Anti-marinobufagenin antibodies and methods for their use
Inventors
Bagrov, Alexei • Fedorova, Olga V. • Lakatta, Edward G. • Simbirtsev, Andrey • Kotov, Alexander • Kolodkin, Nikolai
Assignees
US Department of Health and Human Services
Publication Number
US-8038997-B2
Publication Date
2011-10-18
Expiration Date
2026-06-26
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Abstract
Described herein are deposited hybridoma cell lines and the monoclonal antibodies produced by these hybridomas, and antigen binding fragments thereof. These monoclonal antibodies and antigen binding fragments specifically bind marinobufagenin. The disclosure also encompasses the use of these monoclonal antibodies or antigen binding fragments in a method for detecting the presence of marinobufagenin in a biological sample. Also provided are methods for the use of these monoclonal antibodies or antigen binding fragments as prophylactic, therapeutic, and diagnostic agents for the detection, inhibition and treatment of a cardiovascular disease, for example, essential hypertension, hypertension associated with preeclampsia, eclampsia, or renal failure, or myocardial fibrosis in a subject.
Core Innovation
The invention describes hybridoma cell lines and monoclonal antibodies they produce, along with antigen binding fragments that specifically bind marinobufagenin (MBG). These monoclonal antibodies and fragments are employed to detect MBG presence in biological samples and can be used as prophylactic, therapeutic, and diagnostic agents against cardiovascular diseases related to elevated MBG levels, including hypertension, myocardial fibrosis, preeclampsia, eclampsia, and uremic cardiomyopathy.
The problem addressed arises from the role of elevated digitalis-like sodium pump ligands (SPLs), such as MBG, in cardiovascular conditions. Hypertension, particularly essential hypertension and hypertension associated with pregnancy complications like preeclampsia and eclampsia, lacks effective targeted therapies. Likewise, chronic renal failure patients develop uremic cardiomyopathy marked by cardiac hypertrophy and fibrosis, partially linked to elevated steroid molecules like MBG that inhibit Na+/K+-ATPase activity. Detecting and neutralizing MBG can thus mitigate vasoconstriction and hypertension associated with these diseases.
Claims Coverage
The patent includes 56 inventive features primarily covering hybridoma cell lines, monoclonal and humanized antibodies specific to marinobufagenin, diagnostic methods, and therapeutic methods using these antibodies.
Hybridoma cell lines producing anti-marinobufagenin antibodies
Hybridoma cell lines deposited under ATCC accession numbers PTA-6724, PTA-6725, and PTA-6788 that produce monoclonal antibodies specifically binding marinobufagenin.
Monoclonal antibodies and antigen-binding fragments that specifically bind marinobufagenin
Monoclonal antibodies and their MBG-binding fragments produced by the deposited hybridomas, which can be labeled with detectable agents like enzymes, fluorochromes, haptens, radioisotopes, or electron-dense compounds.
Diagnostic methods using specific monoclonal antibodies to detect MBG
Methods of detecting MBG in biological samples by contacting the sample with the specific monoclonal antibodies or fragments, optionally labeled, and detecting antibody binding. Additionally, methods use a second antibody binding the monoclonal antibody for detection.
Methods of diagnosing cardiovascular diseases associated with increased MBG
Methods of diagnosing cardiovascular diseases such as hypertension, essential hypertension, preeclampsia, eclampsia, myocardial fibrosis, and uremic cardiomyopathy by contacting biological samples with the specific monoclonal antibodies and detecting increased MBG levels.
Therapeutic methods of treating cardiovascular disease by administering anti-MBG monoclonal antibodies
Methods of treating or inhibiting cardiovascular diseases associated with elevated MBG by administering a therapeutically effective amount of monoclonal antibodies produced by the hybridomas in a pharmaceutically acceptable carrier, including combination with other cardiovascular drugs.
Humanized antibodies comprising CDRs from hybridoma-produced antibodies
Humanized antibodies specific for marinobufagenin that include the complementarity determining regions of monoclonal antibodies produced by the deposited hybridomas and a human antibody framework region.
Compositions containing the monoclonal antibodies or hybridoma cell lines
Pharmaceutical and other compositions comprising the monoclonal antibodies or the hybridoma cell lines, optionally in carriers such as saline.
Substrates or supports comprising the monoclonal antibody
Substrates, for example beads, that have the monoclonal antibodies adsorbed or conjugated for purposes such as detection or purification of MBG.
Methods of reducing arterial blood pressure and increasing Na+/K+-ATPase activity by administering monoclonal antibodies
Methods employing administration of therapeutically effective amounts of the monoclonal antibodies to reduce arterial blood pressure and increase Na+/K+-ATPase activity in subjects with hypertension and related cardiovascular conditions, including preeclampsia, myocardial fibrosis, and uremic cardiomyopathy.
The claims comprehensively cover the deposited hybridoma cell lines producing monoclonal antibodies binding MBG, their antibody fragments, labeling for detection, methods for diagnosing MBG-related cardiovascular diseases, therapeutic use of these antibodies, humanized variants, compositions and substrates containing the antibodies, and methods of reducing blood pressure and enhancing Na+/K+-ATPase activity with these antibodies.
Stated Advantages
The monoclonal antibodies are more effective than DIGIBIND in restoring Na+/K+-ATPase activity in erythrocytes from preeclampsia patients.
Administration of the monoclonal antibodies reduces systolic blood pressure in hypertensive animal models including Dahl salt-sensitive rats and pregnancy-associated hypertension models.
The antibodies can detect MBG specifically in biological samples with high specificity and cross-reactivity minimized against other steroids.
Treatment with anti-MBG antibodies attenuates cardiac hypertrophy, oxidant stress, and myocardial fibrosis in animal models of uremic cardiomyopathy and angiotensin II-induced hypertension.
The monoclonal antibodies enable diagnostic accuracy and facilitate treatment decisions for diseases associated with elevated MBG levels.
Documented Applications
Diagnostic use for detecting the presence of marinobufagenin in biological samples such as blood, serum, urine, or plasma to assess cardiovascular disease risk or presence.
Diagnostic methods for hypertension, essential hypertension, preeclampsia, eclampsia, uremic cardiomyopathy, and myocardial fibrosis by detecting elevated MBG levels using the described antibodies.
Therapeutic methods to treat or inhibit cardiovascular diseases associated with increased MBG levels by administering anti-MBG monoclonal antibodies to human or mammalian subjects.
Use of anti-MBG antibodies to reduce blood pressure and restore Na+/K+-ATPase activity in subjects with hypertension or cardiovascular conditions.
Use of anti-MBG antibodies to attenuate myocardial fibrosis and pathological remodeling in angiotensin II-induced hypertension animal models.
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