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A Mouse Model to Evaluate the Impact of Species, Sex, and Lipid Load on Lymphatic Drug Transport

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ABSTRACT

Purpose

To establish a lymph-cannulated mouse model, and use the model to investigate the impact of lipid dose on exogenous and endogenous lipid recruitment, and drug transport, into the lymph of males versus females. Finally, lymphatic transport and drug absorption in the mouse were compared to other pre-clinical models (rats/dogs).

Methods

Animals were orally or intraduodenally administered 1.6 mg/kg halofantrine in low or high 14C-lipid doses. For bioavailability calculation, animals were intravenuosly administered halofantrine. Lymph or blood samples were taken and halofantrine, triglyceride, phospholipid and 14C-lipid concentrations measured.

Results

Lymphatic lipid transport increased linearly with lipid dose, was similar across species and in male/female animals. In contrast, lymphatic transport of halofantrine differed markedly across species (dogs>rats>mice) and plateaued at higher lipid doses. Lower bioavailability appeared responsible for some species differences in halofantrine lymphatic transport; however other systematic differences were involved.

Conclusions

A contemporary lymph-cannulated mouse model was established which will enable investigation of lymphatic transport in transgenic and disease models. The current study found halofantrine absorption and lymphatic transport are reduced in small animals. Future analyses will investigate mechanisms involved, and if similar trends occur for other drugs, to establish the most relevant model(s) to predict lymphatic transport in humans.

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Abbreviations

AUC:

Area under the plasma concentration-time curve

FA:

Fatty acid

Hf:

Halofantrine

HPLC:

High Performance Liquid Chromatography

LC-MS:

HPLC-mass spectrometry

PL:

Phospholipid

SEDDS:

Self-Emulsifying Drug Delivery System

TG:

Triglyceride

REFERENCES

  1. Williams HD, Trevaskis NL, Charman SA, Shanker RM, Charman WN, Pouton CW, et al. Strategies to address low drug solubility in discovery and development. Pharmacol Rev. 2013;65:315–499.

    Google Scholar 

  2. Charman WN, Stella VJ. Estimating the maximum potential for intestinal lymphatic transport of lipophilic drug molecules. Int J Pharm. 1986;34:175–8.

    Article  CAS  Google Scholar 

  3. Gershkovich P, Hoffman A. Uptake of lipophilic drugs by plasma derived isolated chylomicrons: linear correlation with intestinal lymphatic bioavailability. Eur J Pharm Sci. 2005;26:394–404.

    Article  PubMed  CAS  Google Scholar 

  4. Trevaskis NL, McEvoy CL, McIntosh MP, Edwards GA, Shanker RM, Charman WN, et al. The role of the intestinal lymphatics in the absorption of two highly lipophilic cholesterol ester transfer protein inhibitors (CP524,515 and CP532,623). Pharm Res. 2010;27:878–93.

    Article  PubMed  CAS  Google Scholar 

  5. Trevaskis NL, Shanker RM, Charman WN, Porter CJ. The mechanism of lymphatic access of two cholesteryl ester transfer protein inhibitors (CP524,515 and CP532,623) and evaluation of their impact on lymph lipoprotein profiles. Pharm Res. 2010;27:1949–64.

    Article  PubMed  CAS  Google Scholar 

  6. Porter CJ, Trevaskis NL, Charman WN. Lipids and lipid-based formulations: optimizing the oral delivery of lipophilic drugs. Nat Rev Drug Discov. 2007;6:231–48.

    Article  PubMed  CAS  Google Scholar 

  7. Trevaskis NL, Charman WN, Porter CJ. Lipid-based delivery systems and intestinal lymphatic drug transport: a mechanistic update. Adv Drug Deliv Rev. 2008;60:702–16.

    Article  PubMed  CAS  Google Scholar 

  8. Trevaskis NL, Porter CJ, Charman WN. An examination of the interplay between enterocyte-based metabolism and lymphatic drug transport in the rat. Drug Metab Dispos. 2006;34:729–33.

    Article  PubMed  CAS  Google Scholar 

  9. Khoo SM, Edwards GA, Porter CJ, Charman WN. A conscious dog model for assessing the absorption, enterocyte-based metabolism, and intestinal lymphatic transport of halofantrine. J Pharm Sci. 2001;90:1599–607.

    Article  PubMed  CAS  Google Scholar 

  10. Trevaskis NL, Shackleford DM, Charman WN, Edwards GA, Gardin A, Appel-Dingemanse S, et al. Intestinal lymphatic transport enhances the post-prandial oral bioavailability of a novel cannabinoid receptor agonist via avoidance of first-pass metabolism. Pharm Res. 2009;26:1486–95.

    Article  PubMed  CAS  Google Scholar 

  11. Shackleford DM, Faassen WA, Houwing N, Lass H, Edwards GA, Porter CJ, et al. Contribution of lymphatically transported testosterone undecanoate to the systemic exposure of testosterone after oral administration of two andriol formulations in conscious lymph duct-cannulated dogs. J Pharmacol Exp Ther. 2003;306:925–33.

    Article  PubMed  CAS  Google Scholar 

  12. White KL, Nguyen G, Charman WN, Edwards GA, Faassen WA, Porter CJ. Lymphatic transport of Methylnortestosterone undecanoate (MU) and the bioavailability of methylnortestosterone are highly sensitive to the mass of coadministered lipid after oral administration of MU. J Pharmacol Exp Ther. 2009;331:700–9.

    Article  PubMed  CAS  Google Scholar 

  13. Caliph SM, Trevaskis NL, Charman WN, Porter CJ. Intravenous dosing conditions may affect systemic clearance for highly lipophilic drugs: implications for lymphatic transport and absolute bioavailability studies. J Pharm Sci. 2012;101:3540–6.

    Article  PubMed  CAS  Google Scholar 

  14. Hauss DJ, Mehta SC, Radebaugh GW. Targeted lymphatic transport and modified systemic distribution of CI-976, a lipophilic lipid-regulator drug, via a formulation approach. Int J Pharm. 1994;108:85–93.

    Article  CAS  Google Scholar 

  15. Trevaskis NL, Charman WN, Porter CJ. Targeted drug delivery to lymphocytes: a route to site-specific immunomodulation? Mol Pharm. 2010;7:2297–309.

    Article  PubMed  CAS  Google Scholar 

  16. Dane KY, Nembrini C, Tomei AA, Eby JK, O’Neil CP, Velluto D, et al. Nano-sized drug-loaded micelles deliver payload to lymph node immune cells and prolong allograft survival. J Control Release. 2011;156:154–60.

    Article  PubMed  CAS  Google Scholar 

  17. Kaminskas LM, Porter CJH. Targeting the lymphatics using dendritic polymers (dendrimers). Adv Drug Deliv Rev. 2011;63:890–900.

    Article  PubMed  CAS  Google Scholar 

  18. Caliph SM, Faassen WA, Vogel GM, Porter CJ. Oral bioavailability assessment and intestinal lymphatic transport of Org 45697 and Org 46035, two highly lipophilic novel immunomodulator analogues. Curr Drug Deliv. 2009;6:359–66.

    Article  PubMed  CAS  Google Scholar 

  19. Holm R, Mullertz A, Christensen E, Hoy CE, Kristensen HG. Comparison of total oral bioavailability and the lymphatic transport of halofantrine from three different unsaturated triglycerides in lymph-cannulated conscious rats. Eur J Pharm Sci. 2001;14:331–7.

    Article  PubMed  CAS  Google Scholar 

  20. Porter CJ, Charman SA, Humberstone AJ, Charman WN. Lymphatic transport of halofantrine in the conscious rat when administered as either the free base or the hydrochloride salt: effect of lipid class and lipid vehicle dispersion. J Pharm Sci. 1996;85:357–61.

    Article  PubMed  CAS  Google Scholar 

  21. Trevaskis NL, Porter CJ, Charman WN. Bile increases intestinal lymphatic drug transport in the fasted rat. Pharm Res. 2005;22:1863–70.

    Article  PubMed  CAS  Google Scholar 

  22. Dahan A, Mendelman A, Amsili S, Ezov N, Hoffman A. The effect of general anesthesia on the intestinal lymphatic transport of lipophilic drugs: comparison between anesthetized and freely moving conscious rat models. Eur J Pharm Sci. 2007;32:367–74.

    Article  PubMed  CAS  Google Scholar 

  23. Lespine A, Chanoit G, Bousquet-Melou A, Lallemand E, Bassissi FM, Alvinerie M, et al. Contribution of lymphatic transport to the systemic exposure of orally administered moxidectin in conscious lymph duct-cannulated dogs. Eur J Pharm Sci. 2006;27:37–43.

    Article  PubMed  CAS  Google Scholar 

  24. Lo CM, Nordskog BK, Nauli AM, Zheng S, Vonlehmden SB, Yang Q, et al. Why does the gut choose apolipoprotein B48 but not B100 for chylomicron formation? Am J Physiol Gastrointest Liver Physiol. 2008;294:G344–52.

    Article  PubMed  CAS  Google Scholar 

  25. Nauli AM, Nassir F, Zheng S, Yang Q, Lo CM, Vonlehmden SB, et al. CD36 is important for chylomicron formation and secretion and may mediate cholesterol uptake in the proximal intestine. Gastroenterology. 2006;131:1197–207.

    Article  PubMed  CAS  Google Scholar 

  26. Caliph SM, Charman WN, Porter CJ. Effect of short-, medium-, and long-chain fatty acid-based vehicles on the absolute oral bioavailability and intestinal lymphatic transport of halofantrine and assessment of mass balance in lymph-cannulated and non-cannulated rats. J Pharm Sci. 2000;89:1073–84.

    Article  PubMed  CAS  Google Scholar 

  27. Porter CJ, Charman SA, Charman WN. Lymphatic transport of halofantrine in the triple-cannulated anesthetized rat model: effect of lipid vehicle dispersion. J Pharm Sci. 1996;85:351–6.

    Article  PubMed  CAS  Google Scholar 

  28. Trevaskis NL, Porter CJ, Charman WN. The lymph lipid precursor pool is a key determinant of intestinal lymphatic drug transport. J Pharmacol Exp Ther. 2006;316:881–91.

    Article  PubMed  CAS  Google Scholar 

  29. Khoo SM, Shackleford DM, Porter CJ, Edwards GA, Charman WN. Intestinal lymphatic transport of halofantrine occurs after oral administration of a unit-dose lipid-based formulation to fasted dogs. Pharm Res. 2003;20:1460–5.

    Article  PubMed  CAS  Google Scholar 

  30. Khoo SM, Humberstone AJ, Porter CJ, Edwards GA, Charman WN. Formulation design and bioavailability assessment of lipidic self-emulsifying formulations of halofantrine. Int J Pharm. 1998;167:155–64.

    Article  CAS  Google Scholar 

  31. Johnson BM, Chen W, Borchardt RT, Charman WN, Porter CJ. A kinetic evaluation of the absorption, efflux, and metabolism of verapamil in the autoperfused rat jejunum. J Pharmacol Exp Ther. 2003;305:151–8.

    Article  PubMed  CAS  Google Scholar 

  32. Edwards GA, Porter CJ, Caliph SM, Khoo SM, Charman WN. Animal models for the study of intestinal lymphatic drug transport. Adv Drug Deliv Rev. 2001;50:45–60.

    Article  PubMed  CAS  Google Scholar 

  33. Humberstone AJ, Currie GJ, Porter CJ, Scanlon MJ, Charman WN. A simplified liquid chromatography assay for the quantitation of halofantrine and desbutylhalofantrine in plasma and identification of a degradation product of desbutylhalofantrine formed under alkaline conditions. J Pharm Biomed Anal. 1995;13:265–72.

    Article  PubMed  CAS  Google Scholar 

  34. Bauer R, Guzy S, Ng C. A survey of population analysis methods and software for complex pharmacokinetic and pharmacodynamic models with examples. AAPS J. 2007;9:E60–83.

    Article  PubMed  Google Scholar 

  35. Bulitta JB, Bingolbali A, Shin BS, Landersdorfer CB. Development of a new pre- and post-processing tool (SADAPT-TRAN) for nonlinear mixed-effects modeling in S-ADAPT. AAPS J. 2011;13:201–11.

    Article  PubMed  Google Scholar 

  36. Prokocimer P, Bien P, Surber J, Mehra P, DeAnda C, Bulitta JB, et al. Phase 2, randomized, double-blind, dose-ranging study evaluating the safety, tolerability, population pharmacokinetics, and efficacy of oral torezolid phosphate in patients with complicated skin and skin structure infections. Antimicrob Agents Chemother. 2011;55:583–92.

    Article  PubMed  CAS  Google Scholar 

  37. Bulitta JB, Kinzig M, Landersdorfer CB, Holzgrabe U, Stephan U, Sorgel F. Comparable population pharmacokinetics and pharmacodynamic breakpoints of cefpirome in cystic fibrosis patients and healthy volunteers. Antimicrob Agents Chemother. 2011;55:2927–36.

    Article  PubMed  CAS  Google Scholar 

  38. Bailer AJ. Testing for the equality of area under the curves when using destructive measurement techniques. J Pharmacokinet Biopharm. 1988;16:303–9.

    Article  PubMed  CAS  Google Scholar 

  39. Kohan AB, Yoder SM, Tso P. Using the lymphatics to study nutrient absorption and the secretion of gastrointestinal hormones. Physiol Behav. 2011;105:82–8.

    Article  PubMed  CAS  Google Scholar 

  40. Mansbach CM, Arnold A. Steady-state kinetic analysis of triacylglycerol delivery into mesenteric lymph. Am J Physiol. 1986;251:G263–9.

    PubMed  CAS  Google Scholar 

  41. Mansbach II CM, Dowell RF, Pritchett D. Portal transport of absorbed lipids in rats. Am J Physiol. 1991;261:G530–8.

    PubMed  CAS  Google Scholar 

  42. McDonald GB, Weidman M. Partitioning of polar fatty acids into lymph and portal vein after intestinal absorption in the rat. Q J Exp Physiol. 1987;72:153–9.

    PubMed  CAS  Google Scholar 

  43. Drover VA, Ajmal M, Nassir F, Davidson NO, Nauli AM, Sahoo D, et al. CD36 deficiency impairs intestinal lipid secretion and clearance of chylomicrons from the blood. J Clin Invest. 2005;115:1290–7.

    PubMed  CAS  Google Scholar 

  44. Nauli AM. Intestinal lipid uptake and secretion of VLDL and chylomicron, Department of Pathology and Laboratory Medicine, Ph D thesis, University of Cincinnati, Cincinnati; 2005. p. 170.

  45. Vahouny GV, Blendermann EM, Gallo LL, Treadwell CR. Differential transport of cholesterol and oleic acid in lymph lipoproteins: sex differences in puromycin sensitivity. J Lipid Res. 1980;21:415–24.

    PubMed  CAS  Google Scholar 

  46. Krause BR, Sloop CH, Castle CK, Roheim PS. Mesenteric lymph apolipoproteins in control and ethinyl estradiol-treated rats: a model for studying apolipoproteins of intestinal origin. J Lipid Res. 1981;22:610–9.

    PubMed  CAS  Google Scholar 

  47. Yang L, Koo SI, Jeon IJ. The lymphatic absorption of fatty acids and output of phospholipids are lowered by estrogen replacement in ovariectomized rats. Nutr Biochem. 1996;7:214–21.

    Article  CAS  Google Scholar 

  48. Chaikoff IL, Bloom B, Stevens BP, Reinhardt WO, Dauben WG. Pentadecanoic acid-5-C14; its absorption and lymphatic transport. J Biol Chem. 1951;190:431–5.

    PubMed  CAS  Google Scholar 

  49. Bloom B, Chaikoff IL, Reinhardt. Intestinal lymph as pathway for transport of absorbed fatty acids of different chain lengths. Am J Physiol. 1951;166:451–5.

    PubMed  CAS  Google Scholar 

  50. Kiyasu JY, Bloom B, Chaikoff IL. The portal transport of absorbed fatty acids. J Biol Chem. 1952;199:415–9.

    PubMed  CAS  Google Scholar 

  51. Trevaskis NL, Charman WN, Porter CJ. Acute hypertriglyceridemia promotes intestinal lymphatic lipid and drug transport: a positive feedback mechanism in lipid and drug absorption. Mol Pharm. 2011;8:1132–9.

    Article  PubMed  CAS  Google Scholar 

  52. Shiau YF, Popper DA, Reed M, Umstetter C, Capuzzi D, Levine GM. Intestinal triglycerides are derived from both endogenous and exogenous sources. Am J Physiol. 1985;248:G164–9.

    PubMed  CAS  Google Scholar 

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ACKNOWLEDGMENTS AND DISCLOSURES

The authors would like to thank Luojuan Hu for her technical assistance in the mouse bioavailability experiments and Dr Juergen Bulitta for assistance in calculating bioavailability in the mice.

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Correspondence to Natalie L. Trevaskis or Christopher J. H. Porter.

Electronic supplementary material

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Supplementary figure 1

Panel A: Cumulative% of the dose of halofantrine (Hf), and Panel B: Cumulative triglyceride (TG, mg) mass, transported in the mesenteric lymph following administration of 1.6 mg/kg halofantrine and 18.1 mg/kg long chain lipid, as an oleic acid emulsion administered into the intestine of anaesthetised rats (●) or as a pre-dispersed self-emulsifying drug delivery system via oral administration to conscious rats (○). Data represent mean ± SEM for n = 3–4 animals. (DOC 91 kb)

Supplementary figure 2

Panel A. Mass of total, endogenous and exogenous fatty acid transported into the lymph of male (black bars) or female (white bars) rats, and Panel B. Cumulative% of the dose of halofantrine transported in the lymph of male (●) or female (○) rats, over 8 h following intraduodenal administration of 1.6 mg/kg halofantrine in an emulsion containing 18.1 mg/kg long chain lipid. Data represent mean ± SEM for n = 3–4 animals. *Parameter significantly greater in male when compared to female animals (α < 0.05). (DOC 93 kb)

Supplementary figure 3

Plasma halofantrine (Hf) concentration vs time profiles in mice (Panel A) and rats (Panel B) following administration of 1.6 mg/kg halofantrine and 18.1 mg/kg long chain lipid as an oleic acid emulsion administered into the intestine of anaesthetised animals (●) or as a pre-dispersed self-emulsifying drug delivery system via oral administration to conscious animals (○). Data represent mean ± SEM for n = 3–6 animals. (DOC 101 kb)

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Trevaskis, N.L., Caliph, S.M., Nguyen, G. et al. A Mouse Model to Evaluate the Impact of Species, Sex, and Lipid Load on Lymphatic Drug Transport. Pharm Res 30, 3254–3270 (2013). https://doi.org/10.1007/s11095-013-1000-0

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