STABLE PEPTIDES AND METHODS OF USE THEREOF

1 - 372 . (canceled)

373 . A method comprising administering a compound comprising a peptide to a subject, wherein at least 70% of the peptide remains intact after exposure to pepsin at a temperature of at least 20° C. for at least 5 minutes and the peptide comprises at least 6 cysteine residues.

374 . The method of claim 373 , wherein the peptide remains intact when exposed to pepsin at a concentration of 47 U/ml and a temperature of 23° C. for 5 minutes as measured by high performance liquid chromatography.

375 . The method of claim 373 , wherein the peptide has at least one of the following characteristics:

(a) at least 70% of the peptide remains intact after exposure to pepsin at a temperature of at least 23° C. for at least 5 minutes;

(b) at least 70% of the peptide remains intact after exposure to dithiothreitol (DTT) at a concentration of 5 mM and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(c) at least 70% of the peptide remains intact after exposure to reduced glutathione (GSH) at a concentration of 5 mM and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(d) at least 70% of the peptide remains intact after exposure to trypsin at a concentration of 0.5 U/ml and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(e) at least 70% of the peptide remains intact after exposure to simulated gastric fluid (SGF; pH 1.05; 2% (w/v) sodium chloride in 0.7% (v/v) hydrochloric acid) and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(f) at least 70% of the peptide remains intact after exposure to a pH of 5 and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(g) at least 70% of the peptide remains intact after passage through the mouth, stomach, small intestine, or the large intestine, as measured by tandem high performance liquid chromatography and liquid scintillation counting;

(h) at least 70% of the peptide remains intact after exposure to the combination of simulated gastric fluid (SGF; pH 1.05; 2% (w/v) sodium chloride in 0.7% (v/v) hydrochloric acid) with 0.5 U/ml pepsin, 100 mM Tris, and 10 mM DTT and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(i) at least 70% of the peptide remains intact after exposure to at least 70° C. for at least 5 minutes, as measured by high performance liquid chromatography; or

(j) at least 70% of the peptide remains intact after exposure to at least 100° C. for at least 5 minutes, as measured by high performance liquid chromatography.

376 . The method of claim 373 , wherein the peptide comprises three or more disulfide bridges formed between cysteine residues, wherein one of the disulfide bridges passes through a loop formed by two other disulfide bridges.

377 . The method of claim 373 , wherein the peptide comprises a structural proline amino acid residue.

378 . The method of claim 373 , wherein the peptide comprises at least 3 positively charged amino acid residues.

379 . The method of claim 373 , wherein the peptide comprises a sequence motif of leucine-X 1 -X 2 -leucine-phenylalanine (LX 1 X 2 LF).

380 . The method of claim 373 , wherein the peptide comprises:

a) at least 90% sequence identity to SEQ ID NO: 114; or

b) at least 90% sequence identity to any one of SEQ ID NO: 110, SEQ ID NO: 140, SEQ ID NO: 89, SEQ ID NO: 85; SEQ ID: 107, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 94, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 100, SEQ ID NO: 92, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 87, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 85, SEQ ID NO: 101, SEQ ID NO: 115, SEQ ID NO: 108, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 112, SEQ ID NO: 134, SEQ ID NO: 135, SEQ ID NO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, SEQ ID NO: 106, or SEQ ID NO: 88.

381 . The method of claim 373 , wherein the peptide achieves an average t½ of 0.1 hours-168 hours in a subject after administering the peptide to the subject.

382 . The method of claim 373 , wherein the compound is administered via oral administration, inhalation, intranasal administration, topical administration, intravenous administration, subcutaneous administration, intra-articular administration, intramuscular administration, intraperitoneal administration, intra-synovial administration, vaginal administration, rectal administration, pulmonary administration, ocular administration, buccal administration, sublingual administration, intrathecal administration, or any combination thereof, to a subject.

383 . The method of claim 373 , wherein the peptide is linked to an active agent.

384 . The method of claim 383 , wherein the active agent is: a peptide, an oligopeptide, a polypeptide, a polynucleotide, a polyribonucleotide, a DNA, a cDNA, a ssDNA, a RNA, a dsRNA, a micro RNA, an oligonucleotide, an antibody, an antibody fragment, an aptamer, a cytokine, an enzyme, a growth factor, a chemokine, a neurotransmitter, a chemical agent, a fluorophore, a metal, a metal chelate, an X-ray contrast agent, a PET agent, a radioisotope, a photosensitizer, a radiosensitizer, a radionuclide chelator, a therapeutic small molecule, a steroid, a corticosteroid, an anti-inflammatory agent, an immune modulator, a protease inhibitor, an amino sugar, a chemotherapeutic agent, a cytotoxic chemical, a toxin, a tyrosine kinase inhibitor, an anti-infective agent, an antibiotic, an anti-viral agent, an anti-fungal agent, an aminoglycoside, a nonsteroidal anti-inflammatory drug (NSAID), a statin, a nanoparticle, a liposome, a polymer, a biopolymer, a polysaccharide, a proteoglycan, a glycosaminoglycan, a glucocorticoid, an anti-cytokine agent, a pain-reducing agent, a dendrimer, a fatty acid, an Fc region, siderocalin, or any combination thereof.

385 . The method of claim 384 , wherein the steroid is triamcinolone, triamcinolone hexacetonide, budesonide, or dexamethasone.

386 . The method of claim 373 , wherein administering the compound treats the subject.

387 . The method of claim 373 , wherein the subject has a gastrointestinal infection or chronic gastrointestinal disease.

388 . The method of claim 387 , wherein the gastrointestinal infection is a bacterial infection, prokaryotic infection, yeast infection, or fungal infection.

389 . The method of claim 387 , wherein the chronic gastrointestinal disease is irritable bowel syndrome, inflammatory bowel disease, Crohn's disease, gastroesophageal reflux disease, ulcerative colitis, or constipation.

390 . The method of claim 373 , wherein the subject has a cancer, and wherein the cancer is colorectal cancer, stomach cancer, or esophageal cancer.

391 . The method of claim 373 , wherein the subject has an inflammation, a cancer, a degradation, a growth disturbance, genetic, a tear, an infection, an injury, a rheumatic condition, an immune system disorder, a kidney disease, lung disease, a condition of aging, a degenerative brain condition, a degenerative body condition, a childhood condition, a hepatic disease, a pulmonary disease, a pancreatic condition, or a gastrointestinal condition.

392 . The method of claim 391 , wherein the kidney disease is acute kidney injury or chronic kidney disease.

393 . The method of claim 373 , wherein the peptide homes to cartilage, kidneys, proximal tubules of the kidneys, or tumors.

394 . The method of claim 373 , wherein the peptide enters a cell, and wherein the peptide is active intracellularly.

395 . The method of claim 373 , wherein the peptide is formulated in a pharmaceutical composition.

396 . The method of claim 395 , wherein the pharmaceutical composition further includes a permeation enhancer and the permeation enhancer increases oral absorption.

397 . The method of claim 396 , wherein the permeation enhancer is SNAC, 5-CNAC, sodium caprylate, an aromatic alcohol, EDTA, a sodium alkyl sulfate, or a citrate.

398 . The method of claim 383 , wherein the peptide linked to the active agent brings or enters a cell and wherein the active agent is delivered to the cell or is active intracellularly.

399 . The method of claim 383 , wherein the peptide linked to the active agent is formulated in a pharmaceutical formulation.

400 . The method of claim 399 , wherein the pharmaceutical composition further includes a permeation enhancer and the permeation enhancer increases oral absorption.

401 . The method of claim 400 , wherein the permeation enhancer is SNAC, 5-CNAC, sodium caprylate, an aromatic alcohol, EDTA, a sodium alkyl sulfate, or a citrate.

402 . A compound comprising a peptide for use in a treatment of a gastrointestinal disorder, wherein at least 70% of the peptide remains intact after exposure to pepsin at a temperature of at least 20° C. for at least 5 minutes and the peptide comprises at least 6 cysteine residues.

403 . The peptide of claim 402 having at least one of the following characteristics:

(a) at least 70% of the peptide remains intact after exposure to pepsin at a temperature of at least 23° C. for at least 5 minutes;

(b) at least 70% of the peptide remains intact after exposure to dithiothreitol (DTT) at a concentration of 5 mM and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(c) at least 70% of the peptide remains intact after exposure to reduced glutathione (GSH) at a concentration of 5 mM and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(d) at least 70% of the peptide remains intact after exposure to trypsin at a concentration of 0.5 U/ml and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(e) at least 70% of the peptide remains intact after exposure to simulated gastric fluid (SGF; pH 1.05; 2% (w/v) sodium chloride in 0.7% (v/v) hydrochloric acid) and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(f) at least 70% of the peptide remains intact after exposure to a pH of 5 and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(g) at least 70% of the peptide remains intact after passage through the mouth, stomach, small intestine, or the large intestine, as measured by tandem high performance liquid chromatography and liquid scintillation counting;

(h) at least 70% of the peptide remains intact after exposure to the combination of simulated gastric fluid (SGF; pH 1.05; 2% (w/v) sodium chloride in 0.7% (v/v) hydrochloric acid) with 0.5 U/ml pepsin, 100 mM Tris, and 10 mM DTT and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(i) at least 70% of the peptide remains intact after exposure to at least 70° C. for at least 5 minutes, as measured by high performance liquid chromatography; or

(j) at least 70% of the peptide remains intact after exposure to at least 100° C. for at least 5 minutes, as measured by high performance liquid chromatography.

404 . A compound comprising a peptide and a permeation enhancer, wherein at least 70% of the peptide remains intact after exposure to pepsin at a temperature of at least 20° C. for at least 5 minutes and the peptide comprises at least 6 cysteine residues.

405 . The peptide of claim 404 having at least one of the following characteristics:

(a) at least 70% of the peptide remains intact after exposure to pepsin at a temperature of at least 23° C. for at least 5 minutes;

(b) at least 70% of the peptide remains intact after exposure to dithiothreitol (DTT) at a concentration of 5 mM and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(c) at least 70% of the peptide remains intact after exposure to reduced glutathione (GSH) at a concentration of 5 mM and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(d) at least 70% of the peptide remains intact after exposure to trypsin at a concentration of 0.5 U/ml and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(e) at least 70% of the peptide remains intact after exposure to simulated gastric fluid (SGF; pH 1.05; 2% (w/v) sodium chloride in 0.7% (v/v) hydrochloric acid) and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(f) at least 70% of the peptide remains intact after exposure to a pH of 5 and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(g) at least 70% of the peptide remains intact after passage through the mouth, stomach, small intestine, or the large intestine, as measured by tandem high performance liquid chromatography and liquid scintillation counting;

(h) at least 70% of the peptide remains intact after exposure to the combination of simulated gastric fluid (SGF; pH 1.05; 2% (w/v) sodium chloride in 0.7% (v/v) hydrochloric acid) with 0.5 U/ml pepsin, 100 mM Tris, and 10 mM DTT and a temperature of at least 20° C. for at least 5 minutes, as measured by high performance liquid chromatography;

(i) at least 70% of the peptide remains intact after exposure to at least 70° C. for at least 5 minutes, as measured by high performance liquid chromatography; or

(j) at least 70% of the peptide remains intact after exposure to at least 100° C. for at least 5 minutes, as measured by high performance liquid chromatography.

406 . The peptide of claim 404 , wherein the permeation enhancer is SNAC, 5-CNAC, sodium caprylate, an aromatic alcohol, EDTA, a sodium alkyl sulfate, or a citrate.