Methods and compositions for prolonged renalase agonist activity

Abstract

Administration of recombinant or biologically-isolated renalase has been shown to treat certain diseases and conditions. Stable synthetic peptides exhibiting renalase agonist activity are highly desirable. Disclosed are compositions comprising modified renalase agonist peptides that display increased in vitro and / or in vivo half-life, and methods for their use in treating diseases such as ischemic and toxic organ injury.

Description

[0001] TITLE OF THE INVENTION

[0002] METHODS AND COMPOSITIONS FOR PROLONGED RENALASE AGONIST ACTIVITY

[0003] CROSS-REFERENCE TO RELATED APPLICATIONS

[0004] This application claims priority to U.S. Provisional Patent Application No. 63 / 518,160, filed August 8, 2023, which is hereby incorporated herein by reference in its entirety.

[0005] BACKGROUND OF THE INVENTION

[0006] Renalase (“RNLS”) is secreted by the kidney and has multiple biological functions. Administration of recombinant renalase or peptide fragments of renalase has been shown to treat certain diseases and disorders. Stable peptides exhibiting renalase agonist activity are highly desirable. There remains a need in the art for improved renalase peptides exhibiting renalase agonist activity. This invention satisfies this unmet need.

[0007] SUMMARY OF THE INVENTION

[0008] In various aspects, the present invention provides a peptide comprising at least one amino acid sequence that is at least 90% identical to the amino acid sequence selected from SEQ ID NOs: 5, 6, 21, or 26. In some embodiments, the peptide further comprises at least one N-terminal glycine residue. In some embodiments, the peptide further comprises at least one N-terminal glycine residue is linked to at least one molecule that enhances association of the peptide with human serum albumin. In some embodiments, the at least one molecule that enhances association of the peptide with human serum albumin is selected from myristoleic acid and palmitoleic acid. In some embodiments, the peptide is amidated at the C-terminus.

[0009] In some embodiments, the peptide is N-terminally linked to human serum albumin. In some embodiments, the human serum albumin is at least 90% identical to the amino acid sequence of SEQ ID NO: 17. In some embodiments, the peptide is N- terminally linked to human serum albumin through a linker. In some embodiments, the linker is at least 90% identical to SEQ ID NO: 2. In some embodiments, the linker is (G4S)2. In some embodiments, the peptide is C-terminally linked to a sequence that is at least 90% identical to the amino acid sequence set forth in SEQ ID NO: 8. In some embodiments, the peptide is C-terminally linked to a Strep tag. In some embodiments, the Strep Tag is at least 90% identical to the amino acid sequence of SEQ ID NO: 3.

[0010] In some embodiments, the peptide comprises a renalase agonist. In some embodiments, the half-life of the peptide is at least 3 days.

[0011] In some embodiments, the present invention provides a composition comprising the peptide. In some embodiments, the composition comprising the peptide is a pharmaceutical composition.

[0012] In some embodiments, the present invention provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the peptide or the pharmaceutical composition comprising the peptide.

[0013] In some embodiments, the present invention provides a modified renalase agonist peptide comprising at least one amino acid sequence that is at least 90% identical to SEQ ID NOs: 5, 6, 13, 14, 15, 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36.

[0014] In some embodiments, the modified renalase agonist peptide comprises SEQ ID NOs: 5, 6, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36, and further comprises at least one N-terminal glycine residue.

[0015] In some embodiments, the modified renalase agonist peptide comprises at least one modification selected from addition of a myristoleic acid residue or addition of a palmitoleic acid residue, and wherein the modified renalase agonist peptide is amidated at the C-terminus. In some embodiments, the addition of a myristoleic acid residue and addition of a palmitoleic acid residue is at the N’ terminus of the modified renalase agonist peptide.

[0016] In some embodiments, the modified renalase agonist comprises SEQ ID NO: 13, 14, 15, or 16.

[0017] In some embodiments, the modified renalase agonist peptide further comprises at least one modification to increase half-life or stability of the modified renalase agonist peptide. In some embodiments, the at least one modification is at least one D amino acid.

[0018] In some embodiments, the present invention provides a fusion peptide comprising a modified renalase agonist peptide, wherein the modified renalase agonist peptide is linked to an additional amino acid sequence or peptide. In some embodiments, the modified renalase agonist peptide is N-terminally linked to human serum albumin. In some embodiments, the human serum albumin is at least 90% identical to the amino acid sequence of SEQ ID NO: 17. In some embodiments, the fusion peptide comprises a linker between the modified renalase agonist peptide and the additional amino acid sequence or peptide.

[0019] In some embodiments, the half-life of the modified renalase agonist peptide or fusion peptide is increased as compared to an unmodified renalase peptide.

[0020] In some embodiments, the at least one N-terminal glycine residue of the modified renalase agonist peptide is linked to at least one molecule that enhances association of the peptide with human serum albumin.

[0021] In some embodiments, the present invention provides a composition comprising the modified renalase agonist peptide or fusion peptide. In some embodiments, the composition comprises at least one modified renalase agonist peptide and at least one fusion peptide, or at least two modified renalase agonist peptides, or at least two fusion peptides. In some embodiments, the composition is a pharmaceutical composition. In some embodiments, the composition further comprises at least one pharmacologically acceptable carrier.

[0022] In some embodiments, the present invention provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the modified renalase agonist peptide, fusion peptide, or composition of the present invention.

[0023] In some embodiments, the disease or disorder comprises a renal condition, a cardiovascular condition, a heart condition, a central nervous system (CNS) condition, pre-eclampsia. In some embodiments, the cardiovascular conditions comprises hypertension, asymptomatic left ventricular dysfunction, chronic congestive heart failure (CHF), myocardial infarction (MI), cardiac rhythm disturbance, stroke, atherosclerosis, and any combination thereof, the hypertension comprises chronic hypertension, systolic hypertension, isolated systolic hypertension, diabetic hypertension, pulmonary hypertension, acute severe hypertension, and any combination thereof, the renal condition comprises end-stage renal disease (ESRD), chronic renal failure, and any combination thereof, and the CNS condition comprises depression, anxiety, mania, schizophrenia, post-traumatic stress disorder (PTSD), and any combination thereof.

[0024] BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The following detailed description of embodiments of the invention will be better understood when read in conjunction with the appended drawings. It should be understood that the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings.

[0026] Figure 1, comprising Figure 1A and Figure IB, depicts representative illustrations of recombinant proteins used in this invention. Figure 1A depict a representative illustration of the predicted structure of mouse albumin (SEQ ID NO: 1). Figure IB depicts a representative schematic of a generic recombinant protein construct comprising a protein of interest, bearing a C-terminal Strep tag (SEQ ID NO: 3), linked to mouse albumin (SEQ ID NO: 1) via a linker peptide (SEQ ID NO: 2).

[0027] Figure 2 comprising Figure 2A through Figure 2C, depicts representative data demonstrating the purification of four recombinant proteins fused to mouse albumin. Figure 2A depicts representative schematics presenting four recombinant protein constructs. The recombinant human renalase construct (SEQ ID NO: 9) comprises full full-length human renalase (RNLS) (SEQ ID NO: 4) linked N-terminally to SEQ ID NO: 1 via a (G4S)2 linker (SEQ ID NO: 2) and C-terminally Strep tagged (SEQ ID NO: 3). The renalase peptide (RP) 81 recombinant construct (SEQ ID NO: 10) comprises RP81 (SEQ ID NO: 5) linked N-terminally to SEQ ID NO: 1 via a (G4S)2linker (SEQ ID NO:

[0028] 2), and bearing a C-terminal Strep tag (SEQ ID NO: 3). The RP10 recombinant construct (SEQ ID NO: 11) comprises RP10 (SEQ ID NO: 6) linked N-terminally to SEQ ID NO:

[0029] 1 via a (G4S)2linker (SEQ ID NO: 2), and bearing a C-terminal Strep tag (SEQ ID NO:

[0030] 3). The negative control recombinant construct (SEQ ID NO: 12) comprises a negative control 6-peptide (SEQ ID NO: 7) repeat sequence (SEQ ID NO: 8) linked N-terminally to SEQ ID NO: 1 via a (G4S)2 linker (SEQ ID NO: 2) and C-terminally Strep tagged (SEQ ID NO: 3). Figure 2B depicts a representative Coomassie blue stain demonstrating the production and purification of the four recombinant proteins depicted in Figure 2A. From left to right protein ladder, SEQ ID NO: 11, SEQ ID NO: 10, SEQ ID NO: 12, and SEQ ID NO: 9. Figure 2C depicts representative data presenting the protein 50ml culture yield of the recombinant proteins presented in Figure 2B.

[0031] Figure 3 depicts a representative schematic to conduct in vivo protein kinetic (PK) studies on recombinant protein SEQ ID NO: 11.

[0032] Figure 4 depicts a representative illustration of the enzyme-linked immunosorbent assay (ELISA) technique to quantitate recombinant proteins fused to mouse albumin.

[0033] Figure 5 depicts representative data presenting a standard curve used to determine protein concentration of recombinant protein SEQ ID NO: 11. y-axis: absorbance; x-axis: protein concentration (ng / ml).

[0034] Figure 6 depicts representative data presenting the stability of recombinant protein SEQ ID NO: 11 over time. Presented are representative data from three experiments (1-3). y-axis: absorbance at 450nm; x-axis: time (days).

[0035] Figure 7 depicts representative data demonstrating the increased in vivo half-life of recombinant protein SEQ ID NO: 11. Specifically, 20mg / kg of recombinant protein SEQ ID NO: 11 were intravenously (retro-orbital) injected into a mouse. Presence of recombinant protein SEQ ID NO: 11 was quantitated in extracted mouse plasma over time and quantitated via ELISA (Figure 4) to determine the half-life of SEQ ID NO: 11. y-axis: SEQ ID NO: 11 concentration (pg / ml); x-axis: time (days).

[0036] Figure 8 depicts representative data presenting the half-life of recombinant protein SEQ ID NO: 11. Presented are representative data from three experiments (1-3). y-axis: absorbance; x-axis: protein concentration.

[0037] Figure 9 depicts representative sequences of modified renalase agonist peptides. From top to bottom: The sequence of SEQ ID NO: 13 comprises SEQ ID NO: 5 with an additional N-terminal glycine residue conjugated to myristoleic acid and C- terminally amidated; the sequence of SEQ ID NO: 14 comprises SEQ ID NO: 5 with an additional N-terminal glycine residue conjugated to palmitoleic acid and C-terminally amidated; the sequence of SEQ ID NO: 15 comprises SEQ ID NO: 6 with an additional N-terminal glycine residue conjugated to myristoleic acid and C-terminally amidated; the sequence of SEQ ID NO: 16 comprises SEQ ID NO: 6 with an additional N-terminal glycine residue conjugated to palmitoleic acid and C-terminally amidated.

[0038] Figure 10 depicts representative in vitro cisplatin toxicity study data demonstrating the increased potency of the renalase peptide SEQ ID NO: 16 compared to renalase peptide SEQ ID NO: 6. y-axis: relative cell viability; x-axis: peptide concentration (pM).

[0039] DETAILED DESCRIPTION

[0040] The present invention is based, in part, on the unexpected findings that certain peptide fragments within the sequence of the human renalase protein exhibit renalase agonist activity and that specific modifications to the certain peptide fragments within the sequence of the human renalase protein that exhibit renalase agonist activity increases their half-life in vivo and / or in vitro.

[0041] As such, in one embodiment, the present invention provides at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro halflife. In one embodiment, the at least one modified renalase agonist peptide comprises at least one fragment of full-length human renalase protein (SEQ ID NO: 4), or a variant thereof. In one embodiment, the at least one modified renalase agonist peptide comprises at least one fragment of full-length human renalase protein (SEQ ID NO: 4), wherein the at least one fragment comprises an amino acid sequence that is at least 90% identical to a sequence of SEQ ID NOs: 5, 6, 21, or 26.

[0042] In one embodiment, the at least one modified renalase agonist peptide is modified to comprise at least one N-terminal glycine residue. In one embodiment, the at least one modified renalase agonist peptide is amidated at the C-terminus. In one embodiment, the at least one N-terminal glycine residue is linked to at least one molecule that enhances association of the at least one modified renalase agonist peptide to human serum albumin in vivo and / or in vitro. In one embodiment, at least one molecule that enhances association of the at least one modified renalase agonist peptide to human serum albumin in vivo and / or in vitro is selected from myristoleic acid and palmitoleic acid.

[0043] In another embodiment, the at least one modified renalase agonist peptide is N-terminally linked to human serum albumin or a fragment thereof. In one embodiment, the at least one modified renalase agonist peptide is N-terminally linked to human serum albumin or a fragment thereof, wherein the human serum albumin comprises a sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 17. In one embodiment, the at least one modified renalase agonist peptide is N- terminally linked to human serum albumin or a fragment thereof through a linker wherein the linker comprises (G4S)2 (SEQ ID NO: 2). In one embodiment, the at least one modified renalase agonist peptide is N-terminally linked to human serum albumin or a fragment thereof through a linker wherein the linker comprises a sequence that is at least 90% identical to SEQ ID NO: 2. In one embodiment, the at least one modified renalase agonist peptide is C-terminally linked to LEAS. In one embodiment, the at least one modified renalase agonist peptide is C-terminally linked to a Strep Tag (SEQ ID NO: 3). In one embodiment, the at least one modified renalase agonist peptide is C-terminally linked to a sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 3.

[0044] In one embodiment, the present invention provides at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life, wherein the increased in vivo and / or in vitro half-life comprises an in vivo and / or in vitro half-life of at least 1 day.

[0045] In one embodiment, the present invention provides a composition comprising at least one modified renalase agonist peptide, described herein, which displays increased in vivo and / or in vitro half-life wherein the composition is a pharmaceutical composition.

[0046] In one embodiment, the present invention provides a method of preventing or treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the at least one modified renalase peptide, described herein, which displays increased in vivo and / or in vitro half-life. In another embodiment, the present invention provides a method of preventing or treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising at least one modified renalase agonist peptide, described herein, which displays increased in vivo and / or in vitro half-life, and wherein the composition is a pharmaceutical composition.

[0047] In one embodiment, the present invention provides a method of preventing or treating a subject having, suspected of having, or at risk for having a disease or disorder. Exemplary diseases or disorders that are treatable or preventable by way of the compositions and methods of the present invention include, but are not limited to, ischemic and toxic organ injury. In various embodiments, the compositions and methods described herein can be used to prevent or treat any disease or disorder that is preventable or treatable by renalase agonist activity.

[0048] In some embodiments, the present invention provides a modified renalase agonist peptide comprising at least one amino acid sequence that is at least 90% identical to the amino acid sequence selected of SEQ ID NOs: 5, 6, 13, 14, 15, 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36. In some embodiments, the present invention provides a fusion peptide comprising a modified renalase agonist peptide, wherein the modified renalase agonist peptide is linked to an additional amino acid sequence or peptide. In some embodiments, the present invention provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the modified renalase agonist peptide, fusion peptide, or the pharmaceutical composition of the present invention.

[0049] Definitions

[0050] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although any methods and materials similar or equivalent to those described herein can be used in the practice for testing of the present invention, the preferred materials and methods are described herein. In describing and claiming the present invention, the following terminology will be used. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0051] The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0052] “About” as used herein when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass non-limiting variations of ±40% or ±20% or ±10%, ±5%, ±1%, or ±0.1% from the specified value, as such variations are appropriate.

[0053] The term “abnormal” when used in the context of organisms, tissues, cells or components thereof, refers to those organisms, tissues, cells or components thereof that differ in at least one observable or detectable characteristic (e.g., age, treatment, time of day, etc.) from those organisms, tissues, cells or components thereof that display the “normal” (expected) respective characteristic. Characteristics which are normal or expected for one cell or tissue type, might be abnormal for a different cell or tissue type.

[0054] The term “agonist” as used herein refers to a peptide, polypeptide, protein, a naturally occurring molecule or a synthetic molecule such as a drug, that binds to a receptor inside a cell or on its surface and causes the same or similar activity as the substance that normally binds to the receptor. As used herein, a renalase agonist causes the same or similar physiological response as the renalase protein when added to a biological system such as one or more cells in vitro, or an organism, including a human, in vivo.

[0055] As used herein, to “alleviate” or “treat” a disease means reducing the frequency or severity of at least one sign or symptom of a disease or disorder.

[0056] As used herein the terms “alteration,” “defect,” “variation,” or “mutation,” refers to a mutation in a gene in a cell that affects the function, activity, expression (transcription or translation) or conformation of the polypeptide that it encodes. Mutations encompassed by the present invention can be any mutation of a gene in a cell that results in the enhancement or disruption of the function, activity, expression or conformation of the encoded polypeptide, including the complete absence of expression of the encoded protein and can include, for example, missense and nonsense mutations, insertions, deletions, frameshifts and premature terminations. Without being so limited, mutations encompassed by the present invention may alter splicing the mRNA (splice site mutation) or cause a shift in the reading frame (frameshift).

[0057] By the term “applicator,” as the term is used herein, is meant any device including, but not limited to, a hypodermic syringe, a pipette, an iontophoresis device, a patch, and the like, for administering the compositions of the invention to a subject.

[0058] As used herein, the term “marker” or “biomarker” is meant to include a parameter which is useful according to this invention for determining the presence and / or severity of AKI.

[0059] The level of a marker or biomarker “significantly” differs from the level of the marker or biomarker in a reference sample if the level of the marker in a sample from the patient differs from the level in a sample from the reference subject by an amount greater than the standard error of the assay employed to assess the marker, and preferably at least 10%, and more preferably 25%, 50%, 75%, or 100%.

[0060] The term “coding sequence,” as used herein, means a sequence of a nucleic acid or its complement, or a part thereof, that can be transcribed and / or translated to produce the mRNA and / or the polypeptide or a fragment thereof. Coding sequences include exons in a genomic DNA or immature primary RNA transcripts, which are joined together by the cell's biochemical machinery to provide a mature mRNA. The anti-sense strand is the complement of such a nucleic acid, and the coding sequence can be deduced therefrom. In contrast, the term “non-coding sequence,” as used herein, means a sequence of a nucleic acid or its complement, or a part thereof, that is not translated into amino acid in vivo and / or in vitro, or where tRNA does not interact to place or attempt to place an amino acid. Non-coding sequences include both intron sequences in genomic DNA or immature primary RNA transcripts, and gene-associated sequences such as promoters, enhancers, silencers, and the like.

[0061] As used herein, the terms “complementary” or “complementarity” are used in reference to polynucleotides (i.e., a sequence of nucleotides) related by the basepairing rules. For example, the sequence “A-G-T,” is complementary to the sequence “T- C-A ” Complementarity may be “partial,” in which only some of the nucleic acids' bases are matched according to the base pairing rules. Or, there may be “complete” or “total” complementarity between the nucleic acids. The degree of complementarity between nucleic acid strands has significant effects on the efficiency and strength of hybridization between nucleic acid strands. This is of particular importance in amplification reactions, as well as detection methods that depend upon binding between nucleic acids.

[0062] A “disease” is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal’s health continues to deteriorate. In contrast, a “disorder” in an animal is a state of health in which the animal is able to maintain homeostasis, but in which the animal’s state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decrease in the animal’s state of health.

[0063] An “effective amount” as used herein, means an amount which provides a therapeutic or prophylactic benefit.

[0064] “Encoding” refers to the inherent property of specific sequences of nucleotides in a polynucleotide, such as a gene, a cDNA, or an mRNA, to serve as templates for synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (i.e., rRNA, tRNA and mRNA) or a defined sequence of amino acids and the biological properties resulting therefrom. Thus, a gene encodes a protein if transcription and translation of mRNA corresponding to that gene produces the protein in a cell or other biological system. Both the coding strand, the nucleotide sequence of which is identical to the mRNA sequence and is usually provided in sequence listings, and the non-coding strand, used as the template for transcription of a gene or cDNA, can be referred to as encoding the protein or other product of that gene or cDNA.

[0065] As used herein, the term “fragment,” as applied to a nucleic acid, refers to a subsequence of a larger nucleic acid. A “fragment” of a nucleic acid can be at least about 15 nucleotides in length; for example, at least about 50 nucleotides to about 100 nucleotides; at least about 100 to about 500 nucleotides, at least about 500 to about 1000 nucleotides; at least about 1000 nucleotides to about 1500 nucleotides; about 1500 nucleotides to about 2500 nucleotides; or about 2500 nucleotides (and any integer value in between). As used herein, the term “fragment,” as applied to a protein, polypeptide or peptide, refers to a subsequence of a larger protein or peptide. A “fragment” of a protein, polypeptide, or peptide can be at least about 5 amino acids in length; for example, at least about 10 amino acids in length; at least about 20 amino acids in length; at least about 50 amino acids in length; at least about 100 amino acids in length; at least about 200 amino acids in length; or at least about 300 amino acids in length (and any integer value in between).

[0066] The term “gene” refers to a nucleic acid (e.g., DNA) sequence that includes coding sequences necessary for the production of a polypeptide, precursor, or RNA (e.g., mRNA). The polypeptide may be encoded by a full-length coding sequence or by any portion of the coding sequence so long as the desired activity or functional property (e.g., enzymatic activity, ligand binding, signal transduction, immunogenicity, etc.) of the full-length or fragment is retained. The term also encompasses the coding region of a structural gene and the sequences located adjacent to the coding region on both the 5' and 3' ends for a distance of about 2 kb or more on either end such that the gene corresponds to the length of the full-length mRNA and 5' regulatory sequences which influence the transcriptional properties of the gene. Sequences located 5' of the coding region and present on the mRNA are referred to as 5'-untranslated sequences. The 5'-untranslated sequences usually contain the regulatory sequences. Sequences located 3' or downstream of the coding region and present on the mRNA are referred to as 3'- untranslated sequences. The term “gene” encompasses both cDNA and genomic forms of a gene. A genomic form or clone of a gene contains the coding region interrupted with non-coding sequences termed “introns” or “intervening regions” or “intervening sequences.” Introns are segments of a gene that are transcribed into nuclear RNA (hnRNA); introns may contain regulatory elements such as enhancers. Introns are removed or “spliced out” from the nuclear or primary transcript; introns therefore are absent in the messenger RNA (mRNA) transcript. The mRNA functions during translation to specify the sequence or order of amino acids in a nascent polypeptide.

[0067] “Homologous” refers to the sequence similarity or sequence identity between two polypeptides or between two nucleic acid molecules. When a position in both of the two compared sequences is occupied by the same base or amino acid monomer subunit, e g., if a position in each of two DNA molecules is occupied by adenine, then the molecules are homologous at that position. The percent of homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the number of positions compared X 100. For example, if 6 of 10 of the positions in two sequences are matched or homologous then the two sequences are 60% homologous. By way of example, the DNA sequences ATTGCC and TATGGC share 50% homology. Generally, a comparison is made when two sequences are aligned to give maximum homology.

[0068] “Isolated” means altered or removed from the natural state. For example, a nucleic acid or a polypeptide naturally present in a living animal is not “isolated,” but the same nucleic acid or polypeptide partially or completely separated from the coexisting materials of its natural state is “isolated.” An isolated nucleic acid or protein can exist in substantially purified form, or can exist in a non-native environment such as, for example, a host cell.

[0069] An “isolated nucleic acid” refers to a nucleic acid segment or fragment which has been separated from sequences which flank it in a naturally occurring state, e.g., a DNA fragment which has been removed from the sequences which are normally adjacent to the fragment, e.g., the sequences adjacent to the fragment in a genome in which it naturally occurs. The term also applies to nucleic acids which have been substantially purified from other components which naturally accompany the nucleic acid, e.g., RNA or DNA or proteins, which naturally accompany it in the cell. The term therefore includes, for example, a recombinant DNA which is incorporated into a vector, into an autonomously replicating plasmid or virus, or into the genomic DNA of a prokaryote or eukaryote, or which exists as a separate molecule (e.g., as a cDNA or a genomic or cDNA fragment produced by PCR or restriction enzyme digestion) independent of other sequences. It also includes a recombinant DNA which is part of a hybrid gene encoding additional polypeptide sequence.

[0070] The term “label” when used herein refers to a detectable compound or composition that is conjugated directly or indirectly to a probe to generate a “labeled” probe. The label may be detectable by itself (e.g., radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition that is detectable (e.g., avidin-biotin). In some instances, primers can be labeled to detect a PCR product. By the term “modulating,” as used herein, is meant mediating a detectable increase or decrease in the activity and / or level of a mRNA, polypeptide, or a response in a subject compared with the activity and / or level of a mRNA, polypeptide or a response in the subject in the absence of a treatment or compound, and / or compared with the activity and / or level of a mRNA, polypeptide, or a response in an otherwise identical but untreated subject. The term encompasses activating, inhibiting and / or otherwise affecting a native signal or response thereby mediating a beneficial therapeutic response in a subject, preferably, a human.

[0071] A “mutation,” as used herein, refers to a change in nucleic acid or polypeptide sequence relative to a reference sequence (which is preferably a naturally- occurring normal or “wild-type” sequence), and includes translocations, deletions, insertions, and substitutions / point mutations. A “mutant” as used herein, refers to either a nucleic acid or protein comprising a mutation.

[0072] A “nucleic acid” refers to a polynucleotide and includes polyribonucleotides and poly-deoxyribonucleotides. Nucleic acids according to the present invention may include any polymer or oligomer of pyrimidine and purine bases, preferably cytosine, thymine, and uracil, and adenine and guanine, respectively. (See Albert L. Lehninger, Principles of Biochemistry, at 793-800 (Worth Pub. 1982) which is herein incorporated in its entirety for all purposes). Indeed, the present invention contemplates any deoxyribonucleotide, ribonucleotide or peptide nucleic acid component, and any chemical variants thereof, such as methylated, hydroxymethylated or glucosylated forms of these bases, and the like. The polymers or oligomers may be heterogeneous or homogeneous in composition, and may be isolated from naturally occurring sources or may be artificially or synthetically produced. In addition, the nucleic acids may be DNA or RNA, or a mixture thereof, and may exist permanently or transitionally in single-stranded or double-stranded form, including homoduplex, heteroduplex, and hybrid states.

[0073] An “oligonucleotide” or “polynucleotide” is a nucleic acid ranging from at least 2, preferably at least 8, 15 or 25 nucleotides in length, but may be up to 50, 100, 1000, or 5000 nucleotides long or a compound that specifically hybridizes to a polynucleotide. Polynucleotides include sequences of deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) or mimetics thereof which may be isolated from natural sources, recombinantly produced or artificially synthesized. A further example of a polynucleotide of the present invention may be a peptide nucleic acid (PNA). (See U.S. Pat. No. 6,156,501 which is hereby incorporated by reference in its entirety.) The invention also encompasses situations in which there is a nontraditional base pairing such as Hoogsteen base pairing which has been identified in certain tRNA molecules and postulated to exist in a triple helix. “Polynucleotide” and “oligonucleotide” are used interchangeably in this disclosure. It will be understood that when a nucleotide sequence is represented herein by a DNA sequence (e.g., A, T, G, and C), this also includes the corresponding RNA sequence (e.g., A, U, G, C) in which “U” replaces “T”.

[0074] The terms “patient,” “subject,” “individual,” and the like are used interchangeably herein, and refer to any animal, or cells thereof whether in vitro or in situ, amenable to the methods described herein. In certain non-limiting embodiments, the patient, subject or individual is a human.

[0075] As used herein, the term “polymerase chain reaction” (“PCR”) refers to the method of K. B. Mullis (U.S. Pat. Nos. 4,683,195 4,683,202, and 4,965,188, hereby incorporated by reference) for increasing the concentration of a segment of a target sequence in a mixture of genomic DNA without cloning or purification. This process for amplifying the target sequence consists of introducing a large excess of two oligonucleotide primers to the DNA mixture containing the desired target sequence, followed by a precise sequence of thermal cycling in the presence of a DNA polymerase. The two primers are complementary to their respective strands of the double stranded target sequence. To effect amplification, the mixture is denatured and the primers then annealed to their complementary sequences within the target molecule. Following annealing, the primers are extended with a polymerase so as to form a new pair of complementary strands. The steps of denaturation, primer annealing and polymerase extension can be repeated many times (i.e., denaturation, annealing and extension constitute one “cycle”; there can be numerous “cycles”) to obtain a high concentration of an amplified segment of the desired target sequence. The length of the amplified segment of the desired target sequence is determined by the relative positions of the primers with respect to each other, and therefore, this length is a controllable parameter. By virtue of the repeating aspect of the process, the method is referred to as the “polymerase chain reaction” (hereinafter “PCR”). Because the desired amplified segments of the target sequence become the predominant sequences (in terms of concentration) in the mixture, they are said to be “PCR amplified”. As used herein, the terms “PCR product,” “PCR fragment”, “amplification product” or “amplicon” refer to the resultant mixture of compounds after two or more cycles of the PCR steps of denaturation, annealing and extension are complete. These terms encompass the case where there has been amplification of one or more segments of one or more target sequences.

[0076] As used herein, the term “probe” refers to an oligonucleotide (i.e., a sequence of nucleotides), whether occurring naturally as in a purified restriction digest or produced synthetically, recombinantly or by PCR amplification, that is capable of hybridizing to another oligonucleotide of interest. A probe may be single-stranded or double-stranded. Probes are useful in the detection, identification and isolation of particular gene sequences.

[0077] As used herein, the terms “peptide,” “polypeptide,” and “protein” are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that can comprise the sequence of a protein or peptide. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. The polypeptides include natural peptides, recombinant peptides, synthetic peptides, or a combination thereof.

[0078] As used herein, “polynucleotide” includes cDNA, RNA, DNA / RNA hybrid, antisense RNA, ribozyme, genomic DNA, synthetic forms, and mixed polymers, both sense and antisense strands, and may be chemically or biochemically modified to contain non-natural or derivatized, synthetic, or semi -synthetic nucleotide bases. Also, contemplated are alterations of a wild type or synthetic gene, including but not limited to deletion, insertion, substitution of one or more nucleotides, or fusion to other polynucleotide sequences.

[0079] To “prevent” a disease or disorder as the term is used herein, means to reduce the severity or frequency of at least one sign or symptom of a disease or disorder being experienced by a subject.

[0080] “Sample” or “biological sample” as used herein means a biological material isolated from a subject. The biological sample may contain any biological material suitable for detecting a mRNA, polypeptide or other marker of a physiologic or pathologic process in a subject, and may comprise fluid, tissue, cellular and / or non- cellular material obtained from the individual.

[0081] As used herein, “substantially purified” refers to being essentially free of other components. For example, a substantially purified polypeptide is a polypeptide which has been separated from other components with which it is normally associated in its naturally occurring state.

[0082] As used herein, the terms “therapy” or “therapeutic regimen” refer to those activities taken to prevent, alleviate or alter a disorder or disease state, e.g., a course of treatment intended to reduce or eliminate at least one sign or symptom of a disease or disorder using pharmacological, surgical, dietary and / or other techniques. A therapeutic regimen may include a prescribed dosage of one or more compounds or surgery. Therapies will most often be beneficial and reduce or eliminate at least one sign or symptom of the disorder or disease state, but in some instances the effect of a therapy will have non-desirable or side-effects. The effect of therapy will also be impacted by the physiological state of the subject, e.g., age, gender, genetics, weight, other disease conditions, etc.

[0083] The term “therapeutically effective amount” refers to the amount of the subject compound that will elicit the biological or medical response of a tissue, system, or subject that is being sought by the researcher, veterinarian, medical doctor or other clinician. The term “therapeutically effective amount” includes that amount of a compound that, when administered, is sufficient to prevent development of, or alleviate to some extent, one or more of the signs or symptoms of the disorder or disease being treated. The therapeutically effective amount will vary depending on the compound, the disease and its severity and the age, weight, etc., of the subject to be treated.

[0084] To “treat” a disease or disorder as the term is used herein, means to reduce the frequency or severity of at least one sign or symptom of a disease or disorder experienced by a subject.

[0085] As used herein, the term “wild-type” refers to a gene or gene product isolated from a naturally occurring source. A wild-type gene is that which is most frequently observed in a population and is thus arbitrarily designed the “normal” or “wild-type” form of the gene. In contrast, the term “modified” or “mutant” refers to a gene or gene product that displays modifications in sequence and / or functional properties (i.e., altered characteristics) when compared to the wild-type gene or gene product. It is noted that naturally occurring mutants can be isolated; these are identified by the fact that they have altered characteristics (including altered nucleic acid sequences) when compared to the wild-type gene or gene product.

[0086] Ranges: throughout this disclosure, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

[0087] Description

[0088] The present invention is based, in part, on the unexpected findings that certain peptide fragments within the sequence of the human renalase protein exhibit renalase agonist activity and that specific modifications to the certain peptide fragments within the sequence of the human renalase protein that exhibit renalase agonist activity increases their half-life in vivo and / or in vitro.

[0089] As such, in one embodiment, the present invention provides at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro halflife. In one embodiment, the at least one modified renalase agonist peptide comprises at least one fragment of full-length human renalase protein, SEQ ID NO: 4, or a variant thereof. In one embodiment, the at least one modified renalase agonist peptide comprises at least one fragment of full-length human renalase protein (SEQ ID NO: 4), wherein the at least one fragment comprises an amino acid sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 5, 6, 21, or 26.

[0090] In one embodiment, the at least one modified renalase agonist peptide comprises at least one fragment of full-length human renalase protein, SEQ ID NO: 4, or a variant thereof. In one embodiment, the at least one modified renalase agonist peptide comprises at least one fragment of full-length human renalase protein (SEQ ID NO: 4), wherein the at least one fragment comprises an amino acid sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 5, 6, 13, 14, 15, 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36.

[0091] In one embodiment, the at least one modified renalase agonist peptide is modified to comprise at least one N-terminal glycine residue. In one embodiment, the at least one modified renalase agonist peptide is amidated at the C-terminus. In one embodiment, the at least one N-terminal glycine residue is linked to at least one molecule that enhances association of the at least one modified renalase agonist peptide to human serum albumin in vivo and / or in vitro. In one embodiment, at least one molecule that enhances association of the at least one modified renalase agonist peptide to human serum albumin in vivo and / or in vitro is selected from myristoleic acid and palmitoleic acid.

[0092] In another embodiment, the at least one modified renalase agonist peptide is N-terminally linked to human serum albumin or a fragment thereof. In one embodiment, the at least one modified renalase agonist peptide is N-terminally linked to human serum albumin or a fragment thereof, wherein the human serum albumin comprises a sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 17. In one embodiment, the at least one modified renalase agonist peptide is N- terminally linked to human serum albumin or a fragment thereof through a linker wherein the linker comprises (G4S)2 (SEQ ID NO: 2). In one embodiment, the at least one modified renalase agonist peptide is N-terminally linked to human serum albumin or a fragment thereof through a linker wherein the linker comprises a sequence that is at least 90% identical to SEQ ID NO: 2. In one embodiment, the at least one modified renalase agonist peptide is C-terminally linked to LEAS. In one embodiment, the at least one modified renalase agonist peptide is C-terminally linked to a sequence that is at least 90% identical to the sequence as set forth in SEQ ID NO: 8. In one embodiment, the at least one modified renalase agonist peptide is C-terminally linked to a Strep Tag (SEQ ID NO: 3). In one embodiment, the at least one modified renalase agonist peptide is C-terminally linked to a sequence that is at least 90% identical to the amino acid sequence selected from SEQ ID NO: 3.

[0093] In one embodiment, the present invention provides at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life, wherein the increased in vivo and / or in vitro half-life comprises an in vivo and / or in vitro half-life of at least 1 day.

[0094] In one embodiment, the present invention provides a composition comprising at least one modified renalase agonist peptide, described herein, which displays increased in vivo and / or in vitro half-life wherein the composition is a pharmaceutical composition.

[0095] In one embodiment, the present invention provides a method of preventing or treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the at least one modified renalase peptide, described herein, which displays increased in vivo and / or in vitro half-life. In another embodiment, the present invention provides a method of preventing or treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising at least one modified renalase agonist peptide, described herein, which displays increased in vivo and / or in vitro half-life, and wherein the composition is a pharmaceutical composition. In one embodiment, the present invention provides a method of preventing or treating a subject having, suspected of having, or at risk for having a disease or disorder. Exemplary diseases or disorders that are treatable or preventable by way of the compositions and methods of the present invention include, but are not limited to, ischemic and toxic organ injury. In various embodiments, the compositions and methods described herein can be used to prevent or treat any disease or disorder that is preventable or treatable by renalase agonist activity.

[0096] Modified renalase agonist peptide sequences

[0097] In one embodiment the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence that is about at least 90% identical to a sequence set forth in SEQ ID NO: 5. In one embodiment the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence that is about at least 90% identical to a sequence set forth in SEQ ID NO: 6. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises the sequence as set forth in SEQ ID NO: 5. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises the sequence as set forth in SEQ ID NO: 6. In one embodiment the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence that is about at least 90% identical to a sequence set forth in SEQ ID NO: 21. In one embodiment the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence that is about at least 90% identical to a sequence set forth in SEQ ID NO: 26. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises the sequence as set forth in SEQ ID NO: 21. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises the sequence as set forth in SEQ ID NO: 26.

[0098] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence of 36 amino acids wherein the sequence is at least about 90% identical to the corresponding sequence as set forth in SEQ ID NO: 4. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence of 36 amino acid residues wherein the sequence is identical to the corresponding sequence as set forth in SEQ ID NO: 4.

[0099] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence of about 34 to about 38 amino acids wherein the sequence is at least about 90% identical to the corresponding sequence as set forth in SEQ ID NO: 4. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence of about 32 to about 40 amino acids wherein the sequence is at least about 90% identical to the corresponding sequence as set forth in SEQ ID NO: 4. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence of about 30 to about 42 amino acids wherein the sequence is at least about 90% identical to the corresponding sequence as set forth in SEQ ID NO: 4. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence of about 28 to about 42 amino acids wherein the sequence is at least about 90% identical to the corresponding sequence as set forth in SEQ ID NO: 4. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence of about 26 to about 44 amino acids wherein the sequence is at least about 90% identical to the corresponding sequence as set forth in SEQ ID NO: 4. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence of about 24 to about 46 amino acids wherein the sequence is at least about 90% identical to the corresponding sequence as set forth in SEQ ID NO: 4. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence of about 22 to about 48 amino acids wherein the sequence is at least about 90% identical to the corresponding sequence as set forth in SEQ ID NO: 4. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence of about 20 to about 50 amino acids wherein the sequence is at least about 90% identical to the corresponding sequence as set forth in SEQ ID NO: 4. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence of about 12 to about 20 amino acid residues wherein the sequence is at least about 90% identical to the corresponding sequence as set forth in SEQ ID NO: 4. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence of about 4 to about 12 amino acid residues wherein the sequence is at least about 90% identical to the corresponding sequence as set forth in SEQ ID NO: 4.

[0100] In one embodiment, the present invention provides a nucleic acid molecule encoding one or more modified renalase agonist peptides described herein. In various embodiments, the nucleic acid molecule is an isolated nucleic acid molecule. For example, in certain embodiments, the nucleic acid molecule is a DNA, RNA, or mRNA, encoding one or more of the modified renalase agonist peptides described herein.

[0101] In one embodiment, the nucleic acid molecule encodes one more of the modified renalase agonist peptides that comprise at least one amino acid sequence selected from SEQ ID NOs: 5, 6, 21, and 26.

[0102] In one embodiment, the nucleic acid molecule comprises a nucleic acid sequence encoding an amino acid sequence having substantial homology to at least one amino acid sequence selected from SEQ ID NOs: 5, 6, 21, and 26, or a variant thereof. For example, in certain one embodiments, the nucleic acid molecule comprises a nucleic acid sequence encoding at least one amino acid sequence that is at least about 60%, 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% identical to the amino acid sequence selected from SEQ ID NOs: 5, 6, 21, and 26. In some embodiments, the nucleic acid molecule comprises a nucleic acid sequence encoding at least one amino acid sequence as set forth in SEQ ID NOs: 5, 6, 21, and 26.

[0103] In one embodiment the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence that is about at least 90% identical to a sequence set forth in SEQ ID NOs: 5, 6, 13, 14, 15, 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, and 36. In one embodiment, the nucleic acid molecule comprises a nucleic acid sequence encoding an amino acid sequence having substantial homology to at least one amino acid sequence selected from SEQ ID NOs: 5, 6, 13, 14, 15, 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, and 36, or a variant thereof. For example, in certain one embodiments, the nucleic acid molecule comprises a nucleic acid sequence encoding at least one amino acid sequence that is at least about 60%, 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% identical to the amino acid sequence selected from SEQ ID NOs: 5, 6, 13, 14, 15, 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, and 36. In some embodiments, the nucleic acid molecule comprises a nucleic acid sequence encoding at least one amino acid sequence as set forth in SEQ ID NOs: 5, 6, 13, 14, 15, 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, and 36.

[0104] Modifications of renalase peptides

[0105] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises at least one modification. In one embodiment, the at least one modification comprises at least one N- terminal glycine residue. In one embodiment the at least one modification comprises C- terminal amidation. In one embodiment, the at least one N-terminal glycine residue is linked to at least one molecule that enhances association of the at least one modified renalase agonist peptide with human serum albumin in vivo and / or in vitro. In one embodiment, the at least one molecule that enhances association of the at least one modified renalase agonist peptide with human serum albumin in vivo and / or in vitro is selected from myristoleic acid and palmitoleic acid.

[0106] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises at least one modification. In one embodiment the at least one modification comprises the addition of at least one glycine residue at any position of at least one amino acid sequence as set forth in SEQ ID NOs: 5, 6, 21, and 26. In one embodiment, the addition of at least one glycine residue at any position of at least one amino acid sequence as set forth in SEQ ID NOs: 5, 6, 21, and 26 comprises the addition of at least one glycine residue at any position of at least one amino acid sequence as set forth in SEQ ID NOs: 5, 6, 21, and 26, wherein a glycine residue is non-naturally occurring. In one embodiment, the addition of at least one glycine residue at any position of at least one amino acid sequence as set forth in SEQ ID NOs: 5, 6, 21, and 26, comprises at least one added glycine residue. In one embodiment, the at least one added glycine residue is linked to at least one molecule that enhances association of the at least one modified renalase agonist peptide with human serum albumin in vivo and / or in vitro. In one embodiment, the at least one molecule that enhances association of the at least one modified renalase agonist peptide with human serum albumin in vivo and / or in vitro is selected from myristoleic acid and palmitoleic acid.

[0107] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises at least one modification. In one embodiment, the at least one modification comprises the addition of at least one amino acid at any position of at least one amino acid sequence as set forth in SEQ ID NOs: 5, 6, 21, and 26. In one embodiment, the addition of at least one amino acid at any position of at least one amino acid sequence as set forth in SEQ ID NOs: 5, 6, 21, and 26 that enhances association of the at least one modified renalase agonist peptide with human serum albumin in vivo and / or in vitro.

[0108] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises a sequence that is at least about 90% identical to at least one sequence as set forth in SEQ ID NOs: 13-16. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises at least one sequence as set forth in SEQ ID NOs: 13-16.

[0109] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises at least one modification. In one embodiment, the at least one modification comprises at least one chemical modification on at least one amino acid within the at least one modified renalase agonist peptide which increases the stability of the at least one modified renalase agonist peptide in vivo and / or in vitro. In one embodiment, the at least one chemical modification comprises polyubiquitination, ubiquitination, phosphorylation, glycosylation, sumoylation, acetylation, S-nitrosylation or nitrosylation, citrullination or deimination, neddylation, the addition of OClcNAc, ADP-ribosylation, methylation, hydroxymethylation, fattenylation, UFMylation, prenylation, myristoylation, S- palmitoylation, tyrosine sulfation, formylation, carboxylation, isoprenylation, farnesylation, geranylgeranylation, glypiation, lipoylation, the addition of a flavin moiety, the addition of heme C, phosphopantetheinylation, acylation (e.g., O-acylation (esters), N-acylation (amides), S-acylation (thioesters)), amidation, the addition of one or more amino acids (e.g., addition of N-terminal glycine), arginylation, polyglutamylation, polyglycylation, butyrylation, gamma-carboxylation, addition of N-acetylglucosamine to serine or threonine residues in a -glycosidic linkage, polysialylation, malonylation, hydroxylation, iodination, nucleotide addition (e g., ADP-ribosylation), adenylylation, phosphate ester (O-linked) or phosphoramidate (N-linked) formation, uridylylation, propionylation, pyroglutamate formation, S-glutathionylation, S-nitrosylation, S- sulfenylation, S-sulfinylation, S-sulfonylation, succinylation, glycation, carb amyl ati on, carbonylation, biotinylation, oxidation, pegylation, ISGylation, pupylation, citrullination, deamidation, eliminylation, disulfide bridge formation, isoaspartate formation, racemization, or any combination thereof.

[0110] In one embodiment, the at least one modification comprises any chemical modification known in the art to increase the in vivo and / or in vitro half-life of a protein, polypeptide, or peptide.

[0111] In some embodiments, the present invention provides a fusion peptide comprising a modified renalase agonist peptide, wherein the modified renalase agonist peptide is linked to an additional amino acid sequence or peptide. In some embodiments, the modified renalase agonist peptide is N-terminally linked to human serum albumin. In some embodiments, the human serum albumin is at least 90% identical to the amino acid sequence of SEQ ID NO: 17. In some embodiments, the fusion peptide comprises a linker between the modified renalase agonist peptide and the additional amino acid sequence or peptide. In some embodiments, the half-life of the fusion peptide is increased as compared to an unmodified renalase peptide. In some embodiments, the modified renalase agonist peptide of the fusion peptide comprises SEQ ID NOs: 5, 6, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, or 36. In some embodiments, the modified renalase agonist peptide of the fusion peptide comprises SEQ ID NOs: 5, 6, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36, or any combination thereof.

[0112] Modification of renalase peptide via linking to large molecule

[0113] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises at least one modification. In one embodiment, the at least one modification comprises linking the at least one modified renalase agonist peptide to at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide.

[0114] In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises human serum albumin (SEQ ID NO: 17), a variant thereof or a fragment thereof. In one embodiment, the human serum albumin (SEQ ID NO: 17), a variant thereof or a fragment thereof is linked to the N-terminus of the at least one modified renalase agonist peptide. In one embodiment, the human serum albumin (SEQ ID NO: 17), a variant thereof or a fragment thereof is linked to the C-terminus of the at least one modified renalase agonist peptide. In one embodiment, the human serum albumin (SEQ ID NO: 17), a variant thereof or a fragment thereof is linked to both the N-terminus and the C-terminus of the at least one modified renalase agonist peptide.

[0115] In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a variant of human serum albumin, wherein the variant comprises an amino acid sequence that is at least about 70% identical to the sequence set forth in SEQ ID NO: 17. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a variant of human serum albumin, wherein the variant comprises an amino acid sequence that is at least about 75% identical to the sequence set forth in SEQ ID NO: 17. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a variant of human serum albumin, wherein the variant comprises an amino acid sequence that is at least about 80% identical to the sequence set forth in SEQ ID NO: 17. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a variant of human serum albumin, wherein the variant comprises an amino acid sequence that is at least about 85% identical to the sequence set forth in SEQ ID NO: 17. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a variant of human serum albumin, wherein the variant comprises an amino acid sequence that is at least about 90% identical to the sequence set forth in SEQ ID NO: 17.

[0116] In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a fragment of human serum albumin, wherein the fragment comprises about 150 to about 250 amino acid residues of the sequence as set forth in SEQ ID NO: 17. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a fragment of human serum albumin, wherein the fragment comprises about 250 to about 400 amino acid residues of the sequence as set forth in SEQ ID NO: 17. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a fragment of human serum albumin, wherein the fragment comprises about 400 to about 609 amino acid residues of the sequence as set forth in SEQ ID NO: 17.

[0117] In one embodiment, the present invention provides a nucleic acid molecule encoding the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide. In various embodiments, the nucleic acid molecule is an isolated nucleic acid molecule. For example, in certain embodiments, the nucleic acid molecule is a DNA, RNA, or mRNA, encoding one or more of the modified renalase agonist peptides described herein.

[0118] In one embodiment, the nucleic acid molecule encodes the amino acid sequence of the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprising the sequence as set forth in SEQ ID NO: 17.

[0119] In one embodiment, the nucleic acid molecule comprises a nucleic acid sequence encoding an amino acid sequence having substantial identity to the amino acid sequence as set forth in SEQ ID NO: 17, or a variant thereof. For example, in certain one embodiments, the nucleic acid molecule comprises a nucleic acid sequence encoding at least one amino acid sequence that is at least about 60%, 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% identical to the amino acid sequence as set forth in SEQ ID NO: 17. In some embodiments, the nucleic acid molecule comprises a nucleic acid sequence encoding the amino acid sequence as set forth in SEQ ID NO: 17.

[0120] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises at least one modification. In one embodiment, the at least one modification comprises linking the at least one modified renalase agonist peptide to at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide is linked to the N-terminal of the at least one modified renalase agonist peptide via a linker, wherein the linker comprises an amino acid sequence that is at least about 90% identical to the sequence set forth in SEQ ID NO: 2. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide is linked to the N-terminal of the at least one modified renalase agonist peptide via a linker, wherein the linker comprises an amino acid sequence as set forth in SEQ ID NO: 2. In one embodiment, the at least one modified renalase agonist peptide is linked to the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide via a linker, wherein the linker is any suitable linker known in the field.

[0121] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises at least one modification. In one embodiment, the at least one modification comprises linking the at least one modified renalase agonist peptide to a C-terminal tag. In one embodiment, the at least one modification comprises linking the at least one modified renalase agonist peptide to a C-terminal tag, wherein the C-terminal tag is a Strep tag, wherein the C- terminal Step tag comprises the sequence as set forth in SEQ ID NO: 3, and wherein the C-terminal Strep tag is linked to the at least one modified renalase agonist peptide via a linker. In one embodiment, the at least one modification comprises linking the at least one modified renalase agonist peptide to a C-terminal tag, wherein the C-terminal tag is any suitable tag known in the field.

[0122] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises an N-terminally linked molecule, linked to the at least one renalase agonist peptide via a linker, and a C- terminal Strep tag, linked to the at least one renalase agonist peptide via a linker, wherein the N-terminally linked molecule comprises the sequence set forth in SEQ ID NO: 17, or a variant or fragment thereof, wherein the linker comprises the sequence set forth in SEQ ID NO: 2, and wherein the C-terminal Step tag comprises the sequence set forth in SEQ ID NO: 3.

[0123] In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises mouse serum albumin (SEQ ID NO: 1), a variant thereof or a fragment thereof. In one embodiment, the mouse serum albumin (SEQ ID NO: 1), a variant thereof or a fragment thereof is linked to the N-terminus of the at least one modified renalase agonist peptide. In one embodiment, the mouse serum albumin (SEQ ID NO: 1), a variant thereof or a fragment thereof is linked to the C-terminus of the at least one modified renalase agonist peptide. In one embodiment, the mouse serum albumin (SEQ ID NO: 1), a variant thereof or a fragment thereof is linked to both the N-terminus and the C-terminus of the at least one modified renalase agonist peptide.

[0124] In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a variant of mouse serum albumin, wherein the variant comprises an amino acid sequence that is at least about 70% identical to the sequence set forth in SEQ ID NO: 1. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a variant of mouse serum albumin, wherein the variant comprises an amino acid sequence that is at least about 75% identical to the sequence set forth in SEQ ID NO: 1. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a variant of mouse serum albumin, wherein the variant comprises an amino acid sequence that is at least about 80% identical to the sequence set forth in SEQ ID NO: 1. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a variant of mouse serum albumin, wherein the variant comprises an amino acid sequence that is at least about 85% identical to the sequence set forth in SEQ ID NO: 1. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a variant of mouse serum albumin, wherein the variant comprises an amino acid sequence that is at least about 90% identical to the sequence set forth in SEQ ID NO: 1.

[0125] In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a fragment of mouse serum albumin, wherein the fragment comprises about 150 to about 250 amino acid residues of the sequence as set forth in SEQ ID NO: 1. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a fragment of mouse serum albumin, wherein the fragment comprises about 250 to about 400 amino acid residues of the sequence as set forth in SEQ ID NO: 1. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises a fragment of mouse serum albumin, wherein the fragment comprises about 400 to about 609 amino acid residues of the sequence as set forth in SEQ ID NO: 1.

[0126] In one embodiment, the present invention provides a nucleic acid molecule encoding the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide. In various embodiments, the nucleic acid molecule is an isolated nucleic acid molecule. For example, in certain embodiments, the nucleic acid molecule is a DNA, RNA, or mRNA, encoding one or more of the modified renalase agonist peptides described herein.

[0127] In one embodiment, the nucleic acid molecule encodes the amino acid sequence of the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprising the sequence as set forth in SEQ ID NO: 1.

[0128] In one embodiment, the nucleic acid molecule comprises a nucleic acid sequence encoding an amino acid sequence having substantial identity to the amino acid sequence as set forth in SEQ ID NO: 1, or a variant thereof. For example, in certain one embodiments, the nucleic acid molecule comprises a nucleic acid sequence encoding at least one amino acid sequence that is at least about 60%, 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% identical to the amino acid sequence as set forth in SEQ ID NO: 1. In some embodiments, the nucleic acid molecule comprises a nucleic acid sequence encoding the amino acid sequence as set forth in SEQ ID NO: 1.

[0129] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises at least one modification. In one embodiment, the at least one modification comprises linking the at least one modified renalase agonist peptide to at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide is linked to the N-terminal of the at least one modified renalase agonist peptide via a linker, wherein the linker comprises an amino acid sequence that is at least about 90% identical to the sequence set forth in SEQ ID NO: 2. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide is linked to the N-terminal of the at least one modified renalase agonist peptide via a linker, wherein the linker comprises an amino acid sequence as set forth in SEQ ID NO: 2. In one embodiment, the at least one modified renalase agonist peptide is linked to the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide via a linker, wherein the linker is any suitable linker known in the field.

[0130] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises at least one modification. In one embodiment, the at least one modification comprises linking the at least one modified renalase agonist peptide to a C-terminal tag. In one embodiment, the at least one modification comprises linking the at least one modified renalase agonist peptide to a C-terminal tag, wherein the C-terminal tag is a Strep tag, wherein the C- terminal Step tag comprises the sequence as set forth in SEQ ID NO: 3, and wherein the C-terminal Strep tag is linked to the at least one modified renalase agonist peptide via a linker. In one embodiment, the at least one modification comprises linking the at least one modified renalase agonist peptide to a C-terminal tag, wherein the C-terminal tag is any suitable tag known in the field.

[0131] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises an N-terminally linked molecule, linked to the at least one renalase agonist peptide via a linker, and a C- terminal Strep tag, linked to the at least one renalase agonist peptide via a linker, wherein the N-terminally linked molecule comprises the sequence set forth in SEQ ID NO: 1, or a variant or fragment thereof, wherein the linker comprises the sequence set forth in SEQ ID NO: 2, and wherein the C-terminal Step tag comprises the sequence set forth in SEQ ID NO: 3.

[0132] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises an N-terminally linked molecule, wherein the N-terminally linked molecule is N-terminally linked to a signal peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises an N-terminally linked molecule, wherein the N-terminally linked molecule is N-terminally linked to a signal peptide, and wherein the signal peptide is any known signal peptide in the field.

[0133] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises at least one modification. In one embodiment, the at least one modification comprises linking the at least one modified renalase agonist peptide to at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide. In one embodiment, the at least one molecule is any molecule known in the field to increase the in vivo and / or in vitro half-life of a protein, polypeptide or peptide. In one embodiment, the at least one molecule which increases the in vivo and / or in vitro half-life of the at least one modified renalase agonist peptide comprises cholesterol, one or more lipid molecules, one or more fatty acids, one or more antibody Fc domains, one or more unstructured, hydrophilic amino acid polymers that are functional analogs of poly(ethylene)glycol (PEG) (e.g., XTEN and PAS), CovX-Body™ (e.g., CVX-096, CVX-060, CVX-045), or any combination thereof.

[0134] Additionally, poly(ethylene)glycol (PEG) or bis-poly(ethylene)glycol (Bis-PEG) of various average molecular weights (e.g., between 500 and 20,000 amu) may be attached to either terminus of the peptide to promote longer-term activity of the peptide, as is well-known in the pharmaceutical art. Poly(ethylene)glycol is an amphiphilic polymer consisting of repeating units of ethylene oxide which may be assembled in linear or branched structures to give a range of PEGs with different configurations and molecular weights. PEG must be activated in order to be covalently conjugated to appropriate sites in biopharmaceutical compounds (including peptides and proteins) thereby improving their pharmacologic and pharmaceutical properties. In addition to improving solubility, conjugation with PEG can protect the biopharmaceutical compound from the host’s immune system thereby reducing immunogenicity and antigenicity and prolonging biological half-life. The resulting PEGylated pharmaceuticals may be used at reduced dosage and frequency without diminished efficacy. PEG is available in a variety of average molecular weights and may be functionalized at one end (the other end is usually protected as the methoxy). Alternatively, both ends of the polymer may be functionalized resulting in homobifunctional or heterobifunctional derivatives which can be used for linking two entities. A review of both first- and second- generation PEG derivatives, with more varied and efficient functional groups for conjugation to peptides and proteins has been described by Roberts, Adv. Drug Deliv. Rev., 54, 459 (2002). Third generation PEGylating agents have been developed where the polymer has been branched and may offer additional advantages for protecting proteins from proteolysis and further reducing immunogenicity and antigenicity. Although [PEG] is commonly used for conjugation to peptides and proteins, other suitably functionalized polymers, including, but not limited to carbohydrate moieties, that have appropriate flexibility can also be used and has been reviewed by Sola and Griebenow, J. Pharm. Sci., 98, 1223 (2009) and by Witteloostuijn, Pedersen, and Jensen, ChemMedChem, 11, 2474 (2016), both of which are incorporated herein by reference.

[0135] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life comprises at least one modification. In one embodiment, the at least one modification comprises linking the at least one modified renalase agonist peptide to any molecule known in the art to increase the in vivo and / or in vitro half-life of a protein, polypeptide, or peptide.

[0136] Additional modified renalase agonist peptides

[0137] In some embodiments, the stability of the at least one modified renalase agonist peptide can be increased by replacing the cysteine residue at position 220 of SEQ ID NO: 4 by a different appropriately selected amino acid. In addition, the modified peptide fragments have increased potency as compared to unmodified fragment

[0138] The amino acid at position 220 may be selected from the residue of an amino acid selected from: wherein R1 and R2 are independently H; Cl to C8 n-alkyl optionally substituted by hydroxyl; C3 to C8 branched alkyl optionally substituted by hydroxyl; C4 to C8 doubly branched alkyl optionally substituted by hydroxyl; C3 to C5 cycloalkyl optionally substituted by hydroxyl or methyl or both at any position or positions and which includes all structurally feasible stereoisomeric entities; CH2-C3 to C5 cycloalkyl optionally substituted by hydroxyl or methyl or both at any position or positions and R1 and R2 may be linked together as (CH2)n optionally substituted at any position by methyl or hydroxyl or both, wherein n is 2, 3, 4, or 5; and X220l- "'"OH wherein Y is (CH2)n optionally substituted at any position or positions by methyl or hydroxyl or both, provided that the carbon bearing the amino group may be substituted only by methyl, wherein n is 2, 3, 4, 5; cis- or trans- 1,2-cy cl opropaned iyl, cis- or trans- 1,2-cyclobutanediyl, cis- or trans-l,3-cyclobutanediyl, cis- or trans-l,2-cyclopentanediyl, cis- or trans- 1,3 -cyclopentanediyl, cis- or trans- 1,2-cy cl ohexanediyl, cis- or trans-1,3- cyclohexanediyl, or cis- or trans- 1,4-cyclohexanediyl optionally substituted by at any position or positions by methyl or hydroxyl or both provided that the carbon bearing the amino group may be substituted only by methyl and which includes all structurally feasible diastereoisomeric entities.

[0139] The at least one modified renalase agonist peptides of the invention comprise at least one fragment of the sequence as set forth in SEQ ID NO: 4 including positions 220-229 with X220as described above and comprising additional amino acids at the appropriate positions of SEQ ID NO: 4 on one or either side of positions 220-229, the peptides generally varying in length from about 49 amino acids to about 20 amino acid residues, although they may be longer as desired.

[0140] Selected amino acids for substitution at position 220 of the peptides of the invention include glycine, serine, alanine, leucine, valine, isoleucine, norleucine, betaalanine, cyclopropyl-glycine, (cyclopropylmethyl)- glycine, and other hydrophobic amino acids as well as the D-amino acid enantiomers of the described amino acids.

[0141] Additionally, the Lys229may be replaced with its D-isomer to afford additional stability. Other amino acids, including Lys205, Arg222, Lys230and / or Arg231may also be replaced with the corresponding D-amino acids for similar enhancement of stability to enzymatic degradation.

[0142] In one embodiment, there are provided peptides comprising R-X220- Ile221- Arg222-Phe223-Val234-Ser225-Ile226-Asp227-Asn228-Lys229-R' wherein the superscripts represent positions within SEQ ID NO: 4, R is selected from Ac-Ala-Gly-Thr-, Ac-Gly- Thr-, Ac-Thr-, Ac- , Ac-Z, H-, H-Z, B-Z- and B, wherein Z is selected from one or more of the amino acid residues at positions 205-219 of SEQ ID NO: 4, R' is selected from - NH2, Z'-NH2, -B, .and Z'-B, wherein Z' is selected from one or more of the amino acid residues at positions 230-253 of SEQ ID NO: 4, B is PEG or bis-PEG, and X is the residue of an amino acid selected from: wherein R1 and R2 are independently H; Cl to Can-alkyl optionally substituted by hydroxyl; C3 to C8 branched alkyl optionally substituted by hydroxyl; C4 to C8 doubly branched alkyl optionally substituted by hydroxyl; C3 to C5 cycloalkyl optionally substituted by hydroxyl or methyl or both at any position or positions; CH2-C3 to C5 cycloalkyl optionally substituted by hydroxyl or methyl or both at any position or positions and R1 and R2 may be linked together as (CH2)n optionally substituted at any position by methyl or hydroxyl or both, wherein n is 2, 3, 4, or 5; and wherein Y is (CH2)n optionally substituted at any position or positions by methyl or hydroxyl or both, provided that the carbon bearing the amino group may be substituted only by methyl, wherein n is 2, 3, 4, 5; 1 ,2-cyclopropanediyl, 1 ,2- cyclobutanediyl, 1 ,3- cyclobutanediyl, 1 ,2-cyclopentanediyl, 1 ,3- cyclopentanediyl, 1 ,2-cyclohexanediyl, 1 ,3-cyclohexanediyl, or 1 ,4- cyclohexanediyl optionally substituted by at any position or positions by methyl or hydroxyl or both, provided that the carbon bearing the amino group may be substituted only by methyl and provided that the peptide comprises at least 20 amino acid residues.

[0143] In another embodiment are provided peptides having the sequence R- Lys205-IIe206-Asp207-Val208-Pro209-Trp210-Ala211-Gly212-Gln213-Tyr214- Ile215-Thr216-Ser217-Asn218-Pro219-x220-IIe221-Arg222-Phe223-Val224-Ser225- IIe226-Asp227- Asn228-Lys229-Lys230-Arg231-Asn232-Ile233-Glu234-Ser235-Ser236-Glu237-Ile238-Gly239-Pro240- Ser241-Leu242-Val243-Ile244-His245-Thr246-Thr247-Val248-Pro249-Phe250-Gly251-Val252-Thr253- R' wherein R is selected from Ac-Ala-Gly-Thr-, AcGly-Thr-, Ac-Thr-, Ac-, H-, and PEG, R1is selected from -Tyr-Leu-Glu-NH2, Tyr-Leu-NH2, -Tyr-NH2, -NH2, -OH, and PEG, and X220 is selected from Gly, Ser, Ala, Leu, Vai, lie, Nie, 0-Ala cyclopropyl -Gyl, (cyclopropylmethyl)-Gly, and Aib.

[0144] Thus, peptides of the invention include: [X220] -Ac-Renal ase A (205- 240)- NH2 (SEQ ID NO: 18), [X220]-Ac-Renalase A (214-253)-NH2 , (SEQ ID NO: 19) [X220]- Ac-Renalase A (214-240)-NH2 (SEQ ID NO: 20), and [X220]-Ac- Renalase A (205-253)- NH2 (SEQ ID NO: 21), wherein X is selected from glycine, serine, alanine, leucine, valine, isoleucine, norleucine, cyclopropyl-glycine, (cyclopropylmethyl)-glycine, and beta-alanine.

[0145] Representative compounds of the present invention include but are not limited to: [Al a220] -Ac-Renal ase A (205-240)-NH2 (SEQ ID NO: 22), [Ala220]-Ac- Renalase A (214-253)-NH2 (SEQ ID NO: 23), [Ala220]-Ac-Renalase A (214-240)-NH2 (SEQ ID NO: 24), [Ala220]-Ac-Renalase A (205-253)-NH2 (SEQ ID NO: 25), [Val220]- Ac-Renalase A (214-240)-NH2 (SEQ ID NO: 26), [Ser220]-Ac-Renalase A (214-253)- NH2 (SEQ ID NO: 27), [Ala220, D-Lys229]-Ac-Renalase A (205-240)-NH2 (SEQ ID NO: 28), [Ser220]-Ac-Renalase A (205-240)-NH2 (SEQ ID NO: 29), [Ala220]-Ac-Renalase A (214-234)-NH2 (SEQ ID NO: 30), [Ala220]-Ac-Renalase A (220-239)-NH2 (SEQ ID NO: 31), [Gly220]-Ac-Renalase A (205-240)-NH2 (SEQ ID NO: 32), [Gly220]-Ac-Renalase A (214-253)-NH2 (SEQ ID NO: 33), [cyclopropyl-Gly220]-Ac-Renalase A (205-253)-NH2 (SEQ ID NO: 34), [(cyclopropylmethyl)-Gly220]-Ac-Renalase A (214-240)-NH2 (SEQ ID NO: 35).

[0146] One representative example of a PEGylated peptide of the invention is [bis-PEGsooo]- Lys20:’-lle206-Asp207-Val208-Pro209-Trp210-Ala211-Gly212-Gln213-Tyr214- lle21:>-Thr216-Ser217-Asn218-Pro219-Ala220-lle221-Arg222-Phe223-Val234-Ser225-lle226-Asp227- Asn228-Lys229-Lys230-Arg231-Asn232-lle233-Glu234-Ser235-Ser236-Glu237-lle238-Gly239-Pro240- NH2 (SEQ ID NO: 36).

[0147] Peptides are generally prepared using solid phase synthesis, such as that described by Merrifield, J. Am. Chem. Soc., 85, 2149 (1963) although other equivalent chemical syntheses known to one of ordinary skill may be used, including liquid-phase synthesis or production biologically using recombinant technologies. Solid phase synthesis is commenced from the C-terminal end of the peptide by coupling an NH2- protected amino acid to a suitable resin. The starting material is prepared by attaching the COOH-terminal of Nalpha-9-fluorenylmethoxycarbonyl (Fmoc) amino acid to commercially available 4,4'-dimethoxybenzhydryl-amine, (Mbh)-handle, that is linked to a solid phase resin. The solid phase syntheses and coupling with Fmoc-amino acids (including suitably protected side chains for trifunctional amino acids) proceeded using a carbodiimide / HOBt mediated reaction in a stepwise elongation of the desired peptide chains. The final cleavage of side-chain protecting groups and the release of the C- terminal amide moiety was achieved by the treatment with trifluoroacetic acid in the presence of scavengers. Peptides were purified by preparative high performance liquid chromatography (>95% purity), and characterized by amino acid analysis and mass spectroscopy. Specific details regarding the synthesis of the present peptides are provided below.

[0148] Renalase agonist peptide half-life extension

[0149] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life of at least 1 day. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life of at least 1.5 days. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro halflife displays an in vivo and / or in vitro half-life of at least 2 days. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life of at least 3 days.

[0150] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 1 hour longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 2 hours longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 3 hours longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 4 hours longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 5 hours longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 6 hours longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro halflife that is at least 7 hours longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 8 hours longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 9 hours longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 10 hours longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 11 hours longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 12 hours longer than an unmodified renalase agonist peptide.

[0151] In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 1 day longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 2 days longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 3 days longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro halflife displays an in vivo and / or in vitro half-life that is at least 4 days longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 5 days longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro half-life that is at least 6 days longer than an unmodified renalase agonist peptide. In one embodiment, the at least one modified renalase agonist peptide which displays increased in vivo and / or in vitro half-life displays an in vivo and / or in vitro halflife that is at least 7 days longer than an unmodified renalase agonist peptide.

[0152] Methods of treatment and prevention

[0153] In various embodiments, the present invention includes at least one modified renalase agonist peptide, compositions thereof, and methods of use thereof to treat or prevent a disease or disorder in a subject. In various embodiments, the at least one modified renalase agonist peptide, compositions thereof, and methods of use thereof initiate a physiological response in a subject that is characteristic of an increase in renalase protein levels in circulation such as, but not limited to, metabolism of catecholamines, particularly dopamine, epinephrine and norepinephrine, thereby obtaining and / or maintaining higher or normal catecholamine levels in circulation.

[0154] In various embodiments, the compositions disclosed herein may be administered to patients in need of elevated renalase levels in circulation such as, but not limited to, prevention or treatment for a renal condition, disorder or disease; a cardiovascular condition, disorder or disease; a heart condition, disorder or disease; or a central nervous system (CNS) condition, disorder or disease; pre-eclampsia; or any combination thereof. Representative cardiovascular conditions, disorders or diseases include but are not limited to hypertension, asymptomatic left ventricular dysfunction, chronic congestive heart failure (CHF), myocardial infarction (MI), cardiac rhythm disturbance, stroke, atherosclerosis, and any combination thereof. Representative types or kinds of hypertension that can be prevented, treated or ameliorated by the methods of the present invention include but are not limited to chronic hypertension, systolic hypertension, isolated systolic hypertension, diabetic hypertension, pulmonary hypertension, acute severe hypertension, and any combination thereof. Representative renal conditions, disorders or diseases that can be prevented, treated or ameliorated by the methods of the present invention include but are not limited to end-stage renal disease (ESRD), chronic renal failure, and any combination thereof. Representative CNS conditions, disorders or diseases that can be prevented, treated or ameliorated by the methods of the present invention include but are not limited to depression, anxiety, mania, schizophrenia, post-traumatic stress disorder (PTSD), and any combination thereof.

[0155] It will be understood by one skilled in the art, based upon the disclosure provided herein, that administering at least one modified renalase agonist peptide has a comparable physiological outcome to increasing the level and / or activity of renalase protein. Thus, the present invention relates to the prevention and treatment of a disease or disorder by administration of a therapeutically effective amount of at least one modified renalase agonist peptide, to a subject in need thereof, for the treatment or prevention of a disease or disorder, or its associated signs, symptoms or pathologies.

[0156] It is understood by one skilled in the art, that an increase in the level of at least one modified renalase agonist peptide results in the effects of an increase in renalase activity. The increased level or activity of renalase can be assessed using a wide variety of methods, including those disclosed herein, as well as methods well-known in the art or to be developed in the future. That is, the routineer would appreciate, based upon the disclosure provided herein, that increasing the level or activity of renalase can be readily assessed using methods that quantifiable assess the effects of an increase in renalase activity.

[0157] One skilled in the art, based upon the disclosure provided herein, would understand that the invention is useful in subjects who, in whole (e.g., systemically) or in part (e g., locally, tissue, organ), are being or will be, treated for a disease or disorder such as ischemic or toxic organ injury. The skilled artisan will appreciate, based upon the teachings provided herein, that the diseases and disorders treatable by the compositions and methods described herein encompass any disease or disorder where in an increase in renalase activity will promote a positive therapeutic outcome.

[0158] Further, one of skill in the art would, when equipped with this disclosure and the methods exemplified herein, appreciate that the at least one modified renalase agonist peptide and compositions thereof can be administered together with at least one renalase activator to a subject in need thereof. The at least one renalase activator includes those which are known in the art, or renalase activators which are discovered in the future, as can be identified by well-known criteria in the art of pharmacology, such as the physiological results of activation of renalase as described in detail herein and / or as known in the art. Therefore, the present invention is not limited in any way to any particular renalase activator; rather, the invention encompasses those activators that would be understood by the routineer to be useful as are known in the art and as are discovered in the future. Further methods of identifying and producing a renalase activator are well known to those of ordinary skill in the art, including, but not limited, obtaining an activator from a naturally occurring source (e.g., Streptomyces sp., Pseudomonas sp., Stylotella aurantium, etc.). Alternatively, a renalase activator can be synthesized chemically. Further, the routineer would appreciate, based upon the teachings provided herein, that a renalase activator can be obtained from a recombinant organism. Compositions and methods for chemically synthesizing renalase activators and for obtaining them from natural sources are well known in the art and are described in the art. One of skill in the art will appreciate that an activator can be administered as a small molecule chemical, a protein, a nucleic acid construct encoding a protein, or combinations thereof. Numerous vectors and other compositions and methods are well known for administering a protein or a nucleic acid construct encoding a protein to cells or tissues. Therefore, the invention includes a method of administering a protein or a nucleic acid encoding a protein that is an activator of renalase. (Sambrook et al., 2012, Molecular Cloning: A Laboratory Manual; Ausubel et al., 1997, Current Protocols in Molecular Biology). It will be appreciated by one of skill in the art, when armed with the present disclosure including the methods detailed herein, that the invention is not limited to treatment of a disease or disorder once is established. Particularly, the symptoms of the disease or disorder need not have manifested to the point of detriment to the subject; indeed, the disease or disorder need not be detected in a subject before treatment is administered. That is, significant pathology from disease or disorder does not have to occur before the present invention may provide benefit. Therefore, the present invention, as described more fully herein, includes a method for preventing diseases and disorders in a subject, in that at least one modified renalase agonist peptide, as discussed elsewhere herein, can be administered to a subject prior to the onset of the disease or disorder, thereby preventing the disease or disorder from developing.

[0159] One of skill in the art, when armed with the disclosure herein, would appreciate that the prevention of a disease or disorder in a subject encompasses administering to a subject at least one modified renalase agonist peptide a preventative measure against a disease or disorder.

[0160] The invention encompasses administration of at least one modified renalase agonist peptide to practice the methods of the invention; the skilled artisan would understand, based on the disclosure provided herein, how to formulate and administer the appropriate at least one modified renalase agonist peptide to a subject. However, the present invention is not limited to any particular method of administration or treatment regimen. This is especially true where it would be appreciated by one skilled in the art, equipped with the disclosure provided herein, including the reduction to practice using an art-recognized model of ischemia-reperfusion injury, that methods of administering at least one modified renalase agonist peptide can be determined by one of skill in the pharmacological arts.

[0161] Pharmaceutical compositions and formulations

[0162] Compositions comprising at least one modified renalase agonist peptide can be formulated and administered to a subject, as now described. The invention encompasses the preparation and use of pharmaceutical compositions comprising at least one modified renalase agonist peptide as described elsewhere herein, for the treatment or prevention of a disease or disorder, and disclosed herein as an active ingredient. Such a pharmaceutical composition may consist of the active ingredient alone, in a form suitable for administration to a subject, or the pharmaceutical composition may comprise the active ingredient and one or more pharmaceutically acceptable carriers, one or more additional ingredients, or some combination of these. The active ingredient may be present in the pharmaceutical composition in the form of a physiologically acceptable ester or salt, such as in combination with a physiologically acceptable cation or anion, as is well known in the art.

[0163] Pharmaceutical compositions suitable for use in the methods described herein can include one or more of the disclosed peptides and a pharmaceutically acceptable carrier or diluent.

[0164] The composition may be formulated for intravenous, subcutaneous, intraperitoneal, intramuscular, topical, oral, buccal, nasal, pulmonary or inhalation, ocular, vaginal, or rectal administration. In some embodiments, the peptides are formulated for intravenous, subcutaneous, intraperitoneal, intramuscular administration, or targeted tissue delivery such as in a solution, suspension, emulsion, liposome formulation, etc. The pharmaceutical composition can be formulated to be an immediate- release composition, sustained-release composition, delayed-release composition, etc., using techniques known in the art.

[0165] Pharmacologically acceptable carriers for various dosage forms are known in the art. For example, excipients, lubricants, binders, and disintegrants for solid preparations are known; solvents, solubilizing agents, suspending agents, isotonicity agents, buffers, and soothing agents for liquid preparations are known. In some embodiments, the pharmaceutical compositions include one or more additional components, such as one or more preservatives, antioxidants, stabilizing agents and the like.

[0166] Additionally, the disclosed pharmaceutical compositions can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. In some embodiment, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, monostearate salts and gelatin.

[0167] Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by sterilization microfiltration. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze- drying (lyophilization) that yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[0168] Pharmaceutical compositions of the disclosure can be administered in combination with other therapeutics that are part of the current standard of care for the kidney disease, pancreatic disease or tissue injury, particularly those that may benefit from regulation of excessive immunoinflammation. For example, fenoldopam, discovered as a selective dopamine 1 receptor agonists, has been utilized for acute kidney injury and could be used with the subject peptides in treatment.

[0169] As used herein, the term “pharmaceutically-acceptable carrier” means a chemical composition with which an appropriate renalase modulator thereof, may be combined and which, following the combination, can be used to administer the appropriate renalase modulator thereof, to a subject.

[0170] The pharmaceutical compositions useful for practicing the invention may be administered to deliver a dose of between about 0.1 ng / kg / day and 100 mg / kg / day, or more.

[0171] In various embodiments, the pharmaceutical compositions useful in the methods of the invention may be administered, by way of example, systemically, parenterally, or topically, such as, in oral formulations, inhaled formulations, including solid or aerosol, and by topical or other similar formulations. In addition to the appropriate therapeutic composition, such pharmaceutical compositions may contain pharmaceutically acceptable carriers and other ingredients known to enhance and facilitate drug administration. Other possible formulations, such as nanoparticles, liposomes, resealed erythrocytes, and immunologically based systems may also be used to administer an appropriate modulator thereof, according to the methods of the invention.

[0172] The formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing the active ingredient into association with a carrier or one or more other accessory ingredients, and then, if necessary or desirable, shaping or packaging the product into a desired single- or multi-dose unit.

[0173] Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for ethical administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and perform such modification with merely ordinary, if any, experimentation.

[0174] A pharmaceutical composition of the invention may be prepared, packaged, or sold in bulk, as a single unit dose, or as a plurality of single unit doses. As used herein, a “unit dose” is discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject or a convenient fraction of such a dosage such as, for example, one-half or one-third of such a dosage.

[0175] The relative amounts of the active ingredient, the pharmaceutically acceptable carrier, and any additional ingredients in a pharmaceutical composition of the invention will vary, depending upon the identity, size, and condition of the subject treated and further depending upon the route by which the composition is to be administered. By way of example, the composition may comprise between 0.1% and 100% (w / w) active ingredient.

[0176] In addition to the active ingredient, a pharmaceutical composition of the invention may further comprise one or more additional pharmaceutically active agents.

[0177] Controlled- or sustained-release formulations of a pharmaceutical composition of the invention may be made using conventional technology.

[0178] A formulation of a pharmaceutical composition of the invention suitable for oral administration may be prepared, packaged, or sold in the form of a discrete solid dose unit including, but not limited to, a tablet, a hard or soft capsule, a cachet, a troche, or a lozenge, each containing a predetermined amount of the active ingredient. Other formulations suitable for oral administration include, but are not limited to, a powdered or granular formulation, an aqueous or oily suspension, an aqueous or oily solution, or an emulsion.

[0179] Pharmaceutically acceptable excipients used in the manufacture of pharmaceutical compositions include, but are not limited to, inert diluents, granulating and disintegrating agents, binding agents, and lubricating agents. Known dispersing agents include, but are not limited to, potato starch and sodium starch glycollate. Known surface active agents include, but are not limited to, sodium lauryl sulphate. Known diluents include, but are not limited to, calcium carbonate, sodium carbonate, lactose, microcrystalline cellulose, calcium phosphate, calcium hydrogen phosphate, and sodium phosphate. Known granulating and disintegrating agents include, but are not limited to, corn starch and alginic acid. Known binding agents include, but are not limited to, gelatin, acacia, pre-gelatinized maize starch, polyvinylpyrrolidone, and hydroxypropyl methylcellulose. Known lubricating agents include, but are not limited to, magnesium stearate, stearic acid, silica, and talc.

[0180] Liquid formulations of a pharmaceutical composition of the invention may be prepared, packaged, and sold either in liquid form or in the form of a dry product intended for reconstitution with water or another suitable vehicle prior to use.

[0181] Liquid suspensions may be prepared using conventional methods to achieve suspension of the active ingredient in an aqueous or oily vehicle. Aqueous vehicles include, for example, water and isotonic saline. Oily vehicles include, for example, almond oil, oily esters, ethyl alcohol, vegetable oils such as arachis, olive, sesame, or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin. Liquid suspensions may further comprise one or more additional ingredients including, but not limited to, suspending agents, dispersing or wetting agents, emulsifying agents, demulcents, preservatives, buffers, salts, flavorings, coloring agents, and sweetening agents. Oily suspensions may further comprise a thickening agent.

[0182] Known suspending agents include, but are not limited to, sorbitol syrup, hydrogenated edible fats, sodium alginate, polyvinylpyrrolidone, gum tragacanth, gum acacia, and cellulose derivatives such as sodium carboxymethylcellulose, methylcellulose, and hydroxypropylmethylcellulose. Known dispersing or wetting agents include, but are not limited to, naturally-occurring phosphatides such as lecithin, condensation products of an alkylene oxide with a fatty acid, with a long chain aliphatic alcohol, with a partial ester derived from a fatty acid and a hexitol, or with a partial ester derived from a fatty acid and a hexitol anhydride (e.g. polyoxyethylene stearate, heptadecaethyleneoxycetanol, polyoxyethylene sorbitol monooleate, and polyoxyethylene sorbitan monooleate, respectively). Known emulsifying agents include, but are not limited to, lecithin and acacia. Known preservatives include, but are not limited to, methyl, ethyl, or n-propyl-para-hydroxybenzoates, ascorbic acid, and sorbic acid. Known sweetening agents include, for example, glycerol, propylene glycol, sorbitol, sucrose, and saccharin. Known thickening agents for oily suspensions include, for example, beeswax, hard paraffin, and cetyl alcohol.

[0183] Liquid solutions of the active ingredient in aqueous or oily solvents may be prepared in substantially the same manner as liquid suspensions, the primary difference being that the active ingredient is dissolved, rather than suspended in the solvent. Liquid solutions of the pharmaceutical composition of the invention may comprise each of the components described with regard to liquid suspensions, it being understood that suspending agents will not necessarily aid dissolution of the active ingredient in the solvent. Aqueous solvents include, for example, water and isotonic saline. Oily solvents include, for example, almond oil, oily esters, ethyl alcohol, vegetable oils such as arachis, olive, sesame, or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin.

[0184] Powdered and granular formulations of a pharmaceutical preparation of the invention may be prepared using known methods. Such formulations may be administered directly to a subject, used, for example, to form tablets, to fill capsules, or to prepare an aqueous or oily suspension or solution by addition of an aqueous or oily vehicle thereto. Each of these formulations may further comprise one or more of dispersing or wetting agent, a suspending agent, and a preservative. Additional excipients, such as fillers and sweetening, flavoring, or coloring agents, may also be included in these formulations.

[0185] A pharmaceutical composition of the invention may also be prepared, packaged, or sold in the form of oil-in-water emulsion or a water-in-oil emulsion. The oily phase may be a vegetable oil such as olive or arachis oil, a mineral oil such as liquid paraffin, or a combination of these. Such compositions may further comprise one or more emulsifying agents such as naturally occurring gums such as gum acacia or gum tragacanth, naturally-occurring phosphatides such as soybean or lecithin phosphatide, esters or partial esters derived from combinations of fatty acids and hexitol anhydrides such as sorbitan monooleate, and condensation products of such partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate. These emulsions may also contain additional ingredients including, for example, sweetening or flavoring agents.

[0186] Methods for impregnating or coating a material with a chemical composition are known in the art, and include, but are not limited to methods of depositing or binding a chemical composition onto a surface, methods of incorporating a chemical composition into the structure of a material during the synthesis of the material (i.e., such as with a physiologically degradable material), and methods of absorbing an aqueous or oily solution or suspension into an absorbent material, with or without subsequent drying.

[0187] As used herein, “parenteral administration” of a pharmaceutical composition includes any route of administration characterized by physical breaching of a tissue of a subject and administration of the pharmaceutical composition through the breach in the tissue. Parenteral administration thus includes, but is not limited to, administration of a pharmaceutical composition by injection of the composition, by application of the composition through a surgical incision, by application of the composition through a tissue-penetrating non-surgical wound, and the like. In particular, parenteral administration is contemplated to include, but is not limited to, cutaneous, subcutaneous, intraperitoneal, intravenous, intramuscular, intraci sternal injection, and kidney dialytic infusion techniques.

[0188] Formulations of a pharmaceutical composition suitable for parenteral administration comprise the active ingredient combined with a pharmaceutically acceptable carrier, such as sterile water or sterile isotonic saline. Such formulations may be prepared, packaged, or sold in a form suitable for bolus administration or for continuous administration. Injectable formulations may be prepared, packaged, or sold in unit dosage form, such as in ampules or in multi-dose containers containing a preservative. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradable formulations. Such formulations may further comprise one or more additional ingredients including, but not limited to, suspending, stabilizing, or dispersing agents. In one embodiment of a formulation for parenteral administration, the active ingredient is provided in dry (i.e., powder or granular) form for reconstitution with a suitable vehicle (e.g., sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition.

[0189] The pharmaceutical compositions may be prepared, packaged, or sold in the form of a sterile injectable aqueous or oily suspension or solution. This suspension or solution may be formulated according to the known art, and may comprise, in addition to the active ingredient, additional ingredients such as the dispersing agents, wetting agents, or suspending agents described herein. Such sterile injectable formulations may be prepared using a non-toxic parenterally-acceptable diluent or solvent, such as water or 1,3-butane diol, for example. Other acceptable diluents and solvents include, but are not limited to, Ringer's solution, isotonic sodium chloride solution, and fixed oils such as synthetic mono- or di-glycerides. Other parentally-administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form, in a liposomal preparation, or as a component of a biodegradable polymer systems. Compositions for sustained release or implantation may comprise pharmaceutically acceptable polymeric or hydrophobic materials such as an emulsion, an ion exchange resin, a sparingly soluble polymer, or a sparingly soluble salt.

[0190] Formulations suitable for topical administration include, but are not limited to, liquid or semi-liquid preparations such as liniments, lotions, oil-in-water or water-in-oil emulsions such as creams, ointments or pastes, and solutions or suspensions. Topically-administrable formulations may, for example, comprise from about 1% to about 10% (w / w) active ingredient, although the concentration of the active ingredient may be as high as the solubility limit of the active ingredient in the solvent Formulations for topical administration may further comprise one or more of the additional ingredients described herein.

[0191] A pharmaceutical composition of the invention may be prepared, packaged, or sold in a formulation suitable for pulmonary administration via the buccal cavity. Such a formulation may comprise dry particles which comprise the active ingredient and which have a diameter in the range from about 0.5 to about 7 nanometers, and preferably from about 1 to about 6 nanometers. Such compositions are conveniently in the form of dry powders for administration using a device comprising a dry powder reservoir to which a stream of propellant may be directed to disperse the powder or using a self-propelling solvent / powder-dispensing container such as a device comprising the active ingredient dissolved or suspended in a low-boiling propellant in a sealed container. Preferably, such powders comprise particles wherein at least 98% of the particles by weight have a diameter greater than 0.5 nanometers and at least 95% of the particles by number have a diameter less than 7 nanometers. More preferably, at least 95% of the particles by weight have a diameter greater than 1 nanometer and at least 90% of the particles by number have a diameter less than 6 nanometers. Dry powder compositions preferably include a solid fine powder diluent such as sugar and are conveniently provided in a unit dose form.

[0192] Low boiling propellants generally include liquid propellants having a boiling point of below 65°F at atmospheric pressure. Generally the propellant may constitute 50 to 99.9% (w / w) of the composition, and the active ingredient may constitute 0.1 to 20% (w / w) of the composition. The propellant may further comprise additional ingredients such as a liquid non-ionic or solid anionic surfactant or a solid diluent (preferably having a particle size of the same order as particles comprising the active ingredient).

[0193] Pharmaceutical compositions of the invention formulated for pulmonary delivery may also provide the active ingredient in the form of droplets of a solution or suspension. Such formulations may be prepared, packaged, or sold as aqueous or dilute alcoholic solutions or suspensions, optionally sterile, comprising the active ingredient, and may conveniently be administered using any nebulization or atomization device. Such formulations may further comprise one or more additional ingredients including, but not limited to, a flavoring agent such as saccharin sodium, a volatile oil, a buffering agent, a surface active agent, or a preservative such as methylhydroxybenzoate. The droplets provided by this route of administration preferably have an average diameter in the range from about 0.1 to about 200 nanometers. The formulations described herein as being useful for pulmonary delivery are also useful for intranasal delivery of a pharmaceutical composition of the invention. Another formulation suitable for intranasal administration is a coarse powder comprising the active ingredient and having an average particle from about 0.2 to 500 micrometers.

[0194] Such a formulation is administered in the manner in which snuff is taken i.e. by rapid inhalation through the nasal passage from a container of the powder held close to the nares. Formulations suitable for nasal administration may, for example, comprise from about as little as 0.1% (w / w) and as much as 100% (w / w) of the active ingredient, and may further comprise one or more of the additional ingredients described herein.

[0195] A pharmaceutical composition of the invention may be prepared, packaged, or sold in a formulation suitable for buccal administration. Such formulations may, for example, be in the form of tablets or lozenges made using conventional methods, and may, for example, contain 0.1 to 20% (w / w) active ingredient, the balance comprising an orally dissolvable or degradable composition and, optionally, one or more of the additional ingredients described herein. Alternately, formulations suitable for buccal administration may comprise a powder or an aerosolized or atomized solution or suspension comprising the active ingredient. Such powdered, aerosolized, or aerosolized formulations, when dispersed, preferably have an average particle or droplet size in the range from about 0.1 to about 200 nanometers, and may further comprise one or more of the additional ingredients described herein.

[0196] A pharmaceutical composition of the invention may be prepared, packaged, or sold in a formulation suitable for ophthalmic administration. Such formulations may, for example, be in the form of eye drops including, for example, a 0.1- 1.0% (w / w) solution or suspension of the active ingredient in an aqueous or oily liquid carrier. Such drops may further comprise buffering agents, salts, or one or more other of the additional ingredients described herein. Other opthalmically-administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form or in a liposomal preparation.

[0197] As used herein, “additional ingredients” include, but are not limited to, one or more of the following: excipients; surface active agents; dispersing agents; inert diluents; granulating and disintegrating agents; binding agents; lubricating agents; sweetening agents; flavoring agents; coloring agents; preservatives; physiologically degradable compositions such as gelatin; aqueous vehicles and solvents; oily vehicles and solvents; suspending agents; dispersing or wetting agents; emulsifying agents, demulcents; buffers; salts; thickening agents; fillers; emulsifying agents; antioxidants; antibiotics; antifungal agents; stabilizing agents; and pharmaceutically acceptable polymeric or hydrophobic materials. Other “additional ingredients” which may be included in the pharmaceutical compositions of the invention are known in the art and described, for example in Genaro, ed., 1985, Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., which is incorporated herein by reference.

[0198] Therapeutically effective doses and dosing regimens

[0199] In some embodiments, the therapeutically effective dose of at least one of the disclosed peptides may be administered at least once, twice, three, or four times within a three-month period.

[0200] Therapeutically effective doses and dosing regimens of the foregoing methods may vary, as would be readily understood by those of skill in the art. Dosage regimens may be adjusted to provide the optimum desired response. For example, in some embodiments, a single dose of the peptide may be administered, while in some embodiments, several divided doses may be administered over time, or the dose may be proportionally reduced or increased in subsequent dosing as indicated by the situation. For example, in some embodiments the disclosed peptides may be administered once or twice weekly by subcutaneous, intravenous, or intramuscular injection. In some embodiments, the disclosed peptides may be administered once or twice monthly by subcutaneous, intravenous, or intramuscular injection. In some embodiments, the disclosed peptides thereof may be administered once or twice annually by subcutaneous, intravenous, or intramuscular injection. In some embodiments, the disclosed peptides may be administered once every week, once every other week, once every three weeks, once every four weeks, once every month, once every other month, once every three months, once every four months, once every five months, once every six months, once every seven months, once every eight months, once every nine months, once every ten months, once every eleven months, twice a year, or once a year, as the situation or condition of the patient may indicate.

[0201] The therapeutically effective dose of peptide administered to the patient (whether administered in a single does or multiple doses) should be sufficient to treat ischemic or toxic organ injury. Such therapeutically effective amount may be determined by evaluating the symptomatic changes in the patient.

[0202] Exemplary doses can vary according to the size and health of the individual being treated, as well as the condition being treated. In some embodiments, the effective amount of a disclosed peptide is about 2,200 mg; however, in some situations the dose may be higher or lower. In some embodiments, a therapeutically effective amount may be between 50 and 5000 mg, between 60 about 4500 mg, between 70 and 4000 mg, between 80 and 3500 mg, between 90 and 3000 mg, between 100 and 2500 mg, between 150 and 2000 mg, between 200 and 1500 mg, between 250 and 1000 mg, or any dose in between. For instance, in some embodiments, the therapeutically effective amount may be about 50 about 60, about 70 , about 80, about 90, about 100, about 150, about 200, about 250, about 300, about 350, about 400, about 450, about 500, about 550, about 600, about 650, about 700, about 750, about 800, about 850, about 900, about 950, about 1000, about 1100, about 1200, about 1300, about 1400, about 1500, about 1600, about 1700, about 1800, about 1900, about 2000, about 2100, about 2200, about 2300, about 2400, about 2500, about 2600, about 2700, about 2800, about 2900, about 3000, about 3100, about 3200, about 3300, about 3400, about 3500, about 3600, about 3700, about 3800, about 3900, about 4000, about 4100, about 4200, about 4300, about 4400, about 4500, about 4600, about 4700, about 4800, about 4900, about 5000 or more mg.

[0203] Similarly, in some embodiments, the effective amount of peptide is about 25 mg / kg; however, in some embodiments, the concentration may be higher or lower. In some embodiments, the effective amount may be about 1-50 mg / kg, about 5-40 mg / kg, about 10-30 mg / kg, or about 15-25 mg / kg or any value in between. For instance, in some embodiments, the effective amount may be 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, 26, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 or more mg / kg.

[0204] The disclosed methods of treatment may also be combined with other known methods of treatment as the situation may require.

[0205] Therapeutically effective doses and dosing regimens of the foregoing methods may vary, as would be readily understood by those of skill in the art. Dosage regimens may be adjusted to provide the optimum desired response. For example, in some embodiments, a single bolus dose of the peptide may be administered, while in some embodiments, several divided doses may be administered over time, or the dose may be proportionally reduced or increased in subsequent dosing as indicated by the situation. For example, in some embodiments the disclosed peptides may be administered once or twice weekly by subcutaneous, intravenous, or intramuscular injection. In some embodiments, the disclosed peptides may be administered once or twice monthly by subcutaneous, intravenous, or intramuscular injection. In some embodiments, the disclosed peptides may be administered once or twice annually by subcutaneous, intravenous, or intramuscular injection. In some embodiments, the disclosed peptides may be administered once every week, once every other week, once every three weeks, once every four weeks, once every month, once every other month, once every three months, once every four months, once every five months, once every six months, once every seven months, once every eight months, once every nine months, once every ten months, once every eleven months, twice a year, or once a year, as the situation or condition of the patient may indicate.

[0206] Typically dosages of the compound of the invention which may be administered to an animal, preferably a human, range in amount from about 0.01 mg to about 1000 mg per kilogram of body weight of the animal. The precise dosage administered will vary depending upon any number of factors, including, but not limited to, the type of animal and type of disease or disorder being treated, the age of the animal and the route of administration. Preferably, the dosage of the compound will vary from about 1 mg to about 100 mg per kilogram of body weight of the animal. The compound can be administered to an animal as frequently as several times daily, or it can be administered less frequently, such as once a day, once a week, once every two weeks, once a month, or even less frequently, such as once every several months or even once a year or less. The frequency of the dose will be readily apparent to the skilled artisan and will depend upon any number of factors, such as, but not limited to, the type and severity of the disease or disorder being treated, the type and age of the animal, etc.

[0207] EXPERIMENTAL EXAMPLES

[0208] The invention is further described in detail by reference to the following experimental examples. These examples are provided for purposes of illustration only, and are not intended to be limiting unless otherwise specified. Thus, the invention should in no way be construed as being limited to the following examples, but rather, should be construed to encompass any and all variations which become evident as a result of the teaching provided herein.

[0209] Without further description, it is believed that one of ordinary skill in the art can, using the preceding description and the following illustrative examples, make and utilize the present invention and practice the claimed methods. The following working examples therefore are not to be construed as limiting in any way the remainder of the disclosure.

[0210] Example 1 : Modified renalase agonists peptides display increased in vivo and / or in vitro half-life. Renalase agonist peptides are effective in treating and preventing ischemic and toxic organ injury. These short peptides have a short ’A life (10-20 min) and need to be administered frequently. These peptides were modified in a manner that increases ’A life from minutes to days. Serum albumin, which has a long Vi life, was linked via its carboxy terminal to renalase agonist peptides. This increased the renalase agonist peptides A life from minutes to days (Figures 3-8).

[0211] Figures 9-10: increased in vitro potency of palmitoylated RP10 peptide in protecting against cisplatin toxicity in cells in culture model.

[0212] The materials and methods employed in these experiments are now described.

[0213] Peptide synthesis

[0214] The peptides were synthesized on a ChemMatrix Rink Amide resin, using standard Fmoc- synthesis protocol on an APEX 396 automatic synthesizer. For removal of the Fmoc-protecting group: The resin was swollen in N,N-dimethylformamide (DMF) for 30 minutes, treated at 50°C with 20% piperidine-DMF for 8 minutes, and washed three times with DMF. For the coupling reaction: The Fmoc-protected amino acid, 6-chloro-l- hydroxybenzotriazole (CI-HOBt), diisodopropyl-carbodiimide (DCI) and Nmethyl- 2-pyrrolidine (NMP) were added to the resin. The mixture was vortexed for 20 minutes at 50°C. Afterwards, the resin was washed with DMF once. The cycle of Fmoc-deprotection and coupling steps was repeated until the last amino acid residue was assembled. After removal of the final Fmoc-protecting group, the resin was treated with 20% acetic anhydride-NMP for 20 minutes and was then washed with DMF, methylene chloride (DCM), and dried with air. The peptides were cleaved from the resin using a trifluoroacetic acid (TFA) cocktail [95% TFA, 2.5% water, and 2.5% triisopropyl silane (TIS)) for three hours. Crude peptides were precipitated by adding ice-chilled anhydrous ethyl ether, washed three times with anhydrous ethyl ether, and dried in vacuo.

[0215] Recombinant protein production

[0216] Recombinant proteins were produced via standard procedures known in the field. 4 recombinant proteins (SEQ ID NOs: 9-12) were produced and quantitated (Figure 1 and Figure 2). Coomassie blue staining (Figure 2B) validated the production of 4 different recombinant proteins (SEQ ID NOs: 9-12). Figure 2C, and Table 1, presents the quantitation, using standard protein quantitation procedures, of the recombinant proteins.

[0217] Table 1 : In vitro recombinant protein yield

[0218] In vivo recombinant protein kinetic studies

[0219] In vivo protein kinetic studies were performed by routine procedure known in the field. Specifically, 3 C75BL / 6 male mice (weighing approximately 25g each) were intravenously (IV) injected (retro-orbital) with 20mg of recombinant protein per kg of mouse (Figure 3). Approximately 30pl of serum was extracted from the mice at 15 minutes, 1 day, 3 days, 5 days, and 7 days post injection (Figure 3). Serum levels of recombinant protein were measured via ELISA sandwich (Figure 4).

[0220] ELISA for quantitation of recombinant proteins

[0221] ELISA was performed by standard procedures known in the field (Figure 4). Specifically, serum from mouse blood was extracted and incubated on standard ELISA plates coated with a-Strep antibody. Serum was incubated on the ELISA plates and recombinant protein bound to the oc-Strep antibody was detected with huM28 chimera antibody (detection antibody). Conjugated a-human IgG (secondary antibody) was bound to the detection antibody (huM28 chimera antibody) and subsequently exposed to substrate to enable detection of the secondary antibody and quantification of the recombinant protein. A representative standard curve was produced to enable quantitation of recombinant protein in vivo (Figure 5).

[0222] Recombinant proteins with modified renalase agonist peptides displayed increased halflife in vivo

[0223] Recombinant protein (SEQ ID NO: 11) displayed increased half-life in vivo following mouse IV retro-orbital injections (Figure 6 through Figure 8). 3 mice were injected with SEQ ID NO: 11 and protein kinetic studies were performed as described above. SEQ ID NO: 11 was determined to have an in vivo half life of 1-3 days at 20mg / kg (Figure 6 through Figure 8). Figure 7 depicts representative data demonstrating that linking peptide SEQ ID NO: 6 to mouse albumin (SEQ ID NO: 11) increases its halflife in mouse plasma from minutes to days. Protein kinetic experiments in Figure 7 demonstrate that SEQ ID NO: 11 has a half-life of approximately 2 days in vivo. Table 2 presents results of protein kinetic experiments carried out in 3 mice as presented in Figure 8. Increased half-life of the recombinant protein SEQ ID NO: 11 occurred through recycling via the neonatal Fc receptor.

[0224] Table 2: In vivo serum concentration of recombinant protein

[0225] Modified renalase agonist peptides displayed increased half-life in vitro

[0226] Modified renalase agonist peptide SEQ ID NOs: 13-16 (Figure 9) were produced according to standard procedures as described above. Conjugation of modified renalase agonist peptides to myristoleic acid or palmitoleic acid (SEQ ID NOs: 13-16) promoted binding to human serum albumin and increased half-life through recycling via the neonatal Fc receptor. In vitro cisplatin studies were performed to quantitate increase in potency of modified renalase agonist peptides. SEQ ID NO: 16 was determined to have increased potency compared to SEQ ID NO: 6 as determined by relative cell viability in in vitro cisplatin studies (Figure 10).

[0227] Modified renalase agonist peptide SEQ ID NO: 16 displays increased half-life in vivo Modified renalase agonist peptide SEQ ID NO: 16 is determined to have increased in vivo half-life compared to SEQ ID NO: 6. Modified renalase agonist peptide SEP ID NO: 16 promotes better mouse survival in vivo

[0228] Modified renalase agonist peptide SEQ ID NO: 16 is determined to promote prolonged survival of mice using a mouse model of renalase deficiency associated disease and / or disorder. Example 2: Sequences

[0229] Table 3: Amino acid sequences of the present invention

[0230] The disclosures of each and every patent, patent application, and publication cited herein are hereby incorporated herein by reference in their entirety. While this invention has been disclosed with reference to specific embodiments, it is apparent that other embodiments and variations of this invention may be devised by others skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

Claims

CLAIMSWhat is claimed is:

1. A modified renalase agonist peptide comprising at least one amino acid sequence that is at least 90% identical to the amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 6, 13, 14, 15, 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, and 36.

2. The modified renalase agonist peptide of claim 1, wherein the modified renalase agonist peptide comprises SEQ ID NOs: 5, 6, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, and 36, and further comprises at least one N- terminal glycine residue.

3. The modified renalase agonist peptide of claim 1 or 2, wherein the at least one modified renalase agonist peptide comprises at least one modification selected from the group consisting of addition of a myristoleic acid residue and addition of a palmitoleic acid residue, and wherein the modified renalase agonist peptide is amidated at the C-terminus.

4. The modified renalase agonist peptide of claim 3, wherein the addition of a myristoleic acid residue and addition of a palmitoleic acid residue is at the N’ terminus of the modified renalase agonist peptide.

5. The modified renalase agonist peptide of claim 4 wherein the modified renalase agonist peptide comprises SEQ ID NO: 13, 14, 15, or 16.

6. The modified renalase agonist peptide of any of claims 1-5, wherein the modified renalase agonist peptide further comprises at least one modification to increase half-life or stability of the modified renalase agonist peptide.

7. The modified renalase agonist peptide of claim 6, wherein the at least one modification is the addition of at least one D amino acid.

8. A fusion peptide comprising the modified renalase agonist peptide of any of claims 1-5, wherein the modified renalase agonist peptide is linked to an additional amino acid sequence or peptide.

9. The fusion peptide of claim 8, wherein the modified renalase agonist peptide is N-terminally linked to human serum albumin.

10. The fusion peptide of claim 9, wherein the human serum albumin is at least 90% identical to the amino acid sequence of SEQ ID NO: 17.

11. The fusion peptide of claim 8, comprising a linker between the modified renalase agonist peptide and the additional amino acid sequence or peptide.

12. The modified renalase agonist peptide or fusion peptide of any of claims 1-11, wherein the half-life of the modified renalase agonist peptide or fusion peptide is increased as compared to an unmodified renalase peptide.

13. The modified renalase agonist peptide of claim 2, wherein the at least one N-terminal glycine residue is linked to at least one molecule that enhances association of the peptide with human serum albumin.

14. A composition comprising the modified renalase agonist peptide or fusion peptide of any of claims 1-13.

15. The composition of claim 14, wherein the composition comprises at least one modified renalase agonist peptide and at least one fusion peptide, or at least two modified renalase agonist peptides, or at least two fusion peptides of any of claims 1-16. The composition of claim 14, wherein the composition is a pharmaceutical composition.

17. The composition of claim 16, further comprising at least one pharmacologically acceptable carrier.

18. A method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the modified renalase agonist peptide, fusion peptide, or composition of any of claims 1-17.

19. The method of claim 18, wherein the disease or disorder comprises a renal condition, a cardiovascular condition, a heart condition, a central nervous system (CNS) condition, or pre-eclampsia.

20. The method of claim 19, wherein the renal condition comprises end-stage renal disease (ESRD), chronic renal failure, and any combination thereof, wherein the cardiovascular conditions comprises hypertension, asymptomatic left ventricular dysfunction, chronic congestive heart failure (CHF), myocardial infarction (MI), cardiac rhythm disturbance, stroke, atherosclerosis, and any combination thereof, wherein the CNS condition comprises depression, anxiety, mania, schizophrenia, post- traumatic stress disorder (PTSD), and any combination thereof.

21. The method of claim 20, wherein the hypertension comprises chronic hypertension, systolic hypertension, isolated systolic hypertension, diabetic hypertension, pulmonary hypertension, acute severe hypertension, and any combination thereof,

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