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Zuprevo® – A new paradigm in the treatment and prevention of BRD
6 th European Congress of
Bovine Health Management
7 – 9 September 2011
Liège, Belgium
SummitAngers
Schwabenheim & Bayerseich
Pharmaceutical R & D at MSD Animal Health
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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Contents1
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Preface ���������������������������������������������������������������������������������������������������������������������������4
Curriculum�Vitae������������������������������������������������������������������������������������������������������������ �6
Tildipirosin�–��
Mode�of�Action�of�a�Novel�Macrolide� ����������������������������������������������������������������������������9
Zuprevo®:�
Key�features�of�a�unique�product� �������������������������������������������������������������������������������� �15
Preventive�efficacy�of�Zuprevo®�in�a�Mannheimia haemolytica�
challenge�study�of�bovine�respiratory�disease� �������������������������������������������������������������� �23
Zuprevo®:�Efficacy�in�the�treatment�of�BRD�in�calves�
in�comparison�to�Draxxin®� ������������������������������������������������������������������������������������������ �31
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
The new MSD Animal Health is delighted to welcome you to our symposium at the
6th European Congress of Bovine Health Management in this beautiful setting along
the river Meuse in the heart of Liège. On this occasion, we are pleased to introduce a
new tool called “Zuprevo®” which will help veterinarians and cattle producers around
the world to be in control of bovine respiratory disease (BRD).
Scientists of MSD Animal Health (formerly Intervet / Schering-Plough Animal Health) in
cooperation with their partners in academia and contract research organizations have
played a crucial role in the discovery and development of this novel tool.
Although bovine veterinarians have many choices available for treatment and prevention,
BRD remains a very frequent and economically important disease in cattle. BRD is a
multi-factorial disease, and cattle are prone to rapidly develop severe lung pathology.
Factors triggering BRD outbreaks include the anatomical predisposition of the bovine
respiratory system, common stress inducing cattle management practices such as weaning,
commingling, and transportation, as well as suboptimal housing and climatic conditions.
Furthermore, the bacterial respiratory pathogens are ubiquitous and appear unlikely
to ever be eradicated.
Findings from advanced diagnostic procedures, specifically the monitoring of intra-ruminal
temperature, suggest that visual detection of BRD very often occurs when the disease
is already well advanced. The initiation of curative treatment is thus often late while
at the same time BRD pathogens may have been rapidly spreading amongst the
untreated animals in the herd. Therefore, for both treatment and prevention, it is
important for the antimicrobial to be present at the site of infection, i. e. lung tissue and
bronchial fluid, both rapidly and for an extended period of time.
Zuprevo® is a novel tri-basic 16-membered-ring macrolide with unique pharmacokinetic
and pharmacodynamic characteristics, available in a highly convenient cattle specific
formulation, addressing both the need for speed in the treatment and for duration in
the prevention of BRD.
Preface
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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In collaboration with MSD Animal Health, Professor Stephen Douthwaite, a molecular
microbiologist from the University of Southern Denmark, Odense, intensively studied the
mode of action of the active ingredient of Zuprevo®, tildipirosin. In his presentation,
he will reveal well illustrated findings based on footprinting experiments and molecular
modeling as a basis of the improved antibacterial activity of tildipirosin and its unique
pharmacodynamic properties.
The key features of the Zuprevo® product profile will be presented by Dr. Rainer Röpke.
He managed the research and development activities for Zuprevo® as global project
manager at MSD Animal Health. Besides highlighting the benefits of a convenient
formulation, a high margin of safety and a comparatively short withdrawal period,
Dr. Röpke will discuss the unique pharmacokinetic profile of tildipirosin in lung tissue
and bronchial fluid, which contributes to the excellent clinical efficacy demonstrated
in field trials across the EU.
Dr. Markus Rose, Senior Development Manager at MSD Animal Health, will describe
the findings from a Mannheimia�haemolytica challenge study demonstrating the
principle of Zuprevo®’s preventive efficacy in BRD, in comparison to Draxxin®. The
superior clinical and antibacterial efficacy of Zuprevo® observed in this study confirms
its outstanding pharmacodynamic and pharmacokinetic properties.
Professor Kerstin E. Müller from the Faculty of Veterinary Medicine at the Free University
of Berlin will present a field trial demonstrating the excellent therapeutic efficacy
of Zuprevo® against BRD in young calves, in comparison to Draxxin®. This study confirms
the fast onset of action and provides further compelling clinical evidence for the
excellent clinical efficacy of Zuprevo® in a real world setting.
MSD Animal Health is committed to be a leader in innovative research and to be a
partner to veterinary practitioners and livestock producers globally. We are therefore
very pleased to be your host at this symposium and to share with you this information
on Zuprevo®, a new standard in the treatment and prevention of BRD.
We wish you a very enjoyable evening.
Dr. Peter Schmid
Head of Drug Discovery
MSD Animal Health
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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Prof. Stephen Douthwaite
Dr. Rainer Röpke
Professor�Stephen�Douthwaite�studied�biochemistry�and�microbiology�at�the�University�
of�Wales�in�Cardiff�(1976)��Soon�afterwards,�he�joined�the�Max�Planck�Institute�in�
West�Berlin�where�he�worked�on�ribosomal�protein�synthesis�and�his�research�interests�
developed�to�include�antibiotic�inhibitors�of�this�process��He�obtained�his�PhD�from�
Aarhus�University�in�Denmark�(1982),�and�spent�three�years�as�a�postdoc�at�the�University�
of�California�in�Santa�Cruz��He�is�presently�Professor�of�Molecular�Microbiology�at�the�
University�of�Southern�Denmark,�Odense,�where�he�has�been�employed�since�1987��
Dr��Rainer�Röpke�studied�Agriculture�at�the�Technical�University�of�Munich�and�the�
University�of�Göttingen,�Germany,�where�he�earned�his�diploma�in�1989��In�1992��
he�obtained�his�PhD�from�the�Institute�for�Animal�Physiology�and�Research�Centre�
for�Milk�and�Food�at�the�Technical�University�of�Munich��Starting�in�the�animal�
health�industry�in�1992�he�worked�as�Global�Regulatory�Affairs�Manager�responsible�
for�Feed�Additives�at�Hoechst�Roussel�Vet�in�Wiesbaden,�Germany,�before�relocating��
in�2000�to�join�their�US�organization�(then�Intervet�Inc�)�in�Delaware,�USA,�as�Director�
Pharmaceutical�Development��Since�2003�he�has�been�Global�Project�Manager�at�
Intervet�Innovation�GmbH�in�Schwabenheim,�Germany�(now�member�of�the�MSD�
Animal�Health�Group),�managing�all�activities�in�relation�to�the�global�development��
of�new�pharmaceutical�products�
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
Dr. Markus Rose
Prof. Dr. Kerstin E. Müller
Dr��Markus�Rose�graduated�from�the�veterinary�faculty�of�the�Justus-Liebig-University�
(JLU),�Gießen,�Germany,�in�1987,�and�worked�in�private�veterinary�practice�till�1989��
Between�1990�and�1993�he�was�research�associate�at�the�Veterinary�Institute�of�the�
Karl-August-University,�Göttingen,�Germany,�specializing�in�microbiology��He�received��
his�doctorate�in�1991��During�his�employment�with�Hoechst�Roussel�Vet�GmbH,�Wiesbaden,�
Germany,�beginning�in�1994,�he�managed�clinical�study�programs,�and�relocated��
to�the�US�in�1998,�finally�serving�as�senior�project�leader��Back�in�Germany�in�2002,�
at�Intervet�Innovation�GmbH�(now�member�of�the�MSD�Animal�Health�Group),��
he�continued�with�project�development,�and�was�responsible�for�a�group�providing�
clinical�study�support��Since�2004�he�has�been�senior�development�manager�with�
focus�on�microbiological,�food�safety�and�clinical�aspects�of�global�pharma�projects�
Professor�Kerstin�E��Müller�earned�her�degree�from�the�Veterinary�University�Hannover�in�
1984��She�obtained�her�doctoral�degree�in�1986�and�was�recognized�as�German�cattle�
specialist�in�1989�when�she�was�member�of�the�Cattle�Clinic�in�Hannover��In�1995�she�
obtained�the�PhD�from�Utrecht�University,�NL,�where�she�was�Senior�Lecturer�for�
Internal�Medicine�of�Ruminants�and�Horses�from�1991�to�1998�and�subsequently�
member�of�the�Department�of�Farm�Animal�Health�with�certification�as�a�Dutch�Cattle�
Specialist��Since�2003�she�has�been�Professor�and�Managing�Director�at�the�Clinic�for�
Ruminants�and�Swine�at�the�Faculty�of�Veterinary�Medicine,�Freie�Universität�Berlin��
She�is�chair�of�the�Education�and�Residency�Committee�of�the�ECBHM�and�of�the�German�
Buiatrics�Association�and�board�member�of�the�WBA�
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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1.Tildipirosin – Mode of Action of a Novel Macrolide
Stephen Douthwaite, Jacob Poehlsgaard and Ralf Warrass
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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1
Tildipirosin – Mode of Action of a Novel MacrolideStephen�Douthwaite1,�Jacob�Poehlsgaard1�and�Ralf�Warrass2
1 Department of Biochemistry & Molecular Biology, University of Southern Denmark, Campusvej 55, DK-5230 Odense M, Denmark.
2 Drug Discovery, Intervet Innovation GmbH, a member of the MSD Animal Health Group, Zur Propstei, D-55270 Schwabenheim, Germany.
Abstract
MSD Animal Health has recently been granted market-
ing authorization in the European Union for Zuprevo®
in the treatment and prevention of respiratory disease
in cattle. The active ingredient tildipirosin is the first
tri-basic 16-membered-ring macrolide on the market.
The lipophilic – hydrophilic flexibility of the three basic
centres of til dipirosin are made responsible for its high
solubility, rapid tissue distribution, extensive accumu-
lation in lung tissue and excellent antibacterial activity
against bacteria associated with bovine respiratory
disease. Tildipirosin’s antimicrobial activity is achieved
by effective penetration of the bacterial cell wall
and binding to the molecular target, the bacterial
ribosome. Using chemical footprinting experiments
and molecular modelling we compared tildipirosin’s
ribosomal attachment to that of other macrolides
(tylosin, tilmicosin, tulathromycin) and found common
inter action sites but also clear differences and unique
binding modes. In an in vitro transcription / translation
assay, tildipirosin demonstrated highly efficient
inhibition of ribosomal peptide synthesis with an IC50
of 0.23 (± 0.01) µM reflecting the improved structural
fit of this molecule. This is significantly better than e. g.
tilmicosin with an IC50 of 0.36 (± 0.02) µM.
Macrolides�comprise�a�large�family�of�naturally-occurring�
compounds�that�exhibit�a�range�of�therapeutic�properties,�
the�best-characterized�and�presently�most�important�of�
which�is�their�antimicrobial�activity��From�a�chemical�stand-
point,�macrolides�are�made�up�of�a�macrolactone�ring,�
consisting�of�12�to�16�atoms,�which�is�substituted�with�up�
to�three�sugar�moieties��The�most�successful�macrolides�
in�hospitals�and�in�the�field�are�chemical�derivatives�of�their�
natural�progenitors�with�improved�stability,�efficacy,�safety�
and�convenience��The�latest�of�these�is�tildipirosin,�a�new�
chemical�entity�(NCE)�for�the�management�of�bovine��
and�swine�respiratory�diseases��Tildipirosin�has�been�shown�
to�be�particularly�efficacious�against�respiratory�tract�
infections�caused�by�the�Gram-negative�bacterial�pathogens�
Mannheimia haemolytica,�Pasteurella multocida,�Histophi-
lus somni,�Bordetella bronchiseptica,�Actinobacillus pleuro-
pneumoniae and�Haemophilus parasuis��
Tildipirosin�is�presently�the�only�tri-basic�16-membered-
ring�macrolide�on�the�market��The�chemical�structure�of�
tildipirosin�has�retained�the�tylonolide�macrolactone�core�
from�the�natural�product�tylosin,�but�differs�in�several�other�
aspects,�the�most�obvious�of�which�are�two�piperidine�
substituents�(Fig��1)��One�piperidine�has�been�added�to�
the�20-position�of�the�tylonolide�ring,�while�the�other�
replaces�the�23-mycinose�sugar��In�addition,�the�mycami-
nose-mycarose�disaccharide�at�the�C5-atom�has�been�
shortened�to�remove�the�mycarose�sugar��The�resultant�
structure�contains�three�basic�centres,�with�pKa�values�of�
9�8,�8�8�and�8�1��The�structure�of�tildipirosin�exhibits�both�
lipophilic�and�hydrophilic�characteristics,�which�vary�
depending�on�the�pH�of�the�environment�and�interaction�
with�membranes�and�proteins��This�imparts�several��
beneficial�properties�including�higher�solubility,�improved�
penetration�into�tissues�and�accumulation�at�the�site�of�
infection�(lung�tissue)��In�addition,�the�effective�penetration�
of�tildipirosin�through�the�outer-membrane�of�Gram-�
negative�bacteria�is�one�of�the�properties�that�make�the�
drug�particularly�effective�against�pathogens�such�as��
M. haemolytica�and�P. multocida�
Figure 1.�Chemical�structure�of�the�tri-basic�16-membered-ring�macrolide�
tildipirosin�
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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While�improved�cell�penetration�is�an�important�character-
istic,�a�successful�antimicrobial�compound�also�needs�to��
be�an�effective�inhibitor�of�bacterial�metabolism��Similar�
to�other�macrolides,�tildipirosin�inhibits�the�process�of�
translation�–�the�synthesis�of�new�peptides�and�proteins�
by�the�bacterial�ribosome�(see�Fig��2)��
Figure 2.�Structure�of�the�bacterial�ribosome��P�indicates�the�peptidyl�trans�-
fer�centre�within�the�50S�unit�of�the�ribosome�where�single�amino�acids�are�
transferred�from�the�tRNA�to�the�growing�peptide�chain��The�red�arrow�
indicates�the�tunnel�used�by�the�nascent�peptide�chain�to�exit�the�ribosome���
The�yellow�circle�highlights�the�binding�site�of�tildipirosin��Adapted�from�
Poehlsgaard,�J��&�Douthwaite,�S��(2005)��Antibiotic�targets�on�the�bacterial�
ribosome��Nature Reviews Microbiology��3:�870�–�881�
Using�an�in vitro�translation�system�with�ribosomes�from�
the�Gram-negative�bacterium�Escherichia coli,�we�showed�
that�50�%�of�protein�synthesis�was�inhibited�by�tildipirosin�
at�0�23�µM�(IC50�=�0�23�±�0�01�µM)�compared�to�another�
16-membered-ring�macrolide�tilmicosin,�which�was�shown�
to�have�an�IC50�value�of�0�36�±�0�02�µM�(see�Table�1)��
16-membered macrolides
IC50* (mean ± standard deviation)
Tildipirosin 0�23�±�0�01�µM
Tilmicosin 0�36�±�0�02�µM
*��Concentration�at�which�50�%�of�protein�synthesis�is�inhibited
Table 1.�Inhibition�of�ribosomal�peptide�synthesis�as�determined�in�
a�coupled�transcription/translation�system�with�ribosome�from�E. coli�
Establishing�that�tildipirosin�was�an�effective�inhibitor�of�
protein�synthesis�led�to�the�questions�of�what�molecular�
interactions�of�tildipirosin�occur�at�its�site�of�inhibition�on�
the�ribosome,�and�whether�these�interactions�differ�from�
those�of�the�other�macrolides��We�addressed�these�issues��
in�two�steps:�First,�the�sites�of�macrolide�drug�contact�were�
mapped�within�the�ribosome�structure�using�chemical�
footprinting,�and�these�data�were�then�used�computa-
tionally�in�molecular�dynamic�simulations�to�visualize�how�
the�drugs�occupy�and�are�oriented�within�their�binding�
site�on�the�ribosome��In�the�case�of�tylosin,�a�high�resolu-
tion�crystal�structure�of�the�drug�bound�to�its�ribosomal�
site�is�available�(PDB:�1K9M)��We�used�this�information�to�
verify�and�to�validate�our�method�of�molecular�dynamic�
simulation��Our�independently�derived�simulation�for�tylosin�
binding�to�the�ribosome�is�compared�with�that�of�the�
crystal�structure�in�Fig��3,�showing�remarkably�consistent�
data�sets��
Figure 3.�The�macrolide�binding�site�in�the�ribosomal�tunnel��Proof�of�prin-
ciple�of�the�experimental�approach��The�crystal�structure�of�tylosin�(salmon,�
PDB:�1K9M)�is�shown�with�the�computationally�minimized�tylosin�binding�
site�superimposed�(red)��Important�nucleotides�from�footprinting�and�the�
crystal�studies�are�numbered�and�shown�in�light�grey;�ribosomal�proteins�
(r-proteins)�that�are�close�to�the�macrolide�site�are�indicated��The�upper�arrow�
points�towards�the�start�of�the�tunnel�at�the�peptidyl�transferase�centre�
(PTC)�and�the�lower�arrow�shows�the�path�taken�by�the�newly�synthesized�
peptide�chain�towards�the�tunnel�exit��The�scale�is�indicated�by�the�
10�ångstrom�bar�
The�footprinting�data�showed�that�the�macrolide�drugs�
tildipirosin,�tilmicosin�and�tylosin�bind�in�the�same�region�
of�Pasteurella�and�E. coli�ribosomes,�the�latter�being�
the�standard�model�for�this�type�of�study��All�macrolides�
interact�with�the�ribosomal�RNA�and�at�nucleotides�
A2058�and�A2059�that�are�located�within�the�tunnel�of�
the�large�ribosomal�subunit��This�observation�was�
expected�from�previous�studies�on�the�mode-of-action�
of�other�macrolide�compounds�(e��g��Poulsen,�et al. 2000��
J Mol Biol,�304:�471�–�481);�however,�at�neighbouring�
nucleotides�there�were�distinguishing�differences�between�
the�contacts�made�by�tildipirosin,�compared�to�the�other�
macrolides�studied�
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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Figure 4.�View�from�another�angle�of�the�computationally�calculated�tylosin�
(red)�and�tildipirosin�binding�(yellow)��The�important�and�distinguishing�con-
stituents�of�the�two�macrolides�are�indicated�
The�two�piperidine�rings�in�tildipirosin�provide�the�potential�
for�unique�interactions�(Fig��4)��Importantly,�the�20-piperi-
dine�is�oriented�into�the�lumen�of�the�ribosome�tunnel�at�a�
location�that�is�not�occupied�by�a�substituent�found�in�any�
other�macrolide��From�this�position,�the�20-piperidine�could�
potentially�block�the�growth�of�the�newly�synthesized�
peptide�chain�as�it�passes�down�the�tunnel��This�model�
is�consistent�with�the�IC50�data,�which�show�a�higher�
potency�of�tildipirosin�(compared�to�tilmicosin)�to�inhibit�
protein�synthesis�on�the�bacterial�ribosome�reflecting�the�
improved�structural�fit�of�this�new�tri-basic�16-membered-
ring�macrolide�to�its�molecular�target�
Notes
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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Notes
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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2.Zuprevo®: Key features of a unique product
Rainer Röpke, Monika Menge, Markus Rose, Andreas Ehinger, Cornelia Bohland, Cornelia Wilhelm, Eva Zschiesche, Susanne Kilp, Wibke Metz and Mark Allan
1
3
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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2
Zuprevo®: Key features of a unique productRainer�Röpke,�Monika�Menge,�Markus�Rose,�Andreas�Ehinger,�Cornelia�Bohland,�Cornelia�Wilhelm,�Eva�Zschiesche,�Susanne�Kilp,�
Wibke�Metz�and�Mark�Allan�
Intervet Innovation GmbH, a member of the MSD Animal Health Group
Abstract
MSD Animal Health has recently been granted
marketing authorization in the European Union
(EU) for Zuprevo® for the treatment and preven-
tion of respiratory disease in cattle. Its active
ingredient, tildipirosin, is the first tri-basic 16-
membered-ring macrolide. The cattle-specific for-
mulation is a ready-to-use, sterile aqueous solution
containing 180 mg of tildipirosin / mL for use as
a single subcutaneous injection (4 mg/kg BW) not
to exceed 10 mL per injection site, equivalent
to 450 kg BW. The unique pharmacokinetics of
Zuprevo® are characterized by rapid absorption
from the injection site and long-lasting high con-
centrations in the lung and bronchial fluid. Mean
maximum plasma concentrations (Cmax) are reached
as early as 23 min (Tmax) post injection and its ter-
minal plasma elimination half life (T1 / 2) is about
9 days. With about 50 L/kg, the volume of distribu-
tion after IV administration is exceptionally large
in comparison to other long-acting macrolides,
while its clearance of 144 mL × h/kg is comparatively
low. The MIC90s of tildipirosin are 1 µg/mL for
M. haemolytica and P. multocida and 4 µg/mL for
H. somni. Tildipirosin concentrations in lung tissue
exceed the H. somni MIC90 for about 16 days and
the MIC90s for M. haemolytica and P. multocida for
at least 28 days. Zuprevo®’s high margin of safety
has been demonstrated in studies exceeding the
label dose of 4 mg tildipirosin / kg BW up to 10 fold.
Furthermore, safety for the consumer is demon-
strated by the high acceptable daily intake of til-
dipirosin (6000 µg / person / day) resulting in a with-
drawal period of 47 days. In conclusion, Zuprevo® is a
novel macrolide with unique pharmacokinetics,
a high margin of safety, and a comparatively short
withdrawal period, offered in a convenient
formulation.
Tildipirosin,�the�active�ingredient�of�Zuprevo®,�is�the�first�
tri-basic�variety�of�16-membered-ring�macrolides�authorized�
and�has�been�developed�exclusively�for�veterinary�use��
The�key�lead�over�previously�developed�compounds�
results�from�the�improved�pharmacokinetics�of�tildipiro-
sin��MSD�Animal�Health�has�recently�been�granted��
marketing�authorization�for�use�of�Zuprevo®�in�bovine�
and�swine�respiratory�disease��The�Zuprevo®�data�pre-
sented�in�this�article�have�been�published�in�the�CVMP�
Assessment�Report�(EMA,�2011)��
Tildipirosin�is�formulated�as�a�ready-to-use,�single-dose�
sterile�aqueous�solution�for�parenteral�injection�as�a�single�
dose�to�combat�respiratory�disease�in�cattle�and�swine���
The�cattle-specific�formulation�containing�180�mg�of�tildip-
irosin�/�mL�guarantees�convenient�dosing�volumes�for�
young�calves�as�well�as�adult�cattle��It�allows�dosing�of�
cattle�up�to�450�kg�BW�with�a�single�injection��The�label�
dose�of�4�mg�tildipirosin�/�kg�BW�is�administered�subcutane-
ously�(SC)�at�2�2�mL�/�100�kg�(1�mL�/�45�kg)�body�weight�(BW)�
for�the�treatment�and�prevention�of�bovine�respiratory��
disease�(BRD)�associated�with�M. haemolytica,�P. multocida�
and�H. somni��Excellent�syringeability�characteristics�based�
on�low�viscosity�at�temperatures�tested�as�low�as�–�10°�C�
also�contribute�to�the�high�convenience�of�Zuprevo®��
Zuprevo®�has�a�high�safety�margin�as�confirmed�in�target-
animal�safety�studies�with�overdosing�up�to�10�times�the�
label�dose�and�is�well�tolerated�following�intravenous�(IV)�
administration�in�cattle��Zuprevo®´s�safety�for�the�con-
sumer�is�demonstrated�by�a�high�acceptable�daily�intake�
(ADI)�of�tildipirosin�(6000�µg�/�person�/�day)�derived�from��
a�comprehensive�package�of�toxicological�and�microbio-
logical�safety�studies;�the�ADI�is�about�10�times�higher�
than�that�of�tulathromycin�(660�µg�/�person�/�day,�Draxxin®:�
EPAR)�and�gamithromycin�(600�µg�/�person�/�day,�Zactran®:�
EPAR)��This�is�reflected�in�a�shorter�withdrawal�period�of�
47�days�for�Zuprevo®�in�comparison�to�other�macrolides�
(49�days,�Draxxin®:�EPAR;�64�days,�Zactran®:�EPAR)��
2
16
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
What�differentiates�Zuprevo®�further�from�other�macro-
lides�is�its�unique�pharmacokinetic�profile��Pharmacoki-
netics�of�Zuprevo®�have�been�investigated�extensively�fol-
lowing�SC�administration�at�different�doses,�in�different�
matrices�and�following�IV�versus�SC�administration��Rapid�
absorption,�extensive�distribution�and�long�elimination�
half-lives�are�the�key�pharmacokinetic�features�of�Zuprevo®��
Mean�maximum�plasma�concentrations�(Cmax)�were�
reached�as�early�as�23�min�(Tmax)�after�SC�administration�
of�4�mg�tildipirosin�/�kg�BW��In�comparison�to�published�
plasma-concentration-versus-time�profiles�of�Draxxin®�
(Nowakowski�et�al�,�2004)�and�Zactran®�(Huang�et�al�,�
2010),�tildipirosin�concentrations�in�plasma�are�higher�and�
more�persistent�as�shown�in�Figures�1�–�3��This�is�reflected�
in�Zuprevo®’s�terminal�plasma�elimination�half�life�(T1�/�2)�
of�about�9�days�which�is�substantially�longer�than�those�
of�Draxxin®�(about�4�days,�Nowakowski�et�al�,�2004),�
Zactran®�(>�2�days,�Huang�et�al�,�2010)�and�Micotil®300�
(about�1�day,�Lombardi�et�al�,�2010)��
Macrolides�are�known�to�accumulate�in�lung�tissue�with�
multi-fold�higher�concentrations�than�in�plasma��With�
about�50�L/kg,�the�volume�of�distribution�of�Zuprevo®�
after�IV�administration�is�exceptionally�large�in�compari-
son�to�those�of�Draxxin®�(about�11�L/kg,�Nowakowski�et�
al�,�2004),�Zactran®�(about�25�L/kg,�Huang�et�al�,�2010)�
and�Micotil®300�(15�3�L/kg,�Lombardi�et�al�,�2010)��At�the�
same�time,�the�clearance�of�Zuprevo®�(144�mL�× h/kg)�is�
lower�than�those�of�Draxxin®�(180�mL × h/kg,�Nowa-
kowski�et�al�,�2004),�Zactran®�(712�mL × h/kg,�Huang�et�
al�,�2010)�and�Micotil®300�(686�mL × h/kg,�Lombardi�et�al�,�
2011)��These�observations�are�confirmed�by�prolonged�
high�lung-tissue�concentrations�of�Zuprevo®�in�compari-
son�to�Draxxin®�and�Zactran®�as�shown�in�Figures�4�–�6��
The�MIC90s�of�tildipirosin�were�determined�against�the�
pathogens�most�commonly�involved�in�the�etiology�of�BRD��
They�were�1�µg/mL�for�M. haemolytica�and�P. multocida�
and�4�µg/mL�for�H. somni��MIC90�values�were�determined�
under�standard�CLSI�testing�conditions�in�epidemio-
logically�unrelated�bovine�isolates�(wild-type)�collected��
in�different�European�countries��Similar�to�other�macro-
lides�such�as�tulathromycin,�gamithromycin�and�tilmico-
sin,�tildipirosin�plasma�concentrations�were�below�MIC90s�
(Nowakowski�et�al�,�2004;�Godinho,�2008;�Huang�et�al�,�
2010;�Modric�et�al�,�1998;�Shryock�et�al�,�1996)��Therefore,�
it�may�be�concluded�that�the�typical�pharmacodynamic�
analyses�appropriate�for�beta-lactam�antibiotics,��
quinolones�and�aminoglycosides�do�not�apply�in�the�
context�of�the�activity�of�the�newer�macrolides��
(Nightingale,�1997)��Other�potential�pharmacodynamic�
properties�of�this�drug�group�include�stimulation�of�
cytokine�pathways�and�enhancement�of�programmed�
cell�death�(apoptosis)�(Lees�et�al�,�2006)��Such�actions�
–�in�addition�to�the�direct�bactericidal�effects�–�may�
exert�additional�or�synergistic�beneficial�effects�in�
infectious�lung�diseases��
Tildipirosin�concentrations�in�lung�tissue�exceed�the��
MIC90�of�H. somni�for�about�16�days�and�the�MIC90s�of�
M. haemolytica�and�P. multocida�for�at�least�28�days�
which�is�much�longer�compared�to�Draxxin®�and�Zactran®�
as�shown�in�Table�1��In�field�trials�in�the�EU,�Zuprevo®’s�
clinical�efficacy�has�been�demonstrated�for�periods�of�
21�days��Predictions�of�efficacy�based�on�lung�homogenate�
concentrations�are�subject�to�scientific�discussions�
(Toutain,�2009)��All�3�respiratory�pathogens�targeted�by�
tildipirosin�attach�to�bronchial�epithelial�cells�and�remain�
extracellular��Therefore,�bronchial�fluid�obtained�from�live,�
non-anesthetized�cattle�at�serial�time�points�was�analysed�
to�demonstrate�bacterial�exposure�to�drug�concentrations�
at�the�site�of�infection��A�method�was�applied�on�the�
basis�of�procedures�described�by�Halstead�et�al��(1991)�
which�allows�the�safe�collection�of�bronchial�fluid��
samples�void�of�blood�and�unaffected�by�dilution�factors��
Bronchial�fluid�samples�were�taken�at�serial�time�points�
post-dose�in�the�same�non-anesthetized�animals��Mean�
bronchial�fluid�tildipirosin�concentrations�exceeded�the�
MIC90s�of�M. haemolytica�and�P. multocida�for�21�days,�
and�approximated�the�MIC90�of�H. somni�for�about�
3�days��The�concentration-versus-time�profiles�of�mean�
tildipirosin�concentrations�in�plasma,�bronchial�fluid�
and�lung�tissue�are�summarized�in�Figure�7��
While�protein�binding�of�tildipirosin�in�bovine�bronchial�
fluid�is�only�30�%,�it�is�acknowledged�that�drug�con-
centrations�measured�in�bronchial�fluid�using�the�method�
described�above�represents�a�conglomerate�of�intra-
cellular�and�extracellular�drug�contents��As�macrolides�are�
known�to�accumulate�within�phagocytic�cells�(Gladue��
et�al�,�1989;�Scorneaux�&�Shryock,�1998),�tildipirosin�con-
centrations�determined�in�bronchial�fluid�also�reflect�that�
portion�of�respiratory�drug�accumulation��Broncho-
alveolar�lavage�(BAL)�has�been�described�by�others�as�a�
17
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
2
method�to�determine�extracellular�drug�concentrations�
in�the�pulmonary�epithelial-lining�fluid�(PELF)�and�intra-
cellular�drug�concentrations�in�the�BAL�cells�(Cox�et�al�,�
2010;�Giguère�et�al�,�2011)��However,�the�analytical��
procedures�remain�a�point�of�discussion�(Zeitlinger�et�
al�,�2005;�Kiem�&�Schentag,�2008),�thus�further�
research�is�required�to�accurately�describe�the�PK�/�PD�
relationships�of�macrolides�used�in�veterinary�medicine�
including�those�of�tildipirosin��
In�conclusion,�Zuprevo®�is�a�novel�16-membered-ring,�
tri-basic�macrolide�with�unique�pharamacokinetics,��
a�high�margin�of�safety,�and�a�comparatively�short�with-
drawal�period,�offered�in�a�convenient�formulation��
References:
1. Cox, S.R., McLaughlin, C., Fielder, A.E., Yancey, M.F., Bowersock, T.L., Garcia-Tapia, D., Bryson, L., Lucas, M.J., Robinson, J.A., Nanjiani, I. & Brown, S.A. (2010) Rapid and prolonged distribution of tulathromycin into lung homogenate and pulmonary epithelial lining fluid of Holstein calves following a single subcutaneous administration of 2.5 mg/kg body weight. International Journal of Applied Research in Veterinary Medicine, 8, 129 – 137.
2. Draxxin: EPAR – Scientific Discussion: http://www.ema.europa.eu/ema/index.jsp?curl=pages/medicines/veterinary/medicines/000077/vet_med_000113.jsp&murl=menus/medicines/medicines.jsp&mid=WC0b01ac058001fa1c
3. EMA (2011) CVMP Assessment Report. Zuprevo (EMEA/V/C/002009). http://www.ema.europa.eu/docs/en_GB/document_library/EPAR_-_Public_assessment_report/veterinary/002009/WC500106577.pdf
4. Halstead, S.L., Walker, R.D., Baker, J.C., Holland, R.E., Stein, G.E. & Hauptman, J.G. (1991) Pharmacokinetic evaluation of ceftiofur in serum, tissue chamber fluid and bronchial secretions from healthy beef-bred calves. Canadian Journal of Veterinary Research, 56: 269 – 274.
5. Huang, R.A., Letendre, L.T., Banav, N., Fischer, J. & Somerville, B. (2010) Pharmacokinetics of gamithromycin in cattle with comparison of plasma and lung tissue concentrations and plasma antibacterial activity. Journal of Veterinary Pharmacology and Therapeutics, 33, 227 – 237.
6. Giguère, S., Huang, R., Malinski, T.J., Dorr, P.M., Tessman, R.K. & Sommerville, B.A. (2011) Disposition of gamithromycin in plasma, pulmonary epithelial lining fluid, bronchoalveolar cells, and lung tissue in cattle. American Journal of Veterinary Research, 72, 326 – 330.
7. Gladue, R.P., Bright, G.M., Isaacson, R.E. & Newborg, M.F. (1989) In vitro and in vivo uptake of azithromycin (CP-62,993) by phagocytic cells: possible mechanism of delivery and release at sites of infection. Antimicrobial Agents Chemotherapy, 33, 277 – 282.
8. Godinho, K.S. (2008) Susceptibility testing of tulathromycin: Interpretative breakpoints and susceptibility of field isolates. Veterinary Microbiology, 129, 426 – 432.
9. Kiem, S. & Schentag, J.J. (2008) Interpretation of antibiotic concentration ratios measured in epithelial lining fluid. Antimicrobial Agents Chemotherapy, 52, 24 – 36.
10. Lees, P., Concordet, D., Aliabadi, F.S. & Toutain, P.L. (2006). Drug selection and optimization of dosage schedules to minimize antimicrobial resistance. In Antimicrobial Resistance in Bacteria of Animal Origin. Ed Aarestrup, F.M., pp. 49 – 60. ASM Press, Washington, D.C., USA.
11. Lombardi, K.R., Portillo, T., Hassfurther, R. & Hunter, R.P. (2011) Pharmacokinetics of timicosin in beef cattle following intravenous and subcutaneous administration. Journal of Veterinary Pharmacology and Therapeutics, doi:10.1111/j.1365 – 2885.2011.01268.x.
12. Nightingale, C.H. (1997) Pharmacokinetics and pharmacodynamics of newer macrolides. Pediatric Infectious Disease Journal, 16, 438 – 443.
13. Nowakowski, M.A., Inskeep, P.B., Risk, J.E., Skogerboe, T.L., Benchaoui, H.A., Meinert, T.R., Sherington, J. & Sunderland, S.J. (2004) Pharmacokinetics and lung tissue concentrations of tulathromycin, a novel triamilide antibiotic in cattle. Veterinary Therapeutics, 5, 60 – 74.
14. Scorneaux, B. & Shryock, T.R. (1999) Intracellular accumulation, subcellular distribution, and efflux of tilmicosin in bovine mammary, blood, and lung cells. Journal of Dairy Science. 82, 1202 – 1212.
15. Shryock, T.R., White, D.W., Staples, J.M. & Werner, C.S. (1996) Minimum inhibitory concentration breakpoints and disk diffusion inhibitory zone interpretive criteria for tilmicosin susceptibility testing against Pasteurella spp. associated with bovine respiratory disease. Journal of Veterinary Diagnostic Investigations, 8, 337 – 344.
16. Toutain, P.L. (2009) PK/PD approach for antibiotics: tissue or blood drug levels to predict antibiotic efficacy. Journal of Veterinary Pharmacology and Therapeutics, 32 (Suppl. 1), 19 – 21.
17. Zeitlinger, M., Mueller, M. & Joukhadar, C. (2005) Lung microdialysis – a powerful tool for the determination of exogenous and endogenous compounds in the lower respiratory tract (mini-review). The AAPS Journal, 7, E600 – E608.
18. Zactran: EPAR – Summary for the public: http://www.ema.europa.eu/ema/index.jsp?curl=pages/medicines/veterinary/medicines/000129/vet_med_000198.jsp&murl=menus/medicines/medicines.jsp&mid=WC0b01ac058001fa1c
2
18
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
Plasma concentrations
0.001
0.01
0.1
1
0.001
0.01
0.1
1
0.001
0.01
0.1
1
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
Time (Days)
Co
nce
ntr
ati
on
[µ
g/m
L]
Plasma concentrations
Plasma concentrations
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
Time (Days)
Co
nce
ntr
atio
n [
µg
/mL]
Co
nce
ntr
atio
n [
µg
/mL]
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
Time (Days)
Zuprevo®
Zactran®
Draxxin®
Plasma concentrations
0.001
0.01
0.1
1
0.001
0.01
0.1
1
0.001
0.01
0.1
1
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
Time (Days)
Co
nce
ntr
ati
on
[µ
g/m
L]
Plasma concentrations
Plasma concentrations
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
Time (Days)
Co
nce
ntr
atio
n [
µg
/mL]
Co
nce
ntr
atio
n [
µg
/mL]
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
Time (Days)
Zuprevo®
Zactran®
Draxxin®
Plasma concentrations
0.001
0.01
0.1
1
0.001
0.01
0.1
1
0.001
0.01
0.1
1
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
Time (Days)
Co
nce
ntr
ati
on
[µ
g/m
L]
Plasma concentrations
Plasma concentrations
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
Time (Days)
Co
nce
ntr
atio
n [
µg
/mL]
Co
nce
ntr
atio
n [
µg
/mL]
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
Time (Days)
Zuprevo®
Zactran®
Draxxin®
Time (Days)
Time (Days)
Time (Days)
Co
nce
ntr
atio
n [
µg
/mL]
Consolidated plasma concentrations
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
0.001
0.01
0.1
1
0.001
0.01
0.1
1
10
100
0 1 2 3 4 5 6 7 8 9 10111213 14151617 18192021 22232425 262728
0.001
0.01
0.1
1
10
100
0 1 2 3 4 5 6 7 8 9 10111213141516171819202122232425262728
Co
nce
ntr
ati
on
[µ
g/g
]
Lung concentrations
Lung concentrations
Co
nce
ntr
atio
n [
µg
/g]
Zuprevo®
Zactran®
Zuprevo®
Draxxin®
Draxxin®
Time (Days)
Time (Days)
Time (Days)
Co
nce
ntr
atio
n [
µg
/mL]
Consolidated plasma concentrations
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
0.001
0.01
0.1
1
0.001
0.01
0.1
1
10
100
0 1 2 3 4 5 6 7 8 9 10111213 14151617 18192021 22232425 262728
0.001
0.01
0.1
1
10
100
0 1 2 3 4 5 6 7 8 9 10111213141516171819202122232425262728
Co
nce
ntr
ati
on
[µ
g/g
]
Lung concentrations
Lung concentrations
Co
nce
ntr
atio
n [
µg
/g]
Zuprevo®
Zactran®
Zuprevo®
Draxxin®
Draxxin®
Time (Days)
Time (Days)
0.001
0.01
0.1
1
10
100
0 1 2 3 4 5 6 7 8 9 10111213141516171819202122232425262728
Consolidated lung concentrations
0.001
0.01
0.1
1
10
100
0 1 2 3 4 5 6 7 8 9 10111213 14151617 18192021 22232425 262728
Co
nce
ntr
ati
on
[µ
g/g
]
1 µg/mL
Co
nce
ntr
atio
n [
µg
/g]
Lung concentrations
Zuprevo®
Zactran®
Draxxin®
Zactran®
Tables and Figures
Figure 1.�Plasma�concentration-time�curve�of�tildipirosin�
(Zuprevo®)�after�SC�administration�of�the�product�label�dose�
DRAXXIN®:�Nowakowski�et�al�(2004)�Veterinary Therapeutics,�5,�60�–�74
DRAXXIN®:�Nowakowski�et�al�(2004)�Veterinary Therapeutics,�5,�60�–�74
ZACTRAN®:�Huang�et�al�(2010)�Journal of Veterinary Pharmacology and Therapeutics,�33,�227�–�237;�Giguère�et�al�(2011)�American Journal of Veterinary Research,�72,�326�–�330
ZACTRAN®:�Huang�et�al�(2010)�Journal of Veterinary Pharmacology and Therapeutics,�33,�227�–�237�
Figure 2.�Plasma�concentration-time�curve�of�tulathromycin�
(Draxxin®)�after�SC�administration�of�the�product�label�dose��
Figure 3.�Plasma�concentration-time�curve�of�
gamithromycin�(Zactran®)�after�SC�administration�
of�the�product�label�dose�
Figure 4.�Lung�tissue�concentration-time�curve�of�
tildipirosin�(Zuprevo®)�after�SC�administration�of�the�
product�label�dose�
Figure 5.�Lung�tissue�concentration-time�curve�of�
tulathromycin�(Draxxin®)�after�SC�administration�of�the�
product�label�dose�
Figure 6.�Lung�tissue�concentration-time�curve�
of�gamithromycin�(Zactran®)�after�SC�administration�
of�the�product�label�dose�
19
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
2
Time (Days)
0 3 6 9 12 15 18 21 24 27 30
µg
Tild
ipir
osi
n/m
L o
r µ
g/g
0.001
0.01
0.1
1
10
100
Blood plasma
Bronchial fluid (in vivo)
Lung tissue
Figure 7.�Consolidated�concentration-time�curves�for�tildipirosin�(Zuprevo®)�in�lung�tissue,�
bronchial�fluid�(�in vivo)�and�plasma�after�SC�administration�of�the�product�label�dose�
Table 1.�Consolidated�times�(days)�above�MIC90�of�M. haemolytica,�P. multocida�and�
H. somni�for�tildipirosin�(Zuprevo®),�tulathromycin�(Draxxin®)�and�gamithromycin�
(Zactran®)�lung�tissue�concentrations�after�SC�administration�of�the�product�label�dose�
Days of drug concentrations in lung tissue above MIC90
M. haemolytica�(MIC90)
P. multocida�(MIC90)
H. somni�(MIC90)
ZUPREVO® >�28� (1�µg/mL) >�28� (1�µg/mL) ~�16� (4�µg/mL)
DRAXXIN® ~�10a� (2�µg/mLb) >�15a,�d�(1�µg/mLb) ~�0�5e� (4�µg/mLb)
ZACTRAN® ~�15c� (0�5�µg/mLc) ~�12c� (1�µg/mLc) ~�12c� (1�µg/mLc)
a�Nowakowski�et�al�(2004)�Veterinary�Therapeutics,�5,�60�–�74b�Godinho�(2008)�Veterinary�Microbiology,�129,�426�–�432c�Giguère�et�al�(2011)�American�Journal�of�Veterinary�Research,�72,�326�–�330d�Cox�et�al�(2010)�International�Journal�of�Applied�Veterinary�Medicine,�8,�129�–�137e��calculated�using�WinNonlin®�software�with�data�derived�from�Nowakowski�et�al�(2004)�Veterinary�Therapeutics,�5,�60�–�74
2
20
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
2
Notes
21
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
3
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
22
1
2
4
3.Preventive efficacy of Zuprevo® in a Mannheimia haemolytica challenge study of bovine respiratory disease
Markus Rose, Wibke Metz, Joachim Ullrich, Eva Zschiesche, Silke Sczesny, Mark Allan and Rainer Röpke
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
23
3
Preventive efficacy of Zuprevo® in a Mannheimia haemolytica challenge study of bovine respiratory diseaseMarkus�Rose,�Wibke�Metz,�Joachim�Ullrich,�Eva�Zschiesche,�Silke�Sczesny,�Mark�Allan�and�Rainer�Röpke
Intervet Innovation GmbH, a member of the MSD Animal Health Group
Abstract
Preventive antimicrobial treatment is an important
element in herd health programs aimed at minimiz-
ing the incidence and costs associated with Bovine
Respiratory Disease (BRD). MSD Animal Health has
recently been granted marketing authorization in
the European Union for Zuprevo® for the treatment
and prevention of respiratory disease in cattle. Its
active ingredient, tildipirosin, the first tri-basic
16-membered-ring macrolide is characterized by rapid
absorption from the subcutaneous injection site and
long-lasting high lung and bronchial fluid concen-
trations. These pharmacokinetic properties make
Zuprevo® an optimal choice for the prevention of
BRD as demonstrated in a Mannheimia haemolytica
challenge study. In this study, tildipirosin (4 mg/kg,
Zuprevo®) was shown to prevent mortality, and was
superior in terms of clinical symptoms and bacterial
counts in lung tissue compared to saline and tulath-
romycin (2.5 mg/kg, Draxxin®).
The role of preventive treatment in bovine respiratory disease (BRD) management
BRD�remains�very�important�to�the�modern�cattle�indus-
try�given�its�high�incidence�in�herds�leading�to�substantial�
economic�losses�through�the�reduced�performance�of�the�
affected�cattle�in�addition�to�treatment�costs�and�mor-
tality�(Babcock�et�al��2010;�Schneider�et�al��2009)��BRD�
outbreaks�most�frequently�occur�following�transportation�
and�commingling�stress�in�young,�newly�weaned�calves,�
often�originating�from�many�different�farms�and�with�
unknown�health�history�or�immune�status�(Step�et�al��
2008;�Taylor�et�al��2010)��The�incidence�of�BRD�in�these�
calves�peaks�within�the�first�few�weeks�after�commin-
gling�(Edwards�2010),�but�BRD�infection�rates�can�remain�
high�until�approximately�80�days�on�feed�(Snowder�et�al��
2006)��Since�commingling�of�calves�is�a�common�manage-
ment�practice,�total�eradication�of�BRD�pathogens�is�
unlikely�(Snowder�et�al��2006)�
BRD�is�a�multi-factorial�disease�caused�by�both�infectious�
and�non-infectious�factors�(Lekeux�1995;�Stoeber�2002;�
Edwards�2010;�Taylor�et�al��2010):
•� �viral�pathogens�(BRSV,�PI-3,�BHV-1,�BVD�/�MD)
•� �bacterial�pathogens�(M. haemolytica, P. multocida,
H. somni, M. bovis)
•� �exo-�and�endogenic�factors�(e��g��management,�
physiology,�environment)
Viral�pathogens�frequently�play�an�initial�role,�predisposing�
cattle�to�secondary�bacterial�pathogens��However,�
M. haemolytica�is�also�known�to�be�a�primary�pathogen�
and�has�traditionally�been�the�most�commonly�isolated�
bacterial�pathogen�(Taylor�et�al��2010)��The�bacterial�
involvement�usually�turns�the�disease�into�a�fibrinous��
or�suppurative�bronchopneumonia�with�clinical�signs�of�
pronounced�dyspnoea�and�fever,�coughing,�nasal�dis-
charge�and�inappetence,�associated�with�moderate�to�
severe�depression�(Stoeber�2002)��However�at�slaughter,�
lung�lesions�may�occur�in�the�majority�of�untreated�cattle�
indicating�also�the�importance�of�subclinically�infected�
animals�and�challenges�with�the�visual�detection�of�BRD�
(Edwards�2010)��
Prevention�is�an�important�element�in�herd�health�pro-
grams�aimed�at�minimizing�the�incidence�and�costs�asso-
ciated�with�BRD�morbidity�and�mortality�(Edwards�2010)�
In�the�EU,�it�is�a�regulatory�requirement�that�the�presence�
3
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
of�the�disease�should�be�confirmed�in�the�herd�before�
preventive�treatment�is�initiated��E��g��a�percentage�of��
the�cattle�in�the�herd�already�present�clinical�signs�of�BRD�
whereas�the�others�show�no�overt�disease�but�may�
already�be�subclinically�infected��In�this�regard,�in�a�recent�
investigation�measuring�body�temperature�by�reticulo-
rumen�temperature�boluses,�fever�episodes�began�4�to�
177�hours�before�treatment�upon�clinical�signs�of�BRD�
(Timsit�et�al��2010)��Therefore,�to�prevent�irreversible�lung�
damage�and�severe�economic�losses,�early�antimicrobial�
treatment�providing�rapid�onset�of�action�and�a�long�
duration�of�activity�is�crucial�to�cover�the�critical�early�fat-
tening�period�following�transportation�and�commingling,�
and�to�avoid�further�contracting�of�the�disease�in�the�herd��
Tildipirosin,�the�active�ingredient�of�Zuprevo®,�is�a�novel�
16-membered-ring,�tri-basic�macrolide�licensed�for�both�
the�treatment�and�prevention�of�BRD��It�is�characterized��
by�rapid�absorption�from�the�subcutaneous�injection�site�
and�long-lasting�high�concentrations�at�the�site�of�in�-
fection�(EMA�2011)��These�pharmacokinetic�features�make�
Zuprevo®�an�optimal�choice�for�the�prevention�of�BRD�
where�it�is�crucial�that�antibiotic�therapy�acts�rapidly�and�
provides�cover�over�a�sustained�period�of�time�to�avoid��
further�spread�of�the�disease�in�the�herd��The�principle�of�
preventive�efficacy�of�Zuprevo®�was�demonstrated�
in�a�Mannheimia haemolytica�challenge�model�of�BRD�
Mannheimia haemolytica challenge study
Materials and methods:�In�dose-finding�experiments,�
18�healthy�male�Holstein�black�pied�calves�were�allocated�
to�three�groups�of�six�animals�each��On�Day�0,�animals�
received�the�following�single�subcutaneous�(SC)�injection:�
•� Group 1:� Zuprevo®,�4�mg�tildipirosin�/�kg�BW�
•� �Group 2:� Draxxin®,�2�5�mg�tulathromycin�/�kg�BW�
•� �Group 3:� Saline,�2�mL�/�100�kg�BW��
Based�on�a�well-established�study�model�(Shoo,�1989),�
five�days�after�treatment�(Day�5)�all�animals�were�chal-
lenged�by�intratracheal�inoculation�of�approximately�1�–�2�
x�109�colony�forming�units�(CFU)�of�M. haemolytica�7�/�2�
Serotype�A�1�(tildipirosin�MIC�of�0�25�µg/mL)�(Figure�1)��
This�pathogenic�strain�provided�by�the�Moredun�Research�
Institute,�Scotland,�originated�from�pneumonic�pasteurel-
losis�and�was�successfully�used�in�other�challenge�studies��
All�animals�were�handled�and�treated�in�accordance�with�
German�animal�welfare�regulations�
Clinical�variables�(body�temperature,�general�behaviour,�
appetite,�and�rate�and�quality�of�respiration)�were�moni-
tored�daily�before�and�twice�daily�after�challenge�by�per-
sonnel�masked�to�treatment��Blinded�necropsy�was�
scheduled�three�days�after�infection�(Day�8)��Lungs�were�
evaluated�for�lung�consolidation�(%)�and�lung-body�
weight�ratios�(%)��Bacteriological�examinations�were�
performed�on�bronchial�swabs�and�lung�tissue�(limit��
of�quantification�[LOQ]�103�CFU�/�g)��
Statistical�analysis�was�performed�by�means�of�the�soft-
ware�package�SAS®��Treatment�unit�and�statistical�unit�
was�the�individual�animal��The�course�of�the�clinical�
examination�parameters�was�graphically�displayed�and�
statistically�analysed��Treatment�groups�were�compared�
pair-wise�as�contrasts�in�a�repeated�measures�analysis�of�
variance,�with�time�and�treatment-time-interaction�as��
the�within-subject�effects�and�treatment�as�the�between-
subject�effect��To�ensure�a�multiple�level�of�significance��
of�α�=�0�05,�the�local�level�of�significance�for�treatment�
effects�was�set�to�α�=�0�005�(Bonferroni�adjustment)��
A�comparison�of�treatment�groups�was�carried�out�for�the�
post�mortem�lung�examination�parameters��Pair-wise�
comparisons�between�treatment�groups�used�a�multiple-
comparison�procedure�to�detect�significant�differences��
The�Ryan-Einot-Gabriel-Welsch�Multiple�Range�Test�was�
used�as�the�multiple-stage�test�which�controlled�the�
maximum�experiment-wise�error�rate�(α�=�0�05)�
Clinical observations:�The�course�of�disease�in�ani-
mals�treated�with�saline�and�Draxxin®�reflects�the�severe�
challenge�by�M. haemolytica�in�this�study��Five�of�the�six�
animals�from�group 3�(saline)�and�one�animal�from�
group 2�(Draxxin®)�died�or�were�euthanized�prior�to�
scheduled�necropsy�for�animal-welfare�reasons�whereas�
no�animal�died�in�group 1�(Zuprevo®)�(Figure�2;�Figure�3)��
General�behaviour,�appetite�and�respiration�scores�(Fig-
ures�4�–�6)�indicated�signs�of�clinical�respiratory�disease�in�
groups 2 and 3�(Draxxin®�and�saline)�after�challenge�until�
the�end�of�the�experimental�phase;�in�contrast,�scores��
in�group 1�(Zuprevo®)�peaked�on�the�day�of�challenge�or�
the�next�day�and�rapidly�decreased�to�normal��Body�
temperature�in�group 1�(Zuprevo®)�increased�slightly�after�
the�challenge;�in�groups 2 and 3�(Draxxin®�and�saline)�
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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the�increase�of�body�temperature�was�higher�(Figure�7)��
Feed�intake�after�infection�was�absent�or�reduced�in�
groups 2�and�3�(Draxxin®�and�saline)��In�group 1�(Zuprevo®),�
the�complete�daily�ration�was�consumed�from�one�day�
after�infection�until�the�end�of�the�study��Body�temperature,�
general�behaviour,�appetite�and�respiration�quality�
scores�were�significantly�lower�in�group 1�(Zuprevo®)�com-
pared�to�groups 2�and�3�(Draxxin®�and�saline)��These�
clinical�variables�were�not�significantly�different�between�
groups 2 and 3 (Draxxin®�and�saline)��
Pathology:�Figure�8�shows�an�example�from�group 3�
(saline�treated�controls)�of�a�lung�severely�affected�by�
bronchopneumonia��Lung�consolidation�(Figure�9)�and�
lung-body�weight�ratios�(Figure�10)�of�group 1�(Zuprevo®)�
were�significantly�lower�than�those�of�groups 2 and 3�
(Draxxin®�and�saline)��No�significant�difference�was�
detected�between�group 2 and 3�(Draxxin®�and�saline)�
Bacteriology:�In bronchial swabs�taken�at�necropsy,�
no�M. haemolytica�growth�was�detected�in�any�from�
the�cattle�treated�with�Zuprevo®�(n�=�0�/�6)��Substantial�
growth�of�M. haemolytica�was�detected�in�bronchial�
swabs�from�all�cattle�in�the�saline-treated�group 3�
(n�=�6�/�6)�and�all�except�for�one�animal�in�the�Draxxin®-
treated�group 2�(n�=�5�/�6)�(Table�1)��
In lung-tissue samples,�growth�of�M. haemolytica�was�
below�the�LOQ�of�103�CFU/g�(n�=�3�/�6)�or�comparatively�
low�(n�=�3�/�6,�105�–�107�CFU/g)�in�cattle�treated�with�Zuprevo®��
Substantially�higher�growth�was�observed�in�lung�tissue��
of�all�animals�in�the�saline-treated�group 3�(n�=�6�/�6;�
109�–�1010�CFU/g)�and�the�Draxxin®-treated�group 3�
(n�=�6�/�6;�107�–�109�CFU/g)�(Table�1)��
Clinical,�pathological�and�bacteriological�investigations�
showed�that�the�preventive�efficacy�of�Zuprevo®�in�this�
severe�M. haemolytica�challenge�model�of�BRD�was�superior�
in�comparison�to�saline-treated�controls�and�Draxxin®��
Preventive�treatment�with�Zuprevo®�provided�superior�
reduction�of�pulmonary�tissue�bacterial�counts��The�
reduction�of�M. haemolytica�to�below�the�LOQ�in�lung�
tissue�in�50�%�of�the�animals�and�in�bronchial�fluid��
in�100�%�of�the�animals�indicates�bactericidal�action��
of�Zuprevo®�(active�ingredient�tildipirosin)�against�
M. haemolytica�and�confirms�its�long�duration�of�action�
at�high�concentrations�at�the�site�of�infection�
References:
1. Babcock AH et al. (2010). Temporal distributions of respiratory disease events within cohorts of feedlot cattle and associations with cattle health and performance indices. Prev Vet Med 97, 198 – 219.
2. Step DL et al. (2008). Effects of commingling beef calves from different sources and weaning protocols during a forty-two-day receiving period on performance and bovine respiratory disease. J Anim Sci 86, 3146 – 3158.
3. Edwards TA (2010). Control methods for bovine respiratory disease for feedlot cattle. Vet Clin Food Anim 26, 273 – 284.
4. Schneider MJ et al. (2009). An evaluation of bovine respiratory disease complex in feedlot cattle: Impact on performance and carcass traits using treatment records and lung lesions scores. J Anim Sci 87, 1821 – 1827.
5. Taylor JD et al. (2010). The epidemiology of bovine respiratory disease: What is the evidence for predisposing factors? Can Vet J 51, 1095 – 1102.
6. Snowder GD et al. (2006). Bovine respiratory disease in feedlot cattle: Environmental, genetic, and economic factors. J Anim Sci 84, 1999 – 2008.
7. Lekeux P (1995). Bovine respiratory disease complex: A European perspective. Bovine Practitioner 29, 71 – 75.
8. Stoeber M (2002). Krankheiten der Atmungsorgane, des Zwerchfells und der Brustwand: Enzootische Bronchopneumonie. In: G Dirksen, H-D Gründer, M Stoeber (eds.) Innere Medizin und Chirurgie des Rindes, 4th ed, Blackwell, pp. 310 – 322.
9. Timsit E et al. (2010). Early detection of bovine respiratory disease in young bulls using reticulo-rumen temperature boluses. Vet J doi:10.1016/j.tvjl.2010.09.012.
10. EMA (2011). CVMP Assessment Report. Zuprevo (EMEA/V/C/002009). http://www.ema.europa.eu/docs/en_GB/document_library/EPAR_-_Public_assessment_report/veterinary/002009/WC500106577.pdf
11. Shoo MK (1989). Experimental bovine pneumonic pasteurellosis: A review. Vet Record 124, 141 – 144.
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
Necropsy &Bacteriology
0 7 85 63 421
Treatment
Studyday
Infection
0
1
2
3
4
5
6
0 1 2 3 4 5 6 7 8Time (days)
Mo
rtal
itie
s /
no
n-s
che
du
led
eu
than
asia
Infection
Group 3: Saline
Group 2: Draxxin®
Group 1: Zuprevo®
Group 1: Zuprevo®
Group 2: Draxxin®
Group 3: Saline
0
1
2
3
4
5
6
Mo
rtal
itie
s /
no
n-s
ched
ule
deu
than
asia
Group 1: Zuprevo®
Group 2: Draxxin®
Group 3: Saline
Group 1: Zuprevo®
Group 2: Draxxin®
Group 3: Saline
0.0
0.5
1.0
1.5
2.0
2.5
3.0
0 1 2 3 4 5 6 7 8
Time (days)
Sco
re
Infection
0.0
0.4
0.8
1.2
1.6
2.0
2.4
2.8
3.2
0 1 2 3 4 5 6 7 8Time (days)
Sco
re
Infection
Group 1: Zuprevo®
Group 2: Draxxin®
Group 3: Saline
Tables and Figures
Figure 1.�Study�Schedule
Monitoring�of�general�behaviour,�body�temperature,�appetite,�rate�&�quality�of�respiration*
Infection�by�intratracheal�inoculation�of�~1�–�2�x�109�colony�forming�units�(CFU)�M. haemolytica�7�/�2�Serotype�A�1�(tildipirosin�MIC�of�0�25�µg/mL)
*�Personnel�masked�to�treatment
Score 0:�normal;�Score 1:�slight�depression;�Score 2:�moderate�depression;�Score 3:�severe�depression;�Score 4:�severe�prostration/recumbency�(animals�were�euthanized)Group�1�significantly�different�from�Groups�2�and�3�
Score 0:�normal�appetite�(animal�directly�approaches�the�feed);�Score 1:�appetite�slightly�restricted;�Score 2:�appetite�restricted;�Score 3:�no�appetite�Group�1�significantly�different�from�Groups�2�and�3�
Figure 2.�Mortality�
Figure 3.�Mortality�(summary)
Figure 4.�General�behavior�scores�(means;�Day�0�–�Day�8)�
Figure 5.�Appetite�scores�(means;�Day�0�–�Day�8)�
27
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
3
05
1015
20253035404550
3 days after infection
3 days after infection
*
*
Lun
gco
nso
lidat
ion
(%)
1
1,2
1,4
1,6
1,8
2
2,2
Lun
g-b
od
yw
eig
ht
rati
o(%
)
Group 1: Zuprevo®
Group 2: Draxxin®
Group 3: Saline
Group 1: Zuprevo®
Group 2: Draxxin®
Group 3: Saline
05
1015
20253035404550
3 days after infection
3 days after infection
*
*
Lun
gco
nso
lidat
ion
(%)
1
1,2
1,4
1,6
1,8
2
2,2
Lun
g-b
od
yw
eig
ht
rati
o(%
)
Group 1: Zuprevo®
Group 2: Draxxin®
Group 3: Saline
Group 1: Zuprevo®
Group 2: Draxxin®
Group 3: Saline
0.0
0.4
0.8
1.2
1.6
2.0
2.4
2.8
3.2
0 1 2 3 4 5 6 7 8Time (days)
Sco
re
Infection
Group 1: Zuprevo®
Group 2: Draxxin®
Group 3: Saline
37
38
39
40
41
0 1 2 3 4 5 6 7 8Time (days)
Bo
dy
tem
per
atu
re (°
C)
Infection
Group 1: Zuprevo®
Group 2: Draxxin®
Group 3: Saline
Figure 6.�Respiration�quality�scores�(means;�Day�0�–�Day�8)�
Score 0:�normal;�Score 1:�slight�dyspnea;�Score 2:�moderate�dyspnea;�Score 3:�severe�dyspneaGroup�1�significantly�different�from�Groups�2�and�3
Group�1�significantly�different�from�Groups�2�and�3 *�Group�1�significantly�different�from�Groups�2�and�3
*�Group�1�significantly�different�from�Groups�2�and�3
Figure 7.�Body�temperature�(means;�Day�0�–�Day�8)
Figure 8:�Lung�severely�affected�by�bronchopneumonia�
(example�from�Group�3;�saline�treated�controls)
Figure 9.�Lung�consolidation�(means;�%)�
Figure 10.�Lung-body�weight�ratio�(means;�%)�
Table 1.�Bronchial-fluid�and�lung-tissue�bacteriology�
Matrix Treatment Growth�of�M. haemolytica*
Bron-chial fluid
Group�1:�Zuprevo® –�(6�/�6)
Group�2:�Draxxin® +++�or�++++�(5�/�6)
Group�3:�Saline ++++�(6�/�6)
Lung tissue
Group�1:�Zuprevo® <�LOQ�(3�/�6);�105�–�107�(3�/�6)
Group�2:�Draxxin® 107�–�109�(6�/�6)
Group�3:�Saline 109�–�1010�(6�/�6)
*��Bronchial�fluid:�“–“�no�growth;�“+++“�pre-confluent�growth;��“++++”�confluent�growth��Lung�tissue:�CFU/g;�limit�of�quantification�(LOQ)�=�103�CFU/g
3
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
Notes
3
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
30
4.Zuprevo®: Efficacy in the treatment of BRD in calves in comparison to Draxxin®
Kerstin E. Müller, Franziska Schmidt, Nadja Rohdich, Eva Zschiesche and Rainer Röpke
1
2
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
31
4
Zuprevo®: Efficacy in the treatment of BRD in calves in comparison to Draxxin®Kerstin�E��Müller1,�Franziska�Schmidt1,�Nadja�Rohdich2,�Eva�Zschiesche2�and�Rainer�Röpke2
1 Clinic for Ruminants and Swine, Faculty of Veterinary Medicine, Freie Universität Berlin, Berlin, Germany
2 Intervet Innovation GmbH, a member of the MSD Animal Health Group
Abstract
Bovine Respiratory Disease (BRD) in calves is still
a main problem in cattle production worldwide.
The multifactorial etiology of BRD poses a challenge
to farmers and veterinarians. Efficacious treatment
of BRD based on adequate diagnostic tools is crucial
to restore and maintain herd health. MSD Animal
Health has recently been granted marketing authori-
zation for Zuprevo® for the treatment and prevention
of respiratory disease in cattle. Its active ingredient,
tildipirosin, is the first tri-basic, 16-membered-ring
macrolide. Tildipirosin is characterized by rapid
absorption from the subcutaneous injection site and
long-lasting high lung and bronchial fluid concen-
trations. These pharmacokinetic properties make
Zuprevo® particularly suited for the treatment of
BRD as demonstrated in a field trial in calves with
BRD. In this study, Zuprevo® was efficacious and
safe in the treatment of BRD over an observation
period of 21 days. The onset of efficacy was most
rapid with Zuprevo®. Overall, efficacy results were sig-
nificantly non-inferior in comparison to Draxxin®.
Bovine Respiratory Disease Complex – etiology, diagnosis and treatment
The�Bovine�Respiratory�Disease�Complex�(BRDC)�is�defined�
as�a�multifactorial�disease�of�the�respiratory�tract�resulting�
from�complex�interactions�between�environmental�and��
host�factors�and�pathogens�(U�S��National�Library�of�
Medicine®)��These�factors�act�as�stressors�that�adversely�
affect�the�immune�system�and�other�host�defence�mecha-
nisms�leading�to�an�enhanced�transmission�of�infectious�
agents��For�calves�several�risk�periods�have�been�identi-
fied�when�the�animals�are�prone�to�develop�respiratory�
diseases:�the�neonatal�period,�the�period�around�weaning�
and�the�period�when�maternal�antibodies�have�vanished��
Beef�and�veal�calves,�in�addition,�experience�extra�risk�
periods�due�to�commingling�of�animals�during�the�first�
ten�days�after�arrival�at�the�fattening�farm�and�whenever�
regrouping�of�animals�takes�place��Besides�these�critical�
periods,�numerous�factors�have�been�identified�that�con-
tribute�to�outbreaks�of�BRDC��These�include�host�factors�
(anatomical�and�functional�weaknesses�of�the�bovine�
respiratory�tract,�genetics),�the�macro-�and�the�microcli-
mate�(temperature,�humidity,�air�velocity,�toxic�gases,�
dust),�management�factors�(management�of�the�neonate,�
hygiene,�nutrition,�health�status�of�the�herd)�and�the�
presence�of�certain�microorganisms�(Figure�1)��Among�
viruses�of�relevance�for�the�BRDC,�the�Bovine�Virus��
Diarrhoea�Virus�(BVDV),�the�Bovine�Respiratory�Syncytial�
Virus�(BRSV)�and�the�Bovine�Herpesvirus�-1�(BoHV-1)�play��
a�major�role�(Thiery,�2007)��The�same�is�true�for�bacteria�
as�Mannheimia haemolytica,�Pasteurella multocida,�Biber-
steinia trehalosi,�Histophilus somni,�Arcanobacterium pyo-
genes�and�mycoplasmata�(M. bovis)�(Griffin�et�al�,�2010)��
Most�microorganisms�are�normal�inhabitants�of�the�bovine�
upper�respiratory�tract��Due�to�certain�strategies�the��
latter�bacteria�are�able�to�prevail�over�other�bacteria�and�
weaken�the�host’s�defence�mechanisms��Several�tech-
niques�are�available�to�sample�the�bovine�respiratory�tract�
in�order�to�detect�microorganisms�participating�in�BRDC�
(Cooper�and�Brodersen,�2010)��Less�invasive�techniques�
include�obtaining�nose�swabs,�sputum,�tracheal�swabs�
and�blood�samples��Trans-tracheal�lavage,�tracheobronchial�
lavage,�broncho�alveolar�lavage�and�lung�biopsy�are�more�
invasive�inter�ventions��The�procedure�of�(trans-)�tracheal�
lavage�is�a�suitable�tool�for�the�detection�of�bacteria�
species�that�participate�in�BRDC�as�the�lower�respiratory�
tract�of�healthy�calves�has�been�shown�to�be�sterile�
4
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
(Fischer�et�al�,�1987;�Heckert�et�al�,�1997)�(Figure�2)��
Calves�suffering�from�BRDC�can�be�assigned�to�four�dif-
ferent�categories�based�on�clinical�findings�(Reinhold,�
2001):�Grade�I�(subclinical)�includes�alert�calves�with�normal�
breathing�sounds�at�auscultation,�Grade�II�(compensated)��
is�characterized�by�the�presence�of�symptoms�of�airway��
disease,�a�short�period�of�fever�and�sustained�appetence,�
Grade�III�(non-compensated)�describes�calves�demon-
strating�dyspnoea,�inappetence,�enhanced�or�even�bronchial�
breathing�sounds)�and�Grade�IV�(irreversible)�includes�calves�
exhibiting�severe�dyspnoea�and�anorexia�and�bronchial�
breathing�sounds�or�even�absent�breathing�sounds�over�
extended�areas�of�the�lung�field�(Table�1)��A�treatment�
advice�is�linked�to�each�of�the�four�grades�(Reinhold,�2001):�
Grade�I�calves�do�not�need�a�specific�treatment,�Grade�II�
demands�an�antibacterial�treatment,�Grade�III�a�combination�
of�antibiotics�and�anti-inflammatory�drugs�preferentially�
non-steroidal-anti-inflammtory�drugs��Grade�IV�(irreversible)�
calves�are�unlikely�to�recover�from�BRDC�due�to�the�
extended�and�irreversible�damage�to�their�lungs�and�for�
this�reason�should�be�euthanized�(Table�1)�
A Field Trial
Zuprevo®�(active�ingredient�tildipirosin)�is�efficacious�in�
the�treatment�of�naturally�acquired�Bovine�Respiratory�
Disease�(BRD)��Previously�performed�field�studies�had�
evaluated�the�primary�efficacy�endpoint�up�to�14�days�after�
treatment;�however,�secondary�efficacy�parameters�in�
those�studies�suggested�an�efficacy�determined�over�a�
21�days�observation�period�
This�study�was�designed�to�confirm�the�efficacy�of�Zuprevo®,�
administered�subcutaneously�at�a�single�dose�of�4�mg��
tildipirosin/kg�body�weight�(BW)�in�the�treatment�of�BRD�
under�field�conditions�over�an�observation�period�of�
21�days��Efficacy�was�compared�to�the�registered�product�
Draxxin®�(active�ingredient�tulathromycin,�2�5�mg/kg�BW)��
The�study�was�conducted�in�accordance�with�Good�Clinical�
Practice,�VICH�GL9�(CVMP/VICH/595/98-FINAL)�
Materials and methods
The�study�had�an�investigator-blinded,�positive-controlled�
design�and�was�conducted�at�one�site�in�Germany��
104�calves�(11�to�56�days�old)�were�included�in�the�study,�
52�per�treatment�group��Animals�showing�the�following�
signs�of�BRD�on�day�0�were�included�in�the�study�(Table�2):
•� Rectal�temperature�≥ 40°�C��
•� Abnormal�respiration�(respiratory�score�≥ 1)��
•� Abnormal�general�attitude�(attitude�score�≥ 1)��
Animals�were�randomly�allocated�to�treatment�with�either�
Zuprevo®�(4�mg�tildipirosin/kg�BW)�or�Draxxin®�(2�5�mg�
tulathromycin/kg�BW)��Treatments�were�administered�as�
a�single�subcutaneous�injection�
Clinical�examinations�were�performed�daily�from�day�0�until�
day�10�and�on�day�21��From�day�0�to�day�2,�examinations�
were�done�three�times�daily�(approximately�6�hours�apart)�
and�once�daily�from�day�3�to�day�10��Between�day�11�
and�day�20,�animals�were�observed�daily�for�general�health��
Detailed�examinations�were�performed�in�case�of�re-occur-
rence�of�BRD�signs��Injection�sites�were�observed�before�
treatment�and�on�days�1,�2,�3,�10,�and�21�after�treatment�
On�day�0,�all�animals�were�sampled�for�bacteriological�
examination�using�the�trans-tracheal�lavage�(TTL)�tech-
nique��On�day�0�and�21,�animal�weights�were�determined�
and�information�on�potential�concomitant�viral�and�myco-
plasma�infections�was�gathered�by�obtaining�paired�
serum�samples�from�all�animals��Haptoglobin�concentra-
tions�were�determined�by�photometric�assay�in�serum�
samples�taken�on�day�0,�1,�2,�3,�10,�and�21��Between�day�
0�and�day�21,�feed�intake�was�monitored�
Animals�were�regarded�as�treatment�failures�and�with-
drawn�from�the�study�if�they�fulfilled�the�following�criteria:
•� Between�day�2�and�day�10:�rectal�temperature�≥ 40°�C�
and�(respiratory�score�≥ 2�or�attitude�score�≥ 2)��
•� Between�day�11�and�day�20:�rectal�temperature�≥ 40°�C�
or�respiratory�score�≥ 2�or�attitude�score�≥ 1��
•� On�day�21:�rectal�temperature�≥ 40°�C�or�respiratory�
score�≥ 1�or�attitude�score�≥ 1��
In�case�of�animal�withdrawal�from�the�study�between�day�0�
and�day�21�due�to�BRD-related�reasons�(failure,�late�failure),�
these�animals�were�re-sampled�for�bacteriological�examina-
tion�using�the�TTL�technique�on�the�day�of�withdrawal�
Necropsy�was�performed�on�6�animals�which�completed�the�
study�successfully�on�day�21,�3�treated�with�Zuprevo®�and�3�
with�Draxxin®��Lung�tissue�was�sampled�for�bacteriological�
examination�and�lung�lesion�scores�were�determined�
33
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
4
Primary�efficacy�criterion�was�the�treatment�success�rate��
on�day�21,�defined�as�the�percentage�of�animals�remaining�
in�the�study�on�day�21�
Results
Long�term�treatment�success�rates�(treatment�success�on�
day�21)�were�72�1�%�(31�of�43)�in�the�Zuprevo®�group�and�
62�2�%�(28�of�45)�in�the�Draxxin®�group�(per-protocol�[PP]�
population)��Short�term�treatment�success�rates�(inter-
mediate�treatment�success�on�day�10)�were�83�7�%�(36�of�
43)�in�the�Zuprevo®�group�and�84�4�%�(38�of�45)�in�the�
Draxxin®�group�(PP)��Long�term�treatment�failure�on�days�
11�–�21�after�short�term�(day�10)�success�(late�failures)�
were�13�9�%�(5�of�36)�in�the�Zuprevo®�group�and�26�3�%�
(10�of�38)�in�the�Draxxin®�group��For�all�3�criteria,�Zuprevo®�
was�non-inferior�to�Draxxin®�(Table�3)��
No�mortality�occurred�in�the�Zuprevo®�group,�but�2�
animals�treated�with�Draxxin®�died�(1�in�the�PP,�1�in�the�
intent-to-treat�[ITT]�population)��
Rectal�temperatures�(Figure�3),�respiratory�and�attitude�
scores�(Figure�4),�and�respiration�frequency�decreased�
markedly�in�both�treatment�groups�(PP)�starting�at�day�1��
Daily�weight�gains�or�lung�scores�did�not�differ�significantly�
between�groups�(PP)��
Before�treatment,�Mannheimia (M.) haemolytica�(18�3�%),�
Pasteurella (P.) multocida�(31�7�%),�and�Histophilus (H.)
somni�(1�0�%)�were�isolated�by�TTL�(ITT)�from�the�respira-
tory�tract�of�the�study�animals��At�withdrawal,�distri-
bution�of�isolates�was�as�follows:�8�3�%�M. haemolytica,�
33�3�%�P. multocida,�and�0�0�%�H. somni�(ITT)��Mycoplasma
( M.) bovis�was�not�isolated;�however,�4�animals�had�con-
comitant�M. bovis�infections�as�detected�by�seroconver-
sion�(ITT)��The�minimum�inhibitory�concentrations�(MIC)�of�
tildipirosin�were�determined�(Table�4)��
Haptoglobin�levels�reflected�the�disease�status�in�both�
treatment�groups�(data�not�shown)��During�the�study,�
feed�intake�did�not�differ�between�both�groups��
Mean�onset�of�efficacy�(i��e��first�treatment�effects�as�shown�
by�rectal�temperature�<�39�5°�C�and�attitude�score�=�0)��
was�26�8�h�in�the�Zuprevo®�group�and�39�4�h�in�the�Draxxin®�
group�(PP)�(Table�5)��
No�pain�reaction�was�noted�on�injection�in�86�5�%�of�
the�animals�in�the�Zuprevo®�group�and�in�65�4�%�in�the�
Draxxin®�group�(ITT)��9�6�%�of�animals�in�the�Zuprevo®�
group�and�13�5�%�in�the�Draxxin®�group�had�injection�site�
reactions�(ITT)��16�adverse�events�(concomitant�diseases�
e��g��otitis,�diarrhoea,�lameness,�omphalitis)�were�reported,�
12�in�the�Zuprevo®�group�and�4�in�the�Draxxin®�group��
All�except�one�(rated�“unknown”)�were�unlikely�related�to�
the�study�medication�(ITT)�
Conclusion
Zuprevo®�administered�subcutaneously�at�a�single�dose�
of�4�mg/kg�BW�was�efficacious�and�safe�in�the�treat-
ment�of�BRD�within�an�observation�period�of�21�days��
The�onset�of�efficacy�was�most�rapid�with�Zuprevo®��
Overall,�efficacy�results�were�significantly�non-inferior�
in�comparison�to�Draxxin®�
References:
1. Cooper, V.L., Brodersen, B.W. (2010): Respiratory Disease Diagnosis of Cattle. The Vet. Clin. North Am. 26(2): 409 – 416.
2. Fischer, W., Amtsberg, G., Luitjens, B. (1987): Vergleichende Untersuchungen zur Keimbesiedlung der Nasen- und Tracheobronchial-schleimhaut bei bronchopneumonisch erkrankten Kälbern und Jungrindern. Tierärztl. Umsch. 42(6): 476 – 480.
3. Griffin, D. Chengappa, M.M., Kuszak, J., McVey, D. (2010): Bacterial Pathogens of the Bovine Respiratory Disease Complex. The Vet. Clin. North Am. 26(2): 381 – 394.
4. Heckert, H.-P., Rohn, M., Hofmann W. (1997): Diagnostische Probenentnahmen bei infektiösen Atemwegserkrankungen der Rinder. Der Praktische Tierarzt 78(12): 1056 – 1065.
5. Reinhold, P. (2001): Untersuchungen zur Bestimmung pulmonaler Funktionen beim Rind. Habilitationsschrift Freie Universität Berlin.
6. Thiry, E. (2007): Clinical Virology of Ruminants. Wolters-Kluwer, France pp. 19 – 54.
4
34
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
microorganisms
macro- and microclimate
host genetics�immune�system�general�condition
management care�for�neonates�crowding�nutrition�(weaning)�loss�of�colostral�protection�hygiene
0.0
0.4
0.8
1.2
1.6
2.0
2.4
Day
0,
Tim
e 1
Day
0,
Tim
e 2
Day
0,
Tim
e 3
Day
1,
Tim
e 1
Day
1,
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e 3
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Tim
e 3
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Study day
Mea
n a
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38,0
38,5
39,0
39,5
40,0
40,5
41,0
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Tim
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Study day
Rec
tal
Tem
per
atu
re [
°C]
Zuprevo®
Draxxin®
Zuprevo®
Draxxin®
0.0
0.4
0.8
1.2
1.6
2.0
2.4
Day
0,
Tim
e 1
Day
0,
Tim
e 2
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Tim
e 3
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1,
Tim
e 1
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Tim
e 2
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e 3
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2,
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e 1
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2,
Tim
e 2
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2,
Tim
e 3
Day
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Day
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10
Study day
Mea
n a
ttit
ud
e s
core
38,0
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Day
0,
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e 1
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0,
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e 2
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e 3
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1,
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e 1
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1,
Tim
e 2
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1,
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e 3
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2,
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e 1
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2,
Tim
e 2
Day
2,
Tim
e 3
Day
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Day
4
Day
5
Day
6
Day
7
Day
8
Day
9
Day
10
Study day
Rec
tal
Tem
per
atu
re [
°C]
Zuprevo®
Draxxin®
Zuprevo®
Draxxin®
Figure 1.�Etiology�of�BRD�
Figure 2.�Trans-tracheal�lavage�(TTL)�technique�
Figure 3.�Course�of�rectal�temperatures�(PP�population)� Figure 4.�Attitude�score�(PP�population)�
Tables and Figures
35
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
4
Table 1.�Severity�of�Bovine�Respiratory�Disease�based�on�clinical�symptoms�and�treatment�advice�(Reinhold�2001)�
Table 3.�Efficacy�results�
Table 2.�Study�schedule�
Grade Description Treatment
I Subclinical�(alert,�normal�breathing�sounds) No�treatment
II Compensated�(symptoms�of�airway�disease,�appetence,�short�period�of�fever) Antibiotics�*
III Non-compensated�(dyspnoea,�inappetence,�enhanced�or�bronchial�breathing�sounds) Antibiotics��and�NSAID�*
IV Irreversible�(severe�dyspnoea,�anorexia,�bronchial�or�even�absent�breathing�sounds) Euthanasia
*�Bronchosecretolytica�on�indication
Parameter Zuprevo® Draxxin®
Long-term�treatment�success�day�21�(Treatment�success�day�21) 72�1�%*� (31�of�43) 62�2�%� (28�of�45)
Short-term�treatment�success�day�10�(Intermediate�success�day�10) 83�7�%*� (36�of�43) 84�4�%� (38�of�45)
Long-term�failure�(days�11�–�21)�after�short-term�(day�10)�success(Late�failure�day�11�–�21)
13�9�%*� (5�of�36) 26�3�%� (10�of�38)
*Significantly�non-inferior�to�Draxxin®
Study day Action Sample Evaluation injection site
Day�0 Clinical�examinationInclusionWeighing
Allocation�to�the�treatment�groups
TTL�and��serum�sampling1
Yes�3
Group A
Treatment�with�Zuprevo®:�4�mg�tildipirosin/kg�BW
Group B
Treatment�with�Draxxin®:�2�5�mg�tulathromycin/kg�BW
Clinical�examination�2�x
Day�1�–�Day�2 Clinical�examination�3�x Clinical�examination�3�x Serum�day�1,�2�2 Yes�3
Day�3�–�Day�10 Clinical�examination�once�daily�(scheduled�examination) Serum�day�3,�10�2 Days�3,�10
Day�11�–�Day�20 General�health�observations,�examination�if�necessary��(unscheduled�examination)
In�case�of�examination
Day�2�–�Day�21 Withdrawal�due�to�BRD TTL
Day�21 Clinical�examination,�weighing Serum�sampling1 Yes
Necropsy�of�3�animals�4 Necropsy�of�3�animals�4 Lung�tissue
End�of�animal�phase
1�Serum�sampling�for�determination�of�antibodies�and�haptoglobin2�Serum�sampling�for�determination�of�haptoglobin3�Once�per�day4�Animals�that�completed�the�study�as�treatment�success
4
36
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
Notes
4
Table 4.�Minimum�inhibitory�concentrations�(MICs)�of�tildipirosin�
Table 5.�Onset�of�efficacy�(PP�population)�
Parameter MIC of tildipirosin [µg/mL]
M. haemolytica (N = 21) P. multocida (N = 42) H. somni (N = 1)
Minimum�MIC� 0�25 0�25 2
Maximum�MIC� 0�5 2 2
Treatment group Onset of efficacy [h]*
Mean Standard deviation Min Max
Zuprevo®�(N�=�31) 26�8 18�9 5�8 96�0
Draxxin®�(N�=�28) 39�4 42�8 10�8 216�0
*First�treatment�effects�as�shown�by�rectal�temperature�<�39�5°�C�and�attitude�score�=�0
37
6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
Notes
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
Notes
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6th European Congress of Bovine Health ManagementZuprevo® – A new paradigm in the treatment and prevention of BRD – September 2011
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