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Antiproliferative property of hexadecyloxypropyl 9-[2-(phosphonomethoxy) ethyl] guanine (HDP-PMEG) for unwanted ocular proliferation.

Hou J, Li Y, Zhou Z, Valiaeva N, Beadle JR, Hostetler K, Freeman WR, Hu DN, Chen H, Cheng L - Mol. Vis. (2011)

Bottom Line: Its inhibitory effects on those cells were uniformly stronger than that of 5-FU.The safety profile of HDP-PMEG was significantly better than 5-FU and daunorubicin.HDP-PMEG possesses a significant inhibitory effect on the proliferation of RPE, retinal glial cells, scleral fibroblasts, and ocular melanoma cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Ocular Pharmacology, School of Ophthalmology and Optometry, Wenzhou Medical College, Wenzhou, Zhejiang,China.

ABSTRACT

Purpose: To investigate the safety and inhibitory effects of hexadecyloxypropyl 9-[2-(phosphonomethoxy) ethyl] guanine (HDP-PMEG) on ocular cell proliferation and collagen matrix contraction.

Methods: For the antiproliferation studies, various ocular cell monolayers were exposed to HDP-PMEG, PMEG, 5-fluorouracil (5-FU), and daunorubicin (DNB). For the collagen contraction studies, retinal pigment epithelium (RPE) cells seeded onto type I collagen lattices were exposed for a single 5- or 50-min period to various concentrations of HDP-PMEG or 5-FU. For the cytotoxicity study, trypan blue exclusion tests were performed using a human Müller cell line. Cytotoxicity was determined up to 4 days after treatment.

Results: The proliferation of RPE cells, scleral fibroblasts, vessel endothelial cells, and ocular melanoma cells can all be significantly inhibited by HDP-PMEG. Its inhibitory effects on those cells were uniformly stronger than that of 5-FU. Contraction of the collagen matrix containing RPE cells was significantly inhibited by HDP-PMEG and by 5-FU at concentrations of 20 µM and 2,000 µM, respectively, as compared with controls (p<0.05). The safety profile of HDP-PMEG was significantly better than 5-FU and daunorubicin. The ocular therapeutic index is 1,100 for HDP-PMEG, 17.2 for 5-FU, and 1.25 for daunorubicin.

Conclusions: HDP-PMEG possesses a significant inhibitory effect on the proliferation of RPE, retinal glial cells, scleral fibroblasts, and ocular melanoma cells. HDP-PMEG is also genotoxic and may be used as a single short application for the modulation of unwanted ocular proliferation.

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Cytotoxicity assay on Müller cell. A: The percentage of trypan blue positive cells (y-axis) was plotted against the concentration of the test compounds used (x-axis). The concentration unit was micro molar and the open box represented a 95% confidence limit. 5-FU at 100 µM that was the highest concentration tested, caused 20% death of the cell culture, which was equivalent to HDP-PMEG at 11 µM or DNR at 0.1 µM. B: Exemplary images from the Müller cell cytotoxicity study: the upper left panel showing a counting field from the control in which 31 cells were stained by trypan blue out of a total of 435 cells, yielding a 7% trypan blue positive rate. The rest of the three fields had a trypan blue positive rate of 19% (38/198) for 10 µM HDP-PMEG, 18% (46/251) for 100 µM 5-FU, and 18% (49/274) for 0.1 µM daunorubicin (DNR).
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f4: Cytotoxicity assay on Müller cell. A: The percentage of trypan blue positive cells (y-axis) was plotted against the concentration of the test compounds used (x-axis). The concentration unit was micro molar and the open box represented a 95% confidence limit. 5-FU at 100 µM that was the highest concentration tested, caused 20% death of the cell culture, which was equivalent to HDP-PMEG at 11 µM or DNR at 0.1 µM. B: Exemplary images from the Müller cell cytotoxicity study: the upper left panel showing a counting field from the control in which 31 cells were stained by trypan blue out of a total of 435 cells, yielding a 7% trypan blue positive rate. The rest of the three fields had a trypan blue positive rate of 19% (38/198) for 10 µM HDP-PMEG, 18% (46/251) for 100 µM 5-FU, and 18% (49/274) for 0.1 µM daunorubicin (DNR).

Mentions: Figure 4A demonstrates the drug cytotoxicity results from the trypan blue dye exclusion assay. All three drugs showed a higher mean percentage of trypan blue positive cells as compared to the controls even at a 0.001 µM concentration (5-FU: 8.8% versus 7.6%, p=0.0353; DNR: 14.6% versus 7.6%, p<0.0001; HDP-PMEG: 10% versus 7.6%, p=0.0176 Adjusted for multiple comparisons with Dunnett). The TC50 (the concentration to cause 50% cell death) for HDP-PMEG and 5-FU was not reached in this study and for DNR was 3 µM; however, the TC20 was available from the study and it was 11 µM for HDP-PMEG, 100 µM for 5-FU, and 0.1 µM for DNR (Figure 4B). To estimate the ocular therapeutic index (TI=TC50/IC50) which is defined as the ratio of drug efficacy to the magnitude of its adverse side effects, we used TC20 and IC20 to calculate the TI. TI was 17.2 for 5-FU, 1,100 for HDP-PMEG, and 1.25 for DNR. A high ocular TI will suggest that the compound might prevent proliferation of cells in vivo without affecting non-dividing cells such as those of the retina.


Antiproliferative property of hexadecyloxypropyl 9-[2-(phosphonomethoxy) ethyl] guanine (HDP-PMEG) for unwanted ocular proliferation.

Hou J, Li Y, Zhou Z, Valiaeva N, Beadle JR, Hostetler K, Freeman WR, Hu DN, Chen H, Cheng L - Mol. Vis. (2011)

Cytotoxicity assay on Müller cell. A: The percentage of trypan blue positive cells (y-axis) was plotted against the concentration of the test compounds used (x-axis). The concentration unit was micro molar and the open box represented a 95% confidence limit. 5-FU at 100 µM that was the highest concentration tested, caused 20% death of the cell culture, which was equivalent to HDP-PMEG at 11 µM or DNR at 0.1 µM. B: Exemplary images from the Müller cell cytotoxicity study: the upper left panel showing a counting field from the control in which 31 cells were stained by trypan blue out of a total of 435 cells, yielding a 7% trypan blue positive rate. The rest of the three fields had a trypan blue positive rate of 19% (38/198) for 10 µM HDP-PMEG, 18% (46/251) for 100 µM 5-FU, and 18% (49/274) for 0.1 µM daunorubicin (DNR).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3049735&req=5

f4: Cytotoxicity assay on Müller cell. A: The percentage of trypan blue positive cells (y-axis) was plotted against the concentration of the test compounds used (x-axis). The concentration unit was micro molar and the open box represented a 95% confidence limit. 5-FU at 100 µM that was the highest concentration tested, caused 20% death of the cell culture, which was equivalent to HDP-PMEG at 11 µM or DNR at 0.1 µM. B: Exemplary images from the Müller cell cytotoxicity study: the upper left panel showing a counting field from the control in which 31 cells were stained by trypan blue out of a total of 435 cells, yielding a 7% trypan blue positive rate. The rest of the three fields had a trypan blue positive rate of 19% (38/198) for 10 µM HDP-PMEG, 18% (46/251) for 100 µM 5-FU, and 18% (49/274) for 0.1 µM daunorubicin (DNR).
Mentions: Figure 4A demonstrates the drug cytotoxicity results from the trypan blue dye exclusion assay. All three drugs showed a higher mean percentage of trypan blue positive cells as compared to the controls even at a 0.001 µM concentration (5-FU: 8.8% versus 7.6%, p=0.0353; DNR: 14.6% versus 7.6%, p<0.0001; HDP-PMEG: 10% versus 7.6%, p=0.0176 Adjusted for multiple comparisons with Dunnett). The TC50 (the concentration to cause 50% cell death) for HDP-PMEG and 5-FU was not reached in this study and for DNR was 3 µM; however, the TC20 was available from the study and it was 11 µM for HDP-PMEG, 100 µM for 5-FU, and 0.1 µM for DNR (Figure 4B). To estimate the ocular therapeutic index (TI=TC50/IC50) which is defined as the ratio of drug efficacy to the magnitude of its adverse side effects, we used TC20 and IC20 to calculate the TI. TI was 17.2 for 5-FU, 1,100 for HDP-PMEG, and 1.25 for DNR. A high ocular TI will suggest that the compound might prevent proliferation of cells in vivo without affecting non-dividing cells such as those of the retina.

Bottom Line: Its inhibitory effects on those cells were uniformly stronger than that of 5-FU.The safety profile of HDP-PMEG was significantly better than 5-FU and daunorubicin.HDP-PMEG possesses a significant inhibitory effect on the proliferation of RPE, retinal glial cells, scleral fibroblasts, and ocular melanoma cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Ocular Pharmacology, School of Ophthalmology and Optometry, Wenzhou Medical College, Wenzhou, Zhejiang,China.

ABSTRACT

Purpose: To investigate the safety and inhibitory effects of hexadecyloxypropyl 9-[2-(phosphonomethoxy) ethyl] guanine (HDP-PMEG) on ocular cell proliferation and collagen matrix contraction.

Methods: For the antiproliferation studies, various ocular cell monolayers were exposed to HDP-PMEG, PMEG, 5-fluorouracil (5-FU), and daunorubicin (DNB). For the collagen contraction studies, retinal pigment epithelium (RPE) cells seeded onto type I collagen lattices were exposed for a single 5- or 50-min period to various concentrations of HDP-PMEG or 5-FU. For the cytotoxicity study, trypan blue exclusion tests were performed using a human Müller cell line. Cytotoxicity was determined up to 4 days after treatment.

Results: The proliferation of RPE cells, scleral fibroblasts, vessel endothelial cells, and ocular melanoma cells can all be significantly inhibited by HDP-PMEG. Its inhibitory effects on those cells were uniformly stronger than that of 5-FU. Contraction of the collagen matrix containing RPE cells was significantly inhibited by HDP-PMEG and by 5-FU at concentrations of 20 µM and 2,000 µM, respectively, as compared with controls (p<0.05). The safety profile of HDP-PMEG was significantly better than 5-FU and daunorubicin. The ocular therapeutic index is 1,100 for HDP-PMEG, 17.2 for 5-FU, and 1.25 for daunorubicin.

Conclusions: HDP-PMEG possesses a significant inhibitory effect on the proliferation of RPE, retinal glial cells, scleral fibroblasts, and ocular melanoma cells. HDP-PMEG is also genotoxic and may be used as a single short application for the modulation of unwanted ocular proliferation.

Show MeSH
Related in: MedlinePlus