Modified caveolin-1 peptides for the treatment of chronic kidney disease

Abstract

Provided herein are methods of using modified caveolin-1 (Cav-1) peptides to treat or prevent kidney diseases or disorders in a subject. In particular, provided are methods of using modified Cav-1 peptides for the treatment of chronic kidney diseases characterized by fibrosis, such as, for example, Alport syndrome.

Description

[Technical field]

[0001] The present disclosure generally relates to the field of molecular biology and medicine.In particular, the present disclosure provides a composition comprising modified caveolin-1 peptide for treating chronic kidney disease.The present disclosure also provides a composition comprising modified caveolin-1 peptide for treating fibrotic disease in elderly subjects.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 270,852, filed October 22, 2021, the contents of which are incorporated by reference in their entirety.

[0003] Reference to Electronic Sequence Listing The contents of the electronic sequence listing (LUTX_024_01WO_SeqList_ST26.xml, size: 162,055 bytes, and creation date: October 11, 2022) are incorporated herein by reference in their entirety. [Background technology]

[0004] Chronic kidney disease represents a global health problem affecting over 20 million Americans and approximately 10% of the world's population. In chronic kidney disease, progressive loss of renal function occurs as a result of the deposition of fibrous tissue between the functional units of the kidney or nephron, as well as continued replacement of fibrous tissue at the filtering surface. Renal fibrosis is the pathological hallmark of chronic kidney disease and is a major contributing factor in the progression to end-stage renal disease, where individuals require routine dialysis or kidney transplantation for survival.

[0005] The clinical management of chronic kidney disease mainly focuses on controlling blood pressure using renin-angiotensin system (RAS) inhibitors to slow disease progression, but this treatment is not curative.Therefore, there is still a need in the art for alternative therapies that target the fibrotic process of the kidney, which can slow or stop the progression of chronic kidney disease. Summary of the Invention

[0006] The present disclosure provides a method for treating or preventing a kidney disease or disorder in a subject, the method comprising administering to the subject an effective amount of a modified Cav-1 peptide, the modified Cav-1 peptide comprising: (a) any one of the amino acid sequences set forth in SEQ ID NOs: 2-111; (b) any one of the amino acid sequences set forth in SEQ ID NOs: 2-111; or (c) any one of the amino acid sequences set forth in SEQ ID NOs: 2-111 including one or more amino acid substitutions, insertions, deletions, or chemical modifications.

[0007] In some embodiments, the modified Cav-1 peptide comprises L-amino acids. In some embodiments, the modified Cav-1 peptide comprises D-amino acids. In some embodiments, the modified Cav-1 peptide comprises both L-amino acids and D-amino acids. In some embodiments, the modified Cav-1 peptide comprises deuterated residues. In some embodiments, the modified Cav-1 peptide comprises at least one non-standard amino acid. In some embodiments, the non-standard amino acid is ornithine.

[0008] In some embodiments, the modified Cav-1 peptide comprises an N-terminal modification. In some embodiments, the modified Cav-1 peptide comprises a C-terminal modification. In some embodiments, the modified Cav-1 peptide comprises an N-terminal modification and a C-terminal modification. In some embodiments, the N-terminal modification is acylation. In some embodiments, the C-terminal modification is amidation.

[0009] In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of FTTFTVT (SEQ ID NO: 3). In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of KASFTTFTVTKGS (SEQ ID NO: 4). In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of KKASFTTFTVTKGS-NH2 (SEQ ID NO: 5). In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of aaEGKASFTTFTVTKGSaa (SEQ ID NO: 6). In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 7). In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8). In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of OASFTTFTVTOS (SEQ ID NO: 9). In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of OASFTTFTVTOS-NH2 (SEQ ID NO: 10).

[0010] In some embodiments, the modified Cav-1 peptide comprises an internalization sequence. In some embodiments, the internalization sequence is located at the C-terminus of the peptide. In some embodiments, the internalization sequence is located at the N-terminus of the peptide.

[0011] In some embodiments, the modified Cav-1 peptide further comprises a cap at the N-terminus and / or C-terminus. In some embodiments, the modified Cav-1 peptide comprises a cap at the N-terminus and C-terminus.

[0012] In some embodiments, the modified Cav-1 peptide is cyclized.

[0013] In some embodiments, the modified Cav-1 peptide maintains the biological activity of native Cav-1 (SEQ ID NO:1).

[0014] In some embodiments, the modified Cav-1 peptide is a multimer comprising at least two peptides as described herein. In some embodiments, a first peptide of the at least two peptides is essentially identical to a second peptide of the at least two peptides. In some embodiments, a first peptide of the at least two peptides is not identical to a second peptide of the at least two peptides.

[0015] In some embodiments, the modified Cav-1 peptide is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, or orally. In some embodiments, the modified Cav-1 peptide is administered subcutaneously. In some embodiments, the modified Cav-1 peptide is administered intravenously.

[0016] In some embodiments, the kidney disease or disorder is selected from the group consisting of chronic kidney disease, end-stage renal disease, glomerulonephritis, focal segmental glomerulosclerosis, renal fibrosis, polycystic kidney disease, IgA nephropathy, lupus nephritis, nephrotic syndrome, Alport syndrome, amyloidosis, Goodpasture syndrome, granulomatosis with polyangiitis, or acute kidney injury.In some embodiments, the kidney disease or disorder is Alport syndrome.In some embodiments, the kidney disease or disorder is characterized by fibrosis.

[0017] In some embodiments, the method further comprises administering an effective amount of at least one additional therapeutic agent. In some embodiments, the at least one additional therapeutic agent is an angiotensin-converting enzyme (ACE) inhibitor and / or an angiotensin II receptor (ARB) inhibitor. In some embodiments, the method further comprises treating the subject with dialysis.

[0018] In some embodiments, the modified Cav-1 peptide is administered as a composition comprising the modified Cav-1 peptide and at least one pharma- ceutically acceptable carrier or excipient.

[0019] A method of treating or preventing a fibrotic disease or disorder in an elderly subject, comprising administering to the subject an effective amount of a modified Cav-1 peptide comprising: (a) any one of the amino acid sequences of SEQ ID NOs: 2-111; (b) any one of the amino acid sequences of SEQ ID NOs: 2-111; or (c) any one of the amino acid sequences of SEQ ID NOs: 2-111, including one or more amino acid substitutions, insertions, deletions, or chemical modifications. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of FTTFTVT (SEQ ID NO: 3). In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8). In some embodiments, the fibrotic disease or disorder is interstitial lung disease. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0020] In some embodiments, the modified Cav-1 peptide is administered to the lungs. In some embodiments, the modified Cav-1 peptide is administered to the lungs via inhalation. In some embodiments, the modified Cav-1 peptide is administered to the subject using a nebulizer. In some embodiments, the modified Cav-1 peptide is administered to the subject using an inhaler.

[0021] In some embodiments, the modified Cav-1 peptide is formulated for inhalation. In some embodiments, the modified Cav-1 peptide is formulated for pressurized metered dose inhalation. In some embodiments, the modified Cav-1 peptide is formulated as a dry powder. In some embodiments, the dry powder comprising the modified Cav-1 peptide is essentially excipient-free. In some embodiments, the dry powder is produced by a spray drying process, air jet milling, ball milling, or wet milling. In some embodiments, the modified Cav-1 peptide is formulated for nebulization.

[0022] In some embodiments, the method further comprises administering to the subject a therapeutically effective amount of at least one additional therapeutic agent, hi some embodiments, the at least one additional therapeutic agent is chloroquine, hydroxychloroquine, remdesivir, favipiravir, lopinavir, or ritonavir.

[0023] In some embodiments, the subject is at least 55 years old, at least 60 years old, at least 65 years old, at least 70 years old, at least 75 years old, at least 80 years old, at least 85 years old, or at least 90 years old. [Brief description of the drawings]

[0024] [Figure 1A] FIG. 1A shows a gel of urinary proteins from cohort 1 and cohort 2 Col4a3− / − mice prior to treatment with saline or APi2355 peptide. Urine was loaded onto the gel using creatinine levels as a calibration standard. Cohort 1 began treatment at 6 weeks of age and cohort 2 at 5 weeks of age. Mouse IDs 1046 and 1117 died during treatment for unknown reasons. Asterisks indicate mouse albumin.

[0025] [Figure 1B] FIG 1B shows a gel of urinary proteins from cohort 1 and cohort 2 Col4a3- / - mice after daily intraperitoneal injections of APi2355 peptide or saline for 2 weeks. Cohort 1 began treatment at 6 weeks of age and cohort 2 at 5 weeks of age. Asterisks indicate mouse albumin.

[0026] [Figure 1C] Figure 1C shows the urinary albumin to creatinine ratio (g / mg) of Cohort 1 (left panel) and Cohort 2 (right panel) Col4a3- / - mice before and after 2 weeks of treatment with saline or APi2355 peptide. Cohort 1 began treatment at 6 weeks of age, and Cohort 2 began treatment at 5 weeks of age.

[0027] [Diagram 2] Figure 2 shows blood urea nitrogen (BUN) levels in Cohort 1 (left panel) and Cohort 2 (right panel) Col4a3- / - mice after treatment with saline or APi2355 peptide. Cohort 1 began treatment at 6 weeks of age, and Cohort 2 began treatment at 5 weeks of age.

[0028] [Figure 3A] Figure 3A shows immunofluorescence staining of collagen I and nidogen on kidney tissue from untreated Col4a3+ / - mice (left panel) or Col4a3- / - mice after 2 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel). Treatment began at 6 weeks of age.

[0029] [Figure 3B] Figure 3B shows immunofluorescence staining of collagen I and nidogen on kidney tissue from untreated Col4a3+ / - mice (left panel) or Col4a3- / - mice after 2 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel). Treatment began at 6 weeks of age.

[0030] [Figure 4A] Figure 4A shows immunofluorescence staining of collagen I and alpha-smooth muscle actin (αSMA) on kidney tissue from untreated Col4a3+ / − mice (left panel) or Col4a3− / − mice after 2 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel). Treatment began at 6 weeks of age.

[0031] [Figure 4B] Figure 4B shows immunofluorescence staining of collagen I and alpha-smooth muscle actin (αSMA) on kidney tissue from untreated Col4a3+ / − mice (left panel) or Col4a3− / − mice after 2 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel). Treatment began at 6 weeks of age.

[0032] [Figure 5A] Figure 5A shows immunofluorescence staining of collagen I and nidogen on kidney tissue from untreated Col4a3+ / - mice (left panel) or Col4a3- / - mice after 2 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel). Treatment began at 5 weeks of age.

[0033] [Figure 5B] Figure 5B shows immunofluorescence staining of collagen I and nidogen on kidney tissue from untreated Col4a3+ / - mice (left panel) or Col4a3- / - mice after 2 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel). Treatment began at 5 weeks of age.

[0034] [Figure 6A] Figure 6A shows immunofluorescence staining of collagen I and alpha-smooth muscle actin (αSMA) on kidney tissue from untreated Col4a3+ / − mice (left panel) or Col4a3− / − mice after 2 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel). Treatment began at 5 weeks of age.

[0035] [Figure 6B] Figure 6B shows immunofluorescence staining of collagen I and alpha-smooth muscle actin (αSMA) on kidney tissue from untreated Col4a3+ / − mice (left panel) or Col4a3− / − mice after 2 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel). Treatment began at 5 weeks of age.

[0036] [Figure 7A] Figure 7A shows a gel of urinary proteins from 4-week-old Col4a3- / - mice before treatment with saline or APi2355 peptide. Urine was loaded onto the gel using creatinine levels as a calibration standard. Asterisks indicate mouse albumin.

[0037] [Figure 7B] Figure 7B shows urinary protein gels from 6-week-old female (F) and male (M) Col4a3- / - mice after daily intraperitoneal injections of APi2355 peptide or saline for 2 weeks. Asterisks indicate mouse albumin.

[0038] [Figure 7C] Figure 7C shows urinary protein gels from 7- and 8-week-old female (F) and male (M) Col4a3- / - mice after daily intraperitoneal injections of APi2355 peptide or saline for 3 or 4 weeks. Asterisks indicate mouse albumin.

[0039] [Figure 7D] FIG. 7D shows the urinary albumin to creatinine ratio (g / mg) of Col4a3− / − mice before treatment at 4 weeks of age and during treatment with saline or APi2355 peptide at 6, 7, and 8 weeks of age.

[0040] [Figure 8] FIG. 8 shows blood urea nitrogen (BUN) levels in Col4a3− / − mice before treatment at 4 weeks of age and during treatment with saline or APi2355 peptide at 6 and 8 weeks of age.

[0041] [Figure 9A] FIG. 9A shows immunofluorescence staining of collagen I and nidogen on kidney tissue obtained from untreated male Col4a3+ / − mice.

[0042] [Figure 9B] FIG. 9B shows immunofluorescence staining of collagen I and nidogen on kidney tissue from female Col4a3− / − mice treated daily with APi2355 peptide for 4 weeks.

[0043] [Figure 9C]FIG. 9C shows immunofluorescence staining of collagen I and nidogen on kidney tissue from female Col4a3− / − mice treated daily with saline for 4 weeks.

[0044] [Figure 9D] FIG. 9D shows immunofluorescence staining of collagen I and nidogen on kidney tissue from female Col4a3− / − mice treated daily with APi2355 peptide for 4 weeks.

[0045] [Figure 9E] FIG. 9E shows immunofluorescence staining of collagen I and nidogen on kidney tissue from female Col4a3− / − mice treated daily with APi2355 peptide for 4 weeks.

[0046] [Figure 9F] FIG. 9F shows immunofluorescence staining of collagen I and nidogen on kidney tissue from female Col4a3− / − mice treated daily with saline for 4 weeks.

[0047] [Figure 9G] FIG. 9G shows immunofluorescence staining of collagen I and nidogen on kidney tissue from male Col4a3− / − mice treated daily with APi2355 peptide for 4 weeks.

[0048] [Figure 9H] FIG. 9H shows immunofluorescence staining of collagen I and nidogen on kidney tissue from male Col4a3− / − mice treated daily with APi2355 peptide for 4 weeks.

[0049] [Figure 10A]FIG. 10A shows immunofluorescence staining of caveolin-1 and laminin-111 (LM111) on intestinal tissue from untreated Col4a3+ / − mice (left panel) or Col4a3− / − mice after 4 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel).

[0050] [Figure 10B] FIG. 10B shows immunofluorescence staining of caveolin-1 and laminin-111 (LM111) on intestinal tissue from untreated Col4a3+ / − mice (left panel) or Col4a3− / − mice after 4 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel).

[0051] [Figure 10C] FIG. 10C shows immunofluorescence staining of caveolin-1 and laminin-111 (LM111) on kidney tissue from untreated Col4a3+ / − mice (left panel) or Col4a3− / − mice after 4 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel).

[0052] [Figure 10D] FIG. 10D shows immunofluorescence staining of caveolin-1 and laminin-111 (LM111) on kidney tissue from untreated Col4a3+ / − mice (left panel) or Col4a3− / − mice after 4 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel).

[0053] [Figure 10E] FIG. 10E shows immunofluorescence staining of caveolin-1 and laminin-111 (LM111) on kidney tissue from untreated Col4a3+ / − mice (left panel) or Col4a3− / − mice after 4 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel).

[0054] [Figure 10F]FIG. 10F shows immunofluorescence staining of caveolin-1 and laminin-111 (LM111) on kidney tissue from untreated Col4a3+ / − mice (left panel) or Col4a3− / − mice after 4 weeks of treatment with saline (middle panel) or APi2355 peptide (right panel).

[0055] [Figure 11A] FIG. 11A shows immunofluorescence staining of caveolin-1, collagen I, laminin-111, and nidogen on kidney tissue from untreated Col4a3+ / − and Col4a3− / − mice, or Col4a3− / − mice after 4 weeks of treatment with saline or APi2355 peptide.

[0056] [Figure 11B] FIG. 11B shows quantification of glomerular fibrosis in 8-week-old female Col4a3− / − mice after 4 weeks of treatment with saline or APi2355 peptide.

[0057] [Figure 12] FIG. 12 shows collagen in total lung homogenates from saline (SAL)- or bleomycin (BLM)-treated aged mice administered control peptide (CP) or CSP-7 via dry powder inhalation (DPI).

[0058] [Figure 13] Figure 13 shows collagen in total lung homogenates of saline (SAL) or bleomycin (BLM) treated aged mice administered control peptide (CP) or CSP-7 via dry powder inhalation (DPI). ****P<0.0001.

[0059] [Figure 14A]FIG. 14A shows total SMADs (tSMADs, upper panel) and phosphorylated SMADs (pSMADs, lower panel) in total lung homogenates of bleomycin (BLM)-treated aged mice that were untreated, or administered a control peptide (CP) or CSP-7 by dry powder inhalation, or CSP-7 by intraperitoneal injection (IP).

[0060] [Figure 14B] FIG. 14B shows galectin 7 in total lung homogenates of bleomycin (BLM)-treated aged mice that were untreated, or administered a control peptide (CP) or CSP-7 by dry powder inhalation, or administered CSP-7 by intraperitoneal injection (IP).

[0061] [Figure 15A] Figure 15A shows immunohistochemical staining of caveolin-1 (Cav-1) at 5x and 63x magnification in renal tissues obtained from normal donor A and normal donor B. Arrows indicate positive Cav-1 staining in glomeruli of normal renal tissue.

[0062] [Figure 15B] Figure 15B shows immunohistochemical staining of caveolin-1 (Cav-1) at 5x and 63x magnification in fibrotic renal tissue from Patient C, Patient D, and Patient E. Arrows indicate glomeruli within the fibrotic renal tissue. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0063] The present disclosure provides modified caveolin-1 (Cav-1) peptides and their uses for treating or preventing chronic kidney disease in a subject. Also provided herein is a method for treating fibrotic diseases or disorders in an elderly subject by administering modified Cav-1 peptides or pharmaceutical compositions thereof.

[0064] I. Definition As used herein, the article "a" or "an" refers to one or more of the grammatical objects of the article. As used herein in the claims, when used in conjunction with the word "comprising," the article "a" or "an" refers to one or more of the grammatical objects of the article.

[0065] Use of the term "or" in the claims is used to mean "and / or" unless expressly indicated to refer only to alternatives or the alternatives are not mutually exclusive, although the present disclosure supports the definition to refer only to alternatives and "and / or." As used herein, "another" can mean at least a second, or more.

[0066] Throughout this application, the term "about" is used to indicate that a value includes the inherent variation of error for the device or method used to determine the value or the variation that exists between samples being measured. Unless otherwise stated or apparent from the context, the term "about" means within a range of 10% above or below the reported numerical value (except where such numerical value exceeds 100% or falls below 0% of possible values). When used in conjunction with a range or series of values, the term "about" applies to the endpoints of the range or each recited numerical value in the series, unless otherwise indicated. As used in this application, the terms "about" and "approximately" are used as equivalents.

[0067] The terms "peptide," "polypeptide," or "protein" are used in their broadest sense to refer to a molecule of two or more amino acids, amino acid analogs, or peptidomimetics. In some embodiments, the amino acids are linked by peptide bonds. In some embodiments, the amino acids are linked by other types of bonds, e.g., esters, ethers, etc. As used herein, the term "amino acid" refers to either natural and / or unnatural or synthetic amino acids, including glycine and both D or L optical isomers, amino acid analogs, and peptidomimetics.

[0068] The term "peptidomimetic" or "peptide mimetic" refers to a peptide that has been modified to include at least one non-peptide bond, such as, for example, a urea bond, a carbamate bond, a sulfonamide bond, a hydrazine bond, or any other covalent bond.

[0069] In some embodiments, a polypeptide or polypeptide has a certain percentage (e.g., 80%, 85%, 90%, or 95%) of "sequence identity" or "homology" to another sequence, meaning that when aligned, the percentage of amino acids are the same in comparing the two sequences. In some embodiments, the term "identity" or "homology" refers to the percentage of amino acid residues in a candidate sequence that are identical to the residues of the corresponding sequence, and the candidate sequence is compared to the corresponding sequence after aligning the sequences and introducing gaps as necessary to achieve the maximum percentage identity throughout the sequence. Alignment and percentage of homology or sequence identity can be determined using software programs known in the art, such as those described in Current Protocols In Molecular Biology (FMA Usubel et al., eds., 1987) Supplement 30, section 7.7.18, Table 7.7.1.

[0070] The term "substantially pure" refers to a peptide or polypeptide that has been isolated and purified to at least some degree from components that naturally accompany it. Typically, a peptide or polypeptide is substantially pure when it is at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99% by weight free from the proteins and naturally occurring organic molecules with which it is naturally associated. For example, a substantially pure peptide or polypeptide can be obtained by extraction from a natural source, by expressing a recombinant nucleic acid in a cell that does not normally express the protein, or by chemical synthesis.

[0071] As used herein, the term "isolated" refers to a peptide or polypeptide that has been separated from any natural environment, such as a bodily fluid, e.g., blood, and that has been separated from components that naturally accompany the peptide.

[0072] As used herein, "essentially free" with respect to a specified component means that none of the specified components are intentionally formulated into the composition and / or are present only as contaminants or trace amounts. Thus, the total amount of the specified component resulting from any unintentional contamination of the composition is well below 0.01%. Most preferred are compositions that have no detectable amount of the specified component using standard analytical methods.

[0073] The term "variant" as used herein refers to a polypeptide or nucleic acid that differs from a reference polypeptide or nucleic acid by one or more amino acid or nucleic acid deletions, additions, substitutions, or side chain modifications, but retains one or more specific functions or biological activities of the native molecule. Also included within the term variant are polynucleotides or polypeptides that may be altered in primary, secondary, or tertiary structure compared to a reference polynucleotide or polypeptide, respectively (e.g., compared to a wild-type polynucleotide or polypeptide).

[0074] The term "insertion" refers to the addition of one or more amino acids in a peptide or polypeptide sequence, and the term "deletion" refers to the removal of one or more amino acids in a peptide or polypeptide sequence. "Insertions" and "deletions" are typically in the range of about 1-5 amino acids. Mutations can be determined experimentally by synthetically producing the peptide using recombinant DNA technology to systematically create insertions, deletions, or substitutions of nucleotides in the sequence.

[0075] The term "substitution" when referring to a peptide or polypeptide refers to a change of an amino acid to a different entity, e.g., another amino acid or amino acid moiety. Amino acid substitutions include changes in which an amino acid is replaced with a different natural amino acid residue or a non-conventional amino acid residue. Such substitutions may be classified as "conservative", where an amino acid residue contained in a polypeptide is replaced with another natural amino acid with similar characteristics, either in terms of polarity, side chain functionality, or size. Such conservative substitutions are well known in the art. Substitutions encompassed by the present invention may also be "non-conservative", where an amino acid residue present in a peptide is replaced with an amino acid with different properties, such as a natural amino acid from a different group (e.g., replacing a charged or hydrophobic amino acid with alanine), or a natural amino acid is replaced with a non-conventional amino acid. In some embodiments, the amino acid substitutions are conservative. In some embodiments, the amino acid substitutions are non-conservative.

[0076] An "analog" refers to a molecule, such as a peptide, that is similar in function to either the entire molecule or a fragment thereof. The term "analog" is also intended to include allelic species and induced variants. Analogs typically differ from the native peptide at one or more amino acid residues, often by conservative amino acid substitutions. Analogs typically exhibit at least about 80% or at least about 90% sequence identity with the native peptide. Some analogs also include unnatural amino acids or modifications of the N- or C-terminal amino acids. Examples of unnatural amino acids include, but are not limited to, disubstituted amino acids, N-alkyl amino acids, lactate, 4-hydroxyproline, γ-carboxyglutamate, ε-Ν,Ν,Ν-trimethyllysine, ε-Ν-acetyllysine, O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylysine, and σ-Ν-methylarginine. Fragments and analogs can be screened for prophylactic or therapeutic efficacy using in vitro and in vivo methods well known to those of skill in the art.

[0077] The term "covalently linked" refers to a peptide or polypeptide that is directly or indirectly (e.g., via a linker) linked by a covalent chemical bond. In some embodiments, the fusion peptides of the present disclosure are covalently linked.

[0078] As used herein, the term "fusion protein" refers to a recombinant protein of two or more proteins. Fusion proteins can be produced, for example, by joining a nucleic acid sequence encoding one protein to a nucleic acid encoding another protein to form a single open reading frame that can be translated into a single polypeptide carrying all the intended proteins in a host cell. The order of protein arrangement can vary. Fusion proteins can include epitope tags or half-life extenders. Epitope tags include biotin, FLAG tag, c-myc, hemagglutinin, His6, digoxigenin, FITC, Cy3, Cy5, green fluorescent protein, V5 epitope tag, GST, β-galactosidase, AU1, AU5, and avidin. Half-life extenders include Fc domain and serum albumin.

[0079] The term "airway" as used herein refers to any part of the respiratory tract, including the upper airway, airways, and lungs. The upper respiratory tract includes the nose and nasal passages, the mouth, and the throat. The respiratory tract includes the larynx, trachea, bronchi, and bronchioles. The lungs include the respiratory bronchioles, alveolar ducts, alveolar sacs, and alveoli.

[0080] As used herein, the terms "atomization," "atomized," and other grammatical variations refer to the process of converting a liquid into small aerosol droplets using a nebulizer.

[0081] The term "air jet mill" refers to a device or method for reducing particle size by using a jet of compressed gas to collide particles with each other, thereby breaking them up. In some embodiments, an air jet mill is used to reduce the size of peptide particles. Other mechanical grinding devices that perform the same function can also be used interchangeably with an air jet mill. Air jet milling can occur under a variety of environmental parameters, such as temperature, pressure, relative / absolute humidity, oxygen content, etc.

[0082] The term "ball mill" refers to a device or method for reducing particle size by adding particles of interest and grinding media to the inside of a cylinder and rotating the cylinder. As the grinding media rises and falls along the outside of the cylinder while rotating, the particles of interest are broken down. In some embodiments, a ball mill is used to reduce the size of peptide particles. Other mechanical grinding devices that perform the same function can also be used interchangeably with the air jet mill.

[0083] The term "wet mill" or "media mill" refers to a device or method for reducing particle size by adding target particles to the device using an agitator containing a liquid and a media that includes grinding media. Upon addition of the target particles, as the agitator rotates, the energy dissipated by the agitator brings the grinding media into contact with the target particles, resulting in the breakage of the target particles. Other mechanical comminuting devices that perform the same function may be used interchangeably with the air jet mill.

[0084] The term "high pressure homogenization" refers to a method of reducing particle size by subjecting the target particles to a device that combines both pressure and mechanical forces to break them down. The mechanical forces used in high pressure homogenization can include impact, shear, and cavitation, among others. Other mechanical comminuting devices that perform the same function can also be used interchangeably with the air jet mill.

[0085] The term "pyrogenic grinder" refers to a device or method for reducing particle size by first cooling the target particles with dry ice, liquid nitrogen, or other cryogenic liquid, and then grinding the target particles to reduce their size. Other mechanical grinding devices that perform the same function may also be used interchangeably with air jet mills.

[0086] The term "effective amount" or "therapeutically effective amount" refers to an amount effective for treating and / or preventing a disease or disorder disclosed herein. In some embodiments, an effective amount is an amount or dose of a composition (e.g., a pharmaceutical composition, a compound, or a drug) that produces at least one desired therapeutic effect in a subject, such as preventing or treating a target condition or beneficially alleviating a symptom associated with the condition. The most desirable effective amount is an amount that produces the desired efficacy of a particular treatment selected by a person skilled in the art for a given subject in need thereof. This amount will vary depending on a variety of factors understood by a person skilled in the art, including, but not limited to, the characteristics of the composition (including activity, pharmacokinetics, pharmacodynamics, and bioavailability), the physiological condition of the subject (including age, sex, disease type, disease stage, general health, response to a given dose, and type of drug), the nature of the pharmaceutically acceptable carrier or carriers in the formulation, and the route of administration.

[0087] The term "pharmaceutical composition" or "pharmaceutical acceptable composition" refers to a composition that does not cause adverse, allergic, or other untoward reactions when administered to a subject. The preparation of pharmaceutical compositions containing modified Cav-1 peptides, such as CSP-7, or additional active ingredients will be known to those skilled in the art. Furthermore, for animal (e.g., human) administration, it will be understood that the preparation should meet the bioburden, sterility, pyrogenicity, general safety, and / or purity standards required by the FDA or other recognized regulatory authorities.

[0088] As used herein, the term "pharmaceutical acceptable carrier" includes any and all of excipients, processing aids, aqueous solvents (e.g., water, alcoholic / aqueous solutions, saline, parenteral vehicles such as sodium chloride, Ringer's dextrose, etc.), non-aqueous solvents (e.g., propylene glycol, polyethylene glycol, vegetable oils, and injectable organic esters such as ethyl oleate, etc.), dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial or antifungal agents, antioxidants, chelating agents, and inert gases), isotonicity agents, absorption retardants, salts, drugs, drug stabilizers, gels, binders, disintegrants, lubricants, flavor adjusters (e.g., sweeteners, flavoring agents), similar materials, and combinations thereof, as known to those skilled in the art. The pH and exact concentration of various components in the pharmaceutical composition are adjusted according to well-known parameters. In some embodiments, the carrier may encapsulate a therapeutic agent but may not itself be consumed or administered to a subject (e.g., a shell capsule surrounding a dry powder composition, such as for use in a dry powder inhaler). See, e.g., Remington's Pharmaceutical Sciences, 18th Ed., 1990, incorporated herein by reference.

[0089] As used herein, "excipient" refers to a pharma- ceutically acceptable carrier that is a relatively inert substance used to facilitate administration or delivery of an active pharmaceutical ingredient (API) (e.g., modified Cav-1 peptide) to a subject or to facilitate processing of the API into a drug formulation that can be used medicamentously for delivery to a site of action in a subject. Excipients or pharma-ceutically acceptable carriers include all of the inactive components of a dosage form, except for the active ingredient. Non-limiting examples of excipients include carrier agents, bulking agents, stabilizers, surfactants, surface modifiers, solubility enhancers, buffers, encapsulating agents, antioxidants, preservatives, non-ionic wetting or clarifying agents, viscosity enhancers, and absorption enhancers. The term "excipient-free" refers to the modified Cav-1 peptide or pharmaceutical composition thereof being in a formulation that does not include any excipients.

[0090] A "biologically active" caveolin-1 (Cav-1) peptide refers to a peptide that increases p53 protein levels, reduces urokinase plasminogen activator (uPA) and uPA receptor (uPAR), and / or increases plasminogen activator inhibitor-1 (PAI-1) expression in cells, such as fibrotic fibroblasts. In some embodiments, a biologically active peptide has at least 20% of the biological or biochemical activity (e.g., as measured by in vitro or in vivo assays) of the native Cav-1 polypeptide of SEQ ID NO:1. In some embodiments, a biologically active peptide has increased biological or biochemical activity compared to the native Cav-1 polypeptide.

[0091] The terms "subject", "individual" and "patient" are used interchangeably herein and refer to an animal, such as a human or a non-human animal (e.g., a mammal), to which treatment, including prophylactic treatment, with the modified Cav-1 peptide or pharmaceutical composition thereof disclosed herein is provided. As used herein, the term "subject" refers to a human and a non-human animal. The term "non-human animal" includes all vertebrates, such as mammals, such as non-human primates (especially higher primates), sheep, dogs, rodents (e.g., mice or rats), guinea pigs, goats, pigs, cats, rabbits, cows, and non-mammals, such as chickens, amphibians, reptiles, and the like. In some embodiments, the subject is a human. In some embodiments, the subject is an experimental animal or an animal substituted as a disease model. Non-human mammals include mammals, such as non-human primates (especially higher primates), sheep, dogs, rodents (e.g., mice or rats), guinea pigs, goats, pigs, cats, rabbits, and cows. In some embodiments, the non-human animal is a companion animal, such as a dog or a cat.

[0092] As used herein, the terms "treat", "treating" or "treatment" and grammatical variations thereof have the same meaning as commonly understood by those skilled in the art. In some embodiments, these terms may refer to an approach to obtain beneficial or desired clinical results. The terms may refer to delaying the onset or rate of onset of a condition, disorder, or disease, reducing or alleviating symptoms associated therewith, causing complete or partial regression of the condition, or any combination of any of the above. For purposes of the present invention, beneficial or desired clinical results include, but are not limited to, reduction or alleviation of symptoms, whether detectable or undetectable, reduction in the extent of the disease, stabilization of the disease state (e.g., not worsening), delay or slowing of disease progression, improvement or palliation of the disease state, and remission (whether partial or total). "Treat", "treating" or "treatment" may also mean prolonging survival compared to the expected survival if not treated. Thus, a subject (e.g., a human) in need of treatment may be a subject already suffering from the disease or disorder in question. The terms "treat," "treating," or "treatment" include the inhibition or reduction of the increase in severity of a pathology or symptom relative to the absence of treatment and are not necessarily meant to imply a complete cessation of the associated disease or condition.

[0093] As used herein, the terms "prevent", "preventing", "prevention" and their grammatical variations refer to an approach to prevent the onset or alter the pathology of a condition or disease. Thus, "prevention" can refer to a prophylactic measure. For the purposes of the present invention, beneficial or desired clinical results include, but are not limited to, the prevention or slowing down of symptoms, progression, or onset of a disease, whether detectable or undetectable. Thus, a subject (e.g., a human) in need of prevention can be a subject who is not yet afflicted with the disease or disorder in question. The term "prevention" includes delaying the onset of a disease compared to the absence of treatment, and is not necessarily meant to imply a permanent prevention of the associated disease, disorder, or condition. Thus, in certain circumstances, "preventing" or "prevention" of a condition can refer to reducing the risk of developing the condition, or preventing or delaying the onset of symptoms associated with the condition.

[0094] II. Caveolin-1 Peptides Embodiments of the present disclosure provide modified forms of native caveolin-1 (Cav-1) proteins, including but not limited to fragments, derivatives, and variants of native Cav-1 proteins. In some embodiments, the modified Cav-1 peptides are truncations of native Cav-1 polypeptides, such as the exemplary peptides shown in Table 2 and / or Table 3.

[0095] Native human Cav-1 is 178 amino acids long (see SEQ ID NO: 1 in Table 1 below) and has a molecular weight of 22 kDa. Caveolin-1 is an integral membrane protein involved in endocytosis, extracellular matrix organization, cholesterol distribution, cell migration, and signal transduction. See Boscher and Nabi, Adv Exp Med Biol, 2012;729-29-50. [Table 1]

[0096] In some embodiments, the modified Cav-1 peptide is the Cav-1 scaffolding domain (CSD). The CSD is composed of amino acids 82-101 of caveolin-1 (see SEQ ID NO:2 in Table 1 above). The CSD of caveolin-1 plays an important role in regulating caveolin-1 dimerization as well as various signaling intermediates (Shetty et al., Am J Respir Cell Mol Biol 2012;47:474-83; Fridolfsson et al., FASEB J 2014;28:3823-31; Degryse et al., Am J Physiol Cell Mol Physiol 2010;299:L442-L452; and Egger et al., PLos One,2013;8:e63432). The CSD domain of caveolin-1 has been shown to inhibit Wnt signaling, β-catenin-mediated transcription, and activation of SRC, EGFR, MEK1, and ERK-2 as well as various other factors (Shetty et al., Am J Respir Cell Mol Biol 2012;47:474-83; Bhandary et al., Am J Physiol Cell Mol Physiol 2012;302:L463-L473; Bhandary et al., Am J Pathol 2013;183:131-143; Fridolfsson et al., FASEB J 2014;28:3823-31; Degryse et al., Am J Physiol Cell Mol Physiol 2010;299:L442-L452; and Fiddler et al., Ann Am Thorac Soc, 2016;13:1430-2). For example, the CSD of Cav-1 interferes with Cav-1 interaction with SRC kinase and mimics the combined effects of uPA and anti-β1-integrin antibodies. The endogenous CSD domain can form homodimers with other Cav-1 proteins and interact with proteins that have caveolin-binding domain sequence (CBD) motifs.Up to 30% of all endogenous proteins are estimated to contain a CBD motif, and the caveolin-1 CSD domain is hypothesized to confer stability to these proteins (see Maruudamuthu et al., Am J Pathol 2015;185:55-68).

[0097] In some embodiments, the modified Cav-1 peptide is CSP-7, which is a seven amino acid fragment of the human CSD of caveolin-1 (see SEQ ID NO: 3 in Table 3 below).

[0098] Exemplary amino acid sequences of modified Cav-1 peptides are shown below in Tables 2 and 3. Capital letters indicate L-amino acids and lower case letters indicate D-amino acids (e.g., lower case "a" represents D-alanine). The term "Ac" refers to an acetyl group and the term "NH2" refers to an amide group. "O" refers to ornithine. [Table 2] [Table 3-1] [Table 3-2]

[0099] In some embodiments, the Cav-1 peptide or modified Cav-1 peptide is (a) consisting of any one of the amino acid sequences of SEQ ID NOs: 2 to 111; or (b) comprising any one of the core sequences of the amino acid sequences of SEQ ID NOs: 2 to 111; or (c) It comprises a core sequence of any one of the amino acid sequences of SEQ ID NOs: 2 to 111, and the core sequence comprises one or more amino acid substitutions, insertions, deletions, or chemical modifications.

[0100] In some embodiments, the Cav-1 peptide or modified Cav-1 peptide comprises a core sequence of any one of the amino acid sequences of SEQ ID NOs: 2-111. In some embodiments, the Cav-1 peptide or modified Cav-1 peptide comprises a core sequence of any one of the amino acid sequences of SEQ ID NOs: 2-111, wherein the core sequence comprises one or more amino acid substitutions, insertions, deletions, or chemical modifications. In some embodiments, the core sequence is SEQ ID NO: 3. In some embodiments, the core sequence is SEQ ID NO: 6.

[0101] In some embodiments, the Cav-1 polypeptide comprises or consists of the amino acid sequence of SEQ ID NO: 1. In some embodiments, the Cav-1 polypeptide comprises an amino acid sequence having at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 1. In some embodiments, the Cav-1 polypeptide comprises the amino acid sequence of SEQ ID NO: 1 with one or more mutations thereto. For example, in some embodiments, the Cav-1 polypeptide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations relative to SEQ ID NO: 1. In some embodiments, the Cav-1 polypeptide comprises the amino acid sequence of SEQ ID NO: 1 with 1-5, 5-10, 11-5, 15-20, 10-25, 25-30, or more than 30 mutations.

[0102] In some embodiments, the modified Cav-1 peptide comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide comprises an amino acid sequence having at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of any one of SEQ ID NOs: 2-111 with one or more mutations thereto. For example, in some embodiments, the modified Cav-1 peptide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations thereto. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of any one of SEQ ID NOs: 2-111 with 1-5, 5-10, or 11-15, or more mutations thereto. In some embodiments, the modified Cav-1 peptide comprises an additional 1-5 amino acids at either the N-terminus or C-terminus or both termini of any one of SEQ ID NOs: 2-111.

[0103] In some embodiments, the modified Cav-1 peptide comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 4-20. In some embodiments, the modified Cav-1 peptide comprises an amino acid sequence having at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 4-20. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of any one of SEQ ID NOs: 4-20 with one or more mutations thereto. For example, in some embodiments, the modified Cav-1 peptide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations thereto. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of any one of SEQ ID NOs: 4-20 with 1-5, 5-10, or 11-15, or more mutations thereto. In some embodiments, the modified Cav-1 peptide comprises an additional 1-5 amino acids at either the N-terminus or C-terminus or both termini of any one of SEQ ID NOs: 4-20.

[0104] In some embodiments, the modified Cav-1 peptide comprises or consists of the amino acid sequence of SEQ ID NO:2. In some embodiments, the modified Cav-1 peptide comprises an amino acid sequence having at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 95% sequence identity to SEQ ID NO:2. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of SEQ ID NO:2 with one or more mutations thereto. For example, in some embodiments, the modified Cav-1 peptide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations relative to SEQ ID NO:2. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of SEQ ID NO:2 with 1-5, 5-10, or 11-15, or more mutations thereto. In some embodiments, the modified Cav-1 peptide comprises an additional 1-5 amino acids at either or both the N-terminus or C-terminus of SEQ ID NO:2. In some embodiments, the modified Cav-1 peptide of SEQ ID NO:2 comprises an N-terminal and / or a C-terminal modification. In some embodiments, the N-terminal modification is an acylation. In some embodiments, the C-terminal modification is an amidation.

[0105] In some embodiments, the modified Cav-1 peptide comprises or consists of the amino acid sequence of SEQ ID NO:3. In some embodiments, the modified Cav-1 peptide comprises an amino acid sequence having at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 85% sequence identity to SEQ ID NO:3. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of SEQ ID NO:3 with one or more mutations thereto. For example, in some embodiments, the modified Cav-1 peptide comprises 1, 2, 3, 4, or 5 mutations relative to SEQ ID NO:3. In some embodiments, the modified Cav-1 peptide comprises an additional 1-5 amino acids at either or both the N-terminus or C-terminus of SEQ ID NO:3. In some embodiments, the modified Cav-1 peptide of SEQ ID NO:3 comprises an N-terminal modification and / or a C-terminal modification. In some embodiments, the N-terminal modification is an acylation. In some embodiments, the C-terminal modification is an amidation.

[0106] In some embodiments, the modified Cav-1 peptide comprises or consists of the amino acid sequence of SEQ ID NO:4. In some embodiments, the modified Cav-1 peptide comprises an amino acid sequence having at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, or at least 92% sequence identity to SEQ ID NO:4. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of SEQ ID NO:4 with one or more mutations thereto. For example, in some embodiments, the modified Cav-1 peptide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations relative to SEQ ID NO:4. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of SEQ ID NO:4 with 1-5, 5-10, or more mutations thereto. In some embodiments, the modified Cav-1 peptide comprises an additional 1-5 amino acids at either or both the N-terminus or C-terminus of SEQ ID NO:4. In some embodiments, the modified Cav-1 peptide of SEQ ID NO:4 comprises an N-terminal modification and / or a C-terminal modification. In some embodiments, the N-terminal modification is acylation. In some embodiments, the C-terminal modification is amidation. In some embodiments, the modified Cav-1 peptide comprises or consists of the amino acid sequence of SEQ ID NO:5.

[0107] In some embodiments, the modified Cav-1 peptide comprises or consists of the amino acid sequence of SEQ ID NO:6. In some embodiments, the modified Cav-1 peptide comprises an amino acid sequence having at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 95% sequence identity to SEQ ID NO:6. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of SEQ ID NO:6 with one or more mutations thereto. For example, in some embodiments, the modified Cav-1 peptide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations relative to SEQ ID NO:6. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of SEQ ID NO:6 with 1-5, 5-10, or 11-15, or more mutations thereto. In some embodiments, the modified Cav-1 peptide comprises an additional 1-5 amino acids at either or both the N-terminus or C-terminus of SEQ ID NO:6. In some embodiments, the modified Cav-1 peptide of SEQ ID NO:6 comprises an N-terminal modification and / or a C-terminal modification. In some embodiments, the N-terminal modification is an acylation. In some embodiments, the C-terminal modification is an amidation. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of SEQ ID NO:7. In some embodiments, the modified Cav-1 peptide comprises or consists of the amino acid sequence of SEQ ID NO:8.

[0108] In some embodiments, the modified Cav-1 peptide comprises or consists of the amino acid sequence of SEQ ID NO:9. In some embodiments, the modified Cav-1 peptide comprises an amino acid sequence having at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, or at least 92% sequence identity to SEQ ID NO:9. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of SEQ ID NO:9 with one or more mutations thereto. For example, in some embodiments, the modified Cav-1 peptide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations relative to SEQ ID NO:9. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of SEQ ID NO:9 with 1-5, 5-10, or more mutations thereto. In some embodiments, the modified Cav-1 peptide comprises an additional 1-5 amino acids at either or both the N-terminus or C-terminus of SEQ ID NO:9. In some embodiments, the modified Cav-1 peptide of SEQ ID NO:9 comprises an N-terminal modification and / or a C-terminal modification. In some embodiments, the N-terminal modification is acylation. In some embodiments, the C-terminal modification is amidation. In some embodiments, the modified Cav-1 peptide comprises or consists of the amino acid sequence of SEQ ID NO:10.

[0109] In some embodiments, the modified Cav-1 peptide comprises one, two, three, four or more amino acid substitutions, deletions or insertions relative to the sequence of SEQ ID NO:1, e.g., to yield a polypeptide of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 residues.

[0110] In some embodiments, the modified Cav-1 peptide has an amino acid sequence of FTTFTVT (SEQ ID NO: 3) and 1-5 additional amino acids on the N-terminus and / or C-terminus. In some embodiments, the modified Cav-1 peptide has an amino acid sequence of FTTFTVT (SEQ ID NO: 3) and, optionally, 1-5 additional amino acids on the N-terminus and / or C-terminus.

[0111] In some embodiments, the modified Cav-1 peptides provided in the present disclosure exhibit similar or identical biological activity as the native Cav-1 polypeptide in in vitro or in vivo assays. In some embodiments, the modified Cav-1 peptides exhibit at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 99%, and any range derivable therein, such as about 70% to about 80%, more preferably about 81% to about 90%, or even more preferably about 91% to about 99%, of the activity of the native Cav-1 polypeptide. In some embodiments, the modified Cav-1 peptides have 100% or even higher activity than the native Cav-1 polypeptide. Assays for testing biological activity, e.g., antifibrotic activity, the ability to affect the expression of uPA, uPAR, and PAI-1 mRNA, or the ability to inhibit fibroblast proliferation, are well known in the art.

[0112] In some embodiments, the modified Cav-1 peptides of the present disclosure are fragments, derivatives, or variants of natural Cav-1 polypeptides. The peptides may be synthetic, recombinant, or chemically modified peptides isolated or produced using methods well known in the art. Modifications can be made to the N-terminus, C-terminus, or internal amino acids. The peptides may contain conservative or non-conservative amino acid changes, as described below. Polynucleotide changes may result in amino acid substitutions, additions, deletions, fusions, and truncations in the Cav-1 polypeptide encoded by the reference sequence. The peptides may also contain insertions, deletions, or substitutions of amino acids (and other molecules) that do not normally occur in the peptide sequence on which the modified variant is based, such as, but not limited to, insertions of L-amino acids, or insertions of non-standard amino acids such as ornithine that do not normally occur in human proteins.

[0113] A. Substitution In some embodiments, the modified Cav-1 peptide comprises one or more conservative amino acid substitutions. Conservative amino acid substitutions result from the replacement of one amino acid with another amino acid with similar structural and / or chemical properties, such as the replacement of leucine with isoleucine or valine, the replacement of aspartic acid with glutamic acid, or the replacement of threonine with serine. Thus, conservative substitutions of a particular amino acid sequence refer to the replacement of an amino acid that is not important for the activity of a polypeptide, or the replacement of an important amino acid with another amino acid with similar properties (e.g., acidic, basic, positive or negative charge, polar or non-polar, etc.) that do not reduce the activity of the peptide. Conservative substitution tables providing functionally similar amino acids are well known in the art. For example, each of the following six groups contains amino acids that are conservative substitutions for one another: 1) alanine (A), serine (S), threonine (T), 2) aspartic acid (D), glutamic acid (E), 3) asparagine (N), glutamine (Q), 4) arginine (R), lysine (K), 5) isoleucine (I), leucine (L), methionine (M), valine (V), and 6) phenylalanine (F), tyrosine (Y), tryptophan (W). In some embodiments, individual substitutions, deletions, or additions that alter, add, or delete a single amino acid or a small percentage of amino acids may also be considered conservative substitutions if the change does not reduce the activity of the peptide. Insertions or deletions typically range from about 1 to 6 amino acids.

[0114] In some embodiments, the amino acid that will replace an existing amino acid can be selected based on the position of the existing amino acid, for example, the existing amino acid is on the outer surface of a peptide or polypeptide compared to the amino acid that is exposed to solvent or is not exposed to solvent and is located inside.The selection of such conservative amino acid substitutions is well known in the art, for example, as described in Dordo et al, J.Mol Biol, 1999, 217, 721-739 and Taylor et al, J.Theor.Biol.119(1986);205-218 and S.French and B.Robson, J.Mol.Evol.19(1983)171. For example, the following conservative amino acid substitutions suitable for the exterior surface of a peptide or polypeptide can be used: substitution of Y with F, substitution of T with S or K, substitution of P with A, substitution of E with D or Q, substitution of N with D or G, substitution of R with K, substitution of G with N or A, substitution of T with S or K, substitution of D with N or E, substitution of I with L or V, substitution of F with Y, substitution of S with T or A, substitution of G with N or A, substitution of K with R, substitution of A with S, K or P.

[0115] In some embodiments, conservative amino acid substitutions can be selected that are suitable for the interior of a peptide or polypeptide, where, for example, the amino acid is not exposed to solvent. For example, the following conservative amino acid substitutions can be used for the interior of a peptide or polypeptide: Y is replaced by F, T is replaced by A or S, I is replaced by L or V, W is replaced by Y, M is replaced by L, N is replaced by D, G is replaced by A, T is replaced by A or S, D is replaced by N, I is replaced by L or V, F is replaced by Y or L, S is replaced by A or T, and A is replaced by S, G, T or V. In some embodiments, non-conservative amino acid substitutions are also encompassed within the term variant.

[0116] In some embodiments, amino acid substitutions can be made in a polypeptide at one or more positions where the substitutions are for amino acids with similar hydrophilicity. The importance of hydrophobic amino acid index in conferring interactive biological functions to proteins is generally understood in the art. It is well-established that the relative hydrophobicity properties of amino acids contribute to the secondary structure of the resulting protein, which in turn determines the interaction of the protein with other molecules, such as enzymes, substrates, receptors, DNA, antibodies, antigens, etc. Thus, such conservative substitutions can be made in polypeptides and are likely to have only minor effects on their activity. As detailed in U.S. Pat. No. 4,554,101, the following hydrophilicity values ​​have been assigned to amino acid residues: arginine (+3.0), lysine (+3.0), aspartic acid (+3.0±1), glutamic acid (+3.0±1), serine (+0.3), asparagine (+0.2), glutamine (+0.2), glycine (0), threonine (−0.4), proline (−0.5±1), alanine (0.5), histidine −0.5), cysteine ​​(−1.0), methionine (−1.3), valine (−1.5), leucine (−1.8), isoleucine (−1.8), tyrosine (−2.3), phenylalanine (−2.5), tryptophan (−3.4), and these values ​​can be used as a guide. Thus, any of the modified Cav-1 peptides described herein may be modified by replacing an amino acid with a different but homologous amino acid with a similar hydrophilicity value. Amino acids with hydrophilicity within + / - 1.0 points or within + / - 0.5 points are considered to be homologous. In some embodiments, the modified Cav-1 peptide comprises a substitution of an amino acid with a hydrophilicity value within ±2. In some embodiments, the modified Cav-1 peptide comprises a substitution of an amino acid with a hydrophilicity value within ±1. In some embodiments, the modified Cav-1 peptide comprises a substitution of an amino acid with a hydrophilicity value within ±0.5.

[0117] In some embodiments, the modified Cav-1 peptides include non-natural amino acids. In some embodiments, the modified Cav-1 peptides include a combination of natural and non-natural amino acids, or only non-natural amino acids. The non-natural amino acids may include synthetic non-natural amino acids, substituted amino acids, or one or more D-amino acids (or other components of the composition, except for protease recognition sequences), as desired in certain situations. D-amino acid-containing peptides show increased stability in vitro or in vivo compared to L-amino acid-containing forms. Thus, constructing peptides incorporating D-amino acids may be particularly useful when greater in vivo or intracellular stability is desired or required. More specifically, D-peptides are resistant to endogenous peptidases and proteases, thereby resulting in better oral, transepithelial, and transdermal delivery of linked agents and conjugates, improved bioavailability of membrane-resident complexes, and extended intravascular and interstitial longevity when such properties are desired. Furthermore, D-peptides are poor at inducing humoral immune responses in the whole organism because they cannot be efficiently processed for major histocompatibility complex class II restricted presentation to T helper cells.

[0118] In addition to the 20 "standard" L-amino acids, D-amino acids that are clearly defined in the art, or non-standard, modified, or unusual amino acids are also contemplated for use in the present disclosure. Phosphorylated amino acids (Ser, Thr, Tyr), glycosylated amino acids (Ser, Thr, Asn), β-amino acids, GABA, ω-amino acids are further contemplated for use in the present disclosure. These include, for example, β-alanine (β-Ala) and other ω-amino acids, such as 3-aminopropionic acid, 2,3-diaminopropionic acid (Dpr), 4-aminobutyric acid, etc.; α-aminoisobutyric acid (Aib); ε-aminohexanoic acid (Aha); δ-aminovaleric acid (Ava); N-methylglycine or sarcosine (MeGly); ornithine (Orn); citrulline (Cit); t-butylalanine (t-BuA); t-butylglycine (t-BuG); N-methylisoleucine (MeIle); phenylglycine (P hg); norleucine (Nle); 4-chlorophenylalanine (Phe(4-Cl)); 2-fluorophenylalanine (Phe(2-F)); 3-fluorophenylalanine (Phe(3-F)); 4-fluorophenylalanine (Phe(4-F)); penicillamine (Pen); 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (Tic); homoarginine (hArg); N-acetyl-lysine (AcLys); 2,4-diaminobutyric acid (Dbu, Dab); p-aminophenylalanine (Phe(pNH 2 )); N-methylvaline (MeVal); homocysteine ​​(hCys); homophenylalanine (hPhe); and homoserine (hSer); hydroxyproline (Hyp); homoproline (hPro); N-methylated amino acids; and peptoids (N-substituted glycines). B. Derivatives

[0119] In some embodiments, the modified Cav-1 peptide is a derivative of a native Cav-1 polypeptide. The term "derivative" as used herein refers to a Cav-1 peptide that has been chemically modified by using techniques including, but not limited to, acetylation, ubiquitination, labeling, pegylation (derivatization with polyethylene glycol), lipidation, glycosylation, amidation, cyclization, or addition of other molecules. In some embodiments, the peptide is provided in a cyclic form, e.g., as a cyclic peptide, or as a lactam. Alternatively, or in addition, in some embodiments, the peptide is provided as a branched peptide. A molecule is also a "derivative" of another molecule when it contains additional chemical moieties that are not normally part of that molecule. Such moieties can change the pH or improve the solubility, absorption, biological half-life, etc. of the molecule. Alternatively, the moieties can reduce the toxicity of the molecule, eliminate or attenuate any undesirable side effects of the molecule, etc. Moieties capable of mediating such effects are disclosed in Remington's Pharmaceutical Sciences, 18th edition, A. R. Gennaro, Ed., Mack Publ., Easton, Pa. (1990), which is incorporated herein by reference in its entirety.

[0120] The term "functional" when used in conjunction with "derivative" or "variant" refers to a modified Cav-1 peptide that has a biological activity (either functional or structural) substantially similar to that of the entity or molecule that is its functional derivative or variant (e.g., a native Cav-1 polypeptide). The term functional derivative is intended to include fragments, analogs, or chemical derivatives of a molecule.

[0121] In some embodiments, the modified Cav-1 peptide includes co-translational and post-translational (e.g., C-terminal peptide cleavage) modifications such as, for example, disulfide bond formation, glycosylation, acetylation, phosphorylation, proteolytic cleavage (e.g., cleavage by furin or metalloproteases), to the extent that such modifications do not affect the function of the modified Cav-1 peptide.

[0122] In some embodiments, the modified Cav-1 peptide is a "retro-inverso peptide". A "retro-inverso peptide" refers to a peptide that has a reversed peptide bond orientation at at least one position, i.e., the amino and carboxy termini relative to the amino acid side chains. Thus, a retro-inverso analog has the peptide bond terminals reversed and reversed in orientation while maintaining the side chain topology largely as in the native peptide sequence. Retro-inverso peptides may contain L- or D-amino acids, or a mixture of L- and D-amino acids, with up to all amino acids being D-isomers. A partial retro-inverso peptide analog is a polypeptide in which only a portion of the sequence is reversed and replaced by enantiomeric amino acid residues. The retro-inverso portion of such an analog has reversed amino and carboxy termini, such that the amino acid residues adjacent to the retro-inverso portion are replaced by side chain analogs a substituted geminal-diaminomethane and malonate, respectively. Retro-inverso forms of cell membrane permeable peptides have been found to act as efficiently as natural forms in translocating across membranes.The synthesis of retro-inverso peptide analogs is described in Bonelli, F. et al., Int J Pept Protein Res.24(6):553-6(1984); Verdini, A and Viscomi, GC, J.Chem.Soc.Perkin Trans.1:697-701(1985) and U.S. Patent No. 6,261,569, the entire contents of which are incorporated herein by reference.

[0123] C. Terminal modifications In some embodiments, the Cav-1 peptides of the present disclosure are modified at their amino or carboxy termini (if linear). Examples of amino terminal modifications include, for example, N-glycosylated, N-alkylated, N-acetylated, or N-acylated amino acids. Terminal modifications may include pegylation. One example of a carboxy terminal modification is a C-terminal amidated amino acid. In some embodiments, the peptides are cross-linked or have cross-linking sites (e.g., modified Cav-1 peptides have cysteinyl residues and thus form cross-linked dimers in vitro or in vivo). In some embodiments, one or more peptidyl bonds are replaced by non-peptidyl bonds, the N-terminus or C-terminus are replaced, individual amino acid moieties are modified by treatment with agents capable of reacting with selected side chains or terminal residues, etc. Either the C-terminus or N-terminus of the amino acid sequence, or both, may be linked to a carboxylic acid or amine functional group, respectively. In some embodiments, modified Cav-1 peptides include N-terminal modifications. In some embodiments, the modified Cav-1 peptide comprises a C-terminal modification. In some embodiments, the modified Cav-1 peptide comprises an N-terminal and a C-terminal modification.

[0124] Non-limiting illustrative examples of N-terminal protecting groups include acyl groups (-CO-R1) and alkoxycarbonyl or aryloxycarbonyl groups (-CO-O-R1), where R1 is an aliphatic, substituted aliphatic, benzyl, substituted benzyl, aromatic, or substituted aromatic group. Specific examples of acyl groups include, but are not limited to, acetyl, (ethyl)-CO-, n-propyl-CO-, iso-propyl-CO-, n-butyl-CO-, sec-butyl-CO-, t-butyl-CO-, hexyl, lauroyl, palmitoyl, myristoyl, stearyl, oleoyl, phenyl-CO-, substituted phenyl-CO-, benzyl-CO-, and (substituted benzyl)-CO-. Examples of alkoxycarbonyl and aryloxycarbonyl groups include, but are not limited to, CH 3-O-CO-, (ethyl)-O-CO-, n-propyl-O-CO-, isopropyl-O-CO-, n-butyl-O-CO-, sec-butyl-O-CO-, t-butyl-O-CO-, phenyl-O-CO-, (substituted phenyl)-O-CO-, benzyl-O-CO-, and (substituted benzyl)-O-CO-. To facilitate N-acylation, one to four glycine residues may be present at the N-terminus of the molecule.

[0125] Carboxy terminal modifications include acylation with carboxylic acids: formic acid, acetic acid, propionic acid, fatty acids (myristic acid, palmitic acid, stearic acid), succinic acid, and benzoic acid; carbonylation (such as benzyloxycarbonylation (Cbz)); acetylation; and biotinylation. Amino terminal modifications include, but are not limited to, (i) acylation with carboxylic acids: formic acid, acetic acid, propionic acid, fatty acids (such as myristic acid, palmitic acid, stearic acid), succinic acid, and benzoic acid; (ii) carbonylation (such as benzyloxycarbonylation (Cbz)); (iii) biotinylation; (iv) amidation; and (v) fluorescein (FITC, FAM, etc.), 7-hydroxy-4-methylcoumarin-3-acetic acid, 7-hydroxy ... These include phosphorus-3-acetic acid, 7-methoxycoumarin-3-acetic acid, and other coumarins; rhodamine (5-carboxyrhodamine 110 or 6G, 5(6)-TAMRA, ROX); attachment of dyes such as N-[4-(4-dimethylamino)phenylazo]benzoic acid (Dabcyl), 2,4-dinitrobenzene (Dnp), 5-dimethylaminonaphthalene-1-sulfonic acid (Dansyl), and other dyes; and (vi) PEGylation.

[0126] The carboxyl group at the C-terminus of a peptide may be, for example, converted to an amide (i.e., the hydroxyl group at the C-terminus is -NH 2, -NHR2, and -NR2R3) or esters (i.e., the C-terminal hydroxyl group is replaced by -OR2). R2 and R3 are optionally independently aliphatic, substituted aliphatic, benzyl, substituted benzyl, aryl, or substituted aryl groups. Additionally, together with the nitrogen atom, R2 and R3 can optionally form a C4-C8 heterocycle having about 0-2 additional heteroatoms such as nitrogen, oxygen, or sulfur. Non-limiting examples of heterocycles include, but are not limited to, piperidinyl, pyrrolidinyl, morpholinyl, thiomorpholinyl, or piperazinyl. Examples of C-terminal protecting groups include, but are not limited to, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NH(ethyl), -N(ethyl) 2 , -N(methyl)(ethyl), -NH(benzyl), -N(C 1 -C 4 alkyl)(benzyl), -NH(phenyl), -N(C 1 -C 4 Alkyl)(phenyl), -OCH 3 , -O-(ethyl), -O-(n-propyl), -O-(n-butyl), -O-(isopropyl), -O-(sec-butyl), -O-(t-butyl), -O-benzyl, and -O-phenyl.

[0127] D. Side chain modifications In some embodiments, the modified Cav-1 peptide of the present disclosure comprises modified amino acid side chains. Non-limiting examples of modifications include carboxymethylation, acylation, phosphorylation, glycosylation, or fatty acylation. An ether bond can optionally be used to link a serine or threonine hydroxyl to a hydroxyl of a sugar. An amide bond can optionally be used to link a carboxyl group of glutamic acid or aspartic acid to an amino group on a sugar (Gang and Jeanloz, Advances in Carbohydrate Chemistry and Biochemistry, Vol. 43, Academic Press (1985); Kunz, Ang. Chem. Int. Ed. English 26: 294-308 (1987)). Acetyl and ketal bonds can also optionally be formed between amino acids and carbohydrates. Fatty acyl derivatives can optionally be generated, for example, by acylation of free amino groups (e.g., lysine) (Toth et al., Peptides: Chemistry, Structure and Biology, Rivier and Marshal, eds., ESCOM Publ., Leiden, 1078-1079 (1990)).

[0128] As used herein, the term "chemically modified", when referring to modified Cav-1 peptides of the present disclosure, refers to a peptide in which at least one of its amino acid residues is modified by natural processes such as processing or other post-translational modifications, or by chemical modification techniques well known in the art. Examples of numerous known modifications typically include, but are not limited to, acetylation, acylation, amidation, ADP-ribosylation, glycosylation, GPI anchor formation, covalent attachment of lipids or lipid derivatives, methylation, myristylation, pegylation, prenylation, phosphorylation, ubiquitination, or any similar process.

[0129] Other types of modifications optionally include the addition of cycloalkane moieties to biomolecules such as proteins, as described in PCT application WO2006 / 050262, which is incorporated herein by reference in its entirety. These moieties are designed for use with biomolecules and can optionally be used to confer various properties to the proteins.

[0130] In addition, any point on the protein can be optionally modified. For example, pegylation of glycosylation moieties on the protein can be optionally performed as described in PCT application WO2006 / 050247, the entirety of which is incorporated herein by reference. One or more polyethylene glycol (PEG) groups can be optionally added to O-linked and / or N-linked glycosylation. The PEG groups can be optionally branched or linear. Optionally, any type of water-soluble polymer can be added to the glycosylation site on the protein via a glycosyl linker.

[0131] Covalent modifications of the modified Cav-1 peptides of the present disclosure are included within the scope of the present invention. Other types of covalent modifications of peptides are introduced into the molecule by reacting targeted amino acid residues with organic derivatizing agents that can react with selected side chains or N- or C-terminal residues.

[0132] Cysteinyl residues most commonly are reacted with α-haloacetates (and corresponding amines), such as chloroacetic acid or chloroacetamide, to give carboxymethyl or carboxyamidomethyl derivatives. Cysteinyl residues are also derivatized by reaction with bromotrifluoroacetone, α-bromo-β-(5-imidozoyl)propionic acid, chloroacetylphosphate, N-alkylmaleimides, 3-nitro-2-pyridyl disulfide, 2-pyridyl disulfide methyl, p-chloromercuribenzoate, 2-chloromercuri-4-nitrophenol, or chloro-7-nitrobenzo-2-oxa-1,3-diazole.

[0133] Histidyl residues are derivatized by reaction with diethylpyrocarbonate at pH 5.5-7.0 because that agent is relatively specific for the histidyl side chain. Para-bromophenacyl bromide is also useful, and the reaction is preferably performed in 0.1 M sodium cacodylate at pH 6.0.

[0134] Lysinyl and amino-terminal residues react with succinic or other carboxylic acid anhydrides. Derivatization with these agents has the effect of reversing the charge of the lysinyl residues. Other suitable reagents for derivatizing α-amino-containing residues include transaminase-catalyzed reactions with methyl picolinimidate, pyridoxal phosphate, pyridoxal, chloroborohydride, trinitrobenzenesulfonic acid, O-methylisourea, 2,4-pentanedione, and glyoxylic acid.

[0135] Arginyl residues are modified by reaction with one or several conventional reagents, including phenylglyoxal, 2,3-butanedione, 1,2-cyclohexanedione, and ninhydrin.

[0136] Derivatization of arginine residues requires that the reaction be performed under alkaline conditions due to the high pKa of the guanidine functional group. Moreover, these reagents can react with lysine as well as arginine epsilon-amino groups.

[0137] The specific modification of tyrosyl residues can be made, with particular interest in introducing spectral labels into tyrosyl residues by reaction with aromatic diazonium compounds or tetranitromethane. Most commonly, N-acetylimidazole and tetranitromethane are used to form O-acetyltyrosyl species and 3-nitro derivatives, respectively. Tyrosyl residues are iodized with 125I or 131I to prepare labeled peptides for use in radioimmunoassay.

[0138] Carboxyl side groups (aspartyl or glutamyl) are selectively modified by reaction with carbodiimides (RN=C=N-R'), where R and R' are different alkyl groups, such as 1-cyclohexyl-3-(2-morpholinyl-4-ethyl)carbodiimide or 1-ethyl-3-(4-azonia-4,4-dimethylpentyl)carbodiimide. Furthermore, aspartyl and glutamyl residues are converted to asparaginyl and glutaminyl residues by reaction with ammonium ions.

[0139] Derivatization with bifunctional agents is useful for crosslinking to water-insoluble support matrices or surfaces for use in methods of purifying anti-CHF antibodies, and vice versa. Commonly used crosslinking agents include, for example, 1,1-bis(diazoacetyl)-2-phenylethane, glutaraldehyde, N-hydroxysuccinimide esters, such as esters with 4-azidosalicylic acid, homobifunctional imidoesters including disuccinimidyl esters, such as 3,3'-dithiobis(succinimidylpropionic acid), and bifunctional maleimides, such as bis-N-maleimido-1,8-octane. Derivatizing agents such as methyl-3-[(p-azidophenyl)dithio]propioimidate produce photoactivatable intermediates that can form crosslinks in the presence of light. Alternatively, reactive water-insoluble substrates such as cyanogen bromide-activated carbohydrates and reactive substrates described in U.S. Patent Nos. 3,969,287, 3,691,016, 4,195,128, 4,247,642, 4,229,537, and 4,330,440 are employed for protein immobilization.

[0140] Glutaminyl and asparaginyl residues are frequently deamidated to the corresponding glutamyl and aspartyl residues, respectively. These residues are deamidated under neutral or basic conditions. The deamidated forms of these residues are within the scope of the invention.

[0141] Other modifications include hydroxylation of proline and lysine, phosphorylation of the hydroxyl groups of seryl or threonyl residues, methylation of the α-amino groups of lysine, arginine, and histidine side chains (TECreighton, Proteins: Structure and Molecular Properties, W.H. Freeman & Co., San Francisco, pp. 79-86

[1983] ), acetylation of the N-terminal amine, and amidation of any C-terminal carboxyl group.

[0142] E. Capping In some embodiments, the modified Cav-1 peptide is capped at its N-terminus and / or C-terminus. In some embodiments, the modified Cav-1 peptide is capped at its N-terminus and / or C-terminus with an acyl (abbreviated "Ac") group and / or an amide (abbreviated "Am") group, respectively, e.g., acetyl (CH) at the N-terminus. 3 CO-) group and the amide (-NH 2 In some embodiments, the modified Cav-1 peptide is capped at its N-terminus with an acyl group, for example, an acetyl (CH) group at the N-terminus. 3 In some embodiments, the modified Cav-1 peptide is capped with an amide group at its C-terminus, e.g., an amide (-NH 2 ) is capped.

[0143] In some embodiments, the modified Cav-1 peptide is capped at its N-terminus. A wide variety of N-terminal capping functional groups, preferably linked to the terminal amino group, are envisaged, for example: Formyl, Alkanoyl having 1 to 10 carbon atoms, such as acetyl, propionyl, butyryl, etc. Alkenoyl having 1 to 10 carbon atoms, such as hex-3-enoyl; Alkynoyl having 1 to 10 carbon atoms, such as hex-5-ynoyl; aroyl, such as benzoyl or 1-naphthoyl; heteroaroyl, such as 3-pyrroyl or 4-quinoloyl; Alkylsulfonyl, such as methanesulfonyl, Arylsulfonyl, such as benzenesulfonyl or sulfanilyl; Heteroarylsulfonyl, such as pyridine-4-sulfonyl; Substituted alkanoyl having 1 to 10 carbon atoms, such as 4-aminobutyryl; Substituted alkenoyl having 1 to 10 carbon atoms, such as 6-hydroxy-hex-3-enoyl; Substituted alkynoyl having 1 to 10 carbon atoms, such as 3-hydroxy-hex-5-ynoyl; Substituted aroyl, such as 4-chlorobenzoyl or 8-hydroxy-naphth-2-yl; Substituted heteroaroyl, such as 2,4-dioxo-1,2,3,4-tetrahydro-3-methyl-quinazolin-6-oyl; Substituted alkylsulfonyl, such as 2-aminoethanesulfonyl; Substituted arylsulfonyl, such as 5-dimethylamino-1-naphthalenesulfonyl; substituted heteroarylsulfonyl, such as 1-methoxy-6-isoquinolinesulfonyl; Carbamoyl or thiocarbamoyl, substituted carbamoyl (R'-NH-CO) or substituted thiocarbamoyl (R'-NH-CS), where R' is alkyl, alkenyl, alkynyl, aryl, heteroaryl, substituted alkyl, substituted alkenyl, substituted alkynyl, substituted aryl, or substituted heteroaryl; Substituted carbamoyl (R'-NH-CO) and substituted thiocarbamoyl (R'-NH-CS), where R' is alkanoyl, alkenoyl, alkynoyl, aroyl, heteroaroyl, substituted alkanoyl, substituted alkenoyl, substituted alkynoyl, substituted aroyl, or substituted heteroaroyl, all as defined above.

[0144] In some embodiments, the modified Cav-1 peptide is capped at its C-terminus. The C-terminal capping functional group can be either an amide bond or an ester bond with the terminal carboxyl. The capping functional group that provides an amide bond is NR 1 R 2 where R 1 and R 2 may be independently selected from the following group: hydrogen, Preferably, alkyl having 1 to 10 carbon atoms, such as methyl, ethyl, isopropyl, etc. Preferably, alkenyl having 1 to 10 carbon atoms, such as prop-2-enyl, Preferably, alkynyl having 1 to 10 carbon atoms, such as prop-2-ynyl, Substituted alkyls having 1 to 10 carbon atoms, such as hydroxyalkyls, alkoxyalkyls, mercaptoalkyls, alkylthioalkyls, halogenoalkyls, cyanoalkyls, aminoalkyls, alkylaminoalkyls, dialkylaminoalkyls, alkanoylalkyls, carboxyalkyls, carbamoylalkyls, etc. Substituted alkenyls having 1 to 10 carbon atoms, such as hydroxyalkenyls, alkoxyalkenyls, mercaptoalkenyls, alkylthioalkenyls, halogenoalkenyls, cyanoalkenyls, aminoalkenyls, alkylaminoalkenyls, dialkylaminoalkenyls, alkanoylalkenyls, carboxyalkenyls, carbamoylalkenyls, etc. Substituted alkynyls having 1 to 10 carbon atoms, such as hydroxyalkynyl, alkoxyalkynyl, mercaptoalkynyl, alkylthioalkynyl, halogenoalkynyl, cyanoalkynyl, aminoalkynyl, alkylaminoalkynyl, dialkylaminoalkynyl, alkanoylalkynyl, carboxyalkynyl, carbamoylalkynyl, etc. aroylalkyl having up to 10 carbon atoms, such as phenacyl or 2-benzoylethyl; aryl, such as phenyl or 1-naphthyl; Heteroaryl, such as 4-quinolyl, Alkanoyl having 1 to 10 carbon atoms, such as acetyl or butyryl; Aroyl, such as benzoyl, Heteroaroyl, such as 3-quinoloyl, OR' or NR'R'', where R' and R'' are independently hydrogen, alkyl, aryl, heteroaryl, acyl, aroyl, sulfonyl, sulfinyl, or SO 2 -R''' or SO-R''', where R'' is a substituted or unsubstituted alkyl, aryl, heteroaryl, alkenyl, or alkynyl.

[0145] Capping functionalities providing an ester linkage are designated OR, where R can be alkoxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, substituted alkoxy, substituted aryloxy, substituted heteroaryloxy, substituted aralkyloxy, or substituted heteroaralkyloxy.

[0146] In some embodiments, the N-terminal or C-terminal capping functional groups, or both, are structured such that the capping molecule undergoes spontaneous or enzymatic conversion in the body to release the active drug, thereby functioning as a prodrug (a pharmacologically inactive derivative of a parent drug molecule) with improved delivery properties over the parent drug molecule (Bundgaard H, Ed: Design of Prodrugs, Elsevier, Amsterdam, 1985).

[0147] Careful selection of the capping group can allow for the addition of other activities to the peptide, for example, the presence of a sulfhydryl group linked to the N- or C-terminal cap allows for the conjugation of the derivatized peptide to other molecules.

[0148] F. Multimer formation Embodiments of the present disclosure also include longer polypeptides constructed from repeating units of modified Cav-1 peptides. In some embodiments, the polypeptide multimers include different combinations of polypeptides. In some embodiments, the multimeric polypeptides are made by chemical synthesis or by recombinant DNA techniques as discussed herein. When produced by chemical synthesis, the oligomers, in some embodiments, preferably have 2-5 repeats of the core polypeptide sequence, and the total number of amino acids in the multimer should not exceed about 160 residues, preferably 100 residues or less (or their equivalents if linkers or spacers are included).

[0149] In some embodiments, the modified Cav-1 peptide is a multimer comprising at least two peptides of the present disclosure. In some embodiments, a first peptide of the at least two peptides is essentially identical to a second peptide of the at least two peptides. In some embodiments, a first peptide of the at least two peptides is not identical to a second peptide of the at least two peptides.

[0150] G. Peptide Mimetics In some embodiments, modified Cav-1 peptide is a peptidomimetic compound that mimics the biological effect of natural Cav-1 polypeptide.In some embodiments, peptidomimetics are non-natural peptides or non-peptide agents that reproduce the spatial properties of the binding elements of natural Cav-1 polypeptide so as to have the binding activity and biological activity of natural Cav-1 polypeptide.Like natural Cav-1 polypeptide or polypeptide multimers, peptidomimetics have a binding surface that interacts with any ligand that natural Cav-1 polypeptide binds, and a non-binding surface.

[0151] In some embodiments, the present disclosure also includes modified Cav-1 peptides that retain partial peptidic properties. For example, any proteolytically unstable bonds in the modified Cav-1 peptides can be selectively replaced by non-peptide elements such as isosteres (N-methylated, D-amino acids) or reduced peptide bonds, while the remainder of the molecule retains its peptidic properties.

[0152] Peptidomimetic compounds, either agonists, substrates or inhibitors, have been described for several biologically active peptides / polypeptides, such as opioid peptides, VIP, thrombin, HIV protease, etc. Methods for designing and preparing peptidomimetic compounds are known in the art (Hruby, VJ, Biopolymers 33:1073-1082(1993); Wiley, RA et al., Med.Res.Rev.13:327-384(1993); Moore et al., Adv.in Pharmacol 33:91-141(1995); Giannis et al., Adv.in Drug Res.29:1-78(1997)). Certain mimetics that mimic secondary structures are described in Johnson et al., In:Biotechnology and Pharmacy, Pezzuto et al., Chapman and Hall(Eds.), NY, 1993. These methods are used to generate peptide mimetics that have at least the binding ability and specificity of the native Cav-1 polypeptide, and preferably also have biological activity. Knowledge of peptide chemistry and general organic chemistry available to those skilled in the art is sufficient to design and synthesize such compounds, given the present disclosure.

[0153] For example, such peptidomimetics can be identified by examining the three-dimensional structure of the polypeptide of the present invention, either free or bound in complex with a ligand (e.g., soluble uPAR or its fragment). Alternatively, the structure of the polypeptide of the present invention bound to its ligand can be obtained by the technique of nuclear magnetic resonance spectroscopy. Further knowledge of the stereochemistry of the interaction of the peptide with its ligand or receptor allows such peptidomimetics to be rationally designed. The structure of the peptide or polypeptide of the present invention in the absence of ligand can also provide a scaffold for the design of mimetic molecules.

[0154] H. PEGylation In some embodiments, the modified Cav-1 peptide of the present disclosure is conjugated with a heterologous polypeptide segment or polymer, such as polyethylene glycol.In some embodiments, the modified Cav-1 peptide is linked to PEG to increase the hydrodynamic radius of the enzyme and thus increase serum persistence.In some embodiments, the modified Cav-1 peptide is conjugated with any targeting agent, such as a ligand that has the ability to specifically and stably bind to external receptors (see, for example, US Patent Application Publication No. 2009 / 0304666).

[0155] In some embodiments, the present disclosure provides methods and compositions related to PEGylation of Cav-1 peptides. PEGylation is a process of covalently attaching a poly(ethylene glycol) polymer chain to another molecule, usually a drug or a therapeutic protein. PEGylation is routinely achieved by incubating a reactive derivative of PEG with a target macromolecule. Covalent attachment of PEG to a drug or therapeutic protein can either "mask" the drug from the host's immune system (reducing immunogenicity and antigenicity) or increase the hydrodynamic size (size of the solution) of the drug, thereby extending its circulation time by reducing renal clearance. PEGylation can also provide water solubility to hydrophobic drugs and proteins.

[0156] The first step in PEGylation is the appropriate functionalization of the PEG polymer at one or both termini. PEG activated at each end with the same reactive moiety is known as "homobifunctional", whereas when the functional groups present are different, the PEG derivative is termed "heterobifunctional" or "heterofunctional". Chemically active or activated derivatives of PEG polymers are prepared to allow the attachment of PEG to the desired molecule.

[0157] The selection of suitable functional groups for PEG derivatives is based on the type of available reactive groups on the modified Cav-1 peptide to be attached to PEG. For proteins, typical reactive amino acids include lysine, cysteine, histidine, arginine, aspartic acid, glutamic acid, serine, threonine, and tyrosine. N-terminal amino groups and C-terminal carboxylic acid groups can also be used.

[0158] Techniques used to form first generation PEG derivatives generally react PEG polymers with groups that react with hydroxyl groups, typically anhydrides, acid chlorides, chloroformates, and carbonates. In second generation PEGylation chemistries, more efficient functional groups such as aldehydes, esters, and amides become available for conjugation.

[0159] As PEGylation applications become more advanced and sophisticated, there is an increasing need for heterobifunctional PEGs for conjugation. These heterobifunctional PEGs are very useful for linking two entities where a hydrophilic, flexible, and biocompatible spacer is required. The preferred terminal groups for heterobifunctional PEGs are maleimide, vinyl sulfone, pyridyl disulfide, amine, carboxylic acid, and N-hydroxysuccinimide (NHS) ester.

[0160] The most common modifiers, or linkers, are based on methoxy PEG (mPEG) molecules. Their activity depends on the addition of protein-modifying groups to the alcohol termini. In some embodiments, polyethylene glycol (PEG diol) is used as a precursor molecule. The diol is then modified at both ends to create heterodimeric or homodimeric PEG-linked molecules.

[0161] Proteins are generally PEGylated at nucleophilic sites such as unprotonated thiols (cysteinyl residues) or amino groups. Examples of cysteinyl-specific modification reagents include PEG maleimide, PEG iodoacetate, PEG thiol, and PEG vinylsulfone. All four are cysteinyl-specific under mild conditions and have a neutral to slightly alkaline pH, but each has some drawbacks. The thioether formed by maleimide may be somewhat unstable under alkaline conditions, so formulation options with this linker may be somewhat limited. The carbamothioate bond formed by iodoPEG is more stable, but free iodine may modify tyrosine residues under some conditions. PEG thiol forms disulfide bonds with protein thiols, but this bond may also be unstable under alkaline conditions. The reactivity of PEG-vinylsulfone is relatively slow compared to maleimide and iodoPEG, but the thioether bond formed is very stable. Its slower reaction rate may also make it easier to control the PEG-vinylsulfone reaction.

[0162] Site-specific PEGylation at natural cysteinyl residues is rarely performed because these residues are usually in the form of disulfide bonds or are required for biological activity. Alternatively, site-directed mutagenesis can be used to incorporate cysteinyl PEGylation sites for use with thiol-specific linkers. The cysteine ​​mutations must be designed to be accessible to the PEGylation reagent and biologically active after PEGylation.

[0163] Amine-specific modifiers include PEG NHS ester, PEG tresylate, PEG aldehyde, PEG isothiocyanate, and several others. All react under mild conditions and are very specific for amino groups. PEG NHS ester is probably one of the more reactive agents, but its high reactivity can make the PEGylation reaction difficult to control on a large scale. PEG aldehyde forms an imine with the amino group, which is then reduced to a secondary amine by sodium cyanoborohydride. Unlike sodium borohydride, sodium cyanoborohydride does not reduce disulfide bonds. However, this chemical is highly toxic and must be handled with care, especially at low pH where it becomes volatile.

[0164] Site-specific PEGylation can be a challenge due to the presence of multiple lysine residues on most proteins. Because these reagents react with unprotonated amino groups, it is possible to target PEGylation to amino groups with lower pK by performing the reaction at a lower pH. In general, the pK of an alpha-amino group is 1-2 pH units lower than the epsilon-amino group of a lysine residue. By PEGylating molecules at pH 7 or lower, high selectivity for the N-terminus can frequently be achieved. However, this is only feasible if the N-terminal portion of the protein is not required for biological activity. Furthermore, the pharmacokinetic benefits from PEGylation frequently outweigh any significant loss of in vitro bioactivity, resulting in products with much greater in vivo bioactivity independent of the PEGylation chemistry.

[0165] There are several parameters to consider when developing a PEGylation procedure. Fortunately, there are usually no more than four or five critical parameters. A "design of experiments" approach to optimizing PEGylation conditions can be very useful. For thiol-specific PEGylation reactions, parameters to consider include protein concentration, PEG to protein ratio (on a molar basis), temperature, pH, reaction time, and possibly the exclusion of oxygen. (Oxygen can contribute to intermolecular disulfide formation with proteins, thereby reducing the yield of PEGylated product.) The same factors (except oxygen) should be considered for amine-specific modifications, especially when targeting N-terminal amino groups, except pH can be even more important.

[0166] For both amine-specific and thiol-specific modifications, the reaction conditions can affect the stability of the protein. This can include temperature, protein concentration, and pH. Furthermore, the reactivity of the PEG linker should be known before initiating the PEGylation reaction. For example, if the PEGylation agent is only 70% active, the amount of PEG used should ensure that only active PEG molecules are considered in the stoichiometry of the protein-PEG reaction.

[0167] I. Fusion Proteins In some embodiments, the disclosure provides fusion proteins of modified Cav-1 peptides. For example, fusions may employ leader sequences from other species to allow recombinant expression of the protein in a heterologous host. The fusion protein may include a half-life extender. Another useful fusion includes the addition of a protein affinity tag, such as a serum albumin affinity tag or six histidine residues, or an immunoactive domain, such as an antibody epitope, preferably a cleavable epitope, to facilitate purification of the fusion protein. Non-limiting affinity tags include polyhistidine, chitin binding protein (CBP), maltose binding protein (MBP), and glutathione-S-transferase (GST). In some embodiments, the modified Cav-1 peptide includes a heterologous peptide or protein linked at the N-terminus and / or C-terminus. In some embodiments, the heterologous peptide or protein is a leader sequence, a half-life extender, a protein affinity tag, or an immunoactive domain.

[0168] In some embodiments, the modified Cav-1 peptide is linked to a peptide that increases in vivo half-life, such as an XTEN® polypeptide (Schellenberger et al., 2009), an IgG Fc domain, albumin, or an albumin-binding peptide.

[0169] Methods for generating fusion proteins are well known to those skilled in the art. Such proteins can be produced, for example, by de novo synthesis of a complete fusion protein, or by the addition of a DNA sequence encoding a heterologous domain followed by expression of the intact fusion protein.

[0170] The production of fusion proteins that restore the functional activity of the parent protein can be facilitated by connecting the genes with a bridging DNA segment that encodes a peptide linker that is spliced ​​between the tandemly connected polypeptides, the linker being of sufficient length to allow proper folding of the resulting fusion protein.

[0171] 1. Linker In some embodiments, the modified Cav-1 peptides are chemically conjugated using bifunctional cross-linking reagents or fused at the protein level with a peptide linker.

[0172] Bifunctional cross-linking reagents have been widely used for various purposes, including the preparation of affinity matrices, the modification and stabilization of various structures, the identification of ligand-receptor binding sites, and structural studies. In some embodiments, a peptide linker, such as a Gly-Ser linker, is used to link the modified Cav-1 peptide of the present disclosure.

[0173] Homobifunctional reagents carrying two identical functional groups have proven to be very efficient in cross-linking between identical and different macromolecules or subunits of macromolecules, as well as in linking polypeptide ligands to their specific binding sites. Heterobifunctional reagents contain two different functional groups. By exploiting the differential reactivity of the two different functional groups, cross-linking can be controlled selectively and sequentially. Bifunctional cross-linking reagents can be divided according to the specificity of their functional groups, e.g., amino, sulfhydryl, guanidine, indole, carboxyl specific groups. Of these, reagents directed to free amino groups are particularly gaining popularity due to their commercial availability, ease of synthesis, and application in mild reaction conditions.

[0174] Most heterobifunctional cross-linking reagents contain a primary amine-reactive group and a thiol-reactive group. In another example, heterobifunctional cross-linking reagents and methods of using the cross-linking reagents are described (U.S. Pat. No. 5,889,155, which is incorporated herein by reference in its entirety). The cross-linking reagents combine a nucleophilic hydrazide residue with an electrophilic maleimide residue, allowing, in one example, the attachment of an aldehyde to a free thiol. The cross-linking reagents can be modified to cross-link a variety of functional groups.

[0175] Furthermore, any other linking / coupling agent and / or mechanism known to one of skill in the art may be used to combine the modified Cav-1 peptides of the present disclosure, such as, for example, antibody-antigen interactions, avidin-biotin bonds, amide bonds, ester bonds, thioester bonds, ether bonds, thioether bonds, phosphoester bonds, phosphoramide bonds, anhydride bonds, disulfide bonds, ionic and hydrophobic interactions, bispecific antibodies and antibody fragments, or combinations thereof.

[0176] In some embodiments, modified Cav-1 peptide comprises a crosslinker that has reasonable stability in blood.Many types of disulfide bond-containing linkers are known that can be successfully employed to conjugate targeting agents and therapeutic / prophylactic agents.Containing sterically hindered disulfide bond linkers may prove to provide greater stability in vivo.Therefore, in some embodiments, modified Cav-1 peptide comprises a sterically hindered crosslinker.

[0177] In addition to hindered crosslinkers, non-hindered linkers can also be employed according to the present specification. In some embodiments, modified Cav-1 peptide comprises non-sterically hindered crosslinkers. Other useful crosslinkers that are not considered to contain or generate protected disulfides include SATA, SPDP, and 2-iminothiolane (Wawrzynczak and Thorpe, 1987). The use of such crosslinkers is well understood in the art.

[0178] In some embodiments, the modified Cav-1 peptide comprises a flexible linker.

[0179] Once chemically conjugated, the modified Cav-1 peptide will generally be purified to separate the conjugate from non-conjugated agents and from other contaminants. A number of purification techniques are available for use in providing the conjugate of sufficient purity to confer clinical utility.

[0180] Purification methods based on size separation, such as gel filtration, gel permeation, or high performance liquid chromatography, are the most commonly used. Other chromatographic techniques, such as Blue Sepharose separation, can also be used. Conventional methods for purifying fusion proteins from inclusion bodies, such as using weak detergents, such as sodium N-lauroyl sarcosine (SLS), can be useful.

[0181] 2. Cell membrane-permeable peptides and membrane-translocating peptides In some embodiments, the modified Cav-1 peptide comprises a cell-binding domain or a cell membrane-penetrating peptide (CPP). As used herein, the terms "cell membrane-penetrating peptide", "membrane translocation domain" and "protein transduction domain" are used interchangeably and refer to a segment of a polypeptide sequence that allows the polypeptide to cross a cell membrane (e.g., the plasma membrane in the case of eukaryotic cells). Examples of CPPs include, but are not limited to, segments derived from HIV-binding peptides, HIV-1 Tat (HIV), Tat-derived peptides, penetratin, VP22-derived or analog peptides, HSV Examples include VP22 (Herpes simplex), Protegrin I, MAP, KALA or protein transduction domain (PTD), PpT620, proline-rich peptide, arginine-rich peptide, lysine-rich peptide, MPG-peptide, Pep-1, L-oligomer, calcitonin peptide, antennapedia-derived peptide (specifically from Drosophila antennapedia), pAntp, T1 (TKIESLKEHG, SEQ ID NO: 115), T2 (TQIENLKEKG, SEQ ID NO: 116), 26 (AALEALAEALEALAEALEALAEAAAA, SEQ ID NO: 117), INF7 (GLFEAIEGFIENGWEGMIEGWYGCG, SEQ ID NO: 118) pIsl, FGF, lactoferrin, transportan, buforin-2, Bac715-24, SynB, SynB(1), pVEC, hCT-derived peptide, SAP, or histones.

[0182] CPPs typically have an amino acid composition that either contains a high relative abundance of positively charged amino acids such as lysine or arginine, or has a sequence that contains an alternating pattern of polar / charged and non-polar hydrophobic amino acids. These two types of structures are called polycationic or amphipathic, respectively. Typically, CPPs are peptides of 8-50 residues that have the ability to cross cell membranes and enter most cell types. Frankel and Pabo described the ability of the transactivating transcription activator from human immunodeficiency virus 1 (HIV-TAT) to penetrate into cells (Frankel, AD and CO Pabo, Cellular uptake of the tat protein from human immunodeficiency virus. Cell, 1988.55(6):p.1189-93). In 1991, the introduction of the Antennapedia homeobox peptide (DNA binding domain) derived from Drosophila melanogaster into neural cells was also described (Joliot, A., et al., Antennapedia homeobox peptide regulates neural morphogenesis. Proc Natl Acad Sci USA, 1991.88(5):p.1864-8). In 1994, the first 16mer peptide CPP, called penetratin (RQIKIWFQNRRMKWKK, SEQ ID NO: 113), derived from the third helix of the homeodomain of the Drosophila Antennapedia homeobox gene product, was characterized (Derossi, D., et al., The third helix of the Antennapedia homeodomain translocates through biological membranes. J Biol Chem, 1994. 269(14): p. 10444-50), followed in 1998 by the characterization of the minimal domain of TAT (e.g., GRKKRRQRRRPPQ, SEQ ID NO: 112) required for protein transduction (Vives, E., P. Brodin, and B.Lebleu, A truncated HIV-1 Tat protein basic domain rapidly translocates through the plasma membrane and accumulates in the cell nucleus. J Biol Chem, 1997.272(25):p.16010-7). Over the past two decades, dozens of peptides from different sources have been described, including viral proteins such as herpesvirus VP22 (Elliott, G. and P. O'Hare, Intercellular trafficking and protein delivery by a herpesvirus structural protein. Cell, 1997.88(2):p.223-33), or toxins such as melittin (GIGAVLKVLTTGLPALISWIKRKRQQ, SEQ ID NO: 114) (Dempsey, CE, The actions of melittin on membranes. Biochim Biophys Acta, 1990.1031(2):p.143-61), mastoporans (Konno, K., et ah, Structure and biological activities of eumenine mastoparan-AF (EMP-AF), a new mast cell degranulating peptide in the venom of the solitary wasp (Anterhynchium flavomarginatum micado).Toxicon,2000.38(11):1505-15), maurocalcine (Esteve,E.,et al.,Transduction of the scorpion toxin maurocalcine into cells.Evidence that the toxin crosses the plasma membrane.J Biol Chem,2005.280(13):p.12833-9), crotamine (Nascimento,FD,et al.,Crotamine mediates gene delivery into cells through the binding to heparan sulfate proteoglycans.J Biol Chem,2007.282(29):p.21 349-60) or buforin (Kobayashi,S.,et al.,Membrane translocation mechanism of the antimicrobial peptide buforin 2.Biochemistry,2004.43(49):p.15610-6). Synthetic CPPs have also been designed, including polyarginine (R8, R9, R10, and R12) (Futaki, S., et al., Arginine-rich peptides. An abundant source of membrane-permeable peptides having potential as carriers for intracellular protein delivery. J Biol Chem, 2001. 276(8): p. 5836-40) or transportan (Pooga, M., et al., Cell penetration by transportan. FASEB J, 1998. 12(1): p. 67-77). Any of the above CPPs may be used in the modified Cav-1 peptides of the present disclosure. Several other CPPs described in Milletti F. (Drug Discov Today 17(15-16): 850-60, 2012) may also be used in the modified Cav-1 peptides of the present disclosure.

[0183] In some embodiments, the modified Cav-1 peptide comprises an internalization sequence. In some embodiments, the internalization sequence is located at either the C-terminus or the N-terminus of the modified Cav-1 peptide. In some embodiments, the internalization sequence comprises an amino acid sequence selected from the group comprising GRKKRRQRRRPPQ (SEQ ID NO: 112), RQIKIWFQNRRMKWKK (SEQ ID NO: 113), and GIGAVLKVLTTGLPALISWIKRKRQQ (SEQ ID NO: 114).

[0184] III. Pharmaceutical Compositions When clinical use is envisioned, it may be necessary to prepare the modified Cav-1 peptide or pharmaceutical composition thereof in a form suitable for the intended use. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises an effective amount of one or more modified Cav-1 peptides of the present disclosure dissolved or dispersed in a pharma- ceutical acceptable carrier. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises an effective amount of one or more modified Cav-1 peptides of the present disclosure and at least one additional therapeutic agent dissolved or dispersed in a pharma- ceutical acceptable carrier. The preparation of pharmaceutical compositions containing at least one modified Cav-1 peptide of the present disclosure and / or at least one additional therapeutic agent will be known to those skilled in the art in view of the present disclosure, as exemplified by Remington's Pharmaceutical Sciences, 18th Ed., 1990, which is incorporated herein by reference. Furthermore, it will be understood that for animal (e.g., human) administration, the formulation should meet the sterility, pyrogenicity, general safety, and purity standards required by the FDA's Biological Standards Division.

[0185] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises different types of carriers depending on whether the composition is administered in solid, liquid, or aerosol form and whether it needs to be sterile for the route of administration, such as injection.

[0186] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is formulated into the composition in the form of a free base, neutral, or salt. Pharmaceutically acceptable salts include acid addition salts, such as those formed with free amino groups of the proteinaceous composition, or with inorganic acids, such as hydrochloric acid or phosphoric acid, or with organic acids, such as acetic acid, oxalic acid, tartaric acid, or mandelic acid. Salts formed with free carboxyl groups can also be derived from inorganic bases, such as sodium, potassium, ammonium, calcium, or ferric hydroxide, or organic bases, such as isopropylamine, trimethylamine, histidine, or procaine. Once formulated, solutions are administered in a manner compatible with the dosage formulation and in an amount that is therapeutically effective. The formulations are easily administered in a variety of dosage forms, such as those formulated for parenteral administration, such as injection solutions or aerosols for delivery to the lungs, or those formulated for gastrointestinal administration, such as drug-releasing capsules.

[0187] In some embodiments, modified Cav-1 peptide or its pharmaceutical composition is provided in a pharma- ceutically acceptable carrier with or without inert diluent.Carrier should be assimilable, and includes liquid, semi-solid, i.e. paste, or solid carrier.Unless any conventional medium, agent, diluent, or carrier is harmful to the recipient or the therapeutic effectiveness of the composition contained therein, its use in the administrable composition for use in the practice of the present method is appropriate.Examples of carrier or diluent include fat, oil, water, saline, lipid, liposome, resin, binder, filler, etc., or combinations thereof.

[0188] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is combined with the carrier in any convenient and practical manner, i.e., by solution, suspension, emulsification, mixing, encapsulation, absorption, etc. Such procedures are routine for those skilled in the art. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is thoroughly combined or mixed with a semi-solid or solid carrier. Mixing can be performed in any convenient manner, such as grinding.

[0189] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises one or more antioxidants to retard oxidation of one or more components in the composition.Furthermore, prevention of the action of microorganisms can be achieved by preservatives, such as various antibacterial and antifungal agents, including, but not limited to, parabens (e.g., methylparaben, propylparaben), chlorobutanol, phenol, sorbic acid, thimerosal, or combinations thereof.

[0190] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises one or more stabilizers.Stabilizers can also be added to the mixing process to protect the composition from loss of therapeutic activity, such as denaturation in the stomach.Examples of stabilizers include, but are not limited to, buffers, amino acids such as glycine and lysine, carbohydrates, or lyophilization protectants such as dextrose, mannose, galactose, fructose, lactose, sucrose, maltose, sorbitol, mannitol, and the like.

[0191] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises one or more surfactants. Surfactants used according to the methods of the present disclosure include ionic and non-ionic surfactants. Representative non-ionic surfactants include anionic and non-ionic surfactants such as TWEEN®-20 and TWEEN-80® surfactants (ICI Americas, Bridgewater, NJ); poloxamers (e.g., poloxamer 188); TRITON® surfactants (Sigma, St. Louis, MO); sodium dodecyl sulfate (SDS); sodium lauryl sulfate; sodium octyl glycoside; lauryl-, myristyl-, linoleyl-, or stearyl-sulfobetaine; lauryl-, myristyl-, linoleyl-, or stearyl-sarcosine; linoleyl-, myristyl-, or cetyl-betaine; lauroamidopropyl-, cocamidopropyl-, linoleamidopropyl-, myristamidopropyl-, parylene-, cyclohexane ... myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-dimethylamine; sodium methyl cocoyl taurate or disodium oleyl methyl taurate; cationic or quaternary phospholipid surfactants such as MONAQUAT™ surfactants (Mona Industries, Inc., Paterson, NJ); polyethyl glycols; polypropyl glycols; block copolymers of ethylene and propylene glycol, such as PLURONIC® surfactants (BASF Corporation, Mount Olive, NJ); oligo(ethylene oxide) alkyl ethers; alkyl(thio)glucosides; alkyl maltosides; and phospholipids. In some embodiments, the one or more surfactants are present in the modified Cav-1 peptide or pharmaceutical composition thereof in an amount of about 0.01% to about 0.5% (weight of surfactant relative to the total weight of other solid components of the formulation, "w / w"), about 0.03% to about 0.5% (w / w), about 0.05% to about 0.5% (w / w), or about 0.1% to about 0.5% (w / w).In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is essentially free of non-ionic surfactants or essentially free of all surfactants.

[0192] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is formulated as a dry powder composition. In order for the powder to be effectively inhaled and deposited in the lungs, the particle size should generally have an aerodynamic median diameter of less than about 5 μm. In some embodiments, the dry powder formulation comprising the modified Cav-1 peptide comprises an average particle size of less than 10 μm. In some embodiments, the dry powder formulation comprising the modified Cav-1 peptide comprises an average particle size of less than 5 μm. In some embodiments, the dry powder formulation comprising the modified Cav-1 peptide comprises an average particle size of less than 1 μm. In some embodiments, the modified Cav-1 peptide comprises an average particle size of about 0.01 μm to about 10 μm, about 0.1 μm to about 8 μm, about 0.5 μm to about 7 μm, or about 1 μm to about 5 μm. In some embodiments, the dry powder formulation comprising the modified Cav-1 peptide comprises an average particle size of about 0.1 μm, about 0.5 μm, about 1 μm, about 2 μm, about 3 μm, about 4 μm, about 5 μm, about 6 μm, about 7 μm, about 8 μm, about 9 μm, about 10 μm, or any range or value therebetween. In some embodiments, 90% of the dry powder has a particle size of less than about 10 μm (Dv90 of about 10 μm). In some embodiments, the Dv90 of the dry powder is about 5 μm. The particle size of the modified Cav-1 peptide or pharmaceutical composition thereof can be reduced by any suitable method, including but not limited to, grinding, grinding, thin film freezing, spray drying, or crushing. Grinding can be performed by any method known in the art, such as air jet milling, ball milling, wet milling, media milling, high pressure homogenization, or cryogenic milling. See WO2020 / 055824, which is incorporated herein by reference in its entirety. In some embodiments, the dry powder of the modified Cav-1 peptide is substantially free of excipients. In some embodiments, the dry powder of the modified Cav-1 peptide is free of excipients. In some embodiments, the dry powder consists of the Cav-1 peptide.

[0193] The stability of the peptide after particle size reduction can be evaluated using techniques known in the art, including size exclusion chromatography, electrophoretic techniques, HPLC, mass spectrometry, spectroscopic techniques such as UV spectroscopy and circular dichroism spectroscopy, and activity (measured in vitro or in vivo). To perform an in vitro assay of protein stability, the aerosol composition can be collected and then distilled or absorbed onto a filter. To perform an in vivo assay or for pulmonary administration of the composition to a subject, a device for dispersion of dry powders is adapted for inhalation by the subject. For example, protein stability can be evaluated by determining the level of protein aggregation. In some embodiments, the dry powder composition of the modified Cav-1 peptide is substantially free of protein aggregates. The presence of soluble aggregates can be qualitatively determined using dynamic light scattering (DLS) (DynaPro-801TC, Protein Solutions, Inc., Charlottesville, VA) and / or by UV spectrophotometry.

[0194] In some embodiments, the treatment of a subject with the milled modified Cav-1 peptide comprises a regulated drug release. In some embodiments, the milled modified Cav-1 peptide is formulated for sustained or delayed release. In some embodiments, the milled modified Cav-1 peptide is formulated for fast release. In further embodiments, the milled modified Cav-1 peptide is formulated for both sustained and fast release (i.e., dual release profile).

[0195] In some embodiments, the present disclosure provides a method of administration of the inhalable modified Cav-1 peptides disclosed herein. Administration via inhalation includes, but is not limited to, the use of an inhaler or nebulizer.

[0196] In some embodiments, the inhaler is a passive dry powder inhaler (DPI), such as Plastiape RS01 single dose DPI. In a dry powder inhaler, the dry powder is stored in a reservoir and delivered to the lungs by inhalation without the use of a propellant. In some embodiments, the inhaler is a single dose DPI, such as DoseOne™, Spinhaler®, Rotohaler®, Aerolizer®, or Handihaler®. In some embodiments, the inhaler is a multi-dose DPI, such as Plastiape RS02, Turbuhaler®, Twisthaler™, Diskhaler®, Diskus®, or Ellipta™. In some embodiments, the inhaler is a multi-single dose DPI for simultaneous delivery of single doses of multiple drugs, such as Plastiape RS04 multimer dose DPI. Typically, dry powder inhalers have a drug stored in an internal reservoir, and the drug is delivered by inhalation, with or without the use of a propellant. Other types of dry powder inhalers have pre-divided doses of drug stored in capsules (e.g., cellulose or gelatin-based) or foil pouches, each of which is pierced by the device to release a dose to the patient. Dry powder inhalers may require an inhalation flow rate of more than about 30 L / min, such as between about 30 L / min and about 120 L / min, for effective delivery. In some embodiments, efficient aerosolization of milled modified Cav-1 peptide is independent of inhalation force. In some embodiments, dry powder inhalers require an inhalation flow rate of more than about 0.01 kPa. 0.5 min / L and approx. 0.05 kPa 0.5 min / L, for example, about 0.02 kPa0. 5 min / L and approximately 0.04 kPa0. 5 min / L, etc. The dry powder inhaler (e.g., high resistance, low resistance, passive, active) is selected based on the patient population and their inhalation capabilities.

[0197] In some embodiments, the inhaler may be a metered dose inhaler. Metered dose inhalers deliver a defined amount of medicament to the lungs in a short burst of aerosolized medicament assisted by the use of a propellant. Metered dose inhalers include three main parts: a canister, a metered dose valve, and an actuator, and may utilize a spacer device to slow down the emitted particles and allow the patient to easily inhale the aerosolized cloud. The pharmaceutical formulation is stored in the canister, including the propellant and any necessary excipients. The metered dose valve allows for the dispensing of a defined amount of the pharmaceutical formulation. The actuator of the metered dose inhaler or mouthpiece includes a mating discharge nozzle, typically with a dust cap to prevent contamination. In some embodiments, the inhalation flow rate required for use of the metered dose inhaler is less than about 90 L / min, for example between about 15 L / min and about 90 L / min, preferably about 30 L / min. In some embodiments, efficient aerosolization of milled modified Cav-1 peptide is independent of inhalation force.

[0198] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is delivered to a subject by inhalation through the use of a nebulizer. A nebulizer is used to deliver a pharmaceutical in the form of an aerosolized mist that is inhaled into the lungs. The pharmaceutical formulation is aerosolized by compressed gas or by ultrasound. The jet nebulizer is connected to a compressor. The compressor releases compressed gas through the liquid pharmaceutical formulation at high speed, thereby aerosolizing the pharmaceutical formulation. The aerosolized pharmaceutical is then inhaled by the patient. The ultrasonic nebulizer generates high-frequency ultrasound, causing vibration of an internal element that contacts the liquid reservoir of the pharmaceutical formulation, thereby aerosolizing the pharmaceutical formulation. The aerosolized pharmaceutical is then inhaled by the patient. The nebulizer may utilize a flow rate between about 3L / min and about 12L / min, such as about 6L / min. In some examples, the milled modified Cav-1 peptide is suspended in a pharma-ceutically acceptable liquid carrier vehicle and can be administered by nebulization (e.g., air jet nebulization). In some embodiments, the modified Cav-1 peptide is administered by a vaporization method (e.g., rapid vaporization), such as an e-cigarette device.

[0199] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is nebulized into aerosol droplets. In some embodiments, the aerosol droplets have a median diameter of about 1 μm to about 20 μm. In some embodiments, the aerosol droplets have a median diameter of about 2.5 μm to about 20 μm. In some embodiments, the aerosol droplets have a median diameter of about 2 μm to about 10 μm. In some embodiments, the aerosol droplets have a median diameter of about 2 μm to about 4 μm. In some embodiments, the aerosol droplets have a median diameter of about 1 μm to about 5 μm. In some embodiments, the aerosol droplets have a median diameter of about 0.5 μm, about 1 μm, about 2 μm, about 3 μm, about 4 μm, about 5 μm, about 6 μm, about 7 μm, about 8 μm, about 9 μm, about 10 μm, about 11 μm, about 12 μm, about 13 μm, about 14 μm, about 15 μm, about 16 μm, about 17 μm, about 18 μm, about 19 μm, about 20 μm, or any size in between.

[0200] IV.How to use The present disclosure provides uses of modified Cav-1 peptides or pharmaceutical compositions thereof described herein. Specifically, these methods relate to administering any one of the modified Cav-1 peptides or pharmaceutical compositions thereof described herein to a subject. In some embodiments, the modified Cav-1 peptides or pharmaceutical compositions thereof are used to treat or prevent a kidney disease or disorder (e.g., chronic kidney disease) in a subject. In some embodiments, the modified Cav-1 peptides or pharmaceutical compositions thereof are used to treat or prevent a lung disease or disorder (e.g., idiopathic pulmonary fibrosis) in a subject. In some embodiments, the subject is elderly.

[0201] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is used to treat or prevent kidney disease or disorder (e.g., chronic kidney disease) in a subject. In some embodiments, the modified Cav-1 peptide is used to treat or prevent kidney disease or disorder, such as, for example, chronic kidney disease, end-stage renal disease, glomerulonephritis, focal segmental glomerulosclerosis, renal fibrosis, polycystic kidney disease, IgA nephropathy, lupus nephritis, nephrotic syndrome, Alport syndrome, amyloidosis, Goodpasture's syndrome, Wegener's granulomatosis, or acute kidney injury. In some embodiments, the kidney disease is characterized by fibrosis. In some embodiments, the kidney disease or disorder is acute. In some embodiments, the kidney disease or disorder is chronic. In some embodiments, the subject is elderly. In some embodiments, the kidney disease or disorder in the subject is caused by infection, such as, for example, a viral, bacterial, fungal, or parasitic infection. In some embodiments, the kidney disease or disorder in the subject is caused by hypertension (elevated blood pressure). In some embodiments, the kidney disease or disorder in the subject is caused by diabetes.In some embodiments, the kidney disease or disorder in the subject is caused by overuse of drugs such as OTC painkillers or heroin.In some embodiments, the subject has high blood pressure or diabetes.

[0202] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is used to delay the progression of kidney disease or disorder in a subject. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is used to improve progression-free survival. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is used to prolong the survival of a subject.

[0203] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent chronic kidney disease in a subject.Chronic kidney disease is the gradual and progressive loss of kidney's ability to excrete waste products, concentrate urine, and conserve electrolytes.Progressive loss of kidney function occurs as a result of the deposition of fibrous tissue between the functional units of the kidney or nephron (interstitial fibrosis) and the continuous replacement of filtering surface by fibrous tissue (glomerulosclerosis).Renal fibrosis is the pathological characteristic of chronic kidney disease and is the main contributing factor of progression to end-stage renal disease.In one embodiment, the chronic kidney disease is chronic kidney fibrosis.

[0204] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent end-stage renal disease in a subject.End-stage renal disease is the final stage of chronic kidney disease, when the kidneys stop functioning and the individual requires long-term dialysis or kidney transplantation to survive.

[0205] In some embodiments, the modified Cav-1 peptides of the present disclosure are used to treat or prevent focal segmental glomerulosclerosis in a subject.Focal segmental glomerulosclerosis is a kidney disease characterized by scarring of the glomeruli, which causes protein loss in the urine.

[0206] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent glomerulonephritis in a subject.Glomerulonephritis, also known as glomerular disease, is a type of kidney disease in which glomeruli are damaged and cannot properly remove waste and fluid from the body.In some embodiments, the glomerulonephritis is acute glomerulonephritis.In some embodiments, the glomerulonephritis is chronic glomerulonephritis.

[0207] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent polycystic kidney disease in a subject.Polycystic kidney disease is a genetic disorder in which clusters of cysts develop, primarily in the kidney, causing the kidney to enlarge and lose function over time.

[0208] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent IgA nephropathy in a subject. IgA nephropathy, also known as Berge's disease, is a kidney disease that occurs when immunoglobulin IgA accumulates in the kidney, resulting in local inflammation that can prevent the kidney from filtering waste products from the blood.

[0209] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent lupus nephritis in a subject. Lupus nephritis constitutes one of the most severe organ manifestations of systemic lupus erythematosus and occurs when the immune system attacks the kidney. It is a type of glomerulonephritis.

[0210] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent nephrotic syndrome in a subject.Nephrotic syndrome is a kidney disorder that causes the body to excrete too much protein in urine.Nephrotic syndrome is often caused by damage to the small blood vessels in the kidney that filter waste products and excess water from the blood.

[0211] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent Alport syndrome in a subject. Alport syndrome is a genetic condition characterized by progressive kidney disease and abnormalities of the inner ear and eyes. There are three genetic types: X-linked Alport syndrome (XLAS), autosomal recessive Alport syndrome (ARAS), and autosomal dominant Alport syndrome (ADAS). XLAS is caused by mutations in the COL4A5 gene, ARAS is caused by mutations in both copies of either the COL4A3 gene or the COL4A4 gene, and ADAS is caused by mutations in one copy of the COL4A3 gene or the COL4A4 gene. Individuals with Alport syndrome exhibit chronic glomerular dysfunction, renal inflammation, and fibrosis, which are characteristic of chronic kidney disease, and progress to end-stage renal disease. People affected by Alport syndrome may also develop progressive hearing loss of various severity and eye abnormalities that do not usually result in vision problems.

[0212] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent amyloidosis in a subject. Amyloidosis occurs when amyloid accumulates in tissues and organs and interferes with normal function. Amyloid deposits damage the kidneys and affect their ability to filter waste and break down proteins. In some embodiments, the amyloidosis is primary amyloidosis. In some embodiments, the amyloidosis is dialysis-associated amyloidosis.

[0213] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent Goodpasture's syndrome in a subject.Goodpasture's syndrome, also known as anti-glomerular basement membrane disease, is an autoimmune disease in which antibodies attack the basement membrane of the lungs and kidneys, leading to pulmonary hemorrhage, glomerulonephritis, and kidney failure.

[0214] In some embodiments, modified Cav-1 peptides of the present disclosure are used to treat or prevent granulomatosis with polyangiitis in a subject. Granulomatosis with polyangiitis, also known as Wegener's granulomatosis, is an autoimmune disease involving granulomatous inflammation, necrosis, and vasculitis that most frequently targets the lungs and kidneys.

[0215] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent acute kidney injury in a subject.Acute kidney injury, also called acute renal failure, is the sudden loss of renal excretory function.Acute kidney injury is defined by serum creatine and urinary excretion levels for less than one week.

[0216] In some embodiments, the modified Cav-1 peptide is used to treat or prevent kidney infection. In some embodiments, the modified Cav-1 peptide is used to treat or prevent pyelonephritis. Pyelonephritis is a type of urinary tract infection in which one or both kidneys become infected. In some embodiments, pyelonephritis is caused by bacteria or viruses, such as Escherichia coli, Klebsiella, Proteus, Pseudomonas, Enterococcus, or Staphylococcus saprophyticus.

[0217] In some embodiments, the modified Cav-1 peptide is used to treat or prevent a kidney disease or disorder resulting from a microbial infection in a subject. In some embodiments, the microbial infection is a bacterial, viral, fungal, or parasitic infection. In some embodiments, the kidney disease or disorder is caused by Streptococcus pyogenes, Staphylococcus aureus, Epidermidis, Salmonella (typhi, paratyphi), Escherichia coli, Caused by Leptospira spp., Mycobacterium tuberculosis, Mycobacterium leprae, Legionella spp., Yersinia enterocolitica, Brucella spp., Campylobacter jejuni, Corynebacterium diphtheriae, Klebsiella spp., Proteus spp., Pseudomonas spp., Enterococcus spp., Staphylococcus saprophyticus, SARS-CoV-1, SARS-CoV-2, Dengue virus, Hantavirus, Varicella zoster virus, Parvovirus, Hepatitis A virus, Hepatitis B virus, Hepatitis E virus, Cytomegalovirus, Epstein-Barr virus, Human immunodeficiency virus, and / or Hepatitis C virus.

[0218] In some embodiments, the modified Cav-1 peptides are used to treat or prevent kidney disease or injury resulting from SARS-CoV-2 infection. In some embodiments, SARS-CoV-2 causes acute kidney injury. In some embodiments, SARS-CoV-2 causes chronic kidney injury. In some embodiments, SARS-CoV-2 causes chronic kidney disease. In some embodiments, SARS-CoV-2 causes renal fibrosis. In some embodiments, SARS-CoV-2 causes renal failure.

[0219] In some embodiments, the disclosure provides a method of treating or preventing a kidney disease or disorder in a subject, the method comprising administering to the subject an effective amount of a modified Cav-1 peptide or a pharmaceutical composition thereof. In some embodiments, the disclosure provides a method of treating or preventing a kidney disease or disorder in a subject, the method comprising administering to the subject an effective amount of a modified Cav-1 peptide comprising an amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the disclosure provides a method of treating or preventing a kidney disease or disorder in a subject, the method comprising administering to the subject an effective amount of a modified Cav-1 peptide comprising an amino acid sequence of any one of SEQ ID NOs: 4-20. In some embodiments, the disclosure provides a method of treating or preventing a kidney disease or disorder in a subject, the method comprising administering to the subject an effective amount of a modified Cav-1 peptide comprising an amino acid sequence of SEQ ID NO: 3. In some embodiments, the disclosure provides a method of treating or preventing a kidney disease or disorder in a subject, the method comprising administering to the subject an effective amount of a modified Cav-1 peptide comprising an amino acid sequence of SEQ ID NO: 8. In some embodiments, the disclosure provides a method of treating or preventing a kidney disease or disorder in a subject, the method comprising administering to the subject an effective amount of a modified Cav-1 peptide comprising at least one amino acid substitution, insertion or deletion relative to the amino acid sequence FTTFTVT (SEQ ID NO: 3), wherein the modified Cav-1 peptide maintains the biological activity of Cav-1. In some embodiments, the kidney disease or disorder is chronic kidney disease. In some embodiments, the kidney disease or disorder is Alport syndrome.

[0220] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is used to improve renal function in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof improves renal function by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% compared to the subject prior to treatment with the modified Cav-1 peptide or pharmaceutical composition thereof. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises any one of the amino acid sequences of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 8. In some embodiments, improved renal function refers to a reduction in fibrotic glomeruli, a reduction in blood urea nitrogen, a reduction in blood creatinine, an increase in blood albumin, a reduction in the urinary albumin to creatinine ratio, and / or an increase in glomerular filtration rate.

[0221] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces the number of fibrotic glomeruli in a subject with a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces the number of fibrotic glomeruli by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% compared to the subject prior to treatment with the modified Cav-1 peptide or pharmaceutical composition thereof. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 8.

[0222] In some embodiments, a subject with a renal disease or disorder has reduced expression of caveolin-1 in the kidney compared to a subject without a renal disease or disorder (e.g., a normal, healthy subject). In some embodiments, expression of caveolin-1 is reduced in glomeruli of a subject with a renal disease or disorder compared to a subject without a renal disease or disorder. In some embodiments, expression of caveolin-1 is reduced in renal endothelial cells of a subject with a renal disease or disorder compared to a subject without a renal disease or disorder. In some embodiments, expression of caveolin-1 is reduced in renal epithelial cells of a subject with a renal disease or disorder compared to a subject without a renal disease or disorder. In some embodiments, expression of caveolin-1 is reduced in renal podocytes of a subject with a renal disease or disorder compared to a subject without a renal disease or disorder. In some embodiments, expression of caveolin-1 in the kidney is reduced by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% in a subject with a renal disease or disorder compared to a subject without a renal disease or disorder. In some embodiments, the epithelial cells are parietal epithelial cells lining Bowman's capsule.

[0223] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces endothelial cell death in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces epithelial cell death in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces podocyte cell death in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces cell death by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% compared to the subject prior to treatment with the modified Cav-1 peptide or pharmaceutical composition thereof. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 8.

[0224] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases endothelial cell viability in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases epithelial cell viability in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases podocyte viability in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases cell viability by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% compared to the subject prior to treatment with the modified Cav-1 peptide or pharmaceutical composition thereof. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 8.

[0225] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof promotes kidney regeneration in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof promotes regeneration of renal blood vessels, glomeruli, and / or tubules in the kidney of a subject. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof promotes regeneration of epithelial cells, endothelial cells, tubular cells, and / or podocytes in the kidney of a subject. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 8.

[0226] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases endothelial cell proliferation in a subject having a renal disease or disorder. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases epithelial cell proliferation in a subject having a renal disease or disorder. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases podocyte proliferation in a subject having a renal disease or disorder. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases cell proliferation by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% compared to the subject prior to treatment with the modified Cav-1 peptide or pharmaceutical composition thereof. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 8.

[0227] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces blood urea nitrogen in a subject with kidney disease or disorder. In some embodiments, high blood urea nitrogen values ​​indicate kidney damage or disease in the subject. Generally, blood urea nitrogen levels in the range of 6 mg / dl to 24 mg / dl are considered normal in humans. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces blood urea nitrogen by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% compared to the subject before treatment with the modified Cav-1 peptide or pharmaceutical composition thereof. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces blood urea nitrogen in a subject to less than about 50 mg / dl, less than about 45 mg / dl, less than about 40 mg / dl, less than about 35 mg / dl, less than about 30 mg / dl, less than about 25 mg / dl, or less than about 20 mg / dl. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces blood urea nitrogen to less than about 20 mg / dl. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises an amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises an amino acid sequence of SEQ ID NO: 3. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises an amino acid sequence of SEQ ID NO: 8.

[0228] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces blood creatinine in a subject with kidney disease or disorder. In some embodiments, high blood creatinine levels indicate kidney damage or disease in the subject. In general, blood creatinine levels above 1.2 mg / dl in women and above 1.4 mg / dl in men indicate that the kidneys are not functioning properly. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces blood creatinine by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% compared to the subject before treatment with the modified Cav-1 peptide or pharmaceutical composition thereof. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces blood creatinine in the subject to less than about 4 mg / dl, less than about 3.5 mg / dl, less than about 3.25 mg / dl, less than about 3 mg / dl, less than about 2.75 mg / dl, less than about 2.5 mg / dl, less than about 2.25 mg / dl, less than about 2 mg / dl, less than about 1.75 mg / dl, less than about 1.5 mg / dl, or less than about 1.25 mg / dl. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces blood urea nitrogen to less than about 1.5 mg / dl. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 8.

[0229] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases blood albumin in a subject with kidney disease or disorder. In some embodiments, low blood albumin levels may indicate kidney damage or disease in a subject. Normal levels of albumin in blood are 3.5 g / dL to 5 g / dL. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases blood albumin by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% compared to the subject prior to treatment with the modified Cav-1 peptide or pharmaceutical composition thereof. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases blood albumin in the subject to greater than about 1.5 g / dl, greater than about 1.75 g / dl, greater than about 2 g / dl, greater than about 2.25 g / dl, greater than about 2.5 g / dl, greater than about 2.75 g / dl, greater than about 3 g / dl, or greater than about 3.5 g / dl. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases blood albumin to greater than about 3.5 g / dl. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 8.

[0230] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces the urinary albumin to creatinine ratio in a subject with a kidney disease or disorder. The urinary albumin to creatinine ratio is useful for identifying kidney damage or disease in a subject. An albumin to creatinine ratio of less than 30 mg / g is considered normal, a ratio of 30-300 mg / g indicates microalbuminuria, and values ​​above 300 mg / g indicate microalbuminuria in humans. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces the urinary albumin to creatinine ratio by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% compared to the subject prior to treatment with the modified Cav-1 peptide or pharmaceutical composition thereof. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces the urinary albumin to creatinine ratio in the subject to less than about 300 mg / g, less than about 250 mg / g, less than about 200 mg / g, less than about 150 mg / g, less than about 100 mg / g, less than about 75 mg / g, less than about 50 mg / g, less than about 40 mg / g, less than about 30 mg / g, or less than about 25 mg / dl. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof reduces the urinary albumin to creatinine ratio to less than about 30 mg / dl. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises any one of the amino acid sequences of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 8.

[0231] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases the glomerular filtration rate in a subject with a kidney disease or disorder. The glomerular filtration rate indicates how well the kidney filters blood to remove waste and excess water to make urine. 90 mL / min / 1.73 m 2A glomerular filtration rate of more than 60 mL / min / 1.73 m is considered normal, whereas a glomerular filtration rate of more than 60 mL / min / 1.73 m is considered normal. 2 A glomerular filtration rate of less than 15 mL / min / 1.73 m can indicate renal injury or disease. 2 A glomerular filtration rate of less than about 30 mL / min / 1.73 m may be indicative of renal failure. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases the glomerular filtration rate by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 100% compared to the subject prior to treatment with the modified Cav-1 peptide or pharmaceutical composition thereof. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases the glomerular filtration rate in the subject by at least about 30 mL / min / 1.73 m 2 Super, about 40mL / min / 1.73m 2 Super, about 50mL / min / 1.73m 2 Super, about 60mL / min / 1.73m 2 Super, about 70mL / min / 1.73m 2 Super, about 80mL / min / 1.73m 2 More than or about 90mL / min / 1.73m 2 In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof increases the glomerular filtration rate in a subject to about 60 mL / min / 1.73 m 2 In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 8.

[0232] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is used to preserve renal function in a subject with renal disease or disorder. As used herein, the term "preserve" refers to the maintenance of renal function or the prevention of further decline in renal function in a subject with renal disease or disorder. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof preserves renal function as measured by blood urinary nitrogen, blood creatinine, blood albumin, urinary albumin to creatinine ratio, and / or glomerular filtration rate, which measurements remain stable upon treatment with the modified Cav-1 peptide or pharmaceutical composition thereof. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof comprises the amino acid sequence of SEQ ID NO: 8.

[0233] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is used to treat or prevent a disease or disorder in an elderly subject. As used herein, the term "elderly" or "elderly" refers to a subject aged 55 years or older. In some embodiments, the elderly subject is about 55 years old, about 60 years old, about 65 years old, about 70 years old, about 75 years old, about 80 years old, about 90 years old, about 95 years old, or about 100 years old. In some embodiments, the elderly subject is more susceptible to a disease or disorder described herein than a younger subject. In some embodiments, the elderly subject has a fibrotic disease or disorder, such as idiopathic pulmonary fibrosis.

[0234] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is used to treat or prevent fibrotic diseases or disorders in elderly subjects. In some embodiments, the modified Cav-1 peptide is used to treat or prevent fibrotic diseases or disorders, such as interstitial lung disease, liver fibrosis, renal fibrosis, skin fibrosis, glomerulonephritis, systemic sclerosis, cardiac fibrosis, myocardial fibrosis, renal fibrosis, liver cirrhosis, nephrosclerosis, arteriosclerosis, macular degeneration, ocular scarring, cataracts, retinal and vitreous retinopathy, Graves' ophthalmopathy, neurofibromatosis, scleroderma, glioblastoma, keloids and hypertrophic scarring, peritoneal fibrotic disease, chronic obstructive pulmonary disease, postoperative fibroids, diabetic nephropathy, gynecological cancer, myeloproliferative disorders, myeloid leukemia, myelodysplastic syndrome, inflammatory bowel disease, nonalcoholic fatty liver disease, fibrosarcoma, rheumatoid arthritis, nonalcoholic steatohepatitis, Alport syndrome, or chronic COVID syndrome.

[0235] In some embodiments, an elderly subject has reduced expression of caveolin-1 compared to a younger subject (e.g., a young adult or middle-aged subject), hi some embodiments, expression of caveolin-1 is reduced by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% in an elderly subject compared to a younger subject.

[0236] In some embodiments, the present disclosure provides a method for treating or preventing a fibrotic disease or disorder in an elderly subject, the method comprising administering to the elderly subject an effective amount of a modified Cav-1 peptide or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method for treating or preventing a fibrotic disease or disorder in an elderly subject, the method comprising administering to the elderly subject an effective amount of a modified Cav-1 peptide comprising an amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the present disclosure provides a method for treating or preventing a fibrotic disease or disorder in an elderly subject, the method comprising administering to the elderly subject an effective amount of a modified Cav-1 peptide comprising an amino acid sequence of any one of SEQ ID NOs: 4-20. In some embodiments, the present disclosure provides a method for treating or preventing a fibrotic disease or disorder in an elderly subject, the method comprising administering to the elderly subject an effective amount of a modified Cav-1 peptide comprising an amino acid sequence of SEQ ID NO: 3. In some embodiments, the present disclosure provides a method for treating or preventing a fibrotic disease or disorder in an elderly subject, the method comprising administering to the elderly subject an effective amount of a modified Cav-1 peptide comprising an amino acid sequence of SEQ ID NO: 8. In some embodiments, the disclosure provides a method of treating or preventing a fibrotic disease or disorder in an elderly subject, the method comprising administering to the elderly subject an effective amount of a modified Cav-1 peptide comprising at least one amino acid substitution, deletion, insertion relative to the amino acid sequence FTTFTVT (SEQ ID NO: 3), wherein the modified Cav-1 peptide maintains the biological activity of Cav-1.

[0237] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is used to treat or prevent a pulmonary disease or disorder in a subject. In some embodiments, the modified Cav-1 peptide is used to treat or prevent a pulmonary disease or disorder, such as, for example, acute lung injury (ALI), chronic lung injury, acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD), asthma, interstitial lung disease, pulmonary fibrosis, pneumonia, hypersensitivity interstitial pneumonia, bronchiolitis, sarcoidosis, scleroderma, or pulmonary infection. In some embodiments, the modified Cav-1 peptide is used to treat or prevent a pulmonary disease or disorder in an elderly subject. In some embodiments, the elderly subject has an interstitial lung disease, such as idiopathic pulmonary fibrosis.

[0238] In some embodiments, the modified Cav-1 peptides of the present disclosure are used to treat or prevent a pulmonary infection in a subject, such as a bacterial infection, a viral infection, or a fungal infection. In some embodiments, the pulmonary infection causes one or more pulmonary diseases or disorders in the subject, including, but not limited to, ALI, ARDS, COPD, asthma, interstitial lung disease, pulmonary fibrosis, pneumonia, hypersensitivity interstitial pneumonia, bronchiolitis, sarcoidosis, and scleroderma.

[0239] In some embodiments, the modified Cav-1 peptides of the present disclosure are used to treat or prevent bacterial infections in a subject. Examples of bacteria that cause lung infections include, but are not limited to, Pseudomonas aeruginosa, Bacillus anthracis, Listeria monocytogenes, Staphylococcus aureus, Streptococcus pneumoniae, Haemophilus influenzae, Enterobacteriaceae, Nocardia, Actinomyces, Moraxella catarrhalis, Klebsiella pneumoniae, Chlamydia trachomatis, Chlamydophila pneumoniae, Chlamydophila psittaci, Coxiella burnetii, Salmonella, Yersinia pestis, Mycobacterium leprae, Mycobacterium africanum, Mycobacterium asiaticum, Mycobacterium avium-cellulos ... In some embodiments, the bacterial infection causes pneumonia in the subject.

[0240] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent viral infection in a subject. In some embodiments, the modified Cav-1 peptide is used to treat or prevent infection in a subject caused by double-stranded DNA (dsDNA) virus, single-stranded DNA (ssDNA) virus, single-stranded RNA (ssRNA) virus, or double-stranded RNA (dsRNA) virus. In some embodiments, the ssRNA virus is a positive-sense ssRNA virus (+ssRNA). In some embodiments, the ssRNA virus is a negative-sense ssRNA virus (-ssRNA). Examples of viruses that cause lung infections include, but are not limited to, coronaviruses (e.g., SARS-CoV-1, SARS-CoV-2, or MERS-CoV), influenza, respiratory syncytial virus, metapneumovirus, bocavirus, parainfluenza, rhinovirus, enterovirus, norovirus, adenovirus, varicella zoster virus, hantavirus, parechovirus, Epstein-Barr virus, herpes simplex virus, mimivirus, cytomegalovirus, torque teno virus, and Middle East respiratory syndrome coronavirus. In some embodiments, the viral infection causes pneumonia in the subject. In some embodiments, the viral infection causes pulmonary fibrosis in the subject. In some embodiments, the viral infection causes bronchiolitis in the subject. In some embodiments, the viral infection causes ALI or ARDS in the subject. In some embodiments, the viral infection causes interstitial lung disease in the subject. In some embodiments, the viral infection causes asthma in the subject. In some embodiments, the viral infection causes sarcoidosis in the subject. In some embodiments, the viral infection causes scleroderma in the subject.

[0241] In some embodiments, SARS-CoV-1 causes Severe Acute Respiratory Syndrome (SARS) in a subject. In some embodiments, modified Cav-1 peptides are used to treat or prevent SARS in a subject. SARS is characterized by initial systemic symptoms of muscle pain, headache, and fever, followed by respiratory symptoms, primarily cough, dyspnea, and pneumonia, over a period of 2-14 days.

[0242] In some embodiments, MERS-CoV causes Middle East Respiratory Syndrome (MERS) in a subject. In some embodiments, modified Cav-1 peptides are used to treat or prevent MERS in a subject. The clinical features of MERS range from asymptomatic or mild disease to acute respiratory distress syndrome and multiple organ failure, the latter of which can result in death, especially in individuals with underlying diseases. There is no specific drug treatment for MERS, and infection prevention and control measures are important to prevent spread in health care facilities. See Zumla et al. Lancet 2015;386(9997):995-1007.

[0243] In some embodiments, SARS-CoV-2 causes coronavirus disease 2019 (COVID-19) in a subject. In some embodiments, the modified Cav-1 peptide is used to treat or prevent an infection caused by SARS-CoV-2. In some embodiments, a variant of SARS-CoV-2 causes COVID-19 in a subject. In some embodiments, the modified Cav-1 peptide is used to treat or prevent an infection caused by a variant of SARS-CoV-2. In some embodiments, the modified Cav-1 peptide is used to treat or prevent an infection caused by a SARS-CoV-2 alpha variant. In some embodiments, the modified Cav-1 peptide is used to treat or prevent an infection caused by a SARS-CoV-2 beta variant. In some embodiments, the modified Cav-1 peptide is used to treat or prevent an infection caused by a SARS-CoV-2 gamma variant. In some embodiments, the modified Cav-1 peptide is used to treat or prevent an infection caused by a SARS-CoV-2 delta variant. In some embodiments, the modified Cav-1 peptides are used to treat or prevent an infection caused by a SARS-CoV-2 epsilon variant. In some embodiments, the modified Cav-1 peptides are used to treat or prevent an infection caused by a SARS-CoV-2 zeta variant. In some embodiments, the modified Cav-1 peptides are used to treat or prevent an infection caused by a SARS-CoV-2 eta variant. In some embodiments, the modified Cav-1 peptides are used to treat or prevent an infection caused by a SARS-CoV-2 theta variant. In some embodiments, the modified Cav-1 peptides are used to treat or prevent an infection caused by a SARS-CoV-2 iota variant. In some embodiments, the modified Cav-1 peptides are used to treat or prevent an infection caused by a SARS-CoV-2 kappa variant.In some embodiments, the modified Cav-1 peptides are used to treat or prevent infection caused by SARS-CoV-2 lambda variants. In some embodiments, the modified Cav-1 peptides are used to treat or prevent infection caused by SARS-CoV-2 mu variants. In some embodiments, the modified Cav-1 peptides are used to treat or prevent infection caused by SARS-CoV-2 omicron variants. In some embodiments, the SARS-CoV-2 variant is B.1.1.7 (also known as 501Y.V1 or VOC-202012 / 01), B.1.1.317, B.1.1.318, B.1.1.529, B.1.351 (also known as 501Y.V2), B.1.429, B1.427, B1.1.207, A.23.1, COH.20G / 501Y, B.1.525, B.1.526, B.1.617, B.1.618, B.1.621, C.37, P.1, P.2, or P.3, or subvariants thereof, or combinations thereof. See Konings et al., Variants of Interest and Concern naming scheme conducive for global discourse. Nature Microbiology (2021). In some embodiments, the subvariant of the SARS-CoV-2 variant B.1.1.529 is BA.1 (B1.1.529.1), BA1.1 (B1.1.529.1.1), BA.2 (B1.1.529.2), BA.3 (B1.1.529.3), BA.4 (B1.1.529.4), or BA.5 (B1.1.529.5). In some embodiments, the subvariant of the SARS-CoV-2 variant B.1.1.7 is Q.1, Q.2, Q.3, Q.4, Q.5, Q.6, Q.7, or Q.8. In some embodiments, the subvariant of the SARS-CoV-2 variant B.1.351 is B.1.351.1, B.1.351.2, B.1.351.3, B.1.351.4, or B.1.351.5.In some embodiments, the sub-variant of the SARS-CoV-2 variant P.1 is P.1.1, P.1.2, P.1.3, P.1.4, P.1.5, P.1.6, P.1.7, P.1.7.1, P.1.8, P.1.9, P.1.10, P.1.10.1, P.1.10.2, P.1.11, P.1.12, P.1.12.1, P.1.13, P.1.14, P.1.15, P.1.16, P.1.17, or P.1.17.1. In some embodiments, the sub-variant of the SARS-CoV-2 variant B.1.617 is B.1.617.1, B.1.617.2, or B.1.617.3. In some embodiments, the sub-variant of the SARS-CoV-2 variant B.1.526 is B.1.526.1. In some embodiments, the sub-variant of the SARS-CoV-2 variant B.1.621 is B.1.621.1, B.1.621.2, BB.1, or BB.2. In some embodiments, the sub-variant of the SARS-CoV-2 variant C.37 is C.37.1.

[0244] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent fungal infection in a subject. Examples of fungi that cause lung infection include, but are not limited to, Candida (e.g., Candida albicans, Candida glabrata, Candida krusei), Aspergillus, Pneumocystis, Coccidioides (e.g., Coccidioides immitis, Coccidioides posadaci), Brustomyces (e.g., Brustomyces dermatitidis), Histoplasma (e.g., Histoplasma capsulatum), Cryptococcus (e.g., Cryptococcus neoformans, Cryptococcus gattii), Sporothrix (e.g., Sporothrix schenckii), Mucor, and Paracoccidioides. In some embodiments, the fungal infection causes pneumonia in the subject. In some embodiments, fungal infection causes invasive pulmonary aspergillosis in the subject. In some embodiments, fungal infection causes allergic asthma, allergic bronchopulmonary aspergillosis, or hypersensitivity interstitial pneumonia in the subject. In some embodiments, fungal infection causes ARDS. In some embodiments, fungal infection causes pulmonary fibrosis in the subject. In some embodiments, fungal infection causes pulmonary edema in the subject.

[0245] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent interstitial lung disease in a subject.Interstitial lung disease is a group of disorders that cause interstitial fibrosis and inflammation.In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis, lymphangioleiomyomatosis, nonspecific interstitial pneumonia, idiopathic interstitial pneumonia, cryptogenic organizing pneumonia, acute interstitial pneumonia, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, lymphocytic interstitial pneumonia, pulmonary sarcoidosis, diffuse alveolar damage, systemic sclerosis, polymyositis, systemic lupus erythematosus, rheumatoid arthritis, drug-induced interstitial lung disease, or occupational interstitial lung disease.In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0246] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent acute lung injury (ALI) in a subject. ALI is an acute inflammatory disorder that causes the destruction of endothelial and epithelial barriers in the lung. ALI can be the result of inhalation injury or the result of a systemic disease such as sepsis or severe hypovolemic shock. In some embodiments, the ALI is chemically induced ALI. In some embodiments, the ALI is inhalation smoke-induced acute lung injury (ISALI). In some embodiments, the ALI is ARDS.

[0247] In some embodiments, the modified Cav-1 peptide of the present disclosure is used to treat or prevent ARDS in a subject.ARDS is the most severe form of ALI and is distinguished by the severity of oxygen deficiency.ARDS is a life-threatening type of lung injury that occurs when fluid accumulates in the elastic small air sacs (alveoli) of the lungs.Fluid in the alveoli prevents the lungs from filling with oxygen, resulting in less oxygen reaching the bloodstream and making breathing difficult.

[0248] In some embodiments, modified Cav-1 peptides are used to treat or prevent cystic fibrosis (CF) in a subject. CF is an inherited disease of the exocrine and exocrine sweat glands that primarily affects the digestive and respiratory systems. The disease is usually characterized by chronic respiratory infections, pancreatic insufficiency, abnormally viscous mucus secretion, and premature death. CF is characterized by progressive airflow obstruction. A subset of individuals with CF also develops airway hyperresponsiveness to inhaled cholinergic agents (Weinberger, 2002 and Mitchell et al., 1978) and reversible airflow limitation in response to bronchodilators (van Haren et al., 1991 and van Haren et al., 1992). The presence of bronchial hyperresponsiveness and airway obstruction suggests that disease pathogenesis may be shared between CF and other diseases of airway narrowing, such as asthma and COPD, and airway smooth muscle dysfunction is thought to contribute to these disease processes.

[0249] In some embodiments, modified Cav-1 peptides are used to treat or prevent COPD in subjects. COPD is a term used to classify two major airflow obstruction disorders: chronic bronchitis and emphysema. Chronic bronchitis is inflammation of the bronchial airways. Bronchial airways connect the trachea to the lungs. When inflamed, the bronchial airways secrete mucus, causing chronic coughing. In emphysema, damage to the elastin framework of the lung results in hyperinflation of the alveolar sacs. Inflammatory cells in emphysematous lungs release elastase enzymes, which break down or damage elastase fibers in the lung matrix. Emphysema has many causes, including smoking, exposure to environmental pollutants, alpha-1 antitrypsin deficiency, and aging.

[0250] In some embodiments, the modified Cav-1 peptide disclosed herein is used to treat or prevent bronchiolitis in a subject. Bronchiolitis is most commonly caused by viral lower respiratory tract infections and is characterized primarily by acute inflammation, edema, necrosis of epithelial cells lining small airways, and increased mucus production (Ralston et al., 2014). Signs and symptoms typically begin with rhinitis and coughing, which may progress to tachypnea, wheezing, rales, accessory muscle use, and / or nasal flaring.

[0251] In some embodiments, modified Cav-1 peptides disclosed herein are used to treat or prevent bronchiolitis obliterans in subjects. Bronchiolitis obliterans is a progressive airflow reduction resulting from abnormal remodeling of small airways in the lungs (Meyer et al., 2014). Bronchiolitis obliterans is a major complication of lung transplantation and is often used to explain delayed allograft dysfunction, resulting in persistent decline in forced expiratory volume and power that is not caused by other known causes (Meyer et al., 2014).

[0252] In some embodiments, modified Cav-1 peptides disclosed herein are used to treat or prevent asthma in subjects.The term "asthma" can refer to acute asthma, chronic asthma, intermittent asthma, mild persistent asthma, moderate persistent asthma, severe persistent asthma, chronic persistent asthma, mild to moderate asthma, mild to moderate persistent asthma, mild to moderate chronic persistent asthma, allergic (extrinsic) asthma, non-allergic (intrinsic) asthma, nocturnal asthma, bronchial asthma, exercise-induced asthma, occupational asthma, seasonal asthma, silent asthma, gastroesophageal asthma, idiopathic asthma, and cough asthma.During asthma, airways are persistently inflamed and sometimes may be constricted.

[0253] In some embodiments, the modified Cav-1 peptides disclosed herein are used to treat or prevent hypersensitivity interstitial pneumonitis in subjects. Hypersensitivity interstitial pneumonitis is a complex syndrome caused by inhalation of various antigens in susceptible and sensitized individuals. These antigens are found in the environment, mostly from avian proteins and fungi. Hypersensitivity interstitial pneumonitis is characterized by exaggerated humoral and cellular immune responses that affect small airways and lung parenchyma. Hypersensitivity interstitial pneumonitis can be classified into acute, chronic non-fibrotic, and chronic fibrotic forms. Acute hypersensitivity interstitial pneumonitis occurs as a result of intermittent high-level exposure to provoking antigens, usually within hours of exposure, whereas chronic hypersensitivity interstitial pneumonitis usually results from long-term low-level exposure (usually from birds or house mold) and is not easily defined in terms of time, and can occur within weeks, months, or even years of exposure. Some patients with fibrotic hypersensitivity interstitial pneumonia may develop a progressive phenotype even with complete avoidance of exposure. See Costabel et al., Nature Reviews Disease Primers 2020;6(65).

[0254] In some embodiments, modified Cav-1 peptides are used to treat or prevent systemic sclerosis or scleroderma in subjects. Systemic sclerosis is a systemic autoimmune disease characterized by endothelial dysfunction resulting in vasculopathy of small blood vessels, fibroblast dysfunction resulting in excessive collagen production and fibrosis, and immune abnormalities. The classification of systemic sclerosis is subdivided into diffuse cutaneous sclerosis, localized cutaneous sclerosis, or systemic scleroderma without cutaneous sclerosis based on the degree of skin involvement. Although virtually any organ system may be involved in the disease process, the fibrotic and vascular pulmonary manifestations of systemic sclerosis, including interstitial lung disease and pulmonary hypertension, are the major cause of death. Certain pulmonary manifestations may occur more commonly in subsets of systemic sclerosis (i.e., ILD is more common in diffuse cutaneous sclerosis, while pulmonary hypertension is more common in localized cutaneous sclerosis), but all known pulmonary manifestations reported have been described in each disease subset. Pulmonary disease can also occur in systemic sclerosis without skin involvement (known as scleroderma without scleroderma). See Solomon et al., Eur Respir Rev 2013;22(127):6-19.

[0255] In some embodiments, modified Cav-1 peptides are used to treat or prevent sarcoidosis in a subject. Sarcoidosis is a multisystem disorder characterized by non-caseating epithelioid cell granulomas that can affect almost any organ. Thoracic involvement is common and accounts for most of the morbidity and mortality associated with the disease. Thoracic abnormalities are observed in approximately 90% of sarcoidosis patients, and an estimated 20% develop chronic lung disease leading to pulmonary fibrosis. Pulmonary sarcoidosis can manifest in a variety of patterns. Bilateral hilar lymph node enlargement is the most common finding, followed by interstitial lung disease. The most typical findings of pulmonary involvement are micronodules with perilymphatic distribution, fibrosis, and bilateral perihilar opacities. Atypical manifestations such as mass-like or alveolar opacities, honeycomb cysts, miliary opacities, mosaic opacities, tracheobronchial involvement, and pleural disease, as well as complications such as aspergilloma, may also be seen. See Criado et al., Chest Imaging 2010;30(6).

[0256] In some embodiments, the present disclosure provides a method of treating or preventing a pulmonary disease or disorder in an elderly subject, the method comprising administering to the elderly subject an effective amount of a modified Cav-1 peptide or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of treating or preventing a pulmonary disease or disorder in an elderly subject, the method comprising administering to the elderly subject an effective amount of a modified Cav-1 peptide comprising an amino acid sequence of any one of SEQ ID NOs: 2-111. In some embodiments, the present disclosure provides a method of treating or preventing a pulmonary disease or disorder in an elderly subject, the method comprising administering to the elderly subject an effective amount of a modified Cav-1 peptide comprising an amino acid sequence of any one of SEQ ID NOs: 4-20. In some embodiments, the present disclosure provides a method of treating or preventing a pulmonary disease or disorder in an elderly subject, the method comprising administering to the elderly subject an effective amount of a modified Cav-1 peptide comprising an amino acid sequence of SEQ ID NO: 3. In some embodiments, the present disclosure provides a method of treating or preventing a pulmonary disease or disorder in an elderly subject, the method comprising administering to the elderly subject an effective amount of a modified Cav-1 peptide comprising an amino acid sequence of SEQ ID NO: 8. In some embodiments, the disclosure provides a method of treating or preventing a pulmonary disease or disorder in an elderly subject, the method comprising administering to the elderly subject an effective amount of a modified Cav-1 peptide comprising at least one amino acid substitution, insertion, deletion relative to the amino acid sequence FTTFTVT (SEQ ID NO: 3), wherein the modified Cav-1 peptide maintains the biological activity of Cav-1. In some embodiments, the method of administering the modified Cav-1 peptide further comprises nebulizing a solution comprising the modified Cav-1 peptide.

[0257] The present invention contemplates all modes of administration, doses, or dose frequencies suitable for treating or preventing the disease or disorder of interest. Effective doses can also be extrapolated from dose-response curves derived from in vitro or animal model test bioassays or systems.

[0258] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered intravenously, intrathecally, intradermally, transdermally, intrathecally, intraarterially, intraperitoneally, intranasally, intravaginally, intravesicularly, intraarticularly, intralesionally, intrarectally, intramuscularly, subcutaneously, mucosally, orally, topically, by inhalation (e.g., inhalation of aerosolized or dry powder formulations), by injection, by infusion, by continuous infusion, by localized perfusion that directly bathes target cells, via catheter, via lavage, in lipid compositions (e.g., liposomes), or by other methods known to those skilled in the art or any combination of the above (see, for example, Remington's Pharmaceutical Sciences, 18th Ed., 1990, which is incorporated herein by reference). The choice of injection volume and needle size can be selected by those skilled in the art based on the injection site, needle passability, and injectability, including considering the viscosity of the solution or suspension to be injected, as well as drug concentration, pH, and osmolality. In some cases, the particle size of the active agent can be selected to provide a desired dissolution rate upon administration (eg, by subcutaneous injection).

[0259] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered systemically. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered intravenously, intrathecally, subcutaneously, and / or intraperitoneally.

[0260] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is delivered locally to the subject's airway, such as administering a spray formulation using a nebulizer or a dry powder formulation using a dry powder inhaler. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the lungs of an elderly subject using a nebulizer. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the lungs of an elderly subject using a dry powder inhaler. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to an elderly subject intranasally, intrabronchially, intrapleurally, intratracheally, or via inhalation.

[0261] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject in a single dose or multiple doses. When multiple doses are administered, the doses can be separated from each other by, for example, one hour, three hours, six hours, eight hours, twelve hours, one day, two days, three days, four days, five days, six days, one week, two weeks, three weeks, one month, two months, three months, four months, five months, six months, one year, or any value or range therebetween. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered, for example, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every seven weeks, once every eight weeks, once every ten weeks, once every fifteen weeks, once every twenty weeks, or more. It will be understood that for any particular subject, the specific dosing regimen should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the composition. For example, the dosage of the modified Cav-1 peptide or pharmaceutical composition thereof can be increased if a lower dose does not provide sufficient therapeutic activity.

[0262] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 0.0001 mg / kg to about 1,000 mg / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 0.0001 mg / kg to about 0.01 mg / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 0.01 mg / kg to about 1 mg / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 1 mg / kg to about 100 mg / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 1 mg / kg to about 50 mg / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 1 mg / kg to about 25 mg / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 1 mg / kg to about 10 mg / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 10 mg / kg to about 25 mg / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 25 mg / kg to about 50 mg / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 50 mg / kg to about 75 mg / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 75 mg / kg to about 100 mg / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to a subject at a dose of about 0.0001 mg / kg, about 0.01 mg / kg, about 0.01 mg / kg, about 0.1 mg / kg, about 1 mg / kg, about 5 mg / kg, about 10 mg / kg, about 25 mg / kg, about 50 mg / kg, about 100 mg / kg, about 500 mg / kg, or about 1,000 mg / kg.

[0263] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 0.0001 g / kg to about 1,000 g / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 0.0001 g / kg to about 0.01 g / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 0.01 g / kg to about 1 g / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 1 g / kg to about 100 g / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 1 g / kg to about 50 g / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 1 g / kg to about 25 g / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 1 g / kg to about 10 g / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 10 mg / kg to about 25 mg / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 25 g / kg to about 50 g / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 50 g / kg to about 75 g / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered to the subject at a dose of about 75 g / kg to about 100 g / kg. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is provided at a dose of about 0.0001 g / kg, about 0.01 g / kg, about 0.01 g / kg, about 0.1 g / kg, about 1 g / kg, about 5 g / kg, about 10 g / kg, about 25 g / kg, about 50 g / kg, about 100 g / kg, about 500 g / kg, or about 1,000 g / kg.

[0264] In some embodiments, the total or complete dose of the modified Cav-1 peptide or pharmaceutical composition thereof administered to a subject is between about 1 mg and about 100 mg, e.g., between about 20 mg and about 100 mg, between about 50 mg and about 100 mg, between about 10 mg and about 20 mg, between about 20 mg and about 40 mg, between about 50 mg and about 70 mg, or between about 80 mg and about 90 mg.

[0265] In some embodiments, the dosage of modified Cav-1 peptide or pharmaceutical composition thereof for a particular subject is determined by those skilled in the art using conventional considerations (e.g., by suitable conventional pharmacological protocols). A physician may, for example, prescribe a relatively low dose initially and then increase the dose until an appropriate response is obtained. The dose administered to a subject is sufficient to provide a beneficial therapeutic response to the subject over time, or is sufficient to reduce symptoms or other appropriate activity, for example, depending on the application. The dosage is determined by the effectiveness of the particular formulation, as well as the activity, stability, and / or serum half-life of the modified Cav-1 peptide disclosed herein, as well as the condition of the subject, and the body weight or surface area of ​​the subject to be treated.

[0266] In some embodiments, a subject is administered a dose of modified Cav-1 peptide or pharmaceutical composition thereof once a day for the treatment or prevention of any one of the diseases or conditions described herein. In some embodiments, the subject is elderly. In some embodiments, a single dose is between about 0.2 mg / kg and about 250 mg / kg, e.g., between about 1 mg / kg and about 10 mg / kg, between about 10 mg / kg and about 25 mg / kg, between about 25 mg / kg and about 50 mg / kg, between about 50 mg / kg and about 75 mg / kg, between about 75 mg / kg and about 100 mg / kg, e.g., via pulmonary instillation (e.g., inhalation). Such doses can be administered daily, or at any frequency disclosed herein, ranging from about 3 days to a week or more. Long-term administration of modified Cav-1 peptide or pharmaceutical composition thereof is also possible, although the dose may need to be adjusted downward as is well understood in the art. However, the foregoing ranges are only suggestive because of the large number of variables in individual treatment regimens, and substantial deviations from these preferred values ​​are to be expected.

[0267] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered continuously to the subject. For continuous administration, for example by a pump system such as an osmotic pump, the total dosage over a period of about 1 to 2 weeks is preferably in the range of 1 mg / kg to 1 g / kg, preferably 20 to 300 mg / kg, more preferably 50 to 200 mg / kg. After such a continuous administration regimen, the total concentration of the active compound is preferably in the range of about 0.5 μM to about 50 μM, preferably about 1 μM to about 10 μM.

[0268] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered on a routine schedule. As used herein, a routine schedule refers to a predetermined, designated period of time. A routine schedule can encompass periods of the same or different lengths, so long as the schedule is predetermined. For example, a routine schedule can include administration once a day, twice a day, every other day, every third day, every fourth day, every fifth day, every sixth day, once a week, once every two weeks, once every three weeks, once a month, once every two months, once every three months, once every six months, or any set number of days, weeks, or months in between. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered on a twice-daily basis for the first week, followed by a once-daily basis for several months. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered once a day. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered less than once per day, such as every other day, every third day, or once a week.

[0269] In some embodiments, the modified Cav-1 peptide is administered to a subject with kidney disease or disorder for at least about 2 weeks, at least about 3 weeks, at least about 4 weeks, at least about 5 weeks, at least about 6 weeks, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 1 year, or any set number of weeks or months therebetween.In some embodiments, the modified Cav-1 peptide is administered to a subject with kidney disease or disorder for at least about 2 weeks.In some embodiments, the modified Cav-1 peptide is administered to a subject with kidney disease or disorder for at least about 4 weeks.

[0270] In some embodiments, the modified Cav-1 peptide is administered once a day to a subject with a kidney disease or disorder for at least about 2 weeks, at least about 3 weeks, at least about 4 weeks, at least about 5 weeks, at least about 6 weeks, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 1 year, or any set number of weeks or months therebetween. In some embodiments, the modified Cav-1 peptide is administered once a day to a subject with a kidney disease or disorder for at least about 2 weeks. In some embodiments, the modified Cav-1 peptide is administered once a day to a subject with a kidney disease or disorder for at least about 4 weeks.

[0271] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is provided in a unit dosage form (e.g., a pre-divided dose), such as a capsule, blister, or cartridge. In some embodiments, the unit dose comprises at least 1 mg of the modified Cav-1 peptide per dose, e.g., at least about 5 mg, at least about 10 mg, at least about 15 mg, or at least about 20 mg of the modified Cav-1 peptide. In some embodiments, the unit dose is about 1 mg to about 10 mg (e.g., about 5 mg) of the modified Cav-1 peptide. In some embodiments, the unit dosage form does not include the administration or addition of excipients and is merely used to hold the powder for inhalation (i.e., the capsule, blister, or cartridge is not administered). In some embodiments, two or more of the unit dosage forms are administered to the subject. For example, in the case of a dry powder inhaler, the modified Cav-1 peptide is provided in a unit dose capsule, and two or more unit dose capsules (e.g., 3-4) can be administered to the subject by inhalation. In some embodiments, the modified Cav-1 peptide is administered at a high emitted dose, such as at least about 10 mg, preferably at least about 15 mg, and even more preferably at least about 20 mg. In some embodiments, administration of the milled modified Cav-1 peptide results in a high fine particle dose in the deep lung, such as greater than about 5 mg. The fine particle dose in the deep lung is preferably at least about 10 mg, and even more preferably at least about 15 mg. In some embodiments, the fine particle dose is generated from 1, 2, 3, 4, or 5 or more capsules containing a dose of the peptide of the embodiment. In some embodiments, the fine particle dose is at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, or at least about 80% of the emitted dose.

[0272] In some embodiments, the change in inhalation pressure results in a change in the emitted dose. In some embodiments, a change in inhalation pressure of about 3 kPa, such as from about 4 kPa to about 1 kPa, results in a reduction in the emitted dose of less than about 25%, such as about 24%, about 23%, about 22%, about 21%, about 20%, about 19%, about 18%, about 17%, about 16%, about 15%, about 14%, about 13%, about 12%, about 11%, about 10%, about 9%, about 8%, about 7%, about 6%, about 5% or less. In some embodiments, the change in inhalation pressure results in a change in the fine particle dose. In some embodiments, a change in inhalation pressure of about 3 kPa, such as from about 4 kPa to about 1 kPa, results in a reduction in the fine particle dose of less than about 15%, such as about 14%, about 13%, about 12%, about 11%, about 10%, about 9%, about 8%, about 7%, about 6%, about 5% or less.

[0273] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered simultaneously or sequentially in combination with at least one additional therapeutic agent for the treatment or prevention of a kidney disease or disorder in a subject. In some embodiments, the disease is chronic kidney disease. The additional therapeutic agent may include, but is not limited to, angiotensin-converting enzyme (ACE) inhibitors, such as Capoten® (captopril), Vasotec® (enalapril), Monopril® (fasinopril), Prinivil® or Zestril® (lisinopril), or Altace® (ramipril); angiotensin II receptor (ARB) inhibitors, such as Edarbi® (azirsa), such as Teveten® (eprosartan), Avapro® (irbesartan), Cozaar® (losartan), Benicar® (olmesartan), or Diovan® (valsartan); Farxiga® (dapagliflozin); Aranesp® (darbepoetin alfa); and / or Procrit® or Epogen® (erythropoietin).

[0274] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered in combination with dialysis to treat kidney disease or disorder in a subject.In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered in combination with dialysis to treat chronic kidney disease.In some embodiments, the dialysis is hemodialysis.In some embodiments, the dialysis is peritoneal dialysis.

[0275] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered alone or in combination with at least one additional therapeutic agent in a subject. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered simultaneously or sequentially in combination with at least one additional therapeutic agent for the treatment or prevention of kidney disease or disorder in a subject. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered simultaneously or sequentially in combination with at least one additional therapeutic agent for the treatment or prevention of fibrotic disease or disorder in an elderly subject. The additional therapeutic agent includes, but is not limited to, nonsteroidal anti-inflammatory drugs (NSAIDs), steroids, disease-modifying antirheumatic drugs (DMARDs), immunosuppressants, biological response modifiers, bronchodilators, or antifibrotic agents such as pirfendon, agents whose antifibrotic mechanism of action is not fully understood but may involve TGF-beta blockade, nintedanib, broad tyrosine kinase blockers, or any other antifibrotic agent. Suitable NSAIDS are the non-selective cyclooxygenase (COX) inhibitors acetylsalicylic acid, mesalazine, ibuprofen, naproxen, flurbiprofen, fenoprofen, fenbufen, ketoprofen, indoprofen, pirprofen, carprofen, oxaprozin, pranoprofen, miroprofen, tioxaprofen, suprofen, alminoprofen, tiaprofenic acid, fluprofen, indomethacin, sulindac, tolmetin, zomepirac, nabumetone, diclofenac, and febufen. and pharmaceutically acceptable salts thereof, selected from anclofenac, alclofenac, bromfenac, ibufenac, aceclofenac, acemetacin, fentiazac, clidanac, etodolac, oxpinac, mefenamic acid, meclofenamic acid, flufenamic acid, niflumic acid, tolfenamic acid, diflunisal, flufenisal, piroxicam, tenoxicam, lonoxicam, and nimezuride, and the pharmaceutically acceptable salts thereof, the selective COX2 inhibitors meloxicam, celecoxib, and rofecoxib, and the pharmaceutically acceptable salts thereof.Suitable steroids are prednisone, prednisolone, methylprednisolone, dexamethasone, budenoside, fluocortolone, and triamcinolone.Suitable DMARDs are sulfasalazine, olsalazine, chloroquine, gold derivatives (auranofin), D-penicillamine, and cytostatics such as methotrexate and cyclophosphamide.Suitable immunosuppressants are cyclosporine A and its derivatives, mycophenolate mofetil, FK 506 (also known as tacrolimus and fujimycin), muromonab-CD3 (Orthoclone OKT-3®), antithymocyte globulin (ATG), 15-desoxyspergualin, mizoribine, misoprostol, rapamycin, leflunomide, and azathioprine. Suitable biological response modifiers are interferon beta, anti-TNF-α antibodies (etanercept), IL-10, anti-CD3 antibodies, or anti-CD25 antibodies. Suitable bronchodilators are ipratropium bromide, oxitropium bromide, tiotropium bromide, epinephrine hydrochloride, salbutamol, terbutaline sulfate, fenoterol hydrobromide, salmetreol, and formoterol. In such combinations, each active ingredient can be administered according to either its normal dosage range or a dose below its normal dosage range. The dosage of the combined NSAIDs, steroids, DMARDs, immunosuppressants, and biological response modifiers is suitably 1 / 50 of the normally recommended minimum dosage, up to 1 / 1 of the normally recommended dosage, preferably 1 / 20 to 1 / 2, more preferably 1 / 10 to 1 / 5. The usual recommended dosages of the combined drugs are to be understood as being, for example, the dosages disclosed in the Rote Liste® 2002, Editio Cantor Verlag Aulendorf, Germany, or in the Physician's Desk Reference.

[0276] In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered simultaneously or sequentially in combination with at least one additional therapeutic agent for treating or preventing kidney disease or disorder in a subject. In some embodiments, the modified Cav-1 peptide or pharmaceutical composition thereof is administered simultaneously or sequentially in combination with at least one additional therapeutic agent for treating pathogens or pathogen-induced lung injury in an elderly subject. The additional therapeutic agent includes, but is not limited to, chloroquine, hydroxychloroquine, type I interferon, antiviral agents, antibiotics, remdesivir, favipiravir, lopinavir, and ritonavir.

[0277] Hydroxychloroquine is a chemical derivative of chloroquine that features a hydroxyethyl group instead of an ethyl group. Hydroxychloroquine is classified as an effective antimalarial drug and has shown efficacy in the treatment of systemic lupus erythematosus as well as rheumatoid arthritis and Sjögren's syndrome. Hydroxychloroquine has been known for some time to increase lysosomal pH in antigen-presenting cells, but its mechanism of action in inflammatory conditions has only recently been elucidated and involves blocking the activation of Toll-like receptors on plasmacytoid dendritic cells. Hydroxychloroquine has shown efficacy in the treatment of RNA viruses, including hepatitis C. Hydroxychloroquine can be administered in doses of 600 mg per day.

[0278] Human type I interferons (IFNs) are a large subgroup of interferon proteins that help regulate the activity of the immune system. The mammalian types are designated IFN-α (alpha), IFN-β (beta), IFN-κ (kappa), IFN-δ (delta), IFN-ε (epsilon), IFN-τ (tau), IFN-ω (omega), and IFN-ζ (zeta, also known as limitin). Type I interferons have shown effectiveness against the replication of a variety of viruses, including Zika virus, Chikungunya virus, Flavivirus, and Hepatitis C virus. "Interferon compounds" include interferon-alpha, interferon-alpha analogs, interferon-alpha derivatives, interferon-alpha conjugates, interferon beta, interferon-beta analogs, interferon-beta derivatives, interferon-beta conjugates, and mixtures thereof. The entire protein or fragments thereof can be fused to other peptides and proteins, such as immunoglobulins and other cytokines. Interferon-alpha and interferon-beta conjugates can refer to compositions that include interferon-beta bound to a non-natural polymer, for example, a polyalkylene glycol moiety.Suitable interferon compounds include Roferon® (interferon alpha-2a), Intron® (interferon alpha-2b), Alferon® (interferon alpha-n3), Infergen® (interferon alphacon-1), Omniferon® (interferon alpha), interferon alphacon-1, interferon-alpha, interferon-alpha analogs, pegylated interferon-alpha, polymerized interferon-alpha, dimerized interferon-alpha, interferon-alpha conjugated to a carrier, interferon-alpha as an oral inhalant, interferon-alpha as an injectable composition, interferon-alpha as a topical composition, Roferon® (interferon alpha-2a) analogs, Intron® (interferon alpha-2b) analogs, Alferon® (interferon alpha-n3) analogs, and Infergen® ( Interferon alfacon-1 analogs, Omniferon® (interferon alpha) analogs, interferon alfacon-1 analogs, interferon beta, Avonex™ (interferon beta-1a), Betaseron™ (interferon beta-1b), Betaferon™ (interferon beta-1b), Rebif™ (interferon beta-1a), interferon-beta analogs, pegylated interferon-beta, polymeric interferon-beta , dimerized interferon-beta, interferon-beta conjugated to a carrier, interferon-beta as an oral inhalant, interferon-beta as an injectable composition, interferon-beta as a topical composition, Avonex™ (interferon beta-1a) analogs, Betaseron™ (interferon beta-1b) analogs, Betaferon™ (interferon beta-1b) analogs, and Rebif™ (interferon beta-1a) analogs.Alternatively, agents that induce the production of interferon-alpha or interferon-beta or mimic the action of interferon-alpha or interferon-beta may be employed. Interferon inducers include tilorone, poly(I)-poly(C), imiquimod, clidanimod, and bropirimine. EXAMPLES

[0279] Example 1: Efficacy of modified Cav-1 peptides in chronic kidney disease The aim of this study was to evaluate the efficacy of the modified Cav-1 peptide APi2355 in ameliorating renal fibrosis and disease in the Col4a3- / - mouse model of Alport syndrome. Alport syndrome mice (knockout) have reduced expression of Cav-1 in the glomerular tubules and develop glomerular fibrosis.

[0280] Study 1: Two weeks of treatment with saline or APi2355 in the Col4a3- / - mouse model of Alport syndrome

[0281] Because Col4a3- / - mice on the 129S1 / SvlmJ strain background exhibit rapid progression of end-stage renal disease by 11-13 weeks of age, this study evaluated the efficacy of APi2355 in chronic renal disease. 129S1 / SvlmJ Col4a3- / - develop end-stage renal disease at a faster rate compared to Col4a3- / - mice on a C57BL / 6 background. Five-week-old (cohort 2) and six-week-old (cohort 1) Col4a3- / - mice on the 129S1 / SvlmJ background were intraperitoneally injected daily with APi2355 (SEQ ID NO: 8, 0.0022 mg / g body weight) or saline for two weeks. Glomerular selective permeability was monitored by collecting urine before treatment and on the day of sacrifice and measuring the albumin to creatinine (A / C) ratio. Blood was collected on the day of sacrifice to monitor glomerular filtration rate by blood urea nitrogen (BUN) assay, and kidney tissue was harvested after sacrifice to assess renal injury and fibrosis. Mouse identification numbers were used to distinguish treatment groups (e.g., mouse identification numbers 1009, 1044, 1066, and 1069 for cohort 1 as shown in Figure 1A).

[0282] Figure 1A shows urinary protein gels from Cohort 1 and Cohort 2 Col4a3- / - mice before treatment with saline or APi2355 peptide, and Figure 1B shows urinary protein gels from Cohort 1 and Cohort 2 Col4a3- / - mice after daily intraperitoneal injections of APi2355 peptide or saline for two weeks. Figure 1C shows urinary albumin to creatinine ratios (g / mg) from Cohort 1 and Cohort 2 Col4a3- / - mice before and after two weeks of treatment with saline or APi2355 peptide.

[0283] FIG. 2 shows blood urea nitrogen levels in Col4a3− / − mice from cohort 1 and cohort 2 after treatment with saline or APi2355 peptide.

[0284] Figures 3A and 3B show immunofluorescence staining of collagen I and nidogen on kidney tissue from untreated Col4a3+ / - mice or from Col4a3- / - mice of cohort 1 after 2 weeks of treatment with saline or APi2355 peptide. Col4a3+ / - mice showed collagen I staining in the interstitium and Bowman's capsule, while saline and APi2355 treated Col4a3- / - mice showed additional staining in sclerotic glomeruli.

[0285] Figures 4A and 4B show immunofluorescence staining of collagen I and αSMA on kidney tissue from untreated Col4a3+ / − mice or from Col4a3− / − mice of cohort 1 after 2 weeks of treatment with saline or APi2355 peptide. Col4a3+ / − mice showed αSMA staining in interstitial vessels, and saline and APi2355 treated Col4a3− / − mice showed additional staining in sclerotic glomeruli.

[0286] Figures 5A and 5B show immunofluorescence staining of collagen I and nidogen on kidney tissue from untreated Col4a3+ / - mice or from Col4a3- / - mice of cohort 2 after 2 weeks of treatment with saline or APi2355 peptide. Col4a3+ / - mice showed collagen I staining in the interstitium and Bowman's capsule, while saline and APi2355 treated Col4a3- / - mice showed additional staining in sclerotic glomeruli.

[0287] Figures 6A and 6B show immunofluorescence staining of collagen I and αSMA on kidney tissue from untreated Col4a3+ / − mice or from Col4a3− / − mice of cohort 2 after 2 weeks of treatment with saline or APi2355 peptide. Col4a3+ / − mice showed αSMA staining in interstitial vessels, and saline and APi2355 treated Col4a3− / − mice showed additional staining in sclerotic glomeruli.

[0288] Overall, both saline- and APi2355-treated Col4a3− / − mice in cohort 1 and cohort 2 displayed severe sclerosis in the interstitium and many sclerotic glomeruli.

[0289] Study 2: Four weeks of treatment with saline or APi2355 in the Col4a3- / - mouse model of Alport syndrome

[0290] Additional studies were performed to evaluate the efficacy of modified Cav-1 peptides in ameliorating renal fibrosis and disease using the Col4a3− / − mouse model of Alport syndrome.

[0291] Four-week-old Col4a3- / - mice on a 129S1 / SvlmJ background were intraperitoneally injected daily with APi2355 (SEQ ID NO: 8, 0.0022 mg / g body weight) or saline for 4 weeks. Urine was collected before treatment at 4 weeks of age, and at 6, 7, and 8 weeks of age to monitor glomerular selective permeability by measuring albumin-to-creatinine (A / C) ratios. Blood was collected at 4 weeks of age before treatment and at 6 and 8 weeks of age after treatment to monitor glomerular filtration rate using a BUN assay, and kidney tissue was harvested after sacrifice to evaluate renal injury and renal fibrosis. Mouse identification numbers were used to distinguish treatment groups (e.g., mouse identification numbers 1248, 1267, and 1277-1282 as shown in Figure 7A).

[0292] Figure 7A shows gels of urinary proteins from 4-week-old Col4a3- / - mice before treatment with saline or APi2355 peptide. Figure 7B shows gels of urinary proteins from 6-week-old Col4a3- / - mice after daily intraperitoneal injections of APi2355 peptide or saline for 2 weeks, and Figure 7C shows gels of urinary proteins from 7- and 8-week-old Col4a3- / - mice after daily intraperitoneal injections of APi2355 peptide or saline for 3 or 4 weeks. Figure 7D shows the albumin to creatinine ratio (g / mg) in the urine of Col4a3- / - mice before treatment at 4 weeks of age and during treatment with saline or APi2355 peptide at 6, 7, and 8 weeks of age.

[0293] FIG. 8 shows blood urea nitrogen levels in Col4a3− / − mice before treatment at 4 weeks of age and during treatment with saline or APi2355 peptide at 6 and 8 weeks of age.

[0294] 9A to 9H show immunofluorescence staining of collagen I and nidogen on kidney tissue from untreated Col4a3+ / - mice or Col4a3- / - mice after 4 weeks of treatment with saline or APi2355 peptide.

[0295] Table A shows quantification of glomerular fibrosis in 8-week-old untreated Col4a3+ / - mice or Col4a3- / - mice after 4 weeks of treatment with saline or APi2355 peptide. [Table 4]

[0296] Figures 10A and 10B show immunofluorescence staining of caveolin-1 and laminin-111 on intestinal tissue from untreated Col4a3+ / - mice (left panel) or Col4a3- / - mice after 4 weeks of treatment with saline (center panel) or APi2355 peptide (right panel). Figures 10C to 10F show immunofluorescence staining of caveolin-1 and laminin-111 on kidney tissue from untreated Col4a3+ / - mice (left panel) or Col4a3- / - mice after 4 weeks of treatment with saline (center panel) or APi2355 peptide (right panel).

[0297] Figure 11A shows immunofluorescence staining of caveolin-1, collagen I, laminin-111, and nidogen on kidney tissue from untreated Col4a3+ / - and Col4a3- / - mice, or Col4a3- / - mice after 4 weeks of treatment with saline or APi2355 peptide. Figure 11B shows quantification of glomerular fibrosis in 8-week-old female Col4a3- / - mice after 4 weeks of treatment with saline or APi2355 peptide.

[0298] Overall, treatment of Col4a+ / - mice with APi2355 peptide starting at 4 weeks of age did not delay the progression of albuminuria and BUN but reduced the percentage of sclerotic glomeruli in females as assayed by collagen I deposition. These data indicate that APi2355 reduced glomerular fibrosis in a mouse model of Alport syndrome.

[0299] Example 2: Efficacy of modified Cav-1 peptides in an aged mouse model of pulmonary fibrosis The aim of this study was to evaluate the modified Cav-1 peptide CSP-7 in the bleomycin model of idiopathic pulmonary fibrosis and acute respiratory distress syndrome (ARDS). Aged male mice were selected to closely match the human population most vulnerable to ARDS.

[0300] Test 1

[0301] The efficacy of CSP-7 (SEQ ID NO: 3) was tested using aged male mice with a high senescent cell burden and therefore low regenerative capacity. Mice were between 74 and 76 weeks of age at the time of intratracheal bleomycin induction (4 U / kg body weight, n=5-8 mice / group). Total collagen in lung tissue was assessed using the Total Collagen Assay (QuickZyme Biosciences). Proteins in lung homogenates were first hydrolyzed, then hydroxyproline was oxidized and stained.

[0302] Figure 12 shows total collagen in lung homogenates of saline or bleomycin treated aged mice administered control peptide (CP-DPI) or CSP-7 (CSP7-DPI) via dry powder inhalation. Bleomycin treated mice administered CSP-7 showed a reduction in total collagen in the lungs compared to mice administered the control peptide.

[0303] Test 2

[0304] Bleomycin (BLM, 8 U / kg) was administered intratracheally to 73-74 week old mice (n=52), which was expected to result in approximately 50% of mice losing lungs due to BLM-induced lung injury. 14 days after instillation, CSP-7 and control peptides (CP) were administered to mice via dry powder inhalation (DPI) or intraperitoneal injection (IP) as shown in Table B below. 21 days after instillation, mice were sacrificed and blood and tissues were collected for analysis. Statistics were performed using Dunnett's multiple comparison test. [Table 5]

[0305] Figure 13 shows collagen in total lung homogenates of saline or BLM treated aged mice administered control peptide (CP-DPI) or CSP-7 (CSP7-DPI) via dry powder inhalation. BLM treated mice administered CSP-7 showed a decrease in total collagen in the lungs compared to mice administered the control peptide.

[0306] FIG. 14A shows total SMADs (tSMADs, top panel) and phosphorylated SMADs (pSMADs, bottom panel) in total lung homogenates of BLM-treated aged mice administered control peptide (CP-DPI) or CSP-7 (CSP7-DPI) by dry powder inhalation, or CSP-7 by intraperitoneal injection (IP).

[0307] FIG. 14B shows galectin-7 in total lung homogenates of BLM-treated aged mice administered control peptide (CP-DPI) or CSP-7 (CSP7-DPI) by dry powder inhalation, or CSP-7 by intraperitoneal injection (IP).

[0308] Example 3: Expression of human caveolin-1 in normal and fibrotic kidney tissues The aim of this study was to evaluate the expression of human caveolin-1 in renal tissues from normal subjects and subjects with fibrotic renal disease.

[0309] Renal tissues were collected from normal subjects and subjects with fibrotic kidney disease and then processed and immunostained for human caveolin-1.

[0310] As shown in Figure 15A, human caveolin-1 staining was detected in glomeruli of kidney tissues obtained from normal subjects. Specifically, human caveolin-1 was detected in parietal epithelial cells lining Bowman's capsule, as well as in endothelial cells of the inner capsule. As shown in Figure 15B, caveolin-1 expression was decreased in fibrotic glomeruli in subjects with chronic kidney disease compared to normal controls. Histology also showed significant endothelial destruction in addition to fibrotic glomeruli.

[0311] Numbered Embodiments of the Invention Without limiting the scope of the appended claims, the present disclosure describes the following numbered embodiments. Embodiment 1. 1. A method of treating or preventing a kidney disease or disorder in a subject, comprising: (a) consisting of any one of the amino acid sequences of SEQ ID NOs: 2 to 111; or (b) comprising any one of the amino acid sequences of SEQ ID NOs: 2 to 111; or (c) A method comprising administering to a subject an effective amount of a modified Cav-1 peptide comprising any one of the amino acid sequences of SEQ ID NOs: 2 to 111 having one or more amino acid substitutions, insertions, deletions, or chemical modifications. Embodiment 2. 2. The method of embodiment 1, wherein the modified Cav-1 peptide comprises L-amino acids. Embodiment 3. 2. The method of embodiment 1, wherein the modified Cav-1 peptide comprises D-amino acids. Embodiment 4. 2. The method of embodiment 1, wherein the modified Cav-1 peptide comprises both L- and D-amino acids. Embodiment 5. 5. The method according to any one of embodiments 1 to 4, wherein the modified Cav-1 peptide comprises deuterated residues. Embodiment 6. 6. The method of any one of embodiments 1 to 5, wherein the modified Cav-1 peptide comprises at least one non-standard amino acid. Embodiment 7. 7. The method of embodiment 6, wherein the non-standard amino acid is ornithine. Embodiment 8. 8. The method of any one of embodiments 1 to 7, wherein the modified Cav-1 peptide comprises an N-terminal modification. EMBODIMENT 9. 8. The method of any one of embodiments 1 to 7, wherein the modified Cav-1 peptide comprises a C-terminal modification. Embodiment 10. 8. The method according to any one of the preceding embodiments, wherein the modified Cav-1 peptide comprises an N-terminal modification and a C-terminal modification. Embodiment 11. 11. The method of embodiment 8 or 10, wherein the N-terminal modification is acylation. Embodiment 12. 11. The method of embodiment 9 or 10, wherein the C-terminal modification is amidation. Embodiment 13. 2. The method of embodiment 1, wherein the modified Cav-1 peptide comprises the amino acid sequence of FTTFTVT (SEQ ID NO:3). Embodiment 14. 2. The method of embodiment 1, wherein the modified Cav-1 peptide comprises the amino acid sequence of KASFTTFTVTKGS (SEQ ID NO: 4). EMBODIMENT 15. 2. The method of embodiment 1, wherein the modified Cav-1 peptide comprises the amino acid sequence KASFTTFTVTKGS-NH2 (SEQ ID NO:5). EMBODIMENT 16. 2. The method of embodiment 1, wherein the modified Cav-1 peptide comprises the amino acid sequence of aaEGKASFTTFTVTKGSaa (SEQ ID NO: 6). EMBODIMENT 17. 2. The method of embodiment 1, wherein the modified Cav-1 peptide comprises the amino acid sequence of aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 7). Embodiment 18. 2. The method of embodiment 1, wherein the modified Cav-1 peptide comprises the amino acid sequence Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8). EMBODIMENT 19. 2. The method of embodiment 1, wherein the modified Cav-1 peptide comprises the amino acid sequence of OASFTTFTVTOS (SEQ ID NO: 9). EMBODIMENT 20. 2. The method of embodiment 1, wherein the modified Cav-1 peptide comprises the amino acid sequence OASFTTFTVTOS-NH2 (SEQ ID NO: 10). EMBODIMENT 21. 21. The method of any one of embodiments 1 to 20, wherein the modified Cav-1 peptide comprises an internalization sequence. EMBODIMENT 22. 22. The method of embodiment 21, wherein the internalization sequence is located at the C-terminus of the peptide. EMBODIMENT 23. 22. The method of embodiment 21, wherein the internalization sequence is located at the N-terminus of the peptide. EMBODIMENT 24. 24. The method of any one of embodiments 1 to 23, wherein the modified Cav-1 peptide further comprises a cap at the N-terminus and / or C-terminus. 25. 25. The method of embodiment 24, wherein the modified Cav-1 peptide comprises a cap at the N-terminus and C-terminus. 26. 26. The method according to any one of the preceding embodiments, wherein the modified Cav-1 peptide is cyclized. EMBODIMENT 27. 27. The method according to any one of the preceding embodiments, wherein the modified Cav-1 peptide maintains the biological activity of native Cav-1 (SEQ ID NO: 1). 28. The method according to embodiment 1, wherein the modified Cav-1 peptide is a multimer comprising at least two peptides according to any one of embodiments 1 to 25. 29. 29. The method of embodiment 28, wherein a first peptide of the at least two peptides is essentially identical to a second peptide of the at least two peptides. EMBODIMENT 30. 29. The method of embodiment 28, wherein a first peptide of the at least two peptides is not identical to a second peptide of the at least two peptides. EMBODIMENT 31. 31. The method according to any one of embodiments 1 to 30, wherein the modified Cav-1 peptide is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, or orally. EMBODIMENT 32. 32. The method of embodiment 31, wherein the modified Cav-1 peptide is administered subcutaneously. EMBODIMENT 33. 32. The method of embodiment 31, wherein the modified Cav-1 peptide is administered intravenously. EMBODIMENT 34. The method according to any one of embodiments 1 to 33, wherein the kidney disease or disorder is selected from the group consisting of chronic kidney disease, end stage renal disease, glomerulonephritis, focal segmental glomerulosclerosis, renal fibrosis, polycystic kidney disease, IgA nephropathy, lupus nephritis, nephrotic syndrome, Alport syndrome, amyloidosis, Goodpasture's syndrome, granulomatosis with polyangiitis, or acute kidney injury. EMBODIMENT 35. 35. The method of embodiment 34, wherein the kidney disease or disorder is Alport Syndrome. EMBODIMENT 36. 34. The method of any one of the preceding embodiments, wherein the kidney disease or disorder is characterized by fibrosis. 37. The method according to any one of embodiments 1-36, wherein the method further comprises administering an effective amount of at least one additional therapeutic agent. 38. The method of embodiment 37, wherein the at least one additional therapeutic agent is an angiotensin-converting enzyme (ACE) inhibitor and / or an angiotensin II receptor (ARB) inhibitor. 39. The method according to any one of embodiments 1-38, wherein the method further comprises treating the subject with dialysis. EMBODIMENT 40. 40. The method according to any one of the preceding embodiments, wherein the modified Cav-1 peptide is administered as a composition comprising the modified Cav-1 peptide and at least one pharma- ceutically acceptable carrier or excipient. EMBODIMENT 41. 1. A method of treating or preventing a fibrotic disease or disorder in an elderly subject, comprising: (a) consisting of any one of the amino acid sequences of SEQ ID NOs: 2 to 111; or (b) comprising any one of the amino acid sequences of SEQ ID NOs: 2 to 111; or (c) A method comprising administering to a subject an effective amount of a modified Cav-1 peptide comprising any one of the amino acid sequences of SEQ ID NOs: 2 to 111 having one or more amino acid substitutions, insertions, deletions, or chemical modifications. EMBODIMENT 42. 42. The method of embodiment 41, wherein the modified Cav-1 peptide comprises the amino acid sequence of FTTFTVT (SEQ ID NO: 3). EMBODIMENT 43. 42. The method of embodiment 41, wherein the modified Cav-1 peptide comprises the amino acid sequence Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8). EMBODIMENT 44. The method of any one of embodiments 41-43, wherein the fibrotic disease or disorder is interstitial lung disease. EMBODIMENT 45. 45. The method of embodiment 44, wherein the interstitial lung disease is idiopathic pulmonary fibrosis. EMBODIMENT 46. 46. ​​The method according to any one of embodiments 41-45, wherein the modified Cav-1 peptide is administered to the lung. EMBODIMENT 47. 47. The method of embodiment 46, wherein the modified Cav-1 peptide is administered to the lungs via inhalation. 48. The method according to any one of embodiments 41 to 47, wherein the modified Cav-1 peptide is formulated for inhalation. 49. The method of embodiment 48, wherein the modified Cav-1 peptide is formulated for pressurized metered dose inhalation. EMBODIMENT 50. 49. The method of embodiment 48, wherein the modified Cav-1 peptide is formulated as a dry powder. EMBODIMENT 51. 51. The method of embodiment 50, wherein the dry powder comprising the modified Cav-1 peptide is essentially free of excipients. EMBODIMENT 52. 52. The method of embodiment 50 or 51, wherein the dry powder is produced by a spray drying process, air jet milling, ball milling, or wet milling. EMBODIMENT 53. The method of embodiment 48, wherein the modified Cav-1 peptide is formulated for nebulization. EMBODIMENT 54. The method of embodiment 47, wherein the modified Cav-1 peptide is administered to the subject using a nebulizer. EMBODIMENT 55. The method of embodiment 47, wherein the modified Cav-1 peptide is administered to the subject using an inhaler. EMBODIMENT 56. The method according to any one of embodiments 41-55, wherein the method further comprises administering to the subject a therapeutically effective amount of at least one additional therapeutic agent. EMBODIMENT 57. The method of embodiment 56, wherein the at least one additional therapeutic agent is chloroquine, hydroxychloroquine, remdesivir, favipiravir, lopinavir, or ritonavir. EMBODIMENT 58. The method of any one of embodiments 41-57, wherein the subject is at least 55 years old, at least 60 years old, at least 65 years old, at least 70 years old, at least 75 years old, at least 80 years old, at least 85 years old, or at least 90 years old.

[0312] Incorporation by Reference All references, articles, publications, patents, patent publications, and patent applications cited herein are incorporated by reference in their entirety for all purposes. However, mention of any references, articles, publications, patents, patent publications, and patent applications cited herein is not and should not be taken as an acknowledgment or any form of suggestion that they constitute pertinent prior art or form part of the common general knowledge in any country throughout the world.

Claims

1. 1. A method of treating or preventing a kidney disease or disorder in a subject, comprising: (a) consisting of any one of the amino acid sequences of SEQ ID NOs: 2 to 111; (b) comprising any one of the amino acid sequences of SEQ ID NOs: 2 to 111; or (c) A pharmaceutical composition comprising a modified Cav-1 peptide for use in a method comprising administering to said subject an effective amount of a modified Cav-1 peptide comprising any one of the amino acid sequences of SEQ ID NOs: 2 to 111 having one or more amino acid substitutions, insertions, deletions, or chemical modifications.

2. The modified Cav-1 peptide is (a) contains L-amino acids; (b) contains D-amino acids, or 2. The pharmaceutical composition of claim 1, comprising (c) an L-amino acid and a D-amino acid.

3. The pharmaceutical composition of claim 1 or 2, wherein the modified Cav-1 peptide comprises at least one non-standard amino acid.

4. 4. The pharmaceutical composition of claim 3, wherein the non-standard amino acid is ornithine.

5. The modified Cav-1 peptide is (a) contains an N-terminal modification; (b) comprises a C-terminal modification; or (c) The pharmaceutical composition of claim 1 or 2, comprising an N-terminal modification and a C-terminal modification.

6. The pharmaceutical composition of claim 5 , wherein the N-terminal modification is acylation or the C-terminal modification is amidation.

7. 3. The pharmaceutical composition of claim 1, wherein the modified Cav-1 peptide comprises the amino acid sequence FTTFTVT (SEQ ID NO: 3).

8. 3. The pharmaceutical composition of claim 1 or 2, wherein the modified Cav-1 peptide comprises the amino acid sequence of KASFTTFTVTKGS (SEQ ID NO: 4), aaEGKASFTTFTVTKGSaa (SEQ ID NO: 6), aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 7), OASFTTFTVTOS (SEQ ID NO: 9), or OASFTTFTVTOS-NH2 (SEQ ID NO: 10).

9. 3. The pharmaceutical composition of claim 1, wherein the modified Cav-1 peptide comprises the amino acid sequence Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8).

10. The pharmaceutical composition of claim 1 or 2, wherein the modified Cav-1 peptide comprises an internalization sequence.

11. The pharmaceutical composition of claim 10 , wherein the internalization sequence is located at the C-terminus of the peptide or the N-terminus of the peptide.

12. The modified Cav-1 peptide is (a) further comprising a cap at the N-terminus; (b) further comprising a cap at the C-terminus; or 3. The pharmaceutical composition of claim 1 or 2, further comprising (c) caps at the N-terminus and the C-terminus.

13. The pharmaceutical composition of claim 1 or 2, wherein the modified Cav-1 peptide is a multimer comprising at least two peptides of claim 1 or 2.

14. 3. The pharmaceutical composition of claim 1, wherein the modified Cav-1 peptide is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, or orally.

15. The pharmaceutical composition of claim 14, wherein the modified Cav-1 peptide is administered subcutaneously.

16. 3. The pharmaceutical composition of claim 1, wherein the kidney disease or disorder is selected from the group consisting of chronic kidney disease, end-stage renal disease, glomerulonephritis, focal segmental glomerulosclerosis, renal fibrosis, polycystic kidney disease, IgA nephropathy, lupus nephritis, nephrotic syndrome, Alport syndrome, amyloidosis, Goodpasture's syndrome, granulomatosis with polyangiitis, or acute kidney injury.

17. 17. The pharmaceutical composition of claim 16, wherein the kidney disease or disorder is Alport syndrome.

18. 3. The pharmaceutical composition of claim 1 or 2, wherein the kidney disease or disorder is characterized by fibrosis.

19. 3. The pharmaceutical composition of claim 1 or 2, wherein the method further comprises administering an effective amount of at least one additional therapeutic agent.

20. 20. The pharmaceutical composition of claim 19, wherein the at least one additional therapeutic agent is an angiotensin-converting enzyme (ACE) inhibitor and / or an angiotensin II receptor (ARB) inhibitor.

21. 3. The pharmaceutical composition of claim 1 or 2, wherein the method further comprises treating the subject with dialysis.

22. The pharmaceutical composition of claim 1 or 2, comprising the modified Cav-1 peptide and at least one pharmaceutically acceptable carrier or excipient.

23. 1. A method of treating or preventing a fibrotic disease or disorder in an elderly subject, comprising: (a) consisting of any one of the amino acid sequences of SEQ ID NOs: 2 to 111; (b) comprising any one of the amino acid sequences of SEQ ID NOs: 2 to 111; or (c) A pharmaceutical composition comprising a modified Cav-1 peptide for use in a method comprising administering to said subject an effective amount of a modified Cav-1 peptide comprising any one of the amino acid sequences of SEQ ID NOs: 2 to 111 having one or more amino acid substitutions, insertions, deletions, or chemical modifications.

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