Vascular embolization system

Abstract

To provide a vascular embolization system.SOLUTION: A system and method for blocking biological vessel is provided. The above system and method may include a step of introducing an amphipathic peptide into the vessel. The peptide may comprise at least 13 amino acids, which may alternate between hydrophobic and hydrophilic amino acids. The peptide can naturally form a β-sheet in aqueous solution in the presence of cation. In one embodiment, the method comprises a step of introducing a catheter into a biological vessel and a step of placing the distal end of the catheter in a target region of a biological vessel where at least partial occlusion is desired.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] (Sequence Listing) This application contains a Sequence Listing that has been submitted electronically in ASCII format, which is incorporated herein by reference in its entirety. The ASCII copy was created on November 14, 2013, is named T2071-7000WO_SL.txt, and is 23,527 bytes in size. [Background technology]

[0002] FIELD OF THE DISCLOSURE The present disclosure relates to macroscopic membranes that can be used in medical, research, and industrial applications. More specifically, the present disclosure relates to membranes, hydrogels, compositions, and solutions that can be used in vascular embolization systems and embolization procedures. Vascular embolization systems can provide an approach to at least partially block biological pathways or channels, including vessels, veins, portal veins, arteries, and tubes that can transport blood and other fluids (e.g., lymphatic fluid). Summary of the Invention [Means for solving the problem]

[0003] (Summary) A method is provided for blocking a biological vessel in a subject, the method comprising introducing a catheter into the biological vessel and positioning a distal end of the catheter at a target area of ​​the biological vessel where at least partial occlusion is desired. The method further comprises administering through the catheter a solution comprising an amphipathic peptide comprising at least 12 amino acids of alternating hydrophobic and hydrophilic amino acids in an amount and at a concentration effective to form a hydrogel under physiological conditions to enable at least partial blocking of the biological vessel. The method further comprises removing the catheter from the biological vessel with the at least partial occlusion in place.

[0004] A kit is provided for blocking a biological vessel in a subject, the kit comprising a solution comprising an amphipathic peptide comprising at least 12 amino acids of alternating hydrophobic and hydrophilic amino acids in an amount and at a concentration effective to form a hydrogel under physiological conditions to enable at least partial blocking of the biological vessel, the kit further comprising instructions for administering the solution to the biological vessel in the subject.

[0005] A method is provided for promoting the blocking of a biological vessel in a subject, the method comprising providing a solution comprising an amphipathic peptide comprising at least 12 amino acids of alternating hydrophobic and hydrophilic amino acids in an amount and at a concentration effective to form a hydrogel under physiological conditions to enable at least partial blocking of the biological vessel, the method further comprising providing instructions for administering the solution to a target area of ​​the biological vessel by introducing the solution into a catheter disposed in the biological vessel. In an embodiment of the present invention, for example, the following items are provided: (Item 1) 1. A method for blocking a biological vessel in a subject, the method comprising: introducing the catheter into the biological vessel; positioning a distal end of the catheter at a target area of ​​the biological vessel where at least partial occlusion is desired; administering through the catheter a solution comprising an amphipathic peptide comprising at least 12 amino acids of alternating hydrophobic and hydrophilic amino acids in an amount and at a concentration effective to form a hydrogel under physiological conditions to permit at least partial obstruction of the biological vessel; removing the catheter from the biological vessel with the at least partial occlusion in place; The method includes: (Item 2) 2. The method of claim 1, wherein the peptide solution comprises an imaging agent. (Item 3) 3. The method of claim 2, further comprising visualizing a local area comprising at least a portion of the biological vessel. (Item 4) The step of visualizing the area including at least a portion of the biological vessel includes the steps of: identifying the target region of the biological vessel; introducing said catheter; placing the distal end of the catheter at the target area; administering said solution; removing the catheter; and visualizing the biological vessel after removing the catheter. 4. The method according to claim 3, further comprising visualizing the area during at least one of the steps of: (Item 5) 5. The method according to claim 4, wherein the step of visualizing the local area comprises imaging using X-ray photography. (Item 6) 4. The method of claim 3, wherein visualizing the local area provides for selective administration of the solution to the biological vessel. (Item 7) 4. The method of claim 3, further comprising visualizing the area for a period of about two weeks after the administration. (Item 8) 2. The method of claim 1, wherein at least one of the effective amount and the effective concentration is based in part on a diameter of the target region of the biological vessel. (Item 9) 2. The method of claim 1, wherein at least one of the effective amount and the effective concentration is based in part on a flow rate of blood in the biological vessel. (Item 10) 2. The method of claim 1, wherein at least one of the effective amount and the effective concentration is based in part on providing nanofibers of the hydrogel having an average pore size smaller than an average diameter of red blood cells of the subject. (Item 11) 2. The method of claim 1, wherein the concentration effective to enable at least partial blocking of the biological vessel comprises a concentration in the range of about 0.1 weight / volume (w / v)% to about 3 w / v% peptide. (Item 12) 2. The method of claim 1, wherein the amount effective to allow at least partial blocking of the biological vessel comprises a volume ranging from about 0.1 mL to about 5 mL. (Item 13) 2. The method of claim 1, further comprising monitoring the area around the at least partial blockage to determine effectiveness of the at least partial blockage. (Item 14) 2. The method of claim 1, wherein the blockage formed is used in the treatment of a lesion, malformation, or congenital disease in a biological vessel. (Item 15) 15. The method of claim 14, wherein the blockage formed is used in the treatment of one of a patent ductus arteriosus (PDA) and a major aortopulmonary collateral artery (MAPCA). (Item 16) 15. The method of claim 14, wherein the blockage formed is used in the treatment of a disorder, malformation, or congenital disease selected from the group consisting of recurrent hemoptysis, arteriovenous malformation, cerebral aneurysm, gastrointestinal bleeding, epistaxis, postpartum bleeding, intraoperative bleeding, and uterine fibroids. (Item 17) 2. The method of claim 1, wherein the blockage formed is used in reducing cancer cells. (Item 18) 2. The method of claim 1, wherein the peptide solution is substantially free of cells. (Item 19) 2. The method of claim 1, wherein the peptide solution is substantially free of drugs. (Item 20) 2. The method of claim 1, wherein the subject is a mammal. (Item 21) 21. The method of claim 20, wherein the subject is a human. (Item 22) 2. The method of claim 1, wherein administering the solution comprises administering the solution in a single dose. (Item 23) 2. The method of claim 1, wherein administering the solution comprises administering the solution in at least two doses. (Item 24) 2. The method of claim 1, wherein the peptide has one of the amino acid sequences RADARADARADARADA (SEQ ID NO: 7), IEIKIEIKIEIKI (SEQ ID NO: 8), and IEIKIEIKIEIKIEIKI (SEQ ID NO: 9). (Item 25) 2. The method of claim 1, further comprising the step of evaluating the subject to determine the need for blocking a biological vessel and preparing the peptide solution. (Item 26) 26. The method according to claim 25, wherein the step of preparing the peptide solution comprises adding an imaging agent to a pre-solution containing the peptide. (Item 27) 2. The method of claim 1, wherein the solution is administered to allow complete blocking of the biological vessel. (Item 28) 2. The method of claim 1, further comprising the step of introducing a guidewire into the biological vessel prior to introducing the catheter. (Item 29) 1. A kit for blocking a biological vessel in a subject, the kit comprising: a solution comprising an amphipathic peptide comprising at least 12 amino acids of alternating hydrophobic and hydrophilic amino acids in an amount and at a concentration effective to form a hydrogel under physiological conditions to permit at least partial obstruction of said biological vessel; and instructions for administration of the solution to the biological vessel in the subject; Including the kit. (Item 30) 30. The kit of claim 29, further comprising a catheter for introducing the solution into the biological vessel of the subject. (Item 31) 30. The kit of item 29, further comprising instructions for adding a contrast agent to the solution in an amount suitable for visualizing the step of administering the solution. (Item 32) 32. The kit of item 31, further comprising at least one of a contrast agent and a sucrose solution. (Item 33) 30. The kit of claim 29, further comprising instructions for diluting the solution to administer an effective concentration of the solution to the biological vessel in the subject. (Item 34) 34. The kit of claim 33, further comprising instructions for determining the effective concentration of the solution to the biological vessel in the subject based on a diameter of the biological vessel in a target area. (Item 35) 1. A method for promoting blocking of a biological vessel in a subject, the method comprising: providing a solution comprising an amphipathic peptide comprising at least 12 amino acids of alternating hydrophobic and hydrophilic amino acids in an amount and at a concentration effective to form a hydrogel under physiological conditions to enable at least partial obstruction of said biological vessel; and providing instructions for administering the solution to a target region of the biological vessel via introduction of the solution into a catheter positioned in the biological vessel; The method includes: (Item 36) 36. The method of claim 35, further comprising providing instructions for adding a contrast agent to the solution. (Item 37) 37. The method of claim 36, further comprising providing instructions for visualizing an area comprising at least a portion of the biological vessel. (Item 38) The step of providing instructions for visualizing a local area including at least a portion of the biological vessel includes the steps of: identifying a target region of said biological vessel; introducing a catheter; positioning a distal end of the catheter at the target area; administering said solution; removing the catheter from the biological vessel with the at least partial blockage in place; and visualizing the biological vessel after removing the catheter; 38. The method of claim 37, further comprising providing instructions for visualizing the area during at least one of the steps. (Item 39) 40. The method of claim 38, wherein the step of providing instructions for visualizing the area comprises imaging using X-ray photography. (Item 40) 38. The method of claim 37, further comprising providing instructions for visualizing the area for a period of about two weeks after said administration. (Item 41) 36. The method of claim 35, comprising providing instructions for adjusting at least one of the effective amount and the effective concentration based in part on a diameter of the target region of the biological vessel. (Item 42) 36. The method of claim 35, wherein at least one of the effective amount and the effective concentration is based in part on a flow rate of blood in the biological vessel. (Item 43) 36. The method of claim 35, wherein at least one of the effective amount and the effective concentration is based in part on providing a hydrogel having an average pore size smaller than an average diameter of red blood cells of the subject. (Item 44) 36. The method of claim 35, wherein the concentration effective to enable at least partial blocking of the biological vessel comprises a concentration ranging from about 0.1% to about 3% by weight of peptide. (Item 45) Item 36. The method of item 35, wherein the amount effective to allow at least partial blocking of the biological vessel comprises a volume ranging from about 0.1 mL to about 5 mL. (Item 46) 36. The method of claim 35, further comprising monitoring the area around the at least partial blockage to provide indications for determining cell necrosis. (Item 47) 36. The method of claim 35, wherein the blockage formed is used in the treatment of a lesion, malformation, or congenital disease in a biological vessel. (Item 48) 36. The method of claim 35, wherein the blockage formed is used in reducing cancer cells. (Item 49) 36. The method of claim 35, wherein the peptide solution is substantially free of cells. (Item 50) 36. The method of claim 35, wherein the peptide solution is substantially free of drugs. (Item 51) 36. The method of claim 35, wherein the subject is a mammal. (Item 52) 52. The method of claim 51, wherein the subject is a human. (Item 53) 36. The method of claim 35, wherein the peptide has one of the amino acid sequences RADARADARADARADA (SEQ ID NO: 7), IEIKIEIKIEIKI (SEQ ID NO: 8), and IEIKIEIKIEIKIEIKI (SEQ ID NO: 9). [Brief description of the drawings]

[0006] The accompanying drawings are not intended to be drawn to scale and, for purposes of clarity, not all components may be shown. [Figure 1] FIG. 1 is a cross-sectional image of a portal vein embolization using a peptide solution of the present disclosure. [Diagram 2] Figure 2 is an angiogram of a normal hepatic artery. [Diagram 3] FIG. 3 is an image of an injection of a substance of the present disclosure. [Figure 4] FIG. 4 is an angiogram of the hepatic artery after injection of a substance of the present disclosure. [Diagram 5]FIG. 5 is an angiogram of the hepatic artery two weeks after injection of the material of the present disclosure. [Figure 6A] FIG. 6A is a histopathological image of a peptide hydrogel located in the hepatic artery. [Figure 6B] FIG. 6B is a histopathological image of the peptide hydrogel located in the hepatic artery. [Figure 6C] FIG. 6C is a histopathological image of the peptide hydrogel located in the hepatic artery. [Figure 7] FIG. 7 shows images of hepatocyte necrosis at the site of peptide hydrogel embolization. [Figure 8] FIG. 8 is an image of the artery before embolization. [Figure 9] FIG. 9 is an image of the artery after embolization. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0007] (Detailed Description) Embolization is a procedure that creates a blockage, lodging, obstruction, or embolization in one or more biological pathways or channels. The biological pathways or channels may include vessels, veins, portal veins, arteries, and tubes that may transport blood and other fluids (e.g., lymphatic fluid). Embolization is used to treat a wide variety of conditions that affect various organs of a subject's body, including the human body. One or more of the vessels may be targeted to intentionally prevent or reduce blood circulation to a desired target. The embolization procedure may be used to treat tumors or From other pathological processes Those blood donors Salary remove (perfusion) Embolization may be used to intentionally create such a blockage, retention, or occlusion in order to prevent or reduce the risk of infection. Embolization may be used to treat disorders, malformations, or congenital diseases in biological vasculature. For example, embolization may be used to treat patent ductus arteriosus (PDA). The embolization procedure may be used to treat major aortopulmonary collateral arteries (MAPCAs), recurrent hemotysis, arteriovenous malformations, cerebral aneurysms, gastrointestinal bleeding, epistaxis, postpartum bleeding, intraoperative bleeding, and uterine fibroids.

[0008] Embolization can be accomplished by a number of different techniques. Embolization can be accomplished by administering a substance, such as a liquid, to a desired or predetermined location (e.g., a target area). Administration can include applying or injecting a substance (e.g., a liquid) to the desired or predetermined location (e.g., a target area).

[0009] Embolization can be used to close or block all or part of the vessels that form the aneurysm to prevent the aneurysm from rupturing. Embolization can also be used to close or block all or part of certain blood vessels around the site of a subject undergoing surgery, for example, during surgery for a cerebral arteriovenous malformation (AVM).

[0010] Obstructing materials can be used to create a blockage or occlusion in a biological vessel to achieve embolization. Embolic materials can include metal coils, collagen, cyanoacrylate, and other materials. The materials can be inserted or placed using a catheter at the desired surgical site. Balloons can also be implanted in the target vessel and filled with saline.

[0011] Metal coils may remain permanently in vivo, but the safety of applying these coils long term is unknown. The metal coils may also cause incompatibility with magnetic devices. Collagen may have biological incompatibility, and cyanoacrylate may be toxic in vivo.

[0012] Specific liquid embolization agents may include onyx, n-butyl-2-cyanoacrylate (nbca), and ethiodol (iodine, made from poppy seed oil). Schlerosing agents, which harden the endothelial layer of blood vessels, may also be used. Examples of such agents include ethanol, ethanolamine oleate, and sotradecol. Particulate embolization agents include polyvinyl alcohol and acrylic gelatin microspheres.

[0013] The present disclosure provides methods for embolizing or blocking biological vessels, methods for facilitating blocking biological vessels, and kits for use in blocking biological vessels. Biological vessels may include blood vessels and lymphatic vessels. Blood vessels may include arteries, veins, portal veins, and capillaries. The term vascular may refer to biological vessels, including arteries, veins, portal veins, capillaries, and tubes.

[0014] The method may include a step of blocking or occluding a biological vessel in a subject, or a method for promoting blocking or occluding a biological vessel in a subject. As used herein, the term "subject" is intended to include humans and non-human animals (e.g., vertebrates, large animals, and primates). In certain embodiments, the subject is a mammalian subject, and in certain embodiments, the subject is a human subject. Although human applications are clearly envisioned, vertebrate applications (e.g., non-human animal applications) are also envisioned herein. The term "non-human animal" of the present invention includes all vertebrates, such as non-mammals (e.g., birds, e.g., chickens; amphibians; reptiles) and mammals (e.g., non-human primates, domesticated animals, and agriculturally useful animals, such as sheep, dogs, cats, cows, pigs, rats, among others).

[0015] The embolization, blockage, or occlusion may be partial or complete. Complete means that the embolization, blockage, or occlusion prevents substantially all blood flow after the embolization, blockage, or occlusion. The systems and methods may include administration, application, or injection of a self-assembling peptide or a solution containing a self-assembling peptide to a predetermined or desired target area. The self-assembling peptide may be applied or introduced into a biological vessel in the form of a self-assembling peptide solution, hydrogel, membrane, or other form.

[0016] The self-assembling peptide solution can be an aqueous self-assembling peptide solution. The self-assembling peptide (also referred to herein as a "peptide" or an "amphiphilic peptide") can be administered, applied, or injected in a solution that is substantially free of cells. In certain embodiments, the self-assembling peptide can be administered, applied, or injected in a solution that is free of cells.

[0017] The self-assembling peptides may also be administered, applied, or injected in a solution that is substantially free of drugs. In certain embodiments, the self-assembling peptides may be administered, applied, or injected in a solution that is substantially free of drugs. In certain other embodiments, the self-assembling peptides may be administered, applied, or injected in a solution that is substantially free of cells and substantially free of drugs. In yet still other embodiments, the self-assembling peptides may be administered, applied, or injected in a solution that is substantially free of cells and substantially free of drugs.

[0018] The administration of the solution may include, consist of, or consist essentially of a solution comprising, consisting of, or consisting essentially of an amphipathic peptide comprising, consisting of, or consisting essentially of at least 12 amino acids alternating hydrophobic and hydrophilic amino acids.

[0019] The systems and methods may include administering the self-assembling peptide to a given or desired target as a hydrogel, a term that may refer to a colloidal gel dispersed in water. The systems and methods may also include applying the self-assembling peptide to a given or desired target as a solution (e.g., an aqueous peptide solution).

[0020] When the term "administering" is used, it is intended to include, but is not limited to, applying, introducing or injecting the self-assembling peptide in one or more of a variety of forms, including, but not limited to, alone, via a solution (e.g., an aqueous solution), or via a hydrogel, with or without additional components.

[0021] A method for blocking a biological vessel in a subject may include introducing a syringe, pipette, catheter, or other needle-based device into the biological vessel. The self-assembling peptide may be administered into the biological vessel by a syringe, pipette, catheter, or other needle-based device. The gauge of the syringe needle may be selected to provide an appropriate flow of liquid from the syringe to the target area. This may be based in some embodiments on at least one of the amount of self-assembling peptide or peptide solution administered, the concentration of the peptide in solution, and the viscosity of the peptide solution.

[0022] The method for blocking the biological vessel in the subject may include introducing a catheter into the biological vessel and positioning an end of the catheter in a target area of ​​the biological vessel where at least partial occlusion is desired. The self-assembling peptide may be administered by catheter to the target area of ​​the biological vessel where at least partial occlusion is desired. The use of a catheter may provide more selective administration of the peptide to provide more precise delivery to the target area. Selective administration of the peptide may allow for enhanced and more targeted delivery of the peptide solution so that the blockage of the biological vessel is successful and located in a precise manner at the desired location. The selective administration may provide enhanced and targeted delivery that significantly improves the placement and effectiveness of the blockage in the biological vessel through the use of a syringe or other means.

[0023] The use of a catheter may include the use of ancillary devices, such as a guidewire used to guide the catheter to its location. A guidewire may be introduced into the biological vessel prior to introducing the catheter. Once administration of the peptide solution is completed, or once at least a partial occlusion or blockage is in place, the catheter may be removed from the biological vessel.

[0024] Use of a syringe, needle, pipette, other needle-based device, or catheter may require determining the diameter of the biological vessel to be targeted so that at least a portion of the syringe, needle, pipette, other needle-type device, or catheter can enter the biological vessel to administer the peptide, peptide solution, or hydrogel to the target area.

[0025] In certain embodiments, the hydrogel may be formed in vitro and administered to a desired location in vivo. In certain instances, the location may be the area where it is desired to create an embolism. In other instances, the location may be upstream or downstream of the area where it is desired to form an embolism. In this case, it may be desirable to allow the hydrogel to move or migrate on its own to the area where it is desired to form an embolism. Alternatively, a separate procedure may place the hydrogel in the area where it is desired to form an embolism. The desired location or target area may be part of a biological vessel. The desired location or target area may be part within a biological vessel.

[0026] In certain aspects of the present disclosure, the hydrogel can be formed in vivo. A solution (e.g., an aqueous solution) containing the self-assembling peptide can be inserted into a location or area in a subject in vivo to allow an embolism to be created at that location. In certain instances, the hydrogel can be formed at one location in vivo and allowed to move on its own to the area where it is desired to form an embolism. Alternatively, another procedure can place the hydrogel in the area where it is desired to form an embolism. The peptide of the present disclosure can be in the form of a powder, solution, gel, etc. Since the self-assembling peptide gels in response to changes in solution pH and salt concentration, it can be distributed as a liquid that gels when it comes into contact with a subject during application or administration.

[0027] Certain self-assembling peptides of the present disclosure may provide improved adhesion to tissues than other agents that may be used to embolize biological vessels. Improved adhesion may be due to the composition of the peptide (e.g., the specific amino acids of the peptide), the structure of the peptide once self-assembled, or due to the self-assembly process itself. In certain embodiments, it may be useful in procedures to remove excess bodily fluids (e.g., blood or bile) from a target site or area where it is desired to provide a hydrogel for occlusion.

[0028] In some embodiments, the peptide or hydrogel may be adherent to the tissue or biological vessel. When the peptide or peptide solution is administered, it comes into contact with blood or other fluids in the biological vessel, thereby causing gelation inside the vessel. The peptide solution may move within the vessel, but once it starts to gel, it loses its fluidity and remains in place at a location or target area in the biological vessel. The peptide in the form of a hydrogel may remain in place without adhering to the tissue or biological vessel.

[0029] The present disclosure relates to aqueous solutions, hydrogels, and membranes comprising self-assembling peptides (sometimes referred to as self-assembling oligopeptides, or amphipathic peptides). The peptides may be comprised of amphipathic peptides having from about 6 to about 200 amino acid residues of alternating hydrophilic and hydrophobic amino acids. The self-assembling peptides may exhibit a β structure in aqueous solution at physiological pH and / or in the presence of cations (e.g., monovalent cations).

[0030] The order of availability of the above monovalent cations is Li + >Na + >K + >Cs + It seems like. Cs + The availability of the monovalent cations can be inversely related to the crystal radius of the ion: Li + (0.6 Å), Na + (0.95Å), K + (1.33 Å), and Cs + (1.69 Å) (Pauling, 1960). The correlation also gives the hydration radius of the above ion: Li + (3.4Å), Na + (2.76 Å), K + (2.32 Å), and Cs +(2.28 Å) and the order of enthalpy of the monovalent cations (Pauling, 1960). The presence of the monovalent metal cations may act as catalysts or may be incorporated into the membrane. The size of the filaments (10-20 nm) and the distance between the filaments (50-80 nm) in some of the membranes formed may suggest that hydrated ions may stabilize intermolecular interactions. Some anions (divalent anions, acetate, Cl) - , S.O. 4 -2 , and P.O. 4 -2 ) and organic ions, NH 4 + and Tris-Cl may not induce membrane formation.

[0031] Concentrations of monovalent metal cations (NaCl) as low as 5 mM and as high as 5 M have been found to induce membrane formation within minutes in certain embodiments. Thus, membrane formation may be independent of salt concentration over this wide range. Salt concentrations below 5 mM may also induce membrane formation, but at a slower rate.

[0032] The peptides can generally be stable in aqueous solutions and can self-assemble into large, extremely stable macroscopic structures or matrices when exposed to physiological conditions or salt levels. The presence of monovalent alkali metal ions (e.g., sodium and potassium ions present at physiological levels) promotes the formation of hydrogels from the peptide solutions. Once the hydrogel is formed, it may not degrade under common protein denaturing conditions (e.g., high temperature) or denaturants (e.g., acid, alkali, proteases, urea, guanidine hydrochloride, etc.). Self-assembled peptides can form sheet-like or fibril structures that are visible to the naked eye and have high tensile strength when stained with the dye Congo Red. These materials are substantially resistant to changes in pH, heat, and enzymatic proteolysis. The self-assembled peptides have a fibrous microstructure with small pores as revealed by electron microscopy.

[0033] Physiological conditions may be essentially those that exist for a particular organ or cell system, which may be contrasted with artificial laboratory conditions. The conditions may include one or more characteristics (e.g., one or more particular characteristics or one or more ranges of characteristics). For example, the physiological conditions may include a temperature or temperature range, a pH or pH range, a pressure or pressure range, and one or more concentrations of particular compounds, salts, and other components. For example, in some examples, the physiological conditions may include a temperature in the range of about 20° C. to about 40° C. In some examples, the atmospheric pressure may be about 1 atm. The pH may be in the range of about 6 to about 8. The physiological conditions may include cations, such as monovalent metal cations, that may induce membrane formation. These may include sodium chloride (NaCl). The physiological conditions may also include a glucose, sucrose, or other sugar concentration between about 1 mM and about 20 mM.

[0034] The self-assembling peptides of the present disclosure may have at least 8 amino acids, at least 12 amino acids, or at least 16 amino acids. The peptides may also be complementary and structurally compatible. Complementary refers to the ability of the peptides to interact with each other through ionized pairs and / or hydrogen bonds formed between their hydrophilic side chains, and structurally compatible refers to the ability of complementary peptides to maintain a certain distance between their peptide backbones. Peptides with these properties participate in the intermolecular interactions that result in the formation and stabilization of beta sheets at the secondary structure level and entangled filaments at the tertiary structure level.

[0035] Both homogeneous and heterogeneous mixtures of peptides characterized by the above-mentioned properties can form stable macroscopic films, filaments, and hydrogels. Peptides that are self-complementary and self-compatible can form films in homogeneous mixtures. Heterogeneous peptides (including those that cannot form films in homogeneous solutions but are complementary and / or structurally compatible with each other) can also self-assemble into macroscopic films, filaments, and hydrogels.

[0036] Macroscopic films, filaments, and hydrogels formed from the self-assembling peptides can be stable in aqueous solutions, in serum, and in ethanol, and can be highly resistant to degradation by heat, alkaline and acidic pH (stable at pH 1.5-11), chemical denaturants (e.g., guanidine-HCl, urea, and sodium dodecyl sulfate), and proteases in vitro (e.g., trypsin, α-chymotrypsin, papain, proteinase K, and pronase). They can be non-cytotoxic.

[0037] The above methods and methods for promoting the present disclosure may include administering via a catheter, and providing instructions therefor, a solution comprising an amphipathic peptide comprising at least 12 amino acids of alternating hydrophobic and hydrophilic amino acids in an amount and at a concentration effective to form a hydrogel under physiological conditions to enable at least partial blockage of a biological vessel.

[0038] The method for promoting the above may include providing a solution comprising an amphipathic peptide comprising at least 12 amino acids of alternating hydrophobic and hydrophilic amino acids in an amount and at a concentration effective to form a hydrogel under physiological conditions to enable at least partial blockage of the biological vessel.

[0039] The method and the method for facilitating may include adding contrast agent to the peptide solution or providing instructions for adding contrast agent to the solution. Alternatively, the peptide solution may be manufactured with contrast agent. The contrast agent may provide visual image during the use of X-ray techniques, such as fluoroscopy or angiography. Non-ionic radiopaque contrast medium may include, for example, water-soluble iodine-based solution. The water-soluble iodine-based solution may be iopamidol.

[0040] The method and method for promoting the present disclosure may include visualizing an area including at least a portion of the biological vessel or providing instructions for visualizing an area including at least a portion of the biological vessel. The visualization may occur during at least one of the steps of identifying the target area, introducing the catheter, positioning the distal end of the catheter in the target area, administering the solution, and observing the biological vessel after removing the catheter.

[0041] The visualization step can be achieved by imaging using X-ray photography. The method and the method for promoting can include using X-ray photography to visualize the area or provide instructions to visualize it. The visualization step can occur for a period of time after administering the peptide or removing the catheter. For example, it can occur for up to 5 minutes or 1 hour after administering the peptide or removing the catheter. Visualization can also occur after one or more predetermined intervals. For example, visualization can occur about 24 hours, about 1 week, about 2 weeks, or about 4 weeks after administering the peptide or removing the catheter. Visualization can occur about 3 months later. Visualization can also occur about 6 months later. Instructions can be provided to visualize the area at any one or more of the time periods disclosed herein and for any period of time. For example, at 1 week, the visualization can occur for 1 minute or 5 minutes. At 4 weeks, the visualization can occur for 10 minutes or 3 minutes.

[0042] Visualizing or monitoring the area around the formed blockage may also occur over a period of time or at one or more predetermined intervals after administering the peptide or removing the catheter to determine any one or more of the effectiveness of the blockage, any degradation of the blockage, and any cell or tissue necrosis.

[0043] The disclosed method may further include evaluating the subject to determine the need for biological vascular blockage and preparing the peptide solution. The step of preparing the peptide solution may include adding a contrast agent to a pre-solution containing the peptide.

[0044] The method for facilitating may further include providing instructions for adding a contrast agent to the solution. The method for facilitating may include providing instructions for combining a sufficient amount or volume of the contrast agent to properly at least one of: identify the target area, introduce a catheter or other administration device, place the end of the catheter in the target area, administer the peptide solution, remove the catheter or other administration device, and observe the biological vessel after removing the catheter. The use of the contrast agent may allow visualization of the area where the peptide, peptide solution, or hydrogel is administered.

[0045] The amino acids of the self-assembling or amphiphilic peptides can be selected from d-amino acids, l-amino acids, or combinations thereof. The hydrophobic amino acids include Ala, Val, Ile, Met, Phe, Tyr, Trp, Ser, Thr, and Gly. The hydrophilic amino acids can be basic amino acids (e.g., Lys, Arg, His, Orn); acidic amino acids (e.g., Glu, Asp); or hydrogen bond forming amino acids (e.g., Asn, Gln). Acidic and basic amino acids can be clustered on the peptide. The carboxyl and amino groups of the terminal residues can be protected or unprotected. Membranes can be formed in a homogeneous mixture of self-complementary and self-compatible peptides, or in a heterogeneous mixture of peptides that are complementary and structurally compatible with each other. Peptides that meet the above criteria can self-assemble into macroscopic membranes under appropriate conditions as described herein.

[0046] Peptides of the present disclosure may include peptides having repeating sequences of arginine, alanine, aspartic acid, and alanine (Arg-Ala-Asp-Ala (RADA) (SEQ ID NO: 1)), such peptide sequences are represented by the formula: (RADA) P(where p=2-50) (SEQ ID NO:2). Other peptide sequences can be represented by self-assembling peptides having repeating sequences of isoleucine, glutamic acid, isoleucine and lysine (Ile-Glu-Ile-Lys (IEIK) (SEQ ID NO:3)), such peptide sequences being represented by the formula: (IEIK) P (where p=2-50) (SEQ ID NO: 4). Other peptide sequences can be represented by self-assembling peptides having repeating sequences of lysine, leucine, aspartic acid, and leucine (Lys-Leu-Asp-Leu (KLDL) (SEQ ID NO: 5)), such peptide sequences being represented by the general formula (I): P (where p=2-50) (SEQ ID NO: 6). The self-assembling peptide can be comprised of about 8 to about 200 amino acid residues. In certain embodiments, about 8 to about 32 residues can be used in the self-assembling peptide, while in other embodiments, the self-assembling peptide can have about 12 to about 17 residues. The peptide can have a length of about 5 nm.

[0047] An exemplary self-assembling peptide according to the invention has the sequence Arg-Ala-Asp-Ala-Arg-Ala-Asp-Ala-Arg-Ala-Asp-Ala-Arg-Ala-Asp-Ala (RADA). 4 (SEQ ID NO: 7), the self-assembling peptide RADA16 having the sequence Ile-Glu-Ile-Lys-Ile-Glu-Ile-Lys-Ile-Glu-Ile-Lys-Ile (IEIK) 3 IEIK13, a self-assembling peptide having the sequence Ile-Glu-Ile-Lys-Ile-Glu-Ile-Lys-Ile-Glu-Ile-Lys-Ile-Glu-Ile-Lys-Ile-Glu-Ile-Lys-Ile (IEIK) 4 IEIK17, a self-assembling peptide having the sequence Lys-Leu-Asp-Leu-Lys-Leu-Asp-Leu-Lys-Leu-Asp-Leu (KLDL) 3 The self-assembling peptide KLDL12 may have the following sequence: (SEQ ID NO: 10). (RADA) 4A 1% aqueous solution of (SEQ ID NO: 7) was purchased from 3D-Matrix Co., Ltd. TM Available as PuraMatrix TM is an underwater array (RADA) 4 (SEQ ID NO: 7).

[0048] Certain peptides may contain sequences similar to the cell adhesion ligand RGD (arginine-glycine-aspartic acid). The suitability of these peptides to support cell growth in vitro was tested by introducing into various cultured primary and transformed cells the homopolymeric sheets Ala-Glu-Ala-Glu-Ala-Lys-Ala-Lys-Ala-Glu-Ala-Glu-Ala-Lys-Ala-Lys (AEAEAKAKAEAEAKAK (EAK16) (SEQ ID NO: 11), RAD16 (SEQ ID NO: 22), RADA16 (SEQ ID NO: 7), and a heteropolymer of RAD16 (SEQ ID NO: 22) and EAK16 (SEQ ID NO: 11). The RAD-based peptides may be of particular interest due to the similarity of this sequence to RGD. The RAD sequence is a high affinity ligand present in the extracellular matrix protein tenascin, which is recognized by integrin receptors. The EAK16 peptide (SEQ ID NO: 11) and other peptides disclosed herein were derived from portions of the yeast protein zuotin.

[0049] The self-assembly of the peptides can be attributed to hydrogen bonds and hydrophobic bonds between the peptide molecules of the amino acids that make up the peptide.

[0050] The self-assembling peptides of the present disclosure may have nanofiber diameters ranging from about 10 nm to about 20 nm, with average pore sizes ranging from about 5 nm to about 200 nm. In certain embodiments, the nanofiber diameter, pore size, and nanofiber density may be controlled by at least one of the concentration of the peptide solution used and the amount of peptide solution used (e.g., volume of peptide solution). Thus, a specific concentration of peptide in solution and / or a specific amount of peptide solution may be selected to provide a desired nanofiber diameter, pore size, and / or density for proper delivery and embolization upon administration to a biological vessel.

[0051] As used herein, an "effective amount" or "therapeutically effective amount" refers to an amount of a peptide, peptide solution or hydrogel that, upon single or multiple administrations (applications or injections) to a subject, is more effective in treating, or in curing, alleviating, ameliorating or improving a subject with a disorder than would be expected in the absence of such treatment. This may include a particular concentration or concentration range of peptide in the peptide solution or hydrogel, or even, or alternatively, a particular volume or volume range of the peptide solution or hydrogel. A method for promoting may include providing instructions for preparing at least one of the effective amount and effective concentration.

[0052] The dosage administered (e.g., applied or injected), e.g., volume or concentration, may vary depending on the form of the peptide (e.g., in a peptide solution, in a hydrogel, or in a dry form (e.g., lyophilized form)) and the route of administration utilized. The exact formulation, route of administration, volume and concentration may be selected in view of the condition of the subject and in view of the particular target area or location to which the peptide solution, hydrogel, or other form of peptide is administered. Lower or higher doses than those listed herein may be used or may be required. The specific dosage and treatment regimen for any particular subject may depend on a variety of factors, which may include the specific peptide(s) used, the dimensions of the biological vessel to be treated or occluded, the desired thickness of the resulting hydrogel that may be placed in the desired target area, and the length of time of treatment. Other factors that may affect the specific dosage and treatment regimen include age, weight, general health, sex, administration time, degradation rate, severity and course of disease, condition or symptoms, and the judgment of the treating physician. In certain embodiments, the peptide solution may be administered in a single dose, hi other embodiments, the peptide solution may be administered in more than one dose, or in multiple doses.

[0053] An effective amount and effective concentration of the peptide solution may be selected to at least partially occlude or block a biological vessel. In some embodiments, at least one of the effective amount and effective concentration may be based in part on the diameter of a target region of the biological vessel. In other embodiments, at least one of the effective amount and effective concentration may be based in part on the flow rate of blood in the biological vessel. In other embodiments, at least one of the effective amount and effective concentration may be based in part on the blood pressure of blood in the biological vessel. In yet other embodiments, at least one of the effective amount and effective concentration may be based in part on the average diameter of the subject's red blood cells.

[0054] In still other embodiments, at least one of the effective amount and the effective concentration may be based in part on at least one of the diameter of the target region of the biological vessel, the flow rate of blood in the biological vessel, the blood pressure of blood in the biological vessel, and the average diameter of red blood cells of the subject.

[0055] At least one of the effective amount and the effective concentration may be based in part on providing hydrogel nanofibers having an average pore size smaller than the average diameter of the subject's red blood cells, which may include determining a mean red blood cell diameter and collecting a blood sample from the subject to provide at least one of the effective amount and the effective concentration.

[0056] The effective amount may be an amount that can provide a desired blockage in a biological vessel, as described herein. Various characteristics of the biological vessel may contribute to the selection or determination of the effective amount, including at least one of the diameter of a target region of the biological vessel, the flow rate of blood in the biological vessel, the blood pressure of blood in the biological vessel, and the average diameter of red blood cells of the subject.

[0057] The effective amount may comprise a volume of about 0.1 milliliters (mL) to about 100 mL of peptide solution. The effective amount may comprise a volume of about 0.1 mL to about 10 mL of peptide solution. In certain embodiments, the effective amount may be about 0.5 mL. In other embodiments, the effective amount may be about 1.0 mL. In still other embodiments, the effective amount may be about 1.5 mL. In still yet other embodiments, the effective amount may be about 2.0 mL. In some other embodiments, the effective amount may be about 3.0 mL.

[0058] In some embodiments, more effective blocking may be achieved with a larger volume of peptide solution administered. This may allow a longer or thicker hydrogel to form within the biological vessel, allowing for a more reliable location of the hydrogel in the target area. It is conceivable that if a high enough volume is not selected, the hydrogel may not be effective in maintaining blocking in the target area for a desired period of time. This may be affected by the blood flow rate or blood pressure in the vessel.

[0059] The effective concentration may be an amount that may provide a desired blockage in a biological vessel, as described herein. Various characteristics of the biological vessel may contribute to the selection or determination of the effective concentration, including at least one of the diameter of a target region of the biological vessel, the flow rate of blood in the biological vessel, the blood pressure of blood in the biological vessel, and the average diameter of red blood cells of the subject.

[0060] The effective concentration may include a peptide concentration in the solution ranging from about 0.1 weight / volume (w / v)% to about 10 w / v%. The effective concentration may include a peptide concentration in the solution ranging from about 0.1 w / v% to about 3.5 w / v%. In certain embodiments, the effective concentration may be about 1 w / v%. In other embodiments, the effective concentration may be about 2.5 w / v%. In yet other embodiments, the effective concentration may be about 3.0 w / v%.

[0061] In certain embodiments, a peptide solution with a higher concentration of peptide may provide a more effective hydrogel that has the ability to remain in place and provide effective blocking of the biological vessel. For the purpose of delivering the peptide solution, a higher concentration of peptide solution may become too viscous to allow effective and selective administration of the solution. It is conceivable that if a high enough concentration is not selected, the hydrogel may not be effective in maintaining blocking in the target area for a desired period of time. This may also be affected by the blood flow rate or blood pressure in the vessel.

[0062] The effective concentration can be selected to provide a solution that can be administered by injection or other means, using a catheter or needle of a particular diameter or gauge.

[0063] The disclosed method contemplates single and multiple administrations of therapeutically effective amounts of peptides, peptide solutions and hydrogels as described herein. Peptides as described herein may be administered at regular intervals depending on the nature, severity and extent of the subject's condition. In some embodiments, peptides, peptide solutions, or hydrogels may be administered in a single dose. In some embodiments, peptides, peptide solutions, or hydrogels as described herein are administered in multiple doses. In some embodiments, therapeutically effective amounts of peptides, peptide solutions, or hydrogels may be administered periodically at regular intervals. The regular intervals selected may be based on any one or more of the initial peptide concentration of the solution administered, the amount administered, and the degradation rate of the hydrogel formed. For example, after a first administration, a follow-on administration may be performed, for example, 2 weeks, 4 weeks, 6 weeks, or 8 weeks later. The follow-on administration may include administration of a solution having the same peptide concentration and volume as the first administration, or may include administration of a solution with a lower or greater concentration and volume of peptide. Selection of an appropriate next administration of peptide solution may be based on imaging the target area and the area around the target area to ascertain the need based on the subject's condition. The predetermined interval may be the same or different for each next administration. In some embodiments, peptides, peptide solutions, or hydrogels may be administered chronically at predetermined intervals to maintain at least partial blockage of biological vessels in the subject throughout the subject's life. The predetermined interval may be the same or different for each next administration. This may depend on whether the hydrogel formed from the previous administration has partially or completely collapsed or degraded. The next administration may include administration of a solution having the same concentration of peptide and volume as the first administration, or may include administration of a solution with a lower or higher concentration and volume of peptide.Selection of an appropriate subsequent administration of peptide solution may be based on imaging the target area and the area surrounding the target area to ascertain the need based on the subject's condition.

[0064] The self-assembling peptides of the present disclosure (e.g., RADA16 (SEQ ID NO: 7)) may be peptide sequences that lack distinct physiologically or biologically active motifs or sequences and therefore may not impair essential cellular functions. Physiologically active motifs may control many intracellular events (e.g., transcription), and the presence of the physiologically active motif may result in phosphorylation of cytoplasmic or cell surface proteins by enzymes that recognize the motif. When a physiologically active motif is present in a peptide tissue-blocking agent, the transcription of proteins with various functions may be activated or suppressed. The self-assembling peptides of the present disclosure do not have this risk, since they may lack such physiologically active motifs.

[0065] Sugars can be added to the self-assembling peptide solution to improve the osmolality of the solution from hypotonic to isotonic without reducing tissue occlusion effects, thereby allowing for increased biological safety. In certain instances, the sugar can be sucrose or glucose.

[0066] In certain embodiments, the length of the peptide can be longer than 12 amino acids, preferably at least 16 residues.Very long peptides, for example peptides of about 200 amino acids, may encounter problems due to insolubility and intramolecular interactions that destabilize membrane formation, but can also be contemplated herein.In addition, peptides with a large amount of hydrophobic residues may have insolubility problems.The optimal length for membrane formation may vary depending on amino acid composition.

[0067] An additional stabilizing factor is that complementary peptides maintain a certain distance between the peptide backbones. Peptides that can maintain a certain distance during pairing are said to be structurally compatible herein. Interpeptide distance can be calculated for each ionized or hydrogen-bonded pair by summing the number of unbranched atoms on the side chain of each amino acid in the pair. For example, lysine has five and glutamic acid has four unbranched atoms on its side chain, respectively.

[0068] Examples of peptides capable of forming membranes in a homogenous mixture are shown in Table 1. These examples illustrate some of the various amino acid arrangements and compositions of membrane-forming peptides.

[0069] [Table 1-1]

[0070] [Table 1-2]

[0071] The criteria of amphipathic sequence, length, complementarity and structural compatibility apply to a heterogeneous mixture of peptides.For example, two different peptides can be used to form a membrane: peptide A, Val-Arg-Val-Arg-Val-Asp-Val-Asp-Val-Arg-Val-Arg-Val-Asp-Val-Asp (VRVRVDVDVRVRVDVD) (SEQ ID NO: 54), has Arg and Asp as hydrophilic residues, as shown in the attached sequence listing, and peptide B, Ala-Asp-Ala-Asp-Ala-Lys-Ala-Lys-Ala-Asp-Ala-Asp-Ala-Lys-Ala-Lys (ADADAKAKADADAKAK) (SEQ ID NO: 55), has Lys and Asp. Peptides A and B are complementary; Arg on A can form an ionized pair with Asp on B, and Asp on A can form an ionized pair with Lys on B. However, calculation of the distance between the peptides in such pairs shows that the two peptides are structurally incompatible. Using a conversion factor of 3 Å per atom, the difference in interpeptide distance between the two pairs is 3 Å. Variations in interpeptide distance beyond 3-4 Å are predicted to destabilize the intermolecular interactions, leading to membrane formation. Thus, in heterogeneous mixtures of peptides A and B, membranes are likely to form, but they are composed uniformly of either peptides A or B.

[0072] Membranes and hydrogels can also be formed from heterogeneous mixtures of peptides (each of which would not form a membrane by itself) if the peptides are complementary and structurally compatible with one another. For example, (Lys-Ala-Lys-Ala). 4 (KAKA) 4 (SEQ ID NO: 12) and (Glu-Ala-Glu-Ala) 4 (EAEA) 4 (SEQ ID NO: 56) or (Lys-Ala-Lys-Ala) 4 (KAKA) 4 (SEQ ID NO: 12) and (Ala-Asp-Ala-Asp) 4 (ADAD) 4(SEQ ID NO:57) mixture is predicted to form membranes, but not any of these peptides alone due to lack of complementarity.

[0073] Peptides may be considered to be neither completely complementary nor structurally compatible, but to contain mismatches similar to the mismatched base pairs in nucleic acid hybridization. Peptides containing mismatches may form membranes if the disruptive force of the mismatched pair is dominated by the overall stability of the peptide-peptide interaction. Functionally, such peptides may also be considered to be complementary or structurally compatible. For example, a mismatched amino acid pair may be tolerated if it is surrounded by several perfectly matched pairs on each side. Mismatched peptides may be tested for their ability to self-assemble into macroscopic membranes using the methods described herein.

[0074] The peptides may be chemically synthesized or purified from natural and recombinant sources. Using chemically synthesized peptides can ensure that the peptide solution is devoid of unidentified components (e.g., unidentified components derived from the extracellular matrix of another animal). This characteristic can therefore eliminate infection concerns (including the risk of viral infection) compared to traditional tissue-derived biomaterials. This can eliminate concerns such as infectious diseases, including infectious diseases such as bovine spongiform encephalopathy (BSE), making the peptides very safe for medical use.

[0075] The initial peptide concentration may be a factor in the size and thickness of the film or hydrogel formed. In general, the higher the peptide concentration, the greater the degree of film formation. Films or hydrogels may form from initial peptide concentrations as low as about 0.5 mM or about 1 mg / ml (about 0.1 w / v %). However, films or hydrogels formed with higher initial peptide concentrations (about 10 mg / ml (about 1 w / v %)) may be thicker and therefore perhaps stronger. When producing the film or hydrogel, it may be preferable to add peptide to a salt solution or physiological conditions rather than adding salt to the peptide solution.

[0076] The formation of the film or hydrogel can be very fast, on the order of a few minutes. The formation of the film or hydrogel can be immediate upon application or injection to the desired area. The formation of the film or hydrogel can occur within 1-2 minutes of application or injection. In another example, the formation of the film or hydrogel can occur within 4 minutes of application or injection. In certain embodiments, the time it takes to form the film or hydrogel can be based at least in part on one or more of the concentration of the peptide solution, the volume of the peptide solution applied, and the conditions at the area of ​​application or injection (e.g., the concentration of monovalent metal cations at the area of ​​application, blood flow rate, blood pressure, and the diameter of the biological vessel).

[0077] The formation of the film or hydrogel may be irreversible. The process may not be affected by pH below 12 (the peptides tend to precipitate at pH above 12) and by temperature. The film or hydrogel may form at temperatures ranging from 4 to 90°C.

[0078] The membrane or hydrogel can remain in place in the target area for a sufficient period of time to provide a desired effect using the disclosed method and kit. The desired effect can be to reduce or impede the flow of fluid through a biological pathway or channel. The desired effect can be a blockage, retention, obstruction, or embolization in one or more biological pathways or channels. The desired effect can be to intentionally create such a blockage, retention, or obstruction to remove (perfuse) a tumor or other pathological process from its blood supply.

[0079] The desired effect of using the disclosed method and kit can be to treat a disorder, malformation, or congenital disease in biological vessels. The desired effect can be to treat one or more of patent ductus arteriosus (PDA), major aortopulmonary collateral artery (MAPCA), recurrent hemoptysis, arteriovenous malformation, cerebral aneurysm, gastrointestinal bleeding, epistaxis, postpartum bleeding, intraoperative bleeding, and uterine fibroids. The desired effect can include providing at least partial blockage to cause cell necrosis or reduce or eliminate cancer cells.

[0080] The period during which the membrane or hydrogel may remain in the desired area may be about 10 minutes. In certain instances, it may remain in the desired area for about 35 minutes. In certain further instances, it may remain in the desired area for several days, up to two weeks. In other instances, it may remain in the desired area indefinitely. In other instances, it may remain in the desired area for a longer period of time until it is naturally degraded or intentionally removed. When the hydrogel is naturally degraded over a period of time, subsequent application or injection of the hydrogel into the same or different location in the biological vessel, or into another biological vessel, may be performed.

[0081] In certain embodiments, the self-assembling peptide may be prepared with one or more components that may provide enhanced effectiveness of the self-assembling peptide, provide another action, treatment, therapy, or interact with one or more components of the subject in another way. For example, an additional peptide that includes one or more biologically or physiologically active amino acid sequences or motifs may be included as one of the components with the self-assembling peptide. Other components may include biologically active compounds (e.g., drugs or other treatments that may provide some benefit to the subject). For example, drugs or anti-cancer drugs that treat cancer may be administered together with the self-assembling peptide or may be administered separately.

[0082] The peptide, peptide solution, or hydrogel may contain a small molecule drug for treating the subject or for preventing hemolysis, inflammation, and infection. The small molecule drug may be selected from the group consisting of glucose, saccharose, purified saccharose, lactose, maltose, trehalose, dextran, iodine, lysozyme chloride, dimethylisopropylazulene, tretinoin tocopheryl, povidone iodine, alprostadil alfadex, anise alcohol, isoamyl salicylate, α,α-dimethylphenylethyl alcohol, bacdanol, helional, sulfazin silver, bucladesine sodium, alprostadil alfadex, gentamicin sulfate, tetracycline hydrochloride, sodium fusidate, mupirocin calcium hydrate, and isoamyl benzoate. Other small molecule drugs may also be contemplated. Protein-based drugs may be included as administered components and these may include erythropoietin, tissue plasminogen activator, synthetic hemoglobin and insulin.

[0083] Components may be included to protect the peptide solution from rapid or immediate formation into a hydrogel. This may include an encapsulation delivery system that may degrade over time to allow for time-controlled release of the peptide solution to a target area to form the hydrogel over a desired, predetermined period of time. Biodegradable, biocompatible polymers may be used (e.g., ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid).

[0084] Any of the components described herein may be included in the peptide solution or may be administered separately from the peptide solution. Additionally, any of the methods or methods for promoting provided herein may be performed by one or more participants.

[0085] The peptide, peptide solution, or hydrogel of the present disclosure may be provided in a kit. Instructions for administering the solution to a biological vessel in a subject may also be provided in the kit. The peptide solution may include an amphipathic peptide that includes at least 12 amino acids, alternating hydrophobic and hydrophilic amino acids, in an amount and at an effective concentration to form a hydrogel under physiological conditions to allow at least partial blocking of a biological vessel. Instructions for administering the solution may include a method for administering the peptide, peptide solution, or hydrogel provided herein at a dose, volume, or concentration, or administration schedule, for example, by an administration route described herein.

[0086] The kit may also include informational materials. The informational materials may be explanatory, instructional, marketing or other materials regarding the methods described herein. In one embodiment, the informational materials may include information regarding the manufacture of the peptides, peptide solutions or hydrogels disclosed herein, physical properties, concentrations, volumes, sizes, dimensions, expiration dates, and batches or production locations of the peptides, peptide solutions or hydrogels.

[0087] The kit may also include devices or materials as necessary to allow administration of the peptide or peptide solution to a desired area. For example, a syringe, pipette, catheter, or other needle-based device may be included in the kit. Additionally or alternatively, the kit may include a guidewire or other associated equipment to provide selective administration of the peptide solution to a target area.

[0088] The kit may further or alternatively include other components or ingredients, such as components that may assist in contrast imaging. For example, the kit may include a contrast agent. The contrast agent may provide a visual image during the use of X-ray techniques, such as fluoroscopy or angiography. A non-ionic radiopaque contrast medium may be included, such as a water-soluble iodine-based solution. The water-soluble iodine-based solution may be iopamidol. Instructions may be provided in the kit to combine a sufficient amount or volume of the contrast agent to properly at least one of: identify the target area, introduce a catheter or other administration device, place the end of the catheter in the target area, administer the peptide solution, remove the catheter or other administration device, and observe the biological vessel after removing the catheter. The use of the contrast agent may allow visualization of the area where the peptide, peptide solution, or hydrogel is administered. Instructions may be provided to dilute the peptide solution to administer an effective concentration of the solution to the biological vessel. Instructions may further be provided for determining at least one of an effective concentration of the solution and an effective amount of the solution for the biological vessel, which may be based on various parameters discussed herein, including the diameter of the biological vessel in the target area.

[0089] Other components or ingredients may be included in the kit in the same or different compositions or containers other than the peptide, peptide solution, or hydrogel. The one or more components may include components that can provide enhanced effectiveness of the self-assembling peptide, provide another action, treatment, therapy, or interact with one or more of the subject components in another way. For example, an additional peptide containing one or more biologically or physiologically active sequences or motifs may be included as one of the components with the self-assembling peptide. Other components may include biologically active compounds, such as drugs or other treatments that can provide some benefit to the subject. For example, cancer treatment drugs or anti-cancer drugs may be administered together with the self-assembling peptide or may be administered separately. The peptide, peptide solution, or hydrogel may include small molecule drugs to treat the subject or prevent hemolysis, inflammation, and infection as disclosed herein. A sugar solution (e.g., a sucrose solution) may be provided in the kit. The sucrose solution may be a 20% sucrose solution.

[0090] Other components disclosed herein may also be included in the kit.

[0091] In some embodiments, the kit components are stored in sealed vials (e.g., with rubber or silicone closures, such as polybutadiene or polyisoprene closures). In some embodiments, the kit components are stored under inert conditions (e.g., under nitrogen or another inert gas, such as argon). In some embodiments, the kit components are stored under anhydrous conditions (e.g., with a desiccant). In some embodiments, the kit components are stored in light-proof containers (e.g., amber vials).

[0092] As part of the kit or separately from the kit, the syringe or pipette can be pre-filled with peptide, peptide solution, or hydrogel as disclosed herein.Methods are provided to instruct the user to supply the self-assembling peptide solution to the syringe or pipette with or without other device, and administer the solution to the target area through the syringe or pipette with or without other device.Other devices can include, for example, catheters with or without guidewires.

[0093] In some embodiments of the present disclosure, the self-assembling peptides can be used as coatings on devices or instruments, such as stents or catheters, to suppress fluid leakage.The self-assembling peptides can also be incorporated into or fixed to supports (e.g., gauze or bandages) or linings that can provide therapeutic effects to subjects or be applied within biological vessels.The self-assembling peptides can also be impregnated into sponges for use.

[0094] In an alternative embodiment, an atomizing sprayer can be prepared that is filled with a powder or solution of the self-assembling peptides. When such a sprayer is used to spray an affected area, its pH and salt concentration increases upon contact with the body, causing gelation.

[0095] The modification of the membrane can give it additional properties. For example, the membrane can be further strengthened by crosslinking the peptide after membrane formation by standard methods. Collagen can be combined with the peptide to produce a membrane more suitable for use in artificial skin; the collagen can be stabilized from proteolysis within the membrane. Furthermore, phospholipids can be combined with the peptide to produce vesicles.

[0096] The membrane may also be useful for culturing cell monolayers. Cells prefer to adhere to heterogeneously charged surfaces. The charged residues and conformation of the proteinaceous membrane promote cell adhesion and migration. The addition of growth factors (e.g., fibroblast growth factor) to the peptide membrane may further improve adhesion, cell proliferation and neurite outgrowth.

[0097] The function and advantages of these and other embodiments of the methods and kits disclosed herein will be more fully understood from the following examples, which are intended to illustrate the benefits of the disclosed treatment approaches, but do not exemplify their full scope. EXAMPLES

[0098] Example 1 3% (weight / volume (w / v)) PuraMatrix TM Solution, Ac-RADARADARADARADA-NH 2 (Ac-Arg-Ala-Asp-Ala-Arg-Ala-Asp-Ala-Arg-Ala-Asp-Ala-Arg-Ala-Asp-Ala-NH 2 A study was performed in rats using a peptide solution containing (SEQ ID NO: 58) in water. One milliliter (mL) was injected into the portal vein of the rat using an 18 gauge needle.

[0099] Histopathological evaluation was performed using hematoxylin-eosin (HE) dye. It was confirmed that embolization occurred in the portal vein using the peptide solution. As shown in Figure 1, the peptide solution appears to have developed into a hydrogel 2 and is present in the portal vein of the rat. Red blood cells 4 are also shown.

[0100] Example 2 2.5% PuraMatrix TM Solution, Ac-RADARADARADARADA-NH 2(Ac-Arg-Ala-Asp-Ala-Arg-Ala-Asp-Ala-Arg-Ala-Asp-Ala-Arg-Ala-Asp-Ala-NH 2 Studies were conducted in two beagle dogs using a peptide solution containing 2.5% peptide (SEQ ID NO: 58) in water. These studies confirmed the efficacy of hepatic artery embolization and hepatocellular necrosis in beagle dogs using the 2.5% peptide solution. The peptide solution contained iopamidol at a concentration of 612.4 mg / mL. Iopamidol is a non-ionic radiopaque contrast agent.

[0101] Under full anesthesia, a beagle dog was X-ray imaged, and a microcatheter (Terumo, minimum inner diameter 0.50 mm) was inserted into the hepatic artery via the carotid artery. Hepatic arteriography imaging was used to confirm that the hepatic artery was functional. A 2 mL volume of the above 2.5% peptide solution (containing iopamidol) was injected.

[0102] Hepatic arterial contrast imaging was used to confirm that embolization of the hepatic artery with peptide solution was effected during surgery. Figure 2 shows a normal hepatic arterial contrast image of a beagle dog, while Figure 3 shows the peptide solution injection. Figure 4 shows a hepatic arterial contrast image after peptide solution injection, where back flow of the contrast image was confirmed. The presence of the peptide solution is revealed by the darker areas of the image.

[0103] After 2 weeks of monitoring, hepatic arteriogram imaging was used to confirm the embolization effect. Figure 5 shows a hepatic arteriogram 2 weeks after injection of the peptide solution.

[0104] The liver was then removed. Hematoxylin-eosin (HE) dye was used to histopathologically confirm the peptide solution embolism and liver damage. As shown in Figures 6A-6C, the peptide solution can be seen in each of these images of the hepatic artery as a dark area of ​​the image. Figure 7 shows a hepatocyte necrosis image at the embolization location, where all cells appear to have at least some level of necrosis.

[0105] The results show that injection of the peptide solution using a microcatheter can be achieved. The peptide gel can be revealed using X-ray imaging. Hepatic artery embolization effects can be seen during surgery and two weeks after surgery. Furthermore, hepatic artery embolization effects and hepatocyte necrosis effects using the peptide solution occurred and were confirmed histopathologically.

[0106] Example 3 PuraMatrix TM , Ac-RADARADARADARADA-NH 2 (Ac-Arg-Ala-Asp-Ala-Arg-Ala-Asp-Ala-Arg-Ala-Asp-Ala-Arg-Ala-Asp-Ala-NH 2A peptide solution containing 5'-diaminobutyric acid (SEQ ID NO: 58) in water was used as an embolic formulation in a porcine model through angiography, gross necropsy assessment, and histopathology evaluation. One female Yorkshire cross swine was studied. The pig weighed 46.5 kg at the time of study. Food and water were provided according to standard operating procedures. There was no contamination of the food or water that would be expected to interfere with the management or outcome of the study. The pig was obtained from a testing facility approved animal supplier. The pig was physically examined upon arrival and examined again after an acclimation period. The pig was fasted for a minimum of 12 hours prior to the procedure. The animals were sedated and anesthetized with intramuscular or subcutaneous injections of Telazol (2-10 mg / kg) and Xylazine (0.5-5.0 mg / kg). Propofol (until effective) was given to aid in sedation. An endotracheal tube was used to ensure adequate ventilation and the animals were maintained under general anesthesia with inhalant isofluorante (0.1-5.0%). Heparin (50-300 units / kg, IV) was administered throughout the procedure.

[0107] A 2.5% test solution of the peptide solution was used. Approximately 800 μl of the peptide solution was placed in an Eppendorf tube. Approximately 200 μl of Isovue-370 (iopamidol) contrast agent was added. The liquid was mixed slowly to avoid air bubbles.

[0108] On the day of the study, the pig was 2 months and 25 days old. The pig was sedated and prepared for surgery. The femoral artery was accessed and an introducer was placed. A guidewire was advanced into the selected renal artery. A catheter was advanced into the selected renal artery. Angiography was used to visualize the location within the artery. The peptide solution was injected into the desired location until the artery was occluded. This procedure was repeated in the hepatic and splenic arteries. Angiography was used throughout the procedure to visualize the vessels and devices throughout the study. Figures 8 and 9 are representative examples of the vessels before (Figure 6) and after (Figure 9) embolization. There were no adverse events reported throughout the study.

[0109] A summary of the data can be found in Table 2 below.

[0110] [Table 2]

[0111] As shown in Table 2, injection into the renal artery of the left kidney was successful immediately after injection. Successful embolization of the renal artery of the right kidney was successful immediately after injection, although slight flow was re-established 6 minutes after the first injection. An additional injection was performed 15 minutes after the first injection, resulting in complete occlusion.

[0112] Successful embolization of the hepatic artery was also obtained immediately after injection, with the remaining artery being occluded after 10 and 34 minutes.

[0113] Successful embolization of the splenic artery was also obtained after 4 minutes.

[0114] The description and drawings provided are illustrative only and are not to be construed as limiting. While exemplary embodiments of the present disclosure have been disclosed, many modifications, additions, and deletions may be made therein without departing from the spirit and scope of the disclosure and its equivalents, as set forth in the following claims.

[0115] Those skilled in the art will readily recognize that the various configurations described herein are meant to be exemplary, and that the actual configuration will depend on the specific application for which the disclosed systems and methods are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments described herein.

[0116] Furthermore, it should be recognized that various changes, modifications, and improvements will readily occur to those skilled in the art. Such changes, modifications, and improvements are to be considered part of this disclosure and are to be considered within the spirit and scope of this disclosure. Accordingly, the foregoing description and drawings are for illustrative purposes only. Furthermore, the depictions in the drawings do not limit the disclosure to the representations specifically shown.

[0117] As used herein, the terms "comprising," "including," "carrying," "having," "containing," and "involving," whether in the written specification, the claims, or otherwise, are open-ended terms, i.e., mean "including, but not limited to." Thus, use of such terms is meant to encompass the items listed thereafter, and equivalents thereof, as well as additional items. Only the traditional phrases "consisting of" and "consisting essentially of" are closed or semi-closed traditional phrases, respectively, with respect to the claims. The use of ordinary terms in the claims, such as "first," "second," "third," etc., to modify claim elements does not, by itself, imply any priority, precedence, or ordering of one claim element over another, or the chronological order in which acts of a method are performed, but is merely used as a label to distinguish one claim element having a certain name from another element having the same name (in the absence of ordinary use of the term).

Claims

[Claim 1] A composition or method as described in the specification.

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