Ionic self-assembling peptides
Provided herein are ionic self-assembling peptides, pharmaceutical compositions comprising the peptides, and methods of using and making the same.
1 . A pharmaceutical composition comprising a self-assembling peptide in aqueous solution, wherein the self-assembling peptide self-assembles when exposed to physiological conditions thereby forming a hydrogel, wherein the self-assembling peptide comprises the amino acid sequence selected from the group consisting of:
(a) SEQ ID NO:1 and SEQ ID NO:8, the self-assembling peptide having a net charge of +3 when formulated in a pharmaceutical composition at a pH of 7.4; and
(b) SEQ ID NO:16 and SEQ ID NO:94, the self-assembling peptide having a net charge of −3 when formulated in a pharmaceutical composition at a pH of 7.4.
2 . The pharmaceutical composition of claim 1 , wherein the self-assembling peptide comprises one or both of:
a) an N-terminal functional group selected from the group consisting of an acetyl, a formyl, pyroglutamyl (pGlu), biotin, polyethylene glycol (PEG), urea, alkylamine, a carbamate, a sulfonamide, dansyl, 2,4-dintrophenyl, fluorescein, 7-methoxycoumarin acetic acid, 9-fluorenylmethyloxycarbonyl, succinyl, chloroacetyl, maleimide, benzyloxycarbonyl, bromoacetyl, nitrilotriacetyl, tertbutoxycarbonyl, 4-hydroxyphenylpropionic acid, allyloxycarbonyl, a fatty acid, and trityl, and
b) a C-terminal functional group selected from the group consisting of an amine, an amido, an N-alkyl amide, an aldehyde, an ester, an alcohol, para-nitroanilide (pNA), 7-amino-4-methylcoumarin (Amc), a hydrazide, hydroxamic acid, chloromethylketone, p-nitroaniline, para-nitrophenol, fluoromethylketone, and cysteamide.
3 . The pharmaceutical composition of claim 1 , wherein the self-assembling peptide further comprises at least one biologically active peptide motif at the N-terminal end, or the C-terminal end, or both, of the self-assembling peptide.
4 . The pharmaceutical composition of claim 3 , wherein the at least one biologically active peptide motif is derived from laminin-1, collagen IV, fibronectin, elastin, bone marrow homing peptide 1, bone marrow homing peptide 2, or myelopeptide.
5 . The pharmaceutical composition of claim 1 , further comprising a tonicity agent wherein the tonicity agent comprises:
one or more salts selected from the group consisting of NaCl, KCl, MgCl 2 , CaCl 2 , NH 4 Cl, Na 2 HPO 4 , KH 2 PO 4 , and CaSO 4 ; and/or
one or more sugars selected from the group consisting of dextrose, mannitol, glycerin, sucrose, and trehalose.
6 . The pharmaceutical composition of claim 5 , wherein the tonicity agent comprises one or more salts and is present in a concentration of 0.01 M to 0.3 M.
7 . The pharmaceutical composition of claim 5 , wherein the tonicity agent comprises one or more sugars and is present in a concentration of 0.1-10% (w/v).
8 . The pharmaceutical composition of claim 1 , having a pH within the range from 6 to 8.
9 . The pharmaceutical composition of claim 1 , wherein the concentration of the self-assembling peptide is from 0.01% (w/v) to 10% (w/v).
10 . The pharmaceutical composition of claim 1 , further comprising an isolated cell.
11 . The pharmaceutical composition of claim 10 , wherein the isolated cell is a mammalian cell selected from the group consisting of: an immune cell, a stem cell, a chondrocyte progenitor cell, a pancreatic progenitor cell, a fibroblast, a keratinocyte, a neuronal cell, glial cell, pre-adipocyte, adipocyte, vascular endothelial cell, endothelial progenitor cell, mesenchymal cell, smooth muscle progenitor cell, cardiac myocyte, myoblast, and a capillary endothelial cell.
12 . The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition further comprises a bioactive agent.
13 . The pharmaceutical composition of claim 12 , wherein the bioactive agent is selected from the group consisting of a hormone, an enzyme, an siRNA, an shRNA, an antisense-RNA, an antibiotic, an antibody, and an anti-inflammatory agent.
14 . The pharmaceutical composition of claim 1 , wherein the hydrogel comprises a storage modulus of at least 10 Pascal (Pa).
15 . An article of manufacture comprising the pharmaceutical composition of claim 1 , wherein the article is a syringe, a vial, an auto-injector, tubing, or a catheter.
16 . A method of promoting tissue repair or regeneration in a subject in need thereof, comprising administering to a tissue of the subject an effective amount of the pharmaceutical composition according to claim 1 , thereby promoting tissue repair or regeneration of the tissue.
17 . The method of claim 16 , wherein the tissue is skin, bone, cartilage, neural, ligament, tendon, vascular tissue, optic, muscle, or cardiac tissue.
18 . The method of claim 16 , wherein the subject has a congenital disease or disorder resulting in a need for the tissue repair or regeneration; or the subject has suffered an injury resulting in a need for the tissue repair or regeneration, wherein the injury is a result of surgery, trauma, stroke, tumor, or a disease or disorder.
19 . A method of promoting wound healing in a subject in need thereof, comprising administering to a subject's wound an effective amount of the pharmaceutical composition according to claim 1 , thereby promoting wound healing and/or anti-microbial activity, wherein the wound comprises an abrasion, a burn, a chap, a detrition, a cut, an ulcer, a laceration, an incision, or a scratch.
20 . A method of reducing bleeding at a site within a subject, comprising administering to the site an effective amount of the pharmaceutical composition according to claim 1 , wherein the pharmaceutical composition creates a physical barrier thereby reducing bleeding at the site within the subject.
21 . A method of excising a lesion from a site in the gastrointestinal tract of a subject, comprising:
administering to the submucosa below the lesion the pharmaceutical composition according to claim 1 in an amount sufficient to lift the lesion; and
excising the lesion from the site in the gastrointestinal tract of the subject.
22 . The method of claim 21 , wherein the lesion comprises a polyp, an ulcer, or a tumor.
23 . The method of claim 21 , wherein the lesion is present in a region of the gastrointestinal tract selected from the group consisting of: mouth, throat, esophagus, stomach, small intestine, large intestine, colon, and rectum.
24 . A method of culturing a cell, comprising contacting the cell with the pharmaceutical composition according to claim 1 .
25 . A method of treating a pulmonary bulla in a subject, comprising:
introducing a delivery device to a target area of the pulmonary bulla of the subject;
positioning an end of the delivery device in the target area in which a treatment of the pulmonary bulla is desired;
administering, through the delivery device, the pharmaceutical composition according to claim 1 in an effective amount and in an effective concentration to the target area to form a barrier under physiological conditions of the target area to treat the pulmonary bulla;
removing the delivery device from the target area; and
collapsing the pulmonary bulla prior or subsequent to administering the solution.
26 . A method for mitigating adhesion to a biological tissue, the method comprising administering to the biological tissue an effective amount of the pharmaceutical composition according to claim 1 , to thereby mitigate adhesion to the biological tissue.
27 . A method of filling a bone void in a subject, comprising introducing a delivery device to a bone of a subject;
positioning an end of the delivery device proximate a void in the bone where promotion of bone growth is desired;
administering the pharmaceutical composition according to claim 1 through the delivery device; and
removing the delivery device, wherein the delivery device is chosen from a syringe, an auto-injector, a tubing, or a catheter.
28 . A method of treating dry eye in a subject, comprising administering to an eye of the subject an effective amount of the pharmaceutical composition according to claim 1 .
29 . A self-assembling peptide comprising the amino acid sequence set forth in SEQ ID NO:1.
30 . A self-assembling peptide comprising the amino acid sequence set forth in SEQ ID NO:16.
31 . A self-assembling peptide comprising the amino acid sequence set forth in SEQ ID NO:8.
32 . A self-assembling peptide comprising the amino acid sequence set forth in SEQ ID NO:94.
33 . The composition claim 1 , wherein the self-assembling peptide comprises the amino acid sequence set forth in SEQ ID NO:1.
34 . The composition of claim 1 , wherein the self-assembling peptide comprises the amino acid sequence set forth in SEQ ID NO:16.
35 . The composition of claim 1 , wherein the self-assembling peptide comprises the amino acid sequence set forth in SEQ ID NO:8.
36 . The composition of claim 1 , wherein the self-assembling peptide comprises the amino acid sequence set forth in SEQ ID NO:94.
37 . The method of claim 16 , wherein the self-assembling peptide comprises the amino acid sequence set forth in SEQ ID NO:1.
38 . The method of claim 16 , wherein the self-assembling peptide comprises the amino acid sequence set forth in SEQ ID NO: 16.
39 . The method of claim 16 , wherein the self-assembling peptide comprises the amino acid sequence set forth in SEQ ID NO:8.
40 . The method of claim 16 , wherein the self-assembling peptide comprises the amino acid sequence set forth in SEQ ID NO:94.
41 . The method of claim 11 , wherein the neuronal cell is an astrocyte.
42 . The method of claim 11 , wherein the keratinocyte is an epidermal keratinocyte.