J. vet. Pharmacol. Therap. 27, 369–372, 2004.
SHORT COMMUNICATION
Sedative effects and serum drug concentrations of oxymorphone and
metabolites after subcutaneous administration of a liposome-encapsulated
formulation in dogs
L. J. SMITH*
L. KRUGNER-HIGBY  
L. A. TREPANIER à
D. E. FLASKA*
V. JOERS   &
T. D. HEATH §
*Department of Surgical Sciences,  Research Animal Resource Center, àDepartment of Medical Sciences, School of Veterinary Medicine and
§
School of Pharmacy, University of Wisconsin, Madison, WI, USA
(Paper received 19 November 2003; accepted for publication 11 May 2004)
Lesley J. Smith, Department of Surgical Sciences, School of Veterinary Medicine, University of Wisconsin, 2015 Linden Drive,
Madison, WI 53706, USA. E-mail: smithl@svm.vetmed.wisc.edu
Adequate treatment of chronic pain is increasingly recognized as
fundamental to quality companion animal medicine. Companion
animals, particularly dogs, can be expected to have an increased
lifespan because of modern improvements in veterinary preven-
tative care. With age often come painful chronic conditions such
as osteoarthritis and neoplastic disease. The ideal analgesic for
treatment of chronic pain would be effective, free from significant
side effects, and have a dosage interval and route of adminis-
tration that is convenient to the caretaker.
Encapsulation of drugs into liposomes is one approach to
providing a sustained-release drug delivery formulation. Hydro-
philic/lipophobic drugs such as morphine have been successfully
encapsulated using a multilamellar vesicular system called
DepofoamÒ (SkyePharma Inc., San Diego, CA, USA) (Howell,
2001). Oxymorphone is a hydrophobic, lipophilic opioid that is
not well suited to encapsulation using multilamellar vesicular
technology. We have developed a novel technique for liposome-
encapsulation of oxymorphone (Krugner-Higby et al., 2002).
This formulation of liposome-encapsulated (LE) oxymorphone
releases drug slowly into the systemic circulation from a
subcutaneous depot. Oxymorphone is a common, safe, and
extremely effective analgesic that is metabolized by the liver with
renal and biliary excretion of metabolites (Cone et al., 1983). It is
generally well tolerated by a wide patient population. Side effects
of oxymorphone in dogs are usually mild at clinically relevant
doses, but can include dose-dependent sedation, respiratory
depression, panting, bradycardia, nausea, urinary retention, and
constipation (Pascoe, 2000a,b; Smith et al., 2001).
The objective of the present study was to assess the sedative
and physiologic changes associated with subcutaneous admin-
istration of LE oxymorphone in healthy dogs and to compare the
serum concentration vs. time profiles of LE and standard (STD)
oxymorphone.
Ó 2004 Blackwell Publishing Ltd
Twenty-five adult dogs, including beagles (n ¼ 14; 12
females, two males) and hounds (n ¼ 11; nine females, two
males) aged 1–5 years old (median age 1.5 years) and weighing
between 6.9 and 35.7 kg (median weight ¼ 9.46 kg) were used
in this study. Normal health status and organ function were
confirmed by physical exam and a complete blood count and
serum chemistry profile prior to inclusion in the study. None of
the animals received any medications in the 2 weeks prior to
study. Animals were housed individually (hounds) or in groups
of 2–4 (beagles), and were fed a commercial diet (Harlan Teklad
21% Lab Dog Diet (W); Harlan Laboratories, Madison, WI, USA)
and water ad libitum throughout the study. The protocol for this
study was approved by the University of Wisconsin, School of
Veterinary Medicine Animal Care and Use Committee.
Standard oxymorphone was purchased from a commercial
source (NumorphanÒ Endo Laboratories, Chadds Ford, PA,
USA). LE oxymorphone was synthesized using a novel technique
previously reported by this laboratory (Smith et al., 2003).
Briefly, dehydration–rehydration vesicles containing oxymor-
phone were prepared by the method of Kirby and Gregoriadis
(1984). Oxymorphone in the liposome preparations was quant-
itated as previously described (Smith et al., 2003).
Two doses of STD oxymorphone and LE oxymorphone were
investigated. Doses of LE oxymorphone chosen for this study
were 0.5 and 1.0 mg/kg, while generally recommended doses of
STD oxymorphone are 0.05–0.1 mg/kg for analgesia in dogs
(Pascoe, 2000a,b). The proposed equipotent doses of liposomal
opioid were designed to be 10 times greater than the commonly
recommended doses of the standard formulation, based on
previous experience with this formulation (Krugner-Higby et al.,
2003; Smith et al., 2003; R. Willis, personal communications),
and previous studies using a similar, morphine-based LE
preparation in dogs that have employed doses that were
369
370 L. J. Smith et al.
generally ten times greater than those for standard drug
preparations of that opioid (Yaksh et al., 1999, 2000). One dose
of the liposomal vehicle (unloaded liposomes) was also investi-
gated to confirm the absence of pharmacodynamic properties of
the vehicle. Each dog received only one treatment and treat-
ments were administered subcutaneously (s.c.). Treatment
groups were as follows: group 1, 1.0 mg/kg LE oxymorphone
(n ¼ 6); group 2, 0.5 mg/kg LE oxymorphone (n ¼ 5); group 3,
0.1 mg/kg STD oxymorphone (n ¼ 6); group 4, 0.05 mg/kg
STD oxymorphone (n ¼ 6); group 5, 0.7 mL of liposomal vehicle
(n ¼ 2).
In groups 1 and 2 (LE oxymorphone groups), blood was
collected before and 0.5, 1, 2, 4, 8, 12, 16, and 24 h after drug
administration, and then daily for 5 days at the same time of day
as treatment administration. In groups 3, 4, and 5 (STD
oxymorphone and liposomal vehicle groups), blood was collected
before and 0.5, 1, 2, 4, 8, 24, and 48 h after drug administra-
tion. Blood samples were separated by centrifugation and serum
was stored at )70 °C for <1 month until analysis.
A sedation score was recorded for each dog prior to drug
administration (baseline) and immediately prior to each venous
blood collection (Smith et al., 2001). Changes in sedation score,
heart rate, respiratory rate, and temperature were analyzed
using a Wilcoxon Rank Sum test for nonparametric data with
P < 0.05 considered significant.
Drug concentrations in serum were quantitated using a
commercial ELISA (Neogen Inc., Lexington, KY, USA), which
measures both parent drug and the glucuronide metabolites of
oxymorphone. The assay yielded an average intra-assay variab-
ility of 2.83% and inter-assay variability of 7.61% across the
range of expected drug concentrations, i.e. from 0 to 100 ng/mL.
The lower limit of quantitation was approximately 1.5 ng/mL.
Comparison of the mean serum concentrations achieved at each
common time point between groups was performed using ANOVA
with post hoc testing using the method of Tukey (SigmaStat,
version 2.03, SPSS Science, Chicago, IL, USA).
Local skin reactions or obvious infection as a result of injection
of the standard or liposomal preparations of drug were not
observed in any dog. Injection of the liposomal vehicle had no
effect on heart rate, respiratory rate, temperature, or sedation
score, and no oxymorphone or metabolite was detected in serum
of dogs that received the liposomal vehicle.
Figure 1 shows sedation scores in dogs that received LE
oxymorphone or STD oxymorphone at the lower and higher
doses. Sedation score peaked at 30 min in dogs that received STD
oxymorphone, and at 1 h in dogs that received the LE
oxymorphone. Sedation score was significantly higher at 1 h
in dogs that received 1.0 mg/kg of LE oxymorphone compared
with dogs that received either dose of the standard preparation.
By 2 h, there was no difference in sedation score between
groups.
As expected, a mild increase in respiratory rate, manifested as
panting, was observed in some dogs, but was not significantly
different from baseline for any group. Baseline respiratory rate
for all groups was 29.6 ± 2.2 breaths per minute (mean ± SD).
At t ¼ 1 h, respiratory rates were: 30.6 ± 18.4 (group 1:
Ó 2004 Blackwell Publishing Ltd, J. vet. Pharmacol. Therap. 27, 369–372
7.5
1 mg/kg LE
0.5mg/kg LE
0.1 mg/kg STD
Sedation score
5.0 0.05 mg/kg STD
2.5
0.0
Baseline 30 min 1h 2h 4h 8h 12h
Fig. 1. Sedation scores in dogs that received either standard (STD)
oxymorphone or liposome-encapsulated (LE) oxymorphone. *Significantly
higher than the two doses of STD oxymorphone at that time point.
1.0 mg/kg LE oxymorphone); 24.0 ± 2.8 (group 2: 0.5 mg/kg
LE oxymorphone); 30.0 ± 16.9 (group 3: 0.1 mg/kg STD
oxymorphone); 26.6 ± 2.2 (group 4: 0.05 mg/kg STD oxymor-
phone); and 26.0 ± 2.8 (group 5: liposomal vehicle). Without
access to blood gas analysis, it is not possible to determine
whether the observed changes in respiratory rate correlated with
significant changes in arterial pH, PaO2 or PaCO2.
Mean rectal temperature tended to decrease, although not
significantly, from a baseline value (averaged across all groups) of
101.3 ± 1.9–99.7 ± 1.3 °F at t ¼ 1 h in all groups that received
some form of oxymorphone. This reduction in temperature may
be attributable to the increase in respiratory rate, or panting, in
some dogs. Figure 2 shows the changes in heart rate that
occurred after administration of the LE oxymorphone or STD
oxymorphone. A significant decrease in heart rate was observed
from baseline to 2 h after all doses of LE or STD oxymorphone
were administered. The lowest heart rate observed (64 bpm) was
at t ¼ 1 h in one dog that received 1.0 mg/kg of LE oxymor-
phone. In all groups that received any form of oxymorphone,
actual heart rates were never less than what would be considered
to be within a clinically acceptable range for healthy dogs.
Figure 3 shows serum concentrations of oxymorphone and its
glucuronide in all four groups of dogs. Serum concentrations
were significantly higher in the dogs that received 1.0 mg/kg of
LE oxymorphone at all time points measured. By 24 h,
concentrations of drug in dogs that received either dose of STD
oxymorphone or the lower dose of LE oxymorphone fell below
the limit of quantitation (1.5 ng/mL). The higher dose of LE
oxymorphone therefore resulted in more persistent serum
concentrations of both parent drug and metabolites than either
the lower dose of LE drug or either dose of the standard
preparation.
Currently available opioid analgesics that can be prescribed for
Ôat homeÕ use in the US and that provide an extended duration of
effect include the fentanyl patch, and oral formulations of
butorphanol, morphine, oxycodone, oxycontin, and codeine. All
of these opioid formulations present a significant risk of drug
diversion by human caretakers (Purucker & Swann, 2000;
 Liposome-encapsulated oxymorphone in dogs 371

1 mg/kg LE
0.5 mg/kg LE
0.1mg/kg STD
150 0.05mg/kg STD

Heart race (beats/min)

100

Fig. 2. Changes in heart rate in dogs that

50
received either standard (STD) oxymorphone
or liposome-encapsulated (LE) oxymorphone.
*Significantly different than baseline across
groups; **significantly different from other 0
groups at that time point. baseline 30min 1h 2h 4h 8h 12h 16h day 1 day 2 day 3 day 4 day 5

75
1 mg/kg LE
* *
0.5 mg/kg LE
metabolites] (ng/mL)
0.1mg/kg STD
*
[oxymorphone &

50 0.05mg/kg STD
Total serum

*
* Presumed
Fig. 3. Serum oxymorphone and metabolite 25 * * * therapeutic
range
concentrations in all groups of dogs from
baseline to 5 days postadministration of
* *
oxymorphone. *Significantly different than 0
* *
the lower doses of STD and LE oxymorphone. baseline 30 min 1 h 2h 4h 8h 12 h 16 h day 1 day 2 day 3 day 4 day 5

Passik, 2001; Wasserman, 2001) or accidental access to
children (Hardwick et al., 1997). In addition, bioavailability
and efficacy of these opioid formulations are variable in humans
(Gourlay et al., 1997) and dogs (Kyles et al., 1996; Dohoo, 1997;
Egger et al., 1998). There is a need for a long acting,
physiologically safe, and effective analgesic for dogs that can
be administered without the risk of illicit access by caretakers.
The purpose of the current study was to assess the sedative
effects and the serum concentration vs. time behavior of a single
subcutaneous injection of LE oxymorphone, and to compare
these results to those measured after subcutaneous injection of
an equipotent dose of the standard, aqueous formulation of
oxymorphone in dogs.
Our data indicate that administration of 1 mg/kg of LE
oxymorphone to healthy dogs resulted in moderate, transient
bradycardia and significant, but clinically acceptable, sedation at
1 h after subcutaneous administration. The significant sedation
observed at 1 h after administration of the higher dose of LE
oxymorphone correlates with the higher serum drug concentra-
tions seen with this dosing regimen. The lack of a significant
difference in sedation at the 30-min assessment point, despite
serum oxymorphone concentrations that were comparable with
the 1 h time point, may have been due to an effect lag as
oxymorphone diffused to the CNS. Sedation scores were similar
in all dogs at 2 h after administration of either 1.0 mg/kg LE
oxymorphone, or one-tenth that dose (0.1 mg/kg) of STD
oxymorphone. Of note is the lack of significant differences in
sedation at later time points, despite a consistently higher
oxymorphone concentration throughout the sampling period.
This may reflect the presence of oxymorphone glucuronide,
which may have less sedative effects than the parent drug.
Using an ELISA assay that measures both oxymorphone and
its glucuronide metabolite, we observed serum concentrations
after administration of 0.1 mg/kg STD oxymorphone of
19.1 ± 11.2 and 11.1 ± 3.6 ng/mL (mean ± SD) at 2 and
4 h, respectively (see Fig. 3). There are no published studies that
have correlated serum concentrations of oxymorphone with
clinical analgesia in dogs. Current dosing recommendations for
STD oxymorphone in dogs, however, are 0.05–0.1 mg/kg
administered every 3–4 h (Pascoe, 2000a,b). This would imply
that serum concentrations of oxymorphone are maintained
within an apparent therapeutic range for up to 4 h after
administration. From these clinical practices, along with our
data in the current study, we would infer that serum concen-
trations of total oxymorphone (parent drug and metabolites) less
than 10 ng/mL are below the therapeutic range. After admini-
stration of what we estimated was an equipotent dose of LE
oxymorphone (1.0 mg/kg), we observed serum concentrations of
drug >10 ng/mL for over 2 days after a single injection.
It is important to note that because our assay measures both
parent drug and glucuronide metabolites, we cannot state that
measured total drug concentrations correspond directly to
pharmacologic activity. Without concurrent algesiometry, or a
more specific assay, such as HPLC, we cannot equate the

Ó 2004 Blackwell Publishing Ltd, J. vet. Pharmacol. Therap. 27, 369–372

372 L. J. Smith et al.
reported serum concentrations with clinical effectiveness. It will
be important to follow-up on these studies with analgesic testing
using a variety of experimental stimuli in dogs treated with
liposomal oxymorphone. We are also developing a more specific
HPLC assay to clarify the relationship between serum drug
concentrations and pharmacodynamic effects.
The results of this study indicate that the subcutaneous
administration of 1.0 mg/kg LE oxymorphone is not associated
with excessive sedation in healthy dogs, and results in total
serum drug concentrations >10 ng/mL for at least 2 days
after a single dose. These concentrations are comparable with
those sustained for only 4 h after a single dose of STD
oxymorphone. More specific pharmacokinetic and pharmaco-
dynamic studies are warranted to test this liposomal formu-
lation of oxymorphone for analgesic efficacy, and for safety in
geriatric animals or those with systemic disease. The results of
this preliminary study have significant potential implications
for pain management in companion animal and human
medicine. Liposome-encapsulation of oxymorphone provides a
formulation of drug that can potentially be administered at
delayed intervals and that may induce fewer side effects than
standard formulations of opioids due to the avoidance of bolus
absorption to therapeutic concentrations. Clearly, more studies
in companion animals, and, ultimately, humans are warranted
to define the pharmacokinetic and analgesic properties of LE
oxymorphone.
ACKNOWLEDGMENTS
Funded by the Companion Animal Foundation, University of
Wisconsin, School of Veterinary Medicine, and the American
College of Laboratory Animal Medicine. The authors wish to
thank Dr Barb Gilligan and Dr Jim Southard of the Transplant
Research Laboratory, University of Wisconsin Medical School
and Dr Mark Markel and the Comparative Orthopedic Research
Laboratory, University of Wisconsin School of Veterinary
Medicine for their generous provision of dogs for this study,
and Claudia Hirsch and the animal care staff at Charmany
Instructional Facility for their care of animals.
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