Niosomes a novel drug delivery system

1. NIOSOMES: A NOVEL DRUG DELIVERY SYSTEM Presented by: Sanjay Kumar Yadav Enrollment No: A10647013015 Amity Institute of Pharmacy (AIP)

2. Introduction Factors Affecting Niosomes Preparation Methods of Preparation Characterization of Niosomes Stability of Niosomes Applications of Niosomes Toxicity of Niosomes PRESENTATION FLOW

3. NOVEL DRUG DELIVERY SYSTEM (NDDS ) Refers to approaches, formulations, technologies, and systems for transporting a pharmaceutical compound in the body as needed to safely achieve its desired  therapeutic effect May involve scientific site-targeting within the body, or facilitating systemic pharmacokinetics Technologies modify drug release profile, absorption, distribution and elimination for the benefit of Improving product efficacy and safety Patient convenience and compliance INTRODUCTION

4. EXAMPLES OF NDDS Niosomes Liposomes Nanoparticles Resealed erythrocytes Microspheres Monoclonal antibodies Micro emulsions Antibody-loaded drug delivery Magnetic microcapsules Implantable pumps Figure 1: various drug delivery systems ( Aitha S, 2013)

5. Novel drug delivery system, in which the medication is encapsulated in a vesicle which is composed of a bilayer of non-ionic surface active agents ( Nasir A, 2012 ) Are very small, and microscopic in size. Although structurally similar to liposomes, they offer several advantages over them. NIOSOMES Figure 2: Niosomes Vesicles ( Aitha S, 2013)

6. The vesicles forming amphiphile is a non-ionic surfactant stabilized by addition of cholesterol and small amount of anionic surfactant such as dicetyl phosphate NIOSOMES Figure 3: Vesicle of niosome ( Aitha S, 2013)

7. Figure 4: Structure of Niosomes STRUCTURE OF NIOSOMES similar to liposomes, in that they are also made up of a bilayer. However, the bilayer in the case of Niosomes is made up of non-ionic surface active agents rather than phospholipids. Made of a surfactant bilayer with its hydrophilic ends exposed on the outside and inside of the vesicle, while the hydrophobic chains face each other within the bilayer. (Patel SM et al, 2012 ) ( Makeshwar KB, 2013)

8. STRUCTURE OF NIOSOMES vesicle holds hydrophilic drugs within the space enclosed in the vesicle, while hydrophobic drugs are embedded within the bilayer itself. Niosomes vesicle would consist of a vesicle forming amphiphile i.e. a non-ionic surfactant such as Span- 60, which is usually stabilized by the addition of cholesterol ( Makeshwar KB, 2013 ) Figure 5: Structure of niosome ( Makeshwar KB, 2013 )

9. Entrap solutes in a manner analogous to liposomes . Osmotically active and stable. Accommodate the drug molecules with a wide range of solubility. Exhibits flexibility in their structural characteristics (composition, fluidity and size ) Performance of the drug molecules is increased. Better availability to the particular site by protecting the drug from biological environment. Surfactants used in preparation are biodegradable, biocompatible and non-immunogenic SALIENT FEATURES OF NIOSOMES ( Makeshwar KB, 2013)

10. Improve the therapeutic performance of the drug molecules by Delayed clearance from the circulation Protecting the drug from biological environment Restricting effects to target cells Niosomal dispersion in an aqueous phase can be emulsified in a nonaqueous phase to Regulate the delivery rate of drug Administer normal vesicle in external non-aqueous phase. Handling and storage of surfactants requires no special conditions. Bioavailability of poorly absorbed drugs is increased. Targeted to the site of action by oral, parenteral as well as topical routes. ADVANTAGES OF NIOSOMES DELIVERY SYSTEM ( Makeshwar KB, 2013)

11. According to the nature of lamellarity Multilamellar vesicles (MLV) 1-5 μm in size. Large Unilamellar vesicles (LUV) 0.1 – 1μm in size Small Unilamellar vesicles (SUV) 25 – 500 nm in size. According to the size Small Niosomes (100 nm – 200 nm) Large Niosomes (800 nm – 900 nm) Big Niosomes (2 μm – 4 μm) TYPES OF NIOSOMES

12. FACTORS AFFECTING THE FORMATION OF NIOSOMES

13. Type of surfactant influences encapsulation efficiency, toxicity, and stability of Niosomes Mean size of Niosomes increases proportionally with increase in the HLB of surfactants N ATURE OF SURFACTANT

14. The surfactant/lipid ratio is generally 10-30 mM (1-2.5% w/w) Increasing the surfactant/lipid level increases the total amount of drug encapsulated SURFACTANT AND LIPID LEVELS

15. NATURE OF THE DRUG The Physio -chemical properties of encapsulated drug influence charge and rigidity of the Niosome bilayer. The drug interacts with surfactant head groups and develops the charge that creates mutual repulsion between surfactant bilayers, and hence increases vesicle size . The aggregation of vesicles is prevented due to the charge development on bilayer. Table 1: Effect of the nature of drug on formation vesicle ( Kazi KM et al, 2010)

16. CHOLESTEROL ( Tamizharas S et al, 2009) Addition of cholesterol molecule to Niosomal system Makes the membrane rigid Reduces leakage of drug from the Niosome Increases the chain order of bilayer Strengthen the non‑polar tail of the non‑ionic surfactant Increase in the entrapment efficiency Leads to the transition from the gel state to liquid phase in Niosomes systems MEMBRANE ADDITIVES Cholesterol

17. Charge inducers are one of the membrane additives which are often included in Niosomes because Increase surface charge density Prevent vesicles flocculation, Aggregation and Fusion. Examples: Dicetyl phosphate (DCP) and Stearyl amine ( SA ) MEMBRANE ADDITIVES ( Nasir A, 2012 )

18. Film Method Ether Injection Method Sonication Reverse Phase Evaporation Heating Method Microfluidization Multiple Membrane Extrusion Method Transmembrane pH gradient (inside acidic) Drug Uptake Process (remote Loading) The “Bubble” Method Formation of Niosomes from Proniosomes METHODS OF PREPARATION ( Madhav NVS, 2011 )

19. FILM METHOD Also known as hand shaking method

20. FILM METHOD Figure 6: Steps of Film method ( Madhav NVS, 2011)

21. ETHER INJECTION METHOD Figure 7: Steps of Ether injection method ( Madhav NVS, 2011)

22. SONICATION Figure 8: Sonication method ( Madhav NVS, 2011)

23. REVERSE PHASE EVAPORATION

24. Non-toxic, Scalable and one-step method. HEATING METHOD

25. Recent technique used to prepare Unilamellar vesicles of defined size distribution. based on submerged jet principle MICROFLUIDIZATION

26. MICROFLUIDIZATION Figure 9: Steps of microfludization method ( Madhav NVS, 2011)

27. Good method for controlling Niosomes size. MULTIPLE MEMBRANE EXTRUSION METHOD Figure 10: Multiple membrane extrusion method ( Madhav NVS, 2011)

28. TRANSMEMBRANE pH GRADIENT DRUG UPTAKE PROCESS

29. A recently developed technique which allows the preparation of Niosomes without the use of organic solvents. BUBBLE METHOD

30. FORMATION OF NIOSOMES FROM PRONIOSOMES ( Makeshwar KB, 2013) T=Temperature. Tm = mean phase transition temperature

31. POST-PREPARATION PROCESSES

32. 1) Dialysis: The aqueous niosomal dispersion is dialyzed in a dialysis tubing against phosphate buffer or normal saline or glucose solution. 2) Gel Filtration: The unentrapped drug is removed by gel filtration of niosomal dispersion through a Sephadex -G -50 column and elution with phosphate buffered saline or normal saline. 3) Centrifugation: The niosomal suspension is centrifuged and the supernatant is separated. The pellet is washed and then resuspended to obtain a niosomal suspension free from unentrapped drug. POST-PREPARATION PROCESSES ( Makeshwar KB, 2013)

33. Size, Shape and Morphology Entrapment efficiency Vesicle diameter In vitro release Vesicle charge Bilayer rigidity and Homogeneity Osmotic Shrinkage Physical stability of vesicles at different temperature Turbidity Measurement CHARACTERIZATION OF NIOSOMES

34. Structure of surfactant based vesicles has been visualized and established using freeze fracture microscopy Photon correlation spectroscopy used to determine mean diameter of the vesicles . Electron microscopy used for morphological studies of vesicles Laser beam is generally used to determine size distribution, mean surface diameter and mass distribution of Niosomes. SIZE, SHAPE AND MORPHOLOGY

35. After preparing Niosomal dispersion , unentrapped drug is separated by Dialysis Centrifugation Gel filtration Drug remained entrapped in Niosomes is determined by complete vesicle disruption using 50% n-propanol or 0.1% Triton X-100 and analysing the resultant solution by appropriate assay method for the drug . (Bragagnia M, 2012) ENTRAPMENT EFFICIENCY

36. To determine drug loading and encapsulation efficiency , the niosomal aqueous suspension was ultracentrifuged , supernatant was removed and sediment was washed twice with distilled water in order to remove the adsorbed drug . The Niosomal recovery was calculated as: NIOSOMAL DRUG LOADING ( Makeshwar KB, 2013 )

37. Niosomes diameter can be determined using Light microscopy Photon correlation microscopy Freeze fracture electron microscopy . Freeze thawing VESICLE DIAMETER ( Shirsand SB, 2012) Figure 11: Microphotograph of niosomes ( Shrisand SB, 2012)

38. IN VITRO RELEASE ( Makeshwar KB, 2013 )

39. The vesicle surface charge can play an important role in the behaviour of Niosomes in vitro and in vivo . Charged Niosomes are more stable against aggregation and fusion than uncharged vesicles . In order to obtain an estimate of the surface potential, the zeta potential of individual Niosomes can be measured by Microelectrophoresis, Fluorophores, and Dynamic light scattering. Zeta potential is calculated by using Henry equation (S P Vyas, 2011 ) Where is Zeta potential, is electrophoretic mobility, is viscosity of the medium and is dielectric constant   VESICLE CHARGE ( Makeshwar KB, 2013)

40. The biodistribution and biodegradation of Niosomes are influenced by rigidity of the bilayer . Homogeneity can occur both within Niosomes structures themselves and between Niosomes in dispersion and could be identified via . NMR , Differential Scanning Calorimetry ( DSC) and Fourier transform-infra red spectroscopy (FT-IR) techniques. Membrane rigidity can be measured by means of mobility of fluorescence probe as a function of temperature . (Patel SM et al, 2012) BILAYER RIGIDITY AND HOMOGENEITY

41. Osmotic shrinkage of vesicles can be determined by monitoring reductions in vesicle diameter, initiated by addition of hypertonic salt solution to suspension of Niosomes. Niosomes prepared from pure surfactant are osmotically more sensitive in contrast to vesicles containing cholesterol. OSMOTIC SHRINKAGE

42. Aggregation or fusion of vesicles as a function of temperature was determined as the changes in vesicle diameter by laser light scattering method . The vesicles were stored in glass vials at room temperature or kept in refrigerator (4 o C) for 3 months. The changes in morphology of Multilamellar vesicles (MLVs) and also the constituent separation were assessed by an optical microscope. The retention of entrapped drug were measured 72 hours after preparation and after 1, 2 or 3 months in same formulations PHYSICAL STABILITY OF VESICLES AT DIFFERENT TEMPERATURE

43. Niosomes were diluted with bidistilled water to give a total lipid concentration of 0.312 mM After rapid mixing by sonication for 5 min Turbidity was measured as the absorbance with an ultraviolet-visible diode array spectrophotometer. TURBIDITY MEASUREMENT

44. Vesicles are stabilized based upon formation of 4 different forces: 1. Van der Waals forces among surfactant molecules 2. Repulsive forces emerging from the electrostatic interactions among charged groups of surfactant molecules 3. Entropic repulsive forces of the head groups of surfactants 4. Short-acting repulsive forces. STABILITY OF NIOSOMES

45. FACTORS AFFECTING STABILITY OF NIOSOMES

46. A surfactant used for preparation of Niosomes must have a hydrophilic head and hydrophobic tail. The hydrophobic tail may consist of one or two alkyl or perfluoroalkyl groups or in some cases a single steroidal group. The ether type surfactants with single chain alkyl as hydrophobic tail is more toxic than corresponding dialkylether chain. The ester type surfactants are chemically less stable than ether type surfactants and the former is less toxic than the latter due to ester-linked surfactant degraded by esterases to triglycerides and fatty acid in vivo. The surfactants with alkyl chain length from C12-C18 are suitable for preparation of Niosome. NATURE OF SURFACTANT (Singh CH, 2011)

47. The geometry of vesicle to be formed from surfactants is affected by its structure, which is related to critical packing parameters Critical packing parameters can be defined using following equation , Where v = hydrophobic group volume, lc = the critical hydrophobic group length a0 = the area of hydrophilic head group From the critical packing parameter value type of miceller structure formed can be ascertained as given below, If CPP < ½ then formation of spherical micelles , If ½ < CPP < 1 formation of bilayer micelles, If CPP > 1 formation inverted micelles23 . surfactants with longer alkyl chains generally give larger vesicles   STRUCTURE OF SURFACTANT ( Madhav NVS, 2011 )

48. The physico -chemical properties of encapsulated drug influence charge and rigidity of the Niosome bilayer. The drug interacts with surfactant head groups and develops the charge that creates mutual repulsion between surfactant bilayers and hence increases vesicle size. NATURE OF ENCAPSULATED DRUG (Singh CH, 2011)

49. Hydration temperature influences the shape and size of the Niosome. For ideal condition it should be above the gel to liquid phase transition temperature of system . Temperature change of Niosomal system affects assembly of surfactants into vesicles and also induces vesicle shape transformation TEMPERATURE OF HYDRATION ( Madhav NVS, 2011)

50. Niosomes as Drug Carriers Diagnostic imaging with Niosomes Drug Targeting Delivery to the brain Anti cancer drugs Anti infectives Targeting of bioactive agents To Reticulo -endothelial system (RES) To organs other than RES NIOSOME DELIVERY APPLICATIONS ( Malhotra M et al, 1994 )

51. Ophthalmic drug delivery Delivery of peptide drugs Immunological application of Niosomes Transdermal delivery of drugs by Niosomes Delivery system for the vasoactive intestinal peptide (VIP ) Niosomes as carriers for Hemoglobin Niosomal vaccines NIOSOME DELIVERY APPLICATIONS

52. Sustained Release Localized Drug Action OTHER APPLICATIONS

53. Unfortunately, there is not enough research conducted to investigate toxicity of Niosomes. It was determined that the ester type surfactants are less toxic than ether type surfactants. In general, the physical form of Niosomes did not influence their toxicity as evident in a study comparing the formulations prepared in the form of liquid crystals and gels . Nasal applications of these formulations caused toxicity in the case of liquid crystal type Niosomes. TOXICITY OF NIOSOMES

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