Methods of treating hypertension with metabolic syndrome using MANP

Abstract

The present disclosure provides methods and materials related to MANP. In one aspect, the disclosure provides a method of treating hypertension with metabolic syndrome in a patient in need thereof by administering MANP. In one aspect, the disclosure provides methods comprising administering MANP to a patient to reduce blood pressure (BP). In one aspect, the methods of the present disclosure are useful for treating patients with low baseline cGMP. In one aspect, the disclosure provides methods of treatment using MANP to increase plasma cGMP, plasma ANP-like peptide, insulin sensitivity, or plasma NEFA. In one aspect, the disclosure provides methods of treatment using MANP to reduce plasma glucose levels.

Description

[0001] Related Applications

[0002] This application claims priority to U.S. Provisional Application Serial No. 63 / 486,421, filed February 22, 2023, the entirety of which is incorporated by reference herein

[0003] Statement as to Federally Sponsored Research

[0004] This application was made with government support under HL136340 and HL134668 awarded by the National Institutes of Health. The government has certain rights in the application. TECHNICAL FIELD

[0005] The present disclosure provides methods for treating hypertension with metabolic syndrome in a patient using alternatively spliced atrial natriuretic peptide (MANP). BACKGROUND

[0006] Hypertension, also known as high blood pressure (BP), is a long-term medical condition in which the BP within the arteries is elevated. Metabolic syndrome is a cluster of cardiovascular and metabolic risk factors that predispose patients to major cardiovascular disease. Hypertension is one of the main clinical features of metabolic syndrome. Indeed, hypertension is present in 77% of patients affected by metabolic syndrome, and conversely, visceral obesity is a key risk factor for the development of hypertension. Both hypertension and metabolic syndrome are important risk factors for cardiovascular disease individually; when they coexist, the risk is doubled. Despite this interrelated pathological state, none of the currently available anti-hypertensive drugs have been reported to produce favorable metabolic effects, and no therapy has been specifically approved for the treatment of hypertension associated with metabolic syndrome.

[0007] The difficulty in treating hypertension with metabolic syndrome can be attributed to the different responses of different patients to potential treatments. The lack of objective and reliable predictors of treatment outcome is one of the many reasons for the lack of drugs approved for the treatment of hypertension with metabolic syndrome. While published studies (see, e.g., Chen et al. (2021), Hypertension, 78(6): 1859-1867) demonstrate that MANP can be administered to modulate certain symptoms of resistant hypertension, there are no known predictors that can reliably predict a patient’s response to any hypertension treatment in the context of treating hypertension with metabolic syndrome. There remains a need for treatment methods that employ reliable predictors to deliver effective treatments to patients with hypertension with metabolic syndrome. SUMMARY

[0008] The present disclosure includes methods and materials related to MANPs. The present disclosure also includes methods of treating hypertension with metabolic syndrome in a patient in need thereof having a low baseline plasma cyclic guanosine monophosphate (cGMP) by administering a MANP to the patient. In one aspect, the present disclosure includes administering a MANP to a patient in need thereof to increase plasma cGMP, to reduce blood pressure, or to increase plasma atrial natriuretic peptide (ANP)-like peptides in the patient. In one aspect, the present disclosure also includes administering a MANP to reduce plasma glucose levels, to increase insulin sensitivity, or to increase plasma non-esterified fatty acids (NEFA) in the patient.

[0009] In one aspect, the present disclosure provides and includes a method for treating hypertension with metabolic syndrome in a patient in need thereof having a baseline plasma cGMP of less than 10 pmol / mL. The method of the present disclosure includes increasing plasma cGMP compared to baseline by administering an effective dose of a MANP to the patient. In one aspect, the present disclosure provides and includes a method of reducing BP in a patient in need thereof having a baseline plasma cGMP of less than 10 pmol / mL. The method of the present disclosure includes increasing plasma cGMP compared to baseline by administering an effective dose of a MANP to the patient. In one aspect, the present disclosure provides and includes a method of increasing plasma ANP-like peptides in a patient in need thereof having a baseline plasma cGMP of less than 10 pmol / mL. The method of the present disclosure includes increasing plasma cGMP compared to baseline by administering an effective dose of a MANP to the patient. In one aspect, the present disclosure provides and includes a method of reducing plasma glucose levels in a patient in need thereof comprising administering an effective dose of a MANP to the patient. In one aspect, the present disclosure provides and includes a method of increasing insulin sensitivity in a patient in need thereof comprising administering an effective dose of a MANP to the patient. In one aspect, the present disclosure provides and includes a method of increasing plasma NEFA in a patient in need thereof comprising administering an effective dose of a MANP to the patient.

[0010] In an aspect, the methods of the present disclosure increase plasma cGMP in a patient in need thereof 2 to 8 pmol / mL from baseline within 0.5 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces sitting systolic BP by 0.1% to 15% compared to baseline sitting systolic BP within 12 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces sitting diastolic BP by 0.1% to 5% compared to baseline sitting diastolic BP within 6 hours after administration of an effective dose of MANP. In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide by 1 to 70 pg / mL compared to baseline plasma ANP-like peptide within 0.5 hours after administration of an effective dose of MANP. In an aspect, a method of increasing insulin sensitivity in a patient in need thereof increases the patient's Homeostatic Model Assessment Sensitivity (HOMA2-S) value by 0.1% to 35% compared to baseline HOMA2-S value within 4 hours after administration of an effective dose of MANP. In an aspect, a method of increasing insulin sensitivity in a patient in need thereof decreases the patient's HOMA2-Resistance (HOMA2-IR) value by 0.1 to 0.6 compared to baseline HOMA2-IR value within 4 hours after administration of an effective dose of MANP. In an aspect, a method of increasing plasma NEFA in a patient in need thereof increases plasma NEFA by 1 μΜ compared to baseline plasma NEFA within 1 hour after administration of an effective dose of MANP.

[0011] In an aspect, the MANP can be administered subcutaneously (e.g., at a dose of about 0.1 μg / kg to 5 μg / kg, or about 6.5 μg to 750 μg). In an aspect, the MANP can be administered intravenously (e.g., at a dose of about 10 pmol / kg / minute to 100 nmol / kg / minute). In an aspect, the MANP can be administered intravenously and subsequently administered subcutaneously. For example, the MANP can be administered intravenously at a dose of about 10 pmol / kg / minute to about 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of about 0.1 μg / kg to 5 μg / kg.

[0012] In an aspect, the patient in need thereof exhibits hypertension despite use of at least one anti-hypertensive medication. In an aspect, the patient in need thereof exhibits a sitting systolic BP of between 140 mmHg and 180 mmHg, or a sitting diastolic BP of between 90 mmHg and 100 mmHg prior to administration of the MANP.

[0013] In an aspect, the patient in need thereof has a body mass index greater than 25 kg / m 2body mass index (BMI). In one aspect, the patient in need thereof is a male patient having a waist circumference greater than or equal to 102 cm. In one aspect, the patient in need thereof is a female patient having a waist circumference greater than or equal to 88 cm. In one aspect, the patient in need thereof is a patient having high serum triglycerides prior to receiving the MANP according to the disclosure. In one aspect, the patient in need thereof is a patient having low HDL cholesterol prior to receiving the MANP according to the disclosure. In one aspect, low HDL cholesterol is identified by HDL cholesterol less than 40 mg / dL in male patients. In one aspect, low HDL cholesterol is identified by HDL cholesterol less than 50 mg / dL in female patients. In one aspect, the patient in need thereof is a patient who is receiving pharmacotherapy for low HDL cholesterol to increase HDL cholesterol. In one aspect, the patient in need thereof is a patient having high fasting plasma glucose measured prior to receiving the MANP according to the disclosure. In one aspect, high fasting plasma glucose is identified by fasting plasma glucose 100 mg / dL or above in the patient.

[0014] In one aspect, the patient in need thereof is not pregnant. In one aspect, the patient in need thereof is not lactating. In one aspect, the patient in need thereof (within 2 years prior to administration of the MANP according to the methods of the disclosure) has no history of alcohol abuse, illicit drug use, severe mental illness, physiological dependence on any opioid drug, or any history of drug abuse or addiction. In one aspect, the patient in need thereof has no history of difficulty donating blood, or has not donated blood or blood products within 45 days prior to administration. In one aspect, the patient in need thereof has no history of coronary artery disease or cerebrovascular disease or syncope. In one aspect, the patient in need thereof has no history of epilepsy or other seizure disorder. In one aspect, the patient in need thereof has no history of organ transplant. In one aspect, the patient in need thereof has no malignancy within 5 years prior to administration of the MANP according to the methods of the disclosure. In one aspect, the patient in need thereof has no history of clinically significant endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery. BRIEF DESCRIPTION OF DRAWINGS

[0015] The application is further disclosed in reference to the drawings, in which:

[0016] Figure 1 schematic structure of a MANP is illustrated.

[0017] Figure 2 a schematic overview of a study protocol to study the effect of a MANP according to the disclosure in subjects having hypertension with metabolic syndrome is illustrated.

[0018] Figure 3AGraph of circulating levels of ANP-like peptide in patients during the 24 hours following subcutaneous administration of MANP according to the present disclosure compared to placebo.

[0019] Figure 3B Graph of change in circulating levels of cGMP during the 24 hours following subcutaneous administration of MANP according to the present disclosure compared to placebo.

[0020] Figure 4A Graph of change in sitting systolic BP during the 24 hours following subcutaneous administration of MANP according to the present disclosure compared to placebo.

[0021] Figure 4B Graph of change in sitting diastolic BP during the 24 hours following subcutaneous administration of MANP according to the present disclosure compared to placebo.

[0022] Figure 4C Graph of change in heart rate during the 24 hours following subcutaneous administration of MANP according to the present disclosure compared to placebo.

[0023] Figure 5A Correlation between baseline characteristics of patients receiving treatment according to the present disclosure and change in mean systolic BP is provided.

[0024] Figure 5B Correlation between baseline characteristics of patients receiving treatment according to the present disclosure and change in mean diastolic BP is provided.

[0025] Figure 6A Graph of maximum ANP-like peptide increase following a single MANP injection in male (n=5) and female (n=12) subjects according to the present disclosure.

[0026] Figure 6B Graph of maximum cGMP increase following a single MANP injection in male (n=5) and female (n=12) subjects according to the present disclosure.

[0027] Figure 6C Graph of change in mean systolic BP following a single MANP injection in male (n=5) and female (n=12) subjects according to the present disclosure.

[0028] Figure 6D Graph of change in mean diastolic BP following a single MANP injection in male (n=5) and female (n=12) subjects according to the present disclosure.

[0029] Figure 7A Correlation between baseline characteristics of patients receiving treatment according to the present disclosure and maximum ANP-like peptide increase is provided.

[0030] Figure 7BA correlation between baseline characteristics of patients receiving treatment according to the present disclosure and maximum cGMP increase is provided.

[0031] Figure 8A A comparison plot of change in HOMA2 insulin sensitivity of patients between baseline and 4 hours after administration of MANP according to the present disclosure compared to placebo is provided.

[0032] Figure 8B A comparison plot of change in HOMA2 insulin resistance of patients between baseline and 4 hours after administration of MANP according to the present disclosure compared to placebo is provided.

[0033] In several views, corresponding reference numbers indicate corresponding parts. The examples set forth herein demonstrate embodiments of the present disclosure and should not be construed as limiting the scope of the present disclosure in any manner. DETAILED DESCRIPTION

[0034] This description is not intended to be a detailed catalog of all the different ways or all the different features that can be implemented in the present disclosure. For example, features illustrated with respect to one embodiment can be incorporated into other embodiments, and features illustrated with respect to a particular embodiment can be deleted from that embodiment. Thus, the present disclosure contemplates, in some aspects, excluding or omitting any feature or combination of features set forth herein. Additionally, the various aspects of the disclosure set forth herein are described and explained only in connection with certain embodiments. However, it should be understood that many variations and modifications of the various aspects of the disclosure can be made in light of the above teachings. Further, the disclosures herein are intended to embrace all alternatives, modifications, and variations that can fall within the scope of the present disclosure. In other instances, well-known structures, interfaces, and processes have not been shown in detail so as not to unnecessarily obscure the disclosure. Nothing in the specification should be construed as implying any special relationship between the inventor and any instrumentality or location mentioned herein. The following description is intended to illustrate some specific aspects of the disclosure, and is not intended to specify all the scope of the disclosure.

[0035] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Also, all publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety for the teachings relevant to the sentence and / or paragraph in which the reference is present. References to techniques employed herein are intended to refer to the techniques as commonly understood in the art, including variations or substitutions of those techniques that would be apparent to one of skill in the art. In case of conflict, the present specification, including definitions, controls.

[0036] All publications, patent applications, patents, and other references cited herein are incorporated by reference in their entireties for the teachings relevant to the sentence and / or paragraph in which the reference is present. References to techniques employed herein are intended to refer to the techniques as commonly understood in the art, including variations or substitutions of those techniques that would be apparent to one of skill in the art. In case of conflict, the present specification, including definitions, controls.

[0037] It is expressly intended that all combinations of the various features and modifications described herein can be employed with any other aspect or feature of the disclosure. In addition, the disclosure contemplates that any feature or combination of features described herein can be excluded from any aspect or feature of the disclosure.

[0038] The methods disclosed herein comprise and encompass one or more steps or actions for accomplishing a described method. The method steps and / or actions can be interchanged with one another without departing from the scope of the disclosure. In other words, unless a particular order of steps or actions is required for proper operation of an embodiment, the order and / or use of specific steps and / or actions can be modified without departing from the scope of the disclosure. Although methods and materials similar or equivalent to those described herein can be used in the practice of aspects of the disclosure, suitable methods and materials are described herein.

[0039] As used in the specification and the appended claims, the singular forms "a," "an" and "the" are intended to include plural forms as well, unless the context clearly indicates otherwise.

[0040] As used herein, "and / or" means and / or includes any and all possible combinations of one or more of the associated listed items, as well as the lack of combination when interpreted in the alternative ("or").

[0041] As used herein, the terms "about" and "approximately" when used in reference to a measurable quantity, such as length, frequency, duration, and the like, means an amount that is nearly correct within a range that is acceptable for practice or implementation of the disclosure. For example, "about" can mean within 20%, 10%, 5%, 1%, 0.5%, or even 0.1% of the stated amount.

[0042] As used herein, phrases such as "between X and Y" and "between about X and Y" should be interpreted to include X and Y. As used herein, phrases such as "between about X and Y" mean "between about X and about Y" and phrases such as "from about X to Y" mean "from about X to about Y."

[0043] As used herein, the term "exemplary" is used in the sense of being an example, instance or illustration. Any aspect or aspect described as "exemplary" is not necessarily to be construed as preferred or advantageous over other aspects. Rather, use of the term "exemplary" is intended to present concepts in a concrete manner and the disclosed subject matter is not limited to such examples. The terms "comprises," "comprising," "includes," "including" and the like can be used herein and are intended to mean that the disclosed subject matter can include certain elements or a combination of elements, but not excluding the presence of one or more other elements or combination of elements.

[0044] The present disclosure provides and includes methods for treating hypertension with metabolic syndrome in a patient. The methods of the present disclosure can include administration of a MANP as provided herein. In an aspect, the methods of treatment according to the present disclosure can beneficially affect or reduce one or more symptoms associated with the condition or one or more underlying causes of the condition. In an aspect, the present disclosure provides and includes methods of reducing BP in a patient in need thereof, comprising administering a MANP to the patient. In an aspect, the present disclosure provides and includes methods of increasing plasma ANP-like peptides in a patient in need thereof, comprising administering a MANP to the patient. In an aspect, the present disclosure provides and includes methods of reducing plasma glucose levels in a patient in need thereof, comprising administering a MANP to the patient. In an aspect, the present disclosure provides and includes methods of increasing insulin sensitivity in a patient in need thereof, comprising administering a MANP to the patient. In an aspect, the present disclosure provides and includes methods of increasing plasma NEFA in a patient in need thereof, comprising administering a MANP to the patient. In an aspect, the present disclosure provides and includes methods for treating a patient with low baseline plasma cGMP. In an aspect, the low baseline plasma cGMP is a plasma cGMP of less than 10 pmol / mL (e.g., less than 5 pmol / mL, less than 1 pmol / mL, less than 0.5 pmol / mL, less than 0.1 pmol / mL, less than 0.05 pmol / mL, or less than 0.01 pmol / mL).

[0045] As used herein, the terms "treat" or "treatment" are methods used to obtain beneficial or desired clinical results. In an aspect, the term "treatment" means administration of a MANP disclosed herein that partially or completely alleviates, ameliorates, relieves, inhibits, delays onset of, reduces severity and / or reduces incidence of one or more symptoms, features or causes of hypertension with metabolic syndrome. The term "treatment" includes administration of a MANP disclosed herein to prevent or delay the onset of symptoms, complications or biochemical indicators of hypertension with metabolic syndrome, to alleviate symptoms, or to stop or slow further progression of hypertension with metabolic syndrome. Treatment can be prophylactic (to prevent or delay the onset of hypertension with metabolic syndrome), or therapeutic to inhibit or alleviate symptoms after hypertension with metabolic syndrome manifestations.

[0046] In an aspect, the present disclosure includes a method of treatment that can beneficially affect or alleviate one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more symptoms associated with a condition. In an aspect, the present disclosure includes a method of treatment that can beneficially affect or alleviate up to five, up to six, up to seven, up to eight, up to nine, up to ten, up to fifteen, or up to twenty symptoms associated with a condition. In an aspect, a method of treatment according to the present disclosure can beneficially affect or alleviate one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more underlying causes of a condition. In an aspect, the present disclosure includes a method of treatment that can beneficially affect or alleviate up to five, up to six, up to seven, up to eight, up to nine, up to ten, up to fifteen, or up to twenty underlying causes of a condition.

[0047] As used herein, “hypertension” refers to a systolic BP of 140 millimeters of mercury (mmHg) to 180 mmHg or a diastolic BP of 90 mmHg to 100 mmHg while sitting. In an aspect, hypertension is a BP reading with a systolic BP of 130 mmHg and a diastolic BP of 80 mmHg or higher.

[0048] In an aspect, BP is measured by a sphygmomanometer or blood pressure gauge. In an aspect, the sphygmomanometer includes an inflatable cuff, a measurement unit (mercury manometer or aneroid manometer), and a mechanism for inflation, which can be a manually operated bulb and valve or an electrically operated pump. In an aspect, the sphygmomanometer includes a manual gauge. In an aspect, the sphygmomanometer includes a digital gauge.

[0049] In an aspect, when measuring seated BP, the subject sits in a comfortable chair with back support and readings are taken at least 5 minutes later. The subject’s feet are flat on the floor and legs are uncrossed. The subject’s arms are placed on a table with the cuff level with the chest.

[0050] In an aspect, systolic and diastolic BP are measured using methods known in the art. In an aspect, the cuff of a sphygmomanometer is placed smoothly and snugly around the upper arm, at approximately the same vertical height as the heart while the subject is seated with the arm supported. In an aspect, the size of the cuff is chosen and fitted to the patient. In an aspect, measurements are taken on both arms to determine if the pressure in one arm is significantly higher than the other. A difference of 10 mm Hg can be a sign of aortic coarctation. If the readings differ between the two arms, the arm with the higher reading is used for subsequent readings. The cuff is inflated until the artery is completely occluded. In an aspect, using a manual instrument, the brachial artery is auscultated with a stethoscope, and the examiner slowly releases the pressure in the cuff at a rate of about 2 mm Hg per heartbeat. As the pressure in the cuff falls, a "whooshing" or thumping sound is heard when blood flow in the artery resumes. The pressure at which this sound begins is noted and recorded as the systolic pressure. The cuff pressure is released further until the sound is no longer heard. This is recorded as the diastolic pressure. In an aspect, a digital instrument uses a cuff that can be placed around the upper arm, wrist, or finger, in all cases elevated to the same height as the heart. The digital instrument inflates and gradually lowers the pressure in the cuff in the same manner as the manual instrument, and measures blood pressure by oscillometry.

[0051] In an aspect, the presence or extent of hypertension can be assessed using methods known in the art, including but not limited to general clinical examination to assess BP, heart rate, heart rhythm, arterial oxygen, and hemoglobin levels; echocardiography to measure ejection fraction, left ventricular (LV) and left atrial (LA) diameters, LV wall motion, LV filling pressure, and diastolic function by pulse and tissue Doppler; measurement of cardiac output, pulmonary capillary wedge pressure, pulmonary artery pressure, right ventricular pressure, right atrial pressure, and systemic and pulmonary vascular resistance using a Swan-Ganz catheter; assessment of kidney function by determination of glomerular filtration rate, serum creatinine, and blood urea nitrogen; and measurement of biomarkers such as B-type natriuretic peptide (BNP), amino-terminal proBNP (NT-proBNP), troponin-T, troponin-I, C-reactive protein (CRP), and creatine kinase, serum cystatin-C, albuminuria, neutrophil gelatinase-associated lipocalin (NGAL), N-acetyl-beta-D-glucosaminidase (NAG), kidney injury molecule 1 (KIM-1), angiotensin-II, renin, aldosterone, and inflammatory cytokines (e.g., interleukin (IL)-6, IL-18, etc.).

[0052] As used herein, "metabolic syndrome" refers to a condition diagnosed by criteria established by the National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ATP III). Specifically, NCEP ATP III defines metabolic syndrome as the presence of any two of the following traits:

[0053] 1. Abdominal obesity defined as waist circumference > 102 cm (40 inches) in men and > 88 cm (35 inches) in women;

[0054] 2. Serum triglycerides > 150 mg / dL (1.7 mmol / L);

[0055] 3. Serum high-density lipoprotein (HDL) cholesterol < 40 mg / dL (1 mmol / L) in men and < 50 mg / dL (1.3 mmol / L) in women, or on medication for low HDL cholesterol; and

[0056] 4. Fasting plasma glucose > 100 mg / dL (5.6 mmol / L), or on medication for elevated glucose.

[0057] In one aspect, metabolic syndrome is identified in patients with three or more of the following risk factors defined by the American Heart Association, including high blood sugar, low levels of HDL cholesterol in the blood, high levels of triglycerides in the blood, large waist circumference, and high BP. In one aspect, a lipid profile or panel is a set of blood tests used to find abnormalities in lipids, such as cholesterol and triglycerides. Typically, the lab measures only three quantities, including total cholesterol, HDL, and triglycerides. LDL can be calculated from these three data. In one aspect, the lab requires patients to fast for 9 to 12 hours before screening. In one aspect, some diagnostic labs accept non-fasting samples.

[0058] In one aspect, the treatment methods provided herein are methods of achieving reduction in BP, enhancement of insulin sensitivity, increase in plasma non-esterified fatty acids (NEFA), reduction in plasma glucose levels, increase in urinary sodium excretion, arterial vasodilation, suppression of renin and aldosterone, reduction in apoptosis and hypertrophy, increase in cardiac diastolic compliance, induction of angiogenesis, increase in lipolysis, and browning of white adipocytes in a patient in need thereof.

[0059] MANP

[0060] MANP is an ANP analog that is a guanylate cyclase A (GC-A) / cyclic guanosine monophosphate (cGMP) activator. (McKie, Paul M, et al. Journal of the American College of Cardiology, 54(11): 1024-1032 (2009)). MANP is a 40 amino acid (AA) peptide that contains the 28 AA of native ANP with a unique 12 AA C-terminal extension. Compared to ANP, MANP is more resistant to degradation, enhances binding to the GC-A receptor, causes a greater and more sustained increase in sodium excretion, and causes more sustained suppression of aldosterone.

[0061] The methods provided herein can comprise, in part, treating a patient with a MANP. The disclosure provides polypeptides or nucleic acids encoding MANP polypeptides as shown in Table 1. As used herein, a “MANP” can have the amino acid sequence set forth in SEQ ID NO: 3, or can be a variant of the sequence set forth in SEQ ID NO: 3. As shown, a MANP is an ANP-based peptide whose amino acid sequence includes a 28-amino acid mature human ANP sequence (SLRRSSCFGGRMDRIGAQSGLGCNSFRY; SEQ ID NO: 1) and an additional 12-amino acid carboxy terminus (RITAREDKQGWA; SEQ ID NO: 2). The full-length sequence of a MANP is SLRRSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA (SEQ ID NO: 3). A representative nucleic acid sequence encoding a MANP is 5’-agcctgcggagatccagctgcttcgggggcaggatggacaggattggagcccagagcggactggg ctgtaacagcttccggtaccggataacagccagggaggacaagcagggctgggcctag-3’ (SEQ ID NO: 4). Figure 1

[0062] Table 1. Exemplary MANP polypeptides or nucleic acids encoding MANP polypeptides

[0063]

[0064] ​In an aspect, the MANP used in the methods provided herein can contain the entire amino acid sequence set forth in SEQ ID NO: 3. In an aspect, the MANP can contain the amino acid sequence set forth in SEQ ID NO: 3, wherein the amino acid sequence contains one or between one and ten (e.g., ten, between one and nine, between two and nine, between one and eight, between two and eight, between one and seven, between one and six, between one and five, between one and four, between one and three, two, or one) amino acid additions, subtractions, and / or substitutions. In an aspect, the MANP can contain the amino acid sequence set forth in SEQ ID NO: 3 with one, two, three, four, five, six, seven, eight, nine, or ten single amino acid residue additions, subtractions, or substitutions. In an aspect, the MANP can have one or more additions, subtractions, and / or substitutions within the C-terminal portion of SEQ ID NO: 3 (e.g., the last 12 amino acids of SEQ ID NO: 3). Examples of such polypeptides include, but are not limited to, polypeptides having the amino acid sequence set forth in SEQ ID NO: 3, wherein the threonine is deleted (SLRRSSCFGGRMDRIGAQSGLGCNSFRYRIAREDKQGWA; SEQ ID NO: 5), the tryptophan is substituted with tyrosine (SLRRSSCFGGRMDRIGAQSGLGCNSFRY RITAREDKQGYA; SEQ ID NO: 6), a serine is added between the lysine and glutamine (SLRRSSCFGGRMDRIGAQS GLGCNSFRYRITAREDKSQGWA; SEQ ID NO: 7), or any combination thereof. In an aspect, the MANP can lack the last three residues of SEQ ID NO: 3 (i.e., the glycine, tryptophan, and alanine residues of SEQ ID NO: 3, as set forth in SEQ ID NO: 8 (SLRRSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQ)). In an aspect, the MANP can have one or more additions, subtractions, and / or substitutions within the N-terminal portion of SEQ ID NO: 3 (e.g., the first six amino acids of SEQ ID NO: 3).Examples of such polypeptides include, but are not limited to, polypeptides having the amino acid sequence set forth in SEQ ID NO: 3, wherein four amino acids from urodilatin are added to the N-terminus (TAPRSLRRSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA; SEQ ID NO: 9), arginine residues at positions 3 and 4 are substituted with lysine residues (SLKKSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA; SEQ ID NO: 10), the D-isomer of serine is substituted at the sixth position (SLRRSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA; SEQ ID NO: 11), the D-isomer of arginine is substituted at the fourth position and the serine at the fifth position is deleted (SLRRSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA; SEQ ID NO: 12), a threonine residue substitutes the serine residues at positions 1, 5, and 6 (TLRRTTCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA; SEQ ID NO: 13), a tryptophan substitutes the leucine at position 2 (SWRRSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA; SEQ ID NO: 14), or any combination thereof.

[0065] In an aspect, any of the amino acid residues set forth in SEQ ID NO: 3 can be subtracted, and any amino acid residue (e.g., any of the 20 conventional amino acid residues or any other type of amino acid, such as ornithine or citrulline) can be added to the sequence set forth in SEQ ID NO: 3. In an aspect, the MANP can contain one or more chemical structures, such as epsilon-aminohexanoic acid; hydroxylated amino acids, such as 3-hydroxyproline, 4-hydroxyproline, (5R)-5-hydroxy-L-lysine, allohydroxylysine, and 5-hydroxy-L-norvaline; and / or glycosylated amino acids, such as amino acids containing monosaccharides (e.g., D-glucose, D-galactose, D-mannose, D-glucosamine, and D-galactosamine) or combinations of monosaccharides.

[0066] In one aspect, MANPs having one or more amino acid additions, subtractions, or substitutions relative to the representative MANP sequence set forth in SEQ ID NO: 3, also referred to herein as "variant" MANPs, can be produced using any suitable method. In one aspect, amino acid substitutions can be made by choosing substitutions that have little or no appreciable difference in their effect on maintaining (a) the structure of the peptide backbone in the region of the substitution, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the volume of the side chain. In one aspect, naturally occurring residues can be divided into groups based on common side-chain properties: (1) hydrophobic: norleucine, methionine, alanine, valine, leucine, and isoleucine; (2) neutral hydrophilic: cysteine, serine, and threonine; (3) acidic: aspartic acid and glutamic acid; (4) basic: asparagine, glutamine, histidine, lysine, and arginine; (5) residues that influence the overall charge of the molecule: glycine and proline; and (6) aromatic: tryptophan, tyrosine, and phenylalanine. Substitutions made within these groups can be considered conservative substitutions. Non-limiting examples of useful conservative substitutions can include, but are not limited to, valine to alanine, lysine to arginine, glutamine to asparagine, glutamic acid to aspartic acid, serine to cysteine, asparagine to glutamine, aspartic acid to glutamic acid, proline to glycine, arginine to histidine, leucine to isoleucine, isoleucine to leucine, arginine to lysine, leucine to methionine, leucine to phenylalanine, glycine to proline, threonine to serine, serine to threonine, tyrosine to tryptophan, phenylalanine to tyrosine, and / or leucine to valine.

[0067] Further examples of conservative substitutions that can be made at any position within a MANP useful in the methods described herein are listed in Table 2.

[0068] Table 2. Examples of conservative amino acid substitutions

[0069]

[0070]

[0071] In one aspect, a MANP can include one or more non-conservative substitutions. Non-conservative substitutions typically entail exchanging a member of one of the above- described classes for a member from another class. Such production can be desirable to provide a large number or alternative embodiments of such compounds. Whether an amino acid change results in a functional polypeptide can be readily determined by assaying the specific activity of the polypeptide variant, using, for example, a cGMP radioimmunoassay (RIA) or a cGMP ELISA assay.

[0072] In an aspect, a MANP can have a length of 35 to 45 amino acid residues (e.g., 35 to 40, 40 to 45, 35 to 37, 36 to 38, 37 to 39, 38 to 40, 39 to 41, 40 to 42, 41 to 43, 42 to 44, or 43 to 45 amino acid residues) in an aspect. In an aspect, a MANP can include an amino acid sequence as set forth in SEQ ID NO: 3, but with a particular number of amino acid substitutions. In an aspect, a MANP can have the amino acid sequence of SEQ ID NO: 3, but with one, two, three, four, or five amino acid substitutions. Examples of such amino acid sequences include, but are not limited to, a MANP with one or more L-amino acids replaced with a D-amino acid within the N-terminal region of the polypeptide (e.g., with a D-serine residue at position 6 as set forth in SEQ ID NO: 11, or with a D-arginine at position 4 as set forth in SEQ ID NO: 12).

[0073] In an aspect, a MANP can include an amino acid sequence having at least 90% (e.g., at least 90%, at least 92.5%, at least 95%, at least 97.5%, or 100%) sequence identity to the reference sequence set forth in SEQ ID NO: 3. The percent sequence identity is calculated by determining the number of aligned positions in the amino acid sequences, dividing the number of aligned positions by the total number of amino acids aligned, and then multiplying by 100. An aligned position refers to a position in the aligned amino acid sequences where the same amino acid occurs. Percent sequence identity can also be determined for any nucleic acid sequence.

[0074] The percent sequence identity between a particular nucleic acid or amino acid sequence and a sequence referenced by a particular sequence identification number is determined as follows. First, the nucleic acid or amino acid sequence is compared to the sequence set forth in the particular sequence identification number using the Bl2seq program from the independent version of BLASTZ which contains BLASTN version 2.0.14 and BLASTP version 2.0.14. This independent version of BLASTZ is available online at fr.com / blast or ncbi.nlm.nih.gov. Instructions for how to use the Bl2seq program can be found in the readme file accompanying BLASTZ. Bl2seq uses either the BLASTN or BLASTP algorithm for comparison. BLASTN is used for comparing nucleic acid sequences, while BLASTP is used for comparing amino acid sequences. For comparing two nucleic acid sequences, the options are set as follows: -i is set to the file containing the first nucleic acid sequence to be compared (e.g., C:\seq1.txt); -j is set to the file containing the second nucleic acid sequence to be compared (e.g., C:\seq2.txt); -p is set to blastn; -o is set to any desired file name (e.g., C:\output.txt); -q is set to -1; -r is set to 2; and all other options are left at their default settings. In one aspect, the following command can be used to generate an output file containing the comparison between the two sequences: C:\Bl2seq -i c:\seq1.txt -j c:\seq2.txt -p blastn -o c:\output.txt -q -1 -r 2. For comparing two amino acid sequences, the options for Bl2seq are set as follows: -i is set to the file containing the first amino acid sequence to be compared (e.g., C:\seq1.txt); -j is set to the file containing the second amino acid sequence to be compared (e.g., C:\seq2.txt); -p is set to blastp; -o is set to any desired file name (e.g., C:\output.txt); and all other options are left at their default settings. In one aspect, the following command can be used to generate an output file containing the comparison between the two amino acid sequences: C:\Bl2seq -i c:\seq1.txt -j c:\seq2.txt -p blastp -o c:\output.txt. If the two sequences being compared share homology, the designated output file will present those homologous regions as aligned sequences. If the two sequences being compared do not share homology, the designated output file will not present aligned sequences.

[0075] Once aligned, the number of matches is determined by counting the number of positions at which the same nucleotide or amino acid residue occurs in both sequences. The percent sequence identity is determined by dividing the number of matches by the length of the sequence set forth in the identified sequence (e.g., SEQ ID NO: 3) or by the length of the hinge (e.g., 20 contiguous nucleotides or amino acid residues from the sequence set forth in the identified sequence), and then multiplying the result by 100. In one aspect, an amino acid sequence having 37 matches when aligned with the sequence set forth in SEQ ID NO: 3 is 92.5% identical to the sequence set forth in SEQ ID NO: 3 (i.e., 37 ÷ 40 x 100 = 92.5). It should be noted that percent sequence identity values are rounded to the nearest tenth. In one aspect, 75.11, 75.12, 75.13, and 75.14 are rounded down to 75.1, while 75.15, 75.16, 75.17, 75.18, and 75.19 are rounded up to 75.2. It should also be noted that length values will always be whole numbers.

[0076] Isolated MANPs can be produced using any suitable method, including solid phase synthesis, and can be generated using manual techniques or automated techniques (e.g., using an Applied Biosystems (Foster City, CA) peptide synthesizer or a Biosearch Inc. (San Rafael, CA) automated peptide synthesizer). Disulfide bonds between cysteine residues can also be introduced by mild oxidation of linear polypeptides using KCN, as taught in U.S. Patent No. 4,757,048. MANPs can also be produced recombinantly.

[0077] Due to disulfide bonds between cysteine residues, the MANPs described herein are generally cyclic (see Figure 1 ). In one aspect, the thiol group on a cysteine residue can be replaced with an alternative group (e.g., -CH2CH2-). To replace the thiol group with a -CH2- group, in one aspect, the cysteine residue can be replaced with an a-aminobutyric acid. In one aspect, such cyclic analog polypeptides can be generated according to the method of Lebl and Hruby ((1984) Tetrahedron Lett., 25:2067-2068), or by employing the procedure disclosed in U.S. Patent No. 4,161,521.

[0078] In one aspect, ester bridges can be formed by reacting the OH of serine or threonine with the carboxyl group of aspartic acid or glutamic acid to produce a bridge having the structure -CH2CO2CH2-. Similarly, amides can be obtained by reacting the side chain of lysine with aspartic acid or glutamic acid to produce a bridge having the structure -CH2C(O)NH(CH)4-. Methods for synthesizing these bridges are known in the art (see, e.g., Schiller et al. (1985), Biochem. Biophys. Res. Comm., 127:558; and Schiller et al. (1985), Int. J. Peptide Protein Res., 25:171). In one aspect, when using Merrifield synthesis techniques, one method for preparing esters of the polypeptides of the application is to cleave the intact polypeptide from the resin under basic or acidic conditions (depending on the resin) in the presence of the desired alcohol. The C-terminus of the polypeptide can then be esterified directly upon release from the resin without the need to isolate the free acid. Amides of the polypeptides can also be prepared using techniques for converting carboxylic acid groups or precursors to amides, such as those known in the art. One method for forming amides at the C-terminal carboxyl group involves cleaving the polypeptide from a solid support with an appropriate amine, or cleaving in the presence of an alcohol to produce an ester, followed by aminolysis with the desired amine. Other bridging amino acid residues and reactions are provided in U.S. Patent No. 4,935,492. The preparation of peptide analogs that include nonpeptidyl linkages to connect amino acid residues is also known in the art.See, for example, Spatola et al. (1986), Life Sci., 38: 1243; Spatola (1983) Vega Data, 1(3); Morley (1980) Trends Pharm. Sci., 463-468; Hudson et al. (1979), Int. J. Pept. Prot. Res., 14: 177; Spatola, in Chemistry and Biochemistry of Amino Acid Peptides and Proteins, B. Weinstein, ed., Marcel Dekker, New York, p. 267 (1983); Hann (1982), J. Chem. Soc. Perkin Trans., 1: 307; Almquist et al. (1980), J. Med. Chem., 23: 1392; Jennings-White et al. (1982), Tetrahedron Lett., 23: 2533; EP 45665; Holladay et al. (1983), Tetrahedron Lett., 24: 4401; and Hruby (1982), Life Sci., 31: 189.

[0079] N-acyl derivatives of the amino groups of the polypeptides can be prepared by final condensation with N-acyl protected amino acids, or by acylation of protected or unprotected peptides. In one aspect, O-acyl derivatives can be prepared by acylation of free hydroxyl peptides or peptide resins. Either acylation can be performed using standard acylating reagents such as acyl halides, acid anhydrides, acylimidazoles, and the like. If desired, both N-acylation and O-acylation can be performed together.

[0080] In an aspect, the MANP can be pegylated, acetylated, or both. In an aspect, the polypeptide can be covalently attached to an oligomer, such as a short amphiphilic oligomer, which enables administration or improves pharmacokinetic or pharmacodynamic characteristics of the conjugated polypeptide. The oligomer can comprise a water-soluble polyethylene glycol (PEG) and / or a fat-soluble alkyl (short-, medium-, or long-chain fatty acid polymer, such as, but not limited to, palmitic acid, myristic acid, lauric acid, capric acid, or stearic acid). The fatty acid molecule can be attached to the free amino terminal end or any lysine side chain (epsilon amino group), and the lysine residue for such attachment can be located at the C- or N-terminus of the peptide. Linkage to PEG or another suitable polymer, or fusion to albumin or another suitable polypeptide, can result in a modified MANP having increased half-life compared to the unmodified MANP. Without being bound to a particular mechanism, the increased serum half-life can result from decreased proteolytic degradation, immune recognition, or cellular clearance of the modified MANP. Methods of modifying polypeptides by linkage to PEG (also known as “PEGylation”) or other polymers are known in the art and include those described in U.S. Patent No. 6,884,780; PCT Publication No. WO 2004 / 047871; Cataliotti et al. (2007), Trends Cardiovasc. Med., 17:10-14; Veronese and Mero (2008), BioDrugs, 22:315-329; Miller et al. (2006), Bioconjugate Chem., 17:267-274; and Veronese and Pasut (2005), Drug Discov. Today, 10:1451-1458, all of which are incorporated by reference herein in their entireties. Methods of modifying polypeptides by fusion to albumin are also known in the art and include those described in U.S. Patent Publication No. 2004 / 0086976 and Wang et al. (2004), Pharm. Res., 21:2105-2111, both of which are incorporated by reference herein in their entireties.

[0081] In an aspect, the MANP can be fused to the Fc domain of an immunoglobulin molecule (e.g., an IgG1 molecule) such that the fusion polypeptide is actively transported across an epithelial cell barrier via an Fc receptor. In an aspect, the polypeptide can be a cyclic polypeptide. Cyclic polypeptides can be obtained by incorporating cysteine residues; however, in an aspect, it is also contemplated to replace the sulfhydryl group on the cysteine residue with a -CH2-CH2- group. In an aspect, to replace the sulfhydryl group with a -CH2- group, the cysteine residue can be replaced with a similar alpha-aminobutyric acid. In an aspect, these cyclic analog peptides can be synthesized according to AboveMethods of Lebl and Hruby or by employing the procedure disclosed in U.S. Patent No. 4,161,521.

[0082] Salts of the carboxyl groups of MANP can be prepared by contacting the polypeptide with one or more equivalents of a desired base, such as, in an aspect, a metal hydroxide base (e.g., sodium hydroxide), a metal carbonate or bicarbonate base (e.g., sodium carbonate or sodium bicarbonate), or an amine base (e.g., triethylamine, triethanolamine, and the like). Acid addition salts of the polypeptide can be prepared by contacting the polypeptide with one or more equivalents of an inorganic or organic acid (e.g., hydrochloric acid).

[0083] The term "polypeptide" as used herein refers to a compound composed of two or more subunit amino acids, whether or not it is post-translationally modified (e.g., phosphorylated or glycosylated). The subunits can be connected by peptide bonds or other linkages, such as, in an aspect, ester or ether linkages. The term "amino acid" refers to natural and / or unnatural or synthetic amino acids, including D / L optical isomers.

[0084] The term "isolated" with respect to a polypeptide, as used herein, means that the polypeptide (1) is not associated with proteins found in nature, (2) is free of other proteins from the same source (e.g., free of human proteins), (3) is expressed by cells from a different species, or (4) does not exist in nature. In an aspect, an isolated polypeptide can be encoded by DNA or RNA, including synthetic DNA or RNA, or some combination thereof.

[0085] The term "substantially pure" as used herein with respect to a polypeptide means that the polypeptide is substantially free of other polypeptides, lipids, carbohydrates, and nucleic acids with which it is naturally associated. A substantially pure polypeptide can be any polypeptide removed from its natural environment and is at least 60% pure. A substantially pure polypeptide can be at least about 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% pure, or about 65% to 75%, 75% to 80%, 80% to 85%, 85% to 90%, 90% to 95%, or 95% to 99% pure. Typically, a substantially pure polypeptide will produce a single major band on a non-reducing polyacrylamide gel. In an aspect, a substantially pure polypeptide can be a chemically synthesized polypeptide.

[0086] Any method can be used to obtain a substantially pure polypeptide. In an aspect, common polypeptide purification techniques, such as affinity chromatography and HPLC, as well as polypeptide synthesis techniques can be used. Furthermore, any material can be used as a source material to obtain a substantially pure polypeptide. In an aspect, tissue from a wild-type or transgenic animal can be used as a source material. Furthermore, tissue culture cells engineered to overexpress a particular polypeptide can be used to obtain a substantially pure polypeptide. Furthermore, a polypeptide can be engineered to contain an amino acid sequence that allows the polypeptide to be captured onto an affinity matrix. In an aspect, tags such as c-myc, hemagglutinin, polyhistidine, or Flag TM tags (Kodak) can be used to aid in polypeptide purification. Such tags can be inserted anywhere within the polypeptide, including at the carboxy terminus or the amino terminus or in between. Other fusions that can be used include enzymes that aid in detection of the polypeptide, for example, alkaline phosphatase.

[0087] MANPs (e.g., variant MANPs having conservative and / or non-conservative substitutions relative to SEQ ID NO: 3), as well as fragments or variant MANPs of SEQ ID NO: 3 (e.g., fragments of any one of SEQ ID NOS: 3 to 14), can be screened for biological activity using any of a number of assays. In an aspect, activity can be assessed in vitro by testing the effect of a MANP on cGMP production in cultured cells (e.g., cultured cardiac fibroblasts, aortic endothelial cells, or glomerular cells). Cells can be exposed to a MANP (e.g., 10 -10 to 10 -4 M of the MANP), and the sample can be assayed to assess the effect of the polypeptide on cGMP production. In an aspect, cGMP production can be detected and measured using a competitive RIA cGMP kit (Perkin-Elmer, Boston, MA).

[0088] In an aspect, activity of a MANP can also be assessed in vivo by testing its effect on various factors, such as plasma cGMP levels, urinary cGMP excretion, renal net production of cGMP, glomerular filtration rate, BP, heart rate, hemodynamic function (such as cardiac output), pulmonary wedge pressure, systemic vascular resistance, and renal function (such as renal blood flow, urine volume, and sodium excretion rate) after administration to a patient (e.g., a human, non-human primate, rodent, dog, cat, pig, sheep, horse, or cow). In an aspect, such parameters can be assessed after hypertension is induced in the patient.

[0089] The term "nucleic acid" as used herein encompasses both RNA and DNA, including cDNA, genomic DNA, and synthetic (e.g., chemically synthesized) DNA. A nucleic acid can be double-stranded or single-stranded. In the case of single-stranded nucleic acids, the nucleic acid can be a sense strand or an antisense strand. Furthermore, a nucleic acid can be circular or linear.

[0090] As used herein, the term "isolated" with respect to a nucleic acid means a naturally occurring nucleic acid that is not immediately flanked by both of the sequences that immediately flank it in the naturally occurring genome of the organism from which it is derived, one at the 5' end and one at the 3' end. In an aspect, an isolated nucleic acid can be, but is not limited to, a recombinant DNA molecule of any length that is incorporated into a vector, into an autonomously replicating plasmid, or into a virus, or into a host cell genome, or is part of a hybrid or fusion nucleic acid sequence. In an aspect, an isolated nucleic acid can be, but is not limited to, a recombinant DNA molecule of any length that is not immediately flanked by both of the sequences that immediately flank it in the naturally occurring genome of the organism from which it is derived, one at the 5' end and one at the 3' end. Thus, an isolated nucleic acid includes a recombinant DNA that exists as a separate molecule independent of other sequences, as well as a recombinant DNA that is incorporated into a vector, into an autonomously replicating plasmid, or into a virus, or into a host cell genome, or is part of a hybrid or fusion nucleic acid sequence.

[0091] As used herein, the term "isolated" with respect to a nucleic acid also includes any non-naturally occurring nucleic acid, as a non-naturally occurring nucleic acid sequence does not exist in nature and does not have directly adjacent sequences in a naturally occurring genome. In an aspect, a non-naturally occurring nucleic acid, such as an engineered nucleic acid, is considered an isolated nucleic acid. Engineered nucleic acids can be made using common molecular cloning or chemical nucleic acid synthesis techniques. An isolated non-naturally occurring nucleic acid can exist independent of other sequences, or be incorporated into a vector, into an autonomously replicating plasmid, or into a virus, or into a host cell genome. In addition, a non-naturally occurring nucleic acid can include a nucleic acid molecule as part of a hybrid or fusion nucleic acid sequence.

[0092] It will be apparent to those skilled in the art that, in an aspect, nucleic acids that are present in hundreds to millions of other nucleic acid molecules in a cDNA or genomic library or gel slice containing a restriction digest of genomic DNA are not considered isolated nucleic acids.

[0093] In an aspect, an isolated nucleic acid molecule can be at least about 12 bases in length (e.g., at least about 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 40, 50, 60, 100, 120, 130, 140, 150, 250, 500, 750, 1000, 1500, 2000, 3000, 4000, or 5000 bases in length), and can hybridize under hybridizing conditions to a sense or antisense strand of a nucleic acid having a sequence encoding a MANP (e.g., a MANP having the sequence set forth in SEQ ID NO: 3, or a variant thereof). The hybridizing conditions can be moderately or highly stringent hybridization conditions.

[0094] As used herein, "moderately stringent hybridization conditions" means hybridization in about 42°C in a hybridization solution containing 25 mM KPO4 (pH 7.4), 5X SSC, 5X Denhart's solution, 50 μg / mL denatured, sonicated salmon sperm DNA, 50% formamide, 10% dextran sulfate, and 1-15 ng / mL probe (about 5 x 10 7 cpm / μg) with washes in about 50°C with wash solutions containing 2X SSC and 0.1% sodium dodecyl sulfate.

[0095] High stringency hybridization conditions means hybridization in about 42°C in a hybridization solution containing 25 mM KPO4 (pH 7.4), 5X SSC, 5X Denhart's solution, 50 μg / mL denatured, sonicated salmon sperm DNA, 50% formamide, 10% dextran sulfate, and 1-15 ng / mL probe (about 5 x 10 7 cpm / μg) with washes in about 65°C with wash solutions containing 0.2X SSC and 0.1% sodium dodecyl sulfate.

[0096] Isolated nucleic acid molecules encoding MANPs can be produced using standard techniques, including but not limited to common molecular cloning and chemical nucleic acid synthesis techniques. In an aspect, polymerase chain reaction (PCR) techniques can be used to obtain an isolated nucleic acid containing a nucleotide sequence encoding a MANP as provided herein. PCR refers to a procedure or technology in which a target nucleic acid is enzymatically amplified. Sequence information from the end of the region of interest or further away is typically employed to design oligonucleotide primers that are identical in sequence to opposite strands of the template to be amplified. PCR can be used to amplify specific sequences from DNA as well as RNA, including sequences from total genomic DNA or total cellular RNA. Primers are typically 14 to 40 nucleotides in length, but can range from 10 nucleotides to several hundred nucleotides in length. In an aspect, general PCR techniques are described in PCR Primer: A Laboratory Manual, edited by Dieffenbach and Dveksler, Cold Spring Harbor Laboratory Press, 1995. When RNA is used as the source of template, reverse transcriptase can be used to synthesize a complementary DNA (cDNA) strand. Ligase chain reaction, strand displacement amplification, self-sustained sequence replication, or nucleic acid sequence-based amplification can also be used to obtain an isolated nucleic acid. See, in an aspect, Lewis (1992), Genetic Engineering News, 12:1; Guatelli et al. (1990), Proc. Natl. Acad. Sci. USA, 87: 1874-1878; and Weiss (1991), Science, 254: 1292.

[0097] Isolated nucleic acids encoding MANPs can also be chemically synthesized, either as a single nucleic acid molecule (e.g., using phosphoramidite technology in the 3' to 5' direction using an automated DNA synthesizer) or as a series of oligonucleotides. In an aspect, one or more pairs of long oligonucleotides (e.g., > 100 nucleotides) containing the desired sequences can be synthesized, each pair containing a short segment of complementarity (e.g., about 15 nucleotides), such that when the oligonucleotide pairs are annealed, a duplex is formed. A DNA polymerase is used to extend the oligonucleotides, creating a single double-stranded nucleic acid molecule for each pair of oligonucleotides, which can then be ligated into a vector.

[0098] Furthermore, isolated nucleic acids encoding MANPs can be obtained by mutagenesis. In one aspect, reference sequences can be mutated using standard techniques, including oligonucleotide-directed mutagenesis and site-directed mutagenesis by PCR. See, Short Protocols in Molecular Biology, Chapter 8, Green Publishing Associates and John Wiley & Sons, Ausubel et al. eds., 1992. Non-limiting examples of variant MANPs are provided herein.

[0099] Vectors containing nucleic acids, such as those described herein, are also provided. A “vector” is a replicon, such as a plasmid, bacteriophage, or cosmid, into which another DNA segment can be inserted so as to bring about the replication of the inserted segment. An “expression vector” is a vector that includes one or more expression control sequences and an “expression control sequence” is a DNA sequence that controls and regulates the transcription and / or translation of another DNA sequence.

[0100] In an expression vector, a nucleic acid (e.g., a nucleic acid encoding a MANP) can be operably linked to one or more expression control sequences. As used herein, “operably linked” means incorporated into a gene construct in a manner that enables control of the expression of a coding sequence of interest by the expression control sequence. Examples of expression control sequences include promoters, enhancers, and transcription termination regions. A promoter is an expression control sequence that is comprised of a region of a DNA molecule, usually within 100 to 500 nucleotides upstream of the site where transcription starts (usually near the start site for RNA polymerase II). In order for a coding sequence to be under the control of a promoter, the translational start site of the translation reading frame of the polypeptide must be positioned between one and about 50 nucleotides downstream of the promoter. Enhancers provide expression specificity in terms of time, location, and level. Unlike promoters, enhancers can function when located at various distances from the transcriptional start site. Enhancers can also be located downstream of the transcriptional start site. A coding sequence is “operably linked” and “under the control” of an expression control sequence in a cell when RNA polymerase is able to transcribe the coding sequence into mRNA, which can then be translated into a protein encoded by the coding sequence. Thus, expression vectors can be used to produce antibodies as well as other multivalent molecules.

[0101] Suitable expression vectors include, but are not limited to, plasmids and viral vectors, which in one aspect are derived from bacteriophage, baculovirus, tobacco mosaic virus, herpes virus, cytomegalovirus, retrovirus, vaccinia virus, adenovirus, and adeno-associated virus. Many vectors and expression systems are commercially available from companies such as Novagen (Madison, WI), Clontech (Palo Alto, CA), Stratagene (La Jolla, CA), and Invitrogen / Life Technologies (Carlsbad, CA).

[0102] Expression vectors can include tag sequences designed to facilitate subsequent manipulation (e.g., purification or localization) of the expressed nucleic acid sequence. Tag sequences, such as green fluorescent protein (GFP), glutathione S-transferase (GST), polyhistidine, c-myc, hemagglutinin, or Flag TM Tag (Kodak, New Haven, CT) sequences are often expressed as fusions with the encoded polypeptide. Such tags can be inserted at any location within the polypeptide, including the carboxy terminus or the amino terminus.

[0103] Host cells containing the vectors are also provided. The term "host cell" is intended to include prokaryotic and eukaryotic cells, which can be introduced with a recombinant expression vector (e.g., a vector encoding a MANP). As used herein, "transformed" and "transfected" encompass introduction of a nucleic acid molecule (e.g., a vector) into a cell by any of a variety of techniques. While not limited to a particular technique, many such techniques are well-established in the art. In one aspect, suitable methods for transforming and transfecting host cells can be found in Sambrook et al., Molecular Cloning: A Laboratory Manual (2nd ed.), Cold Spring Harbor Laboratory, New York (1989). In one aspect, calcium phosphate precipitation, electroporation, heat shock, lipofection, microinjection, and viral-mediated transfer of nucleic acids can be used to introduce nucleic acids into cells. In addition, naked DNA can be delivered directly to cells in vivo as described elsewhere (U.S. Patent Nos. 5,580,859 and 5,589,466, both of which are incorporated by reference in their entireties). Host cells can express the encoded polypeptide, although it is noted that cells containing an isolated nucleic acid molecule provided herein need not express the polypeptide. Isolated nucleic acid molecules transformed into host cells can be integrated into the genome of the cell or maintained in an episomal state. Thus, host cells can be stably or transiently transfected with constructs containing an isolated nucleic acid molecule provided herein.

[0104] Any suitable method can be used to introduce the isolated nucleic acid molecule into a cell, either in vivo or in vitro. In one aspect, calcium phosphate precipitation, electroporation, heat shock, lipofection, microinjection, and viral-mediated nucleic acid transfer are methods that can be used to introduce the isolated nucleic acid molecule into a cell. In addition, naked DNA can be delivered directly to cells in vivo as described elsewhere (e.g., U.S. Patent Nos. 5,580,859 and 5,589,466, and continuations thereof). Furthermore, the isolated nucleic acid molecule can be introduced into a cell by generating a transgenic animal.

[0105] Any suitable method can be used to identify a cell that contains an isolated nucleic acid molecule encoding a MANP. Such methods include, but are not limited to, PCR and nucleic acid hybridization techniques, such as Northern and Southern analysis. In one aspect, immunohistochemical and biochemical techniques can be used to determine whether a cell contains a particular isolated nucleic acid molecule by detecting expression of a polypeptide encoded by the nucleic acid molecule.

[0106] The isolated MANP and nucleic acids encoding a MANP as described herein can be used to treat a patient identified as having a disorder such as hypertension with metabolic syndrome. Accordingly, one or more MANP or nucleic acids encoding one or more MANP can be incorporated into a composition for administration to a patient (e.g., a patient having hypertension, resistant hypertension (RH), and / or cardiorenal disease or at risk of developing hypertension, resistant hypertension, and / or cardiorenal disease). Any suitable method can be used to formulate and subsequently administer the composition. The dosage is generally dependent on the responsiveness of the patient to the administered agents, and the course of treatment lasts from several days to several months or longer, or until a suitable response is achieved. The optimal dosage, method of administration, and repetition rate are routinely determined by one of ordinary skill in the art. The optimal dosage can vary depending on the relative potency of each agent (e.g., MANP and Fs), and can generally be estimated based on EC 50 values found effective in in vitro and / or in vivo animal models. A composition containing one or more MANP as described herein can be given one or more times daily, weekly, monthly, or even less frequently, or can be administered continuously for a period of time (e.g., hours, days, or weeks). In one aspect, a MANP or a composition containing a MANP can be administered to a patient at a dose of at least about 0.01 ng MANP / kg to about 100 mg MANP / kg body weight. In one aspect, a MANP or a composition containing a MANP can be administered as an infusion for 1 to 30 days or longer (e.g., at a dose of about 1 pmol MANP / kg / min to about 500 nmol MANP / kg / min).

[0107] One or more MANPs or nucleic acids encoding one or more MANPs can be admixed, encapsulated, conjugated or otherwise associated with a mixture of other molecules, molecular structures or compounds, such as, in an aspect, liposomes, receptor or cell targeting molecules, or other formulations for oral, topical, or for aiding uptake, distribution and / or absorption.

[0108] In an aspect, the compositions of the present disclosure can comprise a MANP (or nucleic acid encoding a MANP) in combination with a pharmaceutically acceptable carrier. In an aspect, the pharmaceutically acceptable carrier includes a pharmaceutically acceptable solvent, suspending agent, or any other pharmacologically inert vehicle for delivering a polypeptide and / or other compound to a subject. The pharmaceutically acceptable carrier can be either liquid or solid, and can be selected with due regard to the intended mode of administration of a given pharmaceutical composition, so as to provide for the desired bulk, consistency, and other relevant transport and chemical properties when combined with the therapeutic compound(s) and any other components of the given pharmaceutical composition. Useful pharmaceutically acceptable carriers include, but are not limited to: water; saline solutions; binding agents (e.g., polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers (e.g., lactose or glucose and other sugars, gelatin, or calcium sulfate); lubricants (e.g., starch, polyethylene glycol, or sodium acetate); disintegrating agents (e.g., starch or sodium starch carbexyamate); and wetting agents (e.g., sodium lauryl sulfate).

[0109] Compositions and formulations for parenteral, intrathecal, or intraventricular administration include sterile aqueous solutions (e.g., sterile physiological saline), which can also contain buffers, diluents, and other suitable additives (e.g., penetration enhancers, carrier compounds, and other pharmaceutically acceptable carriers). In an aspect, compositions and formulations for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets, or tablets. Such compositions can further incorporate excipients, flavoring agents, diluents, emulsifiers, dispersion aids, or binders.

[0110] Pharmaceutical compositions include, but are not limited to, solutions, emulsions, aqueous suspensions, and liposome-containing formulations. These compositions can be generated from a wide variety of components that, in an aspect, include preformed liquids, self-emulsifying solids, and self-emulsifying semisolids. Emulsion formulations are particularly useful for oral delivery of therapeutic compositions due to their ease of formulation and efficacy of dissolution, absorption, and bioavailability. Liposomes can be particularly useful due to their specificity and the duration of action they provide from the perspective of drug delivery.

[0111] The compositions can contain any pharmaceutically acceptable salt, ester or salt of such ester, or any other compound, which upon administration to a subject is capable of providing (directly or indirectly) a biologically active metabolite or residue of the relevant compound (e.g., MANP). Accordingly, in an aspect, the present disclosure provides pharmaceutically acceptable salts of MANP, prodrugs, and pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. A prodrug is a therapeutic agent that is prepared in an inactive form and is converted to an active form (i.e., drug) within the body or within a cell thereof, by the action of endogenous enzymes or other chemicals and / or conditions. The term “pharmaceutically acceptable salt” refers to physiologically and pharmaceutically acceptable salts (i.e., salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects) of MANP that are useful in the methods provided herein. Examples of pharmaceutically acceptable salts include, but are not limited to, salts with cations such as sodium, potassium, calcium, or polyamines such as spermine; acid addition salts with inorganic acids such as hydrochloric, hydrobromic, sulfuric, phosphoric, or nitric acid; salts with organic acids such as acetic, citric, oxalic, palmitic, or fumaric acid; and salts with elemental anions such as bromine, iodine, or chlorine.

[0112] The compositions can also contain other auxiliary components conventionally found in pharmaceutical compositions. Thus, the compositions can also include compatible pharmaceutical active substances such as, in an aspect, antipruritics, astringents, local anesthetics, or anti-inflammatory agents, or additional substances that can be used in the physical formulation of the compositions in various dosage forms, such as dyes, flavorings, preservatives, antioxidants, opacifiers, thickening agents, and stabilizers. In addition, the compositions can be mixed with auxiliary agents, for example, lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorants, flavorings, penetration enhancers, and aromatic substances. However, when added, these materials should not unduly interfere with the biological activity of the other components of the composition.

[0113] In an aspect, the MANP can be formulated into a sustained release dosage form. In an aspect, the MANP can be formulated into a controlled release formulation. In an aspect, a coating, envelope, or protective matrix can be formulated to include one or more of the MANP described herein. Such coatings, envelopes, and protective matrices can be used to coat indwelling devices such as stents, catheters, and peritoneal dialysis tubes. In an aspect, the polypeptide can be incorporated into polymeric substances, liposomes, microemulsions, microparticles, nanoparticles, or waxes.

[0114] The pharmaceutical formulations disclosed herein, which can conveniently be presented in unit dosage form, can be prepared according to any suitable method known in the art of pharmacy, including conventional techniques. Such techniques include the step of bringing into association the active ingredient with the pharmaceutical carrier which constitutes one or more necessary ingredients. In general, the pharmaceutical formulations can be prepared by uniformly and intimately bringing into association the active ingredient with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, shaping the product. The pharmaceutical formulations can, if desired, be sterilized. Sterilization procedures can be accomplished using techniques known to those skilled in the art.

[0115] In an aspect, the MANP can be formulated for subcutaneous delivery via injection, depot polymer, drug patch, pump, or microparticle / nanoparticle. By way of example and not limitation, PCT Publication No. WO 2008 / 061355 discloses materials and methods for formulating polypeptides for delivery in a hydrogel tube. The polypeptides can be mixed with one or more excipients that are pharmaceutically acceptable and compatible with the polypeptides in the amounts used in the methods described herein. In an aspect, the polypeptides can be combined with one or more excipients such as, but not limited to, microcrystalline cellulose, colloidal silicon dioxide, lactose, starch, sorbitol, cyclodextrin, and combinations thereof. The excipients can be solid, semi-solid, or liquid materials that serve as vehicles, carriers, or media for the polypeptides. In an aspect, the polypeptides can be compressed, compacted, or extruded with one or more excipients prior to insertion into the hydrogel tube. Such formulations can result in pharmaceutical compositions with desired release characteristics, improved stability, and / or other desirable properties.

[0116] The pharmaceutical compositions can also include an adjuvant or excipient such as a glidant, dissolving agent, surfactant, diluent, binder, disintegrant, and / or lubricant. In an aspect, a dissolving agent can increase the rate of dissolution of the polypeptide from the dosage formulation, and in an aspect can include an organic acid and / or a salt of an organic acid (e.g., sodium citrate with citric acid). Other examples of excipients that can be used in such formulations include synthetic, semi-synthetic, modified, and natural polymers (e.g., lactose, dextrose, sucrose, trehalose, sorbitol, mannitol, starch, acacia, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, sugar syrup, PEG, cyclodextrin, alkoxylated modified cyclodextrin, hydroxyethyl cellulose, hydroxypropyl cellulose, microcrystalline cellulose, albumin, dextran, maltitol, xylitol, kaolin, and methyl cellulose). The polypeptides can also be mixed with a lubricant (e.g., talc, magnesium stearate, stearic acid, or mineral oil, calcium stearate, hydrogenated vegetable oil, sodium benzoate, sodium chloride, leucine polyethylene glycol, magnesium lauryl sulfate, or glyceryl monostearate), a wetting agent, an emulsor, and a suspending agent, or a preservative (e.g., methyl or propyl hydroxybenzoate).

[0117] Other agents that can be added to the pharmaceutical composition can alter the pH of the microenvironment upon dissolution of the MANP compound and establishment of a therapeutically effective plasma concentration profile. Such agents include inorganic acid salts and magnesium hydroxide. Other agents that can be used include surfactants and other solubilizing materials.

[0118] In an aspect, useful diluents include pharmaceutically acceptable inert fillers such as microcrystalline cellulose, lactose, sucrose, fructose, glucose, dextrose or other saccharides, calcium hydrogen phosphate, calcium sulfate, cellulose, ethyl cellulose, cellulose derivatives, kaolin, mannitol, lactitol, maltitol, xylitol, sorbitol or other sugar alcohols, dry starch, saccharides, dextrin, maltodextrin or other polysaccharides, inositol or combinations thereof. In an aspect, water-soluble diluents can be particularly useful.

[0119] Glidants can be used to improve the flow and compressibility of the composition ingredients during processing. In an aspect, useful glidants include colloidal silicon dioxide (also known as colloidal silicon dioxide, fumed silicon dioxide, light anhydrous silicic acid, silicic anhydride, and fumed silica).

[0120] Surfactants suitable for use in the pharmaceutical compositions described herein include, but are not limited to, sodium lauryl sulfate, polyethylene polyvinyl ester, polyethylene sorbitan fatty acid ester, polyoxyethylene castor oil derivative, polyoxyethylene alkyl ether, benzyl benzoate, cetrimide, cetyl alcohol, docusate sodium, glyceryl monooleate, glyceryl monostearate, glyceryl palmitostearate, lecithin, medium-chain triglyceride, monoethanolamine, oleic acid, poloxamer, polyvinyl alcohol, and sorbitan fatty acid ester.

[0121] In an aspect, suitable disintegrants include starch, sodium starch glycolate, crospovidone, crosscarmellose, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, pectin, potassium methacrylate-divinylbenzene copolymer, polyvinyl alcohol, acetamide, sodium bicarbonate, sodium carbonate, starch derivatives, dextrin, beta cyclodextrin, dextrin derivatives, magnesium oxide, clay, bentonite, and combinations thereof.

[0122] In an aspect, the MANP can be incorporated into a hydrogel delivery system. In an aspect, the MANP can be formulated for subcutaneous delivery to a patient via a xerogel-hydrogel system, which can release the polypeptide in a continuously sustained manner over an extended period of time. See, e.g., U.S. Patent No. 5,226,325 and PCT Publication No. WO 2004 / 071736, both of which are incorporated by reference in their entirety.

[0123] Liquid polymerizable materials useful for making hydrogel tubes include a variety of polymerizable, hydrophilic and ethylenically unsaturated compounds. See, for example, the compounds listed in PCT Publication No. WO 2008 / 061355. Mixtures of such hydrophilic monomers are typically used in the polymerization reaction. The type and ratio of monomers are selected to produce a polymer (e.g., a crosslinked homogenous polymer) that, when hydrated, has the desired properties (e.g., equilibrium water content (EWC) value and / or pore size) for the intended application or use.

[0124] In one aspect, polymerization of the hydrophilic monomer mixture can result in a homogenous hydrophilic copolymer that dissolves in aqueous media to varying degrees. In such cases, a small amount (e.g., up to about 3%) of a copolymerizable, multi-ethylenically unsaturated crosslinker can be included in the monomer mixture to obtain a water-insoluble as well as a water-swellable homogenous crosslinked copolymer. A homopolymer of lightly crosslinked methy lhe yl (hydroxy ethyl) acrylate (HEMA) has an EWC value of about 38%. Crosslinked copolymers of HEMA and N-(2-hydroxypropyl) methacrylamide (HPMA) have an EWC value lower than 38%, while crosslinked copolymers of HEMA and acrylamide exhibit an EWC value higher than 38 w / v%. Thus, depending on the useful or effective elution rate of the polypeptide, the copolymer hydrogel can be tailored to elute the polypeptide at the desired rate. Typically, the copolymer contains about 15 to about 70 weight percent of HEMA units and about 85 to 30 weight percent of a second ethylenic monomer, and thus has an EWC value in the range of about 20% to about 75%. In one aspect, the mixture of copolymers can further contain a small amount of a multi-ethylenically unsaturated crosslinker (e.g., ethylene glycol dimethacrylate ("EDMA") or trimethylolpropane trimethacrylate ("TMPTMA")).

[0125] In one aspect, a pharmaceutical composition for controlled release delivery of a MANP in a subject can include (a) a complex of a polypeptide (where the polypeptide has at least one basic functional group) and a polyanion derived from hexahydroxycyclohexane (where the polyanion has at least two negatively charged functional groups); and (b) a pharmaceutically acceptable carrier containing a biodegradable water-insoluble polymer. Such compositions are described, for example, in PCT Publication No. WO 2006 / 017852, and can be prepared in the form of a solution, suspension, dispersion, emulsion, drop, aerosol, cream, semi-solid, paste, capsule, tablet, solid implant, or microparticle. As used herein, the term "controlled release delivery" refers to the continuous delivery of an agent in vivo over a period of time (e.g., days to weeks or months) following administration. In one aspect, the sustained controlled release delivery of a MANP can be evidenced by the sustained therapeutic effect of the polypeptide over time (e.g., the sustained reduction of symptoms over time). The sustained delivery of the polypeptide can also be evidenced by detecting the presence of the polypeptide in vivo over time. The composition can provide a low initial burst, followed by a stable, controlled release of the polypeptide in vivo for an extended period of time (e.g., from days to months).

[0126] In one aspect, when the polypeptide and the polyanion are appropriately combined, a physically and chemically stable complex can be formed. The complex can take the form of a precipitate that is produced upon combining an aqueous preparation of the polypeptide and the polyanion. Optionally, one or more pharmaceutically acceptable excipients can be incorporated into the complex. Such excipients can be used as stabilizers for the polypeptide and / or the complex. Non-limiting examples of suitable excipients include sodium bisulfite, p-aminobenzoic acid, thiourea, glycine, methionine, mannitol, sucrose, and PEG.

[0127] The stable complex between the polypeptide and the polyanion can be incorporated into a pharmaceutically acceptable carrier containing a biodegradable water-insoluble polymer, optionally containing one or more excipients. The term "biodegradable water-insoluble polymer" refers to synthetic and natural polymers that are biocompatible and / or biodegradable for in vivo use. The term is also intended to include polymers that are insoluble or become insoluble in water or biological fluids at 37°C. The polymers can be purified using techniques known in the art (e.g., to remove monomers and oligomers). See, e.g., U.S. Patent No. 4,728,721. Useful polymers include, but are not limited to, polylactide, polyglycolide, poly(lactide-co-glycolide), polycaprolactone, polydioxanone, polycarbonates, polyhydroxybutyrate, polyalkylene oxalates, polyanhydrides, polyamides, polyesteramides, polyurethanes, polyacetals, polyorthocarbonates, polyphosphazenes, polyhydroxyvalerate, polyalkylene succinates, and polyorthoesters, as well as copolymers, block copolymers, branched copolymers, terpolymers, and combinations thereof.

[0128] The biodegradable water-insoluble polymer can also include a capped polymer, an uncapped polymer, or a mixture of capped and uncapped polymers. Capped polymers are generally defined as having capped carboxyl end groups, while uncapped polymers have free carboxyl end groups.

[0129] Factors to be considered when determining the appropriate molecular weight of the polymer can include the desired rate of polymer degradation, mechanical strength, and rate of polymer dissolution in a solvent. Useful molecular weights of the polymer can range from about 2,000 Daltons to about 150,000 Daltons, with a polydispersity of 1.1 to 2.8 in one aspect, depending on which polymer is selected for use.

[0130] The pharmaceutically acceptable carrier can be a carrier having environmentally responsive properties (e.g., thermosensitivity, pH sensitivity, or electrical sensitivity) in the form of an injectable solution or suspension, a granule, a film, a pellet, a cylinder, a disc, a microcapsule, a microsphere, a nanosphere, a microparticle, a wafer, a micelle, a liposome, or any other polymeric configuration useful for drug delivery.

[0131] Methods of forming various pharmaceutically acceptable polymeric carriers include those known in the art. See, e.g., U.S. Patent Nos. 6,410,044, 5,698,213, 6,312,679, 5,410,016, 5,529,914, 5,501,863, 4,938,763, 5,278,201, and 5,278,202; and PCT Publication No. WO 93 / 16687, all of which are incorporated by reference herein in their entireties.

[0132] When the polypeptide / polyanion complex is dispersed in a polymeric matrix to form a solid implant, which can be injected or implanted into a subject, a composition of the present disclosure can be produced. In one aspect, such implants can be prepared using conventional polymer melt processing techniques, such as extrusion, compression molding, and injection molding. Preparation of such implants can be performed under aseptic conditions, or alternatively, final sterilization by radiation (e.g., using gamma radiation or electron beam sterilization).

[0133] In one aspect, compositions in the form of microspheres can be prepared by encapsulating the polypeptide / polyanion complex in a polymeric carrier using various biocompatible and / or biodegradable polymers having properties suitable for delivery to different biological environments or to achieve specific functions. The rate of dissolution of the polypeptide, and thus the delivery of the polypeptide, is determined by factors such as the encapsulation technique, the polymer composition, the polymer cross-linking, the polymer thickness, the polymer solubility, and the size and solubility of the polypeptide / polyanion complex.

[0134] To prepare such microspheres, the polypeptide / polyanion complex to be encapsulated can be suspended in a polymer solution in an organic solvent, such that the polymer solution completely coats the polypeptide / polyanion complex. The suspension can then be subjected to microencapsulation techniques, such as spray drying, spray congealing, emulsification, or solvent evaporation emulsification. In one aspect, the suspended complex or microparticle can be transferred to a larger volume of an aqueous solution containing an emulsifier along with the polymer in an organic solvent, such that the organic solvent evaporates or diffuses away from the polymer, and the solidified polymer encapsulates the polypeptide / polyanion complex.

[0135] Emulsifiers that can be used to prepare the encapsulated polypeptide / polyanion complex include, for example, poloxamers and polyvinyl alcohol. Organic solvents that can be used in such methods include acetic acid, acetone, dichloromethane, ethyl acetate, chloroform, and other non-toxic solvents, depending on the nature of the polymer. The solvent is typically chosen to dissolve the polymer and be ultimately non-toxic.

[0136] In one aspect, the MANP can be formulated in a depot, which can provide a constant high exposure level and can rapidly reach a high exposure level (with a short or no lag phase). See, e.g., U.S. Pub. No. 2010 / 0266704, which is incorporated by reference herein in its entirety. The depot formulation can include a MANP or a pharmaceutically acceptable salt thereof (e.g., an acid addition salt with a mineral acid, a polymeric acid, or an organic acid). The acid addition salt can exist as a monovalent or divalent salt, depending on whether one or two acid equivalents are added.

[0137] As described in U.S. Pub. No. 2010 / 0266704, the depot formulation can contain two different linear poly(lactic-co-glycolic acid) (PLGA) polymers having a lactide:glycolide comonomer (L:G) molar ratio of 85: 15 to 65:35, wherein at least one polymer has a low intrinsic viscosity. Such formulations can provide sustained high plasma levels of the polypeptide over an extended period of time. Examples of suitable polymers include linear poly(D,L-lactide) and poly(D,L-lactide-co-glycolide) polymers sold under the trade names Resomer® and and by Boehringer Ingelheim Pharma GmBH & Co. KG (Ingelheim, Germany), Absorbable Polymers International (Pelham, AL), and Alkermes, Inc. (Cambridge, MA), respectively.

[0138] High exposure depot formulations for subcutaneous administration can exhibit immediate or at least very rapid onset of action, reaching therapeutic plasma concentrations within a short time period (e.g., one, two, three, four, five, six, or seven days after subcutaneous injection), and can exhibit sustained high exposure levels for about one month or more.

[0139] In one aspect, the depot formulations provided herein can contain two different PLGA polymers mixed or blended in a weight % ratio of 95:5 to 50:50 (e.g., 85:15 to 50:50, 80:20 to 60:40, 90:10, 85:15, 80:20, 75:25, 70:30, 65:35, 60:40, 55:45, or 50:50 wt%). In one aspect, the polymer with the higher intrinsic viscosity can have a higher weight % than the polymer with the lower intrinsic viscosity. In one aspect, the polymer with the higher intrinsic viscosity can have ester end groups. The depot formulation can contain additional polymers, including other linear or star-shaped PLGA polymers, or poly(D,L-lactide-co-glycolide) (PLG) or polylactic acid (PLA) polymers, provided that the favorable pharmacokinetic properties are maintained.

[0140] The polypeptide content (loading) of the depot formulation can be in the range of 1% to 30% (e.g., 10% to 25%, more preferably 15% to 20%). Loading is defined as the weight ratio of polypeptide to the total mass of the PLGA formulation.

[0141] The depot composition can be manufactured aseptically, or can be manufactured non- aseptically and terminally sterilized (e.g., using gamma irradiation). Terminal sterilization can result in a product with the highest assurance of sterility.

[0142] The depot composition can also contain one or more pharmaceutical excipients that modulate the release behavior of the polypeptide. Such excipients can be present in the composition in an amount of about 0.1% to about 50%. Suitable excipients include, but are not limited to, polyvinyl alcohol, polyvinylpyrrolidone, sodium carboxymethylcellulose, dextrin, PEG, surfactants such as poloxamers (also known as poly(oxyethylene-b- oxypropylene), poly(oxyethylene)-sorbitan-fatty acid esters, trade name Pluronic®), sorbitan fatty acid esters, lecithin, inorganic salts such as zinc carbonate, magnesium hydroxide, magnesium carbonate, protamine, and natural or synthetic polymers with amine residues such as polylysine.

[0143] ​The reservoir compositions can contain a mixture or blend of polymers that differ in composition, molecular weight, and / or polymer structure. A polymer blend is defined herein as a solid solution or suspension of two different linear polymers in one implant or microparticle. A reservoir mixture is defined herein as a mixture of two reservoir-like implants or microparticles or semi-solid formulations of different composition, with one or more PLGAs in each reservoir. Drug reservoir compositions in which two PLGAs are present as a polymer blend can be particularly useful.

[0144] The drug reservoir compositions can be in the form of implants, semi-solids (gels), liquid solutions, microparticles, or suspensions that solidify in situ once they are injected. The following paragraphs focus on polymeric microparticles, although the description also applies to implants, semi-solids, and liquids.

[0145] The microparticles can have diameters ranging from a few sub-microns to several millimeters (e.g., about 0.01 microns to about 2 mm, about 0.1 microns to about 500 microns, about 10 to about 200 microns, about 10 to about 130 microns, or about 10 to about 90 microns).

[0146] In one aspect, the microparticles can be mixed with or coated with an anti- agglomeration agent. Suitable anti-agglomeration agents include, for example, mannitol, dextrose, dextran, sucrose, sodium chloride, and water-soluble polymers such as polyvinyl alcohol, polyvinylpyrrolidone, and PEG.

[0147] The microparticles can be manufactured using methods known in the art, such as coacervation or phase separation, spray drying, or water-in-oil (W / O), water-in-oil-in-water (W / O / W), or water-in-oil-in-solid (S / O / W) emulsion / suspension methods followed by solvent extraction or solvent evaporation. Emulsion / suspension methods can be particularly useful and can include the following steps:

[0148] (i) preparing an internal organic phase comprising

[0149] (a) dissolving one or more polymers in a suitable organic solvent (e.g., ethyl acetate, acetone, THF, acetonitrile, or a halogenated hydrocarbon such as dichloromethane, chloroform, or hexafluoroisopropanol) or a mixture of solvents, and optionally dissolving / dispersing suitable additives; and

[0150] (b) dissolving / suspending / emulsifying a polypeptide in the polymer solution obtained in step (a);

[0151] (ii) preparing an external aqueous phase containing one or more stabilizing agents (e.g., poly(vinyl alcohol), hydroxyethylcellulose, hydroxypropylcellulose, poly(vinylpyrrolidone), or gelatin) and optionally a buffer salt;

[0152] (iii) mixing the internal organic phase with the external aqueous phase to form an emulsion; and

[0153] (iv) hardening the microparticles by solvent evaporation or solvent extraction, washing the microparticles (e.g., with water), collecting and drying the microparticles (e.g., by lyophilization or vacuum drying), and sieving the microparticles (e.g., by 140 μιη).

[0154] The dried microparticle composition can be terminally sterilized by gamma irradiation, either in bulk form or after dispensing into final containers. In one aspect, the bulk sterilized microparticles can be resuspended in a suitable vehicle and dispensed into suitable devices, such as dual chamber syringes, followed by lyophilization.

[0155] In one aspect, the microparticle depot composition can include a vehicle to facilitate reconstitution. Further, prior to administration, the microparticles can be suspended in a suitable injection vehicle (e.g., a water-based vehicle containing one or more pharmaceutical excipients, such as mannitol, sodium chloride, dextrose, dextrose, sucrose, or glycerol, and / or one or more non-ionic surfactants, such as poloxamer, poly(oxyethylene)-sorbitan-fatty acid ester, sodium carboxymethylcellulose, sorbitol, poly(vinylpyrrolidone), or aluminum monostearate).

[0156] Methods of administering MANPs

[0157] In one aspect, prior to administering a MANP or a nucleic acid encoding a MANP or one or more compositions comprising a MANP to a patient, an assessment is made to determine whether the patient is in need of treatment for hypertension with metabolic syndrome. In one aspect, prior to administering a MANP or a nucleic acid encoding a MANP to a patient, an assessment is made to determine whether the patient is in need of treatment for hypertension with metabolic syndrome. In one aspect, prior to administering a MANP or a nucleic acid encoding a MANP to a patient, an assessment is made to determine whether the patient is in need of treatment for hypertension with metabolic syndrome 0.5 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 1 week prior to administering a MANP or a nucleic acid encoding a MANP to a patient. In one aspect, in accordance with the present disclosure, the assessment includes one or more baseline measurements of a parameter selected from the group consisting of: plasma cGMP, BMI, waist circumference, serum triglycerides, HDL cholesterol, fasting plasma glucose, seated systolic BP, seated diastolic BP, plasma ANP-like peptide, insulin sensitivity, and plasma NEFA.

[0158] In an aspect, the patient in need thereof is a patient having a low baseline plasma cGMP measured prior to receiving the MANP according to the present disclosure. In an aspect, the low plasma baseline cGMP is less than 10 pmol / mL. In an aspect, the low plasma baseline cGMP is less than 8 pmol / mL. In an aspect, the low plasma baseline cGMP can be between 0.1 to 12 pmol / mL, 2 to 12 pmol / mL, 4 to 12 pmol / mL, 6 to 12 pmol / mL, 8 to 12 pmol / mL, or 10 to 12 pmol / mL. In an aspect, the low plasma baseline cGMP can be between 0.1 to 10 pmol / mL, 0.1 to 8 pmol / mL, 0.1 to 6 pmol / mL, 0.1 to 4 pmol / mL, or 0.1 to 2 pmol / mL. In an aspect, the low plasma baseline cGMP can be between 2 to 10 pmol / mL, 4 to 8 pmol / mL, 2 to 6 pmol / mL, 4 to 8 pmol / mL, or 6 to 10 pmol / mL. In an aspect, the plasma baseline plasma cGMP is measured with a commercial enzyme-linked immunosorbent assay (ELISA) kit.

[0159] In an aspect, the patient in need thereof is a patient having a BMI greater than 25 kg / m2. In an aspect, the patient in need thereof is a patient having a BMI greater than 30 kg / m2. In an aspect, the patient in need thereof is a patient having a BMI greater than 35 kg / m2. In an aspect, the patient in need thereof is a patient having a BMI between 20 to 50 kg / m2, 25 to 50 kg / m2, 30 to 50 kg / m2, 35 to 50 kg / m2, 40 to 50 kg / m2, or 45 to 50 kg / m2. In an aspect, the BMI can be between 20 to 45 kg / m2, 20 to 40 kg / m2, 20 to 35 kg / m2, or 20 to 30 kg / m2. In an aspect, the patient in need thereof is a patient having a BMI between 25 to 35 kg / m2, 30 to 40 kg / m2, or 35 to 45 kg / m2. 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 ​​​​​​​​​​​​​​​Patients with a BMI between [a certain threshold]. On one hand, BMI is measured by dividing a person's weight in kilograms (or pounds) by the square of their height in meters (or feet).

[0160] On one hand, patients requiring this service are male patients with a waist circumference greater than or equal to 102 cm. On another hand, patients requiring this service are male patients with a waist circumference between 90 and 300 cm, 90 and 250 cm, 90 and 200 cm, 90 and 150 cm, or 90 and 100 cm. On yet another hand, patients requiring this service are male patients with a waist circumference between 100 and 300 cm, 150 and 300 cm, 200 and 300 cm, or 250 and 300 cm. On yet another hand, patients requiring this service are male patients with a waist circumference between 100 and 250 cm, 150 and 200 cm, or 100 and 200 cm.

[0161] On one hand, patients requiring this service are women with a waist circumference greater than or equal to 88 cm. On the other hand, patients requiring this service are women with a waist circumference between 80 and 300 cm, 80 and 200 cm, 80 and 150 cm, or 80 and 100 cm. On the other hand, patients requiring this service are women with a waist circumference between 100 and 300 cm, 150 and 300 cm, 200 and 300 cm, or 250 and 300 cm. On the other hand, patients requiring this service are women with a waist circumference between 100 and 250 cm, 150 and 200 cm, or 100 and 200 cm.

[0162] In one aspect, the patient in need thereof is a patient having high serum triglycerides prior to receiving the MANP according to the present disclosure. In one aspect, high serum triglycerides is identified by serum triglycerides of 150 mg / dL or above. In one aspect, high serum triglycerides can be between 100 to 5000 mg / dL, 100 to 4000 mg / dL, 100 to 3000 mg / dL, 100 to 2000 mg / dL, 100 to 1000 mg / dL, 100 to 500 mg / dL, 100 to 300 mg / dL, or 100 to 200 mg / dL. In one aspect, high serum triglycerides can be between 200 to 5000 mg / dL, 300 to 5000 mg / dL, 500 to 5000 mg / dL, 1000 to 5000 mg / dL, 2000 to 5000 mg / dL, 3000 to 5000 mg / dL, or 4000 to 5000 mg / dL. In one aspect, high serum triglycerides can be between 200 to 4000 mg / dL, 300 to 3000 mg / dL, 500 to 2000 mg / dL, 1000 to 2000 mg / dL, 200 to 500 mg / dL, 300 to 1000 mg / dL, 500 to 2000 mg / dL, 1000 to 3000 mg / dL, or 2000 to 4000 mg / dL. In one aspect, serum triglycerides are measured by blood test after an overnight fast.

[0163] In one aspect, the patient in need thereof is a patient having low HDL cholesterol prior to receiving the MANP according to the present disclosure. In one aspect, low HDL cholesterol is identified by HDL cholesterol of less than 40 mg / dL in male patients. In one aspect, the patient in need thereof is a male patient having HDL cholesterol between 0.1 to 50 mg / dL, 0.1 to 40 mg / dL, 0.1 to 30 mg / dL, 0.1 to 20 mg / dL, or 0.1 to 10 mg / dL. In one aspect, the patient in need thereof is a male patient having HDL cholesterol between 10 to 50 mg / dL, 20 to 50 mg / dL, 30 to 50 mg / dL, or 40 to 50 mg / dL. In one aspect, the patient in need thereof is a male patient having HDL cholesterol between 10 to 40 mg / dL, 20 to 30 mg / dL, 10 to 30 mg / dL, or 20 to 40 mg / dL.

[0164] On one hand, low HDL cholesterol is identified by HDL cholesterol levels below 50 mg / dL in female patients. On the other hand, patients requiring this are those with HDL cholesterol levels between 0.1 and 60 mg / dL, 0.1 and 50 mg / dL, 0.1 and 40 mg / dL, 0.1 and 30 mg / dL, 0.1 and 20 mg / dL, 0.1 and 10 mg / dL, 0.1 and 5 mg / dL, 0.1 and 2.5 mg / dL, or 0.1 and 1 mg / dL. On the other hand, patients requiring this are those with HDL cholesterol levels between 10 and 60 mg / dL, 20 and 60 mg / dL, 30 and 60 mg / dL, 40 and 60 mg / dL, or 50 and 60 mg / dL. On the one hand, patients who require this are female patients with HDL cholesterol levels between 10 and 50 mg / dL, 20 and 40 mg / dL, 10 and 30 mg / dL, or 30 and 50 mg / dL.

[0165] On the one hand, patients who require this treatment are those receiving medication to increase their HDL cholesterol levels, which are typically low. On the other hand, HDL cholesterol is measured via blood tests.

[0166] In one aspect, the patient requiring this is one with high fasting plasma glucose as measured prior to receiving MANP according to this disclosure. In another aspect, high fasting plasma glucose is identified by a fasting plasma glucose level of 100 mg / dL or higher in the patient. In another aspect, the patient requiring this is one with fasting plasma glucose levels between 80 and 200 mg / dL, 80 and 180 mg / dL, 80 and 160 mg / dL, 80 and 140 mg / dL, 80 and 120 mg / dL, or 80 and 100 mg / dL. In yet another aspect, the patient requiring this is one with fasting plasma glucose levels between 100 and 200 mg / dL, 120 and 200 mg / dL, 140 and 200 mg / dL, 160 and 200 mg / dL, or 180 and 200 mg / dL. On one hand, patients requiring this measurement are those with fasting plasma glucose levels between 100 and 180 mg / dL, 120 and 160 mg / dL, or 100 and 140 mg / dL. On the other hand, patients requiring this measurement are those receiving medication to lower elevated blood glucose levels. Fasting plasma glucose is measured using Roche's glucose reagent (Indianapolis, IN) after an overnight fast.

[0167] In an aspect, the patient in need thereof presents with hypertension despite use of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten antihypertensive drugs. In an aspect, the patient in need thereof presents with hypertension despite use of one to ten, one to eight, one to six, one to four, one to two, two to ten, four to ten, six to ten, eight to ten, two to eight, four to six, one to three, two to four, three to five, six to eight, seven to nine, or nine to ten antihypertensive drugs. In an aspect, the antihypertensive drugs are selected from the group consisting of diuretics, angiotensin-converting enzyme (ACE) inhibitors, angiotensin 2 receptor blockers (ARBs), calcium channel blockers, beta blockers, renin inhibitors, alpha blockers, alpha-beta blockers, centrally acting agents, vasodilators, aldosterone antagonists, and combinations thereof.

[0168] In an aspect, the patient in need thereof does not present with any known hypersensitivity or allergic reaction to the MANP or components thereof, carperitide, other natriuretic peptides, or related compounds. In an aspect, the patient in need thereof is not pregnant. In an aspect, the patient in need thereof is not lactating. In an aspect, the patient in need thereof does not have a clinically significant history of endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery. In an aspect, the patient in need thereof does not have a history of severe allergies. In an aspect, the patient in need thereof is a patient who has not taken a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration of the MANP according to the present disclosure.

[0169] In an aspect, the patient in need thereof does not present with any trait selected from the group consisting of: a known hypersensitivity or allergic reaction to the MANP or components thereof, carperitide, other natriuretic peptides, or related compounds; is pregnant or lactating; has a clinically significant history of endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and has taken a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration.

[0170] In one aspect, the patient in need thereof (within 2 years prior to administration of the MANP according to the methods of the disclosure) has no history of alcohol abuse, illicit drug use, severe mental illness, a history of physical dependence on any opioid drug, or any history of drug abuse or addiction. In one aspect, the patient in need thereof has no history of blood donation difficulties, or has not donated blood or blood products within 45 days prior to administration. In one aspect, the patient in need thereof has no history of coronary artery disease or cerebrovascular disease or syncope. In one aspect, the patient in need thereof has no history of epilepsy or other seizure disorder. In one aspect, the patient in need thereof has no history of organ transplantation. In one aspect, the patient in need thereof has no malignancy within 5 years prior to administration of the MANP according to the methods of the disclosure. In one aspect, the malignancy refers to any kind of malignancy, including but not limited to bladder cancer, breast cancer, colon and rectum cancer, endometrial cancer, kidney cancer, leukemia, osteosarcoma, chordoma, glioblastoma, astrocytoma, skin cancer.

[0171] After a patient is identified as being in need of treatment for hypertension with metabolic syndrome, the patient can be treated according to the methods as described herein with the compositions as described herein. In one aspect, the composition comprising the MANP can be effectively administered to the patient in need thereof in any amount, at any frequency, and for any duration of time to achieve the desired result (e.g., to lower the BP of the patient, or to prevent or delay further elevation of the BP of the patient).

[0172] In one aspect, a method of treating hypertension with metabolic syndrome in a patient in need thereof having a low baseline plasma cGMP comprises increasing plasma cGMP by parenterally administering to the patient an effective dose of a MANP.

[0173] In one aspect, a method of treating hypertension with metabolic syndrome in a patient in need thereof comprises parenterally administering to the patient an effective dose of a MANP, the patient having a baseline plasma cGMP of less than 10 pmol / mL. In one aspect, a method of treating hypertension with metabolic syndrome in a patient in need thereof comprises parenterally administering to the patient an effective dose of a MANP, the patient having a baseline plasma cGMP of less than 8 pmol / mL.

[0174] In one aspect, the methods of the present disclosure increase plasma cGMP from baseline by 2 to 8 pmol / mL in a patient in need thereof within 0.5 hours after administration of an effective dose of MANP. In one aspect, the methods of the present disclosure increase plasma cGMP from baseline by between 0.1 to 10 pmol / mL, between 0.1 to 8 pmol / mL, between 0.1 to 6 pmol / mL, between 0.1 to 4 pmol / mL, or between 0.1 to 2 pmol / mL within 0.5 hours after administration of an effective dose of MANP. In one aspect, the methods of the present disclosure increase plasma cGMP from baseline by between 2 to 10 pmol / mL, between 4 to 10 pmol / mL, between 6 to 10 pmol / mL, or between 8 to 10 pmol / mL within 0.5 hours after administration of an effective dose of MANP. In one aspect, the methods of the present disclosure increase plasma cGMP from baseline by between 2 to 8 pmol / mL, between 4 to 6 pmol / mL, between 2 to 6 pmol / mL, or between 4 to 8 pmol / mL within 0.5 hours after administration of an effective dose of MANP.

[0175] In one aspect, the methods of the present disclosure increase plasma cGMP from baseline by 1 to 5 pmol / mL in a patient in need thereof within 4 hours after administration of an effective dose of MANP. In one aspect, the methods of the present disclosure increase plasma cGMP from baseline by between 0.1 to 6 pmol / mL, between 0.1 to 4 pmol / mL, or between 0.1 to 2 pmol / mL within 4 hours after administration of an effective dose of MANP. In one aspect, the methods of the present disclosure increase plasma cGMP from baseline by between 2 to 6 pmol / mL, between 4 to 6 pmol / mL, or between 2 to 4 pmol / mL within 4 hours after administration of an effective dose of MANP.

[0176] In one aspect, the methods of the present disclosure increase plasma cGMP in a patient in need thereof from baseline by 0.1 to 10 pmol / mL within 0.1 hour, 0.2 hour, 0.3 hour, 0.4 hour, 0.5 hour, 0.6 hour, 0.7 hour, 0.8 hour, 0.9 hour, 1 hour, 1.5 hour, 2 hour, 2.5 hour, 3 hour, 3.5 hour, 4 hour, 4.5 hour, 5 hour, 5.5 hour, 6 hour, 6.5 hour, 7 hour, 7.5 hour, 8 hour, 8.5 hour, 9 hour, 9.5 hour, 10 hour, 12 hour, or 24 hour after administration of an effective dose of MANP. In one aspect, the methods of the present disclosure increase plasma cGMP in a patient in need thereof from baseline by 0.1 to 10 pmol / mL between 0.1 and 6 hours, between 0.1 and 5 hours, between 0.1 and 4 hours, between 0.1 and 3 hours, between 0.1 and 2 hours, or between 0.1 and 1 hour after administration of an effective dose of MANP. In one aspect, the methods of the present disclosure increase plasma cGMP in a patient in need thereof from baseline by 0.1 to 10 pmol / mL between 1 and 6 hours, between 2 and 6 hours, or between 4 and 6 hours. The time after administration can be between 1 and 5 hours, between 2 and 4 hours, between 3 and 4 hours, between 0.5 and 2 hours, or between 1 and 3 hours after administration of an effective dose of MANP.

[0177] As used herein, “parenteral drug administration” refers to a drug that is administered by a route other than the digestive tract. The term parenteral is commonly used to refer to drugs that are administered by injection or infusion. In one aspect, the administration by parenteral administration is selected from the group consisting of subcutaneous administration (under the skin), intramuscular administration (in a muscle), intravenous administration (in a vein), and intrathecal administration (around the spinal cord).

[0178] In one aspect, a method of treating hypertension with metabolic syndrome in a patient in need thereof having a low baseline plasma cGMP comprises parenterally administering an effective amount of MANP at a dosage of about 0.01 ng polypeptide / kg to about 100 mg polypeptide / kg body weight (e.g., about 10 ng polypeptide / kg to about 50 mg polypeptide / kg, about 20 ng polypeptide / kg to about 10 mg polypeptide / kg, about 0.1 ng polypeptide / kg to about 20 ng polypeptide / kg, about 3 ng polypeptide / kg to about 10 ng polypeptide / kg, about 10 ng polypeptide / kg to about 50 ng polypeptide / kg, about 50 ng polypeptide / kg to about 100 μg polypeptide / kg, about 1 μg polypeptide / kg to about 10 μg polypeptide / kg, about 1 μg polypeptide / kg to about 5 μg polypeptide / kg, about 3 μg polypeptide / kg to about 7 μg polypeptide / kg, about 5 μg polypeptide / kg to about 10 μg polypeptide / kg, about 10 μg polypeptide / kg to about 20 μg polypeptide / kg, or about 20 μg polypeptide / kg to about 100 μg polypeptide / kg), although other dosages can provide beneficial results. In one aspect, the parenteral administration is subcutaneous administration. In one aspect, the composition comprising the MANP can be administered parenterally (e.g., by injection, such as subcutaneous injection, or by oral ingestion) at a dosage of about 0.5 ng polypeptide to about 20 g polypeptide (e.g., about 500 ng polypeptide to about 10 g polypeptide, about 1 μg polypeptide to about 2 g polypeptide, about 5 ng polypeptide to about 4 μg polypeptide, about 150 ng polypeptide to about 2 μg polypeptide, about 0.5 μg polypeptide to about 10 μg polypeptide, about 2.5 μg polypeptide to about 20 mg polypeptide, about 50 μg polypeptide to about 2 mg polypeptide, about 50 μg polypeptide to about 1 mg polypeptide, about 150 μg polypeptide to about 1.4 mg polypeptide, about 250 μg polypeptide to about 2 mg polypeptide, about 0.5 mg polypeptide to about 4 mg polypeptide, or about 1 mg polypeptide to about 20 mg polypeptide), although other dosages can provide beneficial results. In one aspect, the MANP can be administered by infusion at a dosage of, for example, about 1 pmol / kg / minute to about 500 nmol / kg / minute (e.g., about 1 to 10 pmol / kg / minute, about 10 to 100 pmol / kg / minute, about 100 to 300 pmol / kg / minute, about 250 to 500 pmol / kg / minute, about 500 pmol / kg / minute to about 1 nmol / kg / minute, about 1 to 10 nmol / kg / minute, about 10 to 100 nmol / kg / minute, about 100 to 300 nmol / kg / minute, or about 250 to 500 nmol / kg / minute).

[0179] In an aspect, a method of reducing BP in a patient in need thereof having low baseline plasma cGMP comprises increasing plasma cGMP compared to baseline by parenterally administering to the patient an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces seated systolic BP by between 0.1% and 15% compared to baseline seated systolic BP within 12 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces seated systolic BP by between 0.1% and 20%, between 0.1% and 15%, between 0.1% and 10%, between 0.1% and 5%, between 0.1% and 2.5%, or between 0.1% and 1% compared to baseline seated systolic BP within 12 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces seated systolic BP by between 5% and 20%, between 10% and 20%, or between 15% and 20% compared to baseline seated systolic BP within 12 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces seated systolic BP by between 5% and 15% or between 10% and 15% compared to baseline seated systolic BP within 12 hours after administration of an effective dose of MANP.

[0180] In an aspect, a method of reducing BP in a patient in need thereof reduces seated systolic BP from baseline seated systolic BP by between 0.1% and 20% within 0.1 hour, 0.2 hour, 0.3 hour, 0.4 hour, 0.5 hour, 0.6 hour, 0.7 hour, 0.8 hour, 0.9 hour, 1 hour, 1.5 hour, 2 hour, 2.5 hour, 3 hour, 3.5 hour, 4 hour, 4.5 hour, 5 hour, 5.5 hour, 6 hour, 6.5 hour, 7 hour, 7.5 hour, 8 hour, 8.5 hour, 9 hour, 9.5 hour, 10 hour, 12 hour, or 24 hour after administration of an effective dose of MANP. In an aspect, a method of the present disclosure reduces seated systolic BP from baseline seated systolic BP by between 0.1% and 20% in a patient in need thereof between 0.1 and 15 hours, between 0.1 and 12 hours, between 0.1 and 9 hours, between 0.1 and 6 hours, or between 0.1 and 3 hours after administration of an effective dose of MANP. In an aspect, a method of the present disclosure reduces seated systolic BP from baseline seated systolic BP by between 0.1% and 20% in a patient in need thereof between 3 and 15 hours, between 6 and 15 hours, between 9 and 15 hours, or between 12 and 15 hours after administration of an effective dose of MANP. In an aspect, a method of the present disclosure reduces seated systolic BP from baseline seated systolic BP by between 0.1% and 20% in a patient in need thereof between 3 and 12 hours, between 6 and 9 hours, between 3 and 9 hours, or between 6 and 12 hours after administration of an effective dose of MANP.

[0181] In an aspect, a method of reducing BP in a patient in need thereof reduces sitting systolic BP by 0.1 to 30 mmHg compared to baseline sitting systolic BP within 12 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces systolic BP by between 0.1 to 40 mmHg, between 0.1 to 30 mmHg, between 0.1 to 20 mmHg, between 0.1 to 10 mmHg, between 0.1 to 5 mmHg, between 0.1 to 2.5 mmHg, or between 0.1 to 1 mmHg compared to baseline sitting systolic BP within 12 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces systolic BP by between 10 to 40 mmHg, between 20 to 40 mmHg, or between 30 to 40 mmHg compared to baseline sitting systolic BP within 12 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces systolic BP by between 10 to 30 mmHg, between 10 to 20 mmHg, between 20 to 30 mmHg, or between 15 to 25 mmHg compared to baseline sitting systolic BP within 12 hours after administration of an effective dose of MANP.

[0182] In an aspect, a method of reducing BP in a patient in need thereof reduces the sitting systolic BP of the patient in need thereof from baseline sitting systolic BP by 0.1 to 40 mmHg within 0.1 hour, 0.2 hour, 0.3 hour, 0.4 hour, 0.5 hour, 0.6 hour, 0.7 hour, 0.8 hour, 0.9 hour, 1 hour, 1.5 hour, 2 hour, 2.5 hour, 3 hour, 3.5 hour, 4 hour, 4.5 hour, 5 hour, 5.5 hour, 6 hour, 6.5 hour, 7 hour, 7.5 hour, 8 hour, 8.5 hour, 9 hour, 9.5 hour, 10 hour, 12 hour, or 24 hour after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces the sitting systolic BP of the patient in need thereof from baseline sitting systolic BP by 0.1 to 40 mmHg between 0.1 and 15 hours, between 0.1 and 12 hours, between 0.1 and 9 hours, between 0.1 and 6 hours, or between 0.1 and 3 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces the sitting systolic BP of the patient in need thereof from baseline sitting systolic BP by 0.1 to 40 mmHg between 3 and 15 hours, between 6 and 15 hours, between 9 and 15 hours, or between 12 and 15 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces the sitting systolic BP of the patient in need thereof from baseline sitting systolic BP by 0.1 to 40 mmHg between 3 and 12 hours, between 6 and 9 hours, between 3 and 9 hours, or between 6 and 12 hours after administration of an effective dose of MANP.

[0183] In an aspect, a method of reducing BP in a patient in need thereof reduces the sitting diastolic BP of the patient in need thereof by 0.1% to 15% compared to baseline sitting diastolic BP within 6 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces the sitting diastolic BP of the patient in need thereof by 0.1% to 20%, between 0.1% and 15%, or between 0.1% and 10%, between 0.1% and 5%, between 0.1 and 2.5 mmHg, or between 0.1 and 1 mmHg compared to baseline sitting diastolic BP within 6 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces the sitting diastolic BP of the patient in need thereof by between 5% and 20%, between 10% and 20%, or between 15% and 20% within 6 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces the sitting diastolic BP of the patient in need thereof by between 5% and 15% or between 10% and 15% within 6 hours after administration of an effective dose of MANP.

[0184] In an aspect, a method of reducing BP in a patient in need thereof reduces seated diastolic BP from baseline seated diastolic BP by 0.1% to 20% within 0.1 hour, 0.2 hour, 0.3 hour, 0.4 hour, 0.5 hour, 0.6 hour, 0.7 hour, 0.8 hour, 0.9 hour, 1 hour, 1.5 hour, 2 hour, 2.5 hour, 3 hour, 3.5 hour, 4 hour, 4.5 hour, 5 hour, 5.5 hour, 6 hour, 6.5 hour, 7 hour, 7.5 hour, 8 hour, 8.5 hour, 9 hour, 9.5 hour, 10 hour, 12 hour, or 24 hour after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces seated diastolic BP from baseline seated diastolic BP by 0.1% to 20% between 0.1 and 8 hours, between 0.1 and 7 hours, between 0.1 and 6 hours, between 0.1 and 5 hours, between 0.1 and 4 hours, between 0.1 and 3 hours, between 0.1 and 2 hours, or between 0.1 and 1 hour after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces seated diastolic BP from baseline seated diastolic BP by 0.1% to 20% within a time after administration that is between 1 and 8 hours, between 2 and 8 hours, between 3 and 8 hours, between 4 and 8 hours, between 5 and 8 hours, between 6 and 8 hours, or between 7 and 8 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces seated diastolic BP from baseline seated diastolic BP by 0.1% to 20% within a time after administration that is between 2 and 6 hours or between 2 and 4 hours after administration of an effective dose of MANP.

[0185] In an aspect, a method of reducing BP in a patient in need thereof reduces seated diastolic BP by 0.1 to 30 mmHg compared to baseline seated diastolic BP within 6 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces diastolic BP by between 0.1 to 40 mmHg, between 0.1 to 30 mmHg, between 0.1 to 20 mmHg, between 0.1 to 10 mmHg, between 0.1 to 5 mmHg, between 0.1 to 2.5 mmHg, or between 0.1 to 1 mmHg compared to baseline seated diastolic BP within 6 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces diastolic BP by between 10 to 40 mmHg, between 20 to 40 mmHg, or between 30 to 40 mmHg compared to baseline seated diastolic BP within 6 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces diastolic BP by 10 to 30 mmHg compared to baseline seated diastolic BP within 6 hours after administration of an effective dose of MANP.

[0186] In an aspect, a method of reducing BP in a patient in need thereof reduces the sitting diastolic BP of the patient in need thereof from a baseline diastolic BP by 0.1 to 40 mmHg within 0.1 hour, 0.2 hour, 0.3 hour, 0.4 hour, 0.5 hour, 0.6 hour, 0.7 hour, 0.8 hour, 0.9 hour, 1 hour, 1.5 hour, 2 hour, 2.5 hour, 3 hour, 3.5 hour, 4 hour, 4.5 hour, 5 hour, 5.5 hour, 6 hour, 6.5 hour, 7 hour, 7.5 hour, 8 hour, 8.5 hour, 9 hour, 9.5 hour, 10 hour, 12 hour, or 24 hour after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces the sitting diastolic BP of the patient in need thereof from a baseline diastolic BP by 0.1 to 40 mmHg between 0.1 and 8 hours, between 0.1 and 7 hours, between 0.1 and 6 hours, between 0.1 and 5 hours, between 0.1 and 4 hours, between 0.1 and 3 hours, between 0.1 and 2 hours, or between 0.1 and 1 hour after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces the sitting diastolic BP of the patient in need thereof from a baseline diastolic BP by 0.1 to 40 mmHg between 1 and 8 hours, between 2 and 8 hours, between 3 and 8 hours, between 4 and 8 hours, between 5 and 8 hours, between 6 and 8 hours, or between 7 and 8 hours after administration of an effective dose of MANP. In an aspect, a method of reducing BP in a patient in need thereof reduces the sitting diastolic BP of the patient in need thereof from a baseline diastolic BP by 0.1 to 40 mmHg within 2 to 6 hours or within 2 to 4 hours after administration of an effective dose of MANP.

[0187] In one aspect, a method of reducing BP in a patient in need thereof having low baseline plasma cGMP comprises parenterally administering an effective dose of MANP at a dose of about 0.01 ng polypeptide / kg to about 100 mg polypeptide / kg body weight (e.g., about 10 ng polypeptide / kg to about 50 mg polypeptide / kg, about 20 ng polypeptide / kg to about 10 mg polypeptide / kg, about 0.1 ng polypeptide / kg to about 20 ng polypeptide / kg, about 3 ng polypeptide / kg to about 10 ng polypeptide / kg, about 10 ng polypeptide / kg to about 50 ng polypeptide / kg, about 50 ng polypeptide / kg to about 100 μg polypeptide / kg, about 1 μg polypeptide / kg to about 10 μg polypeptide / kg, about 1 μg polypeptide / kg to about 5 μg polypeptide / kg, about 3 μg polypeptide / kg to about 7 μg polypeptide / kg, about 5 μg polypeptide / kg to about 10 μg polypeptide / kg, about 10 μg polypeptide / kg to about 20 μg polypeptide / kg, or about 20 μg polypeptide / kg to about 100 μg polypeptide / kg), although other doses can provide beneficial results. In one aspect, a composition comprising MANP can be administered parenterally (e.g., by injection, such as subcutaneous injection, or by oral ingestion) at a dose of about 0.5 ng polypeptide to about 20 g polypeptide (e.g., about 500 ng polypeptide to about 10 g polypeptide, about 1 μg polypeptide to about 2 g polypeptide, about 5 ng polypeptide to about 4 μg polypeptide, about 150 ng polypeptide to about 2 μg polypeptide, about 0.5 μg polypeptide to about 10 μg polypeptide, about 2.5 μg polypeptide to about 20 mg polypeptide, about 50 μg polypeptide to about 2 mg polypeptide, about 50 μg polypeptide to about 1 mg polypeptide, about 150 μg polypeptide to about 1.4 mg polypeptide, about 250 μg polypeptide to about 2 mg polypeptide, about 0.5 mg polypeptide to about 4 mg polypeptide, or about 1 mg polypeptide to about 20 mg polypeptide), although other doses can provide beneficial results. In one aspect, MANP can be administered by infusion at a dose of, e.g., about 1 pmol / kg / minute to about 500 nmol / kg / minute (e.g., about 1 to 10 pmol / kg / minute, about 10 to 100 pmol / kg / minute, about 100 to 300 pmol / kg / minute, about 250 to 500 pmol / kg / minute, about 500 pmol / kg / minute to about 1 nmol / kg / minute, about 1 to 10 nmol / kg / minute, about 10 to 100 nmol / kg / minute, about 100 to 300 nmol / kg / minute, or about 250 to 500 nmol / kg / minute).

[0188] In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof having a low baseline plasma cGMP comprises increasing plasma cGMP compared to baseline by parenterally administering to the patient an effective dose of MANP. In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide compared to baseline plasma ANP-like peptide by between 1 and 70 pg / mL within 0.5 hours after administration of an effective dose of MANP. In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide compared to baseline plasma ANP-like peptide by between 0.1 and 100 pg / mL, between 0.1 and 90 pg / mL, between 0.1 and 80 pg / mL, between 0.1 and 70 pg / mL, between 0.1 and 60 pg / mL, between 0.1 and 50 pg / mL, between 0.1 and 40 pg / mL, between 0.1 and 30 pg / mL, between 0.1 and 20 pg / mL, between 0.1 and 10 pg / mL, between 0.1 and 5 pg / mL, between 0.1 and 2.5 pg / mL, or between 0.1 and 1 pg / mL within 0.5 hours after administration of an effective dose of MANP. In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide compared to baseline plasma ANP-like peptide by between 10 and 100 pg / mL, between 20 and 100 pg / mL, between 30 and 100 pg / mL, between 40 and 100 pg / mL, between 50 and 100 pg / mL, between 60 and 100 pg / mL, between 70 and 100 pg / mL, between 80 and 100 pg / mL, or between 90 and 100 pg / mL within 0.5 hours after administration of an effective dose of MANP. In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide compared to baseline plasma ANP-like peptide by between 10 and 90 pg / mL, between 20 and 80 pg / mL, between 30 and 70 pg / mL, between 40 and 60 pg / mL, between 10 and 30 pg / mL, between 20 and 40 pg / mL, between 30 and 50 pg / mL, between 40 and 60 pg / mL, between 50 and 70 pg / mL, between 60 and 80 pg / mL, or between 70 and 90 pg / mL within 0.5 hours after administration of an effective dose of MANP.

[0189] In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide from baseline plasma ANP-like peptide by 0.1 to 100 pg / mL within 0.1 hour, 0.2 hour, 0.3 hour, 0.4 hour, 0.5 hour, 0.6 hour, 0.7 hour, 0.8 hour, 0.9 hour, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, 4.5 hours, 5 hours, 5.5 hours, 6 hours, 6.5 hours, 7 hours, 7.5 hours, 8 hours, 8.5 hours, 9 hours, 9.5 hours, 10 hours, 12 hours, or 24 hours after administration of an effective dose of MANP. In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide from baseline plasma ANP-like peptide by 0.1 to 100 pg / mL between 0.1 and 2 hours, between 0.1 and 1.5 hours, or between 0.1 and 1 hour after administration of an effective dose of MANP. In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide from baseline plasma ANP-like peptide by 0.1 to 100 pg / mL between 0.5 and 2 hours, between 1 and 2 hours, or between 1.5 and 2 hours after administration of an effective dose of MANP. In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide from baseline plasma ANP-like peptide by 0.1 to 100 pg / mL between 0.5 and 1.5 hours after administration of an effective dose of MANP.

[0190] In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide by 1 to 40 pg / mL compared to baseline plasma ANP-like peptide within 1 hour after administration of an effective dose of MANP. In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide by between 0.1 to 60 pg / mL, between 0.1 to 50 pg / mL, between 0.1 to 40 pg / mL, between 0.1 to 30 pg / mL, between 0.1 to 20 pg / mL, or between 0.1 to 10 pg / mL compared to baseline plasma ANP-like peptide within 1 hour after administration of an effective dose of MANP. In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide by can be between 10 to 60 pg / mL, between 20 to 60 pg / mL, between 30 to 60 pg / mL, between 40 to 60 pg / mL, between 50 to 60 pg / mL, between 10 to 30 pg / mL, between 20 to 40 pg / mL, or between 30 to 50 pg / mL compared to baseline plasma ANP-like peptide within 1 hour after administration of an effective dose of MANP.

[0191] In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide by 0.1 to 60 pg / mL compared to baseline plasma ANP-like peptide within 0.1 hour, 0.2 hour, 0.3 hour, 0.4 hour, 0.5 hour, 0.6 hour, 0.7 hour, 0.8 hour, 0.9 hour, 1 hour, 1.5 hour, 2 hour, 2.5 hour, 3 hour, 3.5 hour, 4 hour, 4.5 hour, 5 hour, 5.5 hour, 6 hour, 6.5 hour, 7 hour, 7.5 hour, 8 hour, 8.5 hour, 9 hour, 9.5 hour, 10 hour, 12 hour, or 24 hour after administration of an effective dose of MANP. In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide by 0.1 to 60 pg / mL compared to baseline plasma ANP-like peptide between 0.1 to 2 hour, between 0.1 to 1.5 hour, or between 0.1 to 1 hour after administration of an effective dose of MANP. In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide by 0.1 to 60 pg / mL compared to baseline plasma ANP-like peptide between 0.5 to 2 hour, between 1 to 2 hour, or between 1.5 to 2 hour after administration of an effective dose of MANP. In an aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof increases plasma ANP-like peptide by 0.1 to 60 pg / mL compared to baseline plasma ANP-like peptide between 0.5 to 1.5 hour after administration of an effective dose of MANP.

[0192] In an aspect, the level of plasma ANP-like peptide is measured using an established ANP radioimmunoassay (RIA) (Mayo Clinic (Rochester, MN) and Phoenix Pharmaceuticals, Mountain View, CA).

[0193] In one aspect, a method of increasing plasma ANP-like peptide in a patient in need thereof having a baseline plasma cGMP of less than 10 pmol / mL includes parenterally administering an effective dose of MANP at a dose of about 0.01 ng polypeptide / kg to about 100 mg polypeptide / kg body weight (e.g., about 10 ng polypeptide / kg to about 50 mg polypeptide / kg, about 20 ng polypeptide / kg to about 10 mg polypeptide / kg, about 0.1 ng polypeptide / kg to about 20 ng polypeptide / kg, about 3 ng polypeptide / kg to about 10 ng polypeptide / kg, about 10 ng polypeptide / kg to about 50 ng polypeptide / kg, about 50 ng polypeptide / kg to about 100 μg polypeptide / kg, about 1 μg polypeptide / kg to about 10 μg polypeptide / kg, about 1 μg polypeptide / kg to about 5 μg polypeptide / kg, about 3 μg polypeptide / kg to about 7 μg polypeptide / kg, about 5 μg polypeptide / kg to about 10 μg polypeptide / kg, about 10 μg polypeptide / kg to about 20 μg polypeptide / kg, or about 20 μg polypeptide / kg to about 100 μg polypeptide / kg), although other doses can provide beneficial results. In one aspect, a composition comprising MANP can be administered parenterally (e.g., by injection, such as subcutaneous injection, or by oral ingestion) at a dose of about 0.5 ng polypeptide to about 20 g polypeptide (e.g., about 500 ng polypeptide to about 10 g polypeptide, about 1 μg polypeptide to about 2 g polypeptide, about 5 ng polypeptide to about 4 μg polypeptide, about 150 ng polypeptide to about 2 μg polypeptide, about 0.5 μg polypeptide to about 10 μg polypeptide, about 2.5 μg polypeptide to about 20 mg polypeptide, about 50 μg polypeptide to about 2 mg polypeptide, about 50 μg polypeptide to about 1 mg polypeptide, about 150 μg polypeptide to about 1.4 mg polypeptide, about 250 μg polypeptide to about 2 mg polypeptide, about 0.5 mg polypeptide to about 4 mg polypeptide, or about 1 mg polypeptide to about 20 mg polypeptide), although other doses can provide beneficial results. In one aspect, MANP can be administered by infusion at a dose of, for example, about 1 pmol / kg / minute to about 500 nmol / kg / minute (e.g., about 1 to 10 pmol / kg / minute, about 10 to 100 pmol / kg / minute, about 100 to 300 pmol / kg / minute, about 250 to 500 pmol / kg / minute, about 500 pmol / kg / minute to about 1 nmol / kg / minute, about 1 to 10 nmol / kg / minute, about 10 to 100 nmol / kg / minute, about 100 to 300 nmol / kg / minute, or about 250 to 500 nmol / kg / minute).

[0194] In one aspect, a method for lowering plasma glucose in a patient requiring this includes administering an effective dose of MANP parenterally to the patient. In another aspect, a method for lowering plasma glucose levels in a patient requiring this includes reducing plasma glucose by 1 to 17 mg / mL compared to baseline plasma glucose within 4 hours following administration of an effective dose of MANP. In another aspect, a method for lowering plasma glucose levels in a patient requiring this includes reducing plasma glucose levels by 0.1 to 20 mg / mL, 0.1 to 15 mg / mL, 0.1 to 10 mg / mL, 0.1 to 5 mg / mL, or 0.1 to 1 mg / mL compared to baseline plasma glucose within 4 hours following administration of an effective dose of MANP. In yet another aspect, a method for lowering plasma glucose levels in a patient requiring this includes reducing plasma glucose levels by 5 to 20 mg / mL, 10 to 20 mg / mL, or 15 to 20 mg / mL compared to baseline plasma glucose within 4 hours following administration of an effective dose of MANP. On the one hand, one method for reducing plasma glucose levels in patients who require this is to reduce plasma glucose levels by 5 to 15 mg / mL, 5 to 10 mg / mL, 10 to 15 mg / mL, or 7.5 to 12.5 mg / mL compared to baseline plasma glucose levels within 4 hours after administration of an effective dose of MANP.

[0195] On the one hand, one method for reducing plasma glucose levels in patients with this need is to reduce plasma glucose levels by 0.1 to 20 mg / mL compared to baseline plasma glucose within 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 12, or 24 hours after administration of an effective dose of MANP. In one aspect, a method for reducing the plasma glucose level of a patient in need of this treatment reduces the plasma glucose level by 0.1 to 20 mg / mL compared to baseline plasma glucose within 0.1 to 6 hours, 0.1 to 5 hours, 0.1 to 4 hours, 0.1 to 3 hours, 0.1 to 2 hours, or 0.1 to 1 hour after administration of an effective dose of MANP. In another aspect, a method for reducing the plasma glucose level of a patient in need of this treatment reduces the plasma glucose level by 0.1 to 20 mg / mL compared to baseline plasma glucose within 1 to 6 hours, 2 to 6 hours, 3 to 6 hours, 4 to 6 hours, 5 to 6 hours, 2 to 4 hours, or 3 to 5 hours after administration of an effective dose of MANP.

[0196] In an aspect, plasma glucose is measured using a Roche glucose reagent (Indianapolis, IN) after an overnight fast.

[0197] In an aspect, a method of reducing plasma glucose in a patient in need thereof comprises parenterally administering an effective dose of MANP at a dose of about 0.01 ng polypeptide / kg to about 100 mg polypeptide / kg body weight (e.g., about 10 ng polypeptide / kg to about 50 mg polypeptide / kg, about 20 ng polypeptide / kg to about 10 mg polypeptide / kg, about 0.1 ng polypeptide / kg to about 20 ng polypeptide / kg, about 3 ng polypeptide / kg to about 10 ng polypeptide / kg, about 10 ng polypeptide / kg to about 50 ng polypeptide / kg, about 50 ng polypeptide / kg to about 100 μg polypeptide / kg, about 1 μg polypeptide / kg to about 10 μg polypeptide / kg, about 1 μg polypeptide / kg to about 5 μg polypeptide / kg, about 3 μg polypeptide / kg to about 7 μg polypeptide / kg, about 5 μg polypeptide / kg to about 10 μg polypeptide / kg, about 10 μg polypeptide / kg to about 20 μg polypeptide / kg, or about 20 μg polypeptide / kg to about 100 μg polypeptide / kg), although other doses can provide beneficial results. In an aspect, a composition comprising MANP can be administered parenterally (e.g., by injection, such as subcutaneous injection, or by oral ingestion) at a dose of about 0.5 ng polypeptide to about 20 g polypeptide (e.g., about 500 ng polypeptide to about 10 g polypeptide, about 1 μg polypeptide to about 2 g polypeptide, about 5 ng polypeptide to about 4 μg polypeptide, about 150 ng polypeptide to about 2 μg polypeptide, about 0.5 μg polypeptide to about 10 μg polypeptide, about 2.5 μg polypeptide to about 20 mg polypeptide, about 50 μg polypeptide to about 2 mg polypeptide, about 50 μg polypeptide to about 1 mg polypeptide, about 150 μg polypeptide to about 1.4 mg polypeptide, about 250 μg polypeptide to about 2 mg polypeptide, about 0.5 mg polypeptide to about 4 mg polypeptide, or about 1 mg polypeptide to about 20 mg polypeptide), although other doses can provide beneficial results. In an aspect, MANP can be administered by infusion at a dose of, e.g., about 1 pmol / kg / minute to about 500 nmol / kg / minute (e.g., about 1 to 10 pmol / kg / minute, about 10 to 100 pmol / kg / minute, about 100 to 300 pmol / kg / minute, about 250 to 500 pmol / kg / minute, about 500 pmol / kg / minute to about 1 nmol / kg / minute, about 1 to 10 nmol / kg / minute, about 10 to 100 nmol / kg / minute, about 100 to 300 nmol / kg / minute, or about 250 to 500 nmol / kg / minute).

[0198] In an aspect, a method of increasing insulin sensitivity in a patient in need thereof comprises parenterally administering to the patient an effective dose of MANP. In an aspect, a method of increasing insulin sensitivity in a patient in need thereof increases the patient's HOMA2-insulin sensitivity (HOMA2-S) value by 0.1% to 35% compared to the baseline HOMA2-S value within 4 hours after administration of the effective dose of MANP. In an aspect, a method of increasing insulin sensitivity in a patient in need thereof increases the patient's HOMA2-S value by between 0.1% to 40%, between 0.1% to 30%, between 0.1% to 20%, between 0.1% to 10%, between 0.1% to 5%, between 0.1% to 2.5%, or between 0.1% to 1% compared to the baseline HOMA2-S value within 4 hours after administration of the effective dose of MANP. In an aspect, a method of increasing insulin sensitivity in a patient in need thereof increases the patient's HOMA2-S value by between 10% to 40%, between 20% to 40%, or between 30% to 40% compared to the baseline HOMA2-S value within 4 hours after administration of the effective dose of MANP. In an aspect, a method of increasing insulin sensitivity in a patient in need thereof increases the patient's HOMA2-S value by between 10% to 30%, between 10% to 20%, between 20% to 30%, or between 15% to 25% compared to the baseline HOMA2-S value within 4 hours after administration of the effective dose of MANP.

[0199] In an aspect, a method of increasing insulin sensitivity in a patient in need thereof increases the patient's HOMA2-S value by between 0.1% and 40% compared to the baseline HOMA2-S value within 0.1 hour, 0.2 hour, 0.3 hour, 0.4 hour, 0.5 hour, 0.6 hour, 0.7 hour, 0.8 hour, 0.9 hour, 1 hour, 1.5 hour, 2 hour, 2.5 hour, 3 hour, 3.5 hour, 4 hour, 4.5 hour, 5 hour, 5.5 hour, 6 hour, 6.5 hour, 7 hour, 7.5 hour, 8 hour, 8.5 hour, 9 hour, 9.5 hour, 10 hour, 12 hour, or 24 hour after administration of an effective dose of MANP. In an aspect, a method of increasing insulin sensitivity in a patient in need thereof increases the patient's HOMA2-S value by between 0.1% and 40% compared to the baseline HOMA2-S value between 0.1 and 6 hours, between 0.1 and 5 hours, between 0.1 and 4 hours, between 0.1 and 3 hours, between 0.1 and 2 hours, or between 0.1 and 1 hour after administration of an effective dose of MANP. In an aspect, a method of increasing insulin sensitivity in a patient in need thereof increases the patient's HOMA2-S value by between 0.1% and 40% compared to the baseline HOMA2-S value between 1 and 6 hours, between 2 and 6 hours, between 3 and 6 hours, between 4 and 6 hours, between 5 and 6 hours, between 2 and 5 hours, between 3 and 4 hours, between 1 and 3 hours, between 2 and 4 hours, or between 3 and 5 hours after administration of an effective dose of MANP.

[0200] In an aspect, a method of increasing insulin sensitivity in a patient in need thereof decreases the patient's HOMA2-insulin resistance (HOMA2-IR) value by between 0.1 and 0.6 compared to the baseline HOMA2-IR value within 4 hours after administration of an effective dose of MANP. In an aspect, a method of increasing insulin sensitivity in a patient in need thereof decreases the patient's HOMA2-IR value by between 0.1 and 1, between 0.1 and 0.8, between 0.1 and 0.6, between 0.1 and 0.4, or between 0.1 and 0.2 compared to the baseline HOMA2-IR value within 4 hours after administration of an effective dose of MANP. In an aspect, a method of increasing insulin sensitivity in a patient in need thereof decreases the patient's HOMA2-IR value by between 0.2 and 1, between 0.4 and 1, between 0.6 and 1, or between 0.8 and 1 compared to the baseline HOMA2-IR value within 4 hours after administration of an effective dose of MANP. In an aspect, a method of increasing insulin sensitivity in a patient in need thereof decreases the patient's HOMA2-IR value by between 0.2 and 0.8, between 0.4 and 0.6, between 0.2 and 0.6, or between 0.4 and 0.8 compared to the baseline HOMA2-IR value within 4 hours after administration of an effective dose of MANP.

[0201] In an aspect, a method of increasing insulin sensitivity in a patient in need thereof reduces the patient's HOMA2-IR value by between 0.1 and 1 as compared to the baseline HOMA2-IR value within 0.1 hour, 0.2 hour, 0.3 hour, 0.4 hour, 0.5 hour, 0.6 hour, 0.7 hour, 0.8 hour, 0.9 hour, 1 hour, 1.5 hour, 2 hour, 2.5 hour, 3 hour, 3.5 hour, 4 hour, 4.5 hour, 5 hour, 5.5 hour, 6 hour, 6.5 hour, 7 hour, 7.5 hour, 8 hour, 8.5 hour, 9 hour, 9.5 hour, 10 hour, 12 hour, or 24 hour after administration of an effective dose of MANP. In an aspect, a method of increasing insulin sensitivity in a patient in need thereof reduces the patient's HOMA2-IR value by between 0.1 and 1 as compared to the baseline HOMA2-IR value between 0.1 and 6 hours, between 0.1 and 5 hours, between 0.1 and 4 hours, between 0.1 and 3 hours, between 0.1 and 2 hours, or between 0.1 and 1 hour after administration of an effective dose of MANP. In an aspect, a method of increasing insulin sensitivity in a patient in need thereof reduces the patient's HOMA2-IR value by between 0.1 and 1 as compared to the baseline HOMA2-IR value between 1 and 6 hours, between 2 and 6 hours, between 3 and 6 hours, between 4 and 6 hours, between 5 and 6 hours, between 2 and 5 hours, between 3 and 4 hours, between 1 and 3 hours, between 2 and 4 hours, or between 3 and 5 hours after administration of an effective dose of MANP.

[0202] In an aspect, the HOMA2-S value and the HOMA2-IR value are calculated by the HOMA2 calculator software published by the Oxford Centre for Diabetes, Endocrinology and Metabolism. See Wallace et al. (2004), Diabetes Care., 1487-1495.

[0203] In one aspect, a method of increasing insulin sensitivity in a patient in need thereof includes parenterally administering an effective amount of MANP at a dose of about 0.01 ng polypeptide / kg to about 100 mg polypeptide / kg body weight (e.g., about 10 ng polypeptide / kg to about 50 mg polypeptide / kg, about 20 ng polypeptide / kg to about 10 mg polypeptide / kg, about 0.1 ng polypeptide / kg to about 20 ng polypeptide / kg, about 3 ng polypeptide / kg to about 10 ng polypeptide / kg, about 10 ng polypeptide / kg to about 50 ng polypeptide / kg, about 50 ng polypeptide / kg to about 100 μg polypeptide / kg, about 1 μg polypeptide / kg to about 10 μg polypeptide / kg, about 1 μg polypeptide / kg to about 5 μg polypeptide / kg, about 3 μg polypeptide / kg to about 7 μg polypeptide / kg, about 5 μg polypeptide / kg to about 10 μg polypeptide / kg, about 10 μg polypeptide / kg to about 20 μg polypeptide / kg, or about 20 μg polypeptide / kg to about 100 μg polypeptide / kg), although other doses can provide beneficial results. In one aspect, a composition comprising MANP can be administered parenterally (e.g., by injection, such as subcutaneous injection, or by oral ingestion) at a dose of about 0.5 ng polypeptide to about 20 g polypeptide (e.g., about 500 ng polypeptide to about 10 g polypeptide, about 1 μg polypeptide to about 2 g polypeptide, about 5 ng polypeptide to about 4 μg polypeptide, about 150 ng polypeptide to about 2 μg polypeptide, about 0.5 μg polypeptide to about 10 μg polypeptide, about 2.5 μg polypeptide to about 20 mg polypeptide, about 50 μg polypeptide to about 2 mg polypeptide, about 50 μg polypeptide to about 1 mg polypeptide, about 150 μg polypeptide to about 1.4 mg polypeptide, about 250 μg polypeptide to about 2 mg polypeptide, about 0.5 mg polypeptide to about 4 mg polypeptide, or about 1 mg polypeptide to about 20 mg polypeptide), although other doses can provide beneficial results. In one aspect, the MANP can be administered by infusion at a dose of, for example, about 1 pmol / kg / minute to about 500 nmol / kg / minute (e.g., about 1 to 10 pmol / kg / minute, about 10 to 100 pmol / kg / minute, about 100 to 300 pmol / kg / minute, about 250 to 500 pmol / kg / minute, about 500 pmol / kg / minute to about 1 nmol / kg / minute, about 1 to 10 nmol / kg / minute, about 10 to 100 nmol / kg / minute, about 100 to 300 nmol / kg / minute, or about 250 to 500 nmol / kg / minute).

[0204] In an aspect, a method of increasing plasma NEFA in a patient in need thereof comprises parenterally administering to the patient an effective dose of MANP. In an aspect, a method of increasing plasma NEFA in a patient in need thereof increases plasma NEFA by 1 mM compared to baseline plasma NEFA within 1 hour after administration of an effective dose of MANP. In an aspect, a method of increasing plasma NEFA in a patient in need thereof increases plasma NEFA by between 0.01 and 2 mM, between 0.01 and 1.5 mM, between 0.01 and 0.5 mM, between 0.01 and 0.2 mM, or between 0.01 and 0.1 mM compared to baseline plasma NEFA within 1 hour after administration of an effective dose of MANP. In an aspect, a method of increasing plasma NEFA in a patient in need thereof increases plasma NEFA by between 0.1 and 2 mM, between 0.2 and 2 mM, between 0.5 and 2 mM, between 1 and 2 mM, or between 1.5 and 2 mM compared to baseline plasma NEFA within 1 hour after administration of an effective dose of MANP. In an aspect, a method of increasing plasma NEFA in a patient in need thereof increases plasma NEFA by between 0.1 and 1.5 mM, between 0.2 and 1 mM, between 0.5 and 1 mM, between 0.1 and 0.5 mM, between 0.2 and 1 mM, or between 0.5 and 1.5 mM compared to baseline plasma NEFA within 1 hour after administration of an effective dose of MANP.

[0205] In an aspect, a method of increasing plasma NEFA in a patient in need thereof increases plasma NEFA between 0.01 and 2 μΜ compared to baseline plasma NEFA within 0.1 hour, 0.2 hour, 0.3 hour, 0.4 hour, 0.5 hour, 0.6 hour, 0.7 hour, 0.8 hour, 0.9 hour, 1 hour, 1.5 hour, 2 hour, 2.5 hour, 3 hour, 3.5 hour, 4 hour, 4.5 hour, 5 hour, 5.5 hour, 6 hour, 6.5 hour, 7 hour, 7.5 hour, 8 hour, 8.5 hour, 9 hour, 9.5 hour, 10 hour, 12 hour, or 24 hour after administration of an effective dose of MANP. In an aspect, a method of increasing plasma NEFA in a patient in need thereof increases plasma NEFA between 0.01 and 2 μΜ compared to baseline plasma NEFA between 0.1 and 2 hours, between 0.1 and 1.5 hours, between 0.1 and 1 hour, or between 0.1 and 0.5 hour after administration of an effective dose of MANP. In an aspect, a method of increasing plasma NEFA in a patient in need thereof increases plasma NEFA between 0.01 and 2 μΜ compared to baseline plasma NEFA between 0.5 and 2 hours, between 1 and 2 hours, or between 1.5 and 2 hours after administration of an effective dose of MANP. In an aspect, a method of increasing plasma NEFA in a patient in need thereof increases plasma NEFA between 0.01 and 2 μΜ compared to baseline plasma NEFA between 0.5 and 1.5 hours, between 0.5 and 1 hour, or between 1 and 1.5 hours after administration of an effective dose of MANP.

[0206] In an aspect, plasma NEFA is determined as an indicator of lipolysis and is measured after an overnight fast using a commercial kit (Zem-Bio, Inc., Durham, NC) according to the manufacturer’s instructions.

[0207] In one aspect, a method of increasing plasma NEFA in a patient in need thereof comprises parenterally administering an effective dose of MANP at a dose of about 0.01 ng polypeptide / kg to about 100 mg polypeptide / kg body weight (e.g., about 10 ng polypeptide / kg to about 50 mg polypeptide / kg, about 20 ng polypeptide / kg to about 10 mg polypeptide / kg, about 0.1 ng polypeptide / kg to about 20 ng polypeptide / kg, about 3 ng polypeptide / kg to about 10 ng polypeptide / kg, about 10 ng polypeptide / kg to about 50 ng polypeptide / kg, about 50 ng polypeptide / kg to about 100 μg polypeptide / kg, about 1 μg polypeptide / kg to about 10 μg polypeptide / kg, about 1 μg polypeptide / kg to about 5 μg polypeptide / kg, about 3 μg polypeptide / kg to about 7 μg polypeptide / kg, about 5 μg polypeptide / kg to about 10 μg polypeptide / kg, about 10 μg polypeptide / kg to about 20 μg polypeptide / kg, or about 20 μg polypeptide / kg to about 100 μg polypeptide / kg), although other doses can provide beneficial results. In one aspect, a composition comprising MANP can be administered parenterally (e.g., by injection, such as subcutaneous injection, or by oral ingestion) at a dose of about 0.5 ng polypeptide to about 20 g polypeptide (e.g., about 500 ng polypeptide to about 10 g polypeptide, about 1 μg polypeptide to about 2 g polypeptide, about 5 ng polypeptide to about 4 μg polypeptide, about 150 ng polypeptide to about 2 μg polypeptide, about 0.5 μg polypeptide to about 10 μg polypeptide, about 2.5 μg polypeptide to about 20 mg polypeptide, about 50 μg polypeptide to about 2 mg polypeptide, about 50 μg polypeptide to about 1 mg polypeptide, about 150 μg polypeptide to about 1.4 mg polypeptide, about 250 μg polypeptide to about 2 mg polypeptide, about 0.5 mg polypeptide to about 4 mg polypeptide, or about 1 mg polypeptide to about 20 mg polypeptide), although other doses can provide beneficial results. In one aspect, MANP can be administered by infusion at a dose of, e.g., about 1 pmol / kg / min to about 500 nmol / kg / min (e.g., about 1 to 10 pmol / kg / min, about 10 to 100 pmol / kg / min, about 100 to 300 pmol / kg / min, about 250 to 500 pmol / kg / min, about 500 pmol / kg / min to about 1 nmol / kg / min, about 1 to 10 nmol / kg / min, about 10 to 100 nmol / kg / min, about 100 to 300 nmol / kg / min, or about 250 to 500 nmol / kg / min).

[0208] In an aspect, the methods of the present disclosure include administering an effective dose of MANP once (e.g., by implanting or injecting a depot composition), or more than once (e.g., by repeated injection or oral administration). When administered more than once, the frequency of administration can range from once or more times per day (e.g., once, twice, three times, four times, or more times per day) to about once every other month (e.g., three to five times per week, about once per week, about twice per month, about once per month, or about once every other month). In an aspect, the methods of the present disclosure include administering MANP via a first route (e.g., intravenously) for a first period of time, and then can be administered via another route (e.g., subcutaneously) for a second period of time. In an aspect, the methods of the present disclosure include intravenously administering MANP to a patient (e.g., a human) at a dose of about 1 pmol / kg / minute to about 500 nmol / kg / minute for 1 hour to 7 days (e.g., 1 to 2 hours, 2 to 4 hours, 4 to 6 hours, 6 to 8 hours, 8 to 12 hours, 12 to 24 hours, 24 to 48 hours, 48 to 36 hours, 3 to 4 days, 4 to 4 days, 4 to 6 days, or 6 to 7 days), and subsequently can be administered subcutaneously to the patient at a dose of about 0.01 ng / kg to about 100 mg / kg, one to three times per day, for 5 to 30 days or more (e.g., 5 to 7 days, 7 to 10 days, 10 to 14 days, 14 to 21 days, 21 to 28 days, 28 to 30 days, or more than 30 days).

[0209] An effective dose of MANP (or nucleic acid encoding MANP) administered to a patient is an amount sufficient to change a selected parameter (e.g., BP) by at least 10%. In an aspect, an effective dose of MANP is an amount that reduces the BP of a patient identified as having hypertension by at least 10% (e.g., at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 90%, or at least 100%) compared to the patient’s BP prior to administration of the MANP. In an aspect, an effective dose of MANP is an amount that reduces the BP of a patient identified as having hypertension by 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 10% to 90%, 10% to 80%, 10% to 70%, 10% to 60%, 10% to 50%, 10% to 40%, 10% to 30%, 10% to 20%, 20% to 90%, 30% to 80%, 40% to 60%, 20% to 40%, 30% to 50%, 40% to 60%, 50% to 70%, 60% to 80%, or 70% to 90% compared to the patient’s BP prior to administration of the MANP.

[0210] In one aspect, an effective dose of a MANP is an amount sufficient to increase the patient's plasma cGMP level by at least 10% (e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100%) compared to the patient's plasma cGMP level prior to treatment. In one aspect, an effective dose of a MANP is an amount sufficient to increase the patient's plasma cGMP level by 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 10% to 90%, 10% to 80%, 10% to 70%, 10% to 60%, 10% to 50%, 10% to 40%, 10% to 30%, 10% to 20%, 20% to 90%, 30% to 80%, 40% to 60%, 20% to 40%, 30% to 50%, 40% to 60%, 50% to 70%, 60% to 80%, or 70% to 90% compared to the patient's plasma cGMP level prior to treatment. In one aspect, an effective dose of a MANP is an amount sufficient to increase the patient's plasma ANP-like peptide level by at least 10% (e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100%) compared to the patient's plasma ANP-like peptide level prior to treatment. In one aspect, an effective dose of a MANP is an amount sufficient to increase the patient's plasma ANP-like peptide level by 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 10% to 90%, 10% to 80%, 10% to 70%, 10% to 60%, 10% to 50%, 10% to 40%, 10% to 30%, 10% to 20%, 20% to 90%, 30% to 80%, 40% to 60%, 20% to 40%, 30% to 50%, 40% to 60%, 50% to 70%, 60% to 80%, or 70% to 90% compared to the patient's plasma ANP-like peptide level prior to treatment.In one aspect, an effective dose of MANP is an amount sufficient to decrease plasma glucose levels by at least 10% (e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100%) compared to the plasma glucose levels of the same patient prior to treatment. In one aspect, an effective dose of MANP is an amount sufficient to decrease plasma glucose levels by 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 10% to 90%, 10% to 80%, 10% to 70%, 10% to 60%, 10% to 50%, 10% to 40%, 10% to 30%, 10% to 20%, 20% to 90%, 30% to 80%, 40% to 60%, 20% to 40%, 30% to 50%, 40% to 60%, 50% to 70%, 60% to 80%, or 70% to 90% compared to the plasma glucose levels of the same patient prior to treatment. In one aspect, an effective dose of MANP is an amount sufficient to increase insulin sensitivity by at least 10% (e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100%) compared to the insulin sensitivity of the same patient prior to treatment. In one aspect, an effective dose of MANP is an amount sufficient to increase insulin sensitivity by 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 10% to 90%, 10% to 80%, 10% to 70%, 10% to 60%, 10% to 50%, 10% to 40%, 10% to 30%, 10% to 20%, 20% to 90%, 30% to 80%, 40% to 60%, 20% to 40%, 30% to 50%, 40% to 60%, 50% to 70%, 60% to 80%, or 70% to 90% compared to the insulin sensitivity of the same patient prior to treatment. In one aspect, an effective dose of MANP is an amount sufficient to increase plasma NEFA by at least 10% (e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100%) compared to the plasma NEFA levels of the same patient prior to treatment.In one aspect, an effective dose of MANP is an amount sufficient to increase plasma NEFA by 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 10% to 90%, 10% to 80%, 10% to 70%, 10% to 60%, 10% to 50%, 10% to 40%, 10% to 30%, 10% to 20%, 20% to 90%, 30% to 80%, 40% to 60%, 20% to 40%, 30% to 50%, 40% to 60%, 50% to 70%, 60% to 80%, or 70% to 90% compared to the patient's plasma NEFA level prior to treatment.

[0211] In one aspect, the amount and frequency of MANP administration to a patient can be titrated, for example, to identify the dose that is most effective for treating hypertension with metabolic syndrome while having the least amount of adverse reactions. If a patient does not respond to a particular amount, that amount can be increased, for example, by a factor of two, three, five, or ten. After receiving this higher concentration, the patient can be monitored for both responsiveness to treatment and toxic symptoms, and the dose and / or frequency of administration can be adjusted accordingly. The effective dose can remain constant, or it can be adjusted as a sliding scale or variable dose depending on the patient's response to treatment.

[0212] In one aspect, the amount and frequency of MANP administration to a patient can be titrated, for example, to identify the dose that is most effective for lowering blood pressure while having the least amount of adverse reactions. If a patient does not respond to a particular amount, that amount can be increased, for example, by a factor of two, three, five, or ten. After receiving this higher concentration, the patient can be monitored for both responsiveness to treatment and toxic symptoms, and the dose and / or frequency of administration can be adjusted accordingly. The effective dose can remain constant, or it can be adjusted as a sliding scale or variable dose depending on the patient's response to treatment.

[0213] In one aspect, the amount and frequency of MANP administration to a patient can be titrated, for example, to identify the dose that is most effective for increasing plasma ANP-like peptides while having the least amount of adverse reactions. If a patient does not respond to a particular amount, that amount can be increased, for example, by a factor of two, three, five, or ten. After receiving this higher concentration, the patient can be monitored for both responsiveness to treatment and toxic symptoms, and the dose and / or frequency of administration can be adjusted accordingly. The effective dose can remain constant, or it can be adjusted as a sliding scale or variable dose depending on the patient's response to treatment.

[0214] In an aspect, the amount and frequency of MANP administration to a patient can be titrated, e.g., to identify the dose that is most effective at reducing plasma glucose levels while having the least amount of adverse reactions. If a patient does not respond to a particular amount, the amount can be increased, e.g., by a factor of two, three, five, or ten. After receiving the higher concentration, the patient can be monitored for both responsiveness to the treatment and toxic symptoms, and the dose and / or frequency of administration can be adjusted accordingly. The effective dose can remain constant, or it can be adjusted as a sliding scale or variable dose depending on the patient's response to the treatment.

[0215] In an aspect, the amount and frequency of MANP administration to a patient can be titrated, e.g., to identify the dose that is most effective at increasing insulin sensitivity while having the least amount of adverse reactions. If a patient does not respond to a particular amount, the amount can be increased, e.g., by a factor of two, three, five, or ten. After receiving the higher concentration, the patient can be monitored for both responsiveness to the treatment and toxic symptoms, and the dose and / or frequency of administration can be adjusted accordingly. The effective dose can remain constant, or it can be adjusted as a sliding scale or variable dose depending on the patient's response to the treatment.

[0216] In an aspect, the amount and frequency of MANP administration to a patient can be titrated, e.g., to identify the dose that is most effective at increasing plasma NEFA while having the least amount of adverse reactions. If a patient does not respond to a particular amount, the amount can be increased, e.g., by a factor of two, three, five, or ten. After receiving the higher concentration, the patient can be monitored for both responsiveness to the treatment and toxic symptoms, and the dose and / or frequency of administration can be adjusted accordingly. The effective dose can remain constant, or it can be adjusted as a sliding scale or variable dose depending on the patient's response to the treatment.

[0217] In an aspect, the frequency of administration can be any frequency that reduces symptoms of, e.g., cardiovascular or cardio-renal disease, in a patient without producing significant toxicity in the patient. In an aspect, the frequency of administration can be about four times per day to about once every 3 months, or about twice per day to about once every other month, or about once per day to about once per month, or about once every other day to about once per week. Further, the frequency of administration can remain constant or can be variable over the duration of treatment. As with the effective dose, various factors can influence the actual frequency of administration for a particular application. In an aspect, the effective dose, duration of treatment, route of administration, and severity of kidney disease can require an increase or decrease in the frequency of administration.

[0218] The effective duration of administration can be any duration that reduces the symptoms of hypertension or metabolic syndrome in the patient without producing significant toxicity in the patient. In one aspect, the effective duration of administration can be any duration that reduces blood pressure in the patient without producing significant toxicity. In one aspect, the effective duration of administration can be any duration that increases plasma ANP-like peptide in the patient without producing significant toxicity. In one aspect, the effective duration of administration can be any duration that reduces plasma glucose levels in the patient without producing significant toxicity. In one aspect, the effective duration of administration can be any duration that increases insulin sensitivity in the patient without producing significant toxicity. In one aspect, the effective duration of administration can be any duration that increases plasma NEFA in the patient without producing significant toxicity. The effective duration can range from one day to several days, to several weeks, months or years. Typically, the effective duration can range from a duration of several days to several months. In one aspect, the effective duration can range from about 1 to 2 weeks to about 36 months or even longer. In one aspect, the treatment can last the lifetime of the individual patient. A variety of factors can influence the actual effective duration for a particular treatment regimen. In one aspect, the effective duration can vary with the frequency of administration, the amount administered, the route of administration, and the severity of the condition.

[0219] Following administration of a MANP as described herein to a patient, the patient can be monitored to determine if the condition has improved. In one aspect, the patient can be monitored for several hours, days, months, or years following administration of a MANP as described herein. In one aspect, patient characteristics that can be monitored include systolic BP, diastolic BP, plasma cGMP, plasma ANP-like peptide, plasma glucose levels, insulin sensitivity, and plasma NEFA.

[0220] In one aspect, the patient can be assessed following treatment to determine if one or more symptoms of the patient's condition have been reduced (e.g., if the patient's BP has been lowered). If the patient does not respond to a particular dose of the MANP, the amount can be increased, for example, by a factor of two, three, five, or ten. Following receipt of this higher concentration, the patient can be monitored for both responsiveness to the treatment and for signs of toxicity, and adjusted accordingly. The effective dose can remain constant, or can be adjusted as a sliding scale or variable dose depending on the patient's response to the treatment.

[0221] The application will be further described in the following examples, which do not limit the scope of the application described in the claims.

[0222] Example

[0223] Example 1: Study design and patients for a study investigating the role of MANP in subjects with hypertension with metabolic syndrome

[0224] Research Design

[0225] like Figure 2 As shown, the study consisted of a screening visit (defined as the first visit on day 0, see details below), a 7-day period of dietary and fluid restriction, and medication. A total of 22 patients with hypertension and metabolic syndrome were enrolled. On day 1, all subjects began a moderate sodium (3.0 g / day) diet and restricted fluid intake to 2.5 L / day for 7 days until discharge from the clinic. On day 5, subjects were admitted to the clinic (second visit), and urinary sodium levels were assessed to confirm adherence to the sodium diet. On day 6, 17 subjects received a single subcutaneous injection of MANP (2.5 μg / kg subcutaneously), and 5 subjects received a placebo (0.9% saline). All patients were observed within 24 hours of MANP or placebo administration. Blood pressure and heart rate were measured at baseline and at several time intervals after MANP or placebo administration, and blood samples were collected. On day 7, after completing the 24-hour observation period, subjects underwent a physical examination and were discharged from the clinic. Subjects discontinued their antihypertensive medication only on days 6 and 7, and continued until discharge from the clinic. Laboratory tests were performed and assessed seven days after the MANP / placebo injection. Subjects were contacted by telephone to review laboratory results and assess the occurrence of any potential side effects following MANP / placebo administration.

[0226] patient

[0227] Hypertension is defined as a sitting systolic blood pressure of 140 mmHg ≤ 180 mmHg or a diastolic blood pressure of 90 mmHg ≤ 100 mmHg, despite the use of at least one antihypertensive medication. Metabolic syndrome is defined according to the National Cholesterol Education Program (NCEP) Adult Treatment Group III (ATPIII) criteria. Specifically, metabolic syndrome is defined as the presence of any two of the following characteristics:

[0228] 1. Abdominal obesity is defined as a waist circumference ≥102cm (40 inches) for men and ≥88cm (35 inches) for women;

[0229] 2. Serum triglycerides ≥150 mg / dL (1.7 mmol / L);

[0230] 3. Men with serum high-density lipoprotein (HDL) cholesterol <40 mg / dL (1 mmol / L) and women with <50 mg / dL (1.3 mmol / L), or receiving medication for low HDL cholesterol; and

[0231] 4. Fasting plasma glucose ≥100 mg / dL (5.6 mmol / L), or receiving medication for elevated blood glucose.

[0232] Detailed inclusion criteria are as follows:

[0233] 1. Sitting high blood pressure 140 mm Hg < systolic BP < 180 mm Hg or 90 mm Hg < diastolic BP < 100

[0234] mm Hg, despite receiving anti-hypertensive medication therapy;

[0235] 2. Presence of metabolic syndrome with any two of the following:

[0236] a) Abdominal obesity, male waist circumference > 102 cm (40 inches) and female waist circumference > 88 cm (35 inches);

[0237] b) Serum triglycerides > 150 mg / dL (1.7 mmol / L);

[0238] c) Male serum HDL cholesterol < 40 mg / dL (1 mm / L) and female < 50 mg / dL (1.3 mmol / L), or receiving medication for low HDL cholesterol; and

[0239] d) Fasting plasma glucose > 100 mg / dL (5.6 mmol / L), or receiving medication for elevated blood glucose;

[0240] 3. Age between 18 and 75 years;

[0241] 4. Use of anti-hypertensive medication at a stable dose for 30 days prior to the screening visit; and

[0242] 5. Use of statins, ezetimibe, or a combination thereof at a stable dose for 60 days prior to the screening visit.

[0243] Detailed exclusion criteria are as follows:

[0244] 1. Known hypersensitivity or anaphylaxis to MANP or its components, carperitide, other natriuretic peptides, or related compounds;

[0245] 2. Women who are pregnant or lactating;

[0246] 3. Received any investigational drug or device within 30 days prior to entering the study;

[0247] 4. (Within the past 2 years) History of alcohol abuse, illicit drug use, severe psychiatric illness, physical dependence on any opioid drug, or any history of drug abuse or addiction;

[0248] 5. History of difficulty donating blood, or donation of blood or blood products within 45 days prior to enrollment;

[0249] 6. In the opinion of the investigator, had a clinically significant new illness within 1 month prior to screening;

[0250] 7. History of severe allergies;

[0251] 8. History of coronary artery disease, cerebrovascular disease, or syncope;

[0252] 9. History of epilepsy or other seizure disorder;

[0253] 10. History of organ transplant;

[0254] 11. Malignancy within 5 years prior to the screening visit;

[0255] 12. History of clinically significant endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery;

[0256] and

[0257] 13. Consumption of phosphodiesterase type 5 inhibitors (sildenafil, vardenafil, or tadalafil) within 72 hours of receiving MANP.

[0258] Subjects who met the inclusion criteria based on their electronic medical record were invited via mail with a letter describing the study and asking for their willingness to participate. At the screening visit, medical history was collected, a physical examination was performed, and laboratory test history was screened. An electrocardiogram, BP, and heart rate were recorded, and screening laboratory tests were performed. Verification of BP inclusion criteria was obtained by the mean of 3 sitting BP measurements.

[0259] Randomization to double-blind treatment

[0260] Randomization code was used to assign treatment using a statistician directly sending the code to the Mayo Research Pharmacy. The study pharmacist was blinded to the data analysis and assisted in the preparation of MANP and placebo. A total of 22 patients were randomized to receive either placebo or MANP as a single subcutaneous injection.

[0261] Statistical methods

[0262] Data from patients at baseline and after injection of MANP or placebo were summarized. Normality of the distribution of data was checked, and no data were excluded from the analysis. All adverse events were summarized as counts (percentages). Continuous variables were presented as mean ± standard deviation or median (interquartile range (IQR)). For pharmacokinetic and pharmacodynamic data, repeated measures analysis of variance (ANOVA) was used to compare the main group effect between placebo and MANP and to assess group differences at specific time points. Comparisons to baseline levels were also performed within individual groups. Multiple comparison corrections were performed using the Scheffe method. Pharmacokinetic and pharmacodynamic analyses were performed using the R package (PKNCA).

[0263] C max Determined from the maximum observed during the 24 hours post-treatment. Mean change in systolic and diastolic BP was determined by the average change in systolic and diastolic BP from baseline values occurring over the 24 hours post-treatment. These MANP response indicators were analyzed between genders using unpaired t-tests assuming unequal variances. Correlations between these MANP response indicators and continuous baseline variables were assessed using Spearman’s rank-order method. All statistical analyses were performed with SAS version 9.4 and Graphpad Prism 9, and a two-sided P < 0.05 was considered statistically significant.

[0264] Example 2: Safety and tolerability of MANP

[0265] Subjects enrolled according to Example 1 were followed up to 7 days after MANP / placebo injection. Safety indicators were checked in each subject throughout the study, including adverse events, vital sign measurements, electrocardiograms, clinical laboratory results, and other safety variables.

[0266] Stopping rules were defined as when two subjects met one or both of the following criteria:

[0267] 1. Development of clinically significant hypotension, defined as a decrease in clinical systolic BP from baseline of > 30 mmHg, or a decrease in sitting systolic BP to < 90 mmHg, or development of dizziness or lightheadedness or visual symptoms lasting

[0268] 5 minutes on repeated BP measurements; or

[0269] 2. Any other safety outcome or adverse event deemed by the investigator to raise safety or tolerability concerns for the dose administered.

[0270] Table 3 summarizes the clinical characteristics of the overall and treatment groups. Of these, 17 were randomized to MANP and 5 were randomized to placebo. Mean systolic BP levels at baseline were 149 ± 8 mmHg and mean diastolic BP levels were 83 ± 11 mmHg in the overall study cohort, despite the fact that all subjects were taking at least one antihypertensive medication. Mean BMI and waist circumference were 36 ± 5 kg / m2and 113 ± 9 cm, respectively, indicating that the cohort was characterized by both generalized and abdominal obesity.

[0271] Table 3: Baseline characteristics of 22 patients with hypertension with metabolic syndrome

[0272]

[0273]

[0274] * eGFR = estimated glomerular filtration rate; BUN = blood urea nitrogen; NT-proBNP = N-terminal prohormone B-type natriuretic peptide; ACEI = angiotensin-converting enzyme inhibitor; ARB = angiotensin receptor blocker; CCB = calcium channel blocker.

[0275] As shown in Table 4, no significant changes in electrocardiogram results were observed during the 24 hours following MANP / placebo subcutaneous injection, nor were there changes in drug-related, clinically relevant safety laboratory parameters. All 22 patients completed treatment and did not meet stopping criteria. Table 5 summarizes all adverse events (AEs) associated with treatment during the 7-day follow-up period following MANP / placebo injection. No serious AEs were observed, and orthostatic hypotension and vasovagal syncope were observed in only two different patients receiving MANP.

[0276] Table 4. Safety laboratory parameters 24 hours after MANP administration

[0277] All (n=22) Placebo (n=5) MANP (n=17) Glucose, mg / dL 130(111,144) 127(110,153) 131(114,147) Triglycerides, mg / dL 132(90,157) 139(112,163) 130(87,155) Blood urea nitrogen, mg / dL 16(13,18) 17(15,18) 15(12,18) Creatinine, mg / dL 0.9(0.7,0.9) 0.9(0.8,1.0) 0.8(0.7,1.0) eGFR, mL / min / 1.73 m 2 ]]> 78±14 74±13 80±14 Aspartate aminotransferase, U / L 24.0(20.5,27.0) 23.0(18.0,26.5) 25.0(22.0,28.0) Alkaline phosphatase, IU / L 72.5(55.0,88.5) 78.0(74.5,90.0) 65.0(53.5,91.5) Alanine aminotransferase, U / L 26.5(20.8,33.3) 28.0(18.0,31.5) 25.0(21.0,33.5) Bilirubin, mg / dL 0.5(0.3,0.6) 0.4(0.3,0.8) 0.5(0.4,0.6) Lactate dehydrogenase, U / L 218(204,222) 230(182,257) 214(204,220) Creatine kinase, IU / L 61(38,97) 79(48,224) 51(38,88) Complete blood count Hemoglobin, g / dL 13.8(13.0,14.9) 13.9(11.5,15.6) 13.7(13.1,14.5) Hematocrit test, % 41.5(38.9,43.7) 42.7(35.9,45.9) 41.4(39.1,43.7) Red blood cells, 10 12 / L]] 4.7(4.3,5.1) 4.9(4.1,5.4) 4.7(4.4,5.1) Mean corpuscular volume, fl 87.6(84.8,89.9) 87.5(84.6,88.7) 87.6(84.5,90.4) Red blood cell distribution width, % 13.1(12.3,14.0) 13.4(13.1,15.8) 12.9(12.3,14.1) Platelet count, 10 3 / mm 3 ]]> 260(216,298) 263(199,321) 257(227,300) Leukocytes, 10 3 / mm 3 ]]> 6.2(5.7,7.3) 6.6(5.9,8.2) 6.1(5.4,7.2) Neutrophils, 10 3 / mm 3 ]]> 3.5(2.8,4.4) 3.7(3.2,5.5) 3.5(2.5,4.3) lymphocytes, 10 3 / mm 3 ]]> 1.7(1.5,2.2) 1.7(1.5,2.6) 1.7(1.4,2.3) Monocytes, 10 3 / mm 3 ]] 0.6(0.4,0.6) 0.6(0.4,0.8) 0.5(0.4,0.6) Eosinophils, 10 3 / mm 3 ]] 0.1(0.1,0.2) 0.2(0.1,0.3) 0.1(0.1,0.2) Alkaliphilic bacteria, 10 3 / mm 3 ]]> 0.05(0.04,0.06) 0.05(0.03,0.07) 0.06(0.04,0.06)

[0278] Table 5. Adverse events

[0279] Placebo (n=5) MANP (n=17) Orthostatic hypotension, n (%) 0(0) 1(6) Vasovagal syncope, n (%) 0(0) 1(6) Mild or slight headache, n (%) 0(0) 0(0) Arrhythmia, n (%) 0(0) 0(0) Second or third degree AV block, n (%) 0(0) 0(0) Ventricular tachycardia >5, n (%) 0(0) 0(0) Ventricular fibrillation or cardiac arrest, n (%) 0(0) 0(0) Tachycardia, n (%) 0(0) 0(0) Paresthesia, n (%) 0(0) 0(0) Dyspnea, n (%) 0(0) 0(0) Gastrointestinal symptoms, n (%) 0(0) 0(0)

[0280] Example 3. Plasma ANP-like peptide and cGMP response to administration of MANP

[0281] To measure plasma ANP-like peptide and cGMP in the subjects of Example 1, blood was drawn into EDTA tubes and centrifuged for 10 minutes at 4°C and 2,500 rpm, then 1 mL of plasma was aliquoted and stored at -80°C until assay. The assay detects both ANP and MANP, which are defined as ANP-like peptides in these examples. The assay has <1% cross-reactivity with NT-proANP, BNP, C-type natriuretic peptide (CNP), endothelin, and adrenomedullin. Plasma cGMP was measured with a commercial ELISA kit (Enzo Life Sciences, Farmingdale, NY) according to the manufacturer's instructions. Both plasma ANP-like peptide and cGMP were measured in samples collected at baseline, 0.5, 1, 2, 4, 6, 12, and 24 hours after administration of MANP or placebo. C max Indicates the highest concentration detected in plasma during the 24-hour follow-up period after MANP administration. max Indicates the time required to reach C max required.

[0282] As shown in Table 6 and Figure 3, in patients receiving MANP, plasma ANP-like peptide increased rapidly and significantly at 30 minutes post-injection (change from baseline: 48.9 ± 19.5 pg / mL, P = 0.03), and elevated levels of plasma ANP-like peptide persisted to 1 hour post-injection (change from baseline: 26.4 ± 12.8 pg / mL, P = 0.06). At the same time, a simultaneous and significant increase in plasma cGMP was also observed in the MANP group at 30 minutes (change from baseline: 4.8 ± 2.0 pmol / mL, P = 0.02) and 1 hour (change from baseline: 2.9 ± 1.3 pmol / mL, P = 0.03) post-administration; the trend of increase persisted to nearly 4 hours post-injection. There were no significant changes in plasma ANP-like peptide or cGMP levels in the placebo group compared to baseline.

[0283] Table 6. Pharmacokinetic and pharmacodynamic indices of MANP in subjects with hypertension and metabolic syndrome

[0284] C max (pg / mL) T max (min)]]> C max (pmol / mL) T max (min)]]> ANP-like peptide 84.0(49.7,165.8) 45(30,60) cGMP 12.7(10.1,20.3) 30(30,240)

[0285] Example 4. Response of sitting BP to administration of MANP

[0286] To measure BP and heart rate in the subjects of Example 1, BP and heart rate were monitored continuously on the day of administration of the study drug (MANP or placebo). Sitting BP and heart rate were recorded prior to injection of MANP or placebo and at 0.5, 1, 2, 4, 6, 12, and 24 hours after administration of MANP or placebo.

[0287] As shown in Figure 4, in the MANP group, sitting systolic BP decreased compared to baseline, with the maximum decrease occurring at 6 hours post-injection (change from baseline: -5.7 ± 2.9 mmHg, P = 0.06). Systolic BP remained below baseline 12 hours after MANP dosing. In contrast, no decrease in systolic BP was observed in the placebo group. Sitting diastolic BP tended to be lower than baseline at 6 hours post- administration of MANP (change from baseline: -2.2 ± 1.2 mmHg, P = 0.09). A slight and statistically insignificant increase in heart rate was observed in both groups at 6 hours post-injection and thereafter. There was no difference in heart rate between the MANP group and the placebo group at any time point.

[0288] Example 5. Individual response to MANP

[0289] The response to MANP in the subjects of Example 1 was evaluated by the following during the 24-hour follow-up period after injection of MANP: (1) C max max of plasma ANP-like peptide, max(3) mean change in systolic BP and (4) mean change in diastolic BP. Results are shown in Table 7.

[0290] Although an increase in plasma ANP-like peptide was observed in all 17 patients, baseline ANP levels were positively correlated with the C max max of plasma ANP-like peptide after MANP administration (p = 0.53, P = 0.03). Furthermore, the C max max of plasma ANP-like peptide was positively correlated with waist circumference (p = 0.51, P = 0.04), but not with BMI (p = 0.28, P = 0.28). As shown in Figure 6A and Figure 7A , age, gender and baseline cGMP were not correlated with the C max max of plasma ANP-like peptide. At the same time, only 2 out of the 17 patients did not find an increase in cGMP cycling levels at any of the measured time points. As shown in Figure 6B and Figure 7B , the C max max of plasma cGMP was not correlated with age, gender, BMI, waist circumference, baseline plasma ANP and baseline plasma cGMP. Each circle represents one subject, solid circles indicate males and open circles indicate females.

[0291] In the 17 patients receiving MANP, a decrease in systolic BP was observed in all patients and a decrease in diastolic BP was observed in 15. The mean changes in systolic and diastolic BP were -3.4 (IQR: -8.0 - -3.0) mmHg and -2.4 (IQR: -3.3 - 0.7) mmHg, respectively. As shown in Figure 6C and Figure 6D , the mean change in systolic BP was more pronounced in males than in females (P = 0.008), while the mean change in diastolic BP was similar between the two genders (P = 0.25). As shown in Figure 5A and Figure 5BAs shown, mean systolic BP change was positively correlated with baseline plasma cGMP, from which those with lower baseline plasma cGMP levels had greater mean systolic BP reduction within 24 hours after MANP injection (p = 0.66, P = 0.005). Each circle represents a subject, with solid circles indicating males and open circles indicating females. This significant correlation was further verified by repeating the same analysis using median BP changes (baseline cGMP vs. median systolic BP change: p = 0.73, P = 0.0008) as shown in Table 8 or maximum BP changes (baseline cGMP vs. maximum systolic BP change: p = 0.59, P = 0.01) as shown in Table 9. For each subject, median BP (systolic or diastolic) change was defined as the median value of BP changes within 0.5, 1, 2, 4, 6, 12, and 24 hours after MANP administration. For each subject, maximum BP (systolic or diastolic) change was defined as the most extreme value of BP changes within 0.5, 1, 2, 4, 6, 12, and 24 hours after MANP administration. Mean diastolic BP change did not appear to be influenced by baseline plasma ANP, baseline plasma cGMP, age, or waist circumference, and had a weak negative trend with BMI (p = -0.44, P = 0.08).

[0292] Table 7. Test and baseline characteristics of the MANP group

[0293]

[0294] Table 8. Correlation between baseline characteristics and median BP changes in the MANP group

[0295]

[0296] Table 9. Correlation between baseline characteristics and maximum BP changes in the MANP group

[0297] Example 6. Metabolic effects of MANP

[0298] To measure plasma metabolic parameters, plasma metabolites including glucose, insulin, NEFA, glycerol, triglyceride, and adiponectin were measured at baseline, 0.5, 1, 2, and 4 hours after MANP or placebo administration after overnight fasting while subjects continued to fast.

[0299] Plasma glucose was measured using the Roche glucose reagent (Indianapolis, IN). Plasma insulin was measured with the Roche insulin assay (Indianapolis, IN), which is a two-site immunoassay using electrochemiluminescence immunoassay “ECLIA” detection. Plasma NEFA and glycerol were determined as markers of lipolysis and measured using commercial kits (Zem-Bio, Inc., Durham, NC) according to the manufacturer’s instructions. Plasma triglycerides were measured by an automated enzymatic method using the Roche triglyceride reagent (Indianapolis, IN) as this method is referenced by the Centers for Disease Control and Prevention (CDC) in the standardization method performed in the Cardiovascular Risk Assessment Laboratory. Plasma adiponectin was measured using a commercial kit (Millipore Sigma, Burlington, MA) according to the manufacturer’s protocol.

[0300] As shown in Table 10, plasma glucose levels did not significantly change in the placebo group, whereas we observed a trend for a decrease in plasma glucose levels 1 hour after MANP administration (change from baseline: -3.1 ± 1.5 mg / mL, P = 0.06), and importantly, a significant decrease in plasma glucose levels 2 hours (change from baseline: -4.7 ± 2.1 mg / mL, P = 0.04) and 4 hours (change from baseline: -13.1 ± 3.9 mg / mL, P = 0.003) after MANP administration. In the placebo group, plasma insulin increased 1 hour and 2 hours after injection, whereas the levels of plasma insulin remained stable in the MANP group. The change in insulin levels reached statistical significance between the MANP and placebo groups 1 hour (P = 0.02) and 2 hours (P = 0.03) after injection. We also observed a trend for an increase in plasma NEFA in the MANP group, and this increase reached statistical significance 1 hour after MANP administration (P = 0.01). There were no significant changes in the levels of plasma glycerol, triglycerides, and adiponectin during the first 4 hours after MANP injection.

[0301] As shown in Table 11, HOMA2 values were calculated for the first 4 hours after injection as indirect and surrogate markers. Before treatment, there were no differences in HOMA2 insulin sensitivity (HOMA2-S) values (P = 0.99) and HOMA2 insulin resistance (HOMA2-IR) values (P = 0.36) between the placebo and MANP groups. From baseline to 4 hours after injection, as shown in Table 11, there was a trend for an increase in HOMA2-S values in the placebo group (P = 0.06) and a significant increase in the MANP group (P = 0.02). There was a significant decrease in HOMA2-IR values in the MANP group (P = 0.02) but not in the placebo group (P = 0.06). Figure 8AAs shown, HOMA2-S values increased overall in the MANP group, but not in the placebo group [median change from baseline in the MANP group: 13.7% (IQR: 4.2%, 32.7%); median change from baseline in the placebo group: -12.4% (IQR: -30.6%, 2.0%)]. Consistent with this, as shown, HOMA2-IR values also decreased in the MANP group, but not in the placebo group [median change from baseline in the MANP group: -0.485 (IQR: -0.682, -0.148); median change from baseline in the placebo group: 0.485 (IQR: -0.148, 1.022)]. Figure 8B As shown, HOMA2-S values increased overall in the MANP group, but not in the placebo group [median change from baseline in the MANP group: 13.7% (IQR: 4.2%, 32.7%); median change from baseline in the placebo group: -12.4% (IQR: -30.6%, 2.0%)]. Consistent with this, as shown, HOMA2-IR values also decreased in the MANP group, but not in the placebo group [median change from baseline in the MANP group: -0.485 (IQR: -0.682, -0.148); median change from baseline in the placebo group: 0.485 (IQR: -0.148, 1.022)].

[0302] Table 10. Plasma levels of metabolites before and after MANP / placebo administration

[0303]

[0304] Table 11. HOMA2 indices before and after MANP / placebo administration

[0305]

[0306] Other Embodiments

[0307] It is to be understood that while the present disclosure has been described in conjunction with the specific embodiments thereof, the foregoing description is intended to illustrate and not limit the scope of the present disclosure. Other aspects, advantages and modifications are within the scope of the following claims.

[0308] Example 1. A method of treating hypertension with metabolic syndrome in a patient in need thereof, the patient having a baseline plasma cyclic guanosine monophosphate (cGMP) of less than 10 pmol / mL, the method comprising, consisting essentially of, or consisting of increasing the plasma cGMP as compared to the baseline by administering to the patient an effective dose of a selective spliced atrial natriuretic peptide (MANP).

[0309] Example 2. The method of Example 1, wherein the baseline plasma cGMP is less than 8 pmol / mL.

[0310] Example 3. The method of Example 1 or 2, wherein the increase is an increase in the plasma cGMP from the baseline of 2 to 8 pmol / mL within 0.5 hours after the administration.

[0311] Example 4. The method of Example 1 or 2, wherein the increase is an increase in the plasma cGMP from the baseline of 1 to 5 pmol / mL within 0.5 to 4 hours after the administration.

[0312] Example 5. The method of any one of examples 1-4, wherein the MANP is administered subcutaneously.

[0313] Example 6. The method of example 5, wherein the effective dose is 0.1 pg / kg to 5 pg / kg.

[0314] Example 7. The method of example 6, wherein the effective dose is 2.5 pg / kg.

[0315] Example 8. The method of example 5, wherein the effective dose is 6.5 pg to 750 pg.

[0316] Example 9. The method of example 8, wherein the effective dose is 162.5 pg to 375 pg.

[0317] Example 10. The method of any one of examples 1-4, wherein the MANP is administered intravenously.

[0318] Example 11. The method of example 10, wherein the effective dose is 10 pmol / kg / minute to 100 nmol / kg / minute.

[0319] Example 12. The method of any one of examples 1-4, wherein the MANP is administered intravenously and subsequently administered subcutaneously.

[0320] Example 13. The method of example 12, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 0.1 pg / kg to 5 pg / kg.

[0321] Example 14. The method of example 13, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 2.5 pg / kg.

[0322] Example 15. The method of example 12, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 6.5 pg to 750 pg.

[0323] Example 16. The method of Example 15, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 162.5 pg to 375 pg.

[0324] Example 17. The method of any one of Examples 1 to 16, wherein the patient has a body mass index (BMI) higher than 25 kg / m 2 .

[0325] Example 18. The method of any one of Examples 1 to 17, wherein the patient has a BMI higher than 30 kg / m 2 .

[0326] Example 19. The method of any one of Examples 1 to 18, wherein the patient has a BMI higher than 35 kg / m 2 .

[0327] Example 20. The method of any one of Examples 1 to 19, wherein the patient presents at least two traits selected from the group consisting of abdominal obesity, high serum triglycerides, low serum high-density lipoprotein (HDL) cholesterol, and high fasting plasma glucose.

[0328] Example 21. The method of Example 20, wherein the abdominal obesity is identified by a waist circumference of the male patient greater than or equal to 102 cm or a waist circumference of the female patient greater than or equal to 88 cm.

[0329] Example 22. The method of Example 20 or 21, wherein the high serum triglycerides are identified by serum triglycerides of 150 mg / dL or above in the patient prior to the administration.

[0330] Example 23. The method of any one of Examples 20 to 22, wherein the low HDL cholesterol is identified by an HDL cholesterol lower than 40 mg / dL in the male patient or lower than 50 mg / dL in the female patient.

[0331] Example 24. The method of any one of Examples 20 to 22, wherein the low HDL cholesterol is identified in the patient who is receiving a drug treatment for low HDL cholesterol to increase HDL cholesterol.

[0332] Example 25. The method of any one of Examples 20 to 24, wherein the high fasting plasma glucose is identified by a fasting plasma glucose of 100 mg / dL or above in the patient.

[0333] Example 26. The method of any one of examples 20-24, wherein the high fasting plasma glucose is identified in the patient who is receiving a medication for elevated blood glucose to lower blood glucose.

[0334] Example 27. The method of any one of examples 1-26, wherein the patient presents with a sitting systolic blood pressure between 140 and 180 mmHg or a sitting diastolic blood pressure between 90 and 100 mmHg prior to the administration.

[0335] Example 28. The method of any one of examples 1-27, wherein the patient is taking an anti-hypertensive medication for 30 days prior to the administration, or is taking a statin, ezetimibe, or a combination thereof for 60 days prior to the administration.

[0336] Example 29. The method of any one of examples 1-28, wherein the patient does not present with any trait selected from the group consisting of:

[0337] a known hypersensitivity or allergic reaction to the MANP or components thereof, capropil, other natriuretic peptides, or related compounds;

[0338] being pregnant or lactating;

[0339] having a history of clinically significant endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery;

[0340] taking a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration;

[0341] having received any investigational drug or device within 30 days prior to the administration;

[0342] having a history of alcohol abuse, illicit drug use, severe psychiatric illness, physical dependence on any opioid drug, or any history of drug abuse or addiction (within 2 years prior to the administration);

[0343] having a history of blood donation difficulties, or donating blood or blood products within 45 days prior to the administration;

[0344] having a clinically significant new illness within 1 month prior to the administration;

[0345] having a history of severe allergies;

[0346] having a history of coronary artery disease, cerebrovascular disease, or syncope;

[0347] having a history of epilepsy or other seizure disorders;

[0348] having a history of organ transplant; and

[0349] has had a malignancy within 5 years prior to the administration.

[0350] Example 30. The method of any one of Examples 1-29, wherein the patient does not present any trait selected from the group consisting of:

[0351] a known hypersensitivity or allergic reaction to the MANP or components thereof, capesantide, other natriuretic peptides or related compounds;

[0352] is pregnant or lactating;

[0353] has a history of clinically significant endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery;

[0354] and

[0355] took a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration.

[0356] Example 31. The method of any one of Examples 1-30, wherein within 12 hours after the administration, the patient’s sitting systolic blood pressure is reduced by 0.1% to 15% compared to the baseline sitting systolic blood pressure.

[0357] Example 32. The method of any one of Examples 1-31, wherein within 6 hours after the administration, the patient’s sitting diastolic blood pressure is reduced by 0.1% to 5% compared to the baseline sitting diastolic blood pressure.

[0358] Example 33. The method of any one of Examples 1-32, wherein within 4 hours after the administration, the patient’s plasma glucose is reduced by 1 to 17 mg / mL compared to the baseline plasma glucose.

[0359] Example 34. The method of any one of Examples 1-33, further comprising, consisting essentially of, or consisting of administering a second drug selected from the group consisting of a diuretic, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

[0360] Example 35. A method of reducing blood pressure in a patient in need thereof, the patient having a baseline plasma cyclic guanosine monophosphate (cGMP) of less than 10 pmol / mL, the method comprising, consisting essentially of, or consisting of increasing plasma cGMP compared to the baseline by administering to the patient an effective dose of a selectively spliced atrial natriuretic peptide (MANP).

[0361] Example 36. The method of Example 35, wherein the baseline plasma cGMP is less than 8 pmol / mL.

[0362] Example 37. The method of Example 35 or 36, wherein the increase is an increase in the plasma cGMP from the baseline of 2 to 8 pmol / mL within 0.5 hours after the administration.

[0363] Example 38. The method of Example 35 or 36, wherein the increase is an increase in the plasma cGMP from the baseline of 1 to 5 pmol / mL within 0.5 to 4 hours after the administration.

[0364] Example 39. The method of any one of Examples 35-38, wherein the decrease is a decrease in the patient’s sitting systolic blood pressure compared to a baseline sitting systolic blood pressure of 0.1% to 15% within 12 hours after the administration.

[0365] Example 40. The method of any one of Examples 35-38, wherein the decrease is a decrease in the patient’s sitting diastolic blood pressure compared to a baseline sitting diastolic blood pressure of 0.1% to 5% within 6 hours after the administration.

[0366] Example 41. The method of any one of Examples 35-40, wherein the MANP is administered subcutaneously.

[0367] Example 42. The method of Example 41, wherein the effective dose is 0.1 pg / kg to 5 pg / kg.

[0368] Example 43. The method of Example 42, wherein the effective dose is 2.5 pg / kg.

[0369] Example 44. The method of Example 41, wherein the effective dose is 6.5 pg to 750 pg.

[0370] Example 45. The method of Example 44, wherein the effective dose is 162.5 pg to 375 pg.

[0371] Example 46. The method of any one of Examples 35-40, wherein the MANP is administered intravenously.

[0372] Example 47. The method of Example 46, wherein the effective dose is 10 pmol / kg / minute to 100 nmol / kg / minute.

[0373] Example 48. The method of any one of Examples 35-40, wherein the MANP is administered intravenously and is subsequently administered subcutaneously.

[0374] Example 49. The method of Example 48, wherein the MANP is intravenously administered at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently subcutaneously administered at a dose of 0.1 pg / kg to 5 pg / kg.

[0375] Example 50. The method of Example 49, wherein the MANP is intravenously administered at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently subcutaneously administered at a dose of 2.5 pg / kg.

[0376] Example 51. The method of Example 48, wherein the MANP is intravenously administered at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently subcutaneously administered at a dose of 6.5 pg to 750 pg.

[0377] Example 52. The method of Example 51, wherein the MANP is intravenously administered at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently subcutaneously administered at a dose of 162.5 pg to 375 pg.

[0378] Example 53. The method of any one of Examples 35-52, wherein the patient has a body mass index (BMI) higher than 25 kg / m2. 2

[0379] Example 54. The method of any one of Examples 35-53, wherein the patient has a BMI higher than 30 kg / m2. 2

[0380] Example 55. The method of any one of Examples 35-54, wherein the patient has a BMI higher than 35 kg / m2. 2

[0381] Example 56. The method of any one of Examples 35-55, wherein the patient presents a sitting systolic blood pressure between 140 mmHg and 180 mmHg or a sitting diastolic blood pressure between 90 mmHg and 100 mmHg prior to the administration.

[0382] Example 57. The method of any one of Examples 35-56, wherein the patient takes an anti-hypertensive medication for 30 days prior to the administration, or takes a statin, ezetimibe, or a combination thereof for 60 days prior to the administration.

[0383] ​​​Embodiment 58. The method of any one of embodiments 35-57, wherein the patient presents at least two traits selected from the group consisting of abdominal obesity, high serum triglycerides, low serum high-density lipoprotein (HDL) cholesterol, and high fasting plasma glucose.

[0384] Embodiment 59. The method of embodiment 58, wherein the abdominal obesity is identified by a waist circumference of the male patient greater than or equal to 102 cm or a waist circumference of the female patient greater than or equal to 88 cm.

[0385] Embodiment 60. The method of embodiment 58 or 59, wherein the high serum triglycerides are identified by serum triglycerides of 150 mg / dL or above in the patient prior to the administration.

[0386] Embodiment 61. The method of any one of embodiments 58-60, wherein the low HDL cholesterol is identified by an HDL cholesterol of less than 40 mg / dL in the male patient or an HDL cholesterol of less than 50 mg / dL in the female patient.

[0387] Embodiment 62. The method of any one of embodiments 58-60, wherein the low HDL cholesterol is identified in the patient who is receiving a drug treatment for low HDL cholesterol to increase HDL cholesterol.

[0388] Embodiment 63. The method of any one of embodiments 58-62, wherein the high fasting plasma glucose is identified by a fasting plasma glucose of 100 mg / dL or above in the patient.

[0389] Embodiment 64. The method of any one of embodiments 58-62, wherein the high fasting plasma glucose is identified in the patient who is receiving a drug treatment for elevated blood glucose to lower blood glucose.

[0390] Embodiment 65. The method of any one of embodiments 35-64, wherein the patient does not present any trait selected from the group consisting of:

[0391] a known hypersensitivity or allergic reaction to the MANP or components thereof, capropamil, other natriuretic peptides, or related compounds;

[0392] pregnancy or lactation;

[0393] a history of clinically significant endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal arteries; ingestion of a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration;

[0394] Received any investigational drug or device within 30 days prior to the administration;

[0395] Has a history of alcohol abuse, illicit drug use, severe psychiatric illness, physiological dependence on any opioid drug, or any history of drug abuse or addiction within 2 years prior to the administration;

[0396] Has a history of difficulty donating blood, or donated blood or blood products within 45 days prior to the administration;

[0397] Has a clinically significant new illness within 1 month prior to the administration;

[0398] Has a history of severe allergies;

[0399] Has a history of coronary artery disease, cerebrovascular disease, or syncope;

[0400] Has a history of epilepsy or other seizure disorder;

[0401] Has a history of organ transplantation; and

[0402] Has a malignancy within 5 years prior to the administration.

[0403] Embodiment 66. The method of any one of embodiments 35-65, wherein the patient does not present any trait selected from the group consisting of:

[0404] Known hypersensitivity or allergic reactions to the MANP or components thereof, capesareide, other natriuretic peptides, or related compounds;

[0405] Is pregnant or lactating;

[0406] Has a history of clinically significant endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery;

[0407] and

[0408] Took a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration.

[0409] Embodiment 67. The method of any one of embodiments 35-66, wherein the patient’s plasma glucose is reduced by 1 to 17 mg / mL compared to baseline plasma glucose within 4 hours after the administration.

[0410] Embodiment 68. The method of any one of embodiments 35-67, further comprising, consisting essentially of, or consisting of administering a second drug selected from the group consisting of a diuretic, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

[0411] Example 69. A method of increasing plasma atrial natriuretic peptide (ANP)-like peptide in a patient in need thereof having a baseline plasma cyclic guanosine monophosphate (cGMP) of less than 10 pmol / mL, the method comprising, consisting essentially of, or consisting of increasing plasma cGMP as compared to the baseline by administering to the patient an effective dose of a selective spliced atrial natriuretic peptide (MANP).

[0412] Example 70. The method of Example 69, wherein the baseline plasma cGMP is less than 8 pmol / mL.

[0413] Example 71. The method of Example 69 or 70, wherein the increasing plasma cGMP increases the plasma cGMP from the baseline by 2 to 8 pmol / mL within 0.5 hours after the administration.

[0414] Example 72. The method of Example 69 or 70, wherein the increasing plasma cGMP increases the plasma cGMP from the baseline by 1 to 5 pmol / mL within 0.5 to 4 hours after the administration.

[0415] Example 73. The method of any one of Examples 69-72, wherein the increasing ANP-like peptide increases the ANP-like peptide from a baseline ANP-like peptide by 1 to 70 pg / mL within 0.5 hours after the administration.

[0416] Example 74. The method of any one of Examples 69-72, wherein the increasing ANP-like peptide increases the ANP-like peptide from a baseline ANP-like peptide by 1 to 40 pg / mL within 1 hour after the administration.

[0417] Example 75. The method of any one of Examples 69-74, wherein the MANP is administered subcutaneously.

[0418] Example 76. The method of Example 75, wherein the effective dose is 0.1 pg / kg to 5 pg / kg.

[0419] Example 77. The method of Example 76, wherein the effective dose is 2.5 pg / kg.

[0420] Example 78. The method of Example 75, wherein the effective dose is 6.5 pg to 750 pg.

[0421] Example 79. The method of Example 78, wherein the effective dose is 162.5 pg to 375 pg.

[0422] Example 80. The method of any one of examples 69-74, wherein the MANP is administered intravenously.

[0423] Example 81. The method of example 80, wherein the effective dose is 10 pmol / kg / minute to 100 nmol / kg / minute.

[0424] Example 82. The method of any one of examples 69-74, wherein the MANP is administered intravenously and subsequently administered subcutaneously.

[0425] Example 83. The method of example 82, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 0.1 pg / kg to 5 pg / kg.

[0426] Example 84. The method of example 83, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 2.5 pg / kg.

[0427] Example 85. The method of example 82, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 6.5 pg to 750 pg.

[0428] Example 86. The method of example 85, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 162.5 pg to 375 pg.

[0429] Example 87. The method of any one of examples 69-86, wherein the patient has a body mass index (BMI) higher than 25 kg / m 2 .

[0430] Example 88. The method of any one of examples 69-87, wherein the patient has a BMI higher than 30 kg / m 2 .

[0431] Example 89. The method of any one of examples 69-88, wherein the patient has a BMI higher than 35 kg / m 2 .

[0432] Example 90. The method according to any one of Examples 69 to 89, wherein the patient presents a sitting systolic blood pressure between 140 mmHg and 180 mmHg or a sitting diastolic blood pressure between 90 mmHg and 100 mmHg prior to the administration.

[0433] Example 91. The method according to any one of Examples 69 to 90, wherein the patient has taken an antihypertensive drug for 30 days prior to the administration, or has taken a statin, ezetimibe, or a combination thereof for 60 days prior to the administration.

[0434] Example 92. The method according to any one of Examples 69 to 91, wherein the patient presents at least two traits selected from the group consisting of: abdominal obesity, high serum triglycerides, low serum high-density lipoprotein (HDL) cholesterol, and high fasting plasma glucose.

[0435] Example 93. According to the method of Example 92, wherein abdominal obesity is identified by a waist circumference greater than or equal to 102 cm for male patients or greater than or equal to 88 cm for female patients.

[0436] Example 94. The method according to Example 92 or 93, wherein the high serum triglycerides are identified by serum triglycerides of 150 mg / dL or higher in the patient prior to the administration.

[0437] Example 95. The method according to any one of Examples 92 to 94, wherein the low HDL cholesterol is identified by HDL cholesterol levels below 40 mg / dL in the male patient or below 50 mg / dL in the female patient.

[0438] Example 96. The method according to any one of Examples 92 to 94, wherein the low HDL cholesterol was identified in the patient who was receiving drug treatment for low HDL cholesterol to increase HDL cholesterol.

[0439] Example 97. The method according to any one of Examples 92 to 96, wherein the high fasting plasma glucose is identified by fasting plasma glucose of 100 mg / dL or more in the patient.

[0440] Example 98. The method according to any one of Examples 92 to 96, wherein the high fasting plasma glucose was identified in the patient who was receiving drug treatment for elevated blood glucose to lower blood glucose.

[0441] Example 99. The method according to any one of Examples 69 to 98, wherein the patient does not exhibit any trait selected from the group consisting of:

[0442] Known hypersensitivity or allergic reactions to the MANP or components thereof, to capesantide, to other natriuretic peptides or related compounds;

[0443] Pregnant or lactating;

[0444] History of clinically significant endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal arteries; ingestion of phosphodiesterase type 5 inhibitors (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration;

[0445] Received any investigational drug or device within 30 days prior to the administration;

[0446] History of alcohol abuse, illicit drug use, severe psychiatric disorder, physiological dependence on any opioid drug, or any history of drug abuse or addiction (within 2 years prior to the administration);

[0447] History of difficulty donating blood, or donation of blood or blood products within 45 days prior to the administration;

[0448] Clinically significant new illness within 1 month prior to the administration;

[0449] History of severe allergies;

[0450] History of coronary artery disease, cerebrovascular disease, or syncope;

[0451] History of epilepsy or other seizure disorders;

[0452] History of organ transplantation; and

[0453] Malignancy within 5 years prior to the administration.

[0454] Embodiment 100. The method of any one of embodiments 69-99, wherein the patient does not present any trait selected from the group consisting of:

[0455] Known hypersensitivity or allergic reactions to the MANP or components thereof, to capesantide, to other natriuretic peptides or related compounds;

[0456] Pregnant or lactating;

[0457] History of clinically significant endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal arteries;

[0458] and

[0459] Ingestion of phosphodiesterase type 5 inhibitors (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration.

[0460] Embodiment 101. The method of any one of embodiments 69-100, wherein within 12 hours after the administration, the patient’s sitting systolic blood pressure is reduced by 0.1% to 15% compared to the baseline sitting systolic blood pressure.

[0461] Embodiment 102. The method of any one of embodiments 69-101, wherein within 6 hours after the administration, the patient’s sitting diastolic blood pressure is reduced by 0.1% to 5% compared to the baseline sitting diastolic blood pressure.

[0462] Embodiment 103. The method of any one of embodiments 69-102, wherein within 4 hours after the administration, the patient’s plasma glucose is reduced by 1 to 17 mg / mL compared to the baseline plasma glucose.

[0463] Embodiment 104. The method of any one of embodiments 69-103, further comprising, consisting essentially of, or consisting of administering a second medicament selected from the group consisting of a diuretic, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

[0464] Embodiment 105. A method of reducing plasma glucose levels in a patient in need thereof, comprising, consisting essentially of, or consisting of administering to the patient an effective dose of a selectively spliced atrial natriuretic peptide (MANP).

[0465] Embodiment 106. The method of embodiment 105, wherein the reduction is a reduction in the plasma glucose levels by 1 to 17 mg / mL compared to the baseline plasma glucose within 4 hours after the administration.

[0466] Embodiment 107. The method of embodiment 105 or 106, wherein the MANP is administered subcutaneously.

[0467] Embodiment 108. The method of embodiment 107, wherein the effective dose is 0.1 pg / kg to 5 pg / kg.

[0468] Embodiment 109. The method of embodiment 108, wherein the effective dose is 2.5 pg / kg.

[0469] Embodiment 110. The method of embodiment 107, wherein the effective dose is 6.5 pg to 750 pg.

[0470] Embodiment 111. The method of embodiment 110, wherein the effective dose is 162.5 pg to 375 pg.

[0471] Example 112. The method of either of Examples 105 or 106, wherein the MANP is administered intravenously.

[0472] Example 113. The method of Example 112, wherein the effective dose is 10 pmol / kg / minute to 100 nmol / kg / minute.

[0473] Example 114. The method of either of Examples 105 or 106, wherein the MANP is administered intravenously and subsequently administered subcutaneously.

[0474] Example 115. The method of Example 114, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 0.1 pg / kg to 5 pg / kg.

[0475] Example 116. The method of Example 115, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 2.5 pg / kg.

[0476] Example 117. The method of Example 114, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 6.5 pg to 750 pg.

[0477] Example 118. The method of Example 117, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 162.5 pg to 375 pg.

[0478] Example 119. The method of any one of Examples 105-118, wherein the patient has a body mass index (BMI) higher than 25 kg / m 2 .

[0479] Example 120. The method of any one of Examples 105-119, wherein the patient has a BMI higher than 30 kg / m 2 .

[0480] Example 121. The method of any one of Examples 105-120, wherein the patient has a BMI higher than 35 kg / m 2 .

[0481] Embodiment 122. The method of any one of embodiments 105-121, wherein the patient presents with a sitting systolic blood pressure of between 140 mmHg and 180 mmHg or a sitting diastolic blood pressure of between 90 mmHg and 100 mmHg prior to the administration.

[0482] Embodiment 123. The method of any one of embodiments 105-122, wherein the patient is taking an anti-hypertensive medication for 30 days prior to the administration, or is taking a statin, ezetimibe, or a combination thereof for 60 days prior to the administration.

[0483] Embodiment 124. The method of any one of embodiments 105-123, wherein the patient presents with at least two traits selected from the group consisting of abdominal obesity, high serum triglycerides, low serum high density lipoprotein (HDL) cholesterol, and high fasting plasma glucose.

[0484] Embodiment 125. The method of embodiment 124, wherein the abdominal obesity is identified by a waist circumference of the male patient of greater than or equal to 102 cm or the female patient of greater than or equal to 88 cm.

[0485] Embodiment 126. The method of embodiment 124 or 125, wherein the high serum triglycerides are identified by serum triglycerides of 150 mg / dL or above in the patient prior to the administration.

[0486] Embodiment 127. The method of any one of embodiments 124-126, wherein the low HDL cholesterol is identified by an HDL cholesterol of less than 40 mg / dL in the male patient or less than 50 mg / dL in the female patient.

[0487] Embodiment 128. The method of any one of embodiments 124-126, wherein the low HDL cholesterol is identified in the patient who is receiving a medication for low HDL cholesterol to increase HDL cholesterol.

[0488] Embodiment 129. The method of any one of embodiments 124-128, wherein the high fasting plasma glucose is identified by a fasting plasma glucose of 100 mg / dL or above in the patient.

[0489] Embodiment 130. The method of any one of embodiments 124-128, wherein the high fasting plasma glucose is identified in the patient who is receiving a medication for elevated blood glucose to decrease blood glucose.

[0490] Embodiment 131. The method of any one of embodiments 105-130, wherein the patient does not present any trait selected from the group consisting of:

[0491] a known hypersensitivity or allergic reaction to the MANP or components thereof, capesareide, other natriuretic peptides or related compounds;

[0492] being pregnant or lactating;

[0493] having a clinically significant endogenous renal disease, history of renal artery stenosis or fibromuscular dysplasia of the renal artery; taking a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration;

[0494] having received any investigational drug or device within 30 days prior to the administration;

[0495] having a history of alcohol abuse, illicit drug use, severe psychiatric illness, physiological dependence on any opioid drug, or any history of drug abuse or addiction (within 2 years prior to the administration);

[0496] having a history of difficulty donating blood, or donating blood or blood products within 45 days prior to the administration;

[0497] having a clinically significant new illness within 1 month prior to the administration;

[0498] having a history of severe allergies;

[0499] having a history of coronary artery disease, cerebrovascular disease, or syncope;

[0500] having a history of epilepsy or other seizure disorders;

[0501] having a history of organ transplant; and

[0502] having a malignancy within 5 years prior to the administration.

[0503] Embodiment 132. The method of any one of embodiments 105-131, wherein the patient does not present any trait selected from the group consisting of:

[0504] a known hypersensitivity or allergic reaction to the MANP or components thereof, capesareide, other natriuretic peptides or related compounds;

[0505] being pregnant or lactating;

[0506] having a clinically significant endogenous renal disease, history of renal artery stenosis or fibromuscular dysplasia of the renal artery;

[0507] and

[0508] consumed within 72 hours prior to the administration of a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil).

[0509] Embodiment 133. The method of any one of embodiments 105-131, wherein within 12 hours after the administration, the patient’s sitting systolic blood pressure is reduced by 0.1% to 15% compared to the baseline sitting systolic blood pressure.

[0510] Embodiment 134. The method of any one of embodiments 105-132, wherein within 6 hours after the administration, the patient’s sitting diastolic blood pressure is reduced by 0.1% to 5% compared to the baseline sitting diastolic blood pressure.

[0511] Embodiment 135. The method of any one of embodiments 105-133, further comprising, consisting essentially of, or consisting of administering a second medicament selected from the group consisting of a diuretic, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

[0512] Embodiment 136. A method of increasing insulin sensitivity in a patient in need thereof, comprising, consisting essentially of, or consisting of administering to the patient an effective dose of a selectively spliced atrial natriuretic peptide (MANP).

[0513] Embodiment 137. The method of embodiment 136, wherein the increasing the insulin sensitivity is increasing the patient’s updated Homeostatic Model Assessment Model Insulin Sensitivity (HOMA2-S) value by 0.1% to 35% compared to a baseline HOMA2-S value within 4 hours after the administration.

[0514] Embodiment 138. The method of embodiment 136, wherein the increasing the insulin sensitivity is decreasing the patient’s updated Homeostatic Model Assessment Model Insulin Resistance (HOMA2-IR) value from a baseline HOMA2-IR value by 0.1 to 0.6 within 4 hours after the administration.

[0515] Embodiment 139. The method of any one of embodiments 136-138, wherein the MANP is administered subcutaneously.

[0516] Embodiment 140. The method of embodiment 139, wherein the effective dose is 0.1 pg / kg to 5 pg / kg.

[0517] Embodiment 141. The method of embodiment 140, wherein the effective dose is 2.5 pg / kg.

[0518] Example 142. The method of Example 139, wherein the effective dose is 6.5 pg to 750 pg.

[0519] Example 143. The method of Example 142, wherein the effective dose is 162.5 pg to 375 pg.

[0520] Example 144. The method of any one of Examples 136-138, wherein the MANP is administered intravenously.

[0521] Example 145. The method of Example 144, wherein the effective dose is 10 pmol / kg / minute to 100 nmol / kg / minute.

[0522] Example 146. The method of any one of Examples 136-138, wherein the MANP is administered intravenously and subsequently administered subcutaneously.

[0523] Example 147. The method of Example 146, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 0.1 pg / kg to 5 pg / kg.

[0524] Example 148. The method of Example 147, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 2.5 pg / kg.

[0525] Example 149. The method of Example 146, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 6.5 pg to 750 pg.

[0526] Example 150. The method of Example 149, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 162.5 pg to 375 pg.

[0527] Example 151. The method of any one of Examples 136-150, wherein the patient has a body mass index (BMI) higher than 25 kg / m 2 .

[0528] Example 152. The method of any one of Examples 136-151, wherein the patient has a BMI higher than 30 kg / m 2 .

[0529] Embodiment 153. The method of any one of embodiments 136-152, wherein the patient has a BMI higher than 35 kg / m 2 .

[0530] Embodiment 154. The method of any one of embodiments 136-153, wherein the patient presents a sitting systolic blood pressure between 140 mmHg and 180 mmHg or a sitting diastolic blood pressure between 90 mmHg and 100 mmHg prior to the administration.

[0531] Embodiment 155. The method of any one of embodiments 136-154, wherein the patient is taking an anti-hypertensive medication for 30 days prior to the administration, or is taking a statin, ezetimibe, or a combination thereof for 60 days prior to the administration.

[0532] Embodiment 156. The method of any one of embodiments 136-155, wherein the patient presents at least two traits selected from the group consisting of abdominal obesity, high serum triglycerides, low serum high-density lipoprotein (HDL) cholesterol, and high fasting plasma glucose.

[0533] Embodiment 157. The method of embodiment 156, wherein the abdominal obesity is identified by a waist circumference greater than or equal to 102 cm for the male patient or greater than or equal to 88 cm for the female patient.

[0534] Embodiment 158. The method of embodiment 156 or 157, wherein the high serum triglycerides are identified by serum triglycerides of 150 mg / dL or above in the patient prior to the administration.

[0535] Embodiment 159. The method of any one of embodiments 156-158, wherein the low HDL cholesterol is identified by an HDL cholesterol of less than 40 mg / dL in the male patient or less than 50 mg / dL in the female patient.

[0536] Embodiment 160. The method of any one of embodiments 156-158, wherein the low HDL cholesterol is identified in the patient who is receiving a medication for low HDL cholesterol to increase HDL cholesterol.

[0537] Embodiment 161. The method of any one of embodiments 156-160, wherein the high fasting plasma glucose is identified by a fasting plasma glucose of 100 mg / dL or above in the patient.

[0538] Embodiment 162. The method of any one of embodiments 156-160, wherein the high fasting plasma glucose is identified in the patient who is receiving a medication for elevated blood glucose to lower blood glucose.

[0539] Embodiment 163. The method of any one of embodiments 136-162, wherein the patient does not present any trait selected from the group consisting of:

[0540] a known hypersensitivity or allergic reaction to the MANP or components thereof, capropil, other natriuretic peptides or related compounds;

[0541] is pregnant or lactating;

[0542] has a history of clinically significant endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal arteries; took a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration;

[0543] received any investigational drug or device within 30 days prior to the administration;

[0544] has a history of alcohol abuse, illicit drug use, severe psychiatric disorder, physiological dependence on any opioid drug, or any history of drug abuse or addiction (within 2 years prior to the administration);

[0545] has a history of blood donation difficulties, or donated blood or blood products within 45 days prior to the administration;

[0546] has a clinically significant new illness within 1 month prior to the administration;

[0547] has a history of severe allergies;

[0548] has a history of coronary artery disease, cerebrovascular disease, or syncope;

[0549] has a history of epilepsy or other seizure disorders;

[0550] has a history of organ transplantation; and

[0551] has a malignancy within 5 years prior to the administration.

[0552] Embodiment 164. The method of any one of embodiments 136-163, wherein the patient does not present any trait selected from the group consisting of:

[0553] a known hypersensitivity or allergic reaction to the MANP or components thereof, capropil, other natriuretic peptides or related compounds;

[0554] a known hypersensitivity or allergic reaction to the MANP or components thereof, capropil, other natriuretic peptides or related compounds;

[0555] is pregnant or lactating;

[0556] has a history of clinically significant endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery;

[0557] and

[0558] took a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration.

[0559] Embodiment 165. The method of any one of embodiments 136-164, wherein within 12 hours after the administration, the patient’s sitting systolic blood pressure is reduced by 0.1% to 15% compared to the baseline sitting systolic blood pressure.

[0560] Embodiment 166. The method of any one of embodiments 136-165, wherein within 6 hours after the administration, the patient’s sitting diastolic blood pressure is reduced by 0.1% to 5% compared to the baseline sitting diastolic blood pressure.

[0561] Embodiment 167. The method of any one of embodiments 136-166, wherein within 4 hours after the administration, the patient’s plasma glucose is reduced by 1 to 17 mg / mL compared to the baseline plasma glucose.

[0562] Embodiment 168. The method of any one of embodiments 136-167, further comprising, consisting essentially of, or consisting of administering a second drug selected from the group consisting of a diuretic, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

[0563] Embodiment 169. A method of increasing plasma non-esterified fatty acids (NEFA) in a patient in need thereof, comprising, consisting essentially of, or consisting of administering to the patient an effective dose of a selectively spliced atrial natriuretic peptide (MANP).

[0564] Embodiment 170. The method of embodiment 169, wherein the increase is an increase in the plasma NEFA from a baseline NEFA by 1 mM within 1 hour after the administration.

[0565] Embodiment 171. The method of embodiment 169 or 170, wherein the MANP is administered subcutaneously.

[0566] Embodiment 172. The method of embodiment 171, wherein the effective dose is 0.1 pg / kg to 5 pg / kg.

[0567] Example 173. The method of Example 172, wherein the effective dose is 2.5 pg / kg.

[0568] Example 174. The method of Example 171, wherein the effective dose is 6.5 pg to 750 pg.

[0569] Example 175. The method of Example 174, wherein the effective dose is 162.5 pg to 375 pg.

[0570] Example 176. The method of Example 169 or 170, wherein the MANP is administered intravenously.

[0571] Example 177. The method of Example 176, wherein the effective dose is 10 pmol / kg / minute to 100 nmol / kg / minute.

[0572] Example 178. The method of Example 169 or 170, wherein the MANP is administered intravenously and subsequently administered subcutaneously.

[0573] Example 179. The method of Example 178, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 0.1 pg / kg to 5 pg / kg.

[0574] Example 180. The method of Example 179, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 2.5 pg / kg.

[0575] Example 181. The method of Example 178, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 6.5 pg to 750 pg.

[0576] Example 182. The method of Example 181, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / minute to 100 nmol / kg / minute and subsequently administered subcutaneously at a dose of 162.5 pg to 375 pg.

[0577] Example 183. The method of any one of Examples 169-182, wherein the patient has a body mass index (BMI) higher than 25 kg / m 2

[0578] ​mg / dL or above prior to the administration. 2

[0579] mg / dL or above prior to the administration. 2

[0580] mg / dL or above prior to the administration.

[0581] mg / dL or above prior to the administration.

[0582] mg / dL or above prior to the administration.

[0583] mg / dL or above prior to the administration.

[0584] mg / dL or above prior to the administration.

[0585] mg / dL or above prior to the administration.

[0586] mg / dL or above prior to the administration.

[0587] ​​Example 193. The method of any one of examples 188-192, wherein the high fasting plasma glucose is identified by a fasting plasma glucose of 100 mg / dL or above in the patient.

[0588] Example 194. The method of any one of examples 188-192, wherein the high fasting plasma glucose is identified in the patient who is receiving pharmacotherapy for elevated blood glucose to lower blood glucose.

[0589] Example 195. The method of any one of examples 169-194, wherein the patient does not present any trait selected from the group consisting of:

[0590] a known hypersensitivity or allergic reaction to the MANP or components thereof, capropil, other natriuretic peptides, or related compounds;

[0591] pregnancy or lactation;

[0592] a history of clinically significant endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal arteries; ingestion of a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration;

[0593] receipt of any investigational drug or device within 30 days prior to the administration;

[0594] a history of alcohol abuse, illicit drug use, severe psychiatric disorder, physiological dependence on any opioid drug, or any history of drug abuse or addiction (within 2 years prior to the administration);

[0595] a history of difficulty donating blood, or donation of blood or blood products within 45 days prior to the administration;

[0596] a clinically significant new illness within 1 month prior to the administration;

[0597] a history of severe allergies;

[0598] a history of coronary artery disease, cerebrovascular disease, or syncope;

[0599] a history of epilepsy or other seizure disorders;

[0600] a history of organ transplantation; and

[0601] a malignancy within 5 years prior to the administration.

[0602] Example 196. The method of any one of examples 169-195, wherein the patient does not present any trait selected from the group consisting of:

[0603] a known hypersensitivity or allergic reaction to the MANP or components thereof, capesantide, other natriuretic peptide or related compound;

[0604] pregnant or nursing;

[0605] has a clinically significant history of endogenous renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery;

[0606] and

[0607] took a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration.

[0608] Embodiment 197. The method of any one of embodiments 169-196, wherein within 12 hours after the administration, the patient’s sitting systolic blood pressure is reduced by 0.1% to 15% compared to the baseline sitting systolic blood pressure.

[0609] Embodiment 198. The method of any one of embodiments 169-197, wherein within 6 hours after the administration, the patient’s sitting diastolic blood pressure is reduced by 0.1% to 5% compared to the baseline sitting diastolic blood pressure.

[0610] Embodiment 199. The method of any one of embodiments 169-198, wherein within 4 hours after the administration, the patient’s plasma glucose is reduced by 1 to 17 mg / mL compared to the baseline plasma glucose.

[0611] Embodiment 200. The method of any one of embodiments 169-199, further comprising, consisting essentially of, or consisting of administering a second medicament selected from the group consisting of a diuretic, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

Claims

1. A method for treating hypertension with metabolic syndrome in a patient requiring this treatment, the patient having a baseline plasma cyclic guanosine monophosphate (cGMP) level below 10 pmol / mL, the method comprising increasing plasma cGMP relative to the baseline level by administering an effective dose of selectively spliced ​​atrial natriuretic peptide (MANP) to the patient.

2. The method according to claim 1, wherein the baseline plasma cGMP is less than 8 pmol / mL.

3. The method according to claim 1 or 2, wherein the increase is to increase the plasma cGMP by 2 to 8 pmol / mL from the baseline within 0.5 hours after the administration.

4. The method according to claim 1 or 2, wherein the increase is to increase the plasma cGMP by 1 to 5 pmol / mL from the baseline within 0.5 to 4 hours after the administration.

5. The method according to any one of claims 1 to 4, wherein the MANP is administered subcutaneously.

6. The method according to claim 5, wherein the effective dose is from 0.1 μg / kg to 5 μg / kg.

7. The method according to claim 6, wherein the effective dose is 2.5 μg / kg.

8. The method according to claim 5, wherein the effective dose is 6.5 μg to 750 μg.

9. The method according to claim 8, wherein the effective dose is from 162.5 μg to 375 μg.

10. The method according to any one of claims 1 to 4, wherein the MANP is administered intravenously.

11. The method of claim 10, wherein the effective dose is from 10 pmol / kg / min to 100 nmol / kg / min.

12. The method according to any one of claims 1 to 4, wherein the MANP is administered intravenously and subsequently subcutaneously.

13. The method of claim 12, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 0.1 μg / kg to 5 μg / kg.

14. The method of claim 13, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 2.5 μg / kg.

15. The method of claim 12, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 6.5 μg to 750 μg.

16. The method of claim 15, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 162.5 μg to 375 μg.

17. The method according to any one of claims 1 to 16, wherein the patient has a weight of more than 25 kg / m². 2 Body Mass Index (BMI).

18. The method according to any one of claims 1 to 17, wherein the patient has a weight greater than 30 kg / m². 2 BMI.

19. The method according to any one of claims 1 to 18, wherein the patient has a weight greater than 35 kg / m². 2 BMI.

20. The method according to any one of claims 1 to 19, wherein the patient presents at least two traits selected from the group consisting of: abdominal obesity, high serum triglycerides, low serum high-density lipoprotein (HDL) cholesterol, and high fasting plasma glucose.

21. The method of claim 20, wherein the abdominal obesity is identified by a waist circumference greater than or equal to 102 cm for the male patient or greater than or equal to 88 cm for the female patient.

22. The method of claim 20 or 21, wherein the high serum triglycerides are identified by serum triglycerides of 150 mg / dL or higher in the patient prior to the administration.

23. The method according to any one of claims 20 to 22, wherein the low HDL cholesterol is identified by HDL cholesterol levels below 40 mg / dL in the male patient or below 50 mg / dL in the female patient.

24. The method according to any one of claims 20 to 22, wherein the low HDL cholesterol is identified in the patient who is receiving drug treatment for low HDL cholesterol to increase HDL cholesterol.

25. The method according to any one of claims 20 to 24, wherein the high fasting plasma glucose is identified by fasting plasma glucose of 100 mg / dL or higher in the patient.

26. The method according to any one of claims 20 to 24, wherein the high fasting plasma glucose is identified in the patient who is receiving drug treatment for elevated blood glucose to lower blood glucose.

27. The method according to any one of claims 1 to 26, wherein the patient has a sitting systolic blood pressure between 140 mmHg and 180 mmHg or a sitting diastolic blood pressure between 90 mmHg and 100 mmHg prior to the administration.

28. The method according to any one of claims 1 to 27, wherein the patient has taken an antihypertensive drug for 30 days prior to the administration, or has taken a statin, ezetimibe, or a combination thereof for 60 days prior to the administration.

29. The method according to any one of claims 1 to 28, wherein the patient does not exhibit any trait selected from the group consisting of: Known hypersensitivity or allergic reaction to the aforementioned MANP or its components, capperidin, other diuretic natriuretic peptides or related compounds; Being pregnant or breastfeeding; A history of clinically significant endogenous nephropathy, renal artery stenosis, or renal artery fibromuscular dysplasia; Take a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration; Has received any investigational drug or device within 30 days prior to administration; (Within 2 years prior to the administration) a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or any drug abuse or addiction; Those with a history of difficulty donating blood, or who have donated blood or blood products within 45 days prior to the administration of the product; Having a clinically significant new disease within one month prior to the application; History of severe allergies; A history of coronary artery disease, cerebrovascular disease, or syncope; A history of epilepsy or other epileptic seizures; History of organ transplantation; and The patient had a malignant tumor within 5 years prior to the application.

30. The method according to any one of claims 1 to 29, wherein the patient does not exhibit any trait selected from the group consisting of: Known hypersensitivity or allergic reaction to the aforementioned MANP or its components, capperidin, other diuretic natriuretic peptides or related compounds; Being pregnant or breastfeeding; A history of clinically significant endogenous nephropathy, renal artery stenosis, or renal artery fibromuscular dysplasia; and Take a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration.

31. The method according to any one of claims 1 to 30, wherein within 12 hours after the administration, the patient's sitting systolic blood pressure decreases by 0.1% to 15% compared to baseline sitting systolic blood pressure.

32. The method according to any one of claims 1 to 31, wherein within 6 hours after the administration, the patient's sitting diastolic blood pressure decreases by 0.1% to 5% compared to baseline sitting diastolic blood pressure.

33. The method according to any one of claims 1 to 32, wherein within 4 hours after the administration, the patient's plasma glucose decreases by 1 to 17 mg / mL compared to baseline plasma glucose.

34. The method according to any one of claims 1 to 33, further comprising administering a second drug selected from the group consisting of a diuretic, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

35. A method for lowering blood pressure in a patient with a baseline plasma cyclic guanosine monophosphate (cGMP) of less than 10 pmol / mL, the method comprising increasing plasma cGMP relative to the baseline by administering an effective dose of selectively spliced ​​atrial natriuretic peptide (MANP) to the patient.

36. The method of claim 35, wherein the baseline plasma cGMP is less than 8 pmol / mL.

37. The method of claim 35 or 36, wherein the increase is to increase the plasma cGMP by 2 to 8 pmol / mL from the baseline within 0.5 hours after the administration.

38. The method of claim 35 or 36, wherein the increase is to increase the plasma cGMP by 1 to 5 pmol / mL from the baseline within 0.5 to 4 hours after the administration.

39. The method according to any one of claims 35 to 38, wherein the reduction is a reduction of the patient's sitting systolic blood pressure by 0.1% to 15% compared to baseline sitting systolic blood pressure within 12 hours after the administration.

40. The method according to any one of claims 35 to 38, wherein the reduction is a reduction of 0.1% to 5% in the patient's sitting diastolic blood pressure compared to baseline sitting diastolic blood pressure within 6 hours after the administration.

41. The method according to any one of claims 35 to 40, wherein the MANP is administered subcutaneously.

42. The method of claim 41, wherein the effective dose is from 0.1 μg / kg to 5 μg / kg.

43. The method according to claim 42, wherein the effective dose is 2.5 μg / kg.

44. The method of claim 41, wherein the effective dose is 6.5 μg to 750 μg.

45. The method of claim 44, wherein the effective dose is from 162.5 μg to 375 μg.

46. ​​The method according to any one of claims 35 to 40, wherein the MANP is administered intravenously.

47. The method of claim 46, wherein the effective dose is from 10 pmol / kg / min to 100 nmol / kg / min.

48. The method according to any one of claims 35 to 40, wherein the MANP is administered intravenously and subsequently subcutaneously.

49. The method of claim 48, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 0.1 μg / kg to 5 μg / kg.

50. The method of claim 49, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 2.5 μg / kg.

51. The method of claim 48, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 6.5 μg to 750 μg.

52. The method of claim 51, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 162.5 μg to 375 μg.

53. The method according to any one of claims 35 to 52, wherein the patient has a body temperature above 25 kg / m². 2 Body Mass Index (BMI).

54. The method according to any one of claims 35 to 53, wherein the patient has a weight greater than 30 kg / m². 2 BMI.

55. The method according to any one of claims 35 to 54, wherein the patient has a body temperature above 35 kg / m². 2 BMI.

56. The method according to any one of claims 35 to 55, wherein the patient presents a sitting systolic blood pressure between 140 mmHg and 180 mmHg or a sitting diastolic blood pressure between 90 mmHg and 100 mmHg prior to the administration.

57. The method according to any one of claims 35 to 56, wherein the patient has taken an antihypertensive drug for 30 days prior to the administration, or has taken a statin, ezetimibe, or a combination thereof for 60 days prior to the administration.

58. The method according to any one of claims 35 to 57, wherein the patient presents at least two characteristics selected from the group consisting of: abdominal obesity, high serum triglycerides, low serum high-density lipoprotein (HDL) cholesterol, and high fasting plasma glucose.

59. The method of claim 58, wherein the abdominal obesity is identified by a waist circumference greater than or equal to 102 cm for the male patient or greater than or equal to 88 cm for the female patient.

60. The method according to claim 58 or 59, wherein the high serum triglycerides are identified by serum triglycerides of 150 mg / dL or higher in the patient prior to the administration.

61. The method according to any one of claims 58 to 60, wherein the low HDL cholesterol is identified by HDL cholesterol levels below 40 mg / dL in the male patient or below 50 mg / dL in the female patient.

62. The method according to any one of claims 58 to 60, wherein the low HDL cholesterol is identified in the patient who is receiving drug treatment for low HDL cholesterol to increase HDL cholesterol.

63. The method according to any one of claims 58 to 62, wherein the high fasting plasma glucose is identified by fasting plasma glucose of 100 mg / dL or higher in the patient.

64. The method according to any one of claims 58 to 62, wherein the high fasting plasma glucose is identified in the patient who is receiving drug treatment for elevated blood glucose to lower blood glucose.

65. The method according to any one of claims 35 to 64, wherein the patient does not exhibit any trait selected from the group consisting of: Known hypersensitivity or allergic reaction to the aforementioned MANP or its components, capperidin, other diuretic natriuretic peptides or related compounds; Being pregnant or breastfeeding; A history of clinically significant endogenous nephropathy, renal artery stenosis, or renal artery fibromuscular dysplasia; Take a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration; Has received any investigational drug or device within 30 days prior to administration; (Within 2 years prior to the administration) a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or any drug abuse or addiction; Those with a history of difficulty donating blood, or who have donated blood or blood products within 45 days prior to the administration of the product; Having a clinically significant new disease within one month prior to the application; History of severe allergies; A history of coronary artery disease, cerebrovascular disease, or syncope; A history of epilepsy or other epileptic seizures; History of organ transplantation; and The patient had a malignant tumor within 5 years prior to the application.

66. The method according to any one of claims 35 to 65, wherein the patient does not exhibit any trait selected from the group consisting of: Known hypersensitivity or allergic reaction to the aforementioned MANP or its components, capperidin, other diuretic natriuretic peptides or related compounds; Being pregnant or breastfeeding; A history of clinically significant endogenous nephropathy, renal artery stenosis, or renal artery fibromuscular dysplasia; and Take a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration.

67. The method according to any one of claims 35 to 66, wherein within 4 hours after the administration, the patient's plasma glucose decreases by 1 to 17 mg / mL compared to baseline plasma glucose.

68. The method according to any one of claims 35 to 67, further comprising administering a second drug selected from the group consisting of a diuretic, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

69. A method for increasing plasma atrial natriuretic peptide (ANP)-like peptide in a patient with a baseline plasma cyclic guanosine monophosphate (cGMP) of less than 10 pmol / mL, the method comprising increasing plasma cGMP relative to the baseline by administering an effective dose of selectively spliced ​​atrial natriuretic peptide (MANP) to the patient.

70. The method of claim 69, wherein the baseline plasma cGMP is less than 8 pmol / mL.

71. The method of claim 69 or 70, wherein the increase in plasma cGMP is achieved by increasing plasma cGMP from baseline by 2 to 8 pmol / mL within 0.5 hours after administration.

72. The method of claim 69 or 70, wherein the increase in plasma cGMP is achieved by increasing plasma cGMP by 1 to 5 pmol / mL from baseline within 0.5 to 4 hours after administration.

73. The method according to any one of claims 69 to 72, wherein the increase in ANP-like peptide is to increase the ANP-like peptide from baseline ANP-like peptide by 1 to 70 pg / mL within 0.5 hours after the administration.

74. The method according to any one of claims 69 to 72, wherein the increase in ANP-like peptide is to increase the ANP-like peptide from baseline ANP-like peptide by 1 to 40 pg / mL within 1 hour after the administration.

75. The method according to any one of claims 69 to 74, wherein the MANP is administered subcutaneously.

76. The method of claim 75, wherein the effective dose is from 0.1 μg / kg to 5 μg / kg.

77. The method of claim 76, wherein the effective dose is 2.5 μg / kg.

78. The method of claim 75, wherein the effective dose is from 6.5 μg to 750 μg.

79. The method of claim 78, wherein the effective dose is from 162.5 μg to 375 μg.

80. The method according to any one of claims 69 to 74, wherein the MANP is administered intravenously.

81. The method of claim 80, wherein the effective dose is from 10 pmol / kg / min to 100 nmol / kg / min.

82. The method according to any one of claims 69 to 74, wherein the MANP is administered intravenously and subsequently subcutaneously.

83. The method of claim 82, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 0.1 μg / kg to 5 μg / kg.

84. The method of claim 83, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 2.5 μg / kg.

85. The method of claim 82, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 6.5 μg to 750 μg.

86. The method of claim 85, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 162.5 μg to 375 μg.

87. The method according to any one of claims 69 to 86, wherein the patient has a weight greater than 25 kg / m². 2 Body Mass Index (BMI).

88. The method according to any one of claims 69 to 87, wherein the patient has a weight greater than 30 kg / m². 2 BMI.

89. The method according to any one of claims 69 to 88, wherein the patient has a weight greater than 35 kg / m². 2 BMI.

90. The method according to any one of claims 69 to 89, wherein the patient presents a sitting systolic blood pressure between 140 mmHg and 180 mmHg or a sitting diastolic blood pressure between 90 mmHg and 100 mmHg prior to the administration.

91. The method according to any one of claims 69 to 90, wherein the patient has taken an antihypertensive drug for 30 days prior to the administration, or has taken a statin, ezetimibe, or a combination thereof for 60 days prior to the administration.

92. The method according to any one of claims 69 to 91, wherein the patient presents at least two traits selected from the group consisting of: abdominal obesity, high serum triglycerides, low serum high-density lipoprotein (HDL) cholesterol, and high fasting plasma glucose.

93. The method of claim 92, wherein the abdominal obesity is identified by a waist circumference greater than or equal to 102 cm for the male patient or greater than or equal to 88 cm for the female patient.

94. The method according to claim 92 or 93, wherein the high serum triglycerides are identified by serum triglycerides of 150 mg / dL or higher in the patient prior to the administration.

95. The method according to any one of claims 92 to 94, wherein the low HDL cholesterol is identified by HDL cholesterol levels below 40 mg / dL in the male patient or below 50 mg / dL in the female patient.

96. The method according to any one of claims 92 to 94, wherein the low HDL cholesterol is identified in the patient who is receiving drug treatment for low HDL cholesterol to increase HDL cholesterol.

97. The method according to any one of claims 92 to 96, wherein the high fasting plasma glucose is identified by fasting plasma glucose of 100 mg / dL or higher in the patient.

98. The method according to any one of claims 92 to 96, wherein the high fasting plasma glucose is identified in the patient who is receiving drug treatment for elevated blood glucose to lower blood glucose.

99. The method according to any one of claims 69 to 98, wherein the patient does not exhibit any trait selected from the group consisting of: Known hypersensitivity or allergic reaction to the aforementioned MANP or its components, capperidin, other diuretic natriuretic peptides or related compounds; Being pregnant or breastfeeding; A history of clinically significant endogenous nephropathy, renal artery stenosis, or renal artery fibromuscular dysplasia; Take a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration; Has received any investigational drug or device within 30 days prior to administration; (Within 2 years prior to the administration) a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or any drug abuse or addiction; Those with a history of difficulty donating blood, or who have donated blood or blood products within 45 days prior to the administration of the product; Having a clinically significant new disease within one month prior to the application; History of severe allergies; A history of coronary artery disease, cerebrovascular disease, or syncope; A history of epilepsy or other epileptic seizures; History of organ transplantation; and The patient had a malignant tumor within 5 years prior to the application.

100. The method according to any one of claims 69 to 99, wherein the patient does not exhibit any trait selected from the group consisting of: Known hypersensitivity or allergic reaction to the aforementioned MANP or its components, capperidin, other diuretic natriuretic peptides or related compounds; Being pregnant or breastfeeding; A history of clinically significant endogenous nephropathy, renal artery stenosis, or renal artery fibromuscular dysplasia; and Take a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration.

101. The method according to any one of claims 69 to 100, wherein within 12 hours after the administration, the patient's sitting systolic blood pressure decreases by 0.1% to 15% compared to baseline sitting systolic blood pressure.

102. The method according to any one of claims 69 to 101, wherein within 6 hours after the administration, the patient's sitting diastolic blood pressure decreases by 0.1% to 5% compared to baseline sitting diastolic blood pressure.

103. The method according to any one of claims 69 to 102, wherein within 4 hours after the administration, the patient's plasma glucose decreases by 1 to 17 mg / mL compared to baseline plasma glucose.

104. The method according to any one of claims 69 to 103, further comprising administering a second drug selected from the group consisting of a diuretic, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

105. A method for reducing plasma glucose levels in a patient who requires this, comprising administering to the patient an effective dose of selectively spliced ​​atrial natriuretic peptide (MANP).

106. The method of claim 105, wherein the reduction is to reduce the plasma glucose level by 1 to 17 mg / mL compared to baseline plasma glucose within 4 hours after the administration.

107. The method of claim 105 or 106, wherein the MANP is administered subcutaneously.

108. The method of claim 107, wherein the effective dose is from 0.1 μg / kg to 5 μg / kg.

109. The method according to claim 108, wherein the effective dose is 2.5 μg / kg.

110. The method of claim 107, wherein the effective dose is 6.5 μg to 750 μg.

111. The method of claim 110, wherein the effective dose is from 162.5 μg to 375 μg.

112. The method of claim 105 or 106, wherein the MANP is administered intravenously.

113. The method of claim 112, wherein the effective dose is from 10 pmol / kg / min to 100 nmol / kg / min.

114. The method of claim 105 or 106, wherein the MANP is administered intravenously and subsequently subcutaneously.

115. The method of claim 114, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 0.1 μg / kg to 5 μg / kg.

116. The method of claim 115, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 2.5 μg / kg.

117. The method of claim 114, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 6.5 μg to 750 μg.

118. The method of claim 117, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 162.5 μg to 375 μg.

119. The method according to any one of claims 105 to 118, wherein the patient has a weight of more than 25 kg / m². 2 Body Mass Index (BMI).

120. The method according to any one of claims 105 to 119, wherein the patient has a weight of more than 30 kg / m². 2 BMI.

121. The method according to any one of claims 105 to 120, wherein the patient has a weight of more than 35 kg / m². 2 BMI.

122. The method according to any one of claims 105 to 121, wherein the patient presents a sitting systolic blood pressure between 140 mmHg and 180 mmHg or a sitting diastolic blood pressure between 90 mmHg and 100 mmHg prior to the administration.

123. The method according to any one of claims 105 to 122, wherein the patient has taken an antihypertensive drug for 30 days prior to the administration, or has taken a statin, ezetimibe, or a combination thereof for 60 days prior to the administration.

124. The method according to any one of claims 105 to 123, wherein the patient presents at least two traits selected from the group consisting of: abdominal obesity, high serum triglycerides, low serum high-density lipoprotein (HDL) cholesterol, and high fasting plasma glucose.

125. The method of claim 124, wherein the abdominal obesity is identified by a waist circumference greater than or equal to 102 cm for the male patient or greater than or equal to 88 cm for the female patient.

126. The method of claim 124 or 125, wherein the high serum triglycerides are identified by serum triglycerides of 150 mg / dL or higher in the patient prior to the administration.

127. The method according to any one of claims 124 to 126, wherein the low HDL cholesterol is identified by HDL cholesterol levels below 40 mg / dL in the male patient or below 50 mg / dL in the female patient.

128. The method according to any one of claims 124 to 126, wherein the low HDL cholesterol is identified in the patient who is receiving drug treatment for low HDL cholesterol to increase HDL cholesterol.

129. The method according to any one of claims 124 to 128, wherein the high fasting plasma glucose is identified by fasting plasma glucose of 100 mg / dL or higher in the patient.

130. The method according to any one of claims 124 to 128, wherein the high fasting plasma glucose is identified in the patient who is receiving drug treatment for elevated blood glucose to lower blood glucose.

131. The method according to any one of claims 105 to 130, wherein the patient does not exhibit any trait selected from the group consisting of: Known hypersensitivity or allergic reaction to the aforementioned MANP or its components, capperidin, other diuretic natriuretic peptides or related compounds; Being pregnant or breastfeeding; A history of clinically significant endogenous nephropathy, renal artery stenosis, or renal artery fibromuscular dysplasia; Take a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration; Has received any investigational drug or device within 30 days prior to administration; (Within 2 years prior to the administration) a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or any drug abuse or addiction; Those with a history of difficulty donating blood, or who have donated blood or blood products within 45 days prior to the administration of the product; Having a clinically significant new disease within one month prior to the application; History of severe allergies; A history of coronary artery disease, cerebrovascular disease, or syncope; A history of epilepsy or other epileptic seizures; History of organ transplantation; and The patient had a malignant tumor within 5 years prior to the application.

132. The method according to any one of claims 105 to 131, wherein the patient does not exhibit any trait selected from the group consisting of: Known hypersensitivity or allergic reaction to the aforementioned MANP or its components, capperidin, other diuretic natriuretic peptides or related compounds; Being pregnant or breastfeeding; A history of clinically significant endogenous nephropathy, renal artery stenosis, or renal artery fibromuscular dysplasia; and Take a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration.

133. The method according to any one of claims 105 to 132, wherein within 12 hours after the administration, the patient's sitting systolic blood pressure decreases by 0.1% to 15% compared to baseline sitting systolic blood pressure.

134. The method according to any one of claims 105 to 133, wherein within 6 hours after the administration, the patient's sitting diastolic blood pressure decreases by 0.1% to 5% compared to baseline sitting diastolic blood pressure.

135. The method according to any one of claims 105 to 134, further comprising administering a second drug selected from the group consisting of a diuretic, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

136. A method for increasing insulin sensitivity in a patient with this need, comprising administering to the patient an effective dose of selectively spliced ​​atrial natriuretic peptide (MANP).

137. The method of claim 136, wherein the increase in insulin sensitivity is achieved by increasing the patient's updated homeostatic model assessment model insulin sensitivity (HOMA2-S) value by 0.1% to 35% compared to the baseline HOMA2-S value within 4 hours after administration.

138. The method of claim 136, wherein the increase in insulin sensitivity is achieved by reducing the patient's updated homeostatic model assessment model insulin resistance (HOMA2-IR) value by 0.1 to 0.6 from the baseline HOMA2-IR value within 4 hours after administration.

139. The method according to any one of claims 136 to 138, wherein the MANP is administered subcutaneously.

140. The method of claim 139, wherein the effective dose is from 0.1 μg / kg to 5 μg / kg.

141. The method of claim 140, wherein the effective dose is 2.5 μg / kg.

142. The method of claim 139, wherein the effective dose is 6.5 μg to 750 μg.

143. The method of claim 142, wherein the effective dose is from 162.5 μg to 375 μg.

144. The method according to any one of claims 136 to 138, wherein the MANP is administered intravenously.

145. The method of claim 144, wherein the effective dose is from 10 pmol / kg / min to 100 nmol / kg / min.

146. The method according to any one of claims 136 to 138, wherein the MANP is administered intravenously and subsequently subcutaneously.

147. The method of claim 146, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 0.1 μg / kg to 5 μg / kg.

148. The method of claim 147, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 2.5 μg / kg.

149. The method of claim 146, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 6.5 μg to 750 μg.

150. The method of claim 149, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 162.5 μg to 375 μg.

151. The method according to any one of claims 136 to 150, wherein the patient has a weight of more than 25 kg / m². 2 Body Mass Index (BMI).

152. The method according to any one of claims 136 to 151, wherein the patient has a weight greater than 30 kg / m². 2 BMI.

153. The method according to any one of claims 136 to 152, wherein the patient has a weight of more than 35 kg / m². 2 BMI.

154. The method according to any one of claims 136 to 153, wherein the patient presents a sitting systolic blood pressure between 140 mmHg and 180 mmHg or a sitting diastolic blood pressure between 90 mmHg and 100 mmHg prior to the administration.

155. The method according to any one of claims 136 to 154, wherein the patient has taken an antihypertensive drug for 30 days prior to the administration, or has taken a statin, ezetimibe, or a combination thereof for 60 days prior to the administration.

156. The method according to any one of claims 136 to 155, wherein the patient presents at least two traits selected from the group consisting of: abdominal obesity, high serum triglycerides, low serum high-density lipoprotein (HDL) cholesterol, and high fasting plasma glucose.

157. The method of claim 156, wherein the abdominal obesity is identified by a waist circumference greater than or equal to 102 cm for the male patient or greater than or equal to 88 cm for the female patient.

158. The method according to claim 156 or 157, wherein the high serum triglycerides are identified by serum triglycerides of 150 mg / dL or higher in the patient prior to the administration.

159. The method according to any one of claims 156 to 158, wherein the low HDL cholesterol is identified by HDL cholesterol levels below 40 mg / dL in the male patient or below 50 mg / dL in the female patient.

160. The method according to any one of claims 156 to 158, wherein the low HDL cholesterol is identified in the patient who is receiving drug treatment for low HDL cholesterol to increase HDL cholesterol.

161. The method according to any one of claims 156 to 160, wherein the high fasting plasma glucose is identified by fasting plasma glucose of 100 mg / dL or more in the patient.

162. The method according to any one of claims 156 to 160, wherein the high fasting plasma glucose is identified in the patient who is receiving drug treatment for elevated blood glucose to lower blood glucose.

163. The method according to any one of claims 136 to 162, wherein the patient does not exhibit any trait selected from the group consisting of: Known hypersensitivity or allergic reaction to the aforementioned MANP or its components, capperidin, other diuretic natriuretic peptides or related compounds; Being pregnant or breastfeeding; A history of clinically significant endogenous nephropathy, renal artery stenosis, or renal artery fibromuscular dysplasia; Take a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration; Has received any investigational drug or device within 30 days prior to administration; (Within 2 years prior to the administration) a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or any drug abuse or addiction; Those with a history of difficulty donating blood, or who have donated blood or blood products within 45 days prior to the administration of the product; Having a clinically significant new disease within one month prior to the application; History of severe allergies; A history of coronary artery disease, cerebrovascular disease, or syncope; A history of epilepsy or other epileptic seizures; History of organ transplantation; and The patient had a malignant tumor within 5 years prior to the application.

164. The method according to any one of claims 136 to 163, wherein the patient does not exhibit any trait selected from the group consisting of: Known hypersensitivity or allergic reaction to the aforementioned MANP or its components, capperidin, other diuretic natriuretic peptides or related compounds; Being pregnant or breastfeeding; A history of clinically significant endogenous nephropathy, renal artery stenosis, or renal artery fibromuscular dysplasia; and Take a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration.

165. The method according to any one of claims 136 to 164, wherein within 12 hours after the administration, the patient's sitting systolic blood pressure decreases by 0.1% to 15% compared to baseline sitting systolic blood pressure.

166. The method according to any one of claims 136 to 165, wherein within 6 hours after the administration, the patient's sitting diastolic blood pressure decreases by 0.1% to 5% compared to baseline sitting diastolic blood pressure.

167. The method according to any one of claims 136 to 166, wherein within 4 hours after the administration, the patient's plasma glucose decreases by 1 to 17 mg / mL compared to baseline plasma glucose.

168. The method according to any one of claims 136 to 167, further comprising administering a second drug selected from the group consisting of a diuretic, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

169. A method for increasing plasma non-esterified fatty acids (NEFA) in a patient with this need, comprising administering to said patient an effective dose of selectively spliced ​​atrial natriuretic peptide (MANP).

170. The method of claim 169, wherein the increase is to increase the plasma NEFA by 1 μM from baseline NEFA within 1 hour after the administration.

171. The method of claim 169 or 170, wherein the MANP is administered subcutaneously.

172. The method of claim 171, wherein the effective dose is from 0.1 μg / kg to 5 μg / kg.

173. The method according to claim 172, wherein the effective dose is 2.5 μg / kg.

174. The method of claim 171, wherein the effective dose is 6.5 μg to 750 μg.

175. The method of claim 174, wherein the effective dose is from 162.5 μg to 375 μg.

176. The method of claim 169 or 170, wherein the MANP is administered intravenously.

177. The method of claim 176, wherein the effective dose is from 10 pmol / kg / min to 100 nmol / kg / min.

178. The method of claim 169 or 170, wherein the MANP is administered intravenously and subsequently subcutaneously.

179. The method of claim 178, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 0.1 μg / kg to 5 μg / kg.

180. The method of claim 179, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 2.5 μg / kg.

181. The method of claim 178, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 6.5 μg to 750 μg.

182. The method of claim 181, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min and subsequently subcutaneously at a dose of 162.5 μg to 375 μg.

183. The method according to any one of claims 169 to 182, wherein the patient has a weight of more than 25 kg / m². 2 Body Mass Index (BMI).

184. The method according to any one of claims 169 to 183, wherein the patient has a weight greater than 30 kg / m². 2 BMI.

185. The method according to any one of claims 169 to 184, wherein the patient has a weight of more than 35 kg / m². 2 BMI.

186. The method according to any one of claims 169 to 185, wherein the patient presents a sitting systolic blood pressure between 140 mmHg and 180 mmHg or a sitting diastolic blood pressure between 90 mmHg and 100 mmHg prior to the administration.

187. The method according to any one of claims 169 to 186, wherein the patient has taken an antihypertensive drug for 30 days prior to the administration, or has taken a statin, ezetimibe, or a combination thereof for 60 days prior to the administration.

188. The method according to any one of claims 169 to 187, wherein the patient presents at least two traits selected from the group consisting of: abdominal obesity, high serum triglycerides, low serum high-density lipoprotein (HDL) cholesterol, and high fasting plasma glucose.

189. The method of claim 188, wherein the abdominal obesity is identified by a waist circumference greater than or equal to 102 cm for the male patient or greater than or equal to 88 cm for the female patient.

190. The method according to claim 188 or 189, wherein the high serum triglycerides are identified by serum triglycerides of 150 mg / dL or higher in the patient prior to the administration.

191. The method according to any one of claims 188 to 190, wherein the low HDL cholesterol is identified by HDL cholesterol levels below 40 mg / dL in the male patient or below 50 mg / dL in the female patient.

192. The method according to any one of claims 188 to 190, wherein the low HDL cholesterol is identified in the patient who is receiving drug treatment for low HDL cholesterol to increase HDL cholesterol.

193. The method according to any one of claims 188 to 192, wherein the high fasting plasma glucose is identified by fasting plasma glucose of 100 mg / dL or higher in the patient.

194. The method according to any one of claims 188 to 192, wherein the high fasting plasma glucose is identified in the patient who is receiving drug treatment for elevated blood glucose to lower blood glucose.

195. The method according to any one of claims 169 to 194, wherein the patient does not exhibit any trait selected from the group consisting of: Known hypersensitivity or allergic reaction to the aforementioned MANP or its components, capperidin, other diuretic natriuretic peptides or related compounds; Being pregnant or breastfeeding; A history of clinically significant endogenous nephropathy, renal artery stenosis, or renal artery fibromuscular dysplasia; Take a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration; Has received any investigational drug or device within 30 days prior to administration; (Within 2 years prior to the administration) a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or any drug abuse or addiction; Those with a history of difficulty donating blood, or who have donated blood or blood products within 45 days prior to the administration of the product; Having a clinically significant new disease within one month prior to the application; History of severe allergies; A history of coronary artery disease, cerebrovascular disease, or syncope; A history of epilepsy or other epileptic seizures; History of organ transplantation; and The patient had a malignant tumor within 5 years prior to the application.

196. The method according to any one of claims 169 to 195, wherein the patient does not exhibit any trait selected from the group consisting of: Known hypersensitivity or allergic reaction to the aforementioned MANP or its components, capperidin, other diuretic natriuretic peptides or related compounds; Being pregnant or breastfeeding; A history of clinically significant endogenous nephropathy, renal artery stenosis, or renal artery fibromuscular dysplasia; and Take a phosphodiesterase type 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administration.

197. The method according to any one of claims 169 to 196, wherein within 12 hours after the administration, the patient's sitting systolic blood pressure decreases by 0.1% to 15% compared to baseline sitting systolic blood pressure.

198. The method according to any one of claims 169 to 197, wherein within 6 hours after the administration, the patient's sitting diastolic blood pressure decreases by 0.1% to 5% compared to baseline sitting diastolic blood pressure.

199. The method according to any one of claims 169 to 198, wherein within 4 hours after the administration, the patient's plasma glucose decreases by 1 to 17 mg / mL compared to baseline plasma glucose.

200. The method according to any one of claims 169 to 199, further comprising administering a second drug selected from the group consisting of a diuretic, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

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