Novel BODIPY-estradiol conjugates have been synthesized by selecting position C-3-O for labeling. The conjugation strategy was based on Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) or etherification. Estradiol derivatives used as azide partners bearing an ω-azidoalkyl function through C4-C8-long linkers have been prepared. CuAAC reactions of estradiol azides with BODIPY alkyne furnished fluorescent 3-O-labeled conjugates bearing the triazole ring as a coupling moiety. Williamson etherifications of 3-O-(ω-bromoalkyl)-17β-estradiol derivatives with BODIPY-OH resulted in labeled conjugates connected with an ether moiety. Interactions of the conjugates with estrogen receptor (ER) were investigated using molecular docking calculations in comparison with estradiol. The conjugates occupied both the classical and alternative binding sites on human ERα, with slightly lower binding affinity to references estradiol and diethystilbestrol. All compounds have displayed reasonable estrogenic activity. They increased the proliferation of ER-positive breast cancer cell line MCF7 contrary to ER-negative SKBR-3 cell line. The most potent compound 13a induced the transcriptional activity of ER in dose-dependent manner in dual luciferase recombinant reporter model and increased progesterone receptor's expression, proving the retained estrogenic activity. The fluorescence of candidate compound 13a co-localised with the ERα. The newly synthesized labeled compounds might serve as good starting point for further development of fluorescent probes for modern biological applications. In addition to studying steroid uptake and transport in cells, e.g. in the processes of biodegradation of estrogen-hormones micropollutants, they could also be utilized in examination of estrogen-binding proteins.
- Klíčová slova
- 17β-estradiol, BODIPY, CuAAC, Estrogenic activity, Human estrogen receptor alpha,
- MeSH
- alfa receptor estrogenů * metabolismus chemie MeSH
- azidy chemie MeSH
- estradiol * chemie farmakologie MeSH
- estrogeny chemie MeSH
- fluorescenční barviva chemie MeSH
- lidé MeSH
- MFC-7 buňky MeSH
- nádorové buněčné linie MeSH
- proliferace buněk účinky léků MeSH
- simulace molekulového dockingu * MeSH
- sloučeniny boru * chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene MeSH Prohlížeč
- alfa receptor estrogenů * MeSH
- azidy MeSH
- ESR1 protein, human MeSH Prohlížeč
- estradiol * MeSH
- estrogeny MeSH
- fluorescenční barviva MeSH
- sloučeniny boru * MeSH
Thanks to their ability to bind to specific biological receptors, mannosylated structures are examined in biomedical applications. One of the most common ways of linking a functional moiety to a structure is to use an azide-alkyne click reaction. Therefore, it is necessary to prepare and isolate a propargylated mannose derivative of high purity to maintain its bioactivity. Three known preparations of propargyl-α-mannopyranoside were revisited, and products were analysed by NMR spectroscopy. The preparations were shown to yield by-products that have not been described in the literature yet. Our experiments showed that one-step procedures could not provide pure propargyl-α-mannopyranoside, while a three-step procedure yielded the desired compound of high purity.
- Klíčová slova
- NMR spectroscopy, alkylation, furanose, mannose, propargyl, pyranose,
- MeSH
- alkyny * chemie MeSH
- azidy chemie MeSH
- click chemie * metody MeSH
- magnetická rezonanční spektroskopie MeSH
- mannosa * chemie MeSH
- molekulární struktura MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- alkyny * MeSH
- azidy MeSH
- mannosa * MeSH
In continuation of our efforts to synthesize a highly dedicated strong cation exchanger, we introduce four chiral stationary phases based on a laterally substituted naphthalene core featuring chiral 2-aminocyclohexansulfonic acid as the chiral cation-exchange site. The selectors were modified with two different terminal units, which enabled immobilization to the silica support by thiol-ene radical reaction or azide-yne click chemistry. The chromatographic parameters of these chiral stationary phases were determined using a set of chiral amines, mainly from the family of β-blocker pharmaceuticals. The chiral stationary phases immobilized by means of click chemistry were found to be superior to those possessing the sulfide linker to the silica support. The chromatographic results and visualization of density functional theory-calculated conformations of the selectors hint at a combination of a steric and electronic effect of the triazole ring in the course of chiral resolution of the target analytes.
- Klíčová slova
- basic pharmaceuticals, chiral cation exchanger, chiral separation, chiral stationary phase, liquid chromatography,
- MeSH
- azidy chemie MeSH
- click chemie metody MeSH
- kationtoměniče chemie MeSH
- léčivé přípravky * analýza chemie izolace a purifikace MeSH
- molekulární modely MeSH
- naftaleny chemie MeSH
- stereoizomerie MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- azidy MeSH
- kationtoměniče MeSH
- léčivé přípravky * MeSH
- naftaleny MeSH
- naphthalene MeSH Prohlížeč
Antifouling polymer layers containing extracellular matrix-derived peptide motifs offer promising new options for biomimetic surface engineering. In this contribution, we report the design of antifouling vascular grafts bearing biofunctional peptide motifs for tissue regeneration applications based on hierarchical polymer brushes. Hierarchical diblock poly(methyl ether oligo(ethylene glycol) methacrylate-block-glycidyl methacrylate) brushes bearing azide groups (poly(MeOEGMA-block-GMA-N3)) were grown by surface-initiated atom transfer radical polymerization (SI-ATRP) and functionalized with biomimetic RGD peptide sequences. Varying the conditions of copper-catalyzed alkyne-azide "click" reaction allowed for the immobilization of RGD peptides in a wide surface concentration range. The synthesized hierarchical polymer brushes bearing peptide motifs were characterized in detail using various surface sensitive physicochemical methods. The hierarchical brushes presenting the RGD sequences provided excellent cell adhesion properties and at the same time remained resistant to fouling from blood plasma. The synthesis of anti-fouling hierarchical brushes bearing 1.2 × 103 nmol/cm2 RGD biomimetic sequences has been adapted for the surface modification of commercially available grafts of woven polyethylene terephthalate (PET) fibers. The fiber mesh was endowed with polymerization initiator groups via aminolysis and acylation reactions optimized for the material. The obtained bioactive antifouling vascular grafts promoted the specific adhesion and growth of endothelial cells, thus providing a potential avenue for endothelialization of artificial conduits.
- Klíčová slova
- RGD peptide, X-ray photoelectron spectroscopy, biomimetic surface, hierarchical bioactive polymer brushes, vascular graft, “click”-chemistry,
- MeSH
- adsorpce MeSH
- aminokyselinové motivy MeSH
- azidy chemie MeSH
- biokompatibilní potahované materiály * MeSH
- biomimetické materiály * MeSH
- buněčná adheze MeSH
- buněčné dělení MeSH
- cévní endotel fyziologie MeSH
- cévní protézy * MeSH
- click chemie MeSH
- endoteliální buňky pupečníkové žíly (lidské) MeSH
- imobilizované proteiny MeSH
- křemík MeSH
- krevní plazma MeSH
- krevní proteiny MeSH
- lidé MeSH
- oligopeptidy chemie MeSH
- polyethylentereftaláty chemie MeSH
- polymerizace * MeSH
- povrchové vlastnosti MeSH
- řízená tkáňová regenerace přístrojové vybavení MeSH
- sklo MeSH
- testování materiálů MeSH
- trombóza prevence a kontrola MeSH
- zlato MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Názvy látek
- arginyl-glycyl-aspartic acid MeSH Prohlížeč
- azidy MeSH
- biokompatibilní potahované materiály * MeSH
- imobilizované proteiny MeSH
- křemík MeSH
- krevní proteiny MeSH
- oligopeptidy MeSH
- polyethylentereftaláty MeSH
- zlato MeSH
To tailor cell-surface interactions, precise and controlled attachment of cell-adhesive motifs is required, while any background non-specific cell and protein adhesion has to be blocked effectively. Herein, a versatile and highly reproducible antifouling surface modification based on "clickable" groups and hierarchically structured diblock copolymer brushes for the controlled attachment of cells is reported. The polymer brush architecture combines an antifouling bottom block of poly(2-hydroxyethyl methacrylate) poly(HEMA) and an ultrathin azide-bearing top block, which can participate in well-established "click" reactions including the highly selective copper-catalyzed alkyne-azide cycloaddition (CuAAC) reaction under mild conditions. This straightforward approach allows the rapid conjugation of a cell-adhesive, alkyne-bearing cyclic RGD peptide motif, enabling subsequent specific attachment of NIH 3T3 fibroblasts, their extensive proliferation and confluent cell sheet formation after 48 h of incubation. The generally applicable strategy presented in this report can be employed for surface functionalization with diverse alkyne-bearing biological moieties via CuAAC or copper-free alkyne-azide cycloaddition protocols, making it a versatile functionalization approach and a promising tool for tissue engineering, biomaterial implant design, and other applications that require surfaces supporting highly specific cell attachment.
- Klíčová slova
- antifouling, controlled-cell attachment, copolymer brushes, surface biofunctionalization,
- MeSH
- alkyny chemie farmakologie MeSH
- antiinfekční látky chemická syntéza farmakologie MeSH
- azidy chemie farmakologie MeSH
- biokompatibilní materiály chemická syntéza farmakologie MeSH
- buňky NIH 3T3 MeSH
- click chemie MeSH
- cykloadiční reakce MeSH
- katalýza MeSH
- myši MeSH
- oligopeptidy chemie MeSH
- polyhydroxyethylmethakrylát chemie MeSH
- proliferace buněk účinky léků MeSH
- tkáňové inženýrství MeSH
- tkáňové podpůrné struktury * MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- alkyny MeSH
- antiinfekční látky MeSH
- arginyl-glycyl-aspartic acid MeSH Prohlížeč
- azidy MeSH
- biokompatibilní materiály MeSH
- oligopeptidy MeSH
- polyhydroxyethylmethakrylát MeSH
Though tetrapyrazinoporphyrazines (TPyzPzs) are generally presented as universal dark quenchers for oligonucleotide probes, the availability of TPyzPzs bearing different functional groups suitable for attachment to 3', and 5' ends or intrastrand positions remains rather limited. Therefore, a synthetic route to hexa(bis(2-methoxyethyl)amino) or hexa(diethylamino) TPyzPzs functionalized by an azide, hydroxy, or carboxy group or their combinations was developed. Studies of self-assembly into J-dimers in nonpolar solvents and their stability upon titration with pyridine (association constants, KP values, ranging 0.32-12.7×102 M-1 ) revealed that smaller peripheral substituents and functionalization of TPyzPzs improves the stability of J-dimers. ΦΔ and ΦF were low for the monomers (ΦF <0.0001, ΦΔ <0.008, DMF) due to quenching by intramolecular charge transfer; however, they increased in nonpolar solvents and after self-assembly into J-dimer (up to ΦF =0.027, ΦΔ =0.28).
- Klíčová slova
- aggregation, fluorescence, oligonucleotide probes, phthalocyanines, self-assembly,
- MeSH
- azidy chemie MeSH
- biosenzitivní techniky MeSH
- dimerizace MeSH
- fluorescenční barviva chemie MeSH
- fluorescenční spektrometrie MeSH
- komplexní sloučeniny chemie MeSH
- ligandy MeSH
- molekulární konformace MeSH
- porfyriny chemická syntéza MeSH
- pyridiny chemie MeSH
- rozpouštědla chemie MeSH
- rozpustnost MeSH
- termodynamika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- azidy MeSH
- fluorescenční barviva MeSH
- komplexní sloučeniny MeSH
- ligandy MeSH
- porfyriny MeSH
- pyridine MeSH Prohlížeč
- pyridiny MeSH
- rozpouštědla MeSH
Cyclic pentapeptides containing the amino acid sequence arginine-glycine-aspartic (RGD) have been widely applied to target αvβ3 integrin, which is upregulated in various tumors during tumor-induced angiogenesis. Multimeric cyclic RGD peptides have been reported to be advantageous over monomeric counterparts for angiogenesis imaging. Here, we prepared mono-, di-, and trimeric cyclic arginine-glycine-aspartic-D-phenylalanine-lysine (c (RGDfK)) derivatives by conjugation with the natural chelator fusarinine C (FSC) using click chemistry based on copper (I)-catalyzed azide-alkyne cycloaddition (CuAAC). The αvβ3 binding properties of 68Ga-labeled mono-, di-, and trimeric c(RGDfK) peptides were evaluated in vitro as well as in vivo and compared with the references monomeric [68Ga]GaNODAGA-c(RGDfK) and trimeric [68Ga]GaFSC(suc-c(RGDfK))3. All 68Ga-labeled c(RGDfK) peptides displayed hydrophilicity (logD = -2.96 to -3.80), low protein binding and were stable in phosphate buffered-saline (PBS) and serum up to 2 h. In vitro internalization assays with human melanoma M21 (αvβ3-positive) and M21-L (αvβ3-negative) cell lines showed specific uptake of all derivatives and increased in the series: mono- < di- < trimeric peptide. The highest tumor uptake, tumor-to-background ratios, and image contrast were found for the dimeric [68Ga]GaMAFC(c(RGDfK)aza)2. In conclusion, we developed a novel strategy for direct, straight forward preparation of mono-, di-, and trimeric c(RGDfK) conjugates based on the FSC scaffold. Interestingly, the best αvβ3 imaging properties were found for the dimeric [68Ga]GaMAFC(c(RGDfK)aza)2.
- Klíčová slova
- Angiogenesis, Gallium-68, PET, RGD, α(v)β(3) integrin,
- MeSH
- alkyny chemie MeSH
- azidy chemie MeSH
- click chemie MeSH
- cyklické peptidy chemie farmakokinetika MeSH
- izotopové značení MeSH
- měď chemie MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- PET/CT MeSH
- polymerizace MeSH
- radioizotopy galia chemie MeSH
- siderofory chemie MeSH
- tkáňová distribuce MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- alkyny MeSH
- azidy MeSH
- cyclic arginine-glycine-aspartic acid peptide MeSH Prohlížeč
- cyklické peptidy MeSH
- Gallium-68 MeSH Prohlížeč
- měď MeSH
- radioizotopy galia MeSH
- siderofory MeSH
The increasing popularity of peptides as promising molecular scaffolds for biomedical applications and as valuable biochemical probes makes new methods allowing for their modification highly desirable. We describe herein an optimized protocol based on a sequence of CuAAC click reactions and selective deprotection steps, which leads to an efficient multi-functionalization of synthetic peptides. The methodology has been successfully applied to the construction of defined heteroglycopeptides and fluorophore-quencher-containing probes for proteases. The developed chemistry thus represents an important addition to the available toolbox of methods enabling efficient postsynthetic modification of peptides. The commercial availability of numerous azide probes further greatly extends the application potential of the described methodology.
Copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) click reaction in the major groove of DNA containing 5-ethynyluracil (UE ) with azides was used for turning off sequence-specific protein-DNA interactions. The concept was first demonstrated on switching off cleavage of short modified DNA by restriction endonuclease BamHI-HF. Finally, DNA template containing UE was used for in vitro transcription with E. coli RNA polymerase and the transcription was turned off by CuAAC with 3-azidopropane-1,2-diol or 3-azido-7-hydroxycoumarin.
- Klíčová slova
- DNA, RNA polymerase, epigenetics, nucleotides, transcription,
- MeSH
- alkyny chemie MeSH
- azidy chemie MeSH
- bakteriální RNA chemie metabolismus MeSH
- click chemie MeSH
- cykloadiční reakce MeSH
- DNA řízené RNA-polymerasy chemie metabolismus MeSH
- DNA chemie MeSH
- Escherichia coli chemie MeSH
- katalýza MeSH
- kumariny chemie MeSH
- měď MeSH
- uracil analogy a deriváty MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- 3-azido-7-hydroxycoumarin MeSH Prohlížeč
- alkyny MeSH
- azidy MeSH
- bakteriální RNA MeSH
- DNA řízené RNA-polymerasy MeSH
- DNA MeSH
- eniluracil MeSH Prohlížeč
- kumariny MeSH
- měď MeSH
- uracil MeSH
The rise of CuI-catalyzed click chemistry has initiated an increased demand for azido and alkyne derivatives of amino acid as precursors for the synthesis of clicked peptides. However, the use of azido and alkyne amino acids in peptide chemistry is complicated by their high cost. For this reason, we investigated the possibility of the in-house preparation of a set of five Fmoc azido amino acids: β-azido l-alanine and d-alanine, γ-azido l-homoalanine, δ-azido l-ornithine and ω-azido l-lysine. We investigated several reaction pathways described in the literature, suggested several improvements and proposed several alternative routes for the synthesis of these compounds in high purity. Here, we demonstrate that multigram quantities of these Fmoc azido amino acids can be prepared within a week or two and at user-friendly costs. We also incorporated these azido amino acids into several model tripeptides, and we observed the formation of a new elimination product of the azido moiety upon conditions of prolonged couplings with 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate/DIPEA. We hope that our detailed synthetic protocols will inspire some peptide chemists to prepare these Fmoc azido acids in their laboratories and will assist them in avoiding the too extensive costs of azidopeptide syntheses. Experimental procedures and/or analytical data for compounds 3-5, 20, 25, 26, 30 and 43-47 are provided in the supporting information. © 2017 The Authors Journal of Peptide Science published by European Peptide Society and John Wiley & Sons Ltd.
- Klíčová slova
- alanine, azide elimination, azido amino acid, homoalanine, lysine, ornithine, synthesis,
- MeSH
- alkyny chemie MeSH
- aminokyseliny chemická syntéza MeSH
- azidy chemie MeSH
- click chemie metody MeSH
- ethylaminy chemie MeSH
- fluoreny chemická syntéza chemie MeSH
- močovina analogy a deriváty chemie MeSH
- peptidy chemická syntéza MeSH
- triazoly chemie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate MeSH Prohlížeč
- alkyny MeSH
- aminokyseliny MeSH
- azidy MeSH
- ethylaminy MeSH
- fluoreny MeSH
- močovina MeSH
- N,N-diisopropylethylamine MeSH Prohlížeč
- N(alpha)-fluorenylmethyloxycarbonylamino acids MeSH Prohlížeč
- peptidy MeSH
- triazoly MeSH
