Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Peptide containing doai
Reexamination Certificate
1996-07-29
2001-06-12
Feisee, Lila (Department: 1646)
Drug, bio-affecting and body treating compositions
Designated organic active ingredient containing
Peptide containing doai
C514S014800, C514S015800, C514S016700, C514S017400, C435S875000, C530S350000, C530S402000, C530S300000
Reexamination Certificate
active
06245735
ABSTRACT:
BACKGROUND OF THE INVENTION
Cystic fibrosis (CF) is a disease that arises due to mutations in the gene that codes for cystic fibrosis transmembrane conductance regulator (CFTR), which is a membrane protein involved in chloride ion secretion [1]. Although most cystic fibrosis patients develop chronic progressive disease of the respiratory system, the disease can cause damage to many other organs and tissues. For instance, pancreatic dysfunction , hepatobiliary and genitourinary diseases are all common manifestations of the cystic fibrosis disorder. The diverse array of symptoms and disorders caused by cystic fibrosis have made treatment of the disorder a difficult task. Many treatment modes have focused on improving the clinical symptoms of the particular organ affected in the patient, such as antibiotic treatments, improved nutritional care, and physiotherapy. Additionally, therapies have been developed which attempt to counteract the biochemical basis of the genetic disease, such as gene therapy with CFTR genes. None of these treatment methods, however, has been entirely successful in the treatment of cystic fibrosis.
The most serious consequence of cystic fibrosis (CF) is
Pseudomonas aeruginosa
lung infection, which by itself accounts for almost 90% of the morbidity and mortality in CF [3]. By age 12, 60-90% of CF patients are infected with
P. aeruginosa,
and most die before age 30 [3]. Pathogens such as
S. aureus
and nontypable
H. influenzae
are also commonly isolated from the respiratory tract of CF patients, but only
P. aeruginosa
infection has been associated with the progressive decline in pulmonary function in these patients [4-6].
Progressive loss of pulmonary function over many years due to chronic infection with mucoid
P. aeruginosa
is the hallmark of CF, and yet the connection between lung infection and defects in chloride ion conductance have remained elusive. Smith et al. [2] recently reported defective bacterial killing by fluid obtained from airway epithelial cell cultures of CF patients. Smith et al. reported that this phenomenon was due to the inhibition of an unidentified antimicrobial factor resulting from increased levels of sodium chloride in the airway epithelial fluid.
Many of the severe cases of CF are associated with CFTR mutations leading to greatly reduced to no cell-surface expression of CFTR. The most prevalent of the CFTR mutations is the deletion of phenylalanine 508. Mutant CFTR genes having a deleted phenylalanine 508 are referred to as &Dgr;F508. &Dgr;F508 accounts for approximately 70% of the cystic fibrosis alleles. The &Dgr;F508 mutation has been associated with elevated sweat chloride levels and severe physiological effects such as chronic pulmonary disease in many patients.
Pier et al. has proposed that ingestion and clearance of
P. aeruginosa
by epithelial cells could be one mechanism by which the epithelial cells protect the lungs against infection [7]. The study reported that ingestion and clearance of
P. aeruginosa
was compromised in a cell line derived from a patient with the &Dgr;F508 CFTR mutation and was specific for
P. aeruginosa
among the respiratory pathogens evaluated [7]. Expression of wild-type CFTR by transection, or induction of membrane expression of mutant &Dgr;F508 CFTR by growth of cells at 26° C., increased
P. aeruginosa
ingestion. Inhibition of ingestion of
P. aeruginosa
by cells in neonatal mouse lungs increased the total bacterial load in the lungs [7]. These studies showed that CFTR modulated this epithelial cell process but did not specifically indicate how CFTR was involved in the process.
SUMMARY OF THE INVENTION
The invention involves the discovery that
P. aeruginosa
binds to the cystic fibrosis transmembrane conductance regulator (CFTR) (SEQ.ID.NO.1) and, in particular, that the core portion of the lipopolysaccharide of
P. aeruginosa
binds the CFTR. The invention also involves the discovery that contacting cells expressing the CFTR with the core portion of the lipopolysaccharide of
P. aeruginosa
results in upregulation of the CFTR. Upregulation of the CFTR in epithelial mucosa further was discovered to result in better clearance of
P. aeruginosa
, and, therefore, methods for preventing, inhibiting or eradicating Pseudomonal infection are provided, including subjects having cystic fibrosis. In general, these discoveries have led to methods and products using fragments of the lipopolysaccharide of
P. aeruginosa
and using fragments of the CFTR in the manufacture of pharmaceutical products, diagnostic products, research tools, and methods relating hereto.
According to one aspect of the invention, a method for upregulating CFTR expression in the tissue of a subject is provided. A CFTR expression regulator is administered to a subject in need of upregulation of CFTR expression, in an amount effective to increase CFTR expression in the tissue of the subject. The CFTR expression regulator is an isolated polysaccharide that is an LPS core moiety comprising
wherein X is selected from the group consisting of glucose, glucose-rhamnose and H; Y is selected from the group consisting of rhamnose and H; and Z is selected from the group consisting of glucose and H.
A preferred polysaccharide is an LPS core moiety comprising
One particularly useful polysaccharide according to the invention comprises
Another particularly useful polysaccharide according to the invention is:
The foregoing preferred molecules can be isolated CFTR receptor-binding fragments of lipopolysaccharides of
P. aeruginosa.
In one embodiment of the invention, the subject has a condition predisposing the subject to Pseudomonal infection. In another embodiment of the invention, the subject has a Pseudomonal infection. In one important embodiment of the invention, the subject has a defective cystic fibrosis transmembrane conductance regulator gene.
According to another aspect of the invention, a pharmaceutical preparation is provided. The pharmaceutical preparation is a pharmaceutically acceptable carrier and a pharmaceutically effective amount of a CFTR expression regulator. The CFTR expression regulator is as described above. The pharmaceutical preparation can be sterile and can be formulated in a unit dosage in an amount effective for treating Pseudomonal infection. As used herein, “treating” means preventing the onset of, slowing the progression of, or eradicating the existence of the condition being treated, such as a Pseudomonal infection. The pharmaceutical preparation can be formulated as any suitable preparation, including a preparation suitable for inhalation or a preparation suitable for injection.
According to another aspect of the invention, compositions of matter are provided. The compositions are covalent conjugates. One composition is a covalent conjugate of a lipid biocompatible with a human subject and a polysaccharide. The polysaccharide is as described above. In one embodiment, the lipid portion of the conjugate is inserted within the wall of a liposome and the polysaccharide is exposed on the surface of the liposome. The liposome contains a bioactive agent.
Another composition is a covalent conjugate of a bioactive agent and a polysaccharide. Again, the polysaccharide is as described above.
The foregoing covalent conjugates are useful in delivering bioactive agents to cells and/or tissues expressing a CFTR. Thus, methods are provided for delivering a bioactive agent to a tissue expressing a cystic fibrosis transmembrane conductance regulator to treat a condition susceptible to treatment by the bioactive agent. A bioactive agent coupled to a polysaccharide is administered to a subject in need of such treatment, in an amount effective for treating the condition. The polysaccharide is as described above. The bioactive agent can be noncovalently or covalently linked to the polysaccharide, or the bioactive agent can be contained in a liposome comprising a lipid biocompatible with a human subject, wherein the polysaccharide is co
Feisee Lila
Lazar-Wesley Eliane
The Brigham and Women's Hospital Inc.
Wolf Greenfield & Sacks P.C.
LandOfFree
Methods and products for treating pseudomonas infection does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Methods and products for treating pseudomonas infection, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Methods and products for treating pseudomonas infection will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2510995