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Diacetyloxyl derivatization of the fibroblast growth factor inhibitor dobesilate enhances its anti-inflammatory, anti-angiogenic and anti-tumoral activities.

Angulo J, Cuevas P, Cuevas B, El Youssef M, Fernández A, Martínez-Salamanca E, González-Corrochano R, Giménez-Gallego G - J Transl Med (2015)

Bottom Line: Topical DAPS is more effective than DHPS in preventing inflammatory signs (increased vascular permeability, edema, leukocyte infiltration, MPO activation) caused by contact dermatitis induction in rat ears.DAPS, but not DHPS, effectively inhibits COX-1 and COX-2 activities.DAPS also reduces the increase in serum cytokine concentration induced by lipopolysaccharide in rats.

View Article: PubMed Central - PubMed

Affiliation: Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Hospital Universitario Ramón y Cajal, Madrid, Spain. javier.angulo@hrc.es.

ABSTRACT

Background: Dobesilate (2,5-dihydroxyphenyl sulfonate, DHPS) was recently identified as the most potent member of a family of fibroblast growth factor (FGF) inhibitors headed by gentisic acid, one of the main catabolites of aspirin. Although FGFs were first described as inducers of angiogenesis, they were soon recognized as broad spectrum mitogens. Furthermore, in the last decade these proteins have been shown to participate directly in the onset of inflammation, and their potential angiogenic activity often contributes to the inflammatory process in vivo. The aim of this work was to evaluate the anti-inflammatory, anti-angiogenic and anti-tumoral activities of the derivative of DHPS obtained by acetoxylation of its two hydroxyl groups (2,5-diacetoxyphenyl sulfonate; DAPS).

Methods: Anti-inflammatory, anti-angiogenic and anti-tumoral activities of DHPS and DAPS were compared using in vivo assays of dermatitis, angiogenesis and tumorigenesis. The effects of both compounds on myeloperoxidase (MPO) and cyclooxygenase (COX) activities, cytokine production and FGF-induced fibroblast proliferation were also determined.

Results: Topical DAPS is more effective than DHPS in preventing inflammatory signs (increased vascular permeability, edema, leukocyte infiltration, MPO activation) caused by contact dermatitis induction in rat ears. DAPS, but not DHPS, effectively inhibits COX-1 and COX-2 activities. DAPS also reduces the increase in serum cytokine concentration induced by lipopolysaccharide in rats. Furthermore, DAPS displays higher in vivo efficacy than DHPS in inhibiting FGF-induced angiogenesis and heterotopic glioma progression, with demonstrated oral efficacy to combat both processes.

Conclusions: By inhibiting both FGF-signaling and COX-mediated prostaglandin synthesis, DAPS efficiently breaks the vicious circle created by the reciprocal induction of FGF and prostaglandins, which probably sustains undesirable inflammation in many circumstances. Our findings define the enhancement of anti-inflammatory, anti-angiogenic and anti-tumoral activities by diacetyloxyl derivatization of the FGF inhibitor, dobesilate.

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Related in: MedlinePlus

Representative histology showing the effects of topical treatment with 2,5-diacetoxyphenyl sulfonate (DAPS; 2.5%; eq. to 0.08 mmol/ml) on benzalkonium chloride-induced dermatitis in the rat ear. The first row shows the macroscopic appearance of both ears of a rat treated with vehicle alone (glycerol: A) and those of a rat treated with DAPS (B), which evidently reduces the erythema caused by dermatitis. In the second row, the tissue edema on the ear of a rat with dermatitis treated only with glycerol (C) was not observed in DAPS-treated rats (D). The third row shows the intense leukocyte infiltration in a glycerol-treated rat ear (E), an effect that was clearly reduced by DAPS treatment (F). Magnification of the boxed area in E and F reveals that in the capillaries of vehicle-treated rats, there are leukocytes adhered to the endothelial cells, which rolled and extravasated to infiltrate the surrounding tissue (G), a feature not observed in the vessels of DAPS-treated rats (H). The infiltration of leukocytes into the erector muscle of the ear in glycerol-treated rats (I) was also attenuated by DAPS treatment (J). Magnifications: C-F, x100; G-H x400; I-J x200. Panel K shows the effects of topic treatment with 2,5-dihydroxyphenyl sulfonate (DHPS; 5%; eq. to 0.22 mmol/ml), DAPS (2.5%; eq. to 0.08 mmol/ml) or the vehicle alone (glycerol) on the myeloperoxidase (MPO) activity associated with benzalkonium chloride-induced dermatitis on the rat ear. Dermatitis was not induced in the control group. MPO activity is expressed as the mean ± SEM of the absorbance at 460 nm normalized to the weight (mg) of the tissue of the corresponding ear. The numbers of animals used is shown in parentheses: **p < 0.01 vs. control; † p < 0.05, †† p < 0.01 vs. vehicle by one-factor ANOVA followed by Student-Newmann-Keuls test.
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Fig2: Representative histology showing the effects of topical treatment with 2,5-diacetoxyphenyl sulfonate (DAPS; 2.5%; eq. to 0.08 mmol/ml) on benzalkonium chloride-induced dermatitis in the rat ear. The first row shows the macroscopic appearance of both ears of a rat treated with vehicle alone (glycerol: A) and those of a rat treated with DAPS (B), which evidently reduces the erythema caused by dermatitis. In the second row, the tissue edema on the ear of a rat with dermatitis treated only with glycerol (C) was not observed in DAPS-treated rats (D). The third row shows the intense leukocyte infiltration in a glycerol-treated rat ear (E), an effect that was clearly reduced by DAPS treatment (F). Magnification of the boxed area in E and F reveals that in the capillaries of vehicle-treated rats, there are leukocytes adhered to the endothelial cells, which rolled and extravasated to infiltrate the surrounding tissue (G), a feature not observed in the vessels of DAPS-treated rats (H). The infiltration of leukocytes into the erector muscle of the ear in glycerol-treated rats (I) was also attenuated by DAPS treatment (J). Magnifications: C-F, x100; G-H x400; I-J x200. Panel K shows the effects of topic treatment with 2,5-dihydroxyphenyl sulfonate (DHPS; 5%; eq. to 0.22 mmol/ml), DAPS (2.5%; eq. to 0.08 mmol/ml) or the vehicle alone (glycerol) on the myeloperoxidase (MPO) activity associated with benzalkonium chloride-induced dermatitis on the rat ear. Dermatitis was not induced in the control group. MPO activity is expressed as the mean ± SEM of the absorbance at 460 nm normalized to the weight (mg) of the tissue of the corresponding ear. The numbers of animals used is shown in parentheses: **p < 0.01 vs. control; † p < 0.05, †† p < 0.01 vs. vehicle by one-factor ANOVA followed by Student-Newmann-Keuls test.

Mentions: In the absence of dye administration, a marked erythematous response was observed 24 h after the induction of dermatitis with 5% BZK in rat ears (Figure 2A) which was notably prevented by topical application of DAPS (2.5%) (Figure 2B). A detailed histological characterization of the inflammatory response confirmed that topic application of DAPS (2.5%) reduced the extensive skin edema (Figure 2C-D) and leukocyte infiltration (Figure 2E-H) associated with dermatitis in untreated animals. Dermatomyositis was also observed in the erector muscle of the ear after BZK-induced dermatitis (Figure 2I) and likewise, it was prevented by DAPS administration (Figure 2J). The involvement of leukocyte infiltration in this inflammatory response was confirmed by the significant elevation of MPO activity. This increase was prevented by topical administration of DHPS (5%), and more effectively with 2.5% DAPS (Figure 2K).Figure 2


Diacetyloxyl derivatization of the fibroblast growth factor inhibitor dobesilate enhances its anti-inflammatory, anti-angiogenic and anti-tumoral activities.

Angulo J, Cuevas P, Cuevas B, El Youssef M, Fernández A, Martínez-Salamanca E, González-Corrochano R, Giménez-Gallego G - J Transl Med (2015)

Representative histology showing the effects of topical treatment with 2,5-diacetoxyphenyl sulfonate (DAPS; 2.5%; eq. to 0.08 mmol/ml) on benzalkonium chloride-induced dermatitis in the rat ear. The first row shows the macroscopic appearance of both ears of a rat treated with vehicle alone (glycerol: A) and those of a rat treated with DAPS (B), which evidently reduces the erythema caused by dermatitis. In the second row, the tissue edema on the ear of a rat with dermatitis treated only with glycerol (C) was not observed in DAPS-treated rats (D). The third row shows the intense leukocyte infiltration in a glycerol-treated rat ear (E), an effect that was clearly reduced by DAPS treatment (F). Magnification of the boxed area in E and F reveals that in the capillaries of vehicle-treated rats, there are leukocytes adhered to the endothelial cells, which rolled and extravasated to infiltrate the surrounding tissue (G), a feature not observed in the vessels of DAPS-treated rats (H). The infiltration of leukocytes into the erector muscle of the ear in glycerol-treated rats (I) was also attenuated by DAPS treatment (J). Magnifications: C-F, x100; G-H x400; I-J x200. Panel K shows the effects of topic treatment with 2,5-dihydroxyphenyl sulfonate (DHPS; 5%; eq. to 0.22 mmol/ml), DAPS (2.5%; eq. to 0.08 mmol/ml) or the vehicle alone (glycerol) on the myeloperoxidase (MPO) activity associated with benzalkonium chloride-induced dermatitis on the rat ear. Dermatitis was not induced in the control group. MPO activity is expressed as the mean ± SEM of the absorbance at 460 nm normalized to the weight (mg) of the tissue of the corresponding ear. The numbers of animals used is shown in parentheses: **p < 0.01 vs. control; † p < 0.05, †† p < 0.01 vs. vehicle by one-factor ANOVA followed by Student-Newmann-Keuls test.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Fig2: Representative histology showing the effects of topical treatment with 2,5-diacetoxyphenyl sulfonate (DAPS; 2.5%; eq. to 0.08 mmol/ml) on benzalkonium chloride-induced dermatitis in the rat ear. The first row shows the macroscopic appearance of both ears of a rat treated with vehicle alone (glycerol: A) and those of a rat treated with DAPS (B), which evidently reduces the erythema caused by dermatitis. In the second row, the tissue edema on the ear of a rat with dermatitis treated only with glycerol (C) was not observed in DAPS-treated rats (D). The third row shows the intense leukocyte infiltration in a glycerol-treated rat ear (E), an effect that was clearly reduced by DAPS treatment (F). Magnification of the boxed area in E and F reveals that in the capillaries of vehicle-treated rats, there are leukocytes adhered to the endothelial cells, which rolled and extravasated to infiltrate the surrounding tissue (G), a feature not observed in the vessels of DAPS-treated rats (H). The infiltration of leukocytes into the erector muscle of the ear in glycerol-treated rats (I) was also attenuated by DAPS treatment (J). Magnifications: C-F, x100; G-H x400; I-J x200. Panel K shows the effects of topic treatment with 2,5-dihydroxyphenyl sulfonate (DHPS; 5%; eq. to 0.22 mmol/ml), DAPS (2.5%; eq. to 0.08 mmol/ml) or the vehicle alone (glycerol) on the myeloperoxidase (MPO) activity associated with benzalkonium chloride-induced dermatitis on the rat ear. Dermatitis was not induced in the control group. MPO activity is expressed as the mean ± SEM of the absorbance at 460 nm normalized to the weight (mg) of the tissue of the corresponding ear. The numbers of animals used is shown in parentheses: **p < 0.01 vs. control; † p < 0.05, †† p < 0.01 vs. vehicle by one-factor ANOVA followed by Student-Newmann-Keuls test.
Mentions: In the absence of dye administration, a marked erythematous response was observed 24 h after the induction of dermatitis with 5% BZK in rat ears (Figure 2A) which was notably prevented by topical application of DAPS (2.5%) (Figure 2B). A detailed histological characterization of the inflammatory response confirmed that topic application of DAPS (2.5%) reduced the extensive skin edema (Figure 2C-D) and leukocyte infiltration (Figure 2E-H) associated with dermatitis in untreated animals. Dermatomyositis was also observed in the erector muscle of the ear after BZK-induced dermatitis (Figure 2I) and likewise, it was prevented by DAPS administration (Figure 2J). The involvement of leukocyte infiltration in this inflammatory response was confirmed by the significant elevation of MPO activity. This increase was prevented by topical administration of DHPS (5%), and more effectively with 2.5% DAPS (Figure 2K).Figure 2

Bottom Line: Topical DAPS is more effective than DHPS in preventing inflammatory signs (increased vascular permeability, edema, leukocyte infiltration, MPO activation) caused by contact dermatitis induction in rat ears.DAPS, but not DHPS, effectively inhibits COX-1 and COX-2 activities.DAPS also reduces the increase in serum cytokine concentration induced by lipopolysaccharide in rats.

View Article: PubMed Central - PubMed

Affiliation: Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Hospital Universitario Ramón y Cajal, Madrid, Spain. javier.angulo@hrc.es.

ABSTRACT

Background: Dobesilate (2,5-dihydroxyphenyl sulfonate, DHPS) was recently identified as the most potent member of a family of fibroblast growth factor (FGF) inhibitors headed by gentisic acid, one of the main catabolites of aspirin. Although FGFs were first described as inducers of angiogenesis, they were soon recognized as broad spectrum mitogens. Furthermore, in the last decade these proteins have been shown to participate directly in the onset of inflammation, and their potential angiogenic activity often contributes to the inflammatory process in vivo. The aim of this work was to evaluate the anti-inflammatory, anti-angiogenic and anti-tumoral activities of the derivative of DHPS obtained by acetoxylation of its two hydroxyl groups (2,5-diacetoxyphenyl sulfonate; DAPS).

Methods: Anti-inflammatory, anti-angiogenic and anti-tumoral activities of DHPS and DAPS were compared using in vivo assays of dermatitis, angiogenesis and tumorigenesis. The effects of both compounds on myeloperoxidase (MPO) and cyclooxygenase (COX) activities, cytokine production and FGF-induced fibroblast proliferation were also determined.

Results: Topical DAPS is more effective than DHPS in preventing inflammatory signs (increased vascular permeability, edema, leukocyte infiltration, MPO activation) caused by contact dermatitis induction in rat ears. DAPS, but not DHPS, effectively inhibits COX-1 and COX-2 activities. DAPS also reduces the increase in serum cytokine concentration induced by lipopolysaccharide in rats. Furthermore, DAPS displays higher in vivo efficacy than DHPS in inhibiting FGF-induced angiogenesis and heterotopic glioma progression, with demonstrated oral efficacy to combat both processes.

Conclusions: By inhibiting both FGF-signaling and COX-mediated prostaglandin synthesis, DAPS efficiently breaks the vicious circle created by the reciprocal induction of FGF and prostaglandins, which probably sustains undesirable inflammation in many circumstances. Our findings define the enhancement of anti-inflammatory, anti-angiogenic and anti-tumoral activities by diacetyloxyl derivatization of the FGF inhibitor, dobesilate.

Show MeSH
Related in: MedlinePlus