Modulation of Pharmacokinetic and Cytotoxicity Profile of Imatinib Base by Employing Optimized

Modulation of Pharmacokinetic and Cytotoxicity Profile of Imatinib Base by Employing Optimized Nanostructured Lipid Carriers

BikiGuptaa, Bijay Kumar Poudela, Tuan HiepTrana, Roshan Pradhana, Hyuk-Jun Choa, Jee-HeonJeonga, Beom Soo Shinb, Han-GonChoic, Chul Soon Yonga*, Jong Oh Kima*

aCollege of Pharmacy, Yeungnam University, 214-1, Dae-Dong,Gyeongsan 712-749, South Korea

bCollege of Pharmacy, Catholic University of Daegu, Gyeongsan, 712-702, South Korea

c College of Pharmacy, Institute of Pharmaceutical Science and Technology, Hanyang University, 55, Hanyangdaehak-ro, Sangnok-gu,Ansan 426-791, South Korea

*Corresponding author: Prof. Jong Oh Kim, Ph.D.,

Tel: +82-53-810-2813, Fax: +82-53-810-4654,

E-mail:

*Co-corresponding author: Prof. Chul Soon Yong, Ph.D.

Tel: +82-53-810-2812, Fax: +82-53-810-4654,

E-mail:

Table S1. Predefined level of variables for NLC test formulations

Variable / Predefined level
Oil to total lipid ratio, O/L / 0.50
Organic to aqueous phase ratio, O/A / 0.05
Amount of Tween 20, Tw20 a / 1.5% w/v
Amount of Lecithin, Lecb / 2% w/w
Homogenization time, HT / 10 min
Sonication time, ST / 8 min
Sonication amplitude, SA / 70%

a relative to aqueous phase, b relative to total lipid

Table S2. Blank NLC formulations for preliminary testing

Formulation ID / Solid lipid–Liquid lipid pair / Particle size (nm)a / PDIa
BF-1b / CmA – LfW / - / -
BF-2 / CmA – LbM / 180.17 ± 4.26 / 0.198 ± 0.028
BF-3 / CmA – LbF / 178.67 ± 3.39 / 0.162 ± 0.031
BF-4b / CmA – LbW / - / -
BF-5b / PcA – LfW / - / -
BF-6 / PcA – LbM / 161.33 ± 2.78 / 0.169 ± 0.036
BF-7 / PcA – LbF / 169.60 ± 1.71 / 0.177 ± 0.018
BF-8b / PcA – LbW / - / -
BF-9b / GlC – LfW / - / -
BF-10 / GlC – LbM / 311.63 ± 3.46 / 0.324 ± 0.062
BF-11 / GlC – LbF / 292.90 ± 5.12 / 0.316 ± 0.021
BF-12b / GlC – LbW / - / -
BF-13b / SpA – LfW / - / -
BF-14 / SpA – LbM / 443.40 ± 12.41 / 0.464 ± 0.060
BF-15 / SpA – LbF / 488.90 ± 21.51 / 0.471 ± 0.021
BF-16b / SpA – LbW / - / -

aeach value represented as mean ± SD (n=3); b formulations not fairly stable upon preparation. [CmA: Compritol 888 ATO; PcA: Precirol ATO 5; GlC: Gelucire 44/14; SpA: Sappocire AP; LfW: Labrafaclipophile WL 1349; LbM: Labrafil M 1944 CS; LbF: Labrafil M 2125 CS; LbW: Labrafil WL 2609 BS]

Table S3. Imatinib-loaded NLC formulations for preliminary testing

Formulation ID / Solid lipid / liquid lipid pair / Size (nm)a / PDIa / EE (%w/w) a
F-1 / CmA / LbM / 181.27 ± 0.61 / 0.174 ± 0.017 / 89.14 ± 0.03
F-2 / CmA / LbF / 182.00 ± 3.85 / 0.172 ± 0.014 / 88.26 ± 0.06
F-3 / PcA / LbM / 166.00 ± 0.20 / 0.187 ± 0.009 / 95.38 ± 0.13
F-4 / PcA / LbF / 171.80 ± 0.44 / 0.195 ± 0.006 / 92.41 ± 0.09

aeach value represented as mean ± SD (n=3) [CmA: Compritol 888 ATO; PcA: Precirol ATO 5; LbM: Labrafil M 1944 CS; LbF: Labrafil M 2125 CS]

Table S4. Plackett-Burman Design Matrix with Observed Responses

Run / x1 / x2 / x3(%w/w) / x4(%w/v) / x5 / x6(min) / x7(min) / x8(%) / x9 / x10 / x11 / y1(%w/w) / y2(%w/w) / y3(nm) / PDI
1 / 0.3 / 0.06 / 6 / 1 / 0.06 / 15 / 10 / 60 / -1 / -1 / 1 / 5.89 / 98.14 / 195.57 / 0.215
2 / 0.5 / 0.02 / 6 / 3 / 0.06 / 9 / 6 / 60 / 1 / -1 / 1 / 5.70 / 95.05 / 112.13 / 0.292
3 / 0.5 / 0.06 / 6 / 1 / 0.04 / 9 / 10 / 60 / 1 / 1 / -1 / 3.90 / 97.46 / 186.33 / 0.186
4 / 0.3 / 0.02 / 2 / 3 / 0.04 / 15 / 10 / 60 / 1 / 1 / 1 / 3.70 / 92.39 / 105.97 / 0.142
5 / 0.4 / 0.04 / 4 / 2 / 0.05 / 12 / 8 / 70 / 0 / 0 / 0 / 4.82 / 96.32 / 67.56 / 0.229
6 / 0.4 / 0.04 / 4 / 2 / 0.05 / 12 / 8 / 70 / 0 / 0 / 0 / 4.82 / 96.34 / 67.07 / 0.222
7 / 0.3 / 0.06 / 2 / 3 / 0.06 / 9 / 10 / 80 / 1 / -1 / -1 / 5.88 / 98.05 / 64.14 / 0.185
8 / 0.5 / 0.02 / 6 / 3 / 0.04 / 15 / 10 / 80 / -1 / -1 / -1 / 3.71 / 92.77 / 133.33 / 0.146
9 / 0.5 / 0.06 / 2 / 1 / 0.04 / 15 / 6 / 80 / 1 / -1 / 1 / 3.83 / 95.85 / 198.50 / 0.219
10 / 0.5 / 0.02 / 2 / 1 / 0.06 / 9 / 10 / 80 / -1 / 1 / 1 / 5.47 / 91.16 / 59.08 / 0.173
11 / 0.3 / 0.06 / 6 / 3 / 0.04 / 9 / 6 / 80 / -1 / 1 / 1 / 3.87 / 96.78 / 82.10 / 0.227
12 / 0.3 / 0.02 / 2 / 1 / 0.04 / 9 / 6 / 60 / -1 / -1 / -1 / 3.55 / 88.73 / 88.89 / 0.286
13 / 0.3 / 0.02 / 6 / 1 / 0.06 / 15 / 6 / 80 / 1 / 1 / -1 / 5.59 / 93.16 / 74.88 / 0.229
14 / 0.5 / 0.06 / 2 / 3 / 0.06 / 15 / 6 / 60 / -1 / 1 / -1 / 5.87 / 97.82 / 86.90 / 0.281
15 / 0.4 / 0.04 / 4 / 2 / 0.05 / 12 / 8 / 70 / 0 / 0 / 0 / 4.80 / 96.05 / 61.37 / 0.230

[x1: oil to total lipid ratio; x2: organic to aqueous phase ratio; x3: amount of lecithin; x4: amount of Tween® 20; x5: drug to total lipid ratio; x6: homogenization time; x7: sonication time; x8: sonication amplitude; x9: dummy1; x10: dummy2; x11: dummy3; y1: drug loading; y2: encapsulation efficiency; y3: particle size]

Table S5. Central Composite Design Matrix with Observed Responses

Run / X1 / X2 / X3
(%w/w) / X4
(%w/v) / Y1
(%w/w) / Y2
(%w/w) / Y3
(%) / Y4
(nm) / PDI
1 / 0.040 / 0.05 / 7.0 / 2.00 / 4.35 / 96.77 / 81.62 / 70.73 / 0.278
2 / 0.055 / 0.04 / 2.5 / 1.25 / 3.59 / 96.09 / 54.94 / 157.40 / 0.218
3 / 0.025 / 0.06 / 2.5 / 2.75 / 5.34 / 95.21 / 87.38 / 67.66 / 0.208
4 / 0.025 / 0.04 / 2.5 / 2.75 / 3.49 / 93.39 / 83.10 / 67.30 / 0.160
5 / 0.040 / 0.03 / 4.0 / 2.00 / 2.61 / 94.35 / 86.78 / 60.62 / 0.220
6 / 0.040 / 0.05 / 4.0 / 2.00 / 4.44 / 96.23 / 76.30 / 60.28 / 0.240
7 / 0.010 / 0.05 / 4.0 / 2.00 / 4.12 / 88.98 / 99.76 / 71.67 / 0.130
8 / 0.040 / 0.07 / 4.0 / 2.00 / 6.30 / 97.51 / 79.26 / 67.47 / 0.198
9 / 0.040 / 0.05 / 4.0 / 2.00 / 4.45 / 96.44 / 89.57 / 68.18 / 0.225
10 / 0.025 / 0.06 / 5.5 / 1.25 / 5.23 / 95.65 / 74.18 / 68.76 / 0.210
11 / 0.040 / 0.05 / 4.0 / 2.00 / 4.47 / 96.87 / 86.42 / 67.96 / 0.221
12 / 0.025 / 0.04 / 2.5 / 1.25 / 3.53 / 94.27 / 77.18 / 65.42 / 0.199
13 / 0.055 / 0.04 / 2.5 / 2.75 / 3.63 / 97.15 / 85.21 / 66.42 / 0.215
14 / 0.025 / 0.06 / 2.5 / 1.25 / 5.38 / 95.98 / 84.10 / 63.93 / 0.205
15 / 0.040 / 0.05 / 4.0 / 2.00 / 4.49 / 97.36 / 82.06 / 67.23 / 0.207
16 / 0.040 / 0.05 / 1.0 / 2.00 / 4.58 / 96.69 / 86.82 / 64.96 / 0.181
17 / 0.025 / 0.04 / 5.5 / 1.25 / 3.40 / 93.39 / 68.32 / 70.22 / 0.217
18 / 0.025 / 0.06 / 5.5 / 2.75 / 5.25 / 95.93 / 88.28 / 74.17 / 0.165
19 / 0.040 / 0.05 / 4.0 / 2.00 / 4.46 / 96.75 / 84.53 / 66.79 / 0.193
20 / 0.040 / 0.05 / 4.0 / 3.50 / 4.50 / 97.45 / 94.33 / 72.85 / 0.133
21 / 0.055 / 0.06 / 5.5 / 2.75 / 5.40 / 98.70 / 90.09 / 69.24 / 0.212
22 / 0.055 / 0.06 / 2.5 / 1.25 / 5.50 / 98.00 / 70.11 / 152.87 / 0.179
23 / 0.055 / 0.04 / 5.5 / 2.75 / 3.54 / 97.16 / 88.46 / 70.17 / 0.223
24 / 0.055 / 0.04 / 5.5 / 1.25 / 3.52 / 96.69 / 58.36 / 155.80 / 0.204
25 / 0.070 / 0.05 / 4.0 / 2.00 / 4.51 / 97.73 / 76.09 / 132.40 / 0.231
26 / 0.025 / 0.04 / 5.5 / 2.75 / 3.44 / 94.41 / 97.71 / 72.66 / 0.174
27 / 0.040 / 0.05 / 4.0 / 2.00 / 4.47 / 96.86 / 80.77 / 66.35 / 0.208
28 / 0.055 / 0.06 / 5.5 / 1.25 / 5.34 / 97.59 / 64.74 / 150.87 / 0.207
29 / 0.040 / 0.05 / 4.0 / 0.50 / 4.45 / 96.33 / 82.49 / 178.57 / 0.248
30 / 0.055 / 0.06 / 2.5 / 2.75 / 5.49 / 97.79 / 78.37 / 67.48 / 0.168

[X1: organic to aqueous phase ratio; X2: drug to total lipid ratio; X3: amount of lecithin; X4: amount of Tween® 20; Y1: drug loading; Y2: encapsulation efficiency; Y3: drug release at 36 h; Y4: particle size]

Table S6. Correlation Coefficients for the Response Surface Reduced Models

Response / R2 / Adjusted R2 / Predicted R2 / Adequate precision
Y1 / 0.9990 / 0.9988 / 0.9976 / 315.496
Y2 / 0.9004 / 0.8889 / 0.7656 / 34.430
Y3 / 0.5731 / 0.5415 / 0.4359 / 12.803
Y4 / 0.9848 / 0.9816 / 0.9532 / 49.227

Table S7. Fit Summary for Response Variables

Response / Source / Sequential p-value / Lack of Fit p-value / Remarks
Y1 / Linear / < 0.0001 / 0.0132
SFI / 0.9126 / 0.0078
Quadratic / 0.0002 / 0.0889 / Suggested
Cubic / 0.1387 / 0.1425
Y2 / Linear / < 0.0001 / 0.0123
2FI / 0.9647 / 0.0067
Quadratic / 0.0003 / 0.0756 / Suggested
Cubic / 0.1698 / 0.0978
Y3 / Linear / 0.0002 / 0.1262 / Suggested
2FI / 0.2228 / 0.1480
Quadratic / 0.8476 / 0.0994
Cubic / 0.7485 / 0.0234
Y4 / Linear / 0.0003 / < 0.0001
2FI / 0.0295 / < 0.0001
Quadratic / < 0.0001 / 0.0501 / Suggested
Cubic / 0.0060 / 0.9704

Table S8. Factor Coefficient Estimates and Corresponding Standard Error Values

Factor / Y1 / Y2 / Y3 / Y4
Coeff. estimate / Std. error / Coeff. estimate / Std. error / Coeff. estimate / Std. error / Coeff. estimate / Std. error
Intercept / 4.47 / 6.751x10-3 / 96.76 / 0.15 / 81.24 / 1.28 / 66.08 / 1.41
X1 / 0.072 / 5.846x10-3 / 1.60 / 0.13 / - 4.89 / 1.43 / 19.23 / 0.99
X2 / 0.92 / 5.846x10-3 / 0.78 / 0.13 / - / - / - / -
X3 / - 0.053 / 5.846x10-3 / - / - / - / - / - / -
X4 / - / - / - / - / 7.10 / 1.43 / -22.57 / 0.99
X1X4 / - / - / - / - / - / - / -22.32 / 1.22
X12 / - 0.036 / 5.337x10-3 / -0.79 / 0.12 / - / - / 9.00 / 0.91
X42 / - / - / - / - / - / - / 14.92 / 0.91

Note: The insignificant factors were excluded from the model to obtain a better fit.