Epithelial Transport and Bioavailability of Intranasally Administered Human Growth Hormone Formulated with the Absorption Enhancers Didecanoy‐L‐α‐phosphatidylcholine and α‐Cyclodextrin in Rabbits

doi:10.1002/jps.2600831212

Journal of Pharmaceutical Sciences

Volume 83, Issue 12, December 1994, Pages 1706-1711

https://doi.org/10.1002/jps.2600831212 Get rights and content

Abstract

The transepithelial transport of biosynthetic human growth hormone (hGH) formulated with the absorption enhancers didecanoyl‐L‐α‐phosphatidylcholine (DDPC) and α‐cyclodextrin (α‐CD) was studied after intranasal administration to rabbits. Plasma concentrations of the hormone were determined until 240 min post administration by ELISA, and the absolute bioavailability was estimated to be in the vicinity of 20%. The localization of hGH was studied 15 min after application of the powder formulation in the initial absorptive phase. To visualize the hormone, a two‐step indirect immuno‐gold technique was used on semithin and ultrathin cryosections and Epon sections. Polyclonal rabbit anti‐hGH was used as primary antibody and gold‐conjungated goat anti‐rabbit IgG as secondary antibody, succeeded by silver enhancement. Growth hormone was mainly found in the cytoplasm and nuclei of ciliated cells, showing distinct morphological signs of early necrosis, and in lamina propria, including the venules. Minute amounts of hGH were found in endocytotic vesicles in morphologically normal epithelial cells and in the intercellular compartment. We conclude that the major transport route of hGH formulated with absorption enhancers DDPC and α‐CD was transcellular through lethally damaged ciliated cells.

References (29)

K. Albertsson‐Wikland et al.
Acta Pediatr. Scand.
(1986)
L.M. Winer et al.
J. Clin. Endocrinol. Metab.
(1990)
R.G. Clark et al.
J. Endocr.
(1985)
C. McMartin et al.
J. Pharm. Sci.
(1987)
A.L. Daugherty et al.
Int. J. Pharm.
(1988)
V.H.L. Lee et al.
Crit. Rev. Ther. Drug Carr. Syst.
(1991)
B. Dinesen et al.
Anal. Chim. Acta
(1984)
K.T. Tokuyasu
J. Cell Biol.
(1973)
K.T. Tokuyasu et al.
J. Cell Biol.
(1976)
L. Scopsi et al.
Histochemistry
(1986)

L. Bastholm et al.

J. Electron Microsc. Tech.

(1986)

G. Griffiths et al.

J. Ultrastruc. Res.

(1984)

D.P. Knight

D.O. Slauson et al.

Cited by (31)

Human growth hormone: New delivery systems, alternative routes of administration, and their pharmacological relevance
2011, European Journal of Pharmaceutics and Biopharmaceutics
Citation Excerpt :
In summary, surfactant materials and bile salts have been shown to enhance GH bioavailability. However, the enhancing effect is usually correlated to the damage caused to the nasal membrane; although for other enhancers such as cyclodextrins, chitosan and selected phospholipids (e.g., LCP), the absorption enhancing effect is reported to outweigh irritation to the mucosa [49,50,52]. The possibility of delivering therapeutic amounts of GH by the intranasal route will depend on the quantities that can be dosed in a patient-friendly system and the potential nasal and systemic toxicity of the selected enhancer material.
The availability of recombinant human growth hormone (GH) has broadened its range of clinical applications. Approved indications for GH therapy include treatment of growth hormone deficiency (in children and in adults), Turner syndrome, Prader–Willi syndrome, chronic renal insufficiency and more recently, idiopathic short stature in children, AIDS-related wasting and fat accumulation associated with lipodystrophy in adults. Therapy with GH usually begins at a low dose and is gradually titrated to obtain optimal efficacy while minimizing side effects. It is usually administered on a daily basis by subcutaneous injection, since this was considered to impact upon patient compliance, extended-release GH preparations were developed and new delivery platforms – e.g., auto-injectors and needle-free devices – were introduced in order to improve not only compliance and convenience but also dosing accuracy. In addition, alternative less invasive modes of administration such as the nasal, pulmonary and transdermal routes have also been investigated. Here, we provide an overview of the different technologies and routes of GH administration and discuss the principles, limitations and pharmacological profiles for each approach.
Reestablishment of the nasal permeability barrier to several peptides following exposure to the absorption enhancer tetradecyl-β-D-maltoside
2010, Journal of Pharmaceutical Sciences
Citation Excerpt :
Therefore, agents known as absorption or permeation enhancers have been added to nasal formulations containing peptides to facilitate their absorption into the systemic circulation.5–7 Successful peptide drug absorption from the nasal route has been achieved with several reagents, including chitosan, didecanoyl-L-alpha-phosphatidylcholine, dimethyl-β-cyclodextrin, and the alkylglycosides.8–11 The alkylglycosides are a family of nonionic surfactants, consisting of alkyl chains of various lengths (6–16 carbon units in length) linked to a monosaccharide (6–12 carbon unit chains) or disaccharide (10–16 carbon unit chains) moiety.12
Regular insulin, NPH insulin, glargine insulin, calcitonin, and human growth hormone were administered to rats nasally with 0.125% tetradecyl-β-D-maltoside (TDM), or at various times after TDM treatment. Absorption of all five peptides was enhanced initially and diminished in a time-dependent manner as the interval between administration of TDM and the peptide increased. Changes in nasal morphology were also assessed via transmission electron microscopy (TEM) immediately after TDM treatment and at various times thereafter. TEM analysis demonstrated that 0.125% TDM caused a rapid and transient alteration in the morphology of the apical membrane surface. Fewer cilia were observed and cell–cell junctions were difficult to visualize, but no epithelial cell erosion was apparent. Two hours after TDM treatment, the apical membrane surface once again contained abundant cilia and cell–cell junctions were readily visualized. The complete recovery of the nasal permeability barrier to several different peptides following TDM administration and the concomitant histological evidence demonstrate that TDM treatment transiently perturbs the nasal mucosa to stimulate peptide drug absorption and does not produce irreversible damage to the cells that line the nasal cavity. © 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 99: 1912–1920, 2010
Nasal absorption of metoclopramide from different Carbopol<sup>®</sup> 981 based formulations: In vitro, ex vivo and in vivo evaluation
2006, European Journal of Pharmaceutics and Biopharmaceutics
There is a need for nasal drug delivery of metoclopramide HCI (MTC) in specific patient populations where the use of commercially available intravenous and oral dosage forms may be inconvenient and/or unfeasible. In this perspective, nasal dosage forms (solution, gel and lyophilized powder) of MTC were prepared by using a mucoadhesive polymer Carbopol 981 (CRB 981). The drug release studies of formulations were performed by using a modified horizontal diffusion chamber with cellulose membrane and excised cattle nasal mucosa as diffusion barriers. After the ex vivo experiments, the morphological appearances of the nasal mucosa were analyzed with the light microscopic studies. In vivo experiments were carried on sheep model. The release of MTC from solution and powder formulations was found higher than gel formulation (p < 0.05) and no severe damage was found on the integrity of nasal mucosa after ex vivo experiments. The penetration enhancing effect of dimethyl-β-cyclodextrin (DM-β-CD) used in powder formulations was observed in ex vivo and in vivo experiments. In contrast to in vitro and ex vivo experiments the nasal bioavailability of gel formulation was found higher than those of the solution and powder (p < 0.05) and might represent a promising novel tool for the systemic delivery of MTC.
Intranasal delivery of recombinant human growth hormone (somatropin) in sheep using chitosan-based powder formulations
2005, European Journal of Pharmaceutical Sciences
The effectiveness of chitosan in promoting the intranasal bioavailability of recombinant human growth hormone (hGH) has been evaluated. hGH was formulated with chitosan to produce a powder blend (Formulation A) and granules (Formulation B) for intranasal administration. The in vivo pharmacokinetic performance of the formulations was evaluated in a group of six sheep in a randomised crossover study. A subcutaneous injection of hGH solution was administered as a control. The intranasal and subcutaneous doses of hGH were 0.3 and 0.03 mg/kg, respectively. The intranasal formulations appeared to be well tolerated. Mean bioavailabilities of hGH from Formulations A and B were 14 and 15%, respectively relative to subcutaneous injection. It is concluded that chitosan-based intranasal powder formulations may provide a practical means for non-injectable delivery of hGH and, potentially, other therapeutic protein molecules.
Correlation of tetradecylmaltoside induced increases in nasal peptide drug delivery with morphological changes in nasal epithelial cells
2004, Journal of Pharmaceutical Sciences
Citation Excerpt :
After cooling, blocks were cut into regular or thin sections and post-stained with toludine blue for LM or uranyl acetate and lead citrate for electron microscopy and viewed on a light microscope (Nikon labophot-2, Melville, NY) or two transmission electron microscopes (model 1200 EX II and JEM-100 CX TEMSCAN microscope, Tokyo, Japan). Samples were evaluated by three investigators for signs of morphologic alterations as described previously (i.e., cellular rearrangement or denudement, pyknotic nuclei, and cilia alterations).17 These evaluations were done in order to assess the level of acute toxicity which might be expected in tissue exposed to TDM.
The effect of tetradecylmaltoside (TDM) on nasal peptide drug absorption was assessed with four peptides of distinct molecular size: insulin (5.7 kDa), leptin (16 kDa), somatropin (22.1 kDa), and epoetin alfa (30.4 kDa). The nasal uptake of the smallest peptides, insulin and leptin, was significantly increased at a TDM concentration of only 0.06%. The uptake of somatropin was significantly increased when concentrations of 0.125% or more were used. The uptake of the largest peptide, epoetin alfa, was not significantly increased, in the presence of 0.125−0.5% TDM. Light microscopy revealed that formulations containing 0.125% TDM caused moderate alterations in nasal epithelial cell morphology, while higher concentrations of TDM (0.5%), caused more extensive morphological changes. Following treatment with 0.125% TDM, the distribution of cilia was altered and the number of pinocytotic vesicles was increased, at a time that correlated with increased nasal absorption of insulin. Consistent with these findings, FITC-insulin applied nasally in the absence of TDM did not enter nasal epithelial cells, whereas FITC-insulin co-administered with 0.125% TDM was internalized into the cells, with a uniform distribution, consistent with transcellular movement of the peptide through the cells.
Thiomers in noninvasive polypeptide delivery: In vitro and in vivo characterization of a polycarbophil-cysteine/glutathione gel formulation for human growth hormone
2004, Journal of Pharmaceutical Sciences
Citation Excerpt :
Because nasal permeability decreases rapidly for hydrophilic molecules >1 kDa, the absorption of polypeptides such as hGH with a molecular mass of 22 kDa is negligible. To improve the nasal absorption of large‐size molecules, permeation enhancers must be included in drug formulations.23 In first orientating permeation studies, the potential of the PCP‐Cys/GSH system as an alternative nasal permeation enhancer was evaluated using FD‐4, because it is a well‐characterized paracellular marker and exhibits the advantageous property of not being enzymatically degraded in comparison to polypeptides.
This study was aimed at investigating the potential of a new polycarbophil‐cysteine (PCP‐Cys)/glutathione (GSH) gel formulation to enhance the permeation of the model drug human growth hormone (hGH) across nasal mucosa in vitro and in vivo. The aqueous nasal gel contained PCP‐Cys, GSH, and hGH in a final concentration of 0.3%, 0.5%, and 0.6% (m/v), respectively. In vitro permeation studies were performed in Ussing chambers on freshly excised bovine nasal mucosa using fluorescence‐labeled dextran (molecular mass: 4.3 kDa; FD‐4) and hGH (FITC‐hGH). The release profile of FITC‐hGH from the gel formulation and an unmodified PCP control formulation was determined. Furthermore, in vivo studies in rats were performed comparing the PCP‐Cys/GSH/hGH gel with PCP/hGH control gel and physiological saline. The permeation of FD‐4 and FITC‐hGH across the nasal mucosa was improved two‐fold and three‐fold, respectively, in the presence of PCP‐Cys/GSH. The PCP‐Cys/GSH/hGH gel and the PCP/hGH control gel showed the same biphasic and matrix‐controlled drug release. The nasal administration of the PCP‐Cys/GSH/hGH gel formulation to rats resulted in a significantly increased and prolonged hGH plasma concentration–time profile versus unmodified PCP gel and physiological saline. According to these results, PCP‐Cys gels might represent a promising new strategy for systemic nasal polypeptide delivery. © 2004 Wiley‐Liss, Inc. and the American Pharmacists Association J Pharm Sci 93:1682–1691, 2004

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Research ArticlesEpithelial Transport and Bioavailability of Intranasally Administered Human Growth Hormone Formulated with the Absorption Enhancers Didecanoy‐L‐α‐phosphatidylcholine and α‐Cyclodextrin in Rabbits

Abstract

Acta Pediatr. Scand.

J. Clin. Endocrinol. Metab.

J. Endocr.

J. Pharm. Sci.

Int. J. Pharm.

Crit. Rev. Ther. Drug Carr. Syst.

Anal. Chim. Acta

J. Cell Biol.

J. Cell Biol.

Histochemistry

J. Electron Microsc. Tech.

J. Ultrastruc. Res.

Research Articles
Epithelial Transport and Bioavailability of Intranasally Administered Human Growth Hormone Formulated with the Absorption Enhancers Didecanoy‐L‐α‐phosphatidylcholine and α‐Cyclodextrin in Rabbits