Note: Descriptions are shown in the official language in which they were submitted.
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Treatment of serious infections and septic shock
The invention relates to the use of rhesus theta-defensin 1 (RTD-1 ) for
producing a
medicament for the treatment and/or prophylaxis of patients with serious
infections
(bacteremias) including septic shock.
Sepsis is a severe, life-threatening syndrome resulting from an infection with
bacteria
at the systemic level (bacteremia) and further clinical findings according to
the
internationally valid definition (Madot, I. and Sprung, C.L. (2001 ), Int.
Care Med. 27,
S.3-S.9) which includes inter alia also the systemic inflammatory reaction of
the
body with subsequent organ failure. If a local infection by the pathogens
breaks
through the endogenous barriers (e.g. epithelial, endothelial, blood-brain
barner), a
sepsis may develop from the bacteremia resulting therefrom. The microbes
continuously enter the blood stream, and thus the whole body, from the septic
focus
(e.g. abscess, lung, gastrointestinal tract), which may also remain
unidentified.
Although the defense functions of the immune system impede reproduction of the
microbes which have entered the blood and other immunological organs, they are
usually not completely destroyed. In addition, the attack on the microbes by
the
immune system, and also therapies with certain antibiotics, result in the
release of
bacterial products such as lipopolysaccharide (LPS), lipoteichoic acid (LTA)
and the
like (Nau, R. and Eiffert, H. {2002), Clin. Microbiol. Rev. 15, 95-110).
Sepsis is characterized by fever, hypotension and so-called shock symptoms
(e.g.
shock lung, renal failure, gastrointestinal bleeding; generally referred to as
multiorgan failure). These different symptoms are the clinical signs of
pathophysiological processes brought about by the microbes themselves or their
products, e.g. endotoxins, hemolysins or pyrogens. Further causes which may
also
occur are pathological states such as severe burns, trauma or acute pulmonary
changes with subsequent or simultaneous colonization by bacteria, fungi or
viruses.
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Shock symptoms are also found in these cases, but only in some cases is direct
diagnosis of the bacteria or other pathogens possible.
The release of bacterial products or the bacteria themselves lead to a
response by the
body through the immune system. Immune system factors intrinsic to the patient
are
in these cases both protective and harmful, depending on the concentration,
site of
action and the like. )endogenous factors released as a response of the body to
external
stimuli interact in a complex fashion in the tissues and thus lead in some
circumstances to the pathological state of sepsis. These events are initiated
by
definition through the presence of bacteria and/or the presence of bacterial
products
such as LPS/LTA and others. This leads to the release of, for example, tumor
necrosis factor a, interleukin-1 (IL-1), interleukin-6 (IL-6) and interleukin-
8 (IL-8),
and factors which influence coagulation (e.g. platelet activating factor, PAF)
and
factors which intervene to regulate the resulting inflammatory event (such as,
for
example, prostaglandins, leukotrienes, interleukin-10). An overreation of the
body,
and especially of the immune system, results in the development far from the
orginally protective and infection-repelling response, of the clinical feature
of
bacteremia and subsequently of sepsis (Cohen, J. (2002), Nature 420, 885-891).
Cytokine analyses in septic patients show a significant correlation with
elevation of,
in particular, IL-6, IL-8 and TNF-a and an increased mortality (Rodriguez-
Gaspar M.
et al. (2001), Cytokine 15, 232-236).
Although in principle virtually all microorganisms may induce bacteremia and
thus
subsequently sepsis, the percentage distribution of the microbes depends on
the age
of the patient, the basic disorder, the primary infectious focus and the like
(Knaus,
W.A. et al. (1985), Crit. Care. Med. 13, 818-829). Leading microbes are both
Gram-
positive and Gram-negative bacteria, and fungi and yeasts (Llewelyn, M and
Cohen,
J. (2001), Int. Care Med. 27 S.10-S.32).
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Therapeutic methods and active ingredients which prevent the release of such
microbial products or bind and neutralize such products or influence
endogenous
functions and factors or lead to death of the microbes are suitable for the
treatment of
symptoms of the abovementioned diseases.
Substances and methods leading to a reduction in the number of microbes (e.g.
antibiotics), as well as substances having a circulation-influencing effect,
can be
utilized therapeutically (Forth, Henschler, Rummel; Allgemeine and spezielle
Pharmakologie and Toxikologie; Urban & Fischer Verlag (2001), Munich). The
standard therapy for the disease described above are antibiotics together with
substances or solutions which stabilize the circulation and influence
coagulation.
A further approach is to influence the pathological state by immunomodulatory
treatments in order to prevent or at least ameliorate an excessive response of
the body
to bacteria or bacterial products and thus avert organ failure (Zanotti, S. et
al. (2002),
Expert Opin. Investig. Drugs 11, 1061-1075).
In this connection, soluble mediators of the immune system which are supplied
as
therapy are also of particular interest. These mediators include inter alia
the so-called
defensins, molecules having antibacterial, antifungal or else antiviral
properties
(Kagan, B.L. et al. (1994), Toxicology 87, 131-149; Hancock R.E.W. and Scott
M.G.
,(2000), PNAS 16, 8856-8861). In addition, these molecules, which are small
peptides, have immune system-modulatory properties (Hancock R.E.W. and Scott
M.G. (2000), PNAS 16, 8856-8861). They bring about or prevent for example the
release of further mediators.
Even with optimal treatment according to the current therapeutic standard, the
mortality in cases of sepsis and septic shock is up to 50% of the patients
(Cohen, J.
(2002), Nature 420, 885-891). Additional therapeutic treatments are therefore
urgently necessary. However, in various approaches followed for the treatment
of
patients with severe bacteremias, sepsis and/or septic shock it has emerged,
even
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with newer therapies, that intervention in a disease process at one point is
unsuccessful or only partly successful. A combination of various therapeutic
approaches proves to be the most promising in the treatment of diseased
patients
(Anel, R.L. and Kumar, A. (2001) Expert. Opin. Investig. Drugs 10, 1471-1485).
A defensin from immune cells of the rhesus monkey (rhesus theta-defensin 1;
RTD-1) has been isolated and its properties, its broad antibacterial effect,
even on
non-growing bacteria, and its antifungal effect has been described in detail
(Tang et
al. (1999), Science 286, 498-502; Tran et al. (2002), J. Biol. Chem. 277, 3079-
3084)
and a patent application has been made therefor (WO 00168265). RTD-1 is
distinguished from other defensins or cationic peptides by some characteristic
and
defining properties. Firstly, RTD-1 is a small circular peptide and thus, in
contrast to
other defensins, stable to breakdown and degradation. RTD-1 shows no
dependence
of the effect on salts in the medium, and thus no loss of action in the
presence of
physiological concentrations of various salts such as, for example, NaCI, KCl
etc.
(Muhle et al. (2001), Biochemistry 40, 5777-5785; Tang et al. (1999), Science
286,
498-502), nor any loss of action in the presence of human serum (WO 00/68265).
In
addition, no hemolytic effect has been found.
In the invention described herein, surprisingly, a further effect of RTD-1 is
found in
states of bacteremia with subsequent sepsis and septic shock, and such
pathological
.states after exposure to bacterial products such as, for example, LPS. In
this
connection, an effect has been found on 4 different pathological states which
are
relevant for the syndrome of severe bacteremias, sepsis and septic shock. RTD-
1
intervenes in the pathological process by 1. showing an antimicrobial effect
on
various pathogens, 2. showing a neutralizing effect on bacterial products such
as LPS
or LTA (effect on products of Gram-positive as well as Gram-negative
bacteria),
which also permits prophylactic therapy, 3. having an immunomodulatory effect
in
the sense of mediator modulation, and 4. having a regulated anticoagulant
effect. 'The
combined effect on various parameters relevant to the disease results in an
unambiguously improved success of therapy together with simplified therapy. In
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addition, therapy with RTD-1 does not rule out current standard therapies
(e.g.
antibiotics, circulation-stabilizing substances) and allows combination
therapy.
Therapy with RTD-1 leads to increased survival of mice with severe bacteremia
after
administration of living bacteria, in particular both after infection with
Gram-positive
and Gram-negative bacteria. Moreover, surprisingly, no dependence is to be
found
with the minimum inhibitory concentration of RTD-1 on the bacterium used.
Therapy of mice with symptoms of septic shock after administration of LPS or
SEB
also shows a distinctly increased survival with RTD-1 therapy. Cytokine
analyses in
the serum or plasma of the animals show a regulation of the release of soluble
mediators. It is moreover possible to demonstrate a reduction of pro-
inflammatory
cytokines (such as, for example, TNF-a, IL-6, MIF) and an increase in
regulatory
factors (such as, for example, IF'N-y, IL-10). Comparable results are found in
human
whole blood, there being a reduction in the levels of pro-inflammatory
cytokines and
chemokines.
Under the influence of RTD-1 there is a dose-dependent increase in the
clotting time
of human plasma and human whole blood. RTD-1 thus shows an influence on the
coagulation parameters in human blood without an additional influence on
coagulation due to bacterial products such as, for example, LPS or LTA.
The present invention therefore relates to the use of theta-defensin from the
rhesus
monkey for producing a medicament for the treatment and/or prophylaxis of
bacterernias and/or sepsis.
1n this connection, the disease-causing agents may be Gram-positive bacteria,
Gram-
negative bacteria, bacterial products, viruses or yeast fungi.
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The invention further relates to the use of RTD-1 for producing a medicament
for
binding bacterial products such as LPS and/or LTA.
The invention further relates to the use of RTD-1 for producing a medicament
for the
treatment of pathological states characterized by changes in blood clotting.
The applications mentioned can moreover be combined with standard antibiotics
or
standard antimycotics.
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Examples
Example 1
Test for antibacterial effect in vitro
To determine the minimum inhibitory concentration, bacteria of various strains
are
exposed to graded concentrations of RTD-1, using a 1:2 dilution series.
Determination takes place according to the principles of the NCCLS (see
documentation: Methods for dilution antimicrobial susceptibility tests for
bacteria,
NCCLS document M7-A5, Vol. 20 No. 2).
Table 1: Minimum inhibitory concentrations of RTD-1, vancomycin and ampicillin
for various bacterial species determined by the NCCLS method. The table
indicates
the concentration of the respective compounds which showed unambiguous
inhibition of the growth of the bacteria.
Bacterium MIC [mg/L]
RTD-1 Vancom
cin Am
icillin
S. aureus MSSA 1 <0.125 0.5
S. aureus MRSA 8 1 >64
E. faecalis 2 2 2
-E. faecium 1 0.5 4
E. faecium VRE 0.5 >64 64
S. pneumoniae 64 0.25 0.125
E. coli 16 >64 8
P. aeruginosa 32 >64 >64
K. pneumoniae 8 >64 2
S. lyphimurium 8 >64 4
Table 1: Minimum inhibitory concentration of RTD-1, vancomycin and ampicillin
for various bacterial species
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Example 2
In vivo investigations in disease-relevant animal models
Bacterial suspensions are administered intraperitoneally (i.p.) to CFW-1 mice.
The
mice are purchased from Harlan. After 30 min, the animals are then treated
intravenously (i.v.) with RTD-1 in various doses. The survival of the animals
with
and without therapy reveals the success of therapy.
Table 2: Survival of mice after i.p. infection with S. aureus in the bacterema
model
and therapy with 0.1, 1 and 10 mg/kg RTD-1 i.v.. The mice are infected i.p.
with
1.68 x 107 colonies of S. aureus ATCC Smith and, after 30 min, are treated
with the
stated dose i.v.. The table indicates the % of survivors from n= 6 mice on day
5 after
infection.
Dose % Survival
No RTD-1 (control) 17
0.1 mg/kg 6'1
1 mg/kg 83
10 mg/kg 83
Table 2: Survival of S. aureus-infected mice with and without RTD-1 therapy
Table 3: Survival of mice after i.p. infection with S. pneumoniae in the
bacterema
model and therapy with 0.1, 1 and 10 mg/kg RTD-1 i.v.. The mice are infected
i.p.
with 3 _x 103 colonies of S. pneumoniae L3TV and, after 30 min, are treated
with the
stated dose i.v.. The table indicates the % of survivors from n= 6 mice on day
5 after
infection.
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Dose % Survival
No RTD-1 (control) 17
0.1 mg/kg 50
1 mg/kg 67
mglkg 33
Table 3: Survival of S. pneumoniae-infected mice with and without RTD-1
therapy
Table 4: Survival of mice after i.p. infection with E. coli in the bacterema
model and
5 therapy with 0.1, 1 and 10 mg/kg RTD-1 i.v.. The mice are infected i.p. with
1.68 x 107 colonies of E. coli Neumann and, after 30 min, are treated with the
stated
dose i.v.. The table indicates the % of survivors from n= 6 mice on day 5
after
infection.
Dose % Survival
No RTD-1 (control) 33
0.1 mg/kg 50
1 mg/kg 83
10 mg/kg 83
Table 4: Survival of E. coli-infected mice with and without RTD-1 therapy
Example 3
In addition, LPS is injected i.p. into mice and, at various times before and
after the
LPS administration, RTD-1 is administered i.v. in various dosage in order to
simulate
a septic shock. The survival of the animals is the measure of the success of
therapy in
the model of septic shock.
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Table 5: Survival of mice after i.p. administration of 20 mg/kg LPS from
S. typhimurium (Sigma) and therapy with 0.1, 1 and 10 mg/kg RTD-1 i.v. at
various
times before and after LPS administration. The table indicates the % of
survivors
from n= 5 mice on day 5 after LPS administration.
Dose %
Survival
-2h
-lh
+O.lh
+lh
+2h
No RTD-1 (control) 0
0.1 mglkg 100 100 100 100 80
1 mg/kg 100 100 100 100 100
mg/kg 80 80 100 80 80
Table 5: Survival of LPS-treated mice with and without RTD-1 therapy
Example 4
Investigations on the binding of bacterial products
The binding of RTD-1 to lipopolysaccharide (LPS) and lipoteichoic acid (LTA)
is
investigated in a binding mixture in vitro. This entails use of dansyl-
polymyxin B
(Moore et al. (1986), Antimicrob. Agents Chemotherap. 29, 496-500) which
fluoresces after binding to LPS or LTA. Competition with RTD-1 results in a
.decrease in the fluorescence and, resulting therefrom, a determination of the
relative
inhibition of the binding of dansyl-polymyxin B.
From the inhibition it is possible in turn to calculate a relevant affinity of
RTD-1 for
binding of bacterial products.
Table 6: LPS and LTA binding by RTD-1 and polymyxin B after incubation for
4 hours. The concentration at which the fluorescence of dansyl-polymyxin B is
reduced by 50% is indicated.
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Test substance 50% LPS binding at 50% LTA binding at
~M ~M
RTD 1 0.001 0.1
Polymyxin B 4.9 0.01
Table 6: LPS and LTA binding by RTD-1 and polymyxin B.
Example 5
The membrane permeability of bacteria under influence of RTD-1 is investigated
using a fluorescence method (Silvestro et al. (2000), Antimicrob. Agents
Chemotherap. 44, 602-607). This permits assessment of the potential of RTD-1
in
relation to damage to the cell membrane of bacteria and thus on the release of
bacterial products.
Table 7: Membrane potential change under the influence of RTD-1 and polymyxin
B
after exposure of S. aureus bacteria for 10 minutes. The concentration leading
to a
50% change in the fluorescence signal is shown.
Test substance 50% change in fluorescence
at
~M
RTD1 3.9
Polymyxin B >75
Table 7: Membrane potential change under the influence of RTD-1 and polymyxin
B.
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Example 6
In addition, the influence of RTD-1 on bacterial cell wall synthesis is
investigated.
This entails RTD-1 being added to a membrane fraction from E. coli (as example
of
Gram-negative bacteria) or from Bacillus megaterium (as example of Gram-
positive
bacteria) and the necessary substrates, and the inhibitory activity being
determined
(Chandrakala, B. et al. (2001) Antimicrob. Agents Chemother. 45, 768-775).
This
approach determines the incorporation of radioactive precursors in high
molecular
weight peptidoglycan via binding to wheat germ agglutinin. An influence of RTD-
1
on bacterial cell wall synthesis provides substantial information about an
effect on
the bacterium (e.g. lysis) and especially in connection with the release of
bacterial
products, which is a substantial precondition for assessing the therapeutic
potential of
RTD-1 in serious infections including septic shock.
Table 8: Inhibition of cell wall synthesis by RTD-1, ampicillin (Sigma), a
lactam
antibiotic, chloramphenicol (Sigma), a protein biosynthesis inhibitor and
vancomycin
(Sigma), a glycopeptide antibiotic. The inhibition of the binding to wheat
germ
agglutinin of radioactive precursors in high molecular weight peptide or
glycan is
shown. Only substances influencing cell wall biosynthesis but not inhibiting
protein
biosynthesis show inhibition in this approach.
Test substance Gram-negative Gram-positive
50% inhibition at 50% inhibition at ~.M
pM
Ampicillin 8.8 5.0
Chloramphenicol > 100 > 100
Vancomycin 4.4 0.85
RTD 1 7.2 5.7
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Table 8: Inhibition of cell wall synthesis in Gram-negative and Gram-positive
bacteria and bacterial lysates by RTD-l, ampicillin, vancomycin and
chloramphenicol.
Example 7
Investigation of the influence on the prothrombin time (PT)
This parameter is employed to determine disturbances in the extrinsic system
of
blood clotting. The prothrombin time is determined on citrated plasma after
addition
of calcium and tissue factor. For this purpose, blood is taken from healthy
people of
both genders in collecting tubes with citrate (Monovetten, Sarstedt,
Niunbrecht,
Germany) and the plasma is obtained after centrifugation. Samples of this
plasma are
incubated with various concentrations of the test compounds at 37°C for
10 minutes.
Then thromplastin (Recombiplastin, OrthoDiagnostic Systems, Neckargemiind,
Germany) is added in order to initiate the extrinsic pathway of blood
clotting. This
mixture is mixed and the clotting time is determined in an apparatus for
determining
coagulation (Coagulometer, Amelung, KC 4A micro).
Investigations of the influence on the partial thromboplastin time (aPTT)
,This parameter is employed to determine disturbances in the intrinsic system
of blood
clotting. The prothrombin time is determined on citrated plasma after addition
of an
activator and phospholipid. For this purpose, blood is taken from healthy
people of
both genders in collecting tubes with citrate (Monovetten, Sarstedt,
Niirnbrecht,
Germany) and the plasma is obtained after centrifugation. Samples of this
plasma are
incubated with various concentrations of the test compounds at 37°C for
10 minutes.
The activator kaolin and phospholipid (aPTT Reagent, Diagnostica Stago,
Asnieres,
France) is then added. Coagulation is started by adding 0.025 M calcium
chloride to
this mixture. This mixture is mixed and the clotting time is determined in an
apparatus for determining coagulation (Coagulometer, Amelung, KC 4A micro).
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Table 10: Influence on the coagulation parameters aPTT, PT, and the clotting
time of
human whole blood under the influence of RTD-1. The clotting time in seconds
after
addition of RTD-1 is shown.
Clotting time
RTD-1 concentration[sec] PT Whole blood
in ~g/ml aPTT
0 (control) 33.9 14.55 351.94
35.4 15.7 368.2
36.6 21.2 375.2
30 43.4 51.3 475.4
50 112.7 67.0 598.3
100 297 (stopped) 102.8 1000 (stopped)
Table 10: Influence on the coagulation parameters aPTT, PT, and the clotting
time of
human whole blood under the influence of RTD-1.
10 Example 8
In vivo cytolane investigations in disease-relevant animal models
At various times after LPS or SEB administration to mice, blood samples are
obtained by exsanguination of the animals. Plasma samples are drawn from these
blood samples and subjected to a cytokine analysis. The amount of cytokines in
the
plasma is determined quantitatively using the CBA methods (CBA system,
BectonDickinson, Heidelberg, Germany).
Table 11: Cytokine modulation after therapy with RTD-1 in the mouse model of
septic shock after administration of LPS. The maximum percentage change
compared
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with the untreated LPS control is indicated. Negative values indicate a
reduction, and
positive values an increase, in the amount of cytokines in the plasma.
Cytokine Change as %
of the untreated
LPS control
on intravenous
therapy with
RTD-1 0.1 mglkg
RTD-1 1 mg/kg
RTD-1 10 mg/kg
TNF-a. -47. 5 -41.1 -12.4
R,-6 -41.6 -5.8
IL-10 +157 +159 +241
gN_Y +53 +138 +69
Table 11: Cytokine modulation after therapy with RTD-1 in the mouse model of
septic shock after administration of LPS.
Table 12: Cytokine modulation after therapy with RTD-1 in the mouse model of
septic shock after. administration of SEB. The maximum percentage change
compared with the untreated SEB control is indicated. Negative values indicate
a
reduction, and positive values an increase, in the amount of cytokines in the
plasma.
Cytokine Change as %
of the untreated
SEB control
on intravenous
therapy with
RTD-1 0.1 mg/kg
RTD-1 1 mglkg
RTD-1 10 mglkg
TNF-a -3 2. 3 -29.7 -3 9.3
g-N-Y +8.2 +4.6 +28.6
Table 12: Cytokine modulation after therapy with RTD-1 in the mouse model of
septic shock after administration of SEB.
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Example 9
Ex vivo cytokine investigations in infected human blood
Blood is taken from healthy donors and infected in vitro with bacteria. Both
Gram-
positive pathogens (S. aureus) and Gram-negative pathogens (E. coli) are used
in this
case. After addition of the pathogens, samples of the infected and of the
infected and
treated blood are taken at defined times, and the plasma is obtained by
centrifugation.
The amount of cytokines in the plasma is determined quantitatively by the CBA
methods (CBA system, BectonDickinson, Heidelberg, Gemany). In addition, MIF is
analyzed by the ELISA technique (human MIF ELISA System, R&D Systems Inc.,
Minneapolis, USA).
Table 13: Cytokine modulation after therapy with RTD-1 in the infection model
with
human whole blood after administration of S. aureus. The maximum percentage
change in pro-inflammatory mediators compared with the untreated infection
control
is indicated. Negative values indicate a reduction, and positive values an
increase, in
the amount of cytokines in the blood culture.
Cytokine Change as %
of the untreated
infection
control on
therapy with
RTD-1 0.1 ug/ml
RTD-1 1 qg/ml
RTD-1 10 gg/ml
-
TNF-a -79 -85 -43
IL-113 -71 -73 -10
IL-6 -58 -73 -3
IL-8 -17 -25 -3
MIF -48 -44 -4
Table 13: Cytokine modulation after therapy with RTD-1 in the infection model
with
human whole blood after administration of S. aureus.
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Table 13: Cytokine modulation after therapy with RTD-1 in the infection model
with
human whole blood after administration of E. coli. The maximum percentage
change
compared with the untreated infection control is indicated. Negative values
indicate a
reduction, and positive values and increase, in the amount of cytokines in the
blood
culture.
Cytokine Change as % of
the untreated
infection control
on
therapy with
RTD-1 10 ~.g/ml
RTD-1 100 pg/ml
TNF-a -8 -4
IL-6 -43 -20
MIF -77 -41
Table 14: Cytokine modulation after therapy with RTD-1 in the infection model
with
human whole blood after administration of E. coli
Formulations
RTD-1 can be converted in a known manner into the usual formulations such as
tablets, coated tablets, pills, granules, aerosols, syrups, emulsions,
suspensions and
-solutions, using inerk, nontoxic, pharmaceutically suitable carriers or
solvents. In
these cases, the therapeutically effective compound is to be present in each
case in a
concentration of from 0.5 to 90% by weight of the complete mixture, i.e. in
amounts
which are sufficient to achieve the indicated dosage range.
The formulations are produced for example by extending the active ingredients
with
solvents and/or carriers, where appropriate using emulsifiers and/or
dispersants, it
being possible for example when water is used as diluent where appropriate to
use
organic solvents as auxiliary solvents.
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Administration takes place in a conventional way, preferably intravenously,
transdermally, orally or parenterally, in particular orally or intravenously.
However, it
can also take place by inhalation through the mouth or nose, for example with
the aid
of a spray, or topically through the skin.
It has generally proved to be advantageous to administer amounts of about
0.001 to
mg/kg, on oral administration preferably about 0.005 to 3 mg/kg, of bodyweight
to achieve effective results.
It may be necessary where appropriate to deviate from the stated amounts, in
particular as a function of the bodyweight or the nature of the administration
route,
the individual response to the medicament, the nature of its formulation and
the time
or interval over which administration takes place. Thus, it may be sufficient
in some
cases to make do with less than the aforementioned minimal amount, whereas in
other cases the stated upper limit must be exceeded. If larger amounts are
administered, it may be advisable to divide these into a plurality of single
doses over
the day.
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Literature list:
Chandrakala, B. et al. (2001 ) Antimicrob. Agents Chemother. 45, 768-775
Forth, Henschel, Rummel (2001), Allgemeine and spezielle Pharmakologie and
Toxikologie; Urban&Fischer Verlag, Munich
Moore et al. (1986), Antimicrob. Agents Chemotherap. 29, 496-500
Muhle et al. (2001), Biochemistry 40 5777-5785
Silvestro et al. (2000), Antimicrob. Agents Chemotherap. 44, 602-607
Tang et al. (1999), Science 286, 498-502
Tran et al. (2002), J. Biol. Chem. 277. 3079-3084
List of abbreviations:
ATCC American Type Culture Collection
LPS Lipopolysaccharide
LTA Lipoteichoic acid
MIC Minimum inhibitory concentration
NCCLS National Committee for Clinical Laboratory Standards
RTD Rhesus theta defensin
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SEQUENCE LISTING
<110> Bayer AG
<120> Treatment of serious infections and septic shock
<130> Le A 36 086
<160> 1
<170> PatentIn version 3.1
<210> 1
<211> 18
<212> PRT
<213> rhesus monkey
<400> 1
Gly Phe Cys Arg Cys Leu Cys Arg Arg Gly Val Cys Arg Cys Ile Cys
1 5 10 15
Thr Arg
