Treating cardiotoxicity and / or hypertension

Abstract

Methods and materials for treating or preventing cardiotoxicity and / or hypertension induced by one or more anti-cancer agents (e.g., tyrosine kinase inhibitor (TKI) -induced cardiotoxicity (e.g., sunitinib-induced cardiotoxicity) and / or TKI-induced hypertension (e.g., sunitinib-induced hypertension)) are provided herein. For example, provided herein are methods for using polypeptides (e.g., chimeric polypeptides) that include (a) one or more natriuretic peptides and (b) one or more angiotensin 1-7 (ANG1-7) polypeptides to treat or prevent cardiotoxicity and / or hypertension induced by one or more anti-cancer agents.

Description

[0001] Attorney Docket No. 07039-2323WO1

[0002] 2024-157

[0003] TREATING CARDIOTOXICITY AND / OR HYPERTENSION

[0004] CROSS-REFERENCE TO RELATED APPLICATIONS

[0005] This application claims the benefit of U.S. Patent Application Serial No. 63 / 704,286, filed on October 7, 2024. The disclosure of the prior application is considered part of, and is incorporated by reference in, the disclosure of this application.

[0006] SEQUENCE LISTING

[0007] This application contains a Sequence Listing that has been submitted electronically as an XML file named “07039-2323W01.xml.” The XML file, created on September 22, 2025, is 19,505 bytes in size. The material in the XML file is hereby incorporated by reference in its entirety.

[0008] TECHNICAL FIELD

[0009] This document relates to methods and materials for treating or preventing cardiotoxicity and / or hypertension induced by one or more anti-cancer agents (e.g., tyrosine kinase inhibitor (TKI) -induced cardiotoxicity (e.g., sunitinib-induced cardiotoxicity) and / or TKI-induced hypertension (e.g., sunitinib-induced hypertension)). For example, this document provides methods for using polypeptides (e.g., chimeric polypeptides) that include (a) one or more natriuretic peptides and (b) one or more angiotensin 1-7 (ANG1-7) polypeptides to treat or prevent cardiotoxicity and / or hypertension induced by one or more anti-cancer agents.

[0010] BACKGROUND

[0011] Tyrosine kinase inhibitors (TKIs) exhibit potent anti-tumor effects but are associated with detrimental cardiovascular side effects (Gschwind et al., Nat. Rev. Cancer, 4:361-370 (2004); Shyam Sunder et al., Sigi. Transd. Targ. Then, 8:262 (2023); Chen et al., Circulation. 118:84-95 (2008); and Force et al., Nat. Rev. Cancer, 7:332-344 (2007)). Sunitinib, a multitargeted TKI. boosts survival in metastatic renal-cell carcinoma and gastrointestinal stromal tumors (Faivre et al., J. Clin. Oncol., 24:25-35 (2006); and Demetri et al., Lancet, 368: 1329-1338 (2006)). However, its use often triggers hypertension and cardiotoxicity, leading to left ventricular (LV) dysfunction and heart failure (HF) (Chen et al., Circulation, 118:84-95 (2008); Cohen et al.. Hypertension Attorney Docket No. 07039-2323WO1

[0012] 2024-157

[0013] 80:e46-e57 (2023); Chu et al., Lancet, 370:2011-2019 (2007); Ewer et al., Nat. Clin. Pract. Cardiovasc. Med., 5:364-365 (2008); and Mirabito Colafella et al.. Cardiovasc. Res., 116: 1779-1790 (2020)). There are no FDA-approved therapies available to target these adverse cardiovascular side effects induced by TKI such as sunitinib.

[0014] SUMMARY

[0015] This document provides methods and materials for treating or preventing cardiotoxicity and / or hypertension induced by one or more anti-cancer agents (e.g.. TKI- induced cardiotoxicity (e.g., sunitinib-induced cardiotoxicity) and / or TKI-induced hypertension (e.g., sunitinib-induced hypertension)). For example, this document provides methods for using polypeptides (e.g., chimeric polypeptides) that include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides (also be referred to herein as NPA7 polypeptides) to treat or prevent cardiotoxicity and / or hypertension induced by one or more anti-cancer agents. In some cases, one or more NPA7 polypeptides can be administered to a mammal (e.g., a human) having, or at risk of developing, cardiotoxicity and / or hypertension induced by one or more anti-cancer agents to treat or prevent the cardiotoxicity and / or hypertension. As demonstrated herein, NPA7 polypeptides can simultaneously activate a natriuretic peptide / particulate guany lyl cyclase-A receptor / cGMP (NP / pGC-A / cGMP) pathway and an angiotensini.y / Mas receptor / cAMP (ANGi-7 / MasR / cAMP) pathway, thereby promoting the release of nitric oxide (NO) in endothelial cells and protecting those cells from oxidative stress, mitochondrial fragmentation, hypertrophy, and cell death in human cardiomyocytes (HCMs).

[0016] Having the ability to treat or prevent cardiotoxicity and / or hypertension induced by one or more anti -cancer agents (e.g., TKI-induced cardiotoxicity (e.g., sunitinib- induced cardiotoxicity) and / or TKI-induced hypertension (e.g., sunitinib-induced hypertension)) as described herein (e.g.. by administering one or more NPA7 polypeptides) provides a unique and unrealized opportunity minimize or even prevent adverse effects of commonly used anti-cancer agents (e.g., one or more TKIs such as sunitinib), thus allowing patients a significant clinic benefit of being treated with lower risk of adverse cardiovascular side effects induced by the anti-cancer agents. Attorney Docket No. 07039-2323WO1

[0017] 2024-157

[0018] In general, one aspect of this document features methods for delaying or preventing cardiotoxicity in a mammal having been administered an anti -cancer agent. The methods can include, or consist essentially of, administering a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1 -7 peptide, a nucleic acid encoding said polypeptide, or a composition comprising said polypeptide, said nucleic acid, or both to said mammal. The mammal can be a human. The anti-cancer agent can be aTK The anti-cancer agent can be sunitinib. sorafenib, dasatinib, lenvatinib, imatinib, nilotinib, bosutinib, or ponatinib. The natriuretic peptide can be an atrial natriuretic peptide (ANP), a brain natriuretic peptide (BNP), a C-type natriuretic peptide (CNP), a urodilatin (URO) peptide, or a Dendroaspis natriuretic peptide (DNP). The natriuretic peptide can comprise, consist essentially of, or consist of any one of SEQ ID NOs:2-6. The ANG1-7 peptide can comprise, consist essentially of, or consist of the amino acid sequence set forth in SEQ ID NO:7. The polypeptide can comprise, consist essentially of, or consist of the amino acid sequence set forth in SEQ ID NO: 1. The administering can be subcutaneous, intravenous, or oral administration.

[0019] In another aspect, this document features methods for delaying or preventing cardiotoxicity in a mammal scheduled to receive an anti-cancer agent. The methods can include, or consist essentially of, administering a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide, a nucleic acid encoding said polypeptide, or a composition comprising said polypeptide, said nucleic acid, or both to said mammal. The mammal can be a human. The anti-cancer agent can be a TKI. The anti-cancer agent can be sunitinib, sorafenib, dasatinib, lenvatinib, imatinib, nilotinib, bosutinib, or ponatinib. The natriuretic peptide can be an atrial natriuretic peptide (ANP), a brain natriuretic peptide (BNP), a C-type natriuretic peptide (CNP). a urodilatin (URO) peptide, or a Dendroaspis natriuretic peptide (DNP). The natriuretic peptide can comprise, consist essentially of, or consist of any one of SEQ ID NOs:2-6. The ANG1-7 peptide can comprise, consist essentially of, or consist of the amino acid sequence set forth in SEQ ID NO:7. The polypeptide can comprise, consist essentially of, or consist of the amino acid sequence set forth in SEQ ID NO: 1. The administering can be subcutaneous, intravenous, or oral administration.

[0020] In another aspect, this document features methods for delaying or preventing hypertension in a mammal having been administered an anti-cancer agent. The methods Attorney Docket No. 07039-2323WO1

[0021] 2024-157 can include, or consist essentially of, administering a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide, a nucleic acid encoding said polypeptide, or a composition comprising said polypeptide, said nucleic acid, or both to said mammal. The mammal can be a human. The anti-cancer agent can be a TKI. The anti-cancer agent can be sunitinib, sorafenib, dasatinib, lenvatinib, imatinib, nilotinib, bosutinib, or ponatinib. The natriuretic peptide can be an ANP, a BNP, a CNP, a URO peptide, or a DNP. The natriuretic peptide can comprise, consist essentially of, or consist of any one of SEQ ID NOs:2-6. The ANG1-7 peptide can comprise, consist essentially of, or consist of the amino acid sequence set forth in SEQ ID NO: 7. The polypeptide can comprise, consist essentially of, or consist of the amino acid sequence set forth in SEQ ID NO: 1. The administering can be subcutaneous, intravenous, or oral administration.

[0022] In another aspect, this document features methods for delaying or preventing hypertension in a mammal scheduled to receive an anti-cancer agent. The methods can include, or consist essentially of, administering a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide, a nucleic acid encoding said polypeptide, or a composition comprising said polypeptide, said nucleic acid, or both to said mammal. The mammal can be a human. The anti-cancer agent can be a TKI. The anti-cancer agent can be sunitinib, sorafenib, dasatinib, lenvatinib, imatinib, nilotinib, bosutinib, or ponatinib. The natriuretic peptide can be an ANP, a BNP, a CNP, a URO peptide, or a DNP. The natriuretic peptide can comprise, consist essentially of, or consist of any one of SEQ ID NOs:2-6. The ANG1-7 peptide can comprise, consist essentially of, or consist of the amino acid sequence set forth in SEQ ID NO: 7. The polypeptide can comprise, consist essentially of, or consist of the amino acid sequence set forth in SEQ ID NO: 1. The administering can be subcutaneous, intravenous, or oral administration.

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used to practice the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the Attorney Docket No. 07039-2323WO1

[0024] 2024-157 present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0025] The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention w ill be apparent from the description and drawings, and from the claims.

[0026] BRIEF DESCRIPTION OF THE DRAWINGS

[0027] FIG. 1. NPA7 polypeptides can prevent sunitinib-induced HTN as well as protect against cardiotoxicity' through suppressing mitochondrial fragmentation via the NF-E2- related factor 2 (Nrf2) pathway. The mechanism of sunitinib-induced HTN is complex and involves a reduction of nitric oxide (NO) and cGMP levels as w ell as endothelin (ET)-l up-regulation, while cardiotoxicity has been associated with mitochondrial dysfunction and oxidative stress.

[0028] FIG. 2. An amino acid sequence (SEQ ID NO:1) of an exemplary' NPA7 polypeptide having a 22 -amino acid (AA) sequence of a BNP peptide fused to a 7-AA sequence of an ANG1-7 peptide.

[0029] FIGS. 3A-3B. Generation of cGMP (FIG 3A) or cAMP (FIG. 3B) in HCMs stimulated by aNPA7 polypeptide. ***P<0.001, **P<0.05.

[0030] FIG. 4. NPA7 polypeptides significantly reduced systolic blood pressure in spontaneously hypertensive rats.

[0031] FIG. 5. NPA7 polypeptides promoted the release of NO in endothelial cells. NO generation was measured using a chemiluminescence assay. ***P<0.001.

[0032] FIGS. 6A-6D. NPA7 polypeptides acted as aNRF2 activator and safeguarded against oxidative stress via the KEAP1-NRF2 pathw ay in HCMs. FIG. 6A includes representative immunofluorescent images for dihydroethidium (DHE) staining. FIG. 6B is a graph plotting quantification of DHE staining intensity. Scale bar=100 pm. FIG. 6C is a graph plotting relative levels of mRNAs of NRF2 target genes, as determined by qRT- PCR analysis. FIG. 6D shows a representative Western blot showing levels of KEAP1 polypeptides and NRF2 polypeptides. Attorney Docket No. 07039-2323WO1

[0033] 2024-157

[0034] FIGS. 7A-7B. NPA7 polypeptides inhibited sunitinib-induces reactive oxygen species (ROS) production in HCMs. Immunofluorescent images (FIG. 7A) and intensity quantification (FIG. 7B) for DHE staining are shown. Scale bar=50 pm. ***P<0.001.

[0035] FIGS. 8A-8B. Sunitinib induced oxidative stress mediated mitochondrial fragmentation. HCMs pretreated with the ROS inhibitor N-acetyl-L-cysteine (NAC; 5 mM) were exposed to sunitinib (10 pM) for 24 hours, and the immunofluorescence of mitochondria membrane protein TOM20 was detected (FIG. 8A) and mitochondrial length was analyzed (FIG. 8B). Scale bar=10 pm.

[0036] FIGS. 9A-9C. Sunitinib induced hypertrophy in HCMs. Phalloidin staining (FIG. 9A), cell surface area quantification (FIG. 9B), and qRT-PCR analysis of NPPA and MYH7 (FIG. 9C) of HCMs in the presence of vehicle, 10 pM sunitinib, or sunitinib + NAC for 24 hours are shown. Scale bar=50 pm. ***P<0.001.

[0037] FIGS. 10A-10B. Sunitinib increased blood pressure in Wistar Kyoto (WKY) rats. Systolic blood pressure (FIG. 10A), NPPA (gene encoding ANP) mRNA levels, and NPPB (gene encoding BNP) mRNA levels (FIG. 10B) were measured in the left ventricle after oral gavage of sunitinib. ***P<0.001, **P<0.05.

[0038] FIGS. 11A-11C. NPA7 polypeptides suppressed sunitinib-induced hypertrophy in HCMs. Phalloidin staining (FIG. 11 A), cell surface area quantification (FIG. 11B), and qRT-PCR analysis of NPPA (FIG. 11C) of HCMs in the presence of vehicle, 10 pM sunitinib+PBS, or sunitinib + NPA7 for 24 hours are shown. Scale bar=50 pm. ***P<0.001.

[0039] FIG. 12. NPA7 polypeptides inhibited sunitinib-induced mitochondrial fragmentation of HCMs. The immunofluorescence of MitoTracker™ dye staining was performed.

[0040] FIG. 13. NPA7 polypeptides inhibited sunitinib-induced reduction of mitofusin-1 fission (MFN1) polypeptides and inhibited sunitinib-induced increase of mitochondrial fission factor (MFF) polypeptide levels in HCMs treated with sunitinib.

[0041] FIG. 14. NPA7 polypeptides promoted KEAP1 degradation and NRF2 activation in HCMs treated with sunitinib.

[0042] FIG. 15. NPA7 inhibited sunitinib-induced lactate dehydrogenase (LDH) cytotoxicity in HCMs. LDH cytotoxicity was quantified. *** PO.OOl. Attorney Docket No. 07039-2323WO1

[0043] 2024-157

[0044] FIG. 16. NPA7 rescued the reduction in ATP levels induced by sunitinib in HCMs, in a dose-dependent manner. *** PO.OOl. ** P<0.05.

[0045] FIGS. 17A-17B. NPA7 rescued the reduction in mitochondrial membrane potential (MMP) induced by sunitinib in HCMs, in a dose-dependent manner. Immunofluorescent images (FIG. 17A) and intensity quantification (FIG. 17B) for TMRM staining. Scale bar=100 [im. *** PO.OOl. ** P .05.

[0046] FIGS. 18A-18B. NPA7 inhibited sunitinib-induced production of mitochondrial ROS in HCMs. Immunofluorescent images (FIG. 18 A) and intensity quantification (FIG. 18B) for MitoSox staining. Scale bar=100 pm. ***P<0.001.

[0047] DETAILED DESCRIPTION

[0048] This document provides methods and materials for treating or preventing cardiotoxicity and / or hypertension induced by one or more anti-cancer agents (e.g.. TKI- induced cardiotoxicity (e.g., sunitinib-induced cardiotoxicity) and / or TKI-induced hypertension (e.g., sunitinib-induced hypertension)). In some cases, this document provides methods and materials for using one or more NPA7 polypeptides (e.g., polypeptides (e.g., chimeric polypeptides) that include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) to treat or prevent cardiotoxicity in a mammal (e.g., a human) currently being administered or scheduled to be administered one or more anti-cancer agents (e.g., one or more TKIs such as sunitinib). For example, one or more NPA7 polypeptides can be administered to a mammal (e.g., a human) having, or at risk of developing, cardiotoxicity induced by one or more anti-cancer agents to treat the mammal. In some cases, this document provides methods and materials for using one or more NPA7 polypeptides to treat or prevent hypertension in a mammal (e.g., a human) currently being administered or scheduled to be administered one or more anti-cancer agents (e.g., one or more TKIs such as sunitinib). For example, one or more NPA7 polypeptides can be administered to a mammal (e.g., a human) having, or at risk of developing, TKI-induced hypertension (e.g., sunitinib-induced hypertension) to treat the mammal.

[0049] Cardiotoxicity and / or hypertension can be induced by any type of anti -cancer agent. In some cases, an anti-cancer agent that can induce cardiotoxicity and / or hypertension can be a kinase inhibitor such as aTKI, a Bruton tyrosine kinase inhibitor. Attorney Docket No. 07039-2323WO1

[0050] 2024-157 or a RET kinase inhibitor. Examples of anti-cancer agents that can induce cardiotoxicity and / or hypertension include, without limitation, sunitinib, sorafenib, dasatinib, lenvatinib, imatinib, nilotinib, bosutinib, and ponatinib.

[0051] Any type of mammal can be treated as described herein (e.g., to treat or prevent cardiotoxicity and / or hypertension induced by one or more anti-cancer agents). Examples of mammals that can be treated with one or more NPA7 polypeptides provided herein (e.g., one or more polypeptides (e.g., one or more chimeric polypeptides) that each include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) as described herein include, without limitation, humans, non-human primates (e.g., monkeys), dogs, cats, horses, cows, pigs, sheep, rabbits, mice, rats, and goats. For example, a human having, or at risk of developing, cardiotoxicity and / or hypertension induced by one or more anti-cancer agents can be treated with one or more NPA7 polypeptides as described herein. In some cases, a mammal to be treated as described herein can have cancer. For example, a mammal to be treated as described herein can have cancer and can have previously been administered, be currently taking, and / or schedule to take one or more anti-cancer agents (e.g., one or more TKIs such as sunitinib) to treat the cancer.

[0052] In some cases, the methods provided herein can include identifying a mammal (e.g., a human) as developing cardiotoxicity7and / or hypertension induced by one or more anti-cancer agents (e.g., TKI -induced cardiotoxicity (e.g., sunitinib-induced cardiotoxicity) and / or TKI-induced hypertension (e.g., sunitinib-induced hypertension)). For example, the methods provided herein can include identifying a mammal being administered one or more anti-cancer agents as having one or more early signs of cardiotoxicity. Any appropriate method can be used to identify a mammal as developing cardiotoxicity (e.g., as having one or more early signs of cardiotoxicity) induced by one or more anti-cancer agents. In some cases, detection of cardiac biomarkers (e.g., Troponin I or T (Tnl or TnT), BNP / N-terminal pro-B-type natriuretic peptide (NT-proBNP), and myocardial enzymes such as LDH and CK), echocardiography (e.g., to monitor ventricular function such as overall left ventricular function as measured by left ventricular ejection fraction (LVEF). fractional shortening, wall stress, and global longitudinal strain (GLS)), electrocardiograms (ECGs), cardiac magnetic resonance (CMR) (e.g., to determine left ventricular ejection fraction (LVEF), volumes, strain, Attorney Docket No. 07039-2323WO1

[0053] 2024-157 injury', and fibrosis), PET / CT imaging, and / or MUGA scans can be used to identify a human or other mammal as developing cardiotoxicity induced by one or more anti -cancer agents. For example, the methods provided herein can include identifying a mammal being administered one or more anti-cancer agents as having one or more early signs of hypertension. Any appropriate method can be used to identify' a mammal as developing hypertension (e.g., as having one or more early signs of hypertension) induced by one or more anti-cancer agents. In some cases, blood pressure readings, electrocardiograms (ECGs), and / or kidney function tests (e.g., blood tests for blood urea nitrogen (BUN) levels, serum creatinine levels, and estimated glomerular filtration rate (eGFR), and urinalysis for proteinuria) can be used to identify' a human or other mammal as developing hypertension induced by one or more anti-cancer agents.

[0054] In some cases, the methods provided herein can include adjusting the dosing of one or more anti-cancer agents based on whether a mammal (e g., a human) is developing cardiotoxicity and / or hypertension induced by the one or more anti-cancer agents (e.g., TKI-induced cardiotoxicity (e.g., sunitinib-induced cardiotoxicity) and / or TKI-induced hypertension (e.g., sunitinib-induced hypertension)). In some cases, the dosing of one or more anti-cancer agents can be adjusted based, at least in part, on whether a mammal (e.g., a human) is identified as developing cardiotoxicity induced by the one or more anticancer agents (e.g., TKI-induced cardiotoxicity (e.g., sunitinib-induced cardiotoxicity). For example, when a mammal being administered one or more anti-cancer agents is identified as having one or more early signs of cardiotoxicity, the dosing of the one or more anti-cancer agents can be reduced. In another example, when a mammal being administered one or more anti-cancer agents is identified as not having one or more early signs of cardiotoxicity, the dosing of the one or more anti-cancer agents can be maintained or increased. In some cases, the dosing of one or more anti-cancer agents can be adjusted based, at least in part, on whether a mammal (e.g., a human) is identified as developing hypertension induced by the one or more anti-cancer agents (e.g., TKI- induced hypertension (e.g., sunitinib-induced hypertension). For example, when a mammal being administered one or more anti-cancer agents is identified as having one or more early signs of hypertension, the dosing of the one or more anti-cancer agents can be reduced. In another example, when a mammal being administered one or more anti- Attorney Docket No. 07039-2323WO1

[0055] 2024-157 cancer agents is identified as not having one or more early signs of hypertension, the dosing of the one or more anti-cancer agents can be maintained or increased.

[0056] A mammal having, or at risk of developing, cardiotoxicity and / or hypertension induced by one or more anti-cancer agents (e.g., TKI-induced cardiotoxicity (e.g., sunitinib-induced cardiotoxicity) and / or TKI-induced hypertension (e.g., sunitinib- induced hypertension)) can be administered or instructed to self-administer one or more (e.g., one, two, three, four, or more) NPA7 polypeptides.

[0057] In some cases, a mammal (e.g., a human) being administered one or more anticancer agents (e.g., one or more TKIs such as sunitinib) can be concurrently administered (or instructed to self-administer) one or more (e.g., one, two, three, four, or more) NPA7 polypeptides. For example, one or more anti-cancer agents and one or more NPA7 polypeptides can be administered to a mammal to treat or prevent cardiotoxicity and / or hypertension induced by one or more anti-cancer agents. In some cases, one or more anticancer agents and one or more NPA7 polypeptides can be administered in a single composition including both the one or more anti-cancer agents and the one or more NPA7 polypeptides. In some cases, one or more anti-cancer agents and one or more NPA7 polypeptides can be administered as separate compositions that are concurrently administered.

[0058] In some cases, a mammal (e.g., a human) scheduled to be administered one or more anti-cancer agents (e.g., one or more TKIs such as sunitinib) can be administered (or instructed to self-administer) one or more (e.g., one. two, three, four, or more) NPA7 polypeptides. For example, one or more NPA7 polypeptides can be administered to a mammal scheduled to be administered one or more anti-cancer agents to prevent the one or more anti -cancer agents from inducing cardiotoxicity and / or hypertension.

[0059] A NPA7 polypeptide provided herein (e.g.. a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) can include any appropriate one or more natriuretic peptides. A natriuretic peptide that can be included in a polypeptide provided herein can be any peptide that can activate the GC-A / cGMP pathway. For example, a natriuretic peptide can be a GC-A activator. Examples of natriuretic peptides include, without limitation, atrial natriuretic peptides (ANPs), brain natriuretic peptides (BNPs), C-type natriuretic peptides (CNPs), urodilatin (URO) peptides, and Dendroaspis natriuretic peptides (DNPs). Attorney Docket No. 07039-2323WO1 2024-157

[0060] A natriuretic peptide that can be included in a NPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) can be any appropriate length (e.g., can include any number of amino acids). In some cases, a natriuretic peptide can be from about 15 amino acids in length to about 60 amino acids in length (e.g., from about 15 to about 50, from about 15 to about 40, from about 15 to about 30, from about 25 to about 60, from about 35 to about 60, from about 45 to about 60, from about 20 to about 50, from about 30 to about 40. from about 20 to about 30, from about 25 to about 35, from about 35 to about 45, from about 40 to about 50, or from about 45 to about 55 amino acids in length).

[0061] A natriuretic peptide that can be included in a NPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) can have any appropriate amino acid sequence. Natriuretic peptides ty pically have a ring structure resulting from a disulfide bond between two cysteine residues present within the peptide. In some cases, a natriuretic peptide can comprise, consist essentially of, or consist of an amino acid sequence set forth in TABLE 1.

[0062] A natriuretic peptide that consists essentially of an amino acid sequence set forth in TABLE 1 can include the amino acid sequence set forth in any one of SEQ ID NOs:2-6 with zero, one, or two amino acid substitutions within the articulated sequence of the sequence identifier (e.g., any one of SEQ ID NOs:2-6), with zero, one, two, three, four, or five amino acid residues preceding the articulated sequence of the sequence identifier (e.g., any one of SEQ ID NOs:2-6), and / or with zero, one. two, three, four, or five amino acid residues following the articulated sequence of the sequence identifier (e.g., any one of SEQ ID NOs:6), provided that the natriuretic peptide retains at least some natriuretic peptide activity (e.g.. the ability to activate the GC-A / cGMP pathway).

[0063] TABLE 1. Exemplary natriuretic peptide amino acid sequences with cysteine residues involved in the disulfide bond underlined. Attorney Docket No. 07039-2323WO1

[0064] 2024-157

[0065] In some cases, a natriuretic peptide that can be included in a NPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) can have an amino acid set forth in the National Center for Biotechnology Information (NCBI) databases at, for example, accession no. NM_002521 (e.g., version NM_002521.3), accession no. NM_008726 (e.g., version NM_008726.6), accession no. NM_006172 (e.g., version NM_006172.4).

[0066] In some cases, a natriuretic peptide that can be included in a NPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) can be as described elsewhere (see, e.g., Meems et al., Hypertension 73:900-909 (2019); and International Patent Application Publication No. WO 2013 / 032784).

[0067] A NPA7 polypeptide provided herein (e.g.. a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) can include any appropriate ANG1 -7 peptide. An ANG1-7 peptide that can be included in aNPA7 polypeptide provided herein can be any polypeptide that can activate the ANGi- 7 / MasR / cAMP pathway. For example, an ANG1-7 peptide can be a MAS activator.

[0068] An ANG1-7 peptide that can be included in a NPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) can have any appropriate amino acid sequence. In some cases, an ANG1-7 peptide can comprise, consist essentially of, or consist of the amino acid sequence set forth in SEQ ID NO: 7 below.

[0069] DRVYIHP (SEQ ID NO: 7)

[0070] In some cases, an ANG1-7 peptide that can be included in a NPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) can be as described elsewhere (see, e.g., Medina et al., Am. J. Hyperlens.. 32(12): 1133-1142 (2019); and Santos, Hypertension. 63: 1138— 1147 (2014)).

[0071] In some cases, a NPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) can have an amino acid sequence set forth in SEQ ID NO: 1 (see, e.g.. FIG. 2). In some cases, a NPA7 polypeptide can be a polypeptide that includes (a) one or more natriuretic Attorney Docket No. 07039-2323WO1

[0072] 2024-157 peptides and (b) one or more ANG1-7 peptides as described in International Patent Application Publication No. WO 2013 / 032784. See, e.g., FIGS. 1-20 and 29-32.

[0073] Any appropriate method can be used to obtain aNPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides). For example, aNPA7 polypeptide provided herein can be obtained by synthesizing the polypeptide of interest using appropriate polypeptide synthesizing techniques.

[0074] This document also provides nucleic acid molecules that can encode a NPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides). For example, this document provides nucleic acid molecules encoding a NPA7 polypeptide having the amino acid sequence set forth in SEQ ID NOT (see, e.g., FIG 2).

[0075] In some cases, a nucleic acid that can encode a NPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) can be in the form of a vector (e.g., a viral vector or a non-viral vector).

[0076] When a vector including nucleic acid that can encode a NPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) is a viral vector, any appropriate viral vector can be used. Examples viral vectors include, without limitation, Ad vectors, AAV vectors. HSV vectors, retrovirus vectors, and lentivirus vectors.

[0077] When a vector that can encode a NPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1- 7 peptides) is a non-viral vector, any appropriate non-viral vector can be used. In some cases, a non-viral vector can be an expression plasmid (e.g.. a cDNA expression vector).

[0078] In addition to nucleic acid that can encode a NPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides), a vector (e.g., a viral vector or a non-viral vector) can contain one or more regulatory elements operably linked to the nucleic acid that can encode a NPA7 polypeptide provided herein. Such regulatory elements can include promoter sequences, enhancer sequences, response elements, signal peptides, internal ribosome entry sequences, poly adenylation signals, terminators, and inducible elements that modulate Attorney Docket No. 07039-2323WO1

[0079] 2024-157 expression (e.g., transcription or translation) of a nucleic acid. The choice of regulatory' element(s) that can be included in a vector depends on several factors, including, without limitation, inducibility. targeting, and the level of expression desired. For example, a promoter can be included in a vector to facilitate transcription of a nucleic acid encoding a nucleoporin polypeptide. A promoter can be a naturally occurring promoter or a recombinant promoter. A promoter can be ubiquitous or inducible (e.g., in the presence of tetracycline), and can affect the expression of a nucleic acid encoding a polypeptide in a general or tissue-specific manner. Examples of promoters that can be used to drive expression of a NPA7 polypeptide provided herein in cells include, without limitation, CMV promoters, EF-la promoters, and SV40 promoters. As used herein, “operably linked’" refers to positioning of a regulatory element relative to a nucleic acid encoding a polypeptide in such a way as to permit or facilitate expression of the encoded polypeptide. For example, a vector can contain a promoter and nucleic acid that can encode a NPA7 polypeptide provided herein. In this case, the promoter is operably linked to a nucleic acid that can encode a NPA7 polypeptide provided herein such that it drives expression of the NPA7 polypeptide in cells.

[0080] Nucleic acid that can encode a NPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1- 7 peptides) can be produced by techniques including, without limitation, common molecular cloning, polymerase chain reaction (PCR), chemical nucleic acid synthesis techniques, and combinations of such techniques. For example, PCR or RT-PCR can be used with oligonucleotide primers designed to amplify nucleic acid (e.g., genomic DNA or RNA) that can encode aNPA7 polypeptide provided herein (e.g., a polypeptide that includes (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides).

[0081] One or more NPA7 polypeptides provided herein (e.g., one or more polypeptides (e.g., one or more chimeric polypeptides) that each include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein can be formulated into a composition (e.g., a pharmaceutically acceptable composition) for administration to a mammal having, or at risk of developing, cardiotoxicity and / or hypertension induced by one or more anticancer agents (e g., TKI-induced cardiotoxicity (e.g., sunitinib-induced cardiotoxicity) and / or TKI-induced hypertension (e.g., sunitinib-induced hypertension)). For example, a Attorney Docket No. 07039-2323WO1

[0082] 2024-157 therapeutically effective amount of one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) can be formulated together with one or more pharmaceutically acceptable carriers (additives) and / or diluents. A pharmaceutical composition can be formulated for administration in solid or liquid form including, without limitation, sterile solutions, suspensions, sustained-release formulations, tablets, capsules, pills, powders, and granules.

[0083] A composition (e.g., a pharmaceutically acceptable composition) including one or more NPA7 polypeptides provided herein (e.g., one or more polypeptides (e.g., one or more chimeric polypeptides) that each include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein can be administered locally or systemically. A composition containing one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) can be designed for oral, parenteral (including subcutaneous, intramuscular, intravenous, and intradermal), or inhaled administration. For example, a composition containing one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) can be administered systemically by an oral administration to or inhalation by a mammal (e.g., a human). When being administered orally, a composition containing one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) can be in the form of a pill, tablet, or capsule.

[0084] In some cases, one or more (e.g., one, two, three, four, or more) NPA7 polypeptides provided herein (e.g.. one or more polypeptides (e g., one or more chimeric polypeptides) that each include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein can be used to delay the progression of or prevent the onset of one or more symptoms of cardiotoxicity induced by one or more anti-cancer agents (e.g., TKI-induced cardiotoxicity (e.g., sunitinib-induced cardiotoxicity). For example, one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) can be administered to a mammal (e.g., a human) in need thereof (e.g., a human being administered of scheduled to be Attorney Docket No. 07039-2323WO1

[0085] 2024-157 administered one or more anti-cancer agents) to delay the progression of or prevent the onset of one or more symptoms of the cardiotoxicity. Examples of symptoms of cardiotoxicity (e.g., cardiotoxicity induced by one or more anti-cancer agents) include, without limitation, abdominal distension, chest pain, dizziness, heart palpitations, shortness of breath (dyspnea), swelling and fluid retention (edema) in the legs, heart failure, reduction in left ventricular ejection fraction, asymptomatic QT prolongation, and acute coronary syndromes (ACS). In some cases, the methods and materials described herein can be effective to delay the progression of one or more symptoms of cardiotoxicity in a mammal being administered one or more anti-cancer agents (e.g., one or more TKIs such as sunitinib) by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent. In some cases, the methods and materials described herein can be effective to prevent the onset of one or more symptoms of cardiotoxicity in a mammal being administered one or more anti-cancer agents (e g., one or more TKIs such as sunitinib) for, for example, from about 1 month to about 5 years (e.g., from about 1 month to about 4 years, from about 1 month to about 3 years, from about 1 month to about 2 years, from about 1 month to about 1 year, from about 1 month to about 6 months, from about 6 months to about 5 years, from about 1 year to about 5 years, from about 2 years to about 5 years, from about 3 years to about 5 years, from about 4 years to about 5 years, from about 6 months to about 5 years, from about 1 year to about 4 years, from about 2 years to about 3 years, from about 6 months to about 2 years, from about 1 year to about 3 years, or from about 2 years to about 4 years).

[0086] In some cases, one or more (e.g., one, two, three, four, or more) NPA7 polypeptides provided herein (e.g., one or more polypeptides (e.g., one or more chimeric polypeptides) that each include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein can be used to delay the progression of or prevent the onset of hypertension induced by one or more anti-cancer agents (e.g., TKI-induced cardiotoxicity (e.g., sunitinib-induced cardiotoxicity). For example, one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) can be administered to a mammal (e.g., a human) in need thereof (e.g., a human being administered of scheduled to be administered one or more anti-cancer agents) to delay the progression of or prevent the onset of hypertension. For Attorney Docket No. 07039-2323WO1

[0087] 2024-157 example, one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) can be administered to a mammal (e.g., a human) in need thereof (e.g., a human being administered of scheduled to be administered one or more anti-cancer agents) to reduce the blood pressure within the mammal. In some cases, the methods and materials described herein can be effective to delay the progression of hypertension in a mammal being administered one or more anticancer agents (e.g.. one or more TKIs such as sunitinib) by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent. In some cases, the methods and materials described herein can be effective to prevent the onset of hypertension in a mammal being administered one or more anti-cancer agents (e.g., one or more TKIs such as sunitinib) for, for example, from about 1 month to about 5 years (e.g., from about 1 month to about 4 years, from about 1 month to about 3 years, from about 1 month to about 2 years, from about 1 month to about 1 year, from about 1 month to about 6 months, from about 6 months to about 5 years, from about 1 year to about 5 years, from about 2 years to about 5 years, from about 3 years to about 5 years, from about 4 years to about 5 years, from about 6 months to about 5 years, from about 1 year to about 4 years, from about 2 years to about 3 years, from about 6 months to about 2 years, from about 1 year to about 3 years, or from about 2 years to about 4 years). In some cases, the methods and materials described herein can be effective to maintain a blood pressure in a mammal being administered one or more anti-cancer agents (e.g., one or more TKIs such as sunitinib) of less than about 140 / 90 millimeters of mercury (mm Hg).

[0088] An effective amount of a composition (e g., a pharmaceutical composition) containing one or more NPA7 polypeptides provided herein (e.g., one or more polypeptides (e.g., one or more chimeric polypeptides) that each include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein can be any amount that can treat a mammal (e.g., a human) as described herein (e.g., to treat or prevent cardiotoxicity and / or hypertension induced by one or more anti-cancer agents) without producing significant toxicity to the mammal. For example, an effective amount of one or more NPA7 polypeptides provided herein can be from about 100 picomoles of polypeptide(s) per kilogram body weight of the mammal (pmol / kg) per dose to about 1200 pmol / kg (e.g., from about 100 to about 1000, from about 100 to about 750, from about 100 to about 500, Attorney Docket No. 07039-2323WO1

[0089] 2024-157 from about 100 to about 250, from about 250 to about 1200, from about 500 to about 1200. from about 750 to about 1200, from about 1000 to about 1200, from about 200 to about 1000, from about 300 to about 900, from about 400 to about 800. from about 500 to about 700, from about 100 to about 300, from about 200 to about 400, from about 300 to about 500, from about 400 to about 600, from about 500 to about 700, from about 600 to about 800, from about 700 to about 900, or from about 800 to about 100 pmol / kg). The effective amount can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammafs response to treatment. Various factors can influence the actual effective amount used for a particular application. For example, the frequency of administration, duration of treatment, use of multiple treatment agents, route of administration, and severity of the condition may require an increase or decrease in the actual effective amount administered.

[0090] The frequency of administration of a composition (e.g., a pharmaceutical composition) containing one or more one or more NPA7 polypeptides provided herein (e.g., one or more polypeptides (e.g., one or more chimeric polypeptides) that each include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein can be any frequency that can treat a mammal (e.g., a human) as described herein (e.g., to treat or prevent cardiotoxicity and / or hypertension induced by one or more anti-cancer agents) without producing significant toxicity to the mammal. For example, the frequency of administration can be from about two times a day to about once a month, from about once a day to about tw ice a month, or from about once a week to about every two weeks. The frequency of administration can remain constant or can be variable during the duration of treatment. A course of treatment with a composition containing one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) can include rest periods. As with the effective amount, various factors can influence the actual frequency of administration used for a particular application. For example, the effective amount, duration of treatment, use of multiple treatment agents, route of administration, and severity of the condition may require an increase or decrease in administration frequency.

[0091] An effective duration for administering a composition (e.g., a pharmaceutical composition) containing one or more one or more NPA7 polypeptides provided herein Attorney Docket No. 07039-2323WO1

[0092] 2024-157

[0093] (e.g., one or more polypeptides (e.g., one or more chimeric polypeptides) that each include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein can be any duration that can treat a mammal (e.g., a human) as described herein (e.g., to treat or prevent cardiotoxicity and / or hypertension induced by one or more anti -cancer agents) without producing significant toxicity to the mammal. For example, the effective duration can vary from several days to several weeks, months, or years. In some cases, an effective duration for the treatment of a mammal (e.g., a human) having, or at risk of developing, cardiotoxicity and / or hypertension induced by one or more anti-cancer agents (e.g., TKI- induced cardiotoxicity (e.g., sunitinib-induced cardiotoxicity) and / or TKI-induced hypertension (e.g., sunitinib-induced hypertension)) can be the duration of the life of the mammal. Multiple factors can influence the actual effective duration used for a particular treatment. For example, an effective duration can vary7with the frequency of administration, effective amount, use of multiple treatment agents, route of administration, and severity of the condition being treated.

[0094] In some cases, one or more NPA7 polypeptides provided herein (e.g., one or more polypeptides (e.g., one or more chimeric polypeptides) that each include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein can be used as the sole active agent(s) to treat or prevent cardiotoxicity and / or hypertension in a mammal (e.g., a human) currently being administered or scheduled to be administered one or more anticancer agents (e g., one or more TKIs such as sunitinib).

[0095] In some cases, one or more NPA7 polypeptides provided herein (e.g., one or more polypeptides (e.g., one or more chimeric polypeptides) that each include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein can be used together with one or more (e.g., one, two, three, four, or more) additional agents that can treat cardiotoxicity and / or hypertension. In some cases, an agent that can be used to treat cardiotoxicity (or to treat one or more symptoms of cardiotoxicity) can be a beta blocker. In some cases, an agent that can be used to treat cardiotoxicity (or to treat one or more symptoms of cardiotoxicity) can be an angiotensin-converting enzyme (ACE) inhibitor. In some cases, an agent that can be used to treat cardiotoxicity (or to treat one or more symptoms of Attorney Docket No. 07039-2323WO1

[0096] 2024-157 cardiotoxicity) can be a diuretic. In some cases, an agent that can be used to treat cardiotoxicity (or to treat one or more symptoms of cardiotoxicity) can be a statin. Examples of agents that can be used to treat cardiotoxicity’ (or to treat one or more symptoms of cardiotoxicity) include, without limitation, aspirin, lisinopril (e.g., ZESTRIL®), fosinopril sodium (e.g., MONOPRIL®), metoprolol (e.g., LOPRESSOR®), atenolol (e.g., TENORMIN®), digitalis (e.g., LANOXIN®), isosorbide dinitrate (e.g., ISORDIL®), atorvastatin (e g., LIPITOR®), dexrazoxane (ZINECARD®), sacubitril / valsartan (ENTRESTO®), and any combinations thereof. In cases where one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) are used in combination with one or more additional agents used to treat cardiotoxicity (or to treat one or more symptoms of cardiotoxicity), the one or more additional agents can be administered at the same time (e.g., in a single composition containing one or more NPA7 polypeptides provided herein and the one or more additional agents) or independently. For example, one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) can be administered first, and the one or more additional agents administered second, or vice versa.

[0097] In some cases, an agent that can be used to treat hypertension can be a diuretic (e.g., a thiazide-like diuretic, a loop diuretic, or a potassium-sparing diuretic). In some cases, an agent that can be used to treat hypertension can be an ACE inhibitor. In some cases, an agent that can be used to treat hypertension can be an angiotensin II receptor blocker (ARB). In some cases, an agent that can be used to treat hypertension can be a calcium channel blocker. In some cases, an agent that can be used to treat hypertension can be a thiazide. In some cases, an agent that can be used to treat hypertension can be a calcium channel blocker (CCB). Examples of agents that can be used to treat hypertension include, without limitation, chlorthalidone, hydrochlorothiazide (e.g., MICROZIDE®), lisinopril (e.g., PRINIVIL® and ZESTRIL®), benazepril (e g., LOTENSIN®), captopril, candesartan (e.g., ATACAND®), losartan (e.g., COZ AAR®) amlodipine (e.g., NORVASC®), diltiazem (e.g., CARDIZEM® and TIAZAC®), furosemide, atenolol (e.g., TENORMIN®), indapamide. metolazone, ethacrynic acid, torsemide, triamterene, amiloride, spironolactone, enalapril, fosinopril, moexipril, perindopril, quinapril, ramipril, trandolapril, eprosartan, irbesartan, olmesartan, Attorney Docket No. 07039-2323WO1

[0098] 2024-157 telmisartan, valsartan, felodipine, lercanidipine, nicardipine, nifedipine, nisoldipine, verapamil, bisoprolol, carvedilol, labetalol, metoprolol (e.g., LOPRESSOR®), nebivolol, propranolol, and any combinations thereof. In cases where one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) are used in combination with one or more additional agents used to treat hypertension, the one or more additional agents can be administered at the same time (e.g., in a single composition containing one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) and the one or more additional agents) or independently. For example, one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) can be administered first, and the one or more additional agents administered second, or vice versa. Examples of therapies that can be used to treat hypertension include, without limitation, low-sodium diets, physical exercise, stress management, reduce / quit smoking and alcohol, weight loss, and ultrasound and radiofrequency renal denervation. In cases where one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) are used in combination with one or more additional therapies used to treat hypertension, the one or more additional therapies can be performed at the same time or independently of the administration of one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein). For example, one or more NPA7 polypeptides provided herein (and / or nucleic acids designed to express one or more NPA7 polypeptides provided herein) can be administered before, during, or after the one or more additional therapies are performed.

[0099] Exemplary Embodiments

[0100] Embodiment 1 is a method for delaying or preventing cardiotoxicity' in a mammal having been administered an anti-cancer agent, wherein said method comprises administering a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide, a nucleic acid encoding said polypeptide, or a composition comprising said polypeptide, said nucleic acid, or both to said mammal. Attorney Docket No. 07039-2323WO1

[0101] 2024-157

[0102] Embodiment 2 is a method for delaying or preventing cardiotoxicity' in a mammal scheduled to receive an anti-cancer agent, wherein said method comprises administering a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide, a nucleic acid encoding said polypeptide, or a composition comprising said polypeptide, said nucleic acid, or both to said mammal.

[0103] Embodiment 3 is the method of any one of embodiments 1-2, wherein said mammal is a human.

[0104] Embodiment 4 is the method of any one of embodiments 1-3, wherein said anticancer agent is a ty rosine kinase inhibitor (TKI).

[0105] Embodiment 5 is the method of any one of embodiments 1-3, wherein said anticancer agent is selected from the group consisting of sunitinib, sorafenib, dasatinib, lenvatinib, imatinib, nilotinib, bosutinib. and ponatinib.

[0106] Embodiment 6 is the method of any one of embodiments 1-5, wherein said natriuretic peptide is selected from the group consisting of an atrial natriuretic peptide (ANP), a brain natriuretic peptide (BNP), a C-type natriuretic peptide (CNP), a urodilatin (URO) peptide, and a Dendroaspis natriuretic peptide (DNP).

[0107] Embodiment 7 is the method of any one of embodiments 1-5, wherein said natriuretic peptide comprises, consists of, or consists essentially of any one of SEQ ID NOs:2-6.

[0108] Embodiment 8 is the method of any one of embodiments 1-7, wherein said ANG1- 7 peptide comprises, consists of. or consists essentially of the amino acid sequence set forth in SEQ ID NO: 7.

[0109] Embodiment 9 is the method of any one of embodiments 1-5, wherein said polypeptide comprises, consists of, or consists essentially of the amino acid sequence set forth in SEQ ID NO: I.

[0110] Embodiment 10 is the method of any one of embodiments 1-9, wherein said administering is subcutaneous, intravenous, or oral administration.

[0111] Embodiment 11 is a method for delaying or preventing hypertension in a mammal having been administered an anti-cancer agent, wherein said method comprises administering a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide, a nucleic acid encoding said polypeptide, or a composition comprising said polypeptide, said nucleic acid, or both to said mammal. Attorney Docket No. 07039-2323WO1

[0112] 2024-157

[0113] Embodiment 12 is a method for delaying or preventing hypertension in a mammal scheduled to receive an anti-cancer agent, wherein said method comprises administering a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide, a nucleic acid encoding said polypeptide, or a composition comprising said polypeptide, said nucleic acid, or both to said mammal.

[0114] Embodiment 13 is the method of any one of embodiments 11-12, wherein said mammal is a human.

[0115] Embodiment 14 is the method of any one of embodiments 11-13, wherein said anti-cancer agent is a TKI.

[0116] Embodiment 15 is the method of any one of embodiments 11-13, wherein said anti-cancer agent is selected from the group consisting of sunitinib, sorafenib, dasatinib, lenvatinib, imatinib, nilotinib, bosutinib. and ponatinib.

[0117] Embodiment 16 is the method of any one of embodiments 11-15, wherein said natriuretic peptide is selected from the group consisting of an ANP, a BNP, a CNP, a URO peptide, and a DNP.

[0118] Embodiment 17 is the method of any one of embodiments 11-15, wherein said natriuretic peptide comprises, consists of, or consists essentially of any one of SEQ ID NOs:2-6.

[0119] Embodiment 18 is the method of any one of embodiments 11-17, wherein said ANG1-7 peptide comprises, consists of, or consists essentially of the amino acid sequence set forth in SEQ ID NO: 7.

[0120] Embodiment 19 is the method of any one of embodiments 1 1-15, wherein said polypeptide comprises, consists of, or consists essentially of the amino acid sequence set forth in SEQ ID NO: 1.

[0121] Embodiment 20 is the method of any one of embodiments 11-19, wherein said administering is subcutaneous, intravenous, or oral administration.

[0122] Embodiment 21 is a use of a composition comprising a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide or a nucleic acid encoding said polypeptide to treat or prevent cardiotoxicity induced by an anticancer agent. Attorney Docket No. 07039-2323WO1

[0123] 2024-157

[0124] Embodiment 22 is a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide for use in the preparation of a medicament to treat or prevent cardiotoxicity induced by an anti-cancer agent.

[0125] Embodiment 23 is a nucleic acid encoding a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide for use in the preparation of a medicament to treat or prevent cardiotoxicity induced by an anti-cancer agent.

[0126] Embodiment 24 is a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide for use in the treatment of a medicament to treat or prevent cardiotoxicity induced by an anti -cancer agent.

[0127] Embodiment 25 is a nucleic acid encoding a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide for use in the treatment of a medicament to treat or prevent cardiotoxicity induced by an anti-cancer agent.

[0128] Embodiment 26 is a use of a composition comprising a polypeptide that includes (a) a natriuretic peptide and (b) anANGl-7 peptide or a nucleic acid encoding said polypeptide to treat or prevent hypertension induced by an anti-cancer agent.

[0129] Embodiment 27 is a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide for use in the preparation of a medicament to treat or prevent hypertension induced by an anti-cancer agent.

[0130] Embodiment 28 is a nucleic acid encoding a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide for use in the preparation of a medicament to treat or prevent hypertension induced by an anti-cancer agent.

[0131] Embodiment 29 is a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide for use in the treatment of a medicament to treat or prevent hypertension induced by an anti-cancer agent.

[0132] Embodiment 30 is a nucleic acid encoding a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide for use in the treatment of a medicament to treat or prevent hypertension induced by an anti-cancer agent.

[0133] The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims. Attorney Docket No. 07039-2323WO1

[0134] 2024-157

[0135] EXAMPLES

[0136] Example 1: NPA7 Polypeptides and TKI-induced Cardiotoxicity’

[0137] This Example describes the use of NPA7 polypeptides (e.g.. chimeric polypeptides that each include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides) in treating or preventing TKI-induced cardiotoxicity. A schematic showing an exemplary mechanism of NPA7 polypeptides in treating or preventing cardiotoxicity is shown in FIG. 1.

[0138] Methods

[0139] Bispecific NPA7 polypeptides w ere generated by fusing amino acids of a human BNP polypeptide with an ANG 1-7 polypeptide. A representative NPA7 polypeptide structure is shown in FIG. 2. NPA7 possessed enhanced natriuretic, diuretic, systemic, and renal vasorelaxant and cardiac unloading properties compared to BNP and ANG 1-7 alone (Meems et al., Hypertension 73:900-909 (2019)).

[0140] In vivo study: Male Wistar Kyoto rats (250-300 g), obtained from Charles River, were housed in individual cages and maintained on a 12-hour light / dark cycle, having access to standard laboratory rat chow and water ad libitum. Systolic BP and heart rate were measured by the CODA noninvasive BP system (a tail-cuff Method, Kent Scientific Corporation). Male WKY rats were randomly administered sunitinib (28 mg / kg per day) or vehicle by daily oral gavage (0.5 mL).

[0141] Western blotting: For western blotting, total protein extraction was performed using RIPA Lysis buffer (Millipore) supplemented with protease and phosphatase inhibitor cocktails (Roche), while nuclear-cytoplasmatic extraction w as accomplished using the NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific). Protein concentrations were determined with the BCA protein assay Kit (Thermo Fisher Scientific). Equal amounts of proteins (20 pg for the total lysate, 10 pg for the cytosolic fraction, and 5 pg for the nuclear fraction) were separated using 4-20% SDS-polyacrylamide gel electrophoresis, then transferred to PVDF membranes (Millipore). Membranes were blocked in 5% non-fat milk in IX Tris-Buffered Saline, 0.1% Tween 20 Detergent (TBST) for 1 hour and then incubated overnight with primary antibodies diluted in 5% non-fat milk in TBST. The target proteins were immunoblotted with the specific antibodies. Chemiluminescent substrate (Millipore) was used for detecting the signaling intensity. The following primary antibodies were used: anti-MFNl Attorney Docket No. 07039-2323WO1

[0142] 2024-157

[0143] (Cat#14793, Cell Signaling), anti-MFF (Cat#84580, Cell Signaling), anti-KEAPl (Cat# 10503-2 -AP, Proteintech), anti-NRF2 (Cat#16396-l-AP, Proteintech) at 1:1000 dilution.

[0144] RNA extraction. cDNA synthesis, and real-time quantitative PCR. Total RNA was extracted from HCMs using the TRIzol Reagent (Invitrogen) according to the manufacturer’s instructions. cDNA was synthesized by using Superscript III First-Strand Synthesis System (Thermo Fisher Scientific, Waltham, MA, USA) using 1 pg mRNA. Real-time reverse transcription PCR assays were conducted in 96-well plates using the LightCycler 480 Instrument (Roche, Wilmington, MA, USA). Levels of mRNA expression were normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). PCR primers used are listed in TABLE 2.

[0145] Immunofluorescence microscopy: For DHE staining in HCMs, cells were grown on glass coverslips and added 5 pM dihydroethidium working solution, and then incubated at room temperature for 30 minutes. For DHE staining in tissue, section slides were washed in PBS for 30 seconds and immediately placed in 5 pM DHE staining solution. The slides were incubated for 5-20 minutes at room temperature and avoided exposure to light. After DHE staining, the cells or the slides were washed twice with PBS and then detected by fluorescence microscope. For MitoTracker™ dye staining, cells were incubated in the labeling medium (100 nM) for 30 min at 37°C. Then, the labeling medium were aspirated, and cells were rinsed three times with culture medium. For the immunofluorescence of TOM20 or phalloidin, cells were fixed with 4% paraformaldehyde for 20 minutes at room temperature, followed by permeabilization with 0.1% Triton X-100 for 10 minutes. Cells were then blocked with 3% BSA and immune- stained with TOM20 antibody or the phalloidin solution (100 nM). Fluorescence images were acquired using Nikon TE2000-U with Metamorph software (Molecular Devices).

[0146] Measurement of ATP levels: Intracellular ATP concentration was assessed using the ATP Assay Kit-Luminescence (Cayman Chemical; Ann Arbor, MI). Briefly, HCMs were seeded into a 96-well plate and grown to 70% confluence. Cells were then pretreated with NPA7 for 6 hours, followed by incubation with sunitinib (10 pM) for 24 hours. After treatment, HCMs were homogenized in lysis buffer and centrifuged at 12,000 g for 5 minutes at 4°C to obtain cell-free supernatants. For ATP detection, 100 pL of ATP detection w orking solution w as added to each well and incubated at room Attorney Docket No. 07039-2323WO1

[0147] 2024-157 temperature for 5 minutes to deplete background ATP. Subsequently, 10 pL of either the samples or standards were added to the wells. Luminescence was measured using a microplate reader after a 15-minute incubation at room temperature.

[0148] Measurements ofMMP: MMP was measured using tetramethylrhodamine methyl ester (TMRM) fluorescence imaging with the TMRM Assay Kit (Abeam, ab228569), following the manufacturer’s instructions. Briefly, HCMs were seeded into 6-well plate and grown to 70% confluence. Cells were then pretreated with NPA7 for 6 hours, followed by incubation with sunitinib (10 pM) for 24 hours. After treatment, HCMs were incubated with 100 nM TMRM reagent diluted serum-free medium at 37°C for 20 minutes in the dark. Fluorescent images were acquired using a fluorescence microscope at an excitation / emission wavelength of 548 / 573 nm. The fluorescence intensity was quantified using ImageJ software.

[0149] LDH cytotoxicity assay: Cytotoxicity was evaluated by measuring LDH release using the LDH Cytotoxicity Assay Kit (MCE, HY-K1090), following the manufacturer’s instructions. LDH activity was quantified by a coupled enzymatic reaction in which LDH catalyzed the conversion of lactate to pyruvate, accompanied by the reduction of NAD+. The resulting formation of formazan was directly proportional to the amount of LDH released into the medium, which was indicative of cytotoxicity. Briefly, HCMs were seeded into 6-well plate and grown to 70% confluence. Cells were pretreated with NPA7 for 6 hours, followed by incubation with sunitinib (10 pM) for 24 hours. After treatment, 50 pL of sample medium was transferred into triplicate wells of a 96-well flat bottom plate. For the LDH positive control, 50 pL of LDH Positive Control reagent was added to separate triplicate wells. Next, 50 pL of reaction mixture was added to each sample well, gently mixed and incubated at 37°C for 30 minutes in the dark. The reaction was stopped by adding Stop Solution, and absorbance was measured at 490 nm using a microplate reader.

[0150] Measurements of mitochondrial ROS: Mitochondrial ROS were detected using MitoSOX Red reagent (Invitrogen) at a final concentration of 5 pM. HCMs w ere incubated with MitoSOX at 37°C for 10 minutes in the dark. Following incubation, cells were washed with PBS for 5 minutes. Fluorescence images were acquired using a Leica DM2000 fluorescence microscopy, and quantification of fluorescence intensity was performed using ImageJ software. Attorney Docket No. 07039-2323WO1 2024-157

[0151] GC-A andMasR activation in HCMs: For HCMs, 5 x 105ofHCMs / well were grown in 6-well plates until 80 to 90% confluency and were then treated with PBS or NPA7 at doses of 109. 10s. or 106M for 10 minutes at 37 °C. Afterward, cells were washed with PBS and lysed, sonicated, centrifuged, and the supernatants were extracted and reconstituted in 300 pL O. I M HC1 for cGMP or cAMP assay. The samples were measured using a commercial cGMP or cAMP ELISA kit (Enzo Life Sciences) as instructed by the manufacturer.

[0152] TABLE 2. Specific primer pairs used in this study.

[0153] Results

[0154] NPA7 generated the second messengers cGMP and cAMP in vitro via GC-A and MasR, respectively. Bioengineered NPA7 simultaneously co-activated the protective GC- A / cGMP and MasR / cAMP pathways. As illustrated in FIGS. 3A and 3B, NPA7 significantly increased the concentrations of cGMP and cAMP in HCMs compared to no treatment.

[0155] NPA7 reduced blood pressure in spontaneously hypertensive rats (SHRs). During a continuous infusion time course study, dosing SHRs with 300, 600 and 1200 pmol / kg NPA7 (n=4) significantly decreased systolic BP when compared to SHRs that received Attorney Docket No. 07039-2323WO1

[0156] 2024-157

[0157] NPA7 promoted the release of nitric oxide (NO) in endothelial cells. NPA7 significantly increased total nitrite / nitrate level, which represents higher NO release in cardiac endothelial cells (FIG. 5).

[0158] NPA7 acted as a NRF2 activator which protects against oxidative stress via the KEAP1-NRF2 pathway in HCMs. Dihydroethidium (DHE) is a fluorescent probe that is used for the detection of reactive oxygen species (ROS) levels. In FIGS. 6A and 6B, it is demonstrated that H2O2 increased DHE staining and that treatment with NPA7 attenuated H2O2 induced ROS upregulation. Meanwhile, NPA7 increased the mRNA levels of antioxidant genes (G6PD, GSS, and GCLC) after H2O2 exposure (FIG. 6C). NPA7 enhanced the NRF2 protein expression level and KEAP1 degradation (FIG. 6D).

[0159] Sunitinib induced oxidative stress in HCMs. Increased oxidative stress contributes to cardiotoxicity and the rise in BP during VEGF inhibition induced by sunitinib. Sunitinib treatment increased DHE staining intensity using ROS as an indicator (FIGS. 7A and 7B).

[0160] Sunitinib induced oxidative stress-dependent mitochondrial fragmentation and hypertrophy in HCMs. Mitochondrial morphology was determined by immunofluorescence of Tom20. HCMs treated with vehicle displayed rod-like networked mitochondria, while HCMs treated with sunitinib exhibited circular and punctate mitochondria, indicating mitochondrial fragmentation (FIG. 8). The role of oxidative stress in sunitinib-induced mitochondrial fragmentation was examined. Treatment with 5 mM N-acetyl-L-cysteine (NAC), a ROS inhibitor, rescued mitochondria from sunitinib- induced fragmentation (FIG. 8). Sunitinib treatment increased the cell surface area and hypertrophic genes (NPPA an< MYPI7) in HCMs, whereas NAC abolished these markers (FIG. 9).

[0161] Sunitinib increased BP in WKY rats. Male WKY rats were randomly administered sunitinib (28 mg / kg per day; n=3) or vehicle (n=3) by oral gavage (0.5 mL) for 6 days. Systolic BP was measured by the CODA noninvasive BP system (a tail-cuff Method, Kent Scientific Corporation). Daily administration of sunitinib to WKY rats induced a rise in SBP at Day 6 (FIG. 10A). In addition, sunitinib treatment significantly increased the hypertrophic gene markers (Nppa and Nppb) in the left ventricle of WKY rats (FIG. 10B). Attorney Docket No. 07039-2323WO1

[0162] 2024-157

[0163] Sunitinib treatment increased the cell surface area and hypertrophic genes (NPPA) in HCMs, whereas NPA7 treatment abolished these markers (FIG. 11).

[0164] Mitochondrial morphology was determined by immunofluorescence of MitoTracker™ dye staining. HCMs treated with vehicle displayed rod-like networked mitochondria, while HCMs treated with sunitinib exhibited circular and punctate mitochondria, indicating mitochondrial fragmentation (FIG. 12). Treatment with 10 uM NP7A polypeptides inhibited sunitinib-induced mitochondrial fragmentation of HCMs (FIG. 12).

[0165] Consistent with the results of Mitochondrial morphology, sunitinib induced reduction of mitofusin-1 fission (MFN1) polypeptides and increase of mitochondrial fission factor (MFF) polypeptide levels in HCMs, whereas NPA7 attenuated these effects (FIG. 13). As shown in FIG. 14, NPA7 polypeptides promoted KEAP1 degradation and NRF2 activation in HCMs treated with sunitinib.

[0166] In addition, NPA7 inhibited sunitinib-induced LDH cytotoxicity' in HCMs (FIG. 15), and rescued the reduction in ATP levels induced by sunitinib in a dose-dependent manner in HCMs (FIG. 16). NPA7 also rescued the reduction in MMP induced by sunitinib in a dose-dependent manner in HCMs, as indicated by the immunofluorescent images shown in FIG. 17A and the intensity' quantification plotted in FIG. 17B for TMRM staining. Further, NPA7 inhibited sunitinib-induced production of mitochondrial ROS in HCMs. as demonstrated by immunofluorescent images (FIG. 18 A) and intensity quantification (FIG. 18B) for MitoSox staining.

[0167] Example 2: Preventing TKI-Induced Cardiotoxicity

[0168] A human having cancer and being administered or scheduled to be administered one or more anti-cancer agents (e.g.. one or more TKIs such as sunitinib) is administered one or more NPA7 polypeptides provided herein (e.g., one or more polypeptides (e.g., one or more chimeric polypeptides) that each include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides). In some cases, the administered NPA7 polypeptides can prevent the onset of one or more symptoms of cardiotoxicity induced by the anticancer agent(s). In some cases, the administered NPA7 polypeptides can delay the progression of one or more symptoms of cardiotoxicity induced by the anti -cancer agent(s). Attorney Docket No. 07039-2323WO1 2024-157

[0169] Example 3: Preventing TKI-Induced Hypertension

[0170] A human having cancer and being administered or scheduled to be administered one or more anti-cancer agents (e.g.. one or more TKIs such as sunitinib) is administered one or more NPA7 polypeptides provided herein (e.g., one or more polypeptides (e.g., one or more chimeric polypeptides) that each include (a) one or more natriuretic peptides and (b) one or more ANG1-7 peptides). In some cases, the administered NPA7 polypeptides can prevent hypertension induced by the anti-cancer agent(s). In some cases, the administered NPA7 polypeptides can delay the onset of hypertension induced by the anticancer agent(s). In some cases, the administered NPA7 polypeptides can maintain human’s blood pressure at less than about 140 / 90 mm Hg.

[0171] OTHER EMBODIMENTS

[0172] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

Attorney Docket No. 07039-2323WO12024-157WHAT IS CLAIMED IS:

1. A method for delaying or preventing cardiotoxicity in a mammal having been administered an anti-cancer agent, wherein said method comprises administering a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide, a nucleic acid encoding said polypeptide, or a composition comprising said polypeptide, said nucleic acid, or both to said mammal.

2. A method for delaying or preventing cardiotoxicity in a mammal scheduled to receive an anti -cancer agent, wherein said method comprises administering a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide, a nucleic acid encoding said polypeptide, or a composition comprising said polypeptide, said nucleic acid, or both to said mammal.

3. The method of claim 1 or claim 2, wherein said mammal is a human.

4. The method of claim 1 or claim 2, wherein said anti-cancer agent is a tyrosine kinase inhibitor (TKI).

5. The method of claim 1 or claim 2, wherein said anti-cancer agent is selected from the group consisting of sunitinib, sorafenib, dasatinib, lenvatinib, imatinib, nilotinib, bosutinib, and ponatinib.

6. The method of claim 1 or claim 2, wherein said natriuretic peptide is selected from the group consisting of an atrial natriuretic peptide (ANP), a brain natriuretic peptide (BNP), a C-type natriuretic peptide (CNP), a urodilatin (URO) peptide, and a Dendroaspis natriuretic peptide (DNP).

7. The method of claim 1 or claim 2, wherein said natriuretic peptide comprises, consists of, or consists essentially of any one of SEQ ID NOs:2-6.

8. The method of claim 1 or claim 2, wherein said ANG1-7 peptide comprises, consists of, or consists essentially of the amino acid sequence set forth in SEQ ID NO:7.

9. The method of claim 1 or claim 2, wherein said polypeptide comprises, consists of, or consists essentially of the amino acid sequence set forth in SEQ ID NO: 1.Attorney Docket No. 07039-2323WO12024-15710. The method of claim 1 or claim 2, wherein said administering is subcutaneous, intravenous, or oral administration.

11. A method for delaying or preventing hypertension in a mammal having been administered an anti-cancer agent, wherein said method comprises administering a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide, a nucleic acid encoding said polypeptide, or a composition comprising said polypeptide, said nucleic acid, or both to said mammal.

12. A method for delaying or preventing hypertension in a mammal scheduled to receive an anti-cancer agent, wherein said method comprises administering a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide, a nucleic acid encoding said polypeptide, or a composition comprising said polypeptide, said nucleic acid, or both to said mammal.

13. The method of claim 11 or claim 12. wherein said mammal is a human.

14. The method of claim 11 or claim 12, wherein said anti-cancer agent is a TKI.

15. The method of claim 11 or claim 12, wherein said anti-cancer agent is selected from the group consisting of sunitinib, sorafenib, dasatinib, lenvatinib, imatinib, nilotinib, bosutinib, and ponatinib.

16. The method of claim 11 or claim 12. wherein said natriuretic peptide is selected from the group consisting of an ANP, a BNP, a CNP, a URO peptide, and a DNP17. The method of claim 11 or claim 12, wherein said natriuretic peptide comprises, consists of. or consists essentially of any one of SEQ ID NOs:2-6.

18. The method of claim 11 or claim 12, wherein said ANG1-7 peptide comprises, consists of, or consists essentially of the amino acid sequence set forth in SEQ ID NO: 7.

19. The method of claim 11 or claim 12, wherein said polypeptide comprises, consists of, or consists essentially of the amino acid sequence set forth in SEQ ID NO: 1.Attorney Docket No. 07039-2323WO12024-15720. The method of claim 11 or claim 12, wherein said administering is subcutaneous, intravenous, or oral administration.

21. Use of a composition comprising a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide or a nucleic acid encoding said polypeptide to treat or prevent cardiotoxicity induced by an anti-cancer agent.

22. A polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1 -7 peptide for use in the preparation of a medicament to treat or prevent cardio toxicity induced by an anti -cancer agent.

23. A nucleic acid encoding a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide for use in the preparation of a medicament to treat or prevent cardiotoxicity induced by an anti-cancer agent.

24. A polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1 -7 peptide for use in the treatment of a medicament to treat or prevent cardio toxicity induced by an anti-cancer agent.

25. A nucleic acid encoding a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide for use in the treatment of a medicament to treat or prevent cardiotoxicity induced by an anti -cancer agent.

26. Use of a composition comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide or a nucleic acid encoding said polypeptide to treat or prevent hypertension induced by an anti-cancer agent.

27. A polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1 -7 peptide for use in the preparation of a medicament to treat or prevent hypertension induced by an anti-cancer agent.

28. A nucleic acid encoding a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1 -7 peptide for use in the preparation of a medicament to treat or prevent hypertension induced by an anti-cancer agent.Attorney Docket No. 07039-2323WO1 2024-15729. A polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1 -7 peptide for use in the treatment of a medicament to treat or prevent hypertension induced by an anti-cancer agent.

30. A nucleic acid encoding a polypeptide comprising a polypeptide that includes (a) a natriuretic peptide and (b) an ANG1-7 peptide for use in the treatment of a medicament to treat or prevent hypertension induced by an anti-cancer agent.

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