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Development and Evaluation of a Topical Gel Formulation of 5-Flurouracil  using SuccinylChitosan for the Treatment of Actinic Keratosis V. K. Mourya, N. N. Inamdar, K. R. Shaha, S. S. Kulthe,  V. G. Deshpande* vaibhav_82gd@yahoo.in GOVERNMENT COLLEGE OF PHARMACY, AURANGABAD, (M.S.) INDIA. INTRODUCTION 6. Evaluation of in-vitro release: In-vitro release of  5-FU From gel formulation was studied on Keshary-Chein diffusion cell using cellophane membrane. Chitosan [poly (1->4)-2-amino-2-deoxy-β-D-glucan] is a cationic polysaccharide made by alkaline N-deacetylation of chitin and shows good biocompatibility and biodegradability. Major limitation with the use of chitosan is its poor water-solubility. Hence, water-soluble derivatives of chitosan are garnering attention recently. Also, chitosan derivatives have demonstrated unique biological activities including antitumor, antiulcer, immuno-stimulatory, and antibacterial. One such promising candidate is succinylchitosan (Suc-Chi). Actinic keratosis (AK) is a UV light-induced lesion of the skin that may progress to invasive squamous cell carcinoma. 5-Flurouracil (5-FU) is one of the most frequently used drugs for the treatment of AK as it causes cell death in actively proliferating cells by inhibiting thymidylatesynthetase. Several formulations of 5-FU are available, including a 5% cream or solution, a 2% solution, a 1% cream or solution, and, most recently, a micronized 0.5% cream. RESULTS AND DISCUSSION: Objectives Combine anticancer and penetration enhancing activities of Suc-Chi with 5-FU activity and reduce the dose of drug required during formulation. To evaluate the formulated gel against AK using SK-MEL-2 cell line and ascertain the cytotoxic activity of prepared 5-FU topical gel against the same. Formation of Suc-Chi was confirmed by IR in the range of 410-425nm.  CELL TOXICITY STUDY OF SUCCINYL CHITOSAN AND 5-FU EXPERIMENTAL The synergistic effect on cell toxicity seen foresees that such kind of gel formulation for the topical application of cytotoxic drug can be develop successfully.  Synthesis and optimization of synthetic route for Suc-Chi from chitosan. Determination of physicochemical parameters of Suc-Chi. Cell toxicity study of Suc-Chi and 5-FU. Development of Suc-Chi based topical drug delivery. Performance evaluation of the developed product. 1. Synthesis of Suc-Chi:  Chitosan in aqueous acetic acid  was reacted with succinic anhydride for 24 hrs at  400C. Reaction mixture was cooled to room temperature, and precipitated in excess of acetone and dried. 2.Characterization of Suc-Chi: Suc-Chi was characterized by FTIR in the range of 400-4000 cm-1, DSC from 75- 400°C under continuous flow of dry nitrogen gas (50 ml/ min) at  heating rate of 20°C/ min. and by NMR spectroscopy. 3.Cell toxicity study and antitumor activity of Suc-Chi and 5-FU Cell toxicity study and was ascertained on normal fibroblast cell and SK-MEL-2 cell line. 4. Formulation of nanosilver gel: To 10 ml of 0.02% nanosilver solution, 100 mg of carbopol 940 was added to get 1% aqueous carbopol gel loaded with nanosilver. Polyethylene glycol was used as humectants in the concentration of 5% w/v. required viscosity was built by drop wise addition of 10%v/v triethnolamine solution. The resulting gel was sonicated for 30 minutes to assure the uniform distribution of nanosilver through the gel. Evaluation of in-vitro release: from the in vitro drug release study Formulation 7 was found to be having content uniformity 95.00 % with good spreadibility and viscosity 1.533 CONCLUSION The evaluation of formulated gel against AK using specific cell line was the major objective of the present study. Cytotoxic activity of prepared 5-FU topical gel was evaluated against SK-MEL-2 cell line. Our findings suggest that the developed topical gel formulation of 5-FU with Suc-Chi demonstrates synergistic cytotoxic effect on the cancerous cells with a reduction in dose. Also the study can be extended to other types of cell lines. ACKNOWLEDGEMENT The authors are thankful to ICPA  and K-LAB for their generous support REFERENCES Aiping Z., Tian C., Lanhua Y., Hao W., Ping L. Synthesis and characterization of N-succinyl-chitosan and its self-assembly of nanospheresCarbohydr. Polym.2006, 66, 274-279. International Conference on Recent Advances in Cancer Research: Bench to Bedside February 19-20, 2011
Development and Evaluation of a Topical Gel Formulation of 5-Flurouracil  using SuccinylChitosan for the Treatment of Actinic Keratosis V. K. Mourya, N. N. Inamdar, K. R. Shaha, S. S. Kulthe,  V. G. Deshpande* vaibhav_82gd@yahoo.in GOVERNMENT COLLEGE OF PHARMACY, AURANGABAD, (M.S.) INDIA. INTRODUCTION 5. Evaluation of  the developed formulations: All the developed formulations were evaluated for consistency, clarity, pH, spreadability, viscosity, drug content, and in vitro drug release (Keshary-Chein diffusion cell using cellophane membrane). Chitosan [poly (1->4)-2-amino-2-deoxy-β-D-glucan] is a cationic polysaccharide made by alkaline N-deacetylation of chitin and shows good biocompatibility and biodegradability. Major limitation with the use of chitosan is its poor water-solubility. Hence, water-soluble derivatives of chitosan are garnering attention recently. Also, chitosan derivatives have demonstrated unique biological activities including antitumor, antiulcer, immuno-stimulatory, and antibacterial. One such promising candidate is succinylchitosan (Suc-Chi). Actinic keratosis (AK) is a UV light-induced lesion of the skin that may progress to invasive squamous cell carcinoma. 5-Flurouracil (5-FU) is one of the most frequently used drugs for the treatment of AK as it causes cell death in actively proliferating cells by inhibiting thymidylatesynthetase. Several formulations of 5-FU are available, including a 5% cream or solution, a 2% solution, a 1% cream or solution, and, most recently, a micronized 0.5% cream. RESULTS AND DISCUSSION: Formation of Suc-Chi was confirmed by IR in the range of 410-425nm,  Objectives Combine anticancer and penetration enhancing activities of Suc-Chi with 5-FU activity and reduce the dose of drug required during formulation. To evaluate the formulated gel against AK using SK-MEL-2 cell line and ascertain the cytotoxic activity of prepared 5-FU topical gel against the same. EXPERIMENTAL Synthesis and optimization of synthetic route for Suc-Chi from chitosan. Determination of physicochemical parameters of Suc-Chi. Cell toxicity study of Suc-Chi and 5-FU. Development of Suc-Chi based topical drug delivery. Performance evaluation of the developed product. 1. Synthesis of Suc-Chi:  Chitosan in aqueous acetic acid  was reacted with succinic anhydride for 24 hrs at  400C. Reaction mixture was cooled to room temperature, and precipitated in excess of acetone and dried. 2.Characterization of Suc-Chi: Suc-Chi was characterized by FTIR in the range of 400-4000 cm-1, DSC from 75- 400°C under continuous flow of dry nitrogen gas (50 ml/ min) at  heating rate of 20°C/ min. and by NMR spectroscopy. 3.Cell toxicity study and antitumor activity of Suc-Chi and 5-FU: Cell toxicity study was performed on normal fibroblast cell and SK-MEL-2 cell line. 4. Formulation of nanosilver gel: To 10 ml of 0.02% nanosilver solution, 100 mg of carbopol 940 was added to get 1% aqueous carbopol gel loaded with nanosilver. Polyethylene glycol was used as humectant at a concentration of 5% w/v. Viscosity was built by drop wise addition of 10%v/v triethanolamine solution. The resulting gel was sonicated for 30 minutes to assure uniform distribution of nanosilver throughout the gel. CELL TOXICITY STUDY OF SUCCINYL CHITOSAN AND 5-FU The synergistic effect on cell toxicity seen foresees that such kind of gel formulation for the topical application of cytotoxic drug can be develop successfully.  Evaluation of  the developed formulations: From the evaluation of all the developed formulations Batch F7 was found to be smooth, transparent with good spreadibility, pH 7.04, viscosity 1.533 mPa and having content uniformity 95.00 %.  CONCLUSION The synergistic effect on cell toxicity seen foresees that such kind of gel formulation for the topical application of cytotoxic drug can be develop successfully.  ACKNOWLEDGEMENT The authors are thankful to ICPA  and K-LAB for their generous support REFERENCES Aiping Z., Tian C., Lanhua Y., Hao W., Ping L., Synthesis and characterization of N-succinyl-chitosan and its self-assembly of nanospheresCarbohydr. Polym.2006, 66, 274-279.

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Succ chi poster

  • 1. Development and Evaluation of a Topical Gel Formulation of 5-Flurouracil using SuccinylChitosan for the Treatment of Actinic Keratosis V. K. Mourya, N. N. Inamdar, K. R. Shaha, S. S. Kulthe, V. G. Deshpande* vaibhav_82gd@yahoo.in GOVERNMENT COLLEGE OF PHARMACY, AURANGABAD, (M.S.) INDIA. INTRODUCTION 6. Evaluation of in-vitro release: In-vitro release of 5-FU From gel formulation was studied on Keshary-Chein diffusion cell using cellophane membrane. Chitosan [poly (1->4)-2-amino-2-deoxy-β-D-glucan] is a cationic polysaccharide made by alkaline N-deacetylation of chitin and shows good biocompatibility and biodegradability. Major limitation with the use of chitosan is its poor water-solubility. Hence, water-soluble derivatives of chitosan are garnering attention recently. Also, chitosan derivatives have demonstrated unique biological activities including antitumor, antiulcer, immuno-stimulatory, and antibacterial. One such promising candidate is succinylchitosan (Suc-Chi). Actinic keratosis (AK) is a UV light-induced lesion of the skin that may progress to invasive squamous cell carcinoma. 5-Flurouracil (5-FU) is one of the most frequently used drugs for the treatment of AK as it causes cell death in actively proliferating cells by inhibiting thymidylatesynthetase. Several formulations of 5-FU are available, including a 5% cream or solution, a 2% solution, a 1% cream or solution, and, most recently, a micronized 0.5% cream. RESULTS AND DISCUSSION: Objectives Combine anticancer and penetration enhancing activities of Suc-Chi with 5-FU activity and reduce the dose of drug required during formulation. To evaluate the formulated gel against AK using SK-MEL-2 cell line and ascertain the cytotoxic activity of prepared 5-FU topical gel against the same. Formation of Suc-Chi was confirmed by IR in the range of 410-425nm. CELL TOXICITY STUDY OF SUCCINYL CHITOSAN AND 5-FU EXPERIMENTAL The synergistic effect on cell toxicity seen foresees that such kind of gel formulation for the topical application of cytotoxic drug can be develop successfully. Synthesis and optimization of synthetic route for Suc-Chi from chitosan. Determination of physicochemical parameters of Suc-Chi. Cell toxicity study of Suc-Chi and 5-FU. Development of Suc-Chi based topical drug delivery. Performance evaluation of the developed product. 1. Synthesis of Suc-Chi: Chitosan in aqueous acetic acid was reacted with succinic anhydride for 24 hrs at 400C. Reaction mixture was cooled to room temperature, and precipitated in excess of acetone and dried. 2.Characterization of Suc-Chi: Suc-Chi was characterized by FTIR in the range of 400-4000 cm-1, DSC from 75- 400°C under continuous flow of dry nitrogen gas (50 ml/ min) at heating rate of 20°C/ min. and by NMR spectroscopy. 3.Cell toxicity study and antitumor activity of Suc-Chi and 5-FU Cell toxicity study and was ascertained on normal fibroblast cell and SK-MEL-2 cell line. 4. Formulation of nanosilver gel: To 10 ml of 0.02% nanosilver solution, 100 mg of carbopol 940 was added to get 1% aqueous carbopol gel loaded with nanosilver. Polyethylene glycol was used as humectants in the concentration of 5% w/v. required viscosity was built by drop wise addition of 10%v/v triethnolamine solution. The resulting gel was sonicated for 30 minutes to assure the uniform distribution of nanosilver through the gel. Evaluation of in-vitro release: from the in vitro drug release study Formulation 7 was found to be having content uniformity 95.00 % with good spreadibility and viscosity 1.533 CONCLUSION The evaluation of formulated gel against AK using specific cell line was the major objective of the present study. Cytotoxic activity of prepared 5-FU topical gel was evaluated against SK-MEL-2 cell line. Our findings suggest that the developed topical gel formulation of 5-FU with Suc-Chi demonstrates synergistic cytotoxic effect on the cancerous cells with a reduction in dose. Also the study can be extended to other types of cell lines. ACKNOWLEDGEMENT The authors are thankful to ICPA and K-LAB for their generous support REFERENCES Aiping Z., Tian C., Lanhua Y., Hao W., Ping L. Synthesis and characterization of N-succinyl-chitosan and its self-assembly of nanospheresCarbohydr. Polym.2006, 66, 274-279. International Conference on Recent Advances in Cancer Research: Bench to Bedside February 19-20, 2011
  • 2. Development and Evaluation of a Topical Gel Formulation of 5-Flurouracil using SuccinylChitosan for the Treatment of Actinic Keratosis V. K. Mourya, N. N. Inamdar, K. R. Shaha, S. S. Kulthe, V. G. Deshpande* vaibhav_82gd@yahoo.in GOVERNMENT COLLEGE OF PHARMACY, AURANGABAD, (M.S.) INDIA. INTRODUCTION 5. Evaluation of the developed formulations: All the developed formulations were evaluated for consistency, clarity, pH, spreadability, viscosity, drug content, and in vitro drug release (Keshary-Chein diffusion cell using cellophane membrane). Chitosan [poly (1->4)-2-amino-2-deoxy-β-D-glucan] is a cationic polysaccharide made by alkaline N-deacetylation of chitin and shows good biocompatibility and biodegradability. Major limitation with the use of chitosan is its poor water-solubility. Hence, water-soluble derivatives of chitosan are garnering attention recently. Also, chitosan derivatives have demonstrated unique biological activities including antitumor, antiulcer, immuno-stimulatory, and antibacterial. One such promising candidate is succinylchitosan (Suc-Chi). Actinic keratosis (AK) is a UV light-induced lesion of the skin that may progress to invasive squamous cell carcinoma. 5-Flurouracil (5-FU) is one of the most frequently used drugs for the treatment of AK as it causes cell death in actively proliferating cells by inhibiting thymidylatesynthetase. Several formulations of 5-FU are available, including a 5% cream or solution, a 2% solution, a 1% cream or solution, and, most recently, a micronized 0.5% cream. RESULTS AND DISCUSSION: Formation of Suc-Chi was confirmed by IR in the range of 410-425nm, Objectives Combine anticancer and penetration enhancing activities of Suc-Chi with 5-FU activity and reduce the dose of drug required during formulation. To evaluate the formulated gel against AK using SK-MEL-2 cell line and ascertain the cytotoxic activity of prepared 5-FU topical gel against the same. EXPERIMENTAL Synthesis and optimization of synthetic route for Suc-Chi from chitosan. Determination of physicochemical parameters of Suc-Chi. Cell toxicity study of Suc-Chi and 5-FU. Development of Suc-Chi based topical drug delivery. Performance evaluation of the developed product. 1. Synthesis of Suc-Chi: Chitosan in aqueous acetic acid was reacted with succinic anhydride for 24 hrs at 400C. Reaction mixture was cooled to room temperature, and precipitated in excess of acetone and dried. 2.Characterization of Suc-Chi: Suc-Chi was characterized by FTIR in the range of 400-4000 cm-1, DSC from 75- 400°C under continuous flow of dry nitrogen gas (50 ml/ min) at heating rate of 20°C/ min. and by NMR spectroscopy. 3.Cell toxicity study and antitumor activity of Suc-Chi and 5-FU: Cell toxicity study was performed on normal fibroblast cell and SK-MEL-2 cell line. 4. Formulation of nanosilver gel: To 10 ml of 0.02% nanosilver solution, 100 mg of carbopol 940 was added to get 1% aqueous carbopol gel loaded with nanosilver. Polyethylene glycol was used as humectant at a concentration of 5% w/v. Viscosity was built by drop wise addition of 10%v/v triethanolamine solution. The resulting gel was sonicated for 30 minutes to assure uniform distribution of nanosilver throughout the gel. CELL TOXICITY STUDY OF SUCCINYL CHITOSAN AND 5-FU The synergistic effect on cell toxicity seen foresees that such kind of gel formulation for the topical application of cytotoxic drug can be develop successfully. Evaluation of the developed formulations: From the evaluation of all the developed formulations Batch F7 was found to be smooth, transparent with good spreadibility, pH 7.04, viscosity 1.533 mPa and having content uniformity 95.00 %. CONCLUSION The synergistic effect on cell toxicity seen foresees that such kind of gel formulation for the topical application of cytotoxic drug can be develop successfully. ACKNOWLEDGEMENT The authors are thankful to ICPA and K-LAB for their generous support REFERENCES Aiping Z., Tian C., Lanhua Y., Hao W., Ping L., Synthesis and characterization of N-succinyl-chitosan and its self-assembly of nanospheresCarbohydr. Polym.2006, 66, 274-279.