Supplement Materials to:

Case Studies for Practical Food Effect Assessmentsacross BCS/BDDCS Class Compounds Using In Silico, In Vitro and Preclinical In Vivo Data

METHODS

Description of Compound’s PK disposition parameters

Case Study 1: NVS732

A single intravenous (i.v.) dose of 25 mg was infused to healthy volunteers over 30 min. Blood samples were collected at predose and up to 24 h post dosing and plasma concentrations were analyzed by LC-MS/MS. The disposition parameters were determined by compartmental analysis using PKPlusTM (Simulation Plus Inc. CA, USA). A two compartment model was best fit to the concentration-time profile (Fig. s1a) and the calculated disposition parameters were used in the GastroPlus model (Table II).

Case Study 2: NVS406

A single i.v. dose of 75 mg was administered to healthy volunteers via infusion over 25 min. Blood samples were collected at predose and up to 24 h post dosing and plasma concentration was analyzed by LC-MS/MS. The human pharmacokinetic parameters were characterized by compartmental PK analysis using PKPlusTM. A three compartment PK model was found to best describe the concentration-time profile (Fig. s1b). The observed PK parameters (Table II) were then incorporated in the GastroPlus model for the prediction of oral absorption.

Case Study 3: NVS562

Human PK profiles after i.v. dosing have not been determined and therefore the disposition parameters had to be predicted from preclinical data. Preclinical PK studies with i.v. administration were conducted in mice, rats, dogs, and monkeys. The observed CL was 0.11, 0.94,0.75, and 1.5 mL/min/kg in mice, rats, dogs, and monkeys, respectively. The observed Vss was 0.49, 3.4,2.1, and 2.9L/kg in mice, rats, dogs, and monkeys, respectively. Human CL was estimated using simple allometry of the product of CL brain weight according to “the rule of exponents method” (ROE) from preclinical animal data (1) while the human Vss was estimated using simple allometry with plasma protein binding correction method. The predicted human CL and Vss are 0.015 L/h/kg and Vss 3.4 L/kg, respectively. The normalized concentration-time curve at 1 mg/kg in humans was then projected based on the concentration-time curves from preclinical species using Wajima et al. method (2). A two compartmental PK model was found to best describe the projected human concentration-time curve and the calculated PK parameters were used in the model (Table II).

Case Study 4: NVS701

Human PK profiles after i.v. doses have not been determined and therefore the disposition parameters were predicted from preclinical data. The observed CL was 5.0, 4.3,8.8, and 11.0 ml/min/kg in mice, rats, dogs, and monkeys, respectively. The observed Vss was 0.52, 7.9,2.5, and 0.67L/kg in mice, rats, dogs, and monkeys, respectively. Human CL was estimated using fraction unbound (fu) intercept correction method between rat and human(1) according to ROE while the human Vss was estimated using Oie-Tozer method (3-4). The predicted human CL and Vss are 0.10 L/h/kg and 2.1 L/kg, respectively.Further, the human concentration-time curve was projected using Wajima et al. method (2). A two compartmental PK model best described the projected human concentration-time curve and the calculated PK parameters were used in the model (Table II).

Case Study 5: NVS001

A single i.v. dose of 20 mg was administered to healthy volunteers via infusion over 15 min. Blood samples will be collected at pre-dose and up to 12 h post dosing and plasma concentration was analyzed by LC-MS/MS. The PK parameters of was characterized by compartmental PK analysis using PKPlusTM. A three compartment PK model was found to best describe the concentration-time profile (Fig. s1c) and observed PK parameters are listed (Table II).

Case Study 6: NVS169 and NVS113

Human PK profiles after i.v. doses have not been determined for both compounds and therefore the disposition parameters were predicted from preclinical data. Preclinical PK studies with i.v. doses were conducted in rats and dogs, NVS169 showed moderate CL (34 mL/min/kg) and moderate Vss (2.1 L/kg) in rats while exhibited low CL (4.0 mL/min/kg) and low Vss (0.3 L/kg) in dogs. Using a single species scaling method from rats as a conservative estimation, human CL and Vss were predicted to be 0.38 L/h/kg and 1.3 L/kg, respectively. A two compartment model was assumed for NVS169 to describe human PK profiles and the calculated PK parameters were used in the model (Table II). For NVS113, Preclinical PK studies with i.v. doses were conducted in rats, dogs, and monkeys. The observed CL was 1.4, 0.56 and 0.57 mL/min/kg in rats, dogs, and monkeys, respectively. The observed Vss was 0.59, 0.28,and 0.59L/kg in rats, dogs, and monkeys, respectively. The human CL was estimated using allometric scaling according to ROE (1) while the human Vss was estimated using Oie-Tozer method (3, 4) from the above three preclinical species. The predicted human CL and Vss are 0.018 L/h/kg and Vss 0.4 L/kg, respectively. Similar, the human concentration-time curve was then projected using Wajima et al. method (2). A two compartmental PK model best described the projected human concentration-time curve and the calculated PK parameters were used in the model (Table II).

SUPPLEMENTARY TABLES

SupplementaryTable S1: Relative transporter expressions used in GastroPlus Simulation.

Transporters / Duodenum / Jejunum 1 / Jejunum 2 / Ileum 1 / Ileum 2 / Ileum 3 / Caecum/Colon
P-gpa / 0.538 / 0.645 / 0.723 / 0.77 / 0.838 / 0.908 / 1
OATP2B1b / 0.33 / 0.33 / 0.33 / 1 / 1 / 1 / 0.33

a From (5)

b From (6)

Supplementary Table S2: Simulated AUC values of NVS562 with mean precipitation time (MPT) generated from in vitro or in vivo data

Formulation / Fed state / Mean Precipitation Time (s) / AUC0-last (ng∙h/mL)
Calculated from in vitro data / Optimized from clinical in vivo data / Predicted usingin vitro MPT / Predicted using in vivo MPT / Observed
SEDDS / Fasted / 3564 / 6000 / 13670 / 14230 / 14100
Fed / 3478 / 6000 / 12810 / 13210 / 14260
Tablet / Fasted / 2210 / 3500 / 5728 / 6909 / 7480
Fed / 3231 / 6000 / 9722 / 10320 / 11900

LEGENDS OF SUPPLEMENTARY FIGURES

Supplementary Figure S1. Mean observed plasma concentration (solid circle) and fitted

plasma concentration time curve (black line) after a single i.v. infusion dose of (a) 25 mg of NVS732, (b) 75 mg of NVS406, or (c) 20 mg of NVS001 in healthy volunteers.

Supplementary Figure S2. Mean observed food effect after oral administration of 3 mg/kg of NVS001 in fasted (black) andnon-fasted primates (gray).

SUPPLEMENTARY FIGURES

Figure S1.

Figure S2.

REFERENCES FOR SUPPLEMENTAL DOCUMENTS

1.Mahmood I. Prediction of human drug clearance from animal data: application of the rule of exponents and 'fu Corrected Intercept Method' (FCIM). J Pharm Sci. 2006;95(8):1810-21. DOI: 10.1002/jps.20650.

2.Wajima T, Yano Y, Fukumura K, Oguma T. Prediction of human pharmacokinetic profile in animal scale up based on normalizing time course profiles. J Pharm Sci. 2004;93(7):1890-900.

3.Oie S, Tozer TN. Effect of Altered Plasma-Protein Binding on Apparent Volume of Distribution. J Pharm Sci. 1979;68(9):1203-5.

4.Lombardo F, Obach RS, Shalaeva MY, Gao F. Prediction of volume of distribution values in humans for neutral and basic drugs using physicochemical measurements and plasma protein binding data. J Med Chem. 2002;45(13):2867-76. DOI: 10.1021/Jm0200409.

5.Mouly S, Paine MF. P-glycoprotein increases from proximal to distal regions of human small intestine. Pharm Res. 2003;20(10):1595-9.

6.Meier Y, Eloranta JJ, Darimont J, Ismair MG, Hiller C, Fried M, et al. Regional distribution of solute carrier mRNA expression along the human intestinal tract. Drug MetabDispos. 2007;35(4):590-4. DOI: 10.1124/dmd.106.013342.