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Medicine

Novel Methods for Intranasal Administration Under Inhalation Anesthesia to Evaluate Nose-to-Brain Drug Delivery

Published: November 14th, 2018

DOI:

10.3791/58485

1Laboratory of Pharmaceutics, School of Pharmacy, Nihon University
* These authors contributed equally

Here, we describe two novel methods of stable intranasal administration under inhalation anesthesia with minimal physical stress for experimental animals. We also describe a method for quantitative evaluation of drug distribution levels in the brain via the nose-to-brain pathway using radiolabeled [14C]-inulin as a model substrate of water-soluble macromolecules.

Intranasal administration has been reported to be a potential pathway for nose-to-brain delivery of therapeutic agents that circumvents the blood-brain barrier. However, there have been few reports regarding not only the quantitative analysis but also optimal administration conditions and dosing regimens for investigations of nose-to-brain delivery. The limited progress in research on nose-to-brain pathway mechanisms using rodents represents a significant impediment in terms of designing nose-to-brain delivery systems for candidate drugs.

To gain some headway in this regard, we developed and evaluated two novel methods of stable intranasal administration under inhalation anesthesia for experimental animals. We also describe a method for the evaluation of drug distribution levels in the brain via the nose-to-brain pathway using radio-labeled [14C]-inulin (molecular weight: 5,000) as a model substrate of water-soluble macromolecules.

Initially, we developed a pipette-based intranasal administration protocol using temporarily openable masks, which enabled us to perform reliable administration to animals under stable anesthesia. Using this system, [14C]-inulin could be delivered to the brain with little experimental error.

We subsequently developed an intranasal administration protocol entailing reverse cannulation from the airway side through the esophagus, which was developed to minimize the effects of mucociliary clearance (MC). This technique led to significantly higher levels of [14C]-inulin, which was quantitatively detected in the olfactory bulb, cerebrum, and medulla oblongata, than the pipette method. This appears to be because retention of the drug solution in the nasal cavity was substantially increased by active administration using a syringe pump in a direction opposite to the MC into the nasal cavity.

In conclusion, the two methods of intranasal administration developed in this study can be expected to be extremely useful techniques for evaluating pharmacokinetics in rodents. The reverse cannulation method, in particular, could be useful for evaluating the full potential of nose-to-brain delivery of drug candidates.

Biomedicines such as peptides, oligonucleotides, and antibodies are considered to have potential application as novel therapeutic agents for refractory central nervous system disorders that currently have no curative therapy. However, because most biomedicines are water-soluble macromolecules, delivery from the blood into the brain via intravenous or oral administration is extremely difficult due to impedance of the blood-brain barrier (BBB).

In recent years, intranasal administration has been reported to be a potential pathway for nose-to-brain delivery of therapeutic agents that avoids the BBB1,

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This animal study (#AP17P004) was performed in accordance with the guidelines approved by the Nihon University Animal Care and Use Committee (Tokyo, Japan). This study (#17-0001) was approved by the Radioisotope Center of the School of Pharmacy, Nihon University.

1. Animals Used for Intranasal Administration Under Inhalation Anesthesia

  1. House the experimental mice in stainless-steel cages under a 12-h light/dark cycle (light on 8:00 AM–8:00 PM), with a controlled temperature m.......

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Figure 3 shows the [14C]-inulin levels (ID%⁄g brain) in the olfactory bulb (A), cerebrum (B), and medulla oblongata (C) obtained using the two types of intranasal administration assessed in the present study. Intranasal administration using the pipette method enabled delivery of [14C]-inulin into the brain using openable inhalation masks (Figure 1). Under inhalation anesthesia, the quantitative results.......

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The nose-to-brain delivery of drugs is expected to have a pronounced effect on central nervous system disorders because this pathway represents a direct transportation route that bypasses the BBB. Three different nose-to-brain pathways have been reported to date8. The first is the olfactory nerve pathway, which passes from the olfactory mucosa in the nasal mucosa to the forebrain via the olfactory nerve. The second is the trigeminal nerve pathway, which passes from the respiratory mucosa in the na.......

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This study was supported in part by the Private University Research Branding Project from MEXT; a Grant-in-Aid for Scientific Research (C) (17K08249 [to T.K. and T.S.]) from the Japan Society for the Promotion of Science (JSPS); a grant for cooperative research from the Hamaguchi Foundation for the Advancement of Biochemistry [to T.S.], and the Takeda Science Foundation [to T.K.]. We thank Mr. Yuya Nito and Ms. Akiko Asami for their valuable technical assistance in conducting the experiments.

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Name Company Catalog Number Comments
ddY mouse Japan SLC, Inc. Male, 4-6 weeks, 20-30 g
Isoflurane Pfizer v002139
Isoflurane setup SHINANO manufacturing CO. LTD. SN-487-OTAir, SN-489-4
Isoflurane mask SHINANO manufacturing CO. LTD. For small rodents
Isoflurane mask (Opneable type) SHINANO manufacturing CO. LTD. Special orders
Anesthesia Box SHINANO manufacturing CO. LTD. SN-487-85-02
Animal experiments scissors-1 NATSUME SEISAKUSHO CO., LTD. B-27H
Animal experiments scissors-2 NATSUME SEISAKUSHO CO., LTD. B-13H
Tweezers-1 FINE SCIENCE TOOLS Inc. 11272-30 Dumont #7 Dumoxel
Tweezers-2 NATSUME SEISAKUSHO CO., LTD. A-12-1
Cannula tube (PE-50) Becton, Dickinson and Company. 5069773 I.D.: 0.58 mm, O.D.: 0.965 mm
Cannula tube (SP-10) NATSUME SEISAKUSHO CO., LTD. KN-392 I.D.: 0.28 mm, O.D.: 0.61 mm
Shaver MARUKAN, LTD. DC-381
Stereoscopic microscope Olympus Corporation SZ61
Needle 27G 1/2 in 13 mm TERUMO CORPORATION NN-2738R
1 mL syringe TERUMO CORPORATION SS-01T
Syringe pump Neuro science NE-1000
Cellulose membrane Toyo Roshi Kaisya, Ltd. 00011090
Micro spatula Shimizu Akira Inc. 91-0088
Micropipette (0.5-10 uL) Eppendorf AG Z368083
Pipette chip Eppendorf AG 0030 000.811
Tape TimeMed Labeling System, Inc. T-534-R For fixing mouse
[14C]-Inulin American Radiolabeled Chemicals Inc. ARC0124A 0.1 mCi/mL
EtOH Wako Pure Chemical Industries, Ltd. 054-00461
Liquid scintillation counter Perkin Elmer Life and Analytical Sciences, Inc Tri-Carb 4810TR

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