Microneedle array patch for expression of therapeutics

Abstract

Microneedle patches or microneedle array patches (MAPs) offer a convenient method to intradermally deliver drugs and vaccines. Embodiments include MAPs that overcome the limitations of conventional MAPs. In aspects, the MAPs include (a) a base, (b) an adhesive region and (c) a plurality of needles on a surface of the base. The needles can comprise a medicament (e.g., a therapeutic or vaccine). In aspects, the medicament is a circular RNA vaccine that elicits an immune response against SARSCoV-2.

Description

PATENT EX1-005WOMICRONEEDLE ARRAY PATCH FOR EXPRESSION OF THERAPEUTICSRELATED APPLICATIONS

[0001] The present application claims priority to U. S. Patent Application No.63 / 760,524 filed February 19, 2025 and U. S. Patent Application No. 63 / 951,329 filed December 30, 2025. The contents of the aforementioned applications are incorporated by reference.FIELD OF THE INVENTION

[0002] The invention relates to medical devices, and more specifically, it relates to a method and device for administering a medicament or vaccine into the skin.BACKGROUND

[0003] Drug delivery refers to approaches, formulations, manufacturing techniques, storage systems, and technologies involved in transporting a pharmaceutical compound to its target site to achieve a desired therapeutic effect. Principles related to drug preparation, route of administration, site-specific targeting, metabolism, and toxicity are used to optimize efficacy and safety, and to improve patient convenience and compliance. Drug delivery is aimed at altering a drug's pharmacokinetics and specificity by formulating it with different excipients, drug carriers, and medical devices. There is additional emphasis on increasing the bioavailability and duration of action of a drug to improve therapeutic outcomes. Some research has also been focused on improving safety for the person administering the medication. For example, several types of microneedle patches have been developed for administering vaccines and other medications to reduce the risk of needlestick injury.

[0004] Microneedle patches or microneedle array patches (MAPs) offer a convenient method to intradermally deliver drugs and vaccines. MAPs are a type of transdermal patch that has been embedded with an array of tiny needles to deliver the drug formulation. The patches of microneedles are placed against the skin. The needles puncture the skin to facilitate drug delivery. A microneedle patch can include an adhesive layer that sticks to the skin and needles that puncture the skin to facilitate drug delivery. Microneedle patches usually contain the drug formulation meant to enter thePATENT EX1-005WObody. Alternatively, they are used in tandem with drug-containing transdermal patches.

[0005] There are many benefits to the use of MAPs rather than conventional methods of drug administration. For example, transdermal administration allows rapid drug delivery that bypasses that gastrointestinal (Gl) tract thus eliminating first-pass metabolism. They can be stored at ambient temperature, thus eliminating the need for a cold supply chain. Moreover, there is no need for reconstitution. MAPs can be applied to the skin by a layperson or patient, so there is no need for a healthcare provider. They typically have a short patch wear time (e.g., five minutes) with minimal discomfort. Moreover, recent studies have demonstrated that majority of subjects preferred patch to intramuscular (IM) injection.

[0006] There is a need for improved MAPs and methods of using them. The devices and methods should be safe and effective with minimal side effects. Aspects of the present disclosure fulfill these needs and provide further related advantages as described in the following summary.SUMMARY OF THE INVENTION

[0007] The inventions described and claimed herein have many attributes and embodiments including, but not limited to, those set forth or described or referenced in this brief summary. The inventions described and claimed herein are not limited to, or by, the features or embodiments identified in this summary, which is included for purposes of illustration only and not restriction.

[0008] Embodiments include microneedle array patches (MAPs) for administering a formulation with an active agent. In aspects, the microneedle array patch (MAP) includes (a) a base, (b) an adhesive region and (c) a plurality of needles on a surface of the base. The needles can comprise a medicament (e.g., a therapeutic or vaccine).

[0009] In aspects, the active agent is a circular RNA (circRNA). In aspects, the circRNA encodes at least one antigenic peptide or protein of a virus that induces an efficient antigen-specific immune responses against the encoded antigenic peptide or protein.

[0010] In embodiments, the MAPs described herein include solid microneedles.PATENT EX1-005WOAlternatively, the MAPs use coated microneedles, hollow microneedles or dissolvable microneedles. In aspects, the microneedles are hydrogel-forming microneedles.

[0011] The active agent can be a small molecule therapeutic (e.g., aspirin, acetaminophen, antihistamines, ibuprofen or penicillin). Alternatively, the active agent can be a large molecule therapeutic (e.g., a protein, a peptide, a nucleotide sequence, a monoclonal antibody, a vaccine or a polysaccharide).

[0012] Aspects include a reusable kinetic applicator, with separate individually packaged coated patch assemblies. Aspects also include a single-use applicator.

[0013] Embodiments also include a device and methods for preparing a delivery site on the skin to enhance the delivery of an active agent through the stratum corneum of the skin to a sufficient depth so that the active agent can be absorbed and utilized by the body.

[0014] Embodiments also include a method of intradermal administration of a vaccine to a subject. The method can include steps of (a) providing a microneedle array patch (MAP) and (b) applying the MAP to an area of skin of the subject. In aspects, the MAP comprises a plurality of needles on a surface wherein each needle comprises a vaccine composition.

[0015] Embodiments also include a microneedle array patch that includes (a) a backing substrate, (b) a plurality of microneedles extending from the backing substrate, each microneedle having a length of 300 - 1000 pm and (c) a coating disposed on at least a portion of the microneedles. The coating can include (i) a lipid nanoparticle encapsulating a circular RNA, (ii) sucrose, and (iii) a polymer selected from polyvinyl alcohol (PVA) and polyvinylpyrrolidone (PVP). The circular RNA can include: (A) an internal ribosome entry site (IRES), (B) an open reading frame encoding a therapeutic protein and (C) a covalently closed circular backbone.

[0016] Embodiments also include a pharmaceutical formulation for coating a microneedle array patch, that includes (a) a lipid nanoparticle encapsulating a circular RNA comprising an IRES and an open reading frame encoding a therapeutic protein,PATENT EX1-005WO(b) sucrose, (c) a polymer selected from PVA and PVP and (d) water.BRIEF DESCRIPTION OF THE FIGURES

[0017] FIG. 1 A depicts a microneedle array according to embodiments.

[0018] FIG. 1B depicts dimensions of individual microneedles, according to embodiments.

[0019] FIG. 2A depicts a top view of a microneedle array patch (MAP) and applicator insert, according to embodiments.

[0020] FIG. 2B depicts a bottom view of a microneedle array patch (MAP) and applicator insert, according to embodiments.

[0021] FIG. 3 depicts a microneedle array patch (MAP) kinetic applicator and an adhesive patch containing microneedle array, according to embodiments.

[0022] FIG. 4 shows images of microneedles coated with Formulation H (polyvinyl alcohol) (magnified 100x).Definitions

[0023] Reference in this specification to "one embodiment / aspect" or "an embodiment / aspect" means that a particular feature, structure, or characteristic described in connection with the embodiment / aspect is included in at least one embodiment / aspect of the disclosure. The use of the phrase "in one embodiment / aspect" or "in another embodiment / aspect" in various places in the specification are not necessarily all referring to the same embodiment / aspect, nor are separate or alternative embodiments / aspects mutually exclusive of other embodiments / aspects. Moreover, various features are described which may be exhibited by some embodiments / aspects and not by others. Similarly, various requirements are described which may be requirements for some embodiments / aspects but not other embodiments / aspects. Embodiment and aspect can be in certain instances be used interchangeably.

[0024] The terms used in this specification generally have their ordinary meanings in thePATENT EX1-005WOart, within the context of the disclosure, and in the specific context where each term is used. Certain terms that are used to describe the disclosure are discussed below, or elsewhere in the specification, to provide additional guidance to the practitioner regarding the description of the disclosure. It will be appreciated that the same thing can be said in more than one way.

[0025] Consequently, alternative language and synonyms may be used for any one or more of the terms discussed herein. Nor is any special significance to be placed upon whether or not a term is elaborated or discussed herein. Synonyms for certain terms are provided. A recital of one or more synonyms does not exclude the use of other synonyms. The use of examples anywhere in this specification including examples of any terms discussed herein is illustrative only, and is not intended to further limit the scope and meaning of the disclosure or of any exemplified term. Likewise, the disclosure is not limited to various embodiments given in this specification.

[0026] Without intent to further limit the scope of the disclosure, examples of instruments, apparatus, methods and their related results according to the embodiments of the present disclosure are given below. Note that titles or subtitles may be used in the examples for convenience of a reader, which in no way should limit the scope of the disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. In the case of conflict, the present document, including definitions, will control.

[0027] The term “active agent” or “active ingredient” refers to a substance, compound, or molecule, which is biologically active or otherwise, induces a biological or physiological effect on a subject to which it is administered to. In other words, “active agent” or “active ingredient” refers to a component or components of a composition to which the whole or part of the effect of the composition is attributed. An active agent can be a primary active agent, or in other words, the component(s) of a composition to which the whole or part of the effect of the composition is attributed. An active agent can be a secondary agent, or in other words, the component(s) of a composition to which anPATENT EX1-005WOadditional part and / or other effect of the composition is attributed.

[0028] The term “hydrogel” refers to a biphasic material, a mixture of porous and permeable solids and at least 10% of water or other interstitial fluid. The solid phase is a water insoluble three-dimensional network of polymers, having absorbed a large amount of water or biological fluids. Hydrogels have several applications, especially in the biomedical area, such as in hydrogel dressing. Many hydrogels are synthetic, but some are derived from natural materials. Hydrogels are typically used in tissue engineering, wound dressings, lubricants, contact lenses and drug delivery.

[0029] The term “small molecule” or “micromolecule” refers to a low molecular weight (< 1000 daltons) organic compound that may regulate a biological process, with a size on the order of 1 nm. Larger structures such as nucleic acids and proteins, and many polysaccharides are not small molecules, although their constituent monomers (e.g., ribo- or deoxyribonucleotides, amino acids, and monosaccharides, respectively) are often considered small molecules.

[0030] In some embodiments, drugs that are of a large molecular weight may be delivered intradermally. Increasing molecular weight of a drug typically causes a decrease in unassisted transdermal delivery. Examples of such large molecules include proteins, peptides, nucleotide sequences, monoclonal antibodies, vaccines, polysaccharides, such as heparin, and antibiotics, such as ceftriaxone. Examples of suitable vaccines include flu vaccine, Lyme disease vaccine, rabies vaccine, measles vaccine, mumps vaccine, chicken pox vaccine, small pox vaccine, hepatitis vaccine, pertussis vaccine, rubella vaccine, diphtheria vaccine, encephalitis vaccine, yellow fever vaccine, recombinant protein vaccine, DNA vaccines, polio vaccine, therapeutic cancer vaccine, herpes vaccine, pneumococcal vaccine, meningitis vaccine, whooping cough vaccine, tetanus vaccine, typhoid fever vaccine, cholera vaccine, tuberculosis vaccine, and combinations thereof. The term “vaccine” thus includes, without limitation, antigens in the forms of proteins, polysaccharides, oligosaccharides, or weakened or killed viruses. Additional examples of suitable vaccines and vaccine adjuvants are described in the art (see. e.g., U. S. 2004 / 0049150).PATENT EX1-005WO

[0031] The term “subject” or "patient" refers to any single animal, more preferably a mammal (including such non-human animals as, for example, dogs, cats, horses, rabbits, zoo animals, cows, pigs, sheep, and non-human primates) for which treatment is desired. Most preferably, the patient herein is a human.

[0032] The term “pharmaceutically acceptable carrier” as used herein refers to any and all solvents, dispersion media, coatings, isotonic and absorption delaying agents, and the like, that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. The compositions may also contain other active compounds providing supplemental, additional, or enhanced therapeutic functions.

[0033] The term “pharmaceutically acceptable composition” as used herein refers to a composition comprising at least one compound as disclosed herein formulated together with one or more pharmaceutically acceptable carriers.

[0034] The term “circular RNA” or “circRNA” refers to a type of single-stranded RNA which, unlike linear RNA, comprises a covalently closed continuous loop. circRNAs occur naturally in mammalian cells, and play important roles in various biological processes. circRNAs innately possess greater stability and resistance to intra- and extracellular RNAses than mRNAs, making them attractive candidates for delivery of key payloads where long-lasting expression is necessary. Recently, there has been an interest in using recombinant circRNAs to express a protein of interest, in vitro or in vivo. Introduction of an internal ribosome entry sequence (IRES) into a circular RNA allows translation of a protein encoded by a circRNA. However, IRES elements that exist in nature may or may not support translation from engineered circular RNAs, as IRES elements are often evolved in the context of linear RNA genomes.

[0035] An "infectious" virus or viral particle is one that comprises a polynucleotide component that it is capable of delivering into a cell for which the viral species is tropic. The term does not necessarily imply any replication capacity of the virus. As used herein, an "infectious" virus or viral particle is one that can access a target cell, can infect a target cell, and can express a heterologous nucleic acid in a target cell. Thus,PATENT EX1-005WO"infectivity" refers to the ability of a viral particle to access a target cell, infect a target cell, and express a heterologous nucleic acid in a target cell. Infectivity can refer to in vitro infectivity or in vivo infectivity. Assays for counting infectious viral particles are described elsewhere in this disclosure and in the art. Viral infectivity can be expressed as the ratio of infectious viral particles to total viral particles. Total viral particles can be expressed as the number of viral genome copies. The ability of a viral particle to express a heterologous nucleic acid in a cell can be referred to as "transduction." The ability of a viral particle to express a heterologous nucleic acid in a cell can be assayed using a number of techniques, including assessment of a marker gene, such as a green fluorescent protein (GFP) assay (e.g., where the virus comprises a nucleotide sequence encoding GFP), where GFP is produced in a cell infected with the viral particle and is detected and / or measured; or the measurement of a produced protein, for example by an enzyme-linked immunosorbent assay (ELISA).

[0036] The term “antigen” as used herein will be recognized and understood by the person of ordinary skill in the art, and is for example intended to refer to a substance which may be recognized by the immune system, preferably by the adaptive immune system, and is capable of triggering an antigen-specific immune response, e.g. by formation of antibodies and / or antigen-specific T cells as part of an adaptive immune response. Typically, an antigen may be or may comprise a peptide or protein which may be presented by the MHC to T-cells.

[0037] The term “nanoparticle” refers to a particle of matter, generally between about 1 and 100 nanometers (nm) in diameter. As used herein, a nanoparticle can refer to a liposome, virus, viral vector or other viral particle.

[0038] The term “liposome” refers to a spherical vesicle having at least one lipid bilayer (i.e. an aqueous solution core surrounded by a hydrophobic membrane). Liposomes can be prepared by disrupting biological membranes (such as by sonication).Liposomes are formed when phospholipids and their derivatives are dispersed in water. Upon dispersion in water the phospholipids form closed vesicles called “liposomes,” which are characterized by lipid bilayers encapsulating an aqueous core. VariousPATENT EX1-005WOliposomes have been used as carriers for entrapped therapeutic agents, such as drugs, enzymes and genetic sequences for use in medical science, in pharmaceutical science and in biochemistry. Specific uses include delivery of nutrients and pharmaceutical drugs, such as lipid nanoparticles in mRNA vaccines and DNA vaccines. Liposomes can be modified by the incorporation of polyethylene glycol or other hydrophilic polymers (e.g., a PEG liposome where one or more of the constituent lipids is modified by attachment of PEG). Liposomes can also be modified to target particular cell types by incorporating targeting factors (e.g., “targeting ligands”) for particular cell types. Examples include asialoglycoprotein, folate, transferrin, antibodies, etc.

[0039] The term “exosome” refers to a membrane-bound extracellular vesicles that are produced in the endosomal compartment of most eukaryotic cells. In multicellular organisms, exosomes and other EVs are found in biological fluids including saliva, blood, urine and cerebrospinal fluid. Exosomes are similar to liposomes in terms of consisting of bilayered phospholipids, but the biogenesis of exosomes ensures their biocompatibility and low toxicity. It also significantly complicates pharmaceutical development, production and safety profiling=-oiuy678lkbvc(immunogenicity, and potential biological impurities).

[0040] As used herein, the terms "treatment," "treating," and the like, refer to obtaining a desired pharmacologic and / or physiologic effect. The effect may be prophylactic in terms of completely or partially preventing a disease or symptom thereof and / or may be therapeutic in terms of a partial or complete cure for a disease and / or adverse affect attributable to the disease. " Treatment," as used herein, covers any treatment of a disease in a mammal, particularly in a human, and includes: (a) preventing the disease from occurring in a subject which may be predisposed to the disease or at risk of acquiring the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; and (c) relieving the disease, i.e., causing regression of the disease.

[0041] Other technical terms used herein have their ordinary meaning in the art that they are used, as exemplified by a variety of technical dictionaries. The particular values andPATENT EX1-005WOconfigurations discussed in these non-limiting examples can be varied and are cited merely to illustrate at least one embodiment and are not intended to limit the scope thereof.DETAILED DESCRIPTION

[0042] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the subject technology as claimed. Additional features and advantages of the subject technology are set forth in the description below, and in part will be apparent from the description, or may be learned by practice of the subject technology. The advantages of the subject technology will be realized and attained by the structure particularly pointed out in the written description and claims hereof.

[0043] Intradermal drug delivery has several advantages to oral administration. For example, intradermal injection can improve the bioavailability of drugs. Side-effects that result from oral administration are generally absent or reduced. Embodiments include devices and methods that use microneedles for intracutaneous drug delivery.

[0044] Microneedles are micron-sized needles (e.g., from about 25 to 1500 pm in height) made of a variety of materials and shapes. They can be applied to the skin to create micron-sized transport pathways to enhance delivery of a drug molecule. The disclosure relates to adhesive patch assemblies that include an adhesive patch with an array of microneedles, and more particularly, to assemblies that include an active agent coated microneedle array that can be used to deliver the active agent through the skin.Microneedle Array Patch (MAP)

[0045] In aspects, the MAPs described herein include microneedles that penetrate the skin of a subject. In aspects, at least a portion of the MAP (e.g., the perimeter portion) is an adhesive patch material. The adhesive patch material can include an adhesive or tacky substance to adhere to skin.

[0046] FIG. 1 A depicts a microneedle array patch (MAP) according to aspects. It includes a microneedle array mounted on the center of an adhesive patch (e.g., 26.5PATENT EX1-005WOmm diameter). The adhesive patch can include backing film and medical adhesive tape. The MAP can be positioned in an applicator insert during storage and adheres to the skin following application. The MAPs described herein can be applied directly to the skin with manual pressure. Alternatively, they can be applied using a device as described herein.

[0047] FIG. 1 B depicts measurements of individual microneedles. In aspects, each of the microneedles is about 750 microns in length. In aspects, microneedles are separated from each other by a distance of about 650 microns. However, one skilled in the art will recognize that the size / distances can be adjusted based on the intended use.

[0048] The MAPs described herein can be used to administer large and small molecules. The transfer of active agents into the skin and into capillaries or lymphatic vessels is rapid and consistent. Shallow penetration of the microneedles minimizes stimulation of nerve endings. The MAPs are convenient, discreet and easy to use.

[0049] Applicant has optimized parameters that impact the delivered dose of an agent. In aspects, up to 1 mg of delivered content is delivered with high efficiency. This is significantly more than the expected dose of a vaccine (e.g., SARS-CoV-2 circRNA vaccine) that can be administered to a patient. For comparison, traditional intramuscular delivery of mRNA vaccines via needle injection, utilizes between 10 and 30μg of mRNA.Solid Microneedles

[0050] In aspects, the MAPs use solid microneedles. The solid microneedles can be implemented using a two-step application process. The needles puncture the skin and are then removed so the punctures can be covered with a transdermal patch. These tiny punctures allow the drug solution to seep deeper into the skin. Solid microneedles are often made of polymer, silicon or stainless steel.Coated Microneedles

[0051] In aspects, the MAPs use coated microneedles. Coated microneedles can be embedded in the skin for extended periods (e.g., hours) so the outer coating dissolvesPATENT EX1-005WOinto the epidermis or dermis. More typically, the coated formulation dissolves within the skin in a few minutes. After the formulation has been dissolved in the skin (i.e., the epidermis or dermis), the microneedle patch can be removed and disposed.

[0052] A benefit of coated microneedles is that they reduce the application into a single step. However, conventional microneedles are limited in how much solution they can deliver based on the thin coating and the amount retained during the initial puncture.Hollow Microneedles

[0053] In aspects, the MAPs use hollow microneedles. Hollow microneedles can puncture the skin and deliver liquid drug formulations from a drug reservoir. Hollow microneedles facilitate the flow of the liquid drug solution down through the needle and into the skin. Like coated microneedles, they simplify the application into a simple, one-step process but are eventually removed from the body.

[0054] Hollow microneedles can be prone to leakage and clogging. They also do not have the same stability advantages as solid coated MAPs. Yet, they are highly applicable to disease diagnosis practices and can deliver large doses of drug solutions.Dissolvable Microneedles

[0055] In aspects, the MAPs use dissolvable microneedles. Dissolvable microneedles gradually dissolve after they are lodged into the skin. Dissolvable microneedles are typically made of sugar and hydrophilic polymers. These are also applied using an adhesive patch, but the needles dissolve in the skin during the patch wear period.

[0056] Dissolvable microneedles can be made from a water-soluble polymer. Watersoluble polymers include, for example, hydroxyethyl starch, dextran, chondroitin sulfate, sodium chondroitin sulfate, sodium hyaluronate, carboxymethyl cellulose, polyvinylpyrrolidone, polyoxyethylene polyoxypropylene glycol, polyethylene glycol, and polyvinyl alcohol).

[0057] Dissolving microneedles can be more expensive and difficult to manufacture. Further, it may be necessary for patient to wear a MAP for an extended period of time (e.g., hours or days) as the needles slowly dissolve..PATENT EX1-005WOHydrogel-forming Microneedles

[0058] In aspects, the MAPs use hydrogel-forming microneedles. They can penetrate the skin and swell when they encounter the body's natural fluids. As they swell, they expand and release agents.

[0059] Unlike the dissolvable variety, hydrogel-forming microneedles are removed along with the adhesive patch once the drug solution has been distributed. Conventionally, they have been used to deliver hydrophilic drugs like caffeine and high-dose drugs like ibuprofen.Patch Applicator and Kinetic Applicator

[0060] FIG. 2A depicts a top view of a microneedle array patch (MAP) 105 and applicator insert 110, according to embodiments. Similarly, FIG. 2B depicts a bottom view of a microneedle array patch (MAP) and applicator insert.

[0061] The patch applicator ensures a high degree of uniformity in the patch application process. Aspects include a reusable applicator to achieve reliable adhesion and dosing. It can be used by a healthcare provider as well as a patient at home. The patch is effective at ensuring the required pressure for patch adhesion, across different skin types that can vary by thickness, age, moisture content, and placement.

[0062] FIG. 3 depicts a microneedle array patch (MAP) kinetic applicator, according to embodiments. The MAPs described herein can be applied directly to the skin with manual pressure (i.e., without using a device). However, use of an application device can provide advantages. An application device can ensure adequate and consistent insertion force and velocity. It can also ensure that a MAP is applied at a proper angle and improve general usability. The device can be used repeatedly by a patient or healthcare professional.

[0063] In aspects, a MAP is applied to a patient by the following steps:a) prime applicatorb) open patch / array assemblyc) insert patch / array into applicatorPATENT EX1-005WOd) place device on skine) activate (automatically applies patch / array to skin at correct force / velocity) f) store applicator for next use

[0064] Alternatively, a MAP can be applied using a low-cost disposable applicator with integrated coated patch assembly. It can be applied via the following steps:a) remove from packaging (patch / array assembly is built into applicator avoiding any assembly for the end user)b) place on skinc) push to activate (automatically applies patch / array to skin at correct force / velocity)d) Dispose

[0065] In aspects, the adhesive used in the patch has sufficient tack to maintain adhesion to the skin throughout the wear period (e.g., five to ten minutes). The MAP is designed to be flexible near the edges, to allow release from the tabs in the insert and to conform to the contours of the skin during wear.Circular RNAs

[0066] Circular RNAs (circRNAs) are a class of single-stranded RNAs with covalently linked head-to-tail topology. Unlike the linear mRNA vaccine which applications were limited by its instability, inefficiency, and innate immunogenicity, circRNA vaccine which incorporate internal ribosome entry sites (IRESs) and open reading frame (ORF) provides an improved approach to RNA-based vaccination with safety, stability, simplicity of manufacture, and scalability.

[0067] In embodiments the MAPs described herein are used to administer a circRNA vaccine. In aspects, the vaccine elicits an immune response against a bacteria.Examples of bacteria include, for example, Neisseria meningitidis, Streptococcus pneumoniae, Streptococcus pyogenes, Moraxella catarrhalis, Bordetella pertussis, Staphylococcus aureus, Clostridium tetani, Corynebacterium diphtheriae, Pseudomonas aeruginosa, Streptococcus agalactiae, Chlamydia trachomatis, Chlamydia pneumoniae, Helicobacter pylori, Escherichia coli, Bacillus anthracis, Yersinia pestis, StaphylococcusPATENT EX1-005WOepidermis, Clostridium perfringens or Clostridium botulinums, Legionella pneumophila, Coxiella burnetii, Brucella, (e.g., B. abortus, B.canis, B.melitensis, B.neotomae, B.ovis, B.suis, B.pinnipediae), Francisella, (e.g., F.novicida, F.philomiragia, F.tularensis).Neisseria gonorrhoeae, Treponema pallidum, Haemophilus ducreyi, Enterococcus faecalis or Enterococcus faeciumi, Staphylococcus saprophyticus, Yersinia enterocolitica, Mycobacterium tuberculosis, Rickettsia, Listeria monocytogenes, Vibrio cholerae, Salmonella typhi, Borrelia burgdorferi, Porphyromonas gingivalis and Klebsiella.

[0068] In aspects, the vaccine elicits an immune response against a virus. Examples of viruses include, for example, Orthomyxovirus, Paramyxoviridae viruses, Poxviridae, Picornavirus, enterovirus (e.g., EV71, coxsackie A or B), Bunyavirus, Orthobunyavirus (e.g., California encephalitis virus, a Phlebovirus, such as Rift Valley Fever virus, or a Nairovirus, such as Crimean-Congo hemorrhagic fever virus), Heparnavirus (e.g., hepatitis A virus), Filovirus, Togavirus (e.g., Rubivirus, an Alphavirus, or an Arterivirus. rubella virus, Flavivirus (e.g., Tick-borne encephalitis (TBE) virus, Dengue ( types 1, 2, 3 or 4) virus, Yellow Fever virus, Japanese encephalitis virus, Kyasanur Forest Virus, West Nile, Encephalitis virus (e.g, St. Louis encephalitis virus, Russian spring-summer encephalitis virus, Powassan encephalitis virus), Pestivirus (e.g., Bovine viral diarrhea, “BVDV” Classical swine fever “CSFV” or Border disease), Hepadnavirus (e.g., Hepatitis B virus, hepatitis C virus, delta hepatitis virus hepatitis E virus, or hepatitis G virus, Rhabdovirus (e.g. a Rabies virus and Vesiculovirus), Caliciviridae (e.g., Norovirus and Norwalk-like Viruses, such as Hawaii Virus and Snow Mountain Virus), Coronavirus: (e.g., SARS coronavirus, avian infectious bronchitis (IBV), Mouse hepatitis virus (MEV), and Porcine transmissible gastroenteritis virus (TGEV), Retrovirus (e.g., Oncovirus, Lentivirus (e.g. HIV-1 or HIV-2) or a Spumavirus, Reovirus: (e.g., Orthoreovirus, a Rotavirus, an Orbivirus, or a Coltivirus) Parvovirus, Herpesvirus: (e.g., Herpes Simplex Viruses (HSV) (e.g. HSV types 1 and 2), Varicella-zoster Virus (VZV), Epstein-Barr virus (EBV, Cytomegalovirus (CMV), Human Herpesvirus 6 (HHV6), Human Herpesvirus 7 (HHV7), and Human Herpesvirus 8 (HHV8), Papovaviruses and Adenovirus.

[0069] In aspects, the vaccine elicits an immune response against a fungus. FungalPATENT EX1-005WOimmunogens can be derived from Dermatophytres, including: Epidermophyton floccusum, Microsporum audouini, Microsporum canis, Microsporum distortum, Microsporum equinum, Microsporum gypsum, Microsporum nanum, Trichophyton concentricum, Trichophyton equinum, Trichophyton gallinae, Trichophyton gypseum, Trichophyton megnini, Trichophyton mentagrophytes, Trichophyton quinckeanum, Trichophyton rubrum, Trichophyton schoenleini, Trichophyton tonsurans, Trichophyton verrucosum, T. verrucosum var. album, var. discoides, var. ochraceum, Trichophyton violaceum, and / or Trichophyton favifornie; or from Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Aspergillus nidulans, Aspergillus terreus, Aspergillus sydowi, Aspergillus flavatus, Aspergillus glaucus, Blastoschizomyces capitatus, Candida albicans, Candida enolase, Candida tropicalis, Candida glabrata, Candida krusei, Candida parapsilosis, Candida stellatoidea, Candida kusei, Candida parakwsei, Candida lusitaniae, Candida pseudotropicalis, Candida guilliermondi, Cladosporium carrionii, Coccidioides immitis, Blastontyces dermatidis, Cryptococcus neoformans, Geotrichum clavatum, Histoplasma capsulatum, Klebsiella pneumoniae, Microsporidia, Encephalitozoon spp., Septata intestinalis and Enterocytozoon bieneusi; the less common are Brachiola spp, Microsporidium spp., Nosema spp., Pleistophora spp., Trachipleistophora spp., Vittaforma spp Paracoccidioides brasiliensis, Pneumocystis carinii, Pythiumn insidiosum, Pityrosporum ovale, Sacharomyces cerevisae, Saccharomyces boulardii, Saccharomyces pombe, Scedosporium apiosperum, Sporothrix schenckii, Trichosporon beigelii, Toxoplasma gondii, Penicillium marneffei, Malassezia spp., Fonsecaea spp., Wangiella spp., Sporothrix spp., Basidiobolus spp., Conidiobolus spp., Rhizopus spp., Mucor spp, Absidia spp, Mortierella spp, Cunninghamella spp, Saksenaea spp., Alternaria spp, Curvularia spp, Helminthosporium spp, Fusarium spp, Aspergillus spp, Penicillium spp, Rhizoctonia spp, Paecilomyces spp, Pithomyces spp. and Cladosporium ssp.

[0070] In aspects, the vaccine elicits a response against a parasite. The parasite can be from, for example, the Plasmodium genus, such as P. falciparum, P.vivax, P.malariae or P. ovale. Thus, the invention can be used for immunizing against malaria. In aspects, the immunogen elicits an immune response against a parasite from the Caligidae family (e.g., from the Lepeophtheirus and Caligus genera (sea lice) such as LepeophtheirusPATENT EX1-005WOsalmonis or Caligus rogercresseyi).

[0071] In embodiments, the MAPs described herein are used to administer a circRNA vaccine against cancer. The immunogen can be a tumor antigen selected from: (a) cancer-testis antigens such as NY-ESO-1, SSX2, SCP1 as well as RAGE, BAGE, GAGE and MAGE family polypeptides, for example, GAGE-1, GAGE-2, MAGE-1, MAGE-2, MAGE-3, MAGE-4, MAGE-5, MAGE-6, and MAGE-12 (which can be used, for example, to address melanoma, lung, head and neck, NSCLC, breast, gastrointestinal, and bladder tumors; (b) mutated antigens, for example, p53 (associated with various solid tumors, e.g., colorectal, lung, head and neck cancer), p21 / Ras (associated with, e.g., melanoma, pancreatic cancer and colorectal cancer), CDK4 (associated with, e.g., melanoma), MUM1 (associated with, e.g., melanoma), caspase-8 (associated with, e.g., head and neck cancer), CIA 0205 (associated with, e.g., bladder cancer), HLA-A2-R1701, beta catenin (associated with, e.g., melanoma), TCR (associated with, e.g., T-cell non-Hodgkins lymphoma), BCR-abl (associated with, e.g., chronic myelogenous leukemia), triosephosphate isomerase, KIA 0205, CDC-27, and LDLR-FUT; (c) overexpressed antigens, for example, Galectin 4 (associated with, e.g., colorectal cancer), Galectin 9 (associated with, e.g., Hodgkin's disease), proteinase 3 (associated with, e.g., chronic myelogenous leukemia), WT 1 (associated with, e.g., various leukemias), carbonic anhydrase (associated with, e.g., renal cancer), aldolase A (associated with, e.g., lung cancer), PRAME (associated with, e.g., melanoma), HER-2 / neu (associated with, e.g., breast, colon, lung and ovarian cancer), mammaglobin, alpha-fetoprotein (associated with, e.g., hepatoma), KSA (associated with, e.g., colorectal cancer), gastrin (associated with, e.g., pancreatic and gastric cancer), telomerase catalytic protein, MUC-1 (associated with, e.g., breast and ovarian cancer), G-250 (associated with, e.g., renal cell carcinoma), p53 (associated with, e.g., breast, colon cancer), and carcinoembryonic antigen (associated with, e.g., breast cancer, lung cancer, and cancers of the gastrointestinal tract such as colorectal cancer); (d) shared antigens, for example, melanoma-melanocyte differentiation antigens such as MART-1 / Melan A, gp100, MCIR, melanocyte-stimulating hormone receptor, tyrosinase, tyrosinase related protein-1 / TRPI and tyrosinase related protein-2 / TRP2 (associated with, e.g., melanoma); (e) prostate associated antigens such as PAP, PSA, PSMA, PSH-P1, PSM-PATENT EX1-005WOP1, PSM-P2, associated with e.g., prostate cancer; (f) immunoglobulin idiotypes (associated with myeloma and B cell lymphomas, for example). In certain embodiments, tumor immunogens include, but are not limited to, p15, Hom / Mel-40, H-Ras, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, Epstein Barr virus antigens, EBNA, human papillomavirus (HPV) antigens, including E6 and E7, hepatitis B and C virus antigens, human T-cell lymphotropic virus antigens, TSP-180, p185erbB2, p180erbB-3, c-met, mn-23H1, TAG-72-4, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, p16, TAGE, PSCA, CT7, 43-9F, 5T4, 791 Tgp72, beta-HCG, BCA225, BTAA, CA 125, CA 15-3 (CA 27.29\BCAA), CA 195, CA 242, CA-50, CAM43, CD68\KP1, CO-029, FGF-5, Ga733 (EpCAM), HTgp-175, M344, MA-50, MG7-Ag, M0V18, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, TA-90 (Mac-2 binding protein / cyclophilin C-associated protein), TAAL6, TAG-72, TLP, TPS, and the like.Dermal Expression of Proteins Using Circular RNA

[0072] The dermis into which the needles of the microarray penetrate consists of multiple cells types, including leukocytes, lymphoid cells and myeloid cells. The cell types include neutrophils, granulocytes, macrophages, Kupffer cells, innate lymphoid cells, natural killer cells, dendritic cells, B cells and T cells.

[0073] In an embodiment, a microneedle patch comprises a nanoparticle comprising a circular RNA encoding one, two, three, four, five, six, seven, eight, nine, ten or more different proteins and / or peptides. In a further embodiment a nanoparticle is a lipid nanoparticle, a dendrimer, a polymer nanoparticle and / or a solid support structure that a circular RNA. The solid support structure can be a solid material, including gold, silver, titanium, a plastic, silicon, platinum or other solid medium. The solid support structure can be in the shape of a ball, a square, a rectangle, an ellipse, an amoeba or other shape. The surface of the solid support can be smooth, rough or both rough and smooth. In an embodiment, the solid support shields the circular RNA, whether encapsulated within a nanoparticle or not from degradation.

[0074] The cells that are able to take in the nanoparticle comprising a circular RNA following application of a microneedle, including a microneedle containing patch, to aPATENT EX1-005WOdermis of a patient receiving the microneedle include: activated myeloid cells, adipocyte-like stromal cells, capillary endothelium, fibroblasts, lymphatic-associated macrophages, oxidative muscle fibers, B cells, T cells, dendritic cells, neutrophils, resident macrophages and Schwann cells.

[0075] In an embodiment a nanoparticle comprising a circular RNA that encodes one or more proteins and / or peptides is administered to a patient by application of a microneedle to the patient’s dermis. In a further embodiment, the protein and / or peptide encoded by the circular RNA is taken up by activated myeloid cells, adipocyte-like stromal cells, capillary endothelium, fibroblasts, lymphatic-associated macrophages, oxidative muscle fibers, B cells, T cells, macrophages, dendritic cells, natural killer cells, neutrophils, resident macrophages and Schwann cells. In an embodiment, the protein and / or peptide is a vaccine, a chimeric antigen receptor (CAR), an antibody, an antibody fragment (e.g. sfv, VHH, VH), a therapeutic, a vitamin, an antibody, a contractile protein, an enzyme, a hormonal protein, a storage protein, a structural protein, a protective protein (e.g. an immune response protein, including a cytokine), a structural protein and / or a transport protein.

[0076] In an embodiment, a microneedle patch comprising one or more microneedles is applied to the skin of a patient, wherein the microneedles are coated with a formulation comprising a nanoparticle comprising one or more circular RNA’s comprising one or more proteins and / or peptides and the nanoparticles comprising the one or more circular RNAs are taken up and internalized by one or more of an activated myeloid cell, an adipocyte-like stromal cell, a capillary endothelium, a fibroblast, a lymphatic-associated macrophage, an oxidative muscle fiber, a cell, a T cell, a dendritic cell, a neutrophil, a resident macrophage and a Schwann cell, wherein, the cells that took up the one or more circular RNAs express the one or more proteins or peptides encoded by the one or more circular RNAs. In an embodiment, the T cells is a helper T cell, a regulatory T cell, a killer T cell, a natural killer T cell, a CD4+cell and / or a CD8+cell. In an embodiment, a protein that is expressed by a cell in the dermis is capable of treating a cancer, a fibrosis, an autoimmune diseases, an infectious disease, a cardiovascular disease.PATENT EX1-005WO

[0077] In a further embodiment, a cancer is treated by expressing a protein and / or a peptide encoded by a circular RNA that is administered by a microneedle to a cell residing in the dermis of a patient. In another embodiment, a cancer is treated by administering a nanoparticle comprising a circular RNA to the dermis of a patient and wherein, the nanoparticle comprising a circular RNA is transported (e.g. through the lymphatic system, a blood vessel, movement of interstitial fluid) to a distal site not that of the site of administration by the microneedle. In an embodiment, the distal site is an organ of a patient. The organ includes one or more of the liver, a kidney, the heart, a lung, the brain, a bone marrow, a thymus, the pancreas, the spleen, a lymph node, a Peyer’s Patch, the stomach, the colon, the intestine (large and / or small), the vagina, the testes, an ovary, a pituitary gland, a saliva gland, a muscle, a skin, a nasal tissue and the prostate.

[0078] In a further embodiment, an autoimmune disease is treated by expressing a protein and / or a peptide encoded by a circular RNA that is administered by a microneedle to a cell residing in the dermis of a patient. In another embodiment, an autoimmune disease is treated by administering a nanoparticle comprising a circular RNA to the dermis of a patient and wherein, the nanoparticle comprising a circular RNA is transported (e.g. through the lymphatic system, a blood vessel, movement of interstitial fluid) to a distal site not that of the site of administration by the microneedle. In an embodiment, the site is an organ of a patient. The organ includes one or more of the liver, a kidney, the heart, a lung, the brain, a bone marrow, a thymus, the pancreas, the spleen, a lymph node, a Peyer’s Patch, the stomach, the colon, the intestine (large and / or small), the vagina, the testes, an ovary, a pituitary gland, a saliva gland, a muscle, a skin, a nasal tissue and the prostate.

[0079] In a further embodiment, an infectious disease is treated by expressing a protein and / or a peptide encoded by a circular RNA that is administered by a microneedle to a cell residing in the dermis of a patient. In another embodiment, an infectious disease is treated by administering a nanoparticle comprising a circular RNA to the dermis of a patient and wherein, the nanoparticle comprising a circular RNA is transported (e.g. through the lymphatic system, a blood vessel, movement of interstitial fluid) to a distalPATENT EX1-005WOsite not that of the site of administration by the microneedle. In an embodiment, the site is an organ of a patient. The organ includes one or more of the liver, a kidney, the heart, a lung, the brain, a bone marrow, a thymus, the pancreas, the spleen, a lymph node, a Peyer’s Patch, the stomach, the colon, the intestine (large and / or small), the vagina, the testes, an ovary, a pituitary gland, a saliva gland, a muscle, a skin, a nasal tissue and the prostate.

[0080] In a further embodiment, a fibrosis disease is treated by expressing a protein and / or a peptide encoded by a circular RNA that is administered by a microneedle to a cell residing in the dermis of a patient. In another embodiment, a fibrosis disease is treated by administering a nanoparticle comprising a circular RNA to the dermis of a patient and wherein, the nanoparticle comprising a circular RNA is transported (e.g. through the lymphatic system, a blood vessel, movement of interstitial fluid) to a distal site not that of the site of administration by the microneedle. In an embodiment, the site is an organ of a patient. The organ includes one or more of the liver, a kidney, the heart, a lung, the brain, a bone marrow, a thymus, the pancreas, the spleen, a lymph node, a Peyer’s Patch, the stomach, the colon, the intestine (large and / or small), the vagina, the testes, an ovary, a pituitary gland, a saliva gland, a muscle, a skin, a nasal tissue and the prostate.

[0081] In a further embodiment, a cardiovascular disease is treated by expressing a protein and / or a peptide encoded by a circular RNA that is administered by a microneedle to a cell residing in the dermis of a patient. In another embodiment, a cardiovascular disease is treated by administering a nanoparticle comprising a circular RNA to the dermis of a patient and wherein, the nanoparticle comprising a circular RNA is transported (e.g. through the lymphatic system, a blood vessel, movement of interstitial fluid) to a distal site not that of the site of administration by the microneedle. In an embodiment, the distal site is an organ of a patient. The organ includes one or more of the heart and / or a blood vessel.

[0082] In an embodiment, a microneedle is coated with a formulation comprising a circular RNA. In a further embodiment, the formulation comprising a circular RNA is aPATENT EX1-005WOliquid. In another embodiment, the formulation that was a liquid becomes a solid or semi-solid on the microneedle. In an embodiment, the formulation comprising a circular RNA forms a droplet on the microneedle. In an embodiment, the circular RNA is part of a nanoparticle. In a further embodiment, the circular RNA is inside a nanoparticle. In an embodiment, the circular RNA is encapsulated by a nanoparticle. In an embodiment, a circular RNA that is inside and / or encapsulated by a nanoparticle is located in a formulation on the outer surface of a microneedle.

[0083] In an embodiment, a microneedle comprising one or more circular RNA(s) (whether administered inside or encapsulated by a nanoparticle or not) is / are administered to a patient. In an embodiment, the microneedle comprising one or more circular RNA(s) is / are administered to a skin of a patient and the one or more circular RNA(s) is / are administered to the dermis. In a further embodiment, the one or more circular RNA(s) that has / have been applied to the dermis is / are taken up by a cell. In a further embodiment, the cell expresses the one or more proteins and / or peptides encoded in the one or more circular RNA(s).In an embodiment, the cell that has taken up the circular RNA(s) remain / remains in the dermis. In another embodiment, the cell that has taken up the one or more circular RNA(s) migrates out of the dermis to another location in the patient, including an organ, the circulatory system, and / or the lymphatic system.

[0084] In an embodiment, a cell in the dermis that has taken up / in a circular RNA produces / expresses a protein and / or a peptide encoded by the one or more circular RNA(s). In a further embodiment, the production / expression occurs shortly after administration of a circular RNA into the dermis by a microneedle. In a further embodiment, expression / production starts 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 60 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 1 day or more after administration.PATENT EX1-005WO

[0085] In an embodiment a cell in the dermis that takes up / in a circular RNA administered by a microneedle patch produces / expresses a protein and / or a peptide for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 30 days, 35 days, 40 days, 45 days, 50 days, 55 days, 60 days, 65 days, 70 days, 75 days or more after administration.

[0086] In an embodiment a cell in the dermis that takes up / in a circular RNA administered by a microneedle patch produces / expresses a protein and / or a peptide for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, at least 22 days, at least 23 days, at least 24 days, at least 25 days, at least 30 days, at least 35 days, at least 40 days, at least 45 days, at least 50 days, at least 55 days, at least 60 days, at least 65 days, at least 70 days, at least 75 days or more after administration.

[0087] In an embodiment a cell in the dermis that takes up / in a circular RNA administered by a microneedle patch produces / expresses a protein and / or a peptide for no more than 1 day, no more than 2 days, no more than 3 days, no more than 4 days, no more than 5 days, no more than 6 days, no more than 7 days, no more than 8 days, no more than 9 days, no more than 10 days, no more than 11 days, no more than 12 days, no more than 13 days, no more than 14 days, no more than 15 days, no more than 16 days, no more than 17 days, no more than 18 days, no more than 19 days, no more than 20 days, no more than 21 days, no more than 22 days, no more than 23 days, no more than 24 days, no more than 25 days, no more than 30 days, no more than 35 days, no more than 40 days, no more than 45 days, no more than 50 days, no more than 55 days, no more than 60 days, no more than 65 days, no more than 60 days, no more than 65 days, no more than 70 days, no more than 75 days or more after administration.PATENT EX1-005WO

[0088] In an embodiment a cell in the dermis that takes up / in a circular RNA administered by a microneedle patch produces / expresses a protein and / or a peptide for about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 1 days, about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, about 21 days, about 22 days, about 23 days, about 24 days, about 25 days, about 30 days, about 35 days, about 40 days, about 45 days, about 50 days, about 55 days, about 60 days, about 65 days, about 60 days, about 65 days, about 70 days, about 75 days or more after administration.EXAMPLES

[0089] The following non-limiting examples are provided for illustrative purposes only in order to facilitate a more complete understanding of representative embodiments now contemplated. These examples are intended to be a mere subset of all possible contexts in which the components of the formulation may be combined. Thus, these examples should not be construed to limit any of the embodiments described in the present specification, including those pertaining to the type and amounts of components of the formulation and / or methods and uses thereof.Example 1Effectiveness of Delivering circRNA Vaccine using MAPs

[0090] In this study, Applicant examined the effect of various excipients on the intradermal delivery and expression of a circular RNA vaccine using solid microneedle array patches. Five different formulations (Formulations F, G, H, J and K) of one circular RNA vaccine were administered to ten pigs using solid microneedle array patches. Serum and tissue samples were harvested from the pigs at the completion of the study and sent to analyzed. All animal testing was conducted in accordance with an IACUC approved protocol.

[0091] The solid microneedle array (depicted FIG. 1) was injection molded with USP Class VI rated liquid crystal polymer (LCP) resin (Ticona, Vectra® MT1300). The microneedle array used in this study was a 1-mm thick circular disc with an overallPATENT EX1-005WOsurface area of 1.27 cm2or -12.7 mm in diameter, containing approximately 216 pyramid-shaped, 750-micron long microstructures. As shown, the microneedles are spaced approximately 650 microns apart (tip to tip) in a geometric pattern.MAP Coating

[0092] Five lots of coated patches were manufactured for application to the swine using a stock circRNA / LNP vaccine solution. The physical properties and compositional elements of the stock circRNA / LNP vaccine solution are shown below (Table 1).Generally, the formulation and coating of the microneedle array patches with circRNA / LNP was successful. Specific formulation and coating information for each lot can be found in the respective sections below.Table 1: Stock circRNA / LNP Vaccine SolutionPhysical Property / Composition ValuecircRNA Concentration (mg / mL) 1~LNP:mRNA Ratio (w:w) 19Sucrose Content (%w / w) 10~PBS (%w / w) 0.98-Density (g / mL) 1MAP Formulation F - Hydroxyethylcellulose

[0093] The composition of Formulation F is shown below (Table 2). The stock circRNA / LNP vaccine solution was removed from -80°C and thawed at room temperature (~20°C) for approximately 45 minutes prior to formulation. 500 pL of stock circRNA / LNP solution was added to a 3 mL syringe. HEC was then added as dried powder directly to the vaccine solution (HEC 250 M followed by HEC 250 L) and lightly shaken by hand to disperse the powders (total formulation time -25-30 minutes). The formulation syringe was placed on a roller to gently mix (15 RPM) for -3 hours at room temperature (~20°C) before being moved to 5°C and gently mixed (12 RPM) for another -7.5 hours. The formulation was then transferred to the isolator and used to coat the microneedle array patches. Coated array patches were sealed in a PETG pod with a Tyvek lid and further sealed in a foil pouch with a desiccant (total coating and packaging time -60-90 minutes). No issues were encountered during mixing and the resultingPATENT EX1-005WOformulation was a homogeneous hazy suspension. Coating of the microneedle array patches was completed with no issues and resulted in uniform coatings across the arrays.Table 2: Formulation F CompositionIngredient ~Coating Formulation ~Dried Coating Formulation Composition (%) Composition (%)circRNA 0.09 0.532LNP 1.80 10.106Sucrose 9.45 53.191PBS 0.93 5.213HEC 250 M (MW: ~720K) 0.50 2.814HEC 250 L (MW: ~90K) 5.00 28.143Sterile Water 82.23 N / AMAP Formulation G - HydroxypropylcelluloseThe composition of Formulation G is shown below (Table 3). The stock circRNA / LNP vaccine solution was removed from -80 °C and thawed at room temperature (~20 °C) for approximately 1.25 hours prior to formulation. 500 pL of stock circRNA / LNP solution was added to a 3 mL syringe. HPC was then added as dried powder directly to the vaccine solution (HPC MF followed by HPC ELF) and lightly shaken by hand to disperse the powders (total formulation time ~25-30 minutes). The formulation syringe was placed on a roller to gently mix (15 RPM) for ~3 hours at room temperature (~20°C) before being moved to 5°C and gently mixed (12 RPM) for another ~23 hours. The formulation was then transferred to the isolator and used to coat the microneedle array patches. Coated array patches were sealed in a PETG pod with a Tyvek lid and further sealed in a foil pouch with a desiccant (total coating and packaging time ~60-90 minutes). No significant issues were encountered during mixing and the resulting formulation was a generally homogeneous hazy suspension. Coating of the microneedle array patches was completed with no issues and uniform coatings across the array were achieved.Table 3: Formulation G CompositionIngredient ~Coating Formulation ~Dried Coating Formulation Composition (%) Composition (%)circRNA 0.09 0.532LNP 1.80 10.106PATENT EX1-005WOSucrose 9.45 53.191PBS 0.93 5.213HPC MF (MW: ~850K) 0.50 2.814HPC ELF (MW: ~40K) 5.00 28.143Sterile Water 82.23 N / AMAP Formulation H - Polyvinyl Alcohol

[0094] The composition of Formulation H is shown below (Table 4). The stock circRNA / LNP vaccine solution was removed from -80 °C and thawed at room temperature (~20 °C) for approximately 45 minutes prior to formulation. 500 pL of stock circRNA / LNP solution was added to a 3 mL syringe. Approximately 150 pL of a concentrated (22% w / w) PVA solution was then added to the vaccine solution and lightly shaken by hand to disperse the solutions (total formulation time ~15 minutes). The formulation syringe was placed on a roller to gently mix (15 RPM) for ~3 hours at room temperature (~20°C) before being moved to 5°C and gently mixed (12 RPM) for another ~8 hours. The formulation was then transferred to the isolator and used to coat the microneedle array patches. Coated array patches were sealed in a PETG pod with a Tyvek lid and further sealed in a foil pouch with a desiccant (total coating and packaging time ~60-90 minutes). No significant issues were encountered during mixing and the resulting formulation was a homogeneous hazy suspension. Coating of the microneedle array patches was completed with no issues. Uniform coatings across the arrays were achieved. (FIG. 4).Table 4: Formulation H CompositionIngredient ~Coating Formulation ~Dried Coating Formulation Composition (%) Composition (%)circRNA 0.08 0.494LNP 1.43 9.387Sucrose 7.52 49.407PBS 0.74 4.842PVA, 88% Hydrolyzed (MW: 145-180K) 5.46 35.870Sterile Water 84.78 N / AMAP Formulation J - Polyvinylpyrrolidone

[0095] The composition of Formulation J is shown below (Table 5). The stockPATENT EX1-005WOcircRNA / LNP vaccine solution was removed from -80 °C and thawed at room temperature (~20 °C) for approximately 1 hour prior to formulation. 500 pL of stock circRNA / LNP solution was added to a 3 mL syringe. Approximately 140 pL of a concentrated (25% w / w) PVP PBS solution was then added to the vaccine solution and lightly shaken by hand to disperse the solutions (total formulation time -15-20 minutes). The formulation syringe was placed on a roller to gently mix (15 RPM) for -2.5 hours at room temperature (~20°C) before being moved to 5°C and gently mixed (12 RPM) for another -6 hours. The formulation was then transferred to the isolator and used to coat the microneedle array patches. Coated array patches were sealed in a PETG pod with a Tyvek lid and further sealed in a foil pouch with a desiccant (total coating and packaging time -60-90 minutes). No significant issues were encountered during mixing and the resulting formulation was a homogeneous hazy suspension. Coating of the microneedle array patches was completed with no issues. Uniform coatings across the arrays were achieved.Table 5: Formulation J CompositionIngredient ~Coating Formulation ~Dried Coating Formulation Composition (%) Composition (%)circRNA 0.08 0.478LNP 1.45 9.091Sucrose 7.65 47.847PBS 0.92 5.742PVP K90 (MW: 900-1200K) 5.89 36.842Sterile Water 84.02 N / AMAP Formulation K- Polyvinyl Alcohol / Polyvinylpyrrolidone

[0096] The composition of Formulation K is shown below (Table 6). The stock circRNA / LNP vaccine solution was removed from -80 °C and thawed at room temperature (-20 °C) for approximately 45 minutes prior to formulation. 500 pL of stock circRNA / LNP solution was added to a 3 mL syringe. Approximately 78 pL of a concentrated (22% w / w) PVA solution was then added to the vaccine solution followed by approximately 68 pL of a concentrated (25% w / w) PVP PBS solution and lightly shaken by hand to disperse the solutions (total formulation time -15 minutes). The formulation syringe was placed on a roller to gently mix (15 RPM) for -2 hours at roomPATENT EX1-005WOtemperature (~20°C) before being moved to 5°C and gently mixed (12 RPM) for another -7 hours. The formulation was then transferred to the isolator and used to coat the microneedle array patches. Coated array patches were sealed in a PETG pod with a Tyvek lid and further sealed in a foil pouch with a desiccant (total coating and packaging time -60-90 minutes). No significant issues were encountered during mixing and the resulting formulation was a homogeneous hazy suspension. Coating of the microneedle array patches was completed with no issues. Uniform coatings across the arrays were achieved.Table 6: Formulation K CompositionIngredient “Coating Formulation ~Dried Coating Formulation Composition (%) Composition (%)circRNA 0.08 0.486LNP 1.44 9.225Sucrose 7.57 48.555PBS 0.82 5.244PVA, 88% Hydrolyzed (MW: 145-180K) 2.86 18.330PVP K90 (MW: 900-1200K) 2.83 18.160Sterile Water 84.41 N / AcircRNA MAP and Control MAP Application

[0097] Ten pigs received two vaccine-coated MAPs on the left outer thigh; and 2 blank MAPs on the right outer thigh All pigs were dosed on Day 0 and euthanized on Day 3 or Day 7 (i.e., 3 or 7 days after dosing). The study is summarized in Table 7.PATENT EX1-005WOTable 7: Study SummaryAnimal model Yorkshire pigAnimal weight 25 to 52 kg at dosingNumber of Animals 10RNA Patch Applications per animal 2 (two vaccine -coated MAPs on the left thigh)Blank MAP Applications per animal 2 (two blank MAPs on the right thigh)Vaccine Formulation Circular RNA vaccine-coated MAPNumber of Vaccine Formulations 5Blank non-coated MAPApplicator Reusable MAP applicator for 12.7mm-0 MAPSkin site(s) Outer ThighWear time 15 minutesImages of MAP sites post-removal skin site on DayOPre-study blood (each animal) 5 mL serum (1 mL aliquots)Post-study blood (each animal) 5 mL serum (1 mL aliquots)Necropsy Tissues and Organs (each vaccine site skin (frozen + NBF treated)animal) control site skin (frozen + NBF treated)liver (frozen + NBF treated)spleen (frozen + NBF treated)inguinal lymph node (frozen + NBF treated)popliteal fossa lymph node (frozen + NBF treated)thymus (frozen + NBF treated)MAP Application and Removal Procedure

[0098] Hair was removed from both outer thighs prior to the application of the MAP device to minimize complications during MAP insertion and adhesion. Skin test sites were selected based on location and absence of skin damage. The hair was first clipped using an electric clipper with a #40 blade and then wiped with gauze that had been sprayed with 70% IPA. The IPA solution was allowed to evaporate from the skin for at least 15 minutes prior to dosing.

[0099] A MAP was inserted into the applicator and the applicator was positioned over the skin application site per an application schedule. The operator gently radially stretched and tensioned the application site skin and then actuated the applicator to insert the MAP into the skin. While the MAP was inserted, 4 small tattoo marks were made with a hypodermic needle around the MAP(s) to indicate the boundaries of the microneedle application for necropsy at study termination.PATENT EX1-005WO

[0100] MAP units were removed after 15 minutes using a blunt forceps and carefully transferred to a tray to protect the microstructures. The operator swabbed each application site with a single damp polyester swab. After MAPs were removed from the left side, the animal was turned over and the process was repeated.Swabbing Post-Removal Application Sites

[0101] 0.20 mL RNase-free water containing RNasin (ribonuclease inhibitor; approximately 200U) was pipetted into each 20mL RNase-free container and cap the container. After removal of each MAP a polyester swab was grabbed with clean forceps, the swab container cap was removed and the swab was moistened with the diluent in the container. The skin zone was gently swabbed (two times up and down; turn swab over and two times right and left. Care was taken to not to leave drops on the skin. The swab was placed in the same 20mL RNase-free plastic container, the container was capped, and frozen by placing in contact with dry ice. The forceps was cleaned after each swabbing.Blood Collection, Serum Separation Method and Storage

[0102] Ten mL of whole blood was collected in a SST (serum separator tube), or similar from each pig. Blood was spun in a refrigerated centrifuge. Serum was collected, transferred in five 1 mL aliquots to a plastic cryovial and frozen on dry ice or in the ultralow temperature (-70 ° C) freezer. Dry ice or ultra-low temperature (-70°C) freezer.Necropsy – Application Site Skin and Control Site Skin

[0103] At necropsy, one full-thickness application site skin with underlying muscle (approximately 13mm X 13mm X at least 6mm muscle depth) and one control site skin with underlying muscle (approximately 13mm X 13mm X at least 6mm muscle depth) was excised, placed in separate labeled sealable RNase-free suitable containers, flash frozen with liquid nitrogen and stored at -70°C until shipment on dry ice for circular RNA and antigen analysis. Duplicate samples, one full-thickness application site skin with underlying muscle (approximately 13mm X 13mm X at least 6mm muscle depth) and one control site skin with underlying muscle (approximately 13mm X 13mm X at least 6mm muscle depth) was excised, placed in separate labeled sealable RNase-freePATENT EX1-005WOsuitable containers with 10% NBF and stored at 4°C for 48-72 hours followed by washing the tissue in 1x PBS (phosphate buffered saline) at room temperature 3X for 30 min each). PBS washed samples were stored at 4°C until shipment.Liver

[0104] At necropsy, 3 samples from each liver lobe were collected using a 12mm biopsy punch and placed in the same labelled RNase-free container (not more than 10mm thick), flash frozen with liquid nitrogen and stored at -70°C until shipment on dry ice for circular RNA and antigen analysis. Additionally, 3 samples from each liver lobe were collected using a 12mm biopsy punch and placed in the same labelled RNase-free container (not more than 10mm thick) with 10% NBF and stored at 4°C for 48-72 hours followed by washing the tissue in 1x PBS (phosphate buffered saline) at room temperature 3X for 30 min each). PBS washed samples were stored at 4°C until shipment.Spleen

[0105] At necropsy, the spleen was divided lengthwise to allow best visualization of the red and white pulp. Six (6) biopsy samples (12mm diameter X not thicker than 10mm to preserve the red and white pulp within a biopsy) were collected and placed in the same RNase-free container. Samples were flash frozen in liquid nitrogen and stored in ultralow freezer until shipment for RNA and / or antigenic expression testing.Additionally, 6 biopsy samples of the spleen (12mm diameter X not thicker than 10mm to preserve the red and white pulp within a biopsy) were collected and placed in the same RNase-free container with 10% NBF and stored at 4°C for 48-72 hours followed by washing the tissue in 1x PBS (phosphate buffered saline) at room temperature 3X for 30 min each). PBS washed samples were stored at 4°C until shipment.Inguinal Lymph Node

[0106] At necropsy, each lymph node was divided, right and left, into quarters. Two quarters (i.e., half of the node; each quarter should be no thicker than 10mm) from the same lymph node were placed together in the same RNase-free container. Samples were flash frozen in liquid nitrogen and stored in ultralow freezer until shipment for RNAPATENT EX1-005WOand / or antigenic expression testing. Additionally, the other two quarters (i.e., half of the node; each quarter should be no thicker than 10mm) from the same lymph node were placed together in the same RNase-free container with 10% NBF and stored at 4°C for 48-72 hours followed by washing the tissue in 1x PBS (phosphate buffered saline) at room temperature 3X for 30 min each). PBS washed samples were stored at 4°C until shipment.Popliteal Fossa Lymph Node

[0107] At necropsy, approximately one half of the popliteal fossa lymph node from each the left and right side was collected in separate labeled RNase-free containers. Samples were flash frozen in liquid nitrogen and stored in ultralow freezer until shipment for RNA and / or antigenic expression testing. Additionally, approximately one half of the popliteal fossa lymph node from each the left and right side was collected in separate labeled RNase-free containers with 10% NBF and stored at 4°C for 48-72 hours followed by washing the tissue in 1x PBS (phosphate buffered saline) at room temperature 3X for 30 min each). PBS washed samples were stored at 4°C until shipment.Thymus

[0108] At necropsy, approximately 1 cm3mid-level slice from the left and right thymus was collected. Three punch biopsies from both sides in the medulla were placed in RNAse-free containers labelled M4, M5, and M6. Three punch biopsies from both sides were place in RNAse-free containers labeled C4, C5, and C6. Samples were flash frozen in liquid nitrogen and stored in an ultralow freezer until shipment for RNA and / or antigenic expression testing. Additionally, three punch biopsies from both sides in the medulla were placed in RNAse-free containers labelled M1, M2, and M3. Three punch biopsies from both sides were place in RNAse-free containers labeled C1, C2, and C3 with 10% NBF and stored at 4°C for 48-72 hours followed by washing the tissue in 1x PBS (phosphate buffered saline) at room temperature 3X for 30 min each). PBS washed samples were stored at 4°C until shipment.

[0109] The samples were analyzed to confirm that each subject was effectivelyPATENT EX1-005WOimmunized. In conclusion, the study demonstrated:• a robust Immune response in all subjects• all administrations were well tolerated• EMEA criteria met for all treatment groups (fold titer, geometric mean titer, seroprotective rate)Moreover, 5-minute and 15-minute patch wear time produced similar immune response for all three strains and antibody titers remained elevated at six months after vaccination. The results demonstrate that MAPs obtained the same drug efficacy as the administration method through injection.Example 2Administration of SARS-CoV-2 circRNA Vaccine using MAPs

[0110] In this example, a patient is vaccinated against SARS-CoV-2. As described herein, the microneedle array patch includes a polymeric array containing hundreds of microneedles (the number of microneedles is dependent on the size of the array), each of which is coated with a droplet of vaccine.

[0111] The microneedle array was evaluated in the biocompatibility tests (i.e., cytotoxicity, sensitization, primary skin irritation, acute systemic toxicity, and ASTM Hemolysis). The results demonstrate the array was safe for administering an API or vaccine. Further, the constituents of the adhesive patch were evaluated for biocompatibility (cytotoxicity, irritation via intracutaneous reactivity test, primary skin irritation, and sensitization) with results meeting all test requirements. The skin contacting portions of the array patch applicator were also evaluated for biocompatibility (cytotoxicity, irritation, and sensitization) with the test articles meeting the test requirements.

[0112] Formulations for coating the microneedle arrays were manufactured by adding polymer excipients and sugars, either as dry powders or as highly concentrated solutions, to aliquots of a solution containing the cRNA vaccine. Water-soluble pharmaceutical grade polymers of varying molecular weights were chosen as polymer excipients based on previous development experience demonstrating these polymersPATENT EX1-005WOproduce robust coatings on the microneedle tips, while maintaining the stability of the vaccine that has been applied. Microneedle arrays are coated with the cRNA vaccine formulations using a dip coating and drying process, followed by a packaging step to ensure product stability.

[0113] To develop a safe and effective vaccine against SARS-CoV-2, a two-antigen strategy using the ciRNA™ platform was used. This involved creating circular RNA that encodes the complete SARS-CoV-2 spike protein (S), which has been shown to elicit strong neutralizing antibody responses in existing SARS-CoV-2 mRNA vaccines. The circRNA technology will facilitate prolonged spike antigen production, enhancing antibody durability. Following the latest WHO recommendations, the vaccine targets the JN.1 lineage, with flexibility to update the antigen sequence as needed.

[0114] In addition to a strong antibody response, a robust T-cell response is crucial for long-term protection against severe disease. To achieve this, a second ciRNA™ encoding the SARS-CoV-2 nucleoprotein (N) was incorporated, which is known to produce significant T-cell epitopes and a potent vaccine candidate. By combining these two ciRNA™s, a potent antibody response and a stable T-cell response was generated, resulting in a vaccine with a robust and durable immune profile.

[0115] The coated microneedle array is applied to the skin using an applicator, which ensures consistent delivery and ease of use by a medical professional or the patient. The coated microneedle array is held on the skin by an integrated adhesive patch. The droplets on the microneedles are delivered approximately 150μm to 350μm deep into the skin to an area with limited nerve endings, resulting in very low skin sensitivity. Once inserted into the skin, the API or vaccine formulation dissolves off the microneedles into the interstitial fluid. A typical delivery duration is between 30 seconds and five minutes, after which, the microneedle array patch is removed and disposed of. In this example, the patient wears the MAP for five minutes.

[0116] After 60 days, the patient was evaluated by her treating physician. A blood sample was analyzed to confirm that she was effectively immunized. In conclusion, the study demonstrated:PATENT EX1-005WO• a robust Immune response to SARS-CoV-2• the administration was well toleratedThe results demonstrate that MAPs obtained the same drug efficacy as the administration method through injection. Further, the patient expressed a preference for vaccination using MAPs rather than conventional intramuscular (IM) injection.* * *

[0117] Substances to be delivered using the microneedle patches or microneedle array patches (MAPs) described herein include bioactive substances, including pharmaceutical agents, medicaments, vaccines and the like. Substances may be in solid or liquid form, depending on formulation and delivery method. They can be delivered, inter alia, in the form of dry powders, gels, solutions, suspensions, and creams. Suitable formulations are familiar to those of skill in the art. Particularly preferred medicaments for delivery by the methods of the invention include vaccines, allergens and gene therapeutic agents.

[0118] In aspects, the space between microneedles is adjusted based on the intended use and / or active agent to administer.

[0119] In aspects, the microneedle length is adjusted based on the intended use and / or active agent to administer.

[0120] In aspects, the droplet volume / solids per microneedle is adjusted based on the intended use and / or active agent to administer.

[0121] In aspects, the droplet shape is adjusted based on the intended use and / or active agent to administer.

[0122] In aspects, the coated formulation is adjusted based on the intended use and / or active agent to administer.

[0123] In aspects, the number of microneedles is adjusted based on the intended use and / or active agent to administer.PATENT EX1-005WO

[0124] In aspects, each of the microneedles on a MAP is about 750 microns in length. Alternatively, each microneedle is about 325 microns, about 450 microns, about 500 microns, about 550 microns, about 600 microns, about 650 microns, about 700 microns, about 800 microns, about 850 microns, about 900 microns or about 950 microns in length.

[0125] In aspects, microneedles are separated from each other by a distance of about 650 microns. Alternatively, each microneedle can be separated by about 350, about 450 microns, about 500 microns, about 550 microns, about 600 microns, about 650 microns, about 700 microns, about 800 microns, about 850 microns, about 900 microns or about 950 microns.

[0126] Multiple doses can be administered to an individual in need thereof. Where multiple doses are administered over a period of time, an active agent is administered once a month to about once a year, from about once a year to once every 2 years, from about once every 2 years to once every 5 years, or from about once every 5 years to about once every 10 years, over a period of time. For example, a subject is administered over a period of from about 3 months to about 2 years, from about 2 years to about 5 years, from about 5 years to about 10 years, from about 10 years to about 20 years, or more than 20 years. The actual frequency of administration, and the actual duration of treatment, depends on various factors.

[0127] As an example, a subject at risk of an ailment can be immunized by administering an initial dose of a vaccine; and administering at least a second dose (a subsequent dose). Where two or more subsequent doses are administered, the subsequent dose(s) can be separated in time from each other by at least one month, at least 3 to 6 months, at least 6 months to 1 year, at least 1 year to 5 years, at least 5 years to 10 years, at least 10 years to 20 years, or more than 20 years.

[0128] The therapeutic methods described herein can include the step of administering drug product (i.e., a medicament or vaccine) at a pharmaceutically effective amount. The total daily dose should be determined through appropriate medical judgment by a physician and administered once or several times. The specificPATENT EX1-005WOtherapeutically effective dose level for any particular patient may vary depending on various factors well known in the medical art, including the kind and degree of the response to be achieved, concrete compositions according to whether other agents are used therewith or not, the patient’s age, body weight, health condition, gender, and diet, the time and route of administration, the secretion rate of the composition, the time period of therapy, other drugs used in combination or coincident with the composition disclosed herein, and like factors well known in the medical arts.

[0129] In one embodiment, the dose of the composition may be administered daily, semi-weekly, weekly, bi-weekly, or monthly. The period of treatment may be for a week, two weeks, a month, two months, four months, six months, eight months, a year, or longer. The initial dose may be larger than a sustaining dose. In one embodiment, the dose ranges from a weekly dose of at least 0.01 mg / kg, at least 0.25 mg / kg, at least 0.3 mg / kg, at least 0.5 mg / kg, at least 0.75 mg / kg, at least 1 mg / kg, at least 2 mg / kg, at least 3 mg / kg, at least 4 mg / kg, at least 5 mg / kg, at least 6 mg / kg, at least 7 mg / kg, at least 8 mg / kg, at least 9 mg / kg, at least 10 mg / kg, at least 15 mg / kg, at least 20 mg / kg, at least 25 mg / kg, or at least 30 mg / kg In one embodiment, a weekly dose may be at most 1.5 mg / kg, at most 2 mg / kg, at most 2.5 mg / kg, at most 3 mg / kg, at most 4 mg / kg, at most 5 mg / kg, at most 6 mg / kg, at most 7 mg / kg, at most 8 mg / kg, at most 9 mg / kg, at most 10 mg / kg, at most 15 mg / kg, at most 20 mg / kg, at most 25 mg / kg, or at most 30 mg / kg. In a particular aspect, the weekly dose may range from 5 mg / kg to 20 mg / kg. In an alternative aspect, the weekly dose may range from 10 mg / kg to 15 mg / kg.

[0130] The therapeutic agents in the pharmaceutical compositions can be formulated in a "therapeutically effective amount" or a "prophylactically effective amount". A "therapeutically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result. A therapeutically effective amount of the agent (i.e. a medicament or vaccine) may vary depending on the condition to be treated, the severity and course of the condition, the mode of administration, whether the agent is administered for preventive or therapeutic purposes, the bioavailability of the particular agent, the ability of the agent to elicit a desired response in the individual, previous therapy, the age, weight and sex of thePATENT EX1-005WOpatient, the patient's clinical history and response to the agent, discretion of the attending physician, etc. A therapeutically effective amount is also one in which any toxic or detrimental effects are outweighed by the therapeutically beneficial effects. A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result.

[0131] As a general proposition, a therapeutically effective amount or prophylactically effective amount of a medicament or vaccine will be administered in a range from about 1 ng / kg body weight to about 100 mg / kg body weight whether by one or more administrations. In a particular embodiment, the agent is administered in the range of from about 1 ng / kg body weight to about 10 mg / kg body weight, about 1 ng / kg body weight to about 1 mg / kg body weight, about 1 ng / kg body weight to about 100 g / kg body weight, about 1 ng / kg body weight to about 10 g / kg body weight, about 1 ng / kg body weight / day to about 1 g / kg body weight, about 1 ng / kg body weight to about 100 ng / kg body weight, about 1 ng / kg body weight to about 10 ng / kg body weight, about 10 ng / kg body weight to about 100 mg / kg body weight, about 10 ng / kg body weight to about 10 mg / kg body weight, about 10 ng / kg body weight to about 1 mg / kg body weight, about 10 ng / kg body weight / to about 100 g / kg body weight, about 10 ng / kg body weight to about 10 mg / kg body weight, about 10 ng / kg body weight to about 1 mg / kg body weight, 10 ng / kg body weight to about 100 ng / kg body weight / , about 100 ng / kg body weight to about 100 mg / kg body weight, about 100 ng / kg body weight to about 10 mg / kg body weight, about 100 ng / kg body weight to about 1 mg / kg body weight, about 100 ng / kg body weight to about 100 mg / kg body weight, about 100 ng / kg body weight to about 10 mg / kg body weight, about 100 ng / kg body weight to about 1 mg / kg body weight, about 1 mg / kg body weight to about 100 mg / kg body weight, about 1 mg / kg body weight to about 10 mg / kg body weight / day, about 1 mg / kg body weight to about 1 mg / kg body weight, about 1 mg / kg body weight to about 100 mg / kg body weight, about 1 mg / kg body weight to about 10 mg / kg body weight, about 10 mg / kg body weight to about 100 mg / kg body weight, about 10 mg / kg body weight to about 10 mg / kg body weight, about 10 mg / kg body weight to about 1 mg / kg body weight / day, about 10 mg / kg body weight to about 100 mg / kg body weight, about 100 mg / kg body weight / day to about 100 mg / kg body weight, about 100 mg / kg body weight / day to about 10 mg / kgPATENT EX1-005WObody weight, about 100 mg / kg body weight / day to about 1 mg / kg body weight, about 1 mg / kg body weight to about 100 mg / kg body weight, about 1 mg / kg body weight to about 10 mg / kg body weight, about 10 mg / kg body weight to about 100 mg / kg body weight / day.

[0132] In other embodiments, a medicament or vaccine is administered in the range of about 10 ng to about 100 ng per individual administration, about 10 ng to about 1 g per individual administration, about 10 ng to about 10 g per individual administration, about 10 ng to about 100 mg per individual administration, about 10 ng to about 1mg per individual administration, about 10 ng to about 10 mg per individual administration, about 10 ng to about 100 mg per individual administration, about 10 ng to about 1000 mg per injection, about 10 ng to about 10,000 mg per individual administration, about 100 ng to about 1 mg per individual administration, about 100 ng to about 10 mg per individual administration, about 100 ng to about 100 mg per individual administration, about 100 ng to about 1mg per individual administration, about 100 ng to about 10 mg per individual administration, about 100 ng to about 100 mg per individual administration, about 100 ng to about 1000 mg per injection, about 100 ng to about 10,000 mg per individual administration, about 1 mg to about 10 mg per individual administration, about 1 mg to about 100 mg per individual administration, about 1 mg to about 1 mg per individual administration, about 1 mg to about 10 mg per individual administration, about 1 mg to about 100 mg per individual administration, about 1 mg to about 1000 mg per injection, about 1 mg to about 10,000 mg per individual administration, about 10 mg to about 100 mg per individual administration, about 10 mg to about 1mg per individual administration, about 10 mg to about 10 mg per individual administration, about 10 mg to about 100 mg per individual administration, about 10 mg to about 1000 mg per injection, about 10 mg to about 10,000 mg per individual administration, about 100 mg to about 1mg per individual administration, about 100 mg to about 10 mg per individual administration, about 100 mg to about 100 mg per individual administration, about 100 mg to about 1000 mg per injection, about 100 mg to about 10,000 mg per individual administration, about 1mg to about 10 mg per individual administration, about 1 mg to about 100 mg per individual administration, about 1mg to about 1000 mg per injection, about 1mg to about 10,000 mg per individualPATENT EX1-005WOadministration, about 10 mg to about 100 mg per individual administration, about 10 mg to about 1000 mg per injection, about 10 mg to about 10,000 mg per individual administration, about 100 mg to about 1000 mg per injection, about 100 mg to about 10,000 mg per individual administration and about 1000 mg to about 10,000 mg per individual administration. The agent may be administered daily, every 2, 3, 4, 5, 6 or 7 days, or every 1, 2, 3 or 4 weeks.

[0133] In other particular embodiments, the amount of the a medicament or vaccine can be administered at a dose of about 0.0006 mg, 0.001 mg, 0.003 mg, 0.006 mg, 0.01 mg, 0.03 mg, 0.06 mg, 0.1 mg, 0.3 mg, 0.6 mg, 1 mg, 3 mg, 6 mg, 10 mg, 30 mg, 60 mg, 100 mg, 300 mg, 600 mg, 1000 mg, 2000 mg, 5000 mg or 10,000 mg. As expected, the dosage will be dependent on the condition, size, age and condition of the patient.

[0134] In embodiments, the amount of the a medicament or vaccine is administered at a therapeutically effective of a range from about 1 - 10 units, 10- 50 units, 10 - 100 units, 20 - 50 units, 20 - 100 units, 50 - 100 units, 50 - 150 units, 50 - 200 units, 50 -250 units, 50 - 300 units, 100 - 200 units, 100 - 250 units, 100 - 300 units, 100 - 350 units, 100 - 400 units, 200 - 250 units, 200 - 300 units, 200 - 350 units, 200 - 400 units, 300 - 400 units, 100 - 400 units, 300 - 500 units, or 400 - 500 units per treatment session.

[0135] A pharmaceutical composition disclosed herein is in an amount sufficient to allow customary administration to an individual. In aspects, a pharmaceutical composition disclosed herein can be, e.g., at least 5 mg, at least 10 mg, at least 15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at least 35 mg, at least 40 mg, at least 45 mg, at least 50 mg, at least 55 mg, at least 60 mg, at least 65 mg, at least 70 mg, at least 75 mg, at least 80 mg, at least 85 mg, at least 90 mg, at least 95 mg, or at least 100 mg of a pharmaceutical composition. In other aspects of this embodiment, a pharmaceutical composition disclosed herein may be, e.g., at least 5 mg, at least 10 mg, at least 20 mg, at least 25 mg, at least 50 mg, at least 75 mg, at least 100 mg, at least 200 mg, at least 300 mg, at least 400 mg, at least 500 mg, at least 600 mg, at least 700 mg, at least 800 mg, at least 900 mg, at least 1,000 mg, at least 1,100 mg, atPATENT EX1-005WOleast 1,200 mg, at least 1,300 mg, at least 1,400 mg, or at least 1,500 mg of a pharmaceutical composition. In yet other aspects, a pharmaceutical composition disclosed herein may be in the range of, e.g., about 5 mg to about 100 mg, about 10 mg to about 100 mg, about 50 mg to about 150 mg, about 100 mg to about 250 mg, about 150 mg to about 350 mg, about 250 mg to about 500 mg, about 350 mg to about 600 mg, about 500 mg to about 750 mg, about 600 mg to about 900 mg, about 750 mg to about 1,000 mg, about 850 mg to about 1,200 mg, or about 1,000 mg to about 1,500 mg. In still other aspects of this embodiment, a pharmaceutical composition disclosed herein may be in the range of, e.g., about 10 mg to about 250 mg, about 10 mg to about 500 mg, about 10 mg to about 750 mg, about 10 mg to about 1,000 mg, about 10 mg to about 1,500 mg, about 50 mg to about 250 mg, about 50 mg to about 500 mg, about 50 mg to about 750 mg, about 50 mg to about 1,000 mg, about 50 mg to about 1,500 mg, about 100 mg to about 250 mg, about 100 mg to about 500 mg, about 100 mg to about 750 mg, about 100 mg to about 1,000 mg, about 100 mg to about 1,500 mg, about 200 mg to about 500 mg, about 200 mg to about 750 mg, about 200 mg to about 1,000 mg, about 200 mg to about 1,500 mg, about 5 mg to about 1,500 mg, about 5 mg to about 1,000 mg, or about 5 mg to about 250 mg.

[0136] A pharmaceutical composition disclosed herein can comprise a solvent, emulsion or other diluent in an amount sufficient to dissolve a pharmaceutical composition disclosed herein. In other aspects of this embodiment, a pharmaceutical composition disclosed herein may comprise a solvent, emulsion or a diluent in an amount of, e.g., less than about 90% (v / v), less than about 80% (v / v), less than about 70% (v / v), less than about 65% (v / v), less than about 60% (v / v), less than about 55% (v / v), less than about 50% (v / v), less than about 45% (v / v), less than about 40% (v / v), less than about 35% (v / v), less than about 30% (v / v), less than about 25% (v / v), less than about 20% (v / v), less than about 15% (v / v), less than about 10% (v / v), less than about 5% (v / v), or less than about 1% (v / v). In other aspects of this embodiment, a pharmaceutical composition disclosed herein may comprise a solvent, emulsion or other diluent in an amount in a range of, e.g., about 1% (v / v) to 90% (v / v), about 1% (v / v) to 70% (v / v), about 1% (v / v) to 60% (v / v), about 1% (v / v) to 50% (v / v), about 1% (v / v) to 40% (v / v), about 1% (v / v) to 30% (v / v), about 1% (v / v) to 20% (v / v), about 1% (v / v) toPATENT EX1-005WO10% (v / v), about 2% (v / v) to 50% (v / v), about 2% (v / v) to 40% (v / v), about 2% (v / v) to 30% (v / v), about 2% (v / v) to 20% (v / v), about 2% (v / v) to 10% (v / v), about 4% (v / v) to 50% (v / v), about 4% (v / v) to 40% (v / v), about 4% (v / v) to 30% (v / v), about 4% (v / v) to 20% (v / v), about 4% (v / v) to 10% (v / v), about 6% (v / v) to 50% (v / v), about 6% (v / v) to 40% (v / v), about 6% (v / v) to 30% (v / v), about 6% (v / v) to 20% (v / v), about 6% (v / v) to 10% (v / v), about 8% (v / v) to 50% (v / v), about 8% (v / v) to 40% (v / v), about 8% (v / v) to 30% (v / v), about 8% (v / v) to 20% (v / v), about 8% (v / v) to 15% (v / v), or about 8% (v / v) to 12% (v / v).

[0137] The final concentration of a pharmaceutical composition (e.g., vaccine or medicament) disclosed herein can be of any concentration desired. In aspects, the final concentration of a pharmaceutical composition is a therapeutically effective amount. In other aspects, the final concentration of a pharmaceutical composition in a pharmaceutical composition may be, e.g., at least 0.00001 mg / mL, at least 0.0001 mg / mL, at least 0.001 mg / mL, at least 0.01 mg / mL, at least 0.1 mg / mL, at least 1 mg / mL, at least 10 mg / mL, at least 25 mg / mL, at least 50 mg / mL, at least 100 mg / mL, at least 200 mg / mL or at least 500 mg / mL. In other aspects of this embodiment, the final concentration of a pharmaceutical composition in a pharmaceutical composition may be in a range of, e.g., about 0.00001 mg / mL to about 3,000 mg / mL, about 0.0001 mg / mL to about 3,000 mg / mL, about 0.01 mg / mL to about 3,000 mg / mL, about 0.1 mg / mL to about 3,000 mg / mL, about 1 mg / mL to about 3,000 mg / mL, about 250 mg / mL to about 3,000 mg / mL, about 500 mg / mL to about 3,000 mg / mL, about 750 mg / mL to about 3,000 mg / mL, about 1,000 mg / mL to about 3,000 mg / mL, about 100 mg / mL to about 2,000 mg / mL, about 250 mg / mL to about 2,000 mg / mL, about 500 mg / mL to about 2,000 mg / mL, about 750 mg / mL to about 2,000 mg / mL, about 1,000 mg / mL to about 2,000 mg / mL, about 100 mg / mL to about 1,500 mg / mL, about 250 mg / mL to about 1,500 mg / mL, about 500 mg / mL to about 1,500 mg / mL, about 750 mg / mL to about 1,500 mg / mL, about 1,000 mg / mL to about 1,500 mg / mL, about 100 mg / mL to about 1,200 mg / mL, about 250 mg / mL to about 1,200 mg / mL, about 500 mg / mL to about 1,200 mg / mL, about 750 mg / mL to about 1,200 mg / mL, about 1,000 mg / mL to about 1,200 mg / mL, about 100 mg / mL to about 1,000 mg / mL, about 250 mg / mL to about 1,000 mg / mL, about 500 mg / mL to about 1,000 mg / mL, about 750 mg / mL toPATENT EX1-005WOabout 1,000 mg / mL, about 100 mg / mL to about 750 mg / mL, about 250 mg / mL to about 750 mg / mL, about 500 mg / mL to about 750 mg / mL, about 100 mg / mL to about 500 mg / mL, about 250 mg / mL to about 500 mg / mL, about 0.00001 mg / mL to about 0.0001 mg / mL, about 0.00001 mg / mL to about 0.001 mg / mL, about 0.00001 mg / mL to about 0.01 mg / mL, about 0.00001 mg / mL to about 0.1 mg / mL, about 0.00001 mg / mL to about 1 mg / mL, about 0.001 mg / mL to about 0.01 mg / mL, about 0.001 mg / mL to about 0.1 mg / mL, about 0.001 mg / mL to about 1 mg / mL, about 0.001 mg / mL to about 10 mg / mL, or about 0.001 mg / mL to about 100 mg / mL.

[0138] Aspects of the present specification disclose, in part, treating an individual suffering from an ailment. As used herein, the term "treating," refers to reducing or eliminating in an individual a clinical symptom of an ailment; or delaying or preventing in an individual the onset of a clinical symptom of an ailment. For example, the term "treating" can mean reducing signs / symptoms of an ailment, by, e.g., at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% at least 95%, or at least 100%. The actual symptoms associated with an ailment are well known and can be determined by a person of ordinary skill in the art. Those of skill in the art will know the appropriate symptoms or indicators associated with a specific type of ailment and will know how to determine if an individual is a candidate for treatment as disclosed herein.

[0139] In another aspect, a pharmaceutical composition disclosed herein reduces the severity of a symptom of a disorder associated an ailment. In aspects, a pharmaceutical composition disclosed herein reduces the severity of a symptom of a disorder associated with an ailment by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95%. In other aspects of this embodiment, a pharmaceutical composition disclosed herein reduces the severity of a symptom of a disorder associated with an ailment by, e.g., about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to aboutPATENT EX1-005WO100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70%.

[0140] In aspects of this embodiment, a therapeutically effective amount of a pharmaceutical composition disclosed herein reduces a symptom associated with an ailment or reduces the time of recovery by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 100%. In other aspects of this embodiment, a therapeutically effective amount of a pharmaceutical composition disclosed herein reduces a symptom associated with an ailment or reduces the time of recovery by, e.g., at most 10%, at most 15%, at most 20%, at most 25%, at most 30%, at most 35%, at most 40%, at most 45%, at most 50%, at most 55%, at most 60%, at most 65%, at most 70%, at most 75%, at most 80%, at most 85%, at most 90%, at most 95% or at most 100%. In yet other aspects of this embodiment, a therapeutically effective amount of a pharmaceutical composition disclosed herein reduces a symptom associated with an ailment or reduces the time of recovery by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 20%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0141] In yet other aspects of this embodiment, a therapeutically effective amount of a pharmaceutical composition disclosed herein generally is in the range of about 0.001 mg / kg to about 100 mg / kg and administered, for example, every 3, 5, 7, 10 or 14 days.PATENT EX1-005WOIn aspects of this embodiment, an effective amount of a pharmaceutical composition disclosed herein may be, e.g., at least 0.001 mg / kg, at least 0.01 mg / kg, at least 0.1 mg / kg, at least 1.0 mg / kg, at least 5.0 mg / kg, at least 10 mg / kg, at least 15 mg / kg, at least 20 mg / kg, at least 25 mg / kg, at least 30 mg / kg, at least 35 mg / kg, at least 40 mg / kg, at least 45 mg / kg, or at least 50 mg / kg and administered, for example, every 3, 5, 7, 10 or 14 days. In other aspects of this embodiment, an effective amount of a pharmaceutical composition disclosed herein may be in the range of, e.g., about 0.001 mg / kg to about 10 mg / kg, about 0.001 mg / kg / day to about 15 mg / kg, about 0.001 mg / kg to about 20 mg / kg, about 0.001 mg / kg to about 25 mg / kg, about 0.001 mg / kg to about 30 mg / kg, about 0.001 mg / kg to about 35 mg / kg, about 0.001 mg / kg to about 40 mg / kg, about 0.001 mg / kg to about 45 mg / kg, about 0.001 mg / kg to about 50 mg / kg, about 0.001 mg / kg to about 75 mg / kg, or about 0.001 mg / kg to about 100 mg / kg and administered, for example, every 3, 5, 7, 10 or 14 days. In yet other aspects of this embodiment, an effective amount of a pharmaceutical composition disclosed herein may be in the range of, e.g., about 0.01 mg / kg to about 10 mg / kg, about 0.01 mg / kg to about 15 mg / kg, about 0.01 mg / kg to about 20 mg / kg, about 0.01 mg / kg to about 25 mg / kg, about 0.01 mg / kg to about 30 mg / kg, about 0.01 mg / kg to about 35 mg / kg, about 0.01 mg / kg to about 40 mg / kg, about 0.01 mg / kg to about 45 mg / kg, about 0.01 mg / kg to about 50 mg / kg, about 0.01 mg / kg to about 75 mg / kg, or about 0.01 mg / kg to about 100 mg / kg and administered, for example, every 3, 5, 7, 10 or 14 days. In still other aspects of this embodiment, an effective amount of a pharmaceutical composition disclosed herein may be in the range of, e.g., about 0.1 mg / kg to about 10 mg / kg, about 0.1 mg / kg to about 15 mg / kg, about 0.1 mg / kg to about 20 mg / kg, about 0.1 mg / kg to about 25 mg / kg, about 0.1 mg / kg to about 30 mg / kg, about 0.1 mg / kg to about 35 mg / kg, about 0.1 mg / kg to about 40 mg / kg, about 0.1 mg / kg to about 45 mg / kg, about 0.1 mg / kg to about 50 mg / kg, about 0.1 mg / kg to about 75 mg / kg, or about 0.1 mg / kg to about 100 mg / kg and administered, for example, every 3, 5, 7, 10 or 14 days.

[0142] Dosing can be single dosage or cumulative (serial dosing), and can be readily determined by one skilled in the art. For example, treatment of an ailment can include a one-time administration of an effective dose of a pharmaceutical composition disclosed herein. Alternatively, treatment of an ailment can include multiplePATENT EX1-005WOadministrations of an effective dose of a pharmaceutical composition carried out over a range of time periods, such as, e.g., once daily, twice daily, trice daily, once every few days, or once weekly. The timing of administration can vary from individual to individual, depending upon such factors as the severity of an individual's symptoms. For example, an effective dose of a pharmaceutical composition disclosed herein can be administered to an individual once daily for an indefinite period of time, or until the individual no longer requires therapy. A person of ordinary skill in the art will recognize that the condition of the individual can be monitored throughout the course of treatment and that the effective amount of a pharmaceutical composition disclosed herein that is administered can be adjusted accordingly.

[0143] In aspects, a subject wears a MAP for a period of administration. The period of administration for a medicament or vaccine therapeutic can be, for example, about 1 minute, about 2 minutes, about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 45 minutes, about 60 minutes, or longer. Alternatively, the period of administration can be 1.5 hours, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, 16 hours or longer. Alternatively, the period of administration can be for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or more. In a further embodiment, a period of during which administration is stopped is for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months or more.

[0144] In aspects of this embodiment, an effective amount of a vaccine disclosed improves an immune response by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 100%. In other aspects of this embodiment, aPATENT EX1-005WOtherapeutically effective amount of a therapeutic disclosed herein reduces signs / symptoms of an ailment in an individual by, e.g., at most 10%, at most 15%, at most 20%, at most 25%, at most 30%, at most 35%, at most 40%, at most 45%, at most 50%, at most 55%, at most 60%, at most 65%, at most 70%, at most 75%, at most 80%, at most 85%, at most 90%, at most 95% or at most 100%. In yet other aspects of this embodiment, a therapeutically effective amount of a therapeutic disclosed herein signs / symptoms of an ailment in an individual with the disease by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 20%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0145] The present specification also provides a pharmaceutical composition for the administration to a subject using MAPs. The pharmaceutical composition disclosed herein may further include a pharmaceutically acceptable carrier, excipient, or diluent. As used herein, the term "pharmaceutically acceptable" means that the composition is sufficient to achieve the therapeutic effects without deleterious side effects, and may be readily determined depending on the type of the diseases, the patient's age, body weight, health conditions, gender, and drug sensitivity, administration route, administration mode, administration frequency, duration of treatment, drugs used in combination or coincident with the composition disclosed herein, and other factors known in medicine.

[0146] The composition may be used by blending with a variety of pharmaceutically acceptable carriers such as physiological saline or organic solvents. In order to increase the stability or absorptivity, carbohydrates such as glucose, sucrose or dextrans, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers may be used.

[0147] The administration dose and frequency of the pharmaceutical compositionPATENT EX1-005WOdisclosed herein are determined by the type of active ingredient, together with various factors such as the disease to be treated, administration route, patient's age, gender, and body weight, and disease severity.

[0148] The total effective dose of the compositions disclosed herein may be administered to a patient in a single dose, or may be administered for a long period of time in multiple doses according to a fractionated treatment protocol. In the pharmaceutical composition disclosed herein, the content of active ingredient may vary depending on the disease severity. Preferably, the total daily dose of the agent disclosed herein may be approximately 0.0001 μg to 500 mg per 1 kg of body weight of a patient. However, the effective dose of the peptide is determined considering various factors including patient's age, body weight, health conditions, gender, disease severity, diet, and secretion rate, in addition to administration route and treatment frequency of the pharmaceutical composition. In view of this, those skilled in the art may easily determine an effective dose suitable for the particular use of the pharmaceutical composition disclosed herein. The pharmaceutical composition disclosed herein is not particularly limited to the formulation, and administration route and mode, as long as it shows suitable effects.

[0149] Moreover, the pharmaceutical composition may be administered alone or in combination or coincident with other pharmaceutical formulations showing prophylactic or therapeutic efficacy.

[0150] Given the teachings and guidance provided herein, those skilled in the art will understand that a formulation described herein can be equally applicable to many types of biopharmaceuticals, including those exemplified, as well as others known in the art. Given the teachings and guidance provided herein, those skilled in the art also will understand that the selection of, for example, type(s) or and / or amount(s) of one or more excipients, surfactants and / or optional components can be made based on the chemical and functional compatibility with the biopharmaceutical to be formulated and / or the mode of administration as well as other chemical, functional, physiological and / or medical factors well known in the art. For example, non-reducing sugars exhibitPATENT EX1-005WOfavorable excipient properties when used with polypeptide biopharmaceuticals compared to reducing sugars. Accordingly, exemplary formulations are exemplified further herein with reference to polypeptide biopharmaceuticals. However, the range of applicability, chemical and physical properties, considerations and methodology applied to polypeptide biopharmaceutical can be similarly applicable to biopharmaceuticals other than polypeptide biopharmaceuticals.

[0151] In various embodiments, a formulation can include combinations of bioactive agents (such as viruses, proteins, antibodies, peptides and the like as described herein) in the formulation. For example, a formulation as described herein can include a single bioactive agent for treatment of one or more conditions. A formulation as described herein also can include, in an embodiment, two or more different bioactive agents for a single or multiple conditions. Use of multiple bioactive agents in a formulation can be directed to, for example, the same or different indications. Similarly, in another embodiment, multiple bioactive agents can be used in a formulation to treat, for example, both a pathological condition and one or more side effects caused by the primary treatment. In a further embodiment, multiple bioactive agents also can be included, in a formulation as described herein to accomplish different medical purposes including, for example, simultaneous treatment and monitoring of the progression of the pathological condition. In an additional embodiment, multiple, concurrent therapies such as those exemplified herein as well as other combinations well known in the art are particularly useful for patient compliance because a single formulation can be sufficient for some or all suggested treatments and / or diagnosis. Those skilled in the art will know those bioactive agents that can be admixed for a wide range of combination therapies. Similarly, in various embodiments, a formulation can be used with a small molecule drug and combinations of one or more bioactive agents together with one or more small molecule pharmaceuticals. Therefore, in various embodiments a formulation is provided containing 1, 2, 3, 4, 5 or 6 or more different bioactive agents, as well as, for one or more bioactive agents combined with one or more small molecule pharmaceuticals.

[0152] In various embodiments, a formulation can include one or morePATENT EX1-005WOpreservatives and / or additives known in the art. Similarly, a formulation can further be formulated, without limitation, into any of various known delivery formulations. For example, in an embodiment, a formulation can include, surfactants, adjuvant, biodegradable polymers, hydrogels, etc., such optional components, their chemical and functional characteristics are known in the art. Similarly known in the art are formulations that facilitate rapid, sustained or delayed release of the bioactive agents after administration. A formulation as described can be produced to include these or other formulation components known in the art.

[0153] The composition can therefore be administered as a single dose, or as two or more doses (which may or may not contain the same amount of the desired molecule) over time, or as a continuous infusion via an implantation device or catheter. Further refinement of the appropriate dosage is routinely made by those of ordinary skill in the art and is within the ambit of tasks routinely performed by them. Appropriate dosages may be ascertained through use of appropriate dose-response data. In various embodiments, the bioactive agents in formulations described herein can, without limitation, be administered to patients throughout an extended time period, such as chronic administration for a chronic condition. The composition can be a solid, a semi-solid or an aerosol and a pharmaceutical compositions is formulated as a tablet, geltab, lozenge, orally dissolved strip, capsule, syrup, oral suspension, emulsion, granule, sprinkle or pellet.

[0154] The doses of the medicament or vaccine can be expressed in units per injection. For example, a medicament or vaccine can have a specific activity of about 10,000 units / mg to about 25,000 units / mg, or about 15,000 units / mg, or about 17,500 units / mg, or about 20,000 units / mg, or about 22,500 units / mg, or about 10,000 units / 0.58 mg, or 17,241 units / mg wherein “mg” refers to the amount of units present in a composition (as distinct from excipients and other constituents). Accordingly, the present invention contemplates injecting about 500 units to about 50,000 units per treatment session, or about 10,000 units to about 25,000 units per treatment session.

[0155] In another embodiment, the dose of a medicament or vaccine per injection isPATENT EX1-005WOabout 50 units to about 2,500 units, or about 85 units to about 2,000 units, or about 150 units to about 1,750 units, or about 200 units to about 1,500 units, or about 300 units to about 1,250 units, or about 500 units to about 1,000 units.

[0156] Packaging and instruments for administration may be determined by a variety of considerations, such as, without limitation, the volume of material to be administered, the conditions for storage, whether skilled healthcare practitioners will administer or patient self-compliance, the dosage regime, the geopolitical environment (e.g., exposure to extreme conditions of temperature for developing nations), and other practical considerations.

[0157] Compositions in accordance with embodiments described herein have desirable properties, such as desirable solubility, viscosity, syringeability and stability. Lyophilates in accordance with embodiments described herein have desirable properties, as well, such as desirable recovery, stability and reconstitution.

[0158] In an embodiment, the pH of the pharmaceutical formulation is at least about 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, or 9.

[0159] In an embodiment, the pH of the pharmaceutical formulation is from about 3 to about 9, about 4 to about 19, about 5 to about 9, about 6 to about 8, about 6 to about 7, about 6 to about 9, about 5 to about 6, about 5 to about 7, about 5 to about 8, about 4 to about 9, about 4 to about 8, about 4 to about 7, about 4 to about 6, about 4 to about 5, about 3 to about 8, about 3 to about 7, about 3 to about 6, about 3 to about 5, about 3 to about 4, about 7 to about 8, about 7 to about 9, about 7 to about 10.

[0160] Certain embodiments of the present invention are described herein, including the best mode known to the inventors for carrying out the invention. Of course, variations on these described embodiments will become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventor expects skilled artisans to employ such variations as appropriate, and the inventors intend for the present invention to be practiced otherwise than specifically described herein.PATENT EX1-005WOAccordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described embodiments in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

[0161] Groupings of alternative embodiments, elements, or steps of the present invention are not to be construed as limitations. Each group member may be referred to and claimed individually or in any combination with other group members disclosed herein. It is anticipated that one or more members of a group may be included in, or deleted from, a group for reasons of convenience and / or patentability. When any such inclusion or deletion occurs, the specification is deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.

[0162] As used herein, the term “about” means that the characteristic, item, quantity, parameter, property, or term so qualified encompasses a range of plus or minus ten percent above and below the value of the stated characteristic, item, quantity, parameter, property, or term. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and attached claims are approximations that may vary. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical indication should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and values setting forth the broad scope of the invention are approximations, the numerical ranges and values set forth in the specific examples are reported as precisely as possible. Any numerical range or value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Recitation of numerical ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate numerical value falling within the range. Unless otherwise indicated herein, each individual value of a numerical range is incorporated into the present specification as if it were individuallyPATENT EX1-005WOrecited herein.

[0163] The terms “a,” “an,” “the” and similar referents used in the context of describing the present invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein is intended merely to better illuminate the present invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the present specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0164] Specific embodiments disclosed herein may be further limited in the claims using consisting of or consisting essentially of language. When used in the claims, whether as filed or added per amendment, the transition term “consisting of” excludes any element, step, or ingredient not specified in the claims. The transition term “consisting essentially of” limits the scope of a claim to the specified materials or steps and those that do not materially affect the basic and novel characteristic(s).Embodiments of the present invention so claimed are inherently or expressly described and enabled herein.

[0165] Groupings of alternative embodiments, elements, or steps of the present invention are not to be construed as limitations. Each group member may be referred to and claimed individually or in any combination with other group members disclosed herein. It is anticipated that one or more members of a group may be included in, or deleted from, a group for reasons of convenience and / or patentability. When any such inclusion or deletion occurs, the specification is deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.

[0166] All patents, patent publications, and other publications referenced and identified in the present specification are individually and expressly incorporated hereinPATENT EX1-005WOby reference in their entirety for the purpose of describing and disclosing, for example, the compositions and methodologies described in such publications that might be used in connection with the present invention. These publications are provided solely fortheir disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicants and does not constitute any admission as to the correctness of the dates or contents of these documents.

[0167] In closing, it is to be understood that although aspects of the present specification are highlighted by referring to specific embodiments, one skilled in the art will readily appreciate that these disclosed embodiments are only illustrative of the principles of the subject matter disclosed herein. Therefore, it should be understood that the disclosed subject matter is in no way limited to a particular methodology, protocol, and / or reagent, etc., described herein. As such, various modifications or changes to or alternative configurations of the disclosed subject matter can be made in accordance with the teachings herein without departing from the spirit of the present specification. Lastly, the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention, which is defined solely by the claims. Accordingly, the present invention is not limited to that precisely as shown and described.

Claims

PATENT EX1-005WOCLAIMSWhat is claimed is:

1. A microneedle array patch (MAP) comprising:a) a base;b) an adhesive region; andb) a plurality of needles on a surface of the base,wherein the needles comprise a formulation with an active agent.

2. The MAP of claim 1, wherein each of the plurality of microneedles is about 750 microns in length.

3. The MAP of claim 1, wherein the plurality of microneedles are separated from each other by about 650 microns.

4. The MAP of claim 1, wherein each of the plurality of microneedles is a solid microneedle.

5. The MAP of claim 1, wherein each of the plurality of microneedles is a coated microneedle.

6. The MAP of claim 1, wherein each of the plurality of microneedles is a hollow microneedle.

7. The MAP of claim 1, wherein each of the plurality of microneedles is a dissolvable microneedle.

8. The MAP of claim 1, wherein each of the plurality of microneedles is a hydrogel-forming microneedle.

9. The MAP of claim 1, wherein the active agent is a small molecule therapeutic.

10. The MAP of claim 9, wherein the small molecule therapeutic is selected from aspirin, acetaminophen, antihistamines, ibuprofen and penicillinPATENT EX1-005WO11. The MAP of claim 1, wherein the active agent is large molecule therapeutic.

12. The MAP of claim 11, wherein large molecule therapeutic is selected from a protein, a peptide, a nucleotide sequence, a monoclonal antibody, a vaccine, and a polysaccharide.

13. The MAP of claim 12, wherein the vaccine is selected from Covid vaccine, flu vaccine, Lyme disease vaccine, rabies vaccine, measles vaccine, mumps vaccine, chicken pox vaccine, small pox vaccine, hepatitis vaccine, pertussis vaccine, rubella vaccine, diphtheria vaccine, encephalitis vaccine, yellow fever vaccine, recombinant protein vaccine, DNA vaccines, RNA vaccines, polio vaccine, therapeutic cancer vaccine, herpes vaccine, pneumococcal vaccine, meningitis vaccine, whooping cough vaccine, tetanus vaccine, typhoid fever vaccine, cholera vaccine, tuberculosis vaccine, and combinations thereof.

14. The MAP of claim 12, wherein the vaccine is an mRNA vaccine.

15. The MAP of claim 14, wherein the mRNA vaccine is a circular RNA vaccine.

16. A method of administering a medicament to a subject, the method comprising applying the MAP of claim 1 to an area of skin of a subject.

17. The method of claim 16, wherein an applicator is used in the step of applying the Map to an area of skin of the subject.

18. The method of claim 16, wherein each of the microneedles penetrates about 150µm to 450µm into the skin of the subject.

19. The method of claim 16, wherein about 10µg to 30µg of circRNA is administered to the subject.

20. A method of intradermal administration of a vaccine to a subject, the method comprising steps of:a) providing a microneedle array patch (MAP), andb) applying the MAP to an area of skin of the subject,PATENT EX1-005WOwherein the MAP comprises a plurality of needles on a surface,wherein each of the plurality of needles comprises a vaccine composition.

21. The method of claim 20, wherein the vaccine composition elicits an immune response against a bacteria.

22. The method of claim 20, wherein the vaccine composition elicits an immune response against a virus.

23. The method of claim 20, wherein the vaccine composition elicits an immune response against a parasite.

24. The method of claim 20, wherein the vaccine composition elicits an immune response against a fungus.

25. The method of claim 20, wherein the vaccine composition elicits an immune response against a cancer.

26. The method of claim 20, wherein the vaccine composition comprises a 25 µg to 150 µg dose of circular RNA.

27. The method of claim 20, wherein the vaccine composition comprises Covid vaccine, flu vaccine, Lyme disease vaccine, rabies vaccine, measles vaccine, mumps vaccine, chicken pox vaccine, small pox vaccine, hepatitis vaccine, pertussis vaccine, rubella vaccine, diphtheria vaccine, encephalitis vaccine, yellow fever vaccine, recombinant protein vaccine, DNA vaccines, RNA vaccines, polio vaccine, therapeutic cancer vaccine, herpes vaccine, pneumococcal vaccine, meningitis vaccine, whooping cough vaccine, tetanus vaccine, typhoid fever vaccine, cholera vaccine, tuberculosis vaccine, and combinations thereof.

28. The method of claim 20, wherein the vaccine composition comprises circular RNA.

29. The method of claim 20, wherein the vaccine composition elicits an immune response against SARS-CoV-2 spike protein (S).PATENT EX1-005WO30. The method of claim 29, wherein the plurality of needles comprises a second vaccine composition, where the second vaccine composition elicits an immune response against SARS-CoV-2 nucleoprotein (N).

31. A microneedle array patch comprising:(a) a backing substrate;(b) a plurality of microneedles extending from the backing substrate, each microneedle having a length of 300 - 1000 pm; and(c) a coating disposed on at least a portion of the microneedles,wherein the coating comprises:(i) a lipid nanoparticle encapsulating a circular RNA,(ii) sucrose, and(iii) a polymer selected from polyvinyl alcohol (PVA) and polyvinylpyrrolidone (PVP),wherein the circular RNA comprises:(A) an internal ribosome entry site (IRES);(B) an open reading frame encoding a therapeutic protein; and(C) a covalently closed circular backbone.

32. The microneedle array patch of claim 31, wherein the microneedles have a length of about 750 pm.

33. The microneedle array patch of claim 31, wherein the lipid nanoparticle comprises an ionizable lipid, cholesterol, a phospholipid and a PEG-lipid.

34. The microneedle array patch of claim 31, wherein the circular RNA encodes a viral antigen.

35. The microneedle array patch of claim 34, wherein the viral antigen is a coronavirus spike protein.PATENT EX1-005WO36. The microneedle array patch of claim 31, wherein the circular RNA encodes a tumor-associated antigen.

37. The microneedle array patch of claim 31, wherein the coating is solidified on the microneedles following application of an aqueous formulation.

38. A pharmaceutical formulation for coating a microneedle array patch, comprising:(a) a lipid nanoparticle encapsulating a circular RNA comprising an IRES and an open reading frame encoding a therapeutic protein;(b) sucrose;(c) a polymer selected from PVA and PVP; and(d) water.

39. A method of delivering a therapeutic protein to a subject, comprising applying the microneedle array patch of claim 31 to the skin of the subject for a period of 5 to 30 minutes,wherein the circular RNA is delivered into dermal cells and expressed to produce the therapeutic protein.

40. The method of claim 39, wherein the therapeutic protein is a vaccine antigen and the method induces an immune response in the subject.

41. The method of claim 40, wherein the immune response comprises production of antigen-specific antibodies.

42. The method of claim 39, wherein the therapeutic protein is a tumor-associated antigen and the method induces an anti-tumor immune response.

43. The microneedle array patch of claim 31, wherein the coating retains at least 80% circular RNA integrity after storage at 25°C for 30 days.PATENT EX1-005WO44. The method of claim 39, wherein expression of the therapeutic protein is detected in dermal tissue and in at least one regional lymph node.

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