TECHNICAL FIELD

The invention relates to methods for preventing/treating type-1 diabetes (T1D) and its complications using antibodies (including both endogenous and exogenous) that bind to alpha (α) subunit or beta (β) subunit, or both, of the (Na++K+)-ATPase (NKA) and increase NKA activity. Antibody capable of increasing NKA activity (activation of NKA) is called NKA agonist or NKA activator antibody. In addition, T1D and its complications can also be prevented and treated by administration of an antigenic NKA α or β subunits peptides that induce the production of endogenous antibodies, which specifically bind to the α or β subunits of NKA.

BACKGROUND OF INVENTION

T1D known as juvenile diabetes or insulin-dependent diabetes that destroys insulin-producing beta (β) cells in the pancreatic islets. Insulin is an important hormone needed to convert sugars, starches and other foods into energy. When too little insulin is given, blood sugar levels rise. Over time, this can cause dangerous complications.

T1D causes devastating complications that can damage major organs in the body, including heart, blood vessels, nerves, foot, skin, mouth, eyes and kidneys. Eventually, diabetes complications may be disabling or even life-threatening. Clearly, there is an unmet need for prevention and treatment of T1D and its devastating complications.

NKA is a transmembrane enzyme responsible for the active reciprocal transport of Na+and K+ions across the plasma membrane of all animal cells. NKA comprises two basic subunits: the α subunit and the β subunit. The larger α subunit is the functional subunit, which catalyzes the hydrolysis of ATP for active transport of Na+and K+ions across the plasma membrane; the smaller β subunit does not participate in the catalytic process of the enzyme, but instead acts as a specific chaperone that assists the biogenesis and correct membrane insertion of newly synthesized NKA. The α subunit of NKA has three isoforms including α1, α2 and α3. The β subunit of NKA also has three isoforms including β1, β2 and β3. NKA plays a vitally important role in cell function. Studies have demonstrated that significant reduced NKA activity is tightly associated with diabetes-induced complications including kidney failure and heart failure, demonstrating that protecting and maintaining NKA functional activity from injury is an essential new target for prevention and treatment of T1D.

BRIEF SUMMARY OF INVENTION

The present invention is based on the surprising discovery that NKA antibody agonists or activators with α or β subunit activation site binding specificity can be used to protect insulin-producing β cell function, inhibit hyperglycemia, prevent and treat hyperglycemia caused kidney failure, heart failure, protein damages and other dangerous complications in a subject. Such NKA activator antibodies thus form the basis of methods of treating or preventing T1D associated with diseases.

T1D associated diseases involve but not limited to 1) heart and blood vessel disease and various cardiovascular problems, including coronary artery disease with chest pain (angina), heart attack, heart failure, stroke, atherosclerosis and high blood pressure; 2) nerve damage (neuropathy) in the walls of the blood vessels (capillaries) that nourish the nerves, especially in the legs causing tingling, numbness, burning or pain, hyperglycemia, lose sense of feeling in the affected limbs, nausea, vomiting, diarrhea or constipation; 3) kidney function damage (nephropathy), kidney failure or irreversible end-stage kidney disease; 4) eye damage, damaged the blood vessels of the retina leading to diabetic retinopathy, cataracts and glaucoma; 5) various foot complications; 6) skin and mouth infections and Gum diseases; 7) pregnancy complications may lead to miscarriage, stillbirth, birth defects, diabetic ketoacidosis, retinopathy, pregnancy-induced high blood pressure and preeclampsia or other disease or condition wherein inhibition of T1D would be desirable or necessary.

Examples of NKA activator antibodies having α or β subunit binding specificity that can be used in the methods of the present invention include, but are not limited to, SSA78 (also referred as Jianye 2), SSA401 (also referred as KX-2), SSA412 (also referred as KX-1), JY2948, and JY421228 polyclonal, monoclonal, humanized and human versions thereof, and fragments thereof. These antibodies are capable of increasing NKA enzymatic activity (activation of NKA), which are described in Patent Publication No. PCT/US2006/012912 and U.S. Pat. Nos. 9,974,842, 9,956,275, 9,790,270, 9,527,923, 9,409,949, 9,416,159, 9,238,695, 9,279,020, 9,040,046, 8,945,555, 8,496,929, 8,435,519, 8,383,111, 7,754,210, 10,053,505, 10,214,583 and 1,028,736, which are herein incorporated by reference in its entirety for all purposes.

In addition, T1D and its complications can also be treated by administration of antigenic peptide antigens (or peptide vaccine) of NKA α or β subunits that induce production of endogenous specific antibody activators against the α or β subunits of NKA and increase NKA activity. Such peptides and the administration thereof are also described in U.S. Pat. Nos. 9,974,842, 9,956,275, 9,790,270, 9,527,923, 9,409,949, 9,416,159, 9,238,695, 9,279,020, 9,040,046, 8,945,555, 8,496,929, 8,435,519, 8,383,111, 7,754,210, 10,053,505, 10,214,583 and 10,287,361, which are herein incorporated by reference in its entirety for all purposes. In a first aspect, the invention thus provides methods for protecting insulin-generating β cell function comprising contacting pancreatic β cell with an antibody having binding specificity for the α or β subunit of NKA. Exemplary antibodies that may be used in these methods include, but are not limited to, (i) antibodies having binding specificity for the α or β subunit of NKA, (ii) antibodies having binding specificity for one or more of the peptides represented by SEQ ID NOs:1-7, (iii) NKA activator antibodies in a humanized or human versions thereof, or a fragment or derivative thereof, and (iv) antigens of SEQ ID NOs: 1-7 to generating endogenous NKA activator antibody, in a human or polyclonal versions thereof, or a fragment or derivative thereof. The method may be conducted in vitro or in vivo. The method may also be conducted in blood ex vivo.

In a second aspect, the invention provides methods for preventing autoimmune attacks that destroys the β cells in the pancreas comprising contacting β cells with an antibody having binding specificity for the α or β subunit of NKA. Exemplary antibodies that may be used in these methods include, but are not limited to, (i) antibodies having binding specificity for the α or β subunit of NKA, (ii) antibodies having binding specificity for one or more of the peptides represented by SEQ ID NOs:1-7, (iii) NKA activator antibodies in a humanized or human versions thereof, or a fragment or derivative thereof, and (iv) using SEQ ID NOs: 1-7 peptide antigen to generate endogenous NKA activator antibodies, in human or polyclonal versions thereof, or a fragment or derivative thereof. The method may be conducted in vitro or in vivo. The method may also be conducted in blood ex vivo.

In a third aspect, the invention provides methods for inhibiting hyperglycemia in a subject comprising administering an effective amount of an antibody having binding specificity for the α or β subunit of NKA to a subject in need thereof. Exemplary antibodies that may be used in these methods include, but are not limited to, (i) antibodies having binding specificity for the α or β subunit of NKA, (ii) antibodies having binding specificity for one or more of the peptides represented by SEQ ID NOs:1-7, (iii) NKA activator antibodies in a humanized or human versions thereof, or a fragment or derivative thereof, and (iv) using SEQ ID NOs: 1-7 peptide antigen to generate endogenous NKA activator antibodies, in human or polyclonal versions thereof, or a fragment or derivative thereof. The subject may be one that is characterized has having or at being at greater risk than the general population for one or more of the following diseases and conditions: 1) heart and blood vessel disease and various cardiovascular problems, including coronary artery disease with chest pain (angina), heart attack, heart failure, stroke, atherosclerosis and high blood pressure; 2) nerve damage (neuropathy) in the walls of the blood vessels (capillaries) that nourish the nerves, especially in the legs causing tingling, numbness, burning or pain, hyperglycemia, lose sense of feeling in the affected limbs, nausea, vomiting, diarrhea or constipation; 3) kidney function damage (nephropathy), kidney failure or irreversible end-stage kidney disease; 4) eye damage, damaged the blood vessels of the retina leading to diabetic retinopathy, cataracts and glaucoma; 5) various foot complications; 6) skin and mouth infections and Gum diseases; 7) pregnancy complications may lead to miscarriage, stillbirth, birth defects, diabetic ketoacidosis, retinopathy, pregnancy-induced high blood pressure and preeclampsia or other disease or condition wherein inhibition of T1D would be desirable or necessary.

In a fourth aspect, the invention provides methods for protecting kidney function and preventing diabetic kidney failure in a subject comprising administering an effective amount of an antibody having binding specificity for the α or β subunit of NKA to a subject in need thereof. Exemplary antibodies that may be used in these methods include, but are not limited to, (i) antibodies having binding specificity for the α or β subunit of NKA, (ii) antibodies having binding specificity for one or more of the peptides represented by SEQ ID NOs:1-5, (iii) NKA activator antibodies in a humanized or human versions thereof, or a fragment or derivative thereof, and (iv) using SEQ ID NOs: 1-5 peptide antigen to generate endogenous NKA activator antibodies, in human or polyclonal versions thereof, or a fragment or derivative thereof. The subject may one that is at greater risk than the general population for kidney failure. The subject may be one that is characterized has having or at being at greater risk than the general population for one or more of the following diseases and conditions: diabetic nephropathy, kidney necrosis, kidney failure, irreversible end-stage kidney disease, which requires dialysis or a kidney transplant.

In a fifth aspect, the invention provides methods for treating T1D induced hyperglycemia and complications in a subject comprising administering an effective amount of an antibody having binding specificity for the α subunit of NKA to a subject in need thereof. Exemplary antibodies that may be used in these methods include, but are not limited to, (i) antibodies having binding specificity for the α subunit of NKA, (ii) antibodies having binding specificity for one or more of the peptides antigen or peptide vaccine represented by SEQ ID NOs:1-7, (iii) NKA activator antibodies in a humanized or human versions thereof, or a fragment or derivative thereof, and (iv) using SEQ ID NOs: 1-7 peptide antigen to generate endogenous NKA activator antibodies, in human or polyclonal versions thereof, or a fragment or derivative thereof. The subject may be one that is characterized has having or at being at greater risk than the general population for one or more of the following diseases and conditions: 1) heart and blood vessel disease and various cardiovascular problems, including coronary artery disease with chest pain (angina), heart attack, heart failure, stroke, atherosclerosis and high blood pressure; 2) nerve damage (neuropathy) in the walls of the blood vessels (capillaries) that nourish the nerves, especially in the legs causing tingling, numbness, burning or pain, hyperglycemia, lose sense of feeling in the affected limbs, nausea, vomiting, diarrhea or constipation; 3) kidney function damage (nephropathy), kidney failure or irreversible end-stage kidney disease; 4) eye damage, damaged the blood vessels of the retina leading to diabetic retinopathy, cataracts and glaucoma; 5) various foot complications; 6) skin and mouth infections and Gum diseases; 7) pregnancy complications may lead to miscarriage, stillbirth, birth defects, diabetic ketoacidosis, retinopathy, pregnancy-induced high blood pressure and preeclampsia or other disease or condition wherein inhibition of T1D would be desirable or necessary.

In a sixth aspect, the invention provides methods for treating autoimmune attacks that destroys the β cells in the pancreas in a subject comprising administering an effective amount of an antibody having binding specificity for the α or β subunit of NKA to a subject in need thereof. Exemplary antibodies that may be used in these methods include, but are not limited to, (i) antibodies having binding specificity for the α or β subunit of NKA, (ii) antibodies having binding specificity for one or more of the peptides represented by SEQ ID NOs:1-7, (iii) NKA activator antibodies in a humanized or human versions thereof, or a fragment or derivative thereof, and (iv) using SEQ ID NOs: 1-7 peptide antigen to generate endogenous NKA activator antibodies, in human or polyclonal versions thereof, or a fragment or derivative thereof. The subject may be one that is characterized has having or at being at greater risk than the general population for one or more of the following diseases and conditions: 1) heart and blood vessel disease and various cardiovascular problems, including coronary artery disease with chest pain (angina), heart attack, heart failure, stroke, atherosclerosis and high blood pressure; 2) nerve damage (neuropathy) in the walls of the blood vessels (capillaries) that nourish the nerves, especially in the legs causing tingling, numbness, burning or pain, hyperglycemia, lose sense of feeling in the affected limbs, nausea, vomiting, diarrhea or constipation; 3) kidney function damage (nephropathy), kidney failure or irreversible end-stage kidney disease; 4) eye damage, damaged the blood vessels of the retina leading to diabetic retinopathy, cataracts and glaucoma; 5) various foot complications; 6) skin and mouth infections and Gum diseases; 7) pregnancy complications may lead to miscarriage, stillbirth, birth defects, diabetic ketoacidosis, retinopathy, pregnancy-induced high blood pressure and preeclampsia or other disease or condition wherein inhibition of T1D would be desirable or necessary.

In a seventh aspect, the invention provides methods for preventing T1D induced heart failure and kidney failure in a subject comprising administering an effective amount of an antibody having binding specificity for the α or β subunit of NKA to a subject in need thereof. Exemplary antibodies that may be used in these methods include, but are not limited to, (i) antibodies having binding specificity for the α or β subunit of NKA, (ii) antibodies having binding specificity for one or more of the peptides represented by SEQ ID NOs:1-7, (iii) NKA activator antibodies in a humanized or human versions thereof, or a fragment or derivative thereof, and (iv) using SEQ ID NOs: 1-7 peptide antigen to generate endogenous NKA activator antibodies, in human or polyclonal or monoclonal versions thereof, or a fragment or derivative thereof. Exemplary diseases include, but are not limited to, heart and blood vessel disease and various cardiovascular problems, including coronary artery disease with chest pain (angina), heart failure, heart attack, stroke, atherosclerosis and high blood pressure; diabetic nephropathy, kidney failure or irreversible end-stage kidney disease, which requires dialysis or a kidney transplant.

In an eighth aspect, the invention provides methods for prolonging life span for T1D patient comprising administering an effective amount of an antibody having binding specificity for the α or β subunit of NKA to a subject in need thereof. Exemplary antibodies that may be used in these methods include, but are not limited to, (i) antibodies having binding specificity for the α or β subunit of NKA, (ii) antibodies having binding specificity for one or more of the peptides represented by SEQ ID NOs:1-7, (iii) NKA activator antibodies in a humanized or human versions thereof, or a fragment or derivative thereof, and (iv) using SEQ ID NOs: 1-7 peptide antigen to generate endogenous NKA activator antibodies, in human or polyclonal or monoclonal versions thereof, or a fragment or derivative thereof.

In each of these aspects, the antibody may be in a pharmaceutical formulation comprising the antibody and a pharmaceutically acceptable carrier.

The foregoing has outlined rather broadly the features and technical advantages of the present invention in order that the detailed description of the invention that follows may be better understood. Additional features and advantages of the invention will be described herein, which form the subject matter of the claims of the invention.

