Ionic self-assembling peptides

Abstract

To provide ionic self-assembling peptides.SOLUTION: Provided herein are ionic self-assembling peptides, pharmaceutical compositions comprising the peptides, and methods of using and making the same. Pharmaceutical compositions comprising the ionic self-assembling peptides described herein may be formulated at neutral pH and administered to a subject without inducing tissue damage that may be associated with pharmaceutical compositions having acidic or basic pH. Moreover, the ionic self-assembling peptides can be used to make improved hydrogels having advantageous physical characteristics under physiological conditions.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Claim of priority This application claims the benefits of U.S. Provisional Application No. 62 / 693877, filed on 3 July 2018. The entirety of the foregoing is incorporated herein by reference.

[0002] Sequence List This application includes a sequence listing, which is filed electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy, created on July 1, 2019, is named 46406-0029WO1 SEQ.txt and has a size of 24576 bytes.

[0003] Field of Invention This disclosure generally relates to self-assembling peptides, pharmaceutical compositions containing self-assembling peptides, and methods for using them. [Background technology]

[0004] background Self-assembling peptides have been developed for a variety of purposes, including scaffolds for tissue engineering and regenerative medicine, drug delivery, three-dimensional tissue culture, and hemostasis. Examples of such self-assembling peptides include (1) alternating positive and negative charges and hydrophobic residues (e.g., RADA16 (SEQ ID NO: 91), IEIK13 (SEQ ID NO: 92), and KLDL12 (SEQ ID NO: 93)), (2) alternating sequences of nonionic polar residues and hydrophobic residues, and (3) β-sheet peptides having repeating nonionic polar residues. However, such self-assembling peptides have practical limitations. For example, pharmaceutical compositions containing these self-assembling peptides with alternating positive and negative charges and hydrophobic residues (e.g., RADA16 (SEQ ID NO: 91)) must be formulated at an acidic pH to solubilize the peptide, which may cause cell and / or tissue damage upon administration to the subject. Therefore, there is still a need for improved self-assembling peptides that can be used for therapeutic purposes. [Overview of the project] [Means for solving the problem]

[0005] Summary of the Invention This disclosure is at least in part based on the development of ionic self-assembling peptides having specific combinations of ionic polar amino acids, hydrophobic amino acids, and nonionic polar amino acids. Previously described self-assembling peptides typically contain equal amounts of amino acid residues having a net negative charge and a net positive charge at neutral pH. These peptides are formulated at acidic or basic pH to charge the peptides so that they remain in solution and compositions containing the peptides are fluid and injectable. When formulated at neutral pH, phase separation and precipitation of the peptides have been observed. For example, RADA16 (SEQ ID NO: 91) has four positive charges and four negative charges at neutral pH. When formulated at pH 7.5, as shown in Figure 20A, RADA16 (SEQ ID NO: 91) precipitates.

[0006] In contrast, including ionic polar amino acids in the self-assembling peptides provided herein yields peptides with a non-zero net charge at neutral pH, thereby enabling the formulation of the peptides at neutral pH. Advantageously, the ionic self-assembling peptides provided herein can be formulated at neutral pH and remain soluble and stable in solution. While not wishing to be bound by any particular theory, pharmaceutical compositions containing the ionic self-assembling peptides described herein can be formulated at neutral pH and administered to subjects without inducing tissue damage that may be associated with pharmaceutical compositions having acidic or basic pH. Furthermore, ionic self-assembling peptides can be used to prepare improved hydrogels with advantageous physical properties under physiological conditions. For example, aqueous solutions of self-assembling peptides that remain stable under physiological conditions, are fluid, and injectable can be produced. Upon gelation, the self-assembling peptides can form hydrogels with shear-thickening, thixotropic, and rheological properties useful for clinical, industrial, and / or research applications.

[0007] In some embodiments, the Disclosure provides self-assembling peptides and pharmaceutical compositions comprising the following amino acid sequences: [(X)i(Y)j(Z)k(Y)l]m(X)n (Formula I), [(Y)i(X)j(Y)k(Z)l]m(Y)n (Formula II), [(Z)i(Y)j(X)k(Y)l]m(Z)n (Formula III), or [(Y)i(Z)j(Y)k(X)l]m(Y)n (Formula IV) (wherein each (X) is independently an ionic polar amino acid, each (Y) is independently a hydrophobic amino acid, each (Z) is independently a nonionic polar amino acid, each i, j, k and l is independently an integer ≥ 1, m is an integer ≥ 2, and n = 0 or an integer ≥ 1). In some embodiments, at least one of i, j, k and l is independently an integer 1. In some embodiments, at least one of i, j, k, and l is independently an integer 2. In some embodiments, each of i, j, k, and l is 1. In some embodiments, m is independently an integer 2, 3, or 4.

[0008] In some embodiments, the self-assembling peptide includes or consists of the amino acid sequence shown in formula I. In some embodiments, the self-assembling peptide includes or consists of the amino acid sequence shown in formula II. In some embodiments, the self-assembling peptide includes or consists of the amino acid sequence shown in formula III. In some embodiments, the self-assembling peptide includes or consists of the amino acid sequence shown in formula IV.

[0009] In some embodiments, each (X) is a basic amino acid (e.g., arginine, lysine, histidine, or ornithine). In some embodiments, each (X) is an acidic amino acid (e.g., aspartic acid or glutamic acid).

[0010] In some embodiments, each (Y) is one of alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, or glycine.

[0011] In some embodiments, each (Z) is one of serine, threonine, tyrosine, cysteine, glutamine, asparagine, or methionine.

[0012] In some embodiments, the self-assembling peptide comprises or consists of the amino acid sequences shown in SEQ ID NOs: 1-20 and 94-96.

[0013] In some embodiments, the self-assembling peptide includes an N-terminal functional group, a C-terminal functional group, or both. In some embodiments, the N-terminal functional group is any of acetyl, formyl, pyroglutamyl (pGlu), biotin, polyethylene glycol (PEG), urea, alkylamine, carbamate, sulfonamide, dansyl, 2,4-dinitrophenyl (2,4-dintrophenyl), fluorescein, 7-methoxycoumarin acetate, 9-fluorenylmethyloxycarbonyl, palmitic acid, succinyl, chloroacetyl, maleimide, benzyloxycarbonyl, bromoacetyl, nitrilotriacetyl, tert-butoxycarbonyl, 4-hydroxyphenylpropionic acid, allyloxycarbonyl, butyric acid, fatty acid, or trityl. In some embodiments, the C-terminal functional group is one of the following: amide, N-alkylamide, aldehyde, ester, alcohol, para-nitroanilide (pNA), 7-amino-4-methylcoumarin (Amc), hydrazide, hydroxamic acid, chloromethyl ketone, p-nitroaniline, para-nitrophenol, hydroxysucinimide ester, fluoromethyl ketone, cysteamide, 9-fluorenemethyl (Fm) ester, allyl ester, 2,4-dimethoxybenzyl ester, 2-phenylisopropyl ester, p-nitrobenzyl ester, and 2-chlorotrityl ester. In some embodiments, the self-assembling peptide comprises or consists of the amino acid sequences shown in SEQ ID NOs. 21-40 and 97-99.

[0014] In some embodiments, the self-assembling peptide comprises at least one (e.g., one, two, three, four, or more) biologically active peptide motifs. In some embodiments, at least one biologically active peptide motif is located at the N-terminus of the self-assembling peptide. In some embodiments, at least one biologically active peptide motif is located at the C-terminus of the self-assembling peptide. In some embodiments, 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. In some embodiments, at least one biologically active peptide motif comprises or consists of the amino acid sequences shown in any one of SEQ ID NOs: 41-70. In some embodiments, the self-assembling peptide comprises or consists of the amino acid sequences shown in SEQ ID NOs: 71-90.

[0015] In some embodiments, the pharmaceutical composition contains an isotonic agent. In some embodiments, the isotonic agent contains one or more salts selected from the group consisting of NaCl, KCl, MgCl2, CaCl2, NH4Cl, Na2HPO4, KH2PO4, and CaSO4. In some embodiments, the isotonic agent further contains one or more sugars selected from the group consisting of dextrose, mannitol, glycerin, sucrose, and trehalose. In some embodiments (for example, when the isotonic agent is one or more salts), the isotonic agent is present at a concentration of about 0.01 M to about 0.3 M. In some embodiments (for example, when the isotonic agent is one or more salts), the isotonic agent is present at a concentration of about 0.15 M. In some embodiments (for example, when the isotonic agent is one or more sugars), the isotonic agent is present at a concentration of about 0.1 to 10% (w / v). In some embodiments (for example, when the isotonic agent is a sugar of 1 or more), the isotonic agent is present at a concentration of about 10% (w / v). In some embodiments, the isotonic agent enhances the rheological properties of the composition or hydrogel peptide comprising the self-assembling peptide described herein.

[0016] In some embodiments, the pharmaceutical composition has a pH of about 6 to about 8. In some embodiments, the pharmaceutical composition has a pH of about 7 to about 7.5. In some embodiments, the net charge of the self-assembling peptide in the pharmaceutical composition is greater than or equal to +1 or less than or equal to -1. In some embodiments, the net charge of the self-assembling peptide in the pharmaceutical composition is about +1 to about +6 (e.g., +1, +2, +3, +4, +5, or +6). In some embodiments, the net charge of the self-assembling peptide in the pharmaceutical composition is about -1 to about -6 (e.g., -1, -2, -3, -4, -5, or -6).

[0017] In some embodiments, the concentration of the self-assembling peptide in the pharmaceutical composition is about 0.01% (w / v) to about 10% (w / v). In some embodiments, the concentration of the self-assembling peptide in the pharmaceutical composition is about 0.1% (w / v) to about 5% (w / v). In some embodiments, the concentration of the self-assembling peptide in the pharmaceutical composition is about 0.5% (w / v) to about 1.5% (w / v). In some embodiments, the concentration of the self-assembling peptide in the pharmaceutical composition is about 1% (w / v).

[0018] In some embodiments, the pharmaceutical composition comprises isolated cells (e.g., stem cells). In some embodiments, the isolated cells are mammalian cells. In some embodiments, the mammalian cells are immune cells, stem cells, chondrocyte progenitor cells, pancreatic progenitor cells, myoblasts, fibroblasts, keratinocytes, nerve cells, glial cells, astrocytes, preadipocytes, adipocytes, vascular endothelial cells, endothelial progenitor cells, mesenchymal cells, neural stem cells, immune cells (e.g., B cells and T cells), smooth muscle progenitor cells, cardiomyocytes, fetal dermal fibroblasts, epidermal keratinocytes, myoblasts, and capillary endothelial cells.

[0019] In some embodiments, the pharmaceutical composition comprises at least one (e.g., one, two, three, four, five or more) bioactive agent. In some embodiments, the bioactive agent is a hormone, growth factor, insulin, enzyme, siRNA, shRNA, antisense RNA, antisense DNA, mRNA, antibiotic, antibody or anti-inflammatory agent.

[0020] In some embodiments, the pharmaceutical composition is an aqueous solution. In some embodiments, the pharmaceutical composition is a hydrogel.

[0021] In some embodiments, the pharmaceutical composition is a hydrogel having a storage modulus of at least about 10 Pascals (Pa) (e.g., about 25 Pa, about 50 Pa, about 100 Pa, about 150 Pa, about 250 Pa, about 500 Pa, about 750 Pa, about 1000 Pa or more).

[0022] In another aspect, the present disclosure provides an article of manufacture comprising the self-assembling peptides or pharmaceutical compositions described herein. In some embodiments, the article is a syringe, vial, auto-injector device, tube or catheter.

[0023] In another aspect, the present disclosure provides a method of treating a subject in need of treatment, the method comprising administering to the subject an effective amount of the self-assembling peptides or pharmaceutical compositions described herein.

[0024] In yet another embodiment, the Disclosure provides a method for promoting tissue repair or regeneration in a subject requiring promotion of tissue repair or regeneration, comprising the step of contacting the subject tissue with a self-assembling peptide or pharmaceutical composition described herein to thereby promote tissue repair or regeneration of the tissue. In some embodiments, the tissue is skin, bone, cartilage, nerve tissue, ligaments, tendons, vascular tissue, or muscle. In some embodiments, the tissue is eye tissue. In some embodiments, the tissue is cardiac tissue. In some embodiments, the subject has a congenital disease or disorder that necessitates the need for tissue repair or regeneration. In some embodiments, the subject suffers from an injury (e.g., surgery, trauma, stroke, tumor, or disease or disorder) that necessitates the need for tissue repair or regeneration.

[0025] In another embodiment, the present disclosure provides a method for promoting wound healing in a subject requiring accelerated wound healing, comprising the steps of contacting (or, instead administering to) the subject wound with an effective amount of a self-assembling peptide or pharmaceutical composition described herein, thereby promoting wound healing and / or antimicrobial activity. In some embodiments, the wound includes or comprises abrasions, burns, cracks, crushes, cuts, ulcers, lacerations, incisions, or scratches.

[0026] In another aspect, the Disclosure provides a method for stopping or preventing (or, instead, reducing) bleeding at a site within a subject, comprising the steps of bringing the site into contact with (or, instead, administering to a wound in the subject) a self-assembling peptide or pharmaceutical composition described herein, thereby creating a physical barrier, and thereby stopping or preventing (or reducing) bleeding at the site within the subject.

[0027] In another embodiment, the present disclosure provides a method for excising a lesion from a site of the gastrointestinal tract of interest, comprising the steps of: contacting (or, instead, administering to the submucosa beneath the lesion) the submucosa beneath the lesion with an amount sufficient to lift the lesion; and excising the lesion from a site of the gastrointestinal tract of interest. In some embodiments, the lesion includes polyps, ulcers, or tumors. In some embodiments, the lesion is located in a region of the gastrointestinal tract of interest selected from the mouth, throat, esophagus, stomach, small intestine, large intestine, colon, and rectum.

[0028] In yet another aspect, the disclosure provides a method for culturing cells, comprising the step of bringing the cells into contact with a pharmaceutical composition described herein.

[0029] In another embodiment, a method for treating an intraalveolar cyst of the target, The steps include: introducing the delivery device into the target region of the target alveolar cyst; The steps include positioning the end of the delivery device in the target region where treatment of alveolar cysts is desired; The procedure involves administering an effective amount and effective concentration of the self-assembling peptide or pharmaceutical composition described herein to a target region through a delivery device to treat alveolar cysts by forming a barrier under physiological conditions in the target region; The steps include removing the delivery device from the target region; A step to disintegrate alveolar cysts before or after administration of the solution. Methods including the above are provided herein.

[0030] In another embodiment, a method for reducing adhesion to biological tissue is provided herein, comprising the step of administering an effective amount of a self-assembling peptide or pharmaceutical composition described herein to a biological tissue, thereby reducing adhesion to the biological tissue.

[0031] In another embodiment, a method for filling the interosseous space of a target bone, Steps include introducing the delivery device into the target bone; The steps include positioning the end of the delivery device proximal to the interstitial space of the bone where bone growth promotion is desired; The procedure involves administering, through a delivery device, a self-assembling peptide or pharmaceutical composition described herein at a concentration sufficient to form a hydrogel scaffold under physiological conditions; Steps to remove the delivery device and Methods including the above are provided herein.

[0032] In another embodiment, a method for treating a target dry eye is provided herein, comprising the step of administering an effective amount of a self-assembling peptide or pharmaceutical composition described herein to the target eye.

[0033] The present invention further provides self-assembling peptides comprising or consisting of the following amino acid sequences for use in any of the methods described herein: [(X)i(Y)j(Z)k(Y)l]m(X)n (Formula I), [(Y)i(X)j(Y)k(Z)l]m(Y)n (Formula II), [(Z)i(Y)j(X)k(Y)l]m(Z)n (Formula III), or [(Y)i(Z)j(Y)k(X)l]m(Y)n (Formula IV).

[0034] Other features and advantages of the present invention will become apparent from the following detailed description and drawings, as well as from the claims. In embodiments of the present invention, for example, the following items are provided. (Item 1) The amino acid sequence shown below: [(X)i(Y)j(Z)k(Y)l]m(X)n Formula I, [(Y)i(X)j(Y)k(Z)l]m(Y)n Formula II, [(Z)i(Y)j(X)k(Y)l]m(Z)n Equation III, or [(Y)i(Z)j(Y)k(X)l]m(Y)n Equation IV (In the formula, each (X) is independently an ionic polar amino acid, each (Y) is independently a hydrophobic amino acid, each (Z) is independently a nonionic polar amino acid, each i, j, k, and l is independently an integer ≥ 1, m is an integer ≥ 2, and n = 0 or an integer ≥ 1) A pharmaceutical composition containing a self-assembling peptide. (Item 2) The pharmaceutical composition according to item 1, wherein the self-assembling peptide comprises the amino acid sequence shown in formula I. (Item 3) The pharmaceutical composition according to item 1, wherein the self-assembling peptide comprises the amino acid sequence shown in formula II. (Item 4) The pharmaceutical composition according to item 1, wherein the self-assembling peptide comprises the amino acid sequence shown in formula III. (Item 5) The pharmaceutical composition according to item 1, wherein the self-assembling peptide comprises the amino acid sequence shown in formula IV. (Item 6) A pharmaceutical composition according to any one of items 1 to 5, wherein each (X) is a basic amino acid selected from the group consisting of arginine, lysine, histidine, and ornithine. (Item 7) A peptide according to any one of items 1 to 5, wherein each (X) is an acidic amino acid selected from the group consisting of aspartic acid and glutamic acid. (Item 8) A pharmaceutical composition according to any one of items 1 to 7, wherein each (Y) is selected from the group consisting of alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, and glycine. (Item 9) A pharmaceutical composition according to any one of items 1 to 8, wherein each (Z) is selected from the group consisting of serine, threonine, tyrosine, cysteine, glutamine, asparagine, and methionine. (Item 10) A pharmaceutical composition according to any one of items 1 to 9, wherein each of i, j, k, and l is 1. (Item 11) A pharmaceutical composition according to any one of items 1 to 10, wherein m is an integer 2, 3, or 4. (Item 12) The pharmaceutical composition according to any one of items 1 to 11, wherein the self-assembling peptide comprises the amino acid sequences shown in SEQ ID NOs: 1 to 20 and 94 to 96. (Item 13) The self-assembling peptide is N-terminal functional groups selected from the group consisting of acetyl, formyl, pyroglutamyl (pGlu), biotin, polyethylene glycol (PEG), urea, alkylamine, carbamate, sulfonamide, dansyl, 2,4-dinitrophenyl, fluorescein, 7-methoxycoumarin acetate, 9-fluorenylmethyloxycarbonyl, palmitic acid, succinyl, chloroacetyl, maleimide, benzyloxycarbonyl, bromoacetyl, nitrilotriacetyl, tert-butoxycarbonyl, 4-hydroxyphenylpropionic acid, allyloxycarbonyl, butyric acid, fatty acids, and trityl, and C-terminal functional groups selected from the group consisting of amines, amides, N-alkylamides, aldehydes, esters, alcohols, para-nitroanilides (pNA), 7-amino-4-methylcoumarins (Amc), hydrazides, hydroxamic acids, chloromethyl ketones, p-nitroanilines, para-nitrophenols, hydroxysuccinimide esters, fluoromethyl ketones, cysteamides, 9-fluorenemethyl (Fm) esters, allyl esters, 2,4-dimethoxybenzyl esters, 2-phenylisopropyl esters, p-nitrobenzyl esters, and 2-chlorotrityl esters. A pharmaceutical composition according to any one of items 1 to 12, comprising one or both of the above. (Item 14) The pharmaceutical composition according to any one of items 1 to 13, wherein the self-assembling peptide comprises the amino acid sequences shown in SEQ ID NOs. 21-40 and 97-99. (Item 15) The pharmaceutical composition according to any one of items 1 to 14, wherein the self-assembling peptide further comprises at least one biologically active peptide motif at the N-terminus, C-terminus, or both of the self-assembling peptide. (Item 16) The pharmaceutical composition according to item 15, 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. (Item 17) The pharmaceutical composition according to item 15 or 16, wherein the self-assembling peptide comprises the amino acid sequence shown in SEQ ID NOs. 71 to 90. (Item 18) One or more salts selected from the group consisting of NaCl, KCl, MgCl2, CaCl2, NH4Cl, Na2HPO4, KH2PO4, and CaSO4; or One or more sugars selected from the group consisting of dextrose, mannitol, glycerol, sucrose, and trehalose. A pharmaceutical composition according to any one of items 1 to 17, further comprising an isotonic agent containing the above. (Item 19) The pharmaceutical composition according to item 18, wherein the isotonic agent comprises one or more salts and is present at a concentration of about 0.01 M to about 0.3 M or about 0.15 M. (Item 20) The pharmaceutical composition according to item 18, wherein the isotonic agent contains 1 or more sugars and is present in a concentration of about 0.1 to 10% (w / v) or about 10% (w / v). (Item 21) A pharmaceutical composition according to any one of items 1 to 20, having a pH of approximately 6 to approximately 8. (Item 22) The pharmaceutical composition according to item 21, wherein the net charge of the self-assembling peptide is greater than or equal to +1 or less than or equal to -1. (Item 23) A pharmaceutical composition according to any one of items 1 to 22, wherein the concentration of the self-assembling peptide is approximately 0.01% (w / v) to approximately 10% (w / v), approximately 0.1% (w / v) to approximately 5% (w / v), approximately 0.5% (w / v) to approximately 1.5% (w / v), or approximately 1% (w / v). (Item 24) A pharmaceutical composition according to any one of items 1 to 23, further comprising isolated cells. (Item 25) The pharmaceutical composition according to item 24, wherein the isolated cells are mammalian cells selected from the group consisting of immune cells, stem cells, chondrocyte progenitor cells, pancreatic progenitor cells, myoblasts, fibroblasts, keratinocytes, nerve cells, glial cells, astrocytes, preadipocytes, adipocytes, vascular endothelial cells, endothelial progenitor cells, mesenchymal cells, neural stem cells, immune cells (e.g., B cells and T cells), smooth muscle progenitor cells, cardiomyocytes, fetal dermal fibroblasts, epidermal keratinocytes, myoblasts, and capillary endothelial cells. (Item 26) A pharmaceutical composition according to any one of items 1 to 25, further comprising a bioactive agent. (Item 27) The pharmaceutical composition according to item 26, wherein the bioactive agent is selected from the group consisting of hormones, growth factors, insulin, enzymes, siRNA, shRNA, antisense RNA, antibiotics, antibodies, and anti-inflammatory agents. (Item 28) A pharmaceutical composition, which is an aqueous solution, as described in any one of items 1 to 27. (Item 29) A pharmaceutical composition according to any one of items 1 to 27, which is a hydrogel. (Item 30) The pharmaceutical composition according to item 29, wherein the hydrogel has a storage modulus of at least about 10 pascals (Pa). (Item 31) A manufactured article comprising a pharmaceutical composition as described in any one of items 1 to 30, which is a syringe, vial, autoinjector, tube, or catheter. (Item 32) A method for promoting tissue repair or regeneration in a subject requiring promotion of tissue repair or regeneration, comprising the step of administering an effective amount of any one of items 1 to 30 to the subject tissue, thereby promoting tissue repair or regeneration of the subject tissue. (Item 33) The method according to item 32, wherein the tissue is skin, bone, cartilage, nerve, ligament, tendon, vascular tissue, eye, muscle, or heart tissue. (Item 34) The method according to item 32 or 33, wherein the subject has a congenital disorder or disability that necessitates tissue repair or regeneration; or the subject has an injury that necessitates tissue repair or regeneration, and the injury is the result of surgery, trauma, stroke, tumor, or disease or disability. (Item 35) A method for promoting wound healing in a subject requiring the promotion of wound healing, comprising the step of administering an effective amount of any one of items 1 to 30 to a wound in a subject, thereby promoting wound healing and / or antimicrobial activity, wherein the wound includes abrasions, burns, cracks, crushes, cuts, ulcers, lacerations, incisions or scratches. (Item 36) A method for reducing bleeding at a site within a subject, comprising the step of administering an effective amount of a pharmaceutical composition described in any one of items 1 to 30 to the site, wherein the pharmaceutical composition creates a physical barrier, thereby reducing bleeding at the site within the subject. (Item 37) A method of removing a lesion from the target gastrointestinal tract, The step of administering a sufficient amount of the pharmaceutical composition described in any one of items 1 to 30 to lift the lesion into the submucosa beneath the lesion; The steps of removing the lesion from the aforementioned part of the gastrointestinal tract of the subject and A method that includes this. (Item 38) The method according to item 37, wherein the lesion includes a polyp, ulcer, or tumor. (Item 39) The method according to item 37 or 38, wherein the lesion is located in a region of the gastrointestinal tract selected from the group consisting of the mouth, throat, esophagus, stomach, small intestine, large intestine, colon, and rectum. (Item 40) A method for culturing cells, comprising the step of contacting the cells with a pharmaceutical composition described in any one of items 1 to 30. (Item 41) A method for treating target alveolar cysts, The steps include introducing the delivery device into the target region of the alveolar cyst of the target; The steps include positioning the end of the delivery device in the target region where treatment of the alveolar cysts is desired; The steps include administering the pharmaceutical composition according to any one of items 1 to 30 for effective amount and effective concentration to the target region through the delivery device to treat the alveolar cysts by forming a barrier under physiological conditions in the target region; The steps include removing the delivery device from the target region; A step of disintegrating the alveolar cysts before or after administration of the solution. A method that includes this. (Item 42) A method for reducing adhesion to a biological tissue, comprising the step of administering an effective amount of a pharmaceutical composition described in any one of items 1 to 30 to the biological tissue, thereby reducing adhesion to the biological tissue. (Item 43) A method for filling the interosseous space in question, Steps include introducing the delivery device into the target bone; The steps include positioning the end of the delivery device proximal to the interstitial space of the bone where bone growth promotion is desired; The steps include administering, through the delivery device, a pharmaceutical composition according to any one of items 1 to 30 in a concentration sufficient to form a hydrogel scaffold under physiological conditions; The step of removing the delivery device A method that includes this. (Item 44) A method for treating a target dry eye, comprising the step of administering an effective amount of a pharmaceutical composition described in any one of items 1 to 30 to the target eye. (Item 45) The amino acid sequence shown below: [(X)i(Y)j(Z)k(Y)l]m(X)n Formula I, [(Y)i(X)j(Y)k(Z)l]m(Y)n Formula II, [(Z)i(Y)j(X)k(Y)l]m(Z)n Equation III, or [(Y)i(Z)j(Y)k(X)l]m(Y)n Equation IV (In the formula, each (X) is independently an ionic polar amino acid, each (Y) is independently a hydrophobic amino acid, each (Z) is independently a nonionic polar amino acid, each i, j, k, and l is independently an integer ≥ 1, m is an integer ≥ 2, and n = 0 or an integer ≥ 1) Self-assembling peptides containing these peptides. [Brief explanation of the drawing]

[0035] [Figure 1] Figure 1 shows net charge measurements of KLNL12 (SEQ ID NO: 21) at various pH levels.

[0036] [Figure 2] Figure 2 shows net charge measurements of NLEL12 (SEQ ID NO: 33) at various pH levels.

[0037] [Figure 3] Figure 3 shows net charge measurements of RADA16 (SEQ ID NO: 91) at various pH levels.

[0038] [Figure 4] Figure 4 shows a schematic diagram of the molecular structure and electronic charge of RANA16 (SEQ ID NO: 39) at pH 7.5.

[0039] [Figure 5] Figure 5 shows a schematic diagram of the molecular structure and electronic charge of RADA16 (SEQ ID NO: 91) at pH 7.5.

[0040] Figures 6 to 8 show the shear viscosity reduction properties of exemplary aqueous pharmaceutical compositions containing the self-assembling peptides KLNL12 (SEQ ID NO: 21), IQIK13 (SEQ ID NO: 28), or NLEL12 (SEQ ID NO: 33).

[0041] [Figure 6] Figure 6 shows the shear viscosity reduction properties of an aqueous pharmaceutical composition containing 1% (w / v) KLNL12 (SEQ ID NO: 21) at pH 7.5, as reflected by varying the viscosity according to various shear rates.

[0042] [Figure 7] Figure 7 shows the shear viscosity reduction properties of an aqueous pharmaceutical composition containing 1% (w / v) IQIK13 (SEQ ID NO: 28) at pH 7.5, as reflected by varying the viscosity according to various shear rates.

[0043] [Figure 8] Figure 8 shows the shear viscosity reduction properties of an aqueous pharmaceutical composition containing 1% (w / v) NLEL12 (SEQ ID NO: 33) at pH 7.5, as reflected by varying the viscosity according to various shear rates.

[0044] Figures 9-10 show the shear viscosity reduction properties of exemplary aqueous pharmaceutical compositions containing the self-assembling peptide KLNL12 (SEQ ID NO: 21) or NLEL12 (SEQ ID NO: 33).

[0045] [Figure 9] Figure 9 shows the shear viscosity reduction properties of an aqueous pharmaceutical composition containing 1% (w / v) KLNL12 (SEQ ID NO: 21) with 0.9% NaCl at pH 7.5, as reflected by varying the viscosity according to various shear rates.

[0046] [Figure 10] Figure 10 shows the shear viscosity reduction properties of an aqueous pharmaceutical composition containing 1% NLEL12 (SEQ ID NO: 33) with 0.9% NaCl at pH 7.5, as reflected by varying the viscosity according to various shear rates.

[0047] Figures 11 and 12 show the thixotropic properties of exemplary aqueous pharmaceutical compositions containing the self-assembling peptide KLNL12 (SEQ ID NO: 21) or NLEL12 (SEQ ID NO: 33).

[0048] [Figure 11] Figure 11 shows the thixotropic properties of an aqueous pharmaceutical composition containing 1% (w / v) KLNL12 (SEQ ID NO: 21) with 0.9% NaCl at pH 7.5, as reflected by the change in mechanical strength after the removal of shear stress.

[0049] [Figure 12] Figure 12 shows the thixotropic properties of an aqueous pharmaceutical composition containing 1% NLEL12 (SEQ ID NO: 33) with 0.9% NaCl at pH 7.5, as reflected by the change in mechanical strength after the removal of shear stress.

[0050] Figures 13 to 15 show the changes in the rheological properties of exemplary aqueous pharmaceutical compositions containing the self-assembling peptides KLNL12 (SEQ ID NO: 21), KIQI13 (SEQ ID NO: 29), or NLEL12 (SEQ ID NO: 33) in response to exposure to DMEM.

[0051] [Figure 13] Figure 13 shows the changes in the rheological properties of an aqueous pharmaceutical composition containing 1% (w / v) KLNL12 (SEQ ID NO: 21) at pH 7.5, as reflected by the increase in mechanical strength after exposure to DMEM.

[0052] [Figure 14] Figure 14 shows the changes in the rheological properties of an aqueous pharmaceutical composition containing 1% (w / v) KIQI13 (SEQ ID NO: 29) at pH 7.5, as reflected by the increase in mechanical strength after exposure to DMEM.

[0053] [Figure 15]Figure 15 shows the changes in the rheological properties of an aqueous pharmaceutical composition containing 1% (w / v) NLEL12 (SEQ ID NO: 33) at pH 7.5, as reflected by the increase in mechanical strength after exposure to DMEM.

[0054] Figures 16-18 show the changes in the rheological properties of exemplary aqueous pharmaceutical compositions containing the self-assembling peptides KLNL12 (SEQ ID NO: 21), KIQI13 (SEQ ID NO: 29), or NLEL12 (SEQ ID NO: 33) in response to exposure to DMEM.

[0055] [Figure 16] Figure 16 shows the changes in the rheological properties of an aqueous pharmaceutical composition containing 1% (w / v) KLNL12 (SEQ ID NO: 21) with 0.9% NaCl at pH 7.5, as reflected by the increase in mechanical strength after exposure to DMEM.

[0056] [Figure 17] Figure 17 shows the changes in the rheological properties of an aqueous pharmaceutical composition containing 1% (w / v) KIQI13 (SEQ ID NO: 29) with 0.9% NaCl at pH 7.5, as reflected by the increase in mechanical strength after exposure to DMEM.

[0057] [Figure 18] Figure 18 shows the changes in the rheological properties of an aqueous pharmaceutical composition containing 1% NLEL12 (SEQ ID NO: 33) with 0.9% NaCl at pH 7.5, as reflected by the increase in mechanical strength after exposure to DMEM.

[0058] [Figure 19] Figure 19 is a schematic diagram of a polymer structure that can be formed by the self-assembling peptide of the present invention.

[0059] [Figure 20] Figures 20A to 20C below are photographs showing the appearance of aqueous pharmaceutical compositions containing specific self-assembling peptides at pH 7.5. Figure 20A is a photograph showing the appearance of an aqueous pharmaceutical composition containing RADA16 (SEQ ID NO: 91) at pH 7.5.

[0060] Figure 20B is a photograph showing the appearance of an aqueous pharmaceutical composition containing KLNL12 (SEQ ID NO: 21) at pH 7.5.

[0061] Figure 20C is a photograph showing the appearance of an aqueous pharmaceutical composition containing NLEL12 (SEQ ID NO: 33) at pH 7.5.

[0062] [Figure 21] Figures 21A and 21B show the appearance of the submucosa of the pig stomach after injection of specific pharmaceutical compositions at 0–5 minutes (Figure 21A) and 15–20 minutes (Figure 21B). Injection sites are indicated by black dotted ellipses: (1) 2 mL of 0.1% (w / v) KLNL12 (SEQ ID NO: 21), 0.9% (w / v) NaCl pH 7.5; (2) 2 mL of 0.1% (w / v) NLKL12 (SEQ ID NO: 23), 0.9% (w / v) NaCl pH 7.5; (3) 2 mL of 0.1% (w / v) KIQI13 (SEQ ID NO: 29), 0.9% (w / v) NaCl pH 7.5; (4) 2 mL of 0.2% (w / v) RADA16 (SEQ ID NO: 91) pH 2.5; (5) 2 mL of MucoUp®; (6) 2 mL of physiological saline pH 7.5.

[0063] [Figure 22] Figures 22A to 22C show the appearance of the submucosa of a pig's stomach after injection of either 2 mL of pH 7.5 physiological saline (Figure 22A), an aqueous pharmaceutical composition containing 0.2% (w / v) RADA16 (SEQ ID NO: 91) at pH 2.5 (Figure 22B), or an aqueous pharmaceutical composition containing 0.2% (w / v) RADA16 (SEQ ID NO: 91) and 0.9% NaCl (w / v) at pH 2.5 (Figure 22C). Black ovals indicate areas with a white, cloudy, and irregular appearance.

[0064] [Figure 23]Figures 23A, 23B, 23C, and 23D show the appearance of the submucosa of a pig's stomach after injection of a pharmaceutical composition containing 2 mL of pH 7.5 0.1% KLNL12 (SEQ ID NO: 21) and 0.9% NaCl (Figure 23A); a pharmaceutical composition containing 2 mL of pH 7.5 0.2% NLEL12 (SEQ ID NO: 33) and 0.9% NaCl (Figure 23B); a pharmaceutical composition containing 2 mL of pH 7.5 0.2% QLEL12 (SEQ ID NO: 35) and 0.9% NaCl (Figure 23C); or a pharmaceutical composition containing 2 mL of pH 7.5 0.2% LELQ12 (SEQ ID NO: 36) and 0.9% NaCl.

[0065] [Figure 24] Figure 24A shows the changes in the rheological properties of an aqueous pharmaceutical composition containing 0.15% (w / v) QLEL12 (SEQ ID NO: 35) with 0.9% NaCl (w / v) at pH 7.5, as reflected by the increase in mechanical strength after exposure to DMEM.

[0066] Figure 24B shows the rheological properties of aqueous pharmaceutical compositions containing QLEL12 (SEQ ID NO: 35) with 0.9% NaCl (w / v) at pH 7.5 at various concentrations from 0.1% (w / v) to 0.3% (w / v); linear regression was performed.

[0067] [Figure 25-1] Figures 25A to 25D show the elevation height and lifting capacity of the submucosa of the canine stomach and colon after injection of QLEL12 (SEQ ID NO: 35) containing 0.5 mL of 0.9% NaCl (w / v) at pH 7.5 in various concentrations from 0.1% (w / v) to 0.3% (w / v). [Figure 25-2] Figures 25A to 25D show the elevation height and lifting capacity of the submucosa of the canine stomach and colon after injection of QLEL12 (SEQ ID NO: 35) containing 0.5 mL of 0.9% NaCl (w / v) at pH 7.5 in various concentrations from 0.1% (w / v) to 0.3% (w / v). [Modes for carrying out the invention]

[0068] These and other advantages of this technology will become clear when you refer to the following explanation.

[0069] Detailed explanation definition Unless otherwise defined herein, scientific and technical terms used herein have the meanings generally understood by those skilled in the art. Generally, the nomenclature and techniques used herein in relation to chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics, and protein and nucleic acid chemistry are well known and commonly used in the art.

[0070] The methods and techniques described herein are generally carried out in accordance with the prior art methods well known in the art, unless otherwise indicated, and as described in the various general and more specific references cited and discussed throughout this specification.

[0071] The chemical terms used herein are used in accordance with their prior art usage, as exemplified in "The McGraw-Hill Dictionary of Chemical Terms," ​​edited by Parker S., McGraw-Hill, San Francisco, CA (1985).

[0072] All publications, patents, and published patent disclosures referenced in this disclosure are incorporated herein by reference. In case of any conflict, this specification, including specific definitions, shall prevail.

[0073] As used herein, the term “self-assembly” refers to the ability of certain peptides to spontaneously self-associate into higher-order structures (e.g., β-sheets). For example, a pharmaceutical composition containing self-assembling peptides in solution form transitions to a gel state when the self-assembling peptides self-associate. In some embodiments, the interactions between and within individual self-assembling peptides are reversible so that the composition can reversibly transition between a gel state and a solution state. The interactions between and within individual self-assembling peptides can be non-covalent interactions, including hydrogen bonds, ionic interactions, electrostatic interactions (e.g., via van der Waals forces), and hydrophobic interactions. In various embodiments, the self-assembling peptide nanostructure contains peptides in the form of β-sheets. The nanostructure can be nanofibers or networks of nanofibers. For illustrative purposes, Figure 19 shows the organization of self-assembling peptides into β-sheets 1901. These β-sheets can self-assemble into nanofibers 1902. Multiple nanofibers 1902 can self-assemble into a membrane network 1903. In some embodiments, the self-assembly of the peptides described herein into higher-order structures is reactive to one or more environmental triggers (e.g., one or more changes among pH, temperature, ionic strength, osmotic pressure, applied pressure, applied shear stress, etc.).

[0074] As used herein, the term “administer” is intended to include, but is not limited to, administering, introducing, or injecting the self-assembling peptides and / or pharmaceutical compositions comprising the self-assembling peptides described herein.

[0075] As used herein, the terms “amino acid residue” or “amino acid” include natural and synthetic amino acid residues, including D- and L-amino acids; alpha-, beta- and gamma-amino acids; chemically modified amino acids; naturally occurring non-proteinogenic amino acids; rare amino acids; and chemically synthesized compounds having properties known in the art to be characteristic of amino acids.

[0076] As used herein, the phrases “therapeutic dose,” “effective dose,” or “effective amount” refer to the amount of a pharmaceutical composition that, when delivered to a subject, provides a therapeutic or aesthetic benefit in the treatment, prevention, or management of a tissue, disease, and / or disorder. Determining the therapeutic dose is within the scope of the skills of those skilled in the art. Generally, the therapeutic dose may vary depending on the subject’s medical history, age, symptoms, sex, and the severity and type of the subject’s condition, as well as the administration of other pharmaceutically active agents.

[0077] As used herein, the term "hydrogel" refers to a composition comprising a three-dimensional network of self-assembling peptides. The term hydrogel may be used to refer to a network of self-assembling peptides in a dry (xerogel) or wet state. In a wet state, hydrogels may contain a high water content (e.g., about 90% to about 99.9% water). Hydrogels possess many properties desirable for biomedical applications. For example, hydrogels can be manufactured to be non-toxic and compatible with tissues. Furthermore, hydrogels are typically highly permeable to water, ions, and small molecules.

[0078] As used herein, the term “isolated” with respect to cells refers to cells that have been mechanically separated from their natural environment.

[0079] As used herein, the term “isotonic agent” refers to an agent that may be used to adjust the osmotic pressure of the pharmaceutical compositions described herein. Exemplary isotonic agents include, but are not limited to, sodium chloride, calcium chloride, potassium chloride, potassium phosphate, and sugars (e.g., dextrose and sucrose). In some embodiments, the isotonic agent is sodium chloride. Isotonic agents can increase the rheological properties of solutions or compositions (e.g., hydrogels) containing the self-assembling peptides disclosed herein.

[0080] As used herein, “pharmaceutical composition” refers to a composition comprising a self-assembling peptide and other components such as a physiologically appropriate carrier and / or excipient.

[0081] As used herein, the terms “protein” and “peptide” are used interchangeably to refer to polymers of amino acid residues linked to one another by peptide bonds between the alpha-amino and carboxyl groups of adjacent residues. The terms “protein” and “peptide” include polymers having modified amino acid residues (e.g., phosphorylated, amidated, or glycated amino acid residues) and amino acid analogs.

[0082] As used herein, the term “Subject” refers to either a human or a non-human animal. This term includes, but is not limited to, mammals (e.g., humans, other primates, pigs, rodents (e.g., mice and rats or hamsters), rabbits, guinea pigs, cattle, horses, cats, dogs, sheep and goats). In some embodiments, the subject is human. In some embodiments, the subject is an adult human subject. In some embodiments, the subject is a child human subject.

[0083] As used herein, the term “wound” refers to trauma to the tissue of an object (e.g., a human object), such as abrasions, burns, cracks, crushes, cuts, ulcers, lacerations, incisions, or scratches.

[0084] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention pertains. Similar or equivalent methods and materials may be used in carrying out or testing the present invention, but suitable methods and materials are listed below. All publications, patent applications, patents and other references referenced herein are incorporated by reference in their entirety. In case of any conflict, this specification, including definitions, shall prevail. Furthermore, materials, methods and examples are illustrative and not intended to limit the present invention. No citation or specification of any document in this application constitutes an endorsement that such document is available as prior art to the present invention.

[0085] Other features and advantages of the present invention will become apparent from the following detailed description and claims. Pharmaceutical compositions and self-assembling peptides

[0086] In one embodiment, the disclosure provides a self-assembling peptide as described herein and a pharmaceutical composition comprising at least one self-assembling peptide. When present in aqueous solution, the self-assembling peptides spontaneously assemble into higher-order structures via intermolecular and intramolecular electrostatic interactions. These higher-order structures include β-sheets, nanofiber structures, and three-dimensional network or mesh structures. When these higher-order structures form an aqueous solution, they can become a hydrogel with a variety of desirable properties. In addition to the self-assembling peptides, the pharmaceutical composition may also include other additives such as isotonic agents, buffers, pharmaceutically acceptable excipients, biomolecules, therapeutic agents, and cells.

[0087] In some embodiments, the self-assembling peptide is biodegradable. As used herein, “biodegradable” refers to a material that, in interaction with a physiological environment, decomposes or disintegrates into components that can be metabolized or excreted by the subject over a period ranging from minutes to years (e.g., 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 2 months, 3 months, 6 months, 1 year, 2 years, or more). In some embodiments, the self-assembling peptide may be decomposed by cleavage of the peptide chain, for example, via hydrolysis or enzymatic cleavage. While the self-assembling peptide (and the higher-order structures formed by the self-assembling peptide, e.g., scaffolds)) may be biodegradable, these higher-order structures preferably maintain their structural integrity for the period required for their intended use.

[0088] The self-assembling peptides provided herein have the following amino acid sequence [(X)i(Y)j(Z)k(Y)l]m(X)n (Formula I), [(Y)i(X)j(Y)k(Z)l]m(Y)n (Formula II), [(Z)i(Y)j(X)k(Y)l]m(Z)n(Equation III), or [(Y)i(Z)j(Y)k(X)l]m(Y)n (Formula IV) (In the formula, each (X) is independently an ionic polar amino acid, each (Y) is independently a hydrophobic amino acid, each (Z) is independently a nonionic polar amino acid, each i, j, k, and l is independently an integer ≥ 1, m is an integer ≥ 2, and n = 0 or an integer ≥ 1) It includes or consists of. In some embodiments, each i, j, k and l is independently an integer 1. In some embodiments, each i, j, k and l is independently an integer 2. In some embodiments, each i, j, k and l is independently an integer 3. In some embodiments, n is 0. In some embodiments, n is an integer ≥ 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 1. In some embodiments, m is an integer ≥ 2 (e.g., 2, 3, 4, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 2, 3 or 4. In some embodiments, each (X) is the same amino acid, each (Y) is the same amino acid, and / or each (Z) is an amino acid residue of the same type.

[0089] In some embodiments, the self-assembling peptide comprises or consists of the amino acid sequence shown in [(X)i(Y)j(Z)k(Y)l]m(X)n(Formula I) (wherein each (X) is independently an ionic polar amino acid, each (Y) is independently a hydrophobic amino acid, each (Z) is independently a nonionic polar amino acid, each i, j, k and l is independently an integer ≥ 1, m is an integer ≥ 2, and n = 0 or an integer ≥ 1). In some embodiments, each i, j, k and l is independently an integer 1. In some embodiments, each i, j, k and l is independently an integer 2. In some embodiments, each i, j, k and l is independently an integer 3. In some embodiments, n is 0. In some embodiments, n is an integer ≥ 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 1. In some embodiments, m is an integer ≥ 2 (e.g., 2, 3, 4, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 2, 3, or 4.

[0090] In some embodiments, the self-assembling peptide comprises or consists of the amino acid sequence shown in [(Y)i(X)j(Y)k(Z)l]m(Y)n(Formula II) (wherein each (X) is independently an ionic polar amino acid, each (Y) is independently a hydrophobic amino acid, each (Z) is independently a nonionic polar amino acid, each i, j, k and l is independently an integer ≥ 1, m is an integer ≥ 2, and n = 0 or an integer ≥ 1). In some embodiments, each i, j, k and l is independently an integer 1. In some embodiments, each i, j, k and l is independently an integer 2. In some embodiments, each i, j, k and l is independently an integer 3. In some embodiments, n is 0. In some embodiments, n is an integer ≥ 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 1. In some embodiments, m is an integer ≥ 2 (e.g., 2, 3, 4, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 2, 3, or 4.

[0091] In some embodiments, the self-assembling peptide comprises or consists of the amino acid sequence shown in [(Z)i(Y)j(X)k(Y)l]m(Z)n(Formula III) (wherein each (X) is independently an ionic polar amino acid, each (Y) is independently a hydrophobic amino acid, each (Z) is independently a nonionic polar amino acid, each i, j, k and l is independently an integer ≥ 1, m is an integer ≥ 2, and n = 0 or an integer ≥ 1). In some embodiments, each i, j, k and l is independently an integer 1. In some embodiments, each i, j, k and l is independently an integer 2. In some embodiments, each i, j, k and l is independently an integer 3. In some embodiments, n is 0. In some embodiments, n is an integer ≥ 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 1. In some embodiments, m is an integer ≥ 2 (e.g., 2, 3, 4, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 2, 3, or 4.

[0092] In some embodiments, the self-assembling peptide comprises or consists of the amino acid sequence shown in [(Y)i(Z)j(Y)k(X)l]m(Y)n(Formula IV) (wherein each (X) is independently an ionic polar amino acid, each (Y) is independently a hydrophobic amino acid, each (Z) is independently a nonionic polar amino acid, each i, j, k and l is independently an integer ≥ 1, m is an integer ≥ 2, and n = 0 or an integer ≥ 1). In some embodiments, each i, j, k and l is independently an integer 1. In some embodiments, each i, j, k and l is independently an integer 2. In some embodiments, each i, j, k and l is independently an integer 3. In some embodiments, n is 0. In some embodiments, n is an integer ≥ 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 1. In some embodiments, m is an integer ≥ 2 (e.g., 2, 3, 4, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 2, 3, or 4.

[0093] In some embodiments, the self-assembling peptide comprises or consists of an amino acid sequence represented by [(X)i(Y)j(Z)k(Y)l]m(Formula V) (wherein each (X) is independently an ionic polar amino acid, each (Y) is independently a hydrophobic amino acid, each (Z) is independently a nonionic polar amino acid, each i, j, k, and l is independently an integer ≥ 1, m is an integer ≥ 2, and n = 0 or an integer ≥ 1). In some embodiments, each i, j, k, and l is independently an integer 1. In some embodiments, each i, j, k, and l is independently an integer 2. In some embodiments, each i, j, k, and l is independently an integer 3. In some embodiments, n is 0. In some embodiments, n is an integer ≥ 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 1. In some embodiments, m is an integer ≥ 2 (e.g., 2, 3, 4, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 2, 3, or 4.

[0094] In some embodiments, the self-assembling peptide comprises or consists of the amino acid sequence shown in [(Y)i(X)j(Y)k(Z)l]m (Formula VI) (wherein each (X) is independently an ionic polar amino acid, each (Y) is independently a hydrophobic amino acid, each (Z) is independently a nonionic polar amino acid, each i, j, k and l is independently an integer ≥ 1, m is an integer ≥ 2, and n = 0 or an integer ≥ 1). In some embodiments, each i, j, k and l is independently an integer 1. In some embodiments, each i, j, k and l is independently an integer 2. In some embodiments, each i, j, k and l is independently an integer 3. In some embodiments, n is 0. In some embodiments, n is an integer ≥ 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 1. In some embodiments, m is an integer ≥ 2 (e.g., 2, 3, 4, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 2, 3, or 4.

[0095] In some embodiments, the self-assembling peptide comprises or consists of the amino acid sequence shown in [(Z)i(Y)j(X)k(Y)l]m (Formula VIII) (wherein each (X) is independently an ionic polar amino acid, each (Y) is independently a hydrophobic amino acid, each (Z) is independently a nonionic polar amino acid, each i, j, k, and l is independently an integer ≥ 1, m is an integer ≥ 2, and n = 0 or an integer ≥ 1). In some embodiments, each i, j, k, and l is independently an integer 1. In some embodiments, each i, j, k, and l is independently an integer 2. In some embodiments, each i, j, k, and l is independently an integer 3. In some embodiments, n is 0. In some embodiments, n is an integer ≥ 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 1. In some embodiments, m is an integer ≥ 2 (e.g., 2, 3, 4, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 2, 3, or 4.

[0096] In some embodiments, the self-assembling peptide comprises or consists of the amino acid sequence shown in [(Y)i(Z)j(Y)k(X)l]m (Formula VIII) (wherein each (X) is independently an ionic polar amino acid, each (Y) is independently a hydrophobic amino acid, each (Z) is independently a nonionic polar amino acid, each i, j, k, and l is independently an integer ≥ 1, m is an integer ≥ 2, and n = 0 or an integer ≥ 1). In some embodiments, each i, j, k, and l is independently an integer 1. In some embodiments, each i, j, k, and l is independently an integer 2. In some embodiments, each i, j, k, and l is independently an integer 3. In some embodiments, n is 0. In some embodiments, n is an integer ≥ 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 1. In some embodiments, m is an integer ≥ 2 (e.g., 2, 3, 4, 6, 7, 8, 9, 10, 11, 12 or greater). In some embodiments, m is an integer 2, 3, or 4.

[0097] Each (X) in the self-assembling peptides of formulas I to IV may be an acidic amino acid or a basic amino acid. For example, in some embodiments, each (X) is a basic amino acid, but is not limited to arginine, lysine, histidine, and ornithine. In some embodiments, each (X) is an acidic amino acid, but is not limited to aspartic acid and glutamic acid.

[0098] Each (Y) in the self-assembling peptides of formulas I-IV may be alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, or glycine.

[0099] Each (Z) is selected from the group consisting of serine, threonine, tyrosine, cysteine, glutamine, asparagine, and methionine.

[0100] The pharmaceutical compositions described herein may comprise one type of self-assembling peptide or several different types of self-assembling peptides. For example, in some embodiments, the pharmaceutical compositions provided herein may comprise one type of self-assembling peptide (e.g., a self-assembling peptide comprising or consisting of the amino acid sequence shown in formula I). ​​In some embodiments, the pharmaceutical compositions described herein may comprise two or more types of self-assembling peptides (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10 or more types). If the pharmaceutical compositions provided herein comprise more than one type of self-assembling peptide, all self-assembling peptides may comprise an amino acid sequence represented by a single formula (i.e., one of formulas I to IV) or an amino acid sequence represented by different formulas. When multiple types of self-assembling peptides are present in the pharmaceutical compositions provided herein, different types of peptides may interact and form higher-order structures (e.g., β-sheets).

[0101] In some embodiments, the self-assembling peptide contains or consists of approximately 8 to 50 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of approximately 8 to 18 amino acid residues. For example, the self-assembling peptide may contain 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 40 amino acid residues. The size of the self-assembling peptide may be such that the tertiary structure of the self-assembling peptide does not disrupt the peptide's ability to form higher-order structures (e.g., nanofibers or β-sheets) with other self-assembling peptides. In some embodiments, the self-assembling peptide contains or consists of about 12 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 13 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 14 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 15 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 16 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 17 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 18 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 19 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 20 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 21 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 22 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 23 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 24 amino acid residues.In some embodiments, the self-assembling peptide contains or consists of about 25 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 26 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 27 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 28 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 29 amino acid residues. In some embodiments, the self-assembling peptide contains or consists of about 30 amino acid residues.

[0102] Exemplary self-assembling peptides are provided below in Table 1. The exemplary self-assembling peptides in Table 1 include alternating ionic polar amino acid residues, hydrophobic amino acid residues, and nonionic polar amino acid residues. In some embodiments, the pharmaceutical compositions provided herein include self-assembling peptides comprising or consisting of the amino acid sequences shown in SEQ ID NOs: 1-20 and 94-96. Table 1. Exemplary self-assembling peptides [Table 1-1] [Table 1-2]

[0103] In some embodiments, the self-assembling peptide may contain one or more functional groups. In some embodiments, the functional groups prevent or delay the degradation of the self-assembling peptide. In some embodiments, the self-assembling peptide contains an N-terminal functional group. In some embodiments, the self-assembling peptide contains a C-terminal functional group. In some embodiments, the self-assembling peptide contains both an N-terminal and a C-terminal functional group. For example, in some embodiments, the functional groups prevent or delay the degradation of the self-assembling peptide by enzymes (e.g., in vivo) (e.g., acetyl, formyl, chloroacetyl, benzyloxycarbonyl, bromoacetyl, tert-butoxycarbonyl, 9-fluorenylmethyloxycarbonyl, allyloxycarbonyl, methyl ester, benzyl ester, or t-butyl ester). Examples of N-terminal functional groups include, but are not limited to, acetyl, formyl, pyroglutamyl (pGlu), biotin, polyethylene glycol (PEG), urea, alkylamines, carbamates, sulfonamides (e.g., 4-toluenesulfonyl, 4-nitrobenzenesulfonyl), dansyl, 2,4-dinitrophenyl (2,4-dintrophenyl), fluorescein, 7-methoxycoumarin acetate, 9-fluorenylmethyloxycarbonyl, palmitic acid, succinyl, chloroacetyl, maleimide, benzyloxycarbonyl, bromoacetyl, nitrilotriacetyl, tert-butoxycarbonyl, 4-hydroxyphenylpropionic acid, allyloxycarbonyl, butyric acid, fatty acids (e.g., hexanoic acid, octanoic acid, decanoic acid, palmitic acid, stearic acid, myristic acid, and lauric acid), and trityl.Examples of C-terminal functional groups include, but are not limited to, amides, N-alkylamides, aldehydes, esters (e.g., methyl esters, benzyl esters, or t-butyl esters), alcohols, para-nitroanilides (pNA), 7-amino-4-methylcoumarin (Amc), hydrazides, hydroxamic acids, chloromethyl ketones, p-nitroanilines, para-nitrophenols, hydroxysucinimide esters, fluoromethyl ketones, cysteamides, 9-fluorenemethyl (Fm) esters, allyl esters, 2,4-dimethoxybenzyl esters, 2-phenylisopropyl esters, p-nitrobenzyl esters, and 2-chlorotrityl esters.

[0104] In some embodiments, the self-assembling peptides described herein may include an N-terminal acetyl group and / or a C-terminal amine group. For example, exemplary self-assembling peptides containing both of these functional groups are provided below in Table 2. In some embodiments, the pharmaceutical compositions provided herein include self-assembling peptides comprising or consisting of the amino acid sequences shown in SEQ ID NOs. 21-40 and 97-99. Table 2. Exemplary self-assembling peptides containing an N-terminal acetyl group and a C-terminal amine group. [Table 2]

[0105] In some embodiments, the self-assembling peptides described herein include at least one biologically active peptide motif. While not wishing to be bound by any particular theory, the biologically active peptide motif may facilitate one or more biological processes in, on, or around the pharmaceutical compositions described herein (e.g., hydrogels), including, for example, cell adhesion, differentiation, proliferation, recruitment and / or homing; neurite extension; and recruitment (e.g., as a binding site for biologically active molecules), retention and / or reconfiguration. The biologically active peptide motif may be located anywhere along the amino acid sequence of the self-assembling peptide and preferably does not interfere with the self-assembling peptide's ability to form higher-order structures (e.g., nanofibers or β-sheets) through intramolecular or intermolecular interactions with other self-assembling peptides. For example, in some embodiments, the biologically active peptide motif is located at the N-terminus of the self-assembling peptide. In some embodiments, the biologically active peptide motif is located at the C-terminus of the self-assembling peptide. Self-assembling peptides can contain one or more (e.g., one, two, three, four, five, six, seven, eight, or more) biologically active peptide motifs. If a self-assembling peptide contains more than one biologically active peptide motif, each motif may be of the same type or may be different. Exemplary biologically active peptide motifs may be derived from proteins such as laminin-1, collagen IV, fibronectin, elastin, bone marrow homing peptide 1, bone marrow homing peptide 2, or myelopeptides. Non-limiting examples of biologically active motifs are provided below in Table 3. In some embodiments, the self-assembling peptides provided herein contain biologically active peptide motifs comprising the amino acid sequences shown in SEQ ID NOs. 41-70.

[0106] Table 3. Exemplary biologically active peptide motifs. [Table 3-1] [Table 3-2]

[0107] In some embodiments, the self-assembling peptide comprises a biologically active peptide motif as shown below in Table 4. In some embodiments, the self-assembling peptide comprises or consists of the amino acid sequences shown in SEQ ID NOs. 71-90.

[0108] Table 4. Exemplary self-assembling peptides containing biologically active peptide motifs. [Table 4]

[0109] The concentration of self-assembling peptides present in the pharmaceutical compositions provided herein can be varied to alter the rheological properties of the composition. In some embodiments, the pharmaceutical composition may contain about 0.01% (w / v) to about 10% (w / v) of the self-assembling peptides described herein. In some embodiments, the pharmaceutical composition may contain about 0.1% (w / v) to about 5% (w / v) of the self-assembling peptides described herein. In some embodiments, the pharmaceutical composition may contain about 0.1% (w / v) to about 2% (w / v) of the self-assembling peptides described herein. In some embodiments, the pharmaceutical composition may contain about 0.5% (w / v) to about 3% (w / v) of the self-assembling peptides described herein. In some embodiments, the pharmaceutical composition may contain about 0.5% (w / v) to about 1.5% (w / v) of the self-assembling peptides described herein. In some embodiments, the pharmaceutical composition may contain about 1% (w / v) to about 3% (w / v) of the self-assembling peptides described herein. In some embodiments, the pharmaceutical composition may contain about 0.5%, about 1% (w / v), about 1.5% (w / v), about 2% (w / v), about 2.5% (w / v), about 3% (w / v), about 3.5% (w / v), about 4% (w / v), about 4.5% (w / v), or about 5% (w / v) of the self-assembling peptides described herein.

[0110] In some embodiments, the pharmaceutical composition may be an aqueous solution or a hydrogel, or the pharmaceutical composition may be dehydrated (e.g., a powder). The hydrogels formed by the self-assembling peptides described herein may be porous or solid. In some embodiments, the hydrogels contain pores having an average diameter of about 1 nm to about 2000 μm, about 10 nm to about 1000 μm, about 10 nm to about 100 μm, about 10 nm to about 1 μm, about 5 nm to about 500 nm, or about 5 nm to about 250 nm.

[0111] In some embodiments, the pharmaceutical compositions provided herein may be hydrogels of different sizes and geometric shapes, comprising films and particles, such as nanoparticles or microparticles. In some embodiments, the pharmaceutical compositions may be laminated on a surface (e.g., a hydrogel). The particle size of the pharmaceutical composition will vary depending on the specific use intended for such particles. Generally, the particles may have at least one dimension in the range of about 1000 μm to about 2000 μm (e.g., about 1000 μm, about 1100 μm, 1200 μm, 1300 μm, 1400 μm, 1500 μm, 1600 μm, 1700 μm, 1800 μm, 1900 μm, or 2000 μm).

[0112] The self-assembling peptides described herein may be advantageously formulated at physiological pH, where the self-assembling peptides remain soluble and stable. In some embodiments, the pharmaceutical compositions described herein have a pH of about 5 to about 8. In some embodiments, the pharmaceutical compositions have a pH of about 6 to about 8. In some embodiments, the pharmaceutical compositions have a pH of about 5.5 to about 7.5. In some embodiments, the pharmaceutical compositions have a pH of about 6 to about 7.4. In some embodiments, the pharmaceutical compositions have a pH of about 6.5 to about 7.5. In some embodiments, the pharmaceutical compositions have a pH of about 7 to about 7.5. In some embodiments, the pharmaceutical compositions have a pH of about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, or about 8.0. The pH of the pharmaceutical compositions described herein may be adjusted by including a buffer system. Alternatively, self-assembling peptides may act as buffers in the pharmaceutical compositions. The pH of the pharmaceutical compositions may be adjusted using one or more acids or bases, such as sodium hydroxide, potassium hydroxide, hydrochloric acid, phosphoric acid, sodium carbonate, and sodium bicarbonate.

[0113] In some embodiments, the pharmaceutical composition may include a buffer such as phosphate-buffered saline (PBS). In some embodiments, the pharmaceutical composition may include, but is not limited to, a culture medium for cell culture, including Dulbecco's modified Eagle medium (DMEM), DME / F12, minimal essential medium (MEM), Eagle basal medium (BME), RPMI 1640, F-10, F-12, α-minimum essential medium (α-MEM), Glasgow minimal essential medium (G-MEM), PF CHO (SAFC Biosciences), and Iskov's modified Dulbecco medium.

[0114] The pharmaceutical compositions described herein can be formulated such that the composition has a neutral or physiological pH (e.g., pH about 6 to about 8) and the self-assembling peptides within it have a non-zero net charge. While we do not wish to be bound by any particular theory, when formulated at a neutral or physiological pH, the non-zero net charge of the self-assembling peptides allows the peptides to readily form electrostatic interactions with molecules of the opposite charge. This property can be advantageously used to promote adhesion of the pharmaceutical composition to a desired surface. This is particularly advantageous in clinical applications where the pharmaceutical composition is applied to tissues rich in biomolecules (e.g., glycoproteins) with a non-zero net charge.

[0115] Furthermore, while we do not wish to be bound by any particular theory, the self-assembling peptides described herein, which have a net positive charge when formulated at physiological pH, may be bactericidal and / or bacteriostatic, and are therefore particularly advantageous in therapeutic applications. Thus, self-assembling peptides can be readily used in a variety of applications where a reduction in bacterial load is desired.

[0116] In some embodiments, the net charge of the self-assembling peptide is greater than or equal to +1 or less than or equal to -1 when the pharmaceutical composition has a pH of about 5 to about 8. In some embodiments, the net charge of the self-assembling peptide is about +1 to about +6 (e.g., +1, +2, +3, +4, +5, or +6) when the pharmaceutical composition has a pH of about 5 to about 8 (e.g., 7.4). In some embodiments, the net charge of the self-assembling peptide is about -1 to about -6 (e.g., -1, -2, -3, -4, -5, or -6) when the pharmaceutical composition has a pH of about 5 to about 8 (e.g., 7.4).

[0117] In some embodiments, the pharmaceutical compositions provided herein have modulus of elasticity in the range of about 10 Pascals (Pa) to about 10,000 Pa. As used herein, the term “modulus of elasticity” refers to the tendency of a composition to deform elastically (i.e., non-permanently) when a force is applied to it. Generally, the modulus of elasticity of an object is defined as the slope of the stress-strain curve in the elastic deformation region. Many types of modulus of elasticity can be defined by specifying the method of measuring stress and strain, including the direction. Young's modulus (E) describes the tensile modulus, or the tendency of an object to deform along an axis when an opposing force is applied along the axis; it is defined as the ratio of tensile stress to tensile strain. This is usually simply called the modulus of elasticity. The shear modulus, or shear modulus (G or μ), describes the shear tendency of an object (deformation of shape in a given volume) when an opposing force is applied; it is defined as the shear stress relative to the shear strain. The shear modulus is a derivative of viscosity. The bulk modulus (K) is the volumetric modulus, or the tendency of an object to deform in all directions when a uniform load is applied in all directions; it is defined as the volumetric stress relative to the volumetric strain and is the reciprocal of the compressibility. The bulk modulus is a three-dimensional extension of Young's modulus. In some embodiments, the pharmaceutical compositions described herein (e.g., hydrogels) have a storage modulus of at least about 10 Pa. In some embodiments, the pharmaceutical compositions described herein have a storage modulus of at least about 50 Pa. In some embodiments, the pharmaceutical compositions described herein have a storage modulus of at least about 100 Pa. In some embodiments, the pharmaceutical compositions described herein have a storage modulus of at least about 500 Pa. In some embodiments, the pharmaceutical compositions described herein have a storage modulus of at least about 1000 Pa. In some embodiments, the pharmaceutical compositions described herein have a storage modulus of about 10 Pa to about 5000 Pa. In some embodiments, the pharmaceutical compositions described herein have a storage modulus of about 50 Pa to about 500 Pa. In some embodiments, the pharmaceutical compositions described herein have a storage modulus of about 10 Pa to about 1000 Pa. In some embodiments, the pharmaceutical compositions described herein have a storage modulus of about 500 Pa to about 5000 Pa.

[0118] In some embodiments, the increased ionic strength may increase the stiffness and / or gelation rate of self-assembling peptides present in the pharmaceutical compositions described herein. In some embodiments, the increased ionic strength may be physiological ionic strength. The ionic strength of a pharmaceutical composition may increase when the composition is administered into and / or onto a subject (e.g., a human subject). Alternatively, the ionic strength of a pharmaceutical composition described herein may increase by miscoating the composition with one or more isotonic agents. Exemplary isotonic agents include, but are not limited to, sodium chloride, calcium chloride, potassium chloride, and potassium phosphate. In some embodiments, the pharmaceutical compositions described herein do not contain an isotonic agent. In some embodiments, the pharmaceutical compositions described herein contain about 0.01 M to about 0.3 M of an isotonic agent. In some embodiments, the pharmaceutical compositions described herein contain about 0.1 M to about 0.3 M of an isotonic agent. In some embodiments, the pharmaceutical compositions described herein contain about 0.1 M to about 0.2 M of an isotonic agent. In some embodiments, the pharmaceutical compositions described herein include a 0.01M, 0.02M, 0.03M, 0.04M, 0.05M, 0.06M, 0.07M, 0.08M, 0.09M, 0.1M, 0.15M, 0.2M, 0.25M, or 0.3M isotonic agent.

[0119] In some embodiments, the pharmaceutical composition described herein is formulated to be hypotonic with respect to the target administration site. In some embodiments, the pharmaceutical composition described herein is formulated to be isotonic with respect to the target administration site. In some embodiments, the pharmaceutical composition described herein is formulated to be hypertonic with respect to the target administration site.

[0120] In some embodiments, the pharmaceutical composition described herein comprises cells (e.g., isolated cells). In some embodiments, the pharmaceutical composition (e.g., hydrogel) enhances the viability of cells therein, thereby facilitating the delivery of living cells to a desired site (e.g., damaged or defective body tissue). In some embodiments, the pharmaceutical composition comprises about 10 4 ~about 10 8 Cells / mL (e.g., 10) 4 pieces, 10 5 pieces, 10 6 pieces, 10 7 It is an aqueous solution containing (individual cells / mL). In some embodiments, the pharmaceutical composition is about 10 4 ~about 10 6 It is an aqueous solution containing individual cells / mL. In some embodiments, the pharmaceutical composition contains one cell type. In some embodiments, the pharmaceutical composition contains more than one cell type (e.g., two, three, four, five or more cell types).

[0121] Depending on the desired application, any suitable cells may be included in the pharmaceutical composition. For example, in some embodiments, animal or plant cells are seeded in the pharmaceutical composition (e.g., a hydrogel). In the case of a hydrogel, the cells may be encapsulated within the hydrogel. In some embodiments, the pharmaceutical composition includes mammalian cells. Exemplary mammalian cells include, but are not limited to, stem cells (embryonic stem cells, mesenchymal stem cells, bone marrow-derived stem cells, hematopoietic stem cells, neural stem cells, and hair follicle stem cells), chondrocyte progenitor cells, pancreatic progenitor cells, myoblasts, fibroblasts, keratinocytes, nerve cells, glial cells, astrocytes, preadipocytes, adipocytes, vascular endothelial cells, endothelial progenitor cells, mesenchymal cells, neural stem cells, immune cells (e.g., B cells and T cells), smooth muscle progenitor cells, cardiomyocytes, fetal skin fibroblasts, epidermal keratinocytes, myoblasts, and capillary endothelial cells. In some embodiments, the cells are genetically modified cells (for example, cells modified to express and secrete desired compounds such as growth factors, differentiation factors, cytokines, chimeric antigen receptors, or antibodies or fragments thereof). In some embodiments, the cells are tissue culture cell lines. Exemplary tissue culture cell lines include, but are not limited to, C166 cells, C6 glioma cell line, AML12, HeLa cells, and Chinese hamster ovary cells (CHO cells).

[0122] In some embodiments, the pharmaceutical compositions provided herein include, but are not limited to, bioactive molecules including extracellular matrix proteins (e.g., fibronectin, vitronectin, and laminin), cytokines, growth factors, differentiation factors, insulin, nucleic acids, vitamins, fatty acids, and therapeutic agents.

[0123] Examples of growth factors and cytokines include, but are not limited to, stem cell factor (SCF), granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage-stimulating factor (GM-CSF), stromal cell-derived factor-1, vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), angiopoietin, epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), hepatocyte nuclear factor-1 (HNF-1), nerve growth factor (NGF), bone morphogenetic protein (BMP), fibroblast growth factor (FGF), hepatocyte growth factor, insulin-like growth factor (IGF-1), interleukin (IL)-3, IL-1a, IL-1β, IL-6, IL-7, IL-8, IL-11 and IL-13, colony-stimulating factor, thrombopoietin, erythropoietin, fit3-ligand, and tumor necrosis factor α (TNFα).

[0124] In some embodiments, the bioactive molecule is the therapeutic agent. As used herein, the term “therapeutic agent” refers to a compound used to diagnose, treat, or prevent a disease or disorder in question. Any therapeutic agent known to be beneficial for the diagnosis, treatment, or prevention of a disease or disorder may be included in the pharmaceutical compositions provided herein. Therapeutic agents include pharmacologically active compounds (e.g., antibiotics and anti-inflammatory agents), hormones, DNA (e.g., plasmid DNA), RNA, siRNA, shRNA, antisense RNA, proteins (e.g., antibodies and their fragments, as well as enzymes), lipids, pro-inflammatory molecules, and combinations thereof. Any therapeutic agent may be combined to the extent that such combinations are biocompatible. The amount of therapeutic agent in a pharmaceutical composition will depend on various factors, including, for example, the effective dose of the therapeutic agent required for a particular process of action and the period required for the release of the therapeutic agent. Additional exemplary therapeutic agents and appropriate dosages and regimens are described, for example, in Harrison's Principles of Internal Medicine, 19th edition, edited by L.D. Kasper et al., McGraw-Hill, New York, NY (2015); Physician's Desk Reference, 71st edition, Montvale, NJ, Physician's Desk Reference Inc. (2016); and Goodman and Gilman's: The Pharmacological Basis of Therapeutics, 13th edition, edited by L.L. Brunton et al., McGraw-Hill, New York, NY (2018); each of these is incorporated herein by reference.

[0125] Use of self-assembling peptides and pharmaceutical compositions The self-assembling peptides and pharmaceutical compositions described herein may be used in a variety of in vitro and in vivo applications, including cell culture and clinical applications.

[0126] cell culture In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used in the culture of cells (e.g., isolated human cells). For example, a hydrogel containing the self-assembling peptides described herein may serve as a substrate for cell culture. As described above, the hydrogel formed by the self-assembling peptides described herein may contain one structure or higher-order structures (e.g., nanofibers and three-dimensional meshes) that provide a spatiotemporal substrate for cells. Advantageously, since the hydrogel can be formulated at a physiological pH, it may provide a suitable environment for cell growth and proliferation.

[0127] Any of the cell types described in detail above may be cultured in the hydrogels described herein. Cells may be seeded directly onto pre-formed hydrogels, or they may be mixed with aqueous pharmaceutical compositions that, upon gelation, form a hydrogel. The liquid phase of the hydrogel may be supplemented with one or more culture medium components (e.g., growth factors and serum) to provide factors necessary to promote cell survival and proliferation. Appropriate conditions and factors necessary to maintain cell survival and growth are known in the art (see, for example, Freshney, Culture of Animal Cells: A Manual of Basic Technique, Wiley-Liss, New York, NY (2000); and Cells: A Laboratory Manual (edited by Spector, DL, Goldman, RD, and Leinwand, LA), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1998)).

[0128] The self-assembling peptides and pharmaceutical compositions provided herein may be used in a variety of applications, including, for example, cell culture, cosmetics (e.g., skin and hair care products), tissue engineering, coatings (e.g., for medical devices such as contact lenses), lubricants (e.g., eye drops or joint lubricants), hemostatic agents, desiccants, bone grafts, artificial vitreous humor, artificial lenses, drug delivery devices, water-retaining materials, and biomaterial applications.

[0129] Clinical use In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used in a variety of clinical applications. Accordingly, various methods of treating a subject by administering an effective amount of the pharmaceutical compositions described herein to a specific site in or on the subject are provided herein. As described above, a particular advantage of the pharmaceutical compositions described herein is that they can be formulated at physiological pH (e.g., about pH 6 to about pH 8), and therefore, compared to pharmaceutical compositions of self-assembling peptides formulated at acidic or basic pH, they may result in reduced pain and reduced tissue damage upon contact with the subject.

[0130] The amount, e.g., volume or concentration, of a pharmaceutical composition administered to a subject (e.g., by application or injection) may vary depending on the form of the pharmaceutical composition (e.g., as an aqueous solution or hydrogel) and the route of administration used. The exact formulation, route of administration, volume, and amount of the pharmaceutical composition can be selected considering the subject's symptoms and the specific target area or location to which the pharmaceutical composition is administered. Specific dosages and treatment regimens for a particular subject may depend on a variety of factors, including the specific self-assembling peptide, the dimensions of the area being treated, the desired thickness of the resulting hydrogel (if an aqueous solution is administered), and the length of treatment time. Other factors that may influence specific dosages and treatment regimens include age, weight, health status, sex, administration time, degradation rate, disease, severity, and course of symptoms or signs. In some embodiments, the pharmaceutical composition may be administered in a single dose. In other embodiments, the pharmaceutical composition may be administered in more than one dose (e.g., two, three, four, five, six, or more doses).

[0131] In some embodiments, a pharmaceutical composition in the form of a prepolymerized hydrogel is administered to the subject. In some embodiments, the pharmaceutical composition is a hydrogel formed in vitro and administered to a desired location on the subject.

[0132] In some embodiments, a pharmaceutical composition in the form of an aqueous solution is administered to the subject. A hydrogel may be formed in vivo after administration of the aqueous solution. Since the self-assembling peptide polymerizes in response to changes in tonicity, a hydrogel may be formed upon contact with the subject during or immediately after administration.

[0133] The pharmaceutical composition can be administered by introducing a delivery device into or near a predetermined or desired target area of the subject. The delivery device can be a conventional device or can be designed to achieve at least one of reaching a specific target area, achieving a specific dosing regimen, delivering a specific target volume, amount or concentration, and delivering precisely to the target area. Suitable delivery devices include, but are not limited to, syringes, pipettes, tubes, catheters, syringe catheters, infusion pumps, and other needle-based devices to a predetermined or desired target area of the subject. The gauge of the injection needle can be selected to provide an appropriate flow of the pharmaceutical composition from the syringe to the target area.

[0134] The effective amount can include a pharmaceutical composition in a volume of from about 0.1 milliliter (mL) to about 100 mL (when administered as an aqueous solution). In some embodiments, the effective amount can be a pharmaceutical composition of from about 0.1 mL to about 10 mL. In some embodiments, the effective amount can be a pharmaceutical composition of from about 0.5 mL to about 5 mL. In some embodiments, the effective amount can be a pharmaceutical composition of from about 1 mL to about 5 mL. In some embodiments, the effective amount can be a pharmaceutical composition of about 0.5 mL. In some embodiments, the effective amount can be a pharmaceutical composition of about 1.0 mL. In some embodiments, the effective amount can be a pharmaceutical composition of about 1.5 mL. In some embodiments, the effective amount can be a pharmaceutical composition of about 2.0 mL. In some embodiments, the effective amount can be a pharmaceutical composition of about 2.5 mL. In some embodiments, the effective amount can be about 0.1 mL to about 5 mL per 1 cm of the target area. 2 In some embodiments, the effective amount can be about 0.1 mL to about 5 mL per 1 cm of the target area. 2 In some embodiments, the effective amount can be about 1 mL per 1 cm of the target area. 2 In some embodiments, the effective amount can be about 2 mL per 1 cm of the target area.

[0135] The pharmaceutical composition can be administered to the subject by any suitable route known in the art including, but not limited to, injection, implantation, microinjection, and direct application. Administration via injection includes, but is not limited to, intradermal, intramuscular, intracutaneous, subdermal, and subcutaneous injection.

[0136] In some embodiments, the pharmaceutical composition is administered to a subject in a single dose at one site to produce the desired result (e.g., by injection or implantation). In some embodiments, the pharmaceutical composition is administered as several doses to produce the desired result. In some embodiments, if multiple doses of the pharmaceutical composition are administered, each dose is given after a specific period of time, for example, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 1 month, 2 months, 3 months or more.

[0137] In some embodiments, a hydrogel formed or present at a site of interest may be removed by contacting the hydrogel with a solution containing an isotonic agent at a hypotonic concentration compared to the hydrogel. In some embodiments, a hydrogel formed or present at a site of interest may be removed by contacting the hydrogel with water. In some embodiments, a hydrogel formed or present at a site of interest may be removed by contacting the hydrogel with a hypotonic solution relative to the site of interest. In some embodiments, a hydrogel formed or present at a site of interest may be removed by contacting the hydrogel with an isotonic solution containing sugars (e.g., dextrose and sucrose) relative to the site of interest. In some embodiments, a hydrogel formed or present at a site of interest may be removed by contacting the hydrogel with an isotonic solution (e.g., NaCl-"physiological saline") containing a salt having a lower ionic strength compared to salts in body fluids.

[0138] The hydrogel may be exposed to the solution for a sufficient amount of time to break down the hydrogel (e.g., break down its structure or form) and / or to detach the hydrogel from the site of interest. In some embodiments, the hydrogel may be exposed to the solution for about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 1 hour, about 2 hours, about 3 hours, or longer.

[0139] In some embodiments, the pharmaceutical compositions described herein may be used as delivery devices that specifically target the release of bioactive molecules or therapeutic agents. In some embodiments, the pharmaceutical compositions may be formulated to allow the spontaneous release of bioactive molecules or therapeutic agents (e.g., after administration to a subject). In some embodiments, the pharmaceutical compositions may be formulated to allow the controlled release of bioactive molecules or therapeutic agents. In some embodiments, the bioactive molecules or therapeutic agents are released from the pharmaceutical composition (e.g., a hydrogel) over a long period of time (e.g., about 12 hours to about 2 months). In some embodiments, the bioactive molecules or therapeutic agents are released from the pharmaceutical composition over about 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, or longer. The release kinetics will vary depending on the amino acid sequence of the self-assembling peptide present in the pharmaceutical composition, the concentration of the self-assembling peptide in the pharmaceutical composition, and the biochemical and physical properties of the bioactive molecules or therapeutic agents.

[0140] 1. Gastrointestinal tract obstruction The self-assembling peptides and pharmaceutical compositions described herein may be used to prevent gastrointestinal obstruction of a target area. Methods for preventing gastrointestinal obstruction using pharmaceutical compositions containing self-assembling peptides are known in the art and can be adapted to deliver the pharmaceutical compositions and self-assembling peptides described herein (see, for example, U.S. Patent No. 9,724,448, whose entire contents are expressly incorporated herein by reference). For example, endoscopic mucosal resection (EMR) and endoscopic submucosal dissection (ESD) are primary surgical options for removing lesions such as polyps, ulcers, and cancerous tumors of the gastrointestinal tract (see, for example, Wallace (2017) Gastroenterol.Hepatol. (NY) 13(6):371-4). Although EMR and ESD are minimally invasive procedures, gastrointestinal obstruction can be caused by scar contraction / atrophy during the healing process. One form of gastrointestinal obstruction may be stricture, narrowing of a tubular organ or structure such as the gastrointestinal tract, which can lead to partial or complete obstruction of the gastrointestinal tract. Methods are provided for preventing stricture of the gastrointestinal tract of interest (e.g., one or more of the mouth, throat, esophagus, stomach, small intestine, large intestine, colon, or rectum). In some embodiments, an effective amount of the pharmaceutical composition described herein (e.g., an aqueous solution) is administered to a site in the gastrointestinal tract of interest using a medical device (e.g., a syringe, pipette, tube, syringe catheter, catheter, or endoscope). In some embodiments, the aqueous solution forms a hydrogel at the administration site, thereby enabling prevention of stricture at the administration site. In some embodiments, self-assembling peptides may promote mucosal epithelial formation to prevent or reduce postoperative scar formation, which may contribute to the prevention or reduction of gastrointestinal obstruction or stricture. In some embodiments, the hydrogel provides a scaffold for cell infiltration that promotes healing at the administration site of the hydrogel.

[0141] In some embodiments, a pharmaceutical composition provided herein (e.g., an aqueous solution) may be administered (e.g., by injection) into the submucosa of a site in the gastrointestinal tract of interest before performing excision of the lesion (e.g., using EMR or ESD). The submucosa is a thin layer of connective tissue with a loose structure. In some embodiments, injection of the pharmaceutical composition into the submucosa causes blistering and lifts the lesion upward, thereby facilitating its removal. In some embodiments, the pharmaceutical composition causes the submucosa to rise for a period of about 20 minutes to about 1 hour. In some embodiments, the pharmaceutical composition causes the submucosa to rise for about 20 minutes, about 30 minutes, about 40 minutes, about 50 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, or longer.

[0142] 2. Tissue repair or regeneration The self-assembling peptides and pharmaceutical compositions described herein may be used to promote or enhance the repair or regeneration of a tissue of interest. These self-assembling peptides and pharmaceutical compositions may promote or enhance distinct types of tissue, including, but not limited to, skin, bone, cartilage, nerve tissue, ligaments, tendons, vascular tissue, and muscle (e.g., cardiac tissue). In some embodiments, the subject has a congenital disorder or condition that necessitates tissue repair or regeneration. In some embodiments, the subject has experienced injury that necessitates tissue regeneration. Injury may result from surgery, trauma, stroke, tumor, disease, or disorder (e.g., neurodegenerative disease or disorder). The methods and compositions described herein may restore the structural and / or functional integrity of a tissue (e.g., to its pre-injury structural and / or functional state). Methods for promoting tissue regeneration using hydrogels containing self-assembling peptides are known in the Art and can be adapted for use with the self-assembling peptides and hydrogels described herein (see, for example, U.S. Patent No. 7,846,891, U.S. Patent Application Publication Nos. 2016 / 0362451 and U.S. Patent Application Publication Nos. 2017 / 0128172, whose entire contents are expressly incorporated herein by reference).

[0143] In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to improve or treat the effects of tissue degeneration in organs, to repair damage to organs or other body structures, or to form organs or other body structures. Such organs or body structures include, but are not limited to, vascular tissue, brain, nerve tissue, esophagus, fallopian tubes, heart, intestines, gallbladder, kidneys, liver, lungs, ovaries, pancreas, prostate, bladder, bone, spinal cord, spleen, stomach, testes, thymus, thyroid, trachea, ureters, urethra, uterus, and skin.

[0144] Tissue repair and regeneration can be enhanced by supplying self-assembling peptides and pharmaceutical compositions with bioactive molecules such as growth factors, cell adhesion molecules, and integrins. In some embodiments, the bioactive molecules are present in the pharmaceutical composition. In some embodiments, cells producing one or more bioactive molecules are present in the pharmaceutical composition. For example, genetically modified cells producing and / or secreting one or more bioactive molecules may be present in the pharmaceutical composition.

[0145] The self-assembling peptides and pharmaceutical compositions described herein may be used to promote the regeneration or repair of ocular tissues (e.g., the optic nerve, corneal stroma, and lens cortex). For example, the self-assembling peptides and pharmaceutical compositions described herein may be used to treat subjects with retinopathy or retinal / macular disorders.

[0146] In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to promote or enhance the repair or regeneration of nerve tissue at a target injury site. For example, when administered to an injury site, the pharmaceutical compositions described herein provide an environment that allows for the repair or regeneration of nerve tissue and axonal extension at the injury site.

[0147] 3. Wound healing and antibacterial bandages The self-assembling peptides and pharmaceutical compositions described herein may be used to promote wound healing or skin reconstruction, or to treat wounds (e.g., burns), in subjects requiring promotion or treatment. For example, the pharmaceutical compositions described herein may be applied directly to a wound, or may be adapted for use in gauze or sheets to promote wound healing, promote skin reconstruction, or treat a wound. For example, in some embodiments, the pharmaceutical compositions described herein may be injected into a biopsy site or wound site created by a surgical intervention (e.g., tumor removal). The pharmaceutical compositions described herein may also be used to promote the healing of skin lesions and chronic wounds such as diabetic ulcers. Methods for promoting wound healing or skin reconstruction, and methods for treating wounds using self-assembling peptides are known in the Art and may be adapted for use in the self-assembling peptides and pharmaceutical compositions described herein (see, for example, U.S. Patent Application Publication No. 2011 / 0002880 and International Publication No. 2017 / 210416, whose entire contents are expressly incorporated herein by reference).

[0148] In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used as antimicrobial agents. In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used as antimicrobial bandages.

[0149] 4. Hemostasis The self-assembling peptides and pharmaceutical compositions described herein may be used to promote hemostasis in a subject. The self-assembling peptides and pharmaceutical compositions described herein may be used to stop or control blood loss from blood vessels (e.g., arteries, veins, aorta) and organs of a subject's body (e.g., during surgery or after traumatic injury). Methods for promoting hemostasis in a subject are known in the art and may be adapted for use with the self-assembling peptides and pharmaceutical compositions described herein (see, for example, International Publication No. 2017 / 210421, whose entire contents are expressly incorporated herein by reference).

[0150] In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to create a physical barrier to stop or prevent bleeding during surgical or endoscopic procedures (e.g., during hepatectomy, splenectomy, vaginoplasty, cholecystectomy, coronary artery bypass, or femoral bypass). In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to stop or prevent exudative bleeding from blood vessels and solid organ parenchyma. In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to stop or prevent exudative bleeding from vascular anastomoses (e.g., anastomoses to natural or artificial blood vessels). In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to stop or prevent exudative bleeding from small vessels or capillaries of the gastrointestinal tract under consideration (e.g., during endoscopic submucosal dissection (EMD), laparoscopic resection of the gastrointestinal tract, or endoscopic mucosal resection (EMR) of the gastrointestinal tract).

[0151] 5. Local drug / therapeutic agent delivery In one embodiment, the Disclosure provides a combination composition comprising (i) a self-assembling peptide comprising a self-assembling peptide or pharmaceutical composition described herein and (ii) one or more payload agents (e.g., therapeutic agents), wherein the combination composition has a storage modulus of about 0.1 to about 100 Pa (e.g., at a frequency of 5 radians / second and a vibrational stress of 0.1 Pa) and / or a viscosity in the range of about 0.5 Pa to about 50,000 Pa at room temperature. In some embodiments, one or more payload agents (e.g., therapeutic agents) of the combination composition are substantially uniformly distributed within the combination composition.

[0152] In another embodiment, the self-assembling peptides and pharmaceutical compositions described herein may be used for the local delivery of drugs and / or therapeutic agents to affected and / or defective sites in the body. The drug / therapeutic agent may be mixed with the peptide solution and injected using a needle, nozzle, or catheter. The drug can then be delivered and localized to the tissue, and the hydrogel can be slowly released before it is absorbed into the body.

[0153] As used herein, the term “drug” refers to any chemical class of compound or entity, including, for example, polypeptides, nucleic acids, sugars, lipids, small molecules, metals, or combinations thereof. In some embodiments, the drug is a natural product, or includes a natural product, in that it is found in nature and / or obtained from nature. In some embodiments, the drug is one or more entities, or includes such entities, that are designed, manipulated and / or manufactured by human action and / or are not found in nature. In some embodiments, the drug may be available in an isolated or pure form; in some embodiments, the drug may be available in a crude form. In some embodiments, the potential drug is provided as a collection or library, which can be screened, for example, to identify or characterize the activator in it. Some specific embodiments of drugs that may be available according to the present invention include small molecules, antibodies, antibody fragments, aptamers, nucleic acids (e.g., siRNA, shRNA, DNA / RNA hybrids, antisense oligonucleotides, mRNA, CRISPR systems, and ribozymes), peptides, peptide mimes, etc. In some embodiments, the drug is a polymer, or includes a polymer. In some embodiments, the drug is not a polymer and / or substantially does not contain any polymer. In some embodiments, the drug contains at least one polymer portion. In some embodiments, the drug lacks or substantially does not contain any polymer portion. In some embodiments, the drug is cells and / or tissue. In some embodiments, the drug is a cell lysate or contains a cell lysate. In some embodiments, the drug is a cellular material and / or multicellular material (e.g., a microcolumn graft and / or micrograft), or contains such material.

[0154] As used herein, the phrase “therapeutic agent” generally refers to any agent that, when administered to an organism, induces a desired pharmacological effect. In some embodiments, an agent is considered a therapeutic agent if it demonstrates a statistically significant effect across a suitable population. In some embodiments, a suitable population may be a population of model organisms. In some embodiments, a suitable population may be defined by various criteria such as a specific age group, sex, genetic background, pre-existing clinical symptoms, etc. In some embodiments, a therapeutic agent is a substance that can be used to alleviate, improve, relieve, inhibit, prevent, delay the onset of, reduce the severity of, and / or decrease the incidence of one or more symptoms or characteristics of a disease, disorder and / or symptom. In some embodiments, a “therapeutic agent” is an agent that has been or needs to be approved by a government agency before it can be marketed for administration to humans. In some embodiments, a “therapeutic agent” is an agent that requires a prescription for administration to humans.

[0155] In some specific embodiments, the combination composition of the present invention comprises one or more payload agents, e.g., therapeutic agents or detection agents. Such agents include compounds or entities of any chemical class, e.g., polypeptides, nucleic acids, sugars, lipids, small molecules, metals, or combinations thereof. As is evident from the context, in some embodiments, the agent may be, or may include, a cell or organism, or a fraction, extract or component thereof. In some embodiments, the agent is a natural product, or includes a natural product, in that it is found in nature and / or obtained from nature. In some embodiments, the agent is one or more entities, or includes, that are artificial, in that they are designed, manipulated and / or manufactured by human action and / or not found in nature. In some embodiments, the agent may be available in an isolated or pure form; in some embodiments, the agent may be available in a crude form. In some embodiments, the potential agent is provided as a collection or library, for example, that can be screened to identify or characterize the active ingredient in it. Some specific embodiments of the agents that may be used in accordance with the present invention include small molecules (e.g., antibiotics, anticancer agents, anti-analgesics, anti-inflammatory drugs, steroids, antipsychotics), antibodies, antibody fragments, aptamers, nucleic acids (e.g., siRNA, shRNA, DNA / RNA hybrids, antisense oligonucleotides, ribozymes), peptides, peptide mimes, proteins, fusion proteins, vaccines, anticoagulants, cytokines, hormones, enzymes, blood factors, extracellular matrix components, etc. In some embodiments, the agent may be selected from the group consisting of micrograft tissues, drugs (e.g., antibiotics), and biology (e.g., growth factors and / or other molecules / proteins). In some embodiments, the agent is a cytokine (e.g., epidermal growth factor, nerve growth factor, transforming growth factors - alpha and beta, platelet-derived growth factor, insulin-like growth factor, vascular endothelial growth factor).

[0156] In some embodiments, the self-assembling peptides or pharmaceutical compositions described herein, combined with one or more payload agents (e.g., therapeutic agents), are used to treat diseases or disorders, such as diseases or disorders known or suspected to be treated by the therapeutic agents described herein (e.g., infections, cancers, cardiovascular diseases, neurological disorders), and / or for wound healing, bone / cartilage repair / regeneration, and soft tissue regeneration. The self-assembling peptides or pharmaceutical compositions described herein, combined with one or more payload agents (e.g., therapeutic agents), can be administered to a subject in a variety of ways, and the administration is not limited to a particular method. In some embodiments, the self-assembling peptides or pharmaceutical compositions described herein, combined with one or more payload agents (e.g., therapeutic agents), are administered to or applied to a subject by a device, medical device, implant, dental implant, breast implant, prosthesis, needle, stent, or catheter. Additional methods of administration are described, for example, in U.S. Patent Application Publication No. 2011-0002880; International Publication No. 2008 / 073395; U.S. Patent Application Publication No. 2011 / 0201541; U.S. Patent Application Publication No. 2014-0329914; U.S. Patent Application Publication No. 2015-0105336; International Publication No. 2014 / 136081; International Publication No. 2014 / 141143; and U.S. Patent No. 7846891, each of which is incorporated herein by reference in whole for any and all purposes.

[0157] Additional methods and embodiments can be found in International Publication No. 2017 / 120092, which is incorporated herein by reference in its entirety for all purposes.

[0158] 6. Air Leakage Blockage In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used for air leak obstruction. In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to treat alveolar cysts. Embodiments provide a method for treating a target alveolar cyst and / or air leak. This method may include the step of introducing a delivery device into a target region of the target alveolar cyst. This method may also include the step of positioning the end of the delivery device in the target region where treatment of the alveolar cyst is desired. This method may also include the step of administering a solution containing an effective amount and effective concentration of the self-assembling peptide or pharmaceutical composition described herein to the target region through the delivery device to treat the alveolar cyst by forming a barrier under physiological conditions of the target region. This method may also include the step of removing the delivery device from the target region.

[0159] Additional methods and embodiments can be found in International Publication No. 2013 / 030673, International Publication No. 2015 / 138473, and U.S. Patent No. 10245299, each of which is incorporated herein by reference in whole for any and all purposes.

[0160] 7. Prevention and reduction of adhesion In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to prevent or reduce adhesion to biological tissues. In various embodiments, the present invention provides a method for reducing adhesion to biological tissues, comprising the step of administering an effective amount of the self-assembling peptides and pharmaceutical compositions described herein to a biological tissue, wherein the self-assembling peptides and pharmaceutical compositions described herein reduce adhesion to biological tissues.

[0161] In various embodiments, the present invention provides a method for promoting anti-adhesion, comprising the steps of: introducing a delivery device into a target region; positioning the end of the delivery device in a target region where anti-adhesion is desired; administering a solution containing an effective amount and effective concentration of a self-assembling peptide or pharmaceutical composition described herein to the target region through the delivery device to promote anti-adhesion; and removing the delivery device from the target region.

[0162] In some embodiments, the biological tissue includes the epicardium, intraperitoneal cavity, cecum, intestine, preferably the large intestine, and / or colon. In some embodiments, the target region includes the epicardium, intraperitoneal cavity, cecum, intestine, preferably the large intestine, and / or colon.

[0163] Additional methods and embodiments can be found in U.S. Patent Application Publication No. 2019-0091376, which is incorporated herein by reference in its entirety for all purposes.

[0164] 8. In vitro cell culture In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used in 2D cell culture and / or 3D cell culture.

[0165] 9. Interosseous space filler In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to fill interosseous spaces.

[0166] In some embodiments, a method for filling a bone gap in a target is provided. In some embodiments, the method may involve the steps of introducing a delivery device into a bone in a target; positioning the end of the delivery device proximal to a bone gap in which bone growth promotion is desired; administering a solution containing an effective amount and concentration of the self-assembling peptide or pharmaceutical composition of the present disclosure, ranging from about 0.1 w / v percent to about 5 w / v percent of the peptide, through the delivery device to promote bone growth at the target site, thereby forming a hydrogel scaffold under physiological conditions; and removing the delivery device from the target.

[0167] In one or more embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used as interosseous space fillers (BVFs) that, after administration at a target site, are reabsorbed during the healing process and replaced by bone. Peptide hydrogels may be placed in interosseous spaces or gaps in the skeletal system. In certain embodiments, self-assembling peptides and their self-assembling structures may be used as cell culture supports for the repair and replacement of various tissues, as well as as scaffolds for encapsulating living cells. The self-assembling peptides and pharmaceutical compositions described herein may promote tissue regeneration and the production of associated extracellular matrix proteins. In at least some embodiments, the self-assembling peptides and pharmaceutical compositions described herein are non-immunogenic and represent an improvement over existing materials for this indication, including demineralized freeze-dried bone (DFDBA) preparations.

[0168] Additional methods and embodiments can be found in U.S. Patent Application Publication No. 2017-0128622, which is incorporated herein by reference in its entirety for all purposes.

[0169] 10. Artificial tears for dry eyes In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to treat dry eye. In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used in artificial tears.

[0170] 11. Articular cartilage repair In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to repair articular cartilage. In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used as injectable compositions. Not bound by theory, injectable hydrogels comprising the self-assembling peptides and pharmaceutical compositions described herein may enhance their therapeutic efficacy and improve their ease of administration. An ideal injectable scaffold for cartilage regeneration should typically meet the following criteria: (i) ease of administration under physiological conditions, (ii) guaranteed injectability (gelation upon injection via chemical or physical crosslinking), (iii) excellent biocompatibility and potential biodegradability, (iv) ability to mimic cartilage ECM function and promote cellular chondrolytic capacity, (v) ability to readily fill defective sites within the joint and integrate with the surrounding natural cartilage tissue rather than easily migrating, and (vi) a sustained-release profile, where relevant to topical drug delivery.

[0171] 12. Cosmetic applications (dermal fillers in cosmetic surgery) In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used as skin fillers. In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used as injectable compositions. Although not bound by theory, injectable hydrogels comprising the self-assembling peptides and pharmaceutical compositions described herein may enhance their therapeutic efficacy and improve their ease of administration. An ideal injectable scaffold for cartilage regeneration should typically meet the following criteria: (i) ease of administration under physiological conditions, (ii) guaranteed injectability (gelation upon injection via chemical or physical crosslinking), (iii) excellent biocompatibility and potential biodegradability, (iv) ability to mimic cartilage ECM function and promote cellular chondrolytic capacity, (v) ability to readily fill defect sites within joints and integrate with surrounding natural cartilage tissue rather than easily migrating, and (vi) a sustained-release profile, where relevant to topical drug delivery.

[0172] In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to plump the lips. In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to fill in or smooth wrinkles. In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to soften facial wrinkles. In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to enhance shallow contours. In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to reconstruct facial contour deformities. In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to improve the appearance of sunken scars. In some embodiments, the self-assembling peptides and pharmaceutical compositions described herein may be used to reduce or eliminate shadows under the lower eyelids.

[0173] kit In another aspect, the Disclosure provides kits and manufactured articles comprising pharmaceutical compositions and / or self-assembling peptides as described herein. The kits may include instructions for the proper preparation and / or use of the components therein. The kits may further include useful tools for preparing and / or delivering the pharmaceutical compositions or self-assembling peptides to subjects (e.g., human subjects) as described herein.

[0174] In some embodiments, the kit includes separate containers, dividers, or compartments for the pharmaceutical composition and informational materials. For example, the pharmaceutical composition may be contained in a bottle, vial, or syringe, and the informational materials may be contained in a plastic sleeve or packet. In some embodiments, the separate elements of the kit are contained within a single, undivided container. For example, the pharmaceutical composition or self-assembling peptide may be contained in a bottle, vial, or syringe with informational materials attached in the form of a label.

[0175] In some embodiments, the kit comprises a plurality of individual containers, e.g., packs of individual containers, each containing one or more unit dosage forms of the pharmaceutical composition or self-assembling peptide. For example, the kit may comprise a plurality of syringes, ampoules, foil packets, or blister packs, each containing a single unit dose of the pharmaceutical composition or self-assembling peptide. In some embodiments, the components of the kit are stored in sealed vials equipped with, for example, rubber or silicone closures (e.g., polybutadiene or polyisoprene closures). In some embodiments, the components of the kit are stored under inert conditions (e.g., under another inert gas such as nitrogen or argon). In some embodiments, the components of the kit are stored under anhydrous conditions (e.g., with a desiccant). In some embodiments, the components of the kit are stored in light-shielding containers, such as amber vials. In some embodiments, the containers of the kit may be airtight and / or waterproof.

[0176] The kit may further include a solution containing an isotonic agent. These solutions may be packaged separately from or in combination with the pharmaceutical composition containing the self-assembling peptide provided herein.

[0177] For example, manufactured articles including syringes, automatic infusion devices, tubing, and catheters (with or without guidewires) are also provided. In some embodiments, manufactured articles may be pre-filled with the self-assembling peptides or pharmaceutical compositions described herein. These manufactured articles may be separate from or included in the kits described herein.

[0178] Further aspects and embodiments of the present invention are shown in the following numbered paragraphs: 1. The amino acid sequence shown below: [(X)i(Y)j(Z)k(Y)l]m(X)n Formula I, [(Y)i(X)j(Y)k(Z)l]m(Y)n Formula II, [(Z)i(Y)j(X)k(Y)l]m(Z)n Equation III, or [(Y)i(Z)j(Y)k(X)l]m(Y)n Equation IV (In the formula, each (X) is independently an ionic polar amino acid, each (Y) is independently a hydrophobic amino acid, each (Z) is independently a nonionic polar amino acid, each i, j, k, and l is independently an integer ≥ 1, m is an integer ≥ 2, and n = 0 or an integer ≥ 1) A pharmaceutical composition containing a self-assembling peptide.

[0179] 2. The pharmaceutical composition according to paragraph 1, wherein the self-assembling peptide comprises the amino acid sequence shown in formula I.

[0180] 3. The pharmaceutical composition according to paragraph 1, wherein the self-assembling peptide comprises the amino acid sequence shown in formula II.

[0181] 4. The pharmaceutical composition according to paragraph 1, wherein the self-assembling peptide comprises the amino acid sequence shown in formula III.

[0182] 5. The pharmaceutical composition according to paragraph 1, wherein the self-assembling peptide comprises the amino acid sequence shown in formula IV.

[0183] 6. A pharmaceutical composition according to any one of paragraphs 1 to 5, wherein each (X) is a basic amino acid.

[0184] 7. The pharmaceutical composition according to paragraph 6, wherein the basic amino acid is selected from the group consisting of arginine, lysine, histidine, and ornithine.

[0185] 8. A peptide described in any one of paragraphs 1 to 5, wherein each (X) is an acidic amino acid.

[0186] 9. The peptide according to paragraph 8, wherein the acidic amino acid is selected from the group consisting of aspartic acid and glutamic acid.

[0187] 10. A pharmaceutical composition according to any one of paragraphs 1 to 9, wherein each (Y) is selected from the group consisting of alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, and glycine.

[0188] 11. A pharmaceutical composition according to any one of paragraphs 1 to 10, wherein each (Z) is selected from the group consisting of serine, threonine, tyrosine, cysteine, glutamine, asparagine, and methionine.

[0189] 12. A pharmaceutical composition according to any one of paragraphs 1 to 11, wherein at least one of i, j, k, and l is independently an integer 1.

[0190] 13. A pharmaceutical composition according to any one of paragraphs 1 to 12, wherein at least one of i, j, k, and l is independently an integer 2.

[0191] 14. A pharmaceutical composition according to any one of paragraphs 1 to 13, wherein m is independently an integer of 2 or 3.

[0192] 15. The pharmaceutical composition according to any one of paragraphs 1 to 14, wherein the self-assembling peptide comprises the amino acid sequence shown in SEQ ID NOs: 1 to 20.

[0193] 16. The pharmaceutical composition according to any one of paragraphs 1 to 15, wherein the self-assembling peptide comprises an N-terminal functional group, a C-terminal functional group, or both.

[0194] 17. The pharmaceutical composition according to paragraph 16, wherein the N-terminal functional group is selected from the group consisting of acetyl, formyl, pyroglutamyl (pGlu), biotin, polyethylene glycol (PEG), urea, alkylamine, carbamate, sulfonamide, dansyl, 2,4-dinitrophenyl, fluorescein, 7-methoxycoumarin acetate, 9-fluorenylmethyloxycarbonyl, palmitic acid, succinyl, chloroacetyl, maleimide, benzyloxycarbonyl, bromoacetyl, nitrilotriacetyl, tert-butoxycarbonyl, 4-hydroxyphenylpropionic acid, allyloxycarbonyl, butyric acid, fatty acid, and trityl.

[0195] 18. The pharmaceutical composition according to paragraph 16, wherein the C-terminal functional group is selected from the group consisting of amide, N-alkylamide, aldehyde, ester, alcohol, para-nitroanilide (pNA), 7-amino-4-methylcoumarin (Amc), hydrazide, hydroxamic acid, chloromethyl ketone, p-nitroaniline, para-nitrophenol, hydroxysuccinimide ester, fluoromethyl ketone, cysteamide, 9-fluorenemethyl (Fm) ester, allyl ester, 2,4-dimethoxybenzyl ester, 2-phenylisopropyl ester, p-nitrobenzyl ester, and 2-chlorotrityl ester.

[0196] 19. The pharmaceutical composition according to any one of paragraphs 1 to 16, wherein the self-assembling peptide comprises the amino acid sequence shown in SEQ ID NOs. 21 to 40.

[0197] 20. The pharmaceutical composition according to any one of paragraphs 1 to 19, wherein the self-assembling peptide further comprises at least one biologically active peptide motif.

[0198] 21. A pharmaceutical composition according to any one of paragraphs 1 to 20, wherein at least one biologically active peptide motif is located at the N-terminus of a self-assembling peptide.

[0199] 22. The pharmaceutical composition according to any one of paragraphs 1 to 20, wherein at least one biologically active peptide motif is present at the C-terminus of the self-assembling peptide.

[0200] 23. The pharmaceutical composition according to any one of paragraphs 20 to 22, wherein 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.

[0201] 24. The pharmaceutical composition according to any one of paragraphs 20 to 22, wherein at least one biologically active peptide motif comprises an amino acid sequence shown in any one of SEQ ID NOs: 41 to 70.

[0202] 25. The pharmaceutical composition according to any one of paragraphs 1 to 20, wherein the self-assembling peptide comprises an amino acid sequence shown in SEQ ID NOs: 71 to 90.

[0203] 26. The pharmaceutical composition according to any one of paragraphs 1 to 25, further comprising an isotonic agent.

[0204] 27. The pharmaceutical composition according to paragraph 26, wherein the isotonic agent is present at a concentration of about 0.01 M to about 0.3 M.

[0205] 28. The pharmaceutical composition according to paragraph 26, wherein the isotonic agent is present at a concentration of about 0.15 M.

[0206] 29. The pharmaceutical composition according to any one of paragraphs 1 to 28, having a pH of about 6 to about 8.

[0207] 30. The pharmaceutical composition according to any one of paragraphs 1 to 28, having a pH of about 7 to about 7.5.

[0208] 31. The pharmaceutical composition according to paragraph 29 or paragraph 30, wherein the net charge of the self-assembling peptide is greater than +1 or equal to +1 or less than -1 or equal to -1.

[0209] 32. The pharmaceutical composition according to paragraph 29 or paragraph 30, wherein the net charge of the self-assembling peptide is from about +1 to about +6.

[0210] 33. The pharmaceutical composition according to paragraph 29 or paragraph 30, wherein the net charge of the self-assembling peptide is from about -1 to about -6.

[0211] 34. The pharmaceutical composition according to any one of paragraphs 1 to 33, wherein the concentration of the self-assembling peptide is from about 0.01% (w / v) to about 10% (w / v).

[0212] 35. The pharmaceutical composition according to any one of paragraphs 1 to 33, wherein the concentration of the self-assembling peptide is from about 0.1% (w / v) to about 5% (w / v).

[0213] 36. The pharmaceutical composition according to any one of paragraphs 1 to 33, wherein the concentration of the self-assembling peptide is from about 0.5% (w / v) to about 1.5% (w / v).

[0214] 37. The pharmaceutical composition according to any one of paragraphs 1 to 33, wherein the concentration of the self-assembling peptide is about 1% (w / v).

[0215] 38. The pharmaceutical composition according to any one of paragraphs 1 to 37, further comprising isolated cells.

[0216] 39. The pharmaceutical composition according to paragraph 38, wherein the isolated cells are mammalian cells.

[0217] 40. The pharmaceutical composition according to paragraph 38 or paragraph 39, wherein the mammalian cells are immune cells, stem cells, chondrocyte progenitor cells, pancreatic progenitor cells, myoblasts, fibroblasts, keratinocytes, nerve cells, glial cells, astrocytes, preadipocytes, adipocytes, vascular endothelial cells, endothelial progenitor cells, mesenchymal cells, neural stem cells, immune cells (e.g., B cells and T cells), smooth muscle progenitor cells, cardiomyocytes, fetal dermal fibroblasts, epidermal keratinocytes, myoblasts, and capillary endothelial cells.

[0218] 41. A pharmaceutical composition according to any one of paragraphs 1 to 40, further comprising a bioactive agent.

[0219] 42. The pharmaceutical composition according to paragraph 41, wherein the bioactive agent is selected from the group consisting of hormones, growth factors, insulin, enzymes, siRNA, shRNA, antisense RNA, antibiotics, antibodies, and anti-inflammatory agents.

[0220] 43. A pharmaceutical composition described in any one of paragraphs 1 to 42, which is an aqueous solution.

[0221] 44. A pharmaceutical composition described in any one of paragraphs 1 to 42, which is a hydrogel.

[0222] 45. The pharmaceutical composition according to paragraph 44, which is a hydrogel having a storage modulus of at least about 10 pascals (Pa).

[0223] 46. ​​A manufactured article containing a pharmaceutical composition described in any one of paragraphs 1 to 45.

[0224] 47. A manufactured article as described in paragraph 46, which is a syringe, vial, automatic infusion device, tube or catheter.

[0225] 48. A method for treating a subject in need of treatment, comprising the step of administering an effective amount of a pharmaceutical composition described in any one of paragraphs 1 to 45 to the subject.

[0226] 49. A method for promoting tissue repair or regeneration in a subject that requires promoting tissue repair or regeneration, the method comprising contacting the tissue of the subject with the pharmaceutical composition according to any one of paragraphs 1 to 45, thereby promoting tissue repair or regeneration of the tissue.

[0227] 50. The method according to paragraph 49, wherein the tissue is skin, bone, cartilage, nerve tissue, ligament, tendon, vascular tissue or muscle.

[0228] 51. The method according to paragraph 49, wherein the tissue is eye tissue.

[0229] 52. The method according to paragraph 49, wherein the tissue is heart tissue.

[0230] 53. The method according to any one of paragraphs 49 to 52, wherein the subject has a congenital disease or disorder that causes a need for tissue repair or regeneration.

[0231] 54. The method according to any one of paragraphs 49 to 52, wherein the subject is suffering from an injury that causes a need for tissue repair or regeneration.

[0232] 55. The method according to paragraph 54, wherein the injury is the result of surgery, trauma, stroke, tumor, or disease or disorder.

[0233] 56. A method for promoting wound healing in a subject that requires promoting wound healing, the method comprising contacting the wound of the subject with the pharmaceutical composition according to any one of paragraphs 1 to 45, thereby promoting wound healing.

[0234] 57. The method according to paragraph 56, wherein the wound includes abrasion, burn, crack, contusion, incision, ulcer, laceration, dissection or scratch.

[0235] 58. A method for stopping or preventing bleeding at a site within a subject, comprising the step of bringing the site into contact with a pharmaceutical composition described in any one of paragraphs 1 to 45, wherein the pharmaceutical composition creates a physical barrier, thereby stopping or preventing bleeding at the site within the subject.

[0236] 59. A method for resecting a lesion from the target gastrointestinal tract, a. The step of bringing a pharmaceutical composition described in any one of paragraphs 1 to 45 into contact with the submucosal layer beneath the lesion, thereby lifting the lesion; b. The step of excising the lesion from the aforementioned part of the gastrointestinal tract of the subject. A method that includes this.

[0237] 60. The method according to paragraph 59, wherein the lesion includes a polyp, ulcer, or tumor.

[0238] 61. The method according to paragraph 59 or paragraph 60, wherein the lesion is located in a region of the gastrointestinal tract of the subject, selected from the mouth, throat, esophagus, stomach, small intestine, large intestine, colon, and rectum.

[0239] 62. A method for culturing cells, comprising the step of bringing the cells into contact with a pharmaceutical composition described in any one of paragraphs 1 to 45.

[0240] The following examples are for illustrative purposes only and are not limiting. Many variations of the art will become apparent to those skilled in the art upon consideration of this disclosure. Therefore, the scope of the art should be determined without reference to the examples, but rather by reference to the appended claims, along with the full scope of the equivalents. [Examples]

[0241] material and method The experiments described in the following examples were carried out using the following materials and methods.

[0242] Peptide synthesis All peptides used in the examples described herein were synthesized by conventional solid-state peptide synthesis using an automated peptide synthesizer.

[0243] Viscosity measurement The peptide, formulated as an aqueous solution (200 μL), was placed between rheometer plates (DHR-1, TA Instruments, 20 mm plate with a 500 μm measurement shape), and the viscosity was measured for 0.001 seconds. -1 ~100 seconds -1 The measurement was taken using the shear rate. Thixotropy measurement.

[0244] The peptide, formulated as an aqueous solution (200 μL), is placed between the plates of a rheometer (DHR-1, TA Instruments, 20 mm plate with a 500 μm measurement shape), and the sample is measured for 1000 seconds. -1 The plate was subjected to high-speed plate rotation for 1 minute at a shear rate of , and the storage modulus was measured in 1 radian-second intervals. -1 The measurement was taken for 10 minutes at the given angular frequency.

[0245] Frequency sweep test Peptides formulated as aqueous solutions were placed between rheometer plates (DHR-1, TA Instruments, 20 mm plates with 500 μm or 40 mm cone measurement shapes, and plates with a 2.0° cone angle measurement shape), and frequency stress sweep tests were performed at frequencies from 0.1 Hz to 10 Hz and a strain of 0.1%; measurements were performed after a 2-minute relaxation time at 37°C. To perform frequency sweep tests after exposure to Dulbecco's Modified Eagle Medium (DMEM), 10 mL of DMEM was added to the chamber surrounding the plates, the samples in the plates were immersed in the DMEM, and the frequency test was performed after 20 minutes as described.

[0246] Endoscopic mucosal resection and endoscopic submucosal dissection Endoscopic mucosal and submucosal dissection was performed using an in vivo porcine animal model, as described in Uraoka et al. (2009) Drug Des. Devel. Ther. 2:131-8, incorporated herein by reference. Briefly, 2 mL of a selected peptide formulation was injected between the muscle and submucosa of the porcine stomach, and the submucosa was dissected using an electro-knife. The macroscopic appearance of the injection site and the submucosal tissue dissected at the injection site was analyzed.

[0247] Example 1. Determination of the net charge of exemplary self-assembling peptides in formulations across various pH ranges. Several exemplary self-assembling peptides described herein (Table 5; SEQ ID NOs. 21-40) were designed to have a net positive or negative charge when formulated at pH 7.5. In contrast, the self-assembling peptides have a net zero charge at the same pH. The net charges of the exemplary self-assembling peptides in solution at pH 7.5 were determined and are shown in Table 5. The self-assembling peptides RADA16, IEIK13, and KLD12 (SEQ ID NOs. 91-93, respectively) had a net charge of nearly zero at pH 7.5, while the newly developed self-assembling peptides had a net positive or net negative charge at the same pH.

[0248] As shown in Figures 1 to 3, this net charge behavior spreads around the pH 7.5 test point shown in Table 5. For example, Figure 1 shows the net charge as a function of pH for KLNL12 (sequence number 21, approximately +3 near neutral pH). Figure 2 shows the net charge as a function of pH for NLEL12 (sequence number 33, approximately -3 near neutral pH). Figure 3 shows the net charge as a function of pH for RADA16 (sequence number 91, approximately 0 near neutral pH).

[0249] Table 5. Net charge of exemplary self-assembling peptides at pH 7.5. [Table 5-1] [Table 5-2] * For comparison, an example of a conventional self-assembling peptide.

[0250] Figures 4 and 5 provide molecular models of the atomic structure and net charge of exemplary self-assembling peptides. Figure 4 shows that RANA16 (SEQ ID NO: 39) has a net charge of +4 at pH 7.5. In contrast, Figure 5 shows that RADA16 (SEQ ID NO: 91) has a net charge of 0 at pH 7.5. The structures were computationally simulated using MarvinSketch and both exhibit a β-sheet conformation. Despite RANA16 (SEQ ID NO: 39) and RADA16 (SEQ ID NO: 91) exhibiting similar structural conformations at pH 7.5, the net charges of the two peptides differ significantly, with RANA16 (SEQ ID NO: 39) having a net positive charge of +4, while RADA16 (SEQ ID NO: 91) has a net positive charge of zero.

[0251] Example 2. Characterization of the solubility of exemplary self-assembling peptides in aqueous pharmaceutical compositions formulated at pH 2.5 and 7.5. Several exemplary self-assembling peptides described herein (see Table 5; SEQ ID NOs: 21-40) have a non-zero net charge, high solubility when formulated in a pH 7.5 solution, and are designed to transition from a liquid state to a hydrogel after administration (e.g., after injection into a subject, or after misculation with a solution containing a specific concentration of isotonic agent (e.g., a salt)). In contrast, the self-assembling peptides RADA16, IEIK13, and KLDL12 (SEQ ID NOs: 91-93, respectively) have a zero net charge and must be formulated in an acidic pH solution to retain their ability to conditionally transition from a liquid state to a hydrogel after administration.

[0252] To determine the solubility of exemplary self-assembling peptides in aqueous pharmaceutical compositions formulated at acidic and neutral pH, the appearance of the pharmaceutical compositions was evaluated. A homogeneous and clear appearance reflects sufficient solubility of the peptide in solution and minimal hydrogel formation. Phase separation and a turbid appearance reflect insufficient solubility of the peptide in solution and / or hydrogel formation.

[0253] The appearance of exemplary self-assembling peptides at pH 2.5 and pH 7.5 is shown below in Table 6 and Figure 20. The aqueous pharmaceutical composition at pH 7.5 containing the newly described self-assembling peptide was clear and homogeneous, while the previously disclosed self-assembling peptides RADA16 (SEQ ID NO: 91), IEIK13 (SEQ ID NO: 92), and KLDL12 (SEQ ID NO: 93) at pH 7.5 showed phase separation and a turbid appearance (see Table 6 and Figures 20A-20C).

[0254] Table 6. Appearance of exemplary self-assembling peptides at pH 2.5 and pH 7.5. [Table 6-1] [Table 6-2]

[0255] Example 3. Characterization of the solubility of aqueous pharmaceutical compositions formulated at pH 7.5 and various ionic strengths. The self-assembling peptides described herein (Table 5; SEQ ID NOs: 21-40) have a non-zero net charge and, when formulated as an aqueous solution at pH 7.5 with isotonic ionic strength (e.g., 0.15 M sodium chloride ions), exhibit high solubility and are designed to transition from a liquid state to a hydrogel after administration. To determine the solubility of aqueous pharmaceutical compositions formulated at pH 7.5 in the presence of various ionic strengths, aqueous pharmaceutical compositions with different concentrations of sodium chloride were prepared and their appearance was evaluated. A homogeneous and clear appearance reflects sufficient solubility of the peptide in the solution and minimal hydrogel formation. Phase separation and a turbid appearance reflect insufficient solubility of the peptide in the solution and / or hydrogel formation.

[0256] As shown in Table 7, generally, an increase in the amount of hydrophobic residues correlated with a decrease in solubility at higher ionic strengths. Therefore, the amino acid composition of self-assembling peptides can be manipulated to achieve the desired solubility at the desired ionic strength.

[0257] Table 7. Appearance of exemplary self-assembling peptides at different ionic intensities. [Table 7-1] [Table 7-2] * Since the peptide precipitates at around pH 7.5, the formulation was prepared at around pH 2.2-2.3.

[0258] Example 4. Determination of shear reduction and thixotropic behavior of aqueous pharmaceutical compositions containing exemplary self-assembling peptides formulated at pH 7.5 with varying ionic strengths. The self-assembling peptides disclosed herein (Table 5; SEQ ID NOs: 21-40) have a non-zero net charge, high solubility when formulated in an aqueous solution at pH 7.5 in the presence of isotonic ionic strength (e.g., 0.15 M salt ions), and are designed to transition from a liquid state to a hydrogel after administration to a target. In some clinical applications, administration of pharmaceutical compositions containing self-assembling peptides requires exposure of the composition to pressure and / or shear reduction (e.g., during injection through a needle or transfer via a pump-based system). Ideally, pharmaceutical compositions containing self-assembling peptides described herein exhibit viscosity reduction under pressure or shear reduction forces and return to their baseline viscosity after removal from these forces. To determine the shear reduction properties of pharmaceutical compositions of self-assembling peptides formulated at pH 7.5, formulations with various ionic strengths were prepared using different concentrations of sodium chloride and measured at shear rates that increased viscosity. The thixotropic properties of the formulations, as well as the storage modulus over time at specific frequencies and pressures, were also determined.

[0259] Figures 6–8 show the experimentally determined viscosity of aqueous pharmaceutical compositions containing either 1% (w / v) KLNL12 (SEQ ID NO: 21), 1% (w / v) IQIK13 (SEQ ID NO: 28), or 1% (w / v) NLEL12 (SEQ ID NO: 33) at pH 7.5. Figures 9 and 10 show the experimentally determined viscosity of aqueous pharmaceutical compositions containing either 1% KLNL12 (SEQ ID NO: 21) or 1% NLEL12 (SEQ ID NO: 33) and 0.9% NaCl (to mimic physiological conditions) at pH 7.5. The data demonstrate that the aqueous pharmaceutical compositions exhibit complete shear thinning; that is, viscosity decreases with increasing shear rate. This property can favorably facilitate the application and use of the pharmaceutical composition (e.g., by viscosity reduction during pumping) while maintaining the desired viscosity at the application site (e.g., where no shear force is applied).

[0260] Figures 11 and 12 show the thixotropic properties of pharmaceutical compositions containing 1% KLNL12 (SEQ ID NO: 21) or 1% NLEL12 (SEQ ID NO: 33) and 0.9% NaCl at pH 7.5 after shear stress has been removed. The aqueous pharmaceutical compositions were flowed at a shear rate of 1000 1 / second for 1 minute, and their storage modulus was tested over time at a frequency of 10 radians / second and a stress of 0.1 Pa. The tested aqueous pharmaceutical compositions exhibited thixotropic properties. This property can advantageously facilitate the application and use of pharmaceutical compositions because it indicates that the pharmaceutical compositions exhibit predictable and reliable properties after the application of mechanical stress.

[0261] Example 5. Determination of the rheological properties of a pharmaceutical composition containing a self-assembling peptide. The rheological properties of aqueous pharmaceutical compositions containing exemplary self-assembling peptides KLNL12 (SEQ ID NO: 21), KIQI13 (SEQ ID NO: 29), or NLEL12 (SEQ ID NO: 33) were determined by performing frequency sweep tests over the storage modulus.

[0262] Figures 13–15 show the rheological properties of aqueous pharmaceutical compositions containing 1% (w / v) of KLNL12 (SEQ ID NO: 21) (Figure 13), KIQI13 (SEQ ID NO: 29) (Figure 14), or NLEL12 (SEQ ID NO: 33) (Figure 15) at pH 7.5, before and after exposure to buffered Dulbecco's Modified Eagle Medium (DMEM) to simulate body fluids. In each case, the rheological properties (e.g., mechanical strength) of the peptide solution increased after DMEM treatment.

[0263] Figures 16-18 show the rheological properties of aqueous pharmaceutical compositions containing 1% (w / v) KLNL12 (SEQ ID NO: 21) (Figure 16), KIQI13 (SEQ ID NO: 29) (Figure 17), or NLEL12 (SEQ ID NO: 33) (Figure 18) in an isotonic saline (0.9% NaCl) at pH 7.5, before and after exposure to DMEM. The rheological properties (e.g., mechanical strength) of the peptide solutions increased after DMEM treatment.

[0264] The above data demonstrates that aqueous pharmaceutical compositions containing KLNL12 (SEQ ID NO: 21), KIQI13 (SEQ ID NO: 29), or NLEL12 (SEQ ID NO: 33) exhibit increased mechanical strength in the presence of physiological solutions, and are therefore suitable for in vivo therapeutic applications.

[0265] Example 6. A pharmaceutical composition containing an exemplary self-assembling peptide showed reduced tissue damage compared to the pharmaceutical composition containing RADA16. Given that the self-assembling peptides disclosed herein (Table 5; SEQ ID NOs: 21-40) have a non-zero net charge when formulated at a neutral pH (e.g., pH 7.5), it was hypothesized that these compositions would exhibit reduced adverse effects when administered to or in contact with mammalian tissues compared to compositions containing the aforementioned self-assembling peptides (e.g., RADA16) formulated at an acidic pH. Therefore, using an in vivobuta model system, the ability of these compositions to induce submucosal elevation and the degree of tissue damage upon injection into the gastrointestinal submucosa was evaluated.

[0266] In this model, submucosal injection using liquid compositions was performed, and the morphology of the injection site was analyzed. Submucosal injection with aqueous solutions is used during resection of lesions from the gastrointestinal tract to facilitate lesion removal by providing a safety cushion during resection (see, for example, Kim et al. (2013) World J. Gastroenterol. 19(20):3069-76, incorporated herein by reference). Here, aqueous pharmaceutical compositions containing three exemplary self-assembling peptides formulated at pH 7.5: KLNL12 (SEQ ID NO: 21), NLKL12 (SEQ ID NO: 23), or KIQI13 (SEQ ID NO: 29) were compared with aqueous pharmaceutical compositions containing the aforementioned RADA16 self-assembling peptide (formulated at pH 2.5), a 0.4% solution of sodium hyaluronate (MucoUp®, Boston Scientific Japan KK, Tokyo, Japan), and physiological saline, pH 7.5. As shown in Figures 21A and 21B, injection of compositions containing KLNL12 (SEQ ID NO: 21), NLKL12 (SEQ ID NO: 23), or KIQI13 (SEQ ID NO: 29) resulted in submucosal elevation. These compositions showed superior submucosal elevation over time compared to physiological saline and MucoUp®.

[0267] The effects of injection of aqueous pharmaceutical compositions containing RADA16 (SEQ ID NO: 91) at pH 2.5, with and without 0.9% (w / v) NaCl, on the morphology of submucosal tissue were analyzed after tissue dissection at the injection site using an electroknife. As shown in Figures 22A and 22B, injection of compositions containing RADA16 (SEQ ID NO: 91), with and without 0.9% NaCl, resulted in damaged tissue, including mucosal aggregation, which was visualized as a white precipitate near the injection site. In contrast, no tissue damage was observed when physiological saline was injected (Figure 22A).

[0268] The effects of injection of aqueous pharmaceutical compositions containing exemplary self-assembling peptides KLNL12 (SEQ ID NO: 21), NLEL12 (SEQ ID NO: 33), QLEL12 (SEQ ID NO: 35), or LELQ12 (SEQ ID NO: 36), all containing 0.9% (w / v) NaCl at pH 7.5, on the morphology of submucosal tissue were also analyzed after dissection of the injection site using an electroknife. As shown in Figures 23A–23D, mucosal aggregation was not observed at the injection site, and the morphology of the site was easily visible.

[0269] As an additional control, the effects of injection of phosphate-buffered saline (PBS) at pH 7.4 or pH 2.5 on the morphology of submucosal tissue were analyzed after dissection of the injection site using an electroknife. Mucosal aggregation, visualized as a white precipitate near the injection site, was observed only at sites injected with acidic PBS (i.e., pH 2.5) (data not shown), confirming that the changes in mucosal morphology were likely attributable to the pH of the composition.

[0270] Table 8 below summarizes the appearance of the gastric mucosa after injection of each composition used in this example. Table 8. Appearance of the porcine gastric mucosa at the injection site. [Table 8-1] [Table 8-2]

[0271] The data presented in this example demonstrate that injection of an aqueous pharmaceutical composition of the RADA16 formulation (SEQ ID NO: 91) at pH 2.5 into the submucosa of a pig's stomach results in mucosal aggregation after approximately 10–15 minutes. In contrast, injection of aqueous pharmaceutical compositions containing exemplary self-assembling peptides KLNL12 (SEQ ID NO: 21), KIQI12 (SEQ ID NO: 99), NLEL12 (SEQ ID NO: 33), QLEL12 (SEQ ID NO: 35), or LELQ12 (SEQ ID NO: 36), all at pH 7.5, does not appear to alter the submucosal tissue morphology. This result is consistent with the concept that acidic compositions (e.g., those containing RADA16) cause damage to mucosal tissue, while injection of compositions with a neutral pH results in less tissue damage.

[0272] In another embodiment, mucosal aggregation, visualized as a white precipitate near the injection site, was evaluated in vivo in the esophagus of a pig.

[0273] Table 9 below summarizes the appearance of the esophageal mucosa after injection of each composition used in this example.

[0274] Mucosal aggregation was observed at injection sites of KIQI12 (SEQ ID NO: 99), as well as acidic PBS (pH 2) and RADA16 (SEQ ID NO: 91). KIQI12 (SEQ ID NO: 99) is a representative peptide among those positively charged at physiological pH. However, mucosal aggregation was not observed with NLEL12 (SEQ ID NO: 33), QLEL12 (SEQ ID NO: 35), and LELQ12 (SEQ ID NO: 36), which are negatively charged at physiological pH. This result demonstrates that changes in mucosal morphology may be attributable to both the positive or negative charge of the composition at pH and physiological pH in several submucosal tissues, including the esophagus. Table 9. Appearance of the porcine esophageal mucosa at the injection site. [Table 9-1] [Table 9-2]

[0275] Example 7. Pharmaceutical composition with increased mechanical strength In some embodiments, a peptide formulated as an aqueous solution, 700 μl in volume, was placed between rheometer plates (DHR-1, TA Instruments, plates with a 40 mm cone and a 2.0° cone angle measurement shape), and a frequency sweep test was performed at frequencies from 0.1 Hz to 10 Hz and a strain of 0.1%; measurements were performed after a 2-minute relaxation time at 37°C. In some embodiments, the frequency sweep test was performed after exposure to Dulbecco's Modified Eagle Medium (DMEM), where 10 mL of DMEM was added to the chamber surrounding the plate, the sample in the plate was immersed in the DMEM, and the frequency test was performed 20 minutes later as described.

[0276] Figure 24A shows the changes in the rheological properties of an aqueous pharmaceutical composition containing 0.15% (w / v) QLEL12 (SEQ ID NO: 35) with 0.9% NaCl (w / v) at pH 7.5, as reflected by the increase in mechanical strength after exposure to DMEM. At a frequency of 0.1 Hz, the mechanical strength of the composition containing the self-assembling peptide increased 23-fold. At a frequency of 1 Hz, the mechanical strength of the composition containing the self-assembling peptide increased 20-fold. At a frequency of 10 Hz, the mechanical strength of the composition containing the self-assembling peptide increased 9-fold.

[0277] Figure 24B shows the rheological properties of aqueous pharmaceutical compositions containing QLEL12 (SEQ ID NO: 35) with 0.9% NaCl (w / v) at pH 7.5 at various concentrations from 0.1% (w / v) to 0.3% (w / v). A linear relationship was observed between the rheological properties and the concentration of the self-assembling peptide. Increasing the concentration (concertation) of the self-assembling peptide in the composition resulted in increased mechanical strength.

[0278] Example 8: Pharmaceutical composition having submucosal elevation and increased lifting ability In some embodiments, the ability of peptide solutions to rise in the submucosa was evaluated using an exovivoine dog model. For this study, QLEL12 (SEQ ID NO: 35) was selected because it exhibited good rheological properties and did not show any unclear / turbid areas in the submucosa. QLEL12 (SEQ ID NO: 35) solutions at 0.1%, 0.15%, 0.2%, and 0.3% (w / v) concentrations were prepared in 0.9% NaCl at pH 7.5. Physiological saline was used as the reference solution. 0.5 mL of each sample was injected into the submucosa of either the stomach or colon of an exovivoine dog model. The elevation height was measured using a caliper at 0, 10, 20, 30, 40, 50, and 60 minutes after the initial injection. After injecting each sample solution into the submucosa of the stomach and colon using a hypodermic needle, the elevation height was measured using a caliper. The elevation height is the distance between the bottom of the flat surface of the skin after injection into the submucosa and the top of the swollen blister.

[0279] After the solution was injected, its lifting ability to lift the submucosa from the muscular layer was measured. In some embodiments, the muscular layer was the gastrointestinal tract. If the initial lifting height and the overall lifting height over time are sufficient for the submucosal removal procedure, it is considered to have good lifting ability.

[0280] Figure 25A shows the elevation of the submucosal layer in the canine stomach after injection of QLEL12 (SEQ ID NO: 35) containing 0.5 mL of 0.9% NaCl (w / v) at pH 7.5 in various concentrations from 0.1% (w / v) to 0.3% (w / v).

[0281] Figure 25B shows the relative elevation of the canine colon to its initial height after injection of QLEL12 (SEQ ID NO: 35) containing 0.5 mL of 0.9% NaCl (w / v) at pH 7.5 in various concentrations from 0.1% (w / v) to 0.3% (w / v).

[0282] Figure 25C shows the elevation of the submucosa in the canine colon after injection of QLEL12 (SEQ ID NO: 35) containing 0.5 mL of 0.9% NaCl (w / v) at pH 7.5 in various concentrations from 0.1% (w / v) to 0.3% (w / v).

[0283] Figure 25D shows the relative elevation of the canine colon to its initial height after injection of QLEL12 (SEQ ID NO: 35) containing 0.5 mL of 0.9% NaCl (w / v) at pH 7.5 in various concentrations from 0.1% (w / v) to 0.3% (w / v).

[0284] Treatment with compositions containing representative self-assembling peptides resulted in a sustained increase in the height of submucosal elevation in both the colon and stomach for all concentrations tested. Treatment with compositions containing representative self-assembling peptides also resulted in a sustained increase in the relative elevation of submucosal elevation in both the colon and stomach for all concentrations tested. The lifting capacity of compositions containing representative self-assembling peptides also showed good lifting capacity, which helps to facilitate the removal of certain sections of the submucosa.

[0285] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention pertains. Methods and materials are described herein for use in this invention; other suitable methods and materials known in the art may also be used. Materials, methods and examples are illustrative and not intended to limit. All publications, patent applications, patents, sequences, database entries and other references referenced herein are incorporated by reference in their entirety. In case of any conflict, this specification, including definitions, shall prevail.

Claims

1. A composition for forming a hydrogel, comprising a self-assembling peptide consisting of the amino acid sequence shown in SEQ ID NO: 13 or SEQ ID NO:

16.

2. The self-assembling peptide is An N-terminal functional group selected from the group consisting of acetyl, formyl, pyroglutamyl (pGlu), biotin, polyethylene glycol (PEG), urea, alkylamine, carbamate, sulfonamide, dansyl, 2,4-dinitrophenyl, fluorescein, 7-methoxycoumarin acetate, 9-fluorenylmethyloxycarbonyl, palmitic acid, succinyl, chloroacetyl, maleimide, benzyloxycarbonyl, bromoacetyl, nitrilotriacetyl, tert-butoxycarbonyl, 4-hydroxyphenylpropionic acid, allyloxycarbonyl, butyric acid, fatty acids, and trityl, and A C-terminal functional group selected from the group consisting of amines, amides, N-alkylamides, aldehydes, esters, alcohols, para-nitroanilide (pNA), 7-amino-4-methylcoumarin (Amc), hydrazides, hydroxamic acid, chloromethyl ketone, p-nitroaniline, para-nitrophenol, hydroxysuccinimide ester, fluoromethyl ketone, cysteamide, 9-fluorenemethyl (Fm) ester, allyl ester, 2,4-dimethoxybenzyl ester, 2-phenylisopropyl ester, p-nitrobenzyl ester, and 2-chlorotrityl ester. The composition according to claim 1, comprising one or both of the above.

3. The composition according to claim 2, wherein the self-assembling peptide further comprises at least one biologically active peptide motif at the N-terminus, C-terminus, or both of the self-assembling peptide.

4. The composition according to 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. NaCl, KCl, MgCl ２ CaCl ２ NH ４ Cl, Na ２ HPO ４ 7KH ２ PO ４ and CaSO ４ One or more salts selected from the group consisting of; or One or more sugars selected from the group consisting of dextrose, mannitol, glycerin, sucrose, and trehalose. The composition according to claim 2, further comprising an isotonic agent containing the above.

6. The composition according to claim 5, wherein the isotonic agent comprises one or more salts and is present at a concentration of 0.01 M to 0.3 M or 0.15 M.

7. The composition according to claim 5, wherein the isotonic agent contains 1 or more sugars and is present in a concentration of 0.1 to 10% (w / v) or 10% (w / v).

8. The composition according to claim 2, having a pH in the range of 6 to 8.

9. The composition according to claim 8, wherein the net charge of the self-assembling peptide is greater than or equal to +1, or less than or equal to -1.

10. The composition according to claim 2, wherein the concentration of the self-assembling peptide is 0.01% (w / v) to 10% (w / v), 0.1% (w / v) to 5% (w / v), 0.5% (w / v) to 1.5% (w / v), or 1% (w / v).

11. The composition according to claim 2, further comprising isolated cells.

12. The composition according to claim 11, wherein the isolated cells are mammalian cells selected from the group consisting of immune cells, stem cells, chondrocyte progenitor cells, pancreatic progenitor cells, myoblasts, fibroblasts, keratinocytes, nerve cells, glial cells, astrocytic cells, preadipocytes, adipocytes, vascular endothelial cells, endothelial progenitor cells, mesenchymal cells, neural stem cells, smooth muscle progenitor cells, cardiomyocytes, fetal dermal fibroblasts, epidermal keratinocytes, myoblasts, and capillary endothelial cells.

13. The composition according to claim 2, further comprising a bioactive agent.

14. The composition according to claim 13, wherein the bioactive agent is selected from the group consisting of hormones, growth factors, insulin, enzymes, siRNA, shRNA, antisense RNA, antibiotics, antibodies, and anti-inflammatory agents.

15. The composition according to claim 2, which is an aqueous solution.

16. The composition according to claim 2, wherein the hydrogel is formed in vitro or in vivo when the composition is administered.

17. The composition according to claim 16, wherein the hydrogel has a storage modulus of at least 10 Pascals (Pa).

18. A manufactured article comprising the composition described in claim 2, the article being a syringe, vial, automatic infusion device, tube, or catheter.

19. A composition for promoting tissue repair or regeneration in a subject requiring promotion of tissue repair or regeneration, wherein the composition comprises a self-assembling peptide consisting of the amino acid sequence shown in SEQ ID NO: 33 or SEQ ID NO: 36, the composition is administered to the damaged and / or diseased tissue of the subject, thereby promoting tissue repair or regeneration of the tissue, and the composition is characterized by forming a hydrogel upon administration.

20. The composition according to claim 19, wherein the tissue is skin, bone, cartilage, nerve, ligament, tendon, vascular tissue, eye, muscle, or heart tissue.

21. The composition according to claim 19, wherein the subject has a congenital disease or disorder that necessitates tissue repair or regeneration; or the subject suffers from injury that necessitates tissue repair or regeneration, the injury being the result of surgery, trauma, stroke, tumor, or disease or disorder.

22. A composition for promoting wound healing in a subject requiring the promotion of wound healing, wherein the composition comprises a self-assembling peptide consisting of an amino acid sequence shown in SEQ ID NO: 33 or SEQ ID NO: 36, wherein the composition is administered to a wound of the subject and thereby promotes wound healing and / or antibacterial activity, wherein the wound includes abrasions, burns, cracks, crushes, cuts, ulcers, lacerations, incisions or scratches, and the composition forms a hydrogel upon administration.

23. A composition for reducing bleeding at a site within a target, wherein the composition comprises a self-assembling peptide consisting of an amino acid sequence shown in SEQ ID NO: 33 or SEQ ID NO: 36, and the composition is administered to the site, thereby creating a physical barrier by forming a hydrogel, and thereby reducing bleeding at the site within the target.

24. A composition for use in a method for excising a lesion from a target site of the gastrointestinal tract, wherein the composition comprises a self-assembling peptide consisting of the amino acid sequence shown in SEQ ID NO: 33 or SEQ ID NO: 36, and the method is A step of administering a sufficient amount of the composition to lift the lesion into the submucosa beneath the lesion, wherein the composition forms a hydrogel upon administration; The steps of removing the lesion from the aforementioned part of the gastrointestinal tract of the subject and A composition containing the following:

25. The composition according to claim 24, wherein the lesion includes a polyp, ulcer, or tumor.

26. The composition according to claim 24, wherein the lesion is located in a region of the gastrointestinal tract selected from the group consisting of the mouth, throat, esophagus, stomach, small intestine, large intestine, colon, and rectum.

27. A method for culturing cells, comprising the step of bringing the cells into contact with the composition described in claim 2.

28. A composition for use in a method for treating target alveolar cysts, wherein the composition comprises a self-assembling peptide consisting of the amino acid sequence shown in SEQ ID NO: 33 or SEQ ID NO: 36, and the method is The steps include introducing the delivery device into the target region of the alveolar cyst of the target; The steps include positioning the end of the delivery device in the target region where treatment of the alveolar cysts is desired; The steps include: administering an effective amount and effective concentration of the composition to the target region through the delivery device to treat the alveolar cysts by forming a hydrogel barrier under physiological conditions in the target region; The steps of removing the delivery device from the target region; A step of disintegrating the alveolar cysts after administering the composition. A composition containing the following:

29. A composition for use in a method for filling a target interosseous space, wherein the composition comprises a self-assembling peptide consisting of the amino acid sequence shown in SEQ ID NO: 33 or SEQ ID NO: 36, and the method is Steps include: introducing the delivery device into the target bone; The steps include positioning the end of the delivery device proximal to the interstitial space of the bone where bone growth promotion is desired; The steps include: administering the composition through the delivery device in a concentration sufficient to form a hydrogel scaffold under physiological conditions; The step of removing the delivery device A composition containing the following:

30. A composition for treating a target dry eye, wherein the composition comprises a self-assembling peptide consisting of an amino acid sequence shown in SEQ ID NO: 33 or SEQ ID NO: 36, and the composition is characterized by being administered to the target eye, wherein the composition forms a hydrogel upon administration.

31. A self-assembling peptide consisting of the amino acid sequence shown in SEQ ID NO: 33 or SEQ ID NO: 36.

Images