metabolismus MeSH
- Jurkat buňky MeSH
- lidé MeSH
- liposomy chemie MeSH
- lymfom imunologie metabolismus patologie MeSH
- monoklonální protilátky chemie imunologie metabolismus MeSH
- myši MeSH
- nádorové buňky kultivované MeSH
- peptidové fragmenty imunologie metabolismus MeSH
- protein D asociovaný s plicním surfaktantem imunologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antigen CD48 MeSH
- antigeny CD59 MeSH
- CD48 protein, human MeSH Prohlížeč
- CD59 protein, human MeSH Prohlížeč
- imunoglobuliny - Fab fragmenty MeSH
- liposomy MeSH
- monoklonální protilátky MeSH
- peptidové fragmenty MeSH
- protein D asociovaný s plicním surfaktantem MeSH
