Name: carotid artery infusions for pharmacokinetic and pharmacodynamic analysis of taxanes in mice
Uploaded: 2014-10-27T14:00:00
Description: Watch this Scientific Journal Video about Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice at JoVE.com

We would like to acknowledge the FCCC Laboratory Animal Facility for their support in this project. We thank Wolfe Laboratories, Inc. for their assistance in analyzing paclitaxel levels in plasma and tissue. This work was supported by National Institutes of Health grants K01CA120091 to E.H.B., and CA06927 to Fox Chase Cancer Center.

This protocol has been approved by the Fox Chase Cancer Center Institutional Animal Care and Use Committee and by the Laboratory Animal Facility, and found to be in accordance with institutional guidelines for humane treatment of animals.
1. Preliminary Preparation
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	<li>Preparation of catheter: Prepare a catheter from a short length of polyethylene tubing, modified to form a thinned, blunted end (Figure 3A). Make multiple catheters in advance and save indefinitely.
	<ol...

<ol>
	<li>Gallo, J. M., Li, S., Guo, P., Ma Reed, K., J, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Effect+of+P-Glycoprotein+on+Paclitaxel+Brain+and+Brain+Tumor+Distribution+in+Mice.">The Effect of P-Glycoprotein on Paclitaxel Brain and Brain Tumor Distribution in Mice.</a> Cancer Research. 63 (16), 5114-5117 (2003).</li><li>Sonnichen, D. S., Relling, M. 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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Physiologically+Based+Pharmacokinetic+Model+of+Docetaxel+Disposition:+from+Mouse+to+Man.">A Physiologically Based Pharmacokinetic Model of Docetaxel Disposition: from Mouse to Man.</a> Clinical Cancer Research. 13, 2768-2778 (2007).</li><li>Eiseman, J. L., 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=Plasma+Pharmacokinetics+and+Tissue+Distribution+of+Paclitaxel+in+CD2F1+Mice.">Plasma Pharmacokinetics and Tissue Distribution of Paclitaxel in CD2F1 Mice.</a> Cancer Chemotherapy and Pharmacology. 34 (6), 465-471 (1994).</li><li>Schinkel, A. 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P., 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=Reduction+of+Animal+Usage+by+Serial+Bleeding+of+Mice+for+Pharmacokinetic+Studies:+Application+of+Robotic+Sample+Preparation+and+Fast+Liquid+Chromatography+&#8211;+Mass+Spectrometry.">Reduction of Animal Usage by Serial Bleeding of Mice for Pharmacokinetic Studies: Application of Robotic Sample Preparation and Fast Liquid Chromatography &#8211; Mass Spectrometry.</a> Journal of Chromatography B: Biomedical Sciences and Applications. 754 (1), 245-251 (2001).</li><li>Berndt, K., Vogel, J., Buehler, C., Vogt, P., Born, W., Fuchs, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+method+for+repeated+drug+infusion+into+the+femoral+artery+of+mice.">A new method for repeated drug infusion into the femoral artery of mice.</a> J. Am. Assoc. 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Carotid artery infusion is a significant technique in this study of paclitaxel pharmacokinetics. Carotid artery infusion is a method to quickly distribute drug throughout the circulatory system14. The 3 hr infusion is a closer mimic of clinical administration of drugs such as taxanes than bolus injections. The surgery can be reliably performed by a single individual, surgery time is relatively short, and success rates are &#62;75%. After samples are collected, they must be analyzed by the appropriate methods. ...

Drug infusion through the carotid can be performed reliably and reproducibly by optimizing equipment and technique. The procedure is not intricate, although it does require fine control and attention to detail. Superior care and dexterity are needed to isolate the carotid artery and insert the catheter, which can generally be acquired through practice. Surgery by an experienced technician should not exceed one hour. After successful surgery, the mouse should appear normal and healthy (although the mouse may react to the actual drug infusion), and drug(s) may be administered in a controlled, uniform continuous dosage. Blood samples must be taken from a site other than the carotid artery; retro-orbital bleeds proved easy to collect and satisfactory for analysis of drug concentrations.
Catheters of optimum size and shape are an invaluable asset in performing a successful infusion11. We found the catheters available commercially often to be too large and/or too flexible to allow for convenient access to the mouse carotid artery. It proved preferable to fashion catheters from the polyethylene tubing used to connect the mouse to the infusion syringe. Thus, all the tubing, connectors and needles were of consistent dimensions, which simplified infusion assembly. Using this technique, it is not necessary to push the tip of the catheter into the artery past the point where it is still visible, and blood flow to the carotid artery is not restored until after the catheter is initially secured. This reduces the hazards of puncturing the artery or of having the catheter pushed out by the high pressure of blood flow. The catheter design herein does not incorporate a &#8220;bump&#8221; to hold it in place, so securing the catheter well with sutures and surgical tape is a priority.
Infusions may be preferable to the common i.v. bolus injections, as a better mimic of clinical administration of drugs such as taxanes3, 12, 13. The technique described here was originally developed to allow infusion into mouse models wherein access to the jugular or femoral vein was precluded by mammary tumor growth and/or excessive vascularization of the insertion area. This method may often be appropriate even in tumor-free mice: although isolating and catheterizing the carotid is slightly more invasive, we found it preferable to the jugular, because the propensity of the jugular wall to rip resulted in more failed insertions and failures to complete the 3 hr time course.
While the results shown here are from C57BL/6J (in-house-bred) mice, we have used this technique to successfully infuse paclitaxel into several strains of mice, including FVB and mixed-strains, to follow the pharmacokinetics in mouse models transgenically manipulated to down-regulate cellular transporter functions. The blood and tissue samples collected showed expected levels of paclitaxel, in the range of the levels seen after jugular infusions1. This technique may be expected to work equally well in other mouse models and with other infusion solutions.

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			<td height="25" style="height:25px;width:293px;">Table of specific reagents and equipment:</td>
			<td style="width:185px;">Source:</td>
			<td style="width:111px;">Catalog Number</td>
			<td style="width:259px;">Comments</td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">Polyethylene tubing 0.024&#8221;OD X 0.011&#8221;ID&#160;</td>
			<td style="width:185px;">Braintree Scientific, Inc.</td>
			<td style="width:111px;">PE10</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">Stainless steel port plug (30 gauge)</td>
			<td style="width:185px;">Braintree Scientific, Inc.</td>
			<td style="width:111px;">NB-30</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">2 Stainless steel connector plugs (28 gauge)</td>
			<td style="width:185px;">Braintree Scientific, Inc.</td>
			<td style="width:111px;">PP-28</td>
			<td style="width:259px;">Slightly larger than PE tubing ID, to fit snugly and keep a tight seal.</td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">3 Blunted needles (30 gauge)</td>
			<td style="width:185px;">Braintree Scientific, Inc.</td>
			<td style="width:111px;">C-30</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">3 1cc syringes</td>
			<td style="width:185px;">Becton, Dickinson and Co.</td>
			<td style="width:111px;">309659</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">Sterile 0.9% Saline solution</td>
			<td style="width:185px;">Hospira</td>
			<td style="width:111px;">0409-7984-37</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">Cath-Loc HGS Heparin/Glycerol Solution&#160;</td>
			<td style="width:185px;">Braintree Scientific, Inc.</td>
			<td style="width:111px;">HGS</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">Silk suture</td>
			<td style="width:185px;">Braintree Scientific, Inc.</td>
			<td style="width:111px;">SUT-S&#160;113</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">Vanna Scissors (micro-scissors)</td>
			<td style="width:185px;">World Precision Instruments</td>
			<td style="width:111px;">14122</td>
			<td style="width:259px;">This model has a curved tip, but straight-tip scissors work as well.</td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">Hartman Mesquito Hemostatic Forceps</td>
			<td style="width:185px;">World Precision Instruments</td>
			<td style="width:111px;">501705</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">Betadine Swabsticks</td>
			<td style="width:185px;">Perdue Products L.P.</td>
			<td style="width:111px;">BSWS1S</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">Bupivacaine</td>
			<td style="width:185px;">Hospira</td>
			<td style="width:111px;">0409-1160-01</td>
			<td style="width:259px;">May be replaced with Lidocaine, or similar local anesthesia.</td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">Paclitaxel</td>
			<td style="width:185px;">LC Laboratories</td>
			<td style="width:111px;">P-9600</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">Methanol</td>
			<td style="width:185px;">Sigma-Aldrich</td>
			<td style="width:111px;">32213</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">Micro-Hematocrit Capillary Tubes, Heparinized</td>
			<td style="width:185px;">Fisher Scientific</td>
			<td style="width:111px;">22-362-566</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">&#160;Capillary Tube Sealant&#160;</td>
			<td style="width:185px;">Fisher Scientific</td>
			<td style="width:111px;">02-678</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="70">
			<td height="70" style="height:70px;width:293px;">C57BL/6J mice</td>
			<td style="width:185px;">Fox Chase Cancer Center, Laboratory Animal Facility in-house-bred</td>
			<td style="width:111px;"></td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:293px;">API 4000 Q-Trap mass spetrometer</td>
			<td style="width:185px;">Applied Biosystems</td>
			<td style="width:111px;"></td>
			<td style="width:259px;"></td>
		</tr>
	</tbody>
</table>

carotid artery infusions for pharmacokinetic and pharmacodynamic analysis of taxanes in mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

Paclitaxel distribution follows predictable patterns during a 3 hr dosing regimen of a 15 min high-speed infusion, followed by a 165 min low-speed infusion.
Figure 1 shows a comparison of jugular vein-infusion plasma paclitaxel concentrations and carotid artery-infusions. The paclitaxel concentrations drop quickly in the first 15 min following an initial high volume infusion, and then level off over the next 150 min. By comparison, paclitaxel levels in a poor infusion start of...

Watch this Scientific Journal Video about Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice at JoVE.com

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

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