Transdermal delivery devices for peptide delivery and methods of use

Abstract

Disclosed herein are patches, methods, devices, and systems for delivering non-aggregating peptides, such as alcadein and fragments thereof, to a subject. In some aspects, the patch includes a backing, a matrix comprising a non-aggregating peptide disposed within the matrix, and a release liner. In other aspects, methods include opening at least one channel in the skin of a subject, applying the patch described herein, thereby treating a brain-related disease or condition, such as Alzheimer's disease.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 350,982, filed on June 10, 2022, which is incorporated by reference in its entirety.

[0003] Reference sequence list

[0004] This application is submitted in electronic format together with a sequence listing. The sequence listing is provided as a file titled PPORT.008WO_ST_26.xml, which was created on June 6, 2023 and is approximately 7kb in size. The information in the electronic format of the sequence listing is incorporated herein by reference in its entirety. Background of the Invention

[0006] field

[0007] The present application relates to compositions, devices and methods for transdermal drug delivery, and in particular to peptide compositions and methods of administering the peptides to a subject via a transdermal microporation device.

[0008] describe

[0009] Alzheimer's disease (AD) is the most common senile dementia disease. The increase in AD patients is a serious problem in the world. Familial gene mutations that increase the risk of developing AD have been found, but most cases are considered to be due to cognitive decline caused by age-related changes in the brain. Therefore, although aging is the biggest risk factor for developing AD, a common characteristic change in AD patients is the accumulation of β-amyloid protein (Aβ) in the brain. Aβ increases with age, forming aggregates of Aβ oligomers in the brain and producing cytotoxicity. Since it has been reported that Aβ oligomers cause cognitive decline, Aβ is considered to be a strong cause of AD onset.

[0010] The neurospecific membrane protein Alcadeinβ is metabolized to produce p3-Alcβ by a pathway similar to Aβ. Unlike Aβ, which accumulates exponentially in the brain as it ages, p3-Alcβ does not accumulate in the brain, decreases from cerebrospinal fluid (CSF) as it ages, and p3-Alcβ in the CSF of AD patients is further significantly reduced compared to age-matched non-demented subjects, so it is considered to be involved in the onset of AD. In addition, unlike Aβ, p3-Alcβ does not have aggregates, and p3-Alcβ alleviates neurotoxicity caused by Aβo and improves cognitive function by peripheral administration in acute cognitive function model mice induced by Aβ oligomers. It has been clarified that p3-Alcβ shows a reduction and improvement effect. Therefore, the increase of toxic Aβ oligomers and the reduction of p3-Alcβ with an inhibitory effect are considered to be involved in the onset and progression of AD. Since the action of p3-Alcβ is accomplished through its partial peptide p3-Alcβ9-19, the onset and progression of AD associated with the reduction of endogenous molecule p3-Alcβ can be prevented by allowing p3-Alcβ9-19 to reach the brain.

[0011] Various peptide compositions and treatments for AD have been proposed. However, peptide drugs are generally unstable in vivo and difficult to deliver to the brain. Therefore, there is still a long-standing need for improved peptides, formulations, compositions, devices and methods for delivering peptides to cross the blood-brain barrier. Summary of the invention

[0012] Some aspects of the present disclosure relate to a patch for delivering a non-aggregating peptide to a subject. In some embodiments, the patch includes a backing, a matrix, and a release liner, the matrix comprising a non-aggregating peptide disposed within the matrix, wherein the release liner is configured to be removed before application to the skin of the subject. 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-Alcβ is p3-Alcβ9-19 or a derivative thereof. In some embodiments, the amount of the non-aggregating peptide ranges from about 0.01 mg / cm 2 Up to 200mg / cm 2. In some embodiments, the matrix further comprises at least one sugar. In some embodiments, at least one sugar is selected from non-reducing sugars, reducing sugars or a combination thereof. In some embodiments, non-reducing sugars are selected from sucrose, trehalose, mannitol, sorbitol or a combination thereof. In some embodiments, reducing sugars are selected from lactose, maltose or a combination thereof. In some embodiments, the ratio of at least one sugar to non-aggregating peptides is greater than 0.02. In some embodiments, the ratio of at least one sugar to non-aggregating peptides is about 0.02 to about 0.4. In some embodiments, the matrix further comprises an organic acid, an organic salt or a combination thereof. In some embodiments, the organic acid is a pharmacokinetic (PK) modifier. In some embodiments, the PK modifier is citric acid and its salt form. 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 methyl paraben, propyl paraben, benzalkonium chloride and sodium benzoate. In some embodiments, the matrix comprises at least one fiber. In some embodiments, at least one fiber is a nonwoven fiber. In some embodiments, at least one fiber has a thickness of less than 300 μm. In some embodiments, at least one fiber has a thickness of less than 100 g / m 2 In some embodiments, the matrix has a weight of less than 10 mg / cm 2 In some embodiments, the matrix comprises a laminate of films 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.

[0013] Some aspects relate to methods for treating a disease or condition associated with the brain of a subject. In some embodiments, the method includes opening at least one channel in the skin of the subject and applying a patch as described herein to the skin of the subject, 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 skin of the subject includes applying a percutaneous microporation device to the skin of the subject. In some embodiments, the percutaneous microporation device utilizes thermal tissue cauterization by using a filament array having a plurality of filaments, the filaments being disposed in the skin of the subject, wherein each filament is capable of conductively transferring heat energy via direct contact with a tissue membrane to form a plurality of micropores in the microporous region of the tissue membrane. In some embodiments, the percutaneous microporation device produces about 25 to about 500 micropathways / cm 2In some embodiments, the percutaneous microporation device has a perforation energy of about 2 to about 6 mJ / filament. In some embodiments, at least one channel is opened in the skin of the subject with a diameter of about 0.25 cm 2 To about 4cm 2 In some embodiments, the percutaneous microporation device is a microneedle, laser, or radiofrequency device capable of creating one or more micropores in the skin of a subject. In some embodiments, application of the patch increases neuronal activity in a subject. In some embodiments, application of the patch increases mitochondrial activity in the brain of a subject. In some embodiments, the patch provides a maximum blood concentration of non-aggregated peptides for at least 0.5 hours after administration. In some embodiments, the non-aggregated peptides are maintained for at least 6 hours after application of the patch. In some embodiments, after the patch is applied to a subject, the non-aggregated peptides from the patch are maintained in the blood of the subject for at least 6 hours. In some embodiments, the transfer rate of non-aggregated peptides from the blood of a subject to the cerebrospinal fluid of the subject is at least 2%.

[0014] Some aspects relate to a transdermal delivery system for delivering non-aggregated peptides to a subject. In some embodiments, the system includes a patch and a transdermal microporation device as described herein. In some embodiments, the transdermal microporation device includes an applicator electrically connected to an array of conductive filaments and a power supply circuit configured to provide current to the transdermal microporation device, wherein the transdermal microporation device is configured to generate thermal energy based on the current flowing through the array of conductive filaments, and provide thermal energy to a biofilm placed adjacent to the transdermal microporation device. In some embodiments, the applicator provides predetermined electrical energy to the array of conductive filaments, thereby generating one or more micropores. In some embodiments, the patch is applied to one or more micropores. In some embodiments, the transdermal microporation device generates about 25 to about 500 microchannels / cm 2 In some embodiments, the percutaneous microporation device has a perforation energy of about 2 to about 6 mJ / filament. In some embodiments, at least one channel is opened in the skin of the subject with a diameter of about 0.25 cm 2 To about 4cm 2 In some embodiments, the one or more micropores account for about 0.5% to about 12.5% ​​of the total perforated area. In some embodiments, the transdermal microporation device is a microneedle, laser, or radiofrequency device capable of creating one or more micropores in the skin of a subject. In some embodiments, the transdermal microporation device creates at least 50 holes in the skin of a subject. In some embodiments, the patch further includes a drug pellet.

[0015] BRIEF DESCRIPTION OF THE DRAWINGS

[0016] 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 present disclosure. In all drawings, the same numerals represent the same elements.

[0017] Figure 1 The amino acid sequences of Alcβ and p3-Alcβ are shown.

[0018] Figure 2A An example of a patch is shown. Figure 2B The use of percutaneous microporation to apply a patch containing a peptide drug is shown. After microporation of the skin, microchannels are created in the stratum corneum and epidermis. A dry patch containing the peptide drug in the matrix is ​​applied to the microporous area. The dissolved drug migrates into the body via the microchannels.

[0019] Figure 3A Shown is a line graph depicting the PK profile of p3-Alcβ9-19 following intravenous administration in rats; Figure 3B A line graph depicting the PK profile of p3-Alcβ9-19 after subcutaneous administration is shown; Figure 3C A logarithmic scale graph depicting the PK profile of p3-Alc[beta]9-19 is shown.

[0020] Figure 4A Shown is a line graph depicting the dose response of p3-Alcβ9-19 following intravenous administration in rats; Figure 4B Shown is a line graph depicting the dose response of p3-Alcβ9-19 following subcutaneous administration in rats.

[0021] Figure 5A A line graph depicting IR formulations is shown; Figure 5B Shown is a line graph depicting the dose response from percutaneous microporation.

[0022] Fig. 6A Shown are graphs on a logarithmic scale depicting changes in blood and cerebrospinal fluid concentrations of p3-Alcβ9-19 following administration of an IR formulation containing p3-Alcβ9-19 in mice using percutaneous microporation (400 density, 3 mJ / filament). Figure 6B Shown are graphs on a logarithmic scale depicting changes in blood and cerebrospinal fluid concentrations of p3-Alcβ9-19 following administration of a SR formulation comprising p3-Alcβ9-19 in mice using percutaneous microporation (100 density, 3 mJ / filament).

[0023] Figure 7 Shown are two images depicting changes in mitochondrial activity in the brain following percutaneous microporation of p3-Alc[beta]9-19 in monkeys.

[0024] Figure 8Shown are two bar graphs depicting the intensity of mitochondrial activity in various parts of the brain following percutaneous microporation of p3-Alc[beta]9-19 (data analyzed from Figure 6).

[0025] Fig. 9 Shown are bar graphs depicting the percent increase in mitochondrial activity in various parts of the brain following percutaneous microporation of p3-Alcβ9-19 (corresponding to 0.5 mg / kg and 1.0 mg / kg doses of p3-Alcβ9-19).

[0026] Fig. 10A A line graph depicting the PK profile using a non-reducing sugar formulation and the concentration of p3-Alcβ over time is shown; Fig. 10B A logarithmic scale graph depicting the change in p3-Alcβ concentration over time is shown.

[0027] Fig.11 Shown is a line graph depicting the enhancement of β-Alcβ uptake by non-reducing and reducing sugars.

[0028] Fig. 12A A line graph depicting microperforation conditions for controlling PK profiles using reducing sugars with or without PK modifiers is shown; Fig. 12B Shown are logarithmic scale graphs depicting microperforation conditions controlling PK profiles using reducing sugars with or without PK modifiers.

[0029] Fig.13 Shown is a bar graph depicting the effect of microporation conditions on p3-Alcβ9-19 delivery.

[0030] Fig.14 The effect of enhancer content (reducing sugar) in p3-Alc[beta] transdermal microporation is shown.

[0031] Fig.15 A line graph comparing AUC versus lactose content is shown.

[0032] Fig.16 Shown is a line graph depicting AUC versus dose characteristics for the IR formulation.

[0033] Fig.17 Shown is a line graph depicting AUC versus dose characteristics for SR formulations.

[0034] Fig.18 Shown is a line graph depicting the PK profile of an optimized transdermal microporation formulation for immediate delivery.

[0035] Fig.19A A line graph depicting the dose dependence (AUC) of the p3-Alcβ9-19 patch in rats is shown; Fig.19BShown is a line graph depicting the dose dependence (Cmax) of the p3-Alcβ9-19 patch in rats. DETAILED DESCRIPTION

[0036] The present disclosure may be more readily understood by reference to the detailed description, examples, drawings, and claims below, and their previous and subsequent descriptions. However, before disclosing and describing the apparatus, system, and / or method of the present application, it should be understood that, unless otherwise stated, the present disclosure is not limited to the specific apparatus, system, and / or method disclosed. It should also be understood that the terms used herein are only used for the purpose of describing specific aspects and are not necessarily intended to be limiting.

[0037] This specification is provided as an implementation teaching of the present disclosure. For this reason, those skilled in the relevant art will recognize and understand that many changes can be made to the various aspects of the disclosure described herein, while still obtaining beneficial results. It will also be apparent that some desired benefits can be obtained by selecting some features described herein without utilizing other features. Therefore, those skilled in the art will recognize that many modifications and adaptations to this specification are possible, and even desirable in some cases, and these modifications and adaptations are a part of the present disclosure. Therefore, this specification is provided as an explanation of some principles of the present disclosure, rather than a limitation thereof.

[0038] definition

[0039] 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. Unless otherwise indicated, all patents, applications, published applications, and other publications cited herein are incorporated herein by reference in their entirety. If multiple definitions exist for a term herein, those in this section shall prevail unless otherwise indicated.

[0040] As used throughout, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a filament" may include two or more such filaments unless the context dictates otherwise.

[0041] Ranges may be expressed herein as from "about" one particular value, and / or to "about" another particular value. When such ranges are expressed, another aspect includes from a particular value and / or to another particular value. Similarly, when values ​​are expressed as approximations by using the antecedent "about," it will be understood that the particular values ​​form another aspect. It will be further understood that the endpoints of each range are important both relative to the other endpoint and independently of the other endpoint.

[0042] 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 instances where said event or circumstance occurs and instances where it does not.

[0043] As used herein, "stratum corneum" refers to the outermost layer of the skin, consisting of about 15 to about 20 layers of cells in various stages of dryness. The stratum corneum provides a barrier to prevent water loss from the interior of the body to the external environment and to prevent attacks from the external environment to the interior of the body.

[0044] As used herein, "tissue" refers to an aggregate of cells of a particular type together with their intercellular substances that form a structural material. In the case of drug delivery to or through such tissue, a transdermal delivery method (e.g., a perforation device and / or a patch) can access at least one surface of the tissue. The tissue is the skin for various perforation delivery methods described herein. Other tissues suitable for the present disclosure include mucosal tissues and soft organs.

[0045] As used herein, the term "interstitial fluid" is the clear fluid that occupies the spaces between cells in the body.

[0046] As used herein, the term "biological fluid" is defined as a fluid derived from a biological organism, including serum or whole blood, and interstitial fluid.

[0047] As used herein, "tissue membrane" can be any one or more epidermal layers of an object. For example, in one aspect, a tissue membrane is a skin layer including the outermost layer (i.e., stratum corneum) of the skin. In an optional aspect, the skin layer can include one or more backing layers of the epidermis, generally identified as the stratum granulosum, the stratum malpighii, and the stratum germinativum. It will be understood by those of ordinary skill in the art that the transport or absorption of the permeate through the backing layer of the epidermis is substantially free of or free of resistance. Therefore, in one aspect, at least one channel formed in the skin layer of an object is a channel in the stratum corneum of the object. In addition, as used herein, "stratum corneum" refers to the outermost layer of the skin, generally comprising about 15 to about 20 layers of cells in various stages of drying. The stratum corneum provides a barrier to prevent water from being lost from the inside of the body to the external environment and to prevent attacks from the external environment to the inside of the body. In addition, as used herein, "tissue membrane" can refer to an aggregate of cells of a specific type forming a structural material together with their intercellular substances. In various embodiments, one or more perforating devices and / or permeation compositions described herein can approach at least one surface of a tissue membrane. As described above, the tissue membrane used for various perforation delivery methods is the skin. Other tissues suitable for use with such devices and compositions include mucosal tissues and soft organs.

[0048] As used herein, the term "subcutaneous fluid" may include, but is not limited to, water, plasma, blood, one or more proteins, interstitial fluid, and any combination thereof. In one aspect, the subcutaneous fluid according to the present specification is a moisture source comprising water.

[0049] As used herein, "perforation", "microperforation" or any such similar terms means the formation of small holes or cracks (hereinafter also referred to as "micropores") in or through a tissue or biological membrane, such as the skin or mucous membrane, or the outer layer of an organism, to reduce the barrier properties of the biological membrane for the passage of at least one permeate from one side of the biological membrane to the other side for a selected purpose. Preferably, the diameter of the hole or "micropore" so formed is about 1-1000 microns and extends into the biological membrane sufficient to disrupt the barrier properties of the stratum corneum without adversely affecting the underlying tissue. It should be understood that for simplicity, the term "micropore" is used in the singular, but the microporous device described herein can form a plurality of artificial openings. For a selected purpose, or for certain medical or surgical procedures, perforation can reduce the barrier properties of the biological membrane to enter the body. For the purposes of this application, "perforation" and "microperforation" are used interchangeably and mean the same thing.

[0050] A "microporator" or "porator" is a component for a microporation device capable of microporation. Examples of microporators or perforators include, but are not limited to, a filament capable of conductively transferring thermal energy via direct contact with a biological membrane to cause a portion of the membrane to burn deep enough to form micropores, an optically heated local dye / absorbent layer, an electromechanical actuator, a microlancet, an array of microneedles or lancets, an acoustic energy cauterizer, a laser cauterization system, a high-pressure fluid jet puncturer, and the like. As used herein, "microporator" and "porator" are used interchangeably.

[0051] As used herein, "permeation" means the controlled removal of cells caused by the heat and kinetic energy released when the pyrotechnic element explodes, which causes the cells of the biofilm and possibly some neighboring cells to be "blown away" from the scene. As used herein, "fusible" and "fuse" refer to an element that can remove itself from an electrical circuit when a sufficient amount of energy or heat is applied to it. That is, when a resistive electrically activated perforating element is designed as a fusible element, this means that upon activation, during or after the formation of micropores in the biofilm, the element breaks, thereby preventing current from flowing through the element.

[0052] As used herein, "penetration enhancement" or "permeation enhancement" means an increase in the permeability of a biological membrane and / or tissue to a drug, biologically active composition, or other chemical molecule, compound, particle, or substance (also referred to as a "permeant") so as to increase the rate at which the drug, biologically active composition, or other chemical molecule, compound, or particle penetrates the biological membrane and / or tissue.

[0053] As used herein, "enhancer", "chemical enhancer", "penetration enhancer", "permeation enhancer", etc. include all enhancers that increase the flux of permeates, analytes or other molecules across biological membranes and are limited only by functionality. In other words, it is intended to include all cell envelope disordering compounds and solvents as well as any other chemical enhancers. In addition, all active force enhancement techniques are included, such as the application of acoustic energy, mechanical suction, pressure or local deformation of tissue, iontophoresis or electroporation. One or more enhancer technologies can be combined sequentially or simultaneously. For example, a chemical enhancer can be applied first to penetrate the capillary wall, and then iontophoresis or an acoustic energy field can be applied to actively drive the permeate into those tissues surrounding and including the capillary bed.

[0054] As used herein, "transdermal" or "percutaneous" means the passage of a permeant into and through a biological membrane to achieve therapeutically effective blood concentrations or local tissue levels of the permeant, or the passage of molecules or fluids ("analytes") present in the body out of a biological membrane so that the analyte molecules can be collected outside the body.

[0055] 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 by methods taught in this specification that induces a desired biological or pharmacological effect, which may include, but is not limited to (1) having a prophylactic effect on an organism and preventing undesirable biological effects, such as infection, (2) alleviating a condition caused by a disease, such as reducing 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 may be systemic. Such substances include a wide variety of compounds that are typically delivered into the body, including through body surfaces and membranes, including the skin. In general, by way of example and without limitation, such substances may include any biologically active agent, such as a drug, chemical material, or biological material, that induces a desired biological or pharmacological effect. To this end, in one aspect, the permeant may be a small molecule agent. In another aspect, the permeant may be a macromolecular agent.

[0056] In various embodiments, systems, devices, and methods that can be used and / or adapted for use with the compositions and methods described herein are described in one or more of U.S. Pat. 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 entireties and specifically for the purpose of describing such systems and methods. In various embodiments, the compositions and methods described herein may be used in combination with the compositions and methods described herein. Commercially available systems and devices can be used or adapted to deliver the compositions described herein.

[0057] As used herein, an "effective" amount of a pharmacologically active agent means a sufficient amount of the compound to provide the desired local or systemic effect and performance at a reasonable benefit / risk ratio in any medical treatment. An "effective" amount of a permeate or chemical enhancer as used herein means an amount selected to provide the desired increase in biomembrane permeability, the desired depth of penetration, the rate of administration, and the amount of drug delivery.

[0058] As used herein, "animal" or "organism" refers to humans and other living organisms, including plants, to which the present disclosure may be applied.

[0059] As used herein, "analyte" means any chemical or biological material or compound suitable for passing through a biological membrane by the techniques taught in the present disclosure or by techniques previously known in the art, and an individual may wish to know the concentration or activity of the chemical or biological material or compound in vivo. Glucose is a specific example of an analyte because it is a sugar suitable for passing through the skin, and an individual (e.g., an individual with diabetes) may wish to know their blood sugar level. Other examples of analytes include, but are not limited to, compounds such as sodium, potassium, bilirubin, urea, ammonia, calcium, lead, iron, lithium, salicylates, etc.

[0060] 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 into or through the skin of an organism.

[0061] As used herein, "non-invasive" means not requiring the passage of a needle, catheter, or other invasive medical device into a part of the body.

[0062] As used herein, "minimally invasive" refers to the use of mechanical, hydraulic, or electrical devices to penetrate the stratum corneum to create small holes or micropores without causing substantial damage to the underlying tissue.

[0063] As used herein, "pharmaceutically acceptable carrier" refers to a carrier in which a substance for delivery (e.g., a pharmaceutically acceptable drug) can be provided. 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 (e.g., polyesters, polyanhydrides, polyamino acids, liposomes, and mixtures thereof).

[0064] As used herein, "reservoir" refers to a designated area or chamber in a device that is designed to hold a permeate for delivery to an organism through an artificial opening in a biofilm, or can be designed to receive a biological fluid sample extracted from an organism through an artificial opening in a biofilm. The reservoir can also hold excipient compounds that enhance the effect of a biologically active permeate contained alone. In addition, the reservoir can hold a reactive enzyme or reagent that is designed to allow measurement or detection of a selected analyte in the extracted biological fluid, or can be treated with the reactive enzyme or reagent. The reservoir can include an open volume space, a gel, a planar space (which has been coated or treated with a selected compound for subsequent release or reaction), or a matrix or a permeable solid structure (e.g., a pellet, a tablet, a powder, a dry solid or a porous polymer).

[0065] As used herein, "p3-Alcβ" will be produced as a metabolite of the membrane protein Alcadeinβ (Alcβ). Alcβ is metabolized by a pathway similar to Aβ to produce p3-Alcβ. Unlike Aβ accumulated in the brain, p3-Alcβ is more significantly reduced in Alzheimer's patients. The effect of p3-Alcβ is achieved through its partial peptides, such as p3-Alcβ1-40, p3-Alcβ1-37, p3-Alcβ9-19, p3-Alcβ1-19, p3-Alcβ11-19 or its derivatives.

[0066] Patches

[0067] In various aspects, the systems, devices and methods of the present disclosure can be used to deliver peptides transdermally through the skin. In some aspects, the patch can include a top layer comprising an adhesive, an intermediate layer comprising a matrix, and a bottom layer. In some embodiments, the bottom layer includes a release liner. In some embodiments, the intermediate layer also includes a PK modifier. In some embodiments, the intermediate layer also includes a reinforcing agent. In some embodiments, the patch includes a tissue interface layer. In some embodiments, the patch also includes a backing. In some embodiments, the release liner is configured to be removed before being applied to the skin of the object. In some embodiments, the patch is configured as a film. In some embodiments, the patch is configured as a pellet.

[0068] Examples of suitable tissue interface layers are described in US Pat. No. 7,392,080, which is hereby incorporated by reference in its entirety and particularly for the purpose of describing transdermal drug delivery patch systems.

[0069] In some embodiments, the top layer includes a backing. In some embodiments, the backing is a film, form, woven or nonwoven material. In some embodiments, the film includes polyethylene (PE), polyethylene terephthalate (PET), polyurethane (PU), polyvinyl chloride (PVC), polychlorotrifluoroethylene (PCTFE), cycloolefin copolymer (COC) or polymer (COP). In some embodiments, the backing includes an adhesive. In some embodiments, the adhesive is an acrylic acid, silicone or synthetic rubber, such as polyisobutylene (PIB) and styrene-isoprene-styrene block copolymer (SIS). In some embodiments, the color of the backing is transparent, translucent, brown, white or beige. In some embodiments, the backing is formed by thermoforming to make a cavity. In some embodiments, the backing is covered with adhesive tape.

[0070] In some embodiments, the bottom 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 rubber coating. In some embodiments, the release liner is heat sealed to the backing film.

[0071] In some embodiments, the PK modifier is a drug delivery modifier. In some embodiments, the PK modifier is a pH control agent. In some embodiments, the PK modifier is an organic acid, a salt form of an organic acid, or a combination of the above. 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, an amino acid, or a combination of the above. In some embodiments, the PK modifier is a non-organic acid. In some embodiments, the PK modifier is a salt form of a non-organic acid. In some embodiments, the non-organic acid is hydrochloric acid, phosphoric acid, boric acid, acetic acid, or a combination of the above. In some embodiments, the non-organic 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 of the above.

[0072] In some aspects, the enhancer is a carbohydrate. In some embodiments, the carbohydrate comprises sugar or is a sugar. In some embodiments, the enhancer is a non-reducing sugar. In some embodiments, the enhancer is a reducing sugar. In some embodiments, the carbohydrate 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.

[0073] In some embodiments, the weight ratio of the enhancer to the non-aggregating peptide is greater than 0.005, greater than 0.02, greater than 0.05, greater than 0.1, greater than 0.2, or a range including and / or spanning the above values. In some embodiments, the weight ratio of the sugar to the non-aggregating peptide is greater than 0.02. In some embodiments, the weight ratio of the sugar to the non-aggregating peptide is about 0.02 to about 10. In some embodiments, the weight ratio of the sugar to the non-aggregating peptide is about 0.02 to about 0.4.

[0074] In some aspects, the middle layer of the patch includes a reservoir. In some embodiments, the reservoir includes about 1.0% to about 99.5% non-aggregated peptide by weight. In some embodiments, the non-aggregated peptide accounts for about 50% to about 98% by weight of the middle layer, including, for example, 50%, 55%, 70%, 80%, 90%, 95%, 98% by weight of the middle layer, including any weight percentage range derived from these values.

[0075] In some embodiments, the intermediate layer comprises from about 0.5% to about 99% reinforcing agent by weight. In some embodiments, the reinforcing agent comprises from about 2% to about 50% by weight of the intermediate layer, including, for example, 2%, 5%, 10%, 15%, 30%, 40%, 50% by weight of the intermediate layer, and including any weight percentage range derived from these values.

[0076] In some embodiments, the intermediate layer comprises about 5% to about 60% by weight of the PK modifier. In some embodiments, the PK modifier comprises about 10% to about 40% by weight of the intermediate layer, including, for example, additional amounts of 7.5%, 15%, 25%, 40% by weight of the intermediate layer, and including any weight percentage range derived from these values.

[0077] In some embodiments, the intermediate layer comprises at least 0.5 mg / cm2 of the surface area of ​​the intermediate layer facing the bottom layer. 2 , at least 1mg / cm2 , at least 2mg / cm 2 , at least 4 mg / cm 2 , at least 8mg / cm 2 Or a PK modifier including and / or spanning the above values. In some embodiments, the middle layer comprises at least 2 mg / cm 2 In some embodiments, the middle layer comprises at least 4 mg / cm 2 of disodium citrate.

[0078] In some aspects, the middle layer includes a matrix support. In some embodiments, the matrix support includes 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 laminate of film. In some embodiments, the film is polyethylene terephthalate. In some embodiments, the matrix support is a laminate of fiber. In some embodiments, the matrix is ​​a laminate of film and fiber.

[0079] 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 a range including and / or spanning the above values.

[0080] In some embodiments, the fiber has an area weight of less than 100 g / m 2 Less than 90g / m 2 Less than 80g / m 2 Less than 70g / m 2 Less than 60g / m 2 Less than 50g / m 2 Less than 40g / m 2 Less than 30g / m 2 Less than 20g / m 2 Less than 10g / m 2 , or a range including and / or spanning the above values.

[0081] In some embodiments, the matrix support has a surface area of ​​about 0.1 mg / cm 2 To about 10mg / cm 2 The water holding capacity (WHC) of the matrix support refers to the water holding capacity of the matrix support per 1 cm 2 The amount of water that the matrix support can retain on the transdermal surface. Specifically, a 1 cm 2 substrate and immersed it in a solution (containing 0.1% surfactant ( 80) phosphate buffered saline) for a sufficient period of time. The matrix support is then slowly pulled out of the solution for about 5 seconds, and the pre-measured weight of the sample before immersion is subtracted from the weight of the sample holding the liquid, and the mass per unit area (1 cm 2 ) of the transdermal surface of the matrix. In some embodiments, the matrix support has a water holding capacity of about 10 mg / cm 2 In some embodiments, the water holding capacity of the matrix support is about 1 mg / cm 2 About 8mg / cm 2 In some embodiments, the matrix water holding capacity is about 2 mg / cm 2 To about 5mg / cm 2 .

[0082] The water holding capacity of the matrix support can be controlled by adjusting the thickness and weight of the matrix support. Preferably, the matrix support has a thickness of 100 μm or less. In some embodiments, the thickness range of the matrix support is from about 10 μm to about 100 μm. In some embodiments, the matrix support thickness is from about 20 μm to about 90 μm. In some embodiments, the matrix support thickness is from about 30 μm to about 80 μm. In some embodiments, the matrix support thickness is from about 40 μm to about 60 μm.

[0083] In some embodiments, the matrix support area weight is about 10 g / m 2 About 100g / m 2 In some embodiments, the matrix support area weight is about 15 g / m 2 About 80g / m 2 In some embodiments, the matrix support area weight is about 20 g / m 2 About 60g / m 2 In some embodiments, the matrix support area weight is about 25 g / m 2 About 40g / m 2 .

[0084] In some embodiments, the matrix support area weight is about 0.1 mg / cm 2 To about 30mg / cm 2 In some embodiments, the matrix support area weight is about 0.5 mg / cm 2 To about 30mg / cm 2 In some embodiments, the matrix support area weight is about 0.5 mg / cm 2 To about 20mg / cm 2 In some embodiments, the matrix support area weight is about 0.5 mg / cm 2To about 10mg / cm 2 .

[0085] In some embodiments, the size of the matrix is ​​about 0.125 cm 2 To about 4cm 2 In some embodiments, the size of the matrix is ​​about 0.25 cm 2 About 3cm 2 In some embodiments, the size of the matrix is ​​about 0.5 cm 2 About 2cm 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 .

[0086] In some embodiments, the total amount of non-aggregated peptide and enhancer per unit area of ​​matrix (surface area) is 0.01 mg / cm 2 Up to 200mg / cm 2 In some embodiments, the total amount of non-aggregated peptide and enhancer per unit area of ​​matrix is ​​0.1 mg / cm 2 Up to 100mg / cm 2 In some embodiments, the total amount of non-aggregated peptide and enhancer per unit area of ​​matrix is ​​5 mg / cm 2 Up to 75mg / cm 2 In some embodiments, the total amount of non-aggregated peptide and enhancer per unit area of ​​matrix is ​​10 mg / cm 2 Up to 50mg / cm 2 .

[0087] 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.

[0088] In some aspects, the substrate comprises about 0.01 mg / cm based on the surface area of ​​the substrate facing the bottom layer. 2 To about 200mg / cm 2 In some embodiments, the matrix comprises about 0.1 mg / cm 2 To about 100mg / cm 2 In some embodiments, the matrix comprises about 1 mg / cm 2 To about 50mg / cm 2In some embodiments, the matrix comprises about 5 mg / cm 2 To about 30mg / cm 2 In some embodiments, the matrix comprises about 5 mg / cm 2 To about 25mg / cm 2 In some embodiments, the matrix comprises about 0.5 mg / cm 2 In some embodiments, the matrix comprises about 1 mg / cm 2 In some embodiments, the matrix comprises about 2 mg / cm 2 In some embodiments, the matrix comprises about 3 mg / cm 2 In some embodiments, the matrix comprises about 5 mg / cm 2 In some embodiments, the matrix comprises about 10 mg / cm 2 In some embodiments, the matrix comprises about 20 mg / cm 2 In some embodiments, the matrix comprises about 25 mg / cm 2 In some embodiments, the matrix comprises about 50 mg / cm 2 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 derivatives thereof. In some embodiments, p3-AlcB is p3-Alcβ9-19 or derivatives thereof.

[0089] In some aspects, the substrate comprises about 0.05 mg / cm based on the area of ​​the substrate facing the bottom layer. 2 To about 100mg / cm 2 In some embodiments, the matrix comprises about 0.1 mg / cm 2 To about 50mg / cm 2 In some embodiments, the matrix comprises about 0.2 mg / cm 2 To about 5.0mg / cm 2 In some embodiments, the matrix comprises about 0.5 mg / cm 2 To about 10mg / cm 2 In some embodiments, the matrix comprises about 0.5 mg / cm 2 to about 4.0 mg / cm 2In some embodiments, the matrix comprises about 0.5 mg / cm 2 To about 2.0mg / cm 2 In some embodiments, the matrix comprises about 0.5 mg / cm 2 In some embodiments, the matrix comprises about 1 mg / cm 2 In some embodiments, the matrix comprises about 2 mg / cm 2 In some embodiments, the enhancer is at least selected from sucrose, lactose and maltose.

[0090] In some aspects, 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.

[0091] In some embodiments, the patch is configured as a dry patch preparation. In some embodiments, the dry patch includes a backing, a matrix containing non-aggregated peptides in a dry state, and a release liner. In some embodiments, the matrix also includes a PK modifier in a dry state. In some embodiments, the matrix also includes an enhancer in a dry state. In some embodiments, the matrix also includes an antimicrobial in a dry state. In some embodiments, the dry patch is a heat-dried film made by a distribution or casting process. In some embodiments, the dry patch is a tablet or pellet made by a compression process. In some embodiments, the dry patch preparation includes p3-Alcβ9-19, sucrose, lactose, disodium citric acid sesquihydrate, methyl paraben, propyl paraben, sodium benzoate, and benzalkonium chloride. In some embodiments, the dry state includes less than 10% of the water in the preparation. In some embodiments, the dry state includes less than 5% of the water in the preparation. In some embodiments, the dry state includes less than 2% of the water in the preparation. In some embodiments, the dry state includes less than 1% of the water in the preparation.

[0092] In some embodiments, the patch is configured as a freeze-dried patch preparation. In some embodiments, the freeze-dried patch includes a backing, a matrix containing p3-Alcβ9-19 in a freeze-dried state, and a release liner. In some embodiments, the freeze-dried patch consists of a cavity with a spacer. In some embodiments, the freeze-dried patch consists of a blister package. In some embodiments, the matrix also includes a drug delivery modifier in a freeze-dried state. In some embodiments, the matrix includes an enhancer in a freeze-dried state. In some embodiments, the matrix also includes an antimicrobial in a freeze-dried state. In some embodiments, the freeze-dried patch is freeze-dried by methods known to those skilled in the art. In some embodiments, the freeze-dried patch is a tablet or pellet manufactured by a compression process. In some embodiments, the freeze-dried patch preparation includes p3-Alcβ9-19, sucrose, lactose, disodium citrate sesquihydrate, methyl paraben, propyl paraben, sodium benzoate, and benzalkonium chloride. In some embodiments, the freeze-dried patch includes less than 10% water in the formulation. In some embodiments, the lyophilized patch includes less than 5% water in the formulation. In some embodiments, the lyophilized patch includes less than 2% water in the formulation. In some embodiments, the lyophilized patch includes less than 1% water in the formulation. In some embodiments, the lyophilized patch includes less than 0.5% water in the formulation. In some embodiments, the lyophilized patch includes less than 0.1% water in the formulation.

[0093] 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 various sheet-like 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, a substantially flat thin solid tablet is slightly curved or bent to a degree that is convenient to pick up (e.g., compared to a flat thin solid tablet that is more difficult to pick up from a flat surface). In various embodiments, the thin solid tablet as described herein has the following area density: greater than 30 mg / cm 2 , greater than 40mg / cm 2 , greater than 50mg / cm 2 , greater than 60mg / cm 2 , greater than 70mg / cm 2 , greater than 80mg / cm 2 , greater than 90mg / cm 2 Or greater than 100mg / cm 2 ; less than 400mg / cm 2 Less than 350mg / cm 2 Less than 300mg / cm 2 Less than 250mg / cm 2Or less than 200mg / cm 2 ; or within any range having endpoints defined by any two of the above values. For example, in various embodiments, the thin solid tablet has greater than 30 mg / cm 2 And less than 400mg / cm 2 ; greater than 40mg / cm 2 And less than 400mg / cm 2 ; or greater than 30 mg / cm 2 And less than 400mg / cm 2 area density.

[0094] In various embodiments, the thin solid tablet as described herein has the following thickness (depending on the area density and area of ​​the face): 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 0.05 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 in any range with endpoints defined by any two of the above values. For example, in various embodiments, the thickness of the thin solid tablet ranges from about 0.01 mm to about 10 mm or from about 0.1 mm to about 5 mm.

[0095] In various embodiments, the thin solid tablet has a face in a manner similar to the front or back of a coin. In various embodiments, the face of the thin solid tablet has an area of ​​about 0.01 cm 2 or larger, about 0.05cm 2 or larger, about 0.1cm 2 or larger, about 0.25cm 2 or larger, about 0.5cm 2 or larger, about 0.75cm 2 or larger or about 1cm 2 or larger; or about 50cm 2 or smaller, about 25cm 2 or smaller, about 15cm 2 or smaller, about 10cm 2 or smaller, about 5cm 2 or smaller or about 2cm 2 or less; or within any range having endpoints defined by any two of the above values. For example, in various embodiments, the area range of the face of the thin solid tablet is about 0.01 cm 2 To about 25cm 2 、About 0.1cm 2 To about 10cm2 or about 0.15cm 2 To about 5cm 2 .

[0096] In some embodiments, the patch is configured as a reservoir patch. In some embodiments, the reservoir patch includes a backing, a spacer forming a cavity, a matrix containing p3-Alcβ9-19 in the cavity, and a release liner. The matrix can be various shapes, such as elliptical, circular, triangular, rectangular, square, pentagonal, hexagonal, irregular, etc. In one embodiment, the depth of the cavity is about 0.5mm to 10mm. In some embodiments, the depth of the cavity is about 0.5mm to 5mm. In some embodiments, the depth of the cavity is 1mm to 3mm.

[0097] In some embodiments, the patch is configured as a freeze-dried dry patch. In some embodiments, the freeze-dried dry patch includes a backing, an intermediate layer and a release liner comprising a matrix containing freeze-dried p3-Alcβ9-19. The matrix can be in various shapes, such as oval, circular, triangular, rectangular, square, pentagonal, hexagonal, irregular, etc. In one embodiment, the matrix is ​​about 0.5mm to 10mm. In some embodiments, the matrix is ​​about 0.5mm to 5mm. In some embodiments, the matrix is ​​1mm to 3mm.

[0098] In some embodiments, the patch is configured as a tablet patch. In some embodiments, the tablet patch includes a backing, an intermediate layer containing a tablet containing p3-Alcβ9-19, and a release liner. The matrix can be various shapes, such as oval, circular, triangular, rectangular, square, pentagonal, hexagonal, irregular, etc. In one embodiment, the matrix is ​​about 0.5mm to 10mm. In some embodiments, the matrix is ​​about 0.5mm to 5mm. In some embodiments, the matrix is ​​1mm to 3mm.

[0099] In some embodiments, the patch is configured as a solid dispersed dry patch. In some embodiments, the solid dispersed dry patch includes a backing, an intermediate layer containing a matrix containing p3-Alcβ9-19 in a dry state, and a release liner. The matrix can be various shapes, such as oval, circular, triangular, rectangular, square, pentagonal, hexagonal, irregular, etc. In one embodiment, the matrix is ​​about 0.5mm to 10mm. In some embodiments, the tablet is about 0.5mm to 5mm. In some embodiments, the matrix is ​​1mm to 3mm.

[0100] Method / Purpose

[0101] Aspects disclosed herein relate to administering an effective amount of a non-aggregated peptide as disclosed elsewhere herein to a subject in need thereof. Some embodiments relate to treating Alzheimer's disease by administering a patch as disclosed herein. In some embodiments, the patch of the present disclosure can deliver the non-aggregated peptide transdermally to a subject through one or more micropores formed by a transdermal delivery system as described herein. In some embodiments, the method may include punching holes in the outermost stratum corneum and epidermis, and then applying a patch thereon, so that the non-aggregated peptide in the patch passes through the epidermis, diffuses into the capillary dermis, and enters the systemic circulation. Non-aggregated peptides absorbed from the skin can avoid first-pass metabolism in the liver and be effectively delivered to the brain. Therefore, embodiments of the patch of the present disclosure can provide a means of drug administration to replace injection, and the patch can be appropriately used for immediate release applications or sustained release of non-aggregated peptides, with higher bioavailability compared to oral, nasal or passive transdermal administration.

[0102] In some embodiments, the patch of the present disclosure can also be used in a method for delivering non-aggregated peptides through a target biological membrane, wherein the method includes a step for forming one or more micropores in the biological membrane and a step for placing the patch in physical contact with the one or more micropores so that at least a portion of the drug can be dissolved in the biological moisture received from the target through the one or more micropores.

[0103] In some embodiments, the method of treating a subject comprises: identifying a subject with a brain disease or condition; opening a plurality of micropores in the subject's skin; and applying the patch to the subject's skin above the micropores for a period of time to effectively cause transdermal delivery of non-aggregated peptides. In some embodiments, the patch comprises a top layer comprising an adhesive, an intermediate layer comprising non-aggregated peptides, and a bottom layer. In some embodiments, the bottom layer comprises a release liner. In some embodiments, a time period is selected to deliver a therapeutically effective amount of non-aggregated peptides through a plurality of micropores.

[0104] In some embodiments, opening a plurality of micropores in the skin of the subject comprises applying a percutaneous microporation device to the skin of the subject. In some embodiments, the percutaneous microporation device comprises a conductive member comprising an array of conductive filaments. In some embodiments, the percutaneous microporation device comprises a conductive member comprising an array of conductive filaments. In some embodiments, the percutaneous microporation opens the micropores by thermal tissue cauterization. In some embodiments, the percutaneous microporation produces micropores through the stratum corneum to the epidermis.

[0105] The patch of the present disclosure can be applied to an object with a disease or condition that requires an immediate effect or a sustained effect. In some embodiments, the patch and transdermal delivery of non-aggregated peptides can have PK characteristics comparable to subcutaneous injection. In some embodiments, the patch and transdermal delivery of non-aggregated peptides can have PK characteristics superior to intravenous and subcutaneous injections. In some embodiments, the patch and transdermal delivery of non-aggregated peptides can have a bioavailability greater than 100%, greater than 200%, greater than 300%, greater than 400%, greater than 500%, greater than 750%, greater than 1000%, greater than 1500%, greater than 2000%, greater than 2500%, greater than 3000%, greater than 3500%, greater than 4000%, greater than 4500%, greater than 5000% or include and / or span the PK characteristics of the range of the above values ​​compared to intravenous (IV) administration. In some embodiments, patch and transdermal delivery of non-aggregating peptides can have PK profiles with bioavailability ranging from about 350% to about 4800% compared to IV.

[0106] In some embodiments, the patch and transdermal delivery of non-aggregated peptides can provide enhanced delivery of non-aggregated peptides through the skin of the subject. In some embodiments, compared with IV or subcutaneous administration, the patch and transdermal delivery of non-aggregated peptides can provide a longer sustained delivery of non-aggregated peptides to the subject. In some embodiments, for non-aggregated peptides, the area under the curve (AUC) of the patch and transdermal delivery of non-aggregated peptides in plasma 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 intravenous injection, or including and / or spanning the range of the above values. In some embodiments, for non-aggregated peptides, the area under the curve (AUC) of the patch and transdermal delivery of non-aggregated peptides in plasma is 5.4 times higher than intravenous injection. For non-aggregated peptides, the area under the curve (AUC) of the patch and transdermal delivery of non-aggregated peptides in plasma is 8.9 times higher than intravenous injection.

[0107] In some embodiments, patch and transdermal delivery of an effective amount of the non-aggregating peptide into the subject's blood is achieved at least 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 a range including and / or spanning the above values.

[0108] In some embodiments, an effective amount of the patch and transdermal delivery of the non-aggregating peptide into the subject's blood 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 including and / or spanning the above values.

[0109] In some embodiments, patch and transdermal delivery of non-aggregated peptides into the cerebrospinal fluid achieves 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 a range including and / or spanning the above values.

[0110] In some embodiments, the transfer of non-aggregated peptide from blood to cerebrospinal fluid (the amount of non-aggregated peptide in cerebrospinal fluid divided by the amount in plasma, expressed as a percentage) 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 including and / or spanning the above values. In some embodiments, the transfer of non-aggregated peptide from blood to cerebrospinal fluid is about 3.5%.

[0111] In some embodiments, a method for mitochondrial activation in the brain is provided. In some embodiments, the method includes opening at least one microchannel in the skin of the subject and applying a patch as disclosed herein to the skin of the subject. In some embodiments, 0.5 hours after administration, 1.0 hours 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 including and / or spanning the above values, BCPP-EF accumulation activation of mitochondria is achieved. In some embodiments, 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 including and / or spanning the above values, mitochondrial activation in the brain is maintained.

[0112] In some embodiments, before obtaining a high level of non-aggregated peptides, the object receives enough non-aggregated peptides from multiple dosages. It is easy and immediately conceivable to have a scheme in which the first patch is applied to the object, and the object receives one or more subsequent patches. Such schemes can be continued so that the object receives the third patch after receiving the second patch. In some embodiments, the object can receive 1,2,3,4,5,6,7,8,9,10 or more patches during treatment. One or more additional patches as described herein can be applied before the first patch dosage or before one or more subsequent patch dosages. In some embodiments, the object receives dosage during the time course of the remainder of his or her life and / or in the time of many years (for example, more than or equal to 1 year, 5 years, 10 years, 15 years, 20 years, 30 years or including and / or spanning the scope of the above values).

[0113] In some cases, a period of time passes between applying one or more patches to a subject. In some embodiments, the time period between applying one or more patches is equal to or at least about: 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 a range including and / or spanning the above values. In some embodiments, one or more additional therapeutic agents are administered to a subject during the time period between applying the composition to the subject.

[0114] Before receiving the first treatment using the patch described herein, it is not always necessary to identify the subject who needs to receive the patch as disclosed herein to improve the health of the subject. For example, before showing any current signs or symptoms of Alzheimer's disease (normal cognition in the preclinical stage) or mild cognitive problems of Alzheimer's disease (mild MCI stage), it can be predetermined that the subject will develop Alzheimer's disease in the future. Alternatively, if the subject is at risk of Alzheimer's disease or (for example, once the patient reaches an age equal to or greater than 50, 60, 70, etc.) is not at risk of Alzheimer's disease, he or she can be treated prophylactically. Therefore, in some embodiments, after the subject receives an early diagnosis, the patch is applied to the subject. In some embodiments, not every subject is a candidate for such administration, and it may be necessary to identify the subject for treatment. It should be understood that the selection of the patient depends on many factors within the skill range of an ordinary skilled physician. Therefore, some embodiments disclosed herein also include identifying the subject as a subject that will benefit from the administration of an effective amount of at least one non-aggregating peptide or a composition comprising the non-aggregating peptide to increase lifespan, increase survival time, increase natural lifespan or improve immunity. According to the age of the subject, the current medical condition, the current medical treatment, the prescribed medical treatment or in some embodiments, the subject diagnosed with Alzheimer's disease, the subject can be identified based on the subject's specific physiological factors. In some embodiments, the treatment of Alzheimer's disease includes preventing, reducing and / or slowing the accumulation of beta-amyloid protein, amyloid plaques and / or tau protein tangles. Beta-amyloid protein is the remaining fragment of a larger protein. When these fragments gather together, they appear to have a toxic effect on neurons and destroy intercellular communication. These aggregates form larger deposits called amyloid plaques, which also include other cell fragments. Tau protein plays a role in the internal support and transport system of neurons to carry nutrients and other essential substances. In Alzheimer's disease, tau protein changes shape and organizes itself into a structure called neurofibrillary tangles. Tangles destroy the transport system and are toxic to cells.

[0115] In some aspects, a method for treating, preventing or improving brain diseases by applying a patch as described herein. In some embodiments, brain diseases are dementia. In some embodiments, dementia is caused by Alzheimer's disease and Parkinson's disease. Dementia is a broad category of brain diseases that causes long-term and usually gradually reduced thinking and memory abilities that are serious enough to affect daily functions. Symptoms associated with dementia include emotional problems, language difficulties and decreased motivation. In some embodiments, brain diseases are neurodegenerative diseases, such as amyotrophic lateral sclerosis (ALS), Huntington's disease, ataxia, spinal muscular atrophy, Lewy body disease. In some embodiments, methods include testing for dementia risk factors of the subject and applying a composition as described herein. In some embodiments, the dementia risk factors of the test subject include performing brain scans and amyloid PET, performing brain biopsies, cognitive tests and test subject blood. According to the age of the subject, the current medical condition, the current medical treatment, the prescribed medical treatment or in some embodiments, the subject diagnosed with dementia can be identified based on the subject's specific physiological factors.

[0116] Some aspects provided herein provide methods for producing ultra-physiologically high levels of non-aggregated peptides in a subject. In some embodiments, the method includes providing a non-aggregated peptide, composition, or patch as disclosed elsewhere herein to a subject. 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 comprises a non-aggregated peptide, an enhancer, a PK modifier, and an antimicrobial. In some embodiments, the enhancer is a reducing sugar. In some embodiments, the PK modifier is an organic acid. In some embodiments, the antimicrobial is a paraben. In some embodiments, the antimicrobial is sodium benzoate. In some embodiments, the bioavailability of the non-aggregated peptide provided by the patch is increased by at least 200%. In some embodiments, the transfer rate of the non-aggregated peptide provided by the method from blood to CSF ​​is about 3.5%.

[0117] Some aspects provide methods for treating or improving diseases, disorders or conditions associated with Alzheimer's disease in a subject; the method comprises administering to the subject a therapeutically effective amount of a non-aggregating peptide, composition or patch as disclosed elsewhere herein. In some embodiments, the patch comprises a non-aggregating peptide, an enhancer, a PK modifier and an antimicrobial agent. In some embodiments, the non-aggregating peptide or composition improves cerebral blood flow in the subject. In some embodiments, the aggregated peptide or composition prevents or inhibits hyperphosphorylation of tau protein. In some embodiments, hyperphosphorylation of tau protein is inhibited by equal to or at least about: 50%, 70%, 80%, 90%, 99% or a range including and / or spanning the above values. In some embodiments, the composition reduces beta-amyloid in the brain of the subject. In some embodiments, the beta-amyloid in the brain of the subject is reduced by equal to or at least about: 50%, 70%, 80%, 90%, 99% or a range including and / or spanning the above values. In some embodiments, the aggregated peptide or composition has neuroprotective and neuroactivating effects.

[0118] In some embodiments, the treatment methods as described herein downregulate the β-secretase activity in the subject. In some embodiments, the β-secretase activity is reduced by equal to or at least about: 50%, 70%, 80%, 90%, 99% or a range including and / or spanning the above values. In some embodiments, the treatment methods as described herein upregulate the α-secretase activity in the subject. In some embodiments, the α-secretase activity increases by equal to or at least about: 150%, 170%, 180%, 190%, 199% or a range including and / or spanning the above values. In some embodiments, the treatment methods as described herein upregulate the enkephalinase activity in the subject. In some embodiments, the enkephalinase activity increases by equal to or at least about: 150%, 170%, 180%, 190%, 199% or a range including and / or spanning the above values.

[0119] In some aspects, using compositions as described herein can increase the life span, survival time, natural life span or healthy life span of the object. In some embodiments, the life expectancy, survival time, natural life span or healthy life span of the object is the median expectation of the object of similar conditions. In other embodiments, the life expectancy, survival time, natural life span or healthy life span of the object is the average expectation of the object of similar conditions. The object of similar conditions 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 of initiation of treatment relative to the subject's expected lifespan, survival time, natural lifespan, or healthy lifespan can be 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%, with respect to any of the above percentages. or a range defined by any of the above 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%, or about any one of the above percentage ranges (e.g., about 10%-70%, about 30%-60%, or about 50%-70%).In some embodiments, the expected increase relative to the subject's expected lifespan, survival time, natural lifespan or healthy lifespan is measured in years and is 1-40 years, 1-19 years, 1-18 years, 1-17 years, 1-16 years, 1-15 years, 1-14 years, 1-13 years, 1-12 years, 1-11 years, 1-10 years, 1-9 years, 1-8 years, 1-7 years, 1-6 years, 1-5 years, 1-4 years, 1-3 years, 1-2 years, 1 year, at least the above number of years (e.g., at least 1-10 years) or about the above number of years (e.g., about 1-2 years or at least about 1-2 years). In some embodiments, the expected increase relative to the subject's expected lifespan, survival time, natural lifespan or healthy lifespan is measured in days to months and is one day to one year, one day to 11 months, one day to 10 months, one day to 9 months, one day to 8 months, one day to 7 months, one day to 6 months, one day to 5 months, one day to 4 months, one day to 3 months, one day to 2 months, one day to 1 month, at least the above range of days to months (e.g., at least one day to 11 months) or about the above range of days to months (e.g., about one day to 6 months or at least about one day to 6 months).

[0120] Some embodiments relate to treating a disease or condition associated with the brain in a subject. In some embodiments, the beta-amyloid 42 / beta-amyloid 40 ratio of a subject is reduced. In some embodiments, the composition improves glucose metabolism in the brain of a subject. In some embodiments, the composition improves cerebral blood flow in a subject. In some embodiments, the composition prevents or substantially prevents hyperphosphorylation of tau protein. In some embodiments, the composition reduces beta-amyloid in the brain of a subject. In some embodiments, the composition reduces the risk of cardiac arrest and stroke in a subject.

[0121] Some embodiments relate to increasing the mitochondrial activity of the brain of a subject. In some embodiments, the mitochondrial activity of the brain of a subject may be increased by at least about 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or a range including and / or spanning the above values. In some embodiments, the mitochondrial activity of the brain of a subject may be restored. In some embodiments, the mitochondrial activity of the brain of a subject 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 a range including and / or spanning the above values.

[0122] In some embodiments, the method for treating cognitive impairment induced by amyloid beta according to the present disclosure may include the step of administering the patch to a mammal (human, or mammal other than human) or a model animal thereof (mammal other than human) suffering from cognitive impairment induced by amyloid beta. In some embodiments, the method for treating Alzheimer's disease according to the present disclosure may include the step of administering the patch as described herein to a mammal (excluding human) or a model animal thereof suffering from Alzheimer's disease.

[0123] Cognitive impairment induced by amyloid beta protein treated with a therapeutic agent according to the present disclosure generally refers to symptoms associated with Alzheimer's disease, but may be symptoms associated with a disease that has not yet been clearly diagnosed as suffering from Alzheimer's disease, or symptoms in subjects or model animals in the preclinical stage (including mild cognitive impairment (MCI)). Model animals for cognitive impairment induced by amyloid beta protein can be prepared using known methods. For example, transgenic mice that exhibit overexpression of amyloid beta protein or mice to which a solution of amyloid beta protein is administered in artificial cerebrospinal fluid can be used as such model animals.

[0124] In some embodiments, as discussed elsewhere herein, the subject is a human. However, the methods are not limited to the treatment of humans, and are equally applicable to the treatment of mammals. In the case of treating non-human mammals, the selection of the patient depends on many factors within the skill of an ordinary skilled veterinarian or research scientist.

[0125] System / Equipment

[0126] The basic configuration of microperforated drug delivery system is known to those skilled in the art, so further detailed description is not required herein.For example, in U.S. Patent No. 8,116,860, a transdermal permeation delivery system is described, which is incorporated herein by reference and particularly for the purpose of describing the various features of such microperforated drug delivery system.As described in the patent, the microperforated drug delivery system of U.S. Patent No. 8,116,860 (referring to using reference number "10" therein) includes the basic features containing filament array (referring to using reference number "70" therein) and one or more transdermal patches (referring to using reference number "100" therein), the filament array is configured to produce one or more channels or micropores in the skin of the patient, and the transdermal patch includes at least one pharmaceutical preparation.Other similar microperforated drug delivery systems including such basic features are known to those skilled in the art.Various microperforated drug delivery systems with such basic features are known to those skilled in the art, and can be used or transformed by those skilled in the art under the guidance of the teachings provided herein.

[0127] In some embodiments, a microporation device can be defined by the total area of ​​channels (e.g., micropores) created in the skin by one or more filaments and the total energy delivered to the one or more filaments to create the channels. In some embodiments, a microporation device creates channels such that the total area of ​​the channels in the skin is approximately 0.25 cm per square centimeter (cm) of skin. 2 Up to 4.0cm 2 In some embodiments, the channels in the skin may preferably comprise approximately 0.5% to 12.5% ​​of the skin area for each square centimeter of skin exposed to the microporation device. In some embodiments, the channels in the skin may preferably comprise approximately 1.25% to 10% of the skin area for each square centimeter of skin exposed to the microporation device.

[0128] In some embodiments, the energy delivered to one or more filaments to create a channel may be in the range of 0.0067 μJ / μm 3 -0.0400μJ / μm 3 . The energy delivered to one or more filaments can be delivered in pulses of 2 to 12 milliseconds (ms) to provide enough energy to produce a continuous channel for delivery. The characteristics of the channel for efficiently and safely delivering drugs through the skin (e.g., from a patch described herein) can vary between in vitro and in vivo embodiments. For example, in some embodiments, when 2mJ / filament to 12mJ / filament energy is applied to one or more filaments in pulses of 2ms to 16ms, one or more filaments produce a channel. In some embodiments, the energy applied to one or more filaments to produce a channel is 2mJ / filament to 8mJ / filament, 2mJ / filament to 6mJ / filament, or 2mJ / filament to 4mJ / filament. In some embodiments, the duration of the pulse is 2ms to 12ms. In such embodiments, one or more filaments may comprise stainless steel or be substantially formed of stainless steel, and for 2mJ / filament-12mJ / filament, have a volume (V) of 300,000μm 3 (0.0067μJ / μm 3 -0.0400mJ / μm 3 ).

[0129] In some embodiments, one or more filaments arranged in a filament array can create 25 channels / cm in a biological membrane (e.g., skin) to which the one or more filaments are exposed. 2 Up to 500 channels / cm 2 In some embodiments, one or more filaments arranged in a filament array can create 50 channels / cm on the skin exposed by the one or more filaments. 2 Up to 400 channels / cm 2 .

[0130] In some embodiments, the cumulative (or total) depth of all channels formed by one or more filaments is about 2500 μm to 30000 μm per square centimeter of skin exposed to the one or more filaments. In some embodiments, the cumulative or total volume of all channels formed by one or more filaments is about 0.05 mm per square centimeter of skin. 3 Up to 0.35mm 3 .

[0131] In some embodiments, the patches described herein, used with channels created by a microporation device, may have one or more features that enhance optimal drug release and diffusion of non-aggregated peptides into the body.

[0132] In some embodiments, improvements to microporation devices and patches as described herein enable microporation drug delivery systems to effectively and safely provide delivery of non-aggregated peptides in a manner that provides improved bioavailability and / or transferability of non-aggregated peptides from the blood of a subject to the cerebrospinal fluid. Embodiments of the microporation drug delivery systems described herein can provide improved patient compliance and enhanced drug delivery capabilities. Embodiments of the microporation drug delivery systems can also provide a reduced risk of adverse reactions caused by uncontrolled delivery and reduce the development period and cost of drugs for patients. Embodiments of the microporation drug delivery systems also enable patients to painlessly and needle-free self-administer the corresponding drug at a location of the patient's choice, which results in improved compliance and reduced costs (fewer visits to health care professionals). Embodiments of the microporation drug delivery systems as described herein can be used for patients with a wide range of skin types, conditions, etc., where the variation in individual drug delivery results is small.

[0133] Example

[0134] Various embodiments and alternatives are disclosed in further detail in the following examples, which are not in any way intended to limit the scope of the claims.

[0135] Alzheimer's disease. Alzheimer's disease is a chronic disease that develops with aging. Therefore, due to the nature of Alzheimer's disease, drugs may require long-term daily administration. Therefore, IV administration is not desirable because it requires hospitalization or daily visits. Subcutaneous administration can be developed as a self-administering device, such as a pen injector. However, it is believed that it will not be effective for most patients in the long term due to pain and handling issues for the patient. In addition, patients sometimes suffer from needle phobia. Therefore, it is believed that percutaneous microporation will have good compliance and will serve as a long-term home therapy. In this study, a percutaneous microporation device was studied for the non-aggregating peptide p3-Alcβ to enhance the delivery of p3-Alcβ to the brain compared to other modes of administration and to determine the effectiveness of various modes of administration.

[0136] p3-Alcβ. Figure 1 The amino acid sequences of p3-Alcβ1-40, p3-Alcβ1-37, p3-Alcβ9-19, p3-Alcβ1-19, and p3-Alcβ11-19, which are partial peptides of Alcadeinβ, are described. The human p3-Alcβ37 peptide and its partial peptide p3-Alcβ9-19 were synthesized and purified to a purity of greater than 95%, wherein the human p3-Alcβ37 peptide includes the sequence from Val813 to Thr849 of Alcβ described in Hata et al "Alcadein cleavages by amyloidβ-precursor protein (APP)α-andγ-secretasegenerates small peptides, p3-Alcs, indicating Alzheimer'sdisease-relatedγ-secretase dysfunction" J.Biol.Chem.

[2009] 284, 36024-36033; their expected molecular weights were confirmed by mass spectrometry performed at the Peptide Research Institute (Osaka, Japan).

[0137] Animal studies in rats. Oral, nasal, injection and percutaneous microporation containing p3-Alcβ9-19 were tested to determine pharmacokinetics in vivo, such as PK characteristics, bioavailability and drug recovery. In vivo PK studies were performed in CD rats (Charles River Laboratories) according to an IACUC-approved animal protocol. Each dose formulation was prepared before the experiment. Patches were made according to the dry patch manufacturing procedure. Animals for transdermal application were shaved 1 day before the date of administration. Transdermal microporation was implemented using an applicator and a perforator. Then, the patch was placed to cover the perforated area for 1 day or a specified duration. After the last blood draw, the patch was collected and the application site was wiped with paper or cotton to recover the remaining drug on the skin. Blood samples were collected from the tail vein and transferred to K2-EDTA tubes at the specified sampling time. The samples were centrifuged at 4°C and plasma samples were obtained. All samples were stored in a -80°C refrigerator until bioassay.

[0138] Sample preparation for LCMSMS analysis of p3-Alcβ9-19. Keep the plasma samples at room temperature for 30 minutes. Transfer the plasma (50 μL) to a 2 mL Eppendorf tube (Protein LoBind), then add 50 μL of acetonitrile / DMSO (4:1, v / v), and vortex the tube for a few seconds. Centrifuge the sample at 3700 rpm, 4°C for 15 minutes. Place 150 μL of mobile phase A (0.1% formic acid in water) in a new tube, then transfer 50 μL of the supernatant to the tube, and vortex the sample for 5 minutes. Remove the supernatant and filter the solution through a 0.20 μm PTFE membrane into an automatic sampling vial (LCMSMS).

[0139] LCMSMS method of p3-Alcβ9-19. LCMSMS conditions are shown in Tables 1 and 2.

[0140] Table 1

[0141]

[0142]

[0143] Table 2

[0144] parameter condition Ion spray voltage 5500V Air curtain gas 35psi Ion source gas GS1 / GS2 50 / 50psi Collision gas high temperature 650℃ Analyte / monitor ion 1028.390>383.000 / 514.290>318.300 Declustering potential 40V Inlet potential 10V Collision Energy 20V Collision cell exit potential 10V Diverter valve 1.0 to 2.7 min (introduction to MS / MS) Dwell time 150 ms

[0145] Animal studies in mice. The transport of p3-Alcβ9-19 to blood and cerebrospinal fluid (CSF) was tested in wild-type mice (4 months). Hair was removed before the experiment. Percutaneous microporation was applied, and then a patch containing p3-Alcβ9-19 was applied to the perforated area. Mice were anesthetized with 1% isoflurane, CSF was collected from the cisterna magna of mice, blood was collected from the inferior vena cava of mice, and the CSF and blood were collected into tubes containing EDTA and heparin, 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).

[0146] sELISA method development for p3-Alcβ9-19 analysis. Developed a sELISA method for measuring p3-Alcβ9-19 in cerebrospinal fluid (CSF). Polyclonal rabbit antibodies to p3-Alcβ9-19 (C+HRGHQPPPEMA) containing an amino-terminal Cys residue were stimulated and conjugated to bovine thyroglobulin. IgG was purified with an antigen-coupled resin and conjugated to biotin. Horseradish peroxidase-conjugated streptavidin was from Amersham / GE Healthcare (Cat#RPN1051, Little Chalfont, UK), and tetramethylbenzidine (TMB) microporous peroxidase substrate system was from SeraCare Life Sciences Inc. (Cat#5120-0075, Milford, MA, USA). Mice were anesthetized by using 1% isoflurane, and CSF was collected from the cisterna magna of mice as described above, and then the mice were killed.

[0147] To quantify p3-Alc[beta]9-19 in mouse CSF and plasma, samples were diluted with buffer A (PBS containing 1% bovine serum albumin and 0.05% Tween-20). Using polyclonal antibodies, the developed sELISA system quantified p3-Alcβ9-19 in the range of 25-200 pg / ml, with sensitivity equivalent to that of p3-Alcα35-specific (as described in Omori et al., “Increased levels of plasma p3-Alcα35, a major fragment of Alcadeiνα byγ-secretase cleavage, in Alzheimer's disease” J. Alzheimers. Dis. (2014) 39, 861-870), p3-Alcβ37- and p3-Alcβ40-specific sELISA systems (as described in Hata et al. “Decrease in p3-Alcβ37and p3-Alcβ40, products of Alcadeinβ generated byγ-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, a 1:10,000-fold diluted antiserum was used as the capture antibody, and affinity-purified antiserum with antigen-coupled resin and biotin-labeled IgG was used for detection antibody. This sELISA system does not react with p3-Alcβ37. The added 1,000pg / ml p3-Alcβ37 does not compete with antibodies that bind to 0-200pg / ml p3-Alcβ9-19. The sELISA method is used to measure p3-Alcβ9-19 in body fluids, even in the presence of endogenous p3-Alcβ in mice.

[0148] sELISA method for p3-Alcβ9-19 analysis. Rabbit antiserum immunized with p3-Alcβ9-19 as an antigen was coated on a 96-well plate as a capture antibody and incubated at 4°C overnight. On the next day, non-bound antibodies were removed with a wash buffer (0.05% Tween-20 in PBS). Plasma was diluted 200 times and cerebrospinal fluid was diluted 20 times with EIA buffer (1% BSA, 0.05% Tween-20), respectively. The sample solution was added to a 96-well plate and incubated at 4°C overnight. On the next day, after washing with a wash buffer, purified biotinylated IgG dissolved in PBS was added to each well to 1.0 μg / mL, and the mixture was kept at 4°C overnight. On the next day, unreacted antibodies were washed with a wash buffer, streptavidin-HRP (1 / 5000 in PBS) was added, and the reaction was allowed to proceed at 4°C for 6 to 8 hours. After washing with washing buffer, TMB color development substrate was added, and the mixture was reacted at room temperature for 30 minutes under light protection. Color development was stopped by adding 1N H2SO4, and the sample was measured at 450nm by a plate reader.

[0149] Mitochondrial activation in the brain. Monitoring brain mitochondrial function by PET imaging using the [18F]BCPP-EF probe, which can detect mitochondrial complex I activity reflecting neuronal activity in the living brain, confirmed that p3-Alcβ increased neuronal activity in vivo. Mitochondrial dysfunction is often present in the brains of AD patients, and reduced activity in vulnerable brain regions can be detected by PET imaging with [18F]BCPP-EF.

[0150] Percutaneous microporation device. Figure 2A Examples of patches are described; Figure 2BThe patch application after percutaneous microperforation in the skin is shown. The microperforation device consists of (a) an applicator electrically connected to the filament, (b) a perforator having an upper substrate surface and defining a perforation area, and (c) a patch. A filament array having a plurality of filaments is arranged in the perforation area, and each filament is capable of conductively transferring heat energy to form at least one micropore in the tissue membrane via direct contact with the tissue membrane. The applicator provides a predetermined electrical energy to the filament to generate micropores, and applies the patch to at least a portion of the micropores. The patch (100) consists of a backing (200), a matrix (300) containing a composition (302) of p3-Alcβ and ingredients in a dry state, for example, and a release liner (400) that is removed before being applied to the perforated skin. The backing (200) may have an adhesive (201). The matrix (300) may consist of a matrix support (301) and a composition (302) arranged on the matrix support (301). After percutaneous microporation, the patch (100) is applied to the perforated skin after removing the release liner (400). The active ingredients in the composition (302) are dissolved by the interstitial fluid from the microchannels, and then the active ingredients migrate to the microchannels (percutaneous area) and then enter the blood circulation.

[0151] Patch manufacturing. Patches are manufactured based on distribution or tableting procedures known to those skilled in the art. Individual preparations utilize different ingredients as described herein. The patch includes a backing, a matrix, and a release liner, the matrix comprising non-aggregated peptides and ingredients disposed within the matrix, wherein the release liner is configured to be removed before being applied to the skin of the subject. The non-aggregated peptide is p3-Alcβ9-19. In the distribution method, all ingredients are dissolved in a water and ethanol mixed solvent. A predetermined amount of the ingredient solvent is distributed on a non-woven pad in a patch without a release liner. The distributed patch is moved to a drying chamber at 50°C until dry. The dried patch is taken out of the drying chamber and placed at room temperature for 30 minutes. The patch is covered with a release liner and then cut into a specified size. A separate patch is wrapped in an aluminum laminate bag with a desiccant. In the tableting method, all ingredients are mixed and weighed in a specified amount. The weighed powder is transferred to a die punch of a specified size for a compressor. The mold is set on the machine and compressed. Take the thin rectangular tablet and place it on the nonwoven pad in the patch without the release liner. Cover the patch with the release liner and then cut it into the specified size. Wrap the individual patches in an aluminum laminate bag along with the desiccant.

[0152] Example 1. Administration routes of p3-Alcβ

[0153] Non-parenteral administration route. p3-Alcβ9-19 (5 mg / head) was administered orally, nasally, or passively transdermally in rats. No absorption was observed in all administrations. p3-Alcβ9-19 is a hydrophilic peptide and its molecular weight is 1256 Daltons. These physicochemical properties may affect the poor membrane permeability through the gastrointestinal tract, nasal cavity, and stratum corneum.

[0154] Parenteral Routes of Administration. Figure 3A , Figure 3B and Figure 3C Shown are the PK profiles of p3-Alcβ9-19 following intravenous and subcutaneous administration in rats (mean±SE, n=4). Figure 3A Shown is a line graph depicting the PK profile of p3-Alcβ9-19 following intravenous administration (1 mg / body and 2 mg / body) in rats; Figure 3B A line graph depicting the PK profile of p3-Alcβ9-19 following subcutaneous administration (1 mg / body to 10 mg / body) is shown; Figure 3C A logarithmic scale graph depicting the PK profile of p3-Alcβ9-19 is shown. Figure 4A Shown is a line graph depicting the dose response of p3-Alcβ9-19 following intravenous and subcutaneous administration in rats; Figure 4B Shown is a line graph depicting the dose response of p3-Alcβ9-19 following subcutaneous administration in rats (mean±SE, n=4).

[0155] The PK characteristics of intravenous (IV) and subcutaneous (SC) injections showed rapid absorption and rapid elimination in rats. The blood concentration in SC was slightly more persistent than IV, but it disappeared within 1 hour. Dose-dependent increases were observed in both IV and SC injections. The bioavailability in SC may be similar to or slightly higher than IV injection.

[0156] Delivery of p3-Alcβ using percutaneous microporation in rats. Figure 5A Shown is a line graph depicting immediate release (IR) formulations. Figure 5B A line graph depicting the dose response from percutaneous microporation (400 density, 4 mJ / filament) is shown. Tables 3 and 4 describe the formulations and results of this study, respectively.

[0157] Table 3

[0158] Element AARC009R-G1 AARC009R-G2 AARC009R-G3 p3-Alcβ9-19 4mg 8mg 12mg sucrose 0.5mg 0.5mg 0.5mg Methylparaben 0.2mg 0.2mg 0.2mg Propylparaben 0.02mg 0.02mg 0.02mg

[0159] * Transcutaneous area: 0.5cm 2

[0160] Table 4

[0161]

[0162] *aBA (absolute bioavailability) vs. IV (2 mg / body)

[0163] The p3-Alcβ9-19 delivery using percutaneous microporation showed a higher aBA of 545% to 889% compared to IV (2 mg / body). A dose-dependent increase was observed for the percutaneous microporation system. Even compared to IV and SC injections, microporation delivery was the highest. This suggests that the parenteral route may be affected by the in vivo stability of p3-Alcβ9-19, and that microporation delivery can avoid the first-pass effect (metabolism) and maintain the release of p3-Alcβ9-19, rather than injection showing higher bioavailability.

[0164] Example 2. Brain delivery of p3-Alcβ using percutaneous microporation in mice

[0165] The formulations used for the studies are provided in Table 5. Fig. 6A Shown are logarithmic scale graphs depicting changes in blood and central concentrations following percutaneous microporation of an IR formulation containing p3-Alc[beta]9-19 in mice. Figure 6B Shown are logarithmic scale graphs depicting changes in blood and central concentrations following sustained release (SR) formulations containing p3-Alcβ9-19 in mice using percutaneous microporation. Table 6 describes PK parameters in plasma following administration of p3-Alcβ9-19 in mice.

[0166] Table 5

[0167] Element IR preparations* SR preparation* p3-Alcβ9-19 1mg 1mg Lactose monohydrate 1 mg 1mg Disodium citrate sesquihydrate - 2.23mg Methylparaben 0.05mg 0.05mg Propylparaben 0.005mg 0.005mg

[0168] * Transcutaneous area: 0.5cm 2

[0169] Table 6

[0170]

[0171] *BA (relative bioavailability) vs. SC (30 μg / body)

[0172] **Percutaneous microporation: 400 density, 3mJ / filament

[0173] ***Percutaneous microporation: 100 density, 2mJ / filament

[0174] Table 7 describes PK parameters in cerebrospinal fluid (CSF) following administration of p3-Alc[beta]9-19 in mice.

[0175] Table 7

[0176]

[0177] *BA (relative bioavailability) vs. SC (30 μg / body)

[0178] **Percutaneous microporation: 400 density, 3mJ / filament

[0179] ***Percutaneous microporation: 100 density, 2mJ / filament

[0180] Fig. 6A and Figure 6B It was shown that when an IR formulation containing p3-Alcβ9-19 was administered to mice using a transdermal microporation device ( Figure 5A ) or SR preparation ( Figure 5B ), changes in the concentration of p3-Alcβ9-19 in plasma and cerebrospinal fluid (CSF). Tables 2 and 3 show that when an IR preparation containing 1 mg of p3-Alcβ9-19 is administered to mice, the maximum blood concentration (Cmax) is about 600 ng / mL 1 hour after administration, and the blood concentration is maintained at about 300 ng / mL for up to 6 hours. In CSF, >20 ng / mL is shown 3 hours after administration, and a concentration of about 5 ng / mL is detected after 6 hours. This indicates that the transfer rate of p3-Alcβ9-19 from blood to CSF ​​is about 3.5% (BA in CSF / BA in plasma × 100%) of good central transfer. The SF preparation shows sustained plasma concentrations in mice, and the CSF concentration is close to the detection limit.

[0181] Example 3. Pharmacological effects of p3-Alcβ percutaneous microporation in monkeys

[0182] In this example, two patch formulations of the present disclosure are illustrated. It is shown that the use of an IR patch is considered to be preferred for achieving effective enhancement of mitochondrial activity.

[0183] The mitochondrial activation effect of p3-Alcβ9-19 percutaneous microporation was studied in rhesus monkeys. The preparations used for the study are shown in Table 8 below. As a result, [18F]BCPP-EF accumulation, i.e., mitochondrial activation, was observed by PET measurement 1.5 hours after p3-Alcβ9-19 was administered using percutaneous microporation. In addition, activation continued in some areas even after 6.5 hours.

[0184] Table 8

[0185] Element IR preparations* SR preparation* p3-Alcβ9-19 5mg 5mg Lactose monohydrate 1 mg 1mg Disodium Citrate - 4mg Methylparaben 0.1mg 0.1mg Propylparaben 0.01mg 0.01mg

[0186] * Transcutaneous area: 0.5cm 2

[0187] Figure 7Two images of changes in mitochondrial activity in the brain after IR formulations (left) and SR formulations (right) containing p3-Alcβ9-19 using percutaneous microporation in monkeys are shown. Percutaneous microporation applied 400 density and 4mJ / filament for IR formulations and 100 density and 3mJ / filament for SR formulations. An increase in mitochondrial activity (marked with a red rectangle) was found in monkeys, with an immediate response (early) 1 hour after the IR formulation containing p3-Alcβ9-19 was applied, and a slow response (delay) 6.5 hours after the SR formulation containing p3-Alcβ9-19 was applied compared to the control (vehicle).

[0188] Figure 8 Shows a description from Figure 7 Two bar graphs of the intensity of Figure 8 In the text, 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) is the applied IR formulation, Figure 8 (Bottom) is the SR formulation applied. Figure 8 In (top), the intensity at 1 hour appears higher in all regions compared to the control (vehicle), but decreases in intensity at 6.5 hours in all regions except the core shell. Figure 8 (bottom), the intensity gradually increases in all regions.

[0189] Fig. 9 A bar graph depicting the percentage increase in mitochondrial activity in various regions of the brain 1 hour after administration of an IR formulation containing p3-Alcβ9-19 (5 mg / body or 10 mg / body equals 0.5 mg / kg or 1.0 mg / kg, respectively) is shown. Except in the frontal lobe and nucleus and shell regions, the percentage increase for 10 mg / body was higher than for 5 mg / body.

[0190] Example 4. Controlling PK Profiles Using IR Formulations

[0191] Table 9 describes the IR formulations used for the study, which contain p3-Alcβ9-19, an enhancer (ie, non-reducing sugar: sucrose), an antimicrobial (a combination of methylparaben and propylparaben or benzalkonium chloride). Transdermal microporation with different combinations of density and energy levels was applied to study PK changes.

[0192] Table 9

[0193] Element AALC004R AALC005R AALC006R AALC029R AALC030R p3-Alcβ9-19 4mg 4mg 8mg 5mg 5mg sucrose 0.5mg 0.5mg 1mg 1mg 0.5mg Methylparaben 0.2mg 0.2mg 0.2mg - - Propylparaben 0.02mg 0.02mg 0.02mg - - Benzalkonium chloride - - - 0.025mg 0.025mg

[0194] * Transcutaneous area: 0.5cm 2

[0195] Fig. 10A A line graph depicting the PK profile of non-reducing sugar formulations and p3-Alcβ concentration over time at different intensities of 4 mJ / filament is shown; Fig. 10B A logarithmic scale graph depicting the change in p3-Alcβ concentration over time is shown.

[0196] Table 10 shows the results of PK profile types using IR formulations containing p3-Alcβ9-19 under different microporation conditions, including Fig. 10A Results. In an immediate release (IR) formulation, the PK profile can be controlled by changing the percutaneous microporation conditions. No absorption was observed without percutaneous microporation. The 200 density at 4mJ / filament and the 400 density at 2mJ / filament to 4mJ / filament showed the immediate PK profile of p3-Alcβ. On the other hand, the microporation conditions (50≤density≤200density and <4mJ / filament) provided a sustained PK profile for p3-Alcβ.

[0197] Table 10

[0198] 2mJ / filament 3mJ / filament 4mJ / filament 50 Density - - Sustained PK characteristics 100 Density Sustained PK characteristics - Sustained PK characteristics 200 Density Sustained PK characteristics Sustained PK characteristics Real-time PK features 400 Density Real-time PK features - Real-time PK features

[0199] Example 5. Controlling PK characteristics using SR formulations

[0200] Table 11 describes the SR formulation used for the study, which contains p3-Alcβ9-19, enhancer (ie, non-reducing sugar: sucrose), PK modifier (disodium citrate sesquihydrate), antimicrobial (methylparaben / propylparaben).

[0201] Table 11

[0202] Element AALC011R-G4 AALC011R-G5 AALC011R-G6 p3-Alcβ9-19 8mg 8mg 8mg sucrose 0.5mg 0.5mg 0.5mg Disodium citrate sesquihydrate 1 mg 2mg 4mg Methylparaben 0.2mg 0.2mg 0.2mg Propylparaben 0.02mg 0.02mg 0.02mg

[0203] * Transcutaneous area: 0.5cm 2

[0204] Table 12 provides an overview of the types of PK profiles of SR formulations containing p3-Alcβ9-19 and different amounts of PK modifiers. PK modifiers (i.e., containing organic acids, salts of organic acids, or combinations thereof) were added to sustained release (SR) formulations. Sustained PK profiles of p3-Alcβ9-19 were obtained with 4 mg of disodium citrate sesquihydrate at 200 density, 4 mJ / filament.

[0205] Table 12

[0206] AALC011R-G4 AALC011R-G5 AALC011R-G6 200d,4mJ Real-time PK features Real-time PK features Sustained PK characteristics

[0207] Table 13 describes the SR formulations used for the study including p3-Alcβ9-19, enhancer (non-reducing sugar: sucrose), PK modifier (disodium citrate sesquihydrate), antimicrobial (methylparaben / propylparaben). Percutaneous microporation was applied with a PK modifier (4 mg disodium citrate sesquihydrate) at different combinations of density and energy levels.

[0208] Table 13

[0209] Element AALC007R AALC013R and AALC014R p3-Alcβ9-19 8mg 8mg sucrose 1mg 0.5mg Disodium citrate sesquihydrate 4 mg 4mg Methylparaben 0.2mg 0.2mg Propylparaben 0.02mg 0.02mg

[0210] * Transcutaneous area: 0.5cm 2

[0211] Table 14 summarizes the PK profiles using different microporation conditions with PK modifiers.Sustained release (SR) formulations containing disodium citrate sesquihydrate showed sustained PK profiles with 4 mg disodium citrate sesquihydrate at densities below 400 and 4 mJ / filament.

[0212] Table 14

[0213] 2mJ 3mJ 4mJ 100 Density Sustained PK characteristics - Sustained PK characteristics 200 Density Sustained PK characteristics - Sustained PK characteristics 400 Density - - Sustained PK characteristics

[0214] Table 15 summarizes the PK profiles of IR and SR formulation types containing p3-Alcβ controlled using percutaneous microporation. Immediate PK profiles were provided using IR type formulations (without PK modifiers) under microporation conditions of less than 200 density and 2-4 mJ / filament. Sustained PK profiles were obtained with IR formulations of 50≤density≤200 density and <4 mJ / filament and SR formulations containing PK modifiers of less than 400 density and 4 mJ / filament.

[0215] Table 15

[0216]

[0217] Example 6. Enhancing p3-Alcβ delivery using enhancers

[0218] Table 16 describes dry patch formulations to study the effect of enhancers (sugars) on the delivery of p3-Alcβ9-19 using transdermal microporation. Enhancers were selected from non-reducing sugars (sucrose, trehalose, D-mannitol and D-sorbitol) and reducing sugars (lactose and maltose).

[0219] Table 16

[0220]

[0221] * Transcutaneous area: 0.5cm 2

[0222] Fig.11 The PK profiles are shown in the presence of non-reducing sugars or reducing sugars. Reducing sugars (lactose and maltose) showed higher delivery than non-reducing sugars. Table 17 describes the PK profiles from Fig.11 Results. All non-reducing or reducing sugars increased p3-Alcβ absorption compared to IV injection. The aBA of non-reducing sugars ranged from 194.3% to 547.3%. In particular, reducing sugars (such as maltose and lactose) showed significant increases in p3-Alcβ9-19 (aBA: 2923.3% and 4121.7%) using percutaneous microporation.

[0223] Table 17

[0224]

[0225] *aBA (absolute bioavailability) vs IV (2 mg / body)

[0226] **Percutaneous microporation: 400 density, 4mJ / filament

[0227] Example 7. Effect of Antimicrobial Agents on p3-Alcβ Delivery

[0228] Antimicrobial agents are added as preservatives or antimicrobial effects to avoid the risk of infection. Table 18 describes examples of formulations with antimicrobial agents.

[0229] Table 18

[0230]

[0231] * Transcutaneous area: 0.5cm 2

[0232] Table 19 describes the PK parameters after application of the formulations in rats using transdermal microporation (400 density, 4 mJ / filament). It was shown that reducing sugars acted together with the antimicrobial agents (a combination of methylparaben and propylparaben, or a benzalkonium chloride formulation).

[0233] Table 19

[0234]

[0235] *aBA (absolute bioavailability) vs. IV (2 mg / body)

[0236] **Percutaneous microporation: 400 density, 4mJ / filament

[0237] Example 8. Immediate and sustained PK delivery using reducing sugar formulations

[0238] Table 20 describes IR (without PK modifier) ​​and SR (with PK modifier) ​​formulations containing reducing sugars (lactose) and p3-Alcβ9-19 to control PK characteristics.

[0239] Table 20

[0240]

[0241] * Transcutaneous area: 0.5cm 2

[0242] Fig. 12A An example of a line graph depicting the PK profiles of IR and SR formulations containing a reducing sugar (lactose) and p3-Alcβ formulations in rats is shown; Fig. 12B A logarithmic scale graph depicting the change in p3-Alcβ concentration over time is shown. Plasma concentrations of p3-Alcβ9-19 showed a typical immediate and sustained PK profile following administration of IR and SR formulations containing reducing sugars using transdermal microporation.

[0243] Table 21 describes the results of PK parameters after administration of formulations using percutaneous microporation in rats. IR and SR formulations containing reducing sugars each showed higher BA compared to non-reducing sugar formulations.

[0244] Table 21

[0245]

[0246] *aBA (absolute bioavailability) vs. IV (2 mg / body)

[0247] **Percutaneous microporation: 400 density, 4mJ / filament

[0248] ***Percutaneous microporation: 100 density, 3mJ / filament

[0249] Example 9. Effect of Density on p3-Alcβ Delivery Using Reducing Sugar Formulations

[0250] Table 22 describes formulations tested for p3-Alcβ9-19 delivery at various microporation densities.

[0251] Table 22

[0252] Element AALC007R p3-Alcβ9-19 5mg Lactose monohydrate 1mg Benzalkonium chloride 0.025mg

[0253] * Transcutaneous area: 0.5cm 2

[0254] Fig.13The drug residue at 24 hours after the application of the reducing sugar formulation using percutaneous microporation (4 mJ / filament) in rats is shown. p3-Alcβ9-19 was recovered by about 100% without microporation. The drug recovery rate decreased with the increase of percutaneous microporation density. At a density of 200 to 400, 10% to 20% of p3-Alcβ9-19 was recovered. Density exceeding 200 showed effective delivery of p3-Alcβ9-19.

[0255] Example 10. Optimization of enhancer content in reducing sugar preparations

[0256] Table 23 describes the formulations used to study the effect of enhancer content in p3-A1c[beta] transdermal microporation.

[0257] Table 23

[0258]

[0259] Fig.14 A line graph depicting the PK profile of rats with different enhancer contents (reducing sugar, lactose) is shown. Table 24 describes the Fig.14 PK parameters. Fig.15 A line graph comparing AUC versus lactose content is shown.

[0260] Table 24

[0261]

[0262] *aBA (absolute bioavailability) vs. IV (2 mg / body)

[0263] **Percutaneous microporation: 400 density, 4mJ / filament

[0264] The plasma concentration of p3-Alcβ9-19 increased with the increase of enhancer content (G1-G5). No enhancement was observed in the absence of enhancer (G7). The ratio between p3-Alcβ and enhancer may be related to the enhancer factor.

[0265] Example 11. Dose dependence of p3-Alcβ delivery using IR and SR formulations

[0266] Immediate delivery system. Tables 25 and 26 describe the dose dependence of IR formulations containing p3-Alcβ using transdermal microporation.

[0267] Table 25

[0268]

[0269] Table 26

[0270]

[0271] Table 27 shows the PK parameters of immediate delivery of different p3-Alcβ9-19 doses in rats using transdermal microporation (400 density, 4 mJ / filament).

[0272] Table 27

[0273]

[0274] *aBA (absolute bioavailability) vs. IV (2 mg / body)

[0275] **Percutaneous microporation: 400 density, 4mJ / filament

[0276] Fig.16 The relationship between p3-Alcβ9-19 dose and AUC is shown.A linear dose dependency in immediate delivery was observed up to 50 mg of p3-Alcβ9-19 (test dose).

[0277] Sustained Delivery System. Tables 28 and 29 describe the dose dependence of SR formulations containing p3-Alcβ using transdermal microporation assay.

[0278] Table 28

[0279]

[0280] Table 29

[0281]

[0282] Table 30 shows the PK parameters of sustained delivery of different p3-Alcβ9-19 doses in rats using transdermal microporation (100 density, 3 mJ / filament).

[0283] Table 30

[0284]

[0285] *aBA (absolute bioavailability) vs. IV (2 mg / body)

[0286] **Percutaneous microporation: 100 density, 3mJ / filament

[0287] Fig.17 The relationship between p3-Alcβ9-19 dose and AUC is shown.A linear dose dependence in sustained delivery was observed up to 25 mg of p3-Alcβ9-19 (test dose).

[0288] Example 13. Optimized patch formulation for p3-Alcβ delivery

[0289] Tables 31 and 32 describe the optimized IR formulations for immediate delivery and patch materials.

[0290] Table 31

[0291]

[0292] Table 32

[0293]

[0294] Fig.18 A line graph depicting the PK profile of an optimized formulation containing p3-Alcβ9-19 for immediate delivery in rats is shown. Table 33 describes Fig.18 Results. p3-Alcβ9-19 was efficiently delivered from the patch and approximately 2000% of the aBA was obtained compared to IV injection. Tmax was approximately 1 hour.

[0295] Table 33

[0296]

[0297] *aBA (absolute bioavailability) vs. IV (2 mg / body)

[0298] **Percutaneous microporation: 400 density, 4mJ / filament

[0299] Fig.19A The relationship between p3-Alcβ9-19 dose and AUC is described. Fig.19B The relationship between p3-Alcβ9-19 dose and Cmax is described. AUC and Cmax increased with increasing p3-Alcβ9-19 dose-dependently.

[0300] Physicochemical properties of p3-Alcβ9-19 patch. The pH and water content of the test preparations. Tables 34 to 36 describe the results. The pH was measured after the patch was mixed in water (USP, pH 5-7). The pH of the preparation was about 4.9 (the acceptable range of pH was set at 4 to 9). After the patch was mixed in anhydrous methanol, the water content in the patch was measured by a volumetric Karl Fisher instrument. The initial water content was approximately less than 2%. Dissolution was tested using device 5 (paddle and disc). The patch was placed in 500 mL phosphate buffered saline (USP) at a speed of 50 rpm and a temperature of 32°C. The drug was measured by RP-HPLC. The drug was released immediately within 5-10 minutes.

[0301] Table 34

[0302]

[0303] Table 35

[0304]

[0305] Table 36

[0306]

[0307] Preliminary stability of p3-Alcβ9-19 patch. The preliminary stability of the formulations was evaluated for 3 months at 25°C / 60% RH and 40°C / 75% RH. Tables 37 to 39 describe the results of 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.

[0308] Table 37

[0309]

[0310] Table 38

[0311]

[0312] Table 39

[0313]

[0314] Table 40 summarizes the results of the investigation of the route of administration of p3-Alcβ. Transdermal microporation for delivery of p3-Alcβ9-19 is an ideal administration device, and the patch formulation shows good room temperature storage stability. This is important for patient compliance and the supply chain.

[0315] Table 40

[0316] Application PK characteristics Bioavailability (%)* oral No absorption 0 nasal cavity No absorption 0 Passive percutaneous No absorption 0 Intravenous injection Bolus absorption 100 Subcutaneous injection Mass absorption 120–275 Percutaneous microporation Instant absorption or continuous absorption –4800

[0317] *Compared to IV (2 mg)

[0318] In addition, although the foregoing has been described in detail by way of illustration and example for the purpose of clarity and understanding, it will be appreciated by those skilled in the art that many and various modifications may be made without departing from the spirit of the present disclosure. Therefore, it should be clearly understood that the forms disclosed herein are illustrative only and are not intended to limit the scope of the present disclosure, but also encompass all modifications and substitutions that come with the true scope and spirit of the present disclosure.

Claims

1. A patch for delivering a non-aggregated peptide to a subject, the patch comprising: Backing; a matrix comprising non-aggregated peptides disposed within the matrix; and Release liner, Wherein the release liner is configured to be removed prior to application to the skin of a subject.

2. The patch of 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 derivatives thereof. The patch of claim 2 , wherein the p3-Alcβ is p3-Alcβ9-19 or a derivative thereof.

5. The patch of any one of claims 1 to 4, wherein the amount of the non-aggregated peptide in the matrix ranges from about 0.01 mg / cm 2 To about 200mg / cm 2 .

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

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

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

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

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

02.

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

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

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

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

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

17. The patch of 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 of 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 of any one of claims 1 to 18, wherein the substrate comprises at least one laminate of fibers, films, or a combination thereof.

20. The patch of claim 19, wherein the substrate comprises a laminate of film and at least one fiber.

21. The patch of claim 19 or 20, wherein the at least one fiber is a nonwoven fiber.

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

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

24. The patch of any one of claims 1 to 23, wherein the matrix has a viscosity of less than about 10 mg / cm 2 water holding capacity.

25. A device for delivering a non-aggregated peptide to a subject through a plurality of microchannels, the device comprising: A patch according to any one of claims 1 to 24; a perforator comprising an array of electrically conductive filaments; and An applicator is electrically connected to the conductive filaments and is configured to provide a predetermined electrical energy to the array of conductive filaments for generating a plurality of micropores in a micropore region of the subject's skin by heating the filaments.

26. The device of claim 25, wherein the device produces from about 25 to about 500 microchannels / cm 2 .

27. The device of claim 25 or 26, wherein the device has a perforation energy of about 2 to about 6 mJ / filament, 4 mJ / filament, or 8 mJ / filament.

28. The device of any one of claims 25 to 27, wherein the perforator is configured to open at least one channel in the skin of the subject and has a diameter of about 0.25 cm. 2 To about 4cm 2 area.

29. The apparatus of claim 28, wherein the perforator is configured to open at least one channel in the subject's skin and has a diameter of about 0.1 cm. 2 、About 0.25cm 2 、About 0.65cm 2 or about 1.0cm 2 area.

30. The device of any one of claims 28 or 29, wherein the perforator is configured to open at least one channel in the skin of the subject and has a diameter of less than about 1.0 cm. 2 、About 0.5cm 2 or about 0.25cm 2 area.

31. The device of claim 30, wherein the at least one channel is one or more micropores, wherein the perforator is configured to create the one or more micropores in about 0.5% to about 12.5% ​​of the total perforated area.

32. The device of any one of claims 25 to 31, wherein the perforator is a microneedle, laser or radiofrequency perforator configured to create one or more micropores in the skin of the subject.

33. The device of any one of claims 25 to 32, wherein the perforator is configured to create at least 50 holes in the skin of the subject.

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

35. The device of 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, the method comprising: opening at least one channel in the skin of the subject; and A patch as claimed in any one of claims 1 to 24 is applied to the skin of the subject.

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

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

39. The method of any one of claims 36 to 38, wherein opening at least one passage in the subject's skin comprises applying a percutaneous microporation device to the subject's skin.

40. The method of claim 39, wherein the percutaneous microporation device is thermal tissue cauterization performed by using a filament array having a plurality of filaments disposed in the subject's skin, wherein each filament is capable of conductively transferring thermal energy via direct contact with a tissue membrane to form a plurality of micropores in a microporous region of the tissue membrane.

41. The method of claim 36 or 40, wherein the percutaneous microporation device creates from about 25 to about 500 microchannels / cm 2 .

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

43. The method of any one of claims 36 to 42, wherein at least one channel is opened in the subject's skin with a length of about 0.25 cm 2 To about 4cm 2 area.

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

45. The method of any one of claims 36 to 44, wherein applying the patch increases neuronal activity in the subject.

46. ​​The method of any one of claims 36 to 44, wherein applying the patch increases mitochondrial activity in the subject's brain.

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

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

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

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

51. The method of claim 43, wherein the at least one channel opened in the subject's skin has a length of about 0.1 cm 2 、About 0.25cm 2 、About 0.65cm 2 or about 1.0cm 2 area.

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

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

54. The system of any one of claims 52 or 53, wherein the percutaneous microporation device creates from about 25 to about 500 microchannels / cm 2 .

55. The system of any one of claims 52 or 54, wherein the percutaneous microporation device has a perforation energy of about 2 mJ / filament to about 10 mJ / filament.

56. The system of any one of claims 52 to 55, wherein the at least one channel opened in the subject's skin has a width of about 0.25 cm 2 To about 4cm 2 area.

57. The system of any one of claims 52 to 56, wherein the one or more micropores comprise between about 0.5% and about 12.5% ​​of the total perforated area.

58. The system of claim 52, wherein the transdermal microporation device is a microneedle, laser, or radiofrequency device capable of creating one or more micropores in the subject's skin.

59. The system of any one of claims 52 to 58, wherein the transcutaneous microporation device creates at least 50 holes in the skin of the subject.

60. The system of any one of claims 52 to 59, wherein the patch further comprises a drug pellet.

61. The system of claim 56, wherein the substrate is configured to open at least one channel in the subject's skin and has a thickness of about 0.1 cm 2 、About 0.25cm 2 、About 0.65cm 2 or about 1.0cm 2 area.

Images