Assuntos
Fator B do Complemento , Fármacos Hematológicos , Hemoglobinúria Paroxística , Adulto , Feminino , Humanos , Masculino , Administração Oral , Fator B do Complemento/antagonistas & inibidores , Fármacos Hematológicos/administração & dosagem , Fármacos Hematológicos/efeitos adversos , Fármacos Hematológicos/uso terapêutico , Hemoglobinúria Paroxística/tratamento farmacológico , Hemoglobinúria Paroxística/complicaçõesAssuntos
Fator B do Complemento , Fármacos Hematológicos , Hemoglobinúria Paroxística , Humanos , Administração Oral , Fator B do Complemento/antagonistas & inibidores , Fármacos Hematológicos/administração & dosagem , Fármacos Hematológicos/efeitos adversos , Fármacos Hematológicos/uso terapêutico , Hemoglobinúria Paroxística/tratamento farmacológico , Hemoglobinúria Paroxística/complicaçõesAssuntos
Fator B do Complemento , Fármacos Hematológicos , Hemoglobinúria Paroxística , Adulto , Feminino , Humanos , Masculino , Administração Oral , Hemoglobinúria Paroxística/tratamento farmacológico , Hemoglobinúria Paroxística/complicações , Fator B do Complemento/antagonistas & inibidores , Fármacos Hematológicos/administração & dosagem , Fármacos Hematológicos/efeitos adversos , Fármacos Hematológicos/uso terapêuticoRESUMO
The complement system plays a crucial role in orchestrating the activation and regulation of inflammation within the human immune system. Three distinct activation pathways-classical, lectin, and alternative-converge to form the common lytic pathway, culminating in the formation of the membrane-attacking complex that disrupts the structure of pathogens. Dysregulated complement system activity can lead to tissue damage, autoimmune diseases, or immune deficiencies. In this study, the antimicrobial activity of human serum was investigated by using a bioluminescent microbe probe, Escherichia coli (pEGFPluxABCDEamp). This probe has previously been used to determine the antimicrobial activity of complement system and the polymorphonuclear neutrophils. In this study, blocking antibodies against key serum activators and components, including IgG, complement component 1q, factor B, and properdin, were utilized. The influence of body temperature and acute phase proteins, such as C reactive protein (CRP) and serum amyloid alpha (SAA), on the complement system was also examined. The study reveals the critical factors influencing complement system activity and pathway function. Alongside crucial factors like C1q and IgG, alternative pathway components factor B and properdin played pivotal roles. Results indicated that the alternative pathway accounted for approximately one third of the overall serum antimicrobial activity, and blocking this pathway disrupted the entire complement system. Contrary to expectations, elevated body temperature during inflammation did not enhance the antimicrobial activity of human serum. CRP demonstrated complement activation properties, but at higher physiological concentrations, it exhibited antagonistic tendencies, dampening the response. On the other hand, SAA enhanced the serum's activity. Overall, this study sheds a light on the critical factors affecting both complement system activity and pathway functionality, emphasizing the importance of a balanced immune response.
Assuntos
Temperatura Corporal , Proteína C-Reativa , Ativação do Complemento , Complemento C1q , Fator B do Complemento , Properdina , Proteína Amiloide A Sérica , Humanos , Proteína C-Reativa/metabolismo , Proteína C-Reativa/imunologia , Complemento C1q/imunologia , Complemento C1q/metabolismo , Proteína Amiloide A Sérica/metabolismo , Proteína Amiloide A Sérica/imunologia , Properdina/imunologia , Properdina/metabolismo , Fator B do Complemento/metabolismo , Fator B do Complemento/imunologia , Temperatura Corporal/imunologia , Escherichia coli/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologiaRESUMO
Iptacopan, the first orally available small-molecule complement factor B inhibitor, was developed by Novartis AG of Switzerland. Iptacopan for the treatment of PNH was just approved by the FDA in December 2023. Other indications for treatment are still in phase III clinical trials. Iptacopan is a small-molecule inhibitor targeting complement factor B, showing positive therapeutic effects in the treatment of PNH, C3 glomerulonephritis, and other diseases. Although Iptacopan is already on the market, there has been no detailed synthesis process or specific parameter report on the intermediates during the synthesis of its compounds except for the original research patent. In this study, a practical synthesis route for Iptacopan was obtained through incremental improvement while a biosynthesis method for ketoreductase was used for the synthesis of the pivotal intermediate 12. Moreover, by screening the existing enzyme library of our research group on the basis of random as well as site-directed mutagenesis methods, an enzyme (M8) proven to be of high optical purity with a high yield for biocatalectic reduction was obtained. This enzyme was used to prepare the compound benzyl (2S,4S)-4-hydroxy-2-(4-(methoxycarbonyl)-phenyl)-piperidine-1-carboxylate) white powder (36.8 g HPLC purity: 98%, ee value: 99%). In the synthesis of intermediate 15, the reaction was improved from two-step to one-step, which indicated that the risk of chiral allosterism was reduced while the scale was expanded. Finally, Iptacopan was synthesized in a seven-step reaction with a total yield of 29%. Since three chiral intermediate impurities were synthesized directionally, this paper lays a solid foundation for the future of pharmaceutical manufacturing.
Assuntos
Fator B do Complemento , Estrutura Molecular , Fator B do Complemento/antagonistas & inibidoresRESUMO
Paroxysmal nocturnal hemoglobinuria (PNH) is a rare, clonal, complement-mediated hemolytic anemia with a variety of manifestations. Currently, the methods for treating PNH include anti-C5 treatments (eculizumab and ravulizumab) and pegcetacoplan (a targeted C3 inhibitor). On December 5, 2023, the US FDA approved a factor B inhibitor called Fabhalta® (iptacopan), previously known as LNP023, for the treatment of adult patients with PNH, including those who have previously received anti-C5 therapy. The main objective of this review was to elucidate the clinical efficacy and safety of the newly approved factor B inhibitor, iptacopan. Iptacopan plays a proximal role in the alternative complement pathway to control extravascular hemolysis mediated by C3b and intravascular hemolysis mediated by terminal complement. The recommended dosage is 200 mg orally twice daily. The 24-week results of the pivotal phase III open-label trial, APPLY-PNH, demonstrated that among PNH patients who had previously received anti-C5 therapy, 51/60 (estimated percentages 82%) of patients in the iptacopan group showed an increase in hemoglobin of ≥2 g/dL compared to 0/35 (estimated percentages 2%) in the standard treatment group, also, 69% of iptacopan-treated patients achieved hemoglobin levels ≥12 g/dL, while no patients in the standard treatment group reached this level (both p < 0.001). The 48-week results were similar to those observed at 24 weeks. The most common adverse events were headache, infection and diarrhea. There were almost no clinical breakthrough hemolysis. Trials evaluating the long-term safety and efficacy of iptacopan are currently recruiting.
Assuntos
Hemoglobinúria Paroxística , Hemoglobinúria Paroxística/tratamento farmacológico , Humanos , Fator B do Complemento/antagonistas & inibidores , Resultado do Tratamento , Inativadores do Complemento/uso terapêutico , Inativadores do Complemento/efeitos adversosRESUMO
BACKGROUND: Numerous studies indicate a potential protective role of helminths in diabetes mellitus (DM) progression. The complement system, vital for host defense, plays a crucial role in tissue homeostasis and immune surveillance. Dysregulated complement activation is implicated in diabetic complications. We aimed to investigate the influence of the helminth, Strongyloides stercoralis (Ss) on complement activation in individuals with type 2 DM (T2D). METHODOLOGY: We assessed circulating levels of complement proteins (C1q, C2, C3, C4, C4b, C5, C5a, and MBL (Lectin)) and their regulatory components (Factor B, Factor D, Factor H, and Factor I) in individuals with T2D with (n = 60) or without concomitant Ss infection (n = 58). Additionally, we evaluated the impact of anthelmintic therapy on these parameters after 6 months in Ss-infected individuals (n = 60). RESULTS: Ss+DM+ individuals demonstrated reduced levels of complement proteins (C1q, C4b, MBL (Lectin), C3, C5a, and C3b/iC3b) and complement regulatory proteins (Factor B and Factor D) compared to Ss-DM+ individuals. Following anthelmintic therapy, there was a partial reversal of these levels in Ss+DM+ individuals. CONCLUSION: Our findings indicate that Ss infection reduces complement activation, potentially mitigating inflammatory processes in individuals with T2D. The study underscores the complex interplay between helminth infections, complement regulation, and diabetes mellitus, offering insights into potential therapeutic avenues.
Assuntos
Anti-Helmínticos , Diabetes Mellitus Tipo 2 , Helmintos , Strongyloides stercoralis , Estrongiloidíase , Animais , Humanos , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/tratamento farmacológico , Fator B do Complemento , Fator D do Complemento/uso terapêutico , Complemento C1q , Estrongiloidíase/complicações , Estrongiloidíase/tratamento farmacológico , Ativação do Complemento , Anti-Helmínticos/uso terapêutico , LectinasRESUMO
BACKGROUND: Persistent hemolytic anemia and a lack of oral treatments are challenges for patients with paroxysmal nocturnal hemoglobinuria who have received anti-C5 therapy or have not received complement inhibitors. Iptacopan, a first-in-class oral factor B inhibitor, has been shown to improve hemoglobin levels in these patients. METHODS: In two phase 3 trials, we assessed iptacopan monotherapy over a 24-week period in patients with hemoglobin levels of less than 10 g per deciliter. In the first, anti-C5-treated patients were randomly assigned to switch to iptacopan or to continue anti-C5 therapy. In the second, single-group trial, patients who had not received complement inhibitors and who had lactate dehydrogenase (LDH) levels more than 1.5 times the upper limit of the normal range received iptacopan. The two primary end points in the first trial were an increase in the hemoglobin level of at least 2 g per deciliter from baseline and a hemoglobin level of at least 12 g per deciliter, each without red-cell transfusion; the primary end point for the second trial was an increase in hemoglobin level of at least 2 g per deciliter from baseline without red-cell transfusion. RESULTS: In the first trial, 51 of the 60 patients who received iptacopan had an increase in the hemoglobin level of at least 2 g per deciliter from baseline, and 42 had a hemoglobin level of at least 12 g per deciliter, each without transfusion; none of the 35 anti-C5-treated patients attained the end-point levels. In the second trial, 31 of 33 patients had an increase in the hemoglobin level of at least 2 g per deciliter from baseline without red-cell transfusion. In the first trial, 59 of the 62 patients who received iptacopan and 14 of the 35 anti-C5-treated patients did not require or receive transfusion; in the second trial, no patients required or received transfusion. Treatment with iptacopan increased hemoglobin levels, reduced fatigue, reduced reticulocyte and bilirubin levels, and resulted in mean LDH levels that were less than 1.5 times the upper limit of the normal range. Headache was the most frequent adverse event with iptacopan. CONCLUSIONS: Iptacopan treatment improved hematologic and clinical outcomes in anti-C5-treated patients with persistent anemia - in whom iptacopan showed superiority to anti-C5 therapy - and in patients who had not received complement inhibitors. (Funded by Novartis; APPLY-PNH ClinicalTrials.gov number, NCT04558918; APPOINT-PNH ClinicalTrials.gov number, NCT04820530.).
Assuntos
Anemia Hemolítica , Fator B do Complemento , Inativadores do Complemento , Hemoglobinas , Hemoglobinúria Paroxística , Humanos , Administração Oral , Anemia Hemolítica/complicações , Complemento C5/antagonistas & inibidores , Fator B do Complemento/antagonistas & inibidores , Inativadores do Complemento/administração & dosagem , Inativadores do Complemento/efeitos adversos , Inativadores do Complemento/uso terapêutico , Transfusão de Eritrócitos , Cefaleia/induzido quimicamente , Hemoglobinas/análise , Hemoglobinúria Paroxística/tratamento farmacológico , Hemoglobinúria Paroxística/etiologia , Ensaios Clínicos Fase III como Assunto , Ensaios Clínicos Controlados Aleatórios como AssuntoRESUMO
This study aims to elucidate the cellular mechanisms behind the secretion of complement factor B (CFB), known for its dual roles as an early biomarker for pancreatic ductal adenocarcinoma (PDAC) and as the initial substrate for the alternative complement pathway (ACP). Using parallel reaction monitoring analysis, we confirmed a consistent â¼2-fold increase in CFB expression in PDAC patients compared with that in both healthy donors (HD) and chronic pancreatitis (CP) patients. Elevated ACP activity was observed in CP and other benign conditions compared with that in HD and PDAC patients, suggesting a functional link between ACP and PDAC. Protein-protein interaction analyses involving key complement proteins and their regulatory factors were conducted using blood samples from PDAC patients and cultured cell lines. Our findings revealed a complex control system governing the ACP and its regulatory factors, including Kirsten rat sarcoma viral oncogene homolog (KRAS) mutation, adrenomedullin (AM), and complement factor H (CFH). Particularly, AM emerged as a crucial player in CFB secretion, activating CFH and promoting its predominant binding to C3b over CFB. Mechanistically, our data suggest that the KRAS mutation stimulates AM expression, enhancing CFH activity in the fluid phase through binding. This heightened AM-CFH interaction conferred greater affinity for C3b over CFB, potentially suppressing the ACP cascade. This sequence of events likely culminated in the preferential release of ductal CFB into plasma during the early stages of PDAC. (Data set ID PXD047043.).
Assuntos
Carcinoma Ductal Pancreático , Neoplasias Pancreáticas , Humanos , Fator B do Complemento/genética , Fator B do Complemento/metabolismo , Via Alternativa do Complemento , Proteínas Proto-Oncogênicas p21(ras) , Detecção Precoce de Câncer , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/metabolismo , Carcinoma Ductal Pancreático/diagnóstico , Carcinoma Ductal Pancreático/genéticaRESUMO
Age-related macular degeneration (AMD) is the leading cause of irreversible vision loss among the elderly in the developed world. Whilst AMD is a multifactorial disease, the involvement of the complement system in its pathology is well documented, with single-nucleotide polymorphisms (SNPs) in different complement genes representing an increased risk factor. With several complement inhibitors explored in clinical trials showing limited success, patients with AMD are still without a reliable treatment option. This indicates that there is still a gap of knowledge in the functional implications and manipulation of the complement system in AMD, hindering the progress towards translational treatments. Since the discovery of the CRISPR/Cas system and its development into a powerful genome engineering tool, the field of molecular biology has been revolutionised. Genetic variants in the complement system have long been associated with an increased risk of AMD, and a variety of haplotypes have been identified to be predisposing/protective, with variation in complement genes believed to be the trigger for dysregulation of the cascade leading to inflammation. AMD-haplotypes (SNPs) alter specific aspects of the activation and regulation of the complement cascade, providing valuable insights into the pathogenic mechanisms of AMD with important diagnostic and therapeutic implications. The effect of targeting these AMD-related SNPs on the regulation of the complement cascade has been poorly explored, and the CRISPR/Cas system provides an ideal tool with which to explore this avenue. Current research concentrates on the association events of specific AMD-related SNPs in complement genes without looking into the effect of targeting these SNPs and therefore influencing the complement system in AMD pathogenesis. This review will explore the current understanding of manipulating the complement system in AMD pathogenesis utilising the genomic manipulation powers of the CRISPR/Cas systems. A number of AMD-related SNPs in different complement factor genes will be explored, with a particular emphasis on factor H (CFH), factor B (CFB), and complement C3 (C3).
Assuntos
Fator B do Complemento , Degeneração Macular , Humanos , Idoso , Haplótipos , Degeneração Macular/genética , Degeneração Macular/terapia , Degeneração Macular/patologia , Ativação do Complemento/genética , Fatores de Risco , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Mesangial complement C3 deposits, reflecting alternative and possibly lectin pathway activation, are characteristic in biopsies of patients with IgA nephropathy (IgAN). A recent randomized controlled trial tested the efficacy and safety of iptacopan, a factor B inhibitor, in patients with IgAN. Iptacopan dose-dependently reduced proteinuria, and there was a pronounced decrease of urinary C5b-9. This offers the perspective of "personalizing" therapy, which would be a unique feature of this novel approach to IgAN. A phase III clinical trial (APPLAUSE-IgAN) is ongoing.
Assuntos
Glomerulonefrite por IGA , Humanos , Glomerulonefrite por IGA/tratamento farmacológico , Ativação do Complemento , Mesângio Glomerular , Biópsia , Fator B do ComplementoRESUMO
Evaluating the activity of the complement system under conditions of altered thyroid hormone levels might help elucidate the role of complement in triggering autoimmune processes. Here, we investigated alternative pathway (AP) activity in male Wistar rats (180 ± 10 g) after altering their thyroid hormone levels by treatment with triiodothyronine (T3), propylthiouracil (PTU) or thyroidectomy. T3 and thyroxine (T4) levels were determined by chemiluminescence assays. Hemolytic assays were performed to evaluate the lytic activity of the AP. Factor B activity was evaluated using factor B-deficient serum. An anti-human factor B antibody was used to measure factor B levels in serum by radial immunodiffusion. T3 measurements in thyroidectomized animals or animals treated with PTU demonstrated a significant reduction in hormone levels compared to control. The results showed a reduction in AP lytic activity in rats treated with increasing amounts of T3 (1, 10, or 50 µg). Factor B activity was also decreased in the sera of hyperthyroid rats treated with 1 to 50 µg T3. Additionally, treating rats with 25 µg T3 significantly increased factor B levels in their sera (P < 0.01). In contrast, increased factor B concentration and activity (32 percent) were observed in hypothyroid rats. We conclude that alterations in thyroid hormone levels affect the activity of the AP and factor B, which may in turn affect the roles of AP and factor B in antibody production.
Assuntos
Animais , Masculino , Ratos , Antitireóideos/farmacologia , Fator B do Complemento/metabolismo , Via Alternativa do Complemento/efeitos dos fármacos , Propiltiouracila/farmacologia , Tiroxina/sangue , Tri-Iodotironina/sangue , Via Alternativa do Complemento/fisiologia , Hipertireoidismo/sangue , Hipertireoidismo/induzido quimicamente , Hipertireoidismo/imunologia , Hipotireoidismo/sangue , Hipotireoidismo/induzido quimicamente , Hipotireoidismo/imunologia , Medições Luminescentes , Ratos Wistar , TireoidectomiaRESUMO
OBJETIVO: Avaliar a atividade funcional das vias clássica e alternativa do sistema complemento e os níveis de C3, C4 e fator B durante o primeiro episódio de infecção meningocócica e durante a convalescença. PACIENTES E MÉTODOS: Dez crianças brasileiras com idades entre 8 meses e 8 anos, admitidas de 1991 a 1993, com diagnóstico clínico-laboratorial de meningite meningocócica, foram estudadas durante infecção aguda (até 7 dias do diagnóstico) e no período de convalescença (entre 1 e 6 meses após). C3, C4 e fator B foram quantificados por nefelometria e a atividade lítica das vias clássica e alternativa foi avaliada por método cinético e expressa como tempo necessário para lisar 50% de uma suspensão de eritrócitos (T1/2, expresso em segundos). Baixos valores de T1/2 das vias clássica e alternativa se correlacionam com elevadas atividades de via clássica e via alternativa, respectivamente. RESULTADOS: Observaram-se diferenças significativas entre a atividade lítica da via alternativa durante a infecção e no período de convalescença (282 e 238 segundos, respectivamente, P= .01). Nenhuma diferença foi detectada nos outros parâmetros analisados. CONCLUSÕES: Na presença de meningite meningocócica a via alternativa é preferencialmente ativada, provavelmente devido à maior capacidade da endotoxina meningocócica para ativar esta via, in vivo.
Assuntos
Humanos , Lactente , Pré-Escolar , Criança , Complemento C3 , Complemento C4 , Fator B do Complemento/análise , Via Alternativa do Complemento/imunologia , Via Clássica do Complemento/imunologia , Meningite Meningocócica/imunologia , Doença Aguda , Brasil , Ensaio de Atividade Hemolítica de Complemento , Convalescença , Meningite Meningocócica/sangue , Nefelometria e Turbidimetria , Valores de ReferênciaRESUMO
Objetivos - Analisar a variabilidade genética dos componentes C3 e BF do sistema complemento em pacientes brasileiros portadores de lúpus eritematoso sistêmico (LES) e as possíveis associações entre suas formas alotípicas e determinadas manifestações clínicas e laboratoriais da doença. Pacientes e métodos - O estudo foi realizado em 95 pacientes portadores de LES (88 mulheres e 7 homens, com variação etária de 14 a 57 anos, média de 30,18 anos), segundo os critérios de classificação do Colégio Americano de Reumatologia, e em 89 controles sadios. Os alótipos de C3 e de BF foram detectados no soro dos pacientes e controles através de eletroforese de alta voltagem em gel agarose, seguido de imunofixação com anticorpo específico. Resultados - Os alótipos de C3 e BF observados no presente estudo foram: C3S, C3F, C3SF, C3SS05 e BFS, BFF, BFSSF, BFSF1, BFSF075, BFSS07, BFF1. Os resultados obtidos demonstrarem aumento do alótipo BFF nos pacientes, quando comparados com os controles normais (p= 0,055; RR = 2,87); para os demais alótipos, não houve diferença signifiante quanto à sua distribuição. Menor frequência do alótipo BFS foi observada nos pacientes que apresentaram manifestações neurológicas, em relação aos que não as tiveram (p=0,059); RR = 0,28). Também nos pacientes que apresentaram serosites, observou-se frequência diminuída dos alótipos C3S e BFS, quando comparados com os que naõ apresentaram esta manifestação durante o curso da doença (p=0,036 para C3 e p=0,021 para BF; RR = 0,38 para ambos). Conclusões - A frequência diminuída de BFS nos pacientes com manifestações neurológicas e de C3S e BFS nos que apresentaram serosites no curso da doença sugere associação negativa e possível papel protetor desses alótipos no desenvolvimento dessas manifestações clínicas no LES. Os achados aqui descritos sugerem que a variabilidade genetica das proteínas C3 e BF do sistema complemento pode estar relacionada com o mecanismo etiopatogênico e com a expressão clínica do LES em pacientes brasileiros
Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto , Pessoa de Meia-Idade , Convertases de Complemento C3-C5 , Fator B do Complemento , Lúpus Eritematoso Sistêmico/etiologia , Polimorfismo GenéticoRESUMO
Se determinó la frecuencia de 29 antígenos HLA de los loci A y B en 20 pacientes con glaucoma primario de ángulo abierto diagnósticados en el Servicio de Oftalmología del Hospital General Docente ®Enrique Cabrera¼. Se utilizaron como controles 276 personas sanas. El antígeno HLA B35 mostró asociación positiva con un riesgo relativo (RR) de 5,3. Se obtuvo una frecuencia estadísticamente significativa con una p corregida (pc) <0,002 para el HLA B35 al compararla con controles normales no relacionados. El resto de los antígenos HLA estudiados no mostraron asociación
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Complexo Antígeno-Anticorpo , Complemento C3 , Complemento C4 , Fator B do Complemento , Via Alternativa do Complemento , Via Clássica do Complemento , Mieloma Múltiplo/imunologiaRESUMO
A variabilidade genética do fator B (BF) e do componente C3 do sistema complemento foi investigada numa populaçäo indígena Guarani do sul do Brasil. Foram analisados 70 indivíduos. As seguintes freqüências alélicas do loco BF foram observadas: BF*S=0,979, BF*F=0,014, BF*S05=0,007. A freqüência do alelo C3*S foi estimada em 100 por ciento. O alelo BF*S05 foi encontrado apenas em populaçöes sul-brasileiras, inclusive em índios Kaingang, o que indica que tenha se originado em indígenas sul-americanos. O baixo grau de polimorfismo de BF e o monomorfismo de C3, observado nos índios Guarani, estäo de acordo com os padröes de variabilidade observados em outras populaçöes ameríndias, esquimós e asiáticas.