ACTRII PROTEINS AND USES THEREOF

Abstract

In some aspects, the disclosure relates to compositions and methods comprising ActRII polypeptides for treating, preventing, or reducing the rate of progression and / or severity of pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or combined pulmonary fibrosis (CPFE)), particularly for treating, preventing, or reducing the rate of progression and / or severity of pulmonary hypertension associated with one or more pulmonary diseases associated with co-morbidities (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or combined pulmonary fibrosis (CPFE)).

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of and priority to U.S. Provisional Application No. 63 / 209,871, filed June 11, 2021. The aforementioned application is incorporated herein by reference in its entirety.

[0002] The present application relates to ActRII polypeptides, compositions comprising ActRII polypeptides, and methods for treating, preventing, or reducing the rate of progression and / or severity of pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or combined pulmonary fibrosis (CPFE)), particularly for treating, preventing, or reducing the rate of progression and / or severity of pulmonary hypertension associated with one or more associated pulmonary diseases (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or combined pulmonary fibrosis (CPFE)), which are associated with co-morbid conditions. [Background technology]

[0003] Pulmonary hypertension (PH) is a disease characterized by high blood pressure in the pulmonary vasculature, including the pulmonary arteries, pulmonary veins, and pulmonary capillaries. Generally, PH is defined as a mean pulmonary artery pressure (mPAP) greater than 20 mm Hg at rest or greater than 30 mm Hg during exercise [Hill et al., Respiratory Care 54(7):958-68(2009)]. One of the main symptoms of PH is dyspnea or shortness of breath, while other symptoms include fatigue, dizziness, fainting, peripheral edema (swelling of the feet, legs, or ankles), bluish lips and skin, chest pain, angina, lightheadedness during exercise, nonproductive cough, tachycardia, and palpitations. PH can be a severe disease that leads to heart failure, one of the most common causes of death in people with pulmonary hypertension. Postoperative pulmonary hypertension can complicate many types of surgery or procedures and present challenges associated with a high mortality rate.

[0004] PH can be classified based on the various manifestations of the disease, which share similarities in pathophysiological mechanisms, clinical symptoms, and therapeutic approaches [Simonneau et al., JACC 54(1):S44-54(2009)]. A clinical classification of PH was first proposed in 1973, and a recent updated clinical classification was endorsed by the World Health Organization (WHO) in 2018. According to the updated clinical classification of PH, there are five main groups of PH: pulmonary arterial hypertension (PAH), characterized by a pulmonary artery occlusion pressure (PAWP) <15 mm Hg; PH due to left heart disease (also known as pulmonary venous hypertension or congestive heart failure), characterized by a PAWP >15 mm Hg; PH due to pulmonary disease and / or hypoxia; PH due to pulmonary artery obstruction; and PH of unknown etiology and / or multifactorial etiology [Simonneau et al., JACC 54(1):S44-54(2009); Hill et al., Respiratory Care 54(7):958-68(2009)]. PAH is further classified into idiopathic PAH (IPAH), i.e., a sporadic disease without a family history of PAH or identified risk factors; hereditary PAH; drug- and toxin-induced PAH; PAH associated with connective tissue disease, HIV infection, portal hypertension, congenital heart disease, schistosomiasis, and chronic hemolytic anemia; and persistent PH in newborns [Simonneau et al.,(2019)Eur Respir J:53:1801913]. The diagnosis of the wide variety of PH requires a series of tests.

[0005] In general, PH treatment depends on the cause or classification of PH. When PH is caused by a known medication or medical condition, it is known as secondary PH, and treatment is usually targeted to the underlying disease. Treatment of group 3 pulmonary hypertension has traditionally been to optimize treatment of the underlying lung disease and provide long-term oxygen therapy to those with hypoxia. The effectiveness of pulmonary vasodilators in this patient group is unclear. Furthermore, results from meta-analyses evaluating the effect of vasodilators on exercise tolerance and quality of life are mixed. Although further research is needed to establish which patient groups may benefit most from vasodilator treatment, the current advice is to treat the lungs, not blood pressure. See, for example, McGettrick M.et al.,Glob Cardiol Sci Pract.2020 Apr 30;2020(1).

[0006] There is a high unmet need for effective therapies for treating pulmonary hypertension. Accordingly, it is an object of the present disclosure to provide methods for treating, preventing, or reducing the rate of progression and / or severity of PH, in particular treating, preventing, or reducing the rate of progression and / or severity of one or more PH-related complications. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Hill et al.,Respiratory Care 54(7):958-68(2009) [Non-Patent Document 2] Simonneau et al., JACC 54(1):S44-54(2009) [Non-Patent Document 3] Simonneau et al.,(2019)Eur Respir J:53:1801913 [Non-Patent Document 4] McGettrick M.et al.,Glob Cardiol Sci Pract.2020 Apr 30;2020(1) Summary of the Invention [Means for solving the problem]

[0008] In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a peptide comprising any one of the following amino acids beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191 The present invention provides a method for reducing right ventricular systolic pressure (RVSP) by at least 10%, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 30, 131, 132, 133, 134, or 135.

[0009] In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more pulmonary hypertension-related complications associated with a lung disease, comprising administering to a patient in need thereof a compound or a combination thereof beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 18 , 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135. In some embodiments, the one or more pulmonary hypertension complications associated with the lung disease are selected from the group consisting of persistent cough, productive cough, wheezing, exercise intolerance, respiratory infection, bronchiectasis, chronic infection, nasal polyps, hemoptysis, pneumothorax, respiratory failure, dyspnea, chest pain, hemoptysis, pneumothorax, pulmonary vascular remodeling, pulmonary fibrosis, pulmonary endothelial dysfunction, hypoxia due to chronic lung injury, hypoxic pulmonary vasoconstriction, inflammation, smooth muscle hypertrophy, and right ventricular hypertrophy.

[0010] In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with obstructive pulmonary disease, comprising administering to a patient in need thereof a peptide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362 In one embodiment, the method includes administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 6, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0011] In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with obstructive pulmonary disease, comprising administering to a patient in need thereof a compound or a combination thereof beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 1 In one embodiment, the method includes administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 2, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0012] In some embodiments, the obstructive pulmonary disease is selected from the group consisting of chronic obstructive pulmonary disease (COPD), cystic fibrosis, asthma, emphysema, lymphangioleiomyomatosis, and chronic bronchitis. In some embodiments, the one or more complications of pulmonary hypertension associated with the obstructive pulmonary disease are selected from the group consisting of increased need for supplemental oxygen, decreased mobility, and decreased survival.

[0013] In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with restrictive lung disease, comprising administering to a patient in need thereof a peptide comprising a nucleotide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, In one embodiment, the method includes administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 6, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0014] In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with restrictive lung disease, comprising administering to a patient in need thereof a compound or medicament for use in treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with restrictive lung disease, the compound or medicament for use in treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with restrictive lung disease, the compound or medicament for use in treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with restrictive lung disease, the compound or medicament for use in treating, preventing, or reducing In one embodiment, the method includes administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 2, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0015] In some embodiments, the restrictive lung disease is selected from the group consisting of pulmonary fibrosis, interstitial lung disease, sarcoidosis, idiopathic pulmonary fibrosis, pneumoconiosis, obesity, scoliosis, myasthenia gravis, and pleural effusion. In some embodiments, the one or more complications of pulmonary hypertension associated with restrictive lung disease are selected from the group consisting of shortness of breath on exercise, shortness of breath at rest, shortness of breath with minimal activity, cough, dry cough, wet cough, chronic cough, fatigue, weight loss, anxiety, depression, and fibrosis.

[0016] In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with mixed obstructive and restrictive lung disease, comprising administering to a patient in need thereof a pulmonary hypertension inhibitor (PHI) comprising a pulmonary stimulatory agent (PSA) comprising any one of the following amino acids beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, In one embodiment, the method includes administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 25, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0017] In some embodiments, the mixed obstructive and restrictive lung disease is a pulmonary parenchymal disorder. In some embodiments, the pulmonary parenchymal disorder is selected from the group consisting of sarcoidosis, COPD and ILD, COPD and idiopathic pulmonary fibrosis, pneumoconiosis, ILD, Langerhans cell histiocytosis, IPF, pulmonary alveolar proteinosis, lymphangioleiomyomatosis, and bronchiolitis obliterans syndrome. In some embodiments, the pneumoconiosis is selected from the group consisting of silicosis, coal worker's lung, and beryllium disease. In some embodiments, the ILD is associated with systemic lupus erythematosus, rheumatoid arthritis, connective tissue disease, interstitial pneumonia, stenosing bronchiolitis, or idiopathic organizing pneumonia.

[0018] In some embodiments, the mixed obstructive and restrictive pulmonary disease is a combination of a pulmonary parenchymal disorder and a non-pulmonary disease. In some embodiments, the combination of a pulmonary parenchymal disorder and a non-pulmonary disease is selected from the group consisting of COPD and other non-parenchymal diseases, CHF and other non-pulmonary diseases, asthma and other disorders, ILD and obesity, ILD and CHF, and pulmonary hypoplasia and scoliosis.

[0019] In some embodiments, the COPD and other non-parenchymal diseases are selected from the group consisting of COPD and congestive heart failure (CHF), COPD and obesity, COPD and thoracic surgery, COPD and diaphragmatic paralysis, COPD and scoliosis, and COPD and pleural adhesions. In some embodiments, the CHF and other non-pulmonary diseases are selected from the group consisting of CHF and scoliosis, CHF and lung resection, and CHF and obesity. In some embodiments, the asthma and other disorders are selected from the group consisting of asthma and obesity, asthma and lung resection, asthma and radiation fibrosis, asthma and trapped lung, and asthma and CHF.

[0020] In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with interstitial lung disease (ILD), comprising administering to a patient in need thereof a peptide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, The present invention provides a method for reducing right ventricular systolic pressure (RVSP) by at least 10%, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 29, 130, 131, 132, 133, 134, or 135.

[0021] In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with interstitial lung disease (ILD), comprising administering to a patient in need thereof a compound comprising a nucleotide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 1 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0022] In some embodiments, the ILD is associated with a condition selected from the group consisting of connective tissue disease, sarcoidosis, vascular destruction due to progressive parenchymal fibrosis, vascular inflammation, perivascular fibrosis, thrombotic vascular disease, and endothelial dysfunction. In some embodiments, the connective tissue disease is selected from the group consisting of systemic sclerosis, rheumatoid arthritis, polymositis, dermatomyositis, and Sjogren's syndrome.

[0023] In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), comprising administering to a patient in need thereof a peptide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, In one embodiment, the method includes administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 5, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0024] In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), comprising administering to a patient in need thereof a compound comprising a nucleotide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 1 In one embodiment, the method includes administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 1, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0025] In some embodiments, the one or more complications of pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD) are selected from the group consisting of wheezing, productive cough, frequent cough, chest tightness, shortness of breath without physical activity, shortness of breath with physical activity, respiratory infection, weight loss, leg weakness, leg swelling, and cardiac disease. In some embodiments, the patient has COPD certified by the Global Initiative for Chronic Obstructive Lung Disease as Gold Grade 1, Gold Grade 2, Gold Grade 3, or Gold Grade 4. In some embodiments, the patient has Group A COPD, Group B COPD, Group C COPD, or Group D COPD. In some embodiments, the patient has COPD selected from the group consisting of Stage 1, Stage 2, Stage 3, and Stage 4. In some embodiments, the patient has alpha-1-antitrypsin deficiency.

[0026] In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with pulmonary fibrosis combined with emphysema (CPFE), comprising administering to a patient in need thereof a peptide comprising a nucleotide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 290, 291, 292, 293, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 500, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 600, 601, 602, 603, 604, In one embodiment, the method includes administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 5, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0027] In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with pulmonary fibrosis combined with emphysema (CPFE), comprising administering to a patient in need thereof a compound selected from the group consisting of a nucleotide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 1 In one embodiment, the method includes administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 1, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0028] In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with fibrotic idiopathic interstitial pneumonia (IIP), comprising administering to a patient in need thereof a pulmonary hypertension inhibitor (PHI) or a pulmonary fibrosis inhibitor (PFI ... In one embodiment, the method includes administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 25, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0029] In certain aspects, the disclosure relates to a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with fibrotic idiopathic interstitial pneumonia (IIP), comprising administering to a patient in need thereof a compound or medicament for treating or preventing pulmonary hypertension associated with fibrotic idiopathic interstitial pneumonia (IIP), the ... The methods include administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 21, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135. In some embodiments, the patient has a high fibrosis score and a low carbon monoxide diffusing capacity (DL CO ) one or more diagnostic parameters selected from the group consisting of:

[0030] In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with idiopathic pulmonary fibrosis (IPF), comprising administering to a patient in need thereof a peptide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, , 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0031] In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with idiopathic pulmonary fibrosis (IPF), comprising administering to a patient in need thereof a compound comprising a nucleotide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 18 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135. In some embodiments, the one or more complications of pulmonary hypertension associated with idiopathic pulmonary fibrosis (IPF) are selected from the group consisting of increased need for supplemental oxygen, decreased mobility, and decreased survival.

[0032] In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with non-idiopathic pulmonary fibrosis interstitial lung disease (non-IPF ILD), comprising administering to a patient in need thereof a pulmonary hypertension inhibitor (PHI) or a pulmonary fibrosis ... and administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 8, 129, 130, 131, 132, 133, 134, or 135.

[0033] In certain aspects, the disclosure relates to a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with non-idiopathic pulmonary fibrosis interstitial lung disease (non-IPF ILD), comprising administering to a patient in need thereof a compound comprising an amino acid sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, The present invention provides a method comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 24, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135. In some embodiments, the non-IPF ILD is selected from the group consisting of smoking-related ILD, hypersensitivity pneumonitis-related ILD, connective tissue-associated ILD, occupation-related ILD, and medication-induced ILD. In some embodiments, the one or more complications of pulmonary hypertension associated with non-IPF ILD are selected from the group consisting of increased need for supplemental oxygen, decreased mobility, and decreased survival.

[0034] In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with non-specific interstitial pneumonia (NSIP), comprising administering to a patient in need thereof a peptide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, In one embodiment, the method includes administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 5, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0035] In certain aspects, the disclosure relates to a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with non-specific interstitial pneumonia (NSIP), comprising administering to a patient in need thereof a compound or medicament for treating or preventing pulmonary hypertension associated with non-specific interstitial pneumonia (NSIP), the ... and administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 21, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0036] In some embodiments, the patient has a right ventricular systolic pressure (RVSP) greater than 35 mmHg prior to treatment. In some embodiments, the method reduces the patient's RVSP. In some embodiments, the method reduces the patient's RVSP by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or at least 50%. In some embodiments, the method reduces the patient's RVSP to less than 25 mmHg.

[0037] In some embodiments, the patient has a pulmonary artery systolic pressure (PASP) of greater than 25 mmHg prior to treatment. In some embodiments, the patient has a PASP of at least 35 mmHg, 40 mmHg, 45 mmHg, 50 mmHg, 55 mmHg, or 60 mmHg prior to treatment. In some embodiments, the method reduces the patient's PASP. In some embodiments, the method reduces the patient's PASP by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50%. In some embodiments, the method reduces the patient's PASP by at least 5 mmHg (e.g., at least 5 mmHg, 10 mmHg, 15 mmHg, 20 mmHg, or 25 mmHg). In some embodiments, the method reduces the patient's PASP to less than 25 mmHg. In some embodiments, the method reduces the patient's PASP to less than 20 mmHg.

[0038] In some embodiments, the patient has a pulmonary vascular resistance (PVR) of 3 Wood units or greater prior to treatment. In some embodiments, the method reduces the patient's PVR. In some embodiments, the method reduces the patient's PVR by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%. In some embodiments, the method reduces the PVR to less than 3 Wood units.

[0039] In some embodiments, the patient has a pre-treatment mean pulmonary artery pressure (mPAP) selected from the group consisting of mPAP of at least 17 mmHg, mPAP of at least 20 mmHg, mPAP of at least 25 mmHg, mPAP of at least 30 mmHg, mPAP of at least 35 mmHg, mPAP of at least 40 mmHg, mPAP of at least 45 mmHg, and mPAP of at least 50 mmHg. In some embodiments, the patient has a pre-treatment mPAP of 21-24 mmHg and a pre-treatment PVR of at least 3 Wood units.

[0040] In some embodiments, the patient has a mPAP of greater than 25 mmHg prior to treatment and a cardiac index (CI) of 2.0 L / min / m 2 In some embodiments, the patient has a mPAP of greater than 25 mmHg prior to treatment and a CI of less than 2.5 L / min / m 2 is less than.

[0041] In some embodiments, the method reduces the patient's mPAP by at least 10%, 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50%. In some embodiments, the method reduces the patient's mPAP by at least 3 mmHg, 5, 7, 10, 12, 15, 20, or 25 mmHg. In some embodiments, the method reduces the mPAP to a value selected from the group consisting of less than 17 mmHg, less than 20 mmHg, less than 25 mmHg, and less than 30 mmHg.

[0042] In some embodiments, the patient's pre-treatment mean right atrial pressure (mRAP) is selected from the group consisting of an mRAP of at least 5 mmHg, an mRAP of at least 6 mmHg, an mRAP of at least 8 mmHg, an mRAP of at least 10 mmHg, an mRAP of at least 12 mmHg, an mRAP of at least 14 mmHg, and an mRAP of at least 16 mmHg. In some embodiments, the method improves the patient's mRAP. In some embodiments, the improvement in mRAP is a reduction in mRAP. In some embodiments, the method reduces the patient's mRAP by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%. In some embodiments, the method reduces the patient's mRAP by at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 mmHg.

[0043] In some embodiments, the patient has a cardiac output of less than 4 L / min prior to treatment. In some embodiments, the method increases the patient's cardiac output by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%. In some embodiments, the method increases the patient's cardiac output by at least 0.5 L / min, 1, 1.5, 2, 2.5, 3, 3.5, or 4 L / min in the patient. In some embodiments, the method increases the patient's cardiac output to at least 4 L / min.

[0044] In some embodiments, the patient receives 2.5 L / min / m 2 , 2.0, 1.5, or 1 L / min / m 2 In some embodiments, the method increases the patient's CI by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%. In some embodiments, the method increases the patient's CI by at least 0.2 L / min / m 2 , 0.4, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, or 2 L / min / m 2 In some embodiments, the method increases the patient's CI by at least 2.5 L / min / m 2 Increase to.

[0045] In some embodiments, the method increases the patient's exercise tolerance. In some embodiments, the patient has at least 0.5 index points, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 index points on the Borg Dyspnea Index (BDI) before treatment. In some embodiments, the method reduces the patient's BDI. In some embodiments, the method reduces the patient's BDI by at least 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 index points.

[0046] In some embodiments, the patient has a 6 minute walk distance (6MWD) of less than 550 meters, 500, 450, 440, 400, 380, 350, 300, 250, 200, or 150 meters prior to treatment. In some embodiments, the method increases the patient's 6MWD by at least 10 meters, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, 200, 250, 300, or 400 meters.

[0047] In some embodiments, the method prevents or reduces the progression of a pulmonary hypertension functional classification recognized by the World Health Organization (WHO). In some embodiments, the method prevents or reduces the progression of a pulmonary hypertension functional classification recognized by the WHO from functional class I to class II pulmonary hypertension. In some embodiments, the method prevents or reduces the progression of a pulmonary hypertension functional classification recognized by the WHO from functional class II to class III pulmonary hypertension. In some embodiments, the method prevents or reduces the progression of a pulmonary hypertension functional classification recognized by the WHO from functional class III to class IV pulmonary hypertension. In some embodiments, the method promotes or enhances the regression of a pulmonary hypertension functional classification recognized by the WHO. In some embodiments, the method promotes or enhances the regression of a pulmonary hypertension functional classification recognized by the WHO from class IV to class III pulmonary hypertension. In some embodiments, the method promotes or enhances the regression of a pulmonary hypertension functional classification recognized by the WHO from class III to class II pulmonary hypertension. In some embodiments, the method promotes or enhances the regression of a pulmonary hypertension functional classification recognized by the WHO from class II to class I pulmonary hypertension.

[0048] In some embodiments, the patient has a higher NT-proBNP level than a healthy patient before treatment. In some embodiments, the patient has a normal NT-proBNP level. In some embodiments, the patient has an NT-proBNP level of at least 100 pg / mL, 150, 200, 300, 400, 500, 1000, 3000, 5000, 10,000, 15,000, or 20,000 pg / mL before treatment. In some embodiments, the method reduces the patient's NT-proBNP level. In some embodiments, the method reduces the patient's NT-proBNP level by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or 80%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 30%. In some embodiments, the method reduces NT-proBNP levels to normal levels. In some embodiments, a normal level of NT-proBNP is less than 100 pg / ml.

[0049] In some embodiments, the patient has a higher brain natriuretic peptide (BNP) level than a healthy patient before treatment. In some embodiments, the patient has a normal BNP level before treatment. In some embodiments, the patient has a BNP level of at least 100 pg / mL, 150, 200, 300, 400, 500, 1000, 3000, 5000, 10,000, 15,000, or 20,000 pg / mL before treatment. In some embodiments, the method reduces the patient's BNP level by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or at least 80%. In some embodiments, the method reduces the BNP level to normal levels (i.e., less than 100 pg / ml).

[0050] In some embodiments, the patient has a diastolic pressure gradient (DPG) of greater than 7 mmHg prior to treatment. In some embodiments, the patient has a DPG of at least 7 mmHg (e.g., at least 7, 10, 15, 20, 25, 30, 35, 40, 45, or 50 mmHg) prior to treatment. In some embodiments, the method reduces the patient's DPG. In some embodiments, the method reduces the patient's DPG by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or at least 50%). In some embodiments, the method reduces the patient's DPG to less than 7 mmHg.

[0051] In some embodiments, the method improves the patient's quality of life by at least 1% (e.g., 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%). In some embodiments, the patient's quality of life is measured using the Cambridge Pulmonary Hypertension Outcome Review (CAMPHOR).

[0052] In some embodiments, the patient has pulmonary fibrosis. In some embodiments, the method reduces pulmonary fibrosis in the patient. In some embodiments, the method reduces pulmonary fibrosis in the patient by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%.

[0053] In some embodiments, the patient has a carbon monoxide diffusing capacity (DL) of less than 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, or 20% prior to treatment. CO In some embodiments, the method comprises administering to the patient a DL CO In some embodiments, the method increases the DL COIn some embodiments, the method increases the DL by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%. CO by at least 40%, 45%, 50%, 55%, 60% or 65%.

[0054] In some embodiments, the patient's carbon monoxide transfer coefficient (K CO ) is less than 60% of the predicted value, less than 55% of the predicted value, less than 50% of the predicted value, less than 45% of the predicted value, less than 40% of the predicted value, less than 35% of the predicted value, less than 30% of the predicted value, less than 25% of the predicted value, or less than 20% of the predicted value. In some embodiments, the method comprises: CO In some embodiments, the method increases the patient's K CO In some embodiments, the method increases K by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%. CO by at least 40%, 45%, 50%, 55%, 60% or 65%.

[0055] In some embodiments, the patient's pre-treatment composite physiological index (CPI) is greater than 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, and 80. In some embodiments, the method reduces the patient's CPI. In some embodiments, the method reduces the patient's CPI by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%. In some embodiments, the method reduces the CPI to less than 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, or 5.

[0056] In some embodiments, the patient's pre-treatment arterial oxygen saturation is less than 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, or 30%. In some embodiments, the method increases the patient's arterial oxygen saturation. In some embodiments, the method increases the patient's arterial oxygen saturation by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or at least 50%. In some embodiments, the method increases the arterial oxygen saturation to at least 85%, 90%, or 95%. In some embodiments, the arterial oxygen saturation is measured at rest.

[0057] In some embodiments, the patient's TAPSE is less than 20 mm, 18, 16, 14, or 12 mm. In some embodiments, the method increases TAPSE to at least 20 mm, 22, 24, 26, 28, or 30 mm.

[0058] In some embodiments, the patient's pre-treatment forced vital capacity in one second (FEV1) is selected from the group consisting of greater than 70%, 60%-69%, 50%-59%, 35%-49%, and less than 35%. In some embodiments, the method increases the patient's FEV1. In some embodiments, the method increases the patient's FEV1 by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%. In some embodiments, the method increases the FEV1 by at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%.

[0059] In some embodiments, the patient's pre-treatment forced vital capacity (FVC) is selected from the group consisting of greater than 80%, greater than 70%, 60%-69%, 50%-59%, 35%-49%, and less than 35%. In some embodiments, the method increases the patient's FVC. In some embodiments, the method increases the patient's FVC by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%. In some embodiments, the method increases the FVC by at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%.

[0060] In some embodiments, the method improves right ventricular function in the patient. In some embodiments, the improvement in right ventricular function is due to an increase in right ventricular fractional area change. In some embodiments, the improvement in right ventricular function is due to a decrease in right ventricular hypertrophy. In some embodiments, the improvement in right ventricular function is due to an increase in ejection fraction. In some embodiments, the improvement in right ventricular function is due to an increase in right ventricular fractional area change and ejection fraction. In some embodiments, the method reduces right ventricular hypertrophy in the patient. In some embodiments, the method reduces right ventricular hypertrophy in the patient by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%.

[0061] In some embodiments, the method reduces smooth muscle hypertrophy in a patient. In some embodiments, the method reduces smooth muscle hypertrophy in a patient by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%.

[0062] In some embodiments, the method reduces the risk of death. In some embodiments, the method reduces the risk of death associated with pulmonary arterial hypertension by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%.

[0063] In some embodiments, the method increases transplant-free survival of the patient. In some embodiments, the method increases transplant-free survival of the patient by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%.

[0064] In some embodiments, the method treats one or more comorbidities of pulmonary hypertension associated with a pulmonary disease. In some embodiments, the one or more comorbidities of pulmonary hypertension associated with a pulmonary disease are selected from the group consisting of systemic hypertension, impaired renal function, diabetes mellitus, hyperlipidemia, obesity, coronary artery disease (CAD), obstructive sleep apnea, pulmonary embolism, heart failure, atrial fibrillation, and anemia.

[0065] In some embodiments, the ActRII polypeptide comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence corresponding to residues 30-110 of SEQ ID NO: 1. In some embodiments, the ActRII polypeptide comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:2. In some embodiments, the ActRII polypeptide comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:3.

[0066] In some embodiments, the ActRII polypeptide is a fusion protein further comprising an immunoglobulin Fc domain. In some embodiments, the immunoglobulin Fc domain is an IgG1 immunoglobulin Fc domain. In some embodiments, the Fc fusion protein further comprises a linker domain disposed between the ActRII polypeptide domain and the immunoglobulin Fc domain. In some embodiments, the linker domain is selected from the group consisting of TGGG (SEQ ID NO:20), TGGGG (SEQ ID NO:18), SGGGG (SEQ ID NO:19), GGGGS (SEQ ID NO:22), GGG (SEQ ID NO:16), GGGG (SEQ ID NO:17), and SGGG (SEQ ID NO:21).

[0067] In some embodiments, the ActRII polypeptide comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 23. In some embodiments, the ActRII polypeptide comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 41.

[0068] In some embodiments, the polypeptide comprises an amino acid sequence that is at least 90% identical to an amino acid sequence corresponding to residues 30-110 of SEQ ID NO:1, and the polypeptide binds to activin and / or GDF11. In some embodiments, the polypeptide comprises an amino acid sequence that is at least 90% identical to an amino acid sequence corresponding to residues 21-135 of SEQ ID NO:1, and the polypeptide binds to activin and / or GDF11.

[0069] In some embodiments, the polypeptide is lyophilized. In some embodiments, the polypeptide is soluble. In some embodiments, the polypeptide is administered using subcutaneous injection. In some embodiments, the polypeptide is administered about every 3 weeks. In some embodiments, the polypeptide is administered about every 4 weeks.

[0070] In some embodiments, the polypeptide is part of a homodimeric protein complex. In some embodiments, the polypeptide is glycosylated. In some embodiments, the polypeptide has a glycosylation pattern that can be obtained by expression in Chinese hamster ovary cells.

[0071] In some embodiments, the ActRII polypeptide binds to one or more ligands selected from the group consisting of activin A, activin B, and GDF11. In some embodiments, the ActRII polypeptide further binds to one or more ligands selected from the group consisting of BMP10, GDF8, and BMP6.

[0072] In some embodiments, the ActRII polypeptide is administered at a dose of 0.1 mg / kg to 2.0 mg / kg. In some embodiments, the ActRII polypeptide is administered at a dose of 0.3 mg / kg. In some embodiments, the ActRII polypeptide is administered at a dose of 0.7 mg / kg.

[0073] In some embodiments, the methods disclosed herein further comprise administering to the patient an additional active agent and / or supportive therapy. In some embodiments, the additional active agent and / or supportive therapy is selected from the group consisting of beta-blockers, angiotensin-converting enzyme inhibitors (ACE inhibitors), angiotensin receptor blockers (ARBs), diuretics, lipid-lowering drugs, endothelin blockers, PDE5 inhibitors, and prostacyclin. In some embodiments, the additional active agent and / or supportive therapy is selected from the group consisting of prostacyclin and its derivatives (e.g., epoprostenol, treprostinil, and iloprost); prostacyclin receptor agonists (e.g., selexipag); endothelin receptor antagonists (e.g., thelin, ambrisentan, macitentan, and bosentan); calcium channel blockers (e.g., amlodipine, diltiazem, and nifedipine); anticoagulants (e.g., warfarin); digoxin, diuretics; oxygen therapy; atrial septotomy; pulmonary endarterectomy; phosphodiesterase Type 5 inhibitors (e.g., sildenafil and tadalafil); activators of soluble guanylate cyclase (e.g., cinaciguat and riociguat); ASK-1 inhibitors (e.g., CIIA; SCH79797; GS-4997; MSC2032964A; 3H-naphtho[1,2,3-de]quinyline-2,7-dione, NQDI-1; 2-thioxo-thiazolidine, 5-bromo-3-(4-oxo-2-thioxo-thiazolidine-5-ylidene)-1,3-dihydro-indol-2-one); NF-κB antagonists (e.g., dh404, CDDO-epoxide; 2.2-Difluoropropionamide;C28 imidazole (CDDO-Im);2-Cyano-3,12-dioxoolean-1,9-dien-28-oic acid (CDDO);3-Acetyloleanolic acid;3-Trifluoroacetyloleanolic acid;28-Methyl-3-acetyloleanane;28-Methyl-3-trifluoroacetyloleanane;28-Methyloxyoleanolic acid;SZC014;SCZ015;SZC017;PEGylated derivatives of oleanolic acid;3 -O-(β-D-glucopyranosyl)oleanolic acid;3-O-[β-D-glucopyranosyl-(1-->3)-β-D-glucopyranosyl]oleanolic acid;3-O-[β-D-glucopyranosyl-(1-->2)-β-D-glucopyranosyl]oleanolic acid;3-O-[β-D-glucopyranosyl-(1-->3)-β-D-glucopyranosyl]oleanolic acid 28-O-β-D-glucopyranosyl ester;3-O-[β-D-glucopyranosyl-(1-->2 )-β-D-glucopyranosyl]oleanolic acid 28-O-β-D-glucopyranosyl ester;3-O-[α-L-rhamnopyranosyl-(1-->3)-β-D-glucuronopyranosyl]oleanolic acid;3-O-[α-L-rhamnopyranosyl-(1-->3)-β-D-glucuronopyranosyl]oleanolic acid 28-O-β-D-glucopyranosyl ester;28-O-β-D-glucopyranosyl-oleanolic acid;3-O-β-D-glucopyranosyl(1→3)-β -D-glucopyranosiduronic acid (CS1); oleanolic acid 3-O-β-D-glucopyranosyl(1→3)-β-D-glucopyranosiduronic acid (CS2); methyl 3,11 dioxoolean-12-en-28-oleate (DIOXOL); ZCVI4-2; benzyl 3-dehydro-oxy-1,2,5-oxadiazolo[3',4':2,3]oleanolate); left ventricular assist device (LVAD), oxygen therapy, and lung and / or heart transplantation.

[0074] In some embodiments, the patient is treated with one or more agents selected from the group consisting of phosphodiesterase type 5 inhibitors, soluble guanylate cyclase stimulators, prostacyclin receptor agonists, and endothelin receptor antagonists. In some embodiments, the one or more agents are selected from the group consisting of bosentan, sildenafil, beraprost, macitentan, selexipag, epoprostenol, treprostinil, iloprost, ambrisentan, and tadalafil. In some embodiments, the method further comprises administration of one or more agents selected from the group consisting of phosphodiesterase type 5 inhibitors, soluble guanylate cyclase stimulators, prostacyclin receptor agonists, and endothelin receptor antagonists. In some embodiments, the one or more agents are selected from the group consisting of bosentan, sildenafil, beraprost, macitentan, selexipag, epoprostenol, treprostinil, iloprost, ambrisentan, and tadalafil.

[0075] In some embodiments, the patient is treated with one or more vasodilators prior to administration of the polypeptide. In some embodiments, the method further comprises administering one or more vasodilators. In some embodiments, the one or more vasodilators are selected from the group consisting of prostacyclin, epoprostenol and sildenafil. In some embodiments, the vasodilator is prostacyclin.

[0076] In some embodiments, the patient is receiving one or more therapies for pulmonary hypertension associated with a lung disease. In some embodiments, the one or more therapies for pulmonary hypertension associated with a lung disease are selected from the group consisting of treprostinil, pirfenidone, nintedanib, prostacyclin and its derivatives (e.g., epoprostenol, treprostinil, and iloprost); prostacyclin receptor agonists (e.g., selexipag); endothelin receptor antagonists (e.g., thelin, ambrisentan, macitentan, and bosentan); calcium channel blockers (e.g., amlodipine, diltiazem, and nifedipine); anticoagulants (e.g., warfarin); diuretics; oxygen therapy; atrial septotomy; pulmonary endarterectomy. surgery; phosphodiesterase type 5 inhibitors (e.g., sildenafil and tadalafil); activators of soluble guanylate cyclase (e.g., cinaciguat and riociguat); ASK-1 inhibitors (e.g., CIIA; SCH79797; GS-4997; MSC2032964A; 3H-naphtho[1,2,3-de]quinyline-2,7-dione, NQDI-1; 2-thioxo-thiazolidine, 5-bromo-3-(4-oxo-2-thioxo-thiazolidine-5-ylidene)-1,3-dihydro-indol-2-one); NF-κB antagonists (e.g., dh404, CDDO-epoxide; 2.2-Difluoropropionamide;C28 imidazole (CDDO-Im);2-Cyano-3,12-dioxoolean-1,9-dien-28-oic acid (CDDO);3-Acetyloleanolic acid;3-Trifluoroacetyloleanolic acid;28-Methyl-3-acetyloleanane;28-Methyl-3-trifluoroacetyloleanane;28-Methyloxyoleanolic acid;SZC014;SCZ015;SZC017;PEGylated derivatives of oleanolic acid;3 -O-(β-D-glucopyranosyl)oleanolic acid;3-O-[β-D-glucopyranosyl-(1-->3)-β-D-glucopyranosyl]oleanolic acid;3-O-[β-D-glucopyranosyl-(1-->2)-β-D-glucopyranosyl]oleanolic acid;3-O-[β-D-glucopyranosyl-(1-->3)-β-D-glucopyranosyl]oleanolic acid 28-O-β-D-glucopyranosyl ester;3-O-[β-D-glucopyranosyl-(1-->2 )-β-D-glucopyranosyl]oleanolic acid 28-O-β-D-glucopyranosyl ester;3-O-[α-L-rhamnopyranosyl-(1-->3)-β-D-glucuronopyranosyl]oleanolic acid;3-O-[α-L-rhamnopyranosyl-(1-->3)-β-D-glucuronopyranosyl]oleanolic acid 28-O-β-D-glucopyranosyl ester;28-O-β-D-glucopyranosyl-oleanolic acid;3-O-β-D-glucopyranosyl(1→3)-β -D-glucopyranosiduronic acid (CS1); oleanolic acid 3-O-β-D-glucopyranosyl(1→3)-β-D-glucopyranosiduronic acid (CS2); methyl 3,11 dioxoolean-12-en-28-oleate (DIOXOL); ZCVI4-2; benzyl 3-dehydro-oxy-1,2,5-oxadiazolo[3',4':2,3]oleanolate); left ventricular assist device (LVAD), oxygen therapy, and lung and / or heart transplantation.

[0077] In some embodiments, the ActRII polypeptide is administered to the patient about every week, about every two weeks, about every three weeks, or about every four weeks. In some embodiments, the ActRII polypeptide is administered to the patient every three weeks.

[0078] The file of this patent contains at least one drawing / photograph executed in color. Copies of this patent with color drawing(s) and / or photograph(s) will be provided by the Office upon request and payment of the necessary fee. [Brief description of the drawings]

[0079] [Figure 1] Based on a combined analysis of multiple ActRIIB and ActRIIA crystal structures, an alignment of the extracellular domains of human ActRIIB (sequence number 31) and human ActRIIA (sequence number 2) is shown, including residues predicted herein to directly contact the ligand, as indicated by the boxed outline. [Diagram 2] A multiple sequence alignment of various vertebrate ActRIIA proteins and human ActRIIA (sequence numbers 6-10 and 36-38) is shown. [Diagram 3] Figure 2 shows a multiple sequence alignment of Fc domains from human IgG isotypes using Clustal 2.1. The hinge region is shown with a dotted underline. Double underlines indicate examples of positions engineered in IgG1 Fc (SEQ ID NO: 32) to promote asymmetric chain pairing, as well as corresponding positions for other isotypes IgG2 (SEQ ID NO: 33), IgG3 (SEQ ID NO: 34) and IgG4 (SEQ ID NO: 35). [Figure 4A] 1 shows the purification of ActRIIA-hFc expressed in CHO cells. The protein purifies as a single, well-defined peak, as visualized by a sizing column. [Figure 4B] 1 shows purification of ActRIIA-hFc expressed in CHO cells. The protein is purified as a single, well-defined peak as visualized by Coomassie-stained SDS-PAGE (left lane: molecular weight standards; right lane: ActRIIA-hFc). [Figure 5A] 1 shows binding of ActRIIA-hFc to activin as measured by Biacore™ assay. [Figure 5B] 1 shows binding of ActRIIA-hFc to GDF-11 as measured by Biacore™ assay. [Figure 6A] Shows the effect of ActRIIA-mFc treatment on pulmonary hypertension and RV hypertrophy in the Bleo-MCT PH-ILD rat model. Rx: ActRIIA-mFc sc5mpk, BIW; Bleo: bleomycin; MCT: monocrotaline [Figure 6B] Shows the effect of ActRIIA-mFc treatment on pulmonary hypertension and RV hypertrophy in the Bleo-MCT PH-ILD rat model. Rx: ActRIIA-mFc sc5mpk, BIW; Bleo: bleomycin; MCT: monocrotaline [Figure 6C] Shows the effect of ActRIIA-mFc treatment on pulmonary hypertension and RV hypertrophy in the Bleo-MCT PH-ILD rat model. Rx: ActRIIA-mFc sc5mpk, BIW; Bleo: bleomycin; MCT: monocrotaline [Figure 6D] Shows the effect of ActRIIA-mFc treatment on pulmonary hypertension and RV hypertrophy in the Bleo-MCT PH-ILD rat model. Rx: ActRIIA-mFc sc5mpk, BIW; Bleo: bleomycin; MCT: monocrotaline [Figure 7A] Shows the effect of ActRIIA-mFc treatment on pulmonary hypertension and RV hypertrophy in the Bleo / Su / Hx PH-ILD rat model. Rx: ActRIIA-mFc sc5mpk, BIW; Bleo: bleomycin; MCT: monocrotaline [Figure 7B] Shows the effect of ActRIIA-mFc treatment on pulmonary hypertension and RV hypertrophy in the Bleo / Su / Hx PH-ILD rat model. Rx: ActRIIA-mFc sc5mpk, BIW; Bleo: bleomycin; MCT: monocrotaline [Figure 7C]Shows the effect of ActRIIA-mFc treatment on pulmonary hypertension and RV hypertrophy in the Bleo / Su / Hx PH-ILD rat model. Rx: ActRIIA-mFc sc5mpk, BIW; Bleo: bleomycin; MCT: monocrotaline [Figure 8A] 1 shows the effect of ActRIIA-mFc treatment on Group 3 pulmonary hypertension in an LPS-induced COPD rat model. [Figure 8B] 1 shows the effect of ActRIIA-mFc treatment on Group 3 pulmonary hypertension in an LPS-induced COPD rat model. [Figure 8C] 1 shows the effect of ActRIIA-mFc treatment on Group 3 pulmonary hypertension in an LPS-induced COPD rat model. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0080] 1. Overview The present disclosure relates to compositions and methods for treating pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or combined pulmonary fibrosis and emphysema (CPFE)), comprising administering to a patient in need thereof an effective amount of an ActRII polypeptide described herein. In certain embodiments, the present disclosure provides methods for treating or preventing pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with COPD, ILD, or CPFE) in an individual in need thereof by administering to the individual a therapeutically effective amount of an ActRII polypeptide described herein.

[0081] Most lung diseases can be classified as either obstructive or restrictive. Lung diseases characterized as both obstructive and restrictive occur rarely and are generally caused by a combination of lung parenchymal and nonpulmonary disorders. Obstructive lung diseases (e.g., COPD, chronic bronchitis, asthma, bronchiectasis, bronchiolitis, and cystic fibrosis) are characterized by airway obstruction and are defined by slower, shallower exhalation than in healthy individuals. Restrictive lung diseases (e.g., adult respiratory distress syndrome (ARDS), pneumoconiosis, pneumonia, eosinophilic pneumonia, tuberculosis, sarcoidosis, pulmonary fibrosis and idiopathic pulmonary fibrosis, pleural effusion, and pleurisy) are characterized by a reduction in total lung capacity and are defined by the lungs filling with much less inspiration than would be expected in a healthy individual. One prominent complication of lung disease is pulmonary hypertension. Pulmonary hypertension associated with lung disease (e.g., pulmonary hypertension associated with COPD, ILD, or CPFE) [World Health Organization group 3 PH] is a progressive disease characterized by inflammation and irreversible scarring of lung tissue. Chronic lung disease is the second leading cause of pulmonary hypertension. Patients with pulmonary hypertension associated with lung disease (e.g., pulmonary hypertension associated with COPD, ILD, or CPFE) have been reported to have the highest mortality rate in any of the five diagnostic groups of pulmonary hypertension. Currently, the only treatment approved by the U.S. Food and Drug Administration for pulmonary hypertension associated with lung disease is treprostinil, which is also approved for the treatment of pulmonary arterial hypertension (PAH; WHO group 1 pulmonary hypertension). All other treatments in clinical practice for pulmonary hypertension associated with lung disease are based on management of the underlying lung disease, as well as off-label use of specific treatments approved for pulmonary arterial hypertension (PAH) [World Health Organization (WHO) group 1 PH].

[0082] Pulmonary hypertension associated with lung disease (e.g., pulmonary hypertension associated with COPD, ILD, or CPFE) can be definitively diagnosed using right heart catheterization, but echocardiography remains a good screening and monitoring tool for patients considered at risk. Echocardiography is used to detect elevated pulmonary artery systolic pressure (ePASP) and changes in right ventricular structure or evidence of dysfunction and left heart disease. Other evaluations and / or tools (e.g., 6-minute walk test (6MWT), computed tomography (CT) scan, and pulmonary function tests) may be used. Despite being the only definitive test for pulmonary hypertension associated with lung disease, right heart catheterization is not required for all patients suspected of having the disease. However, right heart catheterization is recommended when moderate or severe pulmonary hypertension is suspected, as well as when another etiology of pulmonary hypertension is suspected.

[0083] In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a pulmonary disease (e.g., obstructive pulmonary disease, restrictive pulmonary disease, or mixed obstructive and restrictive pulmonary disease), comprising administering to a patient in need thereof a pulmonary hypertension inhibitor or pulmonary agonist (e.g., a pulmonary agonist or pulmonary agonist) comprising administering to said patient a pulmonary hypertension inhibitor or pulmonary agonist (e.g., a pulmonary agonist or pulmonary agonist) comprising administering to said patient a pulmonary hypertension inhibitor or pulmonary agonist (e.g., a pulmonary agonist or pulmonary agonist) , 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135. The one or more complications of pulmonary hypertension associated with a lung disease are selected from the group consisting of persistent cough, productive cough, wheezing, exercise intolerance, respiratory infection, bronchiectasis, chronic infection, nasal polyps, hemoptysis, pneumothorax, respiratory failure, dyspnea, chest pain, hemoptysis, pneumothorax, pulmonary vascular remodeling, pulmonary fibrosis, pulmonary endothelial dysfunction, hypoxia due to chronic lung injury, hypoxic pulmonary vasoconstriction, inflammation, smooth muscle hypertrophy, and right ventricular hypertrophy.

[0084] The terms used herein generally have their usual meaning in the art, within the context of this disclosure and within the specific context in which each term is used.Specific terms are discussed below or elsewhere in this specification to provide additional guidance to practitioners in describing the compositions and methods of this disclosure and the methods of making and using them.The scope or meaning of any use of a term will be clear from the specific context in which it is used.

[0085] The term "sequence similarity," in all its grammatical forms, refers to the degree of identity or correspondence between nucleic acid or amino acid sequences that may or may not share a common evolutionary origin.

[0086] "Percent (%) sequence identity" with respect to a reference polypeptide (or nucleotide) sequence is defined as the percentage of amino acid residues (or nucleic acids) in a candidate sequence that are identical to the amino acid residues (or nucleic acids) in the reference polypeptide (nucleotide) sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and does not consider any conservative substitutions as part of the sequence identity. Alignment to determine percent amino acid sequence identity can be accomplished in a variety of ways that are within the skill of the art, for example, using publicly available computer software, such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms necessary to achieve maximum alignment over the entire length of the sequences being compared. However, for purposes herein, % amino acid (nucleic acid) sequence identity values ​​are generated using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program was created by Genentech, Inc., and the source code has been submitted with user documentation to the U.S. Copyright Office, Washington, DC, 20559, where it is registered under U.S. Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, Calif., or may be compiled from the source code. The ALIGN-2 program must be compiled for use on UNIX operating systems, including Digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and do not change.

[0087] "Agonize," in all its grammatical forms, refers to the process of activating a protein and / or gene (e.g., by activating or amplifying gene expression of that protein or by inducing an inactive protein to enter an active state) or increasing the activity of a protein and / or gene.

[0088] "Antagonize," in all its grammatical forms, refers to the process of inhibiting a protein and / or gene (e.g., by inhibiting or reducing gene expression of that protein or by inducing an active protein to enter an inactive state) or decreasing the activity of a protein and / or gene.

[0089] The terms "about" and "approximately", when used in connection with numerical values ​​throughout this specification and claims, indicate an acceptable interval of accuracy familiar to those skilled in the art. Generally, such an interval of accuracy is ±10%. Alternatively, particularly in biological systems, the terms "about" and "approximately" can mean values ​​within an order of magnitude of a given value, preferably within 5-fold, more preferably within 2-fold.

[0090] Numerical ranges disclosed herein are inclusive of the numbers that define the range. The term "between" as used in this application is inclusive of the numbers that define the range. Furthermore, all ranges disclosed herein should be understood to include any and all subranges subsumed therein. For example, a range described as "1 to 10" or "between 1 and 10" should be considered to include any and all subranges that are greater than or equal to 1, such as 1 to 6.1, including a minimum value, and ending at a maximum value of less than or equal to 10, such as 5.5 to 10.

[0091] The terms "a" and "an" include plural referents unless the context in which the term is used clearly dictates otherwise. The terms "a" (or "an"), as well as "one or more" and "at least one" can be used interchangeably herein. Furthermore, as used herein, "and / or" should be interpreted as a specific disclosure of each of two or more specified features or components, regardless of the presence or absence of the other. Thus, the term "and / or" as used herein in phrases such as "A and / or B" is intended to include "A and B," "A or B," "A" (single), and "B" (single). Similarly, the term "and / or" as used in phrases such as "A, B, and / or C" is intended to encompass each of the following aspects: A, B, and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (single); B (single); and C (single).

[0092] Throughout this specification, the word "comprise" or variations such as "comprises" or "comprising" are understood to mean the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

[0093] 2. ActRII Polypeptide In certain aspects, the disclosure relates to ActRII polypeptides and uses thereof for, e.g., treating, preventing, or reducing the rate of progression and / or severity of pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with COPD, ILD, or (CPFE) or one or more complications of pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with COPD, ILD or CPFE). As used herein, the term "ActRII" refers to the family of type II activin receptors. This family includes activin receptor type IIA (ActRIIA) and activin receptor type IIB (ActRIIB).

[0094] In certain embodiments, the present disclosure relates to an ActRII polypeptide having an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in any of SEQ ID NOs: 1, 2, 3, 23, 27, 30, and 41. As used herein, the term "ActRII" refers to the activin receptor type IIA (ActRIIA) protein family, the activin receptor type IIB (ActRIIB) protein family, or combinations and / or variants thereof. ActRII polypeptides can be derived from any species, including variants derived from such ActRII proteins by mutagenesis or other modifications. References to ActRII herein are understood to be references to any one of the currently identified forms. Members of the ActRII family are generally transmembrane proteins composed of a ligand-binding extracellular domain containing a cysteine-rich region, a transmembrane domain, and a cytoplasmic domain with predicted serine / threonine kinase activity.

[0095] The term ActRII polypeptide includes any naturally occurring polypeptide of an ActRII family member, as well as any variant thereof (including mutants, fragments, fusions, and peptidomimetic forms) that retains useful activity. The amino acid numbering of all ActRII-related polypeptides described herein is based on the numbering of the human ActRII precursor protein sequence (SEQ ID NO: 1) provided below, unless otherwise specified.

[0096] The canonical human ActRII precursor protein sequence is as follows:

[0097] TIFF2024521575000002.tif109153TIFF2024521575000003.tif53152The signal peptide is single underlined, the extracellular domain is in bold, and potential endogenous N-linked glycosylation sites are double underlined.

[0098] The processed (mature) extracellular human ActRII polypeptide sequence is as follows:

[0099] TIFF2024521575000004.tif30153The C-terminal "tail" of the extracellular domain is single underlined. The sequence without the "tail" (Δ15 sequence) is as follows:

[0100] The nucleic acid sequence encoding the human ActRII precursor protein is shown below (SEQ ID NO: 4) according to nucleotides 159-1700 of Genbank Reference Sequence NM_001616.4. The signal sequence is underlined.

[0101] TIFF2024521575000006.tif37152TIFF2024521575000007.tif229152TIFF2024521575000008.tif221153The nucleic acid sequence encoding the processed soluble (extracellular) human ActRII polypeptide is as follows:

[0102] TIFF2024521575000009.tif110151 An alignment of the amino acid sequences of the human ActRIIA extracellular domain and the human ActRIIB extracellular domain is shown in Figure 1. This alignment shows the amino acid residues in both receptors that are thought to directly contact the ActRII ligand. For example, the composite ActRII structure showed that the ActRIIA ligand-binding pocket is defined, in part, by residues F31, N33, N35, K38-T41, E47, Y50, K53-K55, R57, H58, F60, T62, K74, W78-N83, Y85, R87, E92, and K94-F101. Conservative mutations are expected to be tolerated at these positions.

[0103] ActRII is well conserved in vertebrates, and large stretches of the extracellular domain are completely conserved. For example, FIG. 2 shows the multiple sequence alignment of the human ActRIIA extracellular domain compared with various ActRIIA orthologues. Many of the ligands that bind to ActRIIA are also highly conserved. Therefore, from these alignments, it is possible to predict the key amino acid positions in the ligand-binding domain that are important for normal ActRII-ligand binding activity, as well as predict the amino acid positions that are likely to tolerate substitution without significantly changing normal ActRII-ligand binding activity. Thus, the active human ActRII variant polypeptides useful according to the method of the present disclosure may contain one or more amino acids from the sequence of another vertebrate ActRII at the corresponding position, or may contain similar residues to those of human or other vertebrate sequences.

[0104] Although not meant to be limiting, the following examples illustrate this approach to defining active ActRII variants. As shown in FIG. 2, F13 of the human extracellular domain is Y in sheep (SEQ ID NO: 7), chicken (SEQ ID NO: 10), cow (SEQ ID NO: 36), barn owl (SEQ ID NO: 37), and bat species (Myotis davidii) (SEQ ID NO: 38) ActRIIA, indicating that aromatic residues including F, W, and Y are tolerated at this position. Q24 of the human extracellular domain is R in cow ActRIIA, indicating that charged residues including D, R, K, H, and E are tolerated at this position. S95 of the human extracellular domain is F in chicken and barn owl ActRIIA, indicating that this site may tolerate a variety of changes, including polar residues such as E, D, K, R, H, S, T, P, G, Y, and possibly hydrophobic residues such as L, I, or F. E52 of the human extracellular domain is D in ovine ActRIIA, indicating that acidic residues including D and E are tolerated at this position. P29 of the human extracellular domain is less conserved and appears as S in ovine ActRIIA and L in bat species (Myotis davidii) ActRIIA, thus essentially any amino acid should be tolerated at this position.

[0105] Moreover, as noted above, ActRII proteins have been characterized in the art in terms of structural / functional properties, particularly with respect to ligand binding [Attisano et al. (1992) Cell 68(1):97-108; Greenwald et al. (1999) Nature Structural Biology 6(1):18-22; Allendorph et al. (2006) PNAS 103(20:7643-7648; Thompson et al. (2003) The EMBO Journal 22(7):1555-1566; and U.S. Patent Nos. 7,709,605, 7,612,041, and 7,842,663]. For example, a defining structural motif is the three-finger toxin fold. The ActRII fold, known as the ActRII fold, is important for ligand binding by type I and type II receptors and is formed by conserved cysteine ​​residues located at various positions within the extracellular domain of each monomeric receptor [Greenwald et al. (1999) Nat Struct Biol 6:18-22; and Hinck (2012) FEBS Lett 586:1860-1870]. In addition to the teachings herein, these references provide ample guidance on how to generate ActRII variants that retain one or more desired activities (e.g., ligand binding activity).

[0106] For example, a defining structural motif, known as the three-finger toxin fold, is important for ligand binding by type I and type II receptors and is formed by conserved cysteine ​​residues located at various positions within the extracellular domain of each monomeric receptor [Greenwald et al. (1999) Nat Struct Biol 6:18-22; and Hinck (2012) FEBS Lett 586:1860-1870]. Thus, the core ligand-binding domain of human ActRII defined by the outermost of these conserved cysteines corresponds to positions 30-110 of SEQ ID NO:1 (ActRII precursor). Thus, the less structurally organized amino acids adjacent to these cysteine-defining core sequences can be truncated by about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 residues at the N-terminus and about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 residues at the C-terminus without necessarily altering ligand binding. Exemplary ActRII extracellular domain truncations include SEQ ID NOs: 2 and 3.

[0107] Thus, the general formula for an active portion (e.g., ligand binding) of ActRII is a polypeptide that comprises, consists essentially of, or consists of amino acids 30-110 of SEQ ID NO: 1. Thus, an ActRII polypeptide can, for example, begin at a residue corresponding to any one of amino acids 21-30 of SEQ ID NO: 1 (e.g., begin at any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) and end at a position corresponding to any one of amino acids 110-135 of SEQ ID NO: 1 (e.g., begin at amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175 22, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134 or 135) or a portion of ActRII that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a portion of ActRII. Other examples include positions selected from 21 to 30 of SEQ ID NO: 1 (e.g., starting with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30), 22 to 30 (e.g., starting with any one of amino acids 22, 23, 24, 25, 26, 27, 28, 29, or 30), 23 to 30 (e.g., starting with any one of amino acids 23, 24, 25, 26, 27, 28, 29, or 30), and 24 to 30 (e.g., starting with any one of amino acids 24, 25, 26, 27, 28, 29, or 30). and ending with any one of amino acids 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135 of SEQ ID NO: 1,134 or 135), 113-135 (e.g., ending with any one of amino acids 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134 or 135), 120-135 (e.g., ending with any one of amino acids 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130 , 131, 132, 133, 134 or 135), 130-135 (e.g., ending with any one of amino acids 130, 131, 132, 133, 134 or 135), 111-134 (e.g., ending with any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133 or 134), 111-133 (e.g., ending with any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, or 133), 111-132 (e.g., ending with any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 132, or 133), 21, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 or 132) or 111-131 (e.g., terminating at any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130 or 131). Variants within these ranges are also specifically contemplated that comprise, consist essentially of, or consist of an amino acid sequence having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the corresponding portion of SEQ ID NO: 1. Thus, in some embodiments,An ActRII polypeptide can comprise, consist essentially of, or consist of a polypeptide that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to amino acids 30-110 of SEQ ID NO: 1. Optionally, an ActRII polypeptide comprises a polypeptide that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to amino acids 30-110 of SEQ ID NO: 1 and includes no more than 1, 2, 5, 10, or 15 conservative amino acid changes in the ligand binding pocket. In some embodiments, the ActRII polypeptide is part of a homodimeric protein complex.

[0108] In certain embodiments, the present disclosure relates to ActRII polypeptides (e.g., ActRIIA polypeptides, ActRIIB polypeptides, or combinations thereof), including fragments, functional variants, and modified forms thereof, and uses thereof (e.g., treating, preventing, or alleviating pulmonary hypertension associated with lung disease, such as pulmonary hypertension associated with COPD, ILD, or CPFE). Preferably, the ActRII polypeptide is soluble (e.g., the extracellular domain of ActRII). In some embodiments, the ActRII polypeptide inhibits (e.g., Smad signaling) one or more GDF / BMP ligands (e.g., GDF11, GDF8, activin A, activin B, GDF3, BMP4, BMP6, BMP10, and / or BMP15). In some embodiments, an ActRII polypeptide binds to one or more GDF / BMP ligands [e.g., GDF11, GDF8, activin A, activin B, GDF3, BMP4, BMP6, BMP10, and / or BMP15]. In some embodiments, an ActRII polypeptide of the disclosure begins at a residue corresponding to amino acids 21-30 of SEQ ID NO:1 (e.g., beginning at any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) and ends at a position corresponding to any one of amino acids 110-135 of SEQ ID NO:1 (e.g., beginning at any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 1, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134 or 135) or a portion of ActRII that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a portion of ActRII.In some embodiments, an ActRII polypeptide comprises, consists of, or consists essentially of an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to amino acids 30-110 of SEQ ID NO: 1. In certain embodiments, an ActRII polypeptide comprises, consists of, or consists essentially of an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to amino acids 21-135 of SEQ ID NO: 1. In some embodiments, the ActRII polypeptide comprises, consists of, or essentially consists of an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOs: 1, 2, 3, 23, 27, 30, and 41.

[0109] In some embodiments, the ActRII polypeptide comprises, consists of, or consists essentially of an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 23. In some alternative embodiments, the ActRII polypeptide (e.g., SEQ ID NO: 23) may lack a C-terminal lysine. In some embodiments, the ActRII polypeptide lacking a C-terminal lysine is SEQ ID NO: 41. In some embodiments, the ActRII polypeptide comprises, consists of, or consists essentially of an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 41. In some embodiments, the patient is administered an ActRII polypeptide that comprises, consists of, or consists essentially of an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:23. In some embodiments, the patient is administered an ActRII polypeptide comprising, consisting of, or consisting essentially of an amino acid sequence at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 41. In some embodiments, the patient is administered a combination of SEQ ID NO: 23 and SEQ ID NO: 41.

[0110] In certain aspects, the present disclosure relates to an ActRII polypeptide (e.g., an ActRIIA polypeptide, an ActRIIB polypeptide, or a combination thereof). In some embodiments, the ActRII trap of the present disclosure is a variant ActRII polypeptide (e.g., an ActRIIA polypeptide, an ActRIIB polypeptide, or a combination thereof), which comprises one or more mutations (e.g., amino acid addition, deletion, substitution, and combinations thereof) in the extracellular domain (also referred to as the ligand-binding domain) of an ActRII polypeptide (e.g., a "wild-type" or unmodified ActRII polypeptide), such that the variant ActRII polypeptide has one or more altered ligand-binding activities compared to the corresponding wild-type ActRII polypeptide. In some embodiments, the variant ActRII polypeptide of the present disclosure retains at least one activity similar to the corresponding wild-type ActRII polypeptide. For example, a preferred ActRII polypeptide binds to and inhibits (e.g., antagonizes) the function of activin, GDF11 and / or GDF8. In some embodiments, the ActRII polypeptides of the present disclosure further bind to and inhibit one or more of the ligands of GDF / BMP (e.g., GDF11, GDF8, activin A, activin B, GDF3, BMP4, BMP6, BMP10, and / or BMP15).Thus, the present disclosure provides ActRII polypeptides with altered binding specificity for one or more ActRII ligands.

[0111] By way of example, one or more mutations can be selected that enhance the selectivity of the altered ligand-binding domain for one or more ActRII binding ligands, e.g., activin (activin A or activin B), particularly GDF11 and / or GDF8 over activin A. Optionally, the K of activin binding at the altered ligand-binding domain can be selected. d K of GDF11 and / or GDF8 binding dThe IC of the altered ligand-binding domain to inhibit activin is at least 2-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, or even 1000-fold greater than the ratio for the wild-type ligand-binding domain. 50 IC for inhibiting GDF11 and / or GDF8 50 The ratio of IC to activin is at least 2-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, or even 1000-fold greater than the wild-type ligand-binding domain. Optionally, the altered ligand-binding domain inhibits GDF11 and / or GDF8 by increasing the IC 50 IC at least 2-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold or even 1000-fold lower than 50 and inhibits.

[0112] In certain embodiments, the present disclosure contemplates specific mutations of an ActRII polypeptide (e.g., an ActRIIA polypeptide, an ActRIIB polypeptide, or a combination thereof) to alter the glycosylation of the polypeptide. Such mutations may be selected to introduce or eliminate one or more glycosylation sites, such as an O-linked or N-linked glycosylation site. An asparagine-linked glycosylation recognition site generally comprises a tripeptide sequence, asparagine-X-threonine or asparagine-X-serine (where "X" is any amino acid), that is specifically recognized by an appropriate cellular glycosylation enzyme. Alterations may also be made by the addition of, or substitution by, one or more serine or threonine residues to the sequence of the polypeptide (for an O-linked glycosylation site). Various amino acid substitutions or deletions at one or both of the first or third amino acid positions (and / or amino acid deletions at the second position) of the glycosylation recognition site result in non-glycosylation in the modified tripeptide sequence. Another means of increasing the number of carbohydrate moieties on a polypeptide is by chemical or enzymatic coupling of glycosides to the polypeptide. Depending on the coupling mode used, the sugar(s) may be attached to (a) arginine and histidine; (b) free carboxyl groups; (c) free sulfhydryl groups, such as those of cysteine; (d) free hydroxyl groups, such as those of serine, threonine or hydroxyproline; (e) aromatic residues, such as those of phenylalanine, tyrosine or tryptophan; or (f) the amide group of glutamine. Removal of one or more carbohydrate moieties present on a polypeptide can be accomplished chemically and / or enzymatically. Chemical deglycosylation can include, for example, exposure of the polypeptide to the compound trifluoromethanesulfonic acid or an equivalent compound. This treatment results in the cleavage of most or all sugars except the linking sugar (N-acetylglucosamine or N-acetylgalactosamine), while leaving the amino acid sequence intact.Enzymatic cleavage of carbohydrate moieties on polypeptides can be achieved by the use of various endo- and exoglycosidases, as described by Thotakura et al. [Meth. Enzymol. (1987) 138:350]. The sequence of the polypeptide may be adjusted accordingly depending on the type of expression system used, since mammalian, yeast, insect, and plant cells can all introduce different glycosylation patterns that can be influenced by the amino acid sequence of the peptide. Generally, the polypeptides of the present disclosure for use in humans can be expressed in mammalian cell lines that provide appropriate glycosylation, such as HEK293 or CHO cell lines, although other mammalian expression cell lines are expected to be useful as well.

[0113] The present disclosure further contemplates a method for generating mutants, particularly a set of combinatorial mutants of ActRII polypeptides (e.g., ActRIIA polypeptides, ActRIIB polypeptides, or combinations thereof), as well as truncation mutants. A pool of combinatorial mutants is particularly useful for identifying functionally active (e.g., GDF / BMP ligand binding) ActRII sequences. The purpose of screening such combinatorial libraries can be to generate polypeptide variants with altered properties, such as, for example, altered pharmacokinetics or altered ligand binding. Various screening assays are provided below, and such assays can be used to evaluate variants. For example, ActRII variants can be screened for their ability to bind to one or more GDF / BMP ligands [e.g., GDF11, GDF8, activin A, activin B, GDF3, BMP4, BMP6, BMP10, and / or BMP15], to prevent binding of GDF / BMP ligands to ActRII polypeptides and their heteromultimers, and / or to disrupt signal transduction triggered by GDF / BMP ligands.

[0114] The activity of ActRII polypeptide (e.g., ActRIIA polypeptide, ActRIIB polypeptide, or combination thereof) or variants thereof can also be tested in cell-based or in vivo assays. For example, the effect of ActRII polypeptide on the expression of genes involved in the pathogenesis of pulmonary hypertension associated with lung disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD) or pulmonary fibrosis with emphysema (CPFE)) can be evaluated. This can be performed in the presence of one or more recombinant ligand proteins [e.g., GDF11, GDF8, activin A, activin B, GDF3, BMP4, BMP6, BMP10, and / or BMP15] as needed, and cells can be transfected to produce ActRII polypeptide and, optionally, GDF / BMP ligand. Similarly, ActRII polypeptide can be administered to mice or other animals, and the effect on the pathogenesis of pulmonary hypertension associated with lung disease (e.g., pulmonary hypertension associated with COPD, ILD or CPFE) can be evaluated using art-recognized methods. Similarly, the activity of ActRII polypeptides or variants thereof can be tested in blood cell progenitor cells for any effect on the proliferation of these cells, for example, by assays described herein and those known in the art. SMAD-responsive reporter genes can be used in such cell lines to monitor effects on downstream signaling.

[0115] Combinatorially inducible variants can be generated that have improved selectivity or generally increased potency compared to a reference ActRII polypeptide (e.g., an ActRIIA polypeptide, an ActRIIB polypeptide, or a combination thereof). Such variants, when expressed from a recombinant DNA construct, can be used in gene therapy protocols. Similarly, mutagenesis can result in variants that have a dramatically different intracellular half-life than the corresponding unmodified ActRII polypeptide. For example, the altered protein can be more stable or less stable to proteolytic degradation or other cellular processes that would otherwise result in the destruction or inactivation of the unmodified polypeptide. Such variants, and the genes that encode them, can be utilized to alter polypeptide complex levels by modulating the polypeptide half-life. For example, a shorter half-life can cause more transient biological effects and, when part of an inducible expression system, can allow for tighter control of recombinant polypeptide complex levels within cells. In Fc fusion proteins, mutations can be made in the linker (if present) and / or the Fc portion to alter the half-life of the ActRII polypeptide.

[0116] Combinatorial libraries can be generated by degenerate libraries of genes that encode a library of polypeptides, each of which contains at least a portion of a potential ActRII polypeptide sequence. For example, a mixture of synthetic oligonucleotides can be enzymatically ligated to a gene sequence such that a degenerate set of potential ActRII-encoding nucleotide sequences can be expressed as individual polypeptides or as a set of larger fusion proteins (e.g., in the case of phage display).

[0117] A library of potential homologues can be generated from degenerate oligonucleotide sequences by many methods. Chemical synthesis of degenerate gene sequences can be performed in an automatic DNA synthesizer, and then the synthetic gene can be ligated into a suitable vector for expression. The synthesis of degenerate oligonucleotides is well known in the art [Narang, SA (1983) Tetrahedron 39:3; Itakura et al. (1981) Recombinant DNA, Proc. 3rd Cleveland Sympos. Macromolecules, ed. AG Walton, Amsterdam: Elsevier pp273-289; Itakura et al. (1984) Annu. Rev. Biochem. 53:323; Itakura et al. (1984) Science 198:1056; and Ike et al. (1983) Nucleic Acid Res. 11:477]. Such techniques have been used in the directed evolution of other proteins [Scott et al., (1990) Science 249:386-390; Roberts et al. (1992) PNAS USA 89:2429-2433; Devlin et al. (1990) Science 249:404-406; Cwirla et al., (1990) PNAS USA 87:6378-6382; and U.S. Patent Nos. 5,223,409, 5,198,346, and 5,096,815].

[0118] Alternatively, other forms of mutagenesis can be used to generate combinatorial libraries. For example, an ActRII polypeptide of the disclosure (e.g., an ActRIIA polypeptide, an ActRIIB polypeptide, or a combination thereof) can be modified by, for example, alanine scanning mutagenesis [Ruf et al. (1994) Biochemistry 33:1565-1572; Wang et al. (1994) J. Biol. Chem. 269:3095-3099; Balint et al. (1993) Gene 137:109-118; Grodberg et al. (1993) Eur. J. Biochem. 218:597-601; Nagashima et al. (1993) J. Biol. Chem. 268:2888-2892; Lowman et al. (1991) Biochemistry 30:10832-10838; and Cunningham et al. (1989) Science 244:1081-1085], by linker scanning mutagenesis [Gustin et al. (1993) Virology 193:653-660; and Brown et al. (1992) Mol. Cell Biol. 12:2644-2652; McKnight et al. (1982) Science 232:316], by saturation mutagenesis [Meyers et al., (1986) Science 232:613], by PCR mutagenesis [Leung et al. (1989) Method Cell Mol Biol 1:11-19], or by random mutagenesis, including chemical mutagenesis [Miller et al. (1992) A Short Course in Bacterial Genetics, CSHL Press, Cold Spring Harbor, NY; and Greener et al. (1994) Strategies in Mol Biol 7:32-34]. Linker scanning mutagenesis is an attractive method for identifying truncated (bioactive) forms of ActRII polypeptides, especially in a combinatorial setting.

[0119] A wide range of techniques are known in the art for screening the gene products of combinatorial libraries made by point mutation and truncation, and even for screening cDNA libraries for gene products with certain properties.Such techniques are generally adaptable to the rapid screening of gene libraries generated by combinatorial mutagenesis of ActRII polypeptides (e.g., ActRIIA polypeptides, ActRIIB polypeptides, or combinations thereof).The most widely used techniques for screening large gene libraries typically include cloning gene libraries into replicable expression vectors, transforming suitable cells with the resulting library of vectors, and expressing combinatorial genes under conditions where detection of desired activity facilitates the relatively easy isolation of the vectors encoding the genes whose products are detected.Exemplary assays include ligand (e.g., GDF11, GDF8, activin A, activin B, GDF3, BMP4, BMP6, BMP10, and / or BMP15) binding assays, and / or ligand-mediated cell signaling assays.

[0120] As will be recognized by those skilled in the art, most of the described mutations, variants or modifications described herein can be made at the nucleic acid level, or in some cases by post-translational modification or chemical synthesis. Such techniques are well known in the art, some of which are described herein. In part, the present disclosure identifies functionally active parts (fragments) and variants of ActRII polypeptides (e.g., ActRIIA polypeptides, ActRIIB polypeptides, or combinations thereof) that can be used as a guide to generate and use other variant ActRII polypeptides within the scope of the disclosure provided herein.

[0121] In certain embodiments, functionally active fragments of the ActRII polypeptide of the present disclosure can be obtained by screening recombinantly produced polypeptides from the corresponding fragments of the nucleic acid encoding the ActRII polypeptide.Furthermore, fragments can be chemically synthesized using techniques known in the art, such as conventional Merrifield solid phase f-Moc or t-Boc chemistry.Fragments can be produced (recombinantly or chemically synthesized) and tested to identify peptidyl fragments that can function as antagonists (inhibitors) of ActRII receptor and / or one or more ligands [e.g., GDF11, GDF8, activin A, activin B, GDF3, BMP4, BMP6, BMP10, and / or BMP15].

[0122] In certain embodiments, the ActRII polypeptides of the present disclosure (e.g., ActRIIA polypeptides, ActRIIB polypeptides, or combinations thereof) may further comprise post-translational modifications in addition to any that naturally occur in the ActRII polypeptide. Such modifications include, but are not limited to, acetylation, carboxylation, glycosylation, phosphorylation, lipidation, and acylation. As a result, the ActRII polypeptides may comprise non-amino acid elements such as polyethylene glycol, lipids, polysaccharides or monosaccharides, and phosphates. The effect of such non-amino acid elements on the functionality of the ligand trap polypeptide can be tested as described herein for other ActRII variants. When the polypeptides of the present disclosure are produced in cells by cleaving the nascent form of the polypeptide, post-translational processing may also be important for the correct folding and / or function of the protein. Different cells (e.g., CHO, HeLa, MDCK, 293, WI38, NIH-3T3 or HEK293) have unique cellular machinery and characteristic mechanisms for such post-translational activities and can be selected to ensure correct modification and processing of ActRII polypeptides.

[0123] In certain aspects, the ActRII polypeptides of the present disclosure (e.g., ActRIIA polypeptides, ActRIIB polypeptides, or combinations thereof) comprise fusion proteins having at least a portion (domain) of an ActRII polypeptide and one or more heterologous portions (domains). Well-known examples of such fusion domains include, but are not limited to, polyhistidine, Glu-Glu, glutathione S-transferase (GST), thioredoxin, protein A, protein G, immunoglobulin heavy chain constant region (Fc), maltose binding protein (MBP) or human serum albumin. Fusion domains can be selected to confer desired properties. For example, some fusion domains are particularly useful for isolating fusion proteins by affinity chromatography. For the purpose of affinity purification, matrices suitable for affinity chromatography are used, such as glutathione, amylase, and nickel or cobalt conjugated resins. Many such matrices are available in "kit" form, for example the Pharmacia GST purification system and the QIAexpress™ system (Qiagen) useful with (HIS6) fusion partners. As another example, the fusion domain may be selected to facilitate detection of the ActRII polypeptide. Examples of such detection domains include various fluorescent proteins (e.g., GFP), as well as "epitope tags," which are usually short peptide sequences for which specific antibodies are available. Well-known epitope tags for which specific monoclonal antibodies are readily available include FLAG, influenza virus hemagglutinin (HA) and c-myc tags. In some cases, the fusion domain has a protease cleavage site, e.g., factor Xa or thrombin, which allows the relevant protease to partially digest the fusion protein, thereby releasing the recombinant protein therefrom. The released protein can then be isolated from the fusion domain by subsequent chromatographic separation.Other types of fusion domains that can be selected include multimerization (e.g., dimerization, tetramerization) domains and functional domains (that confer additional biological function), including, for example, constant domains from immunoglobulins (e.g., Fc domains).

[0124] In certain aspects, the ActRII polypeptides (e.g., ActRIIA polypeptides, ActRIIB polypeptides, or combinations thereof) of the present disclosure include one or more modifications that can "stabilize" the polypeptide. By "stabilize" is meant anything that extends the in vitro half-life, serum half-life, whether this is due to reduced degradation, reduced clearance by the kidney, or other pharmacokinetic effects of the drug. For example, such modifications improve the shelf life of the polypeptide, improve the circulating half-life of the polypeptide, and / or reduce proteolysis of the polypeptide. Such stabilizing modifications include, but are not limited to, fusion proteins (e.g., including fusion proteins that include an ActRII polypeptide domain and a stabilizer domain), modifications of glycosylation sites (e.g., including the addition of glycosylation sites to the polypeptides of the present disclosure), and modifications of carbohydrate moieties (e.g., including the removal of carbohydrate moieties from the polypeptides of the present disclosure). As used herein, the term "stabilizer domain" refers not only to a fusion domain (e.g., an immunoglobulin Fc domain) as in the case of a fusion protein, but also includes non-proteinaceous modifications such as carbohydrate moieties, or non-proteinaceous moieties such as polyethylene glycol. In certain embodiments, an ActRII polypeptide is fused to a heterologous domain (a "stabilizer" domain) that stabilizes the polypeptide, preferably a heterologous domain that increases the stability of the polypeptide in vivo. Fusion with constant domains of immunoglobulins (e.g., Fc domains) is known to confer desirable pharmacokinetic properties to a wide range of proteins. Similarly, fusion to human serum albumin can confer desirable properties.

[0125] An example of a naturally occurring amino acid sequence that may be used for the Fc portion of human IgG1 (G1Fc) is shown below (SEQ ID NO: 11). The dotted underline indicates the hinge region and the solid underline indicates the position that contains a naturally occurring variant. In part, the disclosure provides a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence having 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 11. Naturally occurring variants in G1Fc would include E134D and M136L according to the numbering system used in SEQ ID NO: 11 (see Uniprot P01857).

[0126] TIFF2024521575000010.tif77151Optionally, the IgG1 Fc domain has one or more mutations at residues such as Asp-265, lysine 322 and Asn-434. In certain cases, the mutant IgG1 Fc domain has one or more of these mutations (e.g., Asp-265 mutation) and has a reduced ability to bind to Fcγ receptors compared to wild-type Fc domains. In other cases, the mutant Fc domain has one or more of these mutations (e.g., Asn-434 mutation) and has an increased ability to bind to MHC class I-related Fc receptors (FcRNs) compared to wild-type IgG1 Fc domains.

[0127] An example of a native amino acid sequence that may be used for the Fc portion of human IgG2 (G2Fc) is shown below (SEQ ID NO:12). The dotted underline indicates the hinge region and the double underline indicates a position in the sequence where there is a database discrepancy (from UniProt P01859). In part, the disclosure provides a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence having 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO:12.

[0128] TIFF2024521575000011.tif77151 Two examples of amino acid sequences that can be used for the Fc portion of human IgG3 (G3Fc) are shown below. The hinge region of G3Fc can be up to four times longer than other Fc chains and contains three identical 15-residue segments preceded by a similar 17-residue segment. The first G3Fc sequence shown below (SEQ ID NO: 13) contains a short hinge region consisting of a single 15-residue segment, while the second G3Fc sequence (SEQ ID NO: 14) contains a full-length hinge region. In both cases, the dotted underline indicates the hinge region and the solid underline indicates the position that contains the naturally occurring variant according to UniProt P01859. In part, the disclosure provides polypeptides comprising, consisting essentially of, or consisting of an amino acid sequence having 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NOs:13 and 14.

[0129] TIFF2024521575000012.tif37151TIFF2024521575000013.tif133151Naturally occurring variants of G3Fc (see, for example, Uniprot P01860) include E68Q, P76L, E79Q, Y81F, D97N, N100D, T124A, S169N, S169del, F221Y when converted to the numbering system used in SEQ ID NO: 13, and the present disclosure provides fusion proteins that include a G3Fc domain that contains one or more of these variants. Also, the human immunoglobulin IgG3 gene (IGHG3) shows structural polymorphisms characterized by different hinge lengths (see Uniprot P01860). Specifically, variant WIS lacks most of the V region and all of the CH1 region. It has an extra interchain disulfide bond at position 7 in addition to the 11 normally present in the hinge region. Variant ZUC lacks most of the V region, all of the CH1 region, and part of the hinge. Variant OMM may represent an allelic form or another gamma chain subclass. The present disclosure provides additional fusion proteins that include a G3Fc domain containing one or more of these variants.

[0130] An example of a naturally occurring amino acid sequence that may be used for the Fc portion of human IgG4 (G4Fc) is shown below (SEQ ID NO:15). The dotted underline indicates the hinge region. In part, the disclosure provides a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence having 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO:15.

[0131] Various engineered mutations in the TIFF2024521575000014.tif77151 Fc domain are presented herein with respect to the G1Fc sequence (SEQ ID NO: 11), and similar mutations in G2Fc, G3Fc, and G4Fc can be derived from their alignment with G1Fc in Figure 3. Because of the different hinge lengths, similar Fc positions based on isotype alignment (Figure 3) have different amino acid numbers in SEQ ID NOs: 11, 12, 13, 14, and 15. The hinge region, C H 2 Area and C H A given amino acid position in a three-region immunoglobulin sequence (e.g., SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, and SEQ ID NO:15) is numbered as in the Uniprot database (C H 1, hinge, C H 2 and C H It will be understood that when including the entire IgG1 heavy chain constant domain (consisting of three regions), the same positions are identified by different numbers. For example, the human G1Fc sequence (SEQ ID NO: 11), the human IgG1 heavy chain constant domain (Uniprot P01857) and selected Cs in the human IgG1 heavy chain H The correspondence between the three positions is as follows: [Table 1]

[0132] Various methods are known in the art to increase the desired pairing of Fc-containing fusion polypeptide chains in a single cell line to produce asymmetric fusion proteins in acceptable yields [Klein et al (2012) mAbs 4:653-663; and Spiess et al (2015) Molecular Immunology 67(2A):95-106]. Methods for obtaining the desired pairing of Fc-containing chains include, but are not limited to, charge-based pairing (electrostatic steering), "knobs-into-holes" steric pairing, SEEDbody pairing, and leucine zipper-based pairing [Ridgway et al (1996) Protein Eng 9:617-621; Merchant et al (1998) Nat Biotech 16:677-681; Davis et al (2010) Protein Eng Des Sel 23:195-202; Gunasekaran et al (2010); 285:19637-19646; Wranik et al (2012) J Biol Chem 285:19637-19646]. 287:43331-43339; U.S. Pat. No. 5,932,448; WO 1993 / 011162; WO 2009 / 089004, and WO 2011 / 034605].

[0133] It is understood that the various elements of a fusion protein (e.g., an immunoglobulin Fc fusion protein) can be arranged in any manner consistent with desired functionality. For example, an ActRII polypeptide domain can be arranged C-terminally toward a heterologous domain, or a heterologous domain can be arranged C-terminally toward an ActRII polypeptide domain. The ActRII polypeptide domain and the heterologous domain do not need to be adjacent in the fusion protein, and additional domains or amino acid sequences can be included C-terminally or N-terminally toward either domain, or between domains.

[0134] For example, an ActRII receptor fusion protein can include an amino acid sequence shown in formula ABC. The B portion corresponds to an ActRII polypeptide domain (e.g., an ActRIIA polypeptide, an ActRIIB polypeptide, or a combination thereof). The A portion and the C portion can be independently zero, one, or more amino acids, and both the A portion and the C portion, if present, are heterologous to the B portion. The A portion and / or the C portion can be linked to the B portion via a linker sequence. The linker can be glycine-rich (e.g., 2-10, 2-5, 2-4, 2-3 glycine residues) or rich in glycine and proline residues, and can include, for example, a single sequence of threonine / serine and glycine or a repeat sequence of threonine / serine and / or glycine, such as GGG (SEQ ID NO: 16), GGGG (SEQ ID NO: 17), TGGGG (SEQ ID NO: 18), SGGGG (SEQ ID NO: 19), TGGG (SEQ ID NO: 20), SGGG (SEQ ID NO: 21), or GGGGS (SEQ ID NO: 22) singlet, or repeat. In certain embodiments, an ActRII fusion protein comprises an amino acid sequence set forth in the formula ABC, where A is a leader (signal) sequence, B consists of an ActRII polypeptide domain, and C is a polypeptide moiety that improves one or more of in vivo stability, in vivo half-life, uptake / administration, tissue localization or distribution, protein complex formation, and / or clearance. In certain embodiments, an ActRII fusion protein comprises an amino acid sequence set forth in the formula ABC, where A is a TPA leader sequence, B consists of an ActRII receptor polypeptide domain, and C is an immunoglobulin Fc domain. Exemplary fusion proteins comprise the amino acid sequence set forth in any one of SEQ ID NOs: 23, 27, 30, and 41.

[0135] In some embodiments, the ActRII polypeptide used according to the methods described herein is an isolated polypeptide. As used herein, an isolated protein or polypeptide is one that is separated from the components of its natural environment. In some embodiments, the polypeptide of the present disclosure is purified to more than 95%, 96%, 97%, 98% or 99% purity, for example, as measured by electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reverse phase HPLC). Methods for assessing purity are well known in the art [see, for example, Flatman et al., (2007) J. Chromatogr. B 848:79-87]. In some embodiments, the ActRII polypeptide used according to the methods described herein is a recombinant polypeptide.

[0136] The ActRII polypeptides of the present disclosure can be produced by various techniques known in the art. For example, the polypeptides of the present disclosure can be synthesized using standard protein chemistry techniques, such as those described in Bodansky, M. Principles of Peptide Synthesis, Springer Verlag, Berlin (1993), and Grant GA (ed.), Synthetic Peptides: A User's Guide, WH Freeman and Company, New York (1992). In addition, automated peptide synthesizers are commercially available (e.g., Advanced ChemTech Model 396; Milligen / Biosearch 9600). Alternatively, the polypeptides of the present disclosure, including their fragments or variants, can be produced recombinantly using various expression systems well known in the art (e.g., E. coli, Chinese hamster ovary (CHO) cells, COS cells, baculovirus). In further embodiments, modified or unmodified polypeptides of the present disclosure can be produced by digestion of recombinantly produced full-length ActRII polypeptides, for example, by using proteases such as trypsin, thermolysin, chymotrypsin, pepsin, or paired basic amino acid converting enzymes (PACE). Computer analysis (using commercially available software, for example, MacVector, Omega, PCGene, Molecular Simulation, Inc.) can be used to identify proteolytic cleavage sites. Alternatively, such polypeptides can be produced from recombinantly produced full-length ActRII polypeptides using chemical cleavage (e.g., cyanogen bromide, hydroxylamine, etc.).

[0137] 3. Nucleic acids encoding ActRII polypeptides In certain embodiments, the disclosure provides isolated and / or recombinant nucleic acids encoding ActRII polypeptides (e.g., ActRIIA polypeptides, ActRIIB polypeptides, or combinations thereof), including fragments, functional variants, and fusion proteins thereof.

[0138] As used herein, isolated nucleic acid(s) refers to a nucleic acid molecule that has been separated from a component of its natural environment. Isolated nucleic acid includes a nucleic acid molecule contained in a cell that ordinarily contains the nucleic acid molecule, but where the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0139] In certain embodiments, nucleic acids encoding the ActRII polypeptides of the disclosure are understood to include nucleic acids that are variants of any one of SEQ ID NOs: 4, 5, or 28. Variant nucleotide sequences include sequences that differ by one or more nucleotide substitutions, additions, or deletions, including allelic variants, and thus include a coding sequence that differs from the nucleotide sequence set forth in any one of SEQ ID NOs: 4, 5, or 28.

[0140] In certain embodiments, the ActRII polypeptides of the present disclosure are encoded by isolated and / or recombinant nucleic acid sequences that are at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of SEQ ID NOs: 4, 5, or 28. One of skill in the art will appreciate that nucleic acid sequences that are at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to a sequence complementary to SEQ ID NO: 4, 5, or 28, and variants thereof, are also within the scope of the present disclosure. In further embodiments, the nucleic acid sequences of the present disclosure may be isolated, recombinant, and / or fused to a heterologous nucleotide sequence, or in a DNA library.

[0141] In other embodiments, the nucleic acid of the present disclosure also includes nucleotide sequences that hybridize under high stringency conditions to the nucleotide sequences set forth in SEQ ID NO: 4, 5, or 28, the complementary sequences of SEQ ID NO: 4, 5, or 28, or fragments thereof. As mentioned above, one skilled in the art will readily understand that suitable stringency conditions that promote DNA hybridization can be varied. One skilled in the art will readily understand that suitable stringency conditions that promote DNA hybridization can be varied. For example, hybridization can be performed at about 45°C in 6.0× sodium chloride / sodium citrate (SSC), followed by washing at 50°C in 2.0× SSC. For example, the salt concentration in the washing step can be selected from low stringency of about 2.0× SSC at 50°C to high stringency of about 0.2× SSC at 50°C. In addition, the temperature of the washing step can be increased from low stringency conditions of room temperature and about 22°C to high stringency conditions of about 65°C. Both temperature and salt may be varied, or temperature or salt concentration may be held constant while the other variable is varied. In one embodiment, the nucleic acids provided in this disclosure are hybridized under low stringency conditions of 6×SSC at room temperature, followed by washing with 2×SSC at room temperature.

[0142] Also within the scope of this disclosure are isolated nucleic acids that differ from the nucleic acids set forth in SEQ ID NO: 4, 5 or 28 depending on the degeneracy of the genetic code. For example, some amino acids are specified by more than one triplet. Codons that specify the same amino acid or synonyms (e.g., CAU and CAC are synonyms for histidine) may result in "silent" mutations that do not affect the amino acid sequence of the protein. However, DNA sequence polymorphisms that result in changes in the amino acid sequence of the subject proteins are expected to exist among mammalian cells. One of skill in the art will understand that these mutations in one or more nucleotides (up to about 3-5% of the nucleotides) of a nucleic acid encoding a particular protein may exist in individuals of a given species due to natural allelic variation. All such nucleotide variations and resulting amino acid polymorphisms are within the scope of this disclosure.

[0143] In certain embodiments, the recombinant nucleic acid of the present disclosure may be operably linked to one or more regulatory nucleotide sequences in an expression construct. The regulatory nucleotide sequence is generally appropriate for the host cell used for expression. Numerous types of suitable expression vectors and suitable regulatory sequences are known in the art and can be used in a variety of host cells. Typically, the one or more regulatory nucleotide sequences may include, but are not limited to, a promoter sequence, a leader or signal sequence, a ribosome binding site, transcriptional start and stop sequences, translational start and stop sequences, and an enhancer or activator sequence. Constitutive or inducible promoters known in the art are contemplated by the present disclosure. The promoter may be either a naturally occurring promoter or a hybrid promoter that combines elements of two or more promoters. The expression construct may be present in the cell on an episome, such as a plasmid, or the expression construct may be inserted into a chromosome. In some embodiments, the expression vector contains a selectable marker gene to allow for the selection of transformed host cells. Selectable marker genes are well known in the art and may vary with the host cell used.

[0144] In certain embodiments, the nucleic acid of the subject matter disclosed herein is provided in an expression vector comprising a nucleotide sequence encoding an ActRII polypeptide (e.g., an ActRIIA polypeptide, an ActRIIB polypeptide, or a combination thereof) operably linked to at least one regulatory sequence. Regulatory sequences are art-recognized and are selected to direct the expression of an ActRII polypeptide. Thus, the term regulatory sequence includes promoters, enhancers, and other expression control elements. Exemplary regulatory sequences are described in Goeddel; Gene Expression Technology: Methods in Enzymology, Academic Press, San Diego, CA (1990). For example, any of a wide variety of expression control sequences that control the expression of a DNA sequence can be used in these vectors to express a DNA sequence encoding an ActRII polypeptide when operably linked. Such useful expression control sequences include, for example, the SV40 early and late promoters, the tet promoter, the adenovirus or cytomegalovirus immediate early promoters, the RSV promoter, the lac system, the trp system, the TAC or TRC system, the T7 promoter whose expression is directed by the T7 RNA polymerase, the major operator and promoter regions of phage lambda, the control region of the fd coat protein, the promoter of 3-phosphoglycerate kinase or other glycolytic enzymes, the promoter of acid phosphatase, e.g., Pho5, the promoter of yeast alpha mating factor, the polyhedron promoter of the baculovirus system, and other sequences known to control the expression of genes in prokaryotic or eukaryotic cells or their viruses, and various combinations thereof. It will be understood that the design of the expression vector may depend on such factors as the choice of the host cell to be transformed and / or the type of protein desired to be expressed. Additionally, the copy number of the vector, the ability to control that copy number, and the expression of any other proteins encoded by the vector, such as antibiotic markers, should also be considered.

[0145] The recombinant nucleic acid of the present disclosure can be produced by ligating cloned gene or a part thereof into a vector suitable for expression in prokaryotic cells, eukaryotic cells (yeast, avian, insect or mammalian cells) or both.The expression vehicle for producing recombinant ActRII polypeptide includes plasmids and other vectors.For example, suitable vectors include the following types of plasmids: pBR322-derived plasmids, pEMBL-derived plasmids, pEX-derived plasmids, pBTac-derived plasmids, and pUC-derived plasmids for expression in prokaryotic cells, such as E. coli.

[0146] Some mammalian expression vectors contain both prokaryotic sequences to facilitate propagation of the vector in bacteria and one or more eukaryotic transcription units that are expressed in eukaryotic cells. pcDNAI / amp, pcDNAI / neo, pRc / CMV, pSV2gpt, pSV2neo, pSV2-dhfr, pTk2, pRSVneo, pMSG, pSVT7, pko-neo and pHyg-derived vectors are examples of mammalian expression vectors suitable for transfection of eukaryotic cells. Some of these vectors are modified with sequences from bacterial plasmids such as pBR322 to facilitate replication and drug resistance selection in both prokaryotic and eukaryotic cells. Alternatively, derivatives of viruses such as bovine papillomavirus (BPV-1) or Epstein-Barr virus (pHEBo, pREP-derived and p205) can be used for transient expression of proteins in eukaryotic cells. Examples of other viral (including retroviral) expression systems can be found below in the description of gene therapy delivery systems. The various methods used for preparing plasmids and transforming host organisms are well known in the art. Other suitable expression systems for both prokaryotic and eukaryotic cells, as well as general recombinant procedures, can be found, for example, in Molecular Cloning A Laboratory Manual, 3rd Ed., ed. by Sambrook, Fritsch and Maniatis (Cold Spring Harbor Laboratory Press, 2001). In some instances, it may be desirable to express recombinant polypeptides by using baculovirus expression systems. Examples of such baculovirus expression systems include pVL-derived vectors (e.g., pVL1392, pVL1393 and pVL941), pAcUW-derived vectors (e.g., pAcUW1), and pBlueBac-derived vectors (e.g., β-gal containing pBlueBac III).

[0147] In one embodiment, the vector will be, for example, the Pcmv-Script vector (Stratagene, La Jolla, Calif.), the pcDNA4 vector (Invitrogen, Carlsbad, Calif.), and the pCI-neo vector (Promega, Madison, Wisc.) designed for production of a subject ActRII polypeptide in CHO cells. As will be apparent, the subject genetic constructs can be used to express a subject ActRII polypeptide in cells grown in culture, e.g., to produce proteins, including fusion proteins or variant proteins, for purification.

[0148] The present disclosure also relates to a host cell transfected with a recombinant gene comprising one or more coding sequences of the subject ActRII polypeptide.The host cell can be any prokaryotic or eukaryotic cell.For example, the ActRII polypeptide of the present disclosure can be expressed in bacterial cells such as E. coli, insect cells (e.g., using baculovirus expression system), yeast, or mammalian cells (e.g., Chinese hamster ovary (CHO) cell line).Other suitable host cells are known to those skilled in the art.

[0149] Thus, the present disclosure further relates to a method for producing the subject ActRII polypeptide. For example, a host cell transfected with an expression vector encoding an ActRII polypeptide can be cultured under appropriate conditions to cause expression of the ActRII polypeptide. The polypeptide can be secreted and isolated from a mixture of cells and medium containing the polypeptide. Alternatively, the ActRII polypeptide can be retained by the cytoplasm or in a membrane fraction, the cells can be harvested, lysed, and the protein isolated. The cell culture comprises host cells, medium, and other by-products. Suitable media for cell culture are well known in the art. The subject polypeptide can be isolated from cell culture medium, host cells, or both using techniques known in the art for purifying proteins, including ion exchange chromatography, gel filtration chromatography, ultrafiltration, electrophoresis, immunoaffinity purification with an antibody specific for a particular epitope of the ActRII polypeptide, and affinity purification with an agent that binds to a domain fused to the ActRII polypeptide (e.g., a protein A column can be used to purify an ActRII-Fc fusion protein). In some embodiments, the ActRII polypeptide is a fusion protein that contains a domain that facilitates its purification.

[0150] In some embodiments, purification is achieved by a series of column chromatography steps, including, for example, three or more of the following in any order: Protein A chromatography, Q Sepharose chromatography, phenyl sepharose chromatography, size exclusion chromatography, and cation exchange chromatography. Purification can be completed by virus filtration and buffer exchange. ActRII protein can be purified to a purity of more than 90%, more than 95%, more than 96%, more than 98% or more than 99% as determined by size exclusion chromatography, and more than 90%, more than 95%, more than 96%, more than 98% or more than 99% as determined by SDS PAGE. The target level of purity must be sufficient to achieve the desired results in mammalian systems, particularly non-human primates, rodents (mouse) and humans.

[0151] In another embodiment, a fusion gene encoding a purification leader sequence, e.g., a poly-(His) / enterokinase cleavage site sequence at the N-terminus of the desired portion of the recombinant ActRII polypeptide, is 2+ The expressed fusion protein can be purified by affinity chromatography using metal resin.The purification leader sequence can then be subsequently removed by treatment with enterokinase to obtain purified ActRII polypeptide.See, for example, Hochuli et al.(1987) J. Chromatography 411:177; and Janknecht et al.(1991) PNAS USA 88:8972.

[0152] The technique for making fusion genes is well known. Essentially, the linking of various DNA fragments encoding different polypeptide sequences is carried out according to conventional techniques using blunt or sticky ends for ligation, restriction enzyme digestion to provide suitable termini, filling of cohesive ends as necessary, alkaline phosphatase treatment to avoid undesired ligation, and enzymatic ligation. In another embodiment, fusion genes can be synthesized by conventional techniques, including automatic DNA synthesizers. Alternatively, PCR amplification of gene fragments can be carried out using anchor primers that generate complementary overhangs between two consecutive gene fragments and can then be annealed to generate chimeric gene sequences. For example, see Current Protocols in Molecular Biology, eds.Ausubel et al., John Wiley&Sons:1992.

[0153] 4.How to use In part, the disclosure relates to a method of treating pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)), the method comprising administering to a patient in need thereof an effective amount of an ActRII polypeptide as described herein. In some embodiments, the disclosure relates to a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)), the method comprising administering to a patient in need thereof an effective amount of an ActRII polypeptide as described herein. In some embodiments, the ActRII polypeptide is administered at a dose range of 0.1 mg / kg to 2.0 mg / kg (e.g., 0.3 mg / kg or 0.7 mg / kg). In some embodiments, administration of an ActRII polypeptide results in a change in one or more hemodynamic or functional parameters (e.g., a decrease in pulmonary vascular resistance (PVR); an increase in 6-minute walk distance (6MWD); a decrease in N-terminal pro-B-type natriuretic peptide (NT-proBNP) levels; prevention or delay of progression of a World Health Organization (WHO)-recognized pulmonary hypertension functional class; promotion or enhancement of regression of a WHO-recognized pulmonary hypertension functional class; improvement in right ventricular function; and improvement in pulmonary artery pressure).

[0154] In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a peptide comprising any one of the following amino acids beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191 The present invention relates to a method for reducing right ventricular systolic pressure (RVSP) by at least 10%, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 30, 131, 132, 133, 134, or 135.

[0155] In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a peptide or peptides that begin with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 290, 291, 292, 293, 294, 295, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352 In one embodiment, the method comprises administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135. In some embodiments, the one or more pulmonary hypertension complications associated with the lung disease are selected from the group consisting of persistent cough, productive cough, wheezing, exercise intolerance, respiratory infection, bronchiectasis, chronic infection, nasal polyps, hemoptysis, pneumothorax, respiratory failure, dyspnea, chest pain, hemoptysis, pneumothorax, pulmonary vascular remodeling, pulmonary fibrosis, pulmonary endothelial dysfunction, hypoxia due to chronic lung injury, hypoxic pulmonary vasoconstriction, inflammation, smooth muscle hypertrophy, and right ventricular hypertrophy.

[0156] These methods are particularly aimed at therapeutic and prophylactic treatment of animals, more specifically humans. The terms "subject", "individual" or "patient" are used interchangeably throughout this specification and refer to either human or non-human animals. These terms include mammals, such as humans, non-human primates, laboratory animals, livestock animals (including cows, pigs, camels, etc.), companion animals (e.g., dogs, cats, other domestic animals, etc.) and rodents (e.g., mice and rats). In certain embodiments, the patient, subject or individual is a human.

[0157] The terms "treatment," "treating," "alleviating," "reducing the rate of progression," "reducing the severity," and the like are used herein to generally mean obtaining a desired pharmacological and / or physiological effect, and may also be used to refer to improving, alleviating, and / or reducing the severity of one or more clinical complications of the condition being treated (e.g., pulmonary hypertension associated with lung disease). The effect may be preventative in that it completely or partially delays the onset or recurrence of the disease, condition, or its complications, and / or may be therapeutic in that it partially or completely cures the disease or condition and / or adverse effects caused by the disease or condition. As used herein, "treatment" encompasses any treatment of a disease or condition in a mammal, particularly a human. As used herein, a therapeutic agent that "prevents" a disorder or condition refers to a compound that, in a statistical sample, reduces the occurrence of the disorder or condition in a treated sample compared to an untreated control sample, or delays the onset of the disease or condition compared to an untreated control sample.

[0158] In general, the treatment or prevention of a disease or condition described in the present disclosure (e.g., pulmonary hypertension associated with lung disease) is achieved by administering one or more ActRII polypeptides of the present disclosure in an "effective amount". An effective amount of an agent refers to an amount effective at the dosage and for the period of time necessary to achieve a desired therapeutic or preventive result. The "therapeutically effective amount" of an agent of the present disclosure may vary according to factors such as the disease state, age, sex, and weight of an individual, as well as the ability of the agent to induce a desired response in an individual. A "prophylactically effective amount" refers to an amount effective at the dosage and for the period of time necessary to achieve a desired preventive result.

[0159] In certain aspects, the present disclosure contemplates the use of an ActRII polypeptide in combination with one or more additional active agents or other supportive therapies to treat or prevent a disease or condition (e.g., pulmonary hypertension associated with lung disease). As used herein, "combination with administration," "combination of administration," "combined with administration," or "conjoint administration" refers to any form of administration in which the additional active agent or supportive therapy (e.g., second, third, fourth, etc.) is still effective in the body (e.g., multiple compounds are effective in a patient simultaneously for a period of time, and may include synergistic effects of the compounds). Efficacy may not correlate with measurable concentrations of the agent in blood, serum, or plasma. For example, different therapeutic compounds can be administered either in the same formulation or in separate formulations, simultaneously or sequentially, and on separate schedules. Thus, subjects undergoing such treatment can benefit from the combined effects of the different active agents or therapies. One or more ActRII polypeptides of the present disclosure can be administered simultaneously with, before, or after one or more other additional agents or supportive therapies, such as those disclosed herein. Generally, each active agent or therapy will be administered at a dose and / or time schedule determined for that particular agent. The particular combination to use in the regimen will take into account compatibility of the ActRII polypeptide of the present disclosure with the additional active agent or therapy, and / or the desired effect.

[0160] Overview of WHO classification Pulmonary hypertension conditions treated by the methods described herein can include any one or more of the conditions recognized according to the World Health Organization (WHO). See, e.g., Simonneau (2019) Eur Respir J:53:1801913. [Table 2] TIFF2024521575000017.tif170150

[0161] As used herein, the term "pulmonary hemodynamic parameters" refers to any parameter used to describe or evaluate blood flow through the heart and pulmonary vasculature. Examples of pulmonary hemodynamic parameters include, but are not limited to, mean pulmonary artery pressure (mPAP), diastolic pulmonary artery pressure (dPAP) [also known as pulmonary artery diastolic pressure (PADP)], systolic pulmonary artery pressure (sPAP) [also known as pulmonary artery systolic pressure (PASP)], mean right atrial pressure (mRAP), pulmonary capillary wedge pressure (PCWP) [also known as pulmonary artery wedge pressure (PAWP)], pulmonary vascular resistance (PVR) and cardiac output (CO).

[0162] Many of the above pulmonary hemodynamic parameters are interrelated. For example, PVR is related to mPAP, PCWP and CO according to the following formula: PVR=(mPAP-PCWP) / CO [Wood units].

[0163] PVR measures the resistance to flow imposed by the pulmonary vasculature without the influence of left-sided filling pressure. PVR may also be measured according to the following formula:

[0164] PVR=TPG×80 / CO [unit: dyne-second-cm -5 ] or PVR = (mPAP-PCWP) x 80 / CO [unit: dyne-seconds-cm -5 ] In some embodiments, the total PVR can be measured using the following formula:

[0165] TPR=mPAP / CO In some embodiments, a normal PVR is between 20 and 180 dyne sec-cm -5 , or typically less than 0.5 to 2 Wood units. According to some embodiments, elevated PVR can refer to a PVR of more than 2 Wood units, more than 2.5 Wood units, more than 3 Wood units, or more than 3.5 Wood units.

[0166] As yet another example, mPAP is related to dPAP and sPAP according to the following formula: mPAP=(2 / 3)dPAP+(1 / 3)sPAP.

[0167] In some embodiments, the pulmonary hemodynamic parameters are measured directly, for example during right heart catheterization, while in other embodiments, the pulmonary hemodynamic parameters are estimated and / or assessed by other techniques, such as magnetic resonance imaging (MRI) or echocardiography.

[0168] Exemplary pulmonary hemodynamic parameters include mPAP, PAWP and PVR. One or more pulmonary hemodynamic parameters can be measured by any suitable procedure, for example, by utilizing right heart catheterization or echocardiography. Various hemodynamic characteristics of PH and pulmonary hypertension associated with pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD) or pulmonary fibrosis with emphysema (CPFE)) are shown in Table 2. [Table 3]

[0169] The clinical classification or hemodynamic characteristics and associated diagnostic parameters of PAH described herein may be updated or modified based on the availability of new or existing data sources or when additional clinical entities are considered.

[0170] Characteristics of Pulmonary Hypertension Associated with Lung Disease Pulmonary hypertension associated with lung disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD) or combined pulmonary fibrosis (CPFE)) (WHO group 3 PH) is the second most common form of pulmonary hypertension and is associated with increased morbidity and mortality. Patients with group 3 pulmonary hypertension have a poorer outcome than patients with group 1 pulmonary hypertension. Similarly, patients with group 1 pulmonary arterial hypertension and associated lung disease suffer even worse outcomes compared to patients with group 1 pulmonary arterial hypertension alone.

[0171] Various factors contribute to the pathogenesis of pulmonary hypertension associated with lung disease. These factors vary based on the underlying lung disease. For example, in pulmonary hypertension caused by COPD, the most prominent pathogenesis of pulmonary hypertension is hypoxic pulmonary vasoconstriction (HPVC) with remodeling of the pulmonary vascular bed. Early changes during vascular remodeling include distal neomuscularization of arterioles, intimal thickening, and medial hypertrophy. This remodeling ultimately leads to reduced vascularity and, as a result, increased peripheral vascular resistance seen in pulmonary hypertension. Additional mechanisms underlying ILD-associated pulmonary hypertension include vascular destruction due to progressive parenchymal fibrosis, vascular inflammation, perivascular fibrosis, thrombotic vasculopathy, and endothelial dysfunction. More specifically, patients with pulmonary hypertension associated with idiopathic pulmonary fibrosis (IPF) may have an abnormal vascular phenotype characterized by abnormal gene expression profiles that promote vascular remodeling.

[0172] Pulmonary hypertension associated with lung disease can be diagnosed with a mean pulmonary artery pressure (mPAP) greater than 25 mmHg. Pulmonary hypertension associated with lung disease can result in persistent cough, productive cough, wheezing, exercise intolerance, respiratory infection, bronchiectasis, chronic infection, nasal polyps, hemoptysis, pneumothorax, respiratory failure, dyspnea, chest pain, hemoptysis, pneumothorax, pulmonary vascular remodeling, pulmonary fibrosis, pulmonary endothelial dysfunction, hypoxia due to chronic lung injury, hypoxic pulmonary vasoconstriction, inflammation, smooth muscle hypertrophy, and right ventricular hypertrophy. Pulmonary hypertension associated with lung disease can be classified as either obstructive or restrictive lung disease. Obstructive lung disease (e.g., COPD, cystic fibrosis, asthma, emphysema, and chronic bronchitis) is characterized by difficulty in breathing. Alternatively, restrictive lung diseases can be further divided into intrinsic (e.g., pulmonary fibrosis, interstitial lung disease, sarcoidosis, idiopathic pulmonary fibrosis) and extrinsic (obesity, scoliosis, myasthenia gravis, and pleural effusion) disorders, and are characterized by limitation of complete lung expansion.

[0173] Pulmonary hypertension associated with lung disease (e.g., pulmonary hypertension associated with lung disease) can be difficult to diagnose due to the heterogeneity of the underlying lung conditions. Many symptoms of lung disease are similar to those of pulmonary hypertension. However, there are some clinical features that prompt the diagnosis of pulmonary hypertension associated with lung disease (e.g., exertional dyspnea or hypoxemia not adequately explained by parenchymal lung disease or sleep disorders, rapid decline in arterial oxygenation during exercise, any clinical features suggestive of right heart failure, pulmonary artery enlargement, attenuation of the peripheral pulmonary vasculature, or right ventricular enlargement as shown by high-resolution computed tomography (HRCT), severely reduced diffusing capacity as shown by pulmonary function tests, and lung biopsy). Klings, ES (2021). Pulmonary hypertension due to lung disease and / or hypoxemia (group 3 pulmonary hypertension): Epidemiology, pathogenesis, and diagnostic evaluation in adults. UpToDate. Retrieved April 6, 2021, from https: / / www.uptodate.com / contents / pulmonary-hypertension-due-to-lung-disease-and-or-hypoxemia-group-3-pulmonary-hypertension-epidemiology-pathogenesis-and-diagnostic-evaluation-in-adults.

[0174] Although echocardiography is the standard test when investigating patients with suspected pulmonary hypertension with unknown underlying pulmonary disease and / or sleep-disordered breathing etiology, echocardiography may be unreliable to accurately diagnose pulmonary hypertension in patients with severe pulmonary disease. In such cases, right heart catheterization (RHC) can provide a more accurate assessment.

[0175] Chronic obstructive pulmonary disease In some embodiments, the disclosure provides a method of treating pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), comprising administering to a patient in need thereof an effective amount of a pulmonary hypertension inhibitor (PHI) or ... The present invention relates to a method comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135. Chronic obstructive pulmonary disease (also known as chronic obstructive pulmonary disease (COPD)) is an inflammatory lung disease that causes obstruction of airflow from the lungs. Among this group of diseases are emphysema and chronic bronchitis. According to the Centers for Disease Control and Prevention, millions of people suffer from COPD, 16 million of whom are in the United States.

[0176] The severity of COPD is determined using the Global Initiative for Chronic Obstructive Lung Disease (GOLD) staging or grading system, which is determined by spirometry results (GOLD 1: mild, GOLD 2: moderate, GOLD 3: severe, GOLD 4: very severe). This system determines the stage of COPD based on several factors (e.g., overall symptoms, number of COPD exacerbations, hospitalizations due to COPD exacerbations, and spirometry results). The majority of patients with pulmonary hypertension caused by COPD present with severe or very severe airflow obstruction (GOLD spirometry stage 3 or 4, FEV-1 <50% of predicted) or severe emphysema, and mild to moderate precapillary pulmonary hypertension. Current treatments for COPD include short-acting bronchodilators, long-acting bronchodilators, inhaled steroids, both bronchodilators and inhaled steroids or combination inhalers containing multiple bronchodilators, oral steroids, phosphodiesterase-4 inhibitors, theophylline, antibiotics, various types of pulmonary therapies, and home noninvasive ventilation.

[0177] Interstitial lung disease In some embodiments, the disclosure provides a method of treating pulmonary hypertension associated with interstitial lung disease (ILD), comprising administering to a patient in need thereof a compound comprising any one of the following amino acids beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, 10, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, The present invention relates to a method for treating a lung disease comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 25, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135. Interstitial lung disease or ILD is a chronic lung disease that occurs as a result of damage between the air sacs of the lungs, causing lung scarring, inflammation, and breathing problems. ILD can be caused by infection, medicines, and inhalation of harmful particles in the air. The underlying cause of ILD determines the course of treatment. ILD, overall, can reduce the quality of life of a person living with the disease and shorten the person's lifespan overall.

[0178] There are approximately five categories of ILD based on their underlying cause: exposure- or occupation-related ILD (e.g., asbestosis, silicosis, hypersensitivity pneumonitis), ILD associated with drug and / or medical treatment (e.g., chemotherapy, radiation therapy), ILD associated with autoimmune disorders and / or connective tissue diseases (e.g., lupus, scleroderma, polymyositis or dermatomyositis, rheumatoid arthritis), sarcoidosis, and idiopathic ILD. Outside the five general categories, there are ILDs such as idiopathic pulmonary fibrosis (IPF), bronchiolitis obliterans, histiocytosis X, chronic eosinophilic pneumonia, collagen vascular disease, granulomatous vasculitis, Goodpasture's syndrome, and pulmonary alveolar proteinosis.

[0179] Although the symptoms of ILD can vary from person to person, as well as based on the particular ILD, a common thread between the various forms of ILD is that all ILDs begin with inflammation of the bronchioles (e.g., bronchiolitis), alveoli (e.g., alveolitis), or capillaries (vasculitis). The most common symptoms of ILD, such as shortness of breath (especially with exercise), fatigue and weakness, loss of appetite, weight loss, dry cough that does not produce phlegm, chest discomfort, labored breathing, and bleeding in the lungs, may mimic other lung conditions or medical problems.

[0180] Fibrosis leads to the continued destruction of the air sacs, inter- and peri-air sac lung tissue, and pulmonary capillaries. Disease progression can be gradual or rapid, and symptoms can be very mild, moderate, or very severe. The course of ILD is unpredictable, but can be improved by medical intervention.

[0181] Interstitial lung disease is diagnosed using pulmonary function tests (PFTs), chest x-rays, blood tests (e.g., analysis of arterial blood gases to determine the amount of carbon dioxide and oxygen in the blood), high-resolution CT (HRCT, CT, or CAT scan), bronchoscopy, bronchoalveolar lavage, and lung biopsy.

[0182] Treatment plans for ILD are typically determined based on an individual's age, overall health, and medical history, the extent of the disease, the individual's tolerance to certain medications, procedures, and / or therapies, expectations for the course of the disease, and the individual's opinions or preferences. These treatment plans may include oral medications (e.g., corticosteroids), supplemental oxygen, and lung transplantation.

[0183] Idiopathic pulmonary fibrosis and other idiopathic interstitial pneumonias In some embodiments, the disclosure provides a method of treating pulmonary hypertension associated with idiopathic pulmonary fibrosis (IPF), comprising administering to a patient in need thereof a peptide comprising any one of the peptides beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, 10, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 3 25, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135. Idiopathic pulmonary fibrosis (IPF) (also idiopathic fibrosing alveolitis, chronic idiopathic fibrosing alveolitis, interstitial pneumonia) is one of the most frequently diagnosed interstitial lung diseases (ILDs), affecting approximately 13-20 per 100,000 people worldwide, with 30,000-40,000 new cases diagnosed each year. Although medical treatments for IPF are available, the disease remains serious and clinical deterioration is expected.

[0184] There are several underlying factors that influence the progression of IPF, one of which is thought to be chronic and / or recurrent microinjury of the alveolar epithelium (e.g., exposure to environmental pollutants, acid aspiration from gastroesophageal reflux, and viral infections). Damage to the epithelium is followed by injury and / or activation of cells lining the vascular and interstitial compartments of the lung, the epithelium of the distal airways, and resident macrophages. Genetic factors may contribute to IPF, as suggested by the occurrence of IPF-like disease in patients with rare genetic disorders, and cases of familial idiopathic interstitial pneumonia.

[0185] Currently, there are no treatments that have proven effective in halting the progression of the disease, although newer medications (e.g., pirfenidone and nintedanib) have been approved by the Food and Drug Administration to help slow the progression of the disease.

[0186] Nonidiopathic pulmonary fibrosis interstitial lung disease In some embodiments, the disclosure provides a method of treating pulmonary hypertension associated with non-idiopathic pulmonary fibrosis interstitial lung disease (non-IPF ILD), comprising administering to a patient in need thereof a compound beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, The present invention relates to a method for treating a lung disease comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 28, 129, 130, 131, 132, 133, 134, or 135. Non-idiopathic pulmonary fibrosis interstitial lung disease (non-IPF) causes inflammation and fibrosis of the lung interstitium, resulting in impaired gas exchange. The estimated prevalence of non-IPF ranges from 25 to 74 per 100,000 people. There are more than 200 known causes of non-IPF, which can typically be classified as occupational and environmental exposures, organic substances causing hypersensitivity pneumonitis, drug-induced pulmonary toxicity, connective tissue disorders, and systemic diseases.

[0187] The pathogenesis of non-IPF is similar among non-IPFs that arise from any one of more than 200 known causes, including a stage of injury (e.g., recurrent and direct epithelial / endothelial injury to distal airspaces and disruption of the alveolar-capillary basement membrane), inflammation caused by release of proinflammatory cytokines and chemokines (e.g., transforming growth factor-β) by macrophages, and repair (e.g., formation of myofibroblasts and secretion of fibrous proteins and substrates that form the extracellular matrix). However, repetition of this process over time leads to thickening of the lung parenchyma and continued irreversible fibrosis.

[0188] Treatment and management of the disease includes supportive care, supplemental oxygen, and, in certain conditions, corticosteroids. Lung transplantation may be considered as an option in severe or progressive cases. Mortality can be as high as 100% during acute exacerbations of non-IPF.

[0189] Emphysema with pulmonary fibrosis In some embodiments, the disclosure provides a method of treating pulmonary hypertension associated with pulmonary fibrosis combined with emphysema (CPFE), comprising administering to a patient in need thereof a peptide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, The present invention relates to a method comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135. Pulmonary fibrosis with emphysema (CPFE) is characterized by dyspnea, upper lobe emphysema, lower lobe pulmonary fibrosis, and abnormalities in gas exchange. CPFE can be further complicated by pulmonary hypertension, acute lung injury, and lung cancer. CPFE is diagnosed using pulmonary function tests (PFTs) that are different from those used to diagnose fibrosis or emphysema alone. Additionally, HRCT scans can be used to detect the concomitant occurrence of emphysema and pulmonary fibrosis.

[0190] CPFE is associated with smoking, exposure to asbestos and mineral dust, hypersensitivity pneumonitis (or farmer's lung), and being male, and has a significant mortality rate. Median survival times range from 2.1 to 8.5 years, and if pulmonary hypertension is present, the 1-year survival rate is only 60%. Despite this disparity, there is no specific treatment for CPFE other than supportive care (e.g., smoking cessation).

[0191] Diagnosis of Pulmonary Hypertension Associated with Lung Disease The diagnosis of pulmonary hypertension associated with lung disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD) or pulmonary hypertension associated with emphysema complicated by pulmonary fibrosis (CPFE)) can be determined based on symptoms and physical examination, using a review of a comprehensive set of parameters to determine whether functional groups, hemodynamic and other criteria are met. Some of the criteria that may be considered include the patient's clinical symptoms (e.g., shortness of breath, fatigue, weakness, angina, syncope, dry cough, exercise-induced nausea and vomiting), electrocardiogram (ECG) results, chest radiograph results, pulmonary function tests, arterial blood gases, echocardiography results, ventilation / perfusion lung scan results, high-resolution CT results, contrast-enhanced CT results, pulmonary angiography results, cardiac magnetic resonance imaging, blood tests (e.g., biomarkers such as BNP or NT-proBNP), immunology, abdominal ultrasound scan, right heart catheterization (RHC), vascular reactivity, and genetic testing. Galie N.,et al Euro Heart J. (2016) 37, 67-119. The diagnosis of pulmonary hypertension associated with lung disease (Group 3 pulmonary hypertension) is determined when a person with pulmonary hypertension has chronic lung disease and / or hypoxemia and no alternative causes of pulmonary hypertension can be identified. Group 3 pulmonary hypertension can be based on clinical evaluation and echocardiographic findings and can be definitively confirmed by right heart catheterization. Pulmonary hypertension associated with lung disease overlaps symptomatically and etiologically with other types of pulmonary hypertension, but several features distinguish this group from others (e.g., moderate to severe impairment (FEV1<60% in COPD patients and FVC<70% in pulmonary fibrosis patients), characteristic imaging or polysomnographic findings of pulmonary impairment, reduced expiratory reserve volume, normal oxygen pulses, mixed venous oxygen saturation above the lower limit of normal, and elevated arterial partial pressure of carbon dioxide during exercise (especially in COPD), as well as the presence of mild to moderate pulmonary hypertension on echocardiography or right heart catheterization).

[0192] Group 3 pH readings Various pulmonary hemodynamic parameters are useful in assessing disease progression and patient responsiveness to treatment protocols. Typically, these parameters describe or evaluate blood flow through the heart and pulmonary vasculature. Examples of pulmonary hemodynamic parameters include, but are not limited to, mean pulmonary artery pressure (mPAP), diastolic pulmonary artery pressure (dPAP) [also known as pulmonary artery diastolic pressure (PADP)], systolic pulmonary artery pressure (sPAP) [also known as pulmonary artery systolic pressure (PASP)], mean right atrial pressure (mRAP), pulmonary capillary wedge pressure (PCWP) [also known as pulmonary artery wedge pressure (PAWP)], pulmonary vascular resistance (PVR) and cardiac output (CO). In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a peptide comprising a nucleotide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 1 , 130, 131, 132, 133, 134, or 135, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of the following: a) It reduces right ventricular systolic pressure (RVSP); b) Decrease mPAP c) reduce mRAP; d) Decreasing PVR; e) Decrease the diastolic pressure gradient (DPG); f) decreasing BNP levels; g) reducing NT-proBNP levels; h) reduces smooth muscle hypertrophy; i) reduce the patient's CAMPHOR score; j) improve ventricular function; k) reduce right ventricular hypertrophy; l) Increases cardiac index; m) Increase cardiac output; n) reducing complex physiological indicators; o) Increases arterial oxygen saturation; p) Increase exercise tolerance; q) Increase forced expiratory volume; r) Increase forced vital capacity (FVC); s)DL CO Increase; t) reducing pulmonary fibrosis; and / or u) Increase transplant-free survival of patients.

[0193] mPAP Pulmonary blood pressure is usually much lower than systemic blood pressure. Normal pulmonary artery pressure is typically between 8 and 20 mmHg at rest. When pressure in the pulmonary arteries is greater than 25 mmHg at rest or greater than 30 mmHg during physical activity, it is abnormally high and is characterized as pulmonary hypertension.

[0194] In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a compound or medicament for use in treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, the ... The present invention relates to a method of lowering mPAP in a patient by at least 10%, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 25, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135. In some embodiments, the method relates to a patient having an mPAP of at least 17 mmHg. In some embodiments, the method relates to a patient having an mPAP of at least 20 mmHg. In some embodiments, the method relates to a patient having an mPAP of at least 25 mmHg. In some embodiments, the method relates to a patient having an mPAP of 25-34 mmHg. In some embodiments, the method relates to patients with an mPAP of at least 30 mmHg. In some embodiments, the method relates to patients with an mPAP of at least 35 mmHg. In some embodiments, the method relates to patients with an mPAP of at least 40 mmHg. In some embodiments, the method relates to patients with an mPAP of at least 45 mmHg. In some embodiments, the method relates to patients with an mPAP of at least 50 mmHg.

[0195] In some embodiments, the method relates to patients with an mPAP of 21-24 mmHg and a PVR of at least 3 Wood units. In some embodiments, the method relates to patients with an mPAP of greater than 25 mmHg and a cardiac index (CI) of 2.0 L / min / m 2In some embodiments, the method relates to patients having an mPAP of greater than 25 mmHg and a CI of less than 2.5 L / min / m 2 For patients who are less than

[0196] In some embodiments, the methods relate to reducing the patient's mPAP by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or at least 50%). In some embodiments, the methods relate to reducing the patient's mPAP by at least 15%. In some embodiments, the methods relate to reducing the patient's mPAP by at least 20%. In some embodiments, the methods relate to reducing the patient's mPAP by at least 25%. In some embodiments, the methods relate to reducing the patient's mPAP by at least 30%. In some embodiments, the methods relate to reducing the patient's mPAP by at least 35%. In some embodiments, the methods relate to reducing the patient's mPAP by at least 40%. In some embodiments, the methods relate to reducing the patient's mPAP by at least 45%. In some embodiments, the methods relate to reducing the patient's mPAP by at least 50%.

[0197] In some embodiments, the method relates to lowering mPAP in the patient by at least 3 mmHg. In some embodiments, the method relates to lowering mPAP by at least 5 mmHg. In some embodiments, the method relates to lowering mPAP by at least 7 mmHg. In some embodiments, the method relates to lowering mPAP by at least 10 mmHg. In some embodiments, the method relates to lowering mPAP by at least 12 mmHg. In some embodiments, the method relates to lowering mPAP by at least 15 mmHg. In some embodiments, the method relates to lowering mPAP by at least 20 mmHg. In some embodiments, the method relates to lowering mPAP by at least 25 mmHg. In some embodiments, the method relates to lowering mPAP to less than 17 mmHg. In some embodiments, the method relates to lowering mPAP to less than 20 mmHg. In some embodiments, the method relates to lowering mPAP to less than 25 mmHg. In some embodiments, the method relates to lowering mPAP to less than 30 mmHg.

[0198] mRAP Right atrial pressure (RAP) is the blood pressure in the right atrium of the heart. RAP reflects the amount of blood returning to the heart and the heart's ability to pump blood into the arterial system. Normal right atrial pressure is typically between 2 mmHg and 6 mmHg. Elevated right atrial pressure reflects right ventricular (RV) loading and is an established risk factor.

[0199] In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a compound or medicament for use in treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, the ... The present invention relates to a method for reducing mRAP in a patient by at least 10%, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 25, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0200] In some embodiments, the patient has a mean right atrial pressure (mRAP) of at least 5 mmHg. In some embodiments, the patient has a mean right atrial pressure (mRAP) of at least 6 mmHg. In some embodiments, the patient has a mean right atrial pressure (mRAP) of at least 8 mmHg. In some embodiments, the patient has a mean right atrial pressure (mRAP) of at least 10 mmHg. In some embodiments, the patient has a mean right atrial pressure (mRAP) of at least 12 mmHg. In some embodiments, the patient has a mean right atrial pressure (mRAP) of at least 14 mmHg. In some embodiments, the patient has a mean right atrial pressure (mRAP) of at least 16 mmHg. In some embodiments, the method improves the patient's mean right atrial pressure (mRAP). In some embodiments, the improvement in mRAP is a reduction in mRAP.

[0201] In some embodiments, the method reduces the patient's mRAP by at least 10%. In some embodiments, the method reduces the patient's mRAP by at least 15%. In some embodiments, the method reduces the patient's mRAP by at least 20%. In some embodiments, the method reduces the patient's mRAP by at least 25%. In some embodiments, the method relates to reducing the patient's mRAP by at least 30%. In some embodiments, the method relates to reducing the patient's mRAP by at least 35%. In some embodiments, the method relates to reducing the patient's mRAP by at least 40%. In some embodiments, the method relates to reducing the patient's mRAP by at least 45%. In some embodiments, the method relates to reducing the patient's mRAP by at least 50%.

[0202] In some embodiments, the method reduces mRAP by at least 1 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 1 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 2 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 3 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 4 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 5 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 6 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 7 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 8 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 9 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 10 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 11 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 12 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 13 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 14 mmHg. In some embodiments, the method reduces mRAP in a patient by at least 15 mmHg.

[0203] PVR Vascular resistance is the resistance that must be overcome to push blood through the circulatory system and create flow. Pulmonary vascular resistance is the resistance to blood flow from the pulmonary artery to the left atrium. Total blood flow refers to cardiac output (5-6 L / min). Normal values ​​for pulmonary vascular resistance using conventional units are 0.25-1.6 mmHg min / l. Pulmonary vascular resistance can also be expressed in units of dynes / sec / cm5 (normal = 37-250 dynes / sec / cm5). One factor that contributes to increased PVR is hypoxemia. In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a peptide or peptides beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, The present invention relates to a method for reducing PVR in a patient by at least 10%, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0204] In some embodiments, the patient has a pulmonary vascular resistance (PVR) of 3 Wood units or more. In some embodiments, the method reduces the patient's PVR. In some embodiments, the method reduces the patient's PVR by at least 10%. In some embodiments, the method reduces the patient's PVR by at least 15%. In some embodiments, the method reduces the patient's PVR by at least 20%. In some embodiments, the method reduces the patient's PVR by at least 25%. In some embodiments, the method reduces the patient's PVR by at least 30%. In some embodiments, the method reduces the patient's PVR by at least 35%. In some embodiments, the method reduces the patient's PVR by at least 40%. In some embodiments, the method reduces the patient's PVR by at least 45%. In some embodiments, the method reduces the patient's PVR by at least 50%. In some embodiments, the method reduces the PVR to less than 3 Wood units.

[0205] DPG Pulmonary artery diastolic pressure gradient, DPG, has historically been used to determine the difference between diastolic pulmonary artery pressure and wedge pressure. In certain aspects, the present disclosure relates to a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a pulmonary hypertension inhibitor or pulmonary hypertension inhibitor starting at any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 290, 291, 292, 293, 294, 295, 296, 297, 298, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 300, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 32 The present invention relates to a method for lowering DPG in a patient by at least 10%, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0206] In some embodiments, the patient has a diastolic pressure gradient (DPG) of greater than 7 mmHg. In some embodiments, the patient has a DPG of at least 7 mmHg. In some embodiments, the patient has a DPG of at least 10 mmHg. In some embodiments, the patient has a DPG of at least 15 mmHg. In some embodiments, the patient has a DPG of at least 20 mmHg. In some embodiments, the patient has a DPG of at least 25 mmHg. In some embodiments, the patient has a DPG of at least 30 mmHg. In some embodiments, the patient has a DPG of at least 35 mmHg. In some embodiments, the patient has a DPG of at least 40 mmHg. In some embodiments, the patient has a DPG of at least 45 mmHg. In some embodiments, the patient has a DPG of at least 50 mmHg.

[0207] In some embodiments, the method reduces the patient's DPG. In some embodiments, the method reduces the patient's DPG by at least 10%. In some embodiments, the method reduces the patient's DPG by at least 15%. In some embodiments, the method reduces the patient's DPG by at least 20%. In some embodiments, the method reduces the patient's DPG by at least 25%. In some embodiments, the method reduces the patient's DPG by at least 30%. In some embodiments, the method reduces the patient's DPG by at least 35%. In some embodiments, the method reduces the patient's DPG by at least 40%. In some embodiments, the method reduces the patient's DPG by at least 45%. In some embodiments, the method reduces the patient's DPG by at least 50%. In some embodiments, the method lowers the patient's DPG to less than 7 mmHg.

[0208] BNP Brain natriuretic peptide (BNP) and NT-proBNP are markers of atrial and ventricular expansion due to elevated intracardiac pressure. The New York Heart Association (NYHA) has developed a four-stage functional classification system for congestive heart failure (CHF) based on the severity of symptoms. Studies have demonstrated that measured concentrations of circulating BNP and NT-proBNP increase with the severity of CHF based on the NYHA classification. In certain aspects, the present disclosure relates to a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a pulmonary hypertension drug or a combination thereof beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, The present invention relates to a method for lowering BNP levels in a patient, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of: 4, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0209] In some embodiments, the patient has an elevated BNP level compared to a healthy patient (e.g., a healthy individual of similar age and sex). In some embodiments, the patient has a normal BNP level. In some embodiments, the patient has a level of BNP of at least 100 pg / mL. In some embodiments, the patient has a level of BNP of at least 150 pg / mL. In some embodiments, the patient has a level of BNP of at least 200 pg / mL. In some embodiments, the patient has a level of BNP of at least 300 pg / mL. In some embodiments, the patient has a level of BNP of at least 400 pg / mL. In some embodiments, the patient has a level of BNP of at least 500 pg / mL. In some embodiments, the patient has a level of BNP of at least 1000 pg / mL. In some embodiments, the patient has a level of BNP of at least 3000 pg / mL. In some embodiments, the patient has a level of BNP of at least 5000 pg / mL. In some embodiments, the patient has a level of BNP of at least 10,000 pg / mL. In some embodiments, the patient has a level of BNP of at least 15,000 pg / mL. In some embodiments, the patient has a BNP level of at least 20,000 pg / mL.

[0210] In some embodiments, the method reduces the patient's BNP level by at least 10%. In some embodiments, the method reduces the patient's BNP level by at least 20%. In some embodiments, the method reduces the patient's BNP level by at least 25%. In some embodiments, the method reduces the patient's BNP level by at least 30%. In some embodiments, the method reduces the patient's BNP level by at least 35%. In some embodiments, the method reduces the patient's BNP level by at least 40%. In some embodiments, the method reduces the patient's BNP level by at least 45%. In some embodiments, the method reduces the patient's BNP level by at least 50%. In some embodiments, the method reduces the patient's BNP level by at least 55%. In some embodiments, the method reduces the patient's BNP level by at least 60%. In some embodiments, the method reduces the patient's BNP level by at least 65%. In some embodiments, the method reduces the patient's BNP level by at least 70%. In some embodiments, the method reduces the patient's BNP level by at least 75%. In some embodiments, the method reduces the patient's BNP level by at least 80%. In some embodiments, the methods reduce the patient's BNP levels to normal levels (ie, less than 100 pg / ml).

[0211] NT-proBNP In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a compound or medicament for use in treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, the ... The present invention relates to a method for lowering NT-proBNP levels in a patient, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 25, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0212] In some embodiments, the patient has an elevated NT-proBNP level compared to a healthy patient (e.g., a healthy individual of similar age and sex). In some embodiments, the patient has a normal NT-proBNP level. In some embodiments, the patient has an NT-proBNP level of at least 100 pg / mL. In some embodiments, the patient has an NT-proBNP level of at least 150 pg / mL. In some embodiments, the patient has an NT-proBNP level of at least 200 pg / mL. In some embodiments, the patient has an NT-proBNP level of at least 300 pg / mL. In some embodiments, the patient has an NT-proBNP level of at least 400 pg / mL. In some embodiments, the patient has an NT-proBNP level of at least 500 pg / mL. In some embodiments, the patient has an NT-proBNP level of at least 1000 pg / mL. In some embodiments, the patient has an NT-proBNP level of at least 3000 pg / mL. In some embodiments, the patient has an NT-proBNP level of at least 5000 pg / mL. In some embodiments, the patient has an NT-proBNP level of at least 10,000 pg / mL. In some embodiments, the patient has an NT-proBNP level of at least 15,000 pg / mL. In some embodiments, the patient has an NT-proBNP level of at least 20,000 pg / mL.

[0213] In some embodiments, the method reduces the patient's NT-proBNP level. In some embodiments, the method reduces the patient's NT-proBNP level by at least 10%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 20%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 25%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 30%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 35%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 40%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 45%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 50%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 55%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 60%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 65%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 70%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 75%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 80%. In some embodiments, the method reduces the patient's NT-proBNP level by at least 30%. In some embodiments, the method reduces the NT-proBNP level to normal levels. In some embodiments, normal levels of NT-proBNP are less than 100 pg / ml.

[0214] smooth muscle hypertrophy Patients with COPD frequently experience airway wall remodeling, primarily in small airways, resulting in airway wall thickening and airflow obstruction. Similarly, bronchial smooth muscle hypertrophy, characterized by an increase in smooth muscle cells and thickening of the smooth muscle layer surrounding the airways, is a hallmark of airway wall remodeling in disease states that resemble chronic asthma. In certain aspects, the present disclosure provides a method for treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a pulmonary disease, comprising administering to a patient in need thereof, a medicament for treating or preventing one or more complications of pulmonary hypertension associated with a pulmonary disease, the medicament comprising any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, and any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165 , 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135, comprising administering to the patient an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of

[0215] In some embodiments, the method reduces smooth muscle hypertrophy in the patient. In some embodiments, the method reduces smooth muscle hypertrophy in the patient by at least 10%. In some embodiments, the method reduces smooth muscle hypertrophy in the patient by at least 15%. In some embodiments, the method reduces smooth muscle hypertrophy in the patient by at least 20%. In some embodiments, the method reduces smooth muscle hypertrophy in the patient by at least 25%. In some embodiments, the method reduces smooth muscle hypertrophy in the patient by at least 30%. In some embodiments, the method reduces smooth muscle hypertrophy in the patient by at least 35%. In some embodiments, the method reduces smooth muscle hypertrophy in the patient by at least 40%. In some embodiments, the method reduces smooth muscle hypertrophy in the patient by at least 45%. In some embodiments, the method reduces smooth muscle hypertrophy in the patient by at least 50%.

[0216] Quality of life In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a therapeutically effective amount of a pulmonary hypertension inhibitor or pulmonary hypertension inhibitor, the therapeutically effective amount being selected from the group consisting of a pulmonary hypertension inhibitor or pulmonary hypertension inhibitor, ... The present invention relates to a method for improving the quality of life of a patient, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0217] In some embodiments, the patient's quality of life is measured using the Cambridge Pulmonary Hypertension Outcome Review (CAMPHOR). CAMPHOR is a disease-specific patient-reported outcome measure that assesses the quality of life of patients with pulmonary hypertension. Three dimensions of CAMPHOR assess symptoms, function, and quality of life. The Quality of Life (QoL) scale has 25 items focusing on socialization, role, recognition, self-esteem, independence, and safety. Similarly, the Symptoms dimension consists of 25 symptoms categorized into three subscales: vitality, shortness of breath, and mood. The Activity scale has 15 items. QoL and Symptom scores range from 0 to 25, with higher scores indicating poorer quality of life. Activity scores range from 0 to 30, with higher scores indicating more physical limitations. In some embodiments, the method reduces the patient's Quality of Life (QoL) score by at least 1%. In some embodiments, the method reduces the patient's Quality of Life (QoL) score by at least 2%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 3%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 4%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 5%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 10%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 15%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 20%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 25%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 30%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 35%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 40%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 45%.In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 50%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 55%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 60%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 65%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 70%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 75%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 80%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 85%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 90%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 95%. In some embodiments, the method reduces the patient's quality of life (QoL) score by at least 100%. In some embodiments, the patient's quality of life is improved as measured using the Cambridge Pulmonary Hypertension Outcome Review (CAMPHOR).

[0218] ventricular function In certain aspects, the disclosure relates to methods of improving or maintaining ventricular function (e.g., left ventricular function or right ventricular function). In some embodiments, the method improves right ventricular function in a patient. In some embodiments, the improvement in right ventricular function is due to an increase in right ventricular fractional area change. In some embodiments, the improvement in right ventricular function is due to a decrease in right ventricular hypertrophy. In some embodiments, the ejection fraction is improved. In some embodiments, the method improves right ventricular hypertrophy in a patient.

[0219] In certain aspects, the present disclosure relates to diagnostic tests and methods for pulmonary hypertension associated with lung disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD) or pulmonary fibrosis with emphysema (CPFE)). Echocardiography is a useful non-invasive screening tool for determining the severity of a patient's pulmonary hypertension. The improvement or maintenance of ventricular function (e.g., left ventricular function or right ventricular function) can be evaluated by a number of echocardiographic measurements. One such quantitative approach to assess ventricular function is the measurement of tricuspid annular systolic excursion (TAPSE). TAPSE estimates RV systolic function by measuring the level of systolic excursion of the lateral tricuspid annulus toward the cusp. Other echocardiographic measurements that may be used to assess maintenance and / or improvement of ventricular function include, but are not limited to, right ventricular fractional area change (RVFAC), right ventricular end-diastolic area (RVEDA), right ventricular end-systolic area (RVESA), right ventricular free wall thickness (RVFWT), right ventricular ejection fraction (RVEF), right ventricular-pulmonary artery (RV-PA) coupling, pulmonary artery systolic pressure (PASP), right ventricular systolic pressure (RVSP), pulmonary artery acceleration time (PAAT), tricuspid regurgitation velocity (TRV), left ventricular hypertrophy, and right ventricular hypertrophy.

[0220] TAPSE Tricuspid annular systolic excursion (TAPSE) can be obtained using echocardiography and represents a measure of RV longitudinal function. TAPSE has previously been shown to have a good correlation with parameters estimating RV global systolic function. A TAPSE of less than 17 mm strongly suggests RV systolic dysfunction. In some embodiments of the methods disclosed herein, the patient has a TAPSE of less than 20 mm. In some embodiments, the patient has a TAPSE of less than 18 mm. In some embodiments, the patient has a TAPSE of less than 16 mm. In some embodiments, the patient has a TAPSE of less than 14 mm. In some embodiments, the patient has a TAPSE of less than 12 mm.

[0221] In some embodiments, the method increases TAPSE to at least 20 mm. In some embodiments, the method increases TAPSE to at least 22 mm. In some embodiments, the method increases TAPSE to at least 24 mm. In some embodiments, the method increases TAPSE to at least 26 mm. In some embodiments, the method increases TAPSE to at least 28 mm. In some embodiments, the method increases TAPSE to at least 30 mm.

[0222] PASP and RVSP In certain aspects, the disclosure provides a method of treating pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a peptide comprising any one of the following amino acids beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191 The present invention relates to a method for reducing right ventricular systolic pressure (RVSP) by at least 10%, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 30, 131, 132, 133, 134, or 135.

[0223] In some embodiments, the PASP is resting PASP. In some aspects, the PASP is determined using tricuspid regurgitation velocity (TRV) and right aortic (RA) pressure. In some embodiments, the PASP is determined using the following formula:

[0224] PASP=TRV 2 ×4+RA pressure TRV has been shown to correlate with PASP at rest and during exercise. The pressure gradient between the right ventricle and the right atrium is calculated using the modified Bernoulli equation (Δp = 4V 2) can be used to calculate

[0225] In some embodiments, right ventricular systolic pressure (RVSP) is equal to PASP. In some embodiments, RVSP is measured in the absence of right ventricular outflow tract obstruction. In some embodiments, RVSP is determined using the following formula:

[0226] RVSP=4V 2 +RAP In the above equation, V represents the peak tricuspid regurgitant jet velocity and RAP is the mean right atrial pressure. RVSP is frequently used to estimate PASP.

[0227] In some embodiments, the patient has a right ventricular systolic pressure (RVSP) greater than 35 mmHg. In some embodiments, the method reduces the patient's RVSP. In some embodiments, the method reduces the patient's RVSP by at least 10%. In some embodiments, the method reduces the patient's RVSP by at least 15%. In some embodiments, the method reduces the patient's RVSP by at least 20%. In some embodiments, the method reduces the patient's RVSP by at least 25%. In some embodiments, the method reduces the patient's RVSP by at least 30%. In some embodiments, the method reduces the patient's RVSP by at least 35%. In some embodiments, the method reduces the patient's RVSP by at least 40%. In some embodiments, the method reduces the patient's RVSP by at least 45%. In some embodiments, the method reduces the patient's RVSP by at least 50%. In some embodiments, the method reduces the patient's RVSP to less than 25 mmHg.

[0228] In some embodiments, the patient has a pulmonary artery systolic pressure (PASP) greater than 20 mmHg. In some embodiments, the patient has a PASP greater than 25 mmHg. In some embodiments, the patient has a PASP of at least 35 mmHg. In some embodiments, the patient has a PASP of at least 40 mmHg. In some embodiments, the patient has a PASP of at least 50 mmHg. In some embodiments, the patient has a PASP of at least 55 mmHg. In some embodiments, the patient has a PASP of at least 60 mmHg.

[0229] In some embodiments, the method reduces the PASP of the patient. In some embodiments, the method reduces the PASP of the patient by at least 10%. In some embodiments, the method reduces the PASP of the patient by at least 15%. In some embodiments, the method reduces the PASP of the patient by at least 20%. In some embodiments, the method reduces the PASP of the patient by at least 25%. In some embodiments, the method reduces the PASP of the patient by at least 30%. In some embodiments, the method reduces the PASP of the patient by at least 35%. In some embodiments, the method reduces the PASP of the patient by at least 40%. In some embodiments, the method reduces the PASP of the patient by at least 45%. In some embodiments, the method reduces the PASP of the patient by at least 50%.

[0230] In some embodiments, the method reduces the patient's PASP by at least 5 mmHg. In some embodiments, the method reduces the patient's PASP by at least 10 mmHg. In some embodiments, the method reduces the patient's PASP by at least 15 mmHg. In some embodiments, the method reduces the patient's PASP by at least 20 mmHg. In some embodiments, the method reduces the patient's PASP by at least 25 mmHg. In some embodiments, the method reduces the patient's PASP to less than 25 mmHg. In some embodiments, the method reduces the patient's PASP to less than 20 mmHg.

[0231] right ventricular hypertrophy Right ventricular hypertrophy (RVH) is a pathological increase in muscle mass of the right ventricle in response to pressure overload, most commonly due to severe lung disease. Symptoms of RVH due to pulmonary hypertension include exertional chest pain, peripheral edema, exertional syncope, and right upper quadrant pain. In certain aspects, the present disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with lung disease, comprising administering to a patient in need thereof a pulmonary hypertension inhibitor or pulmonary hypertension inhibitor starting at any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 17 In one embodiment, the method comprises administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0232] In some embodiments, the method reduces right ventricular hypertrophy in the patient. In some embodiments, the method reduces right ventricular hypertrophy in the patient by at least 10%. In some embodiments, the method reduces right ventricular hypertrophy in the patient by at least 15%. In some embodiments, the method reduces right ventricular hypertrophy in the patient by at least 20%. In some embodiments, the method reduces right ventricular hypertrophy in the patient by at least 25%. In some embodiments, the method reduces right ventricular hypertrophy in the patient by at least 30%. In some embodiments, the method reduces right ventricular hypertrophy in the patient by at least 35%. In some embodiments, the method reduces right ventricular hypertrophy in the patient by at least 40%. In some embodiments, the method reduces right ventricular hypertrophy in the patient by at least 45%. In some embodiments, the method reduces right ventricular hypertrophy in the patient by at least 50%.

[0233] Cardiac index Cardiac index (CI) is an assessment of cardiac output based on a patient's size. Both cardiac output and cardiac index are important in determining whether the heart is pumping enough blood and delivering enough oxygen to cells. Cardiac index allows for comparison of cardiac function between individuals of different sizes. In certain aspects, the present disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with lung disease, comprising administering to a patient in need thereof a pulmonary hypertension inhibitor or pulmonary hypertension inhibitor starting at any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 1 The present invention relates to a method for increasing cardiac index, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of: 3, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0234] In some embodiments, the patient receives 2.5 L / min / m 2 In some embodiments, the patient has a cardiac index of less than 2.0 L / min / m 2 In some embodiments, the patient has a cardiac index of less than 1.5 L / min / m 2 In some embodiments, the patient has a cardiac index of less than 1.0 L / min / m 2In some embodiments, the method increases the patient's CI by at least 10%. In some embodiments, the method increases the patient's CI by at least 10%. In some embodiments, the method increases the patient's CI by at least 10%. In some embodiments, the method increases the patient's CI by at least 15%. In some embodiments, the method increases the patient's CI by at least 20%. In some embodiments, the method increases the patient's CI by at least 25%. In some embodiments, the method increases the patient's CI by at least 30%. In some embodiments, the method increases the patient's CI by at least 35%. In some embodiments, the method increases the patient's CI by at least 40%. In some embodiments, the method increases the patient's CI by at least 45%. In some embodiments, the method increases the patient's CI by at least 50%. In some embodiments, the method increases the patient's CI by at least 0.2 L / min / m 2 In some embodiments, the method increases the patient's CI by at least 0.4 L / min / m 2 In some embodiments, the method increases the patient's CI by at least 0.6 L / min / m 2 In some embodiments, the method increases the patient's CI by at least 0.8 L / min / m 2 In some embodiments, the method increases the patient's CI by at least 1 L / min / m 2 In some embodiments, the method increases the patient's CI by at least 1.2 L / min / m 2 In some embodiments, the method increases the patient's CI by at least 1.4 L / min / m 2 In some embodiments, the method increases the patient's CI by at least 1.6 L / min / m 2 In some embodiments, the method increases the patient's CI by at least 1.8 L / min / m 2 In some embodiments, the method increases the patient's CI by at least 2 L / min / m 2 In some embodiments, the method increases the patient's CI by at least 2.5 L / min / m 2 Increase to.

[0235] cardiac output Generally, normal cardiac output at rest is about 2.5-4.2 L / min / m2, and cardiac output may be decreased by approximately 40% without departing from the normal range. A low cardiac index below about 2.5 L / min / m2 generally indicates impaired cardiovascular function. Cardiac output can be used to calculate cardiac index (e.g., cardiac index = cardiac output / body surface area). Cardiac output can also be used to calculate stroke volume (e.g., stroke volume = CO / heart rate). In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a compound comprising a nucleotide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 34 , 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135, comprising administering to the patient an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of

[0236] In some embodiments, the patient's cardiac output is less than 4 L / min. In some embodiments, the method increases the patient's cardiac output by at least 10%. In some embodiments, the method increases the patient's cardiac output by at least 15%. In some embodiments, the method increases the patient's cardiac output by at least 20%. In some embodiments, the method increases the patient's cardiac output by at least 25%. In some embodiments, the method increases the patient's cardiac output by at least 30%. In some embodiments, the method increases the patient's cardiac output by at least 35%. In some embodiments, the method increases the patient's cardiac output by at least 40%. In some embodiments, the method increases the patient's cardiac output by at least 45%. In some embodiments, the method increases the patient's cardiac output by at least 50%. In some embodiments, the method increases the patient's cardiac output by at least 0.5 L / min. In some embodiments, the method increases the patient's cardiac output by at least 1 L / min. In some embodiments, the method increases the patient's cardiac output by at least 1.5 L / min. In some embodiments, the method increases the patient's cardiac output by at least 2 L / min. In some embodiments, the method increases the patient's cardiac output by at least 2.5 L / min. In some embodiments, the method increases the patient's cardiac output by at least 3 L / min. In some embodiments, the method increases the patient's cardiac output by at least 3.5 L / min. In some embodiments, the method increases the patient's cardiac output by at least 4 L / min.

[0237] Composite Physiological Index (CPI) The composite physiological index (CPI) can be used to determine the extent of pulmonary fibrosis. It is difficult to predict the clinical course of fibrotic lung diseases (e.g., idiopathic pulmonary fibrosis). The CPI model can be used as a predictor of fibrotic disease progression. In certain aspects, the present disclosure provides a method for treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with lung disease, comprising administering to a patient in need thereof a pulmonary hypertension inhibitor or pulmonary hypertension inhibitor starting at any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 290, 291, 292, 293, 294, 295, 296, 297, 298, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 300, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 32 The present invention relates to a method for reducing a composite physiological index, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 24, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0238] In some embodiments, the patient has a CPI greater than 15. In some embodiments of the methods herein, the patient has a CPI greater than 20. In some embodiments of the methods herein, the patient has a CPI greater than 25. In some embodiments of the methods herein, the patient has a CPI greater than 30. In some embodiments of the methods herein, the patient has a CPI greater than 35. In some embodiments of the methods herein, the patient has a CPI greater than 40. In some embodiments of the methods herein, the patient has a CPI greater than 45. In some embodiments of the methods herein, the patient has a CPI greater than 50. In some embodiments of the methods herein, the patient has a CPI greater than 55. In some embodiments of the methods herein, the patient has a CPI greater than 60. In some embodiments of the methods herein, the patient has a CPI greater than 65. In some embodiments of the methods herein, the patient has a CPI greater than 70. In some embodiments of the methods herein, the patient has a CPI greater than 75. In some embodiments of the methods herein, the patient has a CPI greater than 80. In some embodiments, the methods reduce the CPI of the patient. In some embodiments, the method reduces the patient's CPI by 10%. In some embodiments, the method reduces the patient's CPI by 15%. In some embodiments, the method reduces the patient's CPI by 20%. In some embodiments, the method reduces the patient's CPI by 25%. In some embodiments, the method reduces the patient's CPI by 30%. In some embodiments, the method reduces the patient's CPI by 35%. In some embodiments, the method reduces the patient's CPI by 40%. In some embodiments, the method reduces the patient's CPI by 45%. In some embodiments, the method reduces the patient's CPI by 50%. In some embodiments, the method reduces the CPI to less than 70. In some embodiments, the method reduces the CPI to less than 65. In some embodiments, the method reduces the CPI to less than 60. In some embodiments, the method reduces the CPI to less than 55. In some embodiments, the method reduces the CPI to less than 50.In some embodiments, the method reduces the CPI to less than 45. In some embodiments, the method reduces the CPI to less than 40. In some embodiments, the method reduces the CPI to less than 35. In some embodiments, the method reduces the CPI to less than 30. In some embodiments, the method reduces the CPI to less than 25. In some embodiments, the method reduces the CPI to less than 20. In some embodiments, the method reduces the CPI to less than 15. In some embodiments, the method reduces the CPI to less than 10. In some embodiments, the method reduces the CPI to less than 5.

[0239] Resting oxygen saturation In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a therapeutically effective amount of a pulmonary hypertension inhibitor or pulmonary hypertension inhibitor, the therapeutically effective amount being selected from the group consisting of a pulmonary hypertension inhibitor or pulmonary hypertension inhibitor, ... The present invention relates to a method for increasing arterial oxygen saturation, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0240] In some embodiments, the patient has an arterial oxygen saturation of less than 95%. In some embodiments of the methods disclosed herein, the patient has an arterial oxygen saturation of less than 90%. In some embodiments of the methods disclosed herein, the patient has an arterial oxygen saturation of less than 85%. In some embodiments of the methods disclosed herein, the patient has an arterial oxygen saturation of less than 80%. In some embodiments of the methods disclosed herein, the patient has an arterial oxygen saturation of less than 75%. In some embodiments of the methods disclosed herein, the patient has an arterial oxygen saturation of less than 70%. In some embodiments of the methods disclosed herein, the patient has an arterial oxygen saturation of less than 65%. In some embodiments of the methods disclosed herein, the patient has an arterial oxygen saturation of less than 60%. In some embodiments of the methods disclosed herein, the patient has an arterial oxygen saturation of less than 55%. In some embodiments of the methods disclosed herein, the patient has an arterial oxygen saturation of less than 50%. In some embodiments of the methods disclosed herein, the patient has an arterial oxygen saturation of less than 45%. In some embodiments of the methods disclosed herein, the patient's arterial oxygen saturation is less than 40%. In some embodiments of the methods disclosed herein, the patient's arterial oxygen saturation is less than 35%. In some embodiments of the methods disclosed herein, the patient's arterial oxygen saturation is less than 30%. In some embodiments, the method increases the patient's arterial oxygen saturation. In some embodiments, the method increases the patient's arterial oxygen saturation by at least 5%.

[0241] In some embodiments, the method increases the patient's arterial oxygen saturation by at least 10%. In some embodiments, the method increases the patient's arterial oxygen saturation by at least 15%. In some embodiments, the method increases the patient's arterial oxygen saturation by at least 20%. In some embodiments, the method increases the patient's arterial oxygen saturation by at least 25%. In some embodiments, the method increases the patient's arterial oxygen saturation by at least 30%. In some embodiments, the method increases the patient's arterial oxygen saturation by at least 35%. In some embodiments, the method increases the patient's arterial oxygen saturation by at least 40%. In some embodiments, the method increases the patient's arterial oxygen saturation by at least 45%. In some embodiments, the method increases the patient's arterial oxygen saturation by at least 50%. In some embodiments, the method increases the patient's arterial oxygen saturation by at least 85%. In some embodiments, the method increases the patient's arterial oxygen saturation by at least 90%. In some embodiments, the method increases the patient's arterial oxygen saturation by at least 95%. In some embodiments, the arterial oxygen saturation is measured at rest.

[0242] Exercise capacity (6MWD and BDI) In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a compound comprising any one of the following amino acids beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, The present invention relates to a method of increasing exercise tolerance in a patient, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 24, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0243] Any suitable measure of exercise tolerance can be used. For example, exercise tolerance in the 6-minute walk test (6MWT), which measures the distance a subject can walk in 6 minutes, i.e., 6-minute walk distance (6MWD), is frequently used to assess the severity and disease progression of pulmonary hypertension. The BDI is a numerical scale for assessing perceived dyspnea (discomfort in breathing) and can be used to measure exercise tolerance. It measures the degree of shortness of breath, for example, after completing the 6MWT, with a BDI of 0 indicating no shortness of breath and 10 indicating maximum shortness of breath. In some embodiments, the patient has a 6-minute walk distance (6MWD) of less than 550 meters before treatment. In some embodiments, the patient has a 6-minute walk distance (6MWD) of less than 550 meters before treatment. In some embodiments, the patient has a 6-minute walk distance (6MWD) of less than 500 meters before treatment. In some embodiments, the patient has a 6-minute walk distance (6MWD) of less than 450 meters before treatment. In some embodiments, the patient has a 6-minute walk distance (6MWD) of less than 400 meters before treatment. In some embodiments, the patient has a 6 minute walk distance (6MWD) of less than 350 meters prior to treatment. In some embodiments, the patient has a 6 minute walk distance (6MWD) of less than 300 meters prior to treatment. In some embodiments, the patient has a 6 minute walk distance (6MWD) of less than 250 meters prior to treatment. In some embodiments, the patient has a 6 minute walk distance (6MWD) of less than 200 meters prior to treatment. In some embodiments, the patient has a 6 minute walk distance (6MWD) of less than 150 meters prior to treatment. In some embodiments, the method increases the patient's 6MWD by at least 10 meters. In some embodiments, the method increases the patient's 6MWD by at least 15 meters. In some embodiments, the method increases the patient's 6MWD by at least 20 meters. In some embodiments, the method increases the patient's 6MWD by at least 25 meters. In some embodiments, the method increases the patient's 6MWD by at least 30 meters. In some embodiments, the method increases the patient's 6MWD by at least 35 meters.In some embodiments, the method increases the 6MWD of the patient by at least 40 meters. In some embodiments, the method increases the 6MWD of the patient by at least 45 meters. In some embodiments, the method increases the 6MWD of the patient by at least 50 meters. In some embodiments, the method increases the 6MWD of the patient by at least 55 meters. In some embodiments, the method increases the 6MWD of the patient by at least 60 meters. In some embodiments, the method increases the 6MWD of the patient by at least 65 meters. In some embodiments, the method increases the 6MWD of the patient by at least 70 meters. In some embodiments, the method increases the 6MWD of the patient by at least 75 meters. In some embodiments, the method increases the 6MWD of the patient by at least 80 meters. In some embodiments, the method increases the 6MWD of the patient by at least 85 meters. In some embodiments, the method increases the 6MWD of the patient by at least 90 meters. In some embodiments, the method increases the 6MWD of the patient by at least 95 meters. In some embodiments, the method increases the 6MWD of the patient by at least 100 meters. In some embodiments, the method increases the 6MWD of the patient by at least 125 meters. In some embodiments, the method increases the patient's 6MWD by at least 150 meters. In some embodiments, the method increases the patient's 6MWD by at least 175 meters. In some embodiments, the method increases the patient's 6MWD by at least 200 meters. In some embodiments, the method increases the patient's 6MWD by at least 250 meters. In some embodiments, the method increases the patient's 6MWD by at least 300 meters. In some embodiments, the method increases the patient's 6MWD by at least 400 meters.

[0244] In some embodiments, the method increases the patient's exercise tolerance. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 0.5 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 1 index point. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 1.5 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 2 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 2.5 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 3 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 3.5 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 4 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 4.5 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 5 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 5.5 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 6 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 6.5 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 7 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 7.5 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 8 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 8.5 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 9 index points. In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 9.5 index points.In some embodiments, the patient has a Borg Dyspnea Index (BDI) of at least 10 index points. In some embodiments, the method reduces the patient's Borg Dyspnea Index (BDI). In some embodiments, the method reduces the patient's BDI by at least 0.5 index points. In some embodiments, the method reduces the patient's BDI by at least 1 index point. In some embodiments, the method reduces the patient's BDI by at least 1.5 index points. In some embodiments, the method reduces the patient's BDI by at least 2 index points. In some embodiments, the method reduces the patient's BDI by at least 2.5 index points. In some embodiments, the method reduces the patient's BDI by at least 3 index points. In some embodiments, the method reduces the patient's BDI by at least 3.5 index points. In some embodiments, the method reduces the patient's BDI by at least 4 index points. In some embodiments, the method reduces the patient's BDI by at least 4.5 index points. In some embodiments, the method reduces the patient's BDI by at least 5 index points. In some embodiments, the method reduces the patient's BDI by at least 5.5 index points. In some embodiments, the method reduces the patient's BDI by at least 6 index points. In some embodiments, the method reduces the patient's BDI by at least 6.5 index points. In some embodiments, the method reduces the patient's BDI by at least 7 index points. In some embodiments, the method reduces the patient's BDI by at least 7.5 index points. In some embodiments, the method reduces the patient's BDI by at least 8 index points. In some embodiments, the method reduces the patient's BDI by at least 8.5 index points. In some embodiments, the method reduces the patient's BDI by at least 9 index points. In some embodiments, the method reduces the patient's BDI by at least 9.5 index points. In some embodiments, the method reduces the patient's BDI by at least 10 index points.

[0245] Pulmonary function tests In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a compound comprising a nucleotide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 34 , 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135, comprising administering to a patient an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of

[0246] Spirometry, or breathing measurement, is the primary pulmonary function test that can determine the volume and / or rate (flow rate) of air a subject inhales and exhales. Spirometers are used to measure forced vital capacity (FVC) (measured in liters and / or percentage of predicted) in forced expiratory volume (FEV) tests, among other characteristics. In an FEV test, a subject inhales deeply and exhales into a sensor for as long as possible (e.g., at least 6 seconds). Spirometry can also be used to test inhalation. FEV tests are usually repeated at least three times to ensure accuracy. The "normal" range for FVC is typically considered to be 80%-100% of predicted. "Of predicted" refers to reporting a subject's results as a percentage of a known predicted value for healthy subjects with similar characteristics (e.g., height, sex, age, race, weight). Other measurements that can be obtained include, but are not limited to, FEV1, where FVC is measured within the first second of a forced exhalation, and / or forced expiratory flow (FEF), which measures the flow of air leaving the lungs during the mid-portion of a forced exhalation. FEV1 / FVC ratios are also typically calculated.

[0247] In some embodiments of the methods disclosed herein, the patient has a forced vital capacity in one second (FEV1) of greater than 70%. In some embodiments of the methods disclosed herein, the patient has a forced vital capacity in one second (FEV1) of 60%-69%. In some embodiments of the methods disclosed herein, the patient has a forced vital capacity in one second (FEV1) of 50%-59%. In some embodiments of the methods disclosed herein, the patient has a forced vital capacity in one second (FEV1) of 35%-49%. In some embodiments of the methods disclosed herein, the patient has a forced vital capacity in one second (FEV1) of less than 35%. In some embodiments, the method increases the patient's FEV1. In some embodiments, the method increases the patient's FEV1 by at least 5%. In some embodiments, the method increases the patient's FEV1 by at least 10%. In some embodiments, the method increases the patient's FEV1 by at least 15%. In some embodiments, the method increases the patient's FEV1 by at least 20%. In some embodiments, the method increases the patient's FEV1 by at least 25%. In some embodiments, the method increases the patient's FEV1 by at least 30%. In some embodiments, the method increases the patient's FEV1 by at least 35%. In some embodiments, the method increases the patient's FEV1 by at least 40%. In some embodiments, the method increases the patient's FEV1 by at least 45%. In some embodiments, the method increases the patient's FEV1 by at least 50%. In some embodiments, the method increases the FEV1 by at least 60%. In some embodiments, the method increases the FEV1 by at least 65%. In some embodiments, the method increases the FEV1 by at least 70%. In some embodiments, the method increases the FEV1 by at least 75%. In some embodiments, the method increases the FEV1 by at least 80%. In some embodiments, the method increases the FEV1 by at least 85%. In some embodiments, the method increases the FEV1 by at least 90%. In some embodiments, the method increases the FEV1 by at least 95%.

[0248] In some embodiments, the patient has a forced vital capacity (FVC) of greater than 80%. In some embodiments, the patient has a forced vital capacity (FVC) of greater than 70%.

[0249] In some embodiments, the patient has a forced vital capacity (FVC) of 60%-69%. In some embodiments, the patient has a forced vital capacity (FVC) of 50%-59%. In some embodiments, the patient has a forced vital capacity (FVC) of 35%-49%. In some embodiments, the patient has a forced vital capacity (FVC) of less than 35%.

[0250] In some embodiments, the method increases the patient's FVC. In some embodiments, the method increases the patient's FVC by at least 5%. In some embodiments, the method increases the patient's FVC by at least 10%. In some embodiments, the method increases the patient's FVC by at least 15%. In some embodiments, the method increases the patient's FVC by at least 20%. In some embodiments, the method increases the patient's FVC by at least 25%. In some embodiments, the method increases the patient's FVC by at least 30%. In some embodiments, the method increases the patient's FVC by at least 35%. In some embodiments, the method increases the patient's FVC by at least 40%. In some embodiments, the method increases the patient's FVC by at least 45%. In some embodiments, the method increases the patient's FVC by at least 50%. In some embodiments, the method increases the FVC by at least 60%. In some embodiments, the method increases the FVC by at least 65%. In some embodiments, the method increases the FVC by at least 70%. In some embodiments, the method increases the FVC by at least 75%. In some embodiments, the method increases the FVC by at least 80%. In some embodiments, the method increases the FVC by at least 85%. In some embodiments, the method increases the FVC by at least 90%. In some embodiments, the method increases the FVC by at least 95%.

[0251] Carbon monoxide transfer coefficient Kco In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a compound comprising a nucleotide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 10 The present invention relates to a method for increasing carbon monoxide diffusing capacity in a patient, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of: 4, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0252] Carbon monoxide diffusing capacity or DL CO can be used in conjunction with spirometry and lung volume assessment to diagnose underlying lung disease (e.g., DL CO (Normal spirometry and lung volumes associated with decreased CO2 may suggest anemia, pulmonary vascular disease, early ILD, or early emphysema.) In some embodiments, the patient has a carbon monoxide diffusing capacity (DL) of less than 60%. CO In some embodiments, the patient has a carbon monoxide diffusing capacity (DL) of less than 55%. CO In some embodiments, the patient has a carbon monoxide diffusing capacity (DL) of less than 50%. CO In some embodiments, the patient has a carbon monoxide diffusing capacity (DL) of less than 45%. CO In some embodiments, the patient has a carbon monoxide diffusing capacity (DL) of less than 40%. CO In some embodiments, the patient has a carbon monoxide diffusing capacity (DL) of less than 35%. CO In some embodiments, the patient has a carbon monoxide diffusing capacity (DL) of less than 30%. CO In some embodiments, the patient has a carbon monoxide diffusing capacity (DL) of less than 25%.CO In some embodiments, the patient has a carbon monoxide diffusing capacity (DL) of less than 20%. CO ).

[0253] In some embodiments, the method further comprises administering to the patient a DL CO In some embodiments, the method increases the DL CO In some embodiments, the method increases the DL of the patient by at least 5%. CO In some embodiments, the method increases the DL of the patient by at least 10%. CO In some embodiments, the method increases the DL of the patient by at least 15%. CO In some embodiments, the method increases the DL of the patient by at least 20%. CO In some embodiments, the method increases the DL of the patient by at least 25%. CO In some embodiments, the method increases the DL of the patient by at least 30%. CO In some embodiments, the method increases the DL of the patient by at least 35%. CO In some embodiments, the method increases the DL of the patient by at least 40%. CO In some embodiments, the method increases the patient's DL by at least 45%. CO In some embodiments, the method increases DL CO In some embodiments, the method increases DL CO In some embodiments, the method increases DL CO In some embodiments, the method increases DL CO In some embodiments, the method increases DL CO In some embodiments, the method increases DL CO Increase by at least 65%.

[0254] Pulmonary fibrosis In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a therapeutically effective amount of a pulmonary hypertension inhibitor or pulmonary hypertension inhibitor, the therapeutically effective amount being selected from the group consisting of a pulmonary hypertension inhibitor or pulmonary hypertension inhibitor, ... The present invention relates to a method of reducing pulmonary fibrosis in a patient, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0255] In some embodiments, the method reduces pulmonary fibrosis in the patient by at least 10%. In some embodiments, the method reduces pulmonary fibrosis in the patient by at least 15%. In some embodiments, the method reduces pulmonary fibrosis in the patient by at least 20%. In some embodiments, the method reduces pulmonary fibrosis in the patient by at least 25%. In some embodiments, the method reduces pulmonary fibrosis in the patient by at least 30%. In some embodiments, the method reduces pulmonary fibrosis in the patient by at least 35%. In some embodiments, the method reduces pulmonary fibrosis in the patient by at least 40%. In some embodiments, the method reduces pulmonary fibrosis in the patient by at least 45%. In some embodiments, the method reduces pulmonary fibrosis in the patient by at least 50%.

[0256] Transplant-free survival rate In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a compound comprising any one of the following amino acids beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1, including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, The present invention relates to a method for increasing transplant-free survival in a patient, comprising administering an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of 24, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135.

[0257] In some embodiments, the method increases the patient's transplant-free survival by at least 10%. In some embodiments, the method increases the patient's transplant-free survival by at least 15%. In some embodiments, the method increases the patient's transplant-free survival by at least 20%. In some embodiments, the method increases the patient's transplant-free survival by at least 25%. In some embodiments, the method increases the patient's transplant-free survival by at least 30%. In some embodiments, the method increases the patient's transplant-free survival by at least 35%. In some embodiments, the method increases the patient's transplant-free survival by at least 40%. In some embodiments, the method increases the patient's transplant-free survival by at least 45%. In some embodiments, the method increases the patient's transplant-free survival by at least 50%.

[0258] death In certain aspects, the disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease, comprising administering to a patient in need thereof a compound comprising a nucleotide sequence beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 34 , 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135, comprising administering to the patient an effective amount of a polypeptide comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence ending in any one of

[0259] In some embodiments, the method reduces the risk of death associated with pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)) by at least 10%. In some embodiments, the method reduces the risk of death associated with pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)) by at least 15%. In some embodiments, the method reduces the risk of death associated with pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)) by at least 20%. In some embodiments, the method reduces the risk of death associated with pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)) by at least 25%. In some embodiments, the method reduces the risk of death associated with pulmonary hypertension associated with pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)) by at least 30%. In some embodiments, the method reduces the risk of death associated with pulmonary hypertension associated with pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)) by at least 35%. In some embodiments, the method reduces the risk of death associated with pulmonary hypertension associated with pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)) by at least 40%. In some embodiments, the method reduces the risk of death associated with pulmonary hypertension associated with pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)) by at least 45%.In some embodiments, the method reduces the risk of death associated with pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or combined pulmonary fibrosis and emphysema (CPFE)) by at least 50%.

[0260] Combination therapy In some cases, the disclosed methods for treating, preventing, or reducing the rate of progression and / or severity of pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pre-emphysema pulmonary fibrosis (CPFE)), in particular for treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pre-emphysema pulmonary fibrosis (CPFE)), may further comprise administering to the patient one or more supportive therapies or additional active agents for treating pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pre-emphysema pulmonary fibrosis (CPFE)). For example, patients may also be prescribed medications such as nitrates, hydralazines, pyridones (e.g., pirfenidone), small molecule tyrosine-kinase inhibitors (e.g., nintedanib), prostacyclin and its derivatives (e.g., epoprostenol, treprostinil, and iloprost); prostacyclin receptor agonists (e.g., selexipag); endothelin receptor antagonists (e.g., thelin, ambrisentan, macitentan, darusentan, and bosentan); calcium channel blockers (e.g., amlodipine, diltiazem, and nifedipine); anticoagulants (e.g., warfarin); diuretics; oxygen therapy; atrial septotomy; pulmonary endarterectomy; phosphodiesterase type 5 inhibitors (e.g., sildenafil and tadalafil). fil); activators of soluble guanylate cyclase (e.g., cinaciguat, veruiciguat and riociguat); ASK-1 inhibitors (e.g., CIIA; SCH79797; GS-4997; MSC2032964A; 3H-naphtho[1,2,3-de]quinyline-2,7-dione, NQDI-1; 2-thioxo-thiazolidine, 5-bromo-3-(4-oxo-2-thioxo-thiazolidine-5-ylidene)-1,3-dihydro-indol-2-one); NF-κB antagonists (e.g., dh404, CDDO-epoxide; 2.2-difluoropropionamide; C28 imidazole (CDDO-Im); 2-cyano-3,12-dioxooleane-1,9-dien-28-oic acid (CDDO);3-Acetyloleanolic acid;3-Trifluoroacetyloleanolic acid;28-Methyl-3-acetyloleanan;28-Methyl-3-trifluoroacetyloleanan;28-Methyloxyoleanolic acid;SZC014;SCZ015;SZC017;PEGylated derivatives of oleanolic acid;3-O-(β-D-glucopyranosyl)oleanolic acid;3-O-[β-D-glucopyranosyl-(1-- 3-O-[β-D-glucopyranosyl-(1-->2)-β-D-glucopyranosyl]oleanolic acid;3-O-[β-D-glucopyranosyl-(1-->3)-β-D-glucopyranosyl]oleanolic acid 28-O-β-D-glucopyranosyl ester;3-O-[β-D-glucopyranosyl-(1-->2 ... oleanolic acid 28-O-β-D-glucopyranosyl ester;3-O-[α-L-rhamnopyranosyl-(1-->3)-β-D-glucuronopyranosyl]oleanolic acid;3-O-[α-L-rhamnopyranosyl-(1-->3)-β-D-glucuronopyranosyl]oleanolic acid 28-O-β-D-glucopyranosyl ester;28-O-β-D-Glucopyranosyl-oleanolic acid;3-O-β-D- Glucopyranosyl(1→3)-β-D-glucopyranosiduronic acid (CS1);Oleanolic acid 3-O-β-D-glucopyranosyl(1→3)-β-D-glucopyranosiduronic acid (CS2);Methyl 3,11-dioxoolean-12-en-28-oleate (DIOXOL);ZCVI4-2;Benzyl 3-dehydro-oxy-1,2,5-oxadiazolo[3',4':2,3]oleanolate);The patient may be administered one or more supportive therapies or active agents selected from the group consisting of: , oxygen therapy, lung and / or heart transplant. In some embodiments, the methods described herein may further comprise administering pirfenidone to the patient. In some embodiments, the methods described herein may further comprise administering nintedanib to the patient. In some embodiments, the methods described herein may further comprise administering oral prostacyclin to the patient. In some embodiments, the methods described herein may further comprise administering one additional supportive therapy or additional active agent (i.e., dual therapy) to the patient for treating pulmonary hypertension associated with pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)). In some embodiments, the methods described herein may further include administering to the patient two additional supportive therapies or additional active agents (i.e., triple therapy) for treating pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)). In some embodiments, the methods described herein may further include administering to the patient three additional supportive therapies or additional active agents (i.e., quadruple therapy) for treating pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)).

[0261] In some embodiments, the methods described herein may further comprise administering to the patient an angiotensin antagonist (e.g., angiotensin receptor blocker, ARB). In some embodiments, the patient is further administered one or more ARBs selected from the group consisting of losartan, irbesartan, olmesartan, candesartan, valsartan, fimasartan, azilsartan, salplisartan, and telmisartan. In some embodiments, the patient is administered losartan. In some embodiments, the patient is administered irbesartan. In some embodiments, the patient is administered olmesartan. In some embodiments, the patient is administered candesartan. In some embodiments, the patient is administered valsartan. In some embodiments, the patient is administered fimasartan. In some embodiments, the patient is administered azilsartan. In some embodiments, the patient is administered salplisartan. In some embodiments, the patient is administered telmisartan.

[0262] In some embodiments, the methods described herein may further comprise administering to the patient one or more ACE inhibitors. In some embodiments, the one or more ACE inhibitors are selected from the group consisting of benazepril, captopril, enalapril, lisinopril, perindopril, ramipril (e.g., ramipen), trandolapril, and zofenopril. In some embodiments, the patient is administered benazepril. In some embodiments, the patient is administered captopril. In some embodiments, the patient is administered enalapril. In some embodiments, the patient is administered lisinopril. In some embodiments, the patient is administered perindopril. In some embodiments, the patient is administered ramipril. In some embodiments, the patient is administered trandolapril. In some embodiments, the patient is administered zofenopril. In some embodiments, the methods described herein may further comprise administering to the patient an ARB and an ACE inhibitor. In some embodiments, an alternative approach to angiotensin antagonism is to combine an ACE inhibitor and / or an ARB with an aldosterone antagonist.

[0263] In some embodiments, one or more supportive therapies or additional active agents for treating pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)) are administered prior to administration of an ActRII polypeptide. In some embodiments, one or more supportive therapies or additional active agents for treating pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)) are administered in combination with an ActRII polypeptide. In some embodiments, one or more supportive therapies or additional active agents for treating pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary fibrosis with emphysema (CPFE)) are administered after administration of an ActRII polypeptide.

[0264] Functional classification Pulmonary hypertension associated with lung disease at baseline (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD) or pulmonary fibrosis with emphysema (CPFE)) can be mild, moderate or severe, as measured, for example, by the World Health Organization (WHO) functional classification, which is a measure of disease severity in patients with pulmonary hypertension. The WHO functional classification is an adaptation of the New York Heart Association (NYHA) system and is routinely used, for example, in monitoring disease progression and response to treatment, to qualitatively assess activity tolerance (Rubin (2004) Chest 126:7-10). The WHO system recognizes four functional classes: functional class I: pulmonary hypertension that does not limit physical activity; ordinary physical activity does not lead to excessive dyspnea or fatigue, chest pain or presyncope; functional class II: pulmonary hypertension that leads to slight limitation of physical activity; the patient is comfortable at rest; ordinary physical activity leads to excessive dyspnea or fatigue, chest pain or presyncope; functional class III: pulmonary hypertension that leads to significant limitation of physical activity; the patient is comfortable at rest; less than ordinary activity leads to excessive dyspnea or fatigue, chest pain or presyncope; functional class IV: pulmonary hypertension that leads to symptoms with any physical activity; the patient shows signs of right heart failure; dyspnea and / or fatigue may be present even at rest; any physical activity leads to increased discomfort. In some embodiments of the methods disclosed herein, the methods prevent or reduce the progression of pulmonary hypertension functional classes recognized by the World Health Organization (WHO). In some embodiments, the method prevents or reduces the progression of pulmonary hypertension functional class I to class II pulmonary hypertension as recognized by the WHO. In some embodiments, the method prevents or reduces the progression of pulmonary hypertension functional class II to class III pulmonary hypertension as recognized by the WHO. In some embodiments, the method prevents or reduces the progression of pulmonary hypertension functional class III to class IV pulmonary hypertension as recognized by the WHO. In some embodiments, the method promotes or enhances the regression of pulmonary hypertension functional class as recognized by the WHO.In some embodiments, the method promotes or enhances regression of WHO-recognized pulmonary hypertension functional classification from Class IV to Class III pulmonary hypertension. In some embodiments, the method promotes or enhances regression of WHO-recognized pulmonary hypertension functional classification from Class III to Class II pulmonary hypertension. In some embodiments, the method promotes or enhances regression of WHO-recognized pulmonary hypertension functional classification from Class II to Class I pulmonary hypertension.

[0265] In some embodiments, the disclosure relates to a method of preventing or reducing the progression of a pulmonary hypertension functional class, comprising administering to a patient in need thereof an effective amount of an ActRII polypeptide (e.g., an amino acid sequence at least 90% identical to an amino acid sequence corresponding to residues 30-110 of SEQ ID NO:1). In some embodiments, the reduction in functional class progression is a delay in functional class progression. In some embodiments, the method relates to preventing or reducing the progression of a pulmonary hypertension functional class as recognized by the WHO. In some embodiments, the method relates to a patient having functional class I pulmonary hypertension as recognized by the WHO. In some embodiments, the method relates to preventing or reducing the progression of a patient from functional class I pulmonary hypertension to functional class II pulmonary hypertension as recognized by the WHO. In some embodiments, the method relates to a patient having functional class II pulmonary hypertension as recognized by the WHO. In some embodiments, the method relates to preventing or reducing the progression of a patient from functional class II pulmonary hypertension to functional class III pulmonary hypertension as recognized by the WHO. In some embodiments, the method relates to a patient having functional class III pulmonary hypertension as recognized by the WHO. In some embodiments, the methods relate to preventing or reducing the progression of a patient from functional class III pulmonary hypertension to functional class IV pulmonary hypertension as recognized by the WHO.

[0266] In certain aspects, the disclosure provides a method of promoting or enhancing regression of pulmonary hypertension functional class in pulmonary hypertension associated with a pulmonary disease (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary hypertension associated with combined emphysema and pulmonary fibrosis (CPFE)), comprising administering to a patient in need thereof an agonist or antagonist, comprising administering to said patient an agonist or antagonist, comprising administering to said patient an agonist or antagonist, comprising administering to said patient an agonist or antagonist, comprising administering to said patient an agonist or antagonist, comprising administering to said patient an agonist or antagonist, comprising administering to said patient an agonist or antagonist, In some embodiments, the patient has functional class I, functional class II, functional class III or functional class IV pulmonary hypertension as recognized by the WHO. In some embodiments, the method relates to a patient having functional class II pulmonary hypertension as recognized by the WHO. In some embodiments, the method relates to promoting regression of a patient from functional class II pulmonary hypertension to functional class I pulmonary hypertension as recognized by the WHO. In some embodiments, the method relates to a patient having functional class III pulmonary hypertension as recognized by the WHO. In some embodiments, the method relates to promoting regression of a patient from functional class III pulmonary hypertension to functional class II pulmonary hypertension as recognized by the WHO. In some embodiments, the method relates to promoting regression of a patient from functional class III pulmonary hypertension to functional class I pulmonary hypertension as recognized by the WHO. In some embodiments, the method relates to a patient having functional class IV pulmonary hypertension as recognized by the WHO. In some embodiments, the method relates to promoting regression of a patient from functional class IV pulmonary hypertension to functional class III pulmonary hypertension as recognized by the WHO.In some embodiments, the methods relate to promoting regression of a patient from functional class IV pulmonary hypertension to functional class II pulmonary hypertension as recognized by the WHO. In some embodiments, the methods relate to promoting regression of a patient from functional class IV pulmonary hypertension to functional class I pulmonary hypertension as recognized by the WHO.

[0267] In some embodiments, the regression of functional class is examined after the patient has undergone treatment utilizing an ActRII polypeptide disclosed herein for 4 weeks. In some embodiments, the regression of functional class is examined after the patient has undergone treatment utilizing an ActRII polypeptide disclosed herein for 8 weeks. In some embodiments, the regression of functional class is examined after the patient has undergone treatment utilizing an ActRII polypeptide disclosed herein for 12 weeks. In some embodiments, the regression of functional class is examined after the patient has undergone treatment utilizing an ActRII polypeptide disclosed herein for 16 weeks. In some embodiments, the regression of functional class is examined after the patient has undergone treatment utilizing an ActRII polypeptide disclosed herein for 20 weeks. In some embodiments, the regression of functional class is examined after the patient has undergone treatment utilizing an ActRII polypeptide disclosed herein for 22 weeks. In some embodiments, the regression of functional class is examined after the patient has undergone treatment utilizing an ActRII polypeptide disclosed herein for 24 weeks. In some embodiments, the regression of functional class is examined after the patient has undergone 26 weeks of treatment utilizing an ActRII polypeptide disclosed herein. In some embodiments, the regression of functional class is examined after the patient has undergone 28 weeks of treatment utilizing an ActRII polypeptide disclosed herein. In some embodiments, the regression of functional class is examined after the patient has undergone 48 weeks of treatment utilizing an ActRII polypeptide disclosed herein.

[0268] Long-lasting therapeutic effect In certain aspects, the present disclosure provides a method of treating, preventing, or reducing the rate of progression and / or severity of one or more complications of pulmonary hypertension associated with a lung disease in a sustained manner, comprising administering to a patient in need thereof a therapeutically effective amount of a compound or a combination thereof beginning with any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO:1 and including any one of amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 1 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135. In some embodiments, the sustained manner comprises a sustained therapeutic effect after reducing administration of an ActRII polypeptide described herein. In some embodiments, the sustained manner comprises a sustained therapeutic effect after ceasing administration of an ActRII polypeptide described herein. In some embodiments, the sustained therapeutic effect comprises maintaining a functional or hematological measure over time. In some embodiments, a sustained therapeutic effect is measured as a sustained reduction in PVR. In some embodiments, the patient's PVR level does not increase for at least 1 week to at least 12 weeks after cessation of the ActRII polypeptide treatment described herein. In some embodiments, the patient's PVR level does not increase for at least 1 week after cessation of the ActRII polypeptide treatment described herein. In some embodiments, the patient's PVR level does not increase for at least 2 weeks after cessation of the ActRII polypeptide treatment described herein. In some embodiments, the patient's PVR level does not increase for at least 3 weeks after cessation of the ActRII polypeptide treatment described herein. In some embodiments, the patient's PVR level does not increase for at least 4 weeks after cessation of the ActRII polypeptide treatment described herein.In some embodiments, the patient's PVR level does not increase for at least 5 weeks after cessation of the ActRII polypeptide treatment described herein. In some embodiments, the patient's PVR level does not increase for at least 6 weeks after cessation of the ActRII polypeptide treatment described herein. In some embodiments, the patient's PVR level does not increase for at least 1 month to at least 6 months after cessation of the ActRII polypeptide treatment described herein.

[0269] In certain aspects, the disclosure relates to a method of treating or preventing cardiopulmonary remodeling associated with pulmonary hypertension associated with a pulmonary disease in a patient (e.g., pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary hypertension associated with combined emphysema and pulmonary fibrosis (CPFE)), comprising administering to a patient in need thereof an effective amount of an ActRIIA polypeptide, said method slowing down cardiac remodeling and / or reversing cardiac remodeling. In some embodiments, the reversal is sustained reversal. In some embodiments, the cardiac remodeling is ventricular remodeling. In some embodiments, the ventricular remodeling is left ventricular remodeling. In some embodiments, the ventricular remodeling is right ventricular remodeling. In some embodiments, the cardiac remodeling is ventricular dilation. In some embodiments, the method reduces end diastole of the interventricular septum. In some embodiments, the method reduces end diastole of the posterior ventricular septum.

[0270] In some embodiments, echocardiographic measurements can be used to assess sustained therapeutic effects. In some embodiments, echocardiographic measurements include, but are not limited to, RV fractional area change (RVFAC), sPAP, tricuspid annular systolic velocity (TASV), and Tei index. In some embodiments, patients treated with the ActRIIA polypeptides disclosed herein exhibit sustained therapeutic effects. In some embodiments, sustained therapeutic effects result in a reduction in abdominal wall intrusion into the left ventricle. In some embodiments, sustained therapeutic effects result in an increase in right ventricular fractional area change (RVFAC).

[0271] Measurement of various parameters over time In certain embodiments, one or more of the pulmonary hypertension measurements described herein (e.g., pulmonary hypertension associated with a lung disease (e.g., chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), or pulmonary hypertension associated with emphysema combined with pulmonary fibrosis (CPFE))) can be measured over various treatment periods. In some embodiments, one or more of the pulmonary hypertension measurements described herein are measured after the patient has been treated with an ActRII polypeptide disclosed herein for 4 weeks. In some embodiments, one or more of the pulmonary hypertension measurements described herein are measured after the patient has been treated with an ActRII polypeptide disclosed herein for 8 weeks. In some embodiments, one or more of the pulmonary hypertension measurements described herein are measured after the patient has been treated with an ActRII polypeptide disclosed herein for 12 weeks. In some embodiments, one or more of the pulmonary hypertension measurements described herein are measured after the patient has been treated with an ActRII polypeptide disclosed herein for 16 weeks. In some embodiments, one or more of the pulmonary hypertension measurements described herein are measured after the patient has been treated with an ActRII polypeptide disclosed herein for 20 weeks. In some embodiments, one or more of the pulmonary hypertension measurements described herein are measured after the patient has been treated with an ActRII polypeptide disclosed herein for 22 weeks. In some embodiments, one or more of the pulmonary hypertension measurements described herein are measured after the patient has been treated with an ActRII polypeptide disclosed herein for 24 weeks. In some embodiments, one or more of the pulmonary hypertension measurements described herein are measured after the patient has been treated with an ActRII polypeptide disclosed herein for 26 weeks. In some embodiments, one or more of the pulmonary hypertension measurements described herein are measured after the patient has been treated with an ActRII polypeptide disclosed herein for 28 weeks. In some embodiments, one or more of the pulmonary hypertension measurements described herein are measured after the patient has been treated with an ActRII polypeptide disclosed herein for 48 weeks.

[0272] 5. Pharmaceutical Compositions and Modes of Administration In certain embodiments, the therapeutic method of the present disclosure comprises administering the composition systemically or locally as an implant or device.When administered, the therapeutic composition for use in the present disclosure is substantially pyrogen-free or in a physiologically acceptable form that is pyrogen-free.The therapeutically useful agent other than ActRII polypeptide, which may optionally be included in the above composition, can be administered simultaneously or sequentially with the subject compound in the method disclosed herein.

[0273] Typically, the protein therapeutic agent disclosed herein is administered parenterally, particularly intravenously or subcutaneously. Pharmaceutical compositions suitable for parenteral administration may include one or more ActRII polypeptides in combination with one or more pharma- ceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be reconstituted immediately before use into sterile injectable solutions or dispersions, and may include antioxidants, buffers, bacteriostatic agents, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents. Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present disclosure include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants. The formulations may be presented in unit-dose or multi-dose sealed containers, such as ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of a sterile liquid vehicle for injection, e.g., water, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets of the kind described herein.

[0274] The compositions and preparations may be provided in a pack or dispenser device that may contain one or more unit dosage forms containing the active ingredient, if desired.The pack may comprise, for example, metal or plastic foil, such as a blister pack.The pack or dispenser device may be accompanied by instructions for administration.

[0275] Furthermore, the composition can be encapsulated or injected into a form for delivery to target tissue site.In certain embodiments, the composition of the present invention can deliver one or more therapeutic compounds (e.g., ActRII polypeptide) to target tissue site, provide structure for developing tissue, and optimally include a matrix that can be resorbed in the body.For example, the matrix can provide sustained release of ActRII polypeptide.Such a matrix can be formed of materials currently used for other implantation medical applications.

[0276] The choice of matrix material is based on biocompatibility, biodegradability, mechanical properties, cosmetic appearance and interface properties. The particular application of the subject composition will define the appropriate formulation. Potential matrices for the composition can be biodegradable and chemically defined calcium sulfate, tricalcium phosphate, hydroxyapatite, polylactic acid and polyanhydrides. Other potential materials are biodegradable and biologically well defined, such as bone or dermal collagen. Further matrices are composed of pure proteins or extracellular matrix components. Other potential matrices are non-biodegradable and chemically defined, such as sintered hydroxyapatite, bioglass, aluminates or other ceramics. Matrices may be composed of combinations of any of the above types of materials, such as polylactic acid and hydroxyapatite or collagen and tricalcium phosphate. Bioceramics may vary in composition, such as calcium phosphate-aluminates, and processing may be modified to change pore size, particle size, particle shape, and biodegradability.

[0277] In certain embodiments, the methods of the invention can be administered orally, for example, in the form of capsules, cachets, pills, tablets, lozenges (using flavored bases, usually sucrose and acacia or tragacanth), powders, granules, each containing a predetermined amount of the agent as an active ingredient, or as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as a pastille (using an inert base such as gelatin and glycerin, or sucrose and acacia), and / or as a mouthwash, etc. The agent may also be administered as a bolus, electuary, or paste.

[0278] In solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules, etc.), one or more therapeutic compounds of the invention can be mixed with one or more pharma- ceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants, such as glycerol; (4) disintegrants, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as glycerol; (5) glyceryl stearate, (6) glyceryl monostearate, (7) wetting agents, such as cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof; and (10) coloring agents. In the case of capsules, tablets and pills, the pharmaceutical compositions may also contain buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar and high molecular weight polyethylene glycols.

[0279] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.In addition to active ingredients, liquid dosage forms can contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan, and mixtures thereof.In addition to inert diluents, oral compositions can also contain adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavorings, coloring agents, fragrances and preservatives.

[0280] Suspensions may contain, in addition to the active compound, suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, and mixtures thereof.

[0281] The composition of the present invention may also contain adjuvants such as preservatives, wetting agents, emulsifiers and dispersants.Prevention of microbial activity can be ensured by including various antibacterial and antifungal agents, such as paraben, chlorobutanol, phenol sorbic acid, etc.It may also be desirable to include isotonic agents, such as sugar, sodium chloride, etc. in the composition.Furthermore, delayed absorption of injectable pharmaceutical forms can be brought about by including agents that delay absorption, such as aluminum monostearate and gelatin.

[0282] It is understood that the dosing regimen is determined by the attending physician, taking into consideration various factors that modify the action of the subject compounds of the present disclosure (e.g., ActRII polypeptide). Various factors include, but are not limited to, the age, sex, and diet of the patient, the severity of the disease, the number of administrations, and other clinical factors. In some cases, the dosage may vary depending on the type of matrix used for reconstitution and the type of compound in the composition. In some embodiments, the patient's hematological parameters can be monitored by periodic evaluation to determine whether the patient has higher than normal red blood cell and / or hemoglobin levels (e.g., hemoglobin levels >16.0 g / dL or hemoglobin levels >18.0 g / dL). In some embodiments, patients with higher than normal red blood cell and / or hemoglobin levels can receive delayed or reduced doses until the levels return to normal or acceptable levels.

[0283] The likelihood that a patient will have a hemoglobin level of more than 18 g / dL or an increase in hemoglobin of more than 2 g / dL may increase during initial treatment with an ActRII polypeptide. In certain embodiments, a dosing regimen can be used to prevent, ameliorate, or reduce adverse changes in hemoglobin levels. In some embodiments, the ActRII polypeptide of the present disclosure is administered using a dosing regimen. In some embodiments, the method includes administering to the patient a therapeutically effective amount of an ActRII polypeptide disclosed herein, comprising a first dose of said polypeptide of 0.1 mg / kg to 1.0 mg / kg for a first period, followed by a second dose of said polypeptide of 0.1 mg / kg to 1.0 mg / kg administered for a second period. In some embodiments, the method comprises administering to the patient a therapeutically effective amount of a dosing regimen of an ActRII polypeptide disclosed herein, comprising a first dose of said polypeptide of 0.1 mg / kg to 1.0 mg / kg for a first period of time, a second dose of said polypeptide of 0.1 mg / kg to 1.0 mg / kg administered for a second period of time, and then a third dose of said polypeptide of 0.1 mg / kg to 1.0 mg / kg administered for a third period of time. In some embodiments, the first dose of ActRII polypeptide is administered to the patient in an amount of about 0.2 mg / kg to about 0.4 mg / kg. In some embodiments, the first dose of ActRII polypeptide is administered to the patient in a dose of 0.3 mg / kg. In some embodiments, the second dose of ActRII polypeptide is administered to the patient in an amount of about 0.5 mg / kg to about 0.8 mg / kg. In some embodiments, the second dose of ActRII polypeptide is administered to the patient in a dose of 0.7 mg / kg. In some embodiments, the third dose of the ActRII polypeptide is administered to the patient in an amount of about 0.2 mg / kg to about 0.4 mg / kg. In some embodiments, the third dose of the ActRII polypeptide is administered to the patient in a dose of 0.3 mg / kg.

[0284] In some embodiments, the dosing regimen comprises administering a first dose of an ActRII polypeptide to the patient in an amount of 0.3 mg / kg, followed by administering a second dose of an ActRII polypeptide to the patient in an amount of 0.7 mg / kg. In some embodiments, the dosing regimen comprises administering a first dose of an ActRII polypeptide to the patient in an amount of 0.3 mg / kg, administering a second dose of an ActRII polypeptide to the patient in an amount of 0.7 mg / kg, and administering a third dose of an ActRII polypeptide to the patient in an amount of 0.3 mg / kg. In some embodiments, the second dose is greater than the first dose. In some embodiments, the first dose is greater than the second dose. In some embodiments, the third dose is greater than the second dose. In some embodiments, the second dose is greater than the third dose. In some embodiments, the first period of time is at least 3 weeks. In some embodiments, the second period of time is at least 3 weeks. In some embodiments, the third period of time is at least 3 weeks. In some embodiments, the second period of time is at least 21 weeks. In some embodiments, the second period of time is at least 45 weeks. In some embodiments, the second period of time is greater than the first period of time. In some embodiments, the third period of time is greater than the first period of time. In some embodiments, the third period of time is greater than the second period of time.

[0285] In some embodiments, the change in dosage between the first dose and the second dose is determined by the attending physician in consideration of various factors (e.g., hemoglobin level). In some embodiments, the change in dosage between the second dose and the third dose is determined by the attending physician in consideration of various factors (e.g., hemoglobin level). In some embodiments, the various factors include, but are not limited to, changes in the patient's hematological parameters over a period of time. In some embodiments, the patient's hematological parameters are monitored to determine whether the red blood cell level and / or hemoglobin level is higher than normal (e.g., hemoglobin level >16.0 g / dL or hemoglobin level >18.0 g / dL). In some embodiments, the patient's hematological parameters are monitored to determine whether the increase in hemoglobin level over a period of time is higher than normal (e.g., an increase in hemoglobin level of more than 2 g / dL in less than 3 weeks). In some embodiments, if one or more of the patient's hematological parameters are abnormal before or during treatment, the patient's dose of an ActRII polypeptide disclosed herein is reduced (e.g., the dose is reduced from 0.7 mg / kg to 0.3 mg / kg). In some embodiments, if one or more of the patient's hematological parameters are abnormal before or during treatment, the patient's dose of an ActRII polypeptide disclosed herein is maintained (e.g., maintained at 0.3 mg / kg to 0.7 mg / kg).

[0286] In some embodiments, the dosing regimen prevents, ameliorates, or reduces adverse effects of the ActRII polypeptide. In some embodiments, administration of the ActRII polypeptide by the dosing regimen provided herein results in a reduction in adverse side effects. In some embodiments, administration of the ActRII polypeptide by the dosing regimen provided herein reduces the likelihood of hemoglobin levels exceeding 18 g / dL during the first period. In some embodiments, administration of the ActRII polypeptide by the dosing regimen provided herein reduces the likelihood of hemoglobin levels exceeding 18 g / dL during the first three weeks of treatment. In some embodiments, administration of the ActRII polypeptide by the dosing regimen provided herein reduces the likelihood of hemoglobin levels increasing by more than 2 g / dL during the first period. In some embodiments, administration of the ActRII polypeptide by the dosing regimen provided herein reduces the likelihood of hemoglobin levels increasing by more than 2 g / dL during the first three weeks of treatment.

[0287] In some embodiments, the ActRII polypeptide of the present disclosure is administered at a dose range of 0.1 mg / kg to 2.0 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 0.1 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 0.2 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 0.3 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 0.4 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 0.5 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 0.6 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 0.7 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 0.8 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 0.9 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 1.0 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 1.1 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 1.2 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 1.3 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 1.4 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 1.5 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 1.6 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 1.7 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 1.8 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 1.9 mg / kg. In some embodiments, the ActRII polypeptide of the present disclosure is administered at 2.0 mg / kg.

[0288] In certain embodiments, the ActRII polypeptide of the present disclosure is administered once a day. In certain embodiments, the ActRII polypeptide of the present disclosure is administered twice a day. In certain embodiments, the ActRII polypeptide of the present disclosure is administered once a week. In certain embodiments, the ActRII polypeptide of the present disclosure is administered twice a week. In certain embodiments, the ActRII polypeptide of the present disclosure is administered three times a week. In certain embodiments, the ActRII polypeptide of the present disclosure is administered every two weeks. In certain embodiments, the ActRII polypeptide of the present disclosure is administered every three weeks. In certain embodiments, the ActRII polypeptide of the present disclosure is administered every four weeks. In certain embodiments, the ActRII polypeptide of the present disclosure is administered monthly.

[0289] In certain embodiments, the present invention also provides gene therapy for in vivo production of ActRII polypeptide.Such therapy will achieve its therapeutic effect by introducing ActRII polypeptide polynucleotide sequence into cells or tissues with the above-mentioned disorders.Delivery of ActRII polypeptide polynucleotide sequence can be achieved using recombinant expression vector, such as chimeric virus or colloidal dispersion system.Targeted liposome can be used for therapeutic delivery of ActRII polypeptide polynucleotide sequence.

[0290] Various viral vectors that can be utilized for gene therapy as taught herein include RNA viruses such as adenovirus, herpes virus, vaccinia, or preferably retrovirus. Preferably, the retroviral vector is a derivative of a murine or avian retrovirus. Examples of retroviral vectors that can insert a single foreign gene include, but are not limited to, Moloney murine leukemia virus (MoMuLV), Harvey murine sarcoma virus (HaMuSV), mouse mammary tumor virus (MuMTV) and Rous sarcoma virus (RSV). Some additional retroviral vectors can incorporate multiple genes. All of these vectors can transfer or incorporate genes for selection markers so that transduced cells can be identified and generated. Retroviral vectors can be made target specific, for example, by attaching sugars, glycolipids, or proteins. Targeting can be achieved by using antibodies. Those skilled in the art will recognize that specific polynucleotide sequences can be inserted into the retroviral genome or attached to the viral envelope to allow target specific delivery of retroviral vectors that contain ActRII polypeptides. In one embodiment, the vector targets bone or cartilage.

[0291] Alternatively, tissue culture cells can be directly transfected with plasmids encoding the retroviral structural genes gag, pol and env by conventional calcium phosphate transfection. These cells are then transfected with a vector plasmid containing the gene of interest. The resulting cells release the retroviral vector into the culture medium.

[0292] Another targeted delivery system for ActRII polypeptide polynucleotides is a colloidal dispersion system. Colloidal dispersion systems include macromolecular complexes, nanocapsules, microspheres, beads, and lipid-based systems, including oil-in-water emulsions, micelles, mixed micelles, and liposomes. One colloidal system of the present invention is a liposome. Liposomes are artificial membrane vesicles that are useful as delivery vehicles in vitro and in vivo. RNA, DNA, and intact virions can be encapsulated in the aqueous interior and delivered to cells in a biologically active form (see, for example, Fraley, et al., Trends Biochem. Sci., 6:77, 1981). Methods for efficient gene transfer using liposome vehicles are known in the art, see, for example, Mannino, et al., Biotechniques, 6:682, 1988. The composition of liposomes is usually a combination of phospholipids, usually combined with steroids, especially cholesterol. Other phospholipids or other lipids may also be used. The physical characteristics of liposomes depend on pH, ionic strength, and the presence of divalent cations.

[0293] Examples of lipids useful for liposome manufacture include phosphatidyl compounds, such as phosphatidylglycerol, phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, sphingolipids, cerebrosides, and gangliosides.Exemplary phospholipids include egg phosphatidylcholine, dipalmitoyl phosphatidylcholine and distearoyl phosphatidylcholine.Targeting of liposome is also possible, for example, based on organ specificity, cell specificity and organelle specificity, and is known in the art.

[0294] The present disclosure provides formulations that can be modified to include acids and bases to adjust the pH; and buffers to maintain the pH within narrow ranges. 6. Kit The present disclosure provides a kit comprising a lyophilized polypeptide and an injection device. In certain embodiments, the lyophilized polypeptide comprises an ActRII polypeptide (e.g., a polypeptide that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acids 30-110 of SEQ ID NO:1), or a fragment, functional variant, or modified form thereof. In certain embodiments, the lyophilized polypeptide binds to one or more ligands selected from the group consisting of activin A, activin B, and GDF11. In certain embodiments, the lyophilized polypeptide further binds to one or more ligands selected from the group consisting of BMP10, GDF8, and BMP6. In certain embodiments, the lyophilized polypeptide binds to activin and / or GDF11.

[0295] In some embodiments, the lyophilized polypeptide begins at a residue corresponding to any one of amino acids 21-30 of SEQ ID NO:1 (e.g., begins at any one of amino acids 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) and ends at a position corresponding to any one of amino acids 110-135 of SEQ ID NO:1 (e.g., amino acids 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 18 1, 2, 123, 124, 125, ...

Claims

1. A composition for use in a method of treating pulmonary hypertension associated with a lung disease, a) an ActRIIA polypeptide comprising an amino acid sequence that is at least 95% identical to the amino acid sequence corresponding to residues 30 to 110 of SEQ ID NO: 1; b) the Fc domain of an IgG1 immunoglobulin; and c) a linker domain comprising a fusion protein, wherein the linker domain is located between the ActRIIA polypeptide and the Fc domain of the IgG1 immunoglobulin, and wherein the method is a composition that reduces right ventricular systolic pressure (RVSP) by at least 10%.

2. The composition according to claim 1, wherein the ActRIIA polypeptide comprises the amino acid sequence of SEQ ID NO:

2.

3. The composition according to claim 1, wherein the linker domain is selected from the group consisting of TGGG (SEQ ID NO: 20), TGGGG (SEQ ID NO: 18), SGGGG (SEQ ID NO: 19), GGGGS (SEQ ID NO: 22), GGG (SEQ ID NO: 16), GGGG (SEQ ID NO: 17), and SGGG (SEQ ID NO: 21).

4. The composition according to claim 1, wherein the linker domain is TGGG (SEQ ID NO: 20).

5. The composition according to claim 1, wherein the Fc domain comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

32.

6. The composition according to claim 1, wherein the Fc domain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:

32.

7. The composition according to claim 1, wherein the Fc domain comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO:

32.

8. The composition according to claim 1, wherein the fusion protein comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

23.

9. The composition according to claim 1, wherein the fusion protein comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:

23.

10. The composition according to claim 1, wherein the fusion protein comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO:

23.

11. The composition according to claim 1, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:

23.

12. The composition according to claim 1, wherein the fusion protein comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

41.

13. The composition according to claim 1, wherein the fusion protein comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:

41.

14. The composition according to claim 1, wherein the fusion protein comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO:

41.

15. The composition according to claim 1, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:

41.

16. The composition according to claim 1, wherein the fusion protein is part of a homodimeric protein complex.

17. The composition according to claim 1, wherein the fusion protein is glycosylated.

18. The composition according to claim 1, wherein the fusion protein binds to one or more ligands selected from the group consisting of activin A, activin B, GDF11, BMP10, GDF8, and BMP6.

19. A composition for use in a method of treating pulmonary hypertension associated with obstructive lung disease, comprising: a) an ActRIIA polypeptide comprising an amino acid sequence that is at least 95% identical to the amino acid sequence corresponding to residues 30 to 110 of SEQ ID NO: 1; b) the Fc domain of IgG1 immunoglobulin; and c) a fusion protein comprising a linker domain and wherein the linker domain is located between the ActRIIA polypeptide and the Fc domain of the IgG1 immunoglobulin, said composition.

20. The composition according to claim 19, wherein the ActRIIA polypeptide comprises the amino acid sequence of SEQ ID NO:

2.

21. The composition according to claim 19, wherein the linker domain is selected from the group consisting of TGGG (SEQ ID NO: 20), TGGGG (SEQ ID NO: 18), SGGGG (SEQ ID NO: 19), GGGGS (SEQ ID NO: 22), GGG (SEQ ID NO: 16), GGGG (SEQ ID NO: 17), and SGGG (SEQ ID NO: 21).

22. The composition according to claim 19, wherein the linker domain is TGGG (SEQ ID NO: 20).

23. The composition according to claim 19, wherein the Fc domain comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

32.

24. The composition according to claim 19, wherein the Fc domain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:

32.

25. The composition according to claim 19, wherein the Fc domain comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO:

32.

26. The composition according to claim 19, wherein the fusion protein comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

23.

27. The composition according to claim 19, wherein the fusion protein comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:

23.

28. The composition according to claim 19, wherein the fusion protein comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO:

23.

29. The composition according to claim 19, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:

23.

29. The composition according to claim 19, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:

23. **Claim 30**: The composition according to claim 19, wherein the fusion protein comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

41. **Claim 31**: The composition according to claim 19, wherein the fusion protein comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:

41. **Claim 32**: The composition according to claim 19, wherein the fusion protein comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO:

41. **Claim 33**: The composition according to claim 19, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:

41. **Claim 34**: The composition according to claim 19, wherein the fusion protein is part of a homodimeric protein complex. **Claim 35**: The composition according to claim 19, wherein the fusion protein is glycosylated. **Claim 36**: The composition according to claim 19, wherein the obstructive lung disease is selected from the group consisting of chronic obstructive pulmonary disease (COPD), cystic fibrosis, asthma, emphysema, lymphangioleiomyomatosis, and chronic bronchitis. **Claim 37**: The composition according to claim 1 or 19, wherein the method prevents or reduces the progression of the pulmonary hypertension functional classification as recognized by the World Health Organization (WHO).

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