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LIPOSOME: AMELIORATIVE TOOL IN TREATMENT OF HYPERPIGMENTATION
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LIPOSOME: AMELIORATIVE TOOL IN TREATMENT OF HYPERPIGMENTATION

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liposomal drug delivery system have great potential to deliver drug at target site

liposomal drug delivery system have great potential to deliver drug at target site

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  • 1. Liposomal drug delivery system Ameliorative tool for treatment of Skin pigmentation Presented by Pournima Mahesh Satpute M.Pharm first semester- 2013 Roll no.-517 Guided by Mrs. Shilpa Shotriya 1
  • 2. Content 1. Introduction to cosmeceutics 2. Pigmentation 3. Factor stimulating melanin synthesis 4. Clinical condition 5. Melanin synthesis 6. Targets 7. Present dosage form 10. Drawback of present dosage form 11.Vesicular drug delivery system –Liposomes 12.Liposomes –Introduction 13.Method of preparation 14.Case studies 15.Conclusion 2
  • 3. Cosmeceuticals word popularized by Albert M. Kligman in the late 1970s  shows beneficial topical actions and provides protection against degenerative skin conditions. Cosmeceuticals is combination of cosmeceutics and pharmaceutics with biologically active ingredients purporting to have medical or drug like benefits. Singhal M., Khanna S, Nasa A.,” COSMECEUTICALS FOR THE SKIN: AN OVERVIEW” Asian Journal of Pharmaceutical and Clinical Research, 2, (2011)1-6 3
  • 4. TYPES OF COSMECEUTICALS COSMECEUTICALS SKIN CARE HAIR COSMECEUTICALS SKIN DEODERISING& PREFUMES Toiletries & soap Anti wrinkle product Moisturizing product Sunscreen product Singhal M., Khanna S, Nasa A.,” COSMECEUTICALS FOR THE SKIN: AN OVERVIEW” Asian Journal of Pharmaceutical and Clinical Research, 4,(2011),1-6 4 Skin whitening product
  • 5. History a reveals that use of different herbal ingredient as skin lightening agent Popularity of skin whitening agent increasing as people getting more conscious about their appearance … Change in Colour of skin frequently raise for medical or cosmetic reason Shoukat Parvez, Moonkyu Kang, Hwan-Suck Chung, Chongwoon Cho, Moo-Chang Hong, Min-Kyu Shin and Hyunsu Bae,” Survey and Mechanism of Skin Depigmenting and Lightening Agents”, PHYTOTHERAPY RESEARCH (2006) 20, 921–934 5 HISTORY
  • 6. Shoukat P., Moonkyu Kang, Hwan-Suck Chung, Chongwoon Cho, Moo-Chang Hong, Min-Kyu Shin and Hyunsu Bae,” Survey and Mechanism of Skin Depigmenting and Lightening Agents”, PHYTOTHERAPY RESEARCH (2006) 20, 921–934 6 Pigmentation •Human skin contains specialized cells, called melanocytes, which are located at the base of the epidermis. These cells are programmed to produce a brown colour pigment called melanin. •It is defense response of the skin to the damaging and potential skin cancer causing rays of the sun.
  • 7. 7
  • 8. Hormonal Oral contraceptive Corticosteroids Pregnancy Radations UV-B Ionizing radiations Miscellaneous Medicines Stress Nina G. Jablonski and George Chaplin,” Human skin pigmentation, migration and disease susceptibility,” Phil. Trans. R. Soc. B (2012), 367, 785–792 8 FACTOR STIMULATING MELANIN SYNTHESIS
  • 9. Nina G. Jablonski and George Chaplin,” Human skin pigmentation, migration and disease susceptibility,” Phil. Trans. R. Soc. B (2012) 367, 785–792. 9  Dark spot appearing with older age on exposure to sun -Lentigines  Irregular grouping of pigment containing cell or melanocytes –Freckles Hormonal hyper pigmentation -Melasma Inflammation related pigmentation CLINICAL conditions:
  • 10. Shoukat Parvez, Moonkyu Kang, Hwan-Suck Chung, Chongwoon Cho, Moo-Chang Hong, Min-Kyu Shin and Hyunsu Bae,” Survey and Mechanism of Skin Depigmenting and Lightening Agents”, PHYTOTHERAPY RESEARCH (2006) 20, 921–934 . 10 MELANIN SYTHESIS
  • 11. 11 TARGET MELANOSOME MATURATION 1. ANTAGONIST OF a MSH 2. a -BISBOLOl TYROSINAS E INHIBITOR 1. GLYCERHIZA GLABRA 2. PHENYL RESORCINOL DENDRIMER FORMATION 1. a –BISBOLOL 2. THIOACETIC ACID MELANOSOME TRANSFER &UPTAKE 1.JACK FRUIT SEED EXTRACT- LECTINS KERATINOCYETE MATURATION &SKIN DESQUAMATION 1. CARICA PAPYA 2,CENTENELA ASIATICA
  • 12. 12 PRESENT DOSAGE FORMS Hydroqinone Magnesium ascorbyl phosphate Monobenzyl ether of hydoqinone Hydroxianisole Kojic acid Niacinamide Arbutin Azelaic acid All this drug available in cream ,lotion for topical application.
  • 13. Marketed Product Ingredient White Radiance Intensive Whitening Cream- Olay niacinamide Demelan- Glenmark Kojic acid 2% + Glycolic acid 10% + Arbutin 5% Cream Eldoquin Forte/Solaquin Forte Hydroquinone 2% +Oxybenzone3% +Octyl Methoxycinnamate 5% Cream Aziderm Azelaic Acid 10% & 20% cream Melacare Forte Hydroquinone 4% + Tretinoin 0.025% +Mometasone Furoate 0.1% Cream Marketed product 13
  • 14. 14 Drawback of Present formulation FREQUENT APPLICATION SHORTER DURATION OF ACTION SYSTEMIC ABSORPTION Solution: develop novel drug delivery system •To achieve drug release at specific sight •Prolong and controlled drug release •Reduce frequency of application
  • 15. Theresa M. Allen a,⁎, Pieter R. Cullis,” Liposomal drug delivery systems: From concept to clinical applications”, Advanced Drug Delivery Reviews (2013) 65, 36–48. 15 VESICULAR DRUG DELIVRY SYSTEM  These are phospholipids based vehicle composed of bilayer membrane.  This system has ability to encapsulate both lipophiic drugs within their membrane and hydrophilic drug inside or outside the aqueous core .  Membrane of these carriers can be altered and tuned.
  • 16. 16 Types of vesicular systems Increase flexibility due to addition of ethanol It shows facilated fusion and malleability with membrane shows superficial drug delivery Designed . Elastic or ultra deforamble system Non ionic surfactant vesicle made up from polyoxy ethylene alkyl ether, polyoxyethylene alkyl esters or saccharose diesters. Rigid vesicles Increase flexibility due to addition of surfactant Elastic or ultra deformable system Rigid vesicles Theresa M. Allen a,⁎, Pieter R.Cullis,” Liposomal drug delivery systems: From concept to clinical applications”, Advanced Drug Delivery Reviews (2013)65 ,36–48.
  • 17. Sharma A, Sharma U., “Liposomes in drug delivery: progress and limitations”, International Journal of Pharmaceutics (1997) 154 ,123 -140 17 Liposomes •Liposomes - Novel Drug Delivery System. •Liposomes were discovered in early 1960’s by Bangham and colleagues. Liposomes were injected to mice in 1970. •The liposome was adopted as a promising delivery system because its organized structure which could hold drugs, depending on their solubility characteristics, in both the aqueous and lipid phases.
  • 18. Sharma A, Sharma U., “Liposomes in drug delivery: progress and limitations”, International Journal of Pharmaceutics (1997) 147,123 -140 18 ADVANTAGE OF LIPOSOMES Increased efficacy and therapeutic index of drug Increased stability via encapsulation Non-toxic, Flexible, Biocompatible, Completely biodegradable, and Nonimmunogenic For systemic and non-systemic administrations reaction Reduce the toxicity of the encapsulated agent Liposomes help reduce the exposure of sensitive tissues to toxic drugs Flexibility to couple with site-specific ligands to achieve active targeting
  • 19. Sharma A, Sharma U., “Liposomes in drug delivery: progress and limitations”, International Journal of Pharmaceutics (1997)147, 123 -140 19 Low solubility Short half-life Sometimes phospholipid undergoes oxidation and hydrolysis-like reaction Leakage and fusion of encapsulated drug/molecules Production cost is high Site avoidance effect demerits
  • 20. 20 Types of Liposomes • Neutral or negatively charge Phospholipids and cholestrol. Conventional liposomes(CL) • Phospholipids such as PE or DOPE with either CHEMS or OA pH sensitive liposomes • Cationic lipids with Dioleolylphosphatidylethanolamine Cationic liposomes • Neutral, high Tc, cholestrol & 5- 10% of PEG - DSPE or GM1 Long circulatory stealth liposomes (LCL) • CL or LCL with attached monoclonal antibody or recognition sequence Immuno-liposomes Based upon Composition & Applications
  • 21. 21 Types of Liposomes 1. MLV Multilamellar large vesicles->0.5m 2. OLV Oligolamellar vesicles-0.1-1 m 3. UV Unilamellar vesicles (all size ranges) 4. SUV Small Unilamellar Vesicles- 20-100nm 5. MUV Medium sized unilamellar vesicles 6. LUV Large unilamellar vesicles->100nm 7. GUV Giant unilamellar vesicles->1m 8. MV Multivesicular vesicles->1m Type Specification
  • 22. Akbarzadeh A., Sadabady R, Davaran S, Joo S., Zarghami S.,Hanifehpour Y., Samiei M., Koshki M., “Liposome: classification, preparation, and applications”, Nanoscale Research Letters 2013, 8,102 22 LIPOSOMES COMPOSITION Liposomes can be formed from a variety of phospholipids. The lipid most widely used is phosphatidyl choline, phosphatidyl ethanolamime and phosphatidlyl serine either as such or in combination with other substance to vary liposome's physical, chemical and biological properties, liposome size, charge, drug loading capacity and permeability.
  • 23. Kulkarni P., Yadav J., Vaidya K., “LIPOSOMES: A NOVEL DRUG DELIVERY SYSTEM”, International Journal of Current Pharmaceutical Research, ( 2011)4(2),10-18 23 Types of phospholipids Example 1. Phospholipids from natural source Egg yolk, Soya bean, PC-Phosphatidylcholine PE- Phosphatidylethanolamine PS- Phosphatidylserine 2) Modified natural phospholipids Partially or Fully hydrogenated natural phosphotidylcholins 3) Semisyntheyic phospholipids Acylated natural phospholipids 4) Fully synthetic phospholipids DOPC- Dioleolylphosphatidylcholine DSPC- Distearoylphosphatidylcholine DOPE- Dioleolylphosphatidylethanolamine DSPE- Distearoylphosphatidylethanolamine 5) . Phospholipids with nonatural head groups PEG linked with phosphotidylcholins
  • 24. 24 Method of preparation General methods of preparation All the methods of preparing the liposomes involve four basic stages: 1. Drying down lipids from organic solvent. 2. Dispersing the lipid in aqueous media. 3. Purifying the resultant liposome. 4. Analyzing the final product. Kulkarni P., Yadav J., Vaidya K., “LIPOSOMES: A NOVEL DRUG DELIVERY SYSTEM”, International Journal of Current Pharmaceutical Research, ( 2011)4(2),10-18
  • 25. Kulkarni P., Yadav J., Vaidya K., “LIPOSOMES: A NOVEL DRUG DELIVERY SYSTEM”, International Journal of Current Pharmaceutical Research, ( 2011)4(2),10-18 25
  • 26. Akbarzadeh A., Sadabady R, Davaran S, Joo S., Zarghami S.,Hanifehpour Y., Samiei M., Koshki M., “Liposome: classification, preparation, and applications”, Nanoscale Research Letters 2013, 8:102 26 The correct choice of liposome preparation method depends on the following parameters: 1) The physicochemical characteristics of the material to be entrapped and those of the liposomal ingredients. 2)The nature of the medium in which the lipid vesicles are dispersed. 3) The effective concentration of the entrapped substance and its potential toxicity . 4)Additional processes involved during application/delivery of the vesicles. 5) Optimum size, polydispersity and shelf-life of the vesicles for the intended application. 6) Batch-to-batch reproducibility and possibility of large-scale production of safe and efficient liposomal product .
  • 27. Akbarzadeh A., Sadabady R, Davaran S, Joo S., Zarghami S.,Hanifehpour Y., Samiei M., Koshki M., “Liposome: classification, preparation, and applications”, Nanoscale Research Letters 2013, 8:102 27 SONICATION drug solution in buffer to lipid mixture in 10 ml glass vial/ test tube. Sonicate the mixture with probe/ bath sonicator at 60 C for 3 minute.
  • 28. Mechanical Dispersion Method Dissolve lipids in organic solvent in a round bottom flask. Remove the solvent at 20 C using vacuum. Thin layer of lipids is formed on the edges. Add water at 0-60 C to the film to form MLVs. Akbarzadeh A., Sadabady R, Davaran S, Joo S., Zarghami S.,Hanifehpour Y., Samiei M., Koshki M., “Liposome: classification, preparation, and applications”, Nanoscale Research Letters (2013) 8:102 28
  • 29. MICROFLUIDIZER LIPOSOMES PREPARATION Lipid solution in organic solvent & aqueous drug solution are fluidized and allowed to interact at ultra high velocities within the interaction chamber at predefined conditions. As a result, homogenous vesicles are formed. Akbarzadeh A., Sadabady R, Davaran S, Joo S., Zarghami S.,Hanifehpour Y., Samiei M., Koshki M., “Liposome: classification, preparation, and applications”, Nanoscale Research Letters 2013, 8:102 29
  • 30. EXTRUSION TECHNIQUES Lipid mixture is converted into a thin film. The thin film is hydrated with aqueous drug solution. The resultant suspension is extruded through polycarbonate membrane as many as 8 times. Akbarzadeh A., Sadabady R, Davaran S, Joo S., Zarghami S.,Hanifehpour Y., Samiei M., Koshki M., “Liposome: classification, preparation, and applications”, Nanoscale Research Letters (2013), 8:102 30
  • 31. ETHANOL & ETHER INJECTION Dissolve surfactant in ether/ ethanol, introduce the solution in aqueous phase at 60º C through 14 gauge needle. The solvent will evaporate & SLVs are obtained. Akbarzadeh A., Sadabady R, Davaran S, Joo S., Zarghami S.,Hanifehpour Y., Samiei M., Koshki M., “Liposome: classification, preparation, and applications”, Nanoscale Research Letters 2013, 8:102 31
  • 32. Akbarzadeh A., Sadabady R, Davaran S, Joo S., Zarghami S.,Hanifehpour Y., Samiei M., Koshki M., “Liposome: classification, preparation, and applications”, Nanoscale Research Letters 2013, 8:102 32 liposomes resulting viscous suspension is diluted with PBS and heated on a water bath at 60°C for 10 min The organic phase is removed at 40°C under low pressure sonicated after addition of a small amount of phosphate buffered saline (PBS Sonicate the resulting two phase mixture at 4-5°C. The clear gel formed, Add the aqueous phase to the organic phase. Dissolve cholesterol and surfactant (1:1) in a mixture of ether and chloroform. Dissolve drug in water. Reverse phase evaporation method
  • 33. Detergent Solubilize phospholipid Removal of detergent Increased amount of Phospholipid Formation of LUVs Removal of detergent by dialysis . Detergent Removal Method Akbarzadeh A., Sadabady R, Davaran S, Joo S., Zarghami S.,Hanifehpour Y., Samiei M., Koshki M., “Liposome: classification, preparation, and applications”, Nanoscale Research Letters 2013, 8:102 33
  • 34. Therdphapiyana N., Jaturanpinyo M,Waranuch N.,Kongkaneramit L., Sarisuta N.,” Development and assessment of tyrosinase inhibitory activity of liposomes of Asparagus racemosus extracts”, Asian journal of pharmaceutical sciences 8 (2013)13 4 -1 42 34 Development and assessment of tyrosinase inhibitory activity of liposomes of Asparagus racemosus extracts Case study-1 Material •Phosphatidylcholine (PC90G) • Lecithin (LEC) •Cholesterol (CHOL), • Tert-butanol (TBN) •Diosgenin (DG) •.Kojic acid, •3,4-dihydroxy-L-phenylalanine (L-DOPA), • mushroom tyrosinase Chloroform-film method (CF) Reverse-phase evaporation method (REV) Polyol dilution method (PD) Freeze-drying of monophase solution method (MFD) Method of preparation OBJECTIVE: To develop liposomal formulation of asparagus racemosus root extract and evaluate physiochemical property and tyrosinase inhibitory activity
  • 35. Therdphapiyana N., Jaturanpinyo M,Waranuch N.,Kongkaneramit L., Sarisuta N.,”Development and assessment of tyrosinase inhibitory activity of liposomes of Asparagus racemosus extracts”, Asian journal of pharmaceutical sciences8 (2013)8,13 4 -1 42 35 Evaluation parameter •Microscopic appearance By Transmission electron microscopy [TEM] Observation: Concentric lamellar structure of vesicles. Structure method Multilamellar CF(A) MFD(B) Oligolamellar Structure REV (A) PD(c) •Particle size- 0.2 to 13.9um •Entrapment efficiency •Total saponin content •Tyrosinase inhibitory activity
  • 36. Therdphapiyana N., Jaturanpinyo M,Waranuch N.,Kongkaneramit L., Sarisuta N.,” Development and assessment of tyrosinase inhibitory activity of liposomes of Asparagus racemosus extracts”, Asian journal of pharmaceutical sciences (2013)8,13 4 -1 42 36
  • 37. Therdphapiyana N., Jaturanpinyo M,Waranuch N.,Kongkaneramit L., Sarisuta N.,” Development and assessment of tyrosinase inhibitory activity of liposomes of Asparagus racemosus extracts”, Asian journal of pharmaceutical sciences (2013)8,13 4 -1 42 37 •Tyrosinase inhibitory activity
  • 38. Therdphapiyana N., Jaturanpinyo M,Waranuch N.,Kongkaneramit L., Sarisuta N.” Development and assessment of tyrosinase inhibitory activity of liposomes of Asparagus racemosus extracts”, Asian journal of pharmaceutical sciences (2013)8,13 4 -1 42 38 conclusion Parameter result Size range 026-13.83um Zeta potential (charge) -1.5 -39.3mV Entrapment efficiency 42.19% -69.08% IN-VITRO TYROSINASE INHIBITORY ACTIVITY 12.06%-25.00% AR1-6 liposomes with LEC or PC90G as structural lipid could be prepared by CF, REV, PD, and MFD methods Types of lipid and preparation methods significantly influenced the physicochemical properties of liposomes such as vesicle type, size, surface charge, drug entrapment, and biological activity.
  • 39. . Ai-Hua Wen, Min-Koo Choi , and Dae-Duk Kim,” Formulation of Liposome for Topical Delivery of Arbutin” Arch Pharm Res (2006) (29) 12, 1187-1192 39 Formulation of Liposome for Topical Delivery of Arbutin Case study-3 Soybean phosphatidylcholine Cholesterol (CH)  Arbutin . Material Method LIPOSOMES :chloroform film method with sonication
  • 40. . . Ai-Hua Wen, Min-Koo Choi , and Dae-Duk Kim,” Formulation of Liposome for Topical Delivery of Arbutin” Arch Pharm Res (2006) (29) 12, 1187-1192 40
  • 41. . Ai-Hua Wen, Min-Koo Choi , and Dae-Duk Kim,” Formulation of Liposome for Topical Delivery of Arbutin” Arch Pharm Res (2006) (29) 12, 1187-1192 41
  • 42. . Ai-Hua Wen, Min-Koo Choi , and Dae-Duk Kim,” Formulation of Liposome for Topical Delivery of Arbutin” Arch Pharm Res (2006) (29) 12, 1187-1192 42 Conclusion  Liposomes containing AR, a skin-whitening agent, was successfully prepared by using the film dispersion method.  When these liposomes were applied occlusively to the mouse skin, the amount of AR permeated into the skin decreased when AR was encapsulated into liposome, while the enhanced accumulation of AR in the epidermis/dermis layers was found in the liposomal formulation. These findings might help to optimize targeting of arbutin ,Creating new opportunity for topical application
  • 43. Costin G., Trif M.,Nichita N., Dwek R.,and Petrescua S.,” pH-sensitive liposomes are efficient carriers for endoplasmic reticulum-targeted drugs in mouse melanoma cells”, Biochemical and Biophysical Research Communications (2002) 293,918–923 43 pH-sensitive liposomes are efficient carriers for endoplasmic reticulum-targeted drugs in mouse melanoma cells Case study-3 Material Cholesteryl hemisuccinate(chems) Cholestrol(chol) Dioleylphosphotidylethanolamine Phosphatidylcholine (PC), phosphatidylethanolamine (PE), And phosphatidylserine (PS) NB-DNJ, deoxynojirimycin (DNJ) and nonyl-dnj (Nn-dnj ) Cell culture. B16-F1 mouse melanoma cells Method Sonication method
  • 44. Costin G., Trif M.,Nichita N., Dwek R.,and Petrescua S.,” pH-sensitive liposomes are efficient carriers for endoplasmic reticulum-targeted drugs in mouse melanoma cells”, Biochemical and Biophysical Research Communications (2002) 293,918–923 44
  • 45. Costin G., Trif M.,Nichita N., Dwek R.,and Petrescua S.,” pH-sensitive liposomes are efficient carriers for endoplasmic reticulum-targeted drugs in mouse melanoma cells”, Biochemical and Biophysical Research Communications (2002) 293,918–923 45
  • 46. Costin G., Trif M.,Nichita N., Dwek R.,and Petrescua S.,” pH-sensitive liposomes are efficient carriers for endoplasmic reticulum-targeted drugs in mouse melanoma cells”, Biochemical and Biophysical Research Communications (2002) 293,918–923 46 The use of pH-sensitive liposomes composed of dioleoylphosphatidylethanolamine and cholesteryl hemisuccinate for the deliveryof NB-DNJ reduced the required dose for tyrosinase inhibition by a factor of 1000. The results indicate that these pH-sensitiveliposomes are efficient carriers for imino-sugars delivery in the endoplasmic reticulum of mammalian cells. encapsulation of glycosylation inhibitors in liposomes and their delivery inside the cells are promising for their future therapeutic use in pigmentation disorders Conclusion
  • 47. 47 Liposome –vesicular drug delivery system ameliorative tool for •Deliver drug at target site by avoiding its absorption at systemic circulation •Reducing its Frequency and Dose of application This characteristic make Liposome potential drug carrier system for treatment of skin pigmentation.
  • 48. REFERENCES: 1) Singhal M., Khanna S, Nasa A.,” COSMECEUTICALS FOR THE SKIN: AN OVERVIEW” Asian Journal of Pharmaceutical and Clinical Research, 2, (2011)1-6 2) Shoukat Parvez, Moonkyu Kang, Hwan-Suck Chung, Chongwoon Cho,Moo-Chang Hong, Min-Kyu Shin and Hyunsu Bae,” Survey and Mechanism of Skin Depigmenting and Lightening Agents”, PHYTOTHERAPY RESEARCH (2006) 20, 921–934. 3) Nina G. Jablonski and George Chaplin,” Human skin pigmentation, migration and disease susceptibility,” Phil. Trans. R. Soc. B (2012) 367, 785–792. 4) Hideaki Matsuda, Kazuya Murata, Kimihisa Itoh, Megumi Masuda and Shunsuke Naruto,” Melanin Hyperpigmentation Inhibitors from Natural Resources”’ Advances in Malignant Melanoma – Clinical and Research Perspectives ,170-184. 5) Le´ocadie Kamagaju, RenatoMorandini , EliasBizuru , PolycarpeNyetera , Jean BaptisteNduwayezu , CarolineSte´vigny , GhanemGhanem , PierreDuez ,” Tyrosinase modulation by five Rwandese herbal medicines traditionally used for skin treatment“,Journal of Ethnopharmacology (2013)146, 824–834 6) Shilimkar Vaibhav and K. Lakshaman, Tyrosinase Enzyme Inhibitory Activity of selected Indian Herbs,” International Journal of Research in Pharmaceutical and Biomedical Sciences (2008),2229-3701. 48
  • 49. References: 7) Tarl W. Prow, Jeffrey E. Grice,Lynlee L. Lin , Rokhaya Faye ,Margaret Butler,Wolfgang Becker ,Elisabeth M.T. Wurm , Corinne Yoong , Thomas A. Robertson , H. Peter Soyer , Michael S. Roberts,” Nanoparticles and microparticles for skin drug delivery”, Advanced Drug Delivery Reviews (2011)63, 470–491 8) Theresa M. Allen a,⁎, Pieter R.Cullis,” Liposomal drug delivery systems: From concept to clinical applications”, Advanced Drug Delivery Reviews (2013) 65,36–48. 9) . Martinho N.,Damge C., Reis C. ,”Recent Advances in drug Delivery Systems”, Journal of Biomaterials and Nanobiotechnology, (2011), 2, 510-526. 10) . Sharma A, Sharma U., “Liposomes in drug delivery: progress and limitations”, International Journal of Pharmaceutics (1997)154, 123 -140. 11) Kulkarni P., Yadav J., Vaidya K., “LIPOSOMES: A NOVEL DRUG DELIVERY SYSTEM”, International Journal of Current Pharmaceutical Research, (2011)4(3) 10-18 12) Akbarzadeh A., Sadabady R, Davaran S, Joo S., Zarghami S.,Hanifehpour Y., Samiei M., Koshki M., “Liposome: classification, preparation, and applications”, Nanoscale Research Letters 2013, 8,102 49
  • 50. 50 References 16. Therdphapiyana N., Jaturanpinyo M,Waranuch N.,Kongkaneramit L., Sarisuta N.,” Development and assessment of tyrosinase inhibitory activity of liposomes of Asparagus racemosus extracts”, Asian journal of pharmaceutical sciences (2013)8,13 4 -1 42 17. Ai-Hua Wen, Min-Koo Choi , and Dae-Duk Kim,” Formulation of Liposome for Topical Delivery of Arbutin” Arch Pharm Res,2006( 29) 12, 1187-1192 18.Costin G., Trif M.,Nichita N., Dwek R.,and Petrescua S.,” pH-sensitive liposomes are efficient carriers for endoplasmic reticulum-targeted drugs in mouse melanoma cells”, Biochemical and Biophysical Research Communications (2002)213, 918–923
  • 51. 51