chapter no. title page no.shodhganga.inflibnet.ac.in/bitstream/10603/9096/2... · 9. 4c.1...

iv

Table of Contents

Chapter No. Title Page No.

1 INTRODUCTION 1-10

1.1 Introduction to topic

1.2 Biopharmaceutics Classification system

1.3 Importance of bioavailability

1.4 Basics about bioavailability and absorption

1.5 Problems due to low bioavailability

1.6 Methods for enhancing bioavailability

1.7 Aims and objectives

2 LITERATURE REVIEW 11-43

2.1 Reported methods for enhancing solubility and dissolution of the drug

2.2 Selection of drug

2.3 Drug profile

2.4 Drug characterization

2.5 Formulation patents related to drug

2.6 Selection of techniques for solubility enhancement

3 RESEARCH ENVISAGED 44

4 EXPERIMENTAL 45-108

4.1 List of equipment’s used during project

work

4.2 List of materials used during project work

4A Preformulation of drug

4B Development of analytical method

4B.1 Ultraviolet spectroscopic (UV) method development for drug release

4B.2 Development of high performance liquid chromatographic (HPLC) method for assay

4C Solubility enhancement with cyclodextrins

4D Solubility enhancement using “liquisolid compacts” technology

4E Solubility enhancement using self microemulsifying drug delivery systems

v

Chapter No. Title Page No.

4F Solubility enhancement by preparation of nanoparticles

4F.1 Preparation of nanoparticles with supercritical Antisolvent precipitation

4F.2 Preparation of nanoparticles using ion gelation technique

4F.3 Preparation of nanoparticles using nanoencapsulation technique

4G Optimization of formulation

4H In –vivo studies

4H.1 Development and validation of analytical method for analysis of candesartan cilexitil in plasma (LC-MS/MS)

4H.2 In Vivo bioavailability of selected formulations in rats

4I Accelerated stability testing

5 RESULTS AND DISCUSSION 109-245

6 SUMMARY AND CONCLUSION 246-256

7 APPENDIX 257-258

8 REFERENCES 259-280

9 LIST OF RESEARCH OUTPUT 281

vi

Sr. No Table

No. Name of table

Page

No.

1. 1.1 GI Physiology and drug Absorption 8

2. 2.1 List of antihypertensive agents with their solubility and

bioavailability

22

3. 2.2 Characterization of solid forms of Candesartan cilexetil 36

4. 2.3 Crystal properties of Form I and Form II of

candesartan cilexetil

37

5. 2.4 Formulation patents related to drug 39-43

6. 4.1 List of equipments 45

7. 4.2 List of materials 46

8. 4B.1 Optimized chromatographic conditions for analysis of

candesartan cilexetil by HPLC method

50

9. 4C.1 Applications of cyclodextrins in marketed

pharmaceutical products

56

10. 4C.2 Effect of cyclodextrin complexation on various

classes of drugs in BCS classification

59

11. 4C.3 Formulations prepared with cyclodextrins 66

12. 4D.1 Excipients used for formulation of liquisolid compacts 69

13. 4D.2 Formulations with liquisolid technology 71

14. 4E.1 Example of surfactants, co-surfactant, and co-solvent

used in commercial formulations.

77

15. 4E.2 Excipients used for formulation of SMEDDS 84

16. 4E.3 Combinations for formulation of SMEDDS 85

17. 4F.1 Critical conditions for some solvents 89

18. 4F.2 Excipients used for formulation of nanoparticles by ion

gelation method

98

19. 4F.3 Formulation design for preparation of nanoparticles by

ion gelation method

98

20. 4F.4 Excipients used for formulation of nanoparticles by

nanoencapsulation method

101

21. 4F.5 Formulation design for preparation of nanoparticles by

nanoencapsulation method

102

22. 5A.1 Physicochemical characterization of drug 109

23. 5A.2 pH solubility profile of drug 112

24. 5A.3 Percentage cumulative release of drug in multimedia

dissolution

112

25. 5A.4 Percentage cumulative release of marketed formulation

in multimedia dissolution

113

26. 5B.1 Precision of UV method 118

27. 5B.2 Accuracy of UV method 119

28. 5B.3 System precision of HPLC method 120

LIST OF TABLES

vii

29. 5B.4 Linearity of HPLC method 124

30. 5B.5 Limit of detection and Limit of quantification of CC by

HPLC method

125

31. 5B.6 Accuracy of HPLC method 126

32. 5B.7 Precision of HPLC method 126

33. 5B.8 Robustness of HPLC method 127

34. 5C.1 Saturation solubility testing of drug-cyclodextrin

complexes

130

35. 5C.2 Drug content of drug-cyclodextrin complexes 131

36. 5C.3 Multimedia dissolution of F1 132
























60. 5C.27 Percentage cumulative release of drug-cyclodextrin

complexes in 0.1 N HCl

140


complexes in phosphate buffer pH 6.8

141


complexes in water

142


complexes in OGD media

143

64. 5D.1 Incompatability testing of liquisolid excipients(Physical

observation)

166

65. 5D.2 Incompatability testing of liquisolid excipients (Assay) 166

viii

66. 5D.3 Incompatability testing of liquisolid excipients(FTIR) 167

67. 5D.4 Key formulation characteristics of liquisolid compacts of

CC with tween 20

167

68. 5D.5 Key formulation characteristics of liquisolid compacts of

CC with transcutol

168

69. 5D.6 Saturation solubility testing of liquisolid compacts 168

70. 5D.7 Drug content of liquisolid compact 169

71. 5D.8 Multimedia dissolution of Lst1 170

72. 5D.9 Multimedia dissolution of Lst2 170

73. 5D .10 Multimedia dissolution of Lst3 170







80. 5D .17 Multimedia dissolution of Lstc1 173








88. 5D.25 Multimedia dissolution of Lstc 9 175

89. 5D.26 Percentage cumulative release of liquisolid systems in

0.1N HCl

176


phosphate buffer pH 6.8

177


water

178


OGD media

179

93. 5E.1 Solubility studies of CC in modified oils , surfactants

and co surfactants

184

94. 5E.2 Stability and visual observation of SMEDDS 185-

191

95. 5E.3 Selection of SMEDDS on the basis of particle size 192

96. 5E.4 Thermodynamic stability testing of SMEDDS 192

97. 5E.5 Final selected SMEDDS 192

98. 5E.6 Drug content of SMEDDS 195

99. 5E.7 Saturation solubility testing of SMEDDS 195

100. 5E.8 Multimedia dissolution of SMEDDS 1 196



ix


104. 5E.12 Particle size distribution and polydispersity index of

SMEDDS

200

105. 5E.13 Effect of drug loading on particle size of SMEDDS. 201

106. 5F.1 Key formulation characteristics of nanoparticles with

antisolvent precipitation

204

107. 5F.2 Saturation solubility of nanoparticles with antisolvent

precipitation

204

108. 5F.3 Multimedia dissolution of nanoparticles with antisolvent

precipitation

205

109. 5F.4 Particle size distribution of nanoparticles with antisolvent

precipitation

207

110. 5F.5 Selection of nanoparticles using ion gelation technique 209

111. 5F.6 Evaluation of selected nanoparticle systems prepared by

ion-gelation technique

209

112. 5F.7 Particle size distribution of selected nanoparticle systems

prepared by ion-gelation technique

210

113. 5F.8 Saturation solubility of nanoparticles prepared by ion-

gelation technique

212

114. 5F.9 Multimedia dissolution of nanoparticles prepared by ion-

gelation technique

213

115. 5F.10 Selection of nanoparticles using nanoencpsulation

technique

215

116. 5F.11 Evaluation of selected nanoparticle systems prepared by

nanoencapsulation technique

215

117. 5F.12 Particle size distribution of selected nanoparticle systems

prepared by nanoencapsulation technique

216

118. 5F.13 Saturation solubility of nanoparticles prepared by


218

119. 5F.14 Multimedia dissolution of nanoparticles prepared by


219

120. 5G.1 Optimized formula of F16 221

121. 5G.2 Drug content and weight variation of optimized F16

formulation

221

122. 5G.3 Multimedia dissolution of optimized F16 formulation 221

123. 5G.4 Optimized formula of SMEDDS 222

124. 5G.5 Drug content and weight variation of optimized

SMEDDS formulation

222

125. 5G.6 Multimedia dissolution of optimized SMEDDS

formulation

223

126. 5H.1 Specificity for Candesartan by LCMS 226

127. 5H.2 Calibrant samples for candesartan 227

x

128. 5H.3 Curve parameter summary for Candesartan 227

129. 5H.4 Between run precision and accuracy for candesartan 228

130. 5H.5 Within run precision and accuracy for candesartan 229

131. 5H.6 Percentage extraction yield 230

132. 5H.7 Long term stock solution stability for candesartan 231

133. 5H.8 Freeze thaw stability for candesartan 232

134. 5H.9 Long term stability in matrix of candesartan 233

135. 5H.10 Variation due to change in column 234

136. 5H.11 Variation due to change in analyst 234

137. 5H.12 Variation in days for analysis (Interday variation) 235

138. 5H.13 Descriptive statistics of pharmacokinetic parameters of

candesartan(marketed formulation)

236


candesartan(F16 formulation)

236


candesartan(SMEDDS formulation)

237

141. 5I.1 Drug content of stability batches of F16 240

142. 5I.2 Dissolution of stability batches of F16 241

143. 5I.3 Drug content of stability batches of SMEDDS 243

144. 5I.4 Dissolution of stability batches of SMEDDS 244

xi

Sr.

No

Figure

No.

Name of figure Page

No.

1. 1.1 Rate limiting steps in oral absorption 1

2. 1.2 Biopharmaceutical classification system of drugs 3

3. 1.3 ADME pattern of drugs 4

4. 1.4 Factors affecting bioavailability 5

5. 1.5 Barriers for the modification of lipophilic drugs 9

6. 2.1 Chemical structure of candesartan cilexetil 24

7. 2.2 Schematic diagram of solid forms of candesartan cilexetil 36

8. 2.3 Reported DSC spectra of different forms of candesartan cilexetil

36

9. 2.4 Reported XRDspectra of different forms of candesartan cilexetil.

37

10. 2.5 Reported 13C CP/MAS spectra of Form I and Form II of candesartan cilexetil

37

11. 2.6 Reported 13C CP/MAS spectra of amorphous form of candesartan cilexetil

38

12. 2.7 Reported IR spectra of different forms of candesartan cilexetil

38

13. 4C.1 Structures of α,β and γ Cyclodextrins 54

14. 4C.2 Phase solubility profiles and classification of complexes according to Higuchi and Connors.

59

15. 4F.1 Diagram of supercritical region 89

16. 4F.2 RESS technique 90

17. 4F.3 Precipitation with compressed fluid antisolvent 91

18. 4F.4 Schematic representation of Gas antisolvent or SAS laboratory scale apparatus

91

19. 5A.1 FTIR spectra of pure drug 109

20. 5A.2 XRD spectra of pure drug 110

21. 5A.3 DSC thermogram of pure drug 110

22. 5A.4 Microscopy of pure drug 111

23. 5A.5 PH solubility profile of pure drug 111

24. 5A.6 Saturation solubility of drug 112

25. 5A.7 Multimedia dissolution of drug 113

26. 5A.8 Multimedia dissolution of marketed formulation 114

27. 5B.1 UV spectra of Pure drug in acetonitrile 115

28. 5B.2 Linearity by UV of CC in acetonitrile 115

29. 5B. 3 Linearity by UV of CC in methanol 116

30. 5B. 4 Linearity by UV of CC in water 116

31. 5B. 5 Linearity by UV of CC in 0.1 N HCl 117

32. 5B.6 Linearity by UV of CC in phosphate buffer pH 6.8 117

List of Figures

xii

33. 5B.7 Linearity by UV of CC in OGD medium 118

34. 5B.8 Chromatogram of optimized conditions of HPLC 119

35. 5B.9 Chromatogram of acid degradation(1 hour) of CC 120

36. 5B.10 Chromatogram of acid degradation (15 minutes) of CC 121

37. 5B.11 Chromatogram of degradation in base (1 hour) of CC 121

38. 5B.12 Chromatogram of degradation in base (15 minutes) of CC 122

39. 5B.13 Chromatogram of degradation by hydrolysis of CC 122

40. 5B.14 Chromatogram of placebo interference (Cyclodextrins) of CC

123

41. 5B.15 Chromatogram of placebo interference (SMEDDS excipients) of CC

123

42. 5B.16 Chromatogram of degradation by oxidation of CC 124

43. 5B.17 Linearity by HPLC method 125

44. 5C.1 Phase solubility diagram with βCD 128

45. 5C.2 Phase solubility diagram with HPβCD 128

46. 5C.3 Phase solubility diagram with γCD 129

47. 5C.4 Saturation solubility of drug-cyclodextrin complexes 131

48. 5C.5 Percentage cumulative release of drug-cyclodextrin complexes in 0.1 N HCl

144

49. 5C.6 Percentage cumulative release of drug-cyclodextrin complexes in phosphate buffer pH 6.8

144

50. 5C.7 Percentage cumulative release of drug-cyclodextrin complexes in water

145

51. 5C.8 Percentage cumulative release of drug-cyclodextrin complexes in OGD media

145

52. 5C.9 FTIR spectra of βCD 146

53. 5C.10 FTIR spectra of HPβCD 146

54. 5C.11 FTIR spectra of γCD 146

55. 5C.12 FTIR spectra of F1 147
















xiii









79. 5C.36 XRD spectra of F2 155












91. 5C.48 DSC thermogram of BCD 159

92. 5C.49 DSC thermogram of HPBCD 159

93. 5C.50 DSC thermogram of GCD 159

94. 5C.51 DSC thermogram of F2 160












106. 5D.1 Saturation solubility testing of liquisolid compacts 169

107. 5D.2 Percentage cumulative release of liquisolid system in 0.1 N HCl

176

108. 5D.3 Percentage cumulative release of liquisolid system in phosphate buffer pH 6.8

177

109. 5D.4 Percentage cumulative release of liquisolid system in water

178

110. 5D.5 Percentage cumulative release of liquisolid system in OGD media

179

111. 5D.6 Reported XRD spectra of MCC(2θ at 15.5˚ and 22˚) 180

xiv

112. 5D.7 XRD spectra of Lst1 180

113. 5D.8 XRD spectra of LStc1 180

114. 5D.9 Reported FTIR spectra of MCC 181

115. 5D.10 Reported FTIR spectra of silica 181

116. 5D.11 FTIR spectra of Lst1 182

117. 5D.12 FTIR spectra of Lstc1 182

118. 5E.1 Solubility of drug in SMEDDS excipients 184

119. 5E.2 Pseudoternary phase diagram of SMEDDS 1 193




123. 5E.6 Saturation solubility of SMEDDS 195





128. 5E.11 Intensity distribution curve of SMEDDS 1 200




132. 5F.1 Saturation solubility of nanoparticles with antisolvent precipitation

204

133. 5F.2 Multimedia dissolution of nanoparticles with antisolvent precipitation

205

134. 5F.3 X-ray diffraction spectra of nanoparticles with antisolvent precipitation

206

135. 5F.4 FTIR spectra of nanoparticles with antisolvent precipitation 206

136. 5F.5 Intensity distribution curve of nanoparticles with antisolvent precipitation

207

137. 5F.6 Intensity distribution curve of S1 210



140. 5F.9 TEM images of Nanoparticles prepared by ion gelation technique

211

141. 5F.10 Saturation solubility of nanoparticles prepared by ion gelation technique

212

142. 5F.11 Multimedia dissolution of nanoparticles prepared by ion gelation technique

213

143. 5F.12 Intensity distribution curve of NE 3 216



146. 5F.15 TEM images of nanoparticles prepared by nanoencapsulation technique

217

147. 5F.16 Saturation solubility of nanoparticles prepared by 218

xv


148. 5F.17 Multimedia dissolution of nanoparticles prepared by nanoencapsulation technique

219

149. 5G.1 Multimedia dissolution of optimized F16 formulation 222

150. 5G.2 Multimedia dissolution of optimized SMEDDS formulation

223

151. 5H.1 Representative LC-MS/MS spectra of drug (a) and internal standard(b)

225

152. 5H.2 Plasma concentration V/S time curve for marketed formulation

237

153. 5H.3 Plasma concentration V/S time curve for FD 16 formulation

237

154. 5H.4 Plasma concentration V/S time curve for SMEDDS formulation

238

155. 5H.5 Comparative data for Cmax of candesartan cilexetil 238

156. 5H.6 Comparative data for Tmax of candesartan cilexetil 238

157. 5H.7 Comparative data for AUC(0-t) of candesartan cilexetil 239

158. 5I.1 Dissolution of stability batches (real time) of F16 242

159. 5I.2 Dissolution of stability batches (accelerated) of F16 242

160. 5I.3 Dissolution of stability batches (real time) of SMEDDS 244

161. 5I.4 Dissolution of stability batches (accelerated) of SMEDDS 245

xvi

LIST OF ACRONYMS

Sr.No. Term Acronym

1 Celcius C

2 Minutes min

3 Fourier transform Infra red FTIR

4 Differential Scanning Calorimeter DSC

5 Potassium bromide KBr

6 Kilo Voltage kV

7 mili Ampere mA

8 X-ray diffraction XRD

9 Hydrochloric acid HCl

10 Normal N

11 United states pharmacopoeia USP

12 Ultra violet UV

13 Over gastric dissolution OGD

14 Molar M

15 Mili gram mg

16 Mili litre ml

17 Parts per million ppm

18 Revolutions per minute rpm

19 Nano meter nm

20 High performance liquid

chromatography

HPLC

21 Relative standard deviation RSD

22 Mili molar mM

23 Candesartan cilexitil CC

24 Micro gram µg

25 Sodium hydroxide NaOH

26 Self microemulsifying drug delivery

system

SMEDDS

27 International conference on

hormonisation

ICH

28 Beta cyclodextrin βCD

29 Hydroxypropylated beta cyclodextrin HPβCD

30 Gamma cyclodextrin √CD

31 Cyclodextrin CD

32 Supercritical Fluid SCF

33 Supercritical fluid extraction SCFE

34 Supercritical anti solvent SAS

35 Supercritical carbon dioxide SC-CO2

36 Gram gm

37 Hour hr

38 Sodium tripolyphosphate STTP

39 High density polyethylene HDPE

40 Child resistant cap CRC

41 Concentration maximum Cmax

42 Time to reach concentration maximum Tmax

xvii

43 Area under curve AUC

44 Liquid chromatographic mass

spectrophotometry mass

spectrophotometry

LC-MS/MS

45 Percent coefficient of variation %CV

46 Ethylene diamine tetra acetic acid EDTA

47 High quality control HQC

48 Low quality control LQC

49 Middle quality control MQC

50 Lower limit of quantification LLOQ

51 Upper limit of quantification ULOQ

52 Standard deviation SD

52 Certificate of analysis COA

53 Hour h

chapter no. title page no.shodhganga.inflibnet.ac.in/bitstream/10603/9096/2... · 9. 4c.1...

Documents