scholarly journals Synthesis of N-benzoyl Amino Esters and N-benzoyl Amino Acids and their Antifungal Activity

2021 ◽  
Vol 66 (1) ◽  
Author(s):  
Yureli Chiguils-Pérez ◽  
Alejandro Israel Rodríguez-Hurtado ◽  
Lemuel Pérez-Picaso ◽  
Roxana Martínez-Pascual ◽  
María de los Ángeles Martínez-Rivera ◽  
...  
Keyword(s):  
Amino Acids ◽  
Antifungal Activity ◽  
In Vitro Assays ◽  
Side Chain ◽  
Binding Affinities ◽  
Aromatic Rings ◽  
Amino Esters ◽  
Chitinase A ◽  
Docking Calculations

Abstract. A series of N-benzoyl amino esters and N-benzoyl amino acids were synthesized from commercially-available amino acids (Val, Ile, Leu, Ala, Phe, Trp) and were evaluated for their antifungal activity against two filamentous fungi, A. fumigatus and F. temperatum. According to the in vitro assays, five compounds (5-7, 10, 13) exhibited relevant antifungal activity against F. temperatum and two compounds (5 and 7) showed remarkable activity against both fungi strains. Some structure-activity relationships were established regarding the side chain at Ca and the type of substituents on the aromatic ring in the benzoyl moiety. Docking calculations were performed in order to predict binding affinities between compounds prepared herein and fungal chitinase, a potential target against fungi; interactions involving the aromatic rings, the influence on the number of methyl substituents, and configurations on the a-carbon have been analyzed.   Resumen. Una serie de derivados N-benzoilamino ésteres y N-benzoilaminoácidos, sintetizados a partir de aminoácidos disponibles comercialmente (Val, Ile, Leu, Ala, Phe, Trp), se evaluaron como agentes antifúngicos frente a dos hongos filamentosos, A. fumigatus y F. temperatum. De acuerdo con los ensayos in vitro, cinco compuestos (5-7, 10, 13) exhibieron una actividad relevante contra F. temperatum y dos derivados (5 y 7) mostraron una actividad notable contra ambas cepas. Algunas relaciones de estructura actividad permitieron observar el efecto de la cadena lateral del aminoácido, y de los sustituyentes del grupo benzoílo, en la actividad biológica. Se realizaron cálculos de acoplamiento molecular con el propósito de predecir afinidades de enlace entre los compuestos sintetizados y la enzima quitinasa, considerada un blanco molecular potencial. Se analizaron las interacciones que involucran anillos aromáticos, la influencia de los sustituyentes metilo, así como la configuración del Ca.

scholarly journals Respiratory-induced coenzyme Q biosynthesis is regulated by a phosphorylation cycle of Cat5p/Coq7p

2011 ◽  
Vol 440 (1) ◽  
pp. 107-114 ◽  
Author(s):  
Alejandro Martín-Montalvo ◽  
Isabel González-Mariscal ◽  
Sergio Padilla ◽  
Manuel Ballesteros ◽  
David L. Brautigan ◽  
...  
Keyword(s):  
Amino Acids ◽  
Protein Kinase ◽  
Regulatory Mechanism ◽  
Coenzyme Q ◽  
In Vitro Assays ◽  
Respiratory Metabolism ◽  
Total Absence ◽  
Kinase C ◽  
Pkc Protein Kinase C

CoQ6 (coenzyme Q6) biosynthesis in yeast is a well-regulated process that requires the final conversion of the late intermediate DMQ6 (demethoxy-CoQ6) into CoQ6 in order to support respiratory metabolism in yeast. The gene CAT5/COQ7 encodes the Cat5/Coq7 protein that catalyses the hydroxylation step of DMQ6 conversion into CoQ6. In the present study, we demonstrated that yeast Coq7 recombinant protein purified in bacteria can be phosphorylated in vitro using commercial PKA (protein kinase A) or PKC (protein kinase C) at the predicted amino acids Ser20, Ser28 and Thr32. The total absence of phosphorylation in a Coq7p version containing alanine instead of these phospho-amino acids, the high extent of phosphorylation produced and the saturated conditions maintained in the phosphorylation assay indicate that probably no other putative amino acids are phosphorylated in Coq7p. Results from in vitro assays have been corroborated using phosphorylation assays performed in purified mitochondria without external or commercial kinases. Coq7p remains phosphorylated in fermentative conditions and becomes dephosphorylated when respiratory metabolism is induced. The substitution of phosphorylated residues to alanine dramatically increases CoQ6 levels (256%). Conversely, substitution with negatively charged residues decreases CoQ6 content (57%). These modifications produced in Coq7p also alter the ratio between DMQ6 and CoQ6 itself, indicating that the Coq7p phosphorylation state is a regulatory mechanism for CoQ6 synthesis.

scholarly journals Evaluation of the antifungal activity by plant extracts against Colletotrichum gloeosporioides Penz

2008 ◽  
Vol 32 (2) ◽  
pp. 420-428 ◽  
Author(s):  
Polyanna Alves Silva ◽  
Denilson Ferreira Oliveira ◽  
Ney Robson Taironi do Prado ◽  
Douglas Antônio de Carvalho ◽  
Gilvane Aparecida de Carvalho
Keyword(s):  
Antifungal Activity ◽  
Freeze Drying ◽  
Colletotrichum Gloeosporioides ◽  
Plantago Lanceolata ◽  
Stevia Rebaudiana ◽  
Tween 80 ◽  
In Vitro Assays ◽  
Bioactive Substances ◽  
Stevia Rebaudiana Bertoni

Aiming to develop more efficient and environmental friendly methods than those available to control Colletotrichum gloeosporioides Penz, which causes blister spot in coffee trees, a search for plants able to produce substances active against such pathogen was carried out. Thus, extracts of 48 plant species, collected at Alto Rio Grande region, in Minas Gerais, were prepared and submitted to in vitro assays with that fungus. The best results were obtained with the extracts prepared from Digitalis lanata Ehrh, Origanum manjorona L., Plantago lanceolata Hook. and Stevia rebaudiana (Bertoni) Bertoni, which inhibited C. gloeosporioides spores germination. After dilution of some active extracts with aqueous 1 % Tween 80 solution in a 1:2 or 1:3 ratio (extract:aqueous solution), their antifungal activity vanished. Some of the active extracts were also submitted to freeze drying and none of them presented any alteration in their antifungal activity. Concluding, several plants presented potential to be used in the search for new bioactive substances to control C. gloeosporioides, especially O. manjorona L., which inhibited 96 % of the fungus spores germination.

scholarly journals In vitro and in vivo toxicity of fungicides and biofungicides for the control of verticillium and fusarium wilt of pepper

2021 ◽  
Vol 36 (1) ◽  
pp. 23-34
Author(s):  
Milica Mihajlovic ◽  
Emil Rekanovic ◽  
Jovana Hrustic ◽  
Mila Grahovac ◽  
Brankica Tanovic
Keyword(s):  
Antifungal Activity ◽  
Verticillium Dahliae ◽  
Antagonistic Activity ◽  
In Vitro Assays ◽  
Thiophanate Methyl ◽  
Significant Difference ◽  
By Products ◽  
Pepper Plants

A survey of in vitro and in vivo sensitivity of Verticillium dahliae and Fusarium oxysporum to several commercial fungicides and biofungicides was undertaken. In in vitro assays, the tested isolate of V. dahliae proved to be very sensitive to difenoconazole (EC50 = 0.02 mg/l). However, under greenhouse conditions, the highest efficacy in V. dahliae control on inoculated pepper plants was recorded for a product based on thiophanate-methyl (83.10% compared to control). Among the tested fungicides, the lowest efficacy was recorded for a product based on azoxystrobin (23.10 %) with no significant difference compared to control (p > 0.05). In in vitro assays, the tested F. oxysporum isolate was the most sensitive to prochloraz (EC50 = 0.07 mg/l) and the least sensitive to fluopyram (EC50 = 1075.01 mg/l). In in vivo assay, the highest efficacy was achieved by products based on captan (95.60%), and the lowest by a product based on thiophanate-methyl (54.40%). Antagonistic activity of the bacterium B. subtilis under laboratory conditions was not satisfying. Also, the antifungal activity and spectrum of a tested product based on tee tree oil was not efficient in suppressing pepper wilting caused by V. dahliae and F. oxysporum.

scholarly journals Oxytocin analogues with amide groups substituted by tetrazole groups in position 4, 5 or 9.

2007 ◽  
Vol 54 (4) ◽  
pp. 805-811 ◽  
Author(s):  
Michał Manturewicz ◽  
Zbigniew Grzonka ◽  
Lenka Borovicková ◽  
Jirina Slaninová
Keyword(s):  
Amino Acids ◽  
Solid Phase ◽  
Biological Activities ◽  
Synthesis Method ◽  
Oxytocin Receptor ◽  
Amide Group ◽  
Biologically Active ◽  
Side Chain ◽  
Electronic Effects

Eleven oxytocin analogues substituted in position 4, 5 or 9 by tetrazole analogues of amino acids were prepared using solid-phase peptide synthesis method and tested for rat uterotonic in vitro and pressor activities, as well as for their affinity to human oxytocin receptor. The tetrazolic group has been used as a bioisosteric substitution of carboxylic, ester or amide groups in structure-activity relationship studies of biologically active compounds. Replacement of the amide groups of Gln(4) and Asn(5) in oxytocin by tetrazole analogues of aspartic, glutamic and alpha-aminoadipic acids containing the tetrazole moiety in the side chains leads to analogues with decreased biological activities. Oxytocin analogues in which the glycine amide residue in position 9 was substituted by tetrazole analogues of glycine had diminished activities as well. The analysis of differences in rat uterotonic activity and in the affinity to human oxytocin receptors of analogues containing either an acidic 5-substituted tetrazolic group or a neutral 1,5- or 2,5-tetrazole nucleus makes it possible to draw some new conclusions concerning the role of the amide group of amino acids in positions 4, 5 and 9 of oxytocin for its activity. The data suggest that the interaction of the side chain of Gln(4) with the oxytocin receptor is influenced mainly by electronic effects and the hydrogen bonding capacity of the amide group. Steric effects of the side chain are minor. Substitution of Asn(5) by its tetrazole derivative gave an analogue of very low activity. The result suggests that in the interaction between the amide group of Asn(5) and the binding sites of oxytocic receptor hydrogen bonds are of less importance than the spatial requirements for this group.

scholarly journals Mutational analysis of yeast profilin.

1993 ◽  
Vol 13 (12) ◽  
pp. 7864-7873 ◽  
Author(s):  
B K Haarer ◽  
A S Petzold ◽  
S S Brown
Keyword(s):  
Amino Acids ◽  
Adenylate Cyclase ◽  
Mutational Analysis ◽  
Actin Binding ◽  
In Vitro Assays ◽  
C Terminus ◽  
Physiological Relevance ◽  
In Vivo Effects

We have mutated two regions within the yeast profilin gene in an effort to functionally dissect the roles of actin and phosphatidylinositol 4,5-bisphosphate (PIP2) binding in profilin function. A series of truncations was carried out at the C terminus of profilin, a region that has been implicated in actin binding. Removal of the last three amino acids nearly eliminated the ability of profilin to bind polyproline in vitro but had no dramatic in vivo effects. Thus, the extreme C terminus is implicated in polyproline binding, but the physiological relevance of this interaction is called into question. More extensive truncation, of up to eight amino acids, had in vivo effects of increasing severity and resulted in changes in conformation and expression level of the mutant profilins. However, the ability of these mutants to bind actin in vitro was not eliminated, suggesting that this region cannot be solely responsible for actin binding. We also mutagenized a region of profilin that we hypothesized might be involved in PIP2 binding. Alteration of basic amino acids in this region produced mutant profilins that functioned well in vivo. Many of these mutants, however, were unable to suppress the loss of adenylate cyclase-associated protein (Cap/Srv2p [A. Vojtek, B. Haarer, J. Field, J. Gerst, T. D. Pollard, S. S. Brown, and M. Wigler, Cell 66:497-505, 1991]), indicating that a defect could be demonstrated in vivo. In vitro assays demonstrated that the inability to suppress loss of Cap/Srv2p correlated with a defect in the interaction with actin, independently of whether PIP2 binding was reduced. Since our earlier studies of Acanthamoeba profilins suggested the importance of PIP2 binding for suppression, we conclude that both activities are implicated and that an interplay between PIP2 binding and actin binding may be important for profilin function.

scholarly journals CD45 and the immune response.

1993 ◽  
Vol 4 (4) ◽  
pp. 976-985 ◽  
Author(s):  
J A Donovan ◽  
G A Koretzky
Keyword(s):  
Signal Transduction ◽  
Amino Acids ◽  
Immune System ◽  
Single Gene ◽  
Tyrosine Phosphatase ◽  
Consensus Sequence ◽  
Extracellular Domain ◽  
In Vitro Assays

CD45 is a major transmembrane glycoprotein expressed on all nucleated hematopoietic cells. Eight isoforms of CD45 are distributed through the immune system according to cell type and degree of cellular differentiation. Heterogeneity among the isoforms is found entirely in the extracellular domain, arising from the differential splicing of up to four exons of a single gene. The control of isoform expression suggests that the extracellular domain may participate in protein-protein interactions with isoform-specific ligands. The intracellular domain of CD45 is large (approximately 700 amino acids), identical for all isoforms, and highly conserved across species. Two nonidentical intracellular sequences of about 240 amino acids that are homologous with a tyrosine phosphatase consensus sequence have been identified. Studies with purified CD45 have shown that all isoforms possess enzymatic activity in in vitro assays. In several T and B cell lines and in natural killer cells, it appears that CD45 is required for optimal signal transduction after stimulation through a number of surface receptors. Although an in vivo substrate has not been identified conclusively, one model suggests that CD45 functions to dephosphorylate a negative-regulatory tyrosine residue on one or more protein tyrosine kinases involved in receptor-mediated second messenger formation. In T cells, the src family kinases, lck and fyn, are candidates for this regulated kinase. In this review, some of the structural and functional aspects of CD45 and its role in signal transduction in the immune system are discussed.

scholarly journals Structural and functional characterization of NEMO cleavage by SARS-CoV-2 3CLpro

2021 ◽  
Author(s):  
Mikhail Ali Hameedi ◽  
Erica Teixeira Prates ◽  
Michael R Garvin ◽  
Irimpan Mathews ◽  
B Kirtley Amos ◽  
...  
Keyword(s):  
Hydrogen Bonds ◽  
Functional Characterization ◽  
In Vitro Assays ◽  
Side Chain ◽  
Signaling Proteins ◽  
Binding Residues ◽  
Human Immune ◽  
Viral Polyprotein

In addition to its essential role in viral polyprotein processing, the SARS-CoV-2 3C-like (3CLpro) protease can cleave human immune signaling proteins, like NF-κB Essential Modulator (NEMO) and deregulate the host immune response. Here, in vitro assays show that SARS-CoV-2 3CLpro cleaves NEMO with fine-tuned efficiency. Analysis of the 2.14 Å resolution crystal structure of 3CLpro C145S bound to NEMO226-235 reveals subsites that tolerate a range of viral and host substrates through main chain hydrogen bonds while also enforcing specificity using side chain hydrogen bonds and hydrophobic contacts. Machine learning- and physics-based computational methods predict that variation in key binding residues of 3CLpro-NEMO helps explain the high fitness of SARS-CoV-2 in humans. We posit that cleavage of NEMO is an important piece of information to be accounted for in the pathology of COVID-19.

scholarly journals Synthesis and Inhibition Activity Study of Triazinyl-Substituted Amino(alkyl)-benzenesulfonamide Conjugates with Polar and Hydrophobic Amino Acids as Inhibitors of Human Carbonic Anhydrases I, II, IV, IX, and XII

2021 ◽  
Vol 22 (20) ◽  
pp. 11283
Author(s):  
Mária Bodnár Mikulová ◽  
Dáša Kružlicová ◽  
Daniel Pecher ◽  
Andrea Petreni ◽  
Claudiu T. Supuran ◽  
...  
Keyword(s):  
Amino Acids ◽  
Side Chain ◽  
Carbonic Anhydrases ◽  
Inhibition Activity ◽  
Short Reaction Time ◽  
High Affinity ◽  
Hydrophobic Amino Acids ◽  
In Vitro Inhibition ◽  
Inhibition Studies

Primary sulfonamide derivatives with various heterocycles represent the most widespread group of potential human carbonic anhydrase (hCA) inhibitors with high affinity and selectivity towards specific isozymes from the hCA family. In this work, new 4-aminomethyl- and aminoethyl-benzenesulfonamide derivatives with 1,3,5-triazine disubstituted with a pair of identical amino acids, possessing a polar (Ser, Thr, Asn, Gln) and non-polar (Ala, Tyr, Trp) side chain, have been synthesized. The optimized synthetic, purification, and isolation procedures provided several pronounced benefits such as a short reaction time (in sodium bicarbonate aqueous medium), satisfactory yields for the majority of new products (20.6–91.8%, average 60.4%), an effective, well defined semi-preparative RP-C18 liquid chromatography (LC) isolation of desired products with a high purity (>97%), as well as preservation of green chemistry principles. These newly synthesized conjugates, plus their 4-aminobenzenesulfonamide analogues prepared previously, have been investigated in in vitro inhibition studies towards hCA I, II, IV and tumor-associated isozymes IX and XII. The experimental results revealed the strongest inhibition of hCA XII with low nanomolar inhibitory constants (Kis) for the derivatives with amino acids possessing non-polar side chains (7.5–9.6 nM). Various derivatives were also promising for some other isozymes.

scholarly journals Phytochemical composition, antifungal activity, in vitro and in vivo toxicity of Syzygium cumini (L.) Skeels leaves extract

2021 ◽  
Vol 20 (5) ◽  
pp. 536-555
Author(s):  
Ernani Canuto Figueiredo Junior ◽  
◽  
Yuri Wanderley Cavalcanti ◽  
Andressa Brito Lira ◽  
Hilzeth de Luna Freire Pessoa ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Antifungal Activity ◽  
Galleria Mellonella ◽  
In Vitro Assays ◽  
Syzygium Cumini ◽  
Candida Spp ◽  
In Vivo Toxicity ◽  
Phytochemical Composition

This study determined phytochemical composition, antifungal activity and toxicity in vitro and in vivo of Syzygium cumini leaves extract (Sc). Thus, was characterized by gas chromatography coupled to mass spectrometry and submitted to determination of Minimum Inhibitory (MIC) and Fungicidal concentrations (MFC) on reference and clinical strains of Candida spp. and by growth kinetics assays. Toxicity was verified using in vitro assays of hemolysis, osmotic fragility, oxidant and antioxidant activity in human erythrocytes and by in vivo acute systemic toxicity in Galleria mellonella larvae. Fourteen different compounds were identified in Sc, which showed antifungal activity (MIC between 31.25-125 μg/mL) with fungistatic effect on Candida. At antifungal concentrations, it demonstrated low cytotoxicity, antioxidant activity and neglible in vivo toxicity. Thus, Sc demonstrated a promising antifungal potential, with low toxicity, indicating that this extract can be a safe and effective alternative antifungal agent.

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