Purpurin 18 derivatives with a polyethylene glycol (PEG) linker were synthesized as novel photosensitizers (PSs) with the goal of using them in photodynamic therapy (PDT) for cancer. These compounds, derived from a second-generation PS, exhibit absorption at long wavelengths; considerable singlet oxygen generation and, in contrast to purpurin 18, have higher hydrophilicity due to decreased logP. Together, these properties make them potentially ideal PSs. To verify this, we screened the developed compounds for cell uptake, intracellular localization, antitumor activity and induced cell death type. All of the tested compounds were taken up into cancer cells of various origin and localized in organelles known to be important PDT targets, specifically, mitochondria and the endoplasmic reticulum. The incorporation of a zinc ion and PEGylation significantly enhanced the photosensitizing efficacy, decreasing IC50 (half maximal inhibitory compound concentration) in HeLa cells by up to 170 times compared with the parental purpurin 18. At effective PDT concentrations, the predominant type of induced cell death was apoptosis. Overall, our results show that the PEGylated derivatives presented have significant potential as novel PSs with substantially augmented phototoxicity for application in the PDT of cervical, prostate, pancreatic and breast cancer.
- MeSH
- fluorescenční mikroskopie MeSH
- fotochemoterapie metody MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- porfyriny chemie MeSH
- průtoková cytometrie MeSH
- rozpustnost MeSH
- singletový kyslík chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Photodynamic therapy has become a feasible direction for the treatment of both malignant and non-malignant diseases. It has been in the spotlight since FDA regulatory approval was granted to several photosensitizers worldwide. Nevertheless, there are still strong limitations in the targeting specificity that is vital to prevent systemic toxicity. Here, we report the synthesis and biological evaluation of a novel bimodal oxime conjugate composed of a photosensitizing drug, red-emitting pheophorbide a, and nandrolone (NT), a steroid specifically binding the androgen receptor (AR) commonly overexpressed in various tumors. We characterized the physico-chemical properties of the NT-pheophorbide a conjugate (NT-Pba) and singlet oxygen generation. Because light-triggered therapies have the potential to provide important advances in the treatment of hormone-sensitive cancer, the biological potential of this novel specifically-targeted photosensitizer was assessed in prostatic cancer cell lines in vitro using an AR-positive (LNCaP) and an AR-negative/positive cell line (PC-3). U-2 OS cells, both with and without stable AR expression, were used as a second cell line model. Interestingly, we found that the NT-Pba conjugate was not only photodynamically active and AR-specific, but also that its phototoxic effect was more pronounced compared to pristine pheophorbide a. We also examined the intracellular localization of NT-Pba. Live-cell fluorescence microscopy provided clear evidence that the NT-Pba conjugate localized in the endoplasmic reticulum and mitochondria. Moreover, we performed a competitive localization study with the excess of nonfluorescent NT, which was able to displace fluorescent NT-Pba from the cell interior, thereby further confirming the binding specificity. The oxime ether bond degradation was assayed in living cells by both real-time microscopy and a steroid receptor reporter assay using AR U-2 OS cells. Thus, NT-Pba is a promising candidate for both the selective targeting and eradication of AR-positive malignant cells by photodynamic therapy.
- MeSH
- chlorofyl analogy a deriváty chemie farmakologie MeSH
- fluorescenční mikroskopie MeSH
- fotochemoterapie * MeSH
- fotosenzibilizující látky chemická syntéza chemie farmakologie MeSH
- lidé MeSH
- molekulární struktura MeSH
- nádorové buněčné linie MeSH
- optické zobrazování MeSH
- oximy chemie farmakologie MeSH
- povrchové vlastnosti MeSH
- proliferace buněk účinky léků MeSH
- protinádorové látky chemická syntéza chemie farmakologie MeSH
- screeningové testy protinádorových léčiv MeSH
- testosteron analogy a deriváty chemie farmakologie MeSH
- velikost částic MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Drug targeting using functionalized nanoparticles to advance their transport to the dedicated site became a new standard in novel anticancer methods Anticancer photodynamic therapy also takes benefit from using nanoparticles by means of increasing targeting efficiency and decreased side effect. With this in mind, the silica-based nanoparticles, as drug delivery systems for the second-generation photosensitizer 5,10,15,20-tetrakis(m-hydroxyphenyl) chlorin (temoporfin) were developed. METHODS: In order to determine the stability and therapeutic performance of the selected nanomaterials in physiological fluids, their physicochemical properties (i.e. size, polydispersity, zeta potential) were measured by dynamic light scattering technique and the diameter and the morphology of the individual particles were visualized by a transmission electron microscopy. Their efficacy was compared with commercial temoporfin formulation in terms of in vitro phototoxicity in 4T1 (murine mammary carcinoma) and of in vivo anticancer effect in Nu/Nu mice bearing MDA-MB-231 tumors. RESULTS AND CONCLUSIONS: The two types of silica nanoparticles, porous and non-porous and with different surface chemical modification, were involved and critically compared within the study. Their efficacy was successfully demonstrated and was shown to be superior in comparison with commercial temoporfin formulation in terms of in vitro phototoxicity and cellular uptake as well as in terms of in vivo anticancer effect on human breast cancer model. Temoporfin-loaded silica nanoparticles also passed through the blood-brain barrier showing potential for the treatment of brain metastases.
- MeSH
- fotochemoterapie metody MeSH
- fotosenzibilizující látky aplikace a dávkování farmakologie MeSH
- lidé MeSH
- mesoporfyriny aplikace a dávkování farmakologie MeSH
- myši nahé MeSH
- nádorové buněčné linie MeSH
- nanočástice chemie MeSH
- nosiče léků chemie MeSH
- oxid křemičitý chemie MeSH
- polyethylenglykoly chemie MeSH
- transmisní elektronová mikroskopie MeSH
- uvolňování léčiv MeSH
- velikost částic MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The blood-brain barrier prevents the passage of many drugs that target the central nervous system. This paper presents the preparation and characterization of silica-based nanocarriers loaded with piracetam, pentoxifylline, and pyridoxine (drugs from the class of nootropics), which are designed to enhance the permeation of the drugs from the circulatory system through the blood-brain barrier. Their permeation was compared with non-nanoparticle drug substances (bulk materials) by means of an in vivo model of rat brain perfusion. The size and morphology of the nanoparticles were characterized by transmission electron microscopy. The content of the drug substances in silica-based nanocarriers was analysed by elemental analysis and UV spectrometry. Microscopic analysis of visualized silica nanocarriers in the perfused brain tissue was performed. The concentration of the drug substances in the tissue was determined by means of UHPLC-DAD/HRMS LTQ Orbitrap XL. It was found that the drug substances in silica-based nanocarriers permeated through the blood brain barrier to the brain tissue, whereas bulk materials were not detected in the brain.
- MeSH
- hematoencefalická bariéra metabolismus MeSH
- krysa rodu rattus MeSH
- nanočástice chemie MeSH
- nootropní látky * chemie farmakokinetika farmakologie MeSH
- nosiče léků * chemie farmakokinetika farmakologie MeSH
- oxid křemičitý * chemie farmakokinetika farmakologie MeSH
- potkani Wistar MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The rational design of molecules with selective intracellular targeting is a great challenge for contemporary chemistry and life sciences. Here, we demonstrate a rational approach to development of compartment-specific fluorescent dyes from the γ-aryl substituted pentamethine family. These novel dyes exhibit an extraordinary affinity and selectivity for cardiolipin in inner mitochondrial membrane and possess excellent photostability, fluorescent properties, and low phototoxicity. Selective imaging of live and fixed mitochondria was achieved in various cell lines using nanomolar concentrations of these dyes. Their high localization specificity and low toxicity enables study of morphological changes, structural complexity, and dynamics of mitochondria playing a pivotal role in many pathological diseases. These far-red emitting dyes could also serve in a variety of biomedical applications.
- MeSH
- buněčné linie MeSH
- DNA analýza MeSH
- fluorescenční barviva analýza metabolismus MeSH
- kardiolipiny analýza metabolismus MeSH
- krystalografie rentgenová MeSH
- kultivované buňky MeSH
- kur domácí MeSH
- lidé MeSH
- ligandy MeSH
- mitochondrie metabolismus ultrastruktura MeSH
- molekulární modely MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
FTIR, circular dichroism (CD) and fluorescence spectroscopies were used to characterize conformational changes in horse liver alcohol dehydrogenase (HLADH) and ketoreductase (KRED 117) upon physical and covalent immobilizations on silica particles (functionalized with amino, epoxy and thiol groups) of different sizes. Conformational changes for immobilized enzymes were associated with high and low frequency shifts of the amide I and II bands. CD spectra of native HLADH and KRED 117 characterized with a negative peak at 222nm indicating a α-helical structure. The disappearance of the negative peak in the CD spectra of immobilized enzymes and appearance of a positive peak at 222nm supported these observations. These findings demonstrated unfolding of folded enzymes and exposure of the amino acid residues during denaturation with a red shift in tryptophan fluorescence. The decrease in specific activities (by 60-70% in all cases) for both immobilized enzymes was correlated to those of conformational changes. Silica-attached enzyme-NADH systems were evaluated for enantioselective reduction of 1-(p-methoxyphenyl)-propan-2-one. Conformational changes enhanced the enantioselectivity of immobilized HLADH with a switch in its stereoselectivity. In the case of immobilized KRED 117, kinetic values (V(max) and K(m)) were lower than that of the free enzyme, without enhancing enzyme enantio- and stereoselectivity.
- MeSH
- alkoholdehydrogenasa chemie MeSH
- alkoholoxidoreduktasy chemie MeSH
- cirkulární dichroismus MeSH
- enzymy imobilizované chemie MeSH
- fluorescenční spektrometrie MeSH
- játra enzymologie MeSH
- kinetika MeSH
- koenzymy chemie MeSH
- koně MeSH
- NAD chemie MeSH
- nanočástice chemie ultrastruktura MeSH
- oxid křemičitý chemie MeSH
- rozbalení proteinů MeSH
- sekundární struktura proteinů MeSH
- spektroskopie infračervená s Fourierovou transformací MeSH
- stereoizomerie MeSH
- substrátová specifita MeSH
- terciární struktura proteinů MeSH
- tryptofan chemie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The review article is devoted mainly to the description of applications of gold nanoparticles (GNPs) in separation sciences, especially in electromigration and chromatographic techniques. The applications of GNPs in particular separation methods, CE, microchip CE, MEKC, CEC, HPLC and GC, are classified according to the molecular size of the analytes from low-molecular-mass compounds via medium sized substances to biopolymers (proteins and nucleic acids). A very recent and promising utilization of GNPs for sample preparation, preconcentration and preseparation of selected analytes from complex matrices is presented as well. Moreover, in two introductory sections, typical preparation procedures of the GNPs and their modifications are presented and physico-chemical and analytical methods employed for characterization of the native and modified GNPs are briefly introduced.
- MeSH
- chemická frakcionace přístrojové vybavení metody MeSH
- chemické techniky analytické přístrojové vybavení metody MeSH
- chromatografie micelární elektrokinetická kapilární MeSH
- chromatografie plynová MeSH
- elektroforéza kapilární MeSH
- kovové nanočástice chemie MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- zlato MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Two porphyrin-brucine quaternary ammonium salts were immobilized on gold nanoparticles and their suitability for both in vitro and in vivo photodynamic therapy (PDT) was assayed using the basaloid squamous cell carcinoma PE/CA-PJ34 cell line. In vitro PDT experiments revealed that the gold nanoparticle-bound conjugates were less effective than unbound conjugates in killing cells. However, the same conjugates were more effective in reducing tumor size in vivo, with complete tumor regression observed.
- MeSH
- alkylace MeSH
- biologický transport MeSH
- buněčná smrt účinky léků účinky záření MeSH
- fotochemoterapie metody MeSH
- intracelulární prostor metabolismus MeSH
- kovové nanočástice chemie MeSH
- lidé MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádory farmakoterapie MeSH
- porfyriny chemie MeSH
- rozpouštědla chemie MeSH
- strychnin analogy a deriváty chemie metabolismus farmakologie terapeutické užití MeSH
- zlato chemie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
