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Paracellular permeation-enhancing effect of AT1002 C-terminal amidation in nasal delivery.

Song KH, Kim SB, Shim CK, Chung SJ, Kim DD, Rhee SK, Choi GJ, Kim CH, Kim K - Drug Des Devel Ther (2015)

Bottom Line: Pep1 was nontoxic to cell monolayers and showed a relatively low decrease in peak area compared to AT1002.In addition, administration of mannitol with Pep1 resulted in significant increases in the area under the plasma concentration-time curve and peak plasma concentration at 3.63-fold and 2.68-fold, respectively, compared to mannitol alone.Thus, Pep1 increased the stability or possibly reduced the instability of AT1002, which resulted in an increased permeation-enhancing effect of AT1002.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Engineering, Soonchunhyang University, Asan, Republic of Korea.

ABSTRACT

Background: The identification of permeation enhancers has gained interest in the development of drug delivery systems. A six-mer peptide, H-FCIGRL-OH (AT1002), is a tight junction modulator with promising permeation-enhancing activity. AT1002 enhances the transport of molecular weight markers or agents with low bioavailability with no cytotoxicity. However, AT1002 is not stable in neutral pH or after incubation under physiological conditions, which is necessary to fully uncover its permeation-enhancing effect. Thus, we increased the stability or mitigated the instability of AT1002 by modifying its terminal amino acids and evaluated its subsequent biological activity.

Methods: C-terminal-amidated (FCIGRL-NH2, Pep1) and N-terminal-acetylated (Ac-FCIGRL, Pep2) peptides were analyzed by liquid chromatography-mass spectrometry. We further assessed cytotoxicity on cell monolayers, as well as the permeation-enhancing activity following nasal administration of the paracellular marker mannitol.

Results: Pep1 was nontoxic to cell monolayers and showed a relatively low decrease in peak area compared to AT1002. In addition, administration of mannitol with Pep1 resulted in significant increases in the area under the plasma concentration-time curve and peak plasma concentration at 3.63-fold and 2.68-fold, respectively, compared to mannitol alone. In contrast, no increase in mannitol concentration was shown with mannitol/AT1002 or mannitol/Pep2 compared to the control. Thus, Pep1 increased the stability or possibly reduced the instability of AT1002, which resulted in an increased permeation-enhancing effect of AT1002.

Conclusion: These results suggest the potential usefulness of C-terminal-amidated AT1002 in enhancing nasal drug delivery, which may lead to the development of a practical drug delivery technology for drugs with low bioavailability.

No MeSH data available.


Related in: MedlinePlus

Average plasma concentration of [3H]-mannitol versus time in femoral vein-cannulated SD rats following the intranasal administration of each peptide formulation.Notes: Mannitol/FCIGRL-NH2 (Pep1, •), mannitol/Ac-FCIGRL (Pep2, ○), mannitol/FCIGRL (AT1002, ▼), and mannitol alone (Δ). Each data point represents the mean ± SEM of 3–4 rats. 20 μCi/kg of [3H]-mannitol, 2.5 mg/kg of peptide in 5% dextrose solution; **P<0.01.Abbreviations: SD, Sprague Dawley; SEM, standard error of the mean.
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f6-dddt-9-1815: Average plasma concentration of [3H]-mannitol versus time in femoral vein-cannulated SD rats following the intranasal administration of each peptide formulation.Notes: Mannitol/FCIGRL-NH2 (Pep1, •), mannitol/Ac-FCIGRL (Pep2, ○), mannitol/FCIGRL (AT1002, ▼), and mannitol alone (Δ). Each data point represents the mean ± SEM of 3–4 rats. 20 μCi/kg of [3H]-mannitol, 2.5 mg/kg of peptide in 5% dextrose solution; **P<0.01.Abbreviations: SD, Sprague Dawley; SEM, standard error of the mean.

Mentions: The pharmacokinetic profiles were characterized for each formulation, and the parameters were calculated using noncompartmental analysis. The mean (± SEM) plasma concentration versus time profiles following the intranasal administration of mannitol formulations are shown in Figure 6. Significantly higher permeation of mannitol was found in the formulations of mannitol/Pep1 compared to the control formulation of mannitol alone after 40 minutes, indicating a significant enhancement in the intranasal absorption of mannitol with Pep1 (P<0.01). The administration of mannitol with Pep1 produced a significant increase (3.63-fold) in AUC0–360 minutes (22.08±3.54 min ng/mL) and an increase (2.68-fold) in Cmax (82.06±12.10 pg/mL) over the control formulation of mannitol alone (AUC0–360 minutes =6.08±1.33 min ng/mL, Cmax =30.58±6.02 pg/mL; P<0.01; Table 1). However, there were no significant differences in mannitol concentration at each time point between the formulations of mannitol/AT1002, mannitol/Pep2, and mannitol alone. The enhancement of mannitol by Pep1 was in accordance with no statistically significant difference over all of the incubation times in the peak areas of Pep1 compared to AT1002 or Pep2 (Figure 4), suggesting that C-terminal amidation of AT1002 increased the stability or at least reduces the instability of AT1002, which exposed the inherent permeation-enhancing effect of AT1002.


Paracellular permeation-enhancing effect of AT1002 C-terminal amidation in nasal delivery.

Song KH, Kim SB, Shim CK, Chung SJ, Kim DD, Rhee SK, Choi GJ, Kim CH, Kim K - Drug Des Devel Ther (2015)

Average plasma concentration of [3H]-mannitol versus time in femoral vein-cannulated SD rats following the intranasal administration of each peptide formulation.Notes: Mannitol/FCIGRL-NH2 (Pep1, •), mannitol/Ac-FCIGRL (Pep2, ○), mannitol/FCIGRL (AT1002, ▼), and mannitol alone (Δ). Each data point represents the mean ± SEM of 3–4 rats. 20 μCi/kg of [3H]-mannitol, 2.5 mg/kg of peptide in 5% dextrose solution; **P<0.01.Abbreviations: SD, Sprague Dawley; SEM, standard error of the mean.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4383222&req=5

f6-dddt-9-1815: Average plasma concentration of [3H]-mannitol versus time in femoral vein-cannulated SD rats following the intranasal administration of each peptide formulation.Notes: Mannitol/FCIGRL-NH2 (Pep1, •), mannitol/Ac-FCIGRL (Pep2, ○), mannitol/FCIGRL (AT1002, ▼), and mannitol alone (Δ). Each data point represents the mean ± SEM of 3–4 rats. 20 μCi/kg of [3H]-mannitol, 2.5 mg/kg of peptide in 5% dextrose solution; **P<0.01.Abbreviations: SD, Sprague Dawley; SEM, standard error of the mean.
Mentions: The pharmacokinetic profiles were characterized for each formulation, and the parameters were calculated using noncompartmental analysis. The mean (± SEM) plasma concentration versus time profiles following the intranasal administration of mannitol formulations are shown in Figure 6. Significantly higher permeation of mannitol was found in the formulations of mannitol/Pep1 compared to the control formulation of mannitol alone after 40 minutes, indicating a significant enhancement in the intranasal absorption of mannitol with Pep1 (P<0.01). The administration of mannitol with Pep1 produced a significant increase (3.63-fold) in AUC0–360 minutes (22.08±3.54 min ng/mL) and an increase (2.68-fold) in Cmax (82.06±12.10 pg/mL) over the control formulation of mannitol alone (AUC0–360 minutes =6.08±1.33 min ng/mL, Cmax =30.58±6.02 pg/mL; P<0.01; Table 1). However, there were no significant differences in mannitol concentration at each time point between the formulations of mannitol/AT1002, mannitol/Pep2, and mannitol alone. The enhancement of mannitol by Pep1 was in accordance with no statistically significant difference over all of the incubation times in the peak areas of Pep1 compared to AT1002 or Pep2 (Figure 4), suggesting that C-terminal amidation of AT1002 increased the stability or at least reduces the instability of AT1002, which exposed the inherent permeation-enhancing effect of AT1002.

Bottom Line: Pep1 was nontoxic to cell monolayers and showed a relatively low decrease in peak area compared to AT1002.In addition, administration of mannitol with Pep1 resulted in significant increases in the area under the plasma concentration-time curve and peak plasma concentration at 3.63-fold and 2.68-fold, respectively, compared to mannitol alone.Thus, Pep1 increased the stability or possibly reduced the instability of AT1002, which resulted in an increased permeation-enhancing effect of AT1002.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Engineering, Soonchunhyang University, Asan, Republic of Korea.

ABSTRACT

Background: The identification of permeation enhancers has gained interest in the development of drug delivery systems. A six-mer peptide, H-FCIGRL-OH (AT1002), is a tight junction modulator with promising permeation-enhancing activity. AT1002 enhances the transport of molecular weight markers or agents with low bioavailability with no cytotoxicity. However, AT1002 is not stable in neutral pH or after incubation under physiological conditions, which is necessary to fully uncover its permeation-enhancing effect. Thus, we increased the stability or mitigated the instability of AT1002 by modifying its terminal amino acids and evaluated its subsequent biological activity.

Methods: C-terminal-amidated (FCIGRL-NH2, Pep1) and N-terminal-acetylated (Ac-FCIGRL, Pep2) peptides were analyzed by liquid chromatography-mass spectrometry. We further assessed cytotoxicity on cell monolayers, as well as the permeation-enhancing activity following nasal administration of the paracellular marker mannitol.

Results: Pep1 was nontoxic to cell monolayers and showed a relatively low decrease in peak area compared to AT1002. In addition, administration of mannitol with Pep1 resulted in significant increases in the area under the plasma concentration-time curve and peak plasma concentration at 3.63-fold and 2.68-fold, respectively, compared to mannitol alone. In contrast, no increase in mannitol concentration was shown with mannitol/AT1002 or mannitol/Pep2 compared to the control. Thus, Pep1 increased the stability or possibly reduced the instability of AT1002, which resulted in an increased permeation-enhancing effect of AT1002.

Conclusion: These results suggest the potential usefulness of C-terminal-amidated AT1002 in enhancing nasal drug delivery, which may lead to the development of a practical drug delivery technology for drugs with low bioavailability.

No MeSH data available.


Related in: MedlinePlus