Timing impact on the initiation of pirfenidone therapy on idiopathic pulmonary fibrosis disease progression

In this editorial,we comment on the article by Lei et al,with a specific focus on the timing of the initiation of the antifibrotic agent pirfenidone(PFD)in the management of idiopathic pulmonary fibrosis(IPF)and its impact on lung function of IPF patients.PFD is an antifibrotic agent that is widely used in the management of IPF in both early and advanced stages.It inhibits various pathways and has antifibrotic,anti-inflammatory,and antioxidant properties.Despite dosage lowering,PFD slowed IPF progression and maintained functional capacity.The 6-min walk distance test indicated that patients tolerated adverse events well,and PFD significantly reduced the incidence of progression episodes and death.Even when a single disease-progression event occurred,continuing PFD treatment had benefits.

Study on the efficacy of early treatment with pirfenidone on the lung function of patients with idiopathic pulmonary fibrosis

BACKGROUND Idiopathic pulmonary fibrosis(IPF)is classified under fibrotic interstitial pneumonia,characterized by a chronic and progressive course.The predominant clinical features of IPF include dyspnea and pulmonary dysfunction.AIM To assess the effects of pirfenidone in the early treatment of IPF on lung function in patients.METHODS A retrospective analysis was performed on 113 patients with IPF who were treated in our hospital from November 2017 to January 2023.These patients were divided into two groups:control group(n=53)and observation group(n=60).In the control group,patients received routine therapy in combination with methylprednisolone tablets,while those in the observation group received routine therapy together with pirfenidone.After applying these distinct treatment approaches to the two groups,we assessed several parameters,including the overall effectiveness of clinical therapy,the occurrence of adverse reactions(e.g.,nausea,vomiting,and anorexia),symptom severity scores,pulmonary function index levels,inflammatory marker levels,and the 6-min walk distance before and after treatment in both groups.RESULTS The observation group exhibited significantly higher rates than the control group after therapy,with a clear distinction(P<0.05).After treatment,the observation group experienced significantly fewer adverse reactions than the control group,with a noticeable difference(P<0.05).When analyzing the symptom severity scores between the two groups of patients after treatment,the observation group had significantly lower scores than the control group,with a distinct difference(P<0.05).When comparing the pulmonary function index levels between the two groups of patients after therapy,the observation group displayed significantly higher levels than the control group,with a noticeable difference(P<0.05).Evaluating the inflammatory marker data(C-reactive protein,interleukin-2[IL-2],and IL-8)between the two groups of patients after therapy,the observation group exhibited significantly lower levels than the control group,with significant disparities(P<0.05).Comparison of the 6-min walking distance data between the two groups of patients after treatment showed that the observation group achieved significantly greater distances than the control group,with a marked difference(P<0.05).CONCLUSION Prompt initiation of pirfenidone treatment in individuals diagnosed with IPF can enhance pulmonary function,elevate inflammatory factor levels,and increase the distance covered in the 6-min walk test.This intervention is conducive to effectively decreasing the occurrence of adverse reactions in patients.

Early pirfenidone treatment enhances lung function in idiopathic pulmonary fibrosis patients

This editorial comments on the study by Lei et al investigating the efficacy of early treatment with pirfenidone on the lung function of patients with idiopathic pulmonary fibrosis(IPF)published.This study evaluates the efficacy of early treatment with pirfenidone on lung function in patients with IPF.The early and advanced stages of IPF are defined,highlighting the drug's benefits.While prior research indicates pirfenidone's effectiveness in advanced IPF,this study focuses on its advantages in early stages.The study emphasizes the importance of computed tomography imaging alongside biochemical data and lung function tests for a comprehensive analysis of symptom relief.Results show that early intervention with pirfenidone significantly reduces disease progression and preserves lung function,underscoring its potential as a critical treatment strategy in early IPF.

Pirfenidone通过癌相关成纤维细胞抑制胆道肿瘤侵袭

目的探究吡非尼酮(PFD)通过癌相关成纤维细胞(CAF)抑制胆道肿瘤侵袭的相关机制。方法从胆道肿瘤患者的肿瘤组织中提取原代CAF,运用Western blot方法检测CAF的标志蛋白:波形蛋白(VIM),α-平滑肌肌动蛋白(α-SMA),成纤维细胞激活蛋白(FAP)。鬼笔环肽实验显示成纤维细胞骨架功能。ELISA和Western blot实验验证正常成纤维细胞(NF)与CAF的TGF-β表达差异。通过CAF中加入PFD观察CAF的功能变化。ELISA,实时荧光定量PCR(qRT-PCR)和Western blot实验验证CAF的TGF-β表达变化。通过皮下瘤小鼠模型验证血清中TGF-β的变化。胶原收缩实验观察CAF胶原收缩功能的改变。明胶酶谱实验观察CAF的培养基中的MMP2、MMP9的变化。Western blot方法检测CAF中SMAD信号通路蛋白变化。结果CAF的相关标志蛋白VIM,α-SMA,FAP均高表达,且CAF的丝状肌动蛋白(F-actin)表达丰富。ELISA实验显示CAF的TGF-β表达增强。Western blot实验验证CAF的胶原功能更强。Western blot实验、PCR实验和相关现象实验说明PFD能抑制CAF的胶原生成和TGF-β表达。SMAD信号通路相关蛋白实验证明PFD能通过抑制TGF-β/SMAD信号通路从而影响肿瘤侵袭。结论癌症患者中提取的CAF的功能以胶原生成为主,而PFD通过TGF-β/SMAD信号通路抑制CAF的胶原产生和胶原重塑相关过程从而起到抑制肿瘤侵袭的作用。

Explore potential biomarkers to provide a theoretical basis for the treatment of liver fibrosis with pirfenidone

Background:Hepatic fibrosis is a common chronic liver disease in clinic,the purpose of our study is to explore potential biomarkers to provide a theoretical basis for the treatment of liver fibrosis with pirfenidone.Methods:We downloaded a gene-sequencing dataset and a single-cell dataset from the GEO database and pirfenidone target genes from three different databases.First,we performed GO,KEGG,and DO analysis on pirfenidone target genes.Then,we grouped the liver tissue sequencing data(GSE162694)in the sequencing data set(N-F0 group and F1-F4 group)and performed gene expression differential analysis on these two groups,weighted gene co-expression network analysis and gene Enrichment analysis.Finally,we intersected the significantly upregulated genes in the F1-F4 group with the pirfenidone target genes and performed PPI network analysis.In order to further explore the expression of both pirfenidone drug target genes and liver fibrosis disease genes(PDLFG)in different immune cells of liver tissue,we used the CD45+cell data in the GSE136103 data set for further analysis.Results:A subnetwork consisting of CDC42,HNF4A,BHLHE40,CCDC71L,NR1H3,TNF,MGLL,GPT,SCD and PLIN1 was screened out,and by analysis,we finally identified the SCD as PDLFG.In single-cell sequencing analysis,we found that SCD was highly expressed in M2-polarized macrophages.Conclusion:SCD may be an important target protein to inhibit the progression of liver fibrosis.

Pirfenidone治疗肾、肺等组织纤维化的作用

随着对间质纤维化的细胞和生化机制的了解，新的治疗措施应针对防止间质纤维化的发生。Ｐｉｒｆｅｎｉｄｏｎｅ具有抗转化生长因子－β（ＴＧＦ－β）的作用。肾。

InterMune提交Pirfenidone的新药申请

InterMune公司已向FDA提交电子版新药申请（NDA），寻求批准pirfenidone（吡非尼酮）（Ⅰ）用于治疗特发性肺纤维化（IPF）。（Ⅰ）已被FDA认定为罕见病药物并给予快速审批资格，同时在欧洲也被认定为罕见病药物。

LOW DOSE PIRFENIDONE SUPPRESSES TRANSFORMING GROWTH FACTOR BETA-1 AND TISSUE INHIBITOR OF METALLOPROTEINASE-1, AND PROTECTS RATS FROM LUNG FIBROSIS INDUCED BY BLEOMYCIN

Objective To investigate the optimal dosage of pirfenidone for the treatment of pulmonary fibrosis induced by bleomycin in Wistar rats, and the alteration of expressions of transforming growth factor beta-1 （ TGF-β1 ）, tissue inhibitor of metalloproteinase-1 （ TIMP-1 ）, and matrix metalloproteinase-13 （ MMP-13 ） in lung tissue. Methods Male Wistar rats were endotracheally instilled with bleomycin or normal saline. Pirfenidone （25-800 mg · kg^-l · d^-1 ）, dexamethasone （3 mg/kg）, or 1% carboxymethylcellulose sodium were given daily by feed 2 days before instillation of bleomycin. Groups T7 and T14 were fed pirfenidone 50 mg · kg^-1 · d^-1 at 7 days or 14 daYs after bleomycin instillation. Lungs were harvested at 28 days after bleomycin instillation. Patholological changes in luffg tissues were evaluated with HE staining. Lung collagen was stained by sirius red and measured by content of hydroxypro- line. Expression of proteins of TGF-β1 TIMP-1, and MMP-13 were detected by Western blotting. Results At doses of 25, 50, and 100 mg· kg^- 1 · d ^- 1, pirfenidone had significant anti-fibrotic effects for bleomy- cin-induced rat pulmonary fibrosis, and these effects were most significantly attenuated at the dosage of 50 mg · kg^-1 ·d^ -1（ HE： P 〈 0. 01, P 〈 0.01, and P = 0.064; sirius red： P 〈0.05, P 〈 0.01, and P 〈 0.05 ; hydroxyproline： P = 0.595, P 〈 0.01, and P = 0.976）. Pirfenidone at a dosage of 50 mg · kg^- l · d^-1 inhibited protein expression of TGF-131 and TIMP-1 in lung tissue in the early phase （0.79 and 0.75 times of control group）, but had no effect on ex- nr^eelnn nf MMP-13. Conclusion Low dose pirfenidone, especially at dosage of 50 mg · kg^-1 · d^-1, has significant anti-fibrotic effects on bleomycin-induced rat pulmonary fibrosis. Pirfenidone partially inhibits the enhancement of the expression of TGF-131 and TIMP-β1 in lung tissue.

Pirfenidone suppresses the abnormal activation of human Müller cells after platelet-derived growth factor-BB stimulation

AIM: To determine the effect of pirfenidone on the activated human Müller cells by platelet-derived growth factor-BB(PDGF-BB). METHODS: The primary human Müller cells were separated from retinal tissues and established the pathogenic model by stimulated with PDGF-BB. The Müller cells behaviour of normal group and the model group was measured by MTT assay, Trypan blue assay, cell migration assay, and collagen contraction assay. The expression of transforming growth factor(TGF)-β1,-β2, and pigment epithelium-derived factor(PEDF) was estimated with realtime polymerase chain reaction(PCR), Western blot and immunofluorescence analyses. RESULTS: A pathogenic/proliferative model of Müller cells was established by stimulating normal cultured Müller cells with 10 ng/mL PDGF-BB for 48 h. After treated with 0.2 and 0.3 mg/mL pirfenidone, the proliferation, migration and collagen contraction was statistically significantly depressed in the model group compared with the normal groups. The expression levels of TGF-β1 and TGF-β2 were significantly down-regulated, while the PEDF expression was significantly up-regulated after treated with 0.2 and 0.3 mg/mL pirfenidone in the model group. CONCLUSION: Pirfenidone effectively suppress the proliferation, migration and collagen contraction of the human Müller cells stimulated with PDGF-BB through down-regulation of TGF-β1/TGF-β2 and up-regulation of PEDF.

Development and Substantiation of a RP-HPLC Method for Monitoring of Impurities in Pirfenidone Drug Substance

A simple, rapid and rugged RP-HPLC method was developed for evaluation and quantification of impurities present in Pirfenidone (PFD) drug substance. Impurities were separated and determined on a Zorbax RX-C18 column (250 mm length, 4.6 mm inner diameter and 5.0 μm particle size, octadecylsilane chemically bonded to porous silica) with 0.02 M KH2PO4 buffer and acetonitrile as mobile phase using a simple gradientelution program. The column flow rate of 1.0 mL per minute was used for the separation. The detection wave length was fixed at 220 nm. The method was substantiated with respect to specificity, precision, linearity, range, accuracy, ruggedness, limit of detection and quantitation. The impurities were identified as 2-hydroxy-5-methylpyridine and Iodobenzene. The linearity range obtained was 0.017 to 0.380 μg/mL for 2-hydroxy-5-methylpyridine, 0.047 to 0.382 μg/mL for Pirfenidone and 0.030 to 0.99 μg/mL for Iodobenzene with the retention times of 3.248 min, 10.608 min and 24.241 min for 2-hydroxy-5-methylpyridine, Pirfenidone and Iodobenzene, respectively. The percentage recoveries of 2-hydroxy-5-methylpyridine and Iodobenzene were in the range of 94.08% - 104.12%. The LOD and LOQ values were found 0.000005 mg/mL, 0.000017 mg/mL for 2-hydroxy-5-methylpyridine and 0.009 μg/mL, 0.030 μg/mL for Iodobenzene, respectively. The method is found to be suitable for the quantitation of impurities along with Pirfenidone drug substance. The method was validated as per the International Conference on Harmonization (ICH) guidelines.

Pirfenidone inhibits epithelial–mesenchymal transition in keloid keratinocytes

Background:Keloids are benign fibroproliferative skin lesions that are difficult to treat and become a lifetime predicament for patients.Several treatment modalities have been put forth,but as yet no satisfactory approach to the prevention or treatment of keloids has been identified.The process of epithelial-to-mesenchymal transition(EMT)has been implicated in keloid scarring,as keloid keratinocytes display an EMT-like phenotype.This study investigated the potential of pirfenidone,an antifibrotic agent,to counteract EMT-like alterations in keloid keratinocytes,including gene expression,cell migratory and proliferative functions.Methods:Normal and keloid keratinocytes were isolated from discarded normal skin tissues and from resected keloid tissues,respectively.Cells were quiesced for 24 h without epidermal growth factor DS-Qi1MCDigital and were exposed to transforming growth factor-beta1(TGF-β1;10 ng/mL),with or without pirfenidone(400μg/mL),for an additional 24 h.The effects of pirfenidone on cytotoxicity,cell migration,cell proliferation,and on expression of genes and proteins involved in EMT were assayed.Statistical significance was determined by two-way ANOVA using Sigma Plot.Results:We found that pirfenidone did not elicit any cytotoxic effect at concentrations up to 1000μg/mL.A statistically significant dose-dependent decrease in basal cell proliferation rate was noted in both normal and keloid keratinocytes when exposed to pirfenidone at concentrations ranging from 200 to 1000μg/mL.Pirfenidone significantly decreased basal cell migration in both normal and keloid keratinocytes,but a significant decrease in TGF-β1-induced cell migration was seen only in keloid keratinocytes.Significant inhibition of the expression of TGF-β1-induced core EMT genes,namely hyaluronan synthase 2,vimentin,cadherin-11,and wingless-type MMTV integration site family,member 5A along with fibronectin-1,was observed in both normal and keloid keratinocytes treated with pirfenidone.In addition,the protein levels of vimentin and fibronectin were significantly reduced by pirfenidone(400μg/mL)in both normal and keloid keratinocytes.Conclusions:For the first time,this study shows the efficacy of pirfenidone in inhibiting the EMTlike phenotype in keratinocytes derived from keloids,suggesting that pirfenidone may counteract a critical contributor of keloid progression and recurrence.

Repurposing Pirfenidone for Nonalcoholic Steatohepatitis-related Cirrhosis:A Case Series

We repurposed the antifibrotic drug pirfenidone—which is approved for treatment of idiopathic lung fibrosis—in a series of patients with nonalcoholic steatohepatitis-related cirrhosis.Our report demonstrates the observed improvements in necroinflammation and regression of cirrhosis with pirfeni-done use for 12-weeks,associated with classical hepatic repair complex features on follow-up liver biopsies.This novel work could help stimulate further randomized trials of pirfe-nidone in patients with nonalcoholic steatohepatitis-related liver fibrosis or cirrhosis,for whom no recommended drug treatments exists currently.

Effect of pirfenidone on renal tubulointerstitial fibrosis

Renal tubulointerstitialfibrosis(TIF)is the common end stage of various chronic renal diseases,and pirfenidone(PFD)is a novel,broad-spectrum anti-fibrotic compound but little is known about its effect and mechanism of action on renal TIF.In this work,we employed a unilateral ureteral obstruction(UUO)rat model to investigate the apoptosis of renal tubular epithelial cells(RTC)after PFD treatment.Thirty-five Sprague Dawley(SD)rats were randomized into three groups:sham-operated group(n=7),UUO group(n=14)and PFD group(n=14).All rats were sacrificed at day 7 or 14 after operation.Renal histology was studied by using periodic acid schiff reagent(PAS)and Masson trichromic stain(MASSON);apoptosis was detected by in situ terminal deoxynucleotide transferase-mediated dUTP-biotin nick end-labeling(TUNEL)method;tubular caspase-3 expression was assessed by immunohistochemistry.The content of malondialdehyde(MDA)and total activity of superoxide dismutase(T-SOD)in the renal cortex were determined by chemical colorimetry method.TIF,apopto-sis of RTC,tubular expression of caspase-3 and the content of MDA were increased in the UUO group compared with those in the sham-operated group,and were ameliorated significantly by PFD treatment(P<0.05).The activity of SOD was decreased in the UUO group,but was increased by PFD treatment(P<0.05).Our results showed that PFD could ameliorate TIF in the UUO group,and the possible mechanism was by reducing the apoptosis of RTC,which involved oxidative stress and caspase-3.