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Antibiotic resistance is an increasing global problem and therapeutic alternatives to traditional antibiotics are needed. Antimicrobial and host defense peptides represent an attractive source for new therapeutic strategies, given their wide range of activities including antimicrobial, antitumoral and immunomodulatory. Insects produce several families of these peptides, including cecropins. Herein, we characterized the sequence, structure, and biological activity of three cecropins called satanin 1, 2, and curvicin, found in the transcriptome of two dung beetle species Dichotomius satanas and Onthophagus curvicornis. Sequence and circular dichroism analyses show that they have typical features of the cecropin family: short length (38–39 amino acids), positive charge, and amphipathic α-helical structure. They are active mainly against Gram-negative bacteria (3.12–12.5 μg/mL), with low toxicity on eukaryotic cells resulting in high therapeutic indexes (TI > 30). Peptides also showed effects on TNFα production in LPS-stimulated PBMCs. The biological activity of Satanin 1, 2 and Curvicin makes them interesting leads for antimicrobial strategies.
Diana Carolina Henao Arias; Lily Johana Toro; Germán Alberto Téllez Ramirez; Juan Felipe Osorio-Méndez; Adrián Rodríguez-Carlos; Javier Valle; Sara Paulina Marín-Luevano; Bruno Rivas-Santiago; David Andreu; Jhon Carlos Castaño Osorio. Novel antimicrobial cecropins derived from O. curvicornis and D. satanas dung beetles. Peptides 2021, 145, 170626 .
AMA StyleDiana Carolina Henao Arias, Lily Johana Toro, Germán Alberto Téllez Ramirez, Juan Felipe Osorio-Méndez, Adrián Rodríguez-Carlos, Javier Valle, Sara Paulina Marín-Luevano, Bruno Rivas-Santiago, David Andreu, Jhon Carlos Castaño Osorio. Novel antimicrobial cecropins derived from O. curvicornis and D. satanas dung beetles. Peptides. 2021; 145 ():170626.
Chicago/Turabian StyleDiana Carolina Henao Arias; Lily Johana Toro; Germán Alberto Téllez Ramirez; Juan Felipe Osorio-Méndez; Adrián Rodríguez-Carlos; Javier Valle; Sara Paulina Marín-Luevano; Bruno Rivas-Santiago; David Andreu; Jhon Carlos Castaño Osorio. 2021. "Novel antimicrobial cecropins derived from O. curvicornis and D. satanas dung beetles." Peptides 145, no. : 170626.
An approach based on a dendrimer display of B- and T-cell epitopes relevant for antibody induction has been shown to be effective as a foot-and-mouth disease (FMD) vaccine. B2T dendrimers combining two copies of the major FMD virus (FMDV) type O B-cell epitope (capsid proteinVP1 (140–158)) covalently linked to a heterotypic T-cell epitope from non-structural protein 3A (21–35), henceforth B2T-3A, has previously been shown to elicit high neutralizing antibody (nAb) titers and IFN-γ-producing cells in both mice and pigs. Here, we provide evidence that the B- and T-cell epitopes need to be tethered to a single molecular platform for successful T-cell help, leading to efficient nAb induction in mice. In addition, mice immunized with a non-covalent mixture of B2T-3A dendrimers containing the B-cell epitopes of FMDV types O and C induced similarly high nAb levels against both serotypes, opening the way for a multivalent vaccine platform against a variety of serologically different FMDVs. These findings are relevant for the design of vaccine strategies based on B- and T-cell epitope combinations.
Rodrigo Cañas-Arranz; Patricia de León; Sira Defaus; Elisa Torres; Mar Forner; María Bustos; David Andreu; Esther Blanco; Francisco Sobrino. Immunogenicity of Foot-and-Mouth Disease Virus Dendrimer Peptides: Need for a T-Cell Epitope and Ability to Elicit Heterotypic Responses. Molecules 2021, 26, 4714 .
AMA StyleRodrigo Cañas-Arranz, Patricia de León, Sira Defaus, Elisa Torres, Mar Forner, María Bustos, David Andreu, Esther Blanco, Francisco Sobrino. Immunogenicity of Foot-and-Mouth Disease Virus Dendrimer Peptides: Need for a T-Cell Epitope and Ability to Elicit Heterotypic Responses. Molecules. 2021; 26 (16):4714.
Chicago/Turabian StyleRodrigo Cañas-Arranz; Patricia de León; Sira Defaus; Elisa Torres; Mar Forner; María Bustos; David Andreu; Esther Blanco; Francisco Sobrino. 2021. "Immunogenicity of Foot-and-Mouth Disease Virus Dendrimer Peptides: Need for a T-Cell Epitope and Ability to Elicit Heterotypic Responses." Molecules 26, no. 16: 4714.
Multidrug resistance against conventional antibiotics poses an important threat to human health. In this context, antimicrobial peptides (AMPs) have been extensively studied for their antibacterial activity and promising results have been shown so far. However, AMPs tend to be rather vulnerable to protease degradation, which offsets their therapeutic appeal. Here, we demonstrate how replacing functional residues in the antimicrobial region of human RNase 3—also named eosinophil cationic protein—by non-natural amino acids increases stability in human serum. These changes were also shown to reduce the hemolytic effect of the peptides in general terms, whereas the antimicrobial activity was reasonably preserved. Digestion profiles enabled us to design new peptides with superior stability and lower toxicity that could become relevant candidates to reach clinical stages.
Daniel Sandín; Javier Valle; Belén Chaves-Arquero; Guillem Prats-Ejarque; María Nieves Larrosa; Juan José González-López; María Ángeles Jiménez; Ester Boix; David Andreu; Marc Torrent. Rationally Modified Antimicrobial Peptides from the N-Terminal Domain of Human RNase 3 Show Exceptional Serum Stability. Journal of Medicinal Chemistry 2021, 1 .
AMA StyleDaniel Sandín, Javier Valle, Belén Chaves-Arquero, Guillem Prats-Ejarque, María Nieves Larrosa, Juan José González-López, María Ángeles Jiménez, Ester Boix, David Andreu, Marc Torrent. Rationally Modified Antimicrobial Peptides from the N-Terminal Domain of Human RNase 3 Show Exceptional Serum Stability. Journal of Medicinal Chemistry. 2021; ():1.
Chicago/Turabian StyleDaniel Sandín; Javier Valle; Belén Chaves-Arquero; Guillem Prats-Ejarque; María Nieves Larrosa; Juan José González-López; María Ángeles Jiménez; Ester Boix; David Andreu; Marc Torrent. 2021. "Rationally Modified Antimicrobial Peptides from the N-Terminal Domain of Human RNase 3 Show Exceptional Serum Stability." Journal of Medicinal Chemistry , no. : 1.
Passing through the blood-brain barrier (BBB) to treat neurological conditions is one of the main hurdles in modern medicine. Many drugs with promising in vitro profiles become ineffective in vivo due to BBB restrictive permeability. In particular, this includes drugs such as antiviral porphyrins, with the ability to fight brain-resident viruses causing diseases such as HIV-associated neurocognitive disorders (HAND). In the last two decades, BBB shuttles, particularly peptide-based ones, have shown promise in carrying various payloads across the BBB. Thus, peptide–drug conjugates (PDCs) formed by covalent attachment of a BBB peptide shuttle and an antiviral drug may become key therapeutic tools in treating neurological disorders of viral origin. In this study, we have used various approaches (guanidinium, phosphonium, and carbodiimide-based couplings) for on-resin synthesis of new peptide–porphyrin conjugates (PPCs) with BBB-crossing and potential antiviral activity. After careful fine-tuning of the synthetic chemistry, DIC/oxyma has emerged as a preferred method, by which 14 different PPCs have been made and satisfactorily characterized. The PPCs are prepared by coupling a porphyrin carboxyl group to an amino group (either N-terminal or a Lys side chain) of the peptide shuttle and show effective in vitro BBB translocation ability, low cytotoxicity toward mouse brain endothelial cells, and low hemolytic activity. Three of the PPCs, MP-P5, P4-MP, and P4-L-MP, effectively inhibiting HIV infectivity in vitro, stand out as most promising. Their efficacy against other brain-targeting viruses (Dengue, Zika, and SARS-CoV-2) is currently under evaluation, with preliminary results confirming that PPCs are a promising strategy to treat viral brain infections.
Diogo A. Mendonça; Mariët Bakker; Christine Cruz-Oliveira; Vera Neves; Maria Angeles Jiménez; Sira Defaus; Marco Cavaco; Ana Salomé Veiga; Iris Cadima-Couto; Miguel A. R. B. Castanho; David Andreu; Toni Todorovski. Penetrating the Blood-Brain Barrier with New Peptide–Porphyrin Conjugates Having anti-HIV Activity. Bioconjugate Chemistry 2021, 32, 1067 -1077.
AMA StyleDiogo A. Mendonça, Mariët Bakker, Christine Cruz-Oliveira, Vera Neves, Maria Angeles Jiménez, Sira Defaus, Marco Cavaco, Ana Salomé Veiga, Iris Cadima-Couto, Miguel A. R. B. Castanho, David Andreu, Toni Todorovski. Penetrating the Blood-Brain Barrier with New Peptide–Porphyrin Conjugates Having anti-HIV Activity. Bioconjugate Chemistry. 2021; 32 (6):1067-1077.
Chicago/Turabian StyleDiogo A. Mendonça; Mariët Bakker; Christine Cruz-Oliveira; Vera Neves; Maria Angeles Jiménez; Sira Defaus; Marco Cavaco; Ana Salomé Veiga; Iris Cadima-Couto; Miguel A. R. B. Castanho; David Andreu; Toni Todorovski. 2021. "Penetrating the Blood-Brain Barrier with New Peptide–Porphyrin Conjugates Having anti-HIV Activity." Bioconjugate Chemistry 32, no. 6: 1067-1077.
Vaccines are considered one of the greatest global health achievements, improving the welfare of society by saving lives and substantially reducing the burden of infectious diseases. However, few vaccines are fully effective, for reasons ranging from intrinsic limitations to more contingent shortcomings related, e.g., to cold chain transport, handling and storage. In this context, subunit vaccines where the essential antigenic traits (but not the entire pathogen) are presented in rationally designed fashion have emerged as an attractive alternative to conventional ones. In particular, this includes the option of fully synthetic peptide vaccines able to mimic well-defined B- and T-cell epitopes from the infectious agent and to induce protection against it. Although, in general, linear peptides have been associated to low immunogenicity and partial protection, there are several strategies to address such issues. In this review, we report the progress towards the development of peptide-based vaccines against foot-and-mouth disease (FMD) a highly transmissible, economically devastating animal disease. Starting from preliminary experiments using single linear B-cell epitopes, recent research has led to more complex and successful second-generation vaccines featuring peptide dendrimers containing multiple copies of B- and T-cell epitopes against FMD virus or classical swine fever virus (CSFV). The usefulness of this strategy to prevent other animal and human diseases is discussed.
Mar Forner; Rodrigo Cañas-Arranz; Sira Defaus; Patricia de León; Miguel Rodríguez-Pulido; Llilianne Ganges; Esther Blanco; Francisco Sobrino; David Andreu. Peptide-Based Vaccines: Foot-and-Mouth Disease Virus, a Paradigm in Animal Health. Vaccines 2021, 9, 477 .
AMA StyleMar Forner, Rodrigo Cañas-Arranz, Sira Defaus, Patricia de León, Miguel Rodríguez-Pulido, Llilianne Ganges, Esther Blanco, Francisco Sobrino, David Andreu. Peptide-Based Vaccines: Foot-and-Mouth Disease Virus, a Paradigm in Animal Health. Vaccines. 2021; 9 (5):477.
Chicago/Turabian StyleMar Forner; Rodrigo Cañas-Arranz; Sira Defaus; Patricia de León; Miguel Rodríguez-Pulido; Llilianne Ganges; Esther Blanco; Francisco Sobrino; David Andreu. 2021. "Peptide-Based Vaccines: Foot-and-Mouth Disease Virus, a Paradigm in Animal Health." Vaccines 9, no. 5: 477.
The activation of cannabinoid CB1 receptors (CB1R) by Δ9-tetrahydrocannabinol (THC), the main component of Cannabis sativa, induces analgesia. CB1R activation, however, also causes cognitive impairment via the serotonin 5HT2A receptor (5HT2AR), a component of a CB1R–5HT2AR heteromer, posing a serious drawback for cannabinoid therapeutic use. We have shown that peptides reproducing CB1R transmembrane (TM) helices 5 and 6, fused to a cell-penetrating sequence (CPP), can alter the structure of the CB1R–5HT2AR heteromer and avert THC cognitive impairment while preserving analgesia. Here, we report the optimization of these prototypes into drug-like leads by (i) shortening the TM5, TM6, and CPP sequences, without losing the ability to disturb the CB1R–5HT2AR heteromer, and (ii) extensive sequence remodeling to achieve protease resistance and blood–brain barrier penetration. Our efforts have culminated in the identification of an ideal candidate for cannabis-based pain management, an orally active 16-residue peptide preserving THC-induced analgesia.
Maria Gallo; Estefanía Moreno; Sira Defaus; Antonio Ortega-Alvaro; Angel Gonzalez; Patricia Robledo; Marco Cavaco; Vera Neves; Miguel A. R. B. Castanho; Vicent Casadó; Leonardo Pardo; Rafael Maldonado; David Andreu. Orally Active Peptide Vector Allows Using Cannabis to Fight Pain While Avoiding Side Effects. Journal of Medicinal Chemistry 2021, 64, 6937 -6948.
AMA StyleMaria Gallo, Estefanía Moreno, Sira Defaus, Antonio Ortega-Alvaro, Angel Gonzalez, Patricia Robledo, Marco Cavaco, Vera Neves, Miguel A. R. B. Castanho, Vicent Casadó, Leonardo Pardo, Rafael Maldonado, David Andreu. Orally Active Peptide Vector Allows Using Cannabis to Fight Pain While Avoiding Side Effects. Journal of Medicinal Chemistry. 2021; 64 (10):6937-6948.
Chicago/Turabian StyleMaria Gallo; Estefanía Moreno; Sira Defaus; Antonio Ortega-Alvaro; Angel Gonzalez; Patricia Robledo; Marco Cavaco; Vera Neves; Miguel A. R. B. Castanho; Vicent Casadó; Leonardo Pardo; Rafael Maldonado; David Andreu. 2021. "Orally Active Peptide Vector Allows Using Cannabis to Fight Pain While Avoiding Side Effects." Journal of Medicinal Chemistry 64, no. 10: 6937-6948.
Proteolytic instability is a critical limitation for peptide‐based products. Although significant efforts are devoted to stabilize sequences against proteases/peptidases in plasma/serum, such approaches tend to be rather empirical, unspecific, time‐consuming, and frequently not cost‐effective. A more rational and potentially rewarding alternative is to identify the chemical grounds of susceptibility to enzymatic degradation of peptides so that proteolytic resistance can be tuned by manipulation of key chemical properties. In this regard, we conducted a meta‐analysis of literature published over the last decade reporting experimental data on the lifetimes of peptides exposed to proteolytic conditions. Our initial database contained 579 entries and was curated with regard to amino acid sequence, chemical modification, terminal half‐life (t1/2) or other stability readouts, type of stability assay, and biological application of the study. Although the majority of entries in the database corresponded to (slightly or substantially) modified peptides, we chose to focus on unmodified ones, as we aimed to decipher intrinsic characteristics of peptide proteolytic susceptibility. Specifically, we developed a multivariable regression model to unravel those peptide properties with most impact on proteolytic stability and thus potential t1/2 predicting ability. Model validation was done by two different approaches. First, a library of peptides spanning a large interval of properties that modulate stability was synthesized and their t1/2 in human serum were experimentally determined. Second, the t1/2 of 21 selected peptides approved for clinical use or in clinical trials were recorded and matched with the model‐estimated values. With both approaches, good correlation between experimental and predicted t1/2 data was observed.
Marco Cavaco; Javier Valle; Isabel Flores; David Andreu; Miguel A. R. B. Castanho. Estimating peptide half‐life in serum from tunable, sequence‐related physicochemical properties. Clinical and Translational Science 2021, 14, 1349 -1358.
AMA StyleMarco Cavaco, Javier Valle, Isabel Flores, David Andreu, Miguel A. R. B. Castanho. Estimating peptide half‐life in serum from tunable, sequence‐related physicochemical properties. Clinical and Translational Science. 2021; 14 (4):1349-1358.
Chicago/Turabian StyleMarco Cavaco; Javier Valle; Isabel Flores; David Andreu; Miguel A. R. B. Castanho. 2021. "Estimating peptide half‐life in serum from tunable, sequence‐related physicochemical properties." Clinical and Translational Science 14, no. 4: 1349-1358.
The tumour necrosis factor-like weak inducer of apoptosis (TWEAK) is a member of the tumour necrosis factor ligand family and has been shown to be overexpressed in tumoral cells together with the fibroblast growth factor–inducible 14 (Fn14) receptor. TWEAK-Fn14 interaction triggers a set of intracellular pathways responsible for tumour cell invasion and migration, as well as proliferation and angiogenesis. Hence, modulation of the TWEAK-Fn14 interaction is an important therapeutic goal. The targeting of protein-protein interactions by external agents, e.g., drugs, remains a substantial challenge. Given their intrinsic features, as well as recent advances that improve their pharmacological profiles, peptides have arisen as promising agents in this regard. Here, we report, by in silico structural design validated by cell-based and in vitro assays, the discovery of four peptides able to target TWEAK. Our results show that, when added to TWEAK-dependent cellular cultures, peptides cause a down-regulation of genes that are part of TWEAK-Fn14 signalling pathway. The direct, physical interaction between the peptides and TWEAK was further elucidated in an in vitro assay which confirmed that the bioactivity shown in cell-based assays was due to the targeting of TWEAK. The results presented here are framed within early pre-clinical drug development and therefore these peptide hits represent a starting point for the development of novel therapeutic agents. Our approach exemplifies the powerful combination of in silico and experimental efforts to quickly identify peptides with desirable traits.
Miriam Badia-Villanueva; Sira Defaus; Ruben Foj; David Andreu; Baldo Oliva; Angels Sierra; Narcis Fernandez-Fuentes. Evaluation of Computationally Designed Peptides Against TWEAK, a Cytokine of the Tumour Necrosis Factor Ligand Family. International Journal of Molecular Sciences 2021, 22, 1066 .
AMA StyleMiriam Badia-Villanueva, Sira Defaus, Ruben Foj, David Andreu, Baldo Oliva, Angels Sierra, Narcis Fernandez-Fuentes. Evaluation of Computationally Designed Peptides Against TWEAK, a Cytokine of the Tumour Necrosis Factor Ligand Family. International Journal of Molecular Sciences. 2021; 22 (3):1066.
Chicago/Turabian StyleMiriam Badia-Villanueva; Sira Defaus; Ruben Foj; David Andreu; Baldo Oliva; Angels Sierra; Narcis Fernandez-Fuentes. 2021. "Evaluation of Computationally Designed Peptides Against TWEAK, a Cytokine of the Tumour Necrosis Factor Ligand Family." International Journal of Molecular Sciences 22, no. 3: 1066.
Conjugation of TP10, a cell-penetrating peptide with intrinsic antimalarial activity, to the well-known antimalarial drugs chloroquine and primaquine has been previously shown to enhance the peptide’s action against, respectively, blood- and liver-stage malaria parasites. Yet, this was achieved at the cost of a significant increase in haemolytic activity, as fluorescence microscopy and flow cytometry studies showed the conjugates to be more haemolytic for non-infected than for Plasmodium-infected red blood cells. To gain further insight into how these conjugates distinctively bind, and likely disrupt, membranes of both Plasmodium-infected and non-infected erythrocytes, we used dynamic light scattering and surface plasmon resonance to study the interactions of two representative conjugates and their parent compounds with lipid model membranes. Results obtained are herein reported and confirm that a strong membrane-disruptive character underlies the haemolytic properties of these conjugates, thus hampering their ability to exert selective antimalarial action.
Luísa Aguiar; Marina Pinheiro; Ana Rute Neves; Nuno Vale; Sira Defaus; David Andreu; Salette Reis; Paula Gomes. Insights into the Membranolytic Activity of Antimalarial Drug-Cell Penetrating Peptide Conjugates. Membranes 2020, 11, 4 .
AMA StyleLuísa Aguiar, Marina Pinheiro, Ana Rute Neves, Nuno Vale, Sira Defaus, David Andreu, Salette Reis, Paula Gomes. Insights into the Membranolytic Activity of Antimalarial Drug-Cell Penetrating Peptide Conjugates. Membranes. 2020; 11 (1):4.
Chicago/Turabian StyleLuísa Aguiar; Marina Pinheiro; Ana Rute Neves; Nuno Vale; Sira Defaus; David Andreu; Salette Reis; Paula Gomes. 2020. "Insights into the Membranolytic Activity of Antimalarial Drug-Cell Penetrating Peptide Conjugates." Membranes 11, no. 1: 4.
There is an urgent need for the development of new anti-HIV drugs that can complement existing medicines to be used against resistant strains. Here, we report the anti-HIV-1 peptide pepRF1, a human serum-resistant peptide derived from the Dengue virus capsid protein. In vitro, pepRF1 shows a 50% inhibitory concentration of 1.5 nM with a potential therapeutic window higher than 53 000. This peptide is specific for CXCR4-tropic strains, preventing viral entry into target cells by binding to the viral coreceptor CXCR4, acting as an antagonist of this receptor. pepRF1 is more effective than T20, the only peptide-based HIV-1 entry inhibitor approved, and excels in inhibiting a HIV-1 strain resistant to T20. Potentially, pepRF1 can be used alone or in combination with other anti-HIV drugs. Furthermore, one can also envisage its use as a novel therapeutic strategy for other CXCR4-related diseases.
Iris Cadima-Couto; Alexandra Tauzin; João M. Freire; Tiago N. Figueira; Rúben D. M. Silva; Clara Pérez-Peinado; Catarina Cunha-Santos; Inês Bártolo; Nuno Taveira; Lurdes Gano; João D. G. Correia; Joao Goncalves; Fabrizio Mammano; David Andreu; Miguel A. R. B. Castanho; Ana Salomé Veiga. Anti-HIV-1 Activity of pepRF1, a Proteolysis-Resistant CXCR4 Antagonist Derived from Dengue Virus Capsid Protein. ACS Infectious Diseases 2020, 7, 6 -22.
AMA StyleIris Cadima-Couto, Alexandra Tauzin, João M. Freire, Tiago N. Figueira, Rúben D. M. Silva, Clara Pérez-Peinado, Catarina Cunha-Santos, Inês Bártolo, Nuno Taveira, Lurdes Gano, João D. G. Correia, Joao Goncalves, Fabrizio Mammano, David Andreu, Miguel A. R. B. Castanho, Ana Salomé Veiga. Anti-HIV-1 Activity of pepRF1, a Proteolysis-Resistant CXCR4 Antagonist Derived from Dengue Virus Capsid Protein. ACS Infectious Diseases. 2020; 7 (1):6-22.
Chicago/Turabian StyleIris Cadima-Couto; Alexandra Tauzin; João M. Freire; Tiago N. Figueira; Rúben D. M. Silva; Clara Pérez-Peinado; Catarina Cunha-Santos; Inês Bártolo; Nuno Taveira; Lurdes Gano; João D. G. Correia; Joao Goncalves; Fabrizio Mammano; David Andreu; Miguel A. R. B. Castanho; Ana Salomé Veiga. 2020. "Anti-HIV-1 Activity of pepRF1, a Proteolysis-Resistant CXCR4 Antagonist Derived from Dengue Virus Capsid Protein." ACS Infectious Diseases 7, no. 1: 6-22.
Proteolytic stability assessment is increasingly viewed as a fundamental component of peptide characterization, arguably of comparable importance as efficacy and toxicity data. A literature survey over the last decade reveals steady growth in the stability information available. However, it also uncovers two significant problems that hinder proper data comparison: 1) the use of different stability assays, and 2) the differences in how stability information is reported. In this Viewpoint, we present results from a database meta‐analysis as well as concerns about the stability assessments published so far. We also suggest guidelines for a proper discussion between experts in the field on how to improve data readability so that peptide stability, an often‐missing parameter in older literature, is adequately reported to take maximum advantage of it.
Marco Cavaco; David Andreu; Miguel A. R. B. Castanho. The Challenge of Peptide Proteolytic Stability Studies: Scarce Data, Difficult Readability, and the Need for Harmonization. Angewandte Chemie International Edition 2020, 60, 1686 -1688.
AMA StyleMarco Cavaco, David Andreu, Miguel A. R. B. Castanho. The Challenge of Peptide Proteolytic Stability Studies: Scarce Data, Difficult Readability, and the Need for Harmonization. Angewandte Chemie International Edition. 2020; 60 (4):1686-1688.
Chicago/Turabian StyleMarco Cavaco; David Andreu; Miguel A. R. B. Castanho. 2020. "The Challenge of Peptide Proteolytic Stability Studies: Scarce Data, Difficult Readability, and the Need for Harmonization." Angewandte Chemie International Edition 60, no. 4: 1686-1688.
Proteolytic stability assessment is increasingly viewed as a fundamental component of peptide characterization, arguably of comparable importance as efficacy and toxicity data. A literature survey over the last decade reveals steady growth in the stability information available. However, it also uncovers two significant problems that hinder proper data comparison: 1) the use of different stability assays, and 2) the differences in how stability information is reported. In this Viewpoint, we present results from a database meta‐analysis as well as concerns about the stability assessments published so far. We also suggest guidelines for a proper discussion between experts in the field on how to improve data readability so that peptide stability, an often‐missing parameter in older literature, is adequately reported to take maximum advantage of it.
Marco Cavaco; David Andreu; Miguel A. R. B. Castanho. The Challenge of Peptide Proteolytic Stability Studies: Scarce Data, Difficult Readability, and the Need for Harmonization. Angewandte Chemie 2020, 133, 1710 -1712.
AMA StyleMarco Cavaco, David Andreu, Miguel A. R. B. Castanho. The Challenge of Peptide Proteolytic Stability Studies: Scarce Data, Difficult Readability, and the Need for Harmonization. Angewandte Chemie. 2020; 133 (4):1710-1712.
Chicago/Turabian StyleMarco Cavaco; David Andreu; Miguel A. R. B. Castanho. 2020. "The Challenge of Peptide Proteolytic Stability Studies: Scarce Data, Difficult Readability, and the Need for Harmonization." Angewandte Chemie 133, no. 4: 1710-1712.
Ctn[15–34], the C-terminal fragment of crotalicidin, an antimicrobial peptide from the South American rattlesnake Crotalus durissus terrificus venom, displays remarkable anti-infective and anti-proliferative activities. Herein, its activity on Candida albicans biofilms and its interaction with the cytoplasmic membrane of the fungal cell and with a biomembrane model in vitro was investigated. A standard C. albicans strain and a fluconazole-resistant clinical isolate were exposed to the peptide at its minimum inhibitory concentration (MIC) (10 µM) and up to 100 × MIC to inhibit biofilm formation and its eradication. A viability test using XTT and fluorescent dyes, confocal laser scanning microscopy, and atomic force microscopy (AFM) were used to observe the antibiofilm effect. To evaluate the importance of membrane composition on Ctn[15–34] activity, C. albicans protoplasts were also tested. Fluorescence assays using di-8-ANEPPS, dynamic light scattering, and zeta potential measurements using liposomes, protoplasts, and C. albicans cells indicated a direct mechanism of action that was dependent on membrane interaction and disruption. Overall, Ctn[15–34] showed to be an effective antifungal peptide, displaying antibiofilm activity and, importantly, interacting with and disrupting fungal plasma membrane.
Francisca Aguiar; Nuno Santos; Carolina De Paula Cavalcante; David Andreu; Gandhi Baptista; Sónia Gonçalves. Antibiofilm Activity on Candida albicans and Mechanism of Action on Biomembrane Models of the Antimicrobial Peptide Ctn[15–34]. International Journal of Molecular Sciences 2020, 21, 8339 .
AMA StyleFrancisca Aguiar, Nuno Santos, Carolina De Paula Cavalcante, David Andreu, Gandhi Baptista, Sónia Gonçalves. Antibiofilm Activity on Candida albicans and Mechanism of Action on Biomembrane Models of the Antimicrobial Peptide Ctn[15–34]. International Journal of Molecular Sciences. 2020; 21 (21):8339.
Chicago/Turabian StyleFrancisca Aguiar; Nuno Santos; Carolina De Paula Cavalcante; David Andreu; Gandhi Baptista; Sónia Gonçalves. 2020. "Antibiofilm Activity on Candida albicans and Mechanism of Action on Biomembrane Models of the Antimicrobial Peptide Ctn[15–34]." International Journal of Molecular Sciences 21, no. 21: 8339.
Crotalicidin (Ctn) and its fragment Ctn[15–34] are snake-venom-derived, cathelicidin-related peptides outstanding for their promising antimicrobial, antifungal, and antitumoral properties. In this study, we describe their membranolytic mechanisms as well as their putative interference with intracellular targets, both contributing to their antitumoral action against a pro-monocytic leukemia cell line. Initial flow cytometry assays demonstrated peptide ability to induce tumor cell membrane permeabilization and caspase-dependent apoptosis, without total activity reduction by serum proteases up to 24 h (Ctn) and 18 h (Ctn[15–34]). In addition, both Ctn and Ctn[15–34] showed preference for tumor cells rather than healthy cells, with selectivity ratios (tumoral vs healthy cells) of 17 and 7, respectively. Further microscopy and flow cytometry studies suggested their preferential accumulation in the cytoplasmic membrane and nucleus and proposed multiple predominant routes of peptide uptake, including direct entry and endocytosis. Affinity purification followed by proteomic identification experiments revealed both peptides to interact with proteins involved in DNA and protein metabolism, cell cycles, signal transduction, and/or programmed cell death, among others. These results suggest a putative role of Ctn and Ctn[15–34] to interact with key intracellular pathways, ultimately contributing to tumor cell death by necrosis/apoptosis. Altogether, this work proposes a dual mechanism underlying the antitumoral activity of Ctn and Ctn[15–34] and reinforces their potential as future therapeutic drugs.
Clara Pérez-Peinado; Javier Valle; João M. Freire; David Andreu. Tumor Cell Attack by Crotalicidin (Ctn) and Its Fragment Ctn[15–34]: Insights into Their Dual Membranolytic and Intracellular Targeting Mechanism. ACS Chemical Biology 2020, 15, 2945 -2957.
AMA StyleClara Pérez-Peinado, Javier Valle, João M. Freire, David Andreu. Tumor Cell Attack by Crotalicidin (Ctn) and Its Fragment Ctn[15–34]: Insights into Their Dual Membranolytic and Intracellular Targeting Mechanism. ACS Chemical Biology. 2020; 15 (11):2945-2957.
Chicago/Turabian StyleClara Pérez-Peinado; Javier Valle; João M. Freire; David Andreu. 2020. "Tumor Cell Attack by Crotalicidin (Ctn) and Its Fragment Ctn[15–34]: Insights into Their Dual Membranolytic and Intracellular Targeting Mechanism." ACS Chemical Biology 15, no. 11: 2945-2957.
The characterization of biologically active peptides relies heavily on the study of their efficacy, toxicity, mechanism of action, cellular uptake, or intracellular location, using both in vitro and in vivo studies. These studies frequently depend on the use of fluorescence-based techniques. Since most peptides are not intrinsically fluorescent, they are conjugated to a fluorophore. The conjugation may interfere with peptide properties, thus biasing the results. The selection of the most suitable fluorophore is highly relevant. Here, a comprehensive study with blood–brain barrier (BBB) peptide shuttles (PepH3 and PepNeg) and antimicrobial peptides (AMPs) (vCPP2319 and Ctn[15-34]), tested as anticancer peptides (ACPs), having different fluorophores, namely 5(6)-carboxyfluorescein (CF), rhodamine B (RhB), quasar 570 (Q570), or tide fluor 3 (TF3) attached is presented. The goal is the evaluation of the impact of the selected fluorophores on peptide performance, applying routinely used techniques to assess cytotoxicity/toxicity, secondary structure, BBB translocation, and cellular internalization. Our results show that some fluorophores significantly modulate peptide activity when compared with unlabeled peptides, being more noticeable in hydrophobic and charged fluorophores. This study highlights the need for a careful experimental design for fluorescently labeled molecules, such as peptides.
Marco Cavaco; Clara Pérez-Peinado; Javier Valle; Rúben D. M. Silva; João D. G. Correia; David Andreu; Miguel A. R. B. Castanho; Vera Neves. To What Extent Do Fluorophores Bias the Biological Activity of Peptides? A Practical Approach Using Membrane-Active Peptides as Models. Frontiers in Bioengineering and Biotechnology 2020, 8, 1 .
AMA StyleMarco Cavaco, Clara Pérez-Peinado, Javier Valle, Rúben D. M. Silva, João D. G. Correia, David Andreu, Miguel A. R. B. Castanho, Vera Neves. To What Extent Do Fluorophores Bias the Biological Activity of Peptides? A Practical Approach Using Membrane-Active Peptides as Models. Frontiers in Bioengineering and Biotechnology. 2020; 8 ():1.
Chicago/Turabian StyleMarco Cavaco; Clara Pérez-Peinado; Javier Valle; Rúben D. M. Silva; João D. G. Correia; David Andreu; Miguel A. R. B. Castanho; Vera Neves. 2020. "To What Extent Do Fluorophores Bias the Biological Activity of Peptides? A Practical Approach Using Membrane-Active Peptides as Models." Frontiers in Bioengineering and Biotechnology 8, no. : 1.
Dendrimer peptides are promising vaccine candidates against the foot-and-mouth disease virus (FMDV). Several B-cell epitope (B2T) dendrimers, harboring a major FMDV antigenic B-cell site in VP1 protein, are covalently linked to heterotypic T-cell epitopes from 3A and/or 3D proteins, and elicited consistent levels of neutralizing antibodies and IFN-γ-producing cells in pigs. To address the contribution of the highly polymorphic nature of the porcine MHC (SLA, swine leukocyte antigen) on the immunogenicity of B2T dendrimers, low-resolution (Lr) haplotyping was performed. We looked for possible correlations between particular Lr haplotypes with neutralizing antibody and T-cell responses induced by B2T peptides. In this study, 63 pigs immunized with B2T dendrimers and 10 non-immunized (control) animals are analyzed. The results reveal a robust significant correlation between SLA class-II Lr haplotypes and the T-cell response. Similar correlations of T-cell response with SLA class-I Lr haplotypes, and between B-cell antibody response and SLA class-I and SLA class-II Lr haplotypes, were only found when the sample was reduced to animals with Lr haplotypes represented more than once. These results support the contribution of SLA class-II restricted T-cells to the magnitude of the T-cell response and to the antibody response evoked by the B2T dendrimers, being of potential value for peptide vaccine design against FMDV.
Patricia De León; Rodrigo Cañas-Arranz; Yago Saez; Mar Forner; Sira Defaus; Dolores Cuadra; María J. Bustos; Elisa Torres; David Andreu; Esther Blanco; Francisco Sobrino; Sabine E. Hammer. Association of Porcine Swine Leukocyte Antigen (SLA) Haplotypes with B- and T-Cell Immune Response to Foot-and-Mouth Disease Virus (FMDV) Peptides. Vaccines 2020, 8, 513 .
AMA StylePatricia De León, Rodrigo Cañas-Arranz, Yago Saez, Mar Forner, Sira Defaus, Dolores Cuadra, María J. Bustos, Elisa Torres, David Andreu, Esther Blanco, Francisco Sobrino, Sabine E. Hammer. Association of Porcine Swine Leukocyte Antigen (SLA) Haplotypes with B- and T-Cell Immune Response to Foot-and-Mouth Disease Virus (FMDV) Peptides. Vaccines. 2020; 8 (3):513.
Chicago/Turabian StylePatricia De León; Rodrigo Cañas-Arranz; Yago Saez; Mar Forner; Sira Defaus; Dolores Cuadra; María J. Bustos; Elisa Torres; David Andreu; Esther Blanco; Francisco Sobrino; Sabine E. Hammer. 2020. "Association of Porcine Swine Leukocyte Antigen (SLA) Haplotypes with B- and T-Cell Immune Response to Foot-and-Mouth Disease Virus (FMDV) Peptides." Vaccines 8, no. 3: 513.
Synthetic dendrimer peptides are a promising strategy to develop new FMD vaccines. A dendrimer peptide, termed B2T-3A, which harbors two copies of the major FMDV antigenic B-cell site [VP1 (140–158)], covalently linked to a heterotypic T-cell from the non-structural protein 3A [3A (21–35)], has been shown to protect pigs against viral challenge. Interestingly, the modular design of this dendrimer peptide allows modifications aimed at improving its immunogenicity, such as the replacement of the T-cell epitope moiety. Here, we report that a dendrimer peptide, B2T-3D, harboring a T-cell epitope from FMDV 3D protein [3D (56–70)], when inoculated in pigs, elicited consistent levels of neutralizing antibodies and high frequencies of IFN-γ-producing cells upon in vitro recall with the homologous dendrimers, both responses being similar to those evoked by B2T-3A. Lymphocytes from B2T-3A-immunized pigs were in vitro-stimulated by T-3A peptide and to a lesser extent by B-peptide, while those from B2T-3D- immunized animals preferentially recognized the T-3D peptide, suggesting that this epitope is a potent inducer of IFN-γ producing-cells. These results extend the repertoire of T-cell epitopes efficiently recognized by swine lymphocytes and open the possibility of using T-3D to enhance the immunogenicity and the protection conferred by B2T-dendrimers.
Rodrigo Cañas-Arranz; Patricia De León; Mar Forner; Sira Defaus; María J. Bustos; Elisa Torres; David Andreu; Esther Blanco; Francisco Sobrino. Immunogenicity of a Dendrimer B2T Peptide Harboring a T-Cell Epitope From FMDV Non-structural Protein 3D. Frontiers in Veterinary Science 2020, 7, 498 .
AMA StyleRodrigo Cañas-Arranz, Patricia De León, Mar Forner, Sira Defaus, María J. Bustos, Elisa Torres, David Andreu, Esther Blanco, Francisco Sobrino. Immunogenicity of a Dendrimer B2T Peptide Harboring a T-Cell Epitope From FMDV Non-structural Protein 3D. Frontiers in Veterinary Science. 2020; 7 ():498.
Chicago/Turabian StyleRodrigo Cañas-Arranz; Patricia De León; Mar Forner; Sira Defaus; María J. Bustos; Elisa Torres; David Andreu; Esther Blanco; Francisco Sobrino. 2020. "Immunogenicity of a Dendrimer B2T Peptide Harboring a T-Cell Epitope From FMDV Non-structural Protein 3D." Frontiers in Veterinary Science 7, no. : 498.
Available treatments of invasive fungal infection have limitations including toxicity and the emergence of resistant strains. Therefore, there is an urgent need for alternative solutions. Due to the unique mode of action and high selectivity, plant defensins (PDs) are worthy therapeutic candidates. Chemical synthesis remains a preferred method for the production of many peptide-based therapeutics. Given the relatively long sequence of PDs, as well as their complicated posttranslational modifications, the synthetic route can be considered challenging. Here we describe a total synthesis of PvD1, the defensin from the common bean Phaseolus vulgaris. Analytical, structural and functional characterization revealed that both natural and synthetic peptides fold into a canonical CSαβ motif stabilized by conserved disulfide bonds. Moreover, synthetic PvD1 retained the biological activity against four different Candida species and showed no toxicity in-vivo. Adding the high resistance of synthetic PvD1 to proteolytic degradation we claim that conditions are now met to consider plant defensins druggable biologicals.
Julia Skalska; Vitor M. Andrade; Gabrielle L. Cena; Peta J. Harvey; Diana Maria Diez Gaspar; Érica O. Mello; Sónia Troeira Henriques; Javier Valle; Valdirene M. Gomes; Katia Conceição; Miguel A. R. B. Castanho; David Andreu. Synthesis, Structure, and Activity of the Antifungal Plant Defensin PvD1. Journal of Medicinal Chemistry 2020, 63, 1 .
AMA StyleJulia Skalska, Vitor M. Andrade, Gabrielle L. Cena, Peta J. Harvey, Diana Maria Diez Gaspar, Érica O. Mello, Sónia Troeira Henriques, Javier Valle, Valdirene M. Gomes, Katia Conceição, Miguel A. R. B. Castanho, David Andreu. Synthesis, Structure, and Activity of the Antifungal Plant Defensin PvD1. Journal of Medicinal Chemistry. 2020; 63 (17):1.
Chicago/Turabian StyleJulia Skalska; Vitor M. Andrade; Gabrielle L. Cena; Peta J. Harvey; Diana Maria Diez Gaspar; Érica O. Mello; Sónia Troeira Henriques; Javier Valle; Valdirene M. Gomes; Katia Conceição; Miguel A. R. B. Castanho; David Andreu. 2020. "Synthesis, Structure, and Activity of the Antifungal Plant Defensin PvD1." Journal of Medicinal Chemistry 63, no. 17: 1.
Clinical development of the COVID-19 vaccine candidate ChAdOx1 nCoV-19, a replication-deficient simian adenoviral vector expressing the full-length SARS-CoV-2 spike (S) protein was initiated in April 2020 following non-human primate studies using a single immunisation. Here, we compared the immunogenicity of one or two doses of ChAdOx1 nCoV-19 in both mice and pigs. Whilst a single dose induced antigen-specific antibody and T cells responses, a booster immunisation enhanced antibody responses, particularly in pigs, with a significant increase in SARS-CoV-2 neutralising titres.
Simon P. Graham; Rebecca K. McLean; Alexandra Spencer; Sandra Belij-Rammerstorfer; Daniel Wright; Marta Ulaszewska; Jane C. Edwards; Jack W. P. Hayes; Veronica Martini; Nazia Thakur; Carina Conceicao; Isabelle Dietrich; Holly Shelton; Ryan Waters; Anna Ludi; Ginette Wilsden; Clare Browning; Dagmara Bialy; Sushant Bhat; Phoebe Stevenson-Leggett; Philippa Hollinghurst; Ciaran Gilbride; David Pulido; Katy Moffat; Hannah Sharpe; Elizabeth Allen; Valerie Mioulet; Chris Chiu; Joseph Newman; Amin S. Asfor; Alison Burman; Sylvia Crossley; Jiandong Huo; Raymond J. Owens; Miles Carroll; John A. Hammond; Elma Tchilian; Dalan Bailey; Bryan Charleston; Sarah C. Gilbert; Tobias J. Tuthill; Teresa Lambe. Evaluation of the immunogenicity of prime-boost vaccination with the replication-deficient viral vectored COVID-19 vaccine candidate ChAdOx1 nCoV-19. npj Vaccines 2020, 5, 1 -6.
AMA StyleSimon P. Graham, Rebecca K. McLean, Alexandra Spencer, Sandra Belij-Rammerstorfer, Daniel Wright, Marta Ulaszewska, Jane C. Edwards, Jack W. P. Hayes, Veronica Martini, Nazia Thakur, Carina Conceicao, Isabelle Dietrich, Holly Shelton, Ryan Waters, Anna Ludi, Ginette Wilsden, Clare Browning, Dagmara Bialy, Sushant Bhat, Phoebe Stevenson-Leggett, Philippa Hollinghurst, Ciaran Gilbride, David Pulido, Katy Moffat, Hannah Sharpe, Elizabeth Allen, Valerie Mioulet, Chris Chiu, Joseph Newman, Amin S. Asfor, Alison Burman, Sylvia Crossley, Jiandong Huo, Raymond J. Owens, Miles Carroll, John A. Hammond, Elma Tchilian, Dalan Bailey, Bryan Charleston, Sarah C. Gilbert, Tobias J. Tuthill, Teresa Lambe. Evaluation of the immunogenicity of prime-boost vaccination with the replication-deficient viral vectored COVID-19 vaccine candidate ChAdOx1 nCoV-19. npj Vaccines. 2020; 5 (1):1-6.
Chicago/Turabian StyleSimon P. Graham; Rebecca K. McLean; Alexandra Spencer; Sandra Belij-Rammerstorfer; Daniel Wright; Marta Ulaszewska; Jane C. Edwards; Jack W. P. Hayes; Veronica Martini; Nazia Thakur; Carina Conceicao; Isabelle Dietrich; Holly Shelton; Ryan Waters; Anna Ludi; Ginette Wilsden; Clare Browning; Dagmara Bialy; Sushant Bhat; Phoebe Stevenson-Leggett; Philippa Hollinghurst; Ciaran Gilbride; David Pulido; Katy Moffat; Hannah Sharpe; Elizabeth Allen; Valerie Mioulet; Chris Chiu; Joseph Newman; Amin S. Asfor; Alison Burman; Sylvia Crossley; Jiandong Huo; Raymond J. Owens; Miles Carroll; John A. Hammond; Elma Tchilian; Dalan Bailey; Bryan Charleston; Sarah C. Gilbert; Tobias J. Tuthill; Teresa Lambe. 2020. "Evaluation of the immunogenicity of prime-boost vaccination with the replication-deficient viral vectored COVID-19 vaccine candidate ChAdOx1 nCoV-19." npj Vaccines 5, no. 1: 1-6.
A broadly protective and biosafe vaccine against foot-and-mouth disease virus (FMDV) remains an unmet need in the animal health sector. We have previously reported solid protection against serotype O FMDV afforded by dendrimeric peptide structures harboring virus-specific B- and T-cell epitopes, and also shown such type of multivalent presentations to be advantageous over simple B-T-epitope linear juxtaposition. Chemically, our vaccine platforms are modular constructions readily made from specified B- and T-cell epitope precursor peptides that are conjugated in solution. With the aim of developing an improved version of our formulations to be used for on-demand vaccine applications, we evaluate in this study a novel design for epitope presentation to the immune system based on a multiple antigen peptide (MAP) containing six immunologically relevant motifs arranged in dendrimeric fashion (named B2T-TB2). Interestingly, two B2T units fused tail-to-tail into a single homodimer platform elicited higher B- and T-cell specific responses than former candidates, with immunization scores remaining stable even after 4 months. Moreover, this macromolecular assembly shows consistent immune response in swine, the natural FMDV host, at reduced dose. Thus, our versatile, immunogenic prototype can find application in the development of peptide-based vaccine candidates for various therapeutic uses using safer and more efficacious vaccination regimens.
Sira Defaus; Mar Forner; Rodrigo Cañas-Arranz; Patricia De León; María J. Bustos; Miguel Rodríguez-Pulido; Esther Blanco; Francisco Sobrino; David Andreu. Designing Functionally Versatile, Highly Immunogenic Peptide-Based Multiepitopic Vaccines against Foot-and-Mouth Disease Virus. Vaccines 2020, 8, 406 .
AMA StyleSira Defaus, Mar Forner, Rodrigo Cañas-Arranz, Patricia De León, María J. Bustos, Miguel Rodríguez-Pulido, Esther Blanco, Francisco Sobrino, David Andreu. Designing Functionally Versatile, Highly Immunogenic Peptide-Based Multiepitopic Vaccines against Foot-and-Mouth Disease Virus. Vaccines. 2020; 8 (3):406.
Chicago/Turabian StyleSira Defaus; Mar Forner; Rodrigo Cañas-Arranz; Patricia De León; María J. Bustos; Miguel Rodríguez-Pulido; Esther Blanco; Francisco Sobrino; David Andreu. 2020. "Designing Functionally Versatile, Highly Immunogenic Peptide-Based Multiepitopic Vaccines against Foot-and-Mouth Disease Virus." Vaccines 8, no. 3: 406.