The cellular adhesion receptor αvβ6-integrin is highly expressed in many cancers, e.g., pancreatic, lung, head-and-neck, cervical, bladder, and esophageal carcinoma. Multimerization of αvβ6-integrin-specific RGD peptides increases the target affinity and retention but affects biodistribution and pharmacokinetics. Amide formation of the terminal carboxylic acid moieties of the square-symmetrical bifunctional chelator DOTPI with 3-azidopropylamine yields derivatives with 4, 3, and 2 terminal azides and zero, 1, and 2 remaining carboxylic acids, respectively, whereby formation of the 2-cis-isomer is preferred according to NMR investigation of the Eu(III)-complexes. Cu(II)-catalyzed alkyne-azide cycloaddition (CuAAC) of the alkyne-functionalized αvβ6-integrin binding peptide cyclo[YRGDLAYp(NMe)K(pent-4-ynoic amide)] (Tyr2) yields the respective di-, tri-, and tetrameric conjugates for Lu-177-labeling. In mice bearing αvβ6-integrin-expressing xenografts of H2009 (human lung adenocarcinoma) cells, the Lu-177-labeled trimer's tumor-to-blood ratio of 112 exceeds that of the tetramer (10.4) and the dimer (54). Co-infusion of gelofusine (succinylated gelatin) reduces the renal uptake of the trimer by 89%, resulting in a 10-fold better tumor-to-kidney ratio, while no improvement of that ratio is observed with arginine/lysine, para-aminohippuric acid (PAH), and hydroxyethyl starch (HES) coinfusions. Since the Lu-177-labeled Tyr2-trimer outperforms the dimer and the tetramer, such trimers are considered the best lead structures for the ongoing development of αvβ6-integrin targeted anticancer theranostics.

• MeSH
• antigeny nádorové * metabolismus   MeSH
• chelátory * chemie   MeSH
• click chemie MeSH
• integriny * metabolismus   MeSH
• lidé MeSH
• lutecium * chemie   MeSH
• myši nahé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory farmakoterapie   MeSH
• oligopeptidy * chemie   farmakokinetika   MeSH
• radiofarmaka farmakokinetika   chemie   terapeutické užití   MeSH
• radionuklidy * chemie   MeSH
• teranostická nanomedicína metody   MeSH
• tkáňová distribuce MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Chemické transformace kompatibilní s biologickými systémy slouží jako neocenitelné nástroje pro zkoumání biomolekul, sloučenin léčiv a biologických procesů v jejich přirozeném prostředí. Složité prostředí živých organismů vyžaduje, aby tyto reakce byly vysoce selektivní a účinné, což představuje pro oblast organické chemie obrovskou výzvu. V poslední době se objevila řada chemických reakcí, které tyto podmínky splňují, a poskytují proto výzkumným pracovníkům rozmanitou sadu nástrojů. Tento rukopis představuje komplexní přehled současných bioortogonálních reakcí s důrazem na jejich aplikace v zobrazování, diagnostice a medicíně.

Chemical transformations compatible with biological systems serve as invaluable tools for probing biomolecules, drug compounds, and biological processes in their native environments. The complex environment of living organisms requires these reactions to be highly selective and efficient, posing a formidable challenge to the field of organic chemistry. A number of chemical reactions have recently emerged to meet this challenge, providing a diverse toolkit for researchers. This manuscript presents a comprehensive survey of current bioorthogonal reactions, emphasizing their applications in bioimaging, diagnostics, and medicine.

• MeSH
• click chemie * metody   MeSH
• fluorescenční barviva chemie   MeSH
• lékové transportní systémy metody   MeSH
• lidé MeSH
• molekulární zobrazování MeSH
• optické zobrazování metody   MeSH
• prekurzory léčiv farmakologie   MeSH
• Check Tag
• lidé MeSH

• Klíčová slova
• bioortogonální chemie,
• MeSH
• alkyny chemie   MeSH
• azidy chemie   MeSH
• chemické jevy MeSH
• click chemie * metody   MeSH
• cykloadiční reakce klasifikace   metody   přístrojové vybavení   MeSH
• výzkum MeSH
• Publikační typ
• přehledy MeSH

Bioorthogonal chemistry provides one of the possibilities to modify various biomolecules in their native environment. The combination of Click chemistry with the BONCAT method (bioorthogonal non-canonical amino acid tagging) is widely used for tagging and analysis of newly synthesized proteins, which are clearly distinguishable from the pre-existing protein pool. However, the commonly used procedure results in low quality 2D electrophoretic profiles. We put a lot of effort into obtaining clear results using a standard Click protocol, with a negligible effect. Here we describe a Click-on-membrane approach which we successfully used not only to monitor de novo protein synthesis but also to detect newly synthesized RNA.

• MeSH
• click chemie metody   MeSH
• proteiny analýza   MeSH
• RNA analýza   MeSH
• techniky kombinatorické chemie * metody   MeSH

Prostate cancer is primarily fatal after it becomes metastatic and castration-resistant despite novel combined hormonal and chemotherapeutic regimens. Hence, new therapeutic concepts and drug delivery strategies are urgently needed for the eradication of this devastating disease. Here we report the highly specific, in situ click chemistry driven pretargeted delivery of cytotoxic drug carriers to PSMA(+) prostate cancer cells. Anti-PSMA 5D3 mAb and its F(ab')2 fragments were functionalized with trans-cyclooctene (TCO), labeled with a fluorophore, and used as pretargeting components. Human serum albumin (ALB) was loaded with the DM1 antitubulin agent, functionalized with PEGylated tetrazine (PEG4-Tz), labeled with a fluorophore, and used as the drug delivery component. The internalization kinetics of components and the therapeutic efficacy of the pretargeted click therapy were studied in PSMA(+) PC3-PIP and PSMA(-) PC3-Flu control cells. The F(ab')2 fragments were internalized faster than 5D3 mAb in PSMA(+) PC3-PIP cells. In the two-component pretargeted imaging study, both components were colocalized in a perinuclear location of the cytoplasm of PC3-PIP cells. Better colocalization was achieved when 5D3 mAb was used as the pretargeting component. Consecutively, the in vitro cell viability study shows a significantly higher therapeutic effect of click therapy in PC3-PIP cells when 5D3 mAb was used for pretargeting, compared to its F(ab')2 derivative. 5D3 mAb has a longer lifetime on the cell surface, when compared to its F(ab')2 analogue, enabling efficient cross-linking with the drug delivery component and increased efficacy. Pretargeting and drug delivery components were cross-linked via multiple bioorthogonal click chemistry reactions on the surface of PSMA(+) PC cells forming nanoclusters, which undergo fast cellular internalization and intracellular transport to perinuclear locations.

• MeSH
• albuminy MeSH
• antigeny povrchové imunologie   MeSH
• click chemie metody   MeSH
• cyklooktany chemie   MeSH
• fluorbenzeny chemie   MeSH
• fytogenní protinádorové látky terapeutické užití   MeSH
• glutamátkarboxypeptidasa II imunologie   metabolismus   MeSH
• imunoglobuliny - Fab fragmenty chemie   metabolismus   terapeutické užití   MeSH
• lékové transportní systémy metody   MeSH
• lidé MeSH
• maytansin terapeutické užití   MeSH
• modulátory tubulinu terapeutické užití   MeSH
• monoklonální protilátky chemie   metabolismus   terapeutické užití   MeSH
• nádorové buněčné linie MeSH
• nádory prostaty farmakoterapie   enzymologie   metabolismus   MeSH
• nanomedicína MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH

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.

• 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

Hybrid imaging combining the beneficial properties of radioactivity and optical imaging within one imaging probe has gained increasing interest in radiopharmaceutical research. In this study, we modified the macrocyclic gallium-68 chelator fusarinine C (FSC) by conjugating a fluorescent moiety and tetrazine (Tz) moieties. The resulting hybrid imaging agents were used for pretargeting applications utilizing click reactions with a trans-cyclooctene (TCO) tagged targeting vector for a proof of principle both in vitro and in vivo. Starting from FSC, the fluorophores Sulfocyanine-5, Sulfocyanine-7, or IRDye800CW were conjugated, followed by introduction of one or two Tz motifs, resulting in mono and dimeric Tz conjugates. Evaluation included fluorescence microscopy, binding studies, logD, protein binding, in vivo biodistribution, µPET (micro-positron emission tomography), and optical imaging (OI) studies. 68Ga-labeled conjugates showed suitable hydrophilicity, high stability, and specific targeting properties towards Rituximab-TCO pre-treated CD20 expressing Raji cells. Biodistribution studies showed fast clearance and low accumulation in non-targeted organs for both SulfoCy5- and IRDye800CW-conjugates. In an alendronate-TCO based bone targeting model the dimeric IRDye800CW-conjugate resulted in specific targeting using PET and OI, superior to the monomer. This proof of concept study showed that the preparation of FSC-Tz hybrid imaging agents for pretargeting applications is feasible, making such compounds suitable for hybrid imaging applications.

• MeSH
• click chemie MeSH
• fluorescenční protilátková technika MeSH
• kyseliny hydroxamové * chemie   MeSH
• multimodální zobrazování * metody   MeSH
• optické zobrazování metody   MeSH
• ověření koncepční studie MeSH
• pozitronová emisní tomografie MeSH
• radiofarmaka * chemie   MeSH
• radioizotopy galia MeSH
• radionuklidy MeSH
• tkáňová distribuce MeSH
• železité sloučeniny * chemie   MeSH
• Publikační typ
• časopisecké články 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.

• 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

The site-specific chemical modification of proteins through incorporation of noncanonical amino acids enables diverse applications, such as imaging, probing, and expanding protein functions, as well as to precisely engineer therapeutics. Here we report a general strategy that allows the incorporation of noncanonical amino acids into target proteins using the amber suppression method and their efficient secretion in the biotechnological relevant expression host Bacillus subtilis. This facilitates efficient purification of target proteins directly from the supernatant, followed by their functionalization using click chemistry. We used this strategy to site-specifically introduce norbornene lysine into a single chain antibody and functionalize it with fluorophores for the detection of human target proteins.

• MeSH
• Bacillus subtilis genetika   metabolismus   MeSH
• click chemie MeSH
• CRISPR-Cas systémy MeSH
• ELISA MeSH
• genetické vektory MeSH
• genetický kód MeSH
• isopropylthiogalaktosid farmakologie   MeSH
• kreatinkinasa, forma MM metabolismus   MeSH
• lidé MeSH
• lysin chemie   MeSH
• norbornany chemie   MeSH
• proteinové inženýrství metody   MeSH
• regulace genové exprese u bakterií účinky léků   MeSH
• rekombinantní proteiny chemie   genetika   izolace a purifikace   metabolismus   MeSH
• zelené fluorescenční proteiny genetika   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In our work, we developed the synthesis of new polyfunctional pegylated trehalose derivatives and evaluated their cryoprotective effect using flow cytometry. We showed that new compounds (modified trehaloses) bound to appropriate extracellular polymeric cryoprotectants could be helpful as a chemical tool for the evaluation of their potential toxic cell membrane influences. Our aim was to form a chemical tool for the evaluation of cryoprotectant cell membrane influences, which are still not easily predicted during the freezing/thawing process. We combined two basic cryoprotectants: polyethyleneglycols (PEGs) and trehalose in the new chemical compounds-pegylated trehalose hybrids. If PEG and trehalose are chemically bound and trehalose is adsorbed on the cell surface PEGs molecules which are, due to the chemical bonding with trehalose, close to the cell surface, can remove the cell surface hydration layer which destabilizes the cell membrane. This was confirmed by the comparison of new material, PEG, trehalose, and their mixture cryoprotective capabilities.

• MeSH
• buněčná membrána účinky léků   metabolismus   MeSH
• click chemie MeSH
• dimethylsulfoxid farmakologie   MeSH
• kryoprezervace MeSH
• kryoprotektivní látky farmakologie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• polyethylenglykoly chemie   farmakologie   MeSH
• průtoková cytometrie MeSH
• trehalosa analogy a deriváty   chemická syntéza   chemie   farmakologie   MeSH
• viabilita buněk účinky léků   MeSH
• zmrazování MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH