The worldwide COVID-19 pandemic caused by the coronavirus SARS-CoV-2 urgently demands novel direct antiviral treatments. The main protease (Mpro) and papain-like protease (PLpro) are attractive drug targets among coronaviruses due to their essential role in processing the polyproteins translated from the viral RNA. In this study, we virtually screened 688 naphthoquinoidal compounds and derivatives against Mpro of SARS-CoV-2. Twenty-four derivatives were selected and evaluated in biochemical assays against Mpro using a novel fluorogenic substrate. In parallel, these compounds were also assayed with SARS-CoV-2 PLpro. Four compounds inhibited Mpro with half-maximal inhibitory concentration (IC50) values between 0.41 μM and 9.0 μM. In addition, three compounds inhibited PLpro with IC50 ranging from 1.9 μM to 3.3 μM. To verify the specificity of Mpro and PLpro inhibitors, our experiments included an assessment of common causes of false positives such as aggregation, high compound fluorescence, and inhibition by enzyme oxidation. Altogether, we confirmed novel classes of specific Mpro and PLpro inhibitors. Molecular dynamics simulations suggest stable binding modes for Mpro inhibitors with frequent interactions with residues in the S1 and S2 pockets of the active site. For two PLpro inhibitors, interactions occur in the S3 and S4 pockets. In summary, our structure-based computational and biochemical approach identified novel naphthoquinonal scaffolds that can be further explored as SARS-CoV-2 antivirals.

• MeSH
• antivirové látky * farmakologie   chemie   MeSH
• COVID-19 MeSH
• inhibitory proteas * farmakologie   chemie   MeSH
• koronavirové proteasy 3C * antagonisté a inhibitory   MeSH
• koronavirové proteasy podobné papainu * antagonisté a inhibitory   MeSH
• lidé MeSH
• naftochinony * chemie   farmakologie   MeSH
• papain MeSH
• SARS-CoV-2 * účinky léků   enzymologie   MeSH
• simulace molekulového dockingu MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• 3C-like proteinase, SARS-CoV-2 MeSH Prohlížeč
• antivirové látky * MeSH
• inhibitory proteas * MeSH
• koronavirové proteasy 3C * MeSH
• koronavirové proteasy podobné papainu * MeSH
• naftochinony * MeSH
• papain-like protease, SARS-CoV-2 MeSH Prohlížeč
• papain MeSH

Adsorption properties of protein Papain at the solid|liquid (0.1 M KCl) interfaces of different hydrophobicity [highly oriented pyrolytic graphite (HOPG), bare gold, CH3, OH, and COOH-terminated self-assembled monolayers on gold] were studied by a combined quartz crystal microbalance and atomic force microscopy techniques. It was found that Papain forms an incomplete monolayer at hydrophobic interfaces (HOPG and CH3-terminated substrate), whereas on more hydrophilic ones, a complete monolayer formation was always observed with either the onset of the formation of a second layer (bare gold substrate) or adsorption in a multilayer fashion, possibly a bilayer formation (OH-terminated substrate). The surface concentration and compact monolayer film thickness was much lower on the COOH-terminated substrate compared to other surfaces studied. This result was explained by partial dissociation of the interfacial COOH groups leading to additional electrostatic interactions between the positively charged protein domains and negatively charged carboxylate anions, as well as to local pH changes promoting protein denaturation.

• MeSH
• adsorpce * MeSH
• grafit chemie   MeSH
• hydrofobní a hydrofilní interakce * MeSH
• mikrorovnovážné techniky křemenného krystalu MeSH
• mikroskopie atomárních sil MeSH
• papain metabolismus   MeSH
• povrchové vlastnosti * MeSH
• zlato chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• grafit MeSH
• papain MeSH
• zlato MeSH

Hyaluronic acid (HA) is known to serve as a dynamic mediator intervening in many physiological functions. Its specific effect has been repeatedly confirmed to be strongly influenced by the molecular size of hyaluronan fragments. However common technological approaches of HA fragments production have their limitations. In many cases, the final products do not meet the strict pharmaceutical requirements, specifically due to size polydispersity and reaction contaminants. We present novel methodology based on combination of unique incidental ability of the plant-derived protease papain to split the glycosidic bonds and an indispensable advantages of biocompatible macroporous material with incorporated ferrous ions serving as carrier for covalent papain fixation. This atypical and yet unpublished highly efficient multiparametric approach allows enhanced HA fragmentation for easily and safely producing molar-mass-defined HA fragments with narrow size distribution. Native polyacrylamide gel electrophoresis (PAGE) and size exclusion chromatography/multi-angle light scattering (SEC-MALS) confirmed the effectiveness of our multiparametric approach.

• Klíčová slova
• Hyaluronan fragmentation, Magnetic particles, Papain, Pharmaceutics, SEC-MALS,
• MeSH
• celulosa chemie   MeSH
• enzymy imobilizované chemie   MeSH
• gelová chromatografie MeSH
• hyaluronoglukosaminidasa chemie   MeSH
• kyselina hyaluronová chemie   MeSH
• magnetické jevy MeSH
• molekulová hmotnost MeSH
• nativní elektroforéza na polyakrylamidovém gelu MeSH
• papain chemie   MeSH
• radiační rozptyl MeSH
• viskozita MeSH
• železo chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• celulosa MeSH
• enzymy imobilizované MeSH
• hyaluronoglukosaminidasa MeSH
• kyselina hyaluronová MeSH
• papain MeSH
• železo MeSH

Nanobubbles formed on monocrystalline gold/water interface by means of the ethanol-to-water solvent exchange were exposed to the solutions of either bovine serum albumin or papain proteins. Both proteins do not change the position of nanobubbles in water, as observed by in situ tapping mode atomic force microscopy imaging before and after the introduction of the protein. The aqueous environment was subsequently replaced by ethanol. While all nanobubbles were found to dissolve in ethanol in the presence of bovine serum albumin, most of them survived when papain was employed. The protective ability of papain was ascribed to its resistance towards the protein denaturation in aqueous solutions of ethanol. The authors employed in situ atomic force nanolithography to investigate the nanomorphology of the papain/nanobubble assemblies in ethanol.

• MeSH
• ethanol chemie   MeSH
• nanostruktury chemie   MeSH
• papain metabolismus   MeSH
• povrchové vlastnosti MeSH
• sérový albumin hovězí metabolismus   MeSH
• skot MeSH
• voda chemie   MeSH
• zlato metabolismus   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ethanol MeSH
• papain MeSH
• sérový albumin hovězí MeSH
• voda MeSH
• zlato MeSH

To study molecular mechanisms underlying self-defense of the bacterial pathogen Plesiomonas shigelloides against host inflammatory and immune responses, we evaluated its interactions with mammalian papain-like cathepsins that are essential for host immunity. When grown under anaerobic, but not aerobic, conditions, P. shigelloides was shown to bind and inhibit papain, a model representative of the papain family of cysteine proteinases. This points to mammalian cathepsins as likely physiological targets of a novel cysteine-proteinase inhibitor expressed on bacterial cell surface. Both papain and mammalian cathepsins L and B were inhibited by periplasmic extracts of aerobically and anaerobically grown bacteria, the inhibitory activity being higher in the latter. Inhibition by both intact cells and periplasmic samples was rapid and efficient. The results suggest a possible defensive role of bacterial inhibitors of cathepsins during invasion of a mammalian host. The bacteria thus may modulate host protective responses through inhibiting cathepsins involved in antigen processing and presentation.

• MeSH
• antigeny bakteriální MeSH
• cysteinové endopeptidasy MeSH
• inhibitory cysteinových proteinas metabolismus   farmakologie   MeSH
• kathepsin B antagonisté a inhibitory   MeSH
• kathepsin L MeSH
• kathepsiny antagonisté a inhibitory   MeSH
• lidé MeSH
• papain antagonisté a inhibitory   MeSH
• periplazma metabolismus   MeSH
• Plesiomonas imunologie   metabolismus   patogenita   MeSH
• prezentace antigenu MeSH
• savci MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny bakteriální MeSH
• CTSL protein, human MeSH Prohlížeč
• cysteinové endopeptidasy MeSH
• inhibitory cysteinových proteinas MeSH
• kathepsin B MeSH
• kathepsin L MeSH
• kathepsiny MeSH
• papain MeSH

Using two different kinds of pH systems--the papain catalyzed hydrolysis of N-benzoyl-L-arginine ethyl ester in a membrane reactor and the bromate-sulfite-ferrocyanide (BSF) reaction in the CSTR--we study the relation among excitability, oscillations and bistability, and the ability of the system to respond to external periodic perturbations. Excitable properties of dynamical systems are examined in terms of a threshold set which is used to characterise dynamics in the reactor subject to external periodic stimuli. A precise definition and a method of calculating the threshold set are formulated. Two kinds of excitability distinguished by either direct or indirect initiation of the activatory process are found in both pH systems. Periodic pulsed perturbations of the BSF system display a nontrivial dependence of an excitation number on the forcing period. We examined this system also in oscillatory mode by looking at the phase shifts caused by single-pulse perturbations and constructing the phase transition curves (PTCs).

• MeSH
• algoritmy MeSH
• bromičnany chemie   MeSH
• ferrokyanidy chemie   MeSH
• katalýza * MeSH
• kinetika MeSH
• koncentrace vodíkových iontů MeSH
• membrány umělé MeSH
• nelineární dynamika MeSH
• papain chemie   MeSH
• siřičitany chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bromičnany MeSH
• ferrokyanidy MeSH
• hexacyanoferrate II MeSH Prohlížeč
• membrány umělé MeSH
• papain MeSH
• siřičitany MeSH

PURPOSE: The aim of the present study was to investigate the effect of a mixture of proteolytic enzymes (comprising trypsin, chymotrypsin and papain) on the metastatic model of syngeneic melanoma B16. METHODS: 140 C57B16 mice were divided into two control and two "treated" groups. Control groups received saline rectally, twice a day starting 24 h after intracutaneous transplantation (C1) or from the time point of the primary B16 melanoma extirpation (C2), respectively. "Treated" groups were rectally administered a mixture of 0.2 mg trypsin, 0.5 mg papain, and 0.2 mg chymotrypsin twice daily starting 24 h after transplantation (E1) or after extirpation of the tumor (E2), respectively. Survival of mice and B16 melanoma generalization were observed for a period of 100 days. Immunological evaluation of B16 melanoma cells in the ascites was accomplished. CD44, CD54 and CD106 cells were measured by flow cytometry. RESULTS: Administration of proteolytic enzymes to mice inhibited the growth of primary tumors, and tumor recurrences were less numerous. Importantly, metastasis was considerably curtailed both in the vicinity of the primary tumor and at distant locales. These findings correlated with a decreased expression of CD44 and CD54 molecules in tumors exposed to proteolytic enzymes in vivo. CONCLUSIONS: Our data suggest that serine and cysteine proteinases suppress B16 melanoma, and restrict its metastatic dissemination in C57B16 mice.

• MeSH
• antigeny CD44 imunologie   MeSH
• antigeny povrchové imunologie   MeSH
• aplikace rektální MeSH
• buněčné dělení účinky léků   MeSH
• chymotrypsin aplikace a dávkování   farmakologie   MeSH
• endopeptidasy farmakologie   MeSH
• fixní kombinace léků MeSH
• imunoglobuliny imunologie   MeSH
• melanom experimentální farmakoterapie   imunologie   patologie   sekundární   MeSH
• mezibuněčná adhezivní molekula-1 imunologie   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádory plic imunologie   prevence a kontrola   sekundární   MeSH
• papain aplikace a dávkování   farmakologie   MeSH
• protinádorové látky farmakologie   MeSH
• trypsin aplikace a dávkování   farmakologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigen 106 MeSH Prohlížeč
• antigeny CD44 MeSH
• antigeny povrchové MeSH
• chymotrypsin, papain, trypsin drug combination MeSH Prohlížeč
• chymotrypsin MeSH
• endopeptidasy MeSH
• fixní kombinace léků MeSH
• imunoglobuliny MeSH
• mezibuněčná adhezivní molekula-1 MeSH
• papain MeSH
• protinádorové látky MeSH
• trypsin MeSH

We studied the mode of regulation of the activity of mature cathepsin B (CB) by L-cysteine and some related thiols. The activity of CB with Z-Arg-Arg-NHMec as substrate was gradually inhibited over a range of increasing concentration of Cys, Cys methyl ester (CysOMe), Cys ethyl ester (CysOEt), N-acetyl-Cys (N-AcCys) and 3-mercaptopropionic acid. However, the inhibition of CB peaked at a definite value of [Cys], [CysOMe], [CysOEt] and [N-AcCys] and was gradually reversed over a range of higher concentrations of Cys and its esters. The maximum inhibitory concentrations of Cys, CysOME, CysOEt and N-AcCys showed a positive relationship to the pKa(RSH) values of the thiols and those of CysOEt and Cys decreased with increasing pH. The capability of the thiols to overcome their own inhibitory effect on CB was dependent on the concentration of their thiolate anion (RS-). However, the preincubation-dilution experiments showed that Cys and N-AcCys did not interact with active CB via a covalent mode. The inhibition of CB by N-AcCys was competitive and could be reversed by CysOMe. This activity-recovering effect of CysOMe was concentration-dependent and obeyed the Michaelis-Menten saturation kinetics over a profound increase of [RS-]. CB reacting in an environment of concurrently decreasing [RS-] and increasing [RSH], which was achieved by means of carboxylesterase-catalyzed deesterification of CysOEt to Cys, was progressively inhibited. Cys and N-AcCys also inhibited the fragmentation of histone H4 by CB and their concentration-dependent inhibitory profiles were qualitatively similar to those observed with Z-Arg-Arg-NHMec. Taken together, the results indicate that the RSH form of Cys and related thiols inhibits the activity of CB while the RS- form of these thiols counteracts or reverses the inhibitory action of the RSH form. This previously unrecognized thiol-thiolate anion regulation mechanism might be involved in a dynamic regulation of CB activity in endosomes and lysosomes and at the sites of lysosome-driven pericellular proteolysis.

• MeSH
• aktivace enzymů MeSH
• cystein farmakologie   MeSH
• hydrolýza MeSH
• játra účinky léků   enzymologie   MeSH
• kathepsin B antagonisté a inhibitory   metabolismus   MeSH
• kinetika MeSH
• koncentrace vodíkových iontů MeSH
• lidé MeSH
• papain antagonisté a inhibitory   MeSH
• substrátová specifita MeSH
• sulfhydrylové sloučeniny farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cystein MeSH
• kathepsin B MeSH
• papain MeSH
• sulfhydrylové sloučeniny MeSH

Long-term rectal administration of enzyme mixture containing papain, trypsin and chymotrypsin in the same ratio as the preparation Wobe-Mugos E (Mucos Pharma, Germany) was evaluated for their antitumor effects in C57Bl6 inbred mice inoculated with Bl6 melanoma cells. 30% of animals in the test group (3 pcs) have been cured of cancer. In the rest of animals (70%) the survival time was prolonged by 58.3% compared to the control group (from average survival time of 24 days in control group to 38 days in the test group). Based on histological and immunohistochemical evaluation a faster process of metastasizing was found in control group than in the group treated with the polyenzyme preparation. In the case of melanoma Bl6 an antimetastatic effect of the preparation was thus proved.

• MeSH
• buněčné dělení účinky léků   MeSH
• chymotrypsin * MeSH
• extrakty z thymu farmakologie   MeSH
• fixní kombinace léků MeSH
• imunohistochemie MeSH
• melanom experimentální metabolismus   patologie   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádory plic prevence a kontrola   sekundární   MeSH
• pankreatické extrakty farmakologie   MeSH
• papain farmakologie   MeSH
• trypsin * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chymotrypsin, papain, trypsin drug combination MeSH Prohlížeč
• chymotrypsin * MeSH
• extrakty z thymu MeSH
• fixní kombinace léků MeSH
• pankreatické extrakty MeSH
• papain MeSH
• trypsin * MeSH

The effect of combined proteolytic enzymes, administered by the rectal route, on the metastatic process and the time of survival in C57Bl6 mice with the Lewis lung carcinoma inoculated subcutaneously was investigated. In the control group, which received no enzyme treatment, 90% of animals died of the metastatic spread of cancer by day 18 after primary tumor extirpation. In Group A, which received the multi-enzyme solution from the time of primary tumor extirpation, 30% of mice died of disseminated cancer by day 25. In Group B, which was treated with the enzymes from 6 days before primary tumor extirpation, only 10% of animals showed the metastatic process by day 15. In Group C, which received the enzymes from 24 hours after intracutaneous tumor inoculation, no metastatic dissemination was discernible. In these three groups, the enzyme treatment was carried out throughout the study. None of the control animals survived for 100 days when the study was ended. The treated groups A, B and C showed survival rate 60%, 90% and 100% of animals, respectively, by 100 days.

• MeSH
• aplikace rektální MeSH
• chymotrypsin * MeSH
• endopeptidasy aplikace a dávkování   farmakologie   MeSH
• extrakty z thymu aplikace a dávkování   farmakologie   MeSH
• fixní kombinace léků MeSH
• karcinom plic Lewisové mortalita   patologie   prevence a kontrola   MeSH
• metastázy nádorů patologie   prevence a kontrola   MeSH
• míra přežití MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádory kůže mortalita   patologie   prevence a kontrola   MeSH
• pankreatické extrakty aplikace a dávkování   farmakologie   MeSH
• papain aplikace a dávkování   farmakologie   MeSH
• transplantace nádorů MeSH
• trypsin * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chymotrypsin, papain, trypsin drug combination MeSH Prohlížeč
• chymotrypsin * MeSH
• endopeptidasy MeSH
• extrakty z thymu MeSH
• fixní kombinace léků MeSH
• pankreatické extrakty MeSH
• papain MeSH
• trypsin * MeSH