Abstract
Purpose. The transport of peptides or proteins across the alveolar cell monolayer was studied in vitro in order to elucidate their transport pathway.
Methods. The permeability of 14 peptides or proteins and 6 dextrans with MW 1,000~150,000 was measured in cultured human lung adenocarcinoma A549 cell monolayers at 37°C or 4°C. The stability of the tested peptides and proteins was also evaluated.
Results. The permeability coefficients of these macromolecules across the A549 cell monolayer at 37°C ranged from 10 −5 to 10 −7 (cm/sec), and exhibited a good inverse correlation with molecular weight. All macromolecules were stable throughout the transport experiment, and degradation by proteases was minimal. Permeability at 4°C did not differ from that at 37°C. Clear selectivity for direction of transport was not observed.
Conclusions. These results suggested that the tested peptides and proteins appeared to penetrate the A549 cell monolayer via a paracellular route by passive diffusion.
Similar content being viewed by others
REFERENCES
J.S. Patton and R.M. Platz. (D) Routes of delivery: Case studies, (2) Pulmonary delivery of peptides and proteins for systemic action. Adv. Drug Deliv. Rev. 8: 179–196 (1992).
D.T. O'Hagan and L. Illum. Absorption of peptides and proteins from the respiratory tract and the potential for development of locally administered vaccine. Critical Reviews in Therapeutic Drug Carrier Systems 7, Issue 1, 35–97 (1990).
T. Fujita, A. Yamamoto and S. Muranishi. Comparison of permeability of macromolecular drugs across cultured intestinal and alveolar epithelial cell lines. International Symposium. Delivery of Protein Drugs — The Next 10 Years, September 4–9, 1993 Koyto, Japan.
H. Yamahara, C.M. Lehr, V.H.L. Lee and K.J. Kim. Fate of insulin during transit across rat alveolar epithelial cell monolayers. Eur. J. Pharm. Biopharm. 40: 294–298 (1994).
M. Lieber, B. Smith, A. Szakal, W. Nelson-Rees and G. Todaro. A continuous tumor-cell line from a human lung carcinoma with properties of type II alveolar epithelial cells. Int. J. Cancer 17: 62–70 (1976).
D.L. Shapiro, L.L. Nardone, S.A. Rooney, E.K. Motoyama and J.L. Munoz. Phospholipid biosynthesis and secretion by a cell line (A549) which resembles type II alveolar epithelial cells. Biochim. Biophys. Acta 530: 197–207 (1978).
S.J. Enna and L.S. Schanker. Absorption of saccharides and urea from the rat lung. Am. J. Physiol. 222: 409–414 (1972).
L.G. Johnson, P.W. Cheng and R.C. Boucher. Albumin absorption by canine bronchial epithelium. Am. J. Appl. Physiol. 66: 2772–2777 (1989).
J. Theodore, E.D. Robin, R. Gaudio and J. Acevedo. Transalveolar transport of large polar solutes (sucrose, inulin, and dextran). Am. J. Physiol. 229: 989–996 (1975).
A.B. Gorin and P.A. Stewart. Differential permeability of endothelial and epithelial barriers to albumin flux. J. Appl. Physiol. 47: 1315–1324 (1979).
P.R. Byron and E.M. Phillips. Chapt. 5 Absorption, clearance, and dissolution in the lung, p. 112. Respiratory Drug Delivery, CRC Press, Florida (1990).
J.M. Mullin and M.T. McGinn. Effects of diacylglycerols on LLC-PK1 renal epithelia: Similarity to phorbol ester tumor promoters. J. Cell. Physiol. 134: 357–366 (1988).
D.T. Thwaites, B.H. Hirst and N.L. Simmons. Passive transepithelial absorption of thyrotropin-releasing hormone (TRH) via a paracellular route in cultured intestinal and renal epithelial cell lines. Pharm. Res. 10: 674–681 (1993).
E.E. Schneeberger. Heterogeneity of tight junction morphology in extrapulmonary and intrapulmonary airways of the rat. Anat. Rec. 198: 193–208 (1980).
R.M. Effros and G.R. Mason. Measurements of pulmonary epithelial permeability in vivo. Am. Rev. Resp. Dis. 127, S59–S61 (1983).
L.S. Schanker and J.A. Hemberger. Relation between molecular weight and pulmonary absorption rate of lipid-insoluble compounds in neonatal and adult rats. Biochem. Pharmacol. 32: 2599–2601 (1983).
A.N. Fisher, K. Brown, S.S. Davis, G.D. Parr and D.A. Smith. The effect of molecular size on the nasal absorption of water-soluble compounds in the albino rat. J. Pharm. Pharmacol. 39: 357–362 (1987).
J.S. Patton. T. Trinchero and R.M. Platz. Bio-availability of pulmonary delivered peptides and proteins: α-interferon, calcitonins and parathyroid hormones. J. Contr. Release 28: 79–85 (1994).
R.W. Niven, F. Rypacek and P.R. Byron. Solute absorption from the airways of the isolated rat lung. III. Absorption of several peptidase-resistant, synthetic polypeptidases: Poly-(2-hydroxyethyl)-aspartamides. Pharm. Res. 7: 990–994 (1990).
S. Kobayashi, S. Kondo and K. Juni. Study on pulmonary delivery of salmon calcitonin in rats: Effects of protease inhibitors and absorption enhancers. Pharm. Res. 11, 1239–1243 (1994).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kobayashi, S., Kondo, S. & Juni, K. Permeability of Peptides and Proteins in Human Cultured Alveolar A549 Cell Monolayer. Pharm Res 12, 1115–1119 (1995). https://doi.org/10.1023/A:1016295406473
Issue Date:
DOI: https://doi.org/10.1023/A:1016295406473