Transdermal delivery device and method of use for peptide delivery

Abstract

Patches, methods, devices, and systems for delivering non-aggregating peptides such as arcadien and fragments thereof are disclosed herein. In some embodiments, the patch includes a backing, a matrix containing a non-aggregating peptide disposed within the matrix, and a release liner. In other embodiments, the method includes opening at least one channel in the skin of a subject and applying a patch as described herein, thereby treating a brain-related disease or disorder such as Alzheimer's disease.

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 350,982, filed on June 10, 2022, which is incorporated herein by reference in its entirety.

[0002] Reference to Sequence Listing This application is filed with an electronic sequence listing. The sequence listing is provided as a file entitled PPORT.008WO_ST_26.xml, created on June 6, 2023, with a size of approximately 7 kb. The electronic sequence listing information is incorporated herein by reference in its entirety.

[0003] Background Field This application relates to compositions, devices, and methods for transdermal drug delivery, and particularly to peptide compositions and methods for administering peptides to a subject by a transdermal microporation device.

[0004] Background Art Alzheimer's disease (AD) is the most common neurodegenerative disorder. The increasing number of AD patients is a serious global problem. Although familial genetic mutations that increase the risk of developing AD have been identified, most of these cases are thought to be due to age - related cognitive decline in the brain. Therefore, aging is the greatest risk factor for developing AD, but a characteristic change commonly seen in AD patients is the accumulation of amyloid - β (Aβ) in the brain. Aβ, which increases with age, forms aggregates of Aβ oligomers in the brain and develops cytotoxicity. Since cognitive function decline caused by Aβ oligomers has been reported, Aβ is considered a strong cause of AD onset.

[0005] Alcadein β, a neuron-specific membrane protein, is metabolized through a pathway similar to that of Aβ to produce p3-Alcβ. Unlike Aβ, which exponentially accumulates in the brain with aging, p3-Alcβ does not accumulate in the brain but decreases in cerebrospinal fluid (CSF) with aging, and p3-Alcβ in CSF is further significantly decreased in AD patients compared to age-matched non-dementia subjects, suggesting its involvement in the onset of AD. Furthermore, since p3-Alcβ does not have aggregability unlike Aβ, it reduces the neurotoxicity and cognitive function caused by Aβo by peripheral administration in an acute cognitive function model mouse induced by Aβ oligomers. It has been clarified that this shows a reducing and improving effect. Therefore, it is considered that an increase in toxic Aβ oligomers and a decrease in p3-Alcβ having an inhibitory effect are involved in the onset and progression of AD. Since the action of p3-Alcβ is carried out by its partial peptide p3-Alcβ9-19, the onset and progression of AD associated with the decrease in the endogenous molecule p3-Alcβ can be prevented by reaching p3-Alcβ9-19 to the brain.

[0006] Various peptide compositions and treatment methods have been suggested for AD. However, peptide drugs are generally unstable in vivo and difficult to deliver to the brain. Therefore, there remains a long-standing need for improved peptides, formulations, compositions, devices, and methods for the delivery of peptides across the blood-brain barrier.

[0007] Summary of the Invention Some aspects of the present disclosure relate to patches for delivering non-aggregated peptides. In some embodiments, the patch includes a backing, a matrix containing the non-aggregated peptide disposed within the matrix, and a release liner configured to be removed prior to application to the skin of a subject. In some embodiments, the non-aggregated peptide is p3-Alcβ. In some embodiments, p3-Alcβ is selected from at least one of p3-Alcβ1-40, p3-Alcβ1-37, p3-Alcβ9-19, p3-Alcβ1-19, p3-Alcβ11-19, or a derivative thereof. In some embodiments, p3-Alcβ is p3-Alcβ9-19 or a derivative thereof. In some embodiments, the non-aggregated peptide is about 0.01 mg / cm 2 ~200 mg / cm 2is an amount within the range. In some embodiments, the matrix further comprises at least one sugar. In some embodiments, the at least one sugar is selected from non-reducing sugars, reducing sugars, or combinations thereof. In some embodiments, the non-reducing sugar is selected from sucrose, trehalose, mannitol, sorbitol, or combinations thereof. In some embodiments, the reducing sugar is selected from lactose, maltose, or combinations thereof. In some embodiments, the ratio of the at least one sugar to the non-aggregating peptide is greater than 0.02. In some embodiments, the ratio of the at least one sugar to the non-aggregating peptide is from about 0.02 to about 0.4. In some embodiments, the matrix further comprises an organic acid, an organic salt, or combinations thereof. In some embodiments, the organic acid is a pharmacokinetic (PK) modulator. In some embodiments, the PK modulator is citric acid and its salt forms. In some embodiments, the matrix further comprises a preservative. In some embodiments, the preservative is an antimicrobial agent. In some embodiments, the antimicrobial agent is selected from methylparaben, propylparaben, benzalkonium chloride, and sodium benzoate. In some embodiments, the matrix comprises at least one fiber. In some embodiments, the at least one fiber is a non-woven fiber. In some embodiments, the at least one fiber has a thickness of less than 300 μm. In some embodiments, the at least one fiber has a weight of less than 100 g / m 2 and has a weight of less than. In some embodiments, the matrix has a water holding capacity of less than 10 mg / cm 2 . In some embodiments, the matrix comprises a film laminating material and at least one fiber. In some embodiments, the matrix further comprises at least one of sucrose, lactose, disodium citrate sesquihydrate, methylparaben, propylparaben, and benzalkonium chloride.

[0008] One aspect relates to a method of treating a disease or condition associated with a subject's brain. In some embodiments, the method includes opening at least one channel in the subject's skin and applying the patch described herein to the subject's skin, thereby treating a disease or condition associated with the brain. In some embodiments, the disease or condition associated with the brain is a neurodegenerative disease. In some embodiments, the neurodegenerative disease is Alzheimer's disease. In some embodiments, opening at least one channel in the subject's skin includes applying a transdermal microporation device to the subject's skin. In some embodiments, the transdermal microporation device utilizes thermal tissue ablation by using a filament array having a plurality of filaments disposed on the subject's skin, and each filament can conductively deliver thermal energy through direct contact with the tissue membrane to form a plurality of micropores in the micropore region of the tissue membrane. In some embodiments, the transdermal microporation device generates from about 25 to about 500 microchannels / cm 2 In some embodiments, the transdermal microporation device has a poration energy of from about 2 to about 6 mJ / filament. In some embodiments, opening at least one channel in the subject's skin has an area of from about 0.25 cm 2 to about 4 cm 2 . In some embodiments, the transdermal microporation device is a micro-needle, laser, or radiofrequency device capable of generating one or more micropores in the subject's skin. In some embodiments, applying the patch increases the neuron survival rate in the subject. In some embodiments, applying the patch increases the mitochondrial activity in the subject's brain. In some embodiments, the patch provides a maximum blood concentration of the non-aggregated peptide at least 0.5 hours after administration. In some embodiments, the non-aggregated peptide is maintained for at least 6 hours after administration of the patch. In some embodiments, the non-aggregated peptide from the patch is maintained in the subject's blood for at least 6 hours after administering the patch to the subject. In some embodiments, the transfer rate of the non-aggregated peptide from the subject's blood to the subject's cerebrospinal fluid is at least 2%.

[0009] One aspect relates to a transdermal delivery system for delivering a non-aggregated peptide. In some embodiments, the system comprises a patch as described herein and a transdermal microporation device. In some embodiments, the transdermal microporation device includes an applicator electrically connected to an array of conductive filaments, and the transdermal microporation device is configured to generate thermal energy based on a current flowing through the array of conductive filaments and supply the thermal energy to a biological membrane disposed adjacent to the transdermal microporation device, and a power supply circuit is configured to supply a current to the transdermal microporation device. In some embodiments, the applicator supplies a predetermined electrical energy to the array of conductive filaments, thereby generating one or more micropores. In some embodiments, the patch is applied to the one or more micropores. In some embodiments, the transdermal microporation device generates about 25 to about 500 microchannels / cm 2 In some embodiments, the transdermal microporation device has a poration energy of about 2 to about 6 mJ / filament. In some embodiments, the step of opening at least one channel in the subject's skin has an area of about 0.25 cm 2 to about 4 cm 2 In some embodiments, one or more micropores are about 0.5 to about 12.5% of the total poration area. In some embodiments, the transdermal microporation device is a microneedle, laser, or radiofrequency device capable of generating one or more micropores in the subject's skin. In some embodiments, the transdermal microporation device generates at least 50 pores in the subject's skin. In some embodiments, the patch further comprises a drug pellet.

[0010] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate several aspects described below and, together with the description, serve to explain the principles of the disclosure. Like numbers represent like elements throughout the drawings.

Brief Description of the Drawings

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[0012] Modes for Carrying Out the Invention This disclosure can be more easily understood by referring to the following detailed description, examples, drawings, and claims, as well as the descriptions before and after them. However, it should be understood that before the device, system, and / or method are disclosed and described, this disclosure is not limited to the specific devices, systems, and / or methods disclosed, unless otherwise specified. It should also be understood that the terms used herein are for the purpose of describing particular aspects only and are not necessarily intended to be limiting.

[0013] This description is provided as a teaching enabling the present disclosure. For this purpose, those skilled in the art will recognize and understand that many changes can be made to various aspects of the disclosure described herein while still obtaining beneficial results. It will also be apparent that some of the desired benefits can be obtained by selecting some of the features described herein without using other features. Thus, those skilled in the art will recognize that many modifications and adaptations to this description are possible and may even be desirable in certain circumstances and are part of the present disclosure. Accordingly, this description is provided as an illustration of certain principles of the present disclosure and is not intended to limit the present disclosure.

[0014] Definitions Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. All patents, applications, published applications, and other publications referenced herein are incorporated by reference in their entirety unless otherwise indicated. In the event of multiple definitions for terms herein, the definitions in this section shall control unless otherwise indicated.

[0015] The singular forms "a", "an", and "the" used throughout are to be construed to include the plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a filament" can include two or more such filaments unless the context clearly dictates otherwise.

[0016] Ranges can be expressed herein as from "about" a particular value and / or to "about" another particular value. When such a range is expressed, another aspect includes from a particular value and / or to another particular value. Similarly, when values are expressed using the antecedent "about" as approximations, it will be understood that the particular value forms another aspect. It will further be understood that the endpoints of each range are significant both in relation to the other endpoint and independently of the other endpoint.

[0017] As used herein, the terms "optional" or "optionally" mean that the subsequently described event or circumstance may or may not occur, and that the description includes both the case where the event or circumstance occurs and the case where it does not occur.

[0018] As used herein, "stratum corneum" refers to the outermost layer of the skin consisting of about 15 to about 20 layers of cells at various stages of drying. The stratum corneum provides a barrier against water loss from the body to the external environment and against attacks from the external environment to the body.

[0019] As used herein, "tissue" refers to an aggregate of specific types of cells that form a structural material, together with their intercellular substances. In the context of drug delivery to or through such tissue, at least one surface of the tissue is accessible to a transdermal delivery mode (e.g., a poration device and / or a patch). The tissue is the skin for the various poration delivery modes described herein. Other tissues suitable for use in the present disclosure include mucosal tissue and soft organs.

[0020] As used herein, the term "interstitial fluid" is a clear fluid that occupies the spaces between cells in the body.

[0021] As used herein, the term "biological fluid" is defined as a liquid derived from a biological organism, including serum or whole blood as well as interstitial fluid.

[0022] As used herein, "tissue membrane" can be any one or more epidermal layers of a subject. For example, in one embodiment, the tissue membrane is a skin layer that includes the outermost layer of the skin, i.e., the stratum corneum. In an alternative embodiment, the skin layer can include one or more backing layers of the epidermis commonly identified as the granular layer, the Malpighian layer, and the germinative layer. Those skilled in the art will understand that there is essentially little or no resistance to the transport or absorption of permeates through the backing layers of the epidermis. Thus, in one embodiment, at least one formed pathway within the skin layer of the subject is a pathway within the stratum corneum of the subject. Further, as used herein, "stratum corneum" typically refers to the outermost layer of the skin that contains approximately 15 to approximately 20 layers of cells at various stages of drying. The stratum corneum provides a barrier against water loss from the body to the external environment and against attacks from the external environment to the body. Further, as used herein, "tissue membrane" can refer to an aggregate of specific types of cells that form a structural material together with their intercellular substances. In various embodiments, at least one surface of the tissue membrane is accessible to one or more of the poration devices and / or permeate compositions described herein. As described above, the tissue membrane for various poration delivery modes is the skin. Other tissues suitable for use with such devices and compositions include mucosal tissues and soft organs.

[0023] As used herein, the term "subcutaneous fluid" can include, but is not limited to, water, plasma, blood, one or more proteins, interstitial fluid, and any combination thereof. In one embodiment, the subcutaneous fluid according to this description is a water source containing water.

[0024] As used herein, "poration", "microporation", or any such similar term means the formation of small holes or gaps (hereinafter also referred to as "micropores") in or through a tissue or biological membrane such as skin or mucosa, or the outer layer of an organism, for the purpose of reducing the barrier properties of the biological membrane to allow at least one permeate to pass from one side of the biological membrane to the other side for a selected purpose. Preferably, the holes or "micropores" so formed have a diameter of approximately 1 to 1000 microns and extend sufficiently into the biological membrane to disrupt the barrier properties of the stratum corneum without adversely affecting the underlying tissue. The term "micropore" is used in the singular for simplicity, but it should be understood that the microporation devices described herein can form multiple artificial openings. Poration can reduce the barrier properties of a biological membrane for a selected purpose or for a particular medical or surgical procedure. For the purposes of this application, "poration" and "microporation" are used interchangeably and mean the same thing.

[0025] A "microporator" or "porator" is a component of a microporation device capable of microporation. Examples of a microporator or porator include, but are not limited to, filaments that can conductively deliver thermal energy through direct contact with a biological membrane to cause ablation of a portion of the membrane deep enough to form micropores, optically heated local dye / absorber layers, electromechanical actuators, microlancets, micro needles or arrays of lancets, sonic energy ablators, laser ablation systems, high-pressure fluid jet punctures, etc. As used herein, "microporator" and "porator" are used interchangeably.

[0026] As used herein, "permeabilization" means the controlled removal of cells caused by the thermal energy and kinetic energy released when an explosive element detonates, whereby the cells of the biological membrane and, in some cases, some adjacent cells are "blown out" from that site. As used herein, "fusible" and "fusing" refer to an element that can remove itself from an electrical circuit when a sufficient amount of energy or heat is applied. That is, if a resistive electrically activated poration element is designed to be a fusible element, this means that when activated, during or after the formation of micropores in the biological membrane, the element is destroyed and the flow of current through it stops.

[0027] As used herein, "permeation enhancement" or "osmotic enhancement" means an increase in the permeability of a biological membrane and / or tissue to a drug, bioactive composition, or other chemical molecule, compound, or particle (also referred to as a "permeant") such that the rate at which the drug, bioactive composition, or other chemical molecule, compound, particle, or substance penetrates the biological membrane and / or tissue is increased.

[0028] As used herein, "enhancer", "chemical enhancer", "permeation enhancer", "osmotic enhancer", etc. include all enhancers that increase the flow of a permeant, analyte, or other molecule across a biological membrane, limited only by functionality. In other words, all cell envelope-disrupting compounds and solvents, as well as any other chemical promoters, are intended to be included. Furthermore, all active power enhancer techniques such as the application of sonic energy, mechanical suction, pressure, or local deformation of tissue, iontophoresis, or electroporation are included. One or more enhancer techniques can be combined sequentially or simultaneously. For example, a chemical enhancer can be applied first to permeabilize the capillary wall, and then an iontophoresis or sonic energy field can be applied to surround the capillary bed and actively push the permeant into the tissue containing the capillary bed.

[0029] As used herein, "transdermal" or "transdermally" means the passage of a permeant into and through a biological membrane to achieve an effective therapeutic blood level or local tissue level of the permeant, or the passage of an analyte molecule or liquid present in the body (an "analyte") through a biological membrane such that the analyte can be recovered outside the body.

[0030] As used herein, the terms "permeant", "drug", "permeant composition", or "pharmacologically active agent" or any other similar term are used interchangeably to refer to any chemical or biological material or compound suitable for transdermal administration by methods previously known in the art and / or methods taught herein that can induce a desired biological or pharmacological effect, including but not limited to (1) having a prophylactic effect on an organism and preventing undesired biological effects such as infections, (2) alleviating a condition caused by a disease, such as alleviating pain or inflammation, and / or (3) alleviating, reducing, or completely eliminating a disease from an organism. The effect may be local, such as providing a local anesthetic effect, or systemic. Such substances include a wide class of compounds that are normally delivered in the body, including those through body surfaces and membranes including the skin. Generally, by way of example and not limitation, such substances can include any physiologically active agent such as a drug, chemical, or biological material that induces a desired biological or pharmacological effect. For this purpose, in one aspect, the permeant can be a low molecular weight agent. In another aspect, the permeant can be a high molecular weight agent.

[0031] In various embodiments, systems, devices, and methods for use and / or adapted for use with the compositions and methods described herein are described in U.S. Patent Nos. 6,022,316; 6,142,939; 6,173,202; 6,183,434; 6,508,785; 6,527,716; 6,692,456; 6,730,028; 7,141,034; 7,392,080; 7,758,561; 8,016,811; 8,116,860; and / or 9,498,609, all of which are hereby incorporated by reference in their entirety for the purpose of particularly describing such systems and methods. In various embodiments, systems and devices commercially available from PASSPORT® may be used or adapted for use in the delivery of the compositions described herein.

[0032] As used herein, an "effective" amount of a pharmacologically active agent means an amount of the compound sufficient to provide the desired local or systemic effects and performance with a reasonable benefit / risk ratio attendant to any medical treatment. An "effective" amount of a penetration or chemical enhancer as used herein means an amount selected to provide the desired increase in biological membrane permeability, desired depth of penetration, rate of administration, and amount of drug delivered.

[0033] As used herein, "animal" or "organism" refers to humans to whom the present disclosure may apply, and other organisms including plants.

[0034] As used herein, "analyte" means any chemical or biological material or compound suitable for passing through a biological membrane that an individual may wish to know the concentration or activity of in their body, by the techniques taught in this disclosure or by techniques previously known in the art. Glucose is a specific example of an analyte, as it is a sugar suitable for passing through the skin, and an individual, such as one suffering from diabetes, may wish to know their blood glucose level. Other examples of analytes include, but are not limited to, compounds such as sodium, potassium, bilirubin, urea, ammonia, calcium, lead, iron, lithium, salicylate, and the like.

[0035] As used herein, "transdermal flux rate" is the rate of passage of any analyte through the skin of an individual, human or animal, or the rate of passage of any permeant, drug, pharmacologically active agent, dye or pigment within and through the skin of an organism.

[0036] As used herein, "non-invasive" means not requiring the entry of a needle, catheter, or other invasive medical device into a part of the body.

[0037] As used herein, "minimally invasive" refers to the use of mechanical, hydraulic, or electrical means to penetrate the stratum corneum and create small pores or micropores without causing substantial damage to the underlying tissue.

[0038] As used herein, "pharmaceutically acceptable carrier" refers to a carrier into which a pharmaceutically acceptable substance such as a drug can be provided for delivery. Pharmaceutically acceptable carriers are described in the art, for example, in "Remington: The Science and Practice of Pharmacy", Mack Publishing Company, Pennsylvania, 1995, the disclosure of which is incorporated herein by reference. Carriers can include, for example, water and other aqueous solutions, sugars, polysaccharides, buffers, excipients, and biodegradable polymers such as polyesters, polyanhydrides, polyamino acids, liposomes, and mixtures thereof.

[0039] As used herein, "reservoir" refers to a designated area or chamber within a device that is designed to contain an osmotic agent for delivery to an organism through an artificial opening in a biological membrane or to receive a biological fluid sample extracted from an organism through an artificial opening in a biological membrane. The reservoir can also contain excipient compounds that enhance the effect of separately contained bioactive osmotic agents. Additionally, the reservoir can contain or be treated with reactive enzymes or reagents designed to enable the measurement or detection of selected analytes in the extracted biological fluid. The reservoir can include an open volume space, a gel, a flat planar space coated or treated with a compound selected for subsequent release or reaction, or a matrix or permeable solid structure such as a pellet, tablet, powder, dry solid, or porous polymer.

[0040] As used herein, "p3-Alcβ" is produced as a metabolite of the membrane protein alcadein β (Alcβ). Alcβ is metabolized via a pathway similar to that of Aβ to produce p3-Alcβ. Unlike Aβ that accumulates in the brain, p3-Alcβ is more significantly decreased in Alzheimer's disease patients. The actions of p3-Alcβ are performed by its partial peptides, such as p3-Alcβ1-40, p3-Alcβ1-37, p3-Alcβ9-19, p3-Alcβ1-19, p3-Alcβ11-19, or derivatives thereof.

[0041] Patch In some embodiments, the systems, devices, and methods of the present disclosure can be used to transdermally deliver peptides across the skin. In some embodiments, the patch can include an upper layer that includes an adhesive, an intermediate layer that includes a matrix, and a lower layer. In some embodiments, the lower layer includes a release liner. In some embodiments, the intermediate layer further includes a PK modifier. In some embodiments, the intermediate layer further includes an enhancer. In some embodiments, the patch includes a tissue interface layer. In some embodiments, the patch further includes a backing. In some embodiments, the release liner is configured to be removed before application to the skin of the subject. In some embodiments, the patch is configured as a film. In some embodiments, the patch is configured as a pellet.

[0042] Examples of suitable tissue interface layers are described in U.S. Patent No. 7,392,080, which is hereby incorporated by reference in its entirety for the purpose of particularly describing transdermal drug delivery patch systems.

[0043] In some embodiments, the upper layer includes a backing. In some embodiments, the backing is a film, foam, woven fabric, or non-woven material. In some embodiments, the film includes polyethylene (PE), polyethylene terephthalate (PET), polyurethane (PU), polyvinyl chloride (PVC), polychlorotrifluoroethylene (PCTFE), cyclic olefin copolymer (COC), or cyclic olefin polymer (COP). In some embodiments, the backing includes an adhesive. In some embodiments, the adhesive is an acrylic rubber, silicone rubber, or a synthetic rubber such as polyisobutylene (PIB) and styrene-isoprene-styrene block copolymer (SIS). In some embodiments, the color of the backing is transparent, translucent, tan, white, or beige. In some embodiments, the backing is formed by thermoforming to create cavities. In some embodiments, the backing is covered with adhesive tape.

[0044] In some embodiments, the lower layer includes a release liner. In some embodiments, the release liner is a film. In some embodiments, the film is polyethylene terephthalate, polyethylene, paper, or aluminum foil. In some embodiments, the film includes a silicone or fluorosilicone coating layer. In some embodiments, the release liner is heat-sealed to a backing film.

[0045] In some embodiments, the PK regulator is a drug delivery regulator. In some embodiments, the PK regulator is a pH control agent. In some embodiments, the PK regulator is an organic acid, a salt form of an organic acid, or a combination thereof. In some embodiments, the organic acid is selected from ascorbic acid, citric acid, succinic acid, tartaric acid, maleic acid, lactic acid, benzoic acid, sorbic acid, amino acids, or a combination thereof. In some embodiments, the PK regulator is an inorganic acid. In some embodiments, the PK regulator is a salt form of an inorganic acid. In some embodiments, the inorganic acid is hydrochloric acid, phosphoric acid, boric acid, acetic acid, or a combination thereof. In some embodiments, the inorganic acid evaporates during the manufacturing process. In some embodiments, the organic base is selected from sodium citrate, tris, monosodium phosphate, disodium phosphate, trisodium phosphate, monopotassium phosphate, dipotassium phosphate, tripotassium phosphate, basic amino acids, or a combination thereof.

[0046] In some embodiments, the enhancer is a saccharide. In some embodiments, the saccharide comprises a sugar or is a sugar. In some embodiments, the enhancer is a non-reducing saccharide. In some embodiments, the enhancer is a reducing saccharide. In some embodiments, the saccharide is selected from mannitol, maltose, trehalose, xylitol, xylose, dextrose, lactose, sorbitol, sucrose, fructose, maltitol, erythritol, lactitol, isomalt, and cyclodextrin, or a combination thereof. In some embodiments, the enhancer is sucrose. In some embodiments, the enhancer is lactose. In some embodiments, the enhancer is maltose. In some embodiments, the enhancer is a combination of sucrose and lactose.

[0047] In some embodiments, the weight ratio of the enhancer to the non-aggregated peptide is greater than 0.005, greater than 0.02, greater than 0.05, greater than 0.1, greater than 0.2, or in a range including and / or extending to the aforementioned values. In some embodiments, the weight ratio of the sugar to the non-aggregated peptide is greater than 0.02. In some embodiments, the weight ratio of the sugar to the non-aggregated peptide is from about 0.02 to about 10. In some embodiments, the weight ratio of the sugar to the non-aggregated peptide is from about 0.02 to about 0.4.

[0048] In some embodiments, the intermediate layer of the patch comprises a reservoir. In some embodiments, the reservoir comprises from about 1.0 wt% to about 99.5 wt% of the non-aggregated peptide. In some embodiments, the non-aggregated peptide comprises approximately 50 wt%, 55 wt%, 70 wt%, 80 wt%, 90 wt%, 95 wt%, 98 wt%, etc. of the intermediate layer, including approximately 50 wt% to approximately 98 wt% of the intermediate layer, including any range of weight percentages derivable from these values.

[0049] In some embodiments, the intermediate layer comprises from about 0.5 wt% to about 99 wt% enhancer. In some embodiments, the enhancer comprises from about 2 wt% to about 50 wt% of the intermediate layer, including amounts such as 2 wt%, 5 wt%, 10 wt%, 15 wt%, 30 wt%, 40 wt%, 50 wt% of the intermediate layer, including any range of weight percentages derivable from these values.

[0050] In some embodiments, the intermediate layer comprises from about 5 wt% to about 60 wt% PK regulator. In some embodiments, the PK regulator comprises from about 10 wt% to about 40 wt% of the intermediate layer, including additional amounts such as 7.5 wt%, 15 wt%, 25 wt%, 40 wt% of the intermediate layer, including any range of weight percentages derivable from these values.

[0051] In some embodiments, the intermediate layer comprises at least 0.5 mg / cm 2 , at least 1 mg / cm 2 , at least 2 mg / cm 2 , at least 4 mg / cm 2 , at least 8 mg / cm 2 , or an area amount of PK regulator in a range including and / or extending to the foregoing values, based on the surface area of the intermediate layer facing the lower layer. In some embodiments, the intermediate layer comprises at least 2 mg / cm 2 of disodium citrate. In some embodiments, the intermediate layer comprises at least 4 mg / cm 2 of disodium citrate.

[0052] In some aspects, the intermediate layer comprises a matrix support. In some embodiments, the matrix support comprises at least one fiber. In some embodiments, the fiber is a nonwoven material. In some embodiments, the matrix support is a nonwoven fabric. In some embodiments, the nonwoven fabric is polyethylene terephthalate. In some embodiments, the matrix support is a laminated material of a film. In some embodiments, the film is polyethylene terephthalate. In some embodiments, the matrix support is a laminated material of fibers. In some embodiments, the matrix is a laminated material of a film and fibers.

[0053] In some embodiments, the thickness of the matrix support is less than 300 μm, less than 250 μm, less than 200 μm, less than 150 μm, less than 100 μm, less than 50 μm, or in a range including and / or extending to the aforementioned values.

[0054] In some embodiments, the areal weight of the fiber is less than 100 g / m 2 2, less than 90 g / m 2 2, less than 80 g / m 2 2, less than 70 g / m 2 2, less than 60 g / m 2 2, less than 50 g / m 2 2, less than 40 g / m 2 2, less than 30 g / m 2 2, less than 20 g / m 2 2, less than 10 g / m 2 2, or in a range including and / or extending to the aforementioned values.

[0055] In some embodiments, the matrix support has a water holding capacity (WHC) of about 0.1 mg / cm 2 2 to about 10 mg / cm 2 2 based on the surface area of the matrix support facing the lower layer. The water holding capacity of the matrix support means the amount of water that the matrix support can hold per 1 cm 2 2 of the transdermal surface. Specifically, a 1 cm 2 2 matrix is prepared and immersed in a solution (phosphate buffered saline containing 0.1% surfactant (Tween® 80)) for a sufficiently long time. Then, the matrix support is slowly withdrawn from the solution for about 5 seconds, and the weight of the sample before immersion measured in advance is subtracted from the weight of the sample holding the liquid, whereby the water holding capacity of the matrix per unit area (1 cm 2 2) of the transdermal surface can be determined. In some embodiments, the water holding capacity of the matrix support is about 10 mg / cm 2 2 or less. In some embodiments, the water holding capacity of the matrix support is about 1 mg / cm 2 2 to about 8 mg / cm 2It is. In some embodiments, the water retention capacity of the matrix is about 2 mg / cm 2 ~ about 5 mg / cm 2 .

[0056] The water retention capacity of the matrix support can be controlled by adjusting the thickness and weight of the matrix support. The matrix support preferably has a thickness of 100 μm or less. In some embodiments, the matrix support has a thickness in the range of about 10 μm to about 100 μm. In some embodiments, the thickness of the matrix support is about 20 μm to about 90 μm. In some embodiments, the thickness of the matrix support is about 30 μm to about 80 μm. In some embodiments, the thickness of the matrix support is about 40 μm to about 60 μm.

[0057] In some embodiments, the basis weight of the matrix support is about 10 g / m 2 ~ about 100 g / m 2 . In some embodiments, the basis weight of the matrix support is about 15 g / m 2 ~ about 80 g / m 2 . In some embodiments, the basis weight of the matrix support is about 20 g / m 2 ~ about 60 g / m 2 . In some embodiments, the basis weight of the matrix support is about 25 g / m 2 ~ about 40 g / m 2 .

[0058] In some embodiments, the basis weight of the matrix support is about 0.1 mg / cm 2 ~ about 30 mg / cm 2 . In some embodiments, the basis weight of the matrix support is about 0.5 mg / cm 2 ~ about 30 mg / cm 2 . In some embodiments, the basis weight of the matrix support is about 0.5 mg / cm 2 ~ about 20 mg / cm 2 . In some embodiments, the basis weight of the matrix support is about 0.5 mg / cm 2 ~ about 10 mg / cm 2 .

[0059] In some embodiments, the size of the matrix is about 0.125 cm 2 to about 4 cm 2 In some embodiments, the size of the matrix is about 0.25 cm 2 to about 3 cm 2 In some embodiments, the size of the matrix is about 0.5 cm 2 to about 2 cm 2 In some embodiments, the size of the matrix is about 0.5 cm 2 In some embodiments, the size of the matrix is about 1 cm 2 In some embodiments, the size of the matrix is about 2 cm 2 In some embodiments, the size of the matrix is about 2 cm

[0060] In some embodiments, the total amount of non-aggregated peptide and enhancer per unit area (surface area) of the matrix is 0.01 mg / cm 2 to 200 mg / cm 2 In some embodiments, the total amount of non-aggregated peptide and enhancer per unit area of the matrix is 0.1 mg / cm 2 to 100 mg / cm 2 In some embodiments, the total amount of non-aggregated peptide and enhancer per unit area of the matrix is 5 mg / cm 2 to 75 mg / cm 2 In some embodiments, the total amount of non-aggregated peptide and enhancer per unit area of the matrix is 10 mg / cm 2 to 50 mg / cm 2 In some embodiments, the total amount of non-aggregated peptide and enhancer per unit area of the matrix is 10 mg / cm

[0061] In some embodiments, the pH of the matrix component is from about 3 to about 9. In some embodiments, the pH of the matrix is from about 4 to about 8. In some embodiments, the pH of the matrix is about 4. In some embodiments, the pH of the matrix is about 5. In some embodiments, the pH of the matrix is about 6. In some embodiments, the pH of the matrix is about 7.

[0062] In some embodiments, the matrix contains non-aggregated peptides at about 0.01 mg / cm 2 to about 200 mg / cm 2 based on the surface area of the matrix facing the lower layer. In some embodiments, the matrix contains non-aggregated peptides at about 0.1 mg / cm 2 to about 100 mg / cm 2 based on the surface area of the matrix facing the lower layer. In some embodiments, the matrix contains non-aggregated peptides at about 1 mg / cm 2 to about 50 mg / cm 2 based on the surface area of the matrix facing the lower layer. In some embodiments, the matrix contains non-aggregated peptides at about 5 mg / cm 2 to about 30 mg / cm 2 based on the surface area of the matrix facing the lower layer. In some embodiments, the matrix contains non-aggregated peptides at about 5 mg / cm 2 to about 25 mg / cm 2 based on the surface area of the matrix facing the lower layer. In some embodiments, the matrix contains non-aggregated peptides at about 0.5 mg / cm 2 based on the surface area of the matrix facing the lower layer. In some embodiments, the matrix contains non-aggregated peptides at about 1 mg / cm 2 based on the surface area of the matrix facing the lower layer. In some embodiments, the matrix contains non-aggregated peptides at about 2 mg / cm 2 based on the surface area of the matrix facing the lower layer. In some embodiments, the matrix contains non-aggregated peptides at about 3 mg / cm 2 based on the surface area of the matrix facing the lower layer. In some embodiments, the matrix contains non-aggregated peptides at about 5 mg / cm 2 based on the surface area of the matrix facing the lower layer. In some embodiments, the matrix contains non-aggregated peptides at about 10 mg / cm 2 based on the surface area of the matrix facing the lower layer. In some embodiments, the matrix contains non-aggregated peptides at about 20 mg / cm 2 based on the surface area of the matrix facing the lower layer. In some embodiments, the matrix contains non-aggregated peptides at about 25 mg / cm 2 based on the surface area of the matrix facing the lower layer. In some embodiments, the matrix contains non-aggregated peptides at about 50 mg / cm 2It contains a non-aggregating peptide. In some embodiments, the non-aggregating peptide is p3-Alcβ. In some embodiments, p3-Alcβ is selected from at least one of p3-Alcβ1-40, p3-Alcβ1-37, p3-Alcβ9-19, p3-Alcβ1-19, p3-Alcβ11-19, or a derivative thereof. In some embodiments, p3-AlcB is p3-Alcβ9-19 or a derivative thereof.

[0063] In some aspects, the matrix is about 0.05 mg / cm 2 ~ about 100 mg / cm 2 of enhancer. In some embodiments, the matrix is about 0.1 mg / cm 2 ~ about 50 mg / cm 2 of enhancer. In some embodiments, the matrix is about 0.2 mg / cm 2 ~ about 5.0 mg / cm 2 of enhancer. In some embodiments, the matrix is about 0.5 mg / cm 2 ~ about 10 mg / cm 2 of enhancer. In some embodiments, the matrix is about 0.5 mg / cm 2 ~ about 4.0 mg / cm 2 of enhancer. In some embodiments, the matrix is about 0.5 mg / cm 2 ~ about 2.0 mg / cm 2 of enhancer. In some embodiments, the matrix is about 0.5 mg / cm 2 of enhancer. In some embodiments, the matrix is about 1 mg / cm 2 of enhancer. In some embodiments, the matrix is about 2 mg / cm 2 of enhancer. In some embodiments, the enhancer is selected from at least sucrose, lactose, and maltose.

[0064] In some embodiments, the patch further comprises an antimicrobial agent. In some embodiments, the antimicrobial agent is selected from benzoic acid, methylparaben, propylparaben, benzalkonium chloride, chlorhexidine, cresol, salicylic acid, sorbic acid, sodium benzoate, benzethonium chloride, and combinations thereof.

[0065] In some embodiments, the patch is configured as a dry patch formulation. In some embodiments, the dry patch comprises a backing, a matrix containing a non-aggregated peptide in a dry state, and a release liner. In some embodiments, the matrix further comprises a PK regulator in a dry state. In some embodiments, the matrix further comprises an enhancer in a dry state. In some embodiments, the matrix further comprises an antimicrobial agent in a dry state. In some embodiments, the dry patch is a heat-dried film produced by a dispensing or casting process. In some embodiments, the dry patch is a tablet or pellet produced by a compression process. In some embodiments, the dry patch formulation comprises p3-Alcβ9-19, sucrose, lactose, disodium citrate sesquihydrate, methylparaben, propylparaben, sodium benzoate, and benzalkonium chloride. In some embodiments, the dry state contains less than 10% water in the formulation. In some embodiments, the dry state contains less than 5% water in the formulation. In some embodiments, the dry state contains less than 2% water in the formulation. In some embodiments, the dry state contains less than 1% water in the formulation.

[0066] In some embodiments, the patch is configured as a lyophilized patch formulation. In some embodiments, the lyophilized patch comprises a backing, a matrix comprising p3-Alcβ9-19 in a lyophilized state, and a release liner. In some embodiments, the lyophilized patch consists of a cavity having a spacer. In some embodiments, the lyophilized patch consists of a blister package. In some embodiments, the matrix further comprises a drug delivery regulator in a lyophilized state. In some embodiments, the matrix comprises an enhancer in a lyophilized state. In some embodiments, the matrix further comprises an antimicrobial agent in a lyophilized state. In some embodiments, the lyophilized patch is lyophilized by methods known to those skilled in the art. In some embodiments, the lyophilized patch is a tablet or pellet produced by a compression process. In some embodiments, the lyophilized patch formulation comprises p3-Alcβ9-19, sucrose, lactose, disodium citrate sesquihydrate, methylparaben, propylparaben, sodium benzoate, and benzalkonium chloride. In some embodiments, the lyophilized patch contains less than 10% water in the formulation. In some embodiments, the lyophilized patch contains less than 5% water in the formulation. In some embodiments, the lyophilized patch contains less than 2% water in the formulation. In some embodiments, the lyophilized patch contains less than 1% water in the formulation. In some embodiments, the lyophilized patch contains less than 0.5% water in the formulation. In some embodiments, the lyophilized patch contains less than 0.1% water in the formulation.

[0067] In some embodiments, the patch is configured as a tablet patch. In some embodiments, the tablet patch is a thin solid tablet. The thin solid tablet can be in various wafer or plate-like shapes such as oval, circular, triangular, rectangular, square, pentagonal, hexagonal, irregular, etc. In various embodiments, the thin solid tablet is substantially flat. In one embodiment, the substantially flat thin solid tablet is slightly bent or curved to an extent that facilitates handling as compared to, for example, a flat thin solid tablet that is more difficult to pick up from a flat surface. In various embodiments, the thin solid tablets described herein are 30 mg / cm 2Greater than 40 mg / cm 2 Greater than 50 mg / cm 2 Greater than 60 mg / cm 2 Greater than 70 mg / cm 2 Greater than 80 mg / cm 2 Greater than 90 mg / cm 2 Greater than or 100 mg / cm 2 Greater than; 400 mg / cm 2 Less than, 350 mg / cm 2 Less than, 300 mg / cm 2 Less than, 250 mg / cm 2 Less than, or 200 mg / cm 2 Less than; or have areal density in any range defined by any two of the foregoing values and having an end point. For example, in various embodiments, the thin solid tablets have areal density greater than 30 mg / cm 2 Greater than 400 mg / cm 2 Less than; 40 mg / cm 2 Greater than 400 mg / cm 2 Less than; or 30 mg / cm 2 Greater than 400 mg / cm 2 Having an areal density less than.

[0068] In various embodiments, the thin solid tablets described herein have a thickness of about 0.01 mm or more, about 0.02 mm or more, about 0.03 mm or more, about 0.04 mm or more, about 0.05 mm or more, about 0.1 mm or more, about 0.2 mm or more, about 0.5 mm or more, or about 1 mm or more; about 10 mm or less, about 5 mm or less, about 2 mm or less, or about 1 mm or less; or have a thickness in any range defined by any two of the foregoing values and having an end point. For example, in various embodiments, the thin solid tablets have a thickness in the range of about 0.01 mm to about 10 mm or in the range of about 0.1 mm to about 5 mm.

[0069] In various embodiments, the thin solid tablets have a surface in a pattern similar to the front or back of a coin. In various embodiments, the surface of the thin solid tablets is about 0.01 cm 2 or more, about 0.05 cm 2 or more, about 0.1 cm 2 or more, about 0.25 cm2 Above, about 0.5 cm 2 Above, about 0.75 cm 2 Above, or about 1 cm 2 Above; or about 50 cm 2 Below, about 25 cm 2 Below, about 15 cm 2 Below, about 10 cm 2 Below, about 5 cm 2 Below, or about 2 cm 2 Below, or has an area of any range having endpoints defined by any two of the aforementioned values. For example, in various embodiments, the surface of the thin solid tablet is about 0.01 cm 2 ~ about 25 cm 2 、about 0.1 cm 2 ~ about 10 cm 2 、or about 0.15 cm 2 ~ about 5 cm 2 and has an area within this range.

[0070] In some embodiments, the patch is configured as a reservoir patch. In some embodiments, the reservoir patch includes a backing, a spacer for creating a cavity, a matrix containing p3 - Alcβ9 - 19 within the cavity, and a release liner. The matrix can be of various shapes such as elliptical, circular, triangular, rectangular, square, pentagonal, hexagonal, irregular, etc. In certain embodiments, the depth of the cavity is about 0.5 mm to 10 mm. In some embodiments, the depth of the cavity is about 0.5 mm to 5 mm. In some embodiments, the depth of the cavity is 1 mm to 3 mm.

[0071] In some embodiments, the patch is configured as a lyophilized dry patch. In some embodiments, the lyophilized dry patch includes a backing, an intermediate layer containing a matrix of lyophilized p3 - Alcβ9 - 19, and a release liner. The matrix can be of various shapes such as elliptical, circular, triangular, rectangular, square, pentagonal, hexagonal, irregular, etc. In certain embodiments, the matrix is about 0.5 mm to 10 mm. In some embodiments, the matrix is about 0.5 mm to 5 mm. In some embodiments, the matrix is 1 mm to 3 mm.

[0072] In some embodiments, the patch is configured as a tablet patch. In some embodiments, the tablet patch includes a backing, an intermediate layer including a tablet containing p3-Alcβ9-19, and a release liner. The matrix can be of various shapes such as oval, circular, triangular, rectangular, square, pentagonal, hexagonal, irregular, etc. In certain embodiments, the matrix is about 0.5 mm to 10 mm. In some embodiments, the tablet is about 0.5 mm to 5 mm. In some embodiments, the tablet is 1 mm to 3 mm.

[0073] In some embodiments, the patch is configured as a solid dispersion dry patch. In some embodiments, the solid dispersion dry patch includes a backing, an intermediate layer including a matrix containing p3-Alcβ9-19 in a dry state, and a release liner. The matrix can be of various shapes such as oval, circular, triangular, rectangular, square, pentagonal, hexagonal, irregular, etc. In certain embodiments, the matrix is about 0.5 mm to 10 mm. In some embodiments, the tablet is about 0.5 mm to 5 mm. In some embodiments, the matrix is 1 mm to 3 mm.

[0074] Method / Use Aspects disclosed herein relate to administering to a subject in need of an effective amount of a non-aggregated peptide disclosed elsewhere herein. Some embodiments relate to treating Alzheimer's disease through administration of a patch disclosed herein. In some embodiments, a patch of the present disclosure can transdermally deliver a non-aggregated peptide to a subject through one or more micropores formed by a transdermal delivery system described herein. In some embodiments, the method can include perforating the outermost stratum corneum and the epidermis such that the non-aggregated peptide in the patch passes through the epidermis, diffuses into the dermal capillaries, and enters the systemic circulation, and then applying the patch thereon. The non-aggregated peptide absorbed from the skin can avoid first-pass metabolism in the liver and can be effectively delivered to the brain. Thus, embodiments of the patch of the present disclosure can provide a drug administration means alternative to injection that can be suitably used for immediate release administration or sustained release of a non-aggregated peptide with high bioavailability compared to oral, nasal, or passive transdermal administration.

[0075] In some embodiments, a patch of the present disclosure can also be used in a method of delivering a non-aggregated peptide through a target biological membrane, the method including steps for forming one or more micropores in the biological membrane and steps for placing the patch in physical contact with the one or more micropores such that at least a portion of the drug becomes soluble in the biological moisture received from the target through the one or more micropores.

[0076] In some embodiments, a method of treating a subject includes steps for identifying a subject having a brain disease or brain condition, steps for opening a plurality of micropores in the subject's skin, and steps for applying a patch over the micropores of the subject's skin for a period effective to effect transdermal delivery of a non-aggregated peptide. In some embodiments, the patch includes an upper layer including an adhesive, an intermediate layer including a non-aggregated peptide, and a lower layer. In some embodiments, the lower layer includes a release liner. In some embodiments, the period is selected to deliver a therapeutically effective amount of the non-aggregated peptide through the plurality of micropores.

[0077] In some embodiments, opening a plurality of micropores in the skin of a subject comprises applying a transdermal microporation device to the skin of the subject. In some embodiments, the transdermal microporation device comprises a conductive member including an array of conductive filaments. In some embodiments, the transdermal microporation device comprises a conductive member including an array of conductive filaments. In some embodiments, transdermal microporation opens the micropores by thermal tissue ablation. In some embodiments, transdermal microporation generates micropores through the stratum corneum to the epidermis.

[0078] The patch of the present disclosure can be applied to a subject having a disease or condition for which an immediate or sustained effect is desired. In some embodiments, the patch and transdermal delivery of the non-aggregated peptide can have a PK profile comparable to subcutaneous injection. In some embodiments, the patch and transdermal delivery of the non-aggregated peptide can have a PK profile superior to intravenous and subcutaneous injection. In some embodiments, the patch and transdermal delivery of the non-aggregated peptide can have a bioavailability of more than 100%, more than 200%, more than 300%, more than 400%, more than 500%, more than 750%, more than 1000%, more than 1500%, more than 2000%, more than 2500%, more than 3000%, more than 3500%, more than 4000%, more than 4500%, more than 5000%, or including and / or ranging between the aforementioned values as compared to intravenous (IV) administration. In some embodiments, the patch and transdermal delivery of the non-aggregated peptide can have a PK profile including a bioavailability of about 350% to about 4800% as compared to IV.

[0079] In some embodiments, a patch and transdermal delivery of a non-aggregating peptide can provide enhanced delivery of the non-aggregating peptide across the skin of a subject. In some embodiments, a patch and transdermal delivery of a non-aggregating peptide can provide longer sustained delivery of the non-aggregating peptide to a subject as compared to intravenous or subcutaneous administration. In some embodiments, the area under the curve (AUC) of a patch and transdermal delivery of a non-aggregating peptide in plasma for the non-aggregating peptide is about 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0 times higher than that of an intravenous injection, or is in a range including and / or spanning the aforementioned values. In some embodiments, the area under the curve (AUC) of a patch and transdermal delivery of a non-aggregating peptide in plasma for the non-aggregating peptide is 5.4 times higher than that of an intravenous injection. The area under the curve (AUC) of a patch and transdermal delivery of a non-aggregating peptide in plasma for the non-aggregating peptide is 8.9 times higher than that of an intravenous injection.

[0080] In some embodiments, a patch and transdermal delivery of an effective amount of a non-aggregating peptide into the blood of a subject is achieved at at least about 0.5 hours after administration, about 1 hour after administration, about 2 hours after administration, about 3 hours after administration, about 4 hours after administration, about 5 hours after administration, about 6 hours after administration, about 7 hours after administration, about 8 hours after administration, or in a range including and / or spanning the aforementioned values.

[0081] In some embodiments, a patch and transdermal delivery of an effective amount of a non-aggregating peptide into the blood of a subject is maintained for more than 3 hours after administration, more than 4 hours after administration, more than 5 hours after administration, more than 6 hours after administration, more than 7 hours after administration, more than 8 hours after administration, more than 9 hours after administration, more than 10 hours after administration, or in a range including and / or spanning the aforementioned values.

[0082] In some embodiments, a patch and transdermal delivery of a non-aggregating peptide into cerebrospinal fluid achieves a Tmax at about 1 hour after administration, about 2 hours after administration, about 3 hours after administration, about 4 hours after administration, about 5 hours after administration, about 5 hours after administration, or in a range including and / or spanning the aforementioned values.

[0083] In some embodiments, the movement of the non-aggregated peptide from blood to cerebrospinal fluid (expressed as a percentage, the amount of non-aggregated peptide in cerebrospinal fluid divided by the amount in plasma) is about 1.0%, about 1.5%, about 2.0%, about 3.0%, about 3.5%, about 4.0%, about 4.5%, about 5.0%, about 5.5%, about 6.0%, or a range that includes and / or extends between the foregoing values. In some embodiments, the movement of the non-aggregated peptide from blood to cerebrospinal fluid is about 3.5%.

[0084] In some embodiments, methods for activating mitochondria in the brain are provided. In some embodiments, the method includes the step of opening at least one microchannel in the skin of a subject and the step of applying a patch disclosed herein to the skin of the subject. In some embodiments, activation of BCPP-EF accumulation in mitochondria is achieved at 0.5 hour after administration, 1.0 hour after administration, 1.5 hours after administration, 2.0 hours after administration, 2.5 hours after administration, 3.0 hours after administration, 3.5 hours after administration, 4.5 hours after administration, 5.0 hours after administration, or a range that includes and / or extends between the foregoing values. In some embodiments, activation of mitochondria in the brain is maintained for more than 3 hours after administration, more than 4 hours after administration, more than 5 hours after administration, more than 6 hours after administration, more than 7 hours after administration, more than 8 hours after administration, more than 9 hours after administration, more than 10 hours after administration, or a range that includes and / or extends between the foregoing values.

[0085] In some embodiments, the subject receives sufficient non-aggregated peptide from multiple administrations before a high level of non-aggregated peptide is achieved. One can readily and immediately envision a regimen where the subject is administered a first patch and the subject receives one or more subsequent patches. Such a regimen can continue such that the subject receives a third patch after receiving a second patch. In some embodiments, the subject can receive 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more patches during the treatment. One or more additional patches described herein can be administered before the first patch administration or before one or more subsequent patch administrations. In some embodiments, the subject is administered over the remainder of their lifetime and / or over a number of years (e.g., 1 year or more, 5 years or more, 10 years or more, 15 years or more, 20 years or more, 30 years or more, or ranges including and / or extending between the foregoing values).

[0086] In some cases, a period of time elapses while one or more patches are being administered to the subject. In some embodiments, the period of time while one or more patches are being administered is equal to or at least approximately 1 day twice a day, 1 day, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, or ranges including and / or extending between the foregoing values. In some embodiments, one or more additional therapeutic agents are administered to the subject during the period while the composition is being administered to the subject.

[0087] Subjects in need of receiving the patches disclosed herein to improve the health of the subject do not always need to be identified prior to receiving the first treatment with the patches described herein. For example, a subject may be determined in advance to develop Alzheimer's disease in the future before showing any current signs or symptoms of Alzheimer's disease (cognitively normal in the preclinical stage) or mild cognitive impairment of Alzheimer's disease (mild MCI stage). Alternatively, a subject may be prophylactically treated when there is or is not a risk of Alzheimer's disease (e.g., when the patient reaches an age of 50 or older, 60 or older, 70 or older, etc.). Thus, in some embodiments, the patch is administered to the subject after the subject has received an early diagnosis. In some embodiments, not all subjects are candidates for such administration, and it may be desirable to identify subjects for treatment. It is understood that the selection of patients depends on several factors within the skill of a physician who is a person of ordinary skill in the art. Accordingly, some embodiments disclosed herein identify subjects as those who would benefit from administering an effective amount of at least one non-aggregated peptide or a composition comprising the same, and further comprise the step of prolonging lifespan, extending survival time, extending life expectancy, or improving by immunization. The subject may be identified based on the subject's age, current medical condition, current medical treatment, in accordance with the prescribed medical treatment, or in some embodiments, physiological factors specific to a subject diagnosed with Alzheimer's disease. In some embodiments, treatment of Alzheimer's disease includes preventing, reducing, and / or delaying the accumulation of tangles of beta-amyloid, amyloid plaques, and / or tau protein. Beta-amyloid is the remaining fragment of a larger protein. When these fragments cluster together, they appear to have a toxic effect on neurons and disrupt cell-to-cell communication. These clusters form larger deposits called amyloid plaques that also contain other cellular debris. Tau protein serves as part of the internal support and transport system of neurons to carry nutrients and other essential substances. In Alzheimer's disease, the tau protein changes shape and organizes into structures called neurofibrillary tangles. Neurofibrillary tangles disrupt the transport system and are toxic to cells.

[0088] In some embodiments, provided is a method of treating, preventing, or ameliorating a brain disease by administering a patch as described herein. In some embodiments, the brain disease is dementia. In some embodiments, the dementia is caused by Alzheimer's disease and Parkinson's disease. Dementia is a broad category of brain diseases that cause a long-term and often progressive decline in the ability to think and remember that is severe enough to affect daily functioning. Symptoms associated with dementia include emotional problems, language disorders, and reduced motivation. In some embodiments, the brain disease is a neurodegenerative disease such as amyotrophic lateral sclerosis (ALS), Huntington's disease, ataxia, spinal muscular atrophy, Lewy body disease, and the like. In some embodiments, the method includes testing a subject for dementia risk factors and administering a composition as described herein. In some embodiments, testing a subject for dementia risk factors includes performing a brain scan and amyloid PET, performing a brain biopsy, performing a cognitive test, and testing the subject's blood. The subject can be identified based on the subject's age, current medical condition, current medical treatments, physiological factors specific to the subject, or in some embodiments, a subject diagnosed with dementia, in accordance with the prescribed medical treatments.

[0089] Some aspects provided herein provide methods for producing supra-physiologically high levels of non-aggregated peptides in a subject. In some embodiments, the method includes providing to the subject a non-aggregated peptide, composition, or patch disclosed elsewhere herein. In some embodiments, the method includes administering an effective amount of a non-aggregated peptide disclosed herein. In some embodiments, as disclosed herein, the composition includes a non-aggregated peptide, an enhancer, a PK regulator, and an antimicrobial agent. In some embodiments, the enhancer is a reducing sugar. In some embodiments, the PK regulator is an organic acid. In some embodiments, the antimicrobial agent is parabens. In some embodiments, the antimicrobial agent is sodium benzoate. In some embodiments, the patch increases the bioavailability of the non-aggregated peptide by at least 200%. In some embodiments, the method provides a transfer rate of the non-aggregated peptide from blood to CSF of about 3.5%.

[0090] Some aspects are methods for treating or improving a disease, disorder, or condition associated with Alzheimer's disease in a subject, the method comprising administering to the subject a therapeutically effective amount of a non-aggregated peptide, composition, or patch disclosed elsewhere herein. In some embodiments, the patch includes a non-aggregated peptide, an enhancer, a PK regulator, and an antimicrobial agent. In some embodiments, the non-aggregated peptide or composition improves cerebral blood flow in the subject. In some embodiments, the non-aggregated peptide or composition prevents or inhibits hyperphosphorylation of tau protein. In some embodiments, hyperphosphorylation of tau protein is inhibited by 50%, 70%, 80%, 90%, 99%, or equal to or at least approximately those values in a range including and / or extending between the foregoing values. In some embodiments, the composition reduces beta-amyloid in the subject's brain. In some embodiments, beta-amyloid in the subject's brain is reduced by 50%, 70%, 80%, 90%, 99%, or equal to or at least approximately those values in a range including and / or extending between the foregoing values. In some embodiments, the aggregated peptide or composition has neuroprotective and neuroactivating properties.

[0091] In some embodiments, the treatment method described herein down-regulates the β-secretase activity of a subject. In some embodiments, the β-secretase activity decreases by only or at least approximately those values equal to 50%, 70%, 80%, 90%, 99%, or a range including and / or extending to the aforementioned values. In some embodiments, the treatment method described herein up-regulates the α-secretase activity of a subject. In some embodiments, the α-secretase activity increases by only or at least approximately those values equal to 150%, 170%, 180%, 190%, 199%, or a range including and / or extending to the aforementioned values. In some embodiments, the treatment method described herein up-regulates the neprilysin activity of a subject. In some embodiments, the neprilysin activity increases by only or at least approximately those values equal to 150%, 170%, 180%, 190%, 199%, or a range including and / or extending to the aforementioned values.

[0092] In some embodiments, administration of the compositions described herein can extend the lifespan, survival time, life duration, or health period of a subject. In some embodiments, the expected lifespan, survival time, life duration, or health period of the subject is the central expectation value for subjects in similar circumstances. In other embodiments, the expected lifespan, survival time, life duration, or health period of the subject is the average expectation value for subjects in similar circumstances. Subjects in similar circumstances can be determined based on any one or more factors including, but not limited to, age, health status, family history, or activity level.In some embodiments, the expected increase measured from the time treatment is initiated is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 40%, 50%, 1000%, any of the foregoing percentages, or a range bounded by any of the foregoing percentages (e.g., about 1% - 30%, about 5% - 25%, about 5% - 20%, about 5% - 15%, or 1% - 30%, 5% - 25%, 5% - 20%, 5% - 15%), 1% - 100%, 1% - 90%, 1% - 80%, 1% - 70%, 1% - 60%, 1% - 50%, 1% - 40%, 1% - 30%, 1% - 20%, 1% - 10%, 10% - 100%, 10% - 90%, 10% - 80%, 10% - 70%, 10% - 70%, 10% - 60%, 10% - 50%, 10% - 40%, 10% - 30%, 10% - 20%, 20% - 100%, 20% - 90%, 20% - 80%, 20% - 70%, 20% - 60%, 20% - 50%, 20% - 40%, 20% - 30%, 30% - 100%, 30% - 90%, 30% - 80%, 30% - 70%, 30% - 60%, 30% - 50%, 30% - 40%, 40% - 100%, 40% - 90%, 40% - 80%, 40% - 70%, 40% - 60%, 40% - 50%, 50% - 100%, 50% - 90%, 50% - 80%, 50% - 70%, 50% - 60%, 60% - 100%, 60% - 90%, 60% - 80%, 60% - 70%, 70% - 100%, 70% - 90%, 70% - 80%, 80% - 100%, 80% - 90%, 90% - 1000%, any of the ranges of the foregoing percentages (e.g., about 10% - 70%, about 30% - about 60%, or about 50% - 70%) as compared to the expected lifespan, survival time, life duration, or healthy period of the subject, and can be.In some embodiments, the expected increase is on an annual basis and is 1 to 40 years, 1 to 19 years, 1 to 18 years, 1 to 17 years, 1 to 16 years, 1 to 15 years, 1 to 14 years, 1 to 13 years, 1 to 12 years, 1 to 11 years, 1 to 10 years, 1 to 9 years, 1 to 8 years, 1 to 7 years, 1 to 6 years, 1 to 5 years, 1 to 4 years, 1 to 3 years, 1 to 2 years, 1 year, at least the foregoing years (e.g., at least 1 to 10 years), or about the foregoing years (e.g., about 1 to 2 years or at least about 1 to 2 years) compared to the expected lifespan, survival time, life duration, or healthy period of the subject. In some embodiments, the expected increase is on a daily to monthly basis and is 1 day to 1 year, 1 day to 11 months, 1 day to 10 months, 1 day to 9 months, 1 day to 8 months, 1 day to 7 months, 1 day to 6 months, 1 day to 5 months, 1 day to 4 months, 1 day to 3 months, 1 day to 2 months, 1 day to 1 month, at least the foregoing daily to monthly range (e.g., at least 1 day to 11 months), or about the foregoing daily to monthly range (e.g., about 1 day to 6 months or at least about 1 day to 6 months) compared to the expected lifespan, survival time, life duration, or healthy period of the subject.

[0093] Some embodiments relate to treating a disease or condition associated with the brain of a subject. In some embodiments, the β-amyloid 42 / β-amyloid 40 ratio of the subject decreases. In some embodiments, the composition improves the brain glucose metabolism of the subject. In some embodiments, the composition improves the cerebral blood flow of the subject. In some embodiments, the composition prevents or substantially prevents the hyperphosphorylation of tau protein. In some embodiments, the composition reduces β-amyloid in the brain of the subject. In some embodiments, the composition reduces the risk of cardiac arrest and stroke of the subject.

[0094] Some embodiments relate to increasing the mitochondrial activity of a subject's brain. In some embodiments, the mitochondrial activity of the subject's brain can increase by at least about 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or by a range that includes and / or extends between the aforementioned values. In some embodiments, the mitochondrial activity of the subject's brain can be restored. In some embodiments, the mitochondrial activity of the subject's brain is restored to at least 70% of normal activity, at least 75% of normal activity, at least 80% of normal activity, at least 85% of normal activity, at least 90% of normal activity, at least 95% of normal activity, or to a range that includes and / or extends between the aforementioned values.

[0095] In some embodiments, a method of treating cognitive impairment induced by amyloid-β protein according to the present disclosure can include administering a patch to a mammal (human or non-human mammal) or a model animal thereof that has developed cognitive impairment induced by amyloid-β protein. In some embodiments, a method of treating Alzheimer's disease according to the present disclosure can include administering a patch as described herein to a mammal (excluding humans) that has developed Alzheimer's disease or a model animal thereof.

[0096] Cognitive impairment induced by amyloid-β protein treated with a therapeutic agent according to the present disclosure typically refers to symptoms associated with Alzheimer's disease, but may also be symptoms associated with a disease that has not been definitively diagnosed as Alzheimer's disease, or may be symptoms in a preclinical stage subject or model animal including mild cognitive impairment (MCI). Model animals of cognitive impairment induced by amyloid-β protein can be prepared using known methods. For example, transgenic mice that exhibit overexpression of amyloid-β protein or mice administered a solution of amyloid-β protein in artificial cerebrospinal fluid can be used as such model animals.

[0097] In some embodiments, as discussed elsewhere herein, the subject is a human. However, the method is not limited to the treatment of humans and is equally applicable to the treatment of mammals. In such examples of treating non-human mammals, the selection of the patient depends on several factors within the skill of a veterinarian or research scientist who is a person of ordinary skill in the art.

[0098] System / Device The basic configuration of a micro-poration drug delivery system is known to those of ordinary skill in the art and thus need not be described in further detail herein. For example, a transdermal permeation delivery system is described in U.S. Patent No. 8,116,860, which is incorporated herein by reference for the purpose of illustrating various features of such a micro-poration drug delivery system in particular. As described therein, the micro-poration drug delivery system of U.S. Patent No. 8,116,860 (referred to therein using reference numeral "10") includes a filament array (referred to therein using reference numeral "70") configured to create one or more pathways or micropores in a patient's skin, and one or more transdermal patches (referred to therein using reference numeral "100") containing at least one drug formulation. Other similar micro-poration drug delivery systems including such basic features are known to those of ordinary skill in the art. Various micro-poration drug delivery systems having such basic features are known to those of ordinary skill in the art and may be used or adapted for use by those of ordinary skill in the art guided by the teachings provided herein.

[0099] In some embodiments, the micro - poration device can be defined by the total area of the paths (e.g., micropores) generated in the skin by one or more filaments and the total energy delivered to the one or more filaments to generate the paths. In some embodiments, the micro - poration device generates paths such that the total area of the paths in the skin is approximately 0.25 to 4.0 square centimeters (cm) of the skin. In some embodiments, the paths in the skin can preferably include approximately 0.5 to 12.5% of the skin area per square centimeter of the skin exposed to the micro - poration device. In some embodiments, the paths in the skin can preferably include approximately 1.25 to 10% of the skin area per square centimeter of the skin exposed to the micro - poration device.

[0100] In some embodiments, the energy delivered to the one or more filaments to generate the paths is 0.0067 μJ / μm 3 ~0.0400 μJ / μm 3can be in the range of. The energy delivered to one or more filaments can be delivered in pulses of 2 to 12 milliseconds (ms) for sufficient energy to create a consistent path of delivery. For example, the characteristics of the path for efficiently and safely delivering a drug through the skin from the patch described herein can vary between in vitro and in vivo embodiments. For example, in some embodiments, when energy of 2 mJ / filament to 12 mJ / filament is applied to one or more filaments during a pulse of 2 to 16 ms, one or more filaments create a path. In some embodiments, the energy applied to one or more filaments to create a path is 2 mJ / filament to 8 mJ / filament, 2 mJ / filament to 6 mJ / filament, or 2 mJ / filament to 4 mJ / filament. In some embodiments, the pulse duration is 2 to 12 ms. In such embodiments, one or more filaments can comprise stainless steel or can be substantially formed from stainless steel, and for 2 mJ / filament to 12 mJ / filament, 300,000 μm 3 (0.0067 μJ / μm 3 to 0.0400 mJ / μm 3 ) can have a volume (V).

[0101] In some embodiments, one or more filaments disposed in a filament array can create 25 to 500 paths per 1 cm 2 of the biological membrane (e.g., skin) to which one or more filaments are exposed. In some embodiments, one or more filaments disposed in a filament array can create 50 to 400 paths per 1 cm 2 of the skin to which one or more filaments are exposed.

[0102] In some embodiments, the cumulative (or total) depth of all paths formed by one or more filaments is approximately 2500 - 30000 μm per square centimeter of skin exposed to the one or more filaments. In some embodiments, the cumulative or total volume of all paths formed by one or more filaments is approximately 0.05 - 0.35 mm per square centimeter of skin. 3 is.

[0103] In some embodiments, the patch described herein may have one or more features that enhance the optimal drug release and diffusion of non-aggregated peptides into the body in conjunction with the paths generated by the micro-poration device.

[0104] In some embodiments, improvements to the micro-poration device and patch described herein enable the micro-poration drug delivery system to effectively and safely deliver non-aggregated peptides in a manner that provides improved bioavailability of the non-aggregated peptides and / or mobility from the subject's blood to the cerebrospinal fluid. Embodiments of the micro-poration drug delivery system described herein may provide improved patient compliance and enhanced drug delivery capabilities. Embodiments of the micro-poration drug delivery system may also reduce the risk of adverse effects caused by uncontrolled delivery and reduce the development time and cost of drugs for patients. Embodiments of the micro-poration drug delivery system also enable the patient to painlessly and needlelessly self-administer the corresponding drug at the location chosen by the patient, resulting in improved compliance and cost reduction (reduction in visits to healthcare providers). Embodiments of the micro-poration drug delivery system described herein have less variability in individual drug delivery results and can be used in patients with a wide range of skin types, conditions, etc.

[0105] Examples Various embodiments and alternatives are disclosed in more detail in the following examples, which are in no way intended to limit the claims.

[0106] Alzheimer's disease. Alzheimer's disease is a chronic disease that progresses with aging. Therefore, due to the nature of Alzheimer's disease, medication may require daily administration over a long period. Thus, intravenous administration is not desirable as it requires hospitalization or daily visits. Subcutaneous administration can be developed as a self-administration device such as a pen-type syringe. However, due to patient pain and handling issues, it is considered inefficient for most patients over the long term. Furthermore, patients may have needle phobia. Therefore, transdermal micro-poration is thought to have good compliance and function as long-term home treatment. In this study, to enhance the delivery of p3-Alcβ to the brain and determine the effectiveness of various administration modes, a transdermal micro-poration device was investigated for the non-aggregating peptide p3-Alcβ in comparison with other administration modes.

[0107] p3-Alcβ. Figure 1 depicts the amino acid sequences of p3-Alcβ1-40, p3-Alcβ1-37, p3-Alcβ9-19, p3-Alcβ1-19, p3-Alcβ11-19, which are partial peptides of arcadien β. A human p3-Alcβ37 peptide containing the sequence of Val813 - Thr849 of Alcβ and its partial peptide p3-Alcβ9-19 described in Hata et al., "Alcadein cleavages by amyloid β-precursor protein (APP) α- and γ-secretase generates small peptides, p3-Alcs, indicating Alzheimer’s disease-related γ-secretase dysfunction" J. Biol. Chem.

[2009] 284, 36024 - 36033 was synthesized and purified to a purity of over 95%. Their predicted molecular weights were confirmed by mass spectrometry performed at Peptide Institute (Osaka, Japan).

[0108] Animal tests in rats. Oral, nasal, injection, and transdermal micro-poration containing p3-Alcβ9-19 were tested to determine pharmacokinetics such as PK profiles, bioavailability, and drug recovery rates in vivo. In vivo PK tests were performed in CD rats (Charles River Laboratories) according to the IACUC-approved animal protocol. Each dosage formulation was prepared before the experiment. The patches were manufactured according to the dry patch manufacturing procedure. Animals for transdermal application were shaved one day before the administration date. Transdermal micro-poration was applied using an applicator and a porator. Then, the patch was placed to cover the poration area for one day or the designated period. After the last blood collection, the patch was recovered, the application site was wiped with paper or cotton, and the residual drug on the skin was recovered. At the designated sampling times, blood samples were collected from the tail vein and transferred to K2-EDTA tubes. The samples were centrifuged at 4°C and then plasma samples were obtained. All samples were stored in a -80°C freezer until bioassay.

[0109] Sample preparation for LCMSMS analysis of p3-Alcβ9-19. Plasma samples were placed at room temperature for 30 minutes. Plasma (50 μL) was transferred to a 2 mL Eppendorf tube (Protein LoBind), then 50 μL of acetonitrile / DMSO (4:1, v / v) was added, and the tube was vortexed for several seconds. The sample was centrifuged at 3700 rpm at 4°C for 15 minutes. 150 μL of mobile phase A (0.1% formic acid in water) was placed in a new tube, then 50 μL of the supernatant was transferred to the tube, and the sample was vortexed for 5 minutes. The supernatant was taken out, and the solution was filtered through a 0.20 μm PTFE membrane into an autosampler vial (LCMSMS).

[0110] LCMSMS method for p3-Alcβ9-19. The LCMSMS conditions are shown in Table 1 and Table 2.

Table 1

Table 2

[0111] Animal tests in mice. The transport of p3-Alcβ9-19 into the blood and cerebrospinal fluid (CSF) was tested in wild-type mice (4 months old). The hair was removed before the experiment. Percutaneous micro-poration was applied, and then a patch containing p3-Alcβ9-19 was administered to the porated area. The mice were anesthetized by using 1% isoflurane, and CSF was collected from the cisterna magna of the mice into a tube containing EDTA and heparin, and blood was collected from the inferior vena cava as described in Liu et al., "A technique for serial collection of cerebrospinal fluid from the cisterna magna in mouse." J.Vis.Exp.21(2008).

[0112] Development of an sELISA method for p3-Alcβ9-19 analysis. An sELISA method was developed to measure p3-Alcβ9-19 in cerebrospinal fluid (CSF). A polyclonal rabbit antibody against p3-Alcβ9-19 was raised against p3-Alcβ9-19 containing an amino-terminal Cys residue (C+HRGHQPPPEMA) and conjugated to bovine serum globulin. The IgG was purified with antigen-binding resin and conjugated to biotin. The horseradish peroxidase-conjugated streptavidin was manufactured by Amersham / GE Healthcare (Little Chalfont, UK, catalog number RPN1051), and the tetramethylbenzidine (TMB) micro-well peroxidase substrate system was manufactured by SeraCare Life Sciences Inc. (Milford, Massachusetts, USA, catalog number 5120-0075). The mice were anesthetized using 1% isoflurane, CSF was collected from the cisterna magna of the mice as described above, and then the mice were sacrificed.

[0113] To quantify p3-Alcβ9-19 in mouse CSF and plasma, samples were diluted with buffer A (PBS containing 1% bovine serum albumin and 0.05% Tween-20). Using a polyclonal antibody, the developed sELISA system quantified p3-Alcβ9-19 in the range of 25-200 pg / ml, with sensitivity equivalent to that of the p3-Alcα35-specific sELISA system described by Omori et al., "Increased levels of plasma p3-Alcα35, a major fragment of Alcadeiνα by γ-secretase cleavage, in Alzheimer’s disease" J. Alzheimers Dis. (2014) 39, 861-870, and the p3-Alcβ37 and p3-Alcβ40-specific sELISA systems described by Hata et al., "Decrease in p3-Alcβ37 and p3-Alcβ40, products of Alcadeinβ generated by γ-secretase cleavages, in aged monkeys and patients with Alzheimer’s disease" Alzheimers Dement TRCI (2019) 5, 740-750. In this p3-Alcβ9-19 sELISA system, antiserum diluted 1:10,000 was used as the capture antibody, and affinity-purified antiserum with antigen-binding resin and biotin-labeled IgG was used as the detection antibody. This sELISA system did not react with p3-Alcβ37. The addition of 1,000 pg / ml of p3-Alcβ37 did not compete with antibody binding to p3-Alcβ9-19 in the range of 0-200 pg / ml. The sELISA method was used to measure p3-Alcβ9-19 in body fluids even in the presence of endogenous p3-Alcβ in mice.

[0114] sELISA method for p3-Alcβ9-19 analysis. Rabbit antiserum immunized with p3-Alcβ9-19 as an antigen was coated on a 96-well plate at 4 °C overnight as a capture antibody. The next day, unbound antibodies were removed with washing buffer (0.05% Tween-20 in PBS). Plasma and CSF were diluted 200-fold and 20-fold, respectively, with EIA buffer (1% BSA, 0.05% Tween-20). The sample solution was added to the 96-well plate and incubated at 4 °C overnight. The next day, after washing with washing buffer, purified biotinylated IgG dissolved in PBS was added to each well to 1.0 μg / mL, and the mixture was maintained at 4 °C overnight. The next day, unreacted antibodies were washed with washing buffer, streptavidin-HRP (1 / 5000 in PBS) was added, and the reaction was carried out at 4 °C for 6 - 8 hours. After washing with washing buffer, TMB chromogenic substrate was added, and the mixture was reacted at room temperature for 30 minutes in the dark. Color development was stopped by adding 1N H2SO4, and the samples were measured at 450 nm with a plate reader.

[0115] Intracerebral mitochondrial activation. The increase in neuron survival rate by p3-Alcβ was verified in an in vivo setting by monitoring brain mitochondrial function using [18F]BCPP-EF probe-based PET imaging, which can detect mitochondrial complex I activity reflecting neuron survival rate in the surviving brain. Mitochondrial dysfunction generally exists in the brains of AD patients, and the decrease in survival rate in vulnerable brain regions can be detected by PET imaging using [18F]BCPP-EF.

[0116] Transdermal microporation device. FIG. 2A depicts an example of a patch, and FIG. 2B shows patch application after transdermal microporation in the skin. The microporation device comprises (a) an applicator electrically connected to filaments, (b) a porator having an upper substrate surface and defining a poration region, and (c) a patch. A filament array having a plurality of filaments is disposed in the poration region, and each filament can conductively deliver thermal energy through direct contact with the tissue membrane to form at least one micropore in the tissue membrane. The applicator supplies a predetermined electrical energy to the filaments to generate micropores, and the patch is applied to at least a part of the micropores. The patch (100) comprises a backing (200), a matrix (300) containing a composition (302) such as p3-Alcβ and components in a dry state, and a release liner (400) that is removed before application to the porated skin. The backing (200) may have an adhesive (201). The matrix (300) can consist of a matrix support (301) and a composition (302) disposed on the matrix support (301). After transdermal microporation, the patch (100) is applied to the porated skin after removing the release liner (400). The active ingredient in the composition (302) is dissolved by the interstitial fluid from the microchannels, and then the active ingredient moves to the microchannels (transdermal region) and then enters the blood circulation.

[0117] Patch manufacturing. Patches were manufactured based on dispensing or tableting procedures known to those skilled in the art. Individual formulations utilized the different components described herein. The patch includes a backing, a matrix containing a non-aggregated peptide and components disposed within the matrix, and a release liner, which is configured to be removed prior to application to the skin of a subject. The non-aggregated peptide is p3-Alcβ9-19. In the dispensing method, all components were dissolved in a mixed solvent of water and ethanol. A predetermined amount of the component solvent was dispensed onto the non-woven pad in the patch without the release liner. The dispensed patch was transferred to a drying chamber at 50 °C until dried. The dried patch was removed from the drying chamber and placed at room temperature for 30 minutes. The patch was covered with the release liner and then cut to the specified size. Individual patches were packed into an aluminum laminate pouch together with a desiccant. In the tableting method, all components were mixed and weighed as a specified amount. The weighed powder was transferred to a die punch of a specified size for the press. The die was set on the machine and compressed. The thin rectangular tablets were taken out and placed on the non-woven pad in the patch without the release liner. The patch was covered with the release liner and then cut to the specified size. Individual patches were packed into an aluminum laminate pouch together with a desiccant.

[0118] Example 1. Administration Route of p3-Alcβ Non-parenteral administration routes. p3-Alcβ9-19 (5 mg / head) was administered to rats via oral, nasal, or passive transdermal routes. No absorption was observed in all administrations. p3-Alcβ9-19 is a hydrophilic peptide and its molecular weight is 1256 daltons. These physicochemical properties can affect the poor membrane permeability through the gastrointestinal tract, nose, and stratum corneum.

[0119] Parenteral administration route. Figures 3A, 3B, and 3C show the PK profiles (mean ± SE, n = 4) of p3-Alcβ9-19 after intravenous and subcutaneous administrations in rats. Figure 3A shows a line graph depicting the PK profile of p3-Alcβ9-19 after intravenous administration (1 and 2 mg / body) in rats, Figure 3B shows a line graph depicting the PK profile of p3-Alcβ9-19 after subcutaneous administration (1 - 10 mg / body), and Figure 3C shows a logarithmic scale graph depicting the PK profile of p3-Alcβ9-19 on a logarithmic scale. Figure 4A shows a line graph depicting the dose response of p3-Alcβ9-19 after intravenous and subcutaneous administrations in rats, and Figure 4B shows a line graph depicting the dose response of p3-Alcβ9-19 after subcutaneous administration in rats (mean ± SE, n = 4).

[0120] The PK profiles of intravenous (IV) injection and subcutaneous (SC) injection showed rapid absorption and rapid elimination in rats. The blood concentration in SC persisted slightly longer than that in IV but disappeared within 1 hour. A dose-dependent increase was observed in both IV injection and SC injection. The bioavailability in SC can be the same as or slightly higher than that in IV injection.

[0121] Delivery of p3-Alcβ using transdermal micro-poration in rats. Figure 5A shows a line graph depicting an immediate release (IR) formulation. Figure 5B shows a line graph depicting the dose response from transdermal micro-poration (400 density, 4 mJ / filament). Tables 3 and 4 describe the formulations and results of this study, respectively.

Table 3

Table 4

[0122] Transdermal microporation-mediated delivery of p3-Alcβ9-19 showed 545 - 889% higher aBA compared to IV (2 mg / body). A dose-dependent increase was observed with the transdermal microporation system. Microporation-mediated delivery was the highest even when compared to IV injection and SC injection. The parenteral route may be affected by the in vivo stability of p3-Alcβ9-19, and it was suggested that microporation-mediated delivery could avoid the first-pass effect (metabolism), and the sustained release of p3-Alcβ9-19 rather than injection showed higher bioavailability.

[0123] Example 2. Brain delivery of p3-Alcβ using transdermal microporation in mice The formulations used in the tests are shown in Table 5. Figure 6A shows a logarithmic scale graph depicting the changes in blood concentration and central concentration after an IR formulation containing p3-Alcβ9-19 using transdermal microporation in mice. Figure 6B shows a logarithmic scale graph depicting the changes in blood concentration and central concentration after a sustained release (SR) formulation containing p3-Alcβ9-19 using transdermal microporation in mice. Table 6 describes the PK parameters in plasma after administration of p3-Alcβ9-19 in mice. [Table 5] [Table 6]

[0124] Table 7 describes the PK parameters in cerebrospinal fluid (CSF) after administration of p3-Alcβ9-19 in mice. [Table 7]

[0125] Figures 6A and 6B show the changes in the plasma and cerebrospinal fluid (CSF) concentrations of p3-Alcβ9-19 when an IR formulation (Figure 5A) or an SR formulation (Figure 5B) containing p3-Alcβ9-19 was administered to mice using a transdermal microporation device. Tables 2 and 3 show that when an IR formulation containing 1 mg of p3-Alcβ9-19 was administered to mice, the maximum plasma concentration (Cmax) was approximately 600 ng / mL at 1 hour after administration, and the plasma concentration was maintained at approximately 300 ng / mL for a maximum of 6 hours. In the CSF, more than 20 ng / mL was shown 3 hours after administration, and a concentration of approximately 5 ng / mL was detected 6 hours after administration. The transfer rate of p3-Alcβ9-19 from blood to CSF being approximately 3.5% (BA in CSF / BA in plasma × 100%) indicated good central transfer. The SF formulation showed a sustained plasma concentration in mice, and the CSF concentration was close to the detection limit.

[0126] Example 3. Pharmacological Effects of Transdermal Microporation of p3-Alcβ in Monkeys In this example, two patch formulations of the present disclosure are exemplified. It was suggested that the use of an IR patch is preferably considered to achieve an effective increase in mitochondrial activity.

[0127] The mitochondrial activation effect of p3-Alcβ9-19 transdermal microporation was investigated in rhesus monkeys. The formulations used in the test are shown in Table 8 below. As a result, 1.5 hours after the administration of p3-Alcβ9-19 using transdermal microporation, [18F]BCPP-EF accumulation, that is, mitochondrial activation, was observed by PET measurement. Furthermore, activation continued even 6.5 hours later in some regions.

Table 8

[0128] Figure 7 shows two images depicting the changes in mitochondrial activity in the brain after the administration of an IR formulation (left) and an SR formulation (right) containing p3-Alcβ9-19 using transdermal micro-poration in monkeys. Transdermal micro-poration was applied at a density of 400 and 4 mJ / filament for the IR formulation, and at a density of 100 and 3 mJ / filament for the SR formulation. The increase in mitochondrial activity was observed in monkeys as an immediate response (early), 1 hour after the administration of the IR formulation containing p3-Alcβ9-19, and as a delayed response (delayed), 6.5 hours after the administration of the SR formulation containing p3-Alcβ9-19, relative to the control (vehicle) (marked by the red rectangles).

[0129] Figure 8 shows two bar graphs explaining the intensities from Figure 7. In Figure 8, the following abbreviations are used: fro: frontal lobe, tem: temporal lobe, par: parietal lobe, occ: occipital lobe, hipp: hippocampus, cd: caudate nucleus, put: putamen. Figure 8 (top) shows the administration of the IR formulation, and Figure 8 (bottom) shows the administration of the SR formulation. In Figure 8 (top), compared to the control (vehicle), the intensity increased at 1 hour in all regions, but decreased at 6.5 hours in all regions except the putamen. In Figure 8 (bottom), the intensity gradually increased in all regions.

[0130] Figure 9 shows a bar graph indicating the increase rate of mitochondrial activity in various regions of the brain 1 hour after the administration of an IR formulation containing p3-Alcβ9-19 (corresponding to 5 or 10 mg / body, equivalent to 0.5 or 1.0 mg / kg, respectively). The increase rate was higher at 10 mg / body than at 5 mg / body, except in the frontal lobe region and the caudate nucleus region.

[0131] Example 4. Control of PK Profile Using IR Formulation Table 9 describes the IR formulations used in the tests, which contain p3-Alcβ9-19, an enhancer (i.e., non-reducing sugar: sucrose), and an antimicrobial agent (a combination of methylparaben and propylparaben or benzalkonium chloride). To investigate PK modulation, transdermal micro-poration was applied in different combinations of density and energy levels.

Table 9

[0132] Figure 10A shows a line graph depicting the PK profiles using non-reducing sugar formulations and time-dependent p3-Alcβ concentrations with different densities at 4 mj / filament, and Figure 10B shows a logarithmic scale graph depicting the time-dependent p3-Alcβ concentration.

[0133] Table 10 shows the results of the type of PK profiles using IR formulations containing p3-Alcβ9-19 under different micro-poration conditions, including the results of Figure 10A. In immediate release (IR) type formulations, the PK profile can be controlled by changing the transdermal micro-poration conditions. Absorption was not observed without transdermal micro-poration. The 200 density at 4 mJ / filament and the 400 density at 2 - 4 mJ / filament showed the immediate PK profile of p3-Alcβ. On the other hand, the micro-poration conditions (50 ≤ density ≤ 200 density and < 4 mJ / filament) provided a sustained PK profile of p3-Alcβ.

Table 10

[0134] Example 5. Control of PK Profile Using SR Formulation Table 11 describes the SR formulation used in the test, which contains p3-Alcβ9-19, an enhancer (i.e., non-reducing sugar: sucrose), a PK regulator (disodium citrate sesquihydrate), and an antimicrobial agent (methylparaben / propylparaben).

Table 11

[0135] Table 12 provides an overview of the types of PK profiles using SR formulations containing p3-Alcβ9-19 with different contents of PK regulators. The PK regulator (i.e., containing an organic acid, its salt, or a combination thereof) is added in a sustained release (SR) type formulation. A sustained PK profile of p3-Alcβ9-19 was obtained with 4 mg of disodium citrate sesquihydrate at a density of 200 and 4 mJ / filament.

Table 12

[0136] Table 13 describes that the SR formulation used in the test contained p3-Alcβ9-19, an enhancer (non-reducing sugar: sucrose), a PK regulator (disodium citrate sesquihydrate), and an antimicrobial agent (methylparaben / propylparaben). Percutaneous micro-poration was applied at different combinations of density and energy levels using the PK regulator (4 mg of disodium citrate sesquihydrate).

Table 13

[0137] Table 14 summarizes the PK profiles using different micro-poration conditions with a PK regulator. The sustained release (SR) formulation containing disodium citrate sesquihydrate showed a sustained PK profile with 4 mg of disodium citrate sesquihydrate at a density less than 400 and 4 mJ / filament.

Table 14

[0138] Table 15 summarizes the PK profile control using IR formulation and SR formulation containing p3-Alcβ with transdermal micro-poration. The immediate PK profile is provided by IR type formulation (without PK regulator) under micro-poration conditions of less than 200 density and 2 - 4 mJ / filament. The sustained PK profile can be obtained with IR formulation of 50 ≤ density ≤ 200 density and < 4 mJ / filament, and SR formulation containing PK regulator at less than 400 density and 4 mJ / filament.

Table 15

[0139] Example 6. Enhancement of p3-Alcβ delivery by enhancer Table 16 describes a dry patch formulation for investigating the effect of enhancer (sugar) on the delivery of p3-Alcβ9-19 using transdermal micro-poration. The enhancers were selected from non-reducing sugars (sucrose, trehalose, D-mannitol, and D-sorbitol) and reducing sugars (lactose and maltose).

Table 16

[0140] Figure 11 shows the PK profiles by non-reducing sugars or reducing sugars. Reducing sugars (lactose and maltose) showed higher delivery than non-reducing sugars. Table 17 describes the results from Figure 11. All non-reducing sugars or reducing sugars enhanced p3-Alcβ absorption compared to IV injection. The aBA of non-reducing sugars was 194.3 - 547.3%. In particular, reducing sugars such as maltose and lactose showed significant enhancement for p3-Alcβ9-19 using transdermal micro-poration (aBA: 2923.3 and 4121.7%).

Table 17

[0141] Example 7. Effect of Antimicrobial Agents on p3-Alcβ Delivery Antimicrobial agents are added as preservatives or antimicrobial effectiveness to avoid the risk of infectious diseases. Table 18 describes examples of formulations containing antimicrobial agents. [Table 18]

[0142] Table 19 describes the PK parameters after administration of formulations using percutaneous micro-poration (400 density, 4 mJ / filament) in rats. It is suggested that reducing sugars act together with antimicrobial agents (formulations of combinations of methylparaben and propylparaben, or benzalkonium chloride). [Table 19]

[0143] Example 8. Immediate and Sustained PK Delivery Using Reducing Sugar Formulations Table 20 describes IR (without PK regulator) formulations and SR (containing PK regulator) formulations containing reducing sugar (lactose) and p3-Alcβ9-19 for controlling the PK profile. [Table 20]

[0144] Figure 12A shows an example of a line graph showing the PK profiles of IR formulations and SR formulations containing reducing sugar (lactose) and p3-Alcβ formulations in rats (G2 and G4), and Figure 12B shows a logarithmic scale graph showing the p3-Alcβ concentration over time. After administration of IR formulations and SR formulations containing reducing sugar using percutaneous micro-poration, the plasma concentration of p3-Alcβ9-19 showed typical immediate and sustained PK profiles.

[0145] Table 21 describes the results of PK parameters after administration of formulations using percutaneous micro-poration in rats. The IR formulation and SR formulation containing reducing sugar showed high BA compared with the non-reducing sugar formulation, respectively.

Table 21

[0146] Example 9. Effect of density on p3-Alcβ delivery using reducing sugar formulations Table 22 describes the formulations for determining p3-Alcβ 9-19 delivery at different micro-poration densities.

Table 22

[0147] Figure 13 shows the drug residue at 24 hours after administration of the reducing sugar formulation using percutaneous micro-poration (4 mJ / filament) in rats. p3-Alcβ 9-19 was recovered at about 100% without micro-poration. The drug recovery rate decreased as the percutaneous micro-poration density increased. 10% - 20% of p3-Alcβ 9-19 was recovered at densities of 200 - 400. Above 200 density indicates effective delivery of p3-Alcβ 9-19.

[0148] Example 10. Optimization of enhancer content in reducing sugar formulations Table 23 describes the formulations used to investigate the effect of enhancer content in p3-Alcβ percutaneous micro-poration.

Table 23

[0149] Figure 14 shows a line graph indicating the PK profiles of rats at different enhancer contents (reducing sugar, lactose). Table 24 describes the PK parameters from Figure 14. Figure 15 shows a line graph comparing AUC against lactose content.

Table 24

[0150] The plasma concentration of p3-Alcβ9-19 increased with the increase in enhancer content (G1 - G5). No enhancement was observed without the enhancer (G7). The ratio of p3-Alcβ to the enhancer may be related to the augmentation factor.

[0151] Example 11. Dose Dependency of p3-Alcβ Delivery Using IR and SR Formulations Immediate delivery system. Tables 25 and 26 describe IR formulations containing p3-Alcβ for determining dose dependency using transdermal micro-poration. [Table 25] [Table 26]

[0152] Table 27 shows the PK parameters in immediate delivery at different p3-Alcβ9-19 doses using transdermal micro-poration in rats (400 density, 4 mJ / filament). [Table 27]

[0153] Figure 16 shows the relationship between p3-Alcβ9-19 dose and AUC. Linear dose dependency in immediate delivery was observed up to 50 mg of p3-Alcβ9-19 (test dose).

[0154] Sustained delivery system. Tables 28 and 29 describe SR formulations containing p3-Alcβ for determining dose dependency using transdermal micro-poration. [Table 28] [Table 29]

[0155] Table 30 shows the PK parameters in sustained delivery at different p3-Alcβ9-19 doses using transdermal micro-poration in rats (100 density, 3 mJ / filament). [Table 30]

[0156] Figure 17 shows the relationship between the p3-Alcβ9-19 dose and the AUC. Linear dose-dependency in sustained delivery was observed with up to 25 mg of p3-Alcβ9-19 (test dose).

[0157] Example 13. Optimized patch formulation for p3-Alcβ delivery Tables 31 and 32 describe the optimized IR formulations and patch materials for immediate delivery. [Table 31] [Table 32]

[0158] Figure 18 shows a line graph depicting the PK profile of a formulation containing p3-Alcβ9-19 optimized for immediate delivery in rats. Table 33 describes the results of Figure 18. p3-Alcβ9-19 was efficiently delivered from the patch, and an aBA of approximately 2000% relative to IV injection was obtained. Tmax was approximately 1 hour. [Table 33]

[0159] Figure 19A depicts the relationship between the p3-Alcβ9-19 dose and the AUC. Figure 19B depicts the relationship between the p3-Alcβ9-19 dose and the Cmax. The AUC and Cmax increased in a p3-Alcβ9-19 dose-dependent manner.

[0160] Physicochemical properties of the p3-Alcβ9-19 patch. The pH and water content of the formulation were tested. Tables 34 to 36 describe the results. The pH was measured after mixing the patch with water (USP, pH 5 - 7). The pH of the formulation was approximately 4.9 (the acceptable range of pH was set at 4 - 9). The water content of the patch was measured by a volumetric Karl Fischer apparatus after mixing the patch with dehydrated methanol. The initial water content was less than approximately 2%. Dissolution was tested using Apparatus 5 (paddle and disk). The patch was placed in 500 mL of phosphate buffered saline (USP) at 50 rpm and 32 °C. The drug was measured by RP-HPLC. The drug was released immediately within 5 - 10 minutes. [Table 34] [Table 35] [Table 36]

[0161] Preliminary stability of the p3-Alcβ9-19 patch. The preliminary stability of the formulation was evaluated at 25 °C / 60% RH and 40 °C / 75% RH for 3 months. Tables 37 to 39 describe the results of the p3-Alcβ9-19 (5 mg) patch, p3-Alcβ9-19 (20 mg) patch, and p3-Alcβ9-19 (50 mg) patch, respectively. All formulations were stable under the test conditions. [Table 37] [Table 38] [Table 39]

[0162] Table 40 summarizes the findings of the p3-Alcβ administration route. Transdermal micro-poration for delivering p3-Alcβ9-19 is an ideal administration device, and the patch formulation shows good stability against storage at room temperature. This is important for patient compliance and the supply chain.

Table 40

[0163] Furthermore, although the above has been described in some detail by way of illustration and example for clarity and understanding, it will be understood by those skilled in the art that numerous various modifications can be made without departing from the spirit of the present disclosure. Accordingly, the forms disclosed herein are merely illustrative and are not intended to limit the scope of the present disclosure. Rather, it should be clearly understood that all modifications and alternative forms that accompany the true scope and spirit of the present disclosure are also included.

Claims

1. A patch for delivering a non-aggregating peptide, comprising: a backing; a matrix containing the non-aggregating peptide disposed within the matrix; and a release liner wherein the release liner is configured to be removed prior to application to the skin of a subject.

2. The patch according to claim 1, wherein the non-aggregating peptide is p3-Alcβ.

3. The patch according to claim 2, wherein the p3-Alcβ is selected from at least one of p3-Alcβ1-40, p3-Alcβ1-37, p3-Alcβ9-19, p3-Alcβ1-19, p3-Alcβ11-19, or a derivative thereof.

4. The patch according to claim 2, wherein the p3-Alcβ is p3-Alcβ9-19 or a derivative thereof.

5. The non-aggregating peptide is present in the matrix in an amount in the range of about 0.01 mg / cm ２ to about 200 mg / cm ２ of the patch according to any one of claims 1 to 4.

6. The patch according to any one of claims 1 to 5, wherein the matrix further comprises at least one sugar.

7. The patch according to claim 6, wherein the at least one sugar is a non-reducing sugar, a reducing sugar, or a combination thereof.

8. The patch according to claim 7, wherein the non-reducing sugar is sucrose, trehalose, mannitol, sorbitol, or a combination thereof.

9. The patch according to claim 7, wherein the reducing sugar is lactose, maltose, or a combination thereof.

10. The patch according to any one of claims 1 to 6, wherein the weight ratio of the at least one sugar to the non-aggregating peptide is greater than about 0.

02.

11. The patch according to any one of claims 1 to 6, wherein the weight ratio of the at least one sugar to the non-aggregating peptide is from about 0.02 to about 0.

4.

12. The patch according to any one of claims 1 to 11, wherein the matrix further comprises at least one drug delivery regulator.

13. The patch according to claim 12, wherein the at least one drug delivery regulator is an organic acid, a salt thereof, or a combination thereof.

14. The patch according to claim 13, wherein the at least one drug delivery regulator is citric acid, a salt form thereof, or a combination thereof.

15. The patch according to any one of claims 1 to 14, wherein the matrix further comprises a preservative.

16. The patch according to claim 15, wherein the preservative is an antimicrobial agent.

17. The patch according to claim 16, wherein the antimicrobial agent is selected from the group consisting of methylparaben, propylparaben, benzalkonium chloride, sodium benzoate, and combinations thereof.

18. The patch according to any one of claims 1 to 17, wherein the matrix further comprises at least one of sucrose, lactose, disodium citrate sesquihydrate, methylparaben, propylparaben, and benzalkonium chloride.

19. The patch according to any one of claims 1 to 18, wherein the matrix comprises at least one kind of fiber, a film lamination material, or a combination thereof.

20. The patch according to claim 19, wherein the matrix comprises a film lamination material and at least one kind of fiber.

21. The patch according to claim 19 or 20, wherein the at least one kind of fiber is a non-woven fiber.

22. The patch according to any one of claims 19 to 21, wherein the at least one kind of fiber has a thickness of less than about 300 μm.

23. The at least one fiber has a weight of less than about 100 g / m ２ The patch according to any one of claims 19 to 22, having a weight of less than ２ .

24. The matrix has a water retention capacity of less than about 10 mg / cm ２ The patch according to any one of claims 1 to 23, which has a water retention capacity of less than ２ .

25. A device for delivering a non-aggregated peptide through a plurality of microchannels, a patch according to any one of claims 1 to 24, a porator including an array of conductive filaments, and an applicator electrically connected to the conductive filaments and configured to supply a predetermined electrical energy to the array of conductive filaments to generate a plurality of micropores in a micropore region of the skin of the subject by heating the filaments. A device comprising.

26. from about 25 to about 500 microchannels / cm ２ The device according to claim 25, which generates ２ .

27. The device according to claim 25 or 26, having a poration energy of about 2 to about 6 mJ / filament, 4 mJ / filament, or 8 mJ / filament.

28. The porator is configured to open at least one channel in the skin of the subject, about 0.25 cm ２ to about 4 cm ２ in area, the device according to any one of claims 25 to 27.

29. The porator is configured to open at least one channel within the skin of the subject, about 0.1 cm ２ , about 0.25 cm ２ , about 0.65 cm ２ , or about 1.0 cm ２ in area, the device according to claim 28.

30. The porator is configured to open at least one channel in the skin of the subject, about 1.0 cm ２ less than, about 0.5 cm ２ less than, or about 0.25 cm ２ The device according to claim 28 or 29, having an area less than.

31. The device according to claim 30, wherein the at least one channel is one or more micropores, and the porator is configured to generate the one or more micropores in about 0.5% to about 12.5% of the total poration region.

32. The device according to any one of claims 25 to 31, wherein the porator is a micro-needle, a laser, or a high-frequency porator configured to generate one or more micropores in the skin of the subject.

33. The device according to any one of claims 25 to 32, wherein the porator is configured to generate at least 50 pores in the target skin.

34. The device according to any one of claims 25 to 33, wherein the porator is a microneedle, a laser, or a radiofrequency porator.

35. The device according to any one of claims 25 to 34, wherein the patch further comprises a drug pellet.

36. A method of treating a disease or condition associated with the brain of a subject, comprising: opening at least one channel in the skin of the subject; and applying a patch according to any one of claims 1 to 24 to the skin of the subject. A method comprising the steps of:

37. The method according to claim 36, wherein the disease or condition associated with the brain of the subject is a neurodegenerative disease.

38. The method according to claim 37, wherein the neurodegenerative disease is Alzheimer's disease.

39. The method according to any one of claims 36 to 38, wherein the step of opening at least one channel in the skin of the subject comprises applying a transdermal microporation device to the skin of the subject.

40. The method according to claim 39, wherein the transdermal microporation device is thermal tissue ablation by using a filament array having a plurality of filaments disposed on the skin of the subject, and each filament can conductively deliver thermal energy through direct contact with the tissue membrane to form the plurality of micropores in the micropore region of the tissue membrane.

41. The transdermal microporation device generates about 25 to about 500 microchannels / cm ２ The method according to claim 36 or 40, wherein the method is performed.

42. The method according to any one of claims 39 to 41, wherein the transdermal microporation device has a poration energy of about 2 to about 6 mJ / filament, about 4 mJ / filament, or about 8 mJ / filament.

43. The step of opening at least one channel in the skin of the subject is from about 0.25 cm ２ to about 4 cm ２ The method according to any one of claims 36 to 42, having an area of.

44. The method according to any one of claims 36 to 38, wherein the transdermal microporation device is a microneedle, a laser, or a radiofrequency device capable of generating one or more micropores in the skin of the subject.

45. The method according to any one of claims 36 to 44, wherein administering the patch increases the neuronal survival rate in the subject.

46. The method according to any one of claims 36 to 44, wherein mitochondrial activity in the brain of the subject is increased by applying the patch.

47. The method according to any one of claims 36 to 46, wherein the patch provides a maximum blood concentration of the non-aggregated peptide at least 0.5 hours after administration.

48. The method according to any one of claims 36 to 46, wherein the non-aggregated peptide is maintained for at least 6 hours after administration of the patch.

49. The method according to any one of claims 36 to 46, wherein the non-aggregated peptide from the patch is maintained in the blood of the subject for at least 6 hours after administering the patch to the subject.

50. The method according to any one of claims 36 to 49, wherein the migration rate of the non-aggregated peptide from the blood of the subject to the cerebrospinal fluid of the subject is at least 2%.

51. The step of opening at least one channel in the skin of the subject is about 0.1 cm ２ , about 0.25 cm ２ , about 0.65 cm ２ , or about 1.0 cm ２ in area, the method according to claim 43.

52. A transdermal delivery system for delivering a non-aggregated peptide to a subject, a substrate having an upper substrate surface and defining a poration region, the substrate including a filament array having a plurality of filaments disposed in the poration region, each filament capable of conductively delivering thermal energy through direct contact with a tissue membrane to form a plurality of micropores in a micropore region of the skin; an applicator electrically connected to the filament array and configured to supply a predetermined electrical energy to the filaments to generate the plurality of micropores in the micropore region of the skin by heating the filaments; a power circuit configured to supply a current to the applicator; a patch according to any one of claims 1 to 24 and a system comprising the same.

53. The system according to claim 52, wherein the patch is for application to the one or more micropores.

54. The transdermal micro-poration device generates about 25 to about 500 micro-channels / cm ２ The system according to claim 52 or 53, which generates the same.

55. The system according to claim 52 or 54, wherein the transdermal microporation device has a poration energy of about 2 to about 10 mJ / filament.

56. The step of opening at least one channel in the skin of the subject is about 0.25 cm ２ to about 4 cm ２ The system according to any one of claims 52 to 55, having an area of

57. The system according to any one of claims 52 to 56, wherein the one or more micropores are about 0.5 to about 12.5% of the total poration region.

58. The system of claim 52, wherein the transdermal microporation device is a microneedle, laser, or radiofrequency device capable of generating one or more micropores in the skin of the subject. **Claim 59** The system according to any one of claims 52 to 58, wherein the transdermal microporation device generates at least 50 pores in the skin of the subject. **Claim 60** The system according to any one of claims 52 to 59, wherein the patch further comprises a drug pellet. **Claim 61** The substrate is configured to open at least one channel within the skin of the subject, about 0.1 cm ２ , about 0.25 cm ２ , about 0.65 cm ２ , or about 1.0 cm ２ in area, the system of claim 56.

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