Treatment method for hypertension with metabolic syndrome using MANP

Abstract

The present disclosure provides methods and materials related to MANPs. In one embodiment, the present disclosure provides a method for treating hypertension with metabolic syndrome by administering MANPs to a patient in need of such treatment. In one embodiment, the present disclosure provides a method comprising administering MANPs to a patient to lower blood pressure (BP). In one embodiment, the method of the present disclosure is employed to treat patients with low baseline cGMP levels. In one embodiment, the present disclosure provides a therapeutic method using MANPs to increase or improve plasma cGMP levels, plasma ANP-like peptide levels, insulin sensitivity, or plasma NEFA levels. In one embodiment, the present disclosure provides a therapeutic method using MANPs to lower plasma glucose levels.

Description

[Technical Field]

[0001] Related Applications This application claims priority to U.S. Provisional Application No. 63 / 486,421, filed February 22, 2023, the entire contents of which are incorporated herein by reference.

[0002] Statement Regarding Federally Sponsored Research This invention was made with government support under HL136340 and HL134668 awarded by the National Institutes of Health. The government has certain rights in this invention.

[0003] The present disclosure provides methods of treating hypertension with metabolic syndrome in patients using alternatively spliced ​​atrial natriuretic peptide (MANP). [Background technology]

[0004] Hypertension, also known as high blood pressure (BP), is a long-term condition characterized by persistently elevated arterial blood pressure (BP). Metabolic syndrome is a constellation of cardiovascular and metabolic risk factors that predispose patients to significant cardiovascular disease. Hypertension is one of the core clinical features of metabolic syndrome. In fact, 77% of patients with metabolic syndrome also have hypertension, and vice versa. Visceral obesity is a significant risk factor for the development of hypertension. While hypertension and metabolic syndrome are each significant risk factors for cardiovascular disease alone, their coexistence doubles the risk. Despite the two interrelated conditions, none of the currently available antihypertensive drugs have been reported to exert favorable metabolic effects, and there are no therapies specifically approved for the treatment of hypertension associated with metabolic syndrome.

[0005] The difficulty in treating hypertension with metabolic syndrome may be due to the variable response of each patient to potential treatments. The lack of objective and reliable predictors of treatment outcome is one of the many reasons why there are no approved medications for the treatment of hypertension with metabolic syndrome. While published studies (see, for example, Chen et al. (2021), Hypertension, 78(6):1859-1867) have shown that MANP may be administered to regulate certain symptoms of resistant hypertension, there are currently no known predictors that can reliably predict a patient's response to any hypertension treatment for the treatment of hypertension with metabolic syndrome. To provide effective treatment for patients with hypertension with metabolic syndrome, treatments using reliable predictors remain necessary. Summary of the Invention

[0006] The present disclosure includes methods and materials related to MANPs. The disclosure also includes a method for treating hypertension with metabolic syndrome in a patient in need of such treatment who has low baseline plasma cyclic guanosine monophosphate (cGMP) by administering MANPs to the patient. In one embodiment, the disclosure includes administering MANPs to a patient in need of such treatment to increase plasma cGMP levels, lower blood pressure, or increase plasma atrial natriuretic peptide (ANP)-like peptide levels in the patient. In one embodiment, the disclosure also includes administering MANPs to a patient to lower plasma glucose levels, improve insulin sensitivity, or increase plasma nonesterified fatty acid (NEFA) levels.

[0007] In one embodiment, the present disclosure provides and includes a method for treating hypertension with metabolic syndrome, the method being applied to a patient in need of such treatment and having a baseline plasma cGMP level below 10 pmol / mL. The method of the present disclosure comprises administering to the patient an effective amount of MANP to increase the plasma cGMP level relative to baseline. In one embodiment, the present disclosure provides and includes a method for lowering blood pressure, the method being applied to a patient in need of such treatment and having a baseline plasma cGMP level below 10 pmol / mL. The method of the present disclosure comprises administering to the patient an effective amount of MANP to increase the plasma cGMP level relative to baseline. In one embodiment, the present disclosure provides and includes a method for increasing plasma ANP-like peptide levels, the method being applied to a patient in need of such treatment and having a baseline plasma cGMP level below 10 pmol / mL. The method of the present disclosure comprises administering to the patient an effective amount of MANP to increase the plasma cGMP level relative to baseline. In one aspect, the present disclosure provides and includes a method for lowering plasma glucose levels, the method being administered to a patient in need of such treatment and comprising administering to the patient an effective amount of MANP. In one aspect, the present disclosure provides and includes a method for improving insulin sensitivity, the method being administered to a patient in need of such treatment and comprising administering to the patient an effective amount of MANP. In one aspect, the present disclosure provides and includes a method for increasing plasma NEFA levels, the method being administered to a patient in need of such treatment and comprising administering to the patient an effective amount of MANP.

[0008] In one embodiment, the method of the present disclosure increases plasma cGMP levels by 2 to 8 pmol / mL from baseline within 0.5 hours after administration of an effective amount of MANP in a patient in need of such treatment. In one embodiment, the method of lowering blood pressure, administered to a patient in need of such treatment, reduces sitting systolic blood pressure by 0.1 to 15% compared to baseline sitting systolic blood pressure within 12 hours after administration of an effective amount of MANP. In one embodiment, the method of lowering blood pressure, administered to a patient in need of such treatment, reduces sitting diastolic blood pressure by 0.1 to 5% compared to baseline sitting diastolic blood pressure within 6 hours after administration of an effective amount of MANP. In one embodiment, the method of increasing plasma ANP-like peptide levels, administered to a patient in need of such treatment, increases plasma ANP-like peptide levels by 1 to 70 pg / mL compared to baseline plasma ANP-like peptide levels within 0.5 hours after administration of an effective amount of MANP. In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment increases the patient's updated homeostasis model assessment sensitivity (HOMA2-S) value by 0.1 to 35% relative to the patient's baseline HOMA2-S value within 4 hours after administration of an effective amount of MANP. In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment decreases the patient's HOMA2 resistance (HOMA2-IR) value by 0.1 to 0.6 relative to the patient's baseline HOMA2-IR value within 4 hours after administration of an effective amount of MANP. In one embodiment, a method for increasing plasma NEFA levels administered to a patient in need of such treatment increases plasma NEFA levels by 1 μM relative to the patient's baseline plasma NEFA level within 1 hour after administration of an effective amount of MANP.

[0009] In one embodiment, 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 one embodiment, MANP can be administered intravenously (e.g., at a dose of about 10 pmol / kg / min to 100 nmol / kg / min). In one embodiment, MANP can be administered intravenously followed by subcutaneous administration. For example, MANP can be administered intravenously at a dose of about 10 pmol / kg / min to about 100 nmol / kg / min, followed by subcutaneous administration at a dose of about 0.1 μg / kg to 5 μg / kg.

[0010] In one embodiment, the patient in need of such treatment exhibits hypertension despite the use of at least one antihypertensive medication. In one embodiment, the patient in need of such treatment has a sitting systolic blood pressure of 140 mmHg to 180 mmHg or a sitting diastolic blood pressure of 90 mmHg to 100 mmHg prior to administration of MANP.

[0011] In one embodiment, a patient in need of such treatment receives a daily dose of 25 kg / m 2In one embodiment, the patient in need of treatment is a male patient with a waist circumference of 102 cm or greater. In one embodiment, the patient in need of treatment is a female patient with a waist circumference of 88 cm or greater. In one embodiment, the patient in need of treatment has high serum triglyceride levels prior to receiving MANP in accordance with the present disclosure. In one embodiment, the patient in need of treatment has low HDL cholesterol levels prior to receiving MANP in accordance with the present disclosure. In one embodiment, low HDL cholesterol levels are identified in male patients by HDL cholesterol levels of less than 40 mg / dL. In one embodiment, low HDL cholesterol levels are identified in female patients by HDL cholesterol levels of less than 50 mg / dL. In one embodiment, the patient in need of treatment is a patient receiving medication to increase HDL cholesterol levels for low HDL cholesterol levels. In one embodiment, the patient in need of treatment has high fasting plasma glucose levels measured prior to receiving MANP in accordance with the present disclosure. In one embodiment, elevated fasting plasma glucose levels are identified by the patient having a fasting plasma glucose level of 100 mg / dL or greater.

[0012] In one embodiment, the patient in need of such treatment is not pregnant. In one embodiment, the patient in need of such treatment is not breastfeeding. In one embodiment, the patient in need of such treatment does not have a history of alcohol abuse, illicit drug use, severe psychiatric illness, physical dependence on any opioid, or drug abuse or addiction (within two years prior to receiving MANPs according to the methods of the present disclosure). In one embodiment, the patient in need of such treatment does not have a history of difficulty donating blood or have not donated blood or blood products within 45 days prior to administration. In one embodiment, the patient in need of such treatment does not have a history of coronary artery disease, cerebrovascular disease, or syncope. In one embodiment, the patient in need of such treatment does not have a history of epilepsy or other seizure disorder. In one embodiment, the patient in need of such treatment does not have a history of organ transplantation. In one embodiment, the patient in need of such treatment does not have a malignancy within five years prior to receiving MANPs according to the methods of the present disclosure. In one embodiment, the patient in need of such treatment does not have a history of clinically significant intrinsic renal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery.

[0013] The present invention is further disclosed with reference to the accompanying drawings, in which: [Brief explanation of the drawings]

[0014] [Figure 1] The schematic structure of MANP is shown. [Figure 2] In accordance with the present disclosure, a study protocol is outlined to investigate the effects of MANP in hypertensive subjects with metabolic syndrome. [Figure 3A] 1 is a graph plotting circulating levels of ANP-like peptide in patients 24 hours after subcutaneous administration of MANP in accordance with the present disclosure compared to placebo. [Figure 3B] 1 is a graph plotting the change in circulating levels of cGMP over a 24-hour period following subcutaneous administration of MANPs in accordance with the present disclosure compared to placebo. [Figure 4A]1 is a graph plotting the change in sitting systolic blood pressure over 24 hours following subcutaneous administration of MANP in accordance with the present disclosure compared to placebo. [Figure 4B] 1 is a graph plotting the change in sitting diastolic blood pressure over 24 hours following subcutaneous administration of MANP in accordance with the present disclosure compared to placebo. [Figure 4C] 1 is a graph plotting a comparison of the change in heart rate over a 24 hour period following subcutaneous administration of MANPs in accordance with the present disclosure compared to placebo. [Figure 5A] 1 shows the correlation between baseline characteristics and mean systolic blood pressure change in patients undergoing treatment according to the present disclosure. [Figure 5B] 1 shows the correlation between baseline characteristics and mean diastolic blood pressure change in patients undergoing treatment according to the present disclosure. [Figure 6A] 1 is a plot of a comparison of maximum ANP-like peptide elevation after a single MANP injection in male (n=5) and female (n=12) subjects according to the present disclosure. [Figure 6B] 1 is a plot of a comparison of maximum cGMP increase after a single injection of MANP in male (n=5) and female (n=12) subjects according to the present disclosure. [Figure 6C] 1 is a plot of a comparison of the mean systolic blood pressure change after a single injection of MANP in male (n=5) and female (n=12) subjects according to the present disclosure. [Figure 6D] 1 is a plot of a comparison of mean diastolic blood pressure change after a single injection of MANP in male (n=5) and female (n=12) subjects according to the present disclosure. [Figure 7A] 1 shows the correlation between baseline characteristics and maximum ANP-like peptide increase in patients undergoing treatment according to the present disclosure. [Figure 7B] 1 shows the correlation between baseline characteristics and maximum cGMP increase in patients undergoing treatment according to the present disclosure. [Figure 8A]1 is a graph plotting a comparison of the change in HOMA2 insulin sensitivity in patients between baseline and 4 hours after administration of MANPs according to the present disclosure compared to placebo. [Figure 8B] 1 is a graph plotting a comparison of the change in HOMA2 insulin resistance in patients between baseline and 4 hours after administration of MANPs according to the present disclosure compared to placebo. DETAILED DESCRIPTION OF THE INVENTION

[0015] Corresponding reference characters indicate corresponding parts throughout the several views. The examples set forth herein illustrate embodiments of the present disclosure, but should not be construed as limiting the scope of the disclosure in any way.

[0016] This description is not intended to be a detailed catalog of all the different ways in which the present disclosure may be implemented or all the features that may be added to the present disclosure. For example, features shown with respect to one embodiment may be incorporated into other embodiments, and features shown with respect to a particular embodiment may be omitted from that embodiment. Thus, the present disclosure contemplates that, in some aspects of the disclosure, any feature or combination of features described herein may be excluded or omitted. Additionally, numerous modifications and additions to the various aspects suggested herein will become apparent to those skilled in the art in light of the present disclosure without departing from 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. No part of this specification is intended to disclaim any portion of the entire scope of the present disclosure. Accordingly, the following description is intended to illustrate some specific aspects of the present disclosure, but is not intended to exhaustively specify all permutations, combinations, and variations thereof.

[0017] Unless otherwise defined, 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. The terms used to describe the present disclosure herein are used only for the purpose of describing particular aspects or embodiments and are not intended to limit the present disclosure.

[0018] All publications, patent applications, patents and other references cited in this specification are incorporated by reference in their entirety for the teachings related to the sentence and / or paragraph in which the reference is written.The reference to technology in this specification is intended to refer to the technology as generally understood in the technical field, and includes the modification of the technology or the replacement of equivalent technology that is obvious to those skilled in the art.In case of conflict, the present specification, including definitions, shall prevail.

[0019] Unless the context indicates otherwise, it is specifically contemplated that the various features of the present disclosure described herein may be used in any combination. Further, the present disclosure also contemplates that in some aspects of the disclosure, any feature or combination of features described herein may be excluded or omitted.

[0020] The methods disclosed herein comprise and consist of one or more steps or actions for achieving the described method. Method steps and / or actions may be interchanged with one another without departing from the scope of the present disclosure. In other words, unless a specific order of steps or actions is required for proper operation of an embodiment, the order and / or use of specific steps and / or actions may be modified without departing from the scope of the present disclosure. Although methods and materials similar or equivalent to those described herein may be used to implement aspects of the present disclosure, preferred methods and materials are described herein.

[0021] As used in the description of this disclosure and the appended claims, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly dictates otherwise.

[0022] As used herein, "and / or" refers to and includes any and all possible combinations of one or more of the associated listed items, and also includes no combination in the alternative ("or").

[0023] As used herein, the terms "about" and "approximately," when referring to a measurable value such as length, frequency, or duration, are meant to encompass variations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of the specified amount.

[0024] 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, a phrase such as "between about X and Y" means "between about X and about Y," and a phrase such as "about X to Y" means "about X to about Y."

[0025] As used herein, the term "exemplary" is used to mean serving as an example, instance, or illustration. Any aspect or aspects described as "exemplary" should not necessarily be construed as preferred or advantageous over other aspects or aspects, nor is it meant to exclude equivalent structures and techniques known to those skilled in the art. Rather, use of the term "exemplary" is intended to present concepts in a specific form, and the disclosed subject matter is not limited by such examples.

[0026] The present disclosure provides for a method of treating hypertension with metabolic syndrome in a patient, including the method, which may comprise administering a MANP provided herein. In one embodiment, a treatment method according to the present disclosure may beneficially affect or alleviate one or more symptoms associated with the disorder or one or more underlying causes of the disorder. In one embodiment, the present disclosure provides for and includes a method of lowering blood pressure, administered to a patient in need of such treatment, comprising administering a MANP to the patient. In one embodiment, the present disclosure provides for and includes a method of increasing plasma ANP-like peptide levels, administered to a patient in need of such treatment, comprising administering a MANP to the patient. In one embodiment, the present disclosure provides for and includes a method of lowering plasma glucose levels, administered to a patient in need of such treatment, comprising administering a MANP to the patient. In one embodiment, the present disclosure provides for and includes a method of improving insulin sensitivity, administered to a patient in need of such treatment, comprising administering a MANP to the patient. In one embodiment, the present disclosure provides for and includes a method of increasing plasma NEFA levels, the method being administered to a patient in need of such treatment, comprising administering MANP to the patient. In one embodiment, the present disclosure provides for and includes a method of treating a patient with a low baseline plasma cGMP level. In one embodiment, a low baseline plasma cGMP level is a plasma cGMP level below 10 pmol / mL (e.g., below 5 pmol / mL, below 1 pmol / mL, below 0.5 pmol / mL, below 0.1 pmol / mL, below 0.05 pmol / mL, or below 0.01 pmol / mL).

[0027] As used herein, the term "treat" or "treatment" refers to an approach to obtaining a beneficial or desired clinical result. In one embodiment, the term "treat" or "treatment" refers to administering a MANP disclosed herein to partially or completely alleviate, ameliorate, reduce, inhibit, delay the onset, reduce the severity, and / or reduce the incidence of one or more symptoms, characteristics, or causes of hypertension with metabolic syndrome. The term "treat" or "treatment" includes administering a MANP disclosed herein to prevent or delay the onset of symptoms, complications, or biochemical indicators of hypertension with metabolic syndrome, alleviate symptoms, or prevent or inhibit further development of hypertension with metabolic syndrome. Treatment can be prophylactic (preventing or delaying the onset of hypertension with metabolic syndrome) or therapeutically suppressing or alleviating symptoms after the onset of hypertension with metabolic syndrome.

[0028] In one embodiment, the present disclosure includes methods of treatment that may 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 disorder. In one embodiment, the present disclosure includes methods of treatment that may 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 disorder. In one embodiment, methods of treatment according to the present disclosure may 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 disorder. In one embodiment, the present disclosure includes methods of treatment that may 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 disorder.

[0029] As used herein, "hypertension" refers to a systolic blood pressure of 140 millimeters of mercury (mmHg) to 180 mmHg or a diastolic blood pressure of 90 mmHg to 100 mmHg in a sitting position. In one embodiment, hypertension is a blood pressure measurement of 130 mmHg systolic blood pressure, 80 mmHg diastolic blood pressure, or higher.

[0030] In one embodiment, blood pressure is measured by a sphygmomanometer (blood pressure monitor) or sphygmomanometer. In one embodiment, the sphygmomanometer (blood pressure monitor) includes an inflatable cuff, a measuring portion (i.e., a mercury manometer or an aneroid gauge), and a pressurizing mechanism, which may be a manually operated rubber bulb and valve or an electrically operated pump. In one embodiment, the sphygmomanometer includes a manual meter. In one embodiment, the sphygmomanometer includes a digital meter.

[0031] In one embodiment, sitting blood pressure is measured while the subject is seated in a comfortable chair with a backrest for at least 5 minutes prior to measurement. The subject's feet are flat on the ground, not crossed. The subject's arm, with the cuff attached, is placed on a table at chest height.

[0032] In one embodiment, systolic and diastolic blood pressures are measured using methods known in the art. In one embodiment, while the subject is seated with their arm supported, a sphygmomanometer cuff is smoothly and snugly placed around the upper arm at approximately the same vertical height as the heart. In one embodiment, the cuff size is selected and fitted to the patient. In one embodiment, measurements are taken from both arms to determine whether the pressure is significantly higher in one arm than the other. A difference of 10 mmHg may be an indication of aortic stenosis. If the arm measurements differ, the arm with the higher reading will be used for subsequent measurements. The cuff is inflated until the artery is completely occluded. In one embodiment, while listening to the brachial artery with a stethoscope, the examiner uses a manual instrument to slowly release the cuff pressure at a rate of approximately 2 mmHg per heartbeat. As the pressure in the cuff drops, a "whoosh" or thumping sound is heard as blood flow resumes in the artery. The pressure at which this sound begins is noted and recorded as the systolic blood pressure. The cuff pressure is further released until the sound is no longer audible. This is recorded as diastolic blood pressure. In one embodiment, digital instruments use a cuff that may be worn around the upper arm, wrist, or finger, depending on the instrument, and in each case is level with the heart. Digital instruments measure blood pressure oscillometrically by inflating the cuff and gradually releasing the pressure, just like manual instruments.

[0033] In one embodiment, the presence or degree of hypertension may be assessed using methods known in the art, such as routine clinical examinations to assess blood pressure, 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 diastolic filling pressure, and diastolic potentiation by pulse wave and tissue Doppler; measurement of cardiac output, pulmonary wedge capillary pressure, pulmonary artery pressure, right ventricular pressure, right atrial pressure, and systemic and pulmonary vascular resistance using a Swan-Ganz catheter; assessment of renal function by measuring glomerular filtration rate, serum creatinine, and blood urea nitrogen; and type B naturopathies. These include, but are not limited to, measurement of biomarkers such as brain natriuretic peptide (BNP), amino-terminal pro-BNP (NT-proBNP), troponin T, troponin I, C-reactive protein (CRP), creatine kinase, serum cystatin C, albuminuria, neutrophil gelatinase-associated lipocalin (NGAL), N-acetyl-β-D-glucosaminidase (NAG), kidney injury molecule-1 (KIM-1), angiotensin II, renin, aldosterone, and inflammatory cytokines (e.g., interleukin (IL)-6, IL-18).

[0034] As used herein, "metabolic syndrome" refers to a condition diagnosed based on the criteria established by the National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ATP III). Specifically, the NCEP ATP III defines metabolic syndrome as the presence of any two of the following characteristics: 1. Abdominal obesity: Waist circumference of 102 cm (40 inches) or more for men and 88 cm (35 inches) or more for women; 2. Serum triglyceride level is 150 mg / dL (1.7 mmol / L) or higher, 3. Serum high-density lipoprotein (HDL) cholesterol level is less than 40 mg / dL (1 mmol / L) for men and less than 50 mg / dL (1.3 mmol / L) for women, or you are currently receiving medical treatment for low HDL cholesterol levels. 4. Fasting plasma glucose level ≥ 100 mg / dL (5.6 mmol / L) or medication for elevated blood glucose levels. In one embodiment, metabolic syndrome is identified in patients with three or more of the risk factors defined by the American Heart Association, including high blood sugar, low HDL cholesterol, high triglyceride levels, large waist circumference, and high blood pressure. In one embodiment, a lipid profile or lipid panel is a panel of blood tests used to detect abnormalities in lipids such as cholesterol and triglycerides. Typically, laboratories measure only three values: total cholesterol, HDL, and triglycerides. LDL may be calculated from these three data points. In one embodiment, laboratories require patients to fast for 9 to 12 hours before screening. In one embodiment, some diagnostic laboratories accept non-fasting samples.

[0035] In one aspect, the method of treatment provided herein is a method of achieving lower blood pressure, enhanced insulin sensitivity, elevated plasma nonesterified fatty acids (NEFA), lowered plasma glucose levels, increased natriuresis, arterial vasodilation, suppression of renin and aldosterone, reduced apoptosis and hypertrophy, increased lucitropy, induction of vascular regeneration, increased lipolysis, and browning of white adipocytes in a patient in need of such treatment.

[0036] MANP MANP, an ANP analog, is a guanylyl 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 peptide with a unique 12-amino acid C-terminal extension, including the 28 amino acid residues of native ANP. Compared to ANP, MANP is more resistant to degradation, has enhanced binding to the GC-A receptor, causes increased and more sustained sodium excretion, and causes more sustained suppression of aldosterone.

[0037] The methods provided herein may, in part, involve treating a patient with MANP. The present disclosure provides polypeptides or nucleic acids encoding the MANP polypeptides listed in Table 1. As used herein, "MANP" may have the amino acid sequence set forth in SEQ ID NO:3 or may be a variant of the sequence set forth in SEQ ID NO:3. As depicted in FIG. 1, MANP is an ANP-based peptide having an amino acid sequence that includes the 28-amino acid mature human ANP sequence (SLRRSSCFGGRMDRIGAQSGLGCNSFRY, SEQ ID NO:1) plus an additional 12 amino acids at the carboxy terminus (RITAREDKQGWA, SEQ ID NO:2). The full-length sequence of MANP is SLRRSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA (SEQ ID NO:3). A representative nucleic acid sequence encoding a MANP is 5'-agcctgcggagatccagctgcttcgggggcaggatggacaggattggagcccagagcggactgggctgtaacagcttccggtaccggataacagccagggaggacaagcagggctgggcctag-3' (SEQ ID NO: 4). [Table 1]

[0038] In one embodiment, a MANP used in the methods provided herein may comprise the entire amino acid sequence set forth in SEQ ID NO: 3. In one embodiment, a MANP comprises the amino acid sequence set forth in SEQ ID NO: 3, which may comprise 1, or in the range of 1 to 10 (e.g., 10, 1 to 9, 2 to 9, 1 to 8, 2 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 2, or 1) amino acid additions, deletions, and / or substitutions. In one embodiment, a MANP comprises the amino acid sequence set forth in SEQ ID NO: 3 and may further have a single amino acid residue addition, deletion, or substitution of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 residues. In one embodiment, a MANP may have one or more additions, deletions, 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, in which threonine is deleted (SLRRSSCFGGRMDRIGAQSGLGCNSFRYRIAREDKQGWA, SEQ ID NO:5), tryptophan is substituted with tyrosine (SLRRSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGYA, SEQ ID NO:6), serine is added between the lysine and glutamine (SLRRSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKSQGWA, SEQ ID NO:7), or any combination thereof. In one embodiment, the MANP may 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 one embodiment, the MANP may have one or more additions, deletions, 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 a polypeptide having the amino acid sequence set forth in SEQ ID NO: 3, in which four amino acids derived from urodilatin have been added to the N-terminus (TAPRSLRRSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA, SEQ ID NO: 9), a polypeptide in which the arginine residues at positions 3 and 4 have been replaced with lysine residues (SLKKSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA, SEQ ID NO: 10), a polypeptide in which the D-isoform of serine has been replaced at position 6 (SLRRSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA, SEQ ID NO: 11), and a polypeptide in which the D-isoform of serine has been replaced at position 6 (SLRRSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA, SEQ ID NO: 12). 1), a substitution of the D-isoform of arginine at position 4 and deletion of serine at position 5 (SLRRSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA, SEQ ID NO: 12), a substitution of serine residues at positions 1, 5, and 6 with threonine residues (TLRRTTCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA, SEQ ID NO: 13), a substitution of leucine at position 2 with tryptophan (SWRRSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQGWA, SEQ ID NO: 14), or any combination thereof.

[0039] In one embodiment, any amino acid residue set forth in SEQ ID NO: 3 may be deleted, or 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) may be added to the sequence set forth in SEQ ID NO: 3. In one embodiment, a MANP may include one or more chemical structures, such as ε-aminohexanoic acid, hydroxylated amino acids (such as 3-hydroxyproline, 4-hydroxyproline, (5R)-5-hydroxy-L-lysine, allo-hydroxylysine, 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.

[0040] In one embodiment, MANPs having one or more amino acid additions, deletions, or substitutions relative to the representative MANP sequence set forth in SEQ ID NO: 3 (also referred to herein as "mutant" MANPs) may be generated using any suitable method. In one embodiment, amino acid substitutions may be made by selecting substitutions that do not significantly alter their effect on maintaining (a) the structure of the peptide backbone in the area of ​​the substitution, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain. In one embodiment, naturally occurring residues are divided into groups based on side chain properties. (1) hydrophobic amino acids (norleucine, methionine, alanine, valine, leucine, and isoleucine), (2) neutral hydrophilic amino acids (cysteine, serine, and threonine), (3) acidic amino acids (aspartic acid and glutamic acid), (4) basic amino acids (asparagine, glutamine, histidine, lysine, and arginine), (5) amino acids that affect chain orientation (glycine and proline), and (6) aromatic amino acids (tryptophan, tyrosine, and phenylalanine). Substitutions made within these groups may be considered conservative substitutions. Non-limiting examples of useful conservative substitutions may include, but are not limited to, substitutions of valine for alanine, lysine for arginine, glutamine for asparagine, glutamic acid for aspartic acid, serine for cysteine, asparagine for glutamine, aspartic acid for glutamic acid, proline for glycine, arginine for histidine, leucine for isoleucine, isoleucine for leucine, arginine for lysine, leucine for methionine, leucine for phenylalanine, glycine for proline, threonine for serine, serine for threonine, tyrosine for tryptophan, phenylalanine for tyrosine, and / or leucine for valine.

[0041] 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. [Table 2]

[0042] In one aspect, MANP can contain one or more non-conservative substitutions. Non-conservative substitutions usually involve replacing a member of one of the above classes with a member of another class. Such production may be desirable to provide such compounds in large quantities or to provide alternative embodiments of such compounds. Whether an amino acid change results in a functional polypeptide can be easily determined by analyzing the specific activity of the polypeptide variant, for example, using cGMP radioimmunoassay (RIA) or cGMP ELISA assay.

[0043] In one embodiment, 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 one embodiment, a MANP can include the amino acid sequence set forth in SEQ ID NO: 3, but with a specific number of amino acid substitutions. In one embodiment, 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, MANPs in which one or more L-amino acids in the N-terminal region of the polypeptide are substituted with D-amino acids (e.g., a D-serine residue at position 6 as set forth in SEQ ID NO: 11, or a D-arginine at position 4 as set forth in SEQ ID NO: 12).

[0044] In one embodiment, a MANP may comprise 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 percentage of sequence identity is calculated by aligning the amino acid sequences to determine the number of matching positions, dividing the number of matching positions by the total number of aligned amino acids, and multiplying the result by 100. A matching position refers to a position where an identical amino acid occurs at the same position in the aligned amino acid sequences. The percentage of sequence identity may also be determined for any nucleic acid sequence.

[0045] The percentage of 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 indicated by a particular sequence identification number using the BLAST2 sequence (Bl2seq) program from the standalone version of BLASTZ, which includes BLASTN version 2.0.14 and BLASTP version 2.0.14. This standalone version of BLASTZ can be obtained online at fr.com / blast or ncbi.nlm.nih.gov. Instructions for using the Bl2seq program can be found in the readme file provided with BLASTZ. Bl2seq performs a comparison between two sequences using either the BLASTN or BLASTP algorithm. BLASTN is used to compare nucleic acid sequences, and BLASTP is used to compare amino acid sequences. To compare two nucleic acid sequences, set the options as follows: -i is set to a file containing the first nucleic acid sequence to be compared (e.g., C:\seq1.txt), -j is set to a 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 embodiment, the following command may be used to generate an output file containing the comparison between the two sequences: C:\Bl2seq -ic:\seq1.txt -jc:\seq2.txt -p blastn -oc:\output.txt -q -1 -r 2. To compare two amino acid sequences, the Bl2seq options 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 embodiment, the following command may be used to generate an output file containing a comparison between two amino acid sequences: C:\Bl2seq -ic:\seq1.txt -jc:\seq2.txt -p blastp -oc:\output.txt. If the two compared sequences share homology, the specified output file will present the homologous regions as aligned sequences. If the two compared sequences do not share homology, the specified output file will not present aligned sequences.

[0046] The number of matches is determined by counting the number of positions where the same nucleotide or amino acid residue is present in both sequences when aligned. The percentage of sequence identity is determined by dividing the number of matches by the length of the sequence shown in the identified sequence (e.g., SEQ ID NO: 3) or by the connected length (e.g., 20 consecutive nucleotides or amino acid residues from the sequence shown in the identified sequence), and then multiplying the resulting value by 100. In one embodiment, an amino acid sequence that has 37 matches when aligned with the sequence set forth in SEQ ID NO: 3 is 92.5 percent identical to the sequence set forth in SEQ ID NO: 3 (i.e., 37÷40×100=92.5). Please note that the percentage of sequence identity value is rounded to one decimal place. In one embodiment, 75.11, 75.12, 75.13, and 75.14 are rounded down to 75.1, and 75.15, 75.16, 75.17, 75.18, and 75.19 are rounded up to 75.2. Note also that length values ​​are always integers.

[0047] Isolated MANPs may be manufactured using any suitable method, including solid-phase synthesis, and may be produced using manual or automated techniques (e.g., using an Applied BioSystems (Foster City, Calif.) peptide synthesizer or a Biosearch Inc. (San Rafael, Calif.) automated peptide synthesizer). Disulfide bonds between cysteine ​​residues may also be introduced by mild oxidation of linear polypeptides with KCN, as described, for example, in U.S. Pat. No. 4,757,048. MANPs may also be produced recombinantly.

[0048] The MANPs described herein are typically cyclic via disulfide bonds between cysteine ​​residues (see Figure 1). In one embodiment, the sulfhydryl group on the cysteine ​​residue may be replaced with an alternative group (e.g., -CHCH-). To replace the sulfhydryl group with a -CH- group, in one embodiment, the cysteine ​​residue may be substituted with alpha-aminobutyric acid. Such cyclic analog polypeptides may, in one embodiment, be produced according to the method of Lebl and Hruby ((1984) Tetrahedron Lett., 25:2067-2068) or using the procedures disclosed in U.S. Pat. No. 4,161,521.

[0049] In one embodiment, an ester bridge may be formed by reacting the OH of serine or threonine with the carboxyl group of aspartic acid or glutamic acid, resulting in a bridge having the structure -CHCOCH-. Similarly, an amide may be obtained by reacting the side chain of lysine with aspartic acid or glutamic acid, resulting in a bridge having the structure -CHC(O)NH(CH)-. Methods for synthesizing these bridges are known in the art (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 embodiment, one method for preparing an ester of a polypeptide of the invention, when using the Merrifield synthesis technique, is to cleave the completed polypeptide from the resin in the presence of the desired alcohol, which is performed under basic or acidic conditions depending on the resin. The C-terminus of a polypeptide may be esterified directly upon release from the resin without isolating the free acid. Amides of polypeptides may also be prepared using techniques for converting carboxylic acid groups or precursors to amides (e.g., techniques known in the art). One method for forming amides at the C-terminal carboxyl group includes cleaving the polypeptide from the 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 bridge-forming amino acid residues and reactions are described in U.S. Pat. No. 4,935,492. The preparation of peptide analogs containing nonpeptidyl bonds linking amino acid residues is also known in the art.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. al. (1982), Tetrahedron Lett., 23:2533, EP45665, Holladay et al. (1983), Tetrahedron Lett., 24:4401, and Hruby (1982), Life Sci., 31:189.

[0050] N-acyl derivatives of amino groups in polypeptides can be prepared by using an N-acyl-protected amino acid in the final condensation or by acylating a protected or unprotected peptide. O-acyl derivatives can be prepared, in one embodiment, by acylating a free hydroxypeptide or peptide resin. Either acylation can be carried out using standard acylating reagents such as acyl halides, anhydrides, or acylimidazoles. If desired, both N- and O-acylation can be carried out simultaneously.

[0051] In one embodiment, the MANP may be pegylated, acetylated, or both. In one embodiment, the polypeptide may be covalently attached to an oligomer, such as a short, amphiphilic oligomer, which allows administration or improves the pharmacokinetic or pharmacodynamic profile of the attached polypeptide. The oligomer may comprise water-soluble polyethylene glycol (PEG) and / or a lipid-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 may be attached to the free amino terminus or any lysine side chain (epsilon amino group), and the lysine residue for this attachment may be located at either the C-terminus or N-terminus of the peptide. Conjugation to PEG or other suitable polymers, or fusion with albumin or other suitable polypeptides, may result in a modified MANP with an extended half-life compared to the unmodified MANP. Without being bound by a particular mechanism, the increased serum half-life of the modified MANP may be due to reduced protease degradation, reduced immune recognition, or reduced cellular scavenging. Methods for modifying polypeptides by linking to PEG (also known as "PEGylation") or other polymers are known in the art and are 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. Each of these documents is incorporated herein by reference in its entirety.Methods for modifying polypeptides by fusion with albumin are also known in the art, including U.S. Patent Publication No. 2004 / 0086976 and Wang et al. (2004), Pharm. Res., 21:2105-2111, each of which is incorporated herein by reference in its entirety.

[0052] In one embodiment, the MANP may be fused to the Fc domain of an immunoglobulin molecule (e.g., an IgG1 molecule), in which case active transport of the fusion polypeptide occurs across an epithelial cell barrier via the Fc receptor. In one embodiment, the polypeptide may be a cyclic polypeptide. A cyclic polypeptide may be obtained by linking cysteine ​​residues. However, in one embodiment, it is also contemplated to replace the sulfhydryl group on the cysteine ​​residue with another group, such as -CH2-CH2-. In one embodiment, the cysteine ​​residue may be replaced with the analogous alpha-aminobutyric acid to replace the sulfhydryl group with a -CH2- group. These cyclic analog peptides may be formed, in one embodiment, according to the method of Lebl and Hruby, supra, or by employing the procedure disclosed in U.S. Pat. No. 4,161,521.

[0053] Salts of carboxyl groups of MANPs may, in one embodiment, be prepared by contacting the polypeptide with one or more equivalents of a desired base, such as 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, etc.). Acid addition salts of the polypeptide may be prepared by contacting the polypeptide with one or more equivalents of an inorganic or organic acid (e.g., hydrochloric acid).

[0054] As used herein, the term "polypeptide" refers to a compound of two or more subunit amino acids, regardless of post-translational modification (e.g., phosphorylation or glycosylation). The subunits may be linked by peptide bonds or, in one embodiment, other bonds such as ester or ether bonds. The term "amino acid" refers to any natural and / or unnatural or synthetic amino acid, including D / L optical isomers.

[0055] The term "isolated," as used herein with respect to a polypeptide, means that the polypeptide (1) is not associated with naturally occurring proteins, (2) is free of other proteins from the same source (e.g., free of human proteins), (3) is expressed by cells of a different species, or (4) is not found in nature. An isolated polypeptide may, in one embodiment, be encoded by DNA or RNA, including synthetic DNA or RNA, or a combination thereof.

[0056] The term "substantially pure" as used herein in reference to a polypeptide means that the polypeptide is substantially free from other polypeptides, lipids, carbohydrates, and nucleic acids with which it is naturally associated. A substantially pure polypeptide may be any polypeptide that has been removed from its natural environment and is at least 60% pure. A substantially pure polypeptide is at least about 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% pure, or about 65-75%, 75-80%, 80-85%, 85-90%, 90-95%, or 95-99% pure. Typically, a substantially pure polypeptide will yield a single major band on a non-reducing polyacrylamide gel. In one embodiment, a substantially pure polypeptide may be a chemically synthesized polypeptide.

[0057] Any method may be used to obtain a substantially pure polypeptide. In one embodiment, polypeptide purification techniques such as affinity chromatography and high-performance liquid chromatography (HPLC), as well as general polypeptide synthesis techniques, may be used. Any material may be used as a source for obtaining a substantially pure polypeptide. In one embodiment, tissue from wild-type or transgenic animals may be used as the starting material. Alternatively, tissue culture cells engineered to overexpress a specific polypeptide may be used to obtain a substantially pure polypeptide. Furthermore, the polypeptide may be engineered to contain an amino acid sequence that allows it to be captured on an affinity matrix. In one embodiment, a tag, such as c-myc, hemagglutinin, polyhistidine, or Flag™ tag (Kodak), may be used to aid in polypeptide purification. Such tags may be inserted anywhere within the polypeptide, including at either the carboxyl or amino terminus, or anywhere in between. Other fusions that may be used include enzymes, such as alkaline phosphatase, that aid in polypeptide detection.

[0058] MANPs (e.g., mutant MANPs having conservative and / or non-conservative substitutions with respect to SEQ ID NO: 3), as well as fragments of SEQ ID NO: 3 or mutant MANPs (e.g., fragments of any of SEQ ID NOs: 3-14), can be screened for biological activity using any of a number of assays. In one embodiment, the activity of a MANP may be determined in vitro by testing its effect on cGMP production in cultured cells (e.g., cultured cardiac fibroblasts, aortic endothelial cells, or glomerular cells). Cells are incubated with MANPs (e.g., 10 -10 ~10 -4 The samples may be exposed to M MANP and the samples may be assayed to assess the effect of the polypeptide on cGMP production. cGMP production may, in one embodiment, be detected and measured using a competitive RIA cGMP kit (Perkin-Elmer, Boston, MA).

[0059] In one embodiment, the activity of MANPs may be assessed in vivo after administration to a patient (e.g., a human, non-human primate, rodent, dog, cat, pig, sheep, horse, or cow) by examining its effect on factors such as plasma cGMP levels, urinary cGMP excretion, net renal cGMP production, glomerular filtration rate, hemodynamic functions such as blood pressure, heart rate, cardiac output, pulmonary artery wedge pressure, systemic vascular resistance, and renal functions such as renal blood flow, urine volume, and sodium excretion rate. In one embodiment, such parameters may be assessed after inducing hypertension in a patient.

[0060] As used herein, the term "nucleic acid" encompasses both RNA and DNA, including cDNA, genomic DNA, and synthetic (e.g., chemically synthesized) DNA. Nucleic acids may be double-stranded or single-stranded. If the nucleic acid is single-stranded, it may be the sense strand or the antisense strand. Furthermore, nucleic acids may be circular or linear.

[0061] The term "isolated," as used herein with respect to nucleic acids, refers to a naturally occurring nucleic acid that is not immediately adjacent to both sequences (5' and 3') that it is immediately adjacent to in the naturally occurring genome of the organism from which it originates. In one embodiment, an isolated nucleic acid can be, but is not limited to, a recombinant DNA molecule of any length, so long as one of the nucleic acid sequences normally found immediately adjacent to the recombinant DNA molecule in the naturally occurring genome has been removed or is absent. Thus, isolated nucleic acids include, but are not limited to, recombinant DNA that exists as a separate molecule independent of other sequences (e.g., cDNA or genomic DNA fragments produced by PCR or restriction endonuclease treatment), as well as recombinant DNA that is integrated into a vector, autonomously replicating plasmid, virus (e.g., retrovirus, adenovirus, or herpesvirus), or genomic DNA of a prokaryote or eukaryote. Furthermore, isolated nucleic acids can include recombinant DNA molecules that are part of hybrid or fusion nucleic acid sequences.

[0062] The term "isolated" as used herein with respect to nucleic acids also encompasses non-naturally occurring nucleic acids, because non-naturally occurring nucleic acid sequences are not found in nature and do not have immediately adjacent sequences in a naturally occurring genome. In one embodiment, non-naturally occurring nucleic acids, such as artificial nucleic acids, are considered isolated nucleic acids. Artificial nucleic acids may be made using common molecular cloning or chemical nucleic acid synthesis techniques. Isolated non-naturally occurring nucleic acids may be independent of other sequences or may be integrated into a vector, an autonomously replicating plasmid, a virus (e.g., retrovirus, adenovirus, herpesvirus), or the genomic DNA of a prokaryote or eukaryote. Furthermore, non-naturally occurring nucleic acids may include nucleic acid molecules that are part of a hybrid or fusion nucleic acid sequence.

[0063] It will be apparent to one of skill in the art that, in one embodiment, a nucleic acid present among hundreds to millions of other nucleic acid molecules in a cDNA library, a genomic library, or a gel slice containing a genomic DNA restriction digest is not considered an isolated nucleic acid.

[0064] In one embodiment, the 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 hybridization conditions to the 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 hybridization conditions can be moderately or highly stringent.

[0065] As used herein, "moderately stringent hybridization conditions" refer to a hybridization condition that is performed in a medium containing 25 mM KPO (pH 7.4), 5X SSC, 5X Denhardt's solution, 50 μg / mL denatured sonicated salmon sperm DNA, 50% formamide, 10% dextran sulfate, and 1-15 ng / mL probe (approximately 5 × 10 7 This means that hybridization is carried out at about 42°C in a hybridization solution containing 2× SSC and 0.1% sodium dodecyl sulfate, and washing is carried out at about 50°C in a wash solution containing 2× SSC and 0.1% sodium dodecyl sulfate.

[0066] "Highly stringent hybridization conditions" refer to 25 mM KPO (pH 7.4), 5X SSC, 5X Denhardt's solution, 50 μg / mL denatured sonicated salmon sperm DNA, 50% formamide, 10% dextran sulfate, and 1–15 ng / mL probe (approximately 5 × 10 7This means that hybridization is carried out at about 42°C in a hybridization solution containing 0.2X SSC and 0.1% sodium dodecyl sulfate, and washing is carried out at about 65°C in a washing solution containing 0.2X SSC and 0.1% sodium dodecyl sulfate.

[0067] 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 one embodiment, polymerase chain reaction (PCR) techniques can be used to obtain isolated nucleic acids containing nucleotide sequences encoding the MANPs provided herein. PCR is a procedure or technique for enzymatically amplifying target nucleic acids. Typically, sequence information from the ends of the region of interest or beyond is used to design oligonucleotide primers identical in sequence to opposite strands of the template to be amplified. PCR can also be used to amplify specific sequences from DNA and RNA, including sequences derived from total genomic DNA or total cellular RNA. Primers are typically 14 to 40 nucleotides in length but can range from 10 to several hundred nucleotides in length. General PCR techniques are described, in one embodiment, in "PCR Primer: A Laboratory Manual," ed. by Dieffenbach and Dveksler, Cold Spring Harbor Laboratory Press, 1995. When RNA is used as the template, a complementary DNA (cDNA) strand can be synthesized using reverse transcriptase. To obtain isolated nucleic acid, ligase chain reaction, strand displacement amplification, self-sustained sequence replication or nucleic acid sequence-based amplification may be used.In one embodiment, see 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.

[0068] Isolated nucleic acids encoding MANPs can also be chemically synthesized as single nucleic acid molecules (e.g., using automated DNA synthesis in the 3' to 5' direction using phosphoramidite technology) or as a series of oligonucleotides. In one embodiment, one or more long oligonucleotide pairs (e.g., more than 100 nucleotides) containing the desired sequence can be synthesized, with each pair containing a short segment of complementarity (e.g., about 15 nucleotides) such that a double-stranded sequence is formed when the oligonucleotide pair is annealed. DNA polymerase can be used to extend the oligonucleotides, resulting in one double-stranded nucleic acid molecule per oligonucleotide pair, which can then be ligated into a vector.

[0069] Additionally, isolated nucleic acids encoding MANPs may be obtained by mutagenesis. In one embodiment, a reference sequence may be mutated using standard techniques, including oligonucleotide-directed mutagenesis and PCR-mediated site-specific mutagenesis. See Short Protocols in Molecular Biology, Chapter 8, Green Publishing Associates and John Wiley & Sons, edited by Ausubel et al., 1992. Non-limiting examples of mutant MANPs are provided herein.

[0070] Also provided are vectors containing nucleic acids such as those described herein.A "vector" is a replicon, such as a plasmid, phage, or cosmid, into which another DNA segment can be inserted so as to cause replication of the inserted segment.An "expression vector" is a vector that contains 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.

[0071] In an expression vector, a nucleic acid (e.g., a nucleic acid encoding a MANP) may be operably linked to one or more expression control sequences. As used herein, "operably linked" means that the expression control sequence is incorporated into a genetic construct so that it effectively controls the expression of a coding sequence of interest. Examples of expression control sequences include promoters, enhancers, and transcription termination regions. A promoter is an expression control sequence consisting of a region of a DNA molecule and is usually located within 100 to 500 nucleotides upstream of the transcription start site (usually near the RNA polymerase II initiation site). To place a coding sequence under the control of a promoter, the translation start site of the translation reading frame of the polypeptide must be located 1 to approximately 50 nucleotides downstream of the promoter. Enhancers provide expression specificity in terms of time, location, and level. Unlike promoters, enhancers can function even when located at various distances from the transcription site. Enhancers may also be located downstream of the transcription start site. A coding sequence is "operably linked" and "under the control" of an expression control sequence in a cell if RNA polymerase is capable of transcribing the coding sequence into mRNA, which can be translated into the protein encoded by the coding sequence. Thus, expression vectors can be useful for producing not only antibodies, but also other multivalent molecules.

[0072] Suitable expression vectors include, but are not limited to, plasmids and viral vectors, which in one embodiment are derived from bacteriophage, baculovirus, tobacco mosaic virus, herpes virus, cytomegalovirus, retrovirus, vaccinia virus, adenovirus and adeno-associated virus. Numerous vectors and expression vectors are commercially available from companies such as Novagen (Madison, Wisconsin, USA), Clontech (Palo Alto, California, USA), Stratagene (La Jolla, California, USA), and Invitrogen / Life Technologies (Carlsbad, California, USA).

[0073] Expression vectors may contain 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™ tag (Kodak, New Haven, Connecticut, USA) sequences, are typically expressed as fusions with the encoded polypeptide. Such tags may be inserted anywhere within the polypeptide, including at the carboxyl or amino terminus.

[0074] Host cells containing the vectors are also provided. The term "host cell" is intended to include prokaryotic and eukaryotic cells into which a recombinant expression vector (e.g., a vector encoding MANP) can be introduced. As used herein, "transformation" and "transfect" encompass the introduction of a nucleic acid molecule (e.g., a vector) into a cell by one of many techniques. Without being limited to a particular technique, many of these techniques are established in the art. In one embodiment, suitable methods for transforming and transfecting host cells are described in Sambrook et al., Molecular Cloning: A Laboratory Manual (2002). nd(edition), Cold Spring Harbor Laboratory, New York (1989). In one embodiment, calcium phosphate precipitation, electroporation, heat shock, lipofection, microinjection, and virus-mediated nucleic acid transfer may be used to introduce nucleic acids into cells. Furthermore, naked DNA may be delivered directly to cells in vivo, as described elsewhere (U.S. Patent Nos. 5,580,859 and 5,589,466, the entire contents of which are incorporated herein by reference). Note that although host cells may express the encoded polypeptide, cells containing the isolated nucleic acid molecules provided herein need not express the polypeptide. The isolated nucleic acid molecules transformed into host cells may be integrated into the cell's genome or maintained in an episomal state. Thus, host cells may be stably or transiently transfected with constructs containing the isolated nucleic acid molecules provided herein.

[0075] The isolated nucleic acid molecule can be introduced into cells in vivo or in vitro using suitable methods.In one embodiment, calcium phosphate precipitation, electroporation, heat shock, lipofection, microinjection, and virus-mediated nucleic acid transfer are methods that can be used to introduce the isolated nucleic acid molecule into cells.In addition, naked DNA can be directly delivered to cells in vivo, as described elsewhere (for example, U.S. Patent Nos. 5,580,859 and 5,589,466, and their continuations).In addition, the isolated nucleic acid molecule can be introduced into cells by creating transgenic animals.

[0076] Any suitable method may be used to identify cells containing an isolated nucleic acid molecule encoding a MANP, including, but not limited to, PCR and nucleic acid hybridization techniques such as Northern and Southern analysis. In one embodiment, immunohistochemistry and biochemistry techniques may be used to determine whether a cell contains a particular isolated nucleic acid molecule by detecting the expression of the polypeptide encoded by the nucleic acid molecule.

[0077] The isolated MANPs and nucleic acids encoding MANPs described herein can be used to treat patients identified as having conditions such as hypertension with metabolic syndrome. Accordingly, one or more MANPs—or nucleic acids encoding one or more MANPs—can be incorporated into compositions for administration to patients (e.g., patients with or at risk of having hypertension, resistant hypertension (RH), and / or cardiorenal disease). Any suitable method may be used to formulate and subsequently administer the compositions. Dosages typically depend on the patient's responsiveness to the administered agent, with treatment duration lasting from several days to several months or longer, or until a suitable response is achieved. Those skilled in the art routinely determine optimal dosages, administration methods, and repetition rates. Optimal dosages may vary depending on the relative potency of each agent (e.g., MANPs and Fs) and generally will be determined based on the EC2A1 / ... 50 The estimated dose may be based on the following: Compositions comprising one or more MANPs described herein may be administered one or more times daily, weekly, monthly, or even less frequently, or may be administered continuously for a period of time (e.g., hours, days, or weeks). In one embodiment, MANPs, or compositions comprising MANPs, may be administered to a patient at a dose of at least about 0.01 ng of MANP per kg of body weight to about 100 mg of MANP per kg of body weight. In one embodiment, MANPs, or compositions comprising MANPs, may be administered as an infusion for 1 to 30 days or more (e.g., at a dose of about 1 pmol MANP / kg / min to about 500 nmol MANP / kg / min).

[0078] One or more MANPs, or nucleic acids encoding one or more MANPs, may be mixed, encapsulated, conjugated, or otherwise associated with each other and / or with other molecules, molecular structures, or compound mixtures (e.g., in one embodiment, liposomes, receptor or cell targeting molecules, or oral, topical, or other formulations to aid uptake, distribution, and / or absorption, etc.).

[0079] In one embodiment, the compositions of the present disclosure may include a MANP (or a nucleic acid encoding a MANP) in combination with a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers, in one embodiment, include pharmaceutically acceptable solvents, suspending agents, or any other pharmacologically inert vehicles for delivering polypeptides and / or other compounds to a subject. Pharmaceutically acceptable carriers may be liquid or solid and may be selected to provide the desired bulk, appearance, and other relevant transport and chemical properties when combined with one or more therapeutic compounds and any other components of a given pharmaceutical composition, taking into account the intended method of administration. Useful pharmaceutically acceptable carriers include, but are not limited to, water, saline, binders (e.g., polyvinylpyrrolidone or hydroxypropylmethylcellulose), fillers (e.g., lactose or dextrose and other sugars, gelatin, or calcium sulfate), lubricants (e.g., starch, polyethylene glycol, or sodium acetate), disintegrants (e.g., starch or sodium starch glycolate), and wetting agents (e.g., sodium lauryl sulfate).

[0080] Compositions and formulations for parenteral, intrathecal, or intraventricular administration include sterile aqueous solutions (e.g., sterile physiological saline), which may also contain buffers, diluents, and other suitable additives (e.g., penetration enhancers, carrier compounds, and other pharmaceutically acceptable carriers). Compositions and formulations for oral administration, in one embodiment, include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets, or tablets. Such compositions may also incorporate thickeners, flavoring agents, diluents, emulsifiers, dispersing aids, or binders.

[0081] Pharmaceutical compositions include, but are not limited to, solutions, emulsions, aqueous suspensions, and liposome-containing formulations. In one embodiment, these compositions can be produced from various components, including 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 high solubilization, absorption, and bioavailability. Liposomes can be particularly useful in terms of drug delivery due to their specificity and the long duration of action they provide.

[0082] The compositions may include pharmaceutically acceptable salts, esters, or salts of such esters, or other compounds that, upon administration to a subject, can provide (directly or indirectly) a biologically active metabolite or residue of the related compound (e.g., MANP). Thus, in one aspect, the present disclosure provides pharmaceutically acceptable salts of MANPs, prodrugs and pharmaceutically acceptable salts of such prodrugs, as well as other bioequivalents. Prodrugs are therapeutic agents that are prepared in an inactive form and are converted to an active form (i.e., a drug) within the body or cells by the action of endogenous enzymes or other chemicals and / or conditions. The term "pharmaceutically acceptable salt" refers to physiologically and pharmaceutically acceptable salts of MANPs useful in the methods provided herein (i.e., salts that retain the desired biological activity of the parent compound without imparting undesirable toxic effects). Examples of pharmaceutically acceptable salts include, but are not limited to, salts formed with cations (e.g., sodium, potassium, calcium, or polyamines such as spermine), acid addition salts formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, or nitric acid), salts formed with organic acids (e.g., acetic acid, citric acid, oxalic acid, palmitic acid, or fumaric acid), and salts formed with elemental anions (e.g., bromine, iodine, or chlorine).

[0083] The composition may further comprise other auxiliary components that are conventionally found in pharmaceutical compositions.Therefore, in one embodiment, the composition may also comprise compatible pharmaceutically active materials, such as antipruritics, astringents, local anesthetics, or anti-inflammatory agents, or additional materials that are useful for physically formulating various dosage forms of the composition, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickeners, and stabilizers.In addition, the composition may be mixed with auxiliary agents, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for affecting osmotic pressure, buffers, coloring agents, flavoring agents, penetration enhancers, and aromatic substances.However, when adding such substances, they should not excessively inhibit the biological activity of other components in the composition.

[0084] In one embodiment, the MANPs may be formulated as sustained-release dosage forms. In one embodiment, the MANPs may be formulated in controlled-release formulations. In one embodiment, coatings, envelopes, or protective matrices may be formulated to include one or more of the MANPs described herein. Such coatings, envelopes, or protective matrices may be used to coat indwelling devices such as stents, catheters, and peritoneal dialysis tubing. In one embodiment, the polypeptides may be incorporated into polymeric substances, liposomes, microemulsions, microparticles, nanoparticles, or waxes.

[0085] The pharmaceutical preparations disclosed herein can be conveniently provided in unit dosage form, but can be prepared according to any suitable method, including conventional techniques known in the pharmaceutical industry.Such techniques include combining active ingredient with desired pharmaceutical carrier.Generally, preparations can be prepared by uniformly and intimately mixing active ingredient with liquid carrier, or finely divided solid carrier, or both, and then optionally shaping the product.If necessary, the preparation can be sterilized, as long as the sterilization method does not affect the effectiveness of the molecules contained in the preparation.

[0086] In one embodiment, MANPs may be formulated for subcutaneous delivery via injection, depot polymers, drug patches, pumps, or microparticles / nanoparticles. By way of example and not limitation, PCT Publication No. WO 2008 / 061355 discloses materials and methods for formulating polypeptides for delivery in hydrogel tubes. The polypeptide may be mixed with one or more pharmaceutically acceptable excipients that are pharmaceutically acceptable and compatible with the polypeptide in amounts suitable for use in the methods described herein. In one embodiment, the polypeptide may 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 excipient may be a solid, semi-solid, or liquid substance that acts as a vehicle, carrier, or medium for the polypeptide. In one embodiment, the polypeptide may be compressed, compacted, or extruded with one or more excipients and then inserted into a hydrogel tube. Such formulations may result in pharmaceutical compositions with desirable release characteristics, improved stability, and / or other desirable properties.

[0087] Pharmaceutical compositions may also include auxiliary agents or excipients, such as lubricants, solubilizers, surfactants, diluents, binders, disintegrants, and / or glidants. In one embodiment, the solubilizer may increase the dissolution rate of the polypeptide from the dosage formulation and, in one embodiment, may include an organic acid and / or a salt of an organic acid (e.g., sodium citrate with citric acid). Other examples of excipients useful in such formulations include synthetic, semi-synthetic, modified, and natural polymers (e.g., lactose, dextrose, sucrose, trehalose, sorbitol, mannitol, starch, acacia gum, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, PEG, cyclodextrin, alkoxy-modified cyclodextrin, hydroxyethyl cellulose, hydroxypropyl cellulose, microcrystalline cellulose, albumin, dextran, maltitol, xylitol, kaolin, methylcellulose). The polypeptide may also be mixed with lubricants (e.g., talc, magnesium stearate, stearic acid, or mineral oil, calcium stearate, hydrogenated vegetable oils, sodium benzoate, sodium chloride, leucine carbowax, magnesium lauryl sulfate, or glyceryl monostearate), wetting agents, emulsifying and suspending agents, or preservatives (e.g., ethyl or propyl hydroxybenzoates).

[0088] Other agents that may be added to the pharmaceutical composition may alter the pH of the microenvironment upon dissolution and further contribute to establishing a therapeutically effective plasma concentration profile of the MANP compound. Such agents include salts of inorganic acids and magnesium hydroxide. Other agents that may be used include surfactants and other solubilizing agents.

[0089] Useful diluents, in one embodiment, include pharmaceutically acceptable inert fillers such as microcrystalline cellulose, lactose, sucrose, fructose, glucose, dextrose, or other sugars, dicalcium phosphate, calcium sulfate, cellulose, ethyl cellulose, cellulose derivatives, kaolin, mannitol, lactitol, maltitol, xylitol, or other sugar alcohols, dry starch, sugars, dextrin, maltodextrin, or other polysaccharides, inositol, or combinations thereof. In one embodiment, water-soluble diluents are particularly useful.

[0090] Lubricants may be used to improve the flow and compressibility of the composition components during processing. Useful lubricants, in one embodiment, include colloidal silicon dioxide (also known as colloidal silica, fumed silica, light anhydrous silicic acid, anhydrous silicic acid, and fumed silicon dioxide).

[0091] Suitable surfactants for use in the pharmaceutical compositions described herein include, but are not limited to, sodium lauryl sulfate, polyethylene stearic acid esters, polyethylene sorbitan fatty acid esters, polyoxyethylene castor oil derivatives, polyoxyethylene alkyl ethers, benzyl benzoate, cetrimide, cetyl alcohol, docusate sodium, glyceryl monooleate, glyceryl monostearate, glyceryl palmitostearate, lecithin, medium chain triglycerides, monoethanolamine, oleic acid, poloxamer, polyvinyl alcohol, and sorbitan fatty acid esters.

[0092] Suitable disintegrants, in one embodiment, include starch, sodium starch glycolate, crospovidone, croscarmellose, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, pectin, potassium methacrylate-divinylbenzene copolymer, polyvinyl alcohol), ethylamide, sodium bicarbonate, sodium carbonate, starch derivatives, dextrin, beta cyclodextrin, dextrin derivatives, magnesium oxide, clay, bentonite, and combinations thereof.

[0093] In one embodiment, MANPs can be incorporated into a hydrogel delivery system. In one embodiment, MANPs can be formulated for subcutaneous delivery to a patient via a xerogel-hydrogel system that can continuously and sustainably release polypeptides over an extended period of time. See, for example, U.S. Patent No. 5,226,325 and PCT Publication No. WO2004 / 071736, both of which are incorporated by reference in their entirety.

[0094] Liquid polymerizable materials useful for preparing hydrogel tubes include a wide variety of polymerizable hydrophilic and ethylenically unsaturated compounds. See, for example, the compounds described in PCT Publication No. WO 2008 / 061355. A mixture of such hydrophilic monomers is typically used in the polymerization reaction. The types and proportions of the monomers are selected to provide, upon hydration, a polymer (e.g., a crosslinked homogeneous polymer) with the desired properties (e.g., equilibrium water content (EWC) value and / or pore size) for the intended application or use.

[0095] In one embodiment, polymerization of a hydrophilic monomer mixture can result in a homogeneous hydrophilic copolymer that dissolves to varying degrees in aqueous media. In such cases, a small amount (e.g., up to about 3%) of a copolymerizable polyethylenically unsaturated crosslinker can be included in the monomer mixture to obtain a homogeneous crosslinked copolymer that is both water-insoluble and water-swellable. A slightly crosslinked homopolymer of (hydroxyethyl)methacrylate (HEMA) has an EWC value of about 38%. Crosslinked copolymers of HEMA and N-(2-hydroxypropyl)methacrylamide (HPMA) have EWC values ​​below 38%, while crosslinked copolymers of HEMA and acrylamide have EWC values ​​above 38% w / v. Therefore, depending on the useful or effective elution rate of the polypeptide, copolymer hydrogels can be customized to elute the polypeptide at the desired rate. Typically, copolymers contain about 15 to about 70% by weight of HEMA units and about 85 to about 30% by weight of a second ethylenic monomer, thus having an EWC value ranging from about 20% to about 75%. In one embodiment, the copolymer mixture may further include a small amount of a polyethylenically unsaturated crosslinker (eg, ethylene glycol dimethacrylate ("EDMA") or trimethylolpropane trimethacrylate ("TMPTMA")).

[0096] In one embodiment, a pharmaceutical composition for controlled-release delivery of MANPs in a subject may comprise (a) a complex of a polypeptide (the polypeptide having at least one basic functional group) and a polyanion derived from hexahydroxycyclohexane (the polyanion having at least two negatively charged functional groups), and (b) a pharmaceutically acceptable carrier comprising a biodegradable, water-insoluble polymer. Such compositions are described, for example, in PCT Publication No. WO 2006 / 017852 and may be prepared in the form of a solution, suspension, dispersion, emulsion, drop, aerosol, cream, semisolid, paste, capsule, tablet, solid implant, or microparticle. As used herein, the term "controlled-release delivery" refers to the continuous delivery of a pharmaceutical agent in vivo over a period of time (e.g., days to weeks or months) after administration. Sustained controlled-release delivery of MANPs, in one embodiment, may be demonstrated by the continued therapeutic effect of the polypeptide over a prolonged period (e.g., sustained relief of symptoms over a prolonged period). Sustained delivery of the polypeptide may also be demonstrated by detecting the presence of the polypeptide in vivo over time. The composition provides a low initial burst delivery followed by a steady, controlled release of the polypeptide in vivo over an extended period (eg, days to months).

[0097] In one embodiment, when a polypeptide and a polyanion are appropriately combined, a physically and chemically stable complex can be formed. The complex may take the form of a precipitate formed when an aqueous preparation of the polypeptide and polyanion is combined. Optionally, one or more pharmaceutically acceptable excipients may be incorporated into the complex. Such excipients may function 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.

[0098] The stable complex of the polypeptide and polyanion may be incorporated into a pharmaceutically acceptable carrier comprising a biodegradable, water-insoluble polymer, optionally with one or more excipients. The term "biodegradable, water-insoluble polymer" refers to biocompatible and / or biodegradable synthetic and natural polymers that may be used in vivo. The term is also intended to encompass polymers that are insoluble or become insoluble in water or biological fluids at 37°C. These polymers may be purified (e.g., to remove monomers and oligomers) using techniques well known to those skilled in the art. See, e.g., U.S. Pat. No. 4,728,721. Examples of useful polymers include, but are not limited to, polylactides, polyglycolides, poly(lactide-co-glycolides), polycaprolactones, polydioxanones, polycarbonates, polyhydroxybutyrates, polyalkylene oxalates, polyanhydrides, polyamides, polyesteramides, polyurethanes, polyacetals, polyorthocarbonates, polyphosphazenes, polyhydroxyvalerates, polyalkylene succinates, and polyorthoesters, as well as copolymers, block copolymers, branched copolymers, terpolymers, and combinations thereof.

[0099] The biodegradable, water-insoluble polymer may also include end-capped polymers, un-end-capped polymers, or mixtures of end-capped and un-end-capped polymers. Un-end-capped polymers are generally defined as having capped carboxyl end groups, while un-capped polymers have free carboxyl end groups.

[0100] Factors to consider when determining the appropriate molecular weight of a polymer include the desired rate of polymer degradation, mechanical strength, and rate of dissolution of the polymer in a solvent. A useful molecular weight of a polymer, in one embodiment, may be from about 2,000 daltons to about 150,000 daltons, with a dispersity index of 1.1 to 2.8. These may vary depending on the polymer selected for use.

[0101] Pharmaceutically acceptable carriers may be carriers that have environmentally responsive properties (e.g., heat-sensitive, pH-sensitive, or electrically sensitive) and may be in the form of an injectable solution or suspension, particles, films, pellets, cylinders, discs, microcapsules, microspheres, nanospheres, microparticles, wafers, micelles, liposomes, or any other polymeric configuration useful for drug delivery.

[0102] The method of forming various pharmaceutically acceptable polymer carriers includes those known in the art.See, for example, U.S. Patent No. 6,410,044, U.S. Patent No. 5,698,213, U.S. Patent No. 6,312,679, U.S. Patent No. 5,410,016, U.S. Patent No. 5,529,914, U.S. Patent No. 5,501,863, U.S. Patent No. 4,938,763, U.S. Patent No. 5,278,201, U.S. Patent No. 5,278,202 and PCT Publication No. WO93 / 16687.

[0103] The compositions of the present disclosure may be produced by dispersing the polypeptide / polyanion complex in a polymer matrix to form a solid implant, which may be injected or implanted into a subject. In one embodiment, such implants may be prepared using conventional polymer melt processing techniques, such as extrusion, compression molding, or injection molding. Such implants may be prepared under aseptic conditions or by terminal sterilization by irradiation (e.g., by using gamma irradiation or electron beam sterilization).

[0104] In one embodiment, compositions in the form of microspheres may be prepared by encapsulating the polypeptide / polyanion complex within a polymeric carrier using a variety of biocompatible and / or biodegradable polymers, which have properties suitable for delivery to different biological environments or for the expression of specific functions. The dissolution rate of the polypeptide, and therefore the delivery rate, is determined by factors such as the encapsulation technique, polymer composition, polymer crosslinking, polymer thickness, polymer solubility, and the size and solubility of the polypeptide / polyanion complex.

[0105] To prepare such microspheres, the polypeptide / polyanion complex to be encapsulated may be suspended in a solution of polymer in an organic solvent until the polymer solution completely coats the polypeptide / polyanion complex. This suspension may be subjected to a microencapsulation technique such as spray drying, spray congealing, emulsification, or solvent evaporation emulsification. In one embodiment, the complex or microparticles suspended with the polymer in organic solvent may be transferred to a larger volume of aqueous solution containing an emulsifier, allowing the organic solvent to evaporate or diffuse from the polymer and the solidified polymer to encapsulate the polypeptide / polyanion complex.

[0106] Emulsifiers useful for preparing encapsulated polypeptide / polyanion complexes include, for example, poloxamers and polyvinyl alcohol. Organic solvents useful in this process include acetic acid, acetone, methylene chloride, ethyl acetate, chloroform, and other non-toxic solvents depending on the properties of the polymer. Solvents are typically chosen to solubilize the polymer and are ultimately non-toxic.

[0107] In one embodiment, MANPs may be formulated into a depot formulation, which provides consistently high exposure levels and can reach high exposure levels quickly (if the lag phase is short or non-existent). See, for example, U.S. Publication No. 2010 / 0266704, the entire contents of which are incorporated herein by reference. The depot formulation may contain MANPs or a pharmaceutically acceptable salt thereof (e.g., an acid addition salt with an inorganic acid, a polymeric acid, or an organic acid). The acid addition salt may exist as a monovalent or divalent salt, depending on whether one or two equivalents of acid are added.

[0108] As described in U.S. Publication No. 2010 / 0266704, depot formulations may contain two different linear poly(lactic-co-glycolic acid) (PLGA) polymers with a molar ratio of lactic acid:glycolic acid copolymer monomers (L:G) of 85:15 to 65:35, with at least one polymer having a low intrinsic viscosity. Such formulations can maintain high plasma concentrations of the polypeptide for extended periods. Examples of suitable polymers include linear poly(D,L-lactide) and poly(D,L-lactide-co-glycolide) polymers sold under the tradenames RESOMER®, LACTEL®, and MEDISORB® by Boehringer Ingelheim Pharma GmbH & Co. KG (Ingelheim, Germany), Absorbable Polymers International (Pelham, Alabama, USA), and Alkermes, Inc. (Cambridge, Massachusetts, USA), respectively.

[0109] High exposure depot formulations for subcutaneous administration may exhibit rapid or at least very rapid action such that therapeutic plasma concentrations are achieved within a short period of time (e.g., 1, 2, 3, 4, 5, 6, or 7 days after subcutaneous injection) and may exhibit continuously high exposure levels for about a month or longer.

[0110] In one embodiment, the depot formulations provided herein may comprise two different PLGA polymers mixed or blended in a weight percent 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 embodiment, the polymer with the higher intrinsic viscosity may be present in a higher weight percent than the polymer with the lower intrinsic viscosity. In one embodiment, the polymer with the higher intrinsic viscosity may have ester end groups. The depot formulations may further contain other linear or star-shaped PLGA polymers, or polymers such as poly(D,L-lactide-co-glycolide) (PLG) or polylactic acid (PLA) polymers, provided that favorable pharmacokinetic properties are maintained.

[0111] The polypeptide content (loading amount) in the depot formulation may be in the range of 1% to 30% (e.g., 10% to 25%, more preferably 15% to 20%). The loading amount is defined as the weight ratio of the polypeptide to the total mass of the PLGA formulation.

[0112] The depot compositions may be manufactured aseptically or non-aseptically and terminally sterilized (e.g., using gamma irradiation). Terminal sterilization may provide a product with the highest possible assurance of sterility.

[0113] The depot composition may also contain one or more pharmaceutical excipients that can modify the release profile of the polypeptide. Such excipients may 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 poloxamer (also known as poly(oxyethylene-block-oxypropylene), poly(oxyethylene)-sorbitan-fatty acid esters commercially available under the trade name TWEEN®), sorbitan fatty acid esters, lecithin, inorganic salts (e.g., zinc carbonate, magnesium hydroxide, magnesium carbonate, protamine), and natural or synthetic polymers containing amine residues (e.g., polylysine).

[0114] Depot compositions may comprise a mixture or blend of polymers that differ in composition, molecular weight, and / or polymer structure.Polymer blends are defined herein as a solid solution or suspension of two different linear polymers in one implant or microparticle.Depot mixtures are defined herein as a mixture of two depot-like implants or microparticles or semi-solid formulations with different compositions, each containing one or more PLGAs.Medicinal depot compositions in which two PLGAs exist as a polymer blend can be particularly useful.

[0115] The pharmaceutical depot composition may be in the form of an implant, a semi-solid (gel), a liquid solution, a microparticle, or a suspension that solidifies in situ upon injection. The following paragraph focuses on polymeric microparticles, but is also applicable to implants, semi-solids, and liquids.

[0116] The microparticles may have a diameter of a few submicrons to several millimeters (e.g., from about 0.01 microns to about 2 mm, from about 0.1 microns to about 500 microns, from about 10 microns to about 200 microns, from about 10 microns to about 130 microns, or from about 10 microns to about 90 microns).

[0117] In one embodiment, the microparticles may be mixed with or coated with an anti-agglomerating agent. Suitable anti-agglomerating agents include, for example, mannitol, glucose, dextrose, sucrose, sodium chloride, and water-soluble polymers such as polyvinyl alcohol, polyvinylpyrrolidone, and PEG.

[0118] Microparticles may be produced using processes well known to those skilled in the art, such as by coacervation or phase separation, spray drying, oil-in-water (W / O), water-in-oil-in-water (W / O / W), or solid-oil-in-water (S / O / W) emulsification / suspension methods followed by solvent extraction or solvent evaporation. Emulsion / suspension methods are particularly useful and may include the following steps: (i) preparing an internal organic phase, (a) dissolving the polymer(s) in a suitable organic solvent (e.g., ethyl acetate, acetone, THF, acetonitrile, or halogenated hydrocarbons such as methylene chloride, chloroform, hexafluoroisopropanol, etc.) or solvent mixture, and optionally dissolving / dispersing suitable additives; (b) a step comprising dissolving / suspending / emulsifying a polypeptide in the polymer solution obtained in step (a); (ii) preparing an external aqueous phase comprising one or more stabilizers (e.g., poly(vinyl alcohol), hydroxyethyl cellulose, hydroxypropyl cellulose, poly(vinylpyrrolidone), or gelatin), and optionally a buffer salt; (iii) mixing the inner organic phase and the outer aqueous phase to form an emulsion; and (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 freeze-drying or drying under vacuum), and sieving the microparticles (e.g., through 140 μm).

[0119] The dried microparticle composition may be terminally sterilized by gamma irradiation either in bulk or after dispensing into final containers. In one embodiment, the bulk sterilized microparticles may be resuspended in a suitable vehicle, dispensed into a suitable device such as a double-chamber syringe, and then lyophilized.

[0120] In one embodiment, the microparticle depot composition may include a vehicle to facilitate reconstitution. Furthermore, prior to administration, the microparticles may be suspended in a suitable vehicle for injection (e.g., an aqueous vehicle containing one or more pharmaceutical excipients such as mannitol, sodium chloride, glucose, dextrose, sucrose, or glycerin, and / or one or more non-ionic surfactants such as poloxamer, poly(oxyethylene)-sorbitan-fatty acid esters, sodium carboxymethylcellulose, sorbitol, poly(vinylpyrrolidone), or aluminum monostearate).

[0121] How to administer MANP In one embodiment, before administering MANP or a nucleic acid encoding MANP, or one or more compositions comprising MANP to a patient, an assessment is carried out to determine whether the patient has a need for treatment for hypertension with metabolic syndrome.In one embodiment, just before administering MANP or a nucleic acid encoding MANP to a patient, an assessment is carried out to determine whether the patient has a need for treatment for hypertension with metabolic syndrome.In one embodiment, 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 before administering MANP or a nucleic acid encoding MANP to a patient, an assessment is carried out to determine whether the patient has a need for treatment for hypertension with metabolic syndrome. In one embodiment, the assessment comprises a baseline measurement of one or more parameters selected from the group consisting of plasma cGMP levels, BMI, waist circumference, serum triglyceride levels, HDL cholesterol levels, fasting plasma glucose levels, sitting systolic blood pressure, sitting diastolic blood pressure, plasma ANP-like peptide levels, insulin sensitivity, and plasma NEFA levels in accordance with the present disclosure.

[0122] In one embodiment, the patient in need of such treatment has a low baseline plasma cGMP level measured prior to receiving MANP in accordance with the present disclosure. In one embodiment, the plasma baseline cGMP level is less than 10 pmol / mL. In one embodiment, the plasma baseline cGMP level is less than 8 pmol / mL. In one embodiment, the low plasma baseline cGMP level may be 0.1-12 pmol / mL, 2-12 pmol / mL, 4-12 pmol / mL, 6-12 pmol / mL, 8-12 pmol / mL, or 10-12 pmol / mL. In one embodiment, the low plasma baseline cGMP level may be 0.1-10 pmol / mL, 0.1-8 pmol / mL, 0.1-6 pmol / mL, 0.1-4 pmol / mL, or 0.1-2 pmol / mL. In one embodiment, the low plasma baseline cGMP level may be 2-10 pmol / mL, 4-8 pmol / mL, 2-6 pmol / mL, 4-8 pmol / mL, or 6-10 pmol / mL. In one embodiment, plasma baseline plasma cGMP levels are measured using a commercially available enzyme-linked immunosorbent assay (ELISA) kit.

[0123] In one embodiment, a patient in need of such treatment receives a daily dose of 25 kg / m 2 In one embodiment, the patient in need of such treatment has a BMI of greater than 30 kg / m 2 In one embodiment, the patient in need of such treatment has a BMI of greater than 35 kg / m 2 In one embodiment, the patient in need of such treatment has a BMI of 20 to 50 kg / m 2 , 25~50kg / m 2 , 30-50kg / m 2 , 35~50kg / m 2 , 40-50kg / m 2 , or 45-50 kg / m 2 In one embodiment, the patient has a BMI of 20 to 45 kg / m 2 , 20~40kg / m 2 , 20-35kg / m 2 , or 20-30 kg / m 2In one embodiment, the patient in need of such treatment has a blood pressure of 25 to 35 kg / m 2 , 30-40kg / m 2 , 35-45kg / m 2 In one embodiment, BMI is measured by dividing a person's weight (in kilograms or pounds) by the square of their height (in meters or feet).

[0124] In one embodiment, the patient in need of such treatment is a male patient with a waist circumference of 102 cm or greater. In one embodiment, the patient in need of such treatment is a male patient with a waist circumference of 90 to 300 cm, 90 to 250 cm, 90 to 200 cm, 90 to 150 cm, or 90 to 100 cm. In one embodiment, the patient in need of such treatment is a male patient with a waist circumference of 100 to 300 cm, 150 to 300 cm, 200 to 300 cm, or 250 to 300 cm. In one embodiment, the patient in need of such treatment is a male patient with a waist circumference of 100 to 250 cm, 150 to 200 cm, 100 to 200 cm, or 150 to 250 cm.

[0125] In one embodiment, the patient in need of such treatment is a female patient with a waist circumference of 88 cm or greater. In one embodiment, the patient in need of such treatment is a female patient with a waist circumference of 80 to 300 cm, 80 to 200 cm, 80 to 150 cm, or 80 to 100 cm. In one embodiment, the patient in need of such treatment is a female patient with a waist circumference of 100 to 300 cm, 150 to 300 cm, 200 to 300 cm, or 250 to 300 cm. In one embodiment, the patient in need of such treatment is a female patient with a waist circumference of 100 to 250 cm, 150 to 200 cm, 100 to 200 cm, or 150 to 250 cm.

[0126] In one embodiment, the patient in need of such treatment has elevated serum triglyceride levels prior to receiving MANP in accordance with the present disclosure. In one embodiment, elevated serum triglyceride levels are identified by serum triglyceride levels of 150 mg / dL or greater. In one embodiment, elevated serum triglyceride levels are 100-5000 mg / dL, 100-4000 mg / dL, 100-3000 mg / dL, 100-2000 mg / dL, 100-1000 mg / dL, 100-500 mg / dL, 100-300 mg / dL, or 100-200 mg / dL. In one embodiment, the elevated serum triglyceride level is 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 another embodiment, the elevated serum triglyceride level is 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 embodiment, serum triglyceride levels are measured by a blood test after an overnight fast.

[0127] In one embodiment, the patient in need of such treatment has a low HDL cholesterol level prior to receiving MANP in accordance with the present disclosure. In one embodiment, a low HDL cholesterol level is identified by an HDL cholesterol level of less than 40 mg / dL in a male patient. In one embodiment, the patient in need of such treatment is a male patient with an HDL cholesterol level of 0.1-50 mg / dL, 0.1-40 mg / dL, 0.1-30 mg / dL, 0.1-20 mg / dL, or 0.1-10 mg / dL. In one embodiment, the patient in need of such treatment is a male patient with an HDL cholesterol level of 10-50 mg / dL, 20-50 mg / dL, 30-50 mg / dL, or 40-50 mg / dL. In one embodiment, the patient in need of such treatment is a male patient with an HDL cholesterol level of 10-40 mg / dL, 20-30 mg / dL, 10-30 mg / dL, or 20-40 mg / dL.

[0128] In one embodiment, low HDL cholesterol levels are identified by HDL cholesterol levels of less than 50 mg / dL in a female patient. In one embodiment, the patient in need of this treatment is a female patient with an HDL cholesterol level of 0.1 to 60 mg / dL, 0.1 to 50 mg / dL, 0.1 to 40 mg / dL, 0.1 to 30 mg / dL, 0.1 to 20 mg / dL, 0.1 to 10 mg / dL, 0.1 to 5 mg / dL, 0.1 to 2.5 mg / dL, or 0.1 to 1 mg / dL. In one embodiment, the patient in need of this treatment is a female patient with an HDL cholesterol level of 10 to 60 mg / dL, 20 to 60 mg / dL, 30 to 60 mg / dL, 40 to 60 mg / dL, or 50 to 60 mg / dL. In one embodiment, the patient in need of such treatment is a female patient with an HDL cholesterol level of 10-50 mg / dL, 20-40 mg / dL, 10-30 mg / dL, or 30-50 mg / dL.

[0129] In one embodiment, the patient in need of such treatment has low HDL cholesterol levels and is receiving medication to increase HDL cholesterol levels. In one embodiment, HDL cholesterol levels are measured by a blood test.

[0130] In one embodiment, the patient in need of such treatment has an elevated fasting plasma glucose level measured prior to receiving MANP in accordance with the present disclosure. In one embodiment, elevated fasting plasma glucose is identified by the patient having a fasting plasma glucose level of 100 mg / dL or greater. In one embodiment, the patient in need of such treatment has a fasting plasma glucose level between 80-200 mg / dL, between 80-180 mg / dL, between 80-160 mg / dL, between 80-140 mg / dL, between 80-120 mg / dL, or between 80-100 mg / dL. In one embodiment, the patient in need of such treatment has a fasting plasma glucose level of 100-200 mg / dL, 120-200 mg / dL, 140-200 mg / dL, 160-200 mg / dL, or 180-200 mg / dL. In one embodiment, the patient in need of such treatment has a fasting plasma glucose level of 100-180 mg / dL, 120-160 mg / dL, or 100-140 mg / dL. In one embodiment, the patient in need of such treatment is receiving medication for elevated blood glucose levels to lower blood glucose levels. In one embodiment, fasting plasma glucose levels are measured after an overnight fast using a glucose reagent from Roche (Indianapolis, IN).

[0131] In one embodiment, the patient in need of such treatment exhibits 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 one embodiment, the patient in need of such treatment exhibits hypertension despite use of 1 to 10, 1 to 8, 1 to 6, 1 to 4, 1 to 2, 2 to 10, 4 to 10, 6 to 10, 8 to 10, 2 to 8, 4 to 6, 1 to 3, 2 to 4, 3 to 5, 6 to 8, 7 to 9, or 9 to 10 antihypertensive drugs. In one embodiment, the antihypertensive drug is selected from the group consisting of a diuretic, angiotensin-converting enzyme (ACE) inhibitor, angiotensin II receptor blocker (ARB), calcium channel blocker, beta-blocker, renin inhibitor, alpha-blocker, alpha-beta-blocker, centrally acting drug, vasodilator, aldosterone antagonist, and combinations thereof.

[0132] In one embodiment, the patient in need of such treatment does not have a known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds. In one embodiment, the patient in need of such treatment is not pregnant. In one embodiment, the patient in need of such treatment is not breastfeeding. In one embodiment, the patient in need of such treatment does not have a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery. In one embodiment, the patient in need of such treatment does not have a history of severe allergies. In one embodiment, the patient in need of such treatment has not taken a phosphodiesterase 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to administering MANP in accordance with the present disclosure.

[0133] In one embodiment, the patient in need of such treatment does not exhibit any characteristic selected from the group consisting of: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; being pregnant or lactating; having a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and taking a phosphodiesterase 5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to said administration.

[0134] In one embodiment, the patient in need of such treatment does not have a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or drug abuse or addiction (within two years prior to receiving MANPs according to the methods of the present disclosure). In one embodiment, the patient in need of such treatment does not have a history of difficulty donating blood or have not donated blood or blood products within 45 days prior to administration. In one embodiment, the patient in need of such treatment does not have a history of coronary artery disease, cerebrovascular disease, or syncope. In one embodiment, the patient in need of such treatment does not have a history of epilepsy or other seizure disorders. In one embodiment, the patient in need of such treatment has not had an organ transplant. In one embodiment, the patient in need of such treatment does not have a malignancy within five years prior to receiving MANPs according to the methods of the present disclosure. In one embodiment, malignancy refers to any type of malignancy, including, but not limited to, bladder cancer, breast cancer, colorectal cancer, endometrial cancer, renal cancer, leukemia, osteosarcoma, chordoma, glioblastoma, astrocytoma, and skin cancer.

[0135] After identifying a patient as needing treatment for hypertension with metabolic syndrome, the patient can be treated with the compositions described herein according to the methods described herein. In one embodiment, a composition comprising MANPs can be administered to a patient in need of such treatment in any amount, at any frequency, and for any duration effective to achieve the desired result (e.g., to lower the patient's blood pressure or to prevent or delay a further increase in the patient's blood pressure).

[0136] In one embodiment, a method for treating hypertension with metabolic syndrome, administered to a patient in need thereof who has low baseline plasma cGMP levels, comprises parenterally administering to the patient an effective amount of MANP to increase plasma cGMP levels.

[0137] In one embodiment, a method for treating hypertension with metabolic syndrome in a patient in need thereof and having a baseline plasma cGMP level below 10 pmol / mL comprises parenterally administering to the patient an effective amount of MANP. In one embodiment, a method for treating hypertension with metabolic syndrome in a patient in need thereof and having a baseline plasma cGMP level below 8 pmol / mL comprises parenterally administering to the patient an effective amount of MANP.

[0138] In one embodiment, the disclosed method elevates plasma cGMP levels by 2-8 pmol / mL from baseline within 0.5 hours after administration of an effective amount of MANP in a patient in need of such treatment. In one embodiment, the disclosed method elevates plasma cGMP levels by 0.1-10 pmol / mL, 0.1-8 pmol / mL, 0.1-6 pmol / mL, 0.1-4 pmol / mL, or 0.1-2 pmol / mL from baseline within 0.5 hours after administration of an effective amount of MANP. In one embodiment, the disclosed method elevates plasma cGMP levels by 2-10 pmol / mL, 4-10 pmol / mL, 6-10 pmol / mL, or 8-10 pmol / mL from baseline within 0.5 hours after administration of an effective amount of MANP. In one embodiment, the disclosed method elevates plasma cGMP levels by 2-8 pmol / mL, 4-6 pmol / mL, 2-6 pmol / mL, or 4-8 pmol / mL from baseline within 0.5 hours after administration of an effective amount of MANP.

[0139] In one embodiment, the disclosed method elevates plasma cGMP levels by 1-5 pmol / mL from baseline within 4 hours of administering an effective amount of MANP to a patient in need of such treatment. In one embodiment, the disclosed method elevates plasma cGMP levels by 0.1-6 pmol / mL, 0.1-4 pmol / mL, or 0.1-2 pmol / mL from baseline within 4 hours of administering an effective amount of MANP. In one embodiment, the disclosed method elevates plasma cGMP levels by 2-6 pmol / mL, 4-6 pmol / mL, or 2-4 pmol / mL from baseline within 4 hours of administering an effective amount of MANP.

[0140] In one embodiment, the disclosed method increases plasma cGMP levels by 0.1 to 10 pmol / mL from baseline in a patient in need of such treatment within 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 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 amount of MANP. In one embodiment, the disclosed method elevates plasma cGMP levels by 0.1 to 10 pmol / mL from baseline 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 amount of MANP in a patient in need of such treatment. In another embodiment, the disclosed method elevates plasma cGMP levels by 0.1 to 10 pmol / mL from baseline within 1 to 6 hours, 2 to 6 hours, or 4 to 6 hours after administration in a patient in need of such treatment. The time after administration may be 1 to 5 hours, 2 to 4 hours, 3 to 4 hours, 0.5 to 2 hours, or 1 to 3 hours after administration of an effective amount of MANP.

[0141] As used herein, "parenteral drug administration" refers to a drug administered by a route other than the digestive tract. Typically, the term parenteral drug is used to describe a drug administered by injection or infusion. In one embodiment, parenteral administration is selected from the group consisting of subcutaneous administration (under the skin), intramuscular administration (inside the muscle), intravenous administration (inside the vein), and intrathecal administration (around the spinal cord).

[0142] In one embodiment, a method for treating hypertension with metabolic syndrome, which is applied to a patient in need of such treatment who has a low baseline plasma cGMP level, comprises administering an effective amount of MANP to a patient in need thereof at a polypeptide amount of about 0.01 ng to about 100 mg per kg of body weight (e.g., a polypeptide amount of about 10 ng to about 50 mg per kg of body weight, a polypeptide amount of about 20 ng to about 10 mg per kg of body weight, a polypeptide amount of about 0.1 ng to about 20 ng per kg of body weight, a polypeptide amount of about 3 ng to about 10 ng per kg of body weight, a polypeptide amount of about 10 ng to about 50 mg ... The dosage regimen includes parenteral administration at a dose of about 0 ng of polypeptide, about 50 ng to about 100 μg of polypeptide per kg of body weight, about 1 μg to about 10 μg of polypeptide per kg of body weight, about 1 μg to about 5 μg of polypeptide per kg of body weight, about 3 μg to about 7 μg of polypeptide per kg of body weight, about 5 μg to about 10 μg of polypeptide per kg of body weight, about 10 μg to about 20 μg of polypeptide per kg of body weight, or about 20 μg to about 100 μg of polypeptide per kg of body weight, although other dosage regimens may also provide beneficial results. In one embodiment, the parenteral administration is subcutaneous administration. In one embodiment, a composition comprising MANP may be administered parenterally (e.g., by injection, such as subcutaneous injection, or by oral ingestion) at a dose of about 0.5 ng to about 20 g of polypeptide (e.g., about 500 ng to about 10 g of polypeptide, about 1 μg to about 2 g of polypeptide, about 5 ng to about 4 μg of polypeptide, about 150 ng to about 2 μg of polypeptide, about 0.5 μg to about 10 μg of polypeptide, about 2.5 μg to about 20 mg of polypeptide, about 50 μg to about 2 mg of polypeptide, about 50 μg to about 1 mg of polypeptide, about 150 μg to about 1.4 mg of polypeptide, about 250 μg to about 2 mg of polypeptide, about 0.5 mg to about 4 mg of polypeptide, or about 1 mg to about 20 mg of polypeptide), although other dosages may also provide beneficial results.In one embodiment, MANP may be administered by infusion at a dose of, for example, about 1 pmol / kg / min to about 500 nmol / kg / min (e.g., about 1-10 pmol / kg / min, about 10-100 pmol / kg / min, about 100-300 pmol / kg / min, about 250-500 pmol / kg / min, about 500 pmol / kg / min to about 1 nmol / kg / min, about 1-10 nmol / kg / min, about 10-100 nmol / kg / min, about 100-300 nmol / kg / min, or about 250-500 nmol / kg / min).

[0143] In one embodiment, a method for lowering blood pressure in a patient in need of such treatment who has a low baseline plasma cGMP level comprises parenterally administering an effective amount of MANP to the patient, thereby increasing the plasma cGMP level relative to baseline. In one embodiment, a method for lowering blood pressure in a patient in need of such treatment reduces sitting systolic blood pressure by 0.1 to 15% relative to baseline sitting systolic blood pressure within 12 hours after administration of an effective amount of MANP. In one embodiment, a method for lowering blood pressure in a patient in need of such treatment reduces sitting systolic blood pressure by 0.1 to 20%, 0.1 to 15%, 0.1 to 10%, 0.1 to 5%, 0.1 to 2.5%, or 0.1 to 1% relative to baseline sitting systolic blood pressure within 12 hours after administration of an effective amount of MANP. In one embodiment, the method for reducing blood pressure administered to a patient in need of such treatment reduces sitting systolic blood pressure by 5-20%, 10-20%, or 15-20% compared to baseline sitting systolic blood pressure within 12 hours after administration of an effective amount of MANP. In one embodiment, the method for reducing blood pressure administered to a patient in need of such treatment reduces sitting systolic blood pressure by 5-15%, or 10-15% compared to baseline sitting systolic blood pressure within 12 hours after administration of an effective amount of MANP.

[0144] In one embodiment, the method of lowering blood pressure, administered to a patient in need of such treatment, reduces sitting systolic blood pressure by 0.1 to 20% from baseline sitting systolic blood pressure within 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 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 amount of MANP. In one embodiment, the disclosed method reduces sitting systolic blood pressure by 0.1 to 20% from baseline sitting systolic blood pressure in a patient in need of such treatment within 0.1 to 15 hours, 0.1 to 12 hours, 0.1 to 9 hours, 0.1 to 6 hours, or 0.1 to 3 hours after administration of an effective amount of MANP. In one embodiment, the disclosed method reduces sitting systolic blood pressure by 0.1 to 20% from baseline sitting systolic blood pressure in a patient in need of such treatment within 3 to 15 hours, 6 to 15 hours, 9 to 15 hours, or 12 to 15 hours after administration of an effective amount of MANP. In one embodiment, the disclosed method reduces sitting systolic blood pressure by 0.1 to 20% from baseline sitting systolic blood pressure in a patient in need of such treatment within 3 to 12 hours, 6 to 9 hours, 3 to 9 hours, or 6 to 12 hours after administration of an effective amount of MANP.

[0145] In one embodiment, a method for lowering blood pressure applied to a patient in need of such treatment reduces sitting systolic blood pressure by 0.1 to 30 mmHg compared to baseline sitting systolic blood pressure within 12 hours after administration of an effective amount of MANP. In one embodiment, a method for lowering blood pressure applied to a patient in need of such treatment reduces systolic blood pressure by 0.1 to 40 mmHg, 0.1 to 30 mmHg, 0.1 to 20 mmHg, 0.1 to 10 mmHg, 0.1 to 5 mmHg, 0.1 to 2.5 mmHg, or 0.1 to 1 mmHg compared to baseline sitting systolic blood pressure within 12 hours after administration of an effective amount of MANP. In one embodiment, a method for lowering blood pressure applied to a patient in need of such treatment reduces systolic blood pressure by 10 to 40 mmHg, 20 to 40 mmHg, or 30 to 40 mmHg compared to baseline sitting systolic blood pressure within 12 hours after administration of an effective amount of MANP. In one embodiment, the method of lowering blood pressure administered to a patient in need of such treatment reduces systolic blood pressure by 10-30 mmHg, 10-20 mmHg, 20-30 mmHg, or 15-25 mmHg relative to baseline sitting systolic blood pressure within 12 hours after administration of an effective amount of MANP.

[0146] In one embodiment, the method of lowering blood pressure administered to a patient in need of such treatment reduces sitting systolic blood pressure in the patient in need of such treatment by 0.1 to 40 mmHg from baseline systolic blood pressure within 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 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 amount of MANP. In one embodiment, the method for lowering blood pressure administered to a patient in need of such treatment reduces sitting systolic blood pressure by 0.1 to 40 mmHg from baseline systolic blood pressure in the patient in need of such treatment within 0.1 to 15 hours, 0.1 to 12 hours, 0.1 to 9 hours, 0.1 to 6 hours, or 0.1 to 3 hours after administration of an effective amount of MANP. In one embodiment, the method for lowering blood pressure administered to a patient in need of such treatment reduces sitting systolic blood pressure by 0.1 to 40 mmHg from baseline systolic blood pressure in the patient in need of such treatment within 3 to 15 hours, 6 to 15 hours, 9 to 15 hours, or 12 to 15 hours after administration of an effective amount of MANP. In one embodiment, the method for lowering blood pressure administered to a patient in need of such treatment reduces sitting systolic blood pressure by 0.1 to 40 mmHg from baseline systolic blood pressure in the patient in need of such treatment within 3 to 12 hours, 6 to 9 hours, 3 to 9 hours, or 6 to 12 hours after administration of an effective amount of MANP.

[0147] In one embodiment, a method for lowering blood pressure administered to a patient in need of such treatment reduces sitting diastolic blood pressure by 0.1 to 15% compared to baseline sitting diastolic blood pressure within 6 hours after administration of an effective amount of MANP. In one embodiment, a method for lowering blood pressure administered to a patient in need of such treatment reduces sitting diastolic blood pressure by 0.1 to 20%, 0.1 to 15%, 0.1 to 10%, 0.1 to 5%, 0.1 to 2.5 mmHg, or 0.1 to 1 mmHg compared to baseline sitting diastolic blood pressure within 6 hours after administration of an effective amount of MANP. In one embodiment, a method for lowering blood pressure administered to a patient in need of such treatment reduces sitting diastolic blood pressure by 5 to 20%, 10 to 20%, or 15 to 20% within 6 hours after administration of an effective amount of MANP. In one aspect, the method for lowering blood pressure administered to a patient in need of such treatment reduces sitting diastolic blood pressure by 5-15%, or 10-15%, within 6 hours after administration of an effective amount of MANP.

[0148] In one embodiment, the method of lowering blood pressure, administered to a patient in need of such treatment, reduces sitting diastolic blood pressure by 0.1 to 20% from baseline sitting diastolic blood pressure within 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 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 amount of MANP. In one embodiment, a method for lowering blood pressure administered to a patient in need of such treatment reduces sitting diastolic blood pressure by 0.1 to 20% from baseline sitting diastolic blood pressure within 0.1 to 8 hours, 0.1 to 7 hours, 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 amount of MANP. In one embodiment, a method for lowering blood pressure administered to a patient in need of such treatment reduces sitting diastolic blood pressure by 0.1 to 20% from baseline sitting diastolic blood pressure within 1 to 8 hours, 2 to 8 hours, 3 to 8 hours, 4 to 8 hours, 5 to 8 hours, 6 to 8 hours, or 7 to 8 hours after administration of an effective amount of MANP. In one embodiment, the method for lowering blood pressure administered to a patient in need of such treatment reduces sitting diastolic blood pressure by 0.1 to 20% from baseline sitting diastolic blood pressure within 2 to 6 hours, or 2 to 4 hours, after administration of an effective amount of MANP.

[0149] In one embodiment, a method for lowering blood pressure administered to a patient in need of such treatment reduces sitting diastolic blood pressure by 0.1 to 30 mmHg relative to baseline sitting diastolic blood pressure within 6 hours after administration of an effective amount of MANP. In one embodiment, a method for lowering blood pressure administered to a patient in need of such treatment reduces diastolic blood pressure by 0.1 to 40 mmHg, 0.1 to 30 mmHg, 0.1 to 20 mmHg, 0.1 to 10 mmHg, 0.1 to 5 mmHg, 0.1 to 2.5 mmHg, or 0.1 to 1 mmHg relative to baseline sitting diastolic blood pressure within 6 hours after administration of an effective amount of MANP. In one embodiment, a method for lowering blood pressure administered to a patient in need of such treatment reduces diastolic blood pressure by 10 to 40 mmHg, 20 to 40 mmHg, or 30 to 40 mmHg relative to baseline sitting diastolic blood pressure within 6 hours after administration of an effective amount of MANP. In one aspect, the method for lowering blood pressure administered to a patient in need of such treatment reduces diastolic blood pressure by 10-30 mmHg relative to baseline sitting diastolic blood pressure within 6 hours after administration of an effective amount of MANP.

[0150] In one embodiment, the method of lowering blood pressure administered to a patient in need of such treatment reduces sitting diastolic blood pressure in the patient in need of such treatment by 0.1 to 40 mmHg from baseline diastolic blood pressure within 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 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 amount of MANP. In one embodiment, a method for lowering blood pressure administered to a patient in need of such treatment reduces sitting diastolic blood pressure by 0.1 to 40 mmHg from baseline diastolic blood pressure in the patient in need of such treatment within 0.1 to 8 hours, 0.1 to 7 hours, 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 amount of MANP. In one embodiment, a method for lowering blood pressure administered to a patient in need of such treatment reduces sitting diastolic blood pressure by 0.1 to 40 mmHg from baseline diastolic blood pressure in the patient in need of such treatment within 1 to 8 hours, 2 to 8 hours, 3 to 8 hours, 4 to 8 hours, 5 to 8 hours, 6 to 8 hours, or 7 to 8 hours after administration of an effective amount of MANP. In one embodiment, the method for lowering blood pressure administered to a patient in need of such treatment reduces sitting diastolic blood pressure by 0.1 to 40 mmHg from baseline diastolic blood pressure in the patient in need of such treatment within 2 to 6 hours, or 2 to 4 hours, after administration of an effective amount of MANP.

[0151] In one embodiment, a method for lowering blood pressure, which is applied to a patient in need of such treatment who has a low baseline plasma cGMP level, comprises administering an effective amount of MANP to a patient in need thereof in an amount of about 0.01 ng to about 100 mg of polypeptide per kg of body weight (e.g., about 10 ng to about 50 mg of polypeptide per kg of body weight, about 20 ng to about 10 mg of polypeptide per kg of body weight, about 0.1 ng to about 20 ng of polypeptide per kg of body weight, about 3 ng to about 10 ng of polypeptide per kg of body weight, about 10 ng to about 50 ng of polypeptide per kg of body weight). These include parenteral administration at a dose of about 50 ng to about 100 μg of polypeptide per kg of body weight, about 1 μg to about 10 μg of polypeptide per kg of body weight, about 1 μg to about 5 μg of polypeptide per kg of body weight, about 3 μg to about 7 μg of polypeptide per kg of body weight, about 5 μg to about 10 μg of polypeptide per kg of body weight, about 10 μg to about 20 μg of polypeptide per kg of body weight, or about 20 μg to about 100 μg of polypeptide per kg of body weight, although other dosages may also produce beneficial results. In one embodiment, a composition comprising MANP may be administered parenterally (e.g., by injection, such as subcutaneous injection, or by oral ingestion) at a dose of about 0.5 ng to about 20 g of polypeptide (e.g., about 500 ng to about 10 g of polypeptide, about 1 μg to about 2 g of polypeptide, about 5 ng to about 4 μg of polypeptide, about 150 ng to about 2 μg of polypeptide, about 0.5 μg to about 10 μg of polypeptide, about 2.5 μg to about 20 mg of polypeptide, about 50 μg to about 2 mg of polypeptide, about 50 μg to about 1 mg of polypeptide, about 150 μg to about 1.4 mg of polypeptide, about 250 μg to about 2 mg of polypeptide, about 0.5 mg to about 4 mg of polypeptide, or about 1 mg to about 20 mg of polypeptide), although other dosages may also provide beneficial results.In one embodiment, MANP may be administered by infusion at a dose of, for example, about 1 pmol / kg / min to about 500 nmol / kg / min (e.g., about 1-10 pmol / kg / min, about 10-100 pmol / kg / min, about 100-300 pmol / kg / min, about 250-500 pmol / kg / min, about 500 pmol / kg / min to about 1 nmol / kg / min, about 1-10 nmol / kg / min, about 10-100 nmol / kg / min, about 100-300 nmol / kg / min, or about 250-500 nmol / kg / min).

[0152] In one embodiment, a method for increasing plasma ANP-like peptide levels in a patient in need of such treatment who has a low baseline plasma cGMP level comprises parenterally administering to the patient an effective amount of MANP to increase the plasma cGMP level relative to baseline. In one embodiment, a method for increasing plasma ANP-like peptide levels in a patient in need of such treatment increases the plasma ANP-like peptide level by 1-70 pg / mL relative to the baseline plasma ANP-like peptide level within 0.5 hours after administration of the effective amount of MANP. In one embodiment, a method of increasing plasma ANP-like peptide levels, applied to a patient in need of such treatment, increases plasma ANP-like peptide levels by 0.1 to 100 pg / mL, 0.1 to 90 pg / mL, 0.1 to 80 pg / mL, 0.1 to 70 pg / mL, 0.1 to 60 pg / mL, 0.1 to 50 pg / mL, 0.1 to 40 pg / mL, 0.1 to 30 pg / mL, 0.1 to 20 pg / mL, 0.1 to 10 pg / mL, 0.1 to 5 pg / mL, 0.1 to 2.5 pg / mL, or 0.1 to 1 pg / mL relative to baseline plasma ANP-like peptide levels within 0.5 hours after administration of an effective amount of MANP. In one embodiment, the method of increasing plasma ANP-like peptide levels, applied to a patient in need of such treatment, increases plasma ANP-like peptide levels by 10-100 pg / mL, 20-100 pg / mL, 30-100 pg / mL, 40-100 pg / mL, 50-100 pg / mL, 60-100 pg / mL, 70-100 pg / mL, 80-100 pg / mL, or 90-100 pg / mL relative to baseline plasma ANP-like peptide levels within 0.5 hours after administration of an effective amount of MANP. In one embodiment, a method of increasing plasma ANP-like peptide levels, applied to a patient in need of such treatment, increases plasma ANP-like peptide levels by 10-90 pg / mL, 20-80 pg / mL, 30-70 pg / mL, 40-60 pg / mL, 10-30 pg / mL, 20-40 pg / mL, 30-50 pg / mL, 40-60 pg / mL, 50-70 pg / mL, 60-80 pg / mL, or 70-90 pg / mL relative to baseline plasma ANP-like peptide levels within 0.5 hours after administration of an effective amount of MANP.

[0153] In one embodiment, a method of increasing plasma ANP-like peptide levels, administered to a patient in need of such treatment, increases plasma ANP-like peptide levels by 0.1 to 100 pg / mL from baseline plasma ANP-like peptide levels within 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 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 amount of MANP. In one embodiment, a method for increasing plasma ANP-like peptide levels applied to a patient in need of such treatment increases plasma ANP-like peptide levels by 0.1 to 100 pg / mL from baseline plasma ANP-like peptide levels within 0.1 to 2 hours, 0.1 to 1.5 hours, or 0.1 to 1 hour after administration of an effective amount of MANP. In one embodiment, a method for increasing plasma ANP-like peptide levels applied to a patient in need of such treatment increases plasma ANP-like peptide levels by 0.1 to 100 pg / mL from baseline plasma ANP-like peptide levels within 0.5 to 2 hours, 1 to 2 hours, or 1.5 to 2 hours after administration of an effective amount of MANP. In one embodiment, a method for increasing plasma ANP-like peptide levels applied to a patient in need of such treatment increases plasma ANP-like peptide levels by 0.1 to 100 pg / mL from baseline plasma ANP-like peptide levels within 0.5 to 1.5 hours after administration of an effective amount of MANP.

[0154] In one embodiment, a method for increasing plasma ANP-like peptide levels applied to a patient in need of such treatment increases plasma ANP-like peptide levels by 1-40 pg / mL relative to baseline plasma ANP-like peptide levels within one hour after administration of an effective amount of MANP. In another embodiment, a method for increasing plasma ANP-like peptide levels applied to a patient in need of such treatment increases plasma ANP-like peptide levels by 0.1-60 pg / mL, 0.1-50 pg / mL, 0.1-40 pg / mL, 0.1-30 pg / mL, 0.1-20 pg / mL, or 0.1-10 pg / mL relative to baseline plasma ANP-like peptide levels within one hour after administration of an effective amount of MANP. In one embodiment, the method of increasing plasma ANP-like peptide levels, applied to a patient in need of such treatment, increases plasma ANP-like peptide levels by 10-60 pg / mL, 20-60 pg / mL, 30-60 pg / mL, 40-60 pg / mL, 50-60 pg / mL, 10-30 pg / mL, 20-40 pg / mL, or 30-50 pg / mL relative to baseline plasma ANP-like peptide levels within 1 hour after administration of an effective amount of MANP.

[0155] In one embodiment, the method of increasing plasma ANP-like peptide levels, administered to a patient in need of such treatment, increases the plasma ANP-like peptide level by 0.1 to 60 pg / mL relative to baseline plasma ANP-like peptide levels within 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 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 amount of MANP. In one embodiment, a method for increasing plasma ANP-like peptide levels applied to a patient in need of such treatment increases plasma ANP-like peptide levels by 0.1 to 60 pg / mL relative to baseline plasma ANP-like peptide levels within 0.1 to 2 hours, 0.1 to 1.5 hours, or 0.1 to 1 hour after administration of an effective amount of MANP. In one embodiment, a method for increasing plasma ANP-like peptide levels applied to a patient in need of such treatment increases plasma ANP-like peptide levels by 0.1 to 60 pg / mL relative to baseline plasma ANP-like peptide levels within 0.5 to 2 hours, 1 to 2 hours, or 1.5 to 2 hours after administration of an effective amount of MANP. In one embodiment, a method for increasing plasma ANP-like peptide levels applied to a patient in need of such treatment increases plasma ANP-like peptide levels by 0.1 to 60 pg / mL relative to baseline plasma ANP-like peptide levels within 0.5 to 1.5 hours after administration of an effective amount of MANP.

[0156] In one embodiment, plasma ANP-like peptide levels are measured using an established ANP radioimmunoassay (RIA) (Mayo Clinic, Rochester, Minnesota, USA) and Phoenix Pharmaceuticals, Mountain View, Calif., USA).

[0157] In one embodiment, a method for increasing plasma ANP-like peptide levels, which is applied to a patient in need of such treatment and whose baseline plasma cGMP levels are below 10 pmol / mL, comprises administering an effective amount of MANP to a patient in need thereof at a polypeptide amount of about 0.01 ng to about 100 mg per kg of body weight (e.g., a polypeptide amount of about 10 ng to about 50 mg per kg of body weight, a polypeptide amount of about 20 ng to about 10 mg per kg of body weight, a polypeptide amount of about 0.1 ng to about 20 ng per kg of body weight, a polypeptide amount of about 3 ng to about 10 ng per kg of body weight, a polypeptide amount of about 10 ng to about 10 mg ... These include parenteral administration at a dose of about 100 ng to about 50 ng of polypeptide per kg of body weight, about 50 ng to about 100 μg of polypeptide per kg of body weight, about 1 μg to about 10 μg of polypeptide per kg of body weight, about 1 μg to about 5 μg of polypeptide per kg of body weight, about 3 μg to about 7 μg of polypeptide per kg of body weight, about 5 μg to about 10 μg of polypeptide per kg of body weight, about 10 μg to about 20 μg of polypeptide per kg of body weight, or about 20 μg to about 100 μg of polypeptide per kg of body weight, although other dosages may also produce beneficial results. In one embodiment, a composition comprising MANP may be administered parenterally (e.g., by injection, such as subcutaneous injection, or by oral ingestion) at a dose of about 0.5 ng to about 20 g of polypeptide (e.g., about 500 ng to about 10 g of polypeptide, about 1 μg to about 2 g of polypeptide, about 5 ng to about 4 μg of polypeptide, about 150 ng to about 2 μg of polypeptide, about 0.5 μg to about 10 μg of polypeptide, about 2.5 μg to about 20 mg of polypeptide, about 50 μg to about 2 mg of polypeptide, about 50 μg to about 1 mg of polypeptide, about 150 μg to about 1.4 mg of polypeptide, about 250 μg to about 2 mg of polypeptide, about 0.5 mg to about 4 mg of polypeptide, or about 1 mg to about 20 mg of polypeptide), although other dosages may also provide beneficial results.In one embodiment, MANP may be administered by infusion at a dose of, for example, about 1 pmol / kg / min to about 500 nmol / kg / min (e.g., about 1-10 pmol / kg / min, about 10-100 pmol / kg / min, about 100-300 pmol / kg / min, about 250-500 pmol / kg / min, about 500 pmol / kg / min to about 1 nmol / kg / min, about 1-10 nmol / kg / min, about 10-100 nmol / kg / min, about 100-300 nmol / kg / min, or about 250-500 nmol / kg / min).

[0158] In one embodiment, a method for lowering plasma glucose levels applied to a patient in need of such treatment comprises parenterally administering an effective amount of MANP to the patient. In one embodiment, a method for lowering plasma glucose levels applied to a patient in need of such treatment reduces plasma glucose levels by 1 to 17 mg / mL compared to baseline plasma glucose levels within 4 hours after administration of the effective amount of MANP. In one embodiment, a method for lowering plasma glucose levels applied to a patient in need of such treatment reduces 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 levels within 4 hours after administration of the effective amount of MANP. In one embodiment, a method for lowering plasma glucose levels applied to a patient in need of such treatment reduces 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 levels within 4 hours after administration of the effective amount of MANP. In one embodiment, the method of lowering plasma glucose levels, administered to a patient in need of such treatment, reduces plasma glucose levels by between 5 and 15 mg / mL, between 5 and 10 mg / mL, between 10 and 15 mg / mL, or between 7.5 and 12.5 mg / mL, relative to baseline plasma glucose levels, within 4 hours after administration of an effective amount of MANP.

[0159] In one embodiment, the method of lowering plasma glucose levels, administered to a patient in need of such treatment, reduces plasma glucose levels by 0.1 to 20 mg / mL relative to baseline plasma glucose levels within 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 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 amount of MANP. In one embodiment, the method for lowering plasma glucose levels applied to a patient in need of such treatment reduces plasma glucose levels by 0.1 to 20 mg / mL relative to baseline plasma glucose levels 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 amount of MANP. In one embodiment, the method for lowering plasma glucose levels applied to a patient in need of such treatment reduces plasma glucose levels by 0.1 to 20 mg / mL relative to baseline plasma glucose levels 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 amount of MANP.

[0160] In one embodiment, plasma glucose levels are measured after an overnight fast using a glucose reagent from Roche (Indianapolis, IN, USA).

[0161] In one embodiment, a method for lowering plasma glucose levels, which is administered to a patient in need of such treatment, comprises administering an effective amount of MANP to a patient in need thereof in an amount of about 0.01 ng to about 100 mg of polypeptide per kg of body weight (e.g., about 10 ng to about 50 mg of polypeptide per kg of body weight, about 20 ng to about 10 mg of polypeptide per kg of body weight, about 0.1 ng to about 20 ng of polypeptide per kg of body weight, about 3 ng to about 10 ng of polypeptide per kg of body weight, about 10 ng to about 50 ng of polypeptide per kg of body weight). These dosages include parenteral administration at a dose of about 50 ng to about 100 μg of polypeptide per kg of body weight, about 1 μg to about 10 μg of polypeptide per kg of body weight, about 1 μg to about 5 μg of polypeptide per kg of body weight, about 3 μg to about 7 μg of polypeptide per kg of body weight, about 5 μg to about 10 μg of polypeptide per kg of body weight, about 10 μg to about 20 μg of polypeptide per kg of body weight, or about 20 μg to about 100 μg of polypeptide per kg of body weight, although other dosages may also produce beneficial results. In one embodiment, a composition comprising MANP may be administered parenterally (e.g., by injection, such as subcutaneous injection, or by oral ingestion) at a dose of about 0.5 ng to about 20 g of polypeptide (e.g., about 500 ng to about 10 g of polypeptide, about 1 μg to about 2 g of polypeptide, about 5 ng to about 4 μg of polypeptide, about 150 ng to about 2 μg of polypeptide, about 0.5 μg to about 10 μg of polypeptide, about 2.5 μg to about 20 mg of polypeptide, about 50 μg to about 2 mg of polypeptide, about 50 μg to about 1 mg of polypeptide, about 150 μg to about 1.4 mg of polypeptide, about 250 μg to about 2 mg of polypeptide, about 0.5 mg to about 4 mg of polypeptide, or about 1 mg to about 20 mg of polypeptide), although other dosages may also provide beneficial results.In one embodiment, MANP may be administered by infusion at a dose of, for example, about 1 pmol / kg / min to about 500 nmol / kg / min (e.g., about 1-10 pmol / kg / min, about 10-100 pmol / kg / min, about 100-300 pmol / kg / min, about 250-500 pmol / kg / min, about 500 pmol / kg / min to about 1 nmol / kg / min, about 1-10 nmol / kg / min, about 10-100 nmol / kg / min, about 100-300 nmol / kg / min, or about 250-500 nmol / kg / min).

[0162] In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment comprises parenterally administering an effective amount of MANP to the patient. In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment increases the patient's HOMA2-insulin sensitivity (HOMA2-S) level by 0.1 to 35% relative to the patient's baseline HOMA2-S level within 4 hours after administration of the effective amount of MANP. In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment increases the patient's HOMA2-S level by 0.1 to 40%, 0.1 to 30%, 0.1 to 20%, 0.1 to 10%, 0.1 to 5%, 0.1 to 2.5%, or 0.1 to 1% relative to the patient's baseline HOMA2-S level within 4 hours after administration of the effective amount of MANP. In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment increases the patient's HOMA2-S level by 10-40%, 20-40%, or 30-40% relative to the patient's baseline HOMA2-S level within 4 hours after administration of an effective amount of MANP. In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment increases the patient's HOMA2-S level by 10-30%, 10-20%, 20-30%, or 15-25% relative to the patient's baseline HOMA2-S level within 4 hours after administration of an effective amount of MANP.

[0163] In one embodiment, a method of improving insulin sensitivity administered to a patient in need of such treatment increases the patient's HOMA2-S level by 0.1 to 40% relative to the patient's baseline HOMA2-S level within 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 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 amount of MANP. In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment increases the patient's HOMA2-S level by 0.1 to 40% relative to baseline HOMA2-S levels 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 amount of MANP. In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment increases the patient's HOMA2-S level by 0.1 to 40% relative to baseline HOMA2-S levels within 1 to 6 hours, 2 to 6 hours, 3 to 6 hours, 4 to 6 hours, 5 to 6 hours, 2 to 5 hours, 3 to 4 hours, 1 to 3 hours, 2 to 4 hours, or 3 to 5 hours after administration of an effective amount of MANP.

[0164] In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment reduces the patient's HOMA2 insulin resistance (HOMA2-IR) by 0.1 to 0.6 relative to the patient's baseline HOMA2-IR within 4 hours after administration of an effective amount of MANP. In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment reduces the patient's HOMA2-IR by 0.1 to 0.8, 0.1 to 0.6, 0.1 to 0.4, or 0.1 to 0.2 relative to the patient's baseline HOMA2-IR within 4 hours after administration of an effective amount of MANP. In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment reduces the patient's HOMA2-IR by 0.2 to 1, 0.4 to 1, 0.6 to 1, or 0.8 to 1 relative to the patient's baseline HOMA2-IR within 4 hours after administration of an effective amount of MANP. In one aspect, the method for improving insulin sensitivity administered to a patient in need of such treatment reduces the patient's HOMA2-IR value by 0.2 to 0.8, 0.4 to 0.6, 0.2 to 0.6, or 0.4 to 0.8 relative to the patient's baseline HOMA2-IR value within 4 hours after administering an effective amount of MANP.

[0165] In one embodiment, a method of improving insulin sensitivity administered to a patient in need of such treatment reduces the patient's HOMA2-IR value by 0.1 to 1 relative to the patient's baseline HOMA2-IR value within 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 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 amount of MANP. In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment reduces the patient's HOMA2-IR by 0.1 to 1 relative to a baseline HOMA2-IR value 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 amount of MANP. In one embodiment, a method for improving insulin sensitivity administered to a patient in need of such treatment reduces the patient's HOMA2-IR by 0.1 to 1 relative to a baseline HOMA2-IR value within 1 to 6 hours, 2 to 6 hours, 3 to 6 hours, 4 to 6 hours, 5 to 6 hours, 2 to 5 hours, 3 to 4 hours, 1 to 3 hours, 2 to 4 hours, or 3 to 5 hours after administration of an effective amount of MANP.

[0166] In one embodiment, HOMA2-S and HOMA2-IR values ​​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).

[0167] In one embodiment, a method for improving insulin sensitivity, which is applied to a patient in need of such treatment, comprises administering an effective amount of MANP to a patient in need thereof in an amount of about 0.01 ng to about 100 mg of polypeptide per kg of body weight (e.g., about 10 ng to about 50 mg of polypeptide per kg of body weight, about 20 ng to about 10 mg of polypeptide per kg of body weight, about 0.1 ng to about 20 ng of polypeptide per kg of body weight, about 3 ng to about 10 ng of polypeptide per kg of body weight, about 10 ng to about 50 ng of polypeptide per kg of body weight). These dosages include parenteral administration at a dose of about 50 ng to about 100 μg of polypeptide per kg of body weight, about 1 μg to about 10 μg of polypeptide per kg of body weight, about 1 μg to about 5 μg of polypeptide per kg of body weight, about 3 μg to about 7 μg of polypeptide per kg of body weight, about 5 μg to about 10 μg of polypeptide per kg of body weight, about 10 μg to about 20 μg of polypeptide per kg of body weight, or about 20 μg to about 100 μg of polypeptide per kg of body weight, although other dosages may also produce beneficial results. In one embodiment, a composition comprising MANP may be administered parenterally (e.g., by injection, such as subcutaneous injection, or by oral ingestion) at a dose of about 0.5 ng to about 20 g of polypeptide (e.g., about 500 ng to about 10 g of polypeptide, about 1 μg to about 2 g of polypeptide, about 5 ng to about 4 μg of polypeptide, about 150 ng to about 2 μg of polypeptide, about 0.5 μg to about 10 μg of polypeptide, about 2.5 μg to about 20 mg of polypeptide, about 50 μg to about 2 mg of polypeptide, about 50 μg to about 1 mg of polypeptide, about 150 μg to about 1.4 mg of polypeptide, about 250 μg to about 2 mg of polypeptide, about 0.5 mg to about 4 mg of polypeptide, or about 1 mg to about 20 mg of polypeptide), although other dosages may also provide beneficial results.In one embodiment, MANP may be administered by infusion at a dose of, for example, about 1 pmol / kg / min to about 500 nmol / kg / min (e.g., about 1-10 pmol / kg / min, about 10-100 pmol / kg / min, about 100-300 pmol / kg / min, about 250-500 pmol / kg / min, about 500 pmol / kg / min to about 1 nmol / kg / min, about 1-10 nmol / kg / min, about 10-100 nmol / kg / min, about 100-300 nmol / kg / min, or about 250-500 nmol / kg / min).

[0168] In one embodiment, a method for increasing plasma NEFA levels in a patient in need of such treatment comprises parenterally administering an effective amount of MANP to the patient. In one embodiment, a method for increasing plasma NEFA levels in a patient in need of such treatment increases plasma NEFA levels by 1 μM relative to baseline plasma NEFA levels within one hour after administration of the effective amount of MANP. In one embodiment, a method for increasing plasma NEFA levels in a patient in need of such treatment increases plasma NEFA levels by 0.01-2 μM, 0.01-1.5 μM, 0.01-0.5 μM, 0.01-0.2 μM, or 0.01-0.1 μM relative to baseline plasma NEFA levels within one hour after administration of the effective amount of MANP. In one embodiment, a method for increasing plasma NEFA levels administered to a patient in need of such treatment increases plasma NEFA levels by 0.1-2 μM, 0.2-2 μM, 0.5-2 μM, 1-2 μM, or 1.5-2 μM relative to baseline plasma NEFA levels within one hour after administration of an effective amount of MANP. In one embodiment, a method for increasing plasma NEFA levels administered to a patient in need of such treatment increases plasma NEFA levels by 0.1-1.5 μM, 0.2-1 μM, 0.5-1 μM, 0.1-0.5 μM, 0.2-1 μM, or 0.5-1.5 μM relative to baseline plasma NEFA levels within one hour after administration of an effective amount of MANP.

[0169] In one embodiment, a method for increasing plasma NEFA levels, administered to a patient in need of such treatment, increases plasma NEFA levels by 0.01 to 2 μM relative to baseline plasma NEFA levels within 0.1 hours, 0.2 hours, 0.3 hours, 0.4 hours, 0.5 hours, 0.6 hours, 0.7 hours, 0.8 hours, 0.9 hours, 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 amount of MANPs. In one embodiment, a method for increasing plasma NEFA levels applied to a patient in need of such treatment increases plasma NEFA levels by 0.01 to 2 μM relative to baseline plasma NEFA levels within 0.1 to 2 hours, 0.1 to 1.5 hours, 0.1 to 1 hour, or 0.1 to 0.5 hours after administration of an effective amount of MANP. In one embodiment, a method for increasing plasma NEFA levels applied to a patient in need of such treatment increases plasma NEFA levels by 0.01 to 2 μM relative to baseline plasma NEFA levels within 0.5 to 2 hours, 1 to 2 hours, or 1.5 to 2 hours after administration of an effective amount of MANP. In one embodiment, a method for increasing plasma NEFA levels applied to a patient in need of such treatment increases plasma NEFA levels by 0.01 to 2 μM relative to baseline plasma NEFA levels within 0.5 to 1.5 hours, 0.5 to 1 hour, or 1 to 1.5 hours after administration of an effective amount of MANP.

[0170] In one embodiment, plasma NEFA levels are determined as an index of lipolysis and are measured after an overnight fast using a commercially available kit (Zem-Bio, Inc., Durham, NC, USA) according to the manufacturer's instructions.

[0171] In one embodiment, a method for increasing plasma NEFA levels, which is applied to a patient in need of such treatment, comprises administering an effective amount of MANP to a patient in need thereof at a polypeptide amount of about 0.01 ng to about 100 mg per kg of body weight (e.g., about 10 ng to about 50 mg per kg of body weight, about 20 ng to about 10 mg per kg of body weight, about 0.1 ng to about 20 ng per kg of body weight, about 3 ng to about 10 ng per kg of body weight, about 10 ng to about 50 ng per kg of body weight). These dosages include parenteral administration at a dose of about 50 ng to about 100 μg of polypeptide per kg of body weight, about 1 μg to about 10 μg of polypeptide per kg of body weight, about 1 μg to about 5 μg of polypeptide per kg of body weight, about 3 μg to about 7 μg of polypeptide per kg of body weight, about 5 μg to about 10 μg of polypeptide per kg of body weight, about 10 μg to about 20 μg of polypeptide per kg of body weight, or about 20 μg to about 100 μg of polypeptide per kg of body weight, although other dosages may also produce beneficial results. In one embodiment, a composition comprising MANP may be administered parenterally (e.g., by injection, such as subcutaneous injection, or by oral ingestion) at a dose of about 0.5 ng to about 20 g of polypeptide (e.g., about 500 ng to about 10 g of polypeptide, about 1 μg to about 2 g of polypeptide, about 5 ng to about 4 μg of polypeptide, about 150 ng to about 2 μg of polypeptide, about 0.5 μg to about 10 μg of polypeptide, about 2.5 μg to about 20 mg of polypeptide, about 50 μg to about 2 mg of polypeptide, about 50 μg to about 1 mg of polypeptide, about 150 μg to about 1.4 mg of polypeptide, about 250 μg to about 2 mg of polypeptide, about 0.5 mg to about 4 mg of polypeptide, or about 1 mg to about 20 mg of polypeptide), although other dosages may also provide beneficial results.In one embodiment, MANP may be administered by infusion at a dose of, for example, about 1 pmol / kg / min to about 500 nmol / kg / min (e.g., about 1-10 pmol / kg / min, about 10-100 pmol / kg / min, about 100-300 pmol / kg / min, about 250-500 pmol / kg / min, about 500 pmol / kg / min to about 1 nmol / kg / min, about 1-10 nmol / kg / min, about 10-100 nmol / kg / min, about 100-300 nmol / kg / min, or about 250-500 nmol / kg / min).

[0172] In one embodiment, the disclosed method includes administering an effective amount of MANPs once (e.g., by implantation or injection of a depot composition) or multiple times (e.g., by repeated injections or oral administration). When administered multiple times, the administration frequency can range from once or more per day (e.g., once, twice, three times, four times, or more per day) to about once every two months (e.g., 3-5 times per week, about once per week, about twice per month, about once per month, or about once every two months). In one embodiment, the disclosed method includes administering MANPs via a first route (e.g., intravenously) for a first period of time, followed by administering MANPs via another route (e.g., subcutaneously) for a second period of time. In one embodiment, the methods of the disclosure include intravenously administering MANP to a patient (e.g., a human) at a dose of about 1 pmol / kg / min to about 500 nmol / kg / min for 1 hour to 7 days (e.g., 1-2 hours, 2-4 hours, 4-6 hours, 6-8 hours, 8-12 hours, 12-24 hours, 24-48 hours, 48-36 hours, 3-4 days, 4-4 days, 4-6 days, or 6-7 days), which may then be subcutaneously administered to the patient at a dose of about 0.01 ng / kg to about 100 mg / kg, 1-3 times daily for 5-30 days or more (e.g., 5-7 days, 7-10 days, 10-14 days, 14-21 days, 21-28 days, 28-30 days, or 30 days or more).

[0173] An effective amount of a MANP (or a nucleic acid encoding a MANP) administered to a patient is an amount sufficient to cause a change in a selected parameter (e.g., blood pressure) by at least 10%. In one embodiment, an effective amount of a MANP is an amount that reduces blood pressure in 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 blood pressure in said patient before administration of the MANP. In one embodiment, an effective amount of a MANP is an amount that reduces blood pressure in a patient identified as having hypertension by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 90-100%, 10-90%, 10-80%, 10-70%, 10-60%, 10-50%, 10-40%, 10-30%, 10-20%, 20-90%, 30-80%, 40-60%, 20-40%, 30-50%, 40-60%, 50-70%, 60-80%, or 70-90% compared to the blood pressure in said patient prior to administration of the MANP.

[0174] In one embodiment, an effective amount of MANP is an amount sufficient to increase plasma cGMP levels in a patient 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 plasma cGMP levels in the same patient before treatment. In one embodiment, an effective amount of MANP is an amount sufficient to increase plasma cGMP levels in a patient by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 90-100%, 10-90%, 10-80%, 10-70%, 10-60%, 10-50%, 10-40%, 10-30%, 10-20%, 20-90%, 30-80%, 40-60%, 20-40%, 30-50%, 40-60%, 50-70%, 60-80%, or 70-90% compared to the plasma cGMP level in the same patient before treatment. In one embodiment, an effective amount of MANP is an amount sufficient to increase plasma ANP-like peptide 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 plasma ANP-like peptide levels in the same patient before treatment.In one embodiment, an effective amount of MANP is an amount sufficient to increase plasma ANP-like peptide levels by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 90-100%, 10-90%, 10-80%, 10-70%, 10-60%, 10-50%, 10-40%, 10-30%, 10-20%, 20-90%, 30-80%, 40-60%, 20-40%, 30-50%, 40-60%, 50-70%, 60-80%, or 70-90% compared to plasma ANP-like peptide levels in the same patient prior to treatment. In one embodiment, an effective amount of MANP is an amount sufficient to reduce 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 plasma glucose levels in the same patient before treatment. In one embodiment, an effective amount of MANP is an amount sufficient to reduce plasma glucose levels by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 90-100%, 10-90%, 10-80%, 10-70%, 10-60%, 10-50%, 10-40%, 10-30%, 10-20%, 20-90%, 30-80%, 40-60%, 20-40%, 30-50%, 40-60%, 50-70%, 60-80%, or 70-90% compared to plasma glucose levels in the same patient before treatment. In one embodiment, an effective amount of MANP is an amount sufficient to improve 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 insulin sensitivity in the same patient before treatment.In one embodiment, an effective amount of MANP is an amount sufficient to improve insulin sensitivity by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 90-100%, 10-90%, 10-80%, 10-70%, 10-60%, 10-50%, 10-40%, 10-30%, 10-20%, 20-90%, 30-80%, 40-60%, 20-40%, 30-50%, 40-60%, 50-70%, 60-80%, or 70-90% compared to insulin sensitivity in the same patient before treatment. In one embodiment, an effective amount of MANP is an amount sufficient to increase plasma NEFA 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 plasma NEFA levels in the same patient before treatment. In one embodiment, an effective amount of MANP is an amount sufficient to increase plasma NEFA levels by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 90-100%, 10-90%, 10-80%, 10-70%, 10-60%, 10-50%, 10-40%, 10-30%, 10-20%, 20-90%, 30-80%, 40-60%, 20-40%, 30-50%, 40-60%, 50-70%, 60-80%, or 70-90% compared to plasma NEFA levels in the same patient prior to treatment.

[0175] In one embodiment, the amount and frequency of MANP administration to a patient may be titrated to identify the dose that is most effective in treating hypertension with metabolic syndrome while minimizing side effects, for example. If a patient does not respond to a particular dose, the dose may be increased, for example, two-fold, three-fold, five-fold, or ten-fold. After receiving this increased dose, the patient's response to treatment and symptoms of toxicity may both be monitored, and the dose and / or frequency of administration may be adjusted accordingly. The effective dose may remain constant or may be adjusted as a smooth step increase or decrease or variable dose depending on the patient's response to treatment.

[0176] In one embodiment, the amount and frequency of MANP administration to a patient may be titrated, for example, to identify the dose that is most effective in lowering blood pressure while minimizing side effects. If a patient does not respond to a particular dose, the dose may be increased, for example, two-fold, three-fold, five-fold, or ten-fold. After receiving this increased dose, the patient may be monitored for both responsiveness to treatment and symptoms of toxicity, and the dose and / or frequency of administration may be adjusted accordingly. The effective dose may remain constant or may be adjusted as a smooth increase / decrease or variable dose depending on the patient's response to treatment.

[0177] In one embodiment, the amount and frequency of MANP administration to a patient may be titrated, e.g., to identify the dose that is most effective in increasing plasma ANP-like peptide levels while minimizing side effects. If a patient does not respond to a particular dose, the dose may be increased, e.g., two-fold, three-fold, five-fold, or ten-fold. After receiving this increased dose, the patient may be monitored for both responsiveness to treatment and symptoms of toxicity, and the dose and / or frequency of administration may be adjusted accordingly. The effective dose may remain constant or may be adjusted as a smooth increase / decrease or variable dose depending on the patient's response to treatment.

[0178] In one embodiment, the amount and frequency of MANP administration to a patient may be titrated, for example, to identify the dose that is most effective in lowering plasma glucose levels while minimizing side effects. If a patient does not respond to a particular dose, the dose may be increased, for example, two-fold, three-fold, five-fold, or ten-fold. After receiving this increased dose, the patient may be monitored for both responsiveness to treatment and symptoms of toxicity, and the dose and / or frequency of administration may be adjusted accordingly. The effective dose may remain constant or may be adjusted as a smooth increase / decrease or variable dose depending on the patient's response to treatment.

[0179] In one embodiment, the amount and frequency of MANP administration to a patient may be titrated, for example, to identify the dose that is most effective in improving insulin sensitivity while minimizing side effects. If a patient does not respond to a particular dose, the dose may be increased, for example, two-fold, three-fold, five-fold, or ten-fold. After receiving this increased dose, the patient may be monitored for both responsiveness to treatment and symptoms of toxicity, and the dose and / or frequency of administration may be adjusted accordingly. The effective dose may remain constant or may be adjusted as a smooth increase / decrease or variable dose depending on the patient's response to treatment.

[0180] In one embodiment, the amount and frequency of MANP administration to a patient may be titrated, e.g., to identify the dose that is most effective in increasing plasma NEFA levels while minimizing side effects. If a patient does not respond to a particular dose, the dose may be increased, e.g., two-fold, three-fold, five-fold, or ten-fold. After receiving this increased dose, the patient may be monitored for both responsiveness to treatment and symptoms of toxicity, and the dose and / or frequency of administration may be adjusted accordingly. The effective dose may remain constant or may be adjusted as a smooth step increase or decrease or variable dose depending on the patient's response to treatment.

[0181] In one embodiment, the administration frequency may be any frequency that alleviates, for example, the symptoms of cardiovascular or cardiorenal disease in a patient without causing significant toxicity to the patient. In one embodiment, the administration frequency may be from about four times a day to about once every three months, or from about twice a day to about once every two months, or from about once a day to about once a month, or from about once every two days to about once a week. Furthermore, the administration frequency may be constant or may vary during the treatment period. As with the effective amount, various factors may affect the actual administration frequency used for a particular application. In one embodiment, the effective amount, the duration of treatment, the route of administration, and the severity of the renal condition may require increased or decreased administration frequency.

[0182] An effective administration period may be any period that alleviates the symptoms of hypertension or metabolic syndrome in a patient without causing significant toxicity to the patient. In one embodiment, an effective administration period may be any period that lowers blood pressure without causing significant toxicity to the patient. In one embodiment, an effective administration period may be any period that increases plasma ANP-like peptide levels without causing significant toxicity to the patient. In one embodiment, an effective administration period may be any period that decreases plasma glucose levels without causing significant toxicity to the patient. In one embodiment, an effective administration period may be any period that improves insulin sensitivity without causing significant toxicity to the patient. In one embodiment, an effective administration period may be any period that increases plasma NEFA levels without causing significant toxicity to the patient. An effective period may vary from one day to several days, weeks, months, or years. Generally, an effective period may range from several days to several months. In one embodiment, an effective period may range from about 1-2 weeks to about 36 months or longer. In one embodiment, treatment may be continued throughout an individual patient's lifetime. Several factors may affect the actual effective period used for a particular treatment regimen. In one aspect, the duration of effectiveness may vary depending on the frequency of administration, the amount administered, the route of administration, and the severity of the condition.

[0183] After administering the MANPs described herein to a patient, the patient may be monitored to determine whether the disorder has improved. In one embodiment, the patient may be monitored for several hours, days, months, or years after administering the MANPs described herein. In one embodiment, the patient's characteristics that may be monitored include systolic blood pressure, diastolic blood pressure, plasma cGMP levels, plasma ANP-like peptide levels, plasma glucose levels, insulin sensitivity, and plasma NEFA levels.

[0184] In one embodiment, the patient may be evaluated after treatment to determine whether one or more symptoms of the patient's disorder have been alleviated (e.g., whether the patient's blood pressure has decreased). If the patient does not respond to a particular dose of MANP, one or both amounts may be increased, for example, two-fold, three-fold, five-fold, or ten-fold. After receiving this increased dose, both the patient's responsiveness to treatment and symptoms of toxicity may be monitored and adjusted accordingly. The effective amount may remain constant or may be adjusted as a smooth step increase or decrease or variable dose depending on the patient's response to treatment.

[0185] The present invention is further described in the following examples, which do not limit the scope of the invention described in the claims. [Example]

[0186] Example 1. Study Design and Patients of a Study Investigating the Effect of MANP in Hypertensive Subjects with Metabolic Syndrome Test Design The study consisted of a screening visit (defined as the first visit on Day 0; see below for details), a 7-day period of dietary and fluid restriction, and medications as shown in Figure 2. A total of 22 patients with hypertension and metabolic syndrome were enrolled. Starting on Day 1, all subjects began a moderate-sodium (3.0 g / day) diet, with fluid intake restricted to 2.5 L / day. This diet was continued for 7 days until discharge. On Day 5, subjects were admitted to the clinic (Visit 2), and urinary sodium was 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 for 24 hours after MANP or placebo administration. Blood pressure and heart rate were measured, along with blood draws, at baseline and at several intervals after MANP or placebo administration. On day 7, at the end of the 24-hour observation period, subjects will undergo a physical examination and be discharged from the clinic. Subjects will be off antihypertensive medication only on days 6 and 7 and will be discharged from the clinic. Laboratory tests will be performed and evaluated 7 days after the MANP / placebo injection. Subjects will be contacted by phone to review laboratory test results and evaluate the occurrence of potential side effects after MANP / placebo administration.

[0187] patient Hypertension is defined as a systolic blood pressure of 140 to 180 mmHg or a diastolic blood pressure of 90 to 100 mmHg in a sitting position despite the use of at least one antihypertensive medication. Metabolic syndrome is defined according to the criteria of the National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ATP III). Specifically, metabolic syndrome is defined as the presence of any two of the following characteristics: 1. Abdominal obesity: Waist circumference of 102 cm (40 inches) or more for men and 88 cm (35 inches) or more for women; 2. Serum triglyceride level 150 mg / dL (1.7 mmol / L) or higher, 3. Serum high-density lipoprotein (HDL) cholesterol level is less than 40 mg / dL (1 mmol / L) for men and less than 50 mg / dL (1.3 mmol / L) for women, or you are currently receiving medical treatment for low HDL cholesterol levels. 4. Fasting plasma glucose level ≥ 100 mg / dL (5.6 mmol / L) or medication for elevated blood glucose levels.

[0188] The detailed inclusion criteria are as follows: 1. Hypertension with a systolic blood pressure of 140mmHg or more but 180mmHg or a diastolic blood pressure of 90mmHg or more but 100mmHg in a sitting position despite treatment with antihypertensive drugs. 2. Metabolic syndrome with any two of the following features: a) abdominal obesity, defined as a waist circumference of 102 cm (40 inches) or more for men and 88 cm (35 inches) or more for women; b) serum triglyceride level ≥ 150 mg / dL (1.7 mmol / L); c) Serum HDL cholesterol level less than 40 mg / dL (1 mm / L) for men and less than 50 mg / dL (1.3 mmol / L) for women, or currently receiving medication for low HDL cholesterol levels. d) fasting plasma glucose level ≥ 100 mg / dL (5.6 mmol / L) or medication for elevated blood glucose levels; 3. Ages 18 to 75 years old; 4. Stable use of antihypertensive medications for 30 days prior to the screening visit, and 5. Use of statins, ezetimibe, or their combination medications at a stable dose for 60 days prior to the screening visit.

[0189] Detailed inclusion and exclusion criteria are as follows: 1. Known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds, 2. Pregnant or lactating women, 3. Have received an investigational drug or used an investigational device within 30 days prior to study participation. 4. A history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on opioids, or substance abuse or addiction (within the past two years); 5. Have a history of difficulty donating blood, or have donated blood or blood products within 45 days prior to enrollment. 6.Having a clinically significant new illness in the opinion of the investigator within one month prior to screening, 7. History of severe allergies, 8. History of coronary artery disease, cerebrovascular disease, or syncope, 9. History of epilepsy or other seizure disorder, 10. History of organ transplantation 11. Malignant tumor within 5 years of the screening visit, 12. History of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery, and 13. Taking a phosphodiesterase 5 inhibitor (sildenafil, vardenafil, tadalafil) within 72 hours after receiving MANP.

[0190] Subjects who meet the inclusion criteria based on their electronic medical records will receive a letter by mail explaining the study and inquiring about their willingness to participate. At the screening visit, a medical history will be taken, a physical examination will be performed, and a laboratory history will be screened. Electrocardiograms, blood pressure, and heart rate will be recorded, and screening tests will be performed. Blood pressure inclusion criteria will be verified by the average of three sitting blood pressure measurements.

[0191] Randomization to double-blind treatment Random treatment assignment will be performed using randomization codes sent directly by the statistician to Mayo Research Pharmacy. A study pharmacist, blinded to data analysis, will assist in the preparation of MANP and placebo. A total of 22 patients will be randomized to receive a single subcutaneous injection of placebo or MANP.

[0192] statistical methods Patient data were compiled at baseline and after MANP or placebo injection. Data distribution was tested for normality, and no data were excluded from the analysis. All adverse events were summarized as number (percentage). Means for 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 between-group effects of placebo and MANP and to evaluate between-group differences at specific time points. Comparisons with baseline levels within a single group were also performed. Multiple comparison correction was performed using the Scheffe method. Pharmacokinetic and pharmacodynamic analyses were performed using the R package (PKNCA).

[0193] C of plasma ANP-like peptide and plasma cGMP levels max The mean change in systolic and diastolic blood pressure was determined by the maximum observed value 24 hours after treatment. The mean change in systolic and diastolic blood pressure was determined by the mean change in systolic and diastolic blood pressure occurring within 24 hours after treatment compared to baseline values. Analysis of these MANP responsiveness indices between men and women was performed using unpaired t-tests assuming unequal variances. The Spearman rank method was used to assess correlations between these MANP responsiveness indices and continuous baseline variables. All statistical analyses were performed using SAS version 9.4 and Graphpad Prism 9, with a two-sided P < 0.05 considered statistically significant.

[0194] Example 2. Safety and Tolerability of MANP Subjects recruited according to Example 1 will be followed for up to 7 days after MANP / placebo injection. Safety endpoints, including adverse events, vital sign measurements, electrocardiograms, clinical laboratory results, and other safety variables, will be monitored for each subject throughout the study.

[0195] The stopping rule is defined as when two subjects meet one or both of the following criteria: 1. The occurrence of clinically significant hypotension (defined as a decrease in clinical systolic blood pressure of 30 mmHg or more from baseline, or a decrease in sitting systolic blood pressure of less than 90 mmHg, or 5 minutes of lightheadedness, dizziness, or visual symptoms on repeated blood pressure measurements), or 2. Any other safety outcome or adverse experience that, in the opinion of the investigator, raises concerns about the safety or tolerability of the administered dose.

[0196] Clinical characteristics are summarized overall and by treatment group in Table 3. Seventeen patients were randomly assigned to MANP and five to placebo. The mean baseline systolic blood pressure for the entire study cohort was 149 ± 8 mmHg, and the mean diastolic blood pressure was 83 ± 11 mmHg; all patients were taking at least one antihypertensive medication. The mean BMI and waist circumference were 36 ± 5 kg / m², respectively. 2 and 113±9 cm, indicating that this cohort was characterized by both general and abdominal obesity. [Table 3] JPEG2026507030000005.jpg79170

[0197] As shown in Table 4, no significant changes in electrocardiogram findings were observed during the 24 hours following subcutaneous MANP / placebo injection, and no drug-related, clinically relevant changes occurred in safety laboratory parameters. All 22 patients completed treatment, and none of the discontinuation criteria were met. Table 5 summarizes all treatment-related adverse events (AEs) during the 7-day follow-up period following MANP / placebo injection. No serious adverse events were observed, and orthostatic hypotension and vasovagal syncope were observed only in two patients receiving MANP. [Table 4] [Table 5]

[0198] Example 3. Changes in plasma ANP-like peptide and cGMP levels in response to MANP administration To measure plasma ANP-like peptide and cGMP levels in the subjects of Example 1, blood was collected into EDTA tubes and centrifuged at 2,500 rpm at 4°C for 10 minutes. 1 mL of plasma was then aliquoted and stored at -80°C until assayed. The assay detects both ANP and MANP, which are defined as ANP-like peptides throughout these examples. The assay's cross-reactivity is less than 1% with NT-proANP, BNP, C-type natriuretic peptide (CNP), endothelin, and adrenomedullin. Plasma cGMP levels were measured using a commercially available ELISA kit (Enzo Life Sciences, Farmingdale, NY) according to the manufacturer's instructions. Plasma ANP-like peptide and cGMP levels were measured in samples taken at baseline, 0.5, 1, 2, 4, 6, 12, and 24 hours after administration of MANP or placebo. max indicates the highest concentration detected in plasma during the 24-hour follow-up period after MANP administration. max During the 24-hour follow-up period after MANP administration, max Indicates the time it takes to reach

[0199] As shown in Table 6 and Figure 3, in patients receiving MANP, a rapid and significant increase in plasma ANP-like peptide levels occurred 30 minutes after injection (increase from baseline: 48.9 ± 19.5 pg / mL, P = 0.03), and the increase in plasma ANP-like peptide levels persisted for 1 hour after injection (increase from baseline: 26.4 ± 12.8 pg / mL, P = 0.06). Meanwhile, in the MANP group, significant increases in plasma cGMP levels were simultaneously observed at 30 minutes (increase from baseline: 4.8 ± 2.0 pmol / mL, P = 0.02) and 1 hour (increase from baseline: 2.9 ± 1.3 pmol / mL, P = 0.03). The increase persisted for approximately 4 hours after administration. In the placebo group, no significant changes were observed in plasma ANP-like peptide or cGMP levels compared with baseline. [Table 6]

[0200] Example 4. Sitting blood pressure in response to MANP administration To measure blood pressure and heart rate in subjects in Example 1, blood pressure and heart rate are monitored continuously on the day the study drug (MANP or placebo) is administered. Sitting blood pressure and heart rate are recorded before MANP or placebo injection and at 0.5, 1, 2, 4, 6, 12, and 24 hours after MANP or placebo administration.

[0201] As shown in Figure 4, in the MANP group, sitting systolic blood pressure decreased compared to baseline, with the maximum decrease occurring 6 hours after injection (change from baseline: -5.7 ± 2.9 mmHg, P = 0.06). At 12 hours after MANP administration, systolic blood pressure remained lower than baseline. In contrast, no decrease in systolic blood pressure was observed in the placebo group. Sitting diastolic blood pressure tended to be lower than baseline 6 hours after MANP administration (change from baseline: -2.2 ± 1.2 mmHg, P = 0.09). A slight increase in heart rate was observed in both groups after 6 hours after injection, but this increase was not statistically significant. There was no difference in heart rate between the MANP and placebo groups at any time point.

[0202] Example 5. Individual responses to MANP The MANP response in the subjects of Example 1 was assessed during a 24-hour follow-up period after MANP injection by: (1) plasma ANP-like peptide levels C max , (2) plasma cGMP level C max (3) mean change in systolic blood pressure, and (4) mean change in diastolic blood pressure. These results are shown in Table 7.

[0203] Increases in plasma ANP-like peptide levels were observed in all 17 patients, and baseline ANP levels were significantly higher than the C of plasma ANP-like peptide levels after MANP administration. max Furthermore, the plasma ANP-like peptide level C maxC correlates positively with waist circumference (ρ = 0.51, P = 0.04) but not with BMI (ρ = 0.28, P = 0.28). As shown in Figures 6A and 7A, age, sex, and baseline cGMP levels significantly correlated with plasma ANP-like peptide levels. max On the other hand, of these 17 patients, only two did not have elevated circulating levels of cGMP at any of the time points. As shown in Figures 6B and 7B, the C of plasma cGMP levels max is not correlated with age, sex, BMI, waist circumference, baseline plasma ANP level, and baseline plasma cGMP level. Each circle represents one subject, with black circles representing males and white circles representing females.

[0204] A decrease in systolic blood pressure was observed in all patients, and a decrease in diastolic blood pressure was observed in 15 of 17 patients who received MANP. The mean changes in systolic and diastolic blood pressure were -3.4 (IQR: -8.0 to -3.0) mmHg and -2.4 (IQR: -3.3 to 0.7) mmHg, respectively. As shown in Figures 6C and 6D, the mean change in systolic blood pressure was more pronounced in men than in women (P = 0.008), whereas the mean change in diastolic blood pressure was similar between the two genders (P = 0.25). As shown in Figures 5A and 5B, the mean change in systolic blood pressure was positively correlated with baseline plasma cGMP levels, indicating that patients with lower baseline plasma cGMP levels experienced a greater decrease in mean systolic blood pressure over the 24 hours following MANP injection (ρ = 0.66, P = 0.005). Each circle represents one subject; filled circles represent men and open circles represent women. This significant association was further verified by repeating the same analysis using median blood pressure change (baseline cGMP value vs. median systolic blood pressure change: ρ = 0.73, P = 0.0008) as shown in Table 8 or maximum blood pressure change (baseline cGMP value vs. maximum systolic blood pressure change: ρ = 0.59, P = 0.01) as shown in Table 9. For each subject, the median blood pressure (systolic or diastolic) change was defined as the median blood pressure change at 0.5, 1, 2, 4, 6, 12, and 24 hours after MANP administration. For each subject, the maximum blood pressure (systolic or diastolic) change was defined as the most extreme blood pressure change at 0.5, 1, 2, 4, 6, 12, and 24 hours after MANP administration. The mean diastolic blood pressure change was not affected by baseline plasma ANP, baseline plasma cGMP value, age, or waist circumference, and showed a weak negative correlation trend with BMI (ρ = -0.44, P = 0.08). [Table 7] [Table 8] [Table 9]

[0205] Example 6. Metabolic effects of MANPs To measure plasma metabolic parameters, after an overnight fast, plasma metabolites including glucose, insulin, NEFA, glycerol, triglycerides, and adiponectin will be measured at baseline, 0.5, 1, 2, and 4 hours after MANP or placebo administration while subjects continue to fast.

[0206] Plasma glucose levels are measured using a Roche glucose reagent (Indianapolis, IN, USA). Plasma insulin is measured using the Roche insulin assay (Indianapolis, IN, USA), a two-site immunoassay with electrochemiluminescence immunoassay (ELCIA) detection. Plasma non-esterified fatty acids (NEFAs) and glycerol are measured as indicators of lipolysis and are measured using commercially available kits (Zem-Bio, Inc., Durham, NC, USA). Plasma triglycerides are measured by an automated enzymatic method using the Roche triglyceride reagent (Indianapolis, IN, USA). This method is referenced based on standardized methods implemented at the Centers for Disease Control and Prevention (CDC) Cardiovascular Risk Assessment Laboratory. Plasma adiponectin is measured using a commercially available kit (Millipore Sigma, Burlington, MA, USA) according to the manufacturer's protocol.

[0207] As shown in Table 10, plasma glucose levels did not change significantly in the placebo group, but a trend toward a decrease in plasma glucose levels was observed 1 hour after MANP administration (change from baseline: -3.1 ± 1.5 mg / mL, P = 0.06). Importantly, plasma glucose levels decreased significantly at both 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 at 1 and 2 hours after injection, whereas plasma insulin levels remained stable in the MANP group. The difference in insulin concentrations between the MANP and placebo groups was statistically significant at both 1 hour (P = 0.02) and 2 hours (P = 0.03) after injection. In addition, a trend toward an increase in plasma NEFA was observed in the MANP-treated group, and the increase reached statistical significance 1 hour after MANP administration (P = 0.01).Plasma glycerol, triglyceride, and adiponectin levels did not change significantly during the first 4 hours after MANP injection.

[0208] As shown in Table 11, HOMA2 values ​​were calculated as an indirect and surrogate measure for the first 4 hours after injection. 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. As shown in Figure 8A, from baseline to 4 hours after injection, there was an overall increase in HOMA2-S values ​​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%)]. Concomitantly, as shown in Figure 8B, a decrease in HOMA2-IR values ​​was observed 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)]. [Table 10] [Table 11]

[0209] Other embodiments While the present disclosure has been described in connection with its detailed description, it should be understood that the foregoing description is intended to illustrate, but not limit, the scope of the disclosure, which is defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

[0210] Embodiment 1. A method of treating hypertension with metabolic syndrome, the method being applied to a patient in need of such treatment having a baseline plasma cyclic guanosine monophosphate (cGMP) level below 10 pmol / mL, the method comprising, consisting essentially of, or consisting of administering to said patient an effective amount of alternatively spliced ​​atrial natriuretic peptide (MANP), thereby increasing the plasma cGMP level compared to baseline.

[0211] Embodiment 2. The method of embodiment 1, wherein the baseline plasma cGMP level is less than 8 pmol / mL.

[0212] Embodiment 3. The method of embodiment 1 or 2, wherein said increasing is an increase of 2 to 8 pmol / mL from the baseline plasma cGMP level within 0.5 hours after said administration.

[0213] Embodiment 4. The method of embodiment 1 or 2, wherein said increasing is an increase of 1 to 5 pmol / mL from the baseline plasma cGMP level within 0.5 to 4 hours after said administration.

[0214] Embodiment 5. The method of any one of embodiments 1 to 4, wherein the MANPs are administered subcutaneously.

[0215] Embodiment 6. The method of embodiment 5, wherein the effective amount is 0.1 μg / kg to 5 μg / kg.

[0216] Embodiment 7. The method of embodiment 6, wherein the effective amount is 2.5 μg / kg.

[0217] Embodiment 8. The method of embodiment 5, wherein the effective amount is 6.5 μg to 750 μg.

[0218] Embodiment 9. The method of embodiment 8, wherein the effective amount is 162.5 μg to 375 μg.

[0219] Embodiment 10. The method of any one of embodiments 1 to 4, wherein the MANPs are administered intravenously.

[0220] Embodiment 11. The method of embodiment 10, wherein the effective amount is 10 pmol / kg / min to 100 nmol / kg / min.

[0221] Embodiment 12 The method of any one of embodiments 1 to 4, wherein the MANPs are administered intravenously followed by subcutaneous administration.

[0222] Embodiment 13. The method of embodiment 12, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 0.1 μg / kg to 5 μg / kg.

[0223] Embodiment 14. The method of embodiment 13, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 2.5 μg / kg.

[0224] Embodiment 15. The method of embodiment 12, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 6.5 μg to 750 μg.

[0225] Embodiment 16. The method of embodiment 15, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 162.5 μg to 375 μg.

[0226] Embodiment 17. The patient is taking 25 kg / m 2 17. The method of any one of embodiments 1 to 16, wherein the patient has a higher body mass index (BMI).

[0227] Embodiment 18. The patient is taking 30 kg / m 2 18. The method of any one of embodiments 1 to 17, wherein the patient has a higher BMI.

[0228] Embodiment 19. The patient is taking 35 kg / m 2 19. The method of any one of embodiments 1 to 18, wherein the patient has a higher BMI.

[0229] Embodiment 20. The method of any one of embodiments 1 to 19, wherein the patient exhibits at least two characteristics selected from the group consisting of abdominal obesity, high serum triglyceride levels, low serum high-density lipoprotein (HDL) cholesterol levels, and high fasting plasma glucose levels.

[0230] Embodiment 21. The method of embodiment 20, wherein the abdominal obesity is identified by a waist circumference of 102 cm or greater in male patients and 88 cm or greater in female patients.

[0231] Embodiment 22. The method of embodiment 20 or 21, wherein the elevated serum triglyceride level is identified by a serum triglyceride level of 150 mg / dL or greater in the patient prior to the administering.

[0232] Embodiment 23. The method of any one of embodiments 20 to 22, wherein the low HDL cholesterol level is identified by an HDL cholesterol level of less than 40 mg / dL in a male patient or an HDL cholesterol level of less than 50 mg / dL in a female patient.

[0233] Embodiment 24. The method of any one of embodiments 20 to 22, wherein the low HDL cholesterol level is identified in the patient receiving medication to increase HDL cholesterol levels for that low HDL cholesterol level.

[0234] Embodiment 25. The method of any one of embodiments 20-24, wherein the high fasting plasma glucose level is identified in the patient by a fasting plasma glucose level of 100 mg / dL or greater.

[0235] Embodiment 26. The method of any one of embodiments 20 to 24, wherein the high fasting plasma glucose level is identified in the patient receiving medication for elevated blood glucose levels to lower blood glucose levels.

[0236] Embodiment 27. The method of any one of embodiments 1 to 26, wherein the patient exhibits a sitting systolic blood pressure of 140 mmHg to 180 mmHg or a sitting diastolic blood pressure of 90 mmHg to 100 mmHg prior to said administering.

[0237] Embodiment 28. The method of any one of embodiments 1 to 27, wherein the patient has been taking an antihypertensive drug for 30 days prior to said administering, or has been taking a statin, ezetimibe, or a combination thereof for 60 days prior to said administering.

[0238] Embodiment 29. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; Taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration Having received an investigational drug or used an investigational device within 30 days prior to the administration of the investigational drug. a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or substance abuse or addiction (within two years prior to said administration); Having a history of difficulty in donating blood, or having donated blood or blood products within 45 days prior to the administration. The patient has had a clinically significant new illness within one month prior to the administration of the drug. A history of severe allergies coronary artery disease, cerebrovascular disease, or a history of syncope; a history of epilepsy or other seizure disorder; Having had an organ transplant; and Having had a malignant tumor within 5 years prior to the administration of the above. 29. The method of any one of embodiments 1 to 28, wherein the method does not exhibit any feature selected from the group consisting of:

[0239] Embodiment 30. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, History of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and Taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration 30. The method of any one of embodiments 1 to 29, wherein the method does not exhibit any feature selected from the group consisting of:

[0240] Embodiment 31. The method of any one of embodiments 1 to 30, wherein the patient's sitting systolic blood pressure is reduced by 0.1 to 15% relative to baseline sitting systolic blood pressure within 12 hours after said administration.

[0241] Embodiment 32. The method of any one of embodiments 1 to 31, wherein the patient's sitting diastolic blood pressure is reduced by 0.1 to 5% relative to baseline sitting diastolic blood pressure within 6 hours after the administration.

[0242] Embodiment 33. The method of any one of embodiments 1 to 32, wherein the patient's plasma glucose level is reduced by 1 to 17 mg / mL relative to the baseline plasma glucose level within 4 hours after the administration.

[0243] Embodiment 34. The method of any one of embodiments 1 to 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).

[0244] Embodiment 35. A method of lowering blood pressure, applied to a patient in need of such treatment having a baseline plasma cyclic guanosine monophosphate (cGMP) level below 10 pmol / mL, comprising, or consisting essentially of, or consisting of, administering to said patient an effective amount of alternatively spliced ​​atrial natriuretic peptide (MANP), thereby increasing the plasma cGMP level relative to said baseline.

[0245] Embodiment 36. The method of embodiment 35, wherein the baseline plasma cGMP level is less than 8 pmol / mL.

[0246] Embodiment 37. The method of embodiment 35 or 36, wherein said increasing is an increase of 2 to 8 pmol / mL from the baseline plasma cGMP level within 0.5 hours after said administration.

[0247] Embodiment 38. The method of embodiment 35 or 36, wherein the increasing is an increase of 1 to 5 pmol / mL from the baseline plasma cGMP level within 0.5 to 4 hours after the administration.

[0248] Embodiment 39. The method of any one of embodiments 35 to 38, wherein the reducing is a 0.1 to 15% decrease in the patient's sitting systolic blood pressure relative to baseline sitting systolic blood pressure within 12 hours after the administration.

[0249] Embodiment 40. The method of any one of embodiments 35 to 38, wherein the reducing is a 0.1 to 5% decrease in the patient's sitting diastolic blood pressure relative to baseline sitting diastolic blood pressure within 6 hours after the administration.

[0250] Embodiment 41 The method of any one of embodiments 35 to 40, wherein the MANPs are administered subcutaneously.

[0251] Embodiment 42. The method of embodiment 41, wherein the effective amount is 0.1 μg / kg to 5 μg / kg.

[0252] Embodiment 43. The method of embodiment 42, wherein the effective amount is 2.5 μg / kg.

[0253] Embodiment 44. The method of embodiment 41, wherein the effective amount is 6.5 μg to 750 μg.

[0254] Embodiment 45. The method of embodiment 44, wherein the effective amount is 162.5 μg to 375 μg.

[0255] Embodiment 46 The method of any one of embodiments 35 to 40, wherein the MANPs are administered intravenously.

[0256] Embodiment 47. The method of embodiment 46, wherein the effective amount is 10 pmol / kg / min to 100 nmol / kg / min.

[0257] Embodiment 48 The method of any one of embodiments 35 to 40, wherein the MANP is administered intravenously followed by subcutaneous administration.

[0258] Embodiment 49. The method of embodiment 48, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 0.1 μg / kg to 5 μg / kg.

[0259] Embodiment 50. The method of embodiment 49, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 2.5 μg / kg.

[0260] Embodiment 51. The method of embodiment 48, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 6.5 μg to 750 μg.

[0261] Embodiment 52. The method of embodiment 51, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 162.5 μg to 375 μg.

[0262] Embodiment 53. The patient is taking 25 kg / m 2 The method of any one of embodiments 35 to 52, wherein the patient has a higher body mass index (BMI).

[0263] Embodiment 54. The patient is taking 30 kg / m 2 The method of any one of embodiments 35 to 53, wherein the patient has a higher BMI.

[0264] Embodiment 55. The patient is taking 35 kg / m 2 The method of any one of embodiments 35 to 54, wherein the patient has a higher BMI.

[0265] Embodiment 56. The method of any one of embodiments 35 to 55, wherein the patient exhibits a sitting systolic blood pressure of 140 mmHg to 180 mmHg or a sitting diastolic blood pressure of 90 mmHg to 100 mmHg prior to said administering.

[0266] Embodiment 57. The method of any one of embodiments 35 to 56, wherein the patient has been taking an antihypertensive drug for 30 days prior to said administering, or has been taking a statin, ezetimibe, or a combination thereof for 60 days prior to said administering.

[0267] Embodiment 58. The method of any one of embodiments 35 to 57, wherein the patient exhibits at least two characteristics selected from the group consisting of abdominal obesity, high serum triglyceride levels, low serum high-density lipoprotein (HDL) cholesterol levels, and high fasting plasma glucose levels.

[0268] Embodiment 59. The method of embodiment 58, wherein the abdominal obesity is identified by a waist circumference of 102 cm or more in a male patient or 88 cm or more in a female patient.

[0269] Embodiment 60. The method of embodiment 58 or 59, wherein the elevated serum triglyceride level is identified by a serum triglyceride level of 150 mg / dL or greater in the patient prior to said administering.

[0270] Embodiment 61. The method of any one of embodiments 58 to 60, wherein the low HDL cholesterol level is identified by an HDL cholesterol level of less than 40 mg / dL in a male patient or an HDL cholesterol level of less than 50 mg / dL in a female patient.

[0271] Embodiment 62. The method of any one of embodiments 58 to 60, wherein the low HDL cholesterol level is identified in the patient receiving medication to increase HDL cholesterol levels for that low HDL cholesterol level.

[0272] Embodiment 63. The method of any one of embodiments 58 to 62, wherein the high fasting plasma glucose level is identified in the patient by a fasting plasma glucose level of 100 mg / dL or greater.

[0273] Embodiment 64. The method of any one of embodiments 58 to 62, wherein the high fasting plasma glucose level is identified in the patient receiving medication for elevated blood glucose levels to lower blood glucose levels.

[0274] Embodiment 65. The patient is known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; Taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration Having received an investigational drug or used an investigational device within 30 days prior to the administration of the investigational drug. a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or substance abuse or addiction (within two years prior to said administration); Having a history of difficulty in donating blood, or having donated blood or blood products within 45 days prior to the administration. The patient has had a clinically significant new illness within one month prior to the administration of the drug. A history of severe allergies coronary artery disease, cerebrovascular disease, or a history of syncope; a history of epilepsy or other seizure disorder; Having had an organ transplant; and Having had a malignant tumor within 5 years prior to the administration of the above. 65. The method of any one of embodiments 35 to 64, wherein the method does not exhibit any property selected from the group consisting of:

[0275] Embodiment 66. The patient is known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, History of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and Taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration 66. The method of any one of embodiments 35 to 65, wherein the medicament does not exhibit any property selected from the group consisting of:

[0276] Embodiment 67. The method of any one of embodiments 35 to 66, wherein the patient's plasma glucose level is reduced by 1 to 17 mg / mL relative to the baseline plasma glucose level within 4 hours after the administration.

[0277] Embodiment 68. The method of any one of embodiments 35 to 67, further comprising, consisting essentially of, or consisting of administering a second drug selected from the group consisting of a diuretic, angiotensin-converting enzyme (ACE) inhibitor, angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

[0278] Embodiment 69. A method of increasing plasma atrial natriuretic peptide (ANP)-like peptide levels, applied to a patient in need of such treatment having a baseline plasma cyclic guanosine monophosphate (cGMP) level below 10 pmol / mL, comprising, or consisting essentially of, or consisting of, administering to said patient an effective amount of alternatively spliced ​​atrial natriuretic peptide (MANP), thereby increasing plasma cGMP levels relative to said baseline.

[0279] Embodiment 70. The method of embodiment 69, wherein the baseline plasma cGMP level is less than 8 pmol / mL.

[0280] Embodiment 71. The method of embodiment 69 or 70, wherein increasing the plasma cGMP level is increasing the plasma cGMP level from the baseline by 2 to 8 pmol / mL within 0.5 hours after the administration.

[0281] Embodiment 72. The method of embodiment 69 or 70, wherein increasing the plasma cGMP level is increasing the plasma cGMP level from the baseline by 1 to 5 pmol / mL within 0.5 to 4 hours after the administration.

[0282] Embodiment 73. The method of any one of embodiments 69 to 72, wherein increasing the ANP-like peptide level is increasing the ANP-like peptide level by 1 to 70 pg / mL from a baseline ANP-like peptide level within 0.5 hours after the administration.

[0283] Embodiment 74. The method of any one of embodiments 69 to 72, wherein increasing the ANP-like peptide level is increasing the ANP-like peptide level by 1 to 40 pg / mL from a baseline ANP-like peptide level within 1 hour after the administration.

[0284] Embodiment 75. The method of any one of embodiments 69 to 74, wherein the MANPs are administered subcutaneously.

[0285] Embodiment 76. The method of embodiment 75, wherein the effective amount is 0.1 μg / kg to 5 μg / kg.

[0286] Embodiment 77. The method of embodiment 76, wherein the effective amount is 2.5 μg / kg.

[0287] Embodiment 78. The method of embodiment 75, wherein the effective amount is 6.5 μg to 750 μg.

[0288] Embodiment 79. The method of embodiment 78, wherein the effective amount is 162.5 μg to 375 μg.

[0289] Embodiment 80. The method of any one of embodiments 69 to 74, wherein the MANPs are administered intravenously.

[0290] Embodiment 81. The method of embodiment 80, wherein the effective amount is 10 pmol / kg / min to 100 nmol / kg / min.

[0291] Embodiment 82 The method of any one of embodiments 69 to 74, wherein the MANP is administered intravenously followed by subcutaneous administration.

[0292] Embodiment 83. The method of embodiment 82, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 0.1 μg / kg to 5 μg / kg.

[0293] Embodiment 84. The method of embodiment 83, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 2.5 μg / kg.

[0294] Embodiment 85. The method of embodiment 82, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 6.5 μg to 750 μg.

[0295] Embodiment 86. The method of embodiment 85, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 162.5 μg to 375 μg.

[0296] Embodiment 87. The patient is taking 25 kg / m 2 The method of any one of embodiments 69 to 86, wherein the patient has a higher body mass index (BMI).

[0297] Embodiment 88. The patient is taking 30 kg / m 2 The method of any one of embodiments 69 to 87, wherein the patient has a higher BMI.

[0298] Embodiment 89. The patient is taking 35 kg / m 2 The method of any one of embodiments 69 to 88, wherein the patient has a higher BMI.

[0299] Embodiment 90. The method of any one of embodiments 69 to 89, wherein the patient exhibits a sitting systolic blood pressure of 140 mmHg to 180 mmHg or a sitting diastolic blood pressure of 90 mmHg to 100 mmHg prior to said administering.

[0300] Embodiment 91. The method of any one of embodiments 69 to 90, wherein the patient has been taking an antihypertensive drug for 30 days prior to said administering, or has been taking a statin, ezetimibe, or a combination thereof for 60 days prior to said administering.

[0301] Embodiment 92. The method of any one of embodiments 69 to 91, wherein the patient exhibits at least two characteristics selected from the group consisting of abdominal obesity, high serum triglyceride levels, low serum high-density lipoprotein (HDL) cholesterol levels, and high fasting plasma glucose levels.

[0302] Embodiment 93. The method of embodiment 92, wherein the abdominal obesity is identified by a waist circumference of 102 cm or more in a male patient or 88 cm or more in a female patient.

[0303] Embodiment 94. The method of embodiment 92 or 93, wherein the elevated serum triglyceride level is identified by a serum triglyceride level of 150 mg / dL or greater in the patient prior to the administering.

[0304] Embodiment 95. The method of any one of embodiments 92 to 94, wherein the low HDL cholesterol level is identified by an HDL cholesterol level of less than 40 mg / dL in a male patient or an HDL cholesterol level of less than 50 mg / dL in a female patient.

[0305] Embodiment 96. The method of any one of embodiments 92 to 94, wherein the low HDL cholesterol level is identified in the patient receiving medication to increase HDL cholesterol levels for the low HDL cholesterol level.

[0306] Embodiment 97. The method of any one of embodiments 92 to 96, wherein the high fasting plasma glucose level is identified in the patient by a fasting plasma glucose level of 100 mg / dL or greater.

[0307] Embodiment 98. The method of any one of embodiments 92 to 96, wherein the high fasting plasma glucose level is identified in the patient receiving medication for elevated blood glucose levels to lower blood glucose levels.

[0308] Embodiment 99. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; Taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration Having received an investigational drug or used an investigational device within 30 days prior to the administration of the investigational drug. a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or substance abuse or addiction (within two years prior to said administration); Having a history of difficulty in donating blood, or having donated blood or blood products within 45 days prior to the administration. The patient has had a clinically significant new illness within one month prior to the administration of the drug. A history of severe allergies coronary artery disease, cerebrovascular disease, or a history of syncope; a history of epilepsy or other seizure disorder; Having had an organ transplant; and Having had a malignant tumor within 5 years prior to the administration of the above. 99. The method of any one of embodiments 69 to 98, wherein the method does not exhibit any feature selected from the group consisting of:

[0309] Embodiment 100. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, History of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and Taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration 99. The method of any one of embodiments 69 to 99, wherein the method does not exhibit any property selected from the group consisting of:

[0310] Embodiment 101. The method of any one of embodiments 69 to 100, wherein the patient's sitting systolic blood pressure is reduced by 0.1 to 15% relative to baseline sitting systolic blood pressure within 12 hours after said administration.

[0311] Embodiment 102. The method of any one of embodiments 69 to 101, wherein the patient's sitting diastolic blood pressure is reduced by 0.1 to 5% relative to baseline sitting diastolic blood pressure within 6 hours after the administration.

[0312] Embodiment 103. The method of any one of embodiments 69 to 102, wherein the patient's plasma glucose level is reduced by 1 to 17 mg / mL relative to the baseline plasma glucose level within 4 hours after the administration.

[0313] Embodiment 104. The method of any one of embodiments 69 to 103, 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).

[0314] Embodiment 105. A method for lowering plasma glucose levels, applied to a patient in need of such treatment, comprising, consisting essentially of, or consisting of administering to said patient an effective amount of alternatively spliced ​​atrial natriuretic peptide (MANP).

[0315] Embodiment 106. The method of embodiment 105, wherein said reducing is a reduction in the plasma glucose level by 1 to 17 mg / mL relative to baseline plasma glucose levels within 4 hours after said administering.

[0316] Embodiment 107. The method of embodiment 105 or 106, wherein the MANPs are administered subcutaneously.

[0317] Embodiment 108. The method of embodiment 107, wherein the effective amount is 0.1 μg / kg to 5 μg / kg.

[0318] Embodiment 109. The method of embodiment 108, wherein the effective amount is 2.5 μg / kg.

[0319] Embodiment 110. The method of embodiment 107, wherein the effective amount is 6.5 μg to 750 μg.

[0320] Embodiment 111. The method of embodiment 110, wherein the effective amount is 162.5 μg to 375 μg.

[0321] Embodiment 112. The method of embodiment 105 or 106, wherein the MANPs are administered intravenously.

[0322] Embodiment 113. The method of embodiment 112, wherein the effective amount is 10 pmol / kg / min to 100 nmol / kg / min.

[0323] Embodiment 114. The method of embodiment 105 or 106, wherein the MANP is administered intravenously followed by subcutaneous administration.

[0324] Embodiment 115. The method of embodiment 114, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 0.1 μg / kg to 5 μg / kg.

[0325] Embodiment 116. The method of embodiment 115, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 2.5 μg / kg.

[0326] Embodiment 117. The method of embodiment 114, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 6.5 μg to 750 μg.

[0327] Embodiment 118. The method of embodiment 117, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 162.5 μg to 375 μg.

[0328] Embodiment 119. The patient is taking 25 kg / m 2 The method of any one of embodiments 105 to 118, wherein the patient has a higher body mass index (BMI).

[0329] Embodiment 120. The patient is taking 30 kg / m 2 120. The method of any one of embodiments 105 to 119, wherein the patient has a higher BMI.

[0330] Embodiment 121. The patient is taking 35 kg / m 2 The method of any one of embodiments 105 to 120, wherein the patient has a higher BMI.

[0331] Embodiment 122. The method of any one of embodiments 105 to 121, wherein the patient exhibits a sitting systolic blood pressure of 140 mmHg to 180 mmHg or a sitting diastolic blood pressure of 90 mmHg to 100 mmHg prior to said administration.

[0332] Embodiment 123. The method of any one of embodiments 105 to 122, wherein the patient has been taking an antihypertensive drug for 30 days prior to said administering, or has been taking a statin, ezetimibe, or a combination thereof for 60 days prior to said administering.

[0333] Embodiment 124. The method of any one of embodiments 105 to 123, wherein the patient exhibits at least two characteristics selected from the group consisting of abdominal obesity, high serum triglyceride levels, low serum high-density lipoprotein (HDL) cholesterol levels, and high fasting plasma glucose levels.

[0334] Embodiment 125. The method of embodiment 124, wherein the abdominal obesity is identified by a waist circumference of 102 cm or more in a male patient or 88 cm or more in a female patient.

[0335] Embodiment 126. The method of embodiment 124 or 125, wherein the elevated serum triglyceride level is identified by a serum triglyceride level of 150 mg / dL or greater in the patient prior to the administering.

[0336] Embodiment 127. The method of any one of embodiments 124 to 126, wherein the low HDL cholesterol level is identified by an HDL cholesterol level of less than 40 mg / dL in a male patient or an HDL cholesterol level of less than 50 mg / dL in a female patient.

[0337] Embodiment 128. The method of any one of embodiments 124 to 126, wherein the low HDL cholesterol level is identified in the patient receiving medication to increase HDL cholesterol levels for the low HDL cholesterol level.

[0338] Embodiment 129. The method of any one of embodiments 124 to 128, wherein the high fasting plasma glucose level is identified in the patient by a fasting plasma glucose level of 100 mg / dL or greater.

[0339] Embodiment 130. The method of any one of embodiments 124 to 128, wherein the high fasting plasma glucose level is identified in the patient receiving medication for elevated blood glucose levels to lower blood glucose levels.

[0340] Embodiment 131. The patient is Known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; Taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration Having received an investigational drug or used an investigational device within 30 days prior to the administration of the investigational drug. a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or substance abuse or addiction (within two years prior to said administration); Having a history of difficulty in donating blood, or having donated blood or blood products within 45 days prior to the administration. The patient has had a clinically significant new illness within one month prior to the administration of the drug. A history of severe allergies coronary artery disease, cerebrovascular disease, or a history of syncope; a history of epilepsy or other seizure disorder; Having had an organ transplant; and Having had a malignant tumor within 5 years prior to the administration of the above. 131. The method of any one of embodiments 105 to 130, wherein the method does not exhibit any feature selected from the group consisting of:

[0341] Embodiment 132. The patient is known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, History of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and Taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration 132. The method of any one of embodiments 105 to 131, wherein the method does not exhibit any property selected from the group consisting of:

[0342] Embodiment 133. The method of any one of embodiments 105 to 131, wherein the patient's sitting systolic blood pressure is reduced by 0.1 to 15% relative to baseline sitting systolic blood pressure within 12 hours after said administration.

[0343] Embodiment 134. The method of any one of embodiments 105 to 132, wherein the patient's sitting diastolic blood pressure is reduced by 0.1 to 5% relative to baseline sitting diastolic blood pressure within 6 hours after the administration.

[0344] Embodiment 135. The method of any one of embodiments 105 to 133, further comprising, consisting essentially of, or consisting of administering a second drug selected from the group consisting of a diuretic, angiotensin-converting enzyme (ACE) inhibitor, angiotensin II receptor blocker (ARB), and a calcium channel blocker (CCB).

[0345] Embodiment 136. A method for improving insulin sensitivity, applied to a patient in need thereof, comprising, consisting essentially of, or consisting of administering to said patient an effective amount of alternatively spliced ​​atrial natriuretic peptide (MANP).

[0346] Embodiment 137. The method of embodiment 136, wherein the improvement in insulin sensitivity is an increase in the patient's updated homeostasis model assessment insulin sensitivity (HOMA2-S) value by 0.1 to 35% relative to the patient's baseline HOMA2-S value within 4 hours after the administration.

[0347] Embodiment 138. The method of embodiment 136, wherein the improvement in insulin sensitivity is a decrease of 0.1 to 0.6 in the patient's updated homeostasis model assessment of insulin resistance (HOMA2-IR) value from the baseline HOMA2-IR value within 4 hours after the administration.

[0348] Embodiment 139. The method of any one of embodiments 136 to 138, wherein the MANPs are administered subcutaneously.

[0349] Embodiment 140. The method of embodiment 139, wherein the effective amount is 0.1 μg / kg to 5 μg / kg.

[0350] Embodiment 141. The method of embodiment 140, wherein the effective amount is 2.5 μg / kg.

[0351] Embodiment 142. The method of embodiment 139, wherein the effective amount is 6.5 μg to 750 μg.

[0352] Embodiment 143. The method of embodiment 142, wherein the effective amount is 162.5 μg to 375 μg.

[0353] Embodiment 144. The method of any one of embodiments 136 to 138, wherein the MANP is administered intravenously.

[0354] Embodiment 145. The method of embodiment 144, wherein the effective amount is 10 pmol / kg / min to 100 nmol / kg / min.

[0355] Embodiment 146. The method of any one of embodiments 136 to 138, wherein the MANP is administered intravenously followed by subcutaneous administration.

[0356] Embodiment 147. The method of embodiment 146, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 0.1 μg / kg to 5 μg / kg.

[0357] Embodiment 148. The method of embodiment 147, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 2.5 μg / kg.

[0358] Embodiment 149. The method of embodiment 146, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 6.5 μg to 750 μg.

[0359] Embodiment 150. The method of embodiment 149, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 162.5 μg to 375 μg.

[0360] Embodiment 151. The patient is taking 25 kg / m 2 The method of any one of embodiments 136 to 150, wherein the patient has a higher body mass index (BMI).

[0361] Embodiment 152. The patient is taking 30 kg / m 2 The method of any one of embodiments 136 to 151, wherein the patient has a higher BMI.

[0362] Embodiment 153. The patient is taking 35 kg / m 2 The method of any one of embodiments 136 to 152, wherein the patient has a higher BMI.

[0363] Embodiment 154. The method of any one of embodiments 136 to 153, wherein the patient exhibits a sitting systolic blood pressure of 140 mmHg to 180 mmHg or a sitting diastolic blood pressure of 90 mmHg to 100 mmHg prior to said administration.

[0364] Embodiment 155. The method of any one of embodiments 136 to 154, wherein the patient has been taking an antihypertensive drug for 30 days prior to said administering, or has been taking a statin, ezetimibe, or a combination thereof for 60 days prior to said administering.

[0365] Embodiment 156. The method of any one of embodiments 136 to 155, wherein the patient exhibits at least two characteristics selected from the group consisting of abdominal obesity, high serum triglyceride levels, low serum high-density lipoprotein (HDL) cholesterol levels, and high fasting plasma glucose levels.

[0366] Embodiment 157. The method of embodiment 156, wherein the abdominal obesity is identified by a waist circumference of 102 cm or more in a male patient or 88 cm or more in a female patient.

[0367] Embodiment 158. The method of embodiment 156 or 157, wherein the elevated serum triglyceride level is identified by a serum triglyceride level of 150 mg / dL or greater in the patient prior to said administering.

[0368] Embodiment 159. The method of any one of embodiments 156 to 158, wherein the low HDL cholesterol level is identified by an HDL cholesterol level of less than 40 mg / dL in a male patient or an HDL cholesterol level of less than 50 mg / dL in a female patient.

[0369] Embodiment 160. The method of any one of embodiments 156 to 158, wherein the low HDL cholesterol level is identified in the patient receiving medication for low HDL cholesterol levels to increase HDL cholesterol levels.

[0370] Embodiment 161. The method of any one of embodiments 156 to 160, wherein the high fasting plasma glucose level is identified in the patient by a fasting plasma glucose level of 100 mg / dL or greater.

[0371] Embodiment 162. The method of any one of embodiments 156 to 160, wherein the high fasting plasma glucose level is identified in the patient receiving medication for elevated blood glucose levels to lower blood glucose levels.

[0372] Embodiment 163. The patient is known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; Taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration Having received an investigational drug or used an investigational device within 30 days prior to the administration of the investigational drug. a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or substance abuse or addiction (within two years prior to said administration); Having a history of difficulty in donating blood, or having donated blood or blood products within 45 days prior to the administration. The patient has had a clinically significant new illness within one month prior to the administration of the drug. A history of severe allergies coronary artery disease, cerebrovascular disease, or a history of syncope; a history of epilepsy or other seizure disorder; Having had an organ transplant; and Having had a malignant tumor within 5 years prior to the administration of the above. 163. The method of any one of embodiments 136 to 162, wherein the method does not exhibit any property selected from the group consisting of:

[0373] Embodiment 164. The patient is known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, History of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and Taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration 164. The method of any one of embodiments 136 to 163, wherein the method does not exhibit any property selected from the group consisting of:

[0374] Embodiment 165. The method of any one of embodiments 136 to 164, wherein the patient's sitting systolic blood pressure is reduced by 0.1 to 15% relative to baseline sitting systolic blood pressure within 12 hours after said administration.

[0375] Embodiment 166. The method of any one of embodiments 136 to 165, wherein the patient's sitting diastolic blood pressure is reduced by 0.1 to 5% relative to baseline sitting diastolic blood pressure within 6 hours after the administration.

[0376] Embodiment 167. The method of any one of embodiments 136 to 166, wherein the patient's plasma glucose level is reduced by 1 to 17 mg / mL relative to the baseline plasma glucose level within 4 hours after the administration.

[0377] Embodiment 168. The method of any one of embodiments 136 to 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).

[0378] Embodiment 169. A method for increasing plasma non-esterified fatty acid (NEFA) levels, applied to a patient in need of such treatment, comprising, consisting essentially of, or consisting of administering to said patient an effective amount of alternatively spliced ​​atrial natriuretic peptide (MANP).

[0379] Embodiment 170. The method of embodiment 169, wherein said increasing is an increase in the plasma NEFA level by 1 μM from the baseline NEFA level within 1 hour after said administration.

[0380] Embodiment 171. The method of embodiment 169 or 170, wherein the MANPs are administered subcutaneously.

[0381] Embodiment 172. The method of embodiment 171, wherein the effective amount is 0.1 μg / kg to 5 μg / kg.

[0382] Embodiment 173. The method of embodiment 172, wherein the effective amount is 2.5 μg / kg.

[0383] Embodiment 174. The method of embodiment 171, wherein the effective amount is 6.5 μg to 750 μg.

[0384] Embodiment 175. The method of embodiment 174, wherein the effective amount is 162.5 μg to 375 μg.

[0385] Embodiment 176. The method of embodiment 169 or 170, wherein the MANP is administered intravenously.

[0386] Embodiment 177. The method of embodiment 176, wherein the effective amount is 10 pmol / kg / min to 100 nmol / kg / min.

[0387] Embodiment 178. The method of embodiment 169 or 170, wherein the MANP is administered intravenously followed by subcutaneous administration.

[0388] Embodiment 179. The method of embodiment 178, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 0.1 μg / kg to 5 μg / kg.

[0389] Embodiment 180. The method of embodiment 179, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 2.5 μg / kg.

[0390] Embodiment 181. The method of embodiment 178, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 6.5 μg to 750 μg.

[0391] Embodiment 182. The method of embodiment 181, wherein the MANP is administered intravenously at a dose of 10 pmol / kg / min to 100 nmol / kg / min, followed by subcutaneous administration at a dose of 162.5 μg to 375 μg.

[0392] Embodiment 183. The patient is taking 25 kg / m 2 The method of any one of embodiments 169 to 182, wherein the patient has a higher body mass index (BMI).

[0393] Embodiment 184. The patient is taking a 30 kg / m 2 The method of any one of embodiments 169 to 183, wherein the patient has a higher BMI.

[0394] Embodiment 185. The patient is taking 35 kg / m 2 The method of any one of embodiments 169 to 184, wherein the patient has a higher BMI.

[0395] Embodiment 186. The method of any one of embodiments 169 to 185, wherein the patient exhibits a sitting systolic blood pressure of 140 mmHg to 180 mmHg or a sitting diastolic blood pressure of 90 mmHg to 100 mmHg prior to said administration.

[0396] Embodiment 187. The method of any one of embodiments 169 to 186, wherein the patient has been taking an antihypertensive drug for 30 days prior to said administering, or has been taking a statin, ezetimibe, or a combination thereof for 60 days prior to said administering.

[0397] Embodiment 188. The method of any one of embodiments 169 to 187, wherein the patient exhibits at least two characteristics selected from the group consisting of abdominal obesity, high serum triglyceride levels, low serum high-density lipoprotein (HDL) cholesterol levels, and high fasting plasma glucose levels.

[0398] Embodiment 189. The method of embodiment 188, wherein the abdominal obesity is identified by a waist circumference of 102 cm or more in male patients and 88 cm or more in female patients.

[0399] Embodiment 190. The method of embodiment 188 or 189, wherein the elevated serum triglyceride level is identified by a serum triglyceride level of 150 mg / dL or greater in the patient prior to said administering.

[0400] Embodiment 191. The method of any one of embodiments 188 to 190, wherein the low HDL cholesterol level is identified by an HDL cholesterol level of less than 40 mg / dL in a male patient or an HDL cholesterol level of less than 50 mg / dL in a female patient.

[0401] Embodiment 192. The method of any one of embodiments 188 to 190, wherein the low HDL cholesterol level is identified in the patient receiving drug treatment for low HDL cholesterol levels to increase HDL cholesterol levels.

[0402] Embodiment 193. The method of any one of embodiments 188 to 192, wherein the high fasting plasma glucose level is identified in the patient by a fasting plasma glucose level of 100 mg / dL or greater.

[0403] Embodiment 194. The method of any one of embodiments 188 to 192, wherein the high fasting plasma glucose level is identified in the patient receiving medication for elevated blood glucose levels to lower blood glucose levels.

[0404] Embodiment 195. The patient is known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; Taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration Having received an investigational drug or used an investigational device within 30 days prior to the administration of the investigational drug. a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or substance abuse or addiction (within two years prior to said administration); Having a history of difficulty in donating blood, or having donated blood or blood products within 45 days prior to the administration. The patient has had a clinically significant new illness within one month prior to the administration of the drug. A history of severe allergies coronary artery disease, cerebrovascular disease, or a history of syncope; a history of epilepsy or other seizure disorder; Having had an organ transplant; and Having had a malignant tumor within 5 years prior to the administration of the above. 195. The method of any one of embodiments 169 to 194, wherein the method does not exhibit any property selected from the group consisting of:

[0405] Embodiment 196. The patient is known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, History of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and Taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to the administration 196. The method of any one of embodiments 169 to 195, wherein the method does not exhibit any property selected from the group consisting of:

[0406] Embodiment 197. The method of any one of embodiments 169 to 196, wherein the patient's sitting systolic blood pressure is reduced by 0.1 to 15% relative to baseline sitting systolic blood pressure within 12 hours after said administration.

[0407] Embodiment 198. The method of any one of embodiments 169 to 197, wherein the patient's sitting diastolic blood pressure is reduced by 0.1 to 5% relative to baseline sitting diastolic blood pressure within 6 hours after the administration.

[0408] Embodiment 199. The method of any one of embodiments 169 to 198, wherein the patient's plasma glucose level is reduced by 1 to 17 mg / mL relative to the baseline plasma glucose level within 4 hours after the administration.

[0409] Embodiment 200. The method of any one of embodiments 169 to 199, 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).

Claims

1. 1. A method for treating hypertension with metabolic syndrome, administered to a patient in need of such treatment who has a baseline plasma cyclic guanosine monophosphate (cGMP) level below 10 pmol / mL, comprising administering to the patient an effective amount of alternatively spliced ​​atrial natriuretic peptide (MANP) to increase the plasma cGMP level compared to baseline.

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

3. 3. The method of claim 1, wherein said increasing is an increase of 2-8 pmol / mL from the baseline plasma cGMP level within 0.5 hours after said administration.

4. 3. The method of claim 1, wherein said increasing is an increase of 1 to 5 pmol / mL from the baseline plasma cGMP level within 0.5 to 4 hours after said administration.

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

6. 6. The method of claim 5, wherein the effective amount is 0.1 μg / kg to 5 μg / kg.

7. 7. The method of claim 6, wherein the effective amount is 2.5 μg / kg.

8. 6. The method of claim 5, wherein the effective amount is between 6.5 μg and 750 μg.

9. 9. The method of claim 8, wherein the effective amount is between 162.5 μg and 375 μg.

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

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

12. 5. The method of any one of claims 1 to 4, wherein the MANP is administered intravenously followed by subcutaneous administration.

13. 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, followed by subcutaneous administration at a dose of 0.1 μg / kg to 5 μg / kg.

14. 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, followed by subcutaneous administration at a dose of 2.5 μg / kg.

15. 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, followed by subcutaneous administration at a dose of 6.5 μg to 750 μg.

16. 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, followed by subcutaneous administration at a dose of 162.5 μg to 375 μg.

17. The patient is 25 kg / m ２ 17. The method of any one of claims 1 to 16, wherein the patient has a higher body mass index (BMI).

18. The patient is 30 kg / m ２ 18. The method of any one of claims 1 to 17, wherein the patient has a higher BMI.

19. The patient is 35 kg / m ２ 19. The method of any one of claims 1 to 18, wherein the patient has a higher BMI.

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

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

22. 22. The method of claim 20 or 21, wherein the elevated serum triglyceride level is identified by a serum triglyceride level of 150 mg / dL or greater in the patient prior to the administration.

23. 23. The method of any one of claims 20-22, wherein the low HDL cholesterol level is identified by an HDL cholesterol level of less than 40 mg / dL in a male patient or an HDL cholesterol level of less than 50 mg / dL in a female patient.

24. 23. The method of any one of claims 20 to 22, wherein the low HDL cholesterol level is identified in the patient receiving medication for low HDL cholesterol to increase the HDL cholesterol level.

25. 25. The method of any one of claims 20-24, wherein the elevated fasting plasma glucose level is identified in the patient by a fasting plasma glucose level of 100 mg / dL or greater.

26. 25. The method of any one of claims 20 to 24, wherein the high fasting plasma glucose level is identified in the patient receiving medication for elevated blood glucose levels to lower blood glucose levels.

27. 27. The method of any one of claims 1 to 26, wherein the patient exhibits a sitting systolic blood pressure of 140 mmHg to 180 mmHg or a sitting diastolic blood pressure of 90 mmHg to 100 mmHg prior to said administering.

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

29. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to said administration; The patient has received an investigational drug or used an investigational device within 30 days prior to the administration. a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or drug abuse or addiction (within the two years prior to said administration); Having a history of difficulty in donating blood, or having donated blood or blood products within 45 days prior to the administration. The patient has had a clinically significant new illness within one month prior to the administration of the drug. A history of severe allergies coronary artery disease, cerebrovascular disease, or a history of syncope; a history of epilepsy or other seizure disorder; Having had an organ transplant; and Having a malignant tumor within 5 years prior to the administration of the above-mentioned drug.

29. The method of any one of claims 1 to 28, wherein the method does not exhibit any property selected from the group consisting of:

30. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, History of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to said administration; 30. The method of any one of claims 1 to 29, wherein the method does not exhibit any property selected from the group consisting of:

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

32. 32. The method of any one of claims 1-31, wherein the patient's sitting diastolic blood pressure is reduced by 0.1 to 5% relative to baseline sitting diastolic blood pressure within 6 hours after said administration.

33. 33. The method of any one of claims 1-32, wherein the patient's plasma glucose level is reduced by 1-17 mg / mL relative to the baseline plasma glucose level within 4 hours after the administration.

34. 34. The method of 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. 1. A method for lowering blood pressure in a patient in need of such treatment who has a baseline plasma cyclic guanosine monophosphate (cGMP) level below 10 pmol / mL, comprising administering to the patient an effective amount of alternatively spliced ​​atrial natriuretic peptide (MANP), thereby increasing the plasma cGMP level compared to the baseline.

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

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

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

39. 39. The method of any one of claims 35-38, wherein said reducing is a 0.1 to 15% decrease in the patient's sitting systolic blood pressure relative to baseline sitting systolic blood pressure within 12 hours after said administration.

40. 39. The method of any one of claims 35-38, wherein said reducing is a 0.1 to 5% decrease in the patient's sitting diastolic blood pressure within 6 hours after said administration, relative to baseline sitting diastolic blood pressure.

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

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

43. 43. The method of claim 42, wherein the effective amount is 2.5 μg / kg.

44. 42. The method of claim 41, wherein the effective amount is between 6.5 μg and 750 μg.

45. 45. The method of claim 44, wherein the effective amount is between 162.5 μg and 375 μg.

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

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

48. 41. The method of any one of claims 35 to 40, wherein the MANP is administered intravenously followed by subcutaneous administration.

49. 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, followed by subcutaneous administration at a dose of 0.1 μg / kg to 5 μg / kg.

50. 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, followed by subcutaneous administration at a dose of 2.5 μg / kg.

51. 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, followed by subcutaneous administration at a dose of 6.5 μg to 750 μg.

52. 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, followed by subcutaneous administration at a dose of 162.5 μg to 375 μg.

53. The patient is 25 kg / m ２ 53. The method of any one of claims 35 to 52, wherein the patient has a higher body mass index (BMI).

54. The patient is 30 kg / m ２ 54. The method of any one of claims 35 to 53, wherein the patient has a higher BMI.

55. The patient is 35 kg / m ２ 55. The method of any one of claims 35 to 54, wherein the patient has a higher BMI.

56. 56. The method of any one of claims 35-55, wherein the patient exhibits a sitting systolic blood pressure of 140 mmHg to 180 mmHg or a sitting diastolic blood pressure of 90 mmHg to 100 mmHg prior to said administering.

57. 57. The method of any one of claims 35-56, wherein the patient has been taking an antihypertensive drug for 30 days prior to said administering, or has been taking a statin, ezetimibe, or a combination thereof for 60 days prior to said administering.

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

59. 59. The method of claim 58, wherein said abdominal obesity is identified by a waist circumference of 102 cm or greater in said patient who is male and 88 cm or greater in said patient who is female.

60. 60. The method of claim 58 or 59, wherein the elevated serum triglyceride level is identified by a serum triglyceride level of 150 mg / dL or greater in the patient prior to the administration.

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

62. 61. The method of any one of claims 58 to 60, wherein the low HDL cholesterol level is identified in the patient receiving medication for low HDL cholesterol to increase HDL cholesterol levels.

63. 63. The method of any one of claims 58-62, wherein the high fasting plasma glucose level is identified in the patient by a fasting plasma glucose level of 100 mg / dL or greater.

64. 63. The method of any one of claims 58 to 62, wherein the high fasting plasma glucose level is identified in the patient receiving medication for elevated blood glucose levels to lower blood glucose levels.

65. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to said administration; The patient has received an investigational drug or used an investigational device within 30 days prior to the administration. a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or drug abuse or addiction (within the two years prior to said administration); Having a history of difficulty in donating blood, or having donated blood or blood products within 45 days prior to the administration. The patient has had a clinically significant new illness within one month prior to the administration of the drug. A history of severe allergies coronary artery disease, cerebrovascular disease, or a history of syncope; a history of epilepsy or other seizure disorder; Having had an organ transplant; and Having a malignant tumor within 5 years prior to the administration of the above-mentioned drug.

65. The method of any one of claims 35 to 64, wherein the compound does not exhibit any property selected from the group consisting of:

66. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, History of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to said administration; 66. The method of any one of claims 35 to 65, wherein the compound does not exhibit any property selected from the group consisting of:

67. 67. The method of any one of claims 35-66, wherein the patient's plasma glucose level is reduced by 1-17 mg / mL relative to the baseline plasma glucose level within 4 hours after the administration.

68. 68. The method of 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. 1. A method for increasing plasma atrial natriuretic peptide (ANP)-like peptide levels, the method being applied to a patient in need of such treatment who has a baseline plasma cyclic guanosine monophosphate (cGMP) level below 10 pmol / mL, comprising administering to the patient an effective amount of alternatively spliced ​​atrial natriuretic peptide (MANP), thereby increasing the plasma cGMP level compared to the baseline.

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

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

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

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

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

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

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

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

78. 76. The method of claim 75, wherein the effective amount is between 6.5 μg and 750 μg.

79. 79. The method of claim 78, wherein the effective amount is between 162.5 μg and 375 μg.

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

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

82. 75. The method of any one of claims 69 to 74, wherein the MANP is administered intravenously followed by subcutaneous administration.

83. 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, followed by subcutaneous administration at a dose of 0.1 μg / kg to 5 μg / kg.

84. 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, followed by subcutaneous administration at a dose of 2.5 μg / kg.

85. 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, followed by subcutaneous administration at a dose of 6.5 μg to 750 μg.

86. 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, followed by subcutaneous administration at a dose of 162.5 μg to 375 μg.

87. The patient is 25 kg / m ２ 87. The method of any one of claims 69 to 86, wherein the patient has a higher body mass index (BMI).

88. The patient is 30 kg / m ２ 88. The method of any one of claims 69 to 87, wherein the patient has a higher BMI.

89. The patient is 35 kg / m ２ 89. The method of any one of claims 69 to 88, wherein the patient has a higher BMI.

90. 90. The method of any one of claims 69-89, wherein the patient exhibits a sitting systolic blood pressure of 140 mmHg to 180 mmHg or a sitting diastolic blood pressure of 90 mmHg to 100 mmHg prior to said administering.

91. 91. The method of any one of claims 69-90, wherein the patient has been taking an antihypertensive drug for 30 days prior to said administering, or has been taking a statin, ezetimibe, or a combination thereof for 60 days prior to said administering.

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

93. 93. The method of claim 92, wherein said abdominal obesity is identified by a waist circumference of 102 cm or greater in said patient who is male and 88 cm or greater in said patient who is female.

94. 94. The method of claim 92 or 93, wherein the elevated serum triglyceride level is identified by a serum triglyceride level of 150 mg / dL or greater in the patient prior to the administration.

95. 95. The method of any one of claims 92-94, wherein the low HDL cholesterol level is identified by an HDL cholesterol level of less than 40 mg / dL in a male patient or an HDL cholesterol level of less than 50 mg / dL in a female patient.

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

97. 97. The method of any one of claims 92-96, wherein the high fasting plasma glucose level is identified in the patient by a fasting plasma glucose level of 100 mg / dL or greater.

98. 97. The method of any one of claims 92 to 96, wherein the high fasting plasma glucose level is identified in the patient receiving medication for elevated blood glucose levels to lower blood glucose levels.

99. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to said administration; The patient has received an investigational drug or used an investigational device within 30 days prior to the administration. a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or drug abuse or addiction (within the two years prior to said administration); Having a history of difficulty in donating blood, or having donated blood or blood products within 45 days prior to the administration. The patient has had a clinically significant new illness within one month prior to the administration of the drug. A history of severe allergies coronary artery disease, cerebrovascular disease, or a history of syncope; a history of epilepsy or other seizure disorder; Having had an organ transplant; and Having a malignant tumor within 5 years prior to the administration of the above-mentioned drug.

99. The method of any one of claims 69 to 98, wherein the compound does not exhibit any property selected from the group consisting of:

100. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, History of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to said administration; 100. The method of any one of claims 69 to 99, wherein the compound does not exhibit any property selected from the group consisting of:

101. 101. The method of any one of claims 69-100, wherein the patient's sitting systolic blood pressure is reduced by 0.1 to 15% relative to baseline sitting systolic blood pressure within 12 hours after said administration.

102. 102. The method of any one of claims 69-101, wherein the patient's sitting diastolic blood pressure is reduced by 0.1 to 5% relative to baseline sitting diastolic blood pressure within 6 hours after said administration.

103. 103. The method of any one of claims 69-102, wherein the patient's plasma glucose level is reduced by 1-17 mg / mL relative to the baseline plasma glucose level within 4 hours after said administration.

104. 104. The method of 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 lowering plasma glucose levels, administered to a patient in need of such treatment, comprising administering to said patient an effective amount of alternatively spliced ​​atrial natriuretic peptide (MANP).

106. 106. The method of claim 105, wherein said reducing reduces said plasma glucose level by 1 to 17 mg / mL relative to baseline plasma glucose levels within 4 hours after said administration.

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

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

109. 109. The method of claim 108, wherein the effective amount is 2.5 μg / kg.

110. 108. The method of claim 107, wherein the effective amount is between 6.5 μg and 750 μg.

111. 111. The method of claim 110, wherein the effective amount is between 162.5 μg and 375 μg.

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

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

114. 107. The method of claim 105 or 106, wherein the MANP is administered intravenously followed by subcutaneous administration.

115. 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, followed by subcutaneous administration at a dose of 0.1 μg / kg to 5 μg / kg.

116. 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, followed by subcutaneous administration at a dose of 2.5 μg / kg.

117. 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, followed by subcutaneous administration at a dose of 6.5 μg to 750 μg.

118. 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, followed by subcutaneous administration at a dose of 162.5 μg to 375 μg.

119. The patient is 25 kg / m ２ 119. The method of any one of claims 105 to 118, wherein the patient has a higher body mass index (BMI).

120. The patient is 30 kg / m ２ 120. The method of any one of claims 105 to 119, wherein the patient has a higher BMI.

121. The patient is 35 kg / m ２ 121. The method of any one of claims 105 to 120, wherein the patient has a higher BMI.

122. 122. The method of any one of claims 105-121, wherein the patient exhibits a sitting systolic blood pressure of 140 mmHg to 180 mmHg or a sitting diastolic blood pressure of 90 mmHg to 100 mmHg prior to said administering.

123. 123. The method of any one of claims 105-122, wherein the patient has been taking an antihypertensive drug for 30 days prior to said administering, or has been taking a statin, ezetimibe, or a combination thereof for 60 days prior to said administering.

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

125. 125. The method of claim 124, wherein the abdominal obesity is identified by a waist circumference of 102 cm or greater in the patient who is male and 88 cm or greater in the patient who is female.

126. 126. The method of claim 124 or 125, wherein the elevated serum triglyceride level is identified by a serum triglyceride level of 150 mg / dL or greater in the patient prior to the administration.

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

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

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

130. 129. The method of any one of claims 124 to 128, wherein the high fasting plasma glucose level is identified in the patient receiving medication for elevated blood glucose levels to lower blood glucose levels.

131. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to said administration; The patient has received an investigational drug or used an investigational device within 30 days prior to the administration. a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or drug abuse or addiction (within the two years prior to said administration); Having a history of difficulty in donating blood, or having donated blood or blood products within 45 days prior to the administration. The patient has had a clinically significant new illness within one month prior to the administration of the drug. A history of severe allergies coronary artery disease, cerebrovascular disease, or a history of syncope; a history of epilepsy or other seizure disorder; Having had an organ transplant; and Having a malignant tumor within 5 years prior to the administration of the above-mentioned drug.

131. The method of any one of claims 105 to 130, wherein the method does not exhibit any property selected from the group consisting of:

132. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, History of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to said administration; 132. The method of any one of claims 105 to 131, wherein the method does not exhibit any property selected from the group consisting of:

133. 133. The method of any one of claims 105-132, wherein the patient's sitting systolic blood pressure is reduced by 0.1 to 15% relative to baseline sitting systolic blood pressure within 12 hours after said administration.

134. 134. The method of any one of claims 105-133, wherein the patient's sitting diastolic blood pressure is reduced by 0.1 to 5% relative to baseline sitting diastolic blood pressure within 6 hours after said administration.

135. 135. The method of 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 improving insulin sensitivity, administered to a patient in need of such treatment, comprising administering to the patient an effective amount of alternatively spliced ​​atrial natriuretic peptide (MANP).

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

138. 137. The method of claim 136, wherein the improvement in insulin sensitivity is a decrease of 0.1 to 0.6 in the patient's updated homeostasis model assessment insulin resistance (HOMA2-IR) value from the baseline HOMA2-IR value within 4 hours after the administration.

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

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

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

142. 140. The method of claim 139, wherein the effective amount is between 6.5 μg and 750 μg.

143. 143. The method of claim 142, wherein the effective amount is between 162.5 μg and 375 μg.

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

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

146. 139. The method of any one of claims 136 to 138, wherein the MANP is administered intravenously followed by subcutaneous administration.

147. 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, followed by subcutaneous administration at a dose of 0.1 μg / kg to 5 μg / kg.

148. 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, followed by subcutaneous administration at a dose of 2.5 μg / kg.

149. 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, followed by subcutaneous administration at a dose of 6.5 μg to 750 μg.

150. 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, followed by subcutaneous administration at a dose of 162.5 μg to 375 μg.

151. The patient is 25 kg / m ２ 151. The method of any one of claims 136 to 150, wherein the patient has a higher body mass index (BMI).

152. The patient is 30 kg / m ２ 152. The method of any one of claims 136 to 151, wherein the patient has a higher BMI.

153. The patient is 35 kg / m ２ 153. The method of any one of claims 136 to 152, wherein the subject has a higher BMI.

154. 154. The method of any one of claims 136-153, wherein the patient exhibits a sitting systolic blood pressure of 140 mmHg to 180 mmHg or a sitting diastolic blood pressure of 90 mmHg to 100 mmHg prior to said administering.

155. 155. The method of any one of claims 136-154, wherein the patient has been taking an antihypertensive drug for 30 days prior to said administering, or has been taking a statin, ezetimibe, or a combination thereof for 60 days prior to said administering.

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

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

158. 158. The method of claim 156 or 157, wherein the elevated serum triglyceride level is identified by a serum triglyceride level of 150 mg / dL or greater in the patient prior to the administration.

159. 159. The method of any one of claims 156 to 158, wherein the low HDL cholesterol level is identified by an HDL cholesterol level of less than 40 mg / dL in a male patient or an HDL cholesterol level of less than 50 mg / dL in a female patient.

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

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

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

163. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to said administration; The patient has received an investigational drug or used an investigational device within 30 days prior to the administration. a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or drug abuse or addiction (within the two years prior to said administration); Having a history of difficulty in donating blood, or having donated blood or blood products within 45 days prior to the administration. The patient has had a clinically significant new illness within one month prior to the administration of the drug. A history of severe allergies coronary artery disease, cerebrovascular disease, or a history of syncope; a history of epilepsy or other seizure disorder; Having had an organ transplant; and Having a malignant tumor within 5 years prior to the administration of the above-mentioned drug.

163. The method of any one of claims 136 to 162, wherein the method does not exhibit any property selected from the group consisting of:

164. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, History of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to said administration; 164. The method of any one of claims 136 to 163, wherein the method does not exhibit any property selected from the group consisting of:

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

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

167. 167. The method of any one of claims 136 to 166, wherein the patient's plasma glucose level is reduced by 1 to 17 mg / mL relative to the baseline plasma glucose level within 4 hours after said administration.

168. The method of 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 acid (NEFA) levels, which is applied to a patient in need of such treatment and comprises administering to the patient an effective amount of alternatively spliced ​​atrial natriuretic peptide (MANP).

170. 170. The method of claim 169, wherein said increasing is an increase of 1 μM in the plasma NEFA level from the baseline NEFA level within 1 hour after said administration.

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

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

173. 173. The method of claim 172, wherein the effective amount is 2.5 μg / kg.

174. 172. The method of claim 171, wherein the effective amount is between 6.5 μg and 750 μg.

175. 175. The method of claim 174, wherein the effective amount is between 162.5 μg and 375 μg.

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

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

178. 171. The method of claim 169 or 170, wherein the MANP is administered intravenously followed by subcutaneous administration.

179. 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, followed by subcutaneous administration at a dose of 0.1 μg / kg to 5 μg / kg.

180. 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, followed by subcutaneous administration at a dose of 2.5 μg / kg.

181. 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, followed by subcutaneous administration at a dose of 6.5 μg to 750 μg.

182. 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, followed by subcutaneous administration at a dose of 162.5 μg to 375 μg.

183. The patient is 25 kg / m ２ 183. The method of any one of claims 169 to 182, wherein the patient has a higher body mass index (BMI).

184. The patient is 30 kg / m ２ 184. The method of any one of claims 169 to 183, wherein the patient has a higher BMI.

185. The patient is 35 kg / m ２ 185. The method of any one of claims 169 to 184, wherein the patient has a higher BMI.

186. 186. The method of any one of claims 169-185, wherein the patient exhibits a sitting systolic blood pressure of 140 mmHg to 180 mmHg or a sitting diastolic blood pressure of 90 mmHg to 100 mmHg prior to said administering.

187. 187. The method of any one of claims 169-186, wherein the patient has been taking an antihypertensive drug for 30 days prior to said administering, or has been taking a statin, ezetimibe, or a combination thereof for 60 days prior to said administering.

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

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

190. 190. The method of claim 188 or 189, wherein the elevated serum triglyceride level is identified by a serum triglyceride level of 150 mg / dL or greater in the patient prior to the administration.

191. 191. The method of any one of claims 188 to 190, wherein the low HDL cholesterol level is identified by an HDL cholesterol level of less than 40 mg / dL in a male patient or an HDL cholesterol level of less than 50 mg / dL in a female patient.

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

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

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

195. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, a history of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to said administration; The patient has received an investigational drug or used an investigational device within 30 days prior to the administration. a history of alcohol abuse, illicit drug use, severe mental illness, physical dependence on any opioid, or drug abuse or addiction (within the two years prior to said administration); Having a history of difficulty in donating blood, or having donated blood or blood products within 45 days prior to the administration. The patient has had a clinically significant new illness within one month prior to the administration of the drug. A history of severe allergies coronary artery disease, cerebrovascular disease, or a history of syncope; a history of epilepsy or other seizure disorder; Having had an organ transplant; and Having a malignant tumor within 5 years prior to the administration of the above-mentioned drug.

195. The method of any one of claims 169 to 194, wherein the method does not exhibit any property selected from the group consisting of:

196. The patient: known hypersensitivity or allergy to MANP or its components, carperitide, other natriuretic peptides, or related compounds; Being pregnant or breastfeeding, History of clinically significant renal parenchymal disease, renal artery stenosis, or fibromuscular dysplasia of the renal artery; and taking a phosphodiesterase-5 inhibitor (sildenafil, vardenafil, or tadalafil) within 72 hours prior to said administration; 196. The method of any one of claims 169 to 195, wherein the method does not exhibit any property selected from the group consisting of:

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

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

199. 200. The method of any one of claims 169 to 198, wherein the patient's plasma glucose level is reduced by 1 to 17 mg / mL relative to the baseline plasma glucose level within 4 hours after said administration.

200. 200. The method of 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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