TYPE Original Research

PUBLISHED 30 January 2023

DOI 10.3389/fvets.2022.1101461

Pharmacokinetic and
OPEN ACCESS pharmacodynamic properties of
pantoprazole in calves
EDITED BY
Nora Mestorino,
National University of La Plata, Argentina

REVIEWED BY
Pierre-Louis Toutain, Jeff D. Olivarez1*, Pierre-Yves Mulon2 , Lisa S. Ebner3 ,
Ecole Nationale Vétérinaire de Toulouse
(ENVT), France Haley Cremerius2 , Channing Cantrell2 , Rebecca Rahn2 ,
Matilde Sierra Vega,
University of Leon, Spain Windy Soto-Gonzalez2 , Joan Bergman4 , Sherry Cox4 ,
*CORRESPONDENCE Jonathan P. Mochel5 , Amanda J. Kreuder6 and Joe S. Smith1,2*
Jeff D. Olivarez 1
[email protected] Biomedical Sciences, College of Veterinary Medicine, Iowa State University, Ames, IA, United States, 2 Large
Joe S. Smith Animal Clinical Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, Knoxville, TN,
[email protected]; United States, 3 College of Veterinary Medicine, Lincoln Memorial University, Harrogate, TN, United States,
4
[email protected] Biomedical and Diagnostic Sciences, University of Tennessee, Knoxville, Knoxville, TN, United States,
5
Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State
SPECIALTY SECTION University, Ames, IA, United States, 6 Veterinary Microbiology and Preventive Medicine, College of Veterinary
This article was submitted to Medicine, Iowa State University, Ames, IA, United States
Veterinary Pharmacology and Toxicology,
a section of the journal
Frontiers in Veterinary Science
Introduction: Development of abomasal ulceration is a large concern, especially
RECEIVED 17 November 2022
ACCEPTED 29 December 2022
within calves; however, there is a paucity of research into the use of gastro protectants
PUBLISHED 30 January 2023 in ruminant species. Proton pump inhibitors, such as pantoprazole, are widely used in
CITATION humans and companion animals. Their efficacy in ruminant species is undetermined.
Olivarez JD, Mulon P-Y, Ebner LS, Cremerius H, The objectives of this study were to 1) estimate the plasma pharmacokinetic
Cantrell C, Rahn R, Soto-Gonzalez W,
parameters for pantoprazole in neonatal calves after three days of intravenous (IV)
Bergman J, Cox S, Mochel JP, Kreuder AJ and
Smith JS (2023) Pharmacokinetic and or subcutaneous (SC) administration, and 2) measure the effect pantoprazole had on
pharmacodynamic properties of pantoprazole abomasal pH over the treatment period.
in calves. Front. Vet. Sci. 9:1101461.
doi: 10.3389/fvets.2022.1101461 Methods: Pantoprazole was administered to 6 Holstein-Angus cross bull calves at a
COPYRIGHT
dose of 1 mg/kg (IV) or 2 mg/kg (SC), once a day (every 24 h) for three days. Plasma
© 2023 Olivarez, Mulon, Ebner, Cremerius, samples were collected over a 72 h period and analyzed via HPLC-UV for determining
Cantrell, Rahn, Soto-Gonzalez, Bergman, Cox, pantoprazole concentrations. Pharmacokinetic parameters were derived via non-
Mochel, Kreuder and Smith. This is an
open-access article distributed under the terms
compartmental analysis. Abomasal (n= 8) samples were collected via abomasal
of the Creative Commons Attribution License cannulas over a 12 h period, per calf per day. Abomasal pH was determined via a bench
(CC BY). The use, distribution or reproduction top pH analyzer.
in other forums is permitted, provided the
original author(s) and the copyright owner(s) Results: Following Day 1 of IV administration, plasma clearance, elimination half-life,
are credited and that the original publication in and volume of distribution of pantoprazole were estimated at 199.9 mL/kg/h, 1.44 h,
this journal is cited, in accordance with
accepted academic practice. No use,
and 0.51 L/kg, respectively. On Day 3 of IV administration, the reported values were
distribution or reproduction is permitted which 192.9 mL/kg/h, 2.52 h, and 1.80 L/kg mL, respectively. Elimination half-life and volume
does not comply with these terms. of distribution (V/F) of pantoprazole following SC administration were estimated at
1.81 h and 0.55 L/kg, respectively, on Day 1; and 2.99 h and 2.82 L/kg, respectively, on
Day 3.
Discussion: The reported values for IV administration were similar to those previously
reported in calves. SC administration appears to be well absorbed and tolerated. The
sulfone metabolite was detectable for 36 h after the last administration for both routes.
Abomasal pH was significantly higher than the pre-pantoprazole pH 4, 6, and 8 h
after administration in both the IV and SC groups. Further studies of pantoprazole as
a treatment/preventative for abomasal ulcers are warranted.

KEYWORDS

bovine, ulcer, abomasum, cattle, proton pump inhibitor

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Olivarez et al. 10.3389/fvets.2022.1101461

1. Introduction at the University of Tennessee. The calves were individually housed

in hutches under a shaded pavilion. Enrollment criteria for this
Ulceration of the gastrointestinal system is a complex disease study included no previous medical history of illness, no previous
that has been documented in many domesticated species (1–5). administration of medication, as well as a physical examination by
Specifically, in cattle, abomasal ulceration is a common cause of a veterinarian that yielded normal vital parameters for a bovine calf.
morbidity and mortality throughout the beef and dairy industries Vaccination status was unknown for all calves. All calves received a
(1, 6). Identified factors contributing to ulceration include stress (e.g., diet of commercial non-medicated milk replacer that either met or
handling, travel), weather, housing, mineral deficiencies, bacterial exceeded the National Research Council (NRC) requirements for the
overgrowth, and the use of non-steroidal anti-inflammatory drugs maintenance and growth of bovine calves. All calves had ad libitum
(NSAIDs) (7). The animal’s age may also contribute to the disorder, access to water.
with the highest reported prevalence in veal calves; however, ulcers Two weeks prior to enrollment in the study, the calves underwent
can be found in all ages of cattle (1, 8). Production of hydrochloric a procedure to place abomasal cannulas (Bard Gastrostomy Buttons;
acid (HCl) by gastric parietal cells has been proven to exacerbate BB) as previously described (20). Briefly, the calves were anesthetized
ulceration and is believed to contribute to the establishment of ulcers using a total intravenous anesthesia (TIVA) protocol of xylazine (0.1
(9). Many pharmaceutical therapies for gastric ulceration in non- mg/kg), butorphanol (0.1 mg/kg), and ketamine (1 mg/kg boluses, as
ruminant species are directed toward the reduction of gastric HCl needed). Immediately prior to surgery, the calves were administered
because of its role in ulcer formation (4, 5, 10). It is proposed that a single dose of flunixin meglumine (1.1 mg/kg, IV) and a single
reducing HCl is therapeutic for ruminants, although the complete dose of procaine penicillin (22,000 IU/kg, IM). Once anesthetized,
etiology of abomasal ulcerations is unknown. the calves were placed in left lateral recumbency. The right paracostal
Pantoprazole is a proton pump inhibitor that irreversibly binds area was clipped and aseptically prepped using povidine surgical
to the hydrogen potassium ATPase pumps on the luminal surface scrub and 70% alcohol. The skin was then infused with 10–15 mLs of
of parietal cells in the gastric mucosa to prevent acid secretion (11). 2% lidocaine in an “inverted-L” pattern, craniodorsal to the incision.
Pantoprazole is labeled for the treatment of peptic ulcers in humans A 5 cm longitudinal incision was then made through the skin and
by reducing acid secretion and increasing gastric pH (12). Increasing underlying muscle layers, starting 3 cm caudal to the rib cage and
gastric pH to >4 for >66% of a 24 h period has been the suggested ∼10 cm lateral to ventral midline. The body of the abomasum was
therapeutic window in promoting the healing of gastric ulcers in then identified and pexied to the body wall using two horizontal
humans and horses (13). Studies in both foals and alpacas have shown mattress sutures at the cranial and caudal portions of the body wall
that intravenous pantoprazole effectively increases gastric pH, but incision. The cannulas were then inserted via a stab incision through
studies in ruminant species are lacking (14, 15). Several retrospective the body wall and into the abomasum and secured with a purse-string
investigations demonstrate the use of pantoprazole in clinical bovine suture. The calves were then recovered, and the surgical sites were
patients, but no pharmacodynamic information exists (16–18). A allowed to heal for 14 days prior to study initiation.
recent study investigating the pharmacokinetic properties of IV
pantoprazole in neonatal calves demonstrated a longer elimination
half-life of pantoprazole in calves and similar clearance compared
to both alpacas and foals, suggesting IV pantoprazole may be an 2.2. Experimental design
effective therapeutic option in ruminants if it possesses similar
pharmacodynamic properties (19). The study of pantoprazole in The six calves were randomly allocated (by coin toss) into
alpacas also indicated subcutaneous administration of pantoprazole either one of two groups: intravenous (IVG) or subcutaneous (SCG)
effectively reduced third compartment (C3) gastric acid secretion administration groups.
(14). There are currently no studies investigating the use of Surgical implantation of all six calves was completed over a 2 day
subcutaneous pantoprazole in ruminants; however, intravenous period, and all animals were given at least 14 days to recover prior
access is not always manageable in cattle, and there is a need for to the start of the experiment. Abomasal contents were collected for a
alternative parenteral routes. 12 h period before pantoprazole administration (day 0), at time points
The primary objectives of this study were to determine 0, 1, 2, 3, 4, 6, 8, and 12 h.
the pharmacokinetic and pharmacodynamic properties of On day 1, 1 h before the initiation of the study, the calves
pantoprazole after multi-day intravenous (IV) and subcutaneous were restrained with a rope halter, and an IV jugular catheter was
(SC) administration of pantoprazole in neonatal calves as well as to placed aseptically in all calves. Preceding catheter placement, the
report the pharmacokinetics of the sulfone metabolite in these calves. skin was aseptically prepared utilizing four alternating wipes of
povidine surgical scrub and 70% isopropyl alcohol. The skin was then
infiltrated with 1–2 mLs of 2% lidocaine, and a #15 blade was used to
2. Materials and methods create a cut-down incision. A 14-gauge catheter was placed through
the cut down into the jugular, and the catheter was secured to the skin
2.1. Animals using 2 Ethilon suture. A second IV catheter was then placed in the
contralateral jugular vein in the IVG calves using the same process
This study was completed at the University of Tennessee’s described above.
Veterinary Research and Education Center. Six Holstein-Angus bull Pantoprazole sodium (West-Ward, Eatontown, NJ,
calves procured from a single source farm were enrolled in the United States) was reconstituted to a 4 mg/mL concentration
study. The age of these calves at enrollment was 17–18 days, with using 10 mL of a 0.9% saline 1L IV bag per manufacturer’s
an average weight of 55.8 ± 4.6 kg. The study was approved by the recommendations. The calves were fed ∼2 h prior to T0 and at 2 h
Institution Animal Care and Use Committee (IACUC # 2825-05221) prior to T12. At T0, the IVG calves received a 1 mg/kg dose in the

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left jugular vein over 1 min, while the SCG calves were administered ∼30 mm in order to bypass the one-way valve within the tube.
a 2 mg/kg dose subcutaneously in the neck. SC doses >10 mL were Negative pressure was then gently applied using the 12 mL syringe
administered in more than one location, per Beef Quality Assurance until 4–5 mL of abomasal contents were acquired. The pH of each
(BQA) guidelines. Pantoprazole administration for the IVG and SCG sample was then recorded within 15 min of collection in a process
calves was repeated at 24 h intervals on days 2 and 3. described below.
On days 1 and 3 of the study, blood samples were collected at The aliquots of abomasal content were placed into a 30 mL
0, 15, 30, 45, and 60 min, as well as 2, 3, 4, 8, 12, 18, and 24 h after falcon tube. A benchtop pH analyzer (UB-10 pH/mV meter, Denver
drug administration. Samples were obtained from all calves through Instruments, US) was then used to measure pH. The analyzer was
an IV catheter in the right jugular vein utilizing a previously described calibrated prior to each sample set according to the manufacturer’s
push-pull technique (21). The blood samples were immediately procedure. Once calibrated, the probe was introduced into the
placed into sodium heparin tubes, then placed on ice until centrifuged abomasal content sample and allowed to equilibrate for 30 s, at which
at 1,500 × g for 10 min. All blood samples were processed within 6 h time the pH was recorded.
of collection. After centrifugation plasma was transferred to cryovials
which were then stored at −80◦ C until analysis. Abomasal contents
were collected each day of the study at 0, 1, 2, 3, 4, 6, 8, and 12 h after 2.4. Analytical method
pantoprazole administration and processed as described below.
After a 10 day washout period, calves originally in the IVG were Plasma pantoprazole analysis was performed using a reverse
switched to the SCG and vice versa. The study was then repeated as phase high performance liquid chromatography (HPLC) method as
described above. previously described for pantoprazole and the pantoprazole sulfone
metabolite in goat plasma (22). The system consisted of a 2,695
separations module and a 2,487 UV absorbance detector (Waters).
The compounds were separated on a Symmetry C18 (4.6 × 150 mm,
2.3. Abomasal content collection and pH 5 µm) column with a 5 µm Symmetry C18 guard column. The
measurement mobile phase was a mixture of 0.1 M sodium phosphate dibasic and
acetonitrile (68:32). The flow rate was 1 mL/min, and absorbance was
Abomasal contents were collected via the surgically placed BB measured at 290 nm.
tube. Briefly, a 3 × 70 mm stainless steel two-eyed teat cannula Pantoprazole and its metabolite were extracted from plasma
attached to a 12 mL syringe was introduced into the BB tube samples using a liquid-liquid extraction method. Previously frozen

TABLE 1 Day 1 pharmacokinetic parameters of pantoprazole after intravenous (IV) and subcutaneous (SC) administration in calves.

Compound (route) Parameter Unit Mean Standard deviation Median Min Max
Pantoprazole (IV) C0 ng/mL 2,147 257 2,190 1,815 2,519
∗
AUClast ng/mL h 3,422 1,020 4,008 1,878 4,351
∗
AUCinf ng/mL h 3,586 1,100 4,089 1,945 4,632
∗
AUMCinf ng/mL h 5,372 4,568 6,774 1,207 13,881

MRTinf h 1.95 1.2 2.08 0.76 4.1

Cl mL/h/kg 199.9 89.7 175.3 154.8 368.6

λz 1/h 0.34 0.37 0.31 0.13 1.02

T1/2 (λz) h 1.44 1.29 2.25 0.68 3.9

Vz L/kg 0.51 0.27 0.57 0.24 0.88

Pantoprazole (SC) Cmax ng/mL 3,435 771 3,436 2,350 4,553

Tmax h 0.58 0.21 0.75 0.25 0.75

∗
AUClast ng/mL h 7,629 3,832 7,070 4,270 14,646
∗
AUCinf ng/mL h 7,857 3,849 7,402 4,502 14,894
∗
AUMCinf ng/mL h 21,046 20,818 18,800 12,014 66,608

MRTinf h 2.67 1.45 2.22 1.7 5.11

λz 1/h 0.34 0.21 0.38 0.17 0.75

T1/2 (λz) h 1.81 1.37 1.86 0.92 4.13

Vz /F L/kg 0.55 0.61 0.60 0.22 1.90

C0 , calculated concentration at time zero of IV administration; AUClast , area under the curve calculated at the last time point; AUCinf , area under the curve extrapolated to infinity; AUMCinf , area
under the moments curve extrapolated to infinity; MRTinf , mean residence time extrapolated to infinity; Cl, plasma clearance; T1/2 (λz), elimination half-life [reported as harmonic mean (34, 35)];
Vz , volume of distribution; Cmax , maximum plasma concentration after SC administration; Tmax , time to reach maximum plasma concentration after SC administration; Vz /F, volume of distribution
accounting for bioavailability; λz, elimination rate constant. Means reported as Geometric mean, with the exception of elimination half-life, which is reported as harmonic mean.

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samples were thawed, vortex-mixed, and 100 µl of plasma was time vs. concentration information for pantoprazole was determined
transferred to a 13 × 100 mm screw top tube, followed by 10 µl via HPLC from the samples collected at 8 time points ranging
of tinidazole (internal standard, 100 µg/mL) and 2 mL chloroform. from 0 to 12 h after Day 1 and 3 of administration. Standard
The tubes were rocked for 15 min and then centrifuged for PK parameters were generated for individual calves, as follows:
20 min at 1,000 × g. The organic layer was transferred to a Maximum concentration of pantoprazole extrapolated to time zero,
glass tube and evaporated to dryness with nitrogen gas. Samples C0; Time of maximum pantoprazole concentration, Tmax; Area
were reconstituted in 250 µL of mobile phase, and 100 µL under pantoprazole concentration–time curve, AUClast and AUCinf;
was analyzed. Area under the moment curve, AUMCinf; Pantoprazole mean
Standard curves for the plasma analysis were prepared residence time, MRT = AUMCinf/AUCinf; Pantoprazole terminal
by fortifying untreated, pooled plasma with pantoprazole and half-life, T1/2 (λz) = ln (2)/λz; Pantoprazole systemic clearance,
the metabolite, which produced a linear concentration range CL = Dose/AUCinf; Volume of distribution of pantoprazole
of 0.01–100 µg/mL. The average recovery for pantoprazole (area), Vz.
and it’s metabolite was 100 and 90%. The average recovery A linear/log trapezoidal rule was used for data analysis to estimate
for the internal standard was 99%. The quality control the area under the pantoprazole time-curve. Summary statistics on
(QC) samples used for validation were 0.03, 0.3, 3, and the individual PK parameters were performed thereafter to derive the
30 µg/mL, and the intra and inter-assay variability ranged geometric mean, median, minimum, and maximum.
from 2 to 11% for pantoprazole and 3% to 9% for the Bioavailability (F) was calculated utilizing the following equation:
metabolite. The lower limit of quantification for both was
0.1 µg/mL. AUC(SC) Dose(IV)
F= x
AUC(IV) Dose(SC)

2.5. Pharmacokinetic analysis

2.6. Statistical analysis
Pharmacokinetic parameters were calculated from time-plasma
concentration data as previously described (19). Pharmacokinetic Data from pH testing was evaluated for normality. Then
modeling was performed via standard industry modeling software one-way ANOVA followed by multiple comparisons (based on
(PKanalix, Monolix Suite 2021R2, Lixoft, France) as described for distribution) testing was performed using GraphPad Prism (version
pantoprazole in goats and calves (19, 23). Standard data representing 8.0.0, GraphPad Software, San Diego, CA). Day zero was the baseline

TABLE 2 Day 3 pharmacokinetic parameters of pantoprazole after intravenous (IV) and subcutaneous (SC) administration in calves.

Compound (route) Parameter Unit Mean Standard deviation Median Min Max
Pantoprazole (IV) C0 ng/mL 2,575 371 2,592 2,105 3,214
∗
AUClast ng/mL h 4,580 1,123 4,635 2,933 5,899
∗
AUCinf ng/mL h 5,171 994 5,257 3,500 6,212
∗
AUMCinf ng/mL h 27,338 21,737 24,098 12,058 68,709

MRTinf h 5.29 4.11 5.22 2.36 12.91

Cl mL/h/kg 192.9 36.5 188.1 158.1 255.0

λz 1/h 0.11 0.13 0.078 0.047 0.35

T1/2 (λz) h 2.52 1.86 2.71 1.99 6.91

Vz L/kg 1.80 1.96 2.65 0.54 4.67

Pantoprazole (SC) Cmax ng/mL 4,221 604 4,326 3,439 5,085

Tmax h 0.41 0.21 0.5 0.25 0.75

∗
AUClast ng/mL h 8,797 2,617 9,289 5,010 11,512
∗
AUCinf ng/mL h 9,417 2,789 9,838 5,585 12,336
∗
AUMCinf ng/mL h 45,031 26,684 47,165 20,159 84,666

MRTinf h 4.78 1.75 5.79 2.26 6.86

λz 1/h 0.077 0.094 0.057 0.053 0.27

T1/2 (λz) h 2.99 0.87 3.63 1.76 3.79

Vz /F L/kg 2.82 2.06 3.08 0.79 6.52

C0 , calculated concentration at time zero of IV administration; AUClast , area under the curve calculated at the last time point; AUCinf , area under the curve extrapolated to infinity; AUMCinf , area
under the moments curve extrapolated to infinity; MRTinf , mean residence time extrapolated to infinity; Cl, plasma clearance; T1/2 (λz), elimination half-life; Vz , volume of distribution, Cmax ,
maximum plasma concentration after SC administration; Tmax , time to reach maximum plasma concentration after SC administration; Vz /F, volume of distribution accounting for bioavailability;
λz, elimination rate constant. Means reported as Geometric mean, with the exception of elimination half-life, which is reported as harmonic mean.

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FIGURE 1
Time vs. Concentration curves for intravenous (IV; square) and subcutaneous (SC; triangle) administration of pantoprazole on Days 1 (top) and 3 (bottom).

being compared to individual treatment days. Significance was set at cannulas remaining in place throughout the entire study period
a value of P < 0.05. and only mild swelling noted at the insertion site. One BB came
dislodged after day 1 of the first trial and another become dislodged
immediately prior to the beginning of the first trial. These calves
3. Results were kept in the study for pharmacokinetic data and was included
in the days 0 and 1 abomasal pH data. One of the calves
3.1. Animals developed a catheter site infection and thrombophlebitis at the IV
catheter sites. Intravenous catheterization was well-tolerated by all
At the time of enrollment, all calves had normal vital parameters other calves. No clinical manifestations of adverse reactions were
and were considered healthy. Surgical implantation of the BB observed due to either intravenous or subcutaneous administration
was well-tolerated by most of the calves, with four of the six of pantoprazole.

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TABLE 3 Day 1 pharmacokinetic parameters of pantoprazole sulfone after intravenous (IV) and subcutaneous (SC) administration of pantoprazole sodium in
calves.

Compound (route) Parameter Unit Mean Standard deviation Median Min Max
Pantoprazole sulfone (IV) Cmax ng/mL 105 41 103 60 162

Tmax h 2.45 1.03 3 1 4

∗
AUClast ng/mL h 1,131 707 1,355 407 2,415

AUCinf ng/mL∗ h 1,472 861 1,749 647 2,867

AUMCinf ng/mL∗ h 18,954 15,328 26,653 67,867 39,042

MRTinf h 12.85 3.57 12.89 8.46 18.49

λz 1/h 0.084 0.024 0.085 0.058 0.12

T1/2 (λz) h 8.0 2.21 8.17 5.92 11.9

Pantoprazole sulfone (SC) Cmax ng/mL 231 91 213 138 351

Tmax h 3.57 0.55 4 3 4

∗
AUClast ng/mL h 2,908 1,857 3,188 1,146 5,546
∗
AUCinf ng/mL h 3,657 3,006 4,576 1,273 8,767
∗
AUMCinf ng/mL h 50,251 77,571 84,165 10,330 203,632

MRTinf h 13.74 6.84 14.2 8.12 23.23

λz 1/h 0.085 0.053 0.08 0.048 0.17

T1/2 (λz) h 7.2 4.62 8.7 4.12 14.4

AUCinf , area under the curve extrapolated to infinity; AUMCinf , area under the moments curve extrapolated to infinity; MRTinf , mean residence time extrapolated to infinity; T1/2 (λz), elimination
half-life; Cmax , maximum plasma concentration; Tmax , time to reach maximum plasma concentration; λz, elimination rate constant. Means reported as Geometric mean, with the exception of
elimination half-life, which is reported as harmonic mean.

TABLE 4 Day 3 pharmacokinetic parameters of pantoprazole sulfone after intravenous (IV) and subcutaneous (SC) administration of pantoprazole sodium in
calves.

Compound (route) Parameter Unit Mean Standard deviation Median Min Max
Pantoprazole sulfone (IV) Cmax ng/mL 153 116 147 63 401

Tmax h 2.14 1.03 2 1 4

∗
AUClast ng/mL h 2,273 1,340 2,611 678 4,527
∗
AUCinf ng/mL h 2,743 1,661 3,434 780 4,968
∗
AUMCinf ng/mL h 47,460 47,852 60,600 7,438 123,356

MRTinf h 17.34 7.66 16.68 9.53 29.03

λz 1/h 0.061 0.03 0.064 0.035 0.12

T1/2 (λz) h 10.5 5.39 10.98 5.92 19.66

Pantoprazole sulfone (SC) Cmax ng/mL 383 181 326 272 712

Tmax h 2.77 0.45 3 2 3

∗
AUClast ng/mL h 5,834 3,048 5,312 3,288 10,870
∗
AUCinf ng/mL h 7,086 4,321 6,210 3,708 13,924
∗
AUMCinf ng/mL h 141,436 125,690 117,261 58,146 325,838

MRTinf h 19.96 4.77 18.88 15.68 27.09

λz 1/h 0.051 0.013 0.055 0.039 0.067

T1/2 (λz) h 12.8 3.49 12.7 10.4 17.94

AUCinf , area under the curve extrapolated to infinity; AUMCinf , area under the moments curve extrapolated to infinity; MRTinf , mean residence time extrapolated to infinity; T1/2 (λz), elimination
half-life; Cmax , maximum plasma concentration; Tmax , time to reach maximum plasma concentration; λz, elimination rate constant. Means reported as Geometric mean, with the exception of
elimination half-life, which is reported as harmonic mean.

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FIGURE 2
Time vs. Concentration curves for pantoprazole sulfone after intravenous (IV; square) and subcutaneous (SC; triangle) administration of pantoprazole on
Days 1 (top) and 3 (bottom).

3.2. Pharmacokinetics pharmacokinetic parameters of pantoprazole in calves after IV

and SC administration. IV and SC administration had fairly rapid
No concentrations of pantoprazole were detected in any of elimination with half-lives of 1.44 and 1.81 h, respectively. Table 2
the calves prior to administration on Day 1. Table 1 displays displays the geometric mean, median, minimum, and maximum of
the geometric mean, median, minimum, and maximum of the the pharmacokinetic parameters of pantoprazole in calves after three

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FIGURE 3
Comparison of abomasal pH over time between control, IV, and SC treatment groups on Day 1. Black circles, pH pre-pantoprazole administration;
Orange triangles, pH after IV pantoprazole administration; Gray square, pH after SC administration. Statistical significance (P < 0.05) noted with asterisk.

consecutive days of IV and SC administration. The elimination half- intravenous and subcutaneous administration of pantoprazole in
lives of intravenous and subcutaneous administration had increased calves. In this study, both IV and SC administration of pantoprazole
to 2.52 and 2.99 h, respectively. Figure 1 displays the time vs. maintained abomasal pH to > 4 in calves over a 12 h period.
concentration curves for pantoprazole on day 1 and 3. The pharmacokinetic properties of pantoprazole after 1 day of
Tables 3, 4 display the geometric mean, median, minimum, IV administration were less than those previously reported (19). The
and maximum of the pharmacokinetic parameters of the sulfone half-life of pantoprazole reported in that study was 2.81 h, while
metabolite on days 1 and 3, respectively. Figure 2 displays the time our current study found a half-life of 1.44 h. Differences between
vs. concentration curves for the sulfone metabolite on days 1 and 3. these values could be due to variations between measurement
Bioavailability of the first SC administration was 115.2 ± 56.0 %. techniques as well as differences in lower limits of quantifications
(LLOQ). The LLOQ for the original pharmacokinetic study was
reported to be much lower than our current study’s (0.002 and
3.3. pH investigation 0.1µg/mL, respectively). The ability to detect smaller amounts of
pantoprazole in the plasma could account for the longer half-life (24).
Figures 3–5 compare the abomasal pH between the control and For subcutaneous administration, the Cmax (3,434.77 ng/mL) was
treatment groups on days 1–3, respectively. Of note, both treatment achieved fairly rapidly (within 0.58 h), indicating the pantoprazole
groups had a significantly higher pH at the 4, 6, and 8 h time was rapidly absorbed, which is also reported in human studies (25).
points compared to the control pretreatment pH on all 3 days. Interestingly the pharmacokinetic data for day 3 demonstrated
Tables 5, 6 display the average pH at each time point for IV and SC potential accumulation of the drug with multiple-day administration,
pantoprazole, respectively. as Cmax, AUC, MRT, and T1/2 had all increased for both IV
and SC groups. However, due to the short elimination half-lives,
this could be non-linear kinetics in disposition, possibly due to
4. Discussion enzymatic inhibition. Pantoprazole is rapidly metabolized by the
liver’s cytochrome P450 (CYP) system (26). Previous studies have
To our knowledge, this is the first report investigating the indicated that some PPIs [such as omeprazole (27)] can inhibit
pharmacokinetic and pharmacodynamic properties of multi-day the CYP, reducing its efficacy in breaking down the drug into its

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FIGURE 4
Comparison of abomasal pH over time between control, IV, and SC treatment groups on Day 2. Black circles, pH pre-pantoprazole administration;
Orange triangles, pH after IV pantoprazole administration; Gray square, pH after SC administration. Statistical significance (P < 0.05) noted with asterisk.

metabolites, meaning the parent drug can be detected in higher though more time points would be needed to confirm this timeframe.
concentrations after repeat administrations; however, pantoprazole While these results are promising in calves, the efficacy of their use
has not been shown to inhibit the CYP in humans (28). If this is in adult ruminants is unclear due to vastly different gastrointestinal
true in ruminants, then increases in the pharmacokinetic values on physiology. As calves are considered pre-ruminants, they function
Day 3 may be due to the saturation of the CYP mechanism rather similarly to monogastric species because the esophageal groove allows
than inhibition. Despite increased pantoprazole concentrations in the milk to bypass the rudimentary rumen and enter directly into the
plasma, there was no significant difference in abomasal pH between abomasum (29, 30). Adult ruminants have a functional rumen, which
Day 1 and Day 3 of treatment. is a constant source of ingesta that enters the abomasum. As food
On all 3 days of testing, the abomasal pH was significantly higher entering the abomasum is a stimulant for acid secretion, the near-
than the pre-pantoprazole pH 4, 6, and 8 h after administration in constant movement of ingesta from the reticulo-rumen into the
both the IVG and SCG. As seen in Figures 3–5, the pretreatment abomasum suggests a more prolonged period of acid production
pH steadily declines during the first 4 h of the study, then maintains when compared to calves (31). Prolonged acid secretion may present
a pH of around 2.0 from the 4–8 h period. The calves were fed a a challenge for PPI therapy, and modifications may need to be made
commercial milk replacer ∼2 h before the 0 h and the 12 h time in dosing regimens.
points. The pH of the milk replacer was ∼6.61 ± 0.046, which may The sulfone metabolite was detected in all calves after all
account for the relatively high abomasal content pH at 0 h and the administrations, with concentrations detectable at 24 and 36 h after
drastic increase in pH of the control calves between the 8 and 12 h day 1 and 3 dosing, respectively. This is similar to the tissue
time points. In human and equine medicine, achieving a gastric concentrations of pantoprazole sulfone detected in calves at 1–3
pH of >4 for >66% of a 24 h period is ideal for the healing of days after intravenous administration (19), but different from goats
gastric ulcers (13). Our study’s data suggest that both IV and SC which had no detectable levels of pantoprazole sulfone after 4 h
administration of pantoprazole can achieve this therapeutic window, post intravenous administration at 1 mg/kg (23). While the sulfone

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FIGURE 5
Comparison of abomasal pH over time between control, IV, and SC treatment groups on Day 3. Black circles, pH pre-pantoprazole administration;
Orange triangles, pH after IV pantoprazole administration; Gray square, pH after SC administration. Statistical significance (P < 0.05) noted with asterisk.

TABLE 5 Comparison of average hourly abomasal pH after intravenous TABLE 6 Comparison of average hourly abomasal pH after subcutaneous
administration of pantoprazole. administration of pantoprazole.

Time (h) Day 0 Day 1 Day 2 Day 3 Time (h) Day 0 Day 1 Day 2 Day 3
0 5.778 5.912 6.025 5.785 0 5.765 5.825 5.95 5.935

1 5.366 5.754 5.6875 5.79 1 5.5175 5.7725 5.9425 5.795

2 4.546 5.872 5.4825 5.5625 2 5.0025 5.79 5.8625 5.9525

3 3.972 6.264 5.635 5.65 3 4.5275 5.905 5.95 5.8875

c a b a a
4 2.452 6.54 5.7825 5.5875 4 2.79 6.005 5.8075 5.82a

6 1.86 6.35c 5.76b 5.945b 6 1.73 5.97c 5.605c 5.6375c

8 2.018 6.196a 5.9025a 6.0225a 8 2.1 5.5425c 6.17c 5.65b

12 5.334 6.286 5.1675 5.295 12 4.8725 5.4125 5.3425 5.3925

a Indicates statistically significant difference (p-value of 0.05–0.01) from the control (Day 0). a Indicates statistically significant difference (p-value of 0.05–0.01) from the control (Day 0).
b Indicates statistically significant difference (p-value of 0.01–0.005) from the control (Day 0). b Indicates statistically significant difference (p-value of 0.01–0.005) from the control (Day 0).
c Indicates statistically significant difference (p-value of <0.005) from the control (Day 0). c Indicates statistically significant difference (p-value of <0.005) from the control (Day 0).

metabolite is thought to be inactive, it is detectable for longer in 0.54 ± 0.01 1/h after subcutaneous administration on day 3. Figure 6
tissues than the parent compound in calves (19), so future studies displays the simultaneous time vs. concentration data for the parent
investigating tissue residue disposition could further determine the and metabolite for each day and method of administration. From
relationship between plasma and tissue pantoprazole sulfone levels. these values it appears that the formation of the sulfone metabolite is
Arithmetic mean ± SD values of elimination rate constants (λz) the rate limiting step in the pharmacokinetics of pantoprazole sulfone
for parent and sulfone metabolite were 0.47 ± 0.36 and 0.087 ± 0.024 in calf. When evaluating the arithmetic mean ± SD λz for IV and SC
1/h after intravenous administration on day 1; 0.38 and 0.08 1/h after parent on day one values are overall comparable at 0.47 ± 0.36 and
subcutaneous administration on day 1; 0.15 ± 0.13 and 0.066 ± 0.03 0.36 ± 0.22 1/h, respectively, while on day 3 these values are 0.15 ±
1/h after intravenous administration on day 3; and 0.099 ± 0.094 and 0.13 and 0.099 ± 0.094 1/h. Comparisons of λz for the metabolite

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FIGURE 6
Comparison of time vs. concentration for parent and metabolite for each day and route of administration.

after IV and SC administration are 0.087 ± 0.024 and 0.097 ± 0.053 as genetic polymorphisms. The animals in this study were also all
1/h on day 1, and 0.066 ± 0.03 and 0.054 ± 0.01 1/h on day 3. from a single farm and all healthy, which is not reflective of the
While there are subtle differences between the slopes of pantoprazole population of animals typically presenting to a hospital for care.
after IV and SC administration, the similarities in elimination do not Future research should investigate the efficacy and safety of
support flip flop pharmacokinetics for SC administration. pantoprazole in sick populations, as well as investigate its use in
After the first SC administration, bioavailability (F) was noted to adult ruminants. As this study has demonstrated that the abomasal
be 115%. While uncommon, a bioavailability of >100% can occur pH can be increased with pantoprazole, it would be imperative
for several reasons, such as distribution between compartments, to determine the effect on the microbiome of the gastrointestinal
or as a result of sampling schedule. A bioavailability of >100% tract. Further investigations into the ideal dosage and frequency
has been observed for pantoprazole in alpacas (115%) (14), as would also be important as therapy with pantoprazole can represent
well as a similar proton pump inhibitor esomeprazole in goats a substantial economic investment. As ruminants are part of the
(F = 116%) (32). This could indicate non-linear pharmacokinetics food supply and the use of pantoprazole is considered extra-label,
at higher dosages or potentially flip flop pharmacokinetics, although establishing appropriate withdrawal times is vital. Additionally, while
flip flop pharmacokinetics seems less likely comparing the slopes of pH timepoints were collected at 18 hours, these were not presented,
each route. as it appears that additional sampling of pH after the second milk
All the calves tolerated both routes of administration well, and feeding should be considered for future studies in calves to truly
no pain or swelling was noted at the injection sites. Adverse events account for pH changes from pantoprazole, vs. the second milk
in humans include diarrhea, headache, and abdominal pain (33). feeding. This model of cannulation for abomasal fluid sampling could
All calves in the study maintained a healthy appetite and fecal be utilized for other gastroprotectant therapies in ruminants, such as
consistency. Previous safety investigations of pantoprazole use in esomeprazole, the S-enantiomer of omeprazole (32). Future studies
hospitalized ruminants also report similar findings (16). could also evaluate the variation within individuals of a population to
The major limitation of this study is the small sample size that see if factors such as disease status or genetic polymorphism influence
was used. While pharmacokinetic studies are appropriately powered the metabolism of pantoprazole.
with 4–6 animals, increasing the number of animals can decrease the In conclusion, pantoprazole effectively increased abomasal pH
impact of outlier data, as well as to highlight potential population in calves after either IV or SC administration, and multiple-day
variables which may describe the variation in some parameters, such administration appeared to be well-tolerated. While further research

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is needed to determine the role of acid secretion in abomasal ulcer contributed to manuscript development. All authors contributed to
formation, this information is a step forward in ulcer management the article and approved the submitted version.
of ruminants.

Acknowledgments
Data availability statement
The authors wish to thank Lainey Harvill, Sarah Bullock, and
The original contributions presented in the study are included in Carla Hill for their assistance in data collection and animal care.
the article/supplementary material, further inquiries can be directed
to the corresponding authors.
Conflict of interest
Ethics statement The authors declare that the research was conducted in the
absence of any commercial or financial relationships that could be
The animal study was reviewed and approved by Institutional construed as a potential conflict of interest.
Animal Care and Use Committee, University of Tennessee.

Publisher’s note
Author contributions
All claims expressed in this article are solely those of the
JO, P-YM, LE, AK, JM, and JS devised the experimental design. authors and do not necessarily represent those of their affiliated
SC and JB developed the analytical method. JO, P-YM, LE, JS, HC, organizations, or those of the publisher, the editors and the reviewers.
CC, RR, and WS-G contributed to animal setup, husbandry, and data Any product that may be evaluated in this article, or claim that may
collection. JO, SC, JB, and HC contributed to sample analysis. JO, be made by its manufacturer, is not guaranteed or endorsed by the
AK, JM, SC, and JS contributed to data interpretation. All authors publisher.

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