The authors would like to acknowledge Jamie Thuy for technical work. This work was supported, in part, by Research grants SC CTSI NIH/NCRR/NCATS -- UL1TR000130 (D.L.D.), AA022448 (D.L.D.), and the USC School of Pharmacy. Additionally, fast dissolving oral films containing IVM were kindly provided by Cure Pharmaceutical (Oxnard, CA).

All procedure described have been approved by the Institutional Animal Care and Use Committee (IACUC) of the University of Southern California.
1. Animal Handling and Habituation
<ol>
	<li>Introduction
	<ol>
		<li>Allow mice to acclimate to the husbandry conditions and handling technique of the experiment to decrease stress during the ensuing drug administration procedures. During this time, record the body weight and food intake of each mouse; if desired, they can later be used as a non-invasive point of compar...

<ol>
	<li>Brown, A. P., Dinger, N., Levine, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stress+produced+by+gavage+administration+in+the+rat.">Stress produced by gavage administration in the rat.</a> Contemp Top Lab Anim Sci. 39 (1), 17-21 (2000).</li><li>Gonzales, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alternative+method+of+oral+administration+by+peanut+butter+pellet+formulation+results+in+target+engagement+of+BACE1+and+attenuation+of+gavage-induced+stress+responses+in+mice.">Alternative method of oral administration by peanut butter pellet formulation results in target engagement of BACE1 and attenuation of gavage-induced stress responses in mice.</a> Pharmacol Biochem Behav. 126, 28-35 (2014).</li><li>Walker, M. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+less+stressful+alternative+to+oral+gavage+for+pharmacological+and+toxicological+studies+in+mice.">A less stressful alternative to oral gavage for pharmacological and toxicological studies in mice.</a> Toxicol Appl Pharmacol. 260 (1), 65-69 (2012).</li><li>Zhang, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Voluntary+oral+administration+of+drugs+in+mice.">Voluntary oral administration of drugs in mice.</a> Protocol Exchange: Nature Publishing Group. , (2011).</li><li>Froehlich, J. C., Hausauer, B. J., Federoff, D. L., Fischer, S. M., Rasmussen, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prazosin+reduces+alcohol+drinking+throughout+prolonged+treatment+and+blocks+the+initiation+of+drinking+in+rats+selectively+bred+for+high+alcohol+intake.">Prazosin reduces alcohol drinking throughout prolonged treatment and blocks the initiation of drinking in rats selectively bred for high alcohol intake.</a> Alcohol Clin Exp Res. 37 (9), 1552-1560 (2013).</li><li>Goldkuhl, R., Carlsson, H. E., Hau, J., Abelson, K. S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+subcutaneous+injection+and+oral+voluntary+ingestion+of+buprenorphine+on+post-operative+serum+corticosterone+levels+in+male+rats.">Effect of subcutaneous injection and oral voluntary ingestion of buprenorphine on post-operative serum corticosterone levels in male rats.</a> Eur Surg Res. 41 (3), 272-278 (2008).</li><li>Marx, J. O., Vudathala, D., Murphy, L., Rankin, S., Hankenson, F. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibiotic+administration+in+the+drinking+water+of+mice.">Antibiotic administration in the drinking water of mice.</a> J Am Assoc Lab Anim Sci. 53 (3), 301-306 (2014).</li><li>Ferguson, S. A., Boctor, S. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+food+wafers+for+multiple+daily+oral+treatments+in+young+rats.">Use of food wafers for multiple daily oral treatments in young rats.</a> J Am Assoc Lab Anim Sci. 48 (3), 292-295 (2009).</li><li>Turner, P. V., Brabb, T., Pekow, C., Vasbinder, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Administration+of+substances+to+laboratory+animals:+routes+of+administration+and+factors+to+consider.">Administration of substances to laboratory animals: routes of administration and factors to consider.</a> J Am Assoc Lab Anim Sci. 50 (5), 600-613 (2011).</li><li>Bala, R., Pawar, P., Khanna, S., Arora, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orally+dissolving+strips:+A+new+approach+to+oral+drug+delivery+system.">Orally dissolving strips: A new approach to oral drug delivery system.</a> Int J Pharm Investig. 3 (2), 67-76 (2013).</li><li>Potashkin, J. A., Blume, S. R., Runkle, N. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Limitations+of+animal+models+of+Parkinson's+disease.">Limitations of animal models of Parkinson's disease.</a> Parkinsons Dis. 2011, (2010).</li><li>Machholz, E., Mulder, G., Ruiz, C., Corning, B. F., Pritchett-Corning, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Manual+restraint+and+common+compound+administration+routes+in+mice+and+rats.">Manual restraint and common compound administration routes in mice and rats.</a> Journal of Visualized Experiments JoVE. (67), e2771 (2012).</li><li>Yardley, M. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ivermectin+reduces+alcohol+intake+and+preference+in+mice.">Ivermectin reduces alcohol intake and preference in mice.</a> Neuropharmacology. 63, 190-201 (2012).</li><li>Hoggatt, A. F., Hoggatt, J., Honerlaw, M., Pelus, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+spoonful+of+sugar+helps+the+medicine+go+down:+a+novel+technique+to+improve+oral+gavage+in+mice.">A spoonful of sugar helps the medicine go down: a novel technique to improve oral gavage in mice.</a> J Am Assoc Lab Anim Sci. 49, 329-334 (2010).</li><li>Yardley, M. M., Huynh, N., Rodgers, K. E., Alkana, R. L., Davies, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oral+delivery+of+ivermectin+using+a+fast+dissolving+oral+film:+implications+for+repurposing+ivermectin+as+a+pharmacotherapy+for+alcohol+use+disorder.">Oral delivery of ivermectin using a fast dissolving oral film: implications for repurposing ivermectin as a pharmacotherapy for alcohol use disorder.</a> Alcohol. 49 (6), 553-559 (2015).</li></ol>

DD has applied for a patent as a Co-Inventor on this method and is interested in sharing this method with the scientific committee to make this technique well known and readily available for chronic animal testing (Oral Delivery of Drug Actives in Laboratory Animals Using Fast-Dissolving Oral Films (Davies, Co-Inventor, Patent Application filed 6-23-2014).

No other competing financial interests are noted.

Oral gavage is associated with numerous complications that result in potentially compromised data collection and high rates of animal attrition1. Here a simple method of oral drug delivery is introduced through which mice (and potentially other animals) can easily and reliably consume the drug of choice through an orally dissolving strip (ODS). Notably, the use of this method represents a safe, convenient, and humane alternative to oral gavage.
Other current alternatives to oral gav...

In order for the successful translation of a drug from preclinical (animal testing) to clinical development, a series of well-defined, acute and chronic pharmacological and toxicological studies need to be performed in animal models using the intended clinical route of administration (which is normally the oral route). To accomplish this goal, most pre-clinical studies currently utilize an oral gavage procedure given that this method results in a consistent and precise delivery of the test compound to the animal. However, in many instances, oral gavage is not well tolerated by the animal and accumulating evidence suggests that the method is associated with a significant amount of stress induction1.
This is especially true in the development of treatments for chronic life-long conditions that require longer-term preclinical studies, such as is the case with, but not limited to, nearly every neuropsychiatric disorder. For instance, in a transgenic mouse model of Alzheimer&#39;s disease, it was recently suggested that the gavage method itself could confound experimental results due to dosing induced stress2. Similarly, in a mouse model of alcohol consumption, unpublished findings from our own laboratory have found that the repetitive insertion of the gavage tube - in and of itself - can significantly reduce the level of alcohol intake to the point of jeopardizing the integrity of the experimental paradigm (see Figure 1).
Given the aforementioned limitations associated with the oral gavage procedure, considerable effort has been put towards developing novel methods of preclinical oral drug delivery. Current alternative approaches include incorporating the test compound into peanut butter mixtures3, gelatinous molds4-5, chocolate pellets6, drinking water7, and wafer crackers8, all of which are associated with varying degrees of issues including the inability to be used reliably in the animal model, lack of adoption of test compound uptake by the test subject or individual preference for flavor that makes the drug less readily consumed9.
The procedure introduced herein describes a method in which the desired drug is incorporated into a flavored, orally dissolving strip (ODS) that can be easily and readily be used to orally administer the test compound to the test animal. This paper focuses on demonstrating the effectiveness of the ODS method using mice. However, there is no reason to expect that the same method would not also be useful in other rodent as well as larger mammalian subjects. Clinically, the use of ODS is starting to be adopted in pediatric and geriatric patients, as a way to overcome swallowing difficulties10. We propose that ODS can also be used successfully in preclinical drug discovery programs as a safe, convenient, and humane alternative to oral gavage.

<table border="0" cellpadding="0" cellspacing="0" width="948">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:203px;">Orally Dissolving Strip&#160;</td>
			<td style="width:185px;">Cure Pharmaceutical</td>
			<td style="width:89px;"></td>
			<td style="width:471px;">Available upon special request. Alternatively, ODS may also be prepared in-house.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:203px;">Gavage Feeding Needle</td>
			<td style="width:185px;">VWR</td>
			<td align="right" style="width:89px;">7912</td>
			<td>Our rodent studies utilized a 18-gauge feeding needle. This size may need to be adjusted for use in other mammilian species.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:203px;">Sucrose</td>
			<td style="width:185px;">Sigma Aldrich</td>
			<td style="width:89px;">S0389-500G</td>
			<td></td>
		</tr>
	</tbody>
</table>

utilizing an orally dissolving strip for pharmacological and toxicological studies: a simple and humane alternative to oral gavage for animals

Prior to testing novel therapeutics in humans, short and long term preclinical (i.e., animal), repetitive pharmacological and toxicological testing is required. In most cases, the preferred route of administration is via oral delivery. At the present time, oral delivery is mostly accomplished using an oral gavage procedure, in part, because it can achieve consistent and precise dosing in the animal model. Although this method is well established it does have complications that can result in a high rate of animal attrition. To this end, the procedure introduced here describes an alternative to the oral gavage method in which the desired drug is incorporated into a tastant, orally dissolving strip (ODS) that can simply be presented to the test animal where it is then rapidly taken up with minimal manipulation of the test subject. Herein, we demonstrate that preclinical, oral drug delivery using the ODS method represents a safe, convenient, and humane alternative to oral gavage.

In the following representative investigations, social drinking was modeled using a 24 hr two-bottle choice (TBC) paradigm as previously described15. Briefly, mice had access to two bottles of solution, one of which contained water, and the other a 10% (v/v) ethanol solution. Subjects were subsequently assigned to drug treatment groups so that the average 10E (10% ethanol/90% water v/v)&#160;intake was similar across groups.
<p class="jove_content" fo:keep-together.within-p...

Watch this Scientific Journal Video about Utilizing an Orally Dissolving Strip for Pharmacological and Toxicological Studies: A Simple and Humane Alternative to Oral Gavage for Animals at JoVE.com

Utilizing an Orally Dissolving Strip for Pharmacological and Toxicological Studies: A Simple and Humane Alternative to Oral Gavage for Animals

The following protocol describes a novel method for chronic oral drug administration using an orally dissolving strip (ODS) in lieu of the more commonly used oral gavage method. We demonstrate that preclinical, oral drug delivery using the ODS method represents a safe, convenient, and humane alternative to oral gavage.

Prior to testing novel therapeutics in humans, short and long term preclinical (i.e., animal), repetitive pharmacological and toxicological testing is ...

The overall goal of this procedure is to utilize an orally dissolvable bioadhesive polymer as a safe and human means of preclinical oral drug delivery. Using this procedure, a drug of interest can be incorporated into a flavored, orally dissolving strip for the oral administration of a test compound to an experimental animal. The main advantage of this technique is that it can be utilized instances when oral gavage isn't well tolerated.To decrease stress during the ensuing drug administration procedures, allow the mice to acclimate to the husbandry conditions and handling technique of the experiment. For each mouse, determine the pre-experimental weight by grasping the mouse first by the middle of the tail, and gently lifting the animal out of the cage. Immediately place the animal into a weigh boat on an animal scale, keeping a firm grasp on the medial aspect of the tail to avoid a degloving injury and record the weight of the animal.Record the weight of the food for a recommended two consecutive days to establish the baseline food consumption of each animal during the habituation period as well. After the animals have become acclimated to their environment transfer the animals to the room in which the subsequent experiments will occur. Before beginning the experiment, autoclave sterilize two stainless steel gavage needles per group, and one additional needle for back up.Next, use a commercially available single quarter inch hole puncher to cut 0.5 centimeter diameter orally dissolving strips, or ODS, for each animal from a standard rectangular test strip. On the day of the experiment, record the weight of the animals as just demonstrated and then place the first mouse onto a metal grid cage top. Pull back on the tail with the dominant thumb, index, and third finger.The mouse will grip the cage top and stretch out its back. Grasp the scruff of the neck with the non-dominant thumb, index finger, and third finger, to comfortably restrain the animal with a secure grip, and turn the animal to an upright position. Note the color of the mucosal membranes.A pink red appearance indicates the mouse is comfortably restrained. Now submerge the bulb tip of one of the gavage needles in freshly prepared 4.25%sucrose solution to serve as a tastant adhesive to the ODS. Then press down on an ODS with the bulb tip to attach the ODS to the needle tip, and place the strip near the nostrils and mouth to allow the animal to consume the film.Alternatively, the strip can be placed directly onto the tongue for the mouse to swallow. Replace the needle mid-group during the drug dosing session to prevent the spread of infection. Here, the effects of gavage administered ivermectrine on preference drinking are shown.As observed, ivermectine significantly reduces the 10%ethanol intake across the study compared to the placebo group. Compared to the control group however, the ethanol intake values of the placebo group are much too low, and thus may not accurately model social drinking behavior. When the same experiment is performed utilizing the ODS delivery system, there is no significant difference in the ethanol intake between the control and placebo treated mice, indicating that the ODS placebo group represents a valid control with which a comparison to the ivermectine group can be made.Indeed, by these controls, ivermectine significantly and consistently reduces the 10%ethanol intake of the animals. After watching this video, you should have a good understanding of how to utilize an orally dissolving strip as a safe and humane means of oral drug delivery.

utilizing-an-orally-dissolving-strip-for-pharmacological

Research

JoVE Journal

Medicine

12.5K Görüntüleme.  University of Southern California School of Pharmacy. The overall goal of this procedure is to utilize an orally dissolvable bioadhesive polymer as a safe and human means of preclinical oral drug delivery. Using this procedure, a drug of interest can be incorporated into a flavored, orally dissolving strip for the oral administration of a test compound to an experimental animal. The main advantage of this technique is that it can be utilized instances when oral gavage isn't well tolerated.To decrease stress during the ensuing drug adminis...