RESUMEN
Acute lung injury (ALI) is a prevalent and severe clinical condition characterized by inflammatory damage to the lung endothelial and epithelial barriers, resulting in high incidence and mortality rates. Currently, there is a lack of safe and effective drugs for the treatment of ALI. In a previous clinical study, we observed that Jinyinqingre oral liquid (JYQR), a Traditional Chinese Medicine formulation prepared by the Taihe Hospital, Affiliated Hospital of Hubei University of Medicine, exhibited notable efficacy in treating inflammation-related hepatitis and cholecystitis in clinical settings. However, the potential role of JYQR in ALI/acute respiratory distress syndrome (ARDS) and its anti-inflammatory mechanism remains unexplored. Thus, the present study aimed to investigate the therapeutic effects and underlying molecular mechanisms of JYQR in ALI using a mouse model of lipopolysaccharide (LPS)-induced ALI and an in vitro RAW264.7 cell model. JYQR yielded substantial improvements in LPS-induced histological alterations in lung tissues. Additionally, JYQR administration led to a noteworthy reduction in total protein levels within the BALF, a decrease in MPAP, and attenuation of pleural thickness. These findings collectively highlight the remarkable efficacy of JYQR in mitigating the deleterious effects of LPS-induced ALI. Mechanistic investigations revealed that JYQR pretreatment significantly inhibited NF-κB activation and downregulated the expressions of the downstream proteins, namely NLRP3 and GSDMD, as well as proinflammatory cytokine levels in mice and RAW2647 cells. Consequently, JYQR alleviated LPS-induced ALI by inhibiting the NF-κB/NLRP3/GSDMD pathway. JYQR exerts a protective effect against LPS-induced ALI in mice, and its mechanism of action involves the downregulation of the NF-κB/NLRP3/GSDMD inflammatory pathway.
Asunto(s)
Lesión Pulmonar Aguda , FN-kappa B , Humanos , FN-kappa B/metabolismo , Lipopolisacáridos/toxicidad , Lipopolisacáridos/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Lesión Pulmonar Aguda/inducido químicamente , Lesión Pulmonar Aguda/tratamiento farmacológico , Lesión Pulmonar Aguda/metabolismo , Pulmón , Proteínas de Unión a Fosfato/metabolismo , Proteínas de Unión a Fosfato/farmacología , Proteínas de Unión a Fosfato/uso terapéutico , Proteínas Citotóxicas Formadoras de Poros/metabolismo , Proteínas Citotóxicas Formadoras de Poros/farmacología , Proteínas Citotóxicas Formadoras de Poros/uso terapéuticoRESUMEN
Antibiotic growth promoters (AGPs) have been administered in livestock for decades to improve food digestion in growing animals, while also contributing to the control of microbial pathogens. The long-term and indiscrimate use of AGPs has generated genetic modifications in bacteria, leading to antimicrobial resistance (AMR), which can be disseminated to commensal and pathogenic bacteria. Thus, antimicrobial peptides (AMPs) are used to replaced AGPs. AMPs are found in all domains of life, and their cationic characteristics can establish electrostatic interactions with the bacterial membrane. These molecules used as growth promoters can present benefits for nutrient digestibility, intestinal microbiota, intestinal morphology, and immune function activities. Therefore, this review focuses on the application of AMPs with growth promoting potential in livestock, as an alternative to conventional antibiotic growth promoters, in an attempt to control AMR. KEY POINTS: ⢠The long-term and indiscriminate use of AGPs in animal food can cause AMR. ⢠AMPs can be used as substitute of antibiotics in animal food suplementation. ⢠Animal food suplementated with AMPs can provied economic efficiency and sustainable livestock production.
Asunto(s)
Ganado , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Animales , Ganado/crecimiento & desarrolloRESUMEN
Curcumin (diferuloylmethane) is the main phytochemical of Curcuma longa Linn, an extract of the rhizome turmeric. For thousands of years, turmeric among other natural products has been used as a dietary spice and as a medicinal plant in Asian countries. The present study reports the leishmanicidal activity of curcumin in different concentrations (10 µM, 20 µM, 40 µM). It is also showing the effect of CM11 peptide (8 µM) alone and in combination with curcumin (10 and 20 µM) as a leishmanicidal drug. The experiments were performed with the amastigote form of Leishmania major (MRHO/IR/75/ER) in vitro and the leishmanicidal activity was analyzed after 12 and 24 h of incubation by Giemsa and DAPI staining. Further investigation was done by using semi-quantitative PCR with new designed common primer pair derived from an 18S rRNA gene belonging to the L. major and mouse, which amplified the above-mentioned gene segments simultaneously with different PCR product size. Our findings showed that curcumin had leishmanicidal activity in a dose and time-dependent manner and its lowest effective dose was at concentrations of 40 µM afetr12 h and 10 µM after 24 h. The IC50 value of curcumin against amastigote forms of L. major was 21.12 µM and 11.77 µM after 12 and 24 h, respectively. Treatment of amastigote form with CM11 (8 µM) alone and in combination with curcumin (10 µM and 20 µM) showed less leishmanicidal activity. Interestingly, CM11 in combination with curcumin (10 µM and 20 µM) had even less leishmanicidal effect compared to curcumin alone in the same concentrations (10 µM and 20 µM). The semi-quantitative PCR analysis confirmed the data achieved by Giemsa and DAPI staining and showed that curcumin reduced the PCR product derived from amastigote form in concentration and time-dependent manner compared to the genome of the host cells.
Asunto(s)
Péptidos Catiónicos Antimicrobianos/farmacología , Antiprotozoarios/farmacología , Curcumina/farmacología , Leishmania major/efectos de los fármacos , Leishmaniasis Cutánea/tratamiento farmacológico , Proteínas Citotóxicas Formadoras de Poros/farmacología , Análisis de Varianza , Animales , Péptidos Catiónicos Antimicrobianos/uso terapéutico , Antiprotozoarios/uso terapéutico , Curcumina/uso terapéutico , ADN Protozoario/química , ADN Protozoario/aislamiento & purificación , Relación Dosis-Respuesta a Droga , Concentración 50 Inhibidora , Irán , Leishmania major/genética , Leishmania major/crecimiento & desarrollo , Ratones , Reacción en Cadena de la Polimerasa , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Células RAW 264.7/parasitologíaRESUMEN
Anti-microbial peptides (AMPs), small biologically active molecules, produced by different organisms through their innate immune system, have become a considerable subject of interest in the request of novel therapeutics. Most of these peptides are cationic-amphipathic, exhibiting two main mechanisms of action, direct lysis and by modulating the immunity. The most commonly reported activity of AMPs is their anti-bacterial effects, although other effects, such as anti-fungal, anti-viral, and anti-parasitic, as well as anti-tumor mechanisms of action have also been described. Their anti-parasitic effect against leishmaniasis has been studied. Leishmaniasis is a neglected tropical disease. Currently among parasitic diseases, it is the second most threating illness after malaria. Clinical treatments, mainly antimonial derivatives, are related to drug resistance and some undesirable effects. Therefore, the development of new therapeutic agents has become a priority, and AMPs constitute a promising alternative. In this work, we describe the principal families of AMPs (melittin, cecropin, cathelicidin, defensin, magainin, temporin, dermaseptin, eumenitin, and histatin) exhibiting a potential anti-leishmanial activity, as well as their effectiveness against other microorganisms.
Asunto(s)
Antiprotozoarios/uso terapéutico , Leishmania/crecimiento & desarrollo , Leishmaniasis , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Animales , Humanos , Leishmaniasis/tratamiento farmacológico , Leishmaniasis/metabolismo , Leishmaniasis/patología , Malaria/tratamiento farmacológico , Malaria/metabolismo , Malaria/patologíaRESUMEN
The emergence of bacteria resistant to conventional antibiotics is of great concern in modern medicine because it renders ineffectiveness of the current empirical antibiotic therapies. Infections caused by vancomycin-resistant Staphylococcus aureus (VRSA) and vancomycin-intermediate S. aureus (VISA) strains represent a serious threat to global health due to their considerable morbidity and mortality rates. Therefore, there is an urgent need of research and development of new antimicrobial alternatives against these bacteria. In this context, the use of antimicrobial peptides (AMPs) is considered a promising alternative therapeutic strategy to control resistant strains. Therefore, a wide number of natural, artificial, and synthetic AMPs have been evaluated against VRSA and VISA strains, with great potential for clinical application. In this regard, we aimed to present a comprehensive and systematic review of research findings on AMPs that have shown antibacterial activity against vancomycin-resistant and vancomycin-intermediate resistant strains and clinical isolates of S. aureus, discussing their classification and origin, physicochemical and structural characteristics, and possible action mechanisms. This is the first review that includes all peptides that have shown antibacterial activity against VRSA and VISA strains exclusively.
Asunto(s)
Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Infecciones Estafilocócicas/tratamiento farmacológico , Staphylococcus aureus Resistente a Vancomicina/efectos de los fármacos , Humanos , Proteínas Citotóxicas Formadoras de Poros/química , Proteínas Citotóxicas Formadoras de Poros/clasificación , Proteínas Citotóxicas Formadoras de Poros/farmacologíaRESUMEN
The emergence of multidrug-resistant (MDR) bacteria through the abuse and long-term use of antibiotics is a serious health problem worldwide. Therefore, novel antimicrobial agents that can cure an infection from MDR bacteria, especially gram-negative bacteria, are urgently needed. Antimicrobial peptides, part of the innate immunity system, have been studied to find bactericidal agents potent against MDR bacteria. However, they have many problems, such as restrained systemic activity and cytotoxicity. In a previous study, we suggested that the K58-R78 domain of Romo1, a mitochondrial protein encoded by the nucleus, was a promising treatment candidate for sepsis caused by MDR bacteria. Here, we performed sequence optimization to enhance the antimicrobial activity of this peptide and named it as AMPR-22 (antimicrobial peptide derived from Romo1). It showed broad-spectrum antimicrobial activity against 17 sepsis-causing bacteria, including MDR strains, by inducing membrane permeabilization. Moreover, treatment with AMPR-22 enabled a remarkable survival rate in mice injected with MDR bacteria in a murine model of sepsis. Based on these results, we suggest that AMPR-22 could be prescribed as a first-line therapy (prior to bacterial identification) for patients diagnosed with sepsis.
Asunto(s)
Proteínas de la Membrana/química , Proteínas Mitocondriales/química , Fragmentos de Péptidos/uso terapéutico , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Sepsis/tratamiento farmacológico , Animales , Células Cultivadas , Farmacorresistencia Bacteriana , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Células HEK293 , Células HeLa , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Fragmentos de Péptidos/química , Fragmentos de Péptidos/farmacología , Proteínas Citotóxicas Formadoras de Poros/química , Proteínas Citotóxicas Formadoras de Poros/farmacología , Dominios Proteicos , Sepsis/microbiologíaRESUMEN
The objective of this study was to perform a comprehensive review of the use of antimicrobial peptides for the prevention and treatment of dental caries. The study included publications in the English language that addressed the use of antimicrobial peptides in the prevention and treatment of caries. These publications were also searchable on PubMed, Web of Science, Embase, Scopus, the Collection of Anti-Microbial Peptides and the Antimicrobial Peptide Database. A total of 3,436 publications were identified, and 67 publications were included. Eight publications reported seven natural human antimicrobial peptides as bactericidal to Streptococcus mutans. Fifty-nine publications reported 43 synthetic antimicrobial peptides developed to mimic natural antimicrobial peptides, fusing peptides with functional sequences and implementing new designs. The 43 synthetic antimicrobial peptides were effective against Streptococcus mutans, and nine peptides specifically targeted Streptococcus mutans. Ten antimicrobial peptides had an affinity for hydroxyapatite to prevent bacterial adhesion. Six antimicrobial peptides were also antifungal. Four antimicrobial peptides promoted remineralisation or prevented the demineralisation of teeth by binding calcium to hydroxyapatite. In conclusion, this study identified 67 works in the literature that reported seven natural and 43 synthetic antimicrobial peptides for the prevention and treatment of caries. Most of the antimicrobial peptides were bactericidal, and some prevented bacterial adhesion. A few antimicrobial peptides displayed remineralising properties with hydroxyapatite.
Asunto(s)
Antibacterianos/uso terapéutico , Caries Dental , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Adhesión Bacteriana/efectos de los fármacos , Biopelículas , Caries Dental/tratamiento farmacológico , Caries Dental/prevención & control , Durapatita , Humanos , Streptococcus mutans/efectos de los fármacos , Remineralización DentalRESUMEN
A serious pandemic has been caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The interaction between spike surface viral protein (Sgp) and the angiotensin-converting enzyme 2 (ACE2) cellular receptor is essential to understand the SARS-CoV-2 infectivity and pathogenicity. Currently, no drugs are available to treat the infection caused by this coronavirus and the use of antimicrobial peptides (AMPs) may be a promising alternative therapeutic strategy to control SARS-CoV-2. In this study, we investigated the in silico interaction of AMPs with viral structural proteins and host cell receptors. We screened the antimicrobial peptide database (APD3) and selected 15 peptides based on their physicochemical and antiviral properties. The interactions of AMPs with Sgp and ACE2 were performed by docking analysis. The results revealed that two amphibian AMPs, caerin 1.6 and caerin 1.10, had the highest affinity for Sgp proteins while interaction with the ACE2 receptor was reduced. The effective AMPs interacted particularly with Arg995 located in the S2 subunits of Sgp, which is key subunit that plays an essential role in viral fusion and entry into the host cell through ACE2. Given these computational findings, new potentially effective AMPs with antiviral properties for SARS-CoV-2 were identified, but they need experimental validation for their therapeutic effectiveness.
Asunto(s)
Enzima Convertidora de Angiotensina 2/química , Tratamiento Farmacológico de COVID-19 , SARS-CoV-2/química , Glicoproteína de la Espiga del Coronavirus/química , Proteínas Anfibias/química , Proteínas Anfibias/uso terapéutico , Enzima Convertidora de Angiotensina 2/genética , Enzima Convertidora de Angiotensina 2/uso terapéutico , Péptidos Catiónicos Antimicrobianos/química , Péptidos Catiónicos Antimicrobianos/uso terapéutico , Antivirales/química , Antivirales/uso terapéutico , Sitios de Unión/genética , COVID-19/genética , COVID-19/virología , Simulación por Computador , Humanos , Pandemias , Proteínas Citotóxicas Formadoras de Poros/química , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Unión Proteica/genética , SARS-CoV-2/patogenicidad , Glicoproteína de la Espiga del Coronavirus/uso terapéutico , Proteínas Estructurales Virales/química , Proteínas Estructurales Virales/genética , Proteínas Estructurales Virales/uso terapéuticoRESUMEN
At present, much attention is paid to the use of antimicrobial peptides (AMPs) of natural and artificial origin to combat pathogens. AMPs have several points that determine their biological activity. We analyzed the structural properties of AMPs, as well as described their mechanism of action and impact on pathogenic bacteria and viruses. Recently published data on the development of new AMP drugs based on a combination of molecular design and genetic engineering approaches are presented. In this article, we have focused on information on the amyloidogenic properties of AMP. This review examines AMP development strategies from the perspective of the current high prevalence of antibiotic-resistant bacteria, and the potential prospects and challenges of using AMPs against infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).
Asunto(s)
Proteínas Amiloidogénicas/farmacología , Tratamiento Farmacológico de COVID-19 , COVID-19/virología , Proteínas Citotóxicas Formadoras de Poros/farmacología , SARS-CoV-2/efectos de los fármacos , Proteínas Amiloidogénicas/efectos adversos , Proteínas Amiloidogénicas/uso terapéutico , Animales , Infecciones por Coronavirus/tratamiento farmacológico , Humanos , Proteínas Citotóxicas Formadoras de Poros/efectos adversos , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , ProteomaRESUMEN
Invasive fungal infections in humans are generally associated with high mortality, making the choice of antifungal drug crucial for the outcome of the patient. The limited spectrum of antifungals available and the development of drug resistance represent the main concerns for the current antifungal treatments, requiring alternative strategies. Antimicrobial peptides (AMPs), expressed in several organisms and used as first-line defenses against microbial infections, have emerged as potential candidates for developing new antifungal therapies, characterized by negligible host toxicity and low resistance rates. Most of the current literature focuses on peptides with antibacterial activity, but there are fewer studies of their antifungal properties. This review focuses on AMPs with antifungal effects, including their in vitro and in vivo activities, with the biological repercussions on the fungal cells, when known. The classification of the peptides is based on their mode of action: although the majority of AMPs exert their activity through the interaction with membranes, other mechanisms have been identified, including cell wall inhibition and nucleic acid binding. In addition, antifungal compounds with unknown modes of action are also described. The elucidation of such mechanisms can be useful to identify novel drug targets and, possibly, to serve as the templates for the synthesis of new antimicrobial compounds with increased activity and reduced host toxicity.
Asunto(s)
Péptidos Catiónicos Antimicrobianos/farmacología , Hongos/efectos de los fármacos , Proteínas Citotóxicas Formadoras de Poros/farmacología , Animales , Antifúngicos/química , Antifúngicos/uso terapéutico , Péptidos Catiónicos Antimicrobianos/biosíntesis , Péptidos Catiónicos Antimicrobianos/química , Péptidos Catiónicos Antimicrobianos/uso terapéutico , Vías Biosintéticas , Farmacorresistencia Fúngica , Humanos , Pruebas de Sensibilidad Microbiana , Micosis/tratamiento farmacológico , Micosis/microbiología , Proteínas Citotóxicas Formadoras de Poros/biosíntesis , Proteínas Citotóxicas Formadoras de Poros/química , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Relación Estructura-ActividadRESUMEN
Antimicrobial peptides (AMPs) are a potential alternative to classical antibiotics that are yet to achieve a therapeutic breakthrough for treatment of systemic infections. The antibacterial potency of pleurocidin, an AMP from Winter Flounder, is linked to its ability to cross bacterial plasma membranes and seek intracellular targets while also causing membrane damage. Here we describe modification strategies that generate pleurocidin analogues with substantially improved, broad spectrum, antibacterial properties, which are effective in murine models of bacterial lung infection. Increasing peptide-lipid intermolecular hydrogen bonding capabilities enhances conformational flexibility, associated with membrane translocation, but also membrane damage and potency, most notably against Gram-positive bacteria. This negates their ability to metabolically adapt to the AMP threat. An analogue comprising D-amino acids was well tolerated at an intravenous dose of 15 mg/kg and similarly effective as vancomycin in reducing EMRSA-15 lung CFU. This highlights the therapeutic potential of systemically delivered, bactericidal AMPs.
Asunto(s)
Antibacterianos/farmacología , Proteínas de Peces/farmacología , Enfermedades Pulmonares/tratamiento farmacológico , Proteínas Citotóxicas Formadoras de Poros/farmacología , Animales , Antibacterianos/química , Antibacterianos/uso terapéutico , Modelos Animales de Enfermedad , Proteínas de Peces/química , Proteínas de Peces/uso terapéutico , Células HEK293 , Células HeLa , Humanos , Enlace de Hidrógeno , Enfermedades Pulmonares/microbiología , Masculino , Membranas Artificiales , Ratones , Ratones Endogámicos C57BL , Pruebas de Sensibilidad Microbiana , Proteínas Citotóxicas Formadoras de Poros/química , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Conformación ProteicaRESUMEN
Novel antibiotics are urgently needed to combat multidrug-resistant pathogens. Venoms represent previously untapped sources of novel drugs. Here we repurposed mastoparan-L, the toxic active principle derived from the venom of the wasp Vespula lewisii, into synthetic antimicrobials. We engineered within its N terminus a motif conserved among natural peptides with potent immunomodulatory and antimicrobial activities. The resulting peptide, mast-MO, adopted an α-helical structure as determined by NMR, exhibited increased antibacterial properties comparable to standard-of-care antibiotics both in vitro and in vivo, and potentiated the activity of different classes of antibiotics. Mechanism-of-action studies revealed that mast-MO targets bacteria by rapidly permeabilizing their outer membrane. In animal models, the peptide displayed direct antimicrobial activity, led to enhanced ability to attract leukocytes to the infection site, and was able to control inflammation. Permutation studies depleted the remaining toxicity of mast-MO toward human cells, yielding derivatives with antiinfective activity in animals. We demonstrate a rational design strategy for repurposing venoms into promising antimicrobials.
Asunto(s)
Bacteriemia/tratamiento farmacológico , Proteínas Citotóxicas Formadoras de Poros/química , Venenos de Avispas/química , Animales , Diseño de Fármacos , Evaluación Preclínica de Medicamentos , Células HEK293 , Humanos , Ratones , Pruebas de Sensibilidad Microbiana , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Proteínas Citotóxicas Formadoras de Poros/toxicidad , Venenos de Avispas/uso terapéutico , Venenos de Avispas/toxicidadRESUMEN
The purpose of this review is to describe antifungal therapeutic candidates in preclinical and clinical development derived from, or directly influenced by, the immune system, with a specific focus on antimicrobial peptides (AMP). Although the focus of this review is AMP with direct antimicrobial effects on fungi, we will also discuss compounds with direct antifungal activity, including monoclonal antibodies (mAb), as well as immunomodulatory molecules that can enhance the immune response to fungal infection, including immunomodulatory AMP, vaccines, checkpoint inhibitors, interferon and colony stimulating factors as well as immune cell therapies. The focus of this manuscript will be a non-exhaustive review of antifungal compounds in preclinical and clinical development that are based on the principles of immunology and the authors acknowledge the incredible amount of in vitro and in vivo work that has been conducted to develop such therapeutic candidates.
Asunto(s)
Antifúngicos/uso terapéutico , Hongos/fisiología , Inmunoterapia/métodos , Micosis/terapia , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Animales , Ensayos Clínicos como Asunto , Evaluación Preclínica de Medicamentos , Humanos , Inmunidad InnataRESUMEN
Antimicrobial resistance is a multifaceted crisis, imposing a serious threat to global health. The traditional antibiotic pipeline has been exhausted, prompting research into alternate antimicrobial strategies. Inspired by nature, antimicrobial peptides are rapidly gaining attention for their clinical potential as they present distinct advantages over traditional antibiotics. Antimicrobial peptides are found in all forms of life and demonstrate a pivotal role in the innate immune system. Many antimicrobial peptides are evolutionarily conserved, with limited propensity for resistance. Additionally, chemical modifications to the peptide backbone can be used to improve biological activity and stability and reduce toxicity. This review details the therapeutic potential of peptide-based antimicrobials, as well as the challenges needed to overcome in order for clinical translation. We explore the proposed mechanisms of activity, design of synthetic biomimics, and how this novel class of antimicrobial compound may address the need for effective antibiotics. Finally, we discuss commercially available peptide-based antimicrobials and antimicrobial peptides in clinical trials.
Asunto(s)
Antibacterianos , Bacterias/efectos de los fármacos , Infecciones Bacterianas/tratamiento farmacológico , Diseño de Fármacos , Proteínas Citotóxicas Formadoras de Poros , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Humanos , Proteínas Citotóxicas Formadoras de Poros/farmacología , Proteínas Citotóxicas Formadoras de Poros/uso terapéuticoRESUMEN
A strategy was described to design antimicrobial peptides (AMPs) with enhanced salt resistance and antiendotoxin activities by linking two helical AMPs with the Ala-Gly-Pro (AGP) hinge. Among the designed peptides, KR12AGPWR6 demonstrated the best antimicrobial activities even in high salt conditions (NaCl ~300 mM) and possessed the strongest antiendotoxin activities. These activities may be related to hydrophobicity, membrane-permeability, and α-helical content of the peptide. Amino acids of the C-terminal helices were found to affect the peptide-induced permeabilization of LUVs, the α-helicity of the designed peptides under various LUVs, and the LPS aggregation and size alternation. A possible model was proposed to explain the mechanism of LPS neutralization by the designed peptides. These findings could provide a new approach for designing AMPs with enhanced salt resistance and antiendotoxin activities for potential therapeutic applications.
Asunto(s)
Endotoxemia/tratamiento farmacológico , Lipopolisacáridos/antagonistas & inhibidores , Proteínas Citotóxicas Formadoras de Poros/farmacología , Tolerancia a la Sal/efectos de los fármacos , Cloruro de Sodio/farmacología , Secuencia de Aminoácidos , Animales , Recuento de Colonia Microbiana , Evaluación Preclínica de Medicamentos , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Prueba de Limulus , Lipopolisacáridos/toxicidad , Masculino , Ratones , Ratones Endogámicos C57BL , Proteínas Citotóxicas Formadoras de Poros/síntesis química , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Conformación Proteica en Hélice alfa , Relación Estructura-Actividad , Factor de Necrosis Tumoral alfa/sangre , Liposomas UnilamelaresRESUMEN
Muramic acid (Mur), a sugar amino acid (SAA), is present in the cell walls of bacteria asN-acetyl muramic acid (MurNAc) where together with ofN-acetylglucosamine (GlcNAc) and peptide makes main building block of peptidoglycan (PGN). It was challenging to incorporate muramic acid as SAA characteristic for bacteria into the peptides and investigate the antimicrobial activity of these scaffolds. Four building units were used in designing the desired peptide: muramic acid, tetrapeptide Leu-Ser-Lys-Leu, Nε-Lys, and Asn. Positions of three components were changeable while the position of Asn was always C-terminal (in linear peptides). The glycopeptide libraries of linear and cyclic peptides were synthesized using solid-phase peptide synthesis (SPPS). The antimicrobial effect of linear and cyclic glycopeptides, as well as the LSKL sequence used as a control, was investigated on several standard laboratory microbial strains. Liner glycopeptide with sequences Leu-Ser-Lys-Leu-Nε-Lys-Mur-Asn was active onStaphylococcus aureus(Gram-positive bacteria). Prepared compounds did not show activity towards applied tumor and normal human cell lines.
Asunto(s)
Ácidos Murámicos/uso terapéutico , Proteínas Citotóxicas Formadoras de Poros/síntesis química , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Humanos , Estructura Molecular , Ácidos Murámicos/farmacología , Proteínas Citotóxicas Formadoras de Poros/farmacología , Relación Estructura-ActividadRESUMEN
Bombyx mori antimicrobial peptides (BmAMPs) are important effectors in silkworm immune system. They can inhibit and kill a variety of bacteria and fungi. Recent studies have shown that some kinds of BmAMPs exert strong inhibitory effects on a variety of tumor cells. In the present study, the antitumor activity of BmAMP Cecropin A (BmCecA) and BmAMP Cecropin D (BmCecD) was investigated against human esophageal cancer cells and their antitumor mechanism preliminary explored. Cell Counting Kit-8 and colony formation assays indicated that BmCecA and BmCecD suppressed cell proliferation and reduced colony formation of both Eca109 and TE13 cells in a dose-dependent manner, but exhibited no inhibitory effect on normal human embryonic kidney 293T cells. Wound healing and invasion experiments indicated that both BmCecA and BmCecD inhibited migration and invasion of Eca109 and TE13 cells in vitro. Annexin V/propidium iodide staining and flow cytometry detection suggested that BmCecA induced the apoptosis of Eca109 cells in a dose-dependent manner. RT-qPCR and western blot analysis showed that BmCecA induced apoptosis of Eca109 cells through the activation of a mitochondria-mediated caspase pathway, the upregulation of B-cell lymphoma 2 (Bcl-2)-associated X protein and the downregulation of Bcl-2. In addition, BmCecA significantly inhibited the growth of xenograft tumors in Eca109-bearing mice. These results suggested that BmCecA and BmCecD might serve as potential therapeutic agents for the treatment of cancer in the future.
Asunto(s)
Bombyx , Cecropinas/uso terapéutico , Neoplasias Esofágicas/prevención & control , Proteínas Citotóxicas Formadoras de Poros/uso terapéutico , Secuencia de Aminoácidos , Animales , Apoptosis/efectos de los fármacos , Apoptosis/fisiología , Cecropinas/aislamiento & purificación , Cecropinas/farmacología , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Movimiento Celular/fisiología , Neoplasias Esofágicas/patología , Femenino , Células HEK293 , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Proteínas Citotóxicas Formadoras de Poros/aislamiento & purificación , Proteínas Citotóxicas Formadoras de Poros/farmacologíaRESUMEN
The rapid development of microbial resistance to conventional antibiotics has accelerated efforts to find anti-infectives with a novel mode-of-action, which are less prone to bacterial resistance. Intense nonclinical and clinical research is today ongoing to evaluate antimicrobial peptides (AMPs) as potential next-generation antibiotics. Currently, multiple AMPs are assessed in late-stage clinical trials, not only as novel anti-infective drugs, but also as innovative product candidates for immunomodulation, promotion of wound healing, and prevention of post-operative scars. The efforts to translate AMP-based research findings into pharmaceutical product candidates are expected to accelerate in coming years due to technological advancements in multiple areas, including an improved understanding of the mechanism-of-action of AMPs, smart formulation strategies, and advanced chemical synthesis protocols. At the same time, it is recognized that cytotoxicity, low metabolic stability due to sensitivity to proteolytic degradation, and limited oral bioavailability are some of the key weaknesses of AMPs. Furthermore, the pricing and reimbursement environment for new antimicrobial products remains as a major barrier to the commercialization of AMPs.
Asunto(s)
Proteínas Citotóxicas Formadoras de Poros , Animales , Antiinfecciosos/uso terapéutico , Bacterias/efectos de los fármacos , Infecciones Bacterianas/tratamiento farmacológico , Farmacorresistencia Bacteriana , Humanos , Ratones , Proteínas Citotóxicas Formadoras de Poros/química , Proteínas Citotóxicas Formadoras de Poros/farmacología , Proteínas Citotóxicas Formadoras de Poros/uso terapéuticoRESUMEN
BACKGROUND: Antimicrobial peptide derived from insulin-like growth factor binding protein-5 (AMP-IBP5) is a potent antimicrobial agent that possesses various immunomodulatory activities. The parent protein of AMP-IBP5, IGFBP-5, has been shown to exert its effects via an insulin-like growth factor-1 receptor-independent mechanism, including binding to multifunctional low-density lipoprotein receptor-related protein 1 (LRP1), which contributes to several biological processes involved in skin wound healing. OBJECTIVES: To investigate whether LRP1 is involved in AMP-IBP5-induced migration and proliferation of human epidermal keratinocytes and dermal fibroblasts. METHODS: The mRNA expression of LRP1 and IGFBP-5 was assessed by quantitative real-time PCR, whereas Western blotting was used to evaluate the protein expression. Production of cytokines was determined by ELISA. Cell migration was measured by the scratch wound assay, whereas cell proliferation was analyzed using the BrdU labeling assay. MAPK activation was determined by Western blotting. RESULTS: We found that AMP-IBP5 markedly induced the migration and proliferation of keratinocytes and fibroblasts, and this effect was reversed by specific siRNA and neutralizing antibody targeting the LRP1 receptor. In addition, LRP1 was upregulated by lipopolysaccharide, flagellin and AMP-IBP5 in keratinocytes and fibroblasts. LRP1 knockdown also inhibited MAPK pathway activation, which was required for AMP-IBP5-mediated cell migration and proliferation, as evidenced by the specific inhibitors for extracellular signal-regulated kinase, c-Jun N-terminal kinase and p38. CONCLUSIONS: Our results suggest that LRP1 expressed in human epidermal keratinocytes and dermal fibroblasts contributes to AMP-IBP5-mediated cell migration and proliferation, supporting its crucial role in cutaneous wound healing process.