Modified Cabolin-1 peptide preparation, method for producing the same, and method for using the same

Abstract

This specification provides a modified caveolin-1 (Cav-1) peptide and a pharmaceutical formulation, as well as a method of using the peptide or the formulation to treat a subject in need thereof. In particular, this specification provides a modified Cav-1 peptide formulation and a method of using the formulation to treat various diseases such as fibrosis by providing long-term release of the peptide over a desired period of time.

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of priority of U.S. Provisional Application No. 63 / 368,169, filed on July 12, 2022, the contents of which are incorporated herein by reference in their entirety.

[0002] The present disclosure relates to a long - acting release formulation comprising a modified caveolin - 1 peptide. The present disclosure also provides methods of using such formulations for the treatment of various diseases including, but not limited to, fibrosis.

[0003] Incorporation of Sequence Listing The sequence listing associated with this application is provided in extensible markup language (XML) format instead of paper and is incorporated herein by reference. A computer - readable copy of the sequence listing (filename: LUTX_025_01WO_SeqList_ST26, creation date: July 6, 2023, file size: 162,438 bytes) has been electronically submitted.

Background Art

[0004] Caveolin-1 (Cav-1) is an endogenous membrane protein that has a homeostatic function in the fibrotic process by participating in a series of major regulatory pathways such as TGF-β signaling (Shihata et al., Front. Pharmacol. 2017, 8:567, and Gvaramia et al., Matrix Biol, 2013:32(6):307-315). Endogenous Caveolin-1 is constitutively suppressed in fibrotic lungs in IPF and in bleomycin-induced animal models of IPF patients (Wang et al., J Exp Med, 2006, 203(13):2895-2906, Sanders et al., PLoS One, 2015, 10(2), e0116995, and Sanders et al., Am J Respir Cell Mol Biol, 2017, 56(1):50-61). The full-length Caveolin-1 protein may be a less desirable pharmaceutical candidate due to concerns inherent in protein drugs, including stability, delivery, cost, and autoimmunogenicity. Therefore, fragments of Caveolin-1, such as Caveolin scaffold domain peptides (CSPs), have been studied and have been found to be effective alternatives to the full-length Caveolin-1 protein. In particular, the 20mer form of CSP has been shown to prevent, limit, or reverse fibrosis in animal models, and a 7-amino acid fragment of CSP, named CSP-7, was sufficiently effective at promoting fibrosis reduction both in vitro and in vivo (Marudamuthu et al., Sci Transl Med, 2019, 11(522), eaat2848). As a result, CSP has been identified as a promising therapeutic agent for the treatment of fibrotic diseases such as IPF.

[0005] Peptide therapeutics generally have a short half-life in vivo and require frequent administration of the therapeutic agent. Therefore, there is a need for long-release formulations for modified Cav-1 peptide therapeutics that reduce the burden of frequent administration and, in addition, minimize the adverse effects caused by fluctuations between the maximum and minimum serum drug concentrations. SUMMARY OF THE INVENTION

[0006] The present disclosure provides a formulation comprising a modified caveolin-1 peptide and at least one pharmaceutically acceptable carrier or excipient.

[0007] In one aspect, the formulation comprises a polypeptide comprising an amino acid sequence having a core sequence of FTTFTVT and sucrose acetate isobutyrate (SAIB).

[0008] In some embodiments, the formulation comprising SAIB further comprises a first solvent selected from N-methylpyrrolidone (NMP), absolute ethanol, or ethyl acetate. In some embodiments, the first solvent is NMP or ethyl acetate. In some embodiments, SAIB is present in the first solvent at about 50% w / w to about 95% w / w or 70% w / w to about 90% w / w. In some embodiments, SAIB is present in the first solvent at about 80% w / w.

[0009] In some embodiments of the formulation comprising SAIB, the polypeptide is micronized. In some embodiments, the average particle size of the polypeptide ranges from about 0.5 μm to about 100 μm. In some embodiments, the average particle size of the polypeptide ranges from about 1 μm to about 5 μm.

[0010] In some embodiments, the formulation comprising SAIB is in the form of an emulsion, solution, microsphere, nanoparticle, nanosphere, implant, or gel. In some embodiments, the formulation is a suspension.

[0011] In some embodiments, the formulation comprising SAIB further comprises a second solvent. In some embodiments, the second solvent is sterile water, phosphate buffered saline (PBS), or any pharmaceutically acceptable carrier.

[0012] In some embodiments, the formulation comprises the polypeptide and SAIB in a weight ratio of about 1:50 to about 1:1. In some embodiments, the weight ratio is about 1:30 to about 1:1.

[0013] In some embodiments, the formulation comprising the polypeptide and SAIB releases at least about 1% to about 30% of the polypeptide within 4 hours under physiological conditions. In some embodiments, the formulation releases at least about 5% to 25% of the polypeptide within 4 hours under physiological conditions. In some embodiments, the formulation comprising the polypeptide and SAIB releases at least about 1% to about 30% or at least about 5% to 25% of the polypeptide within 4 hours when measured in an aqueous buffer in the range of about pH 6 to about pH 8 at about 37°C. In some embodiments, the formulation comprising the polypeptide and SAIB releases at least about 1% to about 30% or at least about 5% to about 25% of the polypeptide within 4 hours when measured in an aqueous buffer at about pH 7.4 at about 37°C.

[0014] In some embodiments of any one of the SAIB formulations described herein, the polypeptide comprises ASFTTFTVTK. In some embodiments, the polypeptide consists of 20 or fewer amino acids. In some embodiments, the polypeptide comprises at least one amino acid added to the N-terminus, at least one amino acid added to the C-terminus, or at least one amino acid added to both the N-terminus and the C-terminus. In these embodiments, the addition made within 5 amino acids of each terminus has an amino acid sequence that is less than 80% identical to the contiguous amino acid sequence of SEQ ID NO: 1 or 2 (human Cav-1 peptide). In some embodiments, the addition made within 5 amino acids of each terminus has an amino acid sequence that is less than 60% identical to the contiguous amino acid sequence of SEQ ID NO: 1 or 2. In some, the addition made within 5 amino acids of each terminus has an amino acid sequence that is less than 40% identical to the contiguous amino acid sequence of SEQ ID NO: 1 or 2. In some embodiments, the addition made within 5 amino acids of each terminus has an amino acid sequence that is less than 20% identical to the contiguous amino acid sequence of SEQ ID NO: 1 or 2. In some embodiments, the polypeptide sequence includes the peptide containing L-amino acids, the peptide containing D-amino acids, or the peptide containing both L-amino acids and D-amino acids.

[0015] In some embodiments, the polypeptide comprises at least one non-standard amino acid, or the polypeptide comprises two non-standard amino acids. In some embodiments, the non-standard amino acid is ornithine. In some embodiments, the polypeptide further comprises an N-terminal modification, a C-terminal modification, or both an N-terminal and a C-terminal modification. In some embodiments, the N-terminal modification is acylation and / or the C-terminal modification is amidation.

[0016] In some embodiments of any one of the SAIB formulations described herein, the polypeptide comprises the amino acid sequence of FTTFTVT (SEQ ID NO: 3), KASFTTFTVTKGS (SEQ ID NO: 4), KASFTTFTVTKGS-NH2 (SEQ ID NO: 5), aaEGKASFTTFTVTKGSaa (SEQ ID NO: 6), aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 7), Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8), OASFTTFTVTOS (SEQ ID NO: 9), or OASFTTFTVTOS-NH2 (SEQ ID NO: 10). In some embodiments, the polypeptide consists of an amino acid sequence selected from FTTFTVT (SEQ ID NO: 3), KASFTTFTVTKGS (SEQ ID NO: 4), KASFTTFTVTKGS-NH2 (SEQ ID NO: 5), aaEGKASFTTFTVTKGSaa (SEQ ID NO: 6), aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 7), Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8), OASFTTFTVTOS (SEQ ID NO: 9), or OASFTTFTVTOS-NH2 (SEQ ID NO: 10). In some embodiments, the polypeptide consists of the amino acid sequence of Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8).

[0017] In yet another aspect, the present disclosure provides a method of practicing it in a subject in need of treatment of a disease, the method comprising administering to the subject a pharmaceutically effective amount of any one of the formulations described herein. In some embodiments, the disease is fibrosis. In some embodiments, the fibrosis is interstitial lung disease, liver fibrosis, kidney fibrosis, skin fibrosis, glomerulonephritis, systemic sclerosis, cardiac fibrosis, myocardial fibrosis, renal fibrosis, cirrhosis, nephrosclerosis, atherosclerosis, macular degeneration, ocular scarring, cataract, retinopathy and vitreoretinopathy, Graves ophthalmopathy, neurofibromatosis, scleroderma, glioblastoma, keloid and hypertrophic scarring, peritoneal fibrotic disease, chronic obstructive pulmonary disease, postoperative myoma, diabetic nephropathy, gynecological cancer, myeloproliferative syndrome, myeloid leukemia, myelodysplastic syndrome, inflammatory bowel disease, non-alcoholic fatty liver disease, fibrosarcoma, rheumatoid arthritis, non-alcoholic steatohepatitis, Alport syndrome, or chronic COVID syndrome.

[0018] In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis, familial pulmonary fibrosis, idiopathic nonspecific interstitial pneumonia, usual interstitial pneumonia, idiopathic organizing pneumonia, or sarcoidosis. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0019] In some embodiments, the disease is a renal disease or renal disorder. In some embodiments, the renal disease or renal disorder is selected from the group consisting of chronic kidney disease, end-stage renal disease, glomerulonephritis, focal segmental glomerulosclerosis, kidney 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 renal disease or renal disorder is kidney fibrosis. In some embodiments, the renal disease or renal disorder is Alport syndrome. In some embodiments, the administration is intravitreal, intradermal, transdermal, intramuscular, or subcutaneous administration. In some embodiments, the subject is a human.

[0020] In some embodiments of any one of the formulations described herein, the formulation releases the peptide for at least 7 days after a single-dose administration. In some embodiments, the formulation releases the polypeptide for at least 21 days after a single-dose administration. In some embodiments, the formulation releases the polypeptide for at least 28 days after a single-dose administration. In some embodiments, the formulation is administered to a subject at a dose of about 0.01 mg / kg to about 250 mg / kg. In some embodiments, the formulation is administered to a subject at a dose of about 0.05 mg / kg to about 50 mg / kg.

[0021] It is contemplated that any method or formulation described herein can be practiced with respect to any other method or formulation described herein. Other objects, features, and advantages of the present disclosure will become apparent from the following detailed description. However, while this detailed description shows certain embodiments of the present disclosure, it is to be understood that this detailed description is given by way of illustration only, since various changes and modifications within the spirit and scope of the present disclosure will become apparent to those skilled in the art from this detailed description.

Brief Description of the Drawings

[0022]

Figure 1

Figure 2

Modes for Carrying Out the Invention

[0023] The present disclosure overcomes the problems associated with the current art by providing modified caveolin-1 (Cav-1) peptides in a drug-controlled release dosage form. In some embodiments, pharmaceutical formulations of the modified Cav-1 peptides are provided. In some embodiments, the peptides are formulated for subcutaneous delivery. In some embodiments, the peptides are formulated for long-term release. Also provided herein is a method of treating or preventing fibrosis or chronic kidney disease by administering to a subject a therapeutically effective amount of any one of the modified Cav-1 peptide formulations described herein.

[0024] I. Definitions As used herein, the articles “a” or “an” refer to one or more than one (i.e., at least one) of the grammatical object of the article. As used herein, in the claims (s), when used in conjunction with the term “comprising”, the articles “a” or “an” refer to one or more than one of the grammatical object of the article.

[0025] The use of the term “or” in the claims is used to mean “and / or” unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive, even though the present disclosure supports a definition that the disclosure refers to only alternatives and “and / or”. As used herein, “another” may mean at least a second or later.

[0026] As used herein, the term “about” indicates that a value includes a value that is within 10% of the determined value, the inherent error variation of the device, method used to determine that value, the variation existing between test subjects, or the defined value.

[0027] The term "peptide" or "polypeptide" typically refers to an amino acid sequence composed of a single chain of amino acids linked by peptide bonds. In its broadest sense, this term can be used interchangeably with "protein" 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 such as, for example, esters, ethers, etc. As used herein, the term "amino acid" refers to any natural and / or non-natural or synthetic amino acid, including glycine and both D and L optical isomers, as well as amino acid analogs and peptidomimetics.

[0028] Generally, a peptide or polypeptide contains at least two amino acid residues and has a length of less than about 50 amino acids (e.g., 40 amino acids, 30 amino acids, 20 amino acids, or any number therein) unless otherwise specified. In some embodiments, a counterion is provided for the peptide or polypeptide. In some embodiments, the peptide or polypeptide includes N-terminal and / or C-terminal modifications such as blocking modifications that reduce degradation.

[0029] The term "peptidomimetic" or "peptide mimic" means that a peptide according to the present invention is 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. When the peptide chain is short, a peptide of three or more amino acids is generally called an oligopeptide. When the peptide chain is long, the peptide is generally called a peptide or a protein.

[0030] In some embodiments, a peptide or polypeptide has a certain percentage (e.g., 80%, 85%, 90%, or 95%) of "sequence identity" or "homology" to another sequence, which means that when aligned, that percentage of amino acids in the two sequences is the same. 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 corresponding residues of the sequence being compared, after the sequences are aligned, gaps are introduced if necessary to obtain the maximum percentage of identity over the entire sequence, and without considering any conservative substitutions as part of sequence identity. In some embodiments, N-terminal or C-terminal extensions or insertions are not interpreted as a decrease in identity or homology. Instead, in some embodiments, an N-terminal or C-terminal extension or insertion results in the newly formed sequence being non-homologous to the original sequence. Alignment and the percentage of homology or sequence identity can be determined using software programs known in the art, such as the programs described in Current Protocols In Molecular Biology (F.M. Ausubel et al., eds., 1987) Supplement 30, Section 7.7.18, Table 7.7.1.

[0031] The terms "insertion" or "deletion" typically range from about 1 to 5 amino acids. Permissible variations can be determined experimentally by synthetically generating the peptide while systematically making nucleotide insertions, deletions, or substitutions within the sequence using recombinant DNA techniques.

[0032] The term "substitution" when referring to a peptide refers to the change of one 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 has been replaced with a different natural or non-conventional amino acid residue. Such substitutions can be classified as "conservative", in which case the amino acid residue contained in the peptide has been replaced with another natural amino acid having similar characteristics with respect to either 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", in which case the amino acid residue present in the peptide has been replaced with an amino acid having 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 has been replaced with a non-conventional amino acid. In some embodiments, the amino acid substitution is conservative. In some embodiments, the amino acid substitution is non-conservative.

[0033] An "analogue" of a molecule such as a peptide refers to a molecule whose function is similar to either the whole molecule or a fragment thereof. The term "analogue" is also intended to include allelic species and induced variants. Analogues typically differ from the native peptide at one or a few positions, often by conservative substitutions. Analogs typically exhibit at least 80 or 90% sequence identity with the native peptide. Some analogues also include non-natural amino acids or modifications of the N- or C-terminal amino acids. Examples of non-natural amino acids include, but are not limited to, disubstituted amino acids, N-alkyl amino acids, lactic acid, 4-hydroxyproline, γ-carboxyglutamic acid, ε-N,N,N-trimethyllysine, ε-N-acetyllysine, O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylysine, and σ-N-methylarginine. Fragments and analogues can be screened for prophylactic or therapeutic efficacy in transgenic animal models as described below.

[0034] The term "covalently linked" refers to a peptide or polypeptide linked either directly or indirectly (e.g., via a linker) by a covalent chemical bond. In some embodiments, the fusion peptide is covalently bonded.

[0035] As used herein, the term "fusion protein" refers to a recombinant protein of two or more proteins. A fusion protein can be generated, for example, by constructing a single open reading frame in which a nucleic acid sequence encoding one protein is linked to a nucleic acid encoding another protein such that they are translated into a single peptide carrying all the proteins intended in the cell. The order of protein placement can vary. The fusion protein can include an epitope tag or a half-life extender. Examples of 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. Examples of half-life extenders include the Fc domain and serum albumin.

[0036] A "biologically active" caveolin-1 (Cav-1) peptide refers to a polypeptide that increases p53 protein levels, decreases urokinase plasminogen activator (uPA) and uPA receptor (uPAR), and / or increases plasminogen activator inhibitor-1 (PAI-1) expression in cells such as fibrotic lung fibroblasts. In some embodiments, the biologically active peptide has at least 20% of the biological or biochemical activity of the native Cav-1 peptide of SEQ ID NO: 1 (e.g., as measured by in vitro or in vivo assays). In some embodiments, the biologically active peptide has increased biological or biochemical activity compared to the native Cav-1 peptide.

[0037] As used herein, the term "isolated" refers to a peptide or polypeptide that has been separated from any natural environment, such as a body fluid like blood, and separated from the components that are naturally associated with the peptide.

[0038] As used herein, the term "essentially free of" with respect to a recited component means that none of the recited components are intentionally formulated in the formulation or composition herein and / or are present only as an impurity or in trace amounts. Thus, the total amount of the recited component resulting from any unintended impurity of the formulation is well below 0.01%. Most preferably, the formulation is such that no amount of the recited component can be detected using standard analytical methods.

[0039] The term "substantially pure" refers to a peptide or polypeptide that has been isolated and purified to some extent from the components that are naturally associated with it. Typically, a peptide or polypeptide is substantially pure if it contains 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% less of the proteins and naturally occurring organic molecules to which it is naturally associated. For example, a substantially pure peptide or polypeptide may be obtained by extraction from a natural source, typically by expression of a recombinant nucleic acid in a cell that does not normally express the protein, or by chemical synthesis.

[0040] The terms "subject", "individual", and "patient" are used interchangeably herein and refer to an animal, e.g., a human or non-human animal (e.g., a mammal), to which treatment, including prophylactic treatment, with a modified Cav-1 peptide or a pharmaceutical formulation thereof as disclosed herein is provided. As used herein, the term "subject" refers to human and non-human animals. The term "non-human animal" includes all vertebrates, e.g., non-human primates (especially higher primates), sheep, dogs, rodents (e.g., mice or rats), guinea pigs, goats, pigs, cats, rabbits, cows, etc. mammals, and non-mammals such as chickens, amphibians, reptiles, etc. In some embodiments, the subject is a human. In some embodiments, the subject is an experimental animal or animal substitute 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 aspects, the non-human animal is a companion animal such as a dog or a cat.

[0041] "Treating" a disease or condition in a subject or "treating" a patient having a disease or condition refers to administering a pharmaceutical treatment, e.g., administering a drug, to an individual such that at least one symptom of the disease or condition is reduced or stabilized. Typically, when a peptide of the present disclosure is administered therapeutically as a treatment, it is administered to a subject presenting with one or more symptoms of pathogen-mediated lung injury.

[0042] As used herein, the term "variant" refers to a peptide or nucleic acid that differs from the peptide 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 a naturally occurring molecule. Amino acid substitutions include changes in which an amino acid has been replaced with a different native or non-conventional amino acid residue. Such substitutions can be classified as "conservative", in which case the amino acid residue included in the peptide has been replaced with another native amino acid having similar characteristics with respect to either 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", in which case the amino acid residue present in the peptide has been replaced with an amino acid having different properties, such as a native amino acid from a different group (e.g., replacing a charged or hydrophobic amino acid with alanine), or a native amino acid has been replaced with a non-conventional amino acid. In some embodiments, the amino acid substitutions are conservative. Also included as variants when used with respect to a polynucleotide or peptide are polynucleotides or peptides that may have a different primary, secondary, or tertiary structure as compared to a reference polynucleotide or polypeptide (e.g., as compared to a wild-type polynucleotide or polypeptide).

[0043] The term "micronized" refers to a substance having a size of several microns.

[0044] The phrases "effective amount" or "therapeutically effective amount" mean a dosage of a drug or agent sufficient to produce a desired therapeutic result. The desired therapeutic result can be subjective or objective improvement in the recipient of the dosage, reduction of infection, reduction of inflammation, increase in lung growth, increase in lung repair, reduction of tissue edema, increase in DNA repair, decrease in apoptosis, decrease in tumor size, decrease in the rate of cancer cell proliferation, decrease in metastasis, or any combination of the above.

[0045] As used herein, "excipient" refers to a pharmaceutically acceptable carrier, which is a relatively inert substance used to facilitate the administration or delivery of an active pharmaceutical ingredient (API) to a subject, or to facilitate the processing of an API into a pharmaceutical dosage form that can be used as a medicine for delivery to a site of action within the subject. An excipient or pharmaceutically acceptable carrier includes all of the inert components of a dosage form, excluding the active ingredient(s). Non-limiting examples of excipients include carriers, diluents, stabilizers, surfactants, surface modifiers, solubilizers, buffers, encapsulating agents, antioxidants, preservatives, non-ionic wetting or clarifying agents, thickeners, and absorption enhancers. "Excipient-free" refers to a modified Cav-1 peptide pharmaceutical formulation that does not contain any excipients.

[0046] The term "physiological conditions" refers to a set of conditions including temperature, salt concentration, and pH that mimic the state of a living subject. This includes physiologically relevant conditions for use in in vitro assays. Generally, physiological buffers contain salts at physiological concentrations and are adjusted to a neutral pH in the range of about 6 to about 8, about 6.5 to about 7.8, or about 7.0 to about 7.5. Physiologically relevant temperatures are in the range of about 25°C to about 38°C, or about 30°C to about 37°C. In some embodiments, the physiological buffer is a phosphate buffer, such as a phosphate buffer or phosphate buffered saline at about 10 to about 100 mM, or a simulated body fluid. In some embodiments, the physiological buffer is made isotonic (e.g., about 250 to about 350 mOsm / kg) with a soluble material including but not limited to sodium chloride or dextrose. In some embodiments, the physiological buffer may also include a surfactant.

[0047] The terms "pharmaceutical formulation", "pharmaceutically acceptable formulation", or "pharmacologically acceptable formulation" refer to molecular entities and formulations that do not, when administered to animals such as humans as necessary, cause adverse reactions, allergic reactions, or other undesirable reactions. The preparation of pharmaceutical formulations containing modified Cav-1 peptides such as CSP7 or additional active ingredients will be known to those skilled in the art in light of the present disclosure. Further, for administration to animals (e.g., humans), it will be understood that the preparation should meet the standards of bioburden, sterility, pyrogenicity, general safety, and / or purity required by the FDA or other recognized regulatory authorities.

[0048] As used herein, "pharmaceutically acceptable carrier" includes every excipient, processing aid, aqueous solvent (e.g., water, alcoholic / aqueous solutions, saline solutions, 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), dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial or antifungal agents, antioxidants, chelating agents, and inert gases), isotonic agents, absorption delaying agents, salts, drugs, drug stabilizers, gels, binders, excipients, disintegrants, lubricants, flavoring agents (e.g., sweetening agents, flavoring agents), similar materials known to those skilled in the art, and combinations thereof. The pH and exact concentration of the various components in a pharmaceutical formulation are adjusted according to well-known parameters. In some embodiments, the carrier may encapsulate a therapeutic agent but is not itself consumed or administered to the subject (e.g., a shell capsule enclosing a dry powder formulation for use in a dry powder inhaler). See, e.g., Remington’s Pharmaceutical Sciences, 18th Ed., 1990, which is incorporated herein by reference.

[0049] II. Caveolin-1 Peptide Embodiments of the present disclosure provide modified versions of the natural caveolin-1 (Cav-1) protein, including but not limited to fragments, derivatives, and variants of the natural Cav-1 protein. In some embodiments, the modified Cav-1 peptide is a truncated form of a natural Cav-1 peptide, such as the exemplary peptides shown in Table 2 and / or Table 3.

[0050] Natural 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.

[0051]

Table 1

[0052] 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 essential role in the dimerization of caveolin-1 and the regulation of 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 the activation of Wnt signaling, β-catenin-mediated transcription, 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 the interaction of Cav-1 with SRC kinase and mimics the combined effect of uPA and anti-β1-integrin antibody. The endogenous CSD domain can form homodimers with other Cav-1 proteins and interact with proteins having a caveolin-binding domain sequence (CBD) motif.Up to 30% of all endogenous proteins are estimated to have a CBD motif, and the CSD domain of caveolin-1 is hypothesized to provide stability to these proteins (see Marudamuthu et al., Am J Pathol 2015, 185:55-68). Treatment with CSP 20mer (the full-length CSD of caveolin-1) resulted in decreased lung αSMA and lung epithelial apoptosis, decreased collagen deposition, and downregulation of the expression of pro-fibrotic signaling molecules (see Bhandary et al., Am J Phys Lung Cell Mol Phys, 2012, 302(5), L463-L473, Razani et al., JBC, 2001, 276(9), 6727-6738, and Lee et al., Biochem Biophys Res Commun, 2007, 359(2):385-390).

[0053] In some embodiments, the modified Cav-1 peptide is CSP-7. CSP-7 is a 7-amino acid fragment of the human CSD of caveolin-1 (see SEQ ID NO: 3 in Table 3 below).

[0054] Exemplary amino acid sequences of the modified Cav-1 peptides are shown in Tables 2 and 3 below. Capital letters mean L-amino acids and lowercase letters mean D-amino acids (for example, lowercase "a" represents D-alanine). The term "Ac" refers to an acetyl group and the term "NH2" refers to an amide group. "O" means ornithine.

[0055]

Table 2

[0056]

Table 3-1

Table 3-2

[0057] 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, b) containing the core sequence of any one of the amino acid sequences of SEQ ID NOs: 2 to 111, or c) containing the core sequence of any one of the amino acid sequences of SEQ ID NOs: 2 to 111, wherein the core sequence comprises one or more amino acid substitutions, insertions, deletions, or chemical modifications.

[0058] In some embodiments, the Cav-1 peptide comprises or consists of the amino acid sequence of SEQ ID NO: 1. In some embodiments, the Cav-1 peptide has 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 and comprises an amino acid sequence. In some embodiments, the Cav-1 peptide comprises the amino acid sequence of SEQ ID NO: 1 and has one or more mutations relative thereto. For example, in some embodiments, the Cav-1 peptide 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 peptide comprises the amino acid sequence of SEQ ID NO: 1 and has 1 to 5, 5 to 10, 11 to 5, 15 to 20, 10 to 25, 25 to 30, or more than 30 mutations. In some embodiments, the modified Cav-1 peptide comprises 1, 2, 3, 4, 5, or more amino acid substitutions, deletions, or insertions relative to the sequence of SEQ ID NO: 1 that result in a peptide of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 residues.

[0059] In some embodiments, the modified Cav-1 peptide comprises, or consists of, an amino acid sequence of any one of SEQ ID NOs: 2 to 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 to 111. In some embodiments, the modified Cav-1 peptide comprises an amino acid sequence of any one of SEQ ID NOs: 2 to 111 and has one or more mutations relative 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 any one of SEQ ID NOs: 2 to 111. In some embodiments, the modified Cav-1 peptide comprises an amino acid sequence of any one of SEQ ID NOs: 2 to 111 and has 1 to 5, 5 to 10, or 11 to 15, or more mutations. In some embodiments, the modified Cav-1 peptide comprises 1 to 5 additional amino acids at either or both of the N-terminus and / or C-terminus of any one of SEQ ID NOs: 2 to 111.

[0060] In some embodiments, the modified Cav-1 peptide comprises or consists of the amino acid sequences of SEQ ID NOs: 2-10. 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 NOs: 2-10. In some embodiments, the modified Cav-1 peptide comprises the amino acid sequence of SEQ ID NO: 3 and has 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 NOs: 2-10. In some embodiments, the modified Cav-1 peptide comprises an additional 1-5 amino acids at either or both of the N-terminus and / or C-terminus of SEQ ID NOs: 2-10. In some embodiments, the modified Cav-1 peptide of SEQ ID NOs: 2-10 comprises N-terminal and / or C-terminal modifications. In some embodiments, the N-terminal modification is acylation. In some embodiments, the C-terminal modification is amidation.

[0061] In some embodiments, the polypeptides of the present disclosure consist of 20 or fewer amino acids. In some embodiments, a polypeptide comprising the amino acid sequence of SEQ ID NO: 3 further comprises at least one amino acid added to the N-terminus, at least one amino acid added to the C-terminus, or at least one amino acid added to both the N-terminus and the C-terminus. In these embodiments, additions made within 5 amino acids of each terminus have an amino acid sequence that is less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, 0% (or any value or sub-range therebetween) identical to the contiguous amino acid sequence of SEQ ID NO: 1. In these embodiments, additions made within 5 amino acids of each terminus have an amino acid sequence that is less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, 0% (or any value or sub-range therebetween) identical to the contiguous amino acid sequence of SEQ ID NO: 2. In these embodiments, additions made within 5 amino acids of each terminus have an amino acid sequence that is less than 80% identical to the contiguous amino acid sequence of SEQ ID NO: 1 or 2. In some embodiments, additions made within 5 amino acids of each terminus have an amino acid sequence that is less than 60% identical to the contiguous amino acid sequence of SEQ ID NO: 1 or 2. In some, additions made within 5 amino acids of each terminus have an amino acid sequence that is less than 40% identical to the contiguous amino acid sequence of SEQ ID NO: 1 or 2. In some embodiments, additions made within 5 amino acids of each terminus have an amino acid sequence that is less than 20% identical to the contiguous amino acid sequence of SEQ ID NO: 1 or 2. In some embodiments, the polypeptide sequence includes the peptide comprising L-amino acids, the peptide comprising D-amino acids, or the peptide comprising both L-amino acids and D-amino acids.

[0062] In some embodiments, the polypeptide of the present disclosure comprises at least one non-standard amino acid, or the polypeptide comprises two non-standard amino acids. In some embodiments, the non-standard amino acid is ornithine. In some embodiments, the polypeptide further comprises an N-terminal modification, a C-terminal modification, or both an N-terminal and a C-terminal modification. In some embodiments, the N-terminal modification is acylation and / or the C-terminal modification is amidation.

[0063] In some embodiments, the polypeptide comprises a core sequence of ASFTTFTVT.

[0064] In some embodiments, the polypeptide comprises the amino acid sequence of FTTFTVT (SEQ ID NO: 3), KASFTTFTVTKGS (SEQ ID NO: 4), KASFTTFTVTKGS-NH2 (SEQ ID NO: 5), aaEGKASFTTFTVTKGSaa (SEQ ID NO: 6), aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 7), Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8), OASFTTFTVTOS (SEQ ID NO: 9), or OASFTTFTVTOS-NH2 (SEQ ID NO: 10). In some embodiments, the polypeptide consists of an amino acid sequence selected from FTTFTVT (SEQ ID NO: 3), KASFTTFTVTKGS (SEQ ID NO: 4), KASFTTFTVTKGS-NH2 (SEQ ID NO: 5), aaEGKASFTTFTVTKGSaa (SEQ ID NO: 6), aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 7), Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8), OASFTTFTVTOS (SEQ ID NO: 9), or OASFTTFTVTOS-NH2 (SEQ ID NO: 10). In some embodiments, the polypeptide consists of the amino acid sequence of Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8).

[0065] In some embodiments, the modified Cav-1 peptide comprises 1, 2, 3, 4, or more amino acid substitutions, deletions, or insertions relative to the sequence of SEQ ID NO: 1 such that it derives a polypeptide of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 residues.

[0066] The modified Cav-1 peptides provided by the present disclosure exhibit biological activities similar to or the same as the native Cav-1 peptide in in vitro or in vivo assays. In some embodiments, the modified Cav-1 peptide inhibits or blocks apoptosis of lung epithelial cells induced by bleomycin in vitro or in vivo with at least about 20%, or at least about 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, about 95%, 97%, 99%, and any range derivable therein, for example, about 70% - about 80%, about 81% - about 90%, or about 91% - about 99% of the activity of the native Cav-1 peptide. The modified Cav-1 peptide may have an activity of 100% or even higher than that of the native Cav-1 peptide. Assays for testing biological activities, such as anti-fibrotic activity, the ability to affect the expression of mRNA of uPA, uPAR, and PAI-1, or the ability to inhibit the proliferation of lung fibroblasts, are well known in the art.

[0067] The modified Cav-1 peptides of the present disclosure are fragments, derivatives, or variants of the native Cav-1 peptides. The peptides can be synthetic, recombinant, or chemically modified peptides isolated or generated using methods well known in the art. The modifications can be made to the amino acids at the N-terminus, C-terminus, or internally. The peptides can include conservative or non-conservative amino acid changes, as described below. Changes in the polynucleotide can result in amino acid substitutions, additions, deletions, fusions, and truncations in the Cav-1 peptide encoded by the reference sequence. The peptides can also include insertions and substitutions of amino acids (and other molecules) that are not normally present in the peptide sequence that serves as the basis for the modified variant, such as, for example, but not limited to, L-amino acids not normally present in human proteins, or insertions of non-standard amino acids such as ornithine, as well as insertions, deletions, or substitutions of amino acids.

[0068] A. Substitution In some embodiments, the modified Cav-1 peptide comprises one or more conservative amino acid substitutions. Conservative amino acid substitutions result from replacing one amino acid with another having similar structural and / or chemical properties, such as replacing leucine with isoleucine or valine, aspartic acid with glutamic acid, or threonine with serine. Thus, a conservative substitution of a particular amino acid sequence refers to a substitution of an amino acid that is not important for peptide activity, or a substitution of an amino acid with another amino acid having similar properties (e.g., acidic, basic, positively or negatively charged, polar or nonpolar), such that even a substitution of an important amino acid does not reduce the activity of the peptide. Tables of conservative substitutions that provide functionally similar amino acids are well known in the art. For example, the following six groups each contain 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 change, add, or delete a single amino acid or a small percentage of amino acids can also be considered conservative substitutions if the activity of the peptide is not reduced by the change. Insertions or deletions are typically in the range of about 1 to 6 amino acids.

[0069] In some embodiments, an amino acid to substitute an existing amino acid can be selected based on the position of the existing amino acid, i.e., its exposure to the solvent (i.e., whether the amino acid is exposed to the solvent or is present on the outer surface of the peptide or peptide as compared to an amino acid that is not exposed to the solvent and is located internally). The selection of such conservative amino acid substitutions is well known to those skilled in the art, as disclosed, for example, in Dordo et al, J. Mol Biol, 1999, 217, 721-739 and Taylor et al, J. Theor. Biol. 119 (1986); 205-218, as well as S. French and B. Robson, J. Mol. Evol. 19 (1983) 171. Thus, suitable conservative amino acid substitutions can be selected for amino acids outside the protein or peptide (i.e., amino acids exposed to the solvent), for example, but not limited to, the following substitutions can be used: Y with F, T with S or K, P with A, E with D or Q, N with D or G, R with K, G with N or A, T with S or K, D with N or E, I with L or V, F with Y, S with T or A, R with K, G with N or A, K with R, A with S, K or P.

[0070] In some embodiments, suitable conservative amino acid substitutions can be selected for amino acids inside the protein or peptide. For example, suitable conservative substitutions can be used for amino acids inside the protein or peptide (i.e., the amino acids are not exposed to the solvent), for example, but not limited to, the following conservative substitutions can be used: Y is substituted with F, T is substituted with A or S, I is substituted with L or V, W is substituted with Y, M is substituted with L, N is substituted with D, G is substituted with A, T is substituted with A or S, D is substituted with N, I is substituted with L or V, F is substituted with Y or L, S is substituted with A or T, and A is substituted with S, G, T or V. In some embodiments, non-conservative amino acid substitutions are also included within the term variant.

[0071] In some embodiments, the amino acid substitutions can be made at one or more positions in the peptide, where the substitutions are with amino acids having a similar degree of hydrophilicity. The importance of the hydrophobicity index of amino acids in conferring an interactive biological function to a protein is generally understood in the art. It has been recognized that the relative hydrophobicity characteristics of amino acids contribute to the secondary structure of the resulting protein, which in turn defines the interaction of the protein with other molecules, such as enzymes, substrates, receptors, DNA, antibodies, antigens, and the like. Thus, such conservative substitutions can be made in the peptide and are likely to have only a minor effect on their activity. As detailed in U.S. Patent No. 4,554,101, the following hydrophilicity values are 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). These values can be used as a guide, and thus substitutions of amino acids with hydrophilicity values within ±2 are preferred, substitutions of amino acids within ±1 are particularly preferred, and substitutions of amino acids within ±0.5 are even more particularly preferred. Thus, any of the Cav-1 peptides described herein may be modified by substituting an amino acid with a different but similar hydrophilicity value amino acid. Amino acids having a hydrophilicity within + / - 1.0 point, or within + / - 0.5 point, are considered homologous.

[0072] In some embodiments, the modified Cav-1 peptide comprises non-natural amino acids. In some embodiments, the modified Cav-1 peptide comprises a combination of natural and non-natural amino acids or comprises only non-natural amino acids. Non-natural amino acids can include synthetic non-natural amino acids, substituted amino acids, or one or more D-amino acids (or other components of the formulation, excluding protease recognition sequences) as may be desirable in certain circumstances. D-amino acid-containing peptides exhibit increased stability in vitro or in vivo as compared to their L-amino acid-containing forms. Thus, the construction of peptides incorporating D-amino acids can be particularly useful when higher in vivo or intracellular stability is desired or required. More specifically, D-peptides are resistant to endogenous peptidases and proteases, thereby providing better oral trans-epithelial and transdermal delivery of conjugated drugs and conjugates, improved bioavailability of membrane permanent complexes, and long-term intravascular and interstitial lifetimes (when such properties are desired). Further, D-peptides are less likely to induce a humoral immune response throughout the organism because they cannot be efficiently processed for major histocompatibility complex class II-restricted presentation to T helper cells.

[0073] In addition to the 20 “standard” L - amino acids, D - amino acids, non - standard modified amino acids, or amino acids different from the normal ones, which are clearly defined in the art, 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 - amino - isobutyric 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 (Phg); 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 (Thic); homoarginine (hArg); N - acetyllysine (AcLys); 2,4 - diaminobutyric acid (Dbu; Dab); p - aminophenylalanine (Phe(pNH2)); N - methylvaline (MeVal); homocysteine (hCys), homophenylalanine (hPhe), and homoserine (hSer); hydroxyproline (Hyp), homoproline (hPro), N - methylated amino acids, and peptoids (N - substituted glycines).

[0074] A peptide or peptide region has a certain percentage (e.g., 80%, 85%, 90%, or 95%) of "sequence identity" or "homology" to another sequence, which means that when aligned, that percentage of bases (or amino acids) of the two sequences is the same. This alignment and the percent homology or sequence identity can be determined using software programs known in the art, such as the programs described in Current Protocols In Molecular Biology (F.M. Ausubel et al., eds., 1987) Supplement 30, Section 7.7.18, Table 7.7.1. Default parameters can be used for the alignment.

[0075] B. Derivatives In some embodiments, the modified Cav-1 peptide is a derivative of the native Cav-1 peptide. As used herein, the term "derivative" refers to a Cav-1 peptide that has been chemically modified 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 cyclic form, e.g., as a cyclic peptide or 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 if it contains additional chemical moieties that are not normally part of the molecule. Such moieties can alter the pH, or improve the solubility, absorption, biological half-life, etc. of the molecule. Alternatively, this moiety can have the potential to reduce the toxicity of the molecule, eliminate or reduce any undesirable side effects of the molecule, etc. Moieties that can mediate such effects are disclosed in Remington’s Pharmaceutical Sciences, 18th edition, A.R. Gennaro, Ed., Mack Publ., Easton, PA (1990) (incorporated herein by reference in its entirety).

[0076] The term "functional", when used in combination with "derivative" or "variant", refers to a peptide of the invention having a biological activity (either functional or structural) that is substantially similar to the biological activity of an entity or molecule that is a functional derivative or functional variant of the peptide of the invention. The term "functional derivative" is intended to include fragments, analogs, or chemical derivatives of the molecule.

[0077] In some embodiments, the modified Cav-1 peptide may include co-translational and post-translational modifications (e.g., C-terminal peptide cleavage), such as disulfide bond formation, glycosylation, acetylation, phosphorylation, proteolytic cleavage (e.g., cleavage by furin or a metalloprotease), provided that such modifications do not affect the function of the modified Cav-1 peptide.

[0078] In some embodiments, the modified Cav-1 peptide can be a "retro-inverso peptide". A "retro-inverso peptide" refers to a peptide in which the direction of the peptide bond is reversed at at least one position, that is, a peptide in which the amino terminus and the carboxy terminus are reversed with respect to the side chains of the amino acids. Thus, a retro-inverso analog has an inversion of the termini and a reversal of direction of the peptide bond while approximately maintaining the topology of the side chains as in the native peptide sequence. A retro-inverso peptide can contain L-amino acids or D-amino acids, or a mixture of L-amino acids and D-amino acids, up to all of the amino acids being D-isomers. A partially retro-inverso peptide analog is a peptide in which only a portion of the sequence is reversed and replaced with enantiomeric amino acid residues. Since the retro-inverted portion of such an analog has an inverted amino terminus and carboxyl terminus, the amino acid residues adjacent to the retro-inverted portion are each replaced with a-substituted geminal-diaminomethane and malonic acid with similar side chains. The retro-inverso form of a cell-penetrating peptide has been found to function as efficiently as the native form when moving 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, G.C, J.Chem.Soc.Perkin Trans. 1:697-701(1985), and U.S. Patent No. 6,261,569, which are hereby incorporated by reference in their entirety. A process for the solid-phase synthesis of partially retro-inverso peptide analogs is described (EP97994-B), which is also hereby incorporated by reference in its entirety.

[0079] C. Terminal Modification In some embodiments, the Cav-1 peptides of the present disclosure are modified at their amino or carboxy termini (in the case of linear forms). Examples of amino-terminal modifications include, for example, N-glycosylation, N-alkylation, N-acetylation, or N-acylation amino acids. Terminal modifications can include pegylation. An example of a carboxy-terminal modification is a C-terminal amidated amino acid. In some embodiments, the peptide is cross-linked or has a cross-linking site (e.g., the modified Cav-1 peptide has cysteinyl residues and thus forms cross-linked dimers in vitro or in vivo). In some embodiments, one or more peptide bonds are replaced by non-peptide bonds, the N-terminus or C-terminus is replaced, and individual amino acid moieties are modified by treatment with an agent capable of reacting with selected side chains or terminal residues, etc. Either or both of the C-terminus or N-terminus of the amino acid sequence can be linked to a carboxylic acid functional group or an amine functional group, respectively. In some embodiments, the modified Cav-1 peptide includes an N-terminal modification. In some embodiments, the modified Cav-1 peptide includes a C-terminal modification. In some embodiments, the modified Cav-1 peptide includes both N-terminal and C-terminal modifications.

[0080] Non-limiting and exemplary 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 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-, but are not limited thereto. Examples of alkoxycarbonyl and aryloxycarbonyl groups include CH3-O-CO-, (ethyl)-O-CO-, n-propyl-O-CO-, iso-propyl-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-, but are not limited thereto. To facilitate N-acylation, 1 to 4 glycine residues may be present at the N-terminus of the molecule.

[0081] Carboxy-terminal modifications include acylation with carboxylic acids, namely 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 (i) acylation with carboxylic acids, namely formic acid, acetic acid, propionic acid, fatty acids (such as myristic acid, palmitic acid, stearic acid), succinic acid, benzoic acid; (ii) carbonylation (such as benzyloxycarbonylation (Cbz)); (iii) biotinylation; (iv) amidation; (v) binding of dyes such as fluorescein (FITC, FAM, etc.), 7-hydroxy-4-methylcoumarin-3-acetic acid, 7-hydroxycoumarin-3-acetic acid, 7-methoxycoumarin-3-acetic acid, and other coumarins; rhodamine (5-carboxyrhodamine 110 or 6G, 5(6)-TAMRA, ROX); N-[4-(4-dimethylamino)phenylazo]benzoic acid (Dabcyl), 2,4-dinitrobenzene (Dnp), 5-dimethylaminonaphthalene-1-sulfonic acid (Dansyl), and other dyes; and (vi) pegylation, but are not limited thereto.

[0082] The carboxyl group at the C-terminus of the peptide can be protected by groups including, but not limited to, amides (i.e., the hydroxyl group at the C-terminus is replaced by -NH2, -NHR2, and -NR2R3) or esters (i.e., the hydroxyl group at the C-terminus is replaced by -OR2). R2 and R3 are optionally and independently aliphatic, substituted aliphatic, benzyl, substituted benzyl, aryl, or substituted aryl groups. Additionally, R2 and R3 can optionally, together with the nitrogen atom, form a C4-C8 heterocycle having about 0 to 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, -NH2, -NHCH3, -N(CH3)2, -NH(ethyl), -N(ethyl)2, -N(methyl)(ethyl), -NH(benzyl), -N(C1-C4 alkyl)(benzyl), -NH(phenyl), -N(C1-C4 alkyl)(phenyl), -OCH3, -O-(ethyl), -O-(n-propyl), -O-(n-butyl), -O-(iso-propyl), -O-(sec-butyl), -O-(t-butyl), -O-benzyl, and -O-phenyl.

[0083] D. Side Chain Modification In some embodiments, the modified Cav-1 peptides of the present disclosure include modified amino acid side chains. Non-limiting examples of modifications include carboxymethylation, acylation, phosphorylation, glycosylation, or fatty acid acylation. Optionally, an ether bond can be used to link a serine or threonine hydroxyl to a hydroxyl of a sugar. Optionally, an amide bond can be used to link a glutamic acid or aspartic acid carboxyl group 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)). Optionally, acetal and ketal bonds can also be formed between an amino acid and a carbohydrate. Optionally, a fatty acid acyl derivative can be made, for example, by acylation of a free amino group (e.g., lysine) (Toth et al., Peptides: Chemistry, Structure and Biology, Rivier and Marshal, eds., ESCOM Publ., Leiden, 1078-1079 (1990)).

[0084] As used herein, the term "chemical modification" when referring to a modified Cav-1 peptide of the present disclosure refers to a peptide in which at least one of its amino acid residues has been modified by either a natural process such as processing or other post-translational modification, or by chemical modification techniques well known in the art. Numerous known examples of modifications typically include, but are not limited to, acetylation, acylation, amidation, ADP-ribosylation, glycosylation, GPI anchor formation, covalent attachment of a lipid or lipid derivative, methylation, myristoylation, pegylation, prenylation, phosphorylation, ubiquitination, or any similar process.

[0085] Other types of modifications optionally include the addition of cycloalkane moieties to biomolecules such as proteins as described in PCT Application No. WO2006 / 050262, which is hereby incorporated by reference in its entirety. These moieties are designed for use in biomolecules and may optionally be used to confer various properties to the protein.

[0086] Furthermore, optionally, any point on the protein may be modified. For example, as described in PCT Application No. WO2006 / 050247, which is hereby incorporated by reference in its entirety, pegylation of glycosylated moieties on the protein may optionally be carried out. Optionally, one or more polyethylene glycol (PEG) groups may be added to O-linked and / or N-linked glycosylation. The PEG groups may optionally be branched or linear. Optionally, any type of water-soluble polymer may be attached to the glycosylation site on the protein via a glycosyl linker.

[0087] Covalent modifications of the modified Cav-1 peptides of the present disclosure are included within the scope of the invention. Other types of covalent modifications are introduced into the molecule by reacting a target amino acid residue with an organic derivatizing agent capable of reacting with a selected side chain or N-terminal or C-terminal residue.

[0088] Most commonly, cysteinyl residues are reacted with α-haloacetates (and corresponding amines) such as chloroacetic acid or chloroacetamide to obtain carboxymethyl or carboxamidomethyl derivatives. Cysteinyl residues are also derivatized by reaction with bromotrifluoroacetone, α-bromo-β-(5-imidazolyl)propionic acid, chloroacetyl phosphate, N-alkylmaleimide, 3-nitro-2-pyridyldisulfide, methyl 2-pyridyldisulfide, p-chloromercurybenzoic acid, 2-chloromercury-4-nitrophenol, or chloro-7-nitrobenzo-2-oxa-1,3-diazole.

[0089] Histidyl residues are derivatized by reaction with diethyl pyrocarbonate at pH 5.5 - 7.0, since this agent is relatively specific for the histidyl side chain. Para-bromophenacyl bromide is also useful, and this reaction is carried out at pH 6.0 in 0.1 M sodium cacodylate in some embodiments.

[0090] Lysyl and amino-terminal residues are reacted with succinic acid or other carboxylic acid anhydrides. Derivatization with these agents has the effect of inverting the charge of the lysyl residue. Other suitable reagents for derivatizing α-amino-containing residues include imido esters such as methyl picolinimidate, pyridoxal phosphate, pyridoxal, chloroborohydride, trinitrobenzenesulfonic acid, O-methylisourea, 2,4-pentanedione, and transaminase-catalyzed reactions with glyoxylic acid.

[0091] Arginyl residues are modified by reaction with one or several conventional reagents such as phenylglyoxal, 2,3-butanedione, 1,2-cyclohexanedione, and ninhydrin.

[0092] Derivatization of arginine residues requires that the reaction be carried out under alkaline conditions due to the high pKa of the guanidine functional group. Furthermore, these reagents can react with lysine groups and the arginine epsilon-amino group.

[0093] Specific modification of tyrosyl residues may be carried out, particularly for the purpose of introducing spectral labels to 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 iodinated using 125I or 131I to prepare labeled peptides for use in radioimmunoassays.

[0094] Carboxyl side chains (aspartyl or glutamyl) are selectively modified by reaction with carbodiimides (R-N=C=N-R'), such as 1-cyclohexyl-3-(2-morpholinyl-4-ethyl) carbodiimide or 1-ethyl-3-(4-azonia-4,4-dimethylpentyl) carbodiimide, where R and R' are various alkyl groups. Further, aspartyl and glutamyl residues are converted to asparaginyl and glutaminyl residues by reaction with ammonium ions.

[0095] Derivatization with bifunctional agents is useful for cross-linking to a water-insoluble support matrix or surface for use in a method of purifying an anti-CHF antibody, and vice versa. Commonly used cross-linking agents include, for example, 1,1-bis(diazoacetyl)-2-phenylethane, glutaraldehyde, N-hydroxysuccinimide esters such as esters with 4-azidosalicylic acid, homobifunctional imide esters including disuccinimidyl esters such as 3,3'-dithiobis(succinimidyl propionate), and bifunctional maleimides such as bis-N-maleimide-1,8-octane. Derivatizing agents such as methyl-3-[(p-azidophenyl)dithio]propionimidate produce photoexcitable intermediates that can form cross-links in the presence of light. Alternatively, water-insoluble reactive matrices and reactive substrates such as cyanogen bromide-activated carbohydrates described in U.S. Pat. Nos. 3,969,287, 3,691,016, 4,195,128, 4,247,642, 4,229,537, and 4,330,440 are used for protein immobilization.

[0096] Glutaminyl and asparaginyl residues are often 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 present invention.

[0097] Other modifications include hydroxylation of proline and lysine, phosphorylation of the hydroxyl group of a seryl or threonyl residue, methylation of the α-amino group of the lysine, arginine, and histidine side chains (T.E. Creighton, 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.

[0098] E. Capping In some embodiments, the modified Cav-1 peptide has its N-terminus and C-terminus capped with acyl (abbreviated "Ac") and amide (abbreviated "Am") groups, respectively (e.g., N-terminal acetyl (CH3CO-) and C-terminal amide (-NH2)). In some embodiments, the modified Cav-1 peptide has its N-terminus capped with an acyl group (e.g., N-terminal acetyl (CH3CO-)). In some embodiments, the modified Cav-1 peptide has its C-terminus capped with an amide group (e.g., C-terminal amide (-NH2)).

[0099] In some embodiments, the modified Cav-1 peptide has its N-terminus capped. A wide range of N-terminal capping functions, such as attachment to the terminal amino group, e.g., formyl, alkanoyl having 1 to 10 carbon atoms such as acetyl, propionyl, butyryl, 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-hexa-3-enoyl, Substituted alkynoyl having 1 to 10 carbon atoms such as 3-hydroxy-hexa-5-ynoyl, Substituted aroyl such as 4-chlorobenzoyl or 8-hydroxy-naphth-2-oyl, 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) (wherein 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) (wherein R' is alkanoyl, alkenoyl, alkynoyl, aroyl, heteroaroyl, substituted alkanoyl, substituted alkenoyl, substituted alkynoyl, substituted aroyl, or substituted heteroaroyl (as defined above in all cases)) are contemplated.

[0100] In some embodiments, the modified Cav-1 peptide has its C-terminus capped. The C-terminus capping function can be either an amide bond or an ester bond with the terminal carboxyl. The capping function providing an amide bond is represented as NR 1 R 2 wherein R 1 and R2 may be independently selected from the following group: hydrogen, alkyl having from 1 to 10 carbon atoms such as methyl, ethyl, isopropyl, etc., alkenyl having from 1 to 10 carbon atoms such as prop-2-enyl, etc., alkynyl having from 1 to 10 carbon atoms such as prop-2-ynyl, etc., substituted alkyl having from 1 to 10 carbon atoms such as hydroxyalkyl, alkoxyalkyl, mercaptoalkyl, alkylthioalkyl, halogenoalkyl, cyanoalkyl, aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl, alkanoylalkyl, carboxyalkyl, carbamoylalkyl, etc., substituted alkenyl having from 1 to 10 carbon atoms such as hydroxyalkenyl, alkoxyalkenyl, mercaptoalkenyl, alkylthioalkenyl, halogenoalkenyl, cyanoalkenyl, aminoalkenyl, alkylaminoalkenyl, dialkylaminoalkenyl, alkanoylalkenyl, carboxyalkenyl, carbamoylalkenyl, etc., substituted alkynyl having from 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, etc., aryl such as phenyl or 1-naphthyl, etc., heteroaryl such as 4-quinolyl, etc., alkanoyl having from 1 to 10 carbon atoms such as acetyl or butyryl, etc., aroyl such as benzoyl, etc., heteroaroyl such as 3-quinolinoyl, etc., OR’ or NR’R’’ (wherein R’ and R’’ are independently hydrogen, alkyl, aryl, heteroaryl, acyl, aroyl, sulfonyl, sulfinyl), or SO2-R’’’ or SO-R’’’ (wherein R’’’ is substituted or unsubstituted alkyl, aryl, heteroaryl, alkenyl, or alkynyl).

[0101] The capping function that provides an ester bond is represented as OR, wherein R can be alkoxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, substituted alkoxy, substituted aryloxy, substituted heteroaryloxy, substituted aralkyloxy, or substituted heteroaralkyloxy.

[0102] In some embodiments, the N-terminal or C-terminal capping function, or both, is such that the capped molecule functions as a prodrug (a pharmacologically inactive derivative of the parent drug molecule) that undergoes spontaneous or enzymatic conversion in the body to release the active drug, and has a structure such that its delivery characteristics are improved compared to the parent drug molecule (Bundgaard H, Ed: Design of Prodrugs, Elsevier, Amsterdam, 1985).

[0103] A judicious choice of capping group enables the addition of other activities to the peptide. For example, the presence of a sulfhydryl group linked to the N-terminal or C-terminal cap allows the derivatized peptide to be conjugated to other molecules.

[0104] F. Multimerization Embodiments of the present disclosure also include longer polypeptides constructed from repeating units of modified Cav-1 peptides. In some embodiments, the peptide multimer includes different combinations of peptides. In some embodiments, the multimeric peptide is made by chemical synthesis or by recombinant DNA techniques as discussed herein. When produced by chemical synthesis, in some embodiments, the oligomer has 2 to 5 repeats of the core peptide sequence, and the total number of amino acids in the multimer should not exceed about 160 residues or should be 100 residues or less (or their equivalents, including linkers or spacers).

[0105] In some embodiments, the modified Cav-1 peptide is a multimer comprising at least two peptides of the present disclosure. In some embodiments, the first peptide of the at least two peptides is substantially identical to the second peptide of the at least two peptides. In some embodiments, the first peptide of the at least two peptides is not identical to the second peptide of the at least two peptides.

[0106] G. Peptide mimetics In some embodiments, the modified Cav-1 peptide is a peptide mimetic compound that mimics the biological effects of the native Cav-1 peptide. In some embodiments, the peptide mimetic is a non-natural peptide or non-peptide agent that reproduces the steric properties of the binding elements of the native Cav-1 peptide so as to have the binding activity and biological activity of the native Cav-1 peptide. Similar to the native Cav-1 peptide or peptide multimer, the peptide mimetic has a binding surface (which interacts with any ligand to which the native Cav-1 peptide binds) and a non-binding surface.

[0107] In some embodiments, the disclosure also includes modified Cav-1 peptides that retain some of the peptide characteristics. For example, any proteolytically labile bonds within the Cav-1 peptides of the invention can be selectively replaced by non-peptide elements such as isosteres (N-methylation, D-amino acids) or reduced peptide bonds while the rest of the molecule retains its peptidic nature.

[0108] For several bioactive peptides / peptides such as opioid peptides, VIP, thrombin, HIV protease, peptidomimetic compounds that are either agonists, substrates or inhibitors have been described. 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 structure are described in Johnson et al., In: Biotechnology and Pharmacy, Pezzuto et al., Chapman and Hall (Eds.), NY, 1993. Using these methods, peptidomimetics are made that possess at least the binding ability and specificity of the native Cav-1 peptide and, in some embodiments, also possess biological activity. The knowledge of peptide chemistry and general organic chemistry available to those of skill in the art is sufficient in light of the present disclosure for the design and synthesis of such compounds.

[0109] For example, such peptidomimetics can be identified by examination of the three-dimensional structure of the peptides of the invention, whether free or complexed and bound to a ligand (e.g., soluble uPAR or a fragment thereof). Alternatively, the structure of the peptides of the invention bound to their ligand can be obtained by techniques of nuclear magnetic resonance spectroscopy. Deeper knowledge of the stereochemistry of the interaction of the peptide with its ligand or receptor will allow for the rational design of such peptidomimetic agents. The structure of the peptides of the invention or the peptides in the absence of ligand can also provide a scaffold for the design of mimetic molecules.

[0110] H.PEGylation In some embodiments, the modified Cav-1 peptides of the disclosure are conjugated to heterologous peptide segments or polymers such as polyethylene glycol. In some embodiments, the modified Cav-1 peptides are linked to PEG to increase the hydrodynamic radius of the enzyme, thereby increasing persistence in serum. In some embodiments, the modified Cav-1 peptides are conjugated to any targeting agent such as a ligand having the ability to specifically and stably bind to an external receptor (see, e.g., U.S. Patent Publication No. 2009 / 0304666).

[0111] In some embodiments, the disclosure provides methods and formulations related to the PEGylation of Cav-1 peptides. PEGylation is the process of covalently attaching poly(ethylene glycol) polymer chains to another molecule, typically a drug or therapeutic protein. PEGylation is conventionally accomplished by incubating a reactive derivative of PEG with the target macromolecule. Covalent attachment of PEG to a drug or therapeutic protein can "mask" the agent from the host immune system (reducing immunogenicity and antigenicity) or increase the hydrodynamic size (size in solution) of the agent, thereby extending circulation time by decreasing renal clearance. PEGylation can also render hydrophobic drugs and proteins water-soluble.

[0112] The first step in PEGylation is the suitable functionalization of the PEG polymer at one or both ends. PEG that is activated at each end with the same reactive moiety is known as "homo-bifunctional," while if the functional groups present are different, the PEG derivative is termed "hetero-bifunctional" or "heterofunctional." To attach the PEG to the desired molecule, a chemically active or activated derivative of the PEG polymer is prepared.

[0113] The selection of a suitable functional group for the PEG derivative is based on the type of available reactive groups on the modified Cav-1 peptide to which the PEG is to be attached. In the case of proteins, typical reactive amino acids include lysine, cysteine, histidine, arginine, aspartic acid, glutamic acid, serine, threonine, and tyrosine. The N-terminal amino group and the C-terminal carboxylic acid group can also be used.

[0114] The techniques used to form first-generation PEG derivatives generally involve reacting the PEG polymer with groups that react with hydroxyl groups, typically anhydrides, acid chlorides, chloroformates, and carbonates. In second-generation PEGylation chemistry, more efficient functional groups such as aldehydes, esters, and amides are made available for conjugation.

[0115] As the applications of PEGylation become increasingly sophisticated and advanced, the need for hetero-bifunctional PEGs for conjugation is growing. These hetero-bifunctional PEGs are very useful for linking two entities where a hydrophilic, flexible, and biocompatible spacer is required. Preferred end groups for hetero-bifunctional PEGs are maleimide, vinyl sulfone, pyridyldisulfide, amine, carboxylic acid, and N-hydroxysuccinimide (NHS) ester.

[0116] The most common modifiers, or linkers, are based on methoxy PEG (mPEG) molecules. Their activity depends on adding a protein modification group to the alcohol terminus. In some embodiments, polyethylene glycol (PEG diol) is used as the precursor molecule. To create hetero- or homodimeric PEG-linked molecules, the diol is then modified at both ends.

[0117] Proteins are generally PEGylated at nucleophilic sites such as unprotonated thiols (cysteiny residues) or amino groups. Examples of cysteine-specific modification reagents include PEG maleimide, PEG iodoacetic acid, PEG thiol, and PEG vinyl sulfone. All four are strongly cysteine-specific under mild conditions and a pH ranging from neutral to slightly alkaline, but each has some drawbacks. The thioether formed with maleimide can become somewhat unstable under alkaline conditions, so there may be some limitations to the formulation options using this linker. The carbamothioate bond formed with iodo PEG is more stable, but free iodine can modify tyrosine residues under some conditions. PEG thiol forms a disulfide bond with the protein thiol, but this bond can also be unstable under alkaline conditions. The reactivity of PEG-vinyl sulfone is relatively slow compared to maleimide and iodo PEG, but the thioether bond formed is very stable. The slower reaction rate can also make it easier to control the PEG-vinyl sulfone reaction.

[0118] Site-specific PEGylation at native cysteine residues is rarely performed. This is because these residues are usually in the form of disulfide bonds or are required for biological activity. On the other hand, site-directed mutagenesis can be used to incorporate a cysteine PEGylation site for a thiol-specific linker. The cysteine mutation must be designed such that it is available to the PEGylation reagent and remains biologically active after PEGylation.

[0119] Examples of amine-specific modifiers include PEG NHS esters, PEG tresylates, PEG aldehydes, PEG isothiocyanates, and several others. All react under mild conditions and are highly specific for amino groups. PEG NHS esters are perhaps one of the more reactive agents, but their high reactivity can make it difficult to control the PEGylation reaction on a large scale. PEG aldehydes form imines with amino groups, which are then reduced to secondary amines with sodium cyanoborohydride. Unlike sodium borohydride, sodium cyanoborohydride does not reduce disulfide bonds. However, this chemical is highly toxic and must be handled carefully, especially at lower pH where it becomes more volatile.

[0120] Due to multiple lysine residues on most proteins, site-specific PEGylation can be difficult. Since these reagents react with unprotonated amino groups, it is possible to direct PEGylation towards amino groups with lower pK by performing the reaction at a lower pH. Generally, the pK of alpha-amino groups is 1-2 pH units lower than that of epsilon-amino groups of lysine residues. By PEGylating the molecule below pH 7, high selectivity for the N-terminus can be frequently achieved. However, this is only feasible if the N-terminal portion of the protein is not required for biological activity. Still, the pharmacokinetic advantages of PEGylation often outweigh a substantial decrease in in vitro biological activity and result in products with much greater in vivo biological activity, regardless of the PEGylation chemistry.

[0121] There are several parameters to consider when developing a PEGylation procedure. Fortunately, the important parameters are usually four or fewer. A "design of experiments" approach can be very useful for optimizing PEGylation conditions. 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 exclusion of oxygen (oxygen can contribute to intermolecular disulfide formation by the protein, thereby reducing the yield of the PEGylated product). For amine-specific modifications, the same factors should be considered (excluding oxygen), except that pH may be even more important, especially when targeting the N-terminal amino group.

[0122] For both amine-specific and thiol-specific modifications, the reaction conditions can affect the stability of the protein. This can limit temperature, protein concentration, and pH. Additionally, the reactivity of the PEG linker should be known before initiating the PEGylation reaction. For example, if the activity of the PEGylation agent is only 70%, it should be ensured that only the active PEG molecules are counted in the reaction stoichiometry of protein and PEG, based on the amount of PEG used.

[0123] I. Fusion Protein In some embodiments, the present disclosure provides a fusion protein of a modified Cav-1 peptide. For example, the fusion may use a leader sequence from another species to enable recombinant expression of the protein in a heterologous host. The fusion protein may include a half-life extender. Another useful fusion may include the addition of a protein affinity tag, such as a serum albumin affinity tag or six histidine residues, or an immunologically active domain (including cleavable ones), such as an antibody 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 immunologically active domain.

[0124] In some embodiments, the modified Cav-1 peptide is linked to a peptide that extends the in vivo half-life, such as an XTEN® peptide (Schellenberger et al., 2009), an IgG Fc domain, albumin, or an albumin-binding peptide.

[0125] 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 the complete fusion protein or by ligation of DNA sequences encoding the heterologous domains followed by expression of the intact fusion protein.

[0126] Production of a fusion protein that restores the functional activity of the parent protein can be facilitated by connecting the gene with a bridging DNA segment encoding a peptide linker that is spliced between tandemly connected polypeptides. This linker will be of sufficient length to allow proper folding of the resulting fusion protein.

[0127] i) Linker In some embodiments, the modified Cav-1 peptide is chemically conjugated using a bifunctional crosslinking reagent or fused at the protein level with a peptide linker.

[0128] Bifunctional crosslinking reagents are widely used for various purposes, including the preparation of affinity matrices, modification and stabilization of diverse structures, identification of ligand and 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 peptides of the present disclosure.

[0129] Homo-bifunctional reagents bearing two identical functional groups have been found to be very efficient in inducing crosslinks between identical and different macromolecules or subunits of macromolecules and in linking peptide ligands to their specific binding sites. Hetero-bifunctional reagents contain two different functional groups. By taking advantage of the different reactivities of the two different functional groups, crosslinking can be selectively and sequentially controlled. Bifunctional crosslinking reagents can be classified according to the specificity of their functional groups, for example, amino-specific, sulfhydryl-specific, guanidine-specific, indole-specific, carboxyl-specific groups. Among these, reagents directed towards free amino groups are particularly popular because of their commercial availability, ease of synthesis, and mild reaction conditions under which they can be applied.

[0130] Most hetero-bifunctional crosslinking reagents contain a primary amine-reactive group and a thiol-reactive group. In another example, a hetero-bifunctional crosslinking reagent and methods of using this crosslinking reagent are described (U.S. Patent No. 5,889,155, which is incorporated herein by reference in its entirety). The crosslinking reagent combines a nucleophilic hydrazide residue with an electrophilic maleimide residue to enable, in one example, the coupling of an aldehyde to a free thiol. The crosslinking reagent can be modified to crosslink various functional groups.

[0131] Furthermore, for example, the modified Cav-1 peptides of the present disclosure may be conjugated using any other linker / coupling agent and / or mechanism known to those skilled in the art, such as antibody-antigen interactions, avidin-biotin binding, 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.

[0132] In some embodiments, the modified Cav-1 peptides include a crosslinking agent that has reasonable stability in the blood. Numerous types of disulfide bond-containing linkers are known that can be successfully used to conjugate targeting agents and therapeutic / prophylactic agents. Linkers containing sterically hindered disulfide bonds may be found to enhance stability in vivo. Thus, in some embodiments, the modified Cav-1 peptides include a sterically hindered crosslinking agent.

[0133] In addition to hindered crosslinking agents, unhindered linkers can also be used herein. In some embodiments, the modified Cav-1 peptides include an unhindered crosslinking linker. Other useful crosslinking agents that are not thought to contain or generate protected disulfides include SATA, SPDP, and 2-iminothiolane (Wawrzynczak and Thorpe, 1987). The use of such crosslinking agents is well understood in the art.

[0134] In some embodiments, the modified Cav-1 peptides include a flexible linker.

[0135] Once chemically conjugated, the modified Cav-1 peptides are generally purified to separate the conjugate from unconjugated substances and other contaminants. A number of purification techniques are available for use in providing a conjugate of sufficient purity to make it clinically useful.

[0136] Purification methods based on size separation, such as gel filtration, gel permeation, or high performance liquid chromatography, are generally the most useful. Other chromatography techniques, such as blue sepharose separation, may also be used. Conventional methods for purifying fusion proteins from inclusion bodies, such as the use of weak detergents like sodium N-lauroyl-sarcosine (SLS), may be useful.

[0137] ii) Cell-penetrating and membrane-translocating peptides In some embodiments, the modified Cav-1 peptide comprises a cell-binding domain or a cell-penetrating peptide (CPP). As used herein, the terms “cell-penetrating peptide,” “membrane-translocating domain,” and “protein transduction domain” are used interchangeably and refer to a segment of a peptide sequence that enables a 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, HIV-binding peptides, HIV-1 Tat (HIV), Tat-derived peptides, penetratin, VP22-derived or analogous peptides, HSV VP22 (herpes simplex), protegrin I, MAP, KALA or protein transduction domain (PTD), PpT620, proline-rich peptides, arginine-rich peptides, lysine-rich peptides, MPG-peptide(s), Pep-1, L-oligomers, calcitonin peptide(s), Antennapedia-derived peptides (particularly Drosophila Antennapedia-derived), 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 peptides, SAP, or segments derived from histones.

[0138] CPPs typically have an amino acid composition that is either rich in the relative abundance of positively charged amino acids such as lysine or arginine, or contains an alternating pattern of polar / charged and nonpolar hydrophobic amino acids. These two types of structures are referred to as polycationic or amphipathic, respectively. Typically, CPPs are 8 - 50 residue peptides that have the ability to cross cell membranes and enter most cell types. Frankel and Pabo described the ability of the trans - activating transcriptional activator (HIV - TAT) from human immunodeficiency virus 1 to penetrate cells (Frankel, A.D. and C.O. Pabo, Cellular uptake of the tat protein from human immunodeficiency virus. Cell, 1988. 55(6): p. 1189 - 93). In 1991, the transduction of neurons by the Antennapedia homeodomain (DNA - binding domain) from Drosophila melanogaster was also described (Joliot, A., et al., Antennapedia homeobox peptide regulates neural morphogenesis. Proc Natl Acad Sci U S A, 1991. 88(5): p. 1864 - 8). In 1994, the first 16 - mer peptide CPP, called penetratin (RQIKIWFQNRRMKWKK, SEQ ID NO: 113), was characterized from the third helix of the homeodomain of the Drosophila Antennapedia homeobox gene product (Derossi, D., et al., The third helix of the Antennapedia homeodomain translocates through biological membranes. J Biol Chem, 1994. 269(14): p. 10444 - 50). Subsequently, in 1998, the minimal domain of TAT required for protein transduction (e.g., GRKKRRQRRRPPQ, SEQ ID NO: 112) was identified (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 20 years, 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 venoms such as melittin (GIGAVLKVLTTGLPALISWIKRKRQQ, SEQ ID NO: 114) (Dempsey, C. E., The actions of melittin on membranes. Biochim Biophys Acta, 1990. 1031(2): p. 143-61), mastoparan (Konno, K., et al., 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), maurocalcin (Esteve, E., et al., Transduction of the scorpion toxin maurocalcin into cells. Evidence that the toxin crosses the plasma membrane. J Biol Chem, 2005. 280(13): p. 12833-9), crotamine (Nascimento, F. D., et al., Numerous peptides from various origins have been described, including crotamine, which mediates gene delivery into cells through binding to heparan sulfate proteoglycans (J Biol Chem, 2007. 282(29): p. 21349 - 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 poly - arginine (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 - mentioned CPPs can be used in the modified Cav - 1 peptides of the present disclosure. Several other CPPs described by Milletti F. (Drug Discov Today 17(15 - 16): 850 - 60, 2012) can also be used in the modified Cav - 1 peptides of the present disclosure.

[0139] III. Pharmaceutical Formulations In some embodiments, the present disclosure relates to a pharmaceutical formulation comprising a modified Cav - 1 peptide. In some embodiments, the pharmaceutical formulations described herein may be useful for the treatment or prevention of the diseases, injuries, or infections described herein.

[0140] In some embodiments, a pharmaceutical formulation comprising a modified Cav-1 peptide is formulated to be administered intravenously, intrathecally, intradermally, transdermally, intramedullary, intraarterially, intraperitoneally, intranasally, intravaginally, intracellulary, intraarticularly, intralesionally, rectally, intramuscularly, subcutaneously, mucosally, orally, topically, locally, by inhalation (e.g., inhalation of a spray or dry powder formulation), by injection, by infusion, by continuous infusion, by topical perfusion directly immersing target cells, via a catheter, via lavage, in a lipid composition (e.g., liposome), or by other methods known to those of skill in the art or any combination of the foregoing (see, e.g., Remington’s Pharmaceutical Sciences, 18th Ed., 1990, incorporated herein by reference). In some embodiments, the pharmaceutical formulation is formulated to be administered intravenously, intrathecally, subcutaneously and / or intraperitoneally.

[0141] In one aspect, the present disclosure provides a pharmaceutical formulation comprising a modified Cav-1 peptide comprising a core sequence of FTTFTVT and sucrose acetate isobutyrate (SAIB). In some embodiments, the pharmaceutical formulation comprises a modified Cav-1 peptide comprising a core sequence of ASFTTFTVT and SAIB. In some embodiments, SAIB is a mixed ester of sucrose esterified with two acetate groups and six isobutyrate groups, the ester being completely amorphous and having a viscosity of greater than 100,000 cP at 30°C.

[0142] In some embodiments, the formulation comprising a modified Cav-1 peptide is selected from any one of the polypeptides disclosed herein. In some embodiments, the formulation comprises SAIB.

[0143] In some embodiments, SAIB is mixed with a modified Cav-1 peptide to prepare a suspension. In some embodiments, SAIB and the modified Cav-1 peptide are mixed at a temperature from room temperature to about 37°C. In some embodiments, SAIB and the modified Cav-1 peptide are mixed at a temperature of about 37°C or lower. In some embodiments, SAIB and the modified Cav-1 peptide can be mixed with one or more biocompatible solvents. In some embodiments, SAIB and the modified Cav-1 peptide can be mixed with one or more biocompatible solvents selected from water, ethanol, isopropyl alcohol, saline, or phosphate buffered saline (PBS). In some embodiments, the use of different solvents can change the viscosity of the solution of SAIB and the Cav-1 peptide. In some embodiments, the modified Cav-1 peptide formulation is an injectable formulation.

[0144] In some embodiments, the SAIB formulation further comprises a first solvent selected from acetone, benzyl alcohol, butylene glycol, caprolactam, caprolactone, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl lactate, glycerol, glycerol formal, glycol formal, tetraglycol, N-methyl-2-pyrrolidone, polyethylene glycol, methoxypolyethylene glycol, alkoxypolyethylene glycol, propylene carbonate, 2-pyrrolidone, triacetin, triethyl citrate, or combinations thereof. In some embodiments, the first solvent is N-methylpyrrolidone (NMP), absolute ethanol, and / or ethyl acetate. In some embodiments, the first solvent is NMP and / or ethyl acetate.

[0145] In some embodiments, the SAIB is present in the first solvent at about 50% w / w to about 95% w / w (including any value or sub-range therebetween). In some embodiments, the SAIB is present in the first solvent at about 50% w / w, about 51% w / w, about 52% w / w, about 53% w / w, about 54% w / w, about 55% w / w, about 56% w / w, about 57% w / w, about 58% w / w, about 59% w / w, about 60% w / w, about 61% w / w, about 62% w / w, about 63% w / w, about 64% w / w, about 65% w / w, about 66% w / w, about 67% w / w, about 68% w / w, about 69% w / w, about 70% w / w, about 71% w / w, about 72% w / w, about 73% w / w, about 74% w / w, about 75% w / w, about 76% w / w, about 77% w / w, about 78% w / w, about 79% w / w, about 80% w / w, about 81% w / w, 82% w / w, about 83% w / w, about 84% w / w, about 85% w / w, about 86% w / w, about 87% w / w, 88% w / w, about 89% w / w, about 90% w / w, about 91% w / w, about 92% w / w, about 93% w / w, about 94% w / w, or about 95% w / w. In some embodiments, the SAIB is present in the first solvent at about 70% w / w to about 90% w / w (including any value or sub-range therebetween). In some embodiments, the SAIB is present in the first solvent at about 80% w / w.

[0146] In some embodiments, the modified Cav-1 peptides of the present disclosure are micronized. In some embodiments, the modified Cav-1 peptides are micronized prior to admixing with SAIB. In some embodiments, the modified Cav-1 peptides are micronized during the formulation process. In some embodiments, the modified Cav-1 peptide particles are sized by using techniques including, but not limited to, attrition, grinding (e.g., air friction grinding (jet mill), ball mill), coacervation, complex coacervation, high pressure homogenization, spray drying, and / or supercritical fluid crystallization. In some embodiments, the modified Cav-1 peptide particles are sized by mechanical impact (e.g., by a hammer mill, ball mill, and / or pin mill). In some embodiments, the modified Cav-1 peptides are micronized via fluid energy (e.g., by a spiral jet mill, loop jet mill, and / or fluidized bed jet mill). In some embodiments, the modified Cav-1 peptide particles are sized by spray drying. In some embodiments, the modified Cav-1 peptides are micronized by spray drying.

[0147] In some embodiments, the formulations described herein comprise one or more multi-particle modified Cav-1 peptides. In some embodiments, the formulations described herein comprise micronized modified Cav-1 peptides. In some embodiments, the average particle size of the micronized polypeptide ranges from about 0.5 μm to about 500 μm, including any value or sub-range therebetween. In some embodiments, the average particle size of the micronized polypeptide ranges from about 0.5 μm to about 200 μm, including any value or sub-range therebetween. In some embodiments, the average particle size of the micronized polypeptide ranges from about 0.5 μm to about 100 μm, including any value or sub-range therebetween. In some embodiments, the average particle size of the micronized polypeptide ranges from about 1 μm to about 50 μm, including any value or sub-range therebetween. In some embodiments, the average particle size of the micronized polypeptide ranges from about 1 μm to about 5 μm, including any value or sub-range therebetween. In some embodiments, the micronized polypeptide has an average particle size of less than about 10 μm, less than about 5 μm, or less than about 3 μm. In some embodiments, it is micronized. The peptide has an average particle size of about 1.0 μm, about 1.1 μm, about 1.2 μm, about 1.3 μm, about 1.4 μm, about 1.5 μm, about 1.6 μm, about 1.7 μm, about 1.8 μm, about 1.9 μm, about 2.0 μm, about 2.1 μm, about 2.2 μm, about 2.3 μm, about 2.4 μm, about 2.5 μm, about 2.6 μm, about 2.7, about 2.8 μm, about 2.9 μm, about 3.0 μm, about 3.1 μm, about 3.2 μm, about 3.3 μm, about 3.4 μm, about 3.5 μm, about 3.6 μm, about 3.7 μm, about 3.8 μm, about 3.9 μm, about 4.0 μm, about 4.1 μm, about 4.2 μm, about 4.3 μm, about 4.4 μm, about 4.5 μm, about 4.6 μm, about 4.7 μm, about 4.8 μm, about 4.9 μm, or about 5.0 μm.

[0148] In some embodiments, SAIB and the modified Cav-1 peptide can be combined by any suitable means known in the art, such as direct mixing, extrusion coating, spray drying, blending, and encapsulation. In some embodiments, SAIB and the modified Cav-1 peptide are combined as described in Example 1 and formulated into a pharmaceutical preparation.

[0149] In some embodiments, the preparation is in the form of an emulsion, solution, microsphere, nanoparticle, nanosphere, implant, or gel. In some embodiments, the preparation is a suspension. In some embodiments, the preparation further comprises a second solvent. In some embodiments, the second solvent is sterile water, phosphate buffered saline (PBS), or any pharmaceutically acceptable diluent known in the art. In some embodiments, the second solvent is PBS. In some embodiments, the SAIB preparation is in the form of an emulsion, solution, microsphere, nanoparticle, nanosphere, implant, or gel.

[0150] In one aspect, the present disclosure provides a pharmaceutical preparation comprising a modified Cav-1 peptide comprising the core sequence of FTTFTVT and a biodegradable polymer.

[0151] In some embodiments, the formulations comprising the modified Cav-1 peptides described herein are parenteral depot formulations or in situ forming parenteral systems (ISFIs). In some embodiments, the formulation comprises a mixture of a modified Cav-1 peptide dissolved or suspended in a pharmaceutically acceptable water-miscible organic solvent and a biodegradable polymer(s). The formulation forms and methods of ISFIs are further described in Musmade et al., J. Biol. Chem. Chron., 2019, 5(1), 14-21, Ko et al., Progress in Polymer Science, Vol. 38, 2013, and Kanwar and Sinha, Crit. Rev. Ther. Drug Carrier Systems, Vol. 36, 2019, which are hereby incorporated by reference in their entirety.

[0152] In some embodiments, the formulations comprising the modified Cav-1 peptides described herein comprise a biodegradable polymer. In some embodiments, the biodegradable polymer is poly(lactic acid), poly(glycolic acid), poly(lactic-co-glycolic acid), poly(ethylene glycol), poly(vinyl alcohol), polysiloxane, poly(ethylene-vinyl acetate), polyurethane, polyalkyl cyanoacrylate, poly-ε-caprolactone, or a hydrogel polymer.

[0153] In some embodiments, the formulation comprises a modified Cav-1 peptide and SAIB in a weight ratio of about 1:50 to about 1:1, about 1:50 to about 1:5, about 1:50 to about 1:10, or any value or sub-range therebetween. In some embodiments, the weight ratio of the modified Cav-1 peptide to SAIB is about 1:40 to about 1:1 or about 1:40 to about 1:5 (including any value or sub-range therebetween). In some embodiments, the weight ratio of the modified Cav-1 peptide to SAIB is about 1:30 to about 1:1 or 1:30 to about 1:5 (including any value or sub-range therebetween). In some embodiments, the weight ratio of the modified Cav-1 peptide to SAIB is about 1:20 to about 1:1 or about 1:20 to about 1:5 (including any value or sub-range therebetween). In some embodiments, the weight ratio of the modified Cav-1 peptide to SAIB is about 1:30, about 1:29, about 1:28, about 1:27, about 1:26, about 1:25, about 1:24, about 1:23, about 1:22, about 1:21, about 1:20, about 1:19, about 1:18, about 1:17, about 1:16, about 1:15, about 1:14, about 1:13, about 1:12, about 1:11, about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, or any value or sub-range therebetween. In some embodiments, the weight ratio of the modified Cav-1 peptide to SAIB is about 1:50. In some embodiments, the weight ratio of the modified Cav-1 peptide to SAIB is about 1:40. In some embodiments, the weight ratio of the modified Cav-1 peptide to SAIB is about 1:30. In some embodiments, the weight ratio of the modified Cav-1 peptide to SAIB is about 1:17. In some embodiments, the weight ratio of the modified Cav-1 peptide to SAIB is about 1:13. In some embodiments, the weight ratio of the modified Cav-1 peptide to SAIB is about 1:10. In some embodiments, the weight ratio of the modified Cav-1 peptide to SAIB is about 1:8. In some embodiments, the weight ratio of the modified Cav-1 peptide to SAIB is about 1:5. In some embodiments, the weight ratio of the modified Cav-1 peptide to SAIB is about 1:1.

[0154] In some embodiments, the formulation further comprises any molecule or material that functions to minimize chemical degradation of the modified Cav-1 peptide and / or maintains the physicochemical stability of the formulation. Exemplary molecules or materials include amphiphilic molecules such as mono-C 12 - 18 sodium alkyl sulfate salts, dialkyl ester sulfosuccinate derivatives having from 3 to 16 carbon atoms, dioctyl sulfosuccinate, benzenesulfonic acid, naphthalene-1,5-disulfonic acid, camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, dodecyl sulfate, para-toluenesulfonic acid, naphthalene-2-sulfonic acid, cholesterol sulfate, heptanesulfonic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, oleic acid, palmitic acid, pamoic acid, benzoic acid, stearic acid, undecylenic acid, and phospholipids; other polymers such as olylactide, polyglycolide, polycaprolactone, polyanhydrides, polyamines, polyurethanes, polyesteramides, polyorthoesters, polydioxanone, polyacetals, polyketals, polycarbonates, polyphosphoesters, polyoxaesters, polyorthocarbonates, polyphosphazenes, succinates, poly(malic acid), poly(amino acids), polyvinylpyrrolidone, polyethylene glycol, polyhydroxycellulose, chitin, chitosan, hyaluronic acid and copolymers, terpolymers and mixtures thereof; and free radical scavengers and stabilizers such as cysteine or methionine, d-alpha tocopherol acetate, dl-alpha tocopherol, ascorbyl palmitate, butylated hydroxyanisole, butyl hydroxyanisole, butylated hydroxyphenol, butyl hydroxyanisole, hydroxycoumarin, butylated hydroxytoluene, cephalm, ethyl gallate, propyl gallate, octyl gallate, lauryl gallate, propyl hydroxybenzoate, trihydroxybutyrophenone, dimethylphenol, ditert butylphenol, vitamin E, and lecithin. In some embodiments, the formulation comprises SAIB.

[0155] In some embodiments, the formulation further comprises a pharmaceutically acceptable carrier or excipient. In some embodiments, the formulation comprises SAIB.

[0156] In some embodiments, the formulation releases at least about 1% to about 20% of the modified Cav-1 peptide within 4 hours under physiological conditions. In some embodiments, the formulation releases at least about 1% to about 20% of the modified Cav-1 peptide within 4 hours when tested in an elution device at neutral pH. In some embodiments, the formulation releases at least about 1% to about 20% of the modified Cav-1 peptide within 4 hours when tested in an elution device at about 37°C. In some embodiments, the formulation releases at least about 2% to about 15% (including any value or sub-range therebetween) within 4 hours under physiological conditions. In some embodiments, the formulation releases at least about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, or about 15% of the peptide within 4 hours under physiological conditions. In some embodiments, physiological conditions are in an aqueous buffer in the range of about pH 6 to about pH 8 at about 37°C. In some embodiments, physiological conditions are in an aqueous buffer at about pH 7.4 at about 37°C. In some embodiments, the formulation comprises SAIB.

[0157] In some embodiments, the formulation releases the modified Cav-1 peptide under physiological conditions for at least 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, or 42 days, or over any value in between. In some embodiments, the formulation releases the modified Cav-1 peptide under physiological conditions for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, or 12 weeks, or over any value or range in between. In some embodiments, the physiological conditions are in an aqueous buffer at about 37°C in the range of about pH 6 to about pH 8. In some embodiments, the physiological conditions are in an aqueous buffer at about 37°C and about pH 7.4.

[0158] In some embodiments, the formulations described herein release the modified Cav-1 peptide for at least 8 hours under physiological conditions. In some embodiments, the formulations described herein release the modified Cav-1 peptide for at least 12 hours under physiological conditions. In some embodiments, the formulations described herein release the modified Cav-1 peptide for at least 1 day under physiological conditions. In some embodiments, the formulations described herein release the modified Cav-1 peptide for at least 2 days under physiological conditions. In some embodiments, the formulation releases the modified Cav-1 peptide for at least 5 days under physiological conditions. In some embodiments, the formulation releases the modified Cav-1 peptide for at least 7 days under physiological conditions. In some embodiments, the formulation releases the modified Cav-1 peptide for at least 14 days under physiological conditions. In some embodiments, the formulation releases the modified Cav-1 peptide for at least 21 days under physiological conditions. In some embodiments, the formulation releases the modified Cav-1 peptide for at least 28 days under physiological conditions. In some embodiments, the formulation releases the modified Cav-1 peptide for at least 35 days under physiological conditions. In some embodiments, the formulation releases the modified Cav-1 peptide for at least 42 days under physiological conditions. In some embodiments, the release of the modified Cav-1 peptide under physiological conditions is measured by HPLC after placing the formulation in a medium (such as PBS) that mimics physiological conditions. In some embodiments, the formulation is placed in a medium that mimics physiological conditions while being stirred or vibrated. In some embodiments, the physiological conditions are in an aqueous buffer at about 37 °C in the range of about pH 6 to about pH 8. In some embodiments, the physiological conditions are in an aqueous buffer at about 37 °C and about pH 7.4.

[0159] In some embodiments, the single dose of the formulation described herein releases the modified Cav-1 peptide at an average of about mg (1 mg / kg) to about 3 mg / kg of the peptide per kg of the subject's body weight per day under physiological conditions for at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days, or any value therebetween. In some embodiments, the single dose of the formulation releases the modified Cav-1 peptide at an average of about 1 mg / kg to about 3 mg / kg per day under physiological conditions for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, or any value therebetween. In some embodiments, the physiological conditions are in an aqueous buffer at about 37°C in the range of about pH 6 to about pH 8. In some embodiments, the physiological conditions are in an aqueous buffer at about 37°C and about pH 7.4. In some embodiments, the formulation comprises SAIB.

[0160] In some embodiments, the single dose of the formulation described herein releases the modified Cav-1 peptide at an average of about 1 mg (1 mg / kg per day) to about 3 mg / kg of the peptide per kg of the subject's body weight under physiological conditions for at least 2 days. In some embodiments, the single dose of the formulation releases the modified Cav-1 peptide at an average of about 1 mg / kg to about 3 mg / kg per day under physiological conditions for at least 5 days. In some embodiments, the single dose of the formulation releases the modified Cav-1 peptide at an average of about 1 mg / kg to about 3 mg / kg per day under physiological conditions for at least 7 days. In some embodiments, the single dose of the formulation releases the modified Cav-1 peptide at an average of about 1 mg / kg to about 3 mg / kg per day under physiological conditions for at least 14 days. In some embodiments, the single dose of the formulation releases the modified Cav-1 peptide at an average of about 1 mg / kg to about 3 mg / kg per day under physiological conditions for at least 21 days. In some embodiments, the single dose of the formulation releases the modified Cav-1 peptide at an average of about 1 mg / kg to about 3 mg / kg per day under physiological conditions for at least 28 days. In some embodiments, the single dose of the formulation releases the modified Cav-1 peptide at an average of about 1 mg / kg to about 3 mg / kg per day under physiological conditions for at least 35 days. In some embodiments, the single dose of the formulation releases the modified Cav-1 peptide at an average of about 1 mg / kg to about 3 mg / kg per day under physiological conditions for at least 42 days. In some embodiments, the physiological conditions are in an aqueous buffer at about 37°C and in the range of about pH 6 to about pH 8. In some embodiments, the physiological conditions are in an aqueous buffer at about 37°C and at about pH 7.4.

[0161] In some embodiments, the single dose of the formulations described herein releases the modified Cav-1 peptide at an average of about 1 mg (1 mg / kg), about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, or about 3 mg / kg of the peptide per kg of the subject's body weight under physiological conditions over at least 2 days. In some embodiments, the single dose of the formulations described herein releases the modified Cav-1 peptide at an average of about 1 mg (1 mg / kg), about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, or about 3 mg / kg of the peptide per kg of the subject's body weight under physiological conditions over at least 7 days, at least 14 days, at least 21 days, at least 28 days, at least 35 days, or at least 42 days. In some embodiments, the physiological conditions are in an aqueous buffer at about 37°C in the range of about pH 6 to about pH 8. In some embodiments, the physiological conditions are in an aqueous buffer at about 37°C and about pH 7.4.

[0162] IV. Method of Use of the Formulation for Treatment In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is used to treat or prevent a fibrotic disease or disorder. In some embodiments, the modified Cav-1 peptide is a fibrotic disease or disorder such as interstitial lung disease, liver fibrosis, kidney fibrosis, skin fibrosis, glomerulonephritis, systemic sclerosis, cardiac fibrosis, myocardial fibrosis, kidney fibrosis, cirrhosis, nephrosclerosis, arteriosclerosis, macular degeneration, ocular scarring, cataract, retinopathy and vitreoretinopathy, Graves ophthalmopathy, neurofibromatosis, scleroderma, glioblastoma, keloid and hypertrophic scarring, peritoneal fibrotic disease, chronic obstructive pulmonary disease, postoperative myoma, diabetic nephropathy, gynecological cancer, myeloproliferative syndrome, myeloid leukemia, myelodysplastic syndrome, inflammatory bowel disease, non-alcoholic fatty liver disease, fibrosarcoma, rheumatoid arthritis, non-alcoholic steatohepatitis, Alport syndrome, or chronic COVID syndrome is used to treat or prevent.

[0163] In some embodiments, the present disclosure provides a method of treating or preventing a fibrotic 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 formulation thereof. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is used to treat or prevent a lung disease or disorder in a subject. In some embodiments, the modified Cav-1 peptide is, 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 pneumonia, bronchiolitis, sarcoidosis, scleroderma, or lung infection, etc. is used to treat or prevent a lung disease or disorder. In some embodiments, the modified Cav-1 peptide is used to treat or prevent a lung disease or disorder in an elderly or aged subject. In some embodiments, the elderly subject has an interstitial lung disease, such as idiopathic pulmonary fibrosis.

[0164] In some embodiments, the modified Cav-1 peptides or formulations of the present disclosure are used to treat or prevent pulmonary infections in a subject, such as bacterial, viral, or fungal infections. In some embodiments, the pulmonary infection causes in the subject one or more pulmonary diseases or disorders including, but not limited to, ALI, ARDS, COPD, asthma, interstitial lung disease, pulmonary fibrosis, pneumonia, hypersensitivity pneumonia, bronchiolitis, sarcoidosis, and scleroderma.

[0165] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations 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, Chlamydophilia pneumoniae, Chlamydophilia psittaci, Coxiella burnetti, Salmenellosis, Yersina pestis, Mycobacterium leprae, Mycobacterium africanum, Mycobacterium asiaticum, Mycobacterium aviuin-intracellulaire, Mycobacterium chelonei, Mycobacterium abscessus, Mycobacterium fallax, Mycobacterium fortuitum, Mycobacterium kansasii, Mycobacterium leprae, Mycobacterium malmoense, Mycobacterium shimoidei, Mycobacterium simiae, Mycobacterium szulgai, Mycobacterium xenopi, Mycobacterium tuberculosis, Brucella melitensis, Brucella suis, Brucella abortus, Brucella canis, Legionella pneumonophilia, Francisella tularensis, Pneurnocystis carinii, Mycoplasma pneumoniae, or Burkholderia cepacia. In some embodiments, the bacterial infection causes pneumonia in the subject.

[0166] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent viral infections in a subject. In some embodiments, the modified Cav-1 peptides are used to treat or prevent infections in a subject caused by double-stranded DNA (dsDNA) viruses, single-stranded DNA (ssDNA) viruses, single-stranded RNA (ssRNA) viruses, or double-stranded RNA (dsRNA) viruses. 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, torquetenovirus, 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.

[0167] In some embodiments, SARS-CoV-1 causes severe acute respiratory syndrome (SARS) in a subject. In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent SARS in a subject. SARS is initially characterized by systemic symptoms of myalgia, headache, and fever, followed by the onset of respiratory symptoms, mainly cough, dyspnea, and pneumonia, within 2 to 14 days.

[0168] In some embodiments, MERS-CoV causes Middle East respiratory syndrome (MERS) in a subject. In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is 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, especially in individuals with underlying diseases, leading to death. There is no specific drug treatment for MERS, and infection prevention and control measures are essential to prevent its spread in healthcare facilities. See Zumla et al. Lancet 2015;386(9997):995-1007.

[0169] In some embodiments, SARS-CoV-2 causes coronavirus disease 2019 (COVID-19) in a subject. In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation 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 or pharmaceutical formulation is used to treat or prevent an infection caused by a variant of SARS-CoV-2. In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent an infection caused by the SARS-CoV-2 alpha strain. In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent an infection caused by the SARS-CoV-2 beta strain. In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent an infection caused by the SARS-CoV-2 gamma strain. In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent an infection caused by the SARS-CoV-2 delta strain. In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent an infection caused by the SARS-CoV-2 epsilon strain. In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent an infection caused by the SARS-CoV-2 zeta strain. In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent an infection caused by the SARS-CoV-2 eta strain. In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent an infection caused by the SARS-CoV-2 theta strain.In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent an infection caused by the SARS-CoV-2 iota strain. In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent an infection caused by the SARS-CoV-2 kappa strain. In some embodiments, the modified Cav-1 peptide is used to treat or prevent post-acute COVID-19 symptoms caused by the SARS-CoV-2 lambda strain. In some embodiments, the modified Cav-1 peptide is used to treat or prevent post-acute COVID-19 symptoms caused by the SARS-CoV-2 mu strain. In some embodiments, the modified Cav-1 peptide is used to treat or prevent post-acute COVID-19 symptoms caused by the SARS-CoV-2 omicron strain. 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 a sub-strain thereof, or a recombinant thereof. See Konings et al., Variants of Interest and Concern naming scheme conducive for global discourse. Nature Microbiology (2021). In some embodiments, the sub-strain 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 sub-strains of the SARS-CoV-2 variant B.1.1.7 are Q.1, Q.2, Q.3, Q.4, Q.5, Q.6, Q.7, or Q.8. In some embodiments, the sub-strains of the SARS-CoV-2 variant B.1.351 are 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-strains of the SARS-CoV-2 variant P.1 are 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-strains of the SARS-CoV-2 variant B.1.617 are B.1.617.1, B.1.617.2, or B.1.617.3. In some embodiments, the sub-strain of the SARS-CoV-2 variant B.1.526 is B.1.526.1. In some embodiments, the sub-strains of the SARS-CoV-2 variant B.1.621 are B.1.621.1, B.1.621.2, BB.1, or BB.2. In some embodiments, the sub-strain of the SARS-CoV-2 variant C.37 is C.37.1.

[0170] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent fungal infections in a subject. Examples of fungi that cause lung infections include, but are not limited to, Candida (e.g., Candida albicans, Candida glabrata, Candida krusei), Aspergillus, Pneumocystis, Coccidioides (e.g., Coccidioides immitis, Coccidioides posadasii), Blastomyces (e.g., Blastomyces 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, the fungal infection causes invasive pulmonary aspergillosis in the subject. In some embodiments, the fungal infection causes allergic asthma, allergic bronchopulmonary aspergillosis, or hypersensitivity pneumonitis in the subject. In some embodiments, the fungal infection causes ARDS. In some embodiments, the fungal infection causes pulmonary fibrosis in the subject. In some embodiments, the fungal infection causes pulmonary edema in the subject.

[0171] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent interstitial lung disease in a subject. Interstitial lung disease is a group of disorders that cause fibrosis and inflammation of the interstitium. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis, lymphangioleiomyomatosis, nonspecific interstitial pneumonia, idiopathic interstitial pneumonia, idiopathic 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.

[0172] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent acute lung injury (ALI) in a subject. ALI is an acute inflammatory disorder that causes disruption of the pulmonary endothelial and epithelial barriers. ALI can be the result of an inhalational injury, or 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.

[0173] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent ARDS in a subject. ARDS is the most severe form of ALI and is distinguished by the severity of hypoxemia. ARDS is a life-threatening type of lung injury that results from the accumulation of fluid in the small elastic air sacs (alveoli) of the lungs. The fluid in the alveoli prevents the lungs from being filled with oxygen, resulting in less oxygen reaching the bloodstream and causing difficulty breathing.

[0174] In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent cystic fibrosis (CF) in a subject. CF is a hereditary disease of the exocrine glands and exocrine sweat glands that mainly affects the digestive and respiratory systems. This disease is usually characterized by chronic respiratory infections, pancreatic insufficiency, abnormally viscous mucous secretions, and premature death. CF is characterized by progressive airflow obstruction. A subset of individuals with CF also develop airway hyperresponsiveness to inhaled cholinergic agonists (Weinberger, 2002 and Mitchell et al., 1978) and reversibility of 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 a possible common etiology between CF and other airway stenosis diseases, such as asthma or COPD, where airway smooth muscle dysfunction is thought to contribute to the disease process.

[0175] In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent COPD in a subject. COPD is a term used to classify two major airflow obstruction disorders, chronic bronchitis and emphysema. Chronic bronchitis is an inflammation of the bronchial airways. The bronchial airways connect the trachea to the lungs. When inflammation occurs, the bronchi secrete mucus, causing a chronic cough. In emphysema, alveolar sacs are overinflated as a result of damage to the elastic skeleton of the lungs. Inflammatory cells in the emphysematous lungs release elastase enzymes, which break down or damage the elastic fibers within the lung matrix. Emphysema has several causes, including smoking, exposure to environmental pollutants, alpha1-antitrypsin deficiency, and aging.

[0176] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations disclosed herein are 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 the peripheral airways, and increased mucus production (Ralston et al., 2014). Signs and symptoms typically begin with rhinitis and cough, which can progress to tachypnea, wheezing, rales, use of accessory muscles, and / or nasal flaring.

[0177] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations disclosed herein are used to treat or prevent obliterative bronchiolitis in a subject. Obliterative bronchiolitis is a progressive airflow limitation as a result of abnormal remodeling of the peripheral airways of the lung (Meyer et al., 2014). Obliterative bronchiolitis is a major complication of lung transplantation and is often used to explain late-onset allograft dysfunction, which results in a persistent decline in forced expiratory volume and expiratory force that is not caused by other known causes (Meyer et al., 2014).

[0178] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations disclosed herein are used to treat or prevent asthma in a subject. 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, asymptomatic asthma, gastroesophageal asthma, idiopathic asthma, and cough variant asthma. During an asthma attack, the airways become persistently inflamed and may sometimes spasm.

[0179] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations disclosed herein are used to treat or prevent hypersensitivity pneumonitis in a subject. Hypersensitivity pneumonitis is a complex syndrome that is susceptibility-prone and caused by the inhalation of various antigens in sensitized individuals. These antigens are found in the environment and mostly originate from avian proteins and fungi. Hypersensitivity pneumonitis is characterized by excessive humoral and cellular immune responses that affect the peripheral airways and lung parenchyma. Hypersensitivity pneumonitis can be classified into acute, chronic non-fibrotic, and chronic fibrotic forms. Acute hypersensitivity pneumonitis occurs usually within hours of exposure from intermittent high-level exposure to the inducing antigen, while chronic hypersensitivity pneumonitis is mainly due to long-term low-level exposure (usually to birds or molds in the home), which is not easily defined in terms of time and can occur within weeks, months, or even years from exposure. Some patients with fibrotic hypersensitivity pneumonitis may progress to a progressive phenotype even if exposure is completely avoided. See Costabel et al., Nature Reviews Disease Primers 2020;6(65).

[0180] In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent systemic sclerosis or scleroderma in a subject. Systemic sclerosis is a systemic autoimmune disease characterized by endothelial dysfunction leading to small vessel vasculopathy, resulting fibroblast dysfunction with excessive collagen production and fibrosis, and immunological abnormalities. The classification of systemic sclerosis is subdivided, based on the extent of skin lesions, into diffuse cutaneous systemic sclerosis, limited cutaneous systemic sclerosis, or systemic sclerosis without skin sclerosis. Virtually every organ system can be involved in the disease process, but the fibrotic and vascular lung manifestations of systemic sclerosis, including interstitial lung disease and pulmonary hypertension, are the major causes of death. Certain lung manifestations may occur more commonly in subsets of systemic sclerosis (i.e., ILD is more common in diffuse cutaneous systemic sclerosis while pulmonary hypertension is more common in limited cutaneous systemic sclerosis), but all of the known lung manifestations that have been reported are described in each of the subsets of the disease. Lung disease can also occur in systemic sclerosis without skin lesions (an element known as scleroderma sine scleroderma). See Solomon et al., Eur Respir Rev 2013;22(127):6-19.

[0181] In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent sarcoidosis in a subject. Sarcoidosis is a multi-system disorder characterized by non-caseating epithelioid cell granulomas that can affect almost all organs. Chest lesions are common and account for most of the morbidity and mortality associated with this disease. Chest abnormalities are observed in approximately 90% of patients with sarcoidosis, and an estimated 20% develop chronic lung disease leading to pulmonary fibrosis. Pulmonary sarcoidosis can present in various patterns. Bilateral hilar lymphadenopathy is the most common finding, followed by interstitial lung disease. The most typical findings of lung lesions are small nodules with a perilymphatic distribution, fibrotic changes, and bilateral perihilar shadows. Atypical features such as mass or alveolar shadows, honeycombed cysts, miliary shadows, mosaic opacities, tracheobronchial lesions, and pleural disease, as well as complications such as aspergilloma, may also be seen. See Criado et al., Chest Imaging 2010;30(6).

[0182] In some embodiments, the present disclosure provides a method of treating or preventing a lung 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 formulation thereof.

[0183] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is used to treat or prevent a disease or disorder of the kidney (e.g., chronic kidney disease) in a subject. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is used to treat or prevent a disease or disorder of the lung (e.g., idiopathic pulmonary fibrosis) in a subject. In some embodiments, the subject is elderly or aged.

[0184] In some embodiments, the modified Cav-1 peptide formulation is used to treat or prevent kidney diseases or disorders 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 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 or aged. In some embodiments, the kidney disease or disorder in the subject is caused by an infectious disease such as, for example, a viral infection, a bacterial infection, a fungal infection, or a parasitic infection. In some embodiments, the kidney disease or disorder in the subject is caused by hypertension (hypertensive disease). 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 the excessive use of drugs such as commercially available analgesics and heroin. In some embodiments, the subject has hypertensive disease or diabetes.

[0185] In some embodiments, the modified Cav-1 peptide or its pharmaceutical formulation is used to delay the progression of a kidney disease or disorder in a subject. In some embodiments, the modified Cav-1 peptide or its pharmaceutical formulation is used to improve the progression-free survival period. In some embodiments, the modified Cav-1 peptide or its pharmaceutical formulation is used to extend the survival period of a subject.

[0186] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent chronic kidney disease in a subject. Chronic kidney disease is the progressive and gradual loss of the kidney's ability to excrete waste, concentrate urine, and conserve electrolytes. The progressive loss of kidney function results from the deposition of fibrous tissue (interstitial fibrosis) between the kidney or the functional units of the nephron, and the continuous replacement of the filtration surface by fibrous tissue (glomerulosclerosis). Renal fibrosis is a pathological feature of chronic kidney disease and a major contributing factor to the progression to end-stage renal disease. In one embodiment, the chronic kidney disease is chronic renal fibrosis.

[0187] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent end-stage renal disease in a subject. End-stage renal disease is the final stage of chronic kidney disease in which the kidneys have ceased to function, and the individual requires long-term dialysis or a kidney transplant to survive.

[0188] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations 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 that causes loss of protein in the urine.

[0189] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent glomerulonephritis in a subject. Glomerulonephritis, also referred to as glomerular disease, is a type of kidney disease in which the 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.

[0190] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are 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 within the kidneys, causing the kidneys to enlarge over time and lose function.

[0191] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent IgA nephropathy in a subject. IgA nephropathy, also known as Berger's disease, is a kidney disease that occurs when the immune globulin IgA accumulates in the kidneys, causing local inflammation, which can interfere with the kidneys' ability to filter waste products from the blood.

[0192] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent lupus nephritis in a subject. Lupus nephritis is a type of glomerulonephritis that constitutes one of the most severe organ manifestations of systemic lupus erythematosus and occurs when the immune system attacks the kidneys.

[0193] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent nephrotic syndrome in a subject. Nephrotic syndrome is a kidney disorder in which the body excretes excessive amounts of protein in the urine. Nephrotic syndrome is often caused by damage to the tiny blood vessels in the kidneys that filter waste products and excess fluid from the blood.

[0194] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent Alport syndrome in a subject. Alport syndrome is a genetic disorder characterized by progressive kidney disease and abnormalities of the inner ear and eyes. There are three genotypes: X-linked Alport syndrome (XLAS), autosomal recessive Alport syndrome (ARAS), and autosomal dominant Alport syndrome (ADAS). XLAS is caused by variants in the COL4A5 gene, ARAS is caused by variants in both copies of either the COL4A3 or COL4A4 gene, and ADAS is caused by variants in one copy of the COL4A3 or COL4A4 gene. Individuals with Alport syndrome present with chronic glomerular dysfunction, kidney inflammation, and fibrosis characteristic of chronic kidney disease and progress to end-stage kidney disease. Individuals affected by Alport syndrome may also develop progressive hearing loss of varying severity and eye abnormalities that usually do not result in visual impairment.

[0195] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent amyloidosis in a subject. Amyloidosis occurs when amyloid accumulates in tissues and organs, interfering with normal function. Amyloid deposits damage the kidneys, affecting their ability to filter waste products and break down proteins. In some embodiments, the amyloidosis is primary amyloidosis. In some embodiments, the amyloidosis is dialysis-related amyloidosis.

[0196] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent Goodpasture syndrome in a subject. Goodpasture syndrome, also known as anti-glomerular basement membrane disease, is an autoimmune disease in which antibodies attack the basement membranes of the lungs and kidneys, resulting in pulmonary hemorrhage, glomerulonephritis, and renal failure.

[0197] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations 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 associated with granulomatous inflammation, necrosis, and vasculitis, most frequently targeting the lungs and kidneys.

[0198] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations of the present disclosure are used to treat or prevent acute kidney injury in a subject. Acute kidney injury, also known as acute renal failure, is a sudden loss of renal excretory function. Acute kidney injury is defined by serum creatinine and urine output levels over a period of less than one week.

[0199] In some embodiments, the modified Cav-1 peptides or pharmaceutical formulations are used to treat or prevent kidney infections. In some embodiments, the modified Cav-1 peptides are 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, for example, Escherichia coli, Klebsiella, Proteus, Pseudomonas, Enterococcus, or Staphylococcus saprophyticus.

[0200] In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent kidney diseases or disorders resulting from microbial infections in a subject. In some embodiments, the microbial infection is a bacterial infection, a viral infection, a fungal infection, or a parasitic infection. In some embodiments, the kidney disease or disorder is caused by Streptococcus pyogenes, Staphylococcus (aureus, epidermidis), Salmonella (typhi, paratyphi), Escherichia coli, Leptospira, Mycobacterium tuberculosis, Mycobacterium leprae, Legionella spp., Yersinia enterocolitica, Brucella spp., Campylobacter jejuni, Corynebacterium diphtheriae, Klebsiella, Proteus, Pseudomonas, Enterococcus, 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.

[0201] In some embodiments, the modified Cav-1 peptide or pharmaceutical formulation is used to treat or prevent kidney diseases or disorders 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.

[0202] In some embodiments, the kidney disease or kidney disorder is a chronic kidney disease. In some embodiments, the kidney disease or kidney disorder is Alport syndrome. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is used to improve kidney function in a subject having a kidney disease or kidney disorder. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof improves kidney 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% as compared to the subject prior to treatment with the modified Cav-1 peptide or a pharmaceutical formulation thereof. In some embodiments, the improvement in kidney function is represented by a decrease in fibrotic glomeruli, a decrease in blood urea nitrogen, a decrease in serum creatinine, an increase in blood albumin, a decrease in the urinary albumin / creatinine ratio, and / or an increase in glomerular filtration rate.

[0203] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof decreases the number of fibrotic glomeruli in a subject having a kidney disease or kidney disorder. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof decreases 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% as compared to the subject prior to treatment with the modified Cav-1 peptide or a pharmaceutical formulation thereof.

[0204] In some embodiments, a subject having a kidney disease or disorder has a reduced caveolin-1 expression in the kidney compared to a subject without a kidney disease or disorder (e.g., a normal healthy subject). In some embodiments, the caveolin-1 expression is reduced in the glomeruli of a subject having a kidney disease or disorder compared to a subject without a kidney disease or disorder. In some embodiments, the caveolin-1 expression is reduced in the endothelial cells of the kidney of a subject having a kidney disease or disorder compared to a subject without a kidney disease or disorder. In some embodiments, the caveolin-1 expression is reduced in the epithelial cells of the kidney of a subject having a kidney disease or disorder compared to a subject without a kidney disease or disorder. In some embodiments, the caveolin-1 expression is reduced in the podocytes of a subject having a kidney disease or disorder compared to a subject without a kidney disease or disorder in the kidney. In some embodiments, the caveolin-1 expression 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 having a kidney disease or disorder compared to a subject without a kidney disease or disorder. In some embodiments, the epithelial cells are parietal epithelial cells covering the Bowman's capsule.

[0205] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof reduces endothelial cell death in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof reduces epithelial cell death in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof reduces podocyte cell death in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation 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 a pharmaceutical formulation thereof.

[0206] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof increases endothelial cell survival in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof increases epithelial cell survival in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof increases podocyte survival in a subject having a kidney disease or disorder. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof increases cell survival 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 a pharmaceutical formulation thereof.

[0207] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof promotes kidney regeneration in a subject having a kidney disease or kidney disorder. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof promotes the regeneration of renal blood vessels, glomeruli, and / or tubules in the subject's kidney. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof promotes the regeneration of epithelial cells, endothelial cells, tubular cells, and / or podocytes in the subject's kidney.

[0208] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof increases endothelial cell proliferation in a subject having a kidney disease or kidney disorder. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof increases epithelial cell proliferation in a subject having a kidney disease or kidney disorder. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof increases podocyte proliferation in a subject having a kidney disease or kidney disorder. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation 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 before treatment with the modified Cav-1 peptide or a pharmaceutical formulation thereof.

[0209] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof reduces blood urea nitrogen in a subject having a kidney disease or disorder. In some embodiments, a high blood urea nitrogen value indicates kidney injury or disease in the subject. Generally, in humans, blood urea nitrogen levels in the range of 6 mg / dl to 24 mg / dl are considered normal. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation 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 prior to treatment with the modified Cav-1 peptide or a pharmaceutical formulation thereof. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof reduces blood urea nitrogen in the 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 a pharmaceutical formulation thereof reduces blood urea nitrogen to less than about 20 mg / dl.

[0210] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof reduces blood creatinine in a subject having a kidney disease or disorder. In some embodiments, a high blood creatinine value indicates kidney injury or disease in the subject. Generally, a blood creatinine value exceeding 1.2 mg / dl in females and 1.4 mg / dl in males indicates that the kidneys are not functioning properly. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation 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 prior to treatment with the modified Cav-1 peptide or a pharmaceutical formulation thereof. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation 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 a pharmaceutical formulation thereof reduces blood urea nitrogen to less than about 1.5 mg / dl.

[0211] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof increases blood albumin in a subject having a kidney disease or disorder. In some embodiments, a low blood albumin level may indicate kidney injury or disease in a subject. The normal level of albumin in the blood is 3.5 g / dL to 5 g / dL. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation 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 a pharmaceutical formulation thereof. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof increases blood albumin in the subject to more than about 1.5 g / dl, more than about 1.75 g / dl, more than about 2 g / dl, more than about 2.25 g / dl, more than about 2.5 g / dl, more than about 2.75 g / dl, more than about 3 g / dl, or more than about 3.5 g / dl. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof increases blood albumin to more than about 3.5 g / dl.

[0212] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof reduces the urinary albumin / creatinine ratio in a subject having a kidney disease or disorder. The urinary albumin / creatinine ratio is useful for identifying kidney injury or disease in a subject. In humans, a ratio of albumin to creatinine of less than 30 mg / g is considered normal, a ratio of 30 - 300 mg / g represents microalbuminuria, and values above 300 mg / g represent overt albuminuria. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof reduces the urinary albumin / 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% as compared to the subject prior to treatment with the modified Cav-1 peptide or a pharmaceutical formulation thereof. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof reduces the urinary albumin / creatinine ratio 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 / g in the subject. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof reduces the urinary albumin / creatinine ratio to less than about 30 mg / g.

[0213] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof increases the glomerular filtration rate in a subject having a kidney disease or disorder. The glomerular filtration rate measures how well the kidneys filter blood to remove waste products and excess water to form urine. A glomerular filtration rate above 90 mL / min / 1.73 m 2 is considered normal, while a glomerular filtration rate less than 60 mL / min / 1.73 m 2 may indicate kidney injury or disease. A glomerular filtration rate of 15 mL / min / 1.73 m 2Low glomerular filtration rate may indicate renal failure. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation 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 before treatment with the modified Cav-1 peptide or a pharmaceutical formulation thereof. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof increases the glomerular filtration rate in a subject to greater than about 30 mL / min / 1.73m 2 sup, greater than about 40 mL / min / 1.73m 2 sup, greater than about 50 mL / min / 1.73m 2 sup, greater than about 60 mL / min / 1.73m 2 sup, greater than about 70 mL / min / 1.73m 2 sup, greater than about 80 mL / min / 1.73m 2 sup, or greater than about 90 mL / min / 1.73m 2 sup. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof increases the glomerular filtration rate in a subject to greater than about 60 mL / min / 1.73m 2 sup.

[0214] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is used to preserve renal function in a subject having a renal disease or disorder. As used herein, the term "preserve" refers to maintaining renal function or preventing further decline in renal function in a subject having a renal disease or disorder. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof preserves renal function such that these measurements remain stable after treatment with the modified Cav-1 peptide or a pharmaceutical formulation thereof, as measured by blood urine nitrogen, serum creatinine, serum albumin, urine albumin / creatinine ratio, and / or glomerular filtration rate.

[0215] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is used for treating or preventing a disease or disorder in an elderly subject. As used herein, the term "elderly" or "aged" refers to a subject 55 years of age 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 has an increased susceptibility to the diseases or disorders described herein as compared to a younger subject. In some embodiments, the elderly subject has a fibrotic disease or disorder, such as idiopathic pulmonary fibrosis.

[0216] In some embodiments, the elderly subject has a decreased caveolin-1 expression as compared to a younger subject (e.g., a young adult or middle-aged subject). In some embodiments, the caveolin-1 expression is decreased 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 the elderly subject as compared to the younger subject.

[0217] 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 formulation thereof.

[0218] In some embodiments of any one of the methods disclosed herein, a formulation comprising a modified Cav-1 peptide disclosed herein comprises a modified Cav-1 peptide comprising any one of the amino acid sequences of SEQ ID NOs: 2 to 111. In some embodiments of any one of the methods disclosed herein, a formulation comprising a modified Cav-1 peptide disclosed herein comprises a modified Cav-1 peptide comprising any one of the amino acid sequences of SEQ ID NOs: 2 to 10. In some embodiments of any one of the methods disclosed herein, a formulation comprising a modified Cav-1 peptide disclosed herein comprises a modified Cav-1 peptide comprising the amino acid sequence of SEQ ID NO: 3. In some embodiments of any one of the methods disclosed herein, a formulation comprising a modified Cav-1 peptide disclosed herein comprises a modified Cav-1 peptide comprising the amino acid sequence of SEQ ID NO: 8. In some embodiments of any one of the methods disclosed herein, a formulation comprising a modified Cav-1 peptide disclosed herein comprises a modified Cav-1 peptide comprising at least one amino acid substitution, deletion or insertion relative to the amino acid sequence of FTTFTVT (SEQ ID NO: 3), and the modified Cav-1 peptide maintains the biological activity of Cav-1.

[0219] In some embodiments of any one of the methods disclosed herein, a formulation comprising a modified Cav-1 peptide disclosed herein comprises an effective amount of a modified Cav-1 peptide comprising any one of the amino acid sequences of SEQ ID NOs: 2 to 111. In some embodiments of any one of the methods disclosed herein, a formulation comprising a modified Cav-1 peptide disclosed herein comprises an effective amount of a modified Cav-1 peptide comprising any one of the amino acid sequences of SEQ ID NOs: 2 to 10. In some embodiments of any one of the methods disclosed herein, a formulation comprising a modified Cav-1 peptide disclosed herein comprises an effective amount of a modified Cav-1 peptide comprising the amino acid sequence of SEQ ID NO: 3. In some embodiments of any one of the methods disclosed herein, a formulation comprising a modified Cav-1 peptide disclosed herein comprises an effective amount of a modified Cav-1 peptide comprising the amino acid sequence of SEQ ID NO: 8. In some embodiments of any one of the methods disclosed herein, a formulation comprising a modified Cav-1 peptide disclosed herein comprises an effective amount of a modified Cav-1 peptide comprising at least one amino acid substitution, deletion, or insertion relative to the amino acid sequence of FTTFTVT (SEQ ID NO: 3), and the modified Cav-1 peptide maintains the biological activity of Cav-1.

[0220] The present disclosure contemplates all modes of administration, dosages, or dosing frequencies suitable for treating or preventing a disease or disorder in a subject. Effective dosages may also be extrapolated from dose-response curves derived from in vitro or animal model test bioassays or systems.

[0221] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered systemically. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered intravenously, intrathecally, subcutaneously, and / or intraperitoneally.

[0222] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is locally delivered to the airways of a subject, such as by administration of a nebulized formulation using a nebulizer or a dry powder formulation using a dry powder inhaler. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to the lungs of a subject using a nebulizer. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to the lungs of a subject using a dry powder inhaler. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject intranasally, intratracheally, intrapleurally, intratracheally, or via inhalation.

[0223] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject as a single dose or multiple doses. When multiple doses are administered, the doses can be spaced apart from each other by, for example, 1 hour, 3 hours, 6 hours, 8 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 1 year, or any value or range therebetween. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered, for example, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 5 weeks, once every 6 weeks, once every 7 weeks, once every 8 weeks, once every 10 weeks, once every 15 weeks, once every 20 weeks, or once at a longer interval. It should be understood that for any particular subject, the specific dosing regimen should be adjusted over time according to the individual needs and the professional judgment of the person administering or overseeing the administration of the formulation. For example, the dosage of the modified Cav-1 peptide or a pharmaceutical formulation thereof can be increased if sufficient therapeutic activity is not obtained at a lower dosage.

[0224] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a 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 a pharmaceutical formulation thereof is administered to a 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 a pharmaceutical formulation thereof is administered to a subject at a dose of about 0.01 mg / kg to about 1 mg / kg. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 1 mg / kg to about 100 mg / kg. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 1 mg / kg to about 50 mg / kg. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 1 mg / kg to about 25 mg / kg. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 1 mg / kg to about 10 mg / kg. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 10 mg / kg to about 25 mg / kg. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 25 mg / kg to about 50 mg / kg. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 50 mg / kg to about 75 mg / kg. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 75 mg / kg to about 100 mg / kg. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation 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. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 1 mg / kg to about 10 mg / kg.In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 2 mg / kg to 5 mg / kg. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, or any value or range therebetween. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 1 mg / kg to about 5 mg / kg. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 1.0 mg / kg, about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 4.0 mg / kg, about 4.1 mg / kg, about 4.2 mg / kg, about 4.3 mg / kg, about 4.4 mg / kg, about 4.5 mg / kg, about 4.6 mg / kg, about 4.7 mg / kg, about 4.8 mg / kg, about 4.9 mg / kg, about 5.0 mg / kg, or any value or range therebetween. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered to a subject at a dose of about 2.2 mg / kg.

[0225] In some embodiments, the total or full dose of the modified Cav-1 peptide or its pharmaceutical formulation administered to a subject is from about 1 mg to about 100 mg, such as, for example, from about 20 mg to about 100 mg, from about 50 mg to about 100 mg, from about 10 mg to about 20 mg, from about 20 mg to about 40 mg, from about 50 mg to about 70 mg, or from about 80 mg to about 90 mg. In some embodiments, the total or full dose of the modified Cav-1 peptide or its pharmaceutical formulation administered to a subject is about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 12 mg, about 14 mg, about 16 mg, about 18 mg, about 20 mg, about 22 mg, about 24 mg, about 26 mg, about 28 mg, about 30 mg, about 32 mg, about 34 mg, about 36 mg, about 38 mg, about 40 mg, about 42 mg, about 44 mg, about 46 mg, about 48 mg, about 50 mg, about 52 mg, about 54 mg, about 56 mg, about 58 mg, about 60 mg, about 62 mg, about 64 mg, about 66 mg, about 68 mg, about 70 mg, about 72 mg, about 74 mg, about 76 mg, about 78 mg, about 80 mg, about 82 mg, about 84 mg, about 86 mg, about 88 mg, about 90 mg, about 92 mg, about 94 mg, about 96 mg, about 98 mg, about 100 mg, about 110 mg, about 120 mg, or any value or sub-range therebetween.

[0226] In some embodiments, the modified Cav-1 peptide or its pharmaceutical formulation is prepared as a long-term release formulation. As used herein, the term "long-term release" refers to the ability to release a component (i.e., the modified Cav-1 peptide) over a specified period of time. Without wishing to be bound by any theory, it is contemplated that the formulations of the present disclosure provide long-term release of the modified Cav-1 peptide over a specified period of time, such as, for example, but not limited to, about 1 week or more, about 2 weeks or more, about 3 weeks or more, about 4 weeks or more, about 5 weeks or more, about 6 weeks or more, about 7 weeks or more, about 8 weeks or more, about 12 weeks or more, about 16 weeks or more, about 20 weeks or more, about 24 weeks or more, about 28 weeks or more, about 1 month or more, about 2 months or more, about 3 months or more, about 4 months or more, about 5 months or more, about 6 months or more.

[0227] In some embodiments, the dosage of the modified Cav-1 peptide or its pharmaceutical formulation for a particular subject is determined by one of ordinary skill in the art using conventional considerations (e.g., using appropriate conventional pharmacological protocols). For example, a physician may initially prescribe a relatively low dose and then increase the dose until an appropriate response is obtained. The dose administered to the subject is sufficient to produce a beneficial therapeutic response in the subject over time, depending on the use, or, for example, to reduce symptoms or other appropriate activities. The dose 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, and the condition of the subject, as well as the weight or surface area of the subject being treated.

[0228] In some embodiments, a subject is administered a dose of the modified Cav-1 peptide or its pharmaceutical formulation 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 or of advanced age. In some embodiments, a single dose is, for example, about 0.2 mg / kg to about 250 mg / kg, such as about 1 mg / kg to about 10 mg / kg, about 10 mg / kg to about 25 mg / kg, about 25 mg / kg to about 50 mg / kg, about 50 mg / kg to about 75 mg / kg, about 75 mg / kg to about 100 mg / kg, via, for example, pulmonary instillation (e.g., inhalation). Such doses can be administered daily over any period from about 3 days to more than 1 week, or at any frequency, as disclosed herein. Chronic administration of the modified Cav-1 peptide or its pharmaceutical formulation is also possible, although the dose may need to be adjusted downward, as is well understood in the art. However, the foregoing ranges are suggestive, as there are a large number of variable elements in an individual treatment regimen and significant deviations from these preferred values are expected.

[0229] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered continuously to a subject. For example, in the case of continuous administration by a pump system such as an osmotic pump, the total dosage over a period of about 1 to 2 weeks is in the range of about 1 mg / kg to about 1 g / kg, about 20 to about 300 mg / kg, or about 50 to about 200 mg / kg. After such a continuous dosing regimen, the total concentration of the active compound can be in the range of about 0.5 μM to about 50 μM, or about 1 μM to about 10 μM.

[0230] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered on a conventional schedule. As used herein, a conventional schedule refers to a predetermined specified period. A conventional schedule can include the same period or periods of different lengths as long as the schedule is predetermined. For example, a conventional schedule can involve administration once a day, twice a day, once every two days, once every three days, once every four days, once every five days, once every six days, 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 a pharmaceutical formulation thereof is administered twice a day for the first week and then daily for several months. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered once a day. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered less than once a day, for example, every other day, every three days, or once a week.

[0231] In some embodiments, the modified Cav-1 peptide formulation is administered to a subject 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 formulation is administered to a subject having fibrosis for at least about 2 weeks. In some embodiments, the modified Cav-1 peptide formulation is administered to a subject having fibrosis for at least about 4 weeks.

[0232] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is provided in unit dosage forms (e.g., pre-divided dosages) such as in capsules, blisters, or cartridges. In some embodiments, the unit dosage comprises at least 1 mg of the modified Cav-1 peptide formulation per dosage, 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 formulation. In some embodiments, the unit dosage is a modified Cav-1 peptide formulation of about 1 mg to about 10 mg (e.g., about 5 mg). In some embodiments, the unit dosage form does not include the administration or addition of any excipients and is used simply to hold the powder for inhalation (i.e., the capsule, blister, or cartridge is not administered). In some embodiments, a plurality of the unit dosage forms are administered to a subject. For example, in the case of a dry powder inhaler, the modified Cav-1 peptide formulation is provided in unit dosage capsules, and a plurality of unit dosage capsules (e.g., 3 to 4) can be administered to the subject by inhalation. In some embodiments, the modified Cav-1 peptide formulation is administered at high release dosages such as at least about 10 mg, at least about 15 mg, or at least about 20 mg. In some embodiments, the administration of the milled modified Cav-1 peptide formulation results in a high particulate dosage of more than about 5 mg deep into the lungs. In some embodiments, the particulate dosage deep into the lungs is at least about 10 mg or at least about 15 mg. In some embodiments, the particulate dosage is provided from 1, 2, 3, 4, or 5 or more capsules containing the dosage of the peptide of the embodiment. In some embodiments, the particulate dosage 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 release dosage.

[0233] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation 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. Additional therapeutic agents include, but are not limited to, angiotensin-converting enzyme (ACE) inhibitors such as Capoten® (captopril), Vasotec® (enalapril), Monopril® (fosinopril), Prinivil® or Zestril® (lisinopril), or Altace® (ramipril); angiotensin II receptor (ARB) inhibitors such as Edarbi® (azilsartan), Teveten® (eprosartan), Avapro® (irbesartan), Cozaar® (losartan), Benicar® (olmesartan), or Diovan® (valsartan); Farxiga® (dapagliflozin); Aranesp® (darbepoetin alfa); and / or Procrit® or Epogen® (erythropoietin).

[0234] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered in combination with dialysis for the treatment of a kidney disease or disorder in a subject. In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered in combination with dialysis for the treatment of chronic kidney disease. In some embodiments, the dialysis is hemodialysis. In some embodiments, the dialysis is peritoneal dialysis.

[0235] In some embodiments, the modified Cav-1 peptide or a pharmaceutical formulation thereof is administered simultaneously or sequentially in combination with at least one additional therapeutic agent for pulmonary fibrosis. Additional therapeutic agents include, but are not limited to, non-steroidal anti-inflammatory drugs (NSAIDs), steroids, disease-modifying anti-rheumatic drugs (DMARDs), immunosuppressive agents, biologic response modifiers, bronchodilators or anti-fibrotic agents such as pirfenidone (a drug with an anti-fibrotic mechanism of action not fully understood but which may involve blockade of TGF-beta), nintedanib, a broad-spectrum tyrosine kinase blocker, or any other anti-fibrotic agent. Suitable NSAIDs are acetylsalicylic acid, mesalazine, ibuprofen, naproxen, flurbiprofen, fenoprofen, fenbufen, ketoprofen, indoprofen, pirprofen, carprofen, oxaprozin, pranoprofen, miroprofen, thiaprofenic acid, suprofen, alminoprofen, tiaprofenic acid, fluprofen, indomethacin, sulindac, tolmetin, zomepirac, nabumetone, diclofenac, fenclofenac, alclofenac, bromfenac, ibufenac, aceclofenac, acemetacin, fentiazac, clidanac, etodolac, oxypinac, mefenamic acid, meclofenamic acid, flufenamic acid, niflumic acid, tolfenamic acid, diflunisal, flufenisal, piroxicam, tenoxicam, lornoxicam and nimesulide, and pharmaceutically acceptable salts thereof, meloxicam, celecoxib and rofecoxib, which are selective COX2 inhibitors, and pharmaceutically acceptable salts thereof. Suitable steroids are prednisone, prednisolone, methylprednisolone, dexamethasone, budenoside, fludrocortolone and triamcinolone. Suitable DMARDs are sulfasalazine, olsalazine, chloroquine, gold derivatives (aurothioglucose), D-penicillamine, and cytostatic agents such as methotrexate and cyclophosphamide.Suitable immunosuppressants are cyclosporin A and its derivatives, mycophenolate mofetil, FK 506 (also known as tacrolimus and fujimycin), muromonab-CD3 (Orthoclone OKT-3 (registered trademark)), antithymocyte globulin (ATG), 15-deoxyspergualin, mizoribine, misoprostol, rapamycin, leflunomide and azathioprine. Suitable biological response modifiers are interferon β, anti-TNF-α antibody (etanercept), IL-10, anti-CD3 antibody or anti-CD25 antibody. Suitable bronchodilators are ipratropium bromide, oxitropium bromide, tiotropium bromide, epinephrine hydrochloride, salbutamol, terbutaline sulfate, fenoterol bromide, salmeterol and formoterol. In such combinations, each active ingredient can be administered either according to its normal dosage range or at a lower dosage than its normal dosage range. The dosage of the combination of NSAID, steroid, DMARD, immunosuppressant and biological response modifier is from approximately 1 / 50 of the usually recommended minimum dosage to 1 / 1 of the usually recommended dosage, 1 / 20 to 1 / 2, or 1 / 10 to 1 / 5. The dosage usually recommended for the combined drugs should be understood to be, for example, the dosage disclosed in Rote Liste (registered trademark) 2002, Editio Cantor Verlag Aulendorf, Germany, or Physician’s Desk Reference.

[0236] In some embodiments, the modified Cav-1 peptide or its pharmaceutical formulation is administered simultaneously or sequentially in combination with at least one additional therapeutic agent for treating a pathogen or pathogen-mediated lung injury. Additional therapeutic agents include, but are not limited to, chloroquine, hydroxychloroquine, type I interferon, antiviral drugs, antibiotics, remdesivir, favipiravir, lopinavir, ritonavir, nirmalavir, baricitinib, and molnupiravir.

[0237] Hydroxychloroquine is a chemical derivative of chloroquine, characterized by a hydroxyethyl group instead of an ethyl group. Hydroxychloroquine is classified as an effective antimalarial drug and has shown effectiveness in the treatment of systemic lupus erythematosus, as well as rheumatoid arthritis and Sjogren's syndrome. Hydroxychloroquine has long been known to increase the pH of lysosomes in antigen-presenting cells, but its mechanism of action in inflammatory conditions has only recently been elucidated, which involves blocking the activation of toll-like receptors on plasmacytoid dendritic cells. Hydroxychloroquine has shown effectiveness in the treatment of RNA viruses, including hepatitis C. Hydroxychloroquine can be administered at a dose of 600 mg per day.

[0238] Human type I interferons (IFNs) are a large subgroup of interferon proteins that assist in regulating the activity of the immune system. The types in mammals are designated as 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 various viruses, including picornaviruses, chikungunya virus, flaviviruses, 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 with other peptides and proteins, such as immunoglobulins and other cytokines. Interferon-alpha and interferon-beta conjugates can represent, for example, formulations containing interferon-beta conjugated to a non-natural polymer containing a polyalkylene glycol moiety.Preferred interferon compounds include Roferon® (interferon alpha-2a), Intron® (interferon alpha-2b), Alferon® (interferon alpha-n3), Infergen® (interferon alfa-con1), Omniferon® (interferon alpha), interferon alfa-con1, 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 preparation, interferon-alpha as a topical preparation, Roferon® (interferon alpha-2a) analogs, Intron® (interferon alpha-2b) analogs, Alferon® (interferon alpha-n3) analogs, and Infergen® (interferon alfa-con1) analogs, Omniferon® (interferon alpha) analogs, interferon alfa-con1 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, polymerized interferon-beta, dimerized interferon-beta, interferon-beta conjugated to a carrier, interferon-beta as an oral inhalant, interferon-beta as an injectable preparation, interferon-beta as a topical preparation, 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 interferon-alpha or interferon-beta production or mimic the action of interferon-alpha or interferon-beta may also be used. Interferon inducers include tilorone, poly(I)-poly(C), imiquimod, cridanimod, and bropirimine.

[0239] In some embodiments, the modified Cav-1 peptide or its pharmaceutical formulation includes different types of carriers depending on whether the formulation is to be administered in solid, liquid, or aerosol form and whether it needs to be sterilized for an administration route such as injection.

[0240] In some embodiments, the modified Cav-1 peptide or its pharmaceutical formulation is administered intravenously, intrathecally, intradermally, transdermally, intrathecally, intraarterially, intraperitoneally, intranasally, intravaginally, rectally, intramuscularly, subcutaneously, mucosally, orally, topically, locally, by inhalation (e.g., inhalation of a spray or dry powder formulation), by injection, by infusion, by continuous infusion, by local perfusion directly immersing the target cells, via a catheter, via lavage, in a lipid composition (e.g., liposome), or by other methods known to those of skill in the art or any combination of the above (see, e.g., 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 of skill in the art based on the injection site, syringe passageability, and injectability, which includes consideration of the viscosity of the solution or suspension being injected as well as the drug concentration, pH, and weight osmolarity. In some cases, the particle size of the active agent can be selected to provide the desired dissolution rate upon administration (e.g., by subcutaneous injection).

[0241] In some embodiments, the formulations disclosed herein are administered intravenously, intramuscularly, or subcutaneously. In some embodiments, the formulations disclosed herein are administered by injection. In some embodiments, the formulations disclosed herein are sustained release, controlled release, or long - term release formulations.

[0242] In some embodiments, the formulation is an inhalable modified Cav - 1 peptide formulation. Administration via inhalation includes, but is not limited to, the use of an inhaler or nebulizer.

[0243] In some embodiments, the modified Cav - 1 peptide or its pharmaceutical formulation is formulated into the formulation in free base, neutral, or salt form. Pharmaceutically acceptable salts include acid addition salts, such as salts formed with the free amino groups of proteinaceous formulations, or salts formed with inorganic acids such as hydrochloric acid or phosphoric acid, or 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. When formulated, the solution is administered in a manner compatible with the dosage form and in an amount that is therapeutically effective. The formulation can be easily administered in various dosage forms, such as those formulated for parenteral administration like injection solutions, or aerosols for delivery to the lungs, or formulations for administration of nutrients such as drug - releasing capsules.

[0244] In some embodiments, the modified Cav-1 peptide or its pharmaceutical formulation comprises a pharmaceutically acceptable carrier, with or without an inert diluent. The carrier should be assimilable and includes liquids, semi-solids, i.e., pastes, or solid carriers. Its use in an administrable formulation for carrying out the method is appropriate, except when any conventional medium, agent, diluent, or carrier is harmful to the recipient or to the therapeutic effectiveness of the formulation contained therein. Examples of carriers or diluents include fats, oils, water, saline solutions, lipids, liposomes, resins, binders, fillers, etc., or combinations thereof. In some embodiments, the modified Cav-1 peptide or its pharmaceutical formulation includes one or more antioxidants to delay the oxidation of one or more components in the formulation. Additionally, 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, can provide protection against the action of microorganisms.

[0245] In some embodiments, the modified Cav-1 peptide or its pharmaceutical formulation is completely combined or mixed with a semi-solid or solid carrier. The mixing can be carried out by any convenient method such as grinding. Stabilizers can also be added to the mixing process to protect the formulation from loss of therapeutic activity, e.g., denaturation in the stomach. Examples of stabilizers include buffers, amino acids such as glycine and lysine, carbohydrates or cryoprotectants, such as dextrose, mannose, galactose, fructose, lactose, sucrose, maltose, sorbitol, mannitol, etc., but are not limited thereto.

[0246] In some embodiments, the modified Cav-1 peptide or its pharmaceutical formulation comprises one or more surfactants. Surfactants used according to the disclosed methods include ionic and non-ionic surfactants. Representative non-ionic surfactants include polysorbates, such as TWEEN®-20 and TWEEN-80® surfactants (ICI Americas Inc. of Bridgewater, N.J.); poloxamers (e.g., poloxamer 188); anionic and non-ionic surfactants, such as TRITON® surfactants (Sigma of St. Louis, Mo.); sodium dodecyl sulfate (SDS); sodium lauryl sulfate; sodium octyl glucoside; lauryl-, myristyl-, linoleyl-, or stearyl-sulphobetaine; lauryl-, myristyl-, linoleyl-, or stearyl-sarcosine; linoleyl-, myristyl-, or cetyl-betaine; lauramidopropyl-, cocamidopropyl-, linoleamidopropyl-, myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-betaine (e.g., lauramidopropyl); myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-dimethylamine; sodium methyl cocoyl-, or disodium methyl oleyl-taurate; cationic or quaternary phospholipid surfactants, such as MONAQUAT™ surfactants (Mona Industries Inc. of Paterson, N.J); polyethylene glycol, polypropylene glycol, block copolymers of ethylene glycol and propylene glycol, such as PLURONIC® surfactants (BASF of Mt. Olive, N.J.); oligo(ethylene oxide) alkyl ethers, alkyl (thio)glucosides, alkyl maltosides; and phospholipids.In some embodiments, one or more surfactants are present in the pharmaceutical formulation in an amount of about 0.01% to about 0.5% (weight of surfactant relative to the total weight of the 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 its pharmaceutical formulation is essentially free of nonionic surfactants or is essentially free of all surfactants.

[0247] With respect to the methods of treatment of the present invention, it is not intended that the administration of the modified Cav-1 peptide or its pharmaceutical formulation disclosed herein be limited to any particular mode of administration, dosage, or dosing frequency. The present invention contemplates all modes of administration, including intramuscular, intravenous, intraperitoneal, intracapsular, intra-articular, intralesional, subcutaneous, or any other route sufficient to provide an appropriate dosage for treating a disease or disorder. The modified Cav-1 peptide or its pharmaceutical formulation may be administered to a patient as a single dose or multiple doses. When multiple doses are administered, the doses may be separated from each other by, for example, 1 hour, 3 hours, 6 hours, 8 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 1 year, or any value or range therebetween. In some embodiments, the modified Cav-1 peptide or its pharmaceutical formulation is administered, for example, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 5 weeks, once every 6 weeks, once every 7 weeks, once every 8 weeks, once every 10 weeks, once every 15 weeks, once every 20 weeks, or once over a longer period. It should be understood that for any particular subject, the specific dosing regimen should be adjusted over time according to the individual needs and the professional judgment of the person administering or supervising the administration of the formulation. For example, the dosage of the modified Cav-1 peptide or its pharmaceutical formulation can be increased if sufficient therapeutic activity is not obtained at a lower dosage.

Examples

[0248] The present disclosure will be described in further detail by referring to the following examples. These examples are provided for illustrative purposes only and are not intended to be limiting unless otherwise specified. Thus, the present disclosure should in no way be construed as being limited to the following examples, but rather should be construed to encompass any and all variations that become apparent as a result of the teachings provided herein.

[0249] Example 1. SAIB-based Modified Cav-1 Peptide Formulation A formulation of SAIB-based modified Cav-1 peptide (APi2355, SEQ ID NO: 8) was prepared. Approximately 34.0 mg of micronized APi2355 was weighed and placed into a 1 mL syringe. Separately, approximately 567 mg of 80% w / w SAIB in NMP solution was weighed and placed into a separate 1 mL syringe. A Luer-to-Luer connection was placed between the two syringes and mixed until a uniform white suspension was formed (30 mg of APi2355 / 400 mg of SAIB). Similarly, suspensions of 10 mg of APi2355 / 500 mg of SAIB, 30 mg of APi2355 / 500 mg of SAIB, and 50 mg of APi2355 / 500 mg of SAIB were formed.

[0250] Approximately 150 mg of the 30 mg of APi2355 / 400 mg of SAIB suspension was added to approximately 35 mL of phosphate buffered saline (PBS) and placed on a shaking shaker at 150 rpm at 37°C. 1 mL of the sample was removed at various time points for testing and replaced with freshly warmed PBS. The sample was filtered and analyzed by HPLC for the % release of APi2355. The in vitro release kinetic parameters of the 30 mg of APi2355 / 5400 mg of SAIB formulation were determined as shown in Figure 1.

[0251] Example 2. Pharmacokinetics of Modified Cav-1 Peptide SAIB Formulation The pharmacokinetics of the Cav-1 peptide (APi2355) SAIB formulation prepared according to Example 1 (obtained with 7% w / w APi2355 vs 93% w / w SAIB with 5.66% w / w APi2355, 75.47% w / w SAIB and 18.87% w / w NMP) were evaluated over 42 days following a single-dose subcutaneous administration to male Sprague Dawley rats. One of three batches of the APi2355 SAIB formulation prepared according to Example 1 was administered to rats (n = 6) at a dose of 2.0 mg / kg APi2355 (concentration 60.0 mg / mL) in 0.033 mL / kg. The control group of rats (n = 2) received an injection of a vehicle formulation without APi2355.

[0252] Experimental procedures Animals were assigned to groups by a stratified randomization scheme designed to obtain similar mean body weights within groups. Animals were acclimated to the designated housing room for at least 5 days prior to dosing. All formulations were administered as a single dose by dorsal subcutaneous (SC) injection on Day 0. In the APi2355 treatment group, whole blood samples (40 - 50 μl) were collected from all animals before dosing and at 1 hour, 2 hours, 4 hours, 8 hours, 1 day, 2 days, 4 days, 7 days, 10 days, 14 days, 21 days, 29 days, and 35 days after treatment. In the control group, whole blood samples (40 - 50 μl, by tail snip) were collected from all animals before dosing and at 1 hour and 1 day after treatment. Blood was collected into K2EDTA tubes. After collection, the tubes were gently mixed and centrifuged at 3000 rpm and 4 °C for cooling within 30 minutes after collection. The resulting plasma was separated and transferred to uniquely labeled polypropylene tubes and frozen at -20 °C. Plasma and tissue samples were analyzed for the concentration of APi2355 using a qualified analytical procedure for LC / MS / MS.

[0253] Terminal whole blood samples (maximum volume by cardiac puncture) and tissues were collected from all animals on Day 42 after treatment. Tissues included the heart, liver, kidney, lung, and tissue from the skin compartment at the injection site. Additionally, the lungs were processed for bronchoalveolar lavage samples. Tissue samples were analyzed for APi2355 by LC / MS / MS.

[0254] Pharmacokinetic results In the control group, all concentration values of APi2355 at the pre-dose time point were below the lower limit of quantification (<0.3 ng / mL). As presented in Table 4, the concentration-time profile of APi2355 in rat plasma for the APi2355 treatment group was determined. As shown in Figure 2 and Table 5, the mean concentration profiles of APi2355 in various tissues were also determined. Table 6 summarizes the mean pharmacokinetic exposure profile of APi2355 in rat plasma measured after administration of the SAIB formulation.

[0255]

Table 4

[0256]

Table 5

[0257]

Table 6

[0258] The pharmacokinetic clearance and volume of distribution measured after administration of the SAIB formulation are summarized in Table 7.

[0259]

Table 7

[0260] Pharmacokinetic profile After single subcutaneous administration, APi2355 was absorbed from the SAIB formulation, and the median T max value was 0.083 days, and the mean C max value after injection was 477 ng / mL. APi2355 was detectable in plasma in 4 out of 6 rats after administration. The mean concentration at the last quantifiable time point was 0.942 ng / mL. As shown in Table 6, the mean AUC 0-14 value was 126 day*ng / mL, AUC0-22 Average value of 146 days * ng / mL, and AUC inf An average value of 183 days * ng / mL was observed.

[0261] AUC inf Since the extrapolated AUC was less than 20%, the time points used for the evaluation of APi2355 concentration appropriately explained the systemic exposure profile, and certain PK parameters such as AUC inf are considered to be accurate. The adjusted R-squared value was generally less than 0.8 due to the variability across the span, which was generally more than three times the half-life. The concentration of APi2355 varied significantly across the span, which led to the decrease in the adjusted R-squared. The half-life value showed an average of 9.24 days. The average MRT last value of the SAIB formulation was 7.82 days. Approximately 2 / 3 of the total exposure of APi2355 occurred within the first three weeks after SC administration.

[0262] Numbered embodiments Embodiment 1. A formulation comprising: a) a polypeptide comprising an amino acid sequence having a core sequence of FTTFTVT; and b) sucrose acetate isobutyrate (SAIB).

[0263] Embodiment 2. The formulation according to Embodiment 1, further comprising a first solvent.

[0264] Embodiment 3. The formulation according to Embodiment 2, wherein the first solvent is selected from N-methylpyrrolidone (NMP), absolute ethanol, or ethyl acetate.

[0265] Embodiment 4. The formulation according to any one of Embodiments 1 to 3, wherein the SAIB is present in the first solvent at about 50% w / w to about 95% w / w.

[0266] Embodiment 5. The formulation according to any one of Embodiments 1 to 4, wherein the SAIB is present in the first solvent at about 70% w / w to about 90% w / w.

[0267] Embodiment 6. The preparation according to any one of Embodiments 1 to 5, wherein the SAIB is present in the first solvent at about 80% w / w.

[0268] Embodiment 7. The preparation according to any one of Embodiments 1 to 6, wherein the first solvent is NMP or ethyl acetate.

[0269] Embodiment 8. The preparation according to any one of Embodiments 1 to 7, wherein the polypeptide is micronized.

[0270] Embodiment 9. The preparation according to any one of Embodiments 1 to 8, wherein the average particle size of the polypeptide ranges from about 0.5 μm to about 100 μm.

[0271] Embodiment 10. The preparation according to any one of Embodiments 1 to 9, wherein the average particle size of the polypeptide ranges from about 1 μm to about 5 μm.

[0272] Embodiment 11. The preparation according to any one of Embodiments 1 to 10, wherein the preparation is in the form of an emulsion, solution, microsphere, nanoparticle, nanosphere, implant, or gel.

[0273] Embodiment 12. The preparation according to any one of Embodiments 1 to 10, wherein the preparation is a suspension.

[0274] Embodiment 13. The preparation according to any one of Embodiments 1 to 12, further comprising a second solvent.

[0275] Embodiment 14. The preparation according to Embodiment 13, wherein the second solvent is sterile water, phosphate buffered saline (PBS), or any pharmaceutically acceptable carrier.

[0276] Embodiment 15. The preparation according to any one of Embodiments 1 to 14, wherein the preparation comprises the polypeptide and the SAIB in a weight ratio of about 1:50 to about 1:1.

[0277] Formulation according to Embodiment 15, wherein the weight ratio is from about 1:30 to about 1:1.

[0278] Formulation according to any one of Embodiments 1 to 16, wherein the formulation releases at least about 1% to about 30% of the polypeptide within 4 hours under physiological conditions.

[0279] Formulation according to any one of Embodiments 1 to 16, wherein the formulation releases at least about 5% to about 25% of the polypeptide within 4 hours under physiological conditions.

[0280] Formulation according to any one of Embodiments 1 to 18, wherein the polypeptide contains ASFTTFTVTK.

[0281] Formulation according to any one of Embodiments 1 to 19, wherein the polypeptide contains a) at least one amino acid added to the N-terminus, b) at least one amino acid added to the C-terminus, or c) at least one amino acid added to both the N-terminus and the C-terminus.

[0282] Formulation according to any one of Embodiments 1 to 20, wherein the polypeptide consists of 20 or fewer amino acids.

[0283] Formulation according to any one of Embodiments 1 to 21, wherein the addition made within 5 amino acids of each terminus has an amino acid sequence that is less than 80% identical to the contiguous amino acid sequence of SEQ ID NO: 1.

[0284] Formulation according to Embodiment 22, wherein the addition made within 5 amino acids of each terminus has an amino acid sequence that is less than 60% identical to the contiguous amino acid sequence of SEQ ID NO: 1.

[0285] Formulation according to Embodiment 22, wherein the addition made within 5 amino acids of each terminus has an amino acid sequence that is less than 40% identical to the contiguous amino acid sequence of SEQ ID NO: 1.

[0286] Embodiment 25. The formulation according to embodiment 22, wherein the addition made within 5 amino acids of each end has an amino acid sequence that is less than 20% identical to the contiguous amino acid sequence of SEQ ID NO: 1.

[0287] Embodiment 26. The formulation according to any one of embodiments 1 to 25, wherein the peptide sequence comprises a) the polypeptide comprises L-amino acids, b) the polypeptide comprises D-amino acids, or c) the polypeptide comprises both L-amino acids and D-amino acids.

[0288] Embodiment 27. The formulation according to any one of embodiments 1 to 26, wherein a) the polypeptide comprises at least one non-standard amino acid, or b) the polypeptide comprises two non-standard amino acids.

[0289] Embodiment 28. The formulation according to embodiment 27, wherein the non-standard amino acid is ornithine.

[0290] Embodiment 29. The formulation according to any one of embodiments 1 to 28, wherein the polypeptide further comprises a) an N-terminal modification, b) a C-terminal modification, or c) an N-terminal and C-terminal modification.

[0291] Embodiment 30. The formulation according to embodiment 29, wherein the N-terminal modification is acylation and / or the C-terminal modification is amidation.

[0292] Embodiment 31. The formulation according to any one of embodiments 1 to 30, wherein the polypeptide comprises the amino acid sequence of KASFTTFTVTKGS (SEQ ID NO: 4), KASFTTFTVTKGS-NH2 (SEQ ID NO: 5), aaEGKASFTTFTVTKGSaa (SEQ ID NO: 6), aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 7), Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8), OASFTTFTVTOS (SEQ ID NO: 9), or OASFTTFTVTOS-NH2 (SEQ ID NO: 10).

[0293] Embodiment 32. The preparation according to any one of Embodiments 1 to 31, wherein the polypeptide consists of an amino acid sequence selected from KASFTTFTVTKGS (SEQ ID NO: 4), KASFTTFTVTKGS-NH2 (SEQ ID NO: 5), aaEGKASFTTFTVTKGSaa (SEQ ID NO: 6), aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 7), Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8), OASFTTFTVTOS (SEQ ID NO: 9), or OASFTTFTVTOS-NH2 (SEQ ID NO: 10).

[0294] Embodiment 33. The preparation according to any one of Embodiments 1 to 32, wherein the polypeptide consists of the amino acid sequence of Ac-aaEGKASFTTFTVTKGSaa-NH2 (SEQ ID NO: 8).

[0295] Embodiment 34. A method of performing it on a subject in need of treatment of a disease, the method comprising administering to the subject a pharmaceutically effective amount of the preparation according to any one of Embodiments 1 to 33.

[0296] Embodiment 35. The method according to Embodiment 34, wherein the disease is fibrosis.

[0297] Embodiment 36. The fibrosis is interstitial lung disease, liver fibrosis, kidney fibrosis, skin fibrosis, glomerulonephritis, systemic sclerosis, cardiac fibrosis, myocardial fibrosis, renal fibrosis, liver cirrhosis, nephrosclerosis, arteriosclerosis, macular degeneration, ocular scarring, cataract, retinal and vitreoretinopathy, Graves ophthalmopathy, neurofibromatosis, scleroderma, glioblastoma, keloid and hypertrophic scarring, peritoneal fibrotic disease, chronic obstructive pulmonary disease, postoperative myoma, diabetic nephropathy, gynecological cancer, myeloproliferative syndrome, myeloid leukemia, myelodysplastic syndrome, inflammatory bowel disease, non-alcoholic fatty liver disease, fibrosarcoma, rheumatoid arthritis, non-alcoholic steatohepatitis, Alport syndrome, or chronic COVID syndrome. The method according to Embodiment 35.

[0298] Method according to embodiment 36, wherein the interstitial lung disease is idiopathic pulmonary fibrosis, familial pulmonary fibrosis, idiopathic nonspecific interstitial pneumonia, conventional interstitial pneumonia, idiopathic organizing pneumonia, or sarcoidosis.

[0299] Method according to embodiment 37, wherein the interstitial lung disease is idiopathic pulmonary fibrosis.

[0300] Method according to embodiment 34, wherein the disease is a renal disease or a renal disorder.

[0301] Method according to embodiment 39, wherein the renal disease or the renal 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.

[0302] Method according to embodiment 40, wherein the renal disease or the renal disorder is renal fibrosis.

[0303] Method according to embodiment 40, wherein the renal disease or the renal disorder is Alport syndrome.

[0304] Method according to any one of embodiments 34 to 42, wherein the administration is intravitreal, intradermal, transdermal, intramuscular, or subcutaneous administration.

[0305] Method according to any one of embodiments 34 to 43, wherein the subject is a human.

[0306] Method according to any one of embodiments 34 to 44, wherein the formulation releases the polypeptide for at least 7 days after a single-dose administration.

[0307] Embodiment 46. The method according to any one of Embodiments 34 to 44, wherein the formulation releases the polypeptide for at least 21 days after administration of a single dose.

[0308] Embodiment 47. The method according to any one of Embodiments 34 to 44, wherein the formulation releases the polypeptide for at least 28 days after administration of a single dose.

[0309] Embodiment 48. The method according to any one of Embodiments 34 to 47, wherein the formulation is administered to the subject at a dose of about 0.01 mg / kg to about 250 mg / kg.

[0310] Embodiment 49. The method according to Embodiment 48, wherein the formulation is administered to the subject at a dose of about 0.05 mg / kg to about 50 mg / kg.

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