Topical Micro-Emulsion of 5-Fluorouracil by a Twin Screw Processor-Based Novel Continuous Manufacturing Process for the Treatment of Skin Cancer: Preparation and In Vitro and In Vivo Evaluations
Abstract
:1. Introduction
2. Methodology
2.1. Materials
2.2. Methods
2.2.1. Preparation of 5-FU@ME Using TSP
2.2.2. Droplet Size, Polydispersity Index and Zeta Potential
2.2.3. Thermodynamic Stability Studies
2.2.4. Fourier Transform Infrared Spectroscopy (FTIR)
2.2.5. Transmission Electron Microscopy (TEM)
2.2.6. Rheological and pH Measurements
2.2.7. Estimation of 5-FU in 5-FU@MEs Using HPLC
2.2.8. Determination of Drug Content
2.2.9. In Vitro Drug Release Study
2.2.10. Ex Vivo Skin Permeation Studies
2.2.11. In Vitro Cytotoxicity Studies
2.2.12. Skin Irritation and Histopathological Studies
- Group I: Treated with 0.5 g of Blank formulation (no drug)
- Group II: Treated with 0.5 g of Optimized formulation (5-FU@ME-2)
- Group III: Treated with Formalin (0.8% v/v)
2.2.13. In Vivo Pharmacodynamics Studies
- Group 1: Normal Control (no disease induced, no drug treatment)
- Group 2: Positive control (disease induced, no treatment)
- Group 3: Marketed formulation (Flonida 1% w/w)
- Group 4: Optimised Formulation (5-FU@ME-2)
3. Results and Discussion
3.1. Preparation of MEs Using TSP
3.2. Polydispersity Index and Droplet Size
3.3. Thermodynamic Stability Studies
3.4. Fourier Transform Infrared Spectroscopy
3.5. Transmission Electron Microscopy
3.6. Rheology, pH and Drug Content
3.7. In Vitro Drug Release Study
3.8. Ex Vivo Skin Permeation Studies
3.9. In Vitro Cytotoxicity Studies
3.10. Skin Irritation Studies
3.11. In Vivo Pharmacodynamic Studies
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Batch Code | Percent Quantity of Each Component | |||||
---|---|---|---|---|---|---|
Inner Phase | Outer Phase | Total | ||||
5-FU | Water | AOT | IPM | Tween 80 | ||
5-FU@ME-1 | 0.50 | 25 | 18.5 | 50 | 6 | 100.00 |
5-FU@ME-2 | 0.50 | 20 | 18.5 | 55 | 6 | 100.00 |
5-FU@ME-3 | 0.50 | 15 | 18.5 | 60 | 6 | 100.00 |
5-FU@ME-4 | 0.50 | 10 | 18.5 | 65 | 6 | 100.00 |
RPM in TSP | Droplet Size (nm) | PDI | Zeta Potential (mV) |
---|---|---|---|
100 RPM | 178.7 ± 3.22 | 0.247 ± 0.03 | −1.011 ± 0.21 |
150 RPM | 978.1 ± 5.56 | 1.000 ± 0.00 | −18.05 ± 1.09 |
200 RPM | 1601.6 ± 8.76 | 0.977 ± 0.08 | −4.65 ± 0.34 |
250 RPM | 2795.6 ± 9.65 | 1.000 ± 0.00 | −8.54 ± 0.92 |
Systems | Flux (µg/cm2/h) | Q24 (µg) | Drug Content in Skin (µg/cm2) |
---|---|---|---|
5-FU solution | 5.63 ± 0.11 | 132.93 ± 5.11 | 58.32 ± 0.09 |
5-FU@ME2 | 19.63 ± 0.12 | 940.93 ± 5.53 | 294.46 ± 1.12 |
Sr. No. | Sample Code | IC50 (µg/mL) | |
---|---|---|---|
HaCat | A431 | ||
1 | Blank ME | 4.5 | 1.59 |
2 | Pure Drug | 1.3 | 1.08 |
3 | 5-FU@ME-2 | 2.42 | 0.79 |
Formulations | Reaction Grade Observed | Primary Irritation Index | |
---|---|---|---|
Erythema | Oedema | ||
Positive Control | 2.33 ± 0.33 | 2.17 ± 0.31 | 2.25 ± 0.25 |
Blank ME | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 |
5-FU@ME-2 | 0.10 ± 0.10 | 0.09 ± 0.05 | 0.07 ± 0.06 |
Treatment Groups | Degree of Keratinisation | Nuclear Pleomorphism | Mitosis | Inflammatory Infiltration |
---|---|---|---|---|
Group 1 | − | − | − | − |
Group 2 | +++ | ++++ | +++ | +++ |
Group 3 | + | ++ | ++ | +++ |
Group 4 | − | − | − | ++ |
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Nikam, A.N.; Jacob, A.; Raychaudhuri, R.; Fernandes, G.; Pandey, A.; Rao, V.; Ahmad, S.F.; Pannala, A.S.; Mutalik, S. Topical Micro-Emulsion of 5-Fluorouracil by a Twin Screw Processor-Based Novel Continuous Manufacturing Process for the Treatment of Skin Cancer: Preparation and In Vitro and In Vivo Evaluations. Pharmaceutics 2023, 15, 2175. https://doi.org/10.3390/pharmaceutics15092175
Nikam AN, Jacob A, Raychaudhuri R, Fernandes G, Pandey A, Rao V, Ahmad SF, Pannala AS, Mutalik S. Topical Micro-Emulsion of 5-Fluorouracil by a Twin Screw Processor-Based Novel Continuous Manufacturing Process for the Treatment of Skin Cancer: Preparation and In Vitro and In Vivo Evaluations. Pharmaceutics. 2023; 15(9):2175. https://doi.org/10.3390/pharmaceutics15092175
Chicago/Turabian StyleNikam, Ajinkya Nitin, Angela Jacob, Ruchira Raychaudhuri, Gasper Fernandes, Abhijeet Pandey, Vinay Rao, Sheikh F. Ahmad, Ananth S. Pannala, and Srinivas Mutalik. 2023. "Topical Micro-Emulsion of 5-Fluorouracil by a Twin Screw Processor-Based Novel Continuous Manufacturing Process for the Treatment of Skin Cancer: Preparation and In Vitro and In Vivo Evaluations" Pharmaceutics 15, no. 9: 2175. https://doi.org/10.3390/pharmaceutics15092175