Pharmaceutical Research

, Volume 34, Issue 6, pp 1320–1329

Structure-Function Analysis of Phenylpiperazine Derivatives as Intestinal Permeation Enhancers

  • Katherine C. Fein
  • Nicholas G. Lamson
  • Kathryn A. Whitehead
Research Paper

Abstract

Purpose

A major obstacle preventing oral administration of macromolecular therapeutics is poor absorption across the intestinal epithelium into the bloodstream. One strategy to improve transport across this barrier is the use of chemical permeation enhancers. Several molecular families with permeation enhancing potential have been identified previously, including piperazines. In particular, 1-phenylpiperazine has been shown to enhance transepithelial transport with minimal cytotoxicity compared to similarly effective molecules. To better understand how the chemistry of 1-phenylpiperazine affects its utility as an intestinal permeation enhancer, this study examined a small library of 13 derivatives of 1-phenylpiperazine.

Methods

The efficacy and cytotoxicity of 13 phenylpiperazine compounds were assessed in a Caco-2 model of the intestinal epithelium. Efficacy was measured using the paracellular diffusion marker calcein as well as by immunostaining and confocal imaging of Caco-2 monolayers.

Results

Of the 13 derivatives, two enhanced the permeability of the fluorescent marker calcein over 100-fold. It was found that hydroxyl or primary amine substitutions on the phenyl ring significantly increased toxicity, while aliphatic substitutions resulted in efficacy and toxicity profiles comparable to 1-phenylpiperazine.

Conclusions

Several potent derivatives, including 1-methyl-4-phenylpiperazine and 1-(4-methylphenyl)piperazine, displayed lower toxicity than 1-phenylpiperazine, suggesting promise in future applications.

Key Words

1-phenylpiperazine Caco-2 oral delivery permeation enhancer piperazine derivatives 

Abbreviations

BSA

Bovine serum albumin

BSM

Basal seeding medium

DMEM

Dulbecco’s Modified Eagle’s Medium

EDM

Enterocyte differentiation medium

EP

Enhancement potential

MTT

Methyl thiazole tetrazolium

OP

Overall potential

TEER

Trans-epithelial electrical resistance

TP

Toxicity potential

Supplementary material

11095_2017_2149_MOESM1_ESM.docx (22 kb)
Table S1(DOCX 21 kb)
11095_2017_2149_MOESM2_ESM.docx (86 kb)
Figure S1(DOCX 86 kb)

References

  1. 1.
    Moroz E, Matoori S, Leroux J-C. Oral delivery of macromolecular drugs: where we are after almost 100 years of attempts. Adv Drug Deliv Rev Elsevier BV. 2016; http://linkinghub.elsevier.com/retrieve/pii/S0169409X16300278
  2. 2.
    Sonaje K, Chen YJ, Chen HL, Wey SP, Juang JH, Nguyen HN, et al. Enteric-coated capsules filled with freeze-dried chitosan/poly(γ-glutamic acid) nanoparticles for oral insulin delivery. Biomaterials Elsevier Ltd. 2010;31(12):3384–94. doi:10.1016/j.biomaterials.2010.01.042.CrossRefGoogle Scholar
  3. 3.
    Fuhrmann G, Grotzky A, Lukić R, Matoori S, Luciani P, Yu H, et al. Sustained gastrointestinal activity of dendronized polymer–enzyme conjugates. Nat Chem. 2013;5(7):582–9. doi:10.1038/nchem.1675.CrossRefPubMedGoogle Scholar
  4. 4.
    Guggi D, Kast CE, Bernkop-Schnürch A. In vivo evaluation of an oral Salmon calcitonin-delivery system based on a Thiolated chitosan carrier matrix. Pharm Res 2003;20(12):1989–1994. http://download.springer.com/static/pdf/545/art%3A10.1023%2FB%3APHAM.0000008047.82334.7d.pdf?originUrl, http://link.springer.com/article/10.1023/B:PHAM.0000008047.82334.7d&token2=exp=1475174816~acl=/static/pdf/545/art%253A10.1023%252FB%253APHAM.0000008047.
  5. 5.
    Aungst BJ. Absorption enhancers: applications and advances. AAPS J. 2012;14(1):10–8.CrossRefPubMedGoogle Scholar
  6. 6.
    Suzuki T. Regulation of intestinal epithelial permeability by tight junctions. Cell Mol Life Sci. 2013;70(4):631–59. doi:10.1007/s00018-012-1070-x.
  7. 7.
    Gupta V, Hwang BH, Lee J, Anselmo AC, Doshi N, Mitragotri S. Mucoadhesive intestinal devices for oral delivery of salmon calcitonin. J Control Release Elsevier BV. 2013;172(3):753–62. http://www.ncbi.nlm.nih.gov/pubmed/24035976 CrossRefGoogle Scholar
  8. 8.
    Whitehead K, Shen Z, Mitragotri S. Oral delivery of macromolecules using intestinal patches: applications for insulin delivery. J Control Release. 2004;98(1):37–45.CrossRefPubMedGoogle Scholar
  9. 9.
    Lowman a M, Morishita M, Kajita M, Nagai T, Peppas N a. Oral delivery of insulin using pH-responsive complexation gels. J Pharm Sci. 1999;88(9):933–7.CrossRefPubMedGoogle Scholar
  10. 10.
    Pridgen EM, Alexis F, Kuo TT, Levy-Nissenbaum E, Karnik R, Blumberg RS, et al. Transepithelial transport of Fc-targeted nanoparticles by the neonatal fc receptor for oral delivery. Sci Transl Med. 2013;5(213):213ra167. http://stm.sciencemag.org/content/5/213/213ra167.short CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Salama NN, Eddington ND, Fasano A. Tight junction modulation and its relationship to drug delivery. Adv Drug Deliv Rev. 2006;58(1):15–28.CrossRefPubMedGoogle Scholar
  12. 12.
    Gupta V, Hwang BH, Doshi N, Mitragotri S. A permeation enhancer for increasing transport of therapeutic macromolecules across the intestine. J Control Release. Elsevier B.V. 2013;172:541–9. doi:10.1016/j.jconrel.2013.05.002.CrossRefGoogle Scholar
  13. 13.
    Lindmark T, Kimura YAP, Lindmark T, Kimura Y, Artursson P. Absorption enhancement through intracellular regulation of tight junction permeability by medium chain fatty acids in Caco-2 cells. J Pharmacol Exp Ther. 1998;284(1):362–9. http://www.ncbi.nlm.nih.gov/pubmed/9435199 Google Scholar
  14. 14.
    Sonaje K, Chuang EY, Lin KJ, Yen TC, Su FY, Tseng MT, et al. Opening of epithelial tight junctions and enhancement of paracellular permeation by chitosan: microscopic, ultrastructural, and computed-tomographic observations. Mol Pharm. 2012;9(5):1271–9.PubMedGoogle Scholar
  15. 15.
    Fasano a, Baudry B, Pumplin DW, Wasserman SS, Tall BD, Ketley JM, et al. Vibrio cholerae produces a second enterotoxin, which affects intestinal tight junctions. Proc Natl Acad Sci U S A. 1991;88(12):5242–6.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Whitehead K, Karr N, Mitragotri S. Safe and effective permeation enhancers for oral drug delivery. Pharm Res. 2008;25:1782–8.CrossRefPubMedGoogle Scholar
  17. 17.
    Lamson NG, Cusimano G, Suri K, Zhang A, Whitehead KA. The pH of Piperazine derivative solutions predicts their utility as Transepithelial permeation enhancers. Mol Pharm. 2016;13(2):578–85. doi:10.1021/acs.molpharmaceut.5b00803.CrossRefPubMedGoogle Scholar
  18. 18.
    Bzik VA, Brayden DJ. An assessment of the permeation enhancer, 1-phenyl-piperazine (PPZ), on Paracellular flux across rat intestinal mucosae in Ussing chambers. Pharm Res Pharmaceutical Research. 2016;33(10):2506–16. doi:10.1007/s11095-016-1975-4.PubMedGoogle Scholar
  19. 19.
    Sambuy Y, De Angelis I, Ranaldi G, Scarino ML, Stammati A, Zucco F. The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics. Cell Biol Toxicol. 2005;21(1):1–26.CrossRefPubMedGoogle Scholar
  20. 20.
    Artursson PER. Epithelial transport of drugs in cell culture. I : a model for studying the passive diffusion of drugs over intestinal. J Pharm Sci. 1989;79(6):476–82.CrossRefGoogle Scholar
  21. 21.
    Srinivasan B, Kolli AR, Esch MB, Abaci HE, Shuler ML, Hickman JJ. TEER measurement techniques for in vitro barrier model systems. J Lab Autom. 2015;20(2):107–26. http://www.ncbi.nlm.nih.gov/pubmed/25586998 CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Shaquiquzzaman M, Verma G, Marella A, Akhter M, Akhtar W, Khan MF, et al. European Journal of medicinal chemistry Piperazine scaffold : a remarkable tool in generation of diverse pharmacological agents relative light units. Eur J Med Chem Elsevier Masson SAS. 2015;102:487–529. doi:10.1016/j.ejmech.2015.07.026.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Katherine C. Fein
    • 1
  • Nicholas G. Lamson
    • 1
  • Kathryn A. Whitehead
    • 1
    • 2
  1. 1.Department of Chemical EngineeringCarnegie Mellon UniversityPittsburghUSA
  2. 2.Department of Biomedical EngineeringPittsburghUSA

Personalised recommendations