Drug – bio-affecting and body treating compositions – Enzyme or coenzyme containing – Hydrolases
Reexamination Certificate
2002-06-03
2003-07-15
Prouty, Rebecca E. (Department: 1652)
Drug, bio-affecting and body treating compositions
Enzyme or coenzyme containing
Hydrolases
C514S002600, C514S015800
Reexamination Certificate
active
06592865
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to polypeptides that regulate production of Angiotensin 1-9 via such mechanisms as, for example, inhibition of ACE-2. Such polypeptides have uses for example, in the detection, isolation, and/or purification of ACE-2 and/or Angiotensin 1-9. The invention also relates to nucleic acid molecules encoding these ACE-2 binding polypeptides, vectors and host cells containing these nucleic acids, and methods for producing the same. The present invention also relates to methods and compositions for detecting, diagnosing, or prognosing a disease or disorder associated with aberrant ACE-2 or Angiotensin 1-9 expression or inappropriate function of ACE-2.or Angiotensin 1-9, comprising ACE-2 binding polypeptides or fragments or variants thereof, that specifically regulate ACE-2 action. The present invention further relates to methods and compositions for preventing, treating or ameliorating a disease or disorder associated with aberrant ACE-2 or Angiotensin 1-9 expression or inappropriate ACE-2 function or Angiotensin 1-9 function, comprising administering to an animal, preferably a human, an effective amount of one or more ACE-2 binding polypeptides or fragments or variants thereof, that specifically regulate ACE-2.
BACKGROUND OF THE INVENTION
The renin-angiotensin system (RAS) plays an important role in circulatory homeostasis at both systemic and local levels. Angiotensin converting enzyme (ACE), a 175 kD protein known to be widely distributed throughout the cardiovascular system, has been long recognized as the key enzyme in the generation of angiotensin II, a peptide that regulates fluid balance, blood pressure and local blood flow in a number of tissues (Peach, M. J.
Physiological Reviews
57:313-370 (1997)). As part of an ongoing strategy to establish genes associated with cardiovascular function via high throughput cDNA sequencing, we identified a member of the RAS family of enzymes, ACE-2, from human kidney. This enzyme was also identified in a variety of tissues by others (Donoghue et al.,
Circulation Research
87:e1-e9 (2000), Tipinis et al.,
The Journal of Biological Chemistry
275:33238-33243 (2000)). The unmodified ACE-2 protein contains transmembrane and signal peptide domains, but unlike ACE, ACE-2 contains just one single extracellular Zn
+2
binding metalloprotease domain (Tipinis et al.,
The Journal of Biological Chemistry
275:33238-33243 (2000)). ACE-2 mRNA has a more limited expression pattern than ACE (Donoghue et al.,
Circulation Research
87:e1-e9 (2000)) and, remarkably, no detectable expression in lungs (unpublished data).
ACE-2 and related carboxypeptidases (Snyder et al.,
The Journal of Biological Chemistry
260:7857-7860 (1985); Kokkonen et al.,
Circulation
95:1455-1463 (1997)) catalyze the removal of the C-terminal leucine from angiotensin I to form the nonapeptide angiotensin 1-9 (A1-9) (SEQ ID NO:145) or des-Leu
10
-angiotensin I (Donoghue et al.,
Circulation Research
87:e1-e9 (2000); Tipinis et al.,
The Journal of Biological Chemistry
260:7857-7860 (2000); Snyder et al.,
The Journal of Biological Chemistry
260: 7857-7860 (1985); Snyder et al.,
Biochemica et Biophysica Acta
871:1-5 (1986)). Circulating A1-9 has been detected in vivo at levels twice that of angiotensin II (Oparil et al.,
Circulation Research
29 682-690 (1971); Johnson et al.,
Peptides
10:489-492) (1989)). In the case of ACE-2, the above reaction is not blocked by captopril, lisinopril or enalaprilat (Donoghue et al.,
Circulation Research
87:e1-e9 (2000); Tipinis et al.,
The Journal of Biological Chemistry
275:33238-33243 (2000)). The unique expression profile of ACE-2, spectum of its enzymatic activity and inhibitory effects of its product A1-9 on ACE have led to the speculation that ACE-2 functions to affect circulatory homeostasis by promoting vasodilation (Donoghue et al.,
Circulation Research
87:e1-e9 (2000); Snyder et al.,
The Journal of Biological Chemistry
260:7857-7860 (1985)). However, A1-9 has been shown to be a weak vasoconstrictor in isolated rat aorta and have weak pressor activity in anesthetized rats and dogs (Oparil et al.,
Circulation Research
29:682-690 (1971)). Therefore, we hypothesized that one of the physiologic roles of ACE-2 is to increase arterial pressure through the actions of its catabolic product, A1-9. As such, ACE-2 might be a valid target for drug development in hypertension.
Accordingly, molecules that specifically bind ACE-2 would find a variety of uses in the study of ACE-2, angiotensin 1-9) (SEQ ID NO:145), and angiotensin, as well as ACE, and its known substrates: Angiotensin II (SEQ ID NO:144), Angiotensin 1-7, des-Asp, bradykinin, neurotensin, and Substance P. Further, molecules that specifically bind ACE-2 would also find a variety of uses in the manufacture and purification of ACE-2, ACE, angiotensin, angiotensin II, and/or Angiotensin 1-9 in commercial and medically pure quantities, and in the development new therapeutic or diagnostic reagents. ACE-2 binding polypeptides may also find medical utility in, for example, the treatment of cardiovascular disorders (e.g., hypertension, chronic heart failure, left ventricular failure, stroke, cerebral vasospasm after subarachnoid injury, atherosclerotic heart disease, and retinal hemorrhage), renal disorders (e.g., renal vein thrombosis, kidney infarction, renal artery embolism, renal artery stenosis, myocardial hypertrophy, hypertrophy and/or hyperplasia of conduit and/or resistance vessels, myocyte hypertrophy, and fibroblast proliferative diseases), inflammatory diseases (e.g., SIRS (systemic Inflammatory Response Syndromes), sepsis, polytrauma, inflammatory bowl disease, acute and chronic pain, rheumatoid arthritis, and osteo arthritis), allergic disorders (e.g., asthma, adult respiratory distress syndrome, wound healing, and scar formation), as well as several other disorders and/or diseases (e.g., periodontal disease, dysmenorrhea, premature labor, brain edema following focal injury, diffuse axonal injury, and reperfusion injury).
SUMMARY OF THE INVENTION
The present invention provides new polypeptides and families of polypeptides that specifically bind ACE-2 and/or ACE-2-like polypeptides. In particular, the invention encompasses polypeptides that specifically bind to a polypeptide or polypeptide fragment of human ACE-2 (SEQ ID NOs:138 and/or 142).
In particular, the invention relates to ACE-2 binding polypeptides comprising, or alternatively consisting of, an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-136, preferably SEQ ID NOs: 11-39, more preferably SEQ ID NOs:23-24 and 36-39, as referred to below, in Tables 1-2 and Example 1 below, and fragments and variants thereof.
In specific preferred embodiments, the ACE-2 binding polypeptides of the invention bind ACE-2 and/or ACE-2-like polypeptides with high affinity. In other embodiments, the ACE-2 binding polypeptides of the invention reversibly bind ACE-2 and/or ACE-2-like polypeptides. In still other embodiments, the ACE-2 binding polypeptides of the invention irreversibly bind ACE-2 and/or ACE-2-like polypeptides.
The cysteine residues in certain polypeptides according to the invention are believed to form a disulfide bond, which would cause the polypeptide containing these cysteine residues to form a stable loop structure under non-reducing conditions. Especially preferred ACE-2 binding polypeptides of the invention are polypeptide molecules that comprise amino acid sequences that form stable loop structures or other stable structures that bind ACE-2 or ACE-2-like polypeptides.
Preferred binding polypeptides specific for ACE-2 include two separated, invariant cyteine residues and are thus capable of forming a cyclic strucure under non-reducing conditions via a disulfide bond formed between the cysteine side chains. Specific ACE-2 binding polypeptides according to the present invention include polypeptides comprising amino acid sequences of the following general formulae I-X:
Z
1
-X
1
-A-X
2
-X
3
-C-X
4
-X
5
-F-Z
2
(SEQ ID NO:1) &
Parry Tom J.
Sekut Les
Human Genome Sciences Inc.
Prouty Rebecca E.
Swope Sheridan L.
LandOfFree
Methods and compositions for modulating ACE-2 activity 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 compositions for modulating ACE-2 activity, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Methods and compositions for modulating ACE-2 activity will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3078055