U.S. patent application number 11/378798 was filed with the patent office on 2006-12-14 for methods and compositions for the reduction of neutrophil influx and for the treatment of bronchpulmonary dysplasia, respiratory distress syndrome, chronic lung disease, pulmonary fibrosis, asthma and chronic obstructive pulmonary disease.
Invention is credited to Aprile L. Pilon.
Application Number | 20060281681 11/378798 |
Document ID | / |
Family ID | 38522965 |
Filed Date | 2006-12-14 |
United States Patent
Application |
20060281681 |
Kind Code |
A1 |
Pilon; Aprile L. |
December 14, 2006 |
Methods and compositions for the reduction of neutrophil influx and
for the treatment of bronchpulmonary dysplasia, respiratory
distress syndrome, chronic lung disease, pulmonary fibrosis, asthma
and chronic obstructive pulmonary disease
Abstract
The present invention relates generally to the use of
recombinant human CC10 (rhCC10), also known as recombinant human
uteroglobin, for use as a therapeutic in the treatment of
Respiratory Distress Syndrome (RDS), Bronchopulmonary dysplasia
(BPD), chronic lung disease and/or pulmonary fibrosis, Asthma and
Chronic Obstructive Pulmonary Disease (COPD). More particularly,
the invention provides methods, including broadly the critical
dosage ranges of rhCC10, which may be administered to safely and
effectively treat the aforementioned conditions. The invention
further provides a composition useful in administering rhCC10 to
humans.
Inventors: |
Pilon; Aprile L.;
(Germantown, MD) |
Correspondence
Address: |
KRAMER LEVIN NAFTALIS & FRANKEL LLP;INTELLECTUAL PROPERTY DEPARTMENT
1177 AVENUE OF THE AMERICAS
NEW YORK
NY
10036
US
|
Family ID: |
38522965 |
Appl. No.: |
11/378798 |
Filed: |
March 16, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11189229 |
Jul 25, 2005 |
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11378798 |
Mar 16, 2006 |
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09835784 |
Apr 13, 2001 |
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11189229 |
Jul 25, 2005 |
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09549926 |
Apr 14, 2000 |
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09835784 |
Apr 13, 2001 |
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09120264 |
Jul 21, 1998 |
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09549926 |
Apr 14, 2000 |
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09087210 |
May 28, 1998 |
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09120264 |
Jul 21, 1998 |
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08864357 |
May 28, 1997 |
6255281 |
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09087210 |
May 28, 1998 |
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Current U.S.
Class: |
435/69.1 ;
514/1.5; 514/1.7 |
Current CPC
Class: |
A61P 11/00 20180101;
A61K 38/17 20130101; A61K 38/1709 20130101 |
Class at
Publication: |
514/012 |
International
Class: |
A61K 38/17 20060101
A61K038/17 |
Claims
1. A method for treatment of a patient having respiratory distress
syndrome, bronchopulmonary dysplasia, chronic lung disease,
pulmonary fibrosis, asthma or chronic obstructive pulmonary disease
and in need of treatment, comprising: administering a
treatment-effective amount of rhCC10 to said patient.
2. A method for treatment of a patient having respiratory distress
syndrome, bronchopulmonary dysplasia, chronic lung disease,
pulmonary fibrosis, asthma or chronic obstructive pulmonary disease
and in need of treatment, comprising the steps of: (a) monitoring
said patient's serum concentration of CC10 and (b) administering a
treatment-effective dose of rhCC10 which causes the patient to
reach a peak serum concentration of CC10 within about 6 hours.
3. A method for treatment of a premature infant having respiratory
distress syndrome, bronchopulmonary dysplasia, asthma or chronic
obstructive pulmonary disease and in need of treatment, comprising
administering rhCC10 intratracheally to a premature infant at a
dose of between about 0.15 mg/kg to about 5 mg/kg of patient body
mass.
4. A method for treatment of a premature infant having respiratory
distress syndrome, bronchopulmonary dysplasia, asthma or chronic
obstructive pulmonary disease and in need of treatment, comprising
administering rhCC10 intratracheally to a premature infant at a
dose of between about 1.5 mg/kg to about 5 mg/kg of patient body
mass.
5. A method for treatment of a premature infant having respiratory
distress syndrome, bronchopulmonary dysplasia, asthma or chronic
obstructive pulmonary disease and in need of treatment, comprising
administering rhCC10 intratracheally to a premature infant at a
dose of between about 1.5 mg/kg to about 5 mg/kg of patient body
mass before, during, or after surfactant therapy.
6. A safe and well-tolerated method of treating at least one of
respiratory distress syndrome, bronchopulmonary dysplasia, chronic
lung disease, pulmonary fibrosis, asthma or chronic obstructive
pulmonary disease in a patient comprising: administering rhCC10 to
the patient.
7. The method of claim 6 wherein between about 15 ng to about 10 mg
rhCC10 per kg of patient body mass is administered.
8. The method of claim 6 wherein between about 0.15 mg to about 5
mg rhCC10 per kg of patient body mass is administered.
9. The method of claim 6 wherein between about 1.5 mg and about 5
mg rhCC10 per kg of patient body mass is administered.
10. The method of claim 6 wherein IL-8 levels are less than about
120 pg/ml of serum after administration.
11. The method of claim 6 wherein the patient has a serum
concentration of CC10 of about 100 ng/ml of serum to about 2800
ng/ml of serum or a urine concentration of CC10 of about 100 ng/ml
to about 10,000 ng/ml during treatment.
12. The method of claim 6 wherein the patient's peak serum
concentration of CC10 is about 1280 ng/ml of serum to about 2800
ng/ml of serum after administration.
13. The method of claim 6 wherein the patient reaches a peak serum
concentration of CC10 within about 6 hours after initial
dosage.
14. The method of claim 6 wherein the patient reaches a peak
tracheal aspirate fluid concentration of CC10 between about 12 and
about 48 hours after initial dosage.
15. The method of claim 6 wherein the administration of rhCC10 is
repeated about once every 48 hours.
16. A safe and well-tolerated method of reducing total cell counts
in the tracheal fluid of a patient comprising: administering rhCC10
to the patient.
17. The method of claim 16 wherein neutrophil levels in the lungs
of a patient are reduced.
18. The method of claim 16 wherein rhCC10 between about 15 ng to
about 10 mg per kg of patient body mass is administered.
19. The method of claim 16 wherein rhCC10 between about 0.15 mg to
about 5 mg per kg of patient body mass is administered.
20. The method of claim 16 wherein rhCC10 between about 1.5 mg and
about 5 mg per kg of patient body mass is administered.
21. The method of claim 16 wherein the patient's serum IL-8 level
is less than 120 pg/ml after administration.
22. The method of claim 16 wherein the patient has a serum
concentration of CC10 of about 100 ng/ml of serum to 2800 ng/ml of
serum or a urine concentration of CC10 of about 100 ng/ml to about
10,000 ng/ml during treatment.
23. The method of claim 16 wherein the patient's peak serum
concentration of CC10 is about 1280 ng/ml of serum to about 2800
ng/ml of serum after administration.
24. The method of claim 16 wherein a peak serum concentration of
CC10 is reached within about 6 hours after initial dosage.
25. The method of claim 16 wherein the patient reaches a peak TAF
concentration of CC10 between about 12 and about 48 hours after
initial dosage.
26. The method of claim 16 wherein the administration of rhCC10 is
repeated about once every 48 hours.
27. A safe and well-tolerated method of reducing total protein
concentration in the tracheal fluid of a patient comprising:
administering rhCC10 to the patient.
28. The method of claim 27 wherein neutrophil levels in the lungs
of a patient are reduced.
29. The method of claim 27 wherein between about 15 ng to about 10
mg rhCC10 per kg of patient body mass is administered.
30. The method of claim 27 wherein between about 0.15 mg to about 5
mg rhCC10 per kg of patient body mass is administered.
31. The method of claim 27 wherein between about 1.5 mg and about 5
mg rhCC10 per kg of patient body mass is administered.
32. The method of claim 27 wherein the patient's serum IL-8 level
is less than 120 pg/ml after administration.
33. The method of claim 27 wherein the patient has a serum
concentration of CC10 of about 100 ng/ml of serum to 2800 ng/ml of
serum or a urine concentration of CC10 of about 100 ng/ml to about
10,000 ng/ml during treatment.
34. The method of claim 27 wherein the patient's peak serum
concentration of CC10 is about 1280 ng/ml of serum to about 2800
ng/ml of serum after administration.
35. The method of claim 27 wherein a peak serum concentration of
CC10 is reached within about 6 hours after initial dosage.
36. The method of claim 27 wherein the patient reaches a peak
tracheal aspirate fluid concentration of CC10 between about 12 and
about 48 hours after initial dosage.
37. The method of claim 27 wherein the administration of rhCC10 is
repeated about once every 48 hours.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation-in-part of pending
application Ser. No. 11/189,229, filed Jul. 25, 2005, which is a
continuation-in-part of application Ser. No. 09/835,784, filed Apr.
13, 2001, now abandoned, which is a continuation-in-part of
application Ser. No. 09/549,926, filed Apr. 14, 2000, now
abandoned, which is a continuation-in-part of application Ser. No.
09/120,264, filed Jul. 21, 1998, now abandoned, which is a
continuation-in-part of application Ser. No. 09/087,210, filed May
28, 1998, now abandoned, which is a continuation-in-part of
application Ser. No. 08/864,357, filed May 28, 1997, U.S. Pat. No.
6,255,281. Each of the aforementioned applications and patent are
incorporated herein by reference in their entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to methods of reducing the
influx of neutrophils into the lungs of a human. More specifically
the present invention relates to methods of treating respiratory
distress syndrome (RDS) bronchopulmonary dysplasia (BPD), chronic
lung disease, pulmonary fibrosis, asthma and chronic obstructive
pulmonary disease (COPD) in humans and compositions useful for the
same. Yet more specifically, the present invention relates to
methods of treating the above using recombinant human CC10 and
compositions useful for the same.
BACKGROUND
[0003] The influx of neutrophils into the lung is known to be a
cause of the destruction of functional lung tissue and the harmful
symptoms of RDS, BPD, chronic lung disease, pulmonary fibrosis,
asthma and COPD. Neutrophil influx is the migration of neutrophils
from the blood into tissue in response to any type of irritation or
injury to the tissue. In RDS, BPD, chronic lung disease and/or
pulmonary fibrosis, asthma and COPD total cell influx and
neutrophil influx results in damage to and destruction of pulmonary
tissue, ultimately causing functional lung tissue to be replaced
with non-functioning fibrotic tissue. Thus, neutrophil influx is
ultimately responsible for causing pulmonary fibrosis to the lungs,
which lead to damaged lung tissue and possibly death.
[0004] When circulating neutrophils are activated by chemical and
cytokine signals released by damaged or irritated tissue, for
example, by IL-8 from the lungs, they adhere to the walls of blood
vessels in the damaged tissue, following the chemical and cytokine
signal to the source, and migrate through the vascular endothelia
and into the damaged lung. Neutrophils release many powerful
enzymes, such as myeloperoxidase (MPO), an enzyme that chemically
damages and modifies all proteins in its local vicinity, usually
inactivating them. MPO damages local lung tissue and proteins, as
well as those of the infectious agents. Neutrophils also release
powerful proteases, such as elastase, that indiscriminately degrade
host and pathogen proteins alike. Thus, activated neutrophils that
migrate into the lungs in response to some irritation of the
respiratory tract release non-specific destructive enzymes that
damage the host's respiratory tissues, as well as any infectious
agents present. These, and other, powerful non-specific destructive
mediators released by neutrophils damage all cell types and destroy
lung tissue, including breaking down the bronchiolar and alveolar
structure, resulting in decreased lung function and respiratory
distress. Local vascular structure is also damaged, resulting in
increased vascular permeability and leakage of serum proteins into
the lung and tracheal fluid, further impairing function.
[0005] This non-specific neutrophil response results in greater
damage to respiratory tissue than the original irritant or
infectious agent. Patients that develop respiratory symptoms or
respiratory distress as a result of an exposure to an irritant such
as an allergen, air-borne particulate matter, chemicals, or
infectious agents often experience the worst symptoms after the
irritant or infectious agent is cleared from the respiratory tract.
Neutrophils are substantially responsible for this
over-reaction.
[0006] Neutrophil influx to the lungs is measured in patients by
counting the number of neutral-staining white cells per unit volume
in tracheal fluids (referred to as tracheal aspirate fluid or TAF).
Tracheal fluids are continuous with bronchial fluids, alveolar
fluids, and nasal and sinus fluids. Tracheal fluid composition is
representative of pulmonary fluids in the lower respiratory tract,
as well as the upper respiratory tract (nasal and sinus fluid).
[0007] Cytokines, like IL-6 and IL-8, are released by local
epithelial cells, endothelial cells, and fibroblasts. The levels of
cytokines, for example IL-6 and IL-8, are measured in lung fluids
such as TAF and in plasma or serum. Cytokines are basic regulators
of all neutrophil functions. Under normal conditions, neutrophils
move along microvascular walls via low affinity interaction of
selectins with specific endothelial carbohydrate ligands. However,
during the inflammatory response, chemotactic factors and
proinflammatory cytokines signal the recruitment of neutrophils
(neutrophil influx) to sites of infection and/or injury.
Neutrophils then penetrate the endothelial layer and migrate
through connective tissue to sites of injury, for example the
lungs, where they accumulate and adhere to extracellular matrix
components such as fibronectin and/or collagen.
[0008] RDS affects 10% of all premature infants and only rarely
affects those born at full-term. RDS also affects adults. The
disease is caused by a lack of lung surfactant, a chemical that
normally appears in mature lungs, or by tissue damage to the lungs
from being on a mechanical ventilator and oxygen for a significant
amount of time. Surfactant keeps the air sacs from collapsing and
allows them to inflate with air more easily. In respiratory
distress syndrome, the air sacs collapse and prevent the child from
breathing properly. In infants, symptoms usually appear shortly
after birth and become progressively more severe. If symptoms of
RDS persist, the condition is considered BPD if a baby is dependent
on artificially supplied oxygen at 36 weeks' postconceptional age
(PCA--also known as post-conceptual age). In a child or adult, if
symptoms of RDS persist, the condition is considered chronic lung
disease and/or pulmonary fibrosis if the patient is dependent on
artificially supplied oxygen following a respiratory distress
episode.
[0009] BPD affects 20-60% of all premature, very low birth weight
infants. BPD and RDS are associated with substantial morbidity and
mortality as well as extremely high health care costs. Although the
widespread use of intratracheally administered exogenous surfactant
and antenatal steroid therapy has reduced the overall severity of
BPD, the prevalence of this condition has increased with improved
survival of very low birth weight infants. BPD is a multi-factorial
disease process that is the end result of an immature, surfactant
deficient lung that has been exposed to hyperoxia, mechanical
ventilation and infection. Furthermore, it is well documented that
increased concentrations of cytokines and cells present in the
tracheal aspirate fluid of premature infants within the first few
days of life are associated with the subsequent development of RDS
and BPD. Still further it is known that higher levels of
fibronectin are present in the tracheal fluid and lungs of patients
suffering from RDS and BPD and thus it causes and contributes to
respiratory distress. Thus, treating and preventing RDS and BPD by
providing improved lung function during the first few days of life
of a premature infant is critical to the long term survivability of
the infant.
[0010] Asthma is a chronic lung condition characterized by
difficulty in breathing. Symptoms include: wheezing, coughing
shortness of breath and chest tightness. People with asthma have
extra sensitive or hyperresponsive airways. The airways react by
narrowing or obstructing when they become irritated. This makes it
difficult for the air to move in and out. This narrowing or
obstruction causes the symptoms of asthma. The narrowing or
obstruction of the airways is caused by: airway inflammation
(meaning that the airways in the lungs become red, swollen and
narrow) or bronchoconstriction (meaning that the muscles that
encircle the airways tighten or go into spasm)
[0011] COPD is a lung disease in which the lungs are damaged,
making it hard to breathe. In COPD, the airways are partly
obstructed, making it difficult to get air in and out of the lungs.
Most cases of chronic obstructive pulmonary disease (COPD) develop
after repeatedly breathing in fumes and other things that irritate
and damage the lungs and airways, for example by smoking. The lungs
and airways are highly sensitive to irritants. They cause the
airways to become inflamed and narrowed, and they destroy the
elastic fibers that allow the lung to stretch and then return to
its resting shape. This makes breathing air in and out of the lungs
more difficult. COPD may also be caused by a gene-related disorder
called alpha 1 antitrypsin deficiency. Alpha 1 antitrypsin is a
protein that inactivates destructive proteins. People with
antitrypsin deficiency have low levels of alpha 1 antitrypsin; the
resulting imbalance of proteins leads to the destruction of the
lungs and COPD.
[0012] Symptoms common to RDS, BPD, chronic lung disease, pulmonary
fibrosis, asthma and COPD include respiratory insufficiency (i.e.
lungs unable to adequately oxygenate the blood and remove carbon
dioxide), increased airway resistance, inflammation and fibrosis of
the lungs. Each of these symptoms are substantially caused by
excessive levels of neutrophils, IL-6, IL-8, and total cells in the
tracheal fluid. Excess total protein in the tracheal aspirate fluid
("TAF") or bronchoalveolar lavage fluid ("BAL") is also associated
with lung inflammation and fibrosis in RDS, BPD, chronic lung
disease, pulmonary fibrosis, asthma and COPD.
[0013] Glucocorticoids, also known as corticosteroids, are powerful
anti-inflammatory agents that are known to improved lung function,
to reduce the incidence of BPD in premature infants, and to improve
the symptoms of RDS, BPD, chronic lung disease and/or pulmonary
fibrosis, asthma and COPD. However, they are not completely safe to
use. There are dangerous, often life-threatening side effects
associated with the use of glucocorticoids in infants, children and
adults. In infants, corticosteroids are avoided in clinical
practice because they cause growth retardation, disproportionate
growth inhibition of the central nervous system and head, and
severe neurological impairment. In children, normal growth is
stunted, resulting in small stature, due to treatment with
corticosteroids. And in adults, cardiovascular complications,
including hypertension and stroke, are major side effects of
corticosteroids. In all patients, corticosteroids lower the
patient's immune function and leave them susceptible to infection
of all types (bacterial, viral, fungal, etc.), sometimes resulting
in a lethal infection. Thus, safety is a major consideration in the
choice of anti-inflammatory agent used to treat, prevent or cure
RDS, BPD, chronic lung disease and/or pulmonary fibrosis, asthma
and COPD and their related respiratory symptoms, reduce the
severity of asthma or allergy, and prevent the progression of
existing chronic lung disease such as COPD or the development of
chronic lung disease such as BPD. It is a significant challenge to
find an anti-inflammatory agent powerful enough to alleviate
respiratory symptoms and which is safe to use.
[0014] Human CC10 (hereinafter CC10), also known as uteroglobin, is
a small homodimeric secretory protein produced by mucosal
epithelial cells. In humans, Clara cells, a type of mucosal
epithelial cell located in the airways, are the main site of CC10
production. CC10 also circulates in the blood and is excreted in
urine. CC10 is known to have anti-inflammatory properties. CC10
inhibits secretory PLA.sub.2, an enzyme that degrades surfactant
and facilitates eicosanoid biosynthesis. Eicosanoids are a family
of lipophilic compounds including prostaglandins, leukotrienes,
thromboxanes, and other arachidonic acid metabolites.
[0015] Further information concerning rhCC10, its structure and
methods of use is found in U.S. Pat. No. 6,255,281, its
continuation-in-part, U.S. patent application Ser. No. 09/087,210,
and in the following U.S. Patent Application Publication Nos.: US
2002-0160948, US 2002-0160948, US 2003-0008816, US 2003-0109429, US
2003-0207795, US 2002-0173460, US 2002-0169108, US 2005-0261180, US
2004-0047857, and US 2006-0025348, all of which are incorporated by
reference in their entirety.
[0016] Very low concentrations of CC10 have been found in the TAF
or BAL of patients suffering from RDS, Asthma and COPD and BPD. For
example, very low concentrations of CC10 have been found in the
tracheal aspirate fluid (TAF) of ventilated premature infants
suffering from BPD relative to normal levels. These infants are not
yet able to produce enough natural CC10 on their own, and develop
severe lung inflammation. Normally, the appearance of CC10 in the
amniotic fluid dates from 16 weeks of gestation and increases as a
function of gestational as well as postnatal age. CC10
concentrations in tracheal fluid, measured by determining the
amount of CC10 protein in the tracheal aspirate fluid, of premature
infants born at 28-32 weeks of gestation have been found to be 2-4
orders of magnitude less than those found in the tracheal sputum
(a.k.a. tracheal fluid) of healthy adults. CC10 concentrations
correlate in a negative fashion with the concentration of inspired
oxygen required by preterm infants with BPD. That is, infants with
lower CC10 in TAF require greater amounts of supplemental oxygen.
In fact, not only are CC10 concentrations lower in tracheal fluid
from infants who either died or developed BPD, but the limited
amount of available CC10 was oxidized and demonstrated less
immunoreactivity relative to controls.
[0017] Recombinant CC10 (recombinant human CC10) has not been
previously used to treat patients, including preterm infants for a
number of reasons. First, rhCC10 of sufficient purity has not been
previously available. Nor was it known whether rhCC10 caused
specific toxicity or triggered an immune response to endogenous
CC10 when administered. Furthermore, CC10 is known to inhibit
platelet aggregation, thus negatively impacting the ability of the
blood to clot. CC10 is also known to suppress the immune system,
which could lead to adverse patient consequences, render recipients
more susceptible to infection, and prohibit its use in humans,
including premature infants. It was not known what dosage or dosage
range would avoid deleterious immunogenicity, specific toxicity,
and inhibition of platelet aggregation and suppression of the
immune system.
[0018] Furthermore, it was not known whether rhCC10 would cause
significantly lower total protein concentrations in the tracheal
fluids of patients or what dosage to administer to achieve
significantly lower total protein concentrations in the tracheal
fluid of patients, a necessary outcome in treating BPD.
[0019] Additionally, it was not known whether rhCC10 would cause
significantly lower total cell, neutrophil, IL-6 or IL-8 levels in
patients or which dosage to administer to achieve significantly
lower neutrophil, IL-6 or IL-8 levels in patients, also a necessary
outcome in treating RDS, BDP, chronic lung disease and/or pulmonary
fibrosis, Asthma, and COPD.
[0020] As shown below, the prior technological difficulties in
using CC10 to provide a safe, well-tolerated and effective
treatment for RDS, BDP, chronic lung disease, pulmonary fibrosis,
asthma, and COPD have been overcome.
OBJECTS OF THE INVENTION
[0021] The foregoing provides a non-exclusive list of the
objectives achieved by the present invention:
[0022] It is a primary object of the invention to treat, cure or
prevent RDS, BDP, chronic lung disease and/or pulmonary fibrosis,
asthma, and COPD in humans.
[0023] It is a further object of the invention to treat, cure or
prevent RDS, BDP, chronic lung disease and/or pulmonary fibrosis,
asthma, and COPD in humans by reducing total cell counts,
neutrophil counts, total protein concentration IL-6 levels and/or
IL-8 levels in the serum or tracheal fluid, and therefore the
lungs, of patients.
[0024] It is a further object of the invention to provide a safe,
well-tolerated and effective dosage range which accomplishes the
above objectives and does not significantly inhibit platelet
aggregation, suppress the immune response or increase the frequency
or severity of adverse events.
[0025] It is yet another object of the invention to provide a safe,
well-tolerated and effective dosage which provides a substantially
effective range of CC10 levels in patient serum, tracheal fluid and
urine.
SUMMARY OF THE INVENTION
[0026] These and other objects, features and advantages are
achieved by administering rhCC10 in a dosage range given at
appropriate intervals, or in one dose to treat, cure or prevent
RDS, BDP, chronic lung disease, pulmonary fibrosis, asthma, and
COPD.
[0027] These and other objects, features and advantages are also
achieved by administering rhCC10 in a dosage range given at
appropriate intervals or in one dose where a patient shows one or
more of the following: IL-6 levels below 200 pg/ml of tracheal
aspirate fluid, IL-8 levels below 100 pg/ml in serum, total
neutrophil cell counts below 20 cells.times.10.sup.4/ml of tracheal
aspirate fluid, total cell counts below 50 cells.times.10.sup.4/ml
of tracheal aspirate fluid, and total protein concentration below
400 .mu.g/ml of tracheal aspirate fluid, such dosing being
continued until the RDS, BDP, chronic lung disease and/or pulmonary
fibrosis, asthma, or COPD has been treated, cured, or
prevented.
[0028] These and other objects, features and advantages are also
achieved by administering rhCC10 such that it does not inhibit
platelet aggregation, suppress the immune response or increase the
frequency or severity of adverse events.
[0029] In certain aspects of the invention, rhCC10 is administered
intratracheally in a dose between about 1.5 and about 5.0 mg/kg of
patient body mass or in multiple doses which taken together achieve
this dosage range to treat, cure or prevent RDS, BDP, chronic lung
disease and/or pulmonary fibrosis, asthma, or COPD. In another
aspect, an rhCC10 dose or doses adding up to between about 1.5 and
about 5.0 mg/kg of patient body mass may be repeated at appropriate
intervals to treat, cure or prevent RDS, BDP, chronic lung disease
and/or pulmonary fibrosis, asthma, or COPD. In yet other aspects of
the invention, rhCC10 is administered intratracheally in accordance
with the above aspects but in a dose or doses adding up to between
about 15 nanograms/kg of patient body mass and about 10 mg/kg of
patient body mass or in a dose adding up to between about 0.15
mg/kg and about 5 mg/kg of patient body mass. Whether administered
intratracheally or otherwise, rhCC10 may be given alone, in
conjunction with, before or after surfactant therapy.
BRIEF DESCRIPTIONS OF THE DRAWINGS
[0030] FIG. 1 is a bar graph of the CC10 concentration in the
tracheal aspirate fluid of patients over time that were given
placebo, rhCC10 at 1.5 mg/kg of body mass or rhCC10 at 5 mg/kg of
body mass.
[0031] FIG. 2 is a bar graph of the CC10 concentration in the serum
of patients over time that were given placebo, rhCC10 at 1.5 mg/kg
of body mass or rhCC10 at 5 mg/kg of body mass.
[0032] FIG. 3 is a bar graph of the CC10 concentration in the urine
of patients over time who were given placebo, rhCC10 at 1.5 mg/kg
of body mass or rhCC10 at 5 mg/kg of body mass.
[0033] FIG. 4 is a bar graph of total cell counts in the tracheal
aspirate fluid of patients over time who were given placebo, or
rhCC10 at 5 mg/kg of body mass.
[0034] FIG. 5 is a bar graph of total neutrophil counts in the
tracheal aspirate fluid of patients over time who were given
placebo or rhCC10 at 5 mg/kg of body mass.
[0035] FIG. 6 is a bar graph of total protein concentration in the
tracheal aspirate fluid of patients over time who were given
placebo, rhCC10 at 1.5 mg/kg of body mass or rhCC10 at 5 mg/kg of
body mass.
[0036] FIG. 7 is a bar graph of IL-6 levels in the tracheal
aspirate fluid of patients over time who were given placebo, rhCC10
at 1.5 mg/kg of body mass or rhCC10 at 5 mg/kg of body mass.
[0037] FIG. 8 is a bar graph of IL-8 levels in the serum of
patients at 48 hours after administration who given placebo, rhCC10
at 1.5 mg/kg of body mass or rhCC10 at 5 mg/kg of body mass.
DETAILED DESCRIPTION
[0038] The present invention relates to the critical dosages and
timing of administration of rhCC10 to treat, cure or prevent RDS,
BDP, chronic lung disease and/or pulmonary fibrosis, asthma, and
COPD in humans. rhCC10 is preferably obtained by the processes
described in U.S. Patent Application Publication Nos. US
2003-0109429 and US 2003-0207795, both of which are incorporated by
reference in their entirety, or via any other process which yields
pharmaceutical grade rhCC10. The rhCC10 of the embodiments of the
present invention may also be administered by the intratraceal,
endotracheal, dialysate, ophthalmic, intravenous, systemic, or oral
routes. Furthermore the rhCC10 of the embodiments of the present
invention may be administered with, without, before or after
surfactant therapy.
[0039] Preferably, in treating or preventing RDS or BPD rhCC10 is
administered during the first day of an infant patient's life. More
preferably, rhCC10 is administered as soon as medically possible
during the first day of an infant patient's life, for example, and
without limitation, within about 30 minutes of intubation and
receipt of surfactant.
[0040] Premature infants are typically intubated for the purposes
of administering oxygen and inflating their lungs, which would
collapse without intubation. Intubation can then serve as a direct
route for the intratracheal administration (also known as
endotracheal administration) of medicines, such as surfactants, to
the lungs. Thus the preferred route of administration for rhCC10 is
also intratracheal administration, however alternate routes of
administration are also possible such as inhalation, inhalation of
pegylated rhCC10, injection into the muscle tissues, intravenous
injection, intranasal administration, oral administration and
administration by suppository.
[0041] rhCC10 may also be administered to treat, cure or prevent
chronic lung disease and/or pulmonary fibrosis, asthma, and COPD.
Based on the results described herein, rhCC10 will have therapeutic
benefit to patients suffering from chronic lung disease and/or
pulmonary fibrosis, asthma, and COPD. More specifically, and as
shown below, rhCC10, when dosed at the amounts described below,
lowers neutrophil counts, IL-6 levels and IL-8 levels in humans,
and thus will provide an effective treatment for RDS, Asthma and
COPD.
[0042] One method of measuring the therapeutic effect of rhCC10 on
a patient is to measure conditions present in the tracheal aspirate
fluid, which is indicative of conditions present in the tracheal
and bronchoalveolar fluid of the patient's lungs. Such conditions
may be one or more of the following: IL-6 levels, IL-8 levels,
total neutrophil counts, total cell counts and total protein
concentration.
[0043] A method of measuring the concentration of CC10 in a
patient, and thus determining whether treatment has established
therapeutically effective levels of CC10 in the patient, may
include one or more of following: measuring CC10 concentration in
TAF, serum or urine.
[0044] With reference to the following embodiments, rhCC10 may be
administered to achieve certain desired effects, establishing that
therapeutically effective levels of CC10 have been achieved in the
patient, while at the same time avoiding other deleterious effects.
For example, rhCC10 may be administered to achieve concentrations
of CC10 in the tracheal aspirate fluid which exceed the deficient
production of endogenous CC10 by the premature infant. rhCC10 may
also be administered to achieve early peak serum concentrations of
rhCC10 in patients, for example within about 6 hours after
administration. As a further example, when administered in
accordance with the described methods, rhCC10 administration may
also achieve a peak concentration in urine at about 12 hours for
example. As yet another example, rhCC10 may be administered so as
to achieve significantly lower cell counts, neutrophil cell counts,
protein concentrations, IL-6 levels and IL-8 levels in the
patient's tracheal aspirate fluid and in the patient's lungs.
[0045] Furthermore, rhCC10 may be administered such that it does
not significantly reduce the patient's endogenous CC10 production,
inhibit platelet aggregation or cause an adverse immunologic
reaction.
[0046] To effectuate the desired outcomes which are further
described below, reference is made to methods of administration
described in the following embodiments:
[0047] In one embodiment, a dose or multiple doses of rhCC10
equaling a dose ranging from about 1.5 to about 5 mg/kg of body
mass may be administered. In another embodiment a dose or multiple
doses of rhCC10 equaling a dose ranging from about 15 nanograms/kg
of body mass to about 10 mg/kg of body mass is administered. In
still another embodiment a dose or multiple doses of rhCC10
equaling a dose ranging from about 0.15 mg/kg and about 5 mg/kg of
patient body mass is administered.
[0048] In yet another embodiment the above doses of rhCC10 may be
administered intratracheally to the patient. In yet another
embodiment, the above doses of rhCC10 may be administered to the
patient by aerosol. In a further embodiment rhCC10, in accordance
with the methods described above, may be administered prior to,
during or after surfactant therapy. In still another embodiment,
rhCC10, in accordance with the methods described above, may be
administered to treat RDS, BPD, asthma, chronic lung disease,
pulmonary fibrosis or COPD in a patient.
[0049] The doses of rhCC10 described above may be administered
daily, more than once daily, every other day or in a tapered
fashion depending upon the severity of disease being treated, the
patient's overall health, and whether an acute or chronic condition
is being treated. For example, the more severe the disease
condition, a higher the amount of rhCC10 would be required to
effectively treat the disease. For maintenance therapy of chronic
disease, for example, to prevent an exacerbation of chronic Asthma,
RDS, COPD, BDP or other pulmonary condition, lower doses would be
required. It is understood that a physician would be able to
monitor and adjust doses as needed based on the patient's symptoms
and responses to therapy and within the parameters and dose ranges
described in the embodiments of the present invention.
[0050] The following detailed examples are illustrations of
embodiments. It should be clear that these are not intended to
limit the scope of the present invention.
EXAMPLE 1
Administration of rhCC10 to Premature Infants
[0051] Patients were enrolled in a placebo-controlled, blinded,
dose ranging study at four hospital sites.
[0052] rhCC10 was produced in E. coli bacteria and purified by a
proprietary process (Claragen, Inc., College Park, Md.), described
in U.S. Application Publication Nos. US 2003-0109429 and US
2003-0207795, both of which are incorporated by reference in their
entirety. The protein for the study was provided as a >98% pure
solution of recombinant human CC 10 homodimer. The biological
activity of each batch was compared using a proprietary secretory
PLA.sub.2 inhibition assay, described in U.S. Application
Publication Nos. US 2002-0169108 which is incorporated herein by
reference.
[0053] Newborn infants who met the following criteria were
enrolled: 1) age .ltoreq.24 h; 2) birth weight between 700 and
1,300 g; 3) gestational age .gtoreq.24 wk; 4) diagnosis of RDS
based on clinical and radiographic criteria; 5) requirement for
intubation and mechanical ventilation; 6) receipt of surfactant,
100 mg/kg (Survanta, Ross Laboratory). Patients could be given
subsequent doses of surfactant if clinically indicated following
rhCC10 administration. Table 1 depicts the composition of the study
groups (cohorts): TABLE-US-00001 TABLE 1 Study Population
Gestational Birth Race Age Weight Sex (White/Black/ Any Maternal
(weeks) (grams) (Male/Total) Hispanic/Asian) Steroids Placebo 26.5
.+-. 1.6 943 .+-. 137 5/7 (71%) 3, 2, 1, 1 7/7 1.5 mg/kg 27.6 .+-.
1.2 981 .+-. 159 3/7 (43%) 6, 2, 0, 0 6/7 5.0 mg/kg 26.5 .+-. 1.2
878 .+-. 205 3/6 (50%) 4, 2, 1, 0 6/6
[0054] rhCC10, was formulated in a volume of 2 ml/kg of sterile,
unbuffered saline. Patients were enrolled in two cohorts, each
comparing study drug to placebo. The first cohort consisted of 12
patients, randomized so that one-third received placebo and the
other two-thirds received rhCC10 at 1.5 mg/kg of body mass of the
study drug. After the first cohort of patients was enrolled and the
safety data reviewed by the DSMC, a second cohort of 12 patients
was enrolled. They were also randomized so that one-third received
placebo and the other two-thirds received rhCC10 at 5.0 mg/kg of
body mass.
[0055] Each patient then received a single dose of the study drug
(or placebo) as soon as possible after surfactant replacement
therapy, but not longer than 4 h after surfactant. Study drug or
placebo was administered intratracheally (IT) in two equal aliquots
via a pre-measured feeding tube placed into the distal third of the
endotracheal tube, with the patient in the right and then left
lateral decubitus position and 30.degree. of Trendelenburg.
[0056] As described in greater detail in Examples 2-4,
pharmacokinetic analyses were conducted on samples of TAF, serum,
and urine samples and analyses of cells counts and protein levels
were performed on samples of TAF.
[0057] TAF was obtained by instilling 1 ml of saline into the
endotracheal tube and suctioning the fluid into a Leuken's trap.
The catheter was then washed with an additional 1 ml saline. In
some cases the first tracheal aspirate was obtained prior to
surfactant administration (baseline). Subsequent TAF collections
were obtained at 12, 24, 48 and 72 hours post-administration. TAF
was only collected if infants continued to require intubation and
mechanical ventilation. The TAF was centrifuged at 300.times.g for
10 minutes to pellet the cells. The supernatant was removed and
frozen at -70.degree. C.
[0058] The pharmacokinetic analysis was conducted on samples of
TAF, serum, and urine samples using a competitive ELISA for human
CC10 developed by the sponsor. The assay utilizes a single
anti-human CC10 polyclonal antibody as a capture reagent. CC10 in
the sample competes with a synthetic CC10-HRP (horseradish
peroxidase) conjugate for antibody binding sites in the plate
wells. Thus, the signal decreases with increasing CC10
concentration in the sample. Samples were run in duplicate and a
standard curve was run for each set of assays using rhCC10
calibrators. The limit of detection is 5 ng/ml and the results were
reproducible with coefficients of variation typically under 20%.
The assay does not appear to distinguish between native and
recombinant CC10, thus total CC10 levels were measured.
Immunogenicity of the study drug was assessed by titration of
anti-C10 antibodies in serum obtained at 28 days post
administration.
[0059] Referring to Examples 5-8 below, analyses for pulmonary
inflammatory markers were performed as follows: The TAF cell pellet
was resuspended and cell counts performed using a hemocytometer.
Differential cell counts in TAF were determined by
cytocentrifugation and differential staining. Total protein in TAF
was measured using the Pierce BCA technique, and a panel of
cytokines (Multiplex cytokine analysis, Luminetics Corp.) was
measured in TAF from all three experimental groups at 0, 24 and 48
hours post-administration. IL-6 and IL-8 cytokines were measured in
TAF from patients in all three groups at times 0, 1 and 2 days
(with a minimum of three and maximum of seven samples/group).
[0060] Concentrations of CC10 and analysis of inflammatory markers
over time were examined by using mixed model analysis of variance
to test the interaction of time and dose. Non-parametric testing
was performed when unequal variance was detected. Sample
characteristics, the incidence of complications and clinical
outcomes were analyzed by Fisher's Exact Test for categorical
variables or one way analysis of variance for continuous
variables.
EXAMPLE 2
TAF Concentrations of CC10 in Patients Treated with rhCC10
[0061] With reference to FIG. 1 it has been found that during the
first 48 hours of life, after an initial dose of rhCC10,
significantly increased overall CC10 concentration occurred in
patients receiving rhCC10 in dosages comprising either 1.5 mg/kg of
body mass or 5 mg/kg of body mass versus placebo. Thus,
administration of rhCC10 during the first 24 hours of life has a
significant positive impact on CC10 levels in patients during the
first two days of life. Furthermore, administration of rhCC10 will
increase overall CC10 concentrations in patients.
[0062] Reference is now made to Table 2, as well as to FIG. 1, the
contents of which are further described in this example.
TABLE-US-00002 TABLE 2 Average TAF CC10 Concentrations CC10 CC10
CC10 CC10 Conc. in Conc. in Conc. in Conc. in TAF** TAF TAF TAF 12
Hours 24 Hours 48 Hours 72 Hours Placebo 476 ng/ml 753 ng/ml 916
ng/ml 3435 ng/ml 1.5 mg/kg* 2336 ng/ml 1639 ng/ml 2492 ng/ml 1522
ng/ml rhCC10 5 mg/kg* 2400 ng/ml 1994 ng/ml 1432 ng/ml 784 ng/ml
rhCC10 *dosage units are mg of rhCC10 per kg of patient body mass
**CC10 concentration in TAF are in units of ng of CC10 per ml of
TAF
[0063] CC10 concentrations in patients were measured at 12, 24, 48
and 72 hours post-administration. An average concentration for each
patient group receiving a particular dose of rhCC10 (1.5 or 5
mg/kg) or placebo was determined as follows. CC10 concentrations in
TAF were observed for time points where there were at least three
patient samples per group (FIG. 1). This allowed for analysis of
TAF samples for all groups following administration of placebo
(0.9% sterile saline) or rhCC10 through day 3 of life. For safety
and logistical reasons, samples were not obtained if surfactant had
recently been administered or if the infant had been extubated. At
12 hours of life, CC10 concentrations in TAF from infants treated
with the study drug was significantly higher than the placebo
group, but there was little difference between the two groups who
received rhCC10. Over the first 3 days of life, CC10 concentrations
in TAF from infants receiving placebo generally increased, whereas
CC10 levels in treated infants tended to remain constant (1.5 mg/kg
of body mass) or decrease (5 mg/kg of body mass). However, CC10
levels from infants receiving placebo did not exceed the CC10
levels of those infants receiving rhCC10 in either 1.5 mg/kg of
body mass or 5 mg/kg of body mass dosages during the first 48 hours
of life. Those infants receiving 1.5 mg/kg of body mass rhCC10 had
the highest levels of CC10 at 48 hours.
EXAMPLE 3
Serum Concentrations of CC10 in Patients Treated with rhCC10
[0064] Furthermore, with reference to FIG. 2, in one embodiment,
rhCC10 may be administered intratracheally such that peak serum
levels of CC10 are achieved within 6 hours of administration. Peak
serum levels occur within 6 hours, irrespective of the dose of
rhCC10 administered. Based on the results described below and in
FIG. 2, peak serum levels will occur within about 6 hours after
administration across all dosage ranges.
[0065] Reference is now made to Table 3, as well as to FIG. 2, the
contents of which are further described in this example.
TABLE-US-00003 TABLE 3 Average Serum CC10 Concentrations CC10 CC10
CC10 CC10 CC10 CC10 Conc. in Conc. in Conc. in Conc. in Conc. in
Conc. in serum** serum** serum** serum** serum** serum**
Elimination 0 Hours 6 Hours 12 Hours 24 Hours 36 Hours 48 Hours
Half-life Placebo 42 ng/ml 43 ng/ml 46 ng/ml 40 ng/ml 37 ng/ml 38
ng/ml Not applicable 1.5 mg/kg* 76 ng/ml 1289 ng/ml 782 ng/ml 354
ng/ml 196 ng/ml 101 ng/ml 11.6 hours rhCC10 5 mg/kg* 22 ng/ml 2794
ng/ml 1476 ng/ml 798 ng/ml 290 ng/ml 143 ng/ml 9.9 hours rhCC10
*dosage units are mg of rhCC10 per kg of patient body mass **CC10
concentration in serum are in units of ng of CC10 per ml of
serum
[0066] In determining average peak serum levels, blood (0.3 ml) was
obtained for the measurement of serum concentration of CC10 before
drug administration (0 hours) and at 6, 12, 24, 36, and 48 hours
after administration of rhCC10. An average concentration for each
patient group receiving a particular dose of rhCC10 or placebo was
determined.
[0067] Serum concentrations of CC10 were similar in all 3 groups
before treatment (FIG. 2). Infants who received rhCC10 had
substantially higher serum concentrations than infants receiving
placebo and this varied in a dose dependent manner. Average peak
serum levels after administration of rhCC10 may range from about
1290 ng/ml of serum to about 2800 ng/ml serum when rhCC10 is given
in a single dose of between 1.5 mg/kg of body mass and 5 mg/kg of
body mass. As shown in Table 3, the elimination half-life of a
rhCC10 dosage of 1.5 mg/kg of body mass was about 11.6 hours,
whereas the elimination half-life of a rhCC10 dose of 5 mg/kg of
body mass was about 9.9 hours. CC10 concentrations in the serum of
treated infants were comparable to placebo levels within 48 hours
of administration.
EXAMPLE 4
Urine Concentrations of CC10 in Patients Treated with rhCC10
[0068] Referring to FIG. 3, in one embodiment, rhCC10 may also be
administered intratracheally at the above-mentioned dosages such
that peak CC10 levels in urine occur 12 hours after administration.
For example, CC10 concentrations in the urine of treated infants
increase in a largely dose-dependent manner, but are comparable to
placebo levels within 48 h of administration.
[0069] Reference is now made to Table 4, as well as to FIG. 3, the
contents of which are further described in this example.
TABLE-US-00004 TABLE 4 Average Urine CC10 Concentrations CC10 CC10
CC10 Conc. in CC10 Conc. in Conc. in Conc. in urine** urine**
urine** urine** 12 Hours 24 Hours 36 Hours 48 Hours Placebo 197
ng/ml 183 ng/ml 180 ng/ml 203 ng/ml 1.5 mg/kg* 3312 ng/ml 1226
ng/ml 1513 ng/ml 593 ng/ml rhCC10 5 mg/kg* 9239 ng/ml 2613 ng/ml
763 ng/ml 583 ng/ml rhCC10 *dosage units are mg of rhCC10 per kg of
patient body mass **CC10 concentration in serum are in units of ng
of CC10 per ml of urine
[0070] Urine samples were obtained at 12, 24, 36 and 48 hours after
administration of rhCC10. Each urine sample consisted of the total
volume voided over the previous 12 hours.
[0071] In urine, CC10 concentrations in treated infants increased
in a largely dose-dependent manner, but were comparable to placebo
levels within 48 h of administration (FIG. 3).
EXAMPLE 5
Total Cell Counts in TAF of Patients Treated with rhCC10
[0072] As shown in FIG. 4 and in Table 5, total cell counts were
performed on TAF fluids and are shown in FIG. 5. Average total cell
counts were obtained by measuring and averaging total cell counts
in TAF within the placebo and 5 mg/kg rhCC10 study groups. Study
groups were sampled at 0.5, 1, 2, and 3 days post-administration.
TABLE-US-00005 TABLE 5 Total Cell Counts in TAF Total cell Total
cell Total cell Total cell count** count** count** count** 0.5 days
1 day 2 days 3 days Placebo 1.0 36 61 56 5 mg/kg* 9.8 11 24 14
rhCC10 *dosage units are mg of rhCC10 per kg of patient body mass
**total cell counts are in units of Cells .times. 10.sup.4 per ml
of TAF
[0073] Total cell counts were significantly lower in the 5 mg/kg
group on days 1-3 compared to the placebo group. Total cells counts
were at least twice as low during days 1-3 of life after rhCC10 at
5 mg/kg of body mass was administered versus placebo.
EXAMPLE 6
Total Neutrophil Counts in TAF in Patients Treated with rhCC10
[0074] Total neutrophil counts were performed on TAF fluids in
order to gauge rhCC10's effect on inflammation in the lungs and are
shown in FIG. 5. Inflammation of the lungs is caused by an excess
of neutrophil cells which are a cause of RDS, BDP, chronic lung
disease, pulmonary fibrosis, asthma, and COPD. Average neutrophil
counts were obtained by measuring and averaging neutrophil counts
in TAF within each study group (placebo and 5 mg/kg rhCC10). Study
groups were sampled at 0.5, 1, 2, and 3 days post-administration.
TABLE-US-00006 TABLE 6 Total Neutrophil Counts in TAF Total cell
Total cell Total cell Total cell count** count** count** count**
0.5 days 1 day 2 days 3 days Placebo 0.1 13.7 31.2 12 5 mg/kg*
rhCC10 2.4 3.4 10.2 4.7 *dosage units are mg of rhCC10 per kg of
patient body mass **total cell counts are in units of Cells .times.
10.sup.4 per ml of TAF
[0075] Neutrophil counts were significantly lower in the 5 mg/kg
group relative to the placebo group. For example, on day two the
placebo group's neutrophil levels were over 30 cells.times.10(4)
/ml of TAF versus about 10 cells.times.10(4) /ml of TAF for the
group receiving rhCC10 at 5 mg/kg of body mass. Therefore,
excessive neutrophil cell amounts were minimized in the lungs.
EXAMPLE 7
Total Protein Concentration in TAF of Patients Treated with
rhCC10
[0076] Referring now to FIG. 6 and to Table 7, total protein levels
were measured in the TAF of both treatment groups (rhCC10 at 1.5
mg/kg and 5 mg/kg of body mass) in order to gauge rhCC10's effect
on protein leak and pulmonary edema. Both protein leak and
pulmonary edema are conditions damaging to the lungs and
symptomatic of RDS, BDP, chronic lung disease, pulmonary fibrosis,
asthma, and COPD. Average total protein concentrations were
obtained by measuring and averaging total protein concentrations in
TAF within each study group (placebo, 1.5 mg/kg rhCC10 and 5 mg/kg
rhCC10). Study groups were sampled at 0.5, 1, 2, and 3 days
post-administration. TABLE-US-00007 TABLE 7 Total Protein
Concentrations in TAF Total Total Total Total protein protein
protein protein conc.** conc.** conc.** conc.** 0.5 days 1 day 2
days 3 days Placebo 307 430 536 964 1.5 mg/kg* 565 527 685 264
rhCC10 5 mg/kg* 289 334 189 167 rhCC10 *dosage units are mg of
rhCC10 per kg of patient body mass **total cell counts are in units
of .mu.g of protein per ml of TAF
[0077] Total protein was significantly lower in TAF from both
treatment groups compared to placebo. For example, by day three
post-administration, total protein in the TAF of the placebo group
was nearly 1000 .mu.g/ml of TAF whereas total protein in the
treatment groups on day three did not exceed 350 .mu.g/ml of TAF.
Thus protein leak and pulmonary edema had been minimized in the
treatment groups.
EXAMPLE 8
Total IL-6 Levels in TAF CC10 in Patients Treated with rhCC10
[0078] IL-6 cytokine was measured in TAF from patients in all three
groups at times 0, 1 and 2 days post-administration (with a minimum
of three and maximum of seven samples/group).
[0079] Referring now to Table 8 and FIG. 7, IL-6 concentrations
were effectively reduced by the study drug (rhCC10 in saline) in
both groups, but increased over time in the placebo group.
TABLE-US-00008 TABLE 8 Pharmacokinetic Results: IL-6 Levels in TAF
IL-6 IL-6 IL-6 IL-6 level level** level** level** 0 days 0.5 days 1
day 2 days Placebo n/a 150 291 449 1.5 mg/kg* 572 187 269 99 rhCC10
5 mg/kg* 446 64 103 113 rhCC10 *dosage units are mg of rhCC10 per
kg of patient body mass **total cell counts are in units of pg of
IL-6 per ml of TAF
[0080] For example, those patients receiving rhCC10 at 5 mg/kg of
body mass had IL-6 levels below 200 pg/ml of TAF over the first two
days following administration. Those patients receiving rhCC10 at
1.5 mg/kg of body mass had IL-6 levels below 300 pg/ml of TAF over
the first two days post-administration and had an IL-6 level below
100 pg/ml by day two. However, those patients on placebo had
steadily increasing IL-6 levels, exceeding 400 pg/ml by day 2. Thus
rhCC10, when administered according to the present teachings,
reverses the upward trend of IL-6 levels in patient lungs, thus
preventing neutrophil influx to the lung.
EXAMPLE 9
Total IL-8 Levels in TAF and Serum in Patients Treated with
rhCC10
[0081] Referring now to FIG. 8 and Table 9, IL-8 cytokine was
measured in TAF from patients in all three groups at 48 hours
post-administration. IL-8 is a potent chemoattractant for
neutrophils and other circulating inflammatory cells, that is
released by local epithelial cells, resident immune cells, and
fibroblasts in response to injury or irritation. TABLE-US-00009
TABLE 9 Pharmacological Results: Total IL-8 Levels in Serum IL-8 in
Serum** 48 hours Placebo 118 pg/ml 1.5 mg/kg* rhCC10 89 pg/ml 5
mg/kg* rhCC10 72 pg/ml *dosage units are mg of rhCC10 per kg of
patient body mass **total IL-8 levels are in units of pg of IL-8
per ml of serum
[0082] IL-8 levels were lower in rhCC10-treated patients than in
patients receiving placebo. Administration of rhCC10, as depicted
above, reduced the levels of IL-8 released from the lungs and into
the systemic circulation. Thus, this data shows that rhCC10 is
effective at treating or preventing RDS, BDP, chronic lung disease
and/or pulmonary fibrosis, asthma, and COPD by lowering IL-8 levels
and thus treating the causative agent of these diseases.
EXAMPLE 10
Outcomes
[0083] Table 10 depicts comparative outcomes of patients who
received rhCC10 versus patients who received placebo. Patients were
monitored at six months of corrected age (the developmental
timepoint at which they would have been six months old had they
been born at the normal 40 weeks of gestation.) Table 11 depicts
further comparative outcomes of patients who received rhCC10 versus
patients who received placebo. TABLE-US-00010 TABLE 10 Outcomes at
6 Month Corrected Age Outcome Placebo 1.5 mg/kg 5 mg/kg # with
repeat 4/6 4/6 0/5 Respiratory symptoms (cough, wheezing) # with
doctor visits 4/6 2/6 3/5 for Respiratory symptoms # hospitalized
for 3/6 0/6 0/5 breathing problems
[0084] Referring to Table 10 above, patients who received rhCC10
therapy had reduced incidences of respiratory symptoms, e.g.
coughing and wheezing. Coughing and wheezing are symptoms common
RDS, BDP, chronic lung disease and/or pulmonary fibrosis, asthma,
and COPD. After receiving rhCC10, patients had fewer doctor visits
due to respiratory symptoms, and no patients were hospitalized for
breathing problems in comparison to 50% of infants in the placebo
group who were hospitalized for their respiratory symptoms. This
data shows that rhCC10 significantly reduces the severity of
respiratory symptoms, preventing the need for rehospitalization.
TABLE-US-00011 TABLE 11 Observations During Initial Hospitalization
Placebo 1.5 mg/kg 5 mg/kg Doses of Surfactant 1.9 +/- 10.7 1.4 +/-
0.7 1.5 +/- 0.5 Days on Ventilator 12.1 .+-. 8.6 8.2 .+-. 7.8 24
.+-. 13.1 Days on Ventilator 33 .+-. 12.7 18.7 .+-. 13.2* 44.3 .+-.
18.1 and NCPAP Days on O.sub.2 56.6 .+-. 13.1 49 .+-. 11.2 55 .+-.
18.1 O.sub.2 at 28 d 7/7 7/7 5/6 O.sub.2 at 36 wk PCA 2/7 1/7 3/6
Hospitalized at 36 wk 5/7 2/7 4/6 PCA (71.4%) (28.6%) (66.7%) PDA
5/7 3/8 2/7 Sepsis 0/0 1/8 1/7 IVH 2/7 0/8 1/7 PVL 1/7 0/8 0/7
NCPAP--nasal continuous positive airway pressure, NEC--necrotizing
enterocolitis; PCA--post-conceptual age, PDA--patent ductus
arteriosus, IVH--intraventricular hemorrhage, PVL--periventricular
leukomalacia
[0085] Referring to Table 11 above, the therapeutic effect of
rhCC10 on short term respiratory distress is reflected in the
decreased requirement for additional doses of exogenous surfactant
therapy. The length of hospital stays was tabulated for each study
group. Only 28.6% of patients in the low dose group were still
hospitalized after 36 weeks compared to 71.4% of patients in the
placebo group and 66.7% in the high dose groups.
[0086] Patients in the low dose rhCC10 group (1.5 mg/kg of body
mass) were on the ventilator and NCPAP for significantly fewer days
than the placebo patients. When total days of mechanical
ventilation were evaluated, there was also a trend towards a
reduction in the need for ventilatory support in the low dose group
(the 1.5 mg/kg body mass group).
[0087] These results show that rhCC10 therapy, when administered in
accordance with the present teachings, decreased the severity of
RDS compared to placebo and reduced or eliminated the incidence of
respiratory problems severe enough to warrant medical attention or
rehospitalization.
[0088] Furthermore, this data shows that the safety profile of
rhCC10 is superior to other anti-inflammatory agents such as
corticosteroids. The safety and tolerability of the study drug were
assessed through 36 weeks PCA (Post conceptual age, also known as
PMA--post menstrual age) by comparing the incidence of adverse
events in the treatment and placebo groups and to the historical
incidence of the adverse events at each institution. No deaths were
attributable to administration of rhCC10.
[0089] In addition, a preliminary assessment of the efficacy of IT
rhCC10 in decreasing the incidence of BPD was made on the basis of
the following data: duration of mechanical ventilation, oxygen
requirement at 28 days with an abnormal chest radiograph, oxygen
requirement at 36 weeks PCA or date of discharge.
[0090] Growth parameters were assessed at birth, 28 days of age and
36 weeks PCA. Blood chemistries and liver function tests were
evaluated at the onset of the study and on days 7 and 28
post-administration. Complete blood counts and urinalysis were
performed on enrollment, 24, 48 and 72 h, 7 and 28d
post-administration. Cranial ultrasounds were performed upon
randomization and were repeated at 7 and 28d of life.
EXAMPLE 12
Incidence of PVL and IVH Adverse Events
[0091] There were no instances of PVL (peri-ventricular
leukomalacia) in the rhCC10-treated infants. However, there was one
infant in the placebo group who developed PVL. PVL occurs when
leukocytes (primarily neutrophils) infiltrate the brain and cause a
severe inflammatory response. PVL, if not lethal, typically results
in severe neurological impairment in the infant. Smaller and
younger infants are predisposed to PVL. Even though the infants in
the high dose group (5.0 mg/kg patient body mass) were smaller and
younger than in the placebo group,there was no PVL in the high dose
group. rhCC10 thus protected these disadvantaged infants from
PVL.
[0092] Likewise, the incidence of IVH (Intraventricular
hemorrhage), which occurs when a blood vessel in the brain bursts
in response to aggressive ventilation and high oxygen levels, was
lower in the rhCC10 treated groups than in the placebo group.
RhCC10 appears to have protected these disadvantaged infants in the
high dose group from IVH, refuting the scientific papers that
taught that rhCC10 inhibits platelet aggregation and would promote
hemorrhaging in vivo.
[0093] PDA (patent ductus arteriosis), a defect or incomplete
closure in the walls of the heart, was significantly decreased in
the rhCC10-treated groups compared to placebo. PDA is a
life-threatening problem that must be corrected surgically, if it
is not resolved in the first several months of life. rhCC10 reduced
the incidence of PDA, possibly by decreasing the stress on the
heart.
[0094] There were no significant differences in values obtained for
blood chemistries, complete blood counts or results of urinalysis
among groups at any of the time points evaluated.
[0095] There were no significant differences in the incidence of
non-respiratory adverse events in the treatment and placebo groups
(Table 2). Three cases of NEC occurred at one center in
rhCC10-treated infants. However, other premature infants not
enrolled in the study at that center also developed NEC at the same
time. Growth parameters were similar among the groups.
EXAMPLE 13
Immunological Safety
[0096] It will be further appreciated that the safety of rhCC10 can
be measured by conducting an analysis of the potential
immunogenicity of the administered rhCC10 using plasma samples.
Plasma samples collected on day 28 of life were tested for the
presence of anti-CC10 antibodies. No evidence of antibody formation
was present in any of the 28 day plasma samples from any of the
groups.
EXAMPLE 14
Other Outcomes
[0097] With reference to Table 2, there were no significant
differences in the incidence of non-respiratory adverse events in
the treatment and placebo groups. Three cases of NEC occurred at
one center in rhCC10-treated infants. However, other premature
infants not enrolled in the study at that center also developed NEC
at the same time. Growth parameters at 36 weeks CGA were similar
among the groups. Similarly, there were no significant differences
in values obtained for blood chemistries, complete blood counts or
results of urinalysis among groups at any of the time points
evaluated. Thus, rhCC10 administration, in contrast to
corticosteroids, did not appear to cause any significant safety
issues in premature infants.
[0098] In addition, IT rhCC10 did not elicit an immunogenic
response from treated infants. The only adverse event that was
increased in treated infants compared to placebo controls was NEC
(p=NS), however, it was not possible to attribute the NEC to the
administration of rhCC10 for two reasons. First, excess CC10 was
cleared from all infants by 48h post-administration and the 3 cases
of confirmed NEC occurred 3-6 wk post-administration. Second, all
cases of confirmed NEC occurred at the same center. In addition,
there were other cases of NEC in infants not enrolled in the rhCC10
study in this center occurring in the same timeframe, suggesting an
outbreak pattern. These data indicate it is highly unlikely that
rhCC10 administration was related to the development of NEC.
[0099] Referring now to, for example, Examples 10-13, it has been
shown that rhCC10 is safe and well-tolerated because, upon
administration to a patient, it does not elicit any immediate or
delayed local or systemic reactions in the patient, it is not
associated with any unusual adverse events, or increased severity
or frequency of typical adverse events for the treated patient
population, as described above. Furthermore, rhCC10 is safe and
well-tolerated because it does not elicit any immunologic response
from the patient to either the rhCC10 or to endogenous CC10, does
not predispose the patient to bleeding or hemorrhage, or
specifically increase the platelet aggregation time in the patient,
and does not compromise the patient's immune function and
predispose the patient to infection.
EXAMPLE 15
[0100] A patient, who may be an adult, child or a premature infant
presents with RDS, BDP, chronic lung disease (in the case of a
child or an adult), pulmonary fibrosis (in the case of a child or
an adult), asthma, or COPD. A dose of rhCC10 from 1.5 mg/kg to 5
mg/kg of patient body mass is given to the patient. The patient
will then demonstrate a resolution of the symptoms of the
aforementioned condition.
EXAMPLE 16
[0101] A patient, who may be an adult, child or a premature infant
presents with RDS, BDP, chronic lung disease (in the case of a
child or an adult), pulmonary fibrosis (in the case of a child or
an adult), asthma, or COPD. A dose of rhCC10 from 0.15 ng/kg to 5
mg/kg of patient body mass is given to the patient. The patient
will then be relieved of the symptoms of the aforementioned
condition.
EXAMPLE 17
[0102] A patient, who may be an adult, child or a premature infant
presents with RDS, BDP, chronic lung disease (in the case of a
child or an adult), pulmonary fibrosis (in the case of a child or
an adult), asthma, or COPD. A dose of rhCC10 from 0.15 mg/kg to 5
mg/kg of patient body mass is given to the patient. The patient
will then be relieved of the symptoms of the aforementioned
condition.
[0103] Based on the foregoing, the critical ranges for rhCC10
dosages effective to safely treat, cure and prevent RDS, BDP,
chronic lung disease and/or pulmonary fibrosis, asthma, and COPD
have been found. Accordingly, the present invention provides a safe
and well-tolerated rhCC10 based therapy effective at treating the
symptoms of RDS, BDP, chronic lung disease and/or pulmonary
fibrosis, asthma, and COPD thus increasing the long term
survivability of both premature infants, child and adult patients
suffering from these conditions, while not causing any dangerous
side effects.
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