FORMULATION AND EX-VIVO EVALUATION somea

FORMULATION AND EX-VIVO EVALUATION somea

1. FORMULATION AND EX-VIVO EVALUATION OF NIOSOMAL SUSPENSION OF ABIRATERONE ACETATE   Project Thesis submitted to Andhra University In partial fulfillment of the requirement for the award of degree of Bachelor of Pharmacy N. TIRUPATHI RAO ( Reg. No: 621150901014) M.LALITHA ( Reg. No:621150901013) L.AMRUTHA ( Reg. No:621150901012) V.DEVI ( Reg. No:621150901021) Under the esteemed guidance of Dr. K.L. DEEPTHI, M. PHARM, Ph.D., PROFESSOR, DEPT. OF PHARMACEUTICS SRI SAI COLLEGE OF PHARMACY Approved by Pharmacy Council of India, Affiliated to Andhra University, Since: 2007-2008 .

2. INTRODUCTION Niosomes are the unique bilayer lipid nanostructure developed with help of the self-aggregation of a non- ionic surfactant, initially, it has been developed by the L Oreal company for its cosmetics application in 1975. Niosomes can be alternative to liposomes and polymersomes duo to their ability to encapsulate different types of drugs for increasing stability and efficacy. Niosomes are structurally similar to liposomes, but they have greater advantages over liposomes.one of this advantage is that niosomes are considered to be more stable when compared to liposomes. Niosomes are mix of cholesterol and non-ionic surfactants. CLASSIFICATION OF NIOSOMES Niosomes are classified based on the number and size of vesicular lipid layers. Based on this classification, two types. Unilamellar vesicles Multi-lamellar vesicles The unilamellar vesicles are classified into two types. Small unilamellar vesicles (SUV) Large unilamellar vesicles (LUV) 1

3. COMPONENTS OF NIOSOMES NON-IONOC SURFACTANTS Non-ionic surface-active agents are used to preparation of niosomes. They are amphiphilic molecules with apolar head and a non- polar tail. These two parts are usually connected by ether, ester, bonds or amide bonds. The hydrophilic and lipophilic balance of the surfactant determines the formation of the bilayer structure of vesicles. CHOLESTEROL Cholesterol is a white waxy solid steroid, an amphiphilic molecule, responsible for the rigidity, fluidity, permeability, and efficacy of encapsulation in niosome composition. Niosome vesicle’s structure can be affected by cholesterol because the stability of bilayers can be enhanced through the formation of hydrogen bonds between hydroxyl groups and the alkyl chains of the surfactant molecules 2

4. AIM & OBJECTIVE Aim To formulation and Ex-vivo evaluation of niosomal suspension of abiraterone acetate Objectives Preparation & Ex-vivo evaluation of niosomes. To formulate Abiraterone acetate in niosomal suspension.. The need of present study is to encapsulate the drug in niosomes vesicles for effective drug delivery system. Criteria for selection of drug As Abiraterone acetate is a BCS class IV drug, it is suitable for the preparation of niosomes. Abiraterone acetate is an anti-cancer drug used to treat metastatic castration-resistant prostate cancer and hormone-sensitive high-risk metastatic prostate cancer. As abiraterone has poor oral bioavailability and is susceptible to hydrolysis by esterases, abiraterone acetate was developed as an orally bioavailable prodrug with enhanced stability and absorption 3

5. PLAN OF WORK SELECTION OF DRUG Abiraterone acetate is an anti-cancer drug used to treat metastatic castration-resistant prostate cancer and hormone-sensitive high-risk metastatic prostate cancer. Abiraterone acetate is a BCS class IV drug. SELECTION OF FORMULATION As Abiraterone acetate is a BCS class IV drug, it is suitable for the preparation of niosomes. SELECTION OF METHOD Ether injection method is used for preparation of niosomal suspension of abiraterone acetate SELECTION OF POLYMERS & EXCIPIENT & SOLVENTS Niosomes are small sized vesicular system containing components of non-ionic surfactants and cholesterol. In this research work selecting the non-ionic surfactants like Tween 80 and Span 60. Cholesterol is used to preparation of niosomal suspension by using ether injection method. Ethanol and diethyl ether are used in this research formulation 4

6. LITERATURE REVIEW 1.Xin Liu et al They studied about the ph. sensitive Niosomes which are designed to specifically triggered release the drugs in response to the change of pH in the surrounding serum. pH-sensitive Niosomes are designed to specifically triggered release the drugs in response to the change of pH in the surrounding serum. So, pH-sensitive Niosomes can effectively deliver drug or gene fragments into the cytoplasm via the endocytosis pathway. 2 . D. DEAMER et al Niosomes, in the size range of microsomal vesicles, are formed when ether solutions of a variety of lipids are injected into warm aqueous solutions. The Niosomes are osmotically active, most are unilamellar, and the volume trapping efficiency is approximately ten times that of sonicated and hand-shaken preparations. 3 .JOHN C MATHAI et al They studied about Preparation of Large UniLamellar Niosomes by the Ether Injection Method and Evaluation of the Physical Integrity by Osmometry. Phospholipids in aqueous suspension aggregate to form stable bilayers enclosing an aqueous space within. These structures are very similar to biological membranes except that they do not contain protein, and indeed one can now prepare Niosomes with protein embedded in the bilayer. 4.David W. Deamer He studies about preparation and properties of ether injection Niosomes. There are a number of different methods now available for producing Niosomes. Each preparation method has certain features that make it suitable for some studies and unsuitable for others 5

7. DRUG PROFILE Abiraterone is a potent, irreversible, and selective inhibitor of 17 αhydroxylase/C17,20-lyase (CYP17), an enzyme expressed in testicular, adrenal, and prostatic tumour tissues, to regulate androgen biosynthesis.2,7,9 Abiraterone was first approved by the FDA and EMA on April,7 July,14 and September 2011,7 respectively. It is used to treat metastatic castration-resistant prostate cancer and hormone-sensitive high-risk metastatic prostate cancer. As abiraterone has poor oral bioavailability and is susceptible to hydrolysis by esterases, abiraterone acetate was developed as an orally bioavailable prodrug with enhanced stability and absorption. Weight Average: 349.509 Chemical Formula : C 24 H 31 NO Polar surface area :39.19 A2 Refractivity :116.45 m.mol-1 Polarizability :46.53a3 Melting point :419K Solubility: Ethanol Tetra hydro furan Acetone Diethyl ether Dichloromethane Diethyl ether Acetonitrile Poor water soluble BCS Classification: IV Storage condition ;20’C 6

8. EXCIPIENT PROFILE 1.Cholesterol Cholesterol occurs as white or faintly yellow, almost odourless, pearly leaflets, needles, powder, or granules. On prolonged exposure to light and air, cholesterol acquires a yellow to tan colour Boiling point: 36.08C Density: 1.052 g/cm3 for anhydrous form. Dielectric constant D20: 5.41 Melting point: 147–1508C Specific rotation [a]D 20: -39.58 (2% w/v solution in chloroform); -31.58 (2% w/v solution in ether). Functional CATEGORIES Stabilizing of the niosomes Improve the encapsulation efficiency 2.Polysorbate 80 Polyoxyethylene sorbitan fatty acid esters (polysorbates) are a series of partial fatty acid esters of sorbitol and its anhydrides copolymerized with approximately 20, 5, or 4 moles of ethylene oxide for each mole of sorbitol and its anhydrides. Functional Category Emulsifying agent; non-ionic surfactant; solubilizing agent; wetting, dispersing/suspending agent 3. Sorbitan Esters (Sorbitan Fatty Acid Esters Sorbitan monoesters are a series of mixtures of partial esters of sorbitol and its mono- and dianhydrides with fatty acids. Sorbitan diesters are a series of mixtures of partial esters of sorbitol and its monohydride with fatty acids. Sorbitan esters are widely used in cosmetics, food products, and pharmaceutical formulations as lipophilic non-ionic surfactants. 7

9. METHODOLOGY Construction of calibration curve of abiraterone acetate by uv -visible Spectrometry λ max curve of abiraterone acetate To determine the optimum Amax of abiraterone acetate 100µg/ml of working standard solution was preferred and scanned in UV wavelength range of 200-400nm using HCL as a blank in the selected solvent drug show 234nm Amax as shown in fig16 Preparation of standerd stock solution Accurately weigh 100mg of abiraterone acetate transferred to 100ml of volumetric flask and add 30 ml of ethanol and make up into HCL buffer solution and labelled as standerd solution. Preparation of working solution 10ml of abiraterone acetate stock solution was pipetted out and taken in 100ml volumetric flask and make up into HCL buffer solution . 8

10. Formulation OF NIOSOMAL suspension of abiraterone acetate ETHER INJECTION METHOD Cholesterol and surfactants were taken in a different concentration and dissolved in diethyl ether. The previously drug dissolved in ethanol in a 100 ml beaker. 7.4 pH phosphate buffer in a 100 ml beaker were placed on a magnetic stirrer at temperature 60-65ºC and moderate rpm was maintained and stirred about 45 min. Non-ionic surfactant and cholesterol and 5ml diethyl ether Dissolved Drug and ethanol Dissolved Both solutions are mixed and injected using a 10cc needle Added to 15ml 7.4 phosphate buffer solution Formation of niosomes 9

11. EXCIPENTS F1 F2 F3 F4 F5 F6 F7 F8 ABIRATERONE ACETATE 250mg 250mg 250mg 250mg 250mg 250mg 250mg 250mg CHOLESTEROL 100mg 100mg 100mg 100mg 100mg 100mg 100mg 100mg DIETHYL ETHER 5ml 5ml 5ml 5ml 5ml 5ml 5ml 5ml ETHANOL 2.5ml 2.5ml 2.5ml 2.5ml 2.5ml 2.5ml 2.5ml 2.5ml SPAN 60 0.1ml 0.2ml 0.3ml 0.4ml         TWEEN 80 - - - - 0.1ml 0.2ml 0.3ml 0.4ml Formulation table of Niosomal suspension of abiraterone acetate 10

12. EVALUATION OF NIOSOMES PHYSICAL CHARACTERIZATION Optical microscopy Niosomal suspension was viewed by microscope at 100X and 45X magnification to observe the size and shape of the Niosomal particles. Particles are characteristics of suspension by spreading a thin layer of niosomal suspension on a slide and placing the coverslip on it. FORMULATION 1 250:0.1:0.1 FORMULATION 2 250:0.2:0.1 FORMULATION 3 250:0.3:0.1 FORMULATION 6 250:0.2:0.1 FORMULATION 7 250:0.3:0.1 FORMULATION 5 250:0.1:0.1 11

13. Ex-vivo permeability study by using chicken intestinal Sac Three cleaned chicken intestinal segments each of 16 cm length were ringed in PSS and in phosphate buffer (pH 7.4) 2 mL of niosomal suspension was placed in the intestine which was the sealed at both the Ends These segments were then placed in the Magnetic stirrer filled with 1000 ml of phosphate buffer (pH 7.4) at 100 rpm with a temperature of 37.0C. Each 10 ml of sample was withdrawn from each compartment at predetermined intervals (10,20,30,40,50, -----240 Min). Permeability study was checked for 4 hours and absorbance was taken at 234nm by using U.V &Visible spectrophotometer 12

14. RESULTS & DISCUSSION λ max curve of abiraterone acetate Determined the optimum Amax of abiraterone acetate 100µg/ml of working standard solution was preferred and scanned in UV wavelength range of 200-400nm using HCL as a blank in the selected solvent drug show 234nm as shown in fig Preparation of standard curve CONCENTRATION(µg/ml) ABSORBANCE AT 234 nm 10 0.128 20 0.222 40 0.394 60 0.587 80 0.744 100 0.932 13

15. Ex-vivo permeability study by using chicken intestinal sac Time(min) Absorbance % Permeability 30 0.122 8% 60 0.186 16% 90 0.258 24% 120 0.319 31% 150 0.471 48% 180 0.611 64% 210 0.697 74% 240 0.711 75% Absorbance % Permeability 0.106 7% 0.140 10% 0.213 19% 0.318 30% 0.425 42% 0.529 54% 0.638 66% 0.708 74% FORMULATION 7 FORMULATION 3 14

16. % PERMEABILITY OF NIOSOMAL SUSPENSION In this Ex-vivo permeation study of niosomal suspension of abiraterone acetate showed that niosomal suspension formulation 3 has a higher rate of permeation compared to formulation 7. The increased rate of permeation from the abiraterone acetate niosomal suspension may be due to the permeation enhancing effects of niosomes. 15

17. SUMMARY To determine the optimum Amax of abiraterone acetate 100µg/ml of working standard solution was preferred and scanned in UV wavelength range of 200-400nm using HCL as a blank in the selected solvent drug show 234nm Evaluation of Abiraterone acetate Niosomes: Various parameters like optical microscopy were done for niosomes. Niosomes were subjected to Ex-vivo permeability studies. It has been observed that the F3 formulation possessed 75% permeability. Drug loading has efficiently been done where highest frequency of permeation has occurred. Niosomal suspension was viewed by microscope at 100X and 45X magnification to observe the size and shape of the Niosomal particles. Particles are characteristics of suspension by spreading a thin layer of niosomal suspension on a slide and placing the coverslip on it. In this Ex-vivo permeation study of niosomal suspension of abiraterone acetate showed that niosomal suspension formulation 3 has a higher rate of permeation compared to formulation 7. The increased rate of permeation from the abiraterone acetate niosomal suspension may be due to the permeation enhancing effects of niosomes. 16

18. CONCLUSION Abiraterone acetate niosomal suspension was prepared by using ether injection method and it was evaluated for ex-vivo permeation studies. From the results of optical microscopy, it was concluded that the formulation F3 and F7 was concluded as optimized formulations in my research work. In this formulation F3 and F7 we at gone for ex-vivo permeability study, among the results of permeability study the F3 formulation as gone maximum permeability after 4 hours permeability study. 17

19. REVIEW ARTICLE SUBMISSION I am writing to acknowledge the receipt of manuscript titled A REVIEW ARTICLE ΟΝ; ΝΑΝΟΤECHNOLOGY FOR COLON DRUG DELIVERY SYSTEM’ which you submitted to International Journal of Pharmaceutical Sciences and Research. Acknowledgement no: IJPSR/RE-7607/02-25 has been recommended for publication in upcoming Vol. 16, Issue 8; August 2025 Issue of International Journal of Pharmaceutical Sciences and Research. August issue will be available online from August 1st, 2025. 18

20. REFERENCES 1. Khan, R.; Irchhaiya, R. Niosomes: A potential tool for novel drug delivery. J. Pharm. Investig. 2016, 46, 1–10. 2.Malipeddi, V.R.; Dua, K.; Pinto, T.D.J.A.; Kikuchi, I.S.; Kulkarni, G.T.; Pant, I.; Awasthi, R. Opportunities and Challenges in Nano-structure Mediated Drug Delivery: Where Do We Stand? Curr. Nanomed. 2016, 6, 78–104 3.Marianecci, C.; Marzio, L.D.; Rinaldi, F.; Celia, C.; Paolino, D.; Alhaique, F.; Esposito, S.; Carafa, M. Niosomes from 80s to present: The state of the art. Adv. Colloid Interface Sci. 2014, 205, 187–206 4.Abtahi, N.A., Naghib, S.M., Ghalekohneh, S.J., Mohammadpour, Z., Nazari, H., Mosavi, S.M., Gheibihayat, S.M., Haghiralsadat, F., Reza, J.Z., Doulabi, B.Z., 2021. Multifunctional stimuli-responsive niosomal nanoparticles for co-delivery and co- administration of gene and bioactive compound: in vitro and in vivo studies. Chem. Eng. J. 429, 132090 5.Ag Seleci, D., Seleci, M., Walter, J.G., Stahl, F., Scheper, T., 2016. Niosomes as nanoparticular drug carriers: Fundamentals and recent applications. J. Nanomater. 2016. 19