protein mimetics
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Multi-orthogonal molecular scaffolds can be applied as core structures of bioactive compounds. Here, we prepared four tri-orthogonal scaffolds based on adamantane or proline skeletons. The scaffolds were used for the solid-phase synthesis of model insulin mimetics bearing two different peptides on the scaffolds. We found that adamantane-derived compounds bind to the insulin receptor more effectively (Kd value of 0.5 μM) than proline-derived compounds (Kd values of 15-38 μM) bearing the same peptides. Molecular dynamics simulations suggest that spacers between peptides and central scaffolds can provide greater flexibility that can contribute to increased binding affinity. Molecular modeling showed possible binding modes of mimetics to the insulin receptor. Our data show that the structure of the central scaffold and flexibility of attached peptides in this type of compound are important and that different scaffolds should be considered when designing peptide hormone mimetics.
- MeSH
- adamantan chemie MeSH
- inzulin analogy a deriváty chemická syntéza metabolismus MeSH
- kinetika MeSH
- krysa rodu rattus MeSH
- kvarterní struktura proteinů MeSH
- lidé MeSH
- prolin chemie MeSH
- receptor inzulinu chemie metabolismus MeSH
- simulace molekulární dynamiky MeSH
- stabilita proteinů MeSH
- stereoizomerie MeSH
- techniky syntézy na pevné fázi MeSH
- vazba proteinů MeSH
- vazebná místa MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Hematologic malignancies (HM) comprise diverse cancers of lymphoid and myeloid origin, including lymphomas (approx. 40%), chronic lymphocytic leukemia (CLL, approx. 15%), multiple myeloma (MM, approx. 15%), acute myeloid leukemia (AML, approx. 10%), and many other diseases. Despite considerable improvement in treatment options and survival parameters in the new millennium, many patients with HM still develop chemotherapy‑refractory diseases and require re-treatment. Because frontline therapies for the majority of HM (except for CLL) are still largely based on classical cytostatics, the relapses are often associated with defects in DNA damage response (DDR) pathways and anti-apoptotic blocks exemplified, respectively, by mutations or deletion of the TP53 tumor suppressor, and overexpression of anti-apoptotic proteins of the B-cell lymphoma 2 (BCL2) family. BCL2 homology 3 (BH3) mimetics represent a novel class of pro-apoptotic anti-cancer agents with a unique mode of action-direct targeting of mitochondria independently of TP53 gene aberrations. Consequently, BH3 mimetics can effectively eliminate even non-dividing malignant cells with adverse molecular cytogenetic alterations. Venetoclax, the nanomolar inhibitor of BCL2 anti-apoptotic protein has been approved for the therapy of CLL and AML. Numerous venetoclax-based combinatorial treatment regimens, next-generation BCL2 inhibitors, and myeloid cell leukemia 1 (MCL1) protein inhibitors, which are another class of BH3 mimetics with promising preclinical results, are currently being tested in several clinical trials in patients with diverse HM. These pivotal trials will soon answer critical questions and concerns about these innovative agents regarding not only their anti-tumor efficacy but also potential side effects, recommended dosages, and the optimal length of therapy as well as identification of reliable biomarkers of sensitivity or resistance. Effective harnessing of the full therapeutic potential of BH3 mimetics is a critical mission as it may directly translate into better management of the aggressive forms of HM and could lead to significantly improved survival parameters and quality of life in patients with urgent medical needs.
- MeSH
- apoptóza fyziologie MeSH
- bicyklické sloučeniny heterocyklické terapeutické užití MeSH
- biologické markery metabolismus MeSH
- hematologické nádory farmakoterapie metabolismus MeSH
- lidé MeSH
- nádorový supresorový protein p53 metabolismus MeSH
- poškození DNA účinky léků genetika MeSH
- sulfonamidy terapeutické užití MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Caloric restriction mimetics (CRMs) mimic the biochemical effects of nutrient deprivation by reducing lysine acetylation of cellular proteins, thus triggering autophagy. Treatment with the CRM hydroxycitrate, an inhibitor of ATP citrate lyase, induced the depletion of regulatory T cells (which dampen anticancer immunity) from autophagy-competent, but not autophagy-deficient, mutant KRAS-induced lung cancers in mice, thereby improving anticancer immunosurveillance and reducing tumor mass. Short-term fasting or treatment with several chemically unrelated autophagy-inducing CRMs, including hydroxycitrate and spermidine, improved the inhibition of tumor growth by chemotherapy in vivo. This effect was only observed for autophagy-competent tumors, depended on the presence of T lymphocytes, and was accompanied by the depletion of regulatory T cells from the tumor bed.
- MeSH
- Atg5 genetika MeSH
- autofagie MeSH
- citráty aplikace a dávkování farmakologie MeSH
- experimentální nádory dietoterapie farmakoterapie imunologie MeSH
- kalorická restrikce metody MeSH
- lidé MeSH
- methotrexát aplikace a dávkování farmakologie MeSH
- monitorování imunologické MeSH
- mutace MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- protoonkogenní proteiny p21(ras) genetika MeSH
- regulační T-lymfocyty účinky léků MeSH
- spermidin aplikace a dávkování farmakologie MeSH
- transplantace nádorů MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
Immune checkpoint blockade (ICB) using monoclonal antibodies against programmed cell death protein 1 (PD-1) or programmed death-ligand 1 (PD-L1) is the treatment of choice for cancer immunotherapy. However, low tissue permeability, immunogenicity, immune-related adverse effects, and high cost could be possibly improved using alternative approaches. On the other hand, synthetic low-molecular-weight (LMW) PD-1/PD-L1 blockers have failed to progress beyond in vitro studies, mostly due to low binding affinity or poor pharmacological characteristics resulting from their limited solubility and/or stability. Here, we report the development of polymer-based anti-human PD-L1 antibody mimetics (α-hPD-L1 iBodies) by attaching the macrocyclic peptide WL12 to a N-(2-hydroxypropyl)methacrylamide copolymer. We characterized the binding properties of iBodies using surface plasmon resonance, enzyme-linked immunosorbent assay, flow cytometry, confocal microscopy, and a cellular ICB model. We found that the α-hPD-L1 iBodies specifically target human PD-L1 (hPD-L1) and block the PD-1/PD-L1 interaction in vitro, comparable to the atezolizumab, durvalumab, and avelumab licensed monoclonal antibodies targeting PD-L1. Our findings suggest that iBodies can be used as experimental tools to target hPD-L1 and could serve as a platform to potentiate the therapeutic effect of hPD-L1-targeting small molecules by improving their affinity and pharmacokinetic properties.
Apoptóza (programovaná buněčná smrt) hraje významnou roli v patogenezi, progresi choroby a rezistenci k terapii u většiny nádorů krvetvorné a lymfatické tkáně. Vývoj a testování malých molekul blokují- cích antiapoptotické proteiny je jednou z možných a účinných cest, jak klinicky vý- znamně indukovat buněčnou smrt nádorových buněk, a tím léčebně ovlivnit průběh těchto chorob. Prvním účinným a klinicky dostupným představitelem nové třídy protinádorových léků zacílených na apoptózu, označovaných jako inhibitory proteinu BCL2, respektive BH3-mimetika, je venetoklax (ABT-199/GDC-0199). Ten prokázal v klinických studiích významný protinádorový účinek v monoterapii u refrakterních a relabovaných forem chronické lymfocytární leukemie (CLL), a to včetně nemocných s delecí krátkých ramen chromosomu 17 / mutacemi TP53 (nefunkční dráha proteinu p53) a pacientů s relapsem či refrakterních vůči léčbě ibrutinibem nebo idelalisibem. Vysoká účinnost venetoklaxu, jeho dobrá snášenlivost a přijatelný profil toxicity z něj činí nový významný nástroj v léčbě CLL, ale i dalších nádorových chorob krvetvorné a lymfatické tkáně. Jeho potenciál mohou do budoucna ještě výrazně zvýšit kombinace s jinými moderními protinádorovými léky, které jsou nyní testovány v řadě klinických studií.
Apoptosis (programmed cell death) plays an important role in the pathogenesis and progression of disease and in resistance to therapy in most haematopoietic and lymphatic tissue tumours. Development and testing of small molecules that block anti-apoptotic proteins is one of the possible and effective ways how to induce clinically significant cell death of tumour cells providing an instrument how to therapeutically influence the course of the disease. First effective and clinically available representative of a new class of anti-tumour drugs targeting apoptosis, referred to as BCL2 protein inhibitors, or BH3-mimetics, is venetoclax (ABT-199/GDC-0199). The agent has shown significant anti-tumour efficacy in clinical trials as monotherapy of refractory and relapsing forms of chronic lymphocytic leukaemia (CLL), including patients with deletions of the short arms of chromosome 17 / TP53 mutations (non-functional p53 protein pathway) and patients with a relapse or refractory to therapy with ibrutinib or idelalisib. The high efficacy of venetoclax, its good tolerance and acceptable toxicity profile make it a new important instrument in the treatment of CLL, but also of other haematopoietic and lymphatic tissue tumours. In the near future, its potential may still be markedly increased as part of combinations with other novel anti-tumour drugs that are now undergoing testing in a number of clinical trials.
- Klíčová slova
- Venclyxto - venetoklax tbl,
- MeSH
- apoptóza účinky léků MeSH
- bicyklické sloučeniny heterocyklické aplikace a dávkování farmakologie MeSH
- chronická lymfatická leukemie * farmakoterapie genetika MeSH
- farmakologické jevy MeSH
- hodnocení rizik MeSH
- lidé MeSH
- nežádoucí účinky léčiv MeSH
- protein BCL2L11 účinky léků MeSH
- protoonkogenní proteiny účinky léků MeSH
- rozvrh dávkování léků MeSH
- sulfonamidy aplikace a dávkování farmakologie MeSH
- syndrom nádorového rozpadu diagnóza epidemiologie farmakoterapie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- Check Tag
- lidé MeSH
Direct interactions between proteins are essential for the regulation of their functions in biological pathways. Targeting the complex network of protein-protein interactions (PPIs) has now been widely recognized as an attractive means to therapeutically intervene in disease states. Even though this is a challenging endeavor and PPIs have long been regarded as "undruggable" targets, the last two decades have seen an increasing number of successful examples of PPI modulators, resulting in growing interest in this field. PPI modulation requires novel approaches and the integrated efforts of multiple disciplines to be a fruitful strategy. This perspective focuses on the hub-protein 14-3-3, which has several hundred identified protein interaction partners, and is therefore involved in a wide range of cellular processes and diseases. Here, we aim to provide an integrated overview of the approaches explored for the modulation of 14-3-3 PPIs and review the examples resulting from these efforts in both inhibiting and stabilizing specific 14-3-3 protein complexes by small molecules, peptide mimetics, and natural products.
The search for potential inhibitors that target so far unexplored bacterial enzyme mono-N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE) has stimulated a development of methodology for quick and efficient preparation of mono-N-acylated 2,6-diaminopimelic acid (DAP) derivatives bearing the different carboxyl groups or lipophilic moieties on their amino group.
- MeSH
- acylace MeSH
- biomimetické materiály chemická syntéza chemie MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- kyselina diaminopimelová analogy a deriváty chemická syntéza chemie MeSH
- metabolické sítě a dráhy MeSH
- molekulární modely MeSH
- sukcináty chemická syntéza chemie MeSH
- sukcinyldiaminopimeláttransaminasa antagonisté a inhibitory metabolismus MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The polyhistidine tag (His-tag) is one of the most commonly used epitope tags in protein engineering. While His-tagged proteins can be detected reliably using immunological methods such as ELISA and Western blot, these methods are costly and time-intensive, necessitating more facile solutions for preliminary qualitative determination and concentration estimation. To this end, we present a rapid test strip assay based on iBody antibody mimetics that target the His-tag. We compare this strategy to commercial antibody-based assays and discuss the advantages and caveats of lateral flow assay design. Our test strip detected a panel of His-tagged proteins with different tag attachment strategies with a visual detection limit of 1 μM and densitometric detection limit of 0.5 μM. Due to its chemical nature, the presented assay exhibits wide reagent compatibility in comparison to antibody-based assays.
CHIP is a tetratricopeptide repeat (TPR) domain protein that functions as an E3-ubiquitin ligase. As well as linking the molecular chaperones to the ubiquitin proteasome system, CHIP also has a docking-dependent mode where it ubiquitinates native substrates, thereby regulating their steady state levels and/or function. Here we explore the effect of Hsp70 on the docking-dependent E3-ligase activity of CHIP. The TPR-domain is revealed as a binding site for allosteric modulators involved in determining CHIP's dynamic conformation and activity. Biochemical, biophysical and modeling evidence demonstrate that Hsp70-binding to the TPR, or Hsp70-mimetic mutations, regulate CHIP-mediated ubiquitination of p53 and IRF-1 through effects on U-box activity and substrate binding. HDX-MS was used to establish that conformational-inhibition-signals extended from the TPR-domain to the U-box. This underscores inter-domain allosteric regulation of CHIP by the core molecular chaperones. Defining the chaperone-associated TPR-domain of CHIP as a manager of inter-domain communication highlights the potential for scaffolding modules to regulate, as well as assemble, complexes that are fundamental to protein homeostatic control.
- MeSH
- alosterická regulace MeSH
- exprese genu MeSH
- interferonový regulační faktor 1 genetika metabolismus MeSH
- kinetika MeSH
- lidé MeSH
- lymfocyty cytologie metabolismus MeSH
- mapování interakce mezi proteiny MeSH
- molekulární modely MeSH
- nádorové buněčné linie MeSH
- nádorový supresorový protein p53 genetika metabolismus MeSH
- proteasomový endopeptidasový komplex metabolismus MeSH
- proteiny tepelného šoku HSP70 chemie genetika metabolismus MeSH
- sekundární struktura proteinů MeSH
- terciární struktura proteinů MeSH
- ubikvitinace MeSH
- ubikvitinligasy chemie genetika metabolismus MeSH
- vazba proteinů MeSH
- vazebná místa MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
