The AAPS Journal

, Volume 15, Issue 3, pp 763–774 | Cite as

Assessment of Juvenile Pigs to Serve as Human Pediatric Surrogates for Preclinical Formulation Pharmacokinetic Testing

  • Wyatt J. Roth
  • Candice B. Kissinger
  • Robyn R. McCain
  • Bruce R. Cooper
  • Jeremy N. Marchant-Forde
  • Rachel C. Vreeman
  • Sophia Hannou
  • Gregory T. KnippEmail author
Research Article Theme: Challenges and Opportunities in Pediatric Drug Development


Pediatric drug development is hampered by biological, clinical, and formulation challenges associated with age-based populations. A primary cause for this lack of development is the inability to accurately predict ontogenic changes that affect pharmacokinetics (PK) in children using traditional preclinical animal models. In response to this issue, our laboratory has conducted a proof-of-concept study to investigate the potential utility of juvenile pigs to serve as surrogates for children during preclinical PK testing of selected rifampin dosage forms. Pigs were surgically modified with jugular vein catheters that were externalized in the dorsal scapular region and connected to an automated blood sampling system (PigTurn-Culex-L). Commercially available rifampin capsules were administered to both 20 and 40 kg pigs to determine relevant PK parameters. Orally disintegrating tablet formulations of rifampin were also developed and administered to 20 kg pigs. Plasma samples were prepared from whole blood by centrifugation and analyzed for rifampin content by liquid chromatography–tandem mass spectrometry. Porcine PK parameters were determined from the resultant plasma–concentration time profiles and contrasted with published rifampin PK data in human adults and children. Results indicated significant similarities in dose-normalized absorption and elimination parameters between pigs and humans. Moreover, ontogenic changes observed in porcine PK parameters were consistent with ontogenic changes reported for human PK. These results demonstrate the potential utility of the juvenile porcine model for predicting human pediatric PK for rifampin. Furthermore, utilization of juvenile pigs during formulation testing may provide an alternative approach to expedite reformulation efforts during pediatric drug development.


ADME pediatric pharmacokinetics porcine rifampin 



The authors would like to acknowledge the financial support received from the Indiana Clinical and Translational Sciences Institute (NIH Grant # RR025761, Dr. Knipp and Dr. Vreeman), the Dane O. Kildsig Center for Pharmaceutical Processing and Research, and the 2011–2012 Lilly Endowment Graduate Research Fellowship for Mr. Wyatt Roth. The authors would like to acknowledge that the porcine studies were conducted in the Purdue Translational Pharmacology (PTP) Core Facility of the Clinical Translational Sciences Award (Core Pilot Funding NIH Grant # RR025761). In addition, we would also like to thank Bioanalytical Systems, Inc. and Dr. Jeremy Marchant-Forde for providing access to the PigTurn-Culex-L® units. We would also like to acknowledge the assistance of Dr. Lee Matthews from the Purdue Office of the Vice President for Research for serving as the staff veterinarian for these studies. Finally, we thank Drs. Peter Kissinger, Carmen Popescu, and Rodolfo Pinal for their helpful suggestions and assistance on the project.


  1. 1.
    Shirkey H. Therapeutic orphans. J Pediatr. 1968;72:119–20.PubMedCrossRefGoogle Scholar
  2. 2.
    Milne CP, Bruss JB. The economics of pediatric formulation development for off-patent drugs. Clin Ther. 2008;30:2133–45.PubMedCrossRefGoogle Scholar
  3. 3.
    Hoppu K, Anabwani G, Garcia-Bournissen F, Gazarian M, Kearns GL, Nakamura H, et al. The status of paediatric medicines initiatives around the world—what has happened and what has not? Eur J Clin Pharmacol. 2012;68:1–10.PubMedCrossRefGoogle Scholar
  4. 4.
    Rose K. Challenges in pediatric drug development: a pharmaceutical industry perspective. Paediatr Drugs. 2009;11:57–9.PubMedCrossRefGoogle Scholar
  5. 5.
    Gazarian M. Delivering better medicines to children: need for better integration between the science, the policy, and the practice. Paediatr Drugs. 2009;11:41–4.PubMedCrossRefGoogle Scholar
  6. 6.
    Schirm E, Tobi H, de Vries TW, Choonara I, De Jong-van den Berg LT. Lack of appropriate formulations of medicines for children in the community. Acta Paediatr. 2003;92:1486–9.PubMedCrossRefGoogle Scholar
  7. 7.
    Nunn T, Williams J. Formulation of medicines for children. Br J Clin Pharmacol. 2005;59:674–6.PubMedCrossRefGoogle Scholar
  8. 8.
    Walters S. Report to WHO concerning international guidelines for paediatric medicines. Geneva: World Health Organization; 2010. Accessed 30 Aug 2012
  9. 9.
    Lam MS. Extemporaneous compounding of oral liquid dosage formulations and alternative drug delivery methods for anticancer drugs. Pharmacotherapy. 2011;31:164–92.PubMedCrossRefGoogle Scholar
  10. 10.
    Blake MJ, Abdel-Rahman SM, Jacobs RF, Lowery NK, Sterling TR, Kearns GL. Pharmacokinetics of rifapentine in children. Pediatr Infect Dis J. 2006;25:405–9.PubMedCrossRefGoogle Scholar
  11. 11.
    Notterman DA, Nardi M, Saslow JG. Effect of dose formulation on isoniazid absorption in two young children. Pediatrics. 1986;77:850–2.PubMedGoogle Scholar
  12. 12.
    Giacoia GP, Taylor-Zapata P, Mattison D. Eunice Kennedy Shriver National Institute of Child Health and Human Development Pediatric Formulation Initiative: selected reports from working groups. Clin Ther. 2008;30:2097–101.PubMedCrossRefGoogle Scholar
  13. 13.
    De Cock RF, Piana C, Krekels EH, Danhof M, Allegaert K, Knibbe CA. The role of population PK-PD modelling in paediatric clinical research. Eur J Clin Pharmacol. 2011;67:5–16.PubMedCrossRefGoogle Scholar
  14. 14.
    Yu LX, Ellison CD, Hussain AS. Predicting human oral bioavailability using in silico models In: Krishna R, editor. Applications of pharmacokinetic principles in drug development. New York: Kluwer Academic/Plenum; 2004. p. 53–72.CrossRefGoogle Scholar
  15. 15.
    Grass GM, Sinko PJ. Physiologically-based pharmacokinetic simulation modelling. Adv Drug Deliv Rev. 2002;54:433–51.PubMedCrossRefGoogle Scholar
  16. 16.
    Swindle MM, Makin A, Herron AJ, Clubb FJ, Frazier KS. Swine as models in biomedical research and toxicology testing. Veterinary Pathology. 2012;49:344–56.PubMedCrossRefGoogle Scholar
  17. 17.
    DeSesso JM, Williams AL. Contrasting the gastrointestinal tracts of mammals: factors that influence absorption. Ann Rep Med Chem. 2008;43:353–71.CrossRefGoogle Scholar
  18. 18.
    Svendsen O. The minipig in toxicology. Exp Toxicol Pathol. 2006;57:335–9.PubMedCrossRefGoogle Scholar
  19. 19.
    FDA. Guidance for industry nonclinical safety evaluation of pediatric drug products. Silver Spring: Food and Drug Administration; 2006. Accessed 30 Aug 2012.
  20. 20.
    EMA. Guideline on the need for non-clinical testing in juvenile animals of pharmaceuticals for paediatric indications. London: European Medicines Agency; 2005. Accessed 30 Aug 2012.
  21. 21.
    Baldrick P. Juvenile animal testing in drug development—is it useful? Reg Tox Pharmacol. 2010;57:291–9.CrossRefGoogle Scholar
  22. 22.
    Bode G, Clausing P, Gervais F, Loegsted J, Luft J, Nogues V, et al. The utility of the minipig as an animal model in regulatory toxicology. J Pharmacol Toxicol Methods. 2010;62:196–220.PubMedCrossRefGoogle Scholar
  23. 23.
    Forster R, Ancian P, Fredholm M, Simianer H, Whitelaw B. The minipig as a platform for new technologies in toxicology. J Pharmacol Toxicol Methods. 2010;62:227–35.PubMedCrossRefGoogle Scholar
  24. 24.
    Forster R, Bode G, Ellegaard L, van der Laan JW. The RETHINK project—minipigs as models for the toxicity testing of new medicines and chemicals: an impact assessment. J Pharmacol Toxicol Methods. 2010;62:158–9.PubMedCrossRefGoogle Scholar
  25. 25.
    Forster R, Bode G, Ellegaard L, van der Laan JW. The RETHINK project on minipigs in the toxicity testing of new medicines and chemicals: conclusions and recommendations. J Pharmacol Toxicol Methods. 2010;62:236–42.PubMedCrossRefGoogle Scholar
  26. 26.
    Kararli TT. Comparison of the gastrointestinal anatomy, physiology, and biochemistry of humans and commonly used laboratory-animals. Biopharm Drug Dispos. 1995;16:351–80.PubMedCrossRefGoogle Scholar
  27. 27.
    van der Laan JW, Brightwell J, McAnulty P, Ratky J, Stark C, Project R. Regulatory acceptability of the minipig in the development of pharmaceuticals, chemicals and other products. J Pharmacol Toxicol Methods. 2010;62:184–95.PubMedCrossRefGoogle Scholar
  28. 28.
    Skaanild MT, Friis C. Characterization of the P450 system in Gottingen minipigs. Pharmacol Toxicol. 1997;80:28–33.PubMedCrossRefGoogle Scholar
  29. 29.
    Skaanild MT, Friis C. Cytochrome P450 sex differences in minipigs and conventional pigs. Pharmacol Toxicol. 1999;85:174–80.PubMedCrossRefGoogle Scholar
  30. 30.
    Skaanild MT, Friis C. Porcine CYP2A polymorphisms and activity. Basic Clin Pharmacol Toxicol. 2005;97:115–21.PubMedCrossRefGoogle Scholar
  31. 31.
    Skaanild MT. Porcine cytochrome P450 and metabolism. Current Pharm Design. 2006;12:1421–7.CrossRefGoogle Scholar
  32. 32.
    Fink-Gremmels J. Implications of hepatic cytochrome P450-related biotransformation processes in veterinary sciences. Eur J Pharmacol. 2008;585:502–9.PubMedCrossRefGoogle Scholar
  33. 33.
    Goh LB, Spears KJ, Yao DG, Ayrton A, Morgan P, Wolf CR, et al. Endogenous drug transporters in in vitro and in vivo models for the prediction of drug disposition in man. Biochem Pharmacol. 2002;64:1569–78.PubMedCrossRefGoogle Scholar
  34. 34.
    WHO. 3rd WHO model list of essential medicines for children. Geneva: World Health Organization; 2011. Accessed 2 Jun 2012.
  35. 35.
    US Pharmacopeia. Rifampin capsules. Rockville: US Pharmacopeia; 2012. 1, 2012. Accessed 23 Jan 2012.
  36. 36.
    Liu JF, Sun J, Zhang W, Gao K, He ZG. HPLC determination of rifampicin and related compounds in pharmaceuticals using monolithic column. J Pharm Biomed Analysis. 2008;46:405–9.CrossRefGoogle Scholar
  37. 37.
    Marchant-Forde J, Matthews D, Poletto R, McCain R, Mann D, DeGraw R, et al. Plasma cortisol and noradrenalin concentrations in pigs: automated sampling of freely moving pigs housed in the PigTurn(R) versus manually sampled and restrained pigs Animal Welfare. 2012;21:197–205.CrossRefGoogle Scholar
  38. 38.
    Naidong W, Shou WZ, Addison T, Maleki S, Jiang X. Liquid chromatography/tandem mass spectrometric bioanalysis using normal-phase columns with aqueous/organic mobile phases—a novel approach of eliminating evaporation and reconstitution steps in 96-well SPE. Rapid Commun Mass Spectrom. 2002;16:1965–75.PubMedCrossRefGoogle Scholar
  39. 39.
    Hartkoorn RC, Khoo S, Back DJ, Tjia JF, Waitt CJ, Chaponda M, et al. A rapid and sensitive HPLC-MS method for the detection of plasma and cellular rifampicin. J Chromatogr B Analyt Technol Biomed Life Sci. 2007;857:76–82.PubMedCrossRefGoogle Scholar
  40. 40.
    US Pharmacopeia. Rifampin capsules. Rockville: US Pharmacopeia; 2012 Accessed 10 Sep 2012.
  41. 41.
    US Pharmacopeia. Disintegration. Rockville: US Pharmacopeia; 2012 Accessed 10 Sep 2012
  42. 42.
    Gal JY, Fovet Y, Adib-Yadzi M. About a synthetic saliva for in vitro studies. Talanta. 2001;53:1103–15.PubMedCrossRefGoogle Scholar
  43. 43.
    Acocella G, Pagani V, Marchetti M, Baroni GC, Nicolis FB. Kinetic studies on rifampicin. I. Serum concentration analysis in subjects treated with different oral doses over a period of two weeks. Chemotherapy. 1971;16:356–70.PubMedCrossRefGoogle Scholar
  44. 44.
    Acocella G. Pharmacokinetics and metabolism of rifampin in humans. Rev Infect Dis. 1983;5:S428–32.PubMedCrossRefGoogle Scholar
  45. 45.
    Acocella G. Clinical pharmacokinetics of rifampicin. Clin Pharmacokinet. 1978;3:108–27.PubMedCrossRefGoogle Scholar
  46. 46.
    Schaaf HS, Willemse M, Cilliers K, Labadarios D, Maritz JS, Hussey GD, et al. Rifampin pharmacokinetics in children, with and without human immunodeficiency virus infection, hospitalized for the management of severe forms of tuberculosis. BMC Med. 2009;7:19.PubMedCrossRefGoogle Scholar
  47. 47.
    Hussels H, Kroening U, Magdorf K. Ethambutol and rifampicin serum levels in children: second report on the combined administration of ethambutol and rifampicin. Pneumonogie. 1973;149:31–8.CrossRefGoogle Scholar
  48. 48.
    McCracken G, Ginsburg C, Zweighaft T, Clahsen J. Pharmacokinetics of rifampin in infants and children: relevance to prophylaxis against Haemophilus influenzae type B disease. Pediatrics. 1980;66:17–21.PubMedGoogle Scholar
  49. 49.
    Seth V, Beotra A, Bagga A, Seth S. Drug therapy in malnutrition. Indian J Pediatr. 1992;29:1341–6.Google Scholar
  50. 50.
    Seth V, Beotra A, Seth S, Semwal O, Kabra S, Jain Y. Serum concentrations of rifampicin and isoniazid in tuberculosis. Indian J Pediatr. 1993;30:1091–8.Google Scholar
  51. 51.
    Thee S, Detjen A, Wahn U, Magdorf K. Rifampicin serum levels in childhood tuberculosis. Int J Tuberc Lung Dis. 2009;13:1106–11.PubMedGoogle Scholar
  52. 52.
    Donald PR, Maritz JS, Diacon AH. The pharmacokinetics and pharmacodynamics of rifampicin in adults and children in relation to the dosage recommended for children. Tuberculosis. 2011;91:196–207.PubMedCrossRefGoogle Scholar
  53. 53.
    Bruzzese T, Rimaroli C, Bonabello A, Mozzi G, Ajay S, Cooverj ND. Pharmacokinetics and tissue distribution of rifametane, a new 3-azinomethyl-rifamycin derivative, in several animal species. Arzneimittelforschung. 2000;50:60–71.PubMedGoogle Scholar
  54. 54.
    Pallanza R, Arioli V, Furesz S, Bolzoni G. Rifampicin: a new rifamycin. II. Laboratory studies on the antituberculous activity and preliminary clinical observations. Arzneimittelforschung. 1967;17:529–34.PubMedGoogle Scholar
  55. 55.
    Venturini AP. Pharmacokinetics of L/105, a new rifamycin, in rats and dogs, after oral administration. Chemotherapy. 1983;29:1–3.PubMedCrossRefGoogle Scholar
  56. 56.
    Agrawal S, Panchagnula R. Implication of biopharmaceutics and pharmacokinetics of rifampicin in variable bioavailability from solid oral dosage forms. Biopharm Drug Dispos. 2005;26:321–34.PubMedCrossRefGoogle Scholar
  57. 57.
    Finkel JM, Pittillo RF, Mellett LB. Fluorometric and microbiological assays for rifampicin and the determination of serum levels in the dog. Chemotherapy. 1971;16:380–8.PubMedCrossRefGoogle Scholar
  58. 58.
    Stetter MD, Peloquin CA. Isoniazid and rifampin serum levels in a Colobus monkey (Colobus guereza caudatus) infected with Mycobacterium bovis. J Zoo Wildlife Med. 1995;26:152–4.Google Scholar
  59. 59.
    Ohtsuka T, Yoshikawa T, Kozakai K, Tsuneto Y, Uno Y, Utoh M, et al. Alprazolam as an in vivo probe for studying induction of CYP3A in cynomolgus monkeys. Drug Metab Dispos. 2010;38:1806–13.PubMedCrossRefGoogle Scholar
  60. 60.
    Henwood SQ, de Villiers MM, Liebenberg W, Lotter AP. Solubility and dissolution properties of generic rifampicin raw materials. Drug Dev Ind Pharm. 2000;26:403–8.PubMedCrossRefGoogle Scholar
  61. 61.
    Agrawal S, Ashokraj Y, Bharatam PV, Pillai O, Panchagnula R. Solid-state characterization of rifampicin samples and its biopharmaceutic relevance. Eur J Pharm Sci. 2004;22:127–44.PubMedCrossRefGoogle Scholar
  62. 62.
    Katzung BG. Basic & clinical pharmacology. 9th ed. New York: Lange Medical Books/McGraw Hill; 2004.Google Scholar
  63. 63.
    CDC. Clinical growth charts. Atlanta: Centers for Disease Control and Prevention, National Center for Health Statistics; 2009. Accessed 31 Aug 2012.
  64. 64.
    Acocella G, Buniva G, Flauto U, Nicolis FB. Absorption and elimination of the antibiotic rifampicin in newborns and children. Proc. 6th Int Congress Chemother. 1969;2:755–60.Google Scholar
  65. 65.
    Koup JR, Williams-Warren J, Viswanathan CT, Weber A, Smith AL. Pharmacokinetics of rifampin in children. II. Oral bioavailability. Ther Drug Monit. 1986;8:17–22.PubMedCrossRefGoogle Scholar
  66. 66.
    Schaaf HS, Parkin DP, Seifart HI, Werely CJ, Hesseling PB, van Helden PD, et al. Isoniazid pharmacokinetics in children treated for respiratory tuberculosis. Arch Dis Child. 2005;90:614–8.PubMedCrossRefGoogle Scholar
  67. 67.
    Ramachandran G, Kumar AK, Swaminathan S. Pharmacokinetics of anti-tuberculosis drugs in children. Indian J Pediatr. 2011;78:435–42.PubMedCrossRefGoogle Scholar
  68. 68.
    Tan TQ, Mason EO, Ou CN, Kaplan SL. Use of intravenous rifampin in neonates with persistent staphylococcal bacteremia. Antimicrob Agents Chemother. 1993;37:2401–6.PubMedCrossRefGoogle Scholar
  69. 69.
    Chik Z, Basu RC, Pendek R, Lee TC, Mohamed Z. A bioequivalence comparison of two formulations of rifampicin (300- vs 150-mg capsules): An open-label, randomized, two-treatment, two-way crossover study in healthy volunteers. Clin Ther. 2010;32:1822–31.PubMedCrossRefGoogle Scholar
  70. 70.
    Cohn HD. Clinical studies with a new rifamycin derivative. J Clin Pharmacol J New Drugs. 1969;9:118–25.PubMedGoogle Scholar
  71. 71.
    Lave T, Luttringer O, Poulin P, Parrott N. Interspecies scaling. In: Krishna R, editor. Applications of pharmacokinetic principles in drug development. New York: Kluwer Academic/Plenum; 2004. p. 133–69.CrossRefGoogle Scholar
  72. 72.
    Webster J, Bollen P, Grimm H, Jennings M. Ethical implications of using the minipig in regulatory toxicology studies. J Pharmacol Toxicol Methods. 2010;62:160–6.PubMedCrossRefGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2013

Authors and Affiliations

  • Wyatt J. Roth
    • 1
  • Candice B. Kissinger
    • 1
  • Robyn R. McCain
    • 2
    • 3
  • Bruce R. Cooper
    • 2
  • Jeremy N. Marchant-Forde
    • 3
    • 4
  • Rachel C. Vreeman
    • 5
  • Sophia Hannou
    • 1
  • Gregory T. Knipp
    • 1
    • 2
    • 3
    Email author
  1. 1.Department of Industrial and Physical Pharmacy, College of PharmacyPurdue UniversityWest LafayetteUSA
  2. 2.Bindley Bioscience CenterPurdue UniversityWest LafayetteUSA
  3. 3.Purdue Translational Pharmacology CTSI Core FacilityPurdue UniversityWest LafayetteUSA
  4. 4.Livestock Behavior Research UnitUSDA-ARSWest LafayetteUSA
  5. 5.Department of PediatricsIndiana University School of MedicineIndianapolisUSA

Personalised recommendations