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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

Effects of intraperitoneal administration of potassium 2,5-dihydroxyphenyl sulfonate (DHPS) and potassium 2,5-diacetoxyphenyl sulfonate (DAPS) on the progression of tumors established in rats by subcutaneous implantation of rat glioma C6 cells (5 × 105cells). Photographs show tumors removed from rats treated with vehicle (0.9% NaCl; n = 11; A), DHPS (100 mg/kg/d; eq. to 0.44 mmol/kg/d, i.p. for 10 days; n = 12; B), or DAPS (100 mg/kg/d, eq. to 0.32 mmol/kg/d, i.p. for 10 days; n = 13; C). Treatment began once the presence of a tumor was verified on the fifth day after glioma cell implantation. Tumors were removed after 10 days of treatment and the empty spaces in panel C correspond to tumors that had completely regressed after DAPS treatment (N.D., not detected). A 10 mm bar is displayed in each image. Panels D and E show the quantification and comparison of the volumes and weights, respectively, of the tumors developed in rats treated with vehicle (VEH), DHPS or DAPS. The data are expressed as the mean ± SEM. numbers of animals are shown in parenthesis. *p < 0.05; **p < 0.01; ***p < 0.001 vs. VEH; † p < 0.05 vs. DHPS, by one-factor ANOVA followed by Student-Newman-Keuls test. Panel F shows the effects of subcutaneous glioma development and the treatment with DHBS and DAPS on serum concentrations of lactate dehydrogenase (LDH) in rats. The data are expressed as the mean ± SEM of the percentage of serum LDH detected in control, not bearing glioma, rats (CON). Numbers of animals are shown in parenthesis. *p < 0.05; **p < 0.01; vs. CON; † p < 0.05 vs. VEH, by one-factor ANOVA followed by Student-Newman-Keuls test.
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Fig6: Effects of intraperitoneal administration of potassium 2,5-dihydroxyphenyl sulfonate (DHPS) and potassium 2,5-diacetoxyphenyl sulfonate (DAPS) on the progression of tumors established in rats by subcutaneous implantation of rat glioma C6 cells (5 × 105cells). Photographs show tumors removed from rats treated with vehicle (0.9% NaCl; n = 11; A), DHPS (100 mg/kg/d; eq. to 0.44 mmol/kg/d, i.p. for 10 days; n = 12; B), or DAPS (100 mg/kg/d, eq. to 0.32 mmol/kg/d, i.p. for 10 days; n = 13; C). Treatment began once the presence of a tumor was verified on the fifth day after glioma cell implantation. Tumors were removed after 10 days of treatment and the empty spaces in panel C correspond to tumors that had completely regressed after DAPS treatment (N.D., not detected). A 10 mm bar is displayed in each image. Panels D and E show the quantification and comparison of the volumes and weights, respectively, of the tumors developed in rats treated with vehicle (VEH), DHPS or DAPS. The data are expressed as the mean ± SEM. numbers of animals are shown in parenthesis. *p < 0.05; **p < 0.01; ***p < 0.001 vs. VEH; † p < 0.05 vs. DHPS, by one-factor ANOVA followed by Student-Newman-Keuls test. Panel F shows the effects of subcutaneous glioma development and the treatment with DHBS and DAPS on serum concentrations of lactate dehydrogenase (LDH) in rats. The data are expressed as the mean ± SEM of the percentage of serum LDH detected in control, not bearing glioma, rats (CON). Numbers of animals are shown in parenthesis. *p < 0.05; **p < 0.01; vs. CON; † p < 0.05 vs. VEH, by one-factor ANOVA followed by Student-Newman-Keuls test.

Mentions: Considering the above confirmed capacity of DHPS and DAPS to inhibit angiogenesis and inflammation and given the key roles of these processes in carcinogenesis, the effects of DHPS and DAPS on tumoral progression were evaluated. To estimate the relative anti-tumoral potential of these two compounds, we used a well-known heterotopic model of glioma. As seen in Figure 6A-C, gliomas in rats treated for 10 days with vehicle (0.9% NaCl) were larger than those in rats treated with DHPS (100 mg/kg/day; i.p.) or DAPS (100 mg/kg/day; i.p.). Rats that received DHPS also exhibited larger tumors than those treated with DAPS, and indeed, four rats that received DAPS were tumor-free by the end of treatment, despite the fact that all the rats developed comparable tumors prior to treatment. By contrast, none of the tumors were fully eliminated in the animals administered the vehicle alone or DHPS. Quantification of the tumor volume confirmed that tumor progression was significantly inhibited in DHPS-treated rats, an effect that was further enhanced in DAPS-treated rats (Figure 6D). Evaluation of the tumor weight yielded similar results, although the reduction in weight was only significant in rats treated with DAPS (Figure 6E). This attenuated effect may be explained by the existence of necrotic processes in vehicle-treated tumors, a frequently-observed phenomenon that diminishes tumor density as the tumor volume increases. In fact, the serum concentrations of lactate dehydrogenase (LDH), which is a marker of tumoral necrosis [35,36], are significantly increased in vehicle treated rats with subcutaneous gliomas when compared to healthy not bearing glioma rats. Serum LDH levels in glioma implanted rats were significantly reduced by DHPS treatment and normalized by treating with DAPS (Figure 6F).Figure 6


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)

Effects of intraperitoneal administration of potassium 2,5-dihydroxyphenyl sulfonate (DHPS) and potassium 2,5-diacetoxyphenyl sulfonate (DAPS) on the progression of tumors established in rats by subcutaneous implantation of rat glioma C6 cells (5 × 105cells). Photographs show tumors removed from rats treated with vehicle (0.9% NaCl; n = 11; A), DHPS (100 mg/kg/d; eq. to 0.44 mmol/kg/d, i.p. for 10 days; n = 12; B), or DAPS (100 mg/kg/d, eq. to 0.32 mmol/kg/d, i.p. for 10 days; n = 13; C). Treatment began once the presence of a tumor was verified on the fifth day after glioma cell implantation. Tumors were removed after 10 days of treatment and the empty spaces in panel C correspond to tumors that had completely regressed after DAPS treatment (N.D., not detected). A 10 mm bar is displayed in each image. Panels D and E show the quantification and comparison of the volumes and weights, respectively, of the tumors developed in rats treated with vehicle (VEH), DHPS or DAPS. The data are expressed as the mean ± SEM. numbers of animals are shown in parenthesis. *p < 0.05; **p < 0.01; ***p < 0.001 vs. VEH; † p < 0.05 vs. DHPS, by one-factor ANOVA followed by Student-Newman-Keuls test. Panel F shows the effects of subcutaneous glioma development and the treatment with DHBS and DAPS on serum concentrations of lactate dehydrogenase (LDH) in rats. The data are expressed as the mean ± SEM of the percentage of serum LDH detected in control, not bearing glioma, rats (CON). Numbers of animals are shown in parenthesis. *p < 0.05; **p < 0.01; vs. CON; † p < 0.05 vs. VEH, by one-factor ANOVA followed by Student-Newman-Keuls test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4318172&req=5

Fig6: Effects of intraperitoneal administration of potassium 2,5-dihydroxyphenyl sulfonate (DHPS) and potassium 2,5-diacetoxyphenyl sulfonate (DAPS) on the progression of tumors established in rats by subcutaneous implantation of rat glioma C6 cells (5 × 105cells). Photographs show tumors removed from rats treated with vehicle (0.9% NaCl; n = 11; A), DHPS (100 mg/kg/d; eq. to 0.44 mmol/kg/d, i.p. for 10 days; n = 12; B), or DAPS (100 mg/kg/d, eq. to 0.32 mmol/kg/d, i.p. for 10 days; n = 13; C). Treatment began once the presence of a tumor was verified on the fifth day after glioma cell implantation. Tumors were removed after 10 days of treatment and the empty spaces in panel C correspond to tumors that had completely regressed after DAPS treatment (N.D., not detected). A 10 mm bar is displayed in each image. Panels D and E show the quantification and comparison of the volumes and weights, respectively, of the tumors developed in rats treated with vehicle (VEH), DHPS or DAPS. The data are expressed as the mean ± SEM. numbers of animals are shown in parenthesis. *p < 0.05; **p < 0.01; ***p < 0.001 vs. VEH; † p < 0.05 vs. DHPS, by one-factor ANOVA followed by Student-Newman-Keuls test. Panel F shows the effects of subcutaneous glioma development and the treatment with DHBS and DAPS on serum concentrations of lactate dehydrogenase (LDH) in rats. The data are expressed as the mean ± SEM of the percentage of serum LDH detected in control, not bearing glioma, rats (CON). Numbers of animals are shown in parenthesis. *p < 0.05; **p < 0.01; vs. CON; † p < 0.05 vs. VEH, by one-factor ANOVA followed by Student-Newman-Keuls test.
Mentions: Considering the above confirmed capacity of DHPS and DAPS to inhibit angiogenesis and inflammation and given the key roles of these processes in carcinogenesis, the effects of DHPS and DAPS on tumoral progression were evaluated. To estimate the relative anti-tumoral potential of these two compounds, we used a well-known heterotopic model of glioma. As seen in Figure 6A-C, gliomas in rats treated for 10 days with vehicle (0.9% NaCl) were larger than those in rats treated with DHPS (100 mg/kg/day; i.p.) or DAPS (100 mg/kg/day; i.p.). Rats that received DHPS also exhibited larger tumors than those treated with DAPS, and indeed, four rats that received DAPS were tumor-free by the end of treatment, despite the fact that all the rats developed comparable tumors prior to treatment. By contrast, none of the tumors were fully eliminated in the animals administered the vehicle alone or DHPS. Quantification of the tumor volume confirmed that tumor progression was significantly inhibited in DHPS-treated rats, an effect that was further enhanced in DAPS-treated rats (Figure 6D). Evaluation of the tumor weight yielded similar results, although the reduction in weight was only significant in rats treated with DAPS (Figure 6E). This attenuated effect may be explained by the existence of necrotic processes in vehicle-treated tumors, a frequently-observed phenomenon that diminishes tumor density as the tumor volume increases. In fact, the serum concentrations of lactate dehydrogenase (LDH), which is a marker of tumoral necrosis [35,36], are significantly increased in vehicle treated rats with subcutaneous gliomas when compared to healthy not bearing glioma rats. Serum LDH levels in glioma implanted rats were significantly reduced by DHPS treatment and normalized by treating with DAPS (Figure 6F).Figure 6

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