Antifibrotic activities of pirfenidone in animal models

C.J. Schaefer, D.W. Ruhrmund, L. Pan, S.D. Seiwert, K. Kossen
European Respiratory Review 2011 20: 85-97; DOI: 10.1183/09059180.00001111

Figures

  • Figure 1.
    Figure 1.

    Chemical structure of pirfenidone.

  • Figure 2.
    Figure 2.

    The antifibrotic activity of pirfenidone in preclinical models of pulmonary fibrosis. a) Experimental design and effects of prophylactic pirfenidone treatment on bleomycin-induced pulmonary fibrosis [38]. b) Effects of delayed pirfenidone treatment on bleomycin-induced pulmonary fibrosis (H. Oku, Shionogi and Co. Ltd, Osaka, Japan; personal communication). c) Effects of prophylactic pirfenidone treatment on bleomycin-induced pulmonary fibrosis [44]. d) Experimental design and effects of pirfenidone in an orthotopic lung transplant model [45]. e) The effects of pirfenidone in a mouse model of airway remodelling and hyperresponsiveness induced by sensitisation and challenge with ovalbumin [47]. BAL: bronchoalveolar lavage; bFGF: basic fibroblast growth factor; IL: interleukin; IFN: interferon; OVA: ovalbumin; PDGF: platelet-derived growth factor; SD: Sprague-Dawley; SDF: stromal cell-derived factor; TGF: transforming growth factor.

  • Figure 3.
    Figure 3.

    Summary of activities observed in animal models and cell-based assays. ECM: extracellular matrix.

Tables

  • Table 1. Effects of pirfenidone in animal models of lung, heart, kidney, and liver fibrosis
    SystemReferenceIn vivo modelSpeciesProtocolTotal daily dose/routeFibrosisFibrotic markersInflammatory markersOxidative stressFunctional end-points
    LungOku et al. [38]BleomycinMouseProphylactic10, 30*, 100* mg·kg−1 p.o.#DecreasedDecreasedDecreasedNANA
    H. Oku; personal communicationBleomycinMouseTherapeutic10, 30*, 100* mg·kg−1 p.o.#DecreasedNANANANA
    Kakugawa et al. [39]BleomycinMouseTherapeutic400* mg·kg−1 p.o.DecreasedDecreasedNANANA
    Iyer et al. [40]BleomycinHamsterProphylactic0.5%* in chowDecreasedDecreasedNADecreasedNA
    Iyer et al. [41]BleomycinHamsterTherapeutic+0.5%* in chowDecreasedDecreasedNADecreasedNA
    Iyer et al. [42]BleomycinHamsterProphylactic0.5%* in chowDecreasedDecreasedNADecreasedNA
    Gurujeyalakshmi et al. [43]BleomycinHamsterProphylactic0.5%* in chowNADecreasedNANANA
    Schelegle et al. [44]BleomycinHamsterProphylactic0.5%* in chowDecreasedNANANAImproved
    Liu et al. [45]TransplantRatProphylactic0.5%* in chowDecreasedDecreasedNANo effectImproved
    Zhou et al. [46]1) TransplantRatProphylactic0.5%* in chowDecreasedDecreasedNANANA
    2) TransplantRatTherapeutic0.5% in chowNo effectNANANANA
    Hirano et al. [47]Allergen induced dysfunctionMouseProphylactic125, 250, 500* mg·kg−1 s.c.§DecreasedDecreasedDecreasedNAImproved
    HeartLee et al. [2]Congestive heart failureDogProphylactic2400* mg p.o.#DecreasedDecreasedDecreasedNAImproved
    Nguyen et al. [3]MIRatTherapeutic1.2%* in chowDecreasedNANANAImproved
    Mirkovic et al. [33]HypertensionRatTherapeutic0.4%* in chowDecreasedNANANAImproved
    KidneyShimizu et al. [48]1) Continuous ureteral obstructionRatProphylactic0.6–0.9%* in chowDecreasedDecreasedNANANA
    2) Continuous ureteral obstructionRatTherapeutic0.6–0.9%ƒ in chowTrendDecreasedNANANA
    3) Temporary ureteral obstructionRatTherapeutic0.6–0.9%* in chowDecreasedNANANAImproved
    Takakura et al. [49]5/6 nephrectomyRatTherapeutic1%* in chowDecreasedDecreasedNANATrend
    Shimizu et al. [50]5/6 nephrectomyRatTherapeutic0.6–0.9%* in chowDecreasedDecreasedNANAImproved
    RamachandraRao et al. [51]Diabetic db/dbMouseTherapeutic0.5%* in chowDecreasedDecreasedNANANo effect
    LiverSalazar-Montes et al. [52]1) Carbon tetrachlorideRatTherapeutic200ƒ mg·kg−1 p.o.§TrendDecreasedNADecreasedImproved
    2) Bile duct ligationRatTherapeutic200* mg·kg−1 p.o.§DecreasedDecreasedNADecreasedNo effect
    Garcia et al. [53]1) Carbon tetrachlorideRatTherapeutic##500* mg·kg−1 p.o.DecreasedDecreasedNANAImproved
    2) Carbon tetrachlorideRatTherapeutic¶¶200* mg·kg−1 p.o.§DecreasedNANANANA
    3) Bile duct ligationRatProphylactic200*, 500 mg·kg−1 p.o.§DecreasedNANANANA
    Di Sario et al. [54]DimethylnitrosamineRatTherapeutic0.5%* in chowDecreasedDecreasedDecreasedNAImproved
    Tada et al. [55]DimethylnitrosamineRatProphylactic500* mg·kg−1 p.o.§DecreasedTrendNANANo effect

    • NA: not assessed; MI: myocardial infarction. *: statistically significant treatment effect (per reference); #: given in three divided daily doses; : given in two divided daily doses; +: pirfenidone treatment initiated following the second of three bleomycin insults; §: given in one daily dose; ƒ: treatment-associated trend; ##: pirfenidone dosed following cessation of carbon tetrachloride; ¶¶: pirfenidone dosed in final 3 weeks of an 11-week model with continuous carbon tetrachloride.

  • Table 2. Cytokines and growth factors modulated by pirfenidone treatment in animal models
    Cytokine/growth factorLungHeartKidneyLiver
    Oku et al. [38]Hirano et al. [47]Liu et al. [45]Zhou et al. [46]Lee et al. [2]Shimizu et al. [50]Shimizu et al. [48]Salazar-Montes et al. [52]Garcia et al. [53]Di Sario et al. [54]Tada et al. [55]
    BleomycinAllergen-induced AHRTransplantTransplantChronic heart failure5/6 nephrectomyUreteral obstructionCCl4 and bile duct ligation modelsCCl4 and bile duct ligation modelsDMNDMN
    TGF-βProtein↓Protein↓Protein↓Protein↓Protein↓mRNA↓mRNA↓mRNA↓mRNA↓mRNA↓mRNA↓#
    PDGFProtein↓
    bFGFProtein↓
    IL-18Protein↓
    SDF-1α/CXCL12Protein↓
    IFN-γProtein↔
    HGFmRNA↑
    MMP-2mRNA↓Protein↔mRNA↓
    MMP-9Protein↓
    TIMP-1mRNA↓mRNA↓
    TIMP-4Protein↑
    TNF-αProtein↓Protein↓
    IL-1βProtein↓
    IL-4Protein↓
    IL-5Protein↓
    IL-13Protein↓
    IL-6Protein↓
    MCP-1Protein↓
    IL-12p40Protein↓

    • Empty fields denote end-points that were either not assessed or not dysregulated in this model. Unless otherwise noted, all effects were statistically significant per publication. AHR: airway hyperresponsiveness; DMN: dimethylnitrosamine; TGF: transforming growth factor; PDGF: platelet-derived growth factor; bFGF: basic fibroblast growth factor; IL: interleukin; SDF: stromal cell-derived factor; CXCL12: chemokine, CXC motif, ligand 12; IFN: interferon; HGF: hepatocyte growth factor; MMP: matrix metalloproteinase; TIMP: tissue inhibitor of metalloproteinase; TNF: tumour necrosis factor; MCP: monocyte chemoattractant protein. #: statistical significance not assessed.

  • Table 3. Antifibrotic effects in cell-based assays
    ReferenceSystemFibroblast proliferationECM productionFibrotic markers
    Di Sario et al. [79]Primary rat hepatic stellate cellsInhibited PDGF-stimulated proliferationReduced TGF-β induced collagen I (protein and mRNA)
    Ozes and Blatt [80]Human lung fibroblasts (HFL-1)Reduced TGF-β induced collagen (protein)
    Sulfab et al. [81]Human unbilical vein endothelial cells (HUVEC)Reduced TGF-β stimulated fibronectin, collagen IV, and thrombospondin-2 proteins#Reduced TGF-β stimulated ET-1, IGFBP-3 and TGF-β isoforms 1-3
    Nakayama et al. [82]Normal human lung fibroblasts (NHLF)Reduced TGF-β induced collagen I expression (protein and mRNA)Reduced TGF-β induced expression of the collagen chaperone HSP47 (protein and mRNA)
    Zhang et al. [83]Human retinal pigment epithelial (RPE) cellsReduced TGF-β induced fibronectin (protein) production
    Hewitson et al. [84]Rat renal fibroblast isolated after ureteral obstruction (a mixture of fibroblasts and myofibroblasts)Reduced fibroblast proliferationCollagen protein production unaffected (slight reduction)Reduced α-SMA and CTGF protein expression
    Lin et al. [85]Human tenon’s fibroblasts (isolated from eye and expanded)Reduced serum-stimulated fibroblast proliferation. Increased population of G1-phase cells and decreased S-phase.Reduced TGF-β1, -β2, and -β3 mRNA and protein expression; reduced fibroblast migration and collagen matrix contraction
    Lee et al. [86]Human myometrial and leiomyoma smooth muscle cells (benign uterine fibroid tumour cells)Reduced serum-stimulated fibroblast proliferationReduced collagen I and III mRNA expression

    • Blank boxes denote end-points that were not assessed. All results statistically significant (per reference) with the exception of one treatment related trend (#). ECM: extracellular matrix; PDGF: platelet derived growth factor; TGF: transforming growth factor; ET: endothelin; IGFBP: insulin-like growth factor binding protein; HSP: heat shock protein; SMA: smooth muscle actin; CTGF: connective tissue growth factor; G1-phase: gap 1 phase; S-phase: synthesis phase.

  • Table 4. Anti-inflammatory effects in cell-based assays
    ReferenceSystemInflammatory markers
    Grattendick et al. [87]Human mononuclear cell line (THP-1)Reduced LPS-induced secreted TNF-α
    Reduced LPS-induced cell-associated TNF-α
    Phillips et al. [88]Human peripheral blood mononuclear cells (PBMC)Decreased LPS-induced TNF-α, IL-1β, IFN-γ and GM-CSF; increased LPS-stimulated IL-10
    Reduced constitutive expression of IL-8, IL-6, and MIP-1β
    Nakazato et al. [89]Mouse macrophage cell line (RAW264.7)Reduced LPS-induced secreted TNF-α
    Reduced LPS-induced cell-associated TNF-α

    • LPS: lipopolysaccharide; TNF: tumour necrosis factor; IL: interleukin; IFN: interferon; GM-CSF: granulocyte-macrophage colony-stimulating factor; MIP: macrophage inflammatory protein.

Additional Files

Back to top
View this article with LENS
Email
Print
Citation Tools

Antifibrotic activities of pirfenidone in animal models
C.J. Schaefer, D.W. Ruhrmund, L. Pan, S.D. Seiwert, K. Kossen
European Respiratory Review Jun 2011, 20 (120) 85-97; DOI: 10.1183/09059180.00001111
del.icio.us logo Digg logo Reddit logo Technorati logo Twitter logo CiteULike logo Connotea logo Facebook logo Google logo Mendeley logo
Full Text (PDF)

Jump To