Transdermal delivery of large drugs

Abstract

Providing transdermal delivery of large drugs. [Solution] Methods, compositions, and devices for enhancing transdermal delivery of large drugs. The present disclosure provides improved techniques for transdermal delivery of drugs of interest. In particular, the present disclosure provides insight that certain advantages can be achieved when microneedle processing technology is combined with emulsion technology for transdermal delivery of drugs of interest. Alternatively or additionally, in some embodiments, the present disclosure demonstrates that surprising enhancements for transdermal delivery of large molecular structures can be achieved when microneedle processing technology is utilized.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Patent Application No. 62 / 424,937, filed on 21 November 2016, which is incorporated herein by reference in its entirety. [Background technology]

[0002] Significant funds are being invested in the development of effective transdermal delivery technologies. Those skilled in the art are well aware of the challenges associated with achieving effective transdermal delivery, particularly for large-cap drugs. As molecular size increases, transdermal penetration decreases to the point of being negligible or even nonexistent. Even in those cases where some penetration occurs, large-cap drugs tend to be degraded and become biologically inactive. Novel and effective technologies for the effective transdermal administration of such drugs, which are large-cap drugs and are also susceptible to degradation, particularly botulinum toxin and antibody drugs, remain in need. [Overview of the project] [Means for solving the problem]

[0003] Transdermal administration continues to be a subject of research in attempts to provide an alternative route of drug administration that does not involve the undesirable consequences associated with injection and oral delivery. For example, needles often cause local pain, bleeding, and bruising, potentially exposing patients to infectious diseases; oral administration can suffer from insufficient bioavailability of the drug due to the highly acidic environment of the patient's stomach. In some embodiments, transdermal delivery has a more uniform, regular, and / or consistent pharmacokinetic profile compared to other routes of administration.

[0004] While transdermal drug delivery offers many advantages, it also presents numerous logistical challenges. Only a limited number of drugs have been shown to be deliverable via this route. Transdermal delivery of active drugs, including but not limited to hydrophilic molecules, large molecular structures (e.g., exceeding several hundred daltons), gene therapies, and vaccines, remains challenging. (Prusnitz, MR & Langer, R. “Transdermal drug delivery,” Nat Biotechnol. 26(11):1261-1268(2008)).

[0005] This disclosure provides improved techniques for the transdermal delivery of target drugs. In particular, this disclosure provides insight that certain advantages can be achieved when microneedle processing techniques are combined with emulsion techniques for the transdermal delivery of target drugs. Alternatively or additionally, in some embodiments, this disclosure demonstrates that remarkable advances can be achieved for the transdermal delivery of large molecular structures when microneedle processing techniques are utilized.

[0006] Various microneedling techniques have been developed that may be useful for administering a specific drug of interest. Microneedling can avoid certain drawbacks often associated with the use of larger needles (e.g., using standard injection techniques), such as pain and / or the amount of bleeding. Microneedling techniques may utilize one or more hollow or solid microneedles (e.g., an array thereof). The drug of interest is placed inside (e.g., if the microneedle is hollow and / or if the drug is incorporated into the microneedle material) or on (i.e., on the surface of the microneedle(s)) and / or can be applied to a skin site before, during, or after microneedling of that site. The drug inside or on the microneedle may be released, for example, by diffusion or release from the microneedle, or by rupture and / or fragmentation of the microneedle material after application to the site.

[0007] In some embodiments, the disclosure provides strategies for using microneedle treatment to “prepare” (and specifically, pre-prepare the skin before administration of a major drug) the skin to which a transdermal product has been, is being, or will be applied. Studies analyzing the transdermal delivery of small molecules (specifically, short hydrophilic peptides having molecular weights in the range of 400–1000 Da) have shown that “skin penetration of peptides depends on their molecular weight and decreases as the molecular weight increases” (Zhang, S., et al., “Enhanced delivery Having found that such a strategy is likely only useful for drugs with low molecular weights ("Acta Pharmaceutica Sinica B.4(1):100-104(2014)"), this disclosure provides the insight that, surprisingly, such microneedle preparation can offer significant benefits in facilitating the transdermal delivery of large drugs (e.g., those with molecular weights of approximately 100 kDa or more).

[0008] Therefore, prior to this disclosure, those skilled in the art would have understood from the literature that it would not be expected that microneedle preparation of the skin would facilitate the transdermal delivery of large drugs. This disclosure, surprisingly, demonstrates that microneedle preparation of the skin can significantly facilitate the transdermal delivery of drugs such as botulinum toxin, which has a molecular weight of approximately 150,000 kDa. Standard antibodies have a similar molecular weight.

[0009] This disclosure particularly demonstrates that microneedle treatment techniques (e.g., microneedle preparation of the skin) can significantly enhance the transdermal delivery of large-capacity drugs in emulsion compositions (e.g., macroemulsion compositions and / or nanoemulsion compositions). As illustrated, for example, pre-preparation of the skin by applying microneedles before any administration of the relevant large-capacity drug (botulinum toxin) surprisingly enhanced the delivery of the large-capacity drug through the skin. Specific examples included herein document such enhanced delivery under various conditions and / or circumstances (e.g., different skin sites, number of applications, etc.). Those skilled in the art will recognize other types (e.g., with respect to application site, number of doses, etc.) that fall within the scope of this disclosure.

[0010] The specific nanoemulsion compositions covered include water-in-oil and oil-in-water nanoemulsions characterized by droplet sizes ranging from about 10 nm to about 300 nm in diameter, an oil-to-aqueous dispersion ratio ranging from about 0.01:1 to about 20:1, an oil-to-surfactant ratio ranging from about 0.1 to about 40, and / or a zeta potential ranging from about -80 mV to about +80 mV (see, for example, the descriptions of nanoemulsion compositions in one or more of PCT / US2006 / 26918; PCT US06 / 46236; PCT / US2012 / 22276; and PCT / US2012 / 22279 (each disclosure being incorporated herein by reference in whole)).

[0011] The findings presented herein are particularly surprising, given reports that transdermal delivery of solid nanoparticles of a size comparable to that of droplets in the nanoemulsion compositions used herein (e.g., 10⁵ ± 2.92 nm) does not effectively deliver (or facilitate) even small molecule drugs transdermally through the skin. For example, Gomaa et al. described a study in which a solution of rhodamine dye (molecular weight 479 Da) encapsulated in PLGA nanoparticles was applied to skin pre-prepared by microneedle treatment to evaluate skin penetration. Gomaa, Y., et al, “Effect of microneedle treatment” See “On the skin permeation of a nanoencapsulated dye.” J Pharm Pharmacol. 2012 November;64(11):1592-1602. The data showed that very small amounts of dye began to penetrate the skin after 6 hours of continuous application; no significant increase in penetration was observed until the skin was treated continuously for 24 hours. The researchers explained that “there is a new consensus that NPs [nanoparticles] cannot normally penetrate the stratum corneum but can be well deposited in hair follicles.” Thus, before this disclosure, those skilled in the art would expect that the use of microneedle treatment techniques using nanosized vehicles would not be able to effectively deliver even small molecule drugs (e.g., rhodamine dye) transdermally; and that delivery of large molecules would certainly be impossible. However, this disclosure demonstrates that microneedle treatment can significantly facilitate the transdermal delivery of large molecules, especially when used in conjunction with nanoemulsion systems.

[0012] In particular, this disclosure demonstrates that microneedling techniques can facilitate transdermal delivery (e.g., large drugs from macroemulsion or nanoemulsion compositions) when no other disintegrants are used (i.e., no chemical penetration enhancers are used, and no other techniques are used to disintegrate or puncture skin structures). Previous studies of transdermal delivery of drugs roughly the size of botulinum toxin (i.e., about 150 kDa) using microneedles have reported that delivery is unsuccessful unless additional treatment is applied to disintegrate the skin. For example, U.S. Patent Publication 2010 / 0196445 reports that botulinum toxin is not effectively delivered from pre-coated microneedles unless skin digestive enzymes are also applied to disintegrate skin structures at the microneedling site.

[0013] In some embodiments, the Disclosure provides techniques for achieving enhanced transdermal delivery of large-capacity drugs (e.g., botulinum toxin, antibodies, etc.) by utilizing microneedle processing techniques without the additional use of penetration enhancers. Alternatively or additionally, in some embodiments, the Disclosure provides techniques for achieving enhanced transdermal delivery of large-capacity drugs (e.g., botulinum toxin, antibodies, etc.) by utilizing microneedle processing techniques without any other disintegrating measures. Thus, the techniques provided can achieve effective delivery without the inflammation, irritation, and / or allergic reactions that often occur with the use of skin-disintegrating agents.

[0014] Alternatively or additionally, this disclosure identifies the causes of problems with certain conventional approaches to relevant large-cap drugs, particularly large protein drugs (e.g., botulinum toxin, antibodies, etc.), in or on microneedle structures. Typically, such conventional relevant strategies utilize a solution of the relevant drug that can be applied to a microneedle and air-dried. Such a strategy was used to coat a microneedle with botulinum toxin in U.S. Patent Publication 2010 / 0228225. U.S. Patent Publication 2017 / 0209553 describes a microneedle array in which botulinum is loaded into the needles. This disclosure understands that botulinum coating or loading materials produced thereby are not stable and therefore not commercially viable when used to manufacture products. Indeed, even if such liquids are prepared from powder materials, this disclosure understands that for many large-cap drugs (e.g., botulinum toxin), powders and other solid materials not formed by a freeze-drying process can be highly unstable. For example, according to Johnson, E., et al., "Botulinum toxin is highly susceptible to denaturation due to surface denaturation, heat, and alkaline conditions. Lyophilization or freeze-drying of botulinum toxin is the most economically rational and practical method of distributing the product in a stable and readily usable form for clinicians." U.S. Patent No. 5,512,547. Similarly, such an approach would not work for the administration of therapeutic antibodies, which have their own stability and storage challenges. This disclosure provides insight that the use of emulsion compositions described herein (e.g., nanoemulsion compositions in some embodiments, and / or macroemulsion compositions in some embodiments) can protect or otherwise improve the stability of large drugs, particularly large protein drugs (specifically, including botulinum toxin and / or antibody drugs), in relation to microneedles.

[0015] The present disclosure provides surprisingly effective techniques for the transdermal delivery of large drugs. In particular, the present disclosure teaches that the transdermal delivery of such drugs can be significantly enhanced through the use of a given microneedle treatment technique. In some embodiments, the present disclosure teaches that particularly advantageous results are achieved when the microneedle treatment technique is combined with an emulsion composition (e.g., in some embodiments, a nanoemulsion composition, and / or in some embodiments a macroemulsion composition). In some embodiments, the microneedle treatment technique is combined with a lotion, cream, or liquid composition that can later become or include the emulsion composition (e.g., in some embodiments with a nanoemulsion embodiment, and / or in some embodiments with a macroemulsion composition). In some embodiments, the techniques provided do not utilize skin disruption techniques such as chemical penetration enhancers.

Brief Description of the Drawings

[0016] [Figure 1] The results of the rat study of the MSC ("microneedle skin conditioning") effect on the bioavailability of the botulinum nanoemulsion formulation are illustrated by the finger abduction scoring. [Figure 2] The survival rate of the rat study of the MSC effect on the bioavailability of the botulinum nanoemulsion formulation is illustrated.

Modes for Carrying Out the Invention

[0017] Definitions Unless otherwise apparent from the context, in this application: (i) the term "a" can be understood to mean "at least one"; (ii) the term "or" can be understood to mean "and / or"; (iii) the terms "comprising" and "including" can be understood to include the recited element or step, whether or not it is presented by itself or in combination with one or more additional elements or steps; (iv) the terms "about" and "approximately" can be understood to allow for a standard deviation as would be understood by one of ordinary skill in the art; (v) when ranges are provided, endpoints are included.

[0018] Abrasion: As used herein, the term "abrasion" refers to any means of modifying, disintegrating, removing, or destroying the uppermost layer of the skin. In some embodiments, abrasion refers to mechanical means of modifying, disintegrating, removing, or destroying the uppermost layer of the skin. In some embodiments, abrasion refers to chemical means of modifying, disintegrating, removing, or destroying the uppermost layer of the skin. By way of example, but not limitation, agents such as scrubs, microparticles (e.g., magnesium or aluminum particles), acids (e.g., alpha-hydroxy acids or beta-hydroxy acids), and alcohols can cause abrasion. In general, penetration enhancers such as those described by, for example, Donovan (e.g., U.S. Publication Nos. 2004 / 009180 and 2005 / 175636, and PCT Publication No. WO04 / 06954), and Graham (e.g., U.S. Patent No. 6,939,852 and U.S. Publication No. 2006 / 093624) are expected to cause abrasion. Of course, one of ordinary skill in the art will understand that a particular agent may cause abrasion at one concentration or in combination with one or more other agents, but may not cause abrasion under different circumstances. Thus, whether a particular material is an "abrasive agent" depends on the circumstances. Abrasion can be readily evaluated by one of ordinary skill in the art, for example, by observation of skin redness or irritation and / or histological examination of the skin showing changes, disintegration, removal, or erosion of the stratum corneum.

[0019] Administration: As used herein, the term “administration” typically refers to the administration of a composition to a subject or system. Those skilled in the art will recognize, in appropriate circumstances, the various routes that may be used for administration to a subject, e.g., a human. For example, in some embodiments, administration may be intraocular, oral, parenteral, topical, etc. In some embodiments, administration may be intrabronchial (e.g., by intrabronchial infusion), oral cavity, percutaneous (e.g., one or more of the following in the dermis: topical, intradermal, intercutaneous, percutaneous, etc., or may include them), intestinal, intraarterial, intradermal, gastric, intramedullary, intramuscular, intranasal, intraperitoneal, intramedullary, intravenous, intraventricular, intraspecific organ (e.g., intrahepatic), mucosa, nasal, oral, rectal, subcutaneous, sublingual, topical, trachea (e.g., by intratracheal infusion), vaginal, vitreous, etc. In some embodiments, administration may include medication that is intermittent (e.g., multiple doses spaced apart in time) and / or periodic (e.g., individual doses separated by a common period). In some embodiments, administration may include continuous medication (e.g., perfusion) over at least a selected period.

[0020] Drugs: Generally, the term “drugs” as used herein may be used to refer to compounds or entities of any chemical classification, including, for example, polypeptides, nucleic acids, saccharides, lipids, small molecules, metals, or combinations or complexes thereof. Where appropriate, as will be apparent to those skilled in the art from the context, the term may be used to refer to cells or organisms, or fractions, extracts, or components thereof, or entities containing them. Alternatively or additionally, as will be apparent from the context, the term may be used to refer to natural products in that they are found in nature and / or obtained from nature. In some cases, as will also be apparent from the context, the term may be used to refer to one or more artificial entities in that they are designed, devised, and / or produced by human hands and / or not found in nature. In some embodiments, drugs may be available in isolated or pure forms; in some embodiments, drugs may be available in crude forms. In some embodiments, potential drugs may be provided as aggregates or collections, which can be screened, for example, to identify or characterize the active drug among them. In some cases, the term “drug” may refer to a compound or entity that is a polymer or contains a polymer; in some cases, the term may refer to a compound or entity that contains one or more polymer moieties. In some embodiments, the term “drug” may refer to a compound or entity that is not a polymer and / or substantially does not contain any polymer and / or one or more specific polymer moieties. In some embodiments, the term may refer to a compound or entity that lacks or substantially does not contain any polymer moieties. In some embodiments, the term may refer to a molecular complex.

[0021] Antibody: As used herein, the term “antibody” refers to a polypeptide containing standard immunoglobulin sequence elements sufficient to confer specific binding to a particular target antigen. As known in the art, intact antibodies, such as those produced naturally, are approximately 150 kDa tetramers comprising two identical heavy-chain polypeptides (each about 50 kDa) and two identical light-chain polypeptides (each about 25 kDa) that associate with each other to form a structure commonly referred to as a “Y-type” structure. Each heavy chain comprises at least four domains (each about 110 amino acids long) – an amino-terminal variable (VH) domain (located at the tip of the Y structure), followed by three constant domains: CH1, CH2, and carboxy-terminal CH3 (located at the base of the stem of Y'). A short region known as the “switch” connects the heavy-chain variable region to the constant region. The “hinge” connects the CH2 and CH3 domains to the rest of the antibody. Two disulfide bonds in this hinge region bind the two heavy-chain polypeptides together in an intact antibody. Each light chain comprises two domains, namely an amino-terminal variable (VL) domain followed by a carboxy-terminal constant (CL) domain, which are separated from each other by another “switch.” An intact antibody tetramer comprises two heavy-light chain dimers, in which the heavy and light chains are linked to each other by a single disulfide bond; two other disulfide bonds connect the heavy chain hinge regions to each other, thereby linking the dimers together and forming a tetramer. Naturally occurring antibodies are also typically glycosylated on the CH2 domain. Each domain in a natural antibody has a structure characterized by an “immunoglobulin fold” formed from two beta sheets (e.g., 3, 4, or 5-strand sheets) packed relative to each other in a compressed antiparallel beta barrel. Each variable domain contains three hypervariable loops known as “complementarity-determining regions” (CDR1, CDR2, and CDR3), and four somewhat immutable “framework” regions (FR1, FR2, FR3, and FR4).When a naturally occurring antibody folds, the FR region forms a beta sheet that provides the structural framework of the domain, and the CDR loop regions from both the heavy and light chains come together in three-dimensional space, thereby creating a single hypervariable antigen-binding site located at the tip of the Y structure. The Fc region of naturally occurring antibodies binds to elements of the complementarity system and also to receptors on effector cells, including, for example, effector cells that mediate cytotoxicity. As is known in the art, the affinity and / or other binding properties of the Fc region to Fc receptors can be modulated by glycosylation or other modifications. In some embodiments, antibodies produced and / or utilized according to the present invention include a glycosylated Fc domain, which includes a modified or altered Fc domain having such glycosylation. For the purposes of the present invention, in some embodiments, any polypeptide or polypeptide complex containing sufficient immunoglobulin domain sequences as found in natural antibodies may be referred to as an “antibody” and / or used as an “antibody,” regardless of whether such polypeptides are naturally occurring (e.g., produced by organisms that react to an antigen) or produced by recombinant technology, chemical synthesis, or other artificial systems or methodologies. In some embodiments, the antibody is polyclonal; in some embodiments, the antibody is monoclonal. In some embodiments, the antibody has a constant region sequence characteristic of mouse, rabbit, primate, or human antibodies. In some embodiments, the antibody sequence elements are humanized, primated, chimeric, etc., as known in the art. Furthermore, as used herein, the term “antibody” may, in appropriate embodiments (unless otherwise stated or evident from the context), refer to any construct or form known or developed in the art for utilizing the structural and functional features of an antibody in an alternative presentation.For example, in the embodiments, the antibodies used according to the present invention include intact IgG, IgE and IgM, bi- or multispecific antibodies (e.g., Zybody®), single-chain Fvs, polypeptide-Fc fusions, Fab, camel antibodies, masked antibodies (e.g., Probody®), and small modular immunotherapies. The antibodies are selected from, but are not limited to, ImmunoPharmaceuticals ("SMIP" trademark), single-stranded or tandem diabodies (TandAb®), VHH, Anticalin®, Nanobody®, Minibody, BiTE®, Ankyrin repeat protein or DARPIN®, Avimer®, DART, TCR-like antibodies, Adnectin®, Affilin®, Trans-body®, Affibody®, TrimerX®, MicroProteins, Fynomer®, Centyrin®, and KALBITOR®. In some embodiments, the antibodies may lack covalent modifications (e.g., glycan linkages) that they would have if naturally occurring. In some embodiments, the antibody may contain covalent modifications (e.g., the attachment of a glycan, a payload [e.g., a detectable portion, a therapeutic portion, a catalytic portion, etc.], or other pendant groups [e.g., polyethylene glycol, etc.]).

[0022] Antibody Drugs: As used herein, the term “antibody drug” refers to a drug that specifically binds to a particular antigen. In some embodiments, the term encompasses any polypeptide or polypeptide complex containing sufficient immunoglobulin structural elements to confer specific binding. Exemplary antibody drugs include, but are not limited to, human antibodies, primate-like antibodies, chimeric antibodies, bispecific antibodies, humanized antibodies, complexed antibodies (i.e., antibodies complexed or fused to other proteins, radiolabeled, or cytotoxins), small module immunotherapies ("SMIP®"), single-chain antibodies, camel antibodies, and antibody fragments. As used herein, the term “antibody drug” also includes intact monoclonal antibodies, polyclonal antibodies, single-domain antibodies (e.g., shark single-domain antibodies (e.g., IgNAR or fragments thereof)), multispecific antibodies formed from at least two intact antibodies (e.g., bispecific antibodies), and antibody fragments (only if they exhibit the desired biological activity). In some embodiments, the term encompasses staple peptides. In some embodiments, the term encompasses one or more antibody-like conjugated peptide mimetic drugs. In some embodiments, the term encompasses one or more antibody-like binding backbone proteins. In some embodiments, the term encompasses monobodies or adnectins. In many embodiments, the antibody drug is a polypeptide whose amino acid sequence includes one or more structural elements recognized by those skilled in the art as complementarity-determining regions (CDRs), or includes such a polypeptide; in some embodiments, the antibody drug is a polypeptide whose amino acid sequence includes at least one CDR (e.g., at least one heavy-chain CDR and / or at least one light-chain CDR) substantially identical to that found in a reference antibody, or includes such a polypeptide. In some embodiments, the included CDR is either identical in sequence to a reference CDR, or substantially identical to a reference CDR in that it contains 1 to 5 amino acid substitutions.In some embodiments, the included CDR is substantially identical to the reference CDR in that it exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR in that it exhibits at least 96%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments, the included CDR has at least one amino acid deleted, added, or substituted compared to the reference CDR, but otherwise the included CDR has the same amino acid sequence as the reference CDR. In some embodiments, the included CDR has 1 to 5 amino acids deleted, added, or substituted compared to the reference CDR, but otherwise is substantially identical to the reference CDR in that it has the same amino acid sequence as the reference CDR. In some embodiments, the included CDR has at least one amino acid substituted compared to the reference CDR, but otherwise is substantially identical to the reference CDR in that it has the same amino acid sequence as the reference CDR. In some embodiments, the included CDR has 1 to 5 amino acids deleted, added, or substituted compared to the reference CDR, but otherwise is substantially identical to the reference CDR in that it has the same amino acid sequence as the reference CDR. In some embodiments, the antibody drug is a polypeptide containing, or comprising, a structural element whose amino acid sequence is recognized by those skilled in the art as an immunoglobulin variable domain. In some embodiments, the antibody drug is a polypeptide protein having a binding domain that is homologous or largely homologous to an immunoglobulin binding domain. In some embodiments, the antibody drug is an antibody-drug conjugate, or comprises, one.

[0023] Antibody component: As used herein, an antibody component refers to a polypeptide element (which may be a complete polypeptide or a part of a larger polypeptide, such as a fusion polypeptide as described herein) that specifically binds to an epitope or antigen and contains structural features of one or more immunoglobulins. Generally, an antibody component is any polypeptide whose amino acid sequence contains elements characteristic of an antibody-binding region (e.g., the light chain or variable region of an antibody or one or more complementarity-determining regions ("CDRs") therein, or the heavy chain or variable region of an antibody or one or more CDRs thereof, in the presence of optionally one or more framework regions). In some embodiments, the antibody component is a full-length antibody or contains one. In some embodiments, the antibody component is less than full length but contains at least one binding site (containing at least one, preferably at least two sequences having the structure of a "variable region" of a known antibody). In some embodiments, the term "antibody component" encompasses any protein having a binding domain that is homologous or largely homologous to an immunoglobulin-binding domain. In certain embodiments, the included "antibody component" encompasses a polypeptide having a binding domain that exhibits at least 99% identity with an immunoglobulin-binding domain. In some embodiments, the “antibody component” included is any polypeptide having an immunoglobulin-binding domain, e.g., a binding domain exhibiting at least 70%, 75%, 80%, 85%, 90%, 95%, or 98% identity with a reference immunoglobulin-binding domain. The “antibody component” included may have the same amino acid sequence as that of an antibody (or a portion thereof, e.g., its antigen-binding portion) found in natural sources. The antibody component may be monospecific, bispecific, or multispecific. The antibody component may contain structural elements characteristic of any immunoglobulin class, including any of the human classes: IgG, IgM, IgA, IgD, and IgE. It has been shown that the antigen-binding function of the antibody may be performed by a fragment of a full-length antibody. Embodiments of such antibodies may also be in bispecific, dual specific, or multispecific forms that specifically bind to two or more different antigens.Examples of binding fragments included in the term "antigen-binding portion" of an antibody include: (i) a Fab fragment which is a monovalent fragment consisting of V H , V L , C H 1 and C L domains; (ii) an F(ab')2 fragment which is a bivalent fragment comprising two Fab fragments linked by disulfide bridges in the hinge region; (iii) an Fd fragment consisting of V H and C H 1 domains; (iv) an Fv fragment consisting of V H and V L domains of a single arm of an antibody; (v) a dAb fragment comprising a single variable domain (Ward et al., (1989) Nature 341:544-546); and (vi) isolated complementarity determining regions (CDRs). Further, the two domains of the Fv fragment, V H and V L are encoded by separate genes, but they can be linked by a synthetic linker which enables them to be made as a single protein chain (where the V H and V L regions pair to form a monovalent molecule, known as a single-chain Fv (scFv)); see, for example, Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). In some embodiments, an "antibody component" is, or includes, such a single-chain antibody as described herein. In some embodiments, an "antibody component" is, or includes, a diabody. A diabody is V H and V LThe domains are expressed on a single polypeptide chain, but the linker is too short to allow pairing between two domains on the same chain, thereby pairing those domains with complementary domains on another chain to create two antigen-binding sites in a bivalent, bispecific antibody (see, for example, Holliger, P., et al., (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, RJ, (1994) Structure 2(12):1121-1123). Such antibody-binding moieties are known in the art (Kontermann and Dubel eds., Antibody Engineering (2001) Springer-Verlag. New York. 790pp. (ISBN 3-540-41354-5)). In some embodiments, the antibody component consists of a pair of tandem Fv segments (V) that form a pair of antigen-binding regions together with a complementary light chain polypeptide. H -C H 1-V H -C H 1) is a single-chain "linear antibody" containing or including such an antibody (Zapata et al., (1995) Protein Eng. 8(10):1057-1062; and U.S. Patent No. 5,641,870). In some embodiments, the antibody component may have structural elements characteristic of chimeric or humanized antibodies. Generally, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues derived from the recipient's complementarity-determining region (CDR) are replaced with residues derived from the CDR of a non-human species (donor antibody), such as mouse, rat, or rabbit, having the desired specificity, affinity, and ability. In some embodiments, the antibody component may have structural elements characteristic of human antibodies.

[0024] Antibody fragment: As used herein, “antibody fragment” includes, for example, a portion of an intact antibody, such as the antigen-binding or variable region of an antibody. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; triabodies; tetrabodies; linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. For example, an antibody fragment may include an isolated fragment, an “Fv” fragment consisting of heavy and light chain variable regions, a recombinant single-chain polypeptide molecule (“ScFv protein”) in which the light and heavy chain variable regions are linked by a peptide linker, and a minimal recognition unit consisting of amino acid residues that mimic a hypervariable region. In many embodiments, the antibody fragment contains a full sequence of a parent antibody and is a fragment that binds to the same antigen as the parent antibody; in some embodiments, the fragment binds to the antigen with comparable affinity to that of the parent antibody and / or competes with the parent antibody for binding to the antigen. Examples of antibody antigen-binding fragments include, but are not limited to, Fab fragments, Fab' fragments, F(ab')2 fragments, scFv fragments, Fv fragments, dsFv diabodies, dAb fragments, Fd' fragments, Fd fragments, and isolated complementarity-determining regions (CDR) regions. Antibody antigen-binding fragments can be generated by any means. For example, antibody antigen-binding fragments may be generated enzymatically or chemically by fragmentation of an intact antibody, and / or it may be generated recombinantly from a gene encoding a partial antibody sequence. Alternatively or additionally, antibody antigen-binding fragments may be generated by whole or partial synthesis. Antibody antigen-binding fragments may optionally contain single-chain antibody fragments. Alternatively or additionally, antibody antigen-binding fragments may contain, for example, multiple chains primarily linked by disulfide bonds. Antibody antigen-binding fragments may optionally contain multimolecular complexes. Functional antibody fragments typically contain at least about 50 amino acids, and more typically at least about 200 amino acids.

[0025] Approximately: As used herein, the terms “approximately” or “about” refer to a value similar to the described reference value when applied to one or more values. In some embodiments, unless otherwise stated or evident from the context, the terms “approximately” or “about” refer to a range of values ​​that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less than or equal to the described reference value (for example, if one or more values ​​define a sufficiently narrow range, its application of such percentage discrepancies would negate the described range).

[0026] Related to: Where the term is used herein, two events or entities are “related” to each other if the presence, level and / or form of one correlates with that of the other. For example, if the presence, level and / or form of a particular entity (e.g., polypeptide, gene signature, metabolite, microorganism, etc.) correlates with the incidence and / or susceptibility to a particular disease, disorder, or condition (e.g., across a relevant population), then that entity is considered related to that disease, disorder, or condition. In some embodiments, two or more entities are physically “related” to each other if they interact directly or indirectly such that they are physically close to and / or remain close to each other. In some embodiments, two or more entities that are physically related to each other are covalently bonded to each other; in some embodiments, two or more entities that are physically related to each other are not covalently bonded to each other but are non-covalently related by, for example, hydrogen bonding, van der Waals interactions, hydrophobic interactions, magnetism, and combinations thereof.

[0027] Biocompatibility: The term “biocompatible,” as used herein, refers to a material that, for example, does not cause significant harm to living tissue when placed in contact with such tissue in vivo. In some embodiments, materials are “biocompatible” if they are not toxic to cells. In some embodiments, materials are “biocompatible” if their addition to cells in vitro results in no more than 20% cell death and / or their administration in vivo does not induce significant inflammation or other such adverse effects.

[0028] Biodegradability: As used herein, the term “biodegradable” refers to a material that, when introduced into a cell, is broken down (by cellular mechanisms such as enzymatic degradation, hydrolysis, and / or a combination thereof) into components that the cell can reuse or discard without significant toxicity to the cell. In some embodiments, the components produced by the degradation of a biodegradable material are biocompatible and therefore do not induce significant inflammation and / or other adverse effects in vivo. In some embodiments, a biodegradable polymer material degrades into its constituent monomers. In some embodiments, the degradation of a biodegradable material (including, for example, a biodegradable polymer material) includes hydrolysis of ester bonds. Alternatively or additionally, in some embodiments, the degradation of a biodegradable material (including, for example, a biodegradable polymer material) includes cleavage of urethane bonds. Exemplary biodegradable polymers include, but are not limited to, polymers of hydroxy acids such as lactic acid and glycolic acid, including, poly(hydroxyl acid), poly(lactic acid) (PLA), poly(glycolic acid) (PGA), and poly(lactic acid-coglycolic acid) (PLGA), as well as copolymers with PEG, polyanhydrides, poly(ortho)esters, polyesters, polyurethanes, poly(butyric acid), poly(valeric acid), poly(caprolactone), poly(hydroxyalkanoate), poly(lactide-co-caprolactone), blends thereof, and copolymers thereof. For example, proteins, for example, Many naturally occurring polymers, including lubumin, collagen, gelatin, and prolamin, e.g., zein, and polysaccharides, e.g., alginates, cellulose derivatives, and polyhydroxyalkanoates, e.g., polyhydroxybutyrate blends and their copolymers, are also biodegradable. Those skilled in the art can understand or determine when such polymers have biocompatible and / or biodegradable derivatives (e.g., related to a parent polymer with substantially identical structure, differing only by the substitution or addition of certain chemical groups, as is known in the art).

[0029] Biological Activators: As used herein, the term “biological activator” refers to a drug that, when administered to a subject, e.g., a human, exerts a specific biological effect. In some embodiments, a biological activator may be a therapeutic activator, a cosmetic activator, and / or a diagnostic activator. In some embodiments, a biological activator may be or include entities or parts that would be classified as “active pharmaceutical ingredients” by the U.S. Food and Drug Administration. In some embodiments, a biological activator is a large drug. In some embodiments, a biological activator may be or include a drug whose presence correlates with a desired pharmacological and / or therapeutic, cosmetic, and / or diagnostic effect. In some embodiments, a biological activator is characterized in that its biological effect is dose-dependent (e.g., optionally increases linearly with increasing dose over at least a first concentration range). In some embodiments, a drug is not considered a “biological activator” if it merely facilitates the delivery of a different drug that actually achieves the desired effect.

[0030] Botulinum macroemulsion composition: The term “botulinum macroemulsion composition” as used herein refers to any macroemulsion composition in which at least one macroemulsion contains botulinum toxin. The botulinum toxin may be present within the macroemulsion, on the surface of the macroemulsion, and / or within the micelle membrane that defines the macroemulsion.

[0031] Botulinum Nanoemulsion Composition: The term “botulinum nanoemulsion composition” as used herein refers to any nanoemulsion composition in which at least one nanoemulsion contains botulinum toxin. The botulinum toxin may be present within the nanoemulsion, on the surface of the nanoemulsion, and / or within the micelle film defining the nanoemulsion.

[0032] Botulinum Toxin The term “botulinum toxin,” as used herein, refers to any neurotoxin produced by Clostridium botulinum. Unless otherwise indicated, the term encompasses fragments or portions (e.g., light chain and / or heavy chain) of such neurotoxin that retain appropriate activity (e.g., muscle relaxant activity). The phrase “botulinum toxin,” as used herein, encompasses botulinum toxin serotypes A, B, C, D, E, F, and G. Botulinum toxin also, as used herein, encompasses both botulinum toxin complexes (i.e., complexes of 300, 600, and 900 kDa) and purified (i.e., isolated) botulinum toxin (i.e., about 150 kDa). “Purified botulinum toxin” is defined as botulinum toxin isolated from, or substantially isolated from, other proteins containing the proteins for the botulinum toxin complex. The purified toxin may have a purity of over 95%, preferably over 99%. Those skilled in the art will understand that the present invention is not limited to any particular source of botulinum toxin. For example, botulinum toxin for use according to the present invention may be isolated from Clostridium botulinum, chemically synthesized, recombinantly produced (i.e., in a host cell or an organism other than Clostridium botulinum), etc. Botulinum may be genetically engineered or chemically modified to have a longer or shorter duration of action than botulinum toxin serotype A.

[0033] Carrier: As used herein, refers to a diluent, adjuvant, excipient, or vehicle administered with the composition. In some exemplary embodiments, the carrier may include sterile liquids, such as water, and oils of petroleum, animal, plant, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. In some embodiments, the carrier is or comprises one or more solid components.

[0034] Combination Therapy: As used herein, the term “combination therapy” refers to situations in which a subject is simultaneously exposed to two or more treatment regimens (e.g., two or more therapeutic agents, therapeutic agents and therapies, etc.). In some embodiments, two or more regimens may be administered concurrently; in some embodiments, such regimens may be administered sequentially (e.g., all “doses” of the first regimen are administered before any dose of the second regimen); in some embodiments, such agents are administered in a duplicate dosing regimen. In some embodiments, “administration” of combination therapy may include the administration of one or more agents and / or therapies to a subject receiving other drugs or therapies in combination. For clarity, combination therapy does not require that individual agents be administered together in a single composition (or not necessarily even simultaneously); however, in some embodiments, two or more agents, or their active portions, may be administered together in a combination composition or even in a combination compound (e.g., as part of a single chemical complex or covalent entity).

[0035] Equivalent: As used herein, the term “equivalent” means two or more drugs, entities, situations, sets of conditions, etc., which may not be identical to one another but are similar enough to allow comparison between them, so that a reasonable conclusion can be drawn based on observed differences or similarities. In some embodiments, an equivalent set of states, situations, individuals, or groups is characterized by several substantially identical features and one or a few diverse features. A person skilled in the art will understand, in context, what degree of identity is required for two or more such drugs, entities, situations, sets of conditions, etc., to be considered equivalent in any given situation. For example, a set of situations, individuals, or groups is comparable to one another if they are characterized by a sufficient number and substantially identical types of features to justify a reasonable conclusion that differences in results or phenomena obtained under or using different sets of situations, individuals, or groups are caused by or indicate variations in those features.

[0036] Composition: Those skilled in the art will understand that the term “composition,” as used herein, may be used to refer to a separate physical entity comprising one or more specific components. Generally, unless otherwise specified, a composition may be in any form, such as a gas, gel, liquid, or solid.

[0037] "Includes": Any composition or method described herein as "including" one or more named elements or steps is open-ended, meaning that the named elements or steps are essential, but other elements or steps may be added within the scope of the composition or method. To avoid redundancy, any composition or method described as "comprising" (or "which comprises") one or more named elements or steps also describes a corresponding, more limited composition or method that "consisting essentially of" (or "which consists essentially of") the same named elements or steps, meaning that the composition or method includes the named essential elements or steps and may also include additional elements or steps that do not substantially affect the fundamental novel features of the composition or method. Any composition or method described herein as "containing" or "substantially comprising" one or more named elements or processes is also understood to describe a corresponding more limited closed-end composition or method "consisting of" (or "consists of") the named elements or processes, excluding any other unnamed elements or processes. In any composition or method disclosed herein, any known or disclosed equivalent of any named essential element or process may be used in place of that element or process.

[0038] Dosage Form or Unit Dosage Form: Those skilled in the art will understand that the term “dosage form” may be used to refer to a physically distinct unit of an active agent (e.g., a therapeutic or diagnostic agent) for administration to a subject. Typically, each such unit contains a predetermined amount of the active agent. In some embodiments, such an amount is a unit dose (or its overall fraction) appropriate for a dosing regimen that has been determined to correlate with a desired or beneficial outcome (i.e., a therapeutic dosing regimen) when administered to the population in question. Those skilled in the art will understand that the total amount of a therapeutic composition or agent to be administered to a particular subject may be determined by one or more attending physicians and may include administrations of multiple dosage forms.

[0039] Dosage regimen: Those skilled in the art will understand that the term “dosage regimen” can be used to refer to a set of unit doses (typically multiple) administered individually to a subject, typically separated by time intervals. In some embodiments, a given therapeutic agent has a recommended dosage regimen, which may comprise one or more doses. In some embodiments, a dosage regimen comprises multiple doses, each separated in time from the others. In some embodiments, the individual doses are separated from each other by periods of equal length; in some embodiments, a dosage regimen comprises multiple doses and at least two distinct periods separating the individual doses. In some embodiments, all doses within a dosage regimen are of the same unit dose amount. In some embodiments, different doses within a dosage regimen are of different amounts. In some embodiments, a dosage regimen comprises a first dose in the amount of a first dose, followed by one or more additional doses in the amount of a second dose different from the amount of the first dose. In some embodiments, the drug regimen comprises a first dose in the amount of a first dose, followed by one or more additional doses in the amount of a second dose, which is the same as the amount of the first dose. In some embodiments, the drug regimen correlates with a desired or beneficial outcome when administered across a relevant population (i.e., it is a therapeutic drug regimen).

[0040] Emulsion: The term “emulsion” as used herein is consistent with the understanding in the art of “a system consisting of a liquid dispersed with or without an emulsifier in an immiscible liquid, usually in droplets larger than colloidal size.” See, for example, the definition in Medline Plus Online Medical Dictionary, Merriam-Webster (2005).

[0041] Excipients: As used herein, these refer to non-therapeutic agents that may be included in a pharmaceutical composition to provide or contribute to a desired viscosity or stabilizing effect. Suitable pharmaceutical excipients include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, and the like.

[0042] Human: In some embodiments, a human is an embryo, fetus, infant, child, teenager, adult, or elderly person.

[0043] Hydrophilicity: As used herein, the terms “hydrophilic” and / or “polar” refer to the tendency to mix with water or to dissolve readily in water.

[0044] Hydrophobic: As used herein, the terms “hydrophobic” and / or “nonpolar” mean that a material is immiscible with water, has a tendency not to combine with water, or cannot readily dissolve in water.

[0045] To improve, increase, or decrease: As used herein or in its grammatical equivalents, the terms “improve,” “increase,” or “decrease” refer to a value relative to a baseline measurement, such as a measurement in the same individual before the initiation of the treatment described herein, or to a measurement in a control individual (or more control individuals) in the absence of the treatment described herein. In some embodiments, a “control individual” is an individual suffering from the same form of disease or injury as the individual being treated.

[0046] Large Molecules: The term “large molecules” is used herein to generally describe molecules whose size exceeds about 100 kilodaltons (KDa). In some embodiments, large molecules exceed about 110 KDa, 120 KDa, 130 KDa, 140 KDa, 150 KDa, 160 KDa, 170 KDa, 180 KDa, 190 KDa, 200 KDa, 250 KDa, 300 KDa, 400 KDa, or 500 KDa. In some embodiments, large molecules are polymers or include polymeric moieties or entities. In some embodiments, large molecules are polypeptides or include them. In some embodiments, large molecules are nucleic acids or include them.

[0047] Large-cap drugs: As used herein, the term “large-cap drugs” generally refers to drugs having a molecular weight greater than approximately 100 kilodaltons (KDa). In some embodiments, large molecules are greater than approximately 110 KDa, 120 KDa, 130 KDa, 140 KDa, 150 KDa, 160 KDa, 170 KDa, 180 KDa, 190 KDa, 200 KDa, 250 KDa, 300 KDa, 400 KDa, or 500 KDa. In some embodiments, the large-cap drugs are biological agents. In some embodiments, the large-cap drugs are one or more large molecules. In some embodiments, the large-cap drugs are one or more molecular complexes. In some embodiments, the large-cap drugs are polypeptides. In some embodiments, the large-cap drugs are complexes of polypeptides. In some embodiments, the large-cap drugs are bacterial toxins (e.g., botulinum toxin) or include them. In some embodiments, the large drug is or includes an antibody drug.

[0048] Macroemulsion: As used herein, the term “macroemulsion” refers to an emulsion in which at least some droplets have a diameter in the size range of several hundred nanometers to micrometers. As will be understood by those skilled in the art, macroemulsion is characterized by droplets with a diameter greater than 300 nm. In some embodiments, the macroemulsion composition to be used according to this disclosure comprises one or more large agents or one or more biological agents. In some embodiments, the large agents contained in the macroemulsion composition may be biological agents. As will be understood by those skilled in the art, macroemulsion compositions for use according to this disclosure may be prepared by any available means, including, for example, chemical or mechanical means. In some embodiments, the droplets in the macroemulsion have a size in the range of about 301 nm to about 1000 μm. In some embodiments, the macroemulsion has droplets with a size distribution of about 301 nm to about 1000 μm. In some embodiments, the droplets in the macroemulsion have a size in the range of about 500 nm to about 5000 μm. In some embodiments, the macroemulsion has droplets with a size distribution of approximately 500 nm to approximately 5000 μm.

[0049] Microneedle: As used herein, the term “microneedle” generally refers to an elongated structure of a length, diameter, and shape suitable for penetrating the skin. In some embodiments, microneedles are prepared and configured (either by themselves or within a device) to minimize contact with nerves when inserted into the skin, while still creating an efficient pathway for drug delivery. In some embodiments, microneedles have a constant diameter along their length. In some embodiments, microneedles have a diameter that varies along their length. In some embodiments, microneedles have a diameter that tapers along their length. In some embodiments, the diameter of the microneedle is narrowest at the tip that penetrates the skin. In some embodiments, microneedles may be solid. In some embodiments, microneedles may be hollow. In some embodiments, microneedles may be tubular. In some embodiments, microneedles may be sealed at one end. In some embodiments, multiple microneedles are utilized. In some embodiments, multiple microneedles are utilized in an array configuration. In some embodiments, microneedles may have lengths ranging from about 1 μm to about 4,000 μm. In some embodiments, the microneedles may have a length of approximately 1 μm to approximately 2,000 μm. In some embodiments, the microneedles may have a length of approximately 50 μm to approximately 400 μm. In some embodiments, the microneedles may have a length of approximately 800 μm to approximately 1,500 μm.

[0050] Nanoemulsion: As used herein, the term “nanoemulsion” refers to an emulsion in which at least some droplets have a diameter in the nanometer size range. As will be understood by those skilled in the art, nanoemulsions are characterized by droplets with a diameter of 300 nm or less. In some embodiments, the nanoemulsion compositions used in accordance with this disclosure contain one or more macrophages or one or more biological activators. In some embodiments, the macrophages contained in the nanoemulsion composition may be biological activators. As will be understood by those skilled in the art, nanoemulsion compositions for use in accordance with this disclosure may be prepared by any available means, including, for example, chemical or mechanical means. In some embodiments, the droplets in the nanoemulsion have a size in the range of about 1 nm to about 300 nm. In some embodiments, the nanoemulsion has droplets with a size distribution of about 1 nm to about 300 nm.

[0051] Nanoparticles: As used herein, the term “nanoparticles” refers to solid particles having a diameter of less than 300 nm, as defined by the National Science Foundation. In some embodiments, nanoparticles have a diameter of less than 100 nm, as defined by the National Institutes of Health.

[0052] Patient: As used herein, the term “patient” means any organism to which the provided composition is administered or may be administered for, for example, experimental, diagnostic, prophylactic, cosmetic, and / or therapeutic purposes. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, the patient is human. In some embodiments, the patient has or is susceptible to one or more disorders or conditions. In some embodiments, the patient exhibits one or more symptoms of a disorder or condition. In some embodiments, the patient has been diagnosed with one or more disorders or conditions. In some embodiments, the disorder or condition is or includes the presence of cancer or one or more tumors. In some embodiments, the patient is receiving or continues to receive a prescribed therapy to diagnose and / or treat a disease, disorder, or condition.

[0053] Penetration accelerator: As used herein, the term “penetration accelerator” refers to an agent whose presence or level correlates with increased penetration of the agent of interest through the skin compared to that observed in its absence. In some embodiments, a penetration accelerator is characterized in that it breaks down and / or disintegrates skin structures. In some embodiments, the penetration accelerator is or comprises a chemical agent (e.g., a chemical substance or enzyme). For example, chemical agents that can damage, break down, and / or decompose one or more keratinized components include, for example, alcohols such as short-chain alcohols, long-chain alcohols, or polyalcohols; amines and amides such as urea, amino acids or their esters, amides, AZONE®, derivatives of AZONE®, pyrrolidone, or derivatives of pyrrolidone; terpenes and terpene derivatives; fatty acids and their esters; macrocyclic compounds; surfactants; or sulfoxides (e.g., dimethyl sulfoxide (DMSO)), decyl methyl sulfoxide, etc.); surfactants such as anionic, cationic, and nonionic surfactants; polyols; essential oils; and / or hyaluronidases. In some embodiments, penetration promoters may be irritants in that they cause inflammatory and / or allergic reactions when the agent is applied to the skin. In some embodiments, penetration promoters are not irritants. In some embodiments, the penetration enhancer may be, or include, a chemical agent that does not damage, disintegrate, or degrade skin structures, but whose presence or level still correlates with increased penetration of the target agent through the skin compared to that observed in its absence. In some embodiments, copeptides, carrier molecules, and carrier peptides may be penetration enhancers that do not damage, disintegrate, and / or degrade skin structures. In some embodiments, copeptides, carrier molecules, and carrier peptides may be penetration enhancers that do not irritate the skin. The term “penetration enhancer” does not include mechanical devices (e.g., needles, scalpels, etc.) or their equivalents (e.g., other injury treatments).Furthermore, a person skilled in the art will understand that structures such as nanoparticles or emulsions are not chemical agents, and therefore, even if their presence correlates with the promotion of skin penetration of a target agent related to the structure, they are not chemical penetration enhancers.

[0054] Pharmaceutical Composition: As used herein, the term “pharmaceutical composition” means a composition in which an active agent is formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in an amount of a unit dose appropriate for administration in a therapeutic regimen that shows a statistically significant potential to achieve a predetermined therapeutic effect when administered to the population in question. In some embodiments, the pharmaceutical composition may be specifically formulated for administration in solid or liquid form, including, for example, a sterile solution or suspension, or a sustained-release formulation as a gel, cream, ointment, or controlled-release patch or spray applied to the skin, lung, or oral cavity; for example, as a pessary, cream, or foam for vaginal or rectal; sublingual; intraocular; transdermal; or transnasal, intrapulmonary, and other mucosal surface administration.

[0055] pharmaceutically acceptable: As used herein, the term “pharmaceutically acceptable” as applied to carriers, diluents, or excipients used to formulate the compositions disclosed herein means that the carriers, diluents, or excipients must be compatible with the other components of the composition and must not be harmful to the person taking them.

[0056] Medicinally Acceptable Carrier: As used herein, the term “medically acceptable carrier” means a medically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulant, that is involved in transporting or delivering the compound of interest from one organ or part of the body to another organ or part of the body. Each carrier must be “acceptable” in the sense that it is compatible with the other components of the formulation and is not harmful to the subject or patient. Some examples of materials that can function as pharmaceutically acceptable carriers include sugars, e.g., lactose, glucose, and sucrose; starches, e.g., corn starch and potato starch; cellulose and its derivatives, e.g., sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, e.g., cocoa butter and suppository wax; oils, e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, medium-chain triglycerides, and soybean oil; glycols, e.g., propylene glycol; polyols, e.g., glycerin, sorbitol, mannitol, and polyethylene glycol; esters, e.g., ethyl oleate and ethyl laurate; agar; buffers, e.g., magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; pH buffer solutions; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic, suitable substances used in pharmaceutical formulations.

[0057] Premix: As used herein, the term “premix” refers to any combination of components that will be used later to produce a nanoemulsion composition according to the present invention. For example, a premix is ​​any collection of components that, when subjected to high shear force, produce a nanoemulsion according to the present invention. In some embodiments, a premix is ​​a collection of components that, when subjected to high shear force, produce a nanoemulsion composition such as a homogeneous nanoemulsion composition. A premix often contains a liquid dispersion medium and other components sufficient to produce a nanoemulsion within the dispersion medium. According to some embodiments of this disclosure, one or more large agents may be included in the premix. According to some embodiments of this disclosure, one or more biological agents may be included in the premix. According to the present invention, botulinum toxin may be included in the premix. According to the present invention, one or more antibodies may be included in the premix. In some embodiments, a premix may contain one or more surfactants, penetration promoters, and / or other agents. In some embodiments, the premix comprises a solution. In some embodiments, the premix comprises botulinum toxin, an antibody, another biological agent and / or an penetration promoter, and the botulinum toxin, antibody, another biological agent and / or penetration promoter are in solution before a high shear force is applied to the premix.

[0058] Preventing or preventing: As used herein, when used in relation to the development of a disease, disorder, and / or condition, means reducing the risk of developing a disease, disorder, and / or condition, and / or delaying the onset of one or more characteristics or symptoms of a disease, disorder, or condition. Prevention may be considered complete if the onset of the disease, disorder, or condition is delayed for a predetermined period of time.

[0059] Protein: As used herein, the term “protein” means polypeptide (i.e., a chain of at least two amino acids linked together by peptide bonds). Proteins may also contain non-amino acid portions (e.g., glycoproteins, proteoglycans, etc.) and / or may be otherwise processed or modified. Those skilled in the art will understand that a “protein” may be a complete polypeptide chain (with or without a signal sequence) or a characteristic portion thereof, as produced by a cell. Those skilled in the art will understand that a protein may sometimes contain multiple polypeptide chains linked, for example, by one or more disulfide bonds or associated by other means. Polypeptides may contain L-amino acids, D-amino acids, or both, and may contain any of the various amino acid modifications or analogs known in the art. Useful modifications include, for example, terminal acetylation, amidation, methylation, etc. In some embodiments, proteins may contain native amino acids, non-native amino acids, synthetic amino acids, and combinations thereof. The term “peptide” is generally used to refer to polypeptides having a length of less than about 100 amino acids, less than about 50 amino acids, less than 20 amino acids, or less than 10 amino acids. In some embodiments, the protein is an antibody, an antibody fragment, their biologically active portion, and / or their characteristic portion.

[0060] Polypeptide: When used herein, the term “polypeptide” generally has its recognized meaning in the art of a polymer of at least three amino acids. Those skilled in the art will understand that the term “polypeptide” is intended to be sufficiently general to encompass not only polypeptides having the complete sequences listed herein, but also polypeptides representing functional fragments (i.e., fragments retaining at least one activity) of such complete polypeptides. Furthermore, those skilled in the art will understand that protein sequences generally tolerate some substitutions without disrupting activity. Therefore, any polypeptide that retains activity and shares at least about 30–40% of overall sequence identity, which is often more than about 50%, 60%, 70%, or 80%, and further usually includes at least one region of much higher identity (often more than 90% or even more than 95%, 96%, 97%, 98%, or 99%) in one or more highly conserved regions that usually contain at least 3–4 and often up to 20 or more amino acids, together with other polypeptides of the same classification. Polypeptides may contain L-amino acids, D-amino acids, or both, and may contain any of the various amino acid modifications or analogs known in the art. Useful modifications include, for example, terminal acetylation, amidation, and methylation. In some embodiments, proteins may contain native amino acids, unnatural amino acids, synthetic amino acids, and combinations thereof. The term “peptide” is generally used to refer to polypeptides having a length of less than about 100 amino acids, less than about 50 amino acids, less than 20 amino acids, or less than 10 amino acids. In some embodiments, proteins may be antibodies, antibody fragments, their biologically active portions, and / or characteristic portions thereof.

[0061] Reference: Where used herein, this refers to a standard or control on which a comparison is made. For example, in some embodiments, the drug, animal, individual, population, sample, regimen, sequence, or value under consideration is compared to a reference or control drug, animal, individual, population, sample, regimen, sequence, or value. In some embodiments, the reference or control is tested and / or determined substantially simultaneously with the test or determination under consideration. In some embodiments, the reference or control is a historical reference or control optionally embodied in a tangible medium of expression. Typically, as will be understood by those skilled in the art, the reference or control is determined or characterized under conditions or circumstances equivalent to those under evaluation. Those skilled in the art will understand that there may be similarities sufficient to justify reliance on and / or comparison with a particular possible reference or control.

[0062] Self-administration: As used herein, the term "self-administration" refers to a situation in which a subject has the ability to administer a composition to themselves without requiring medical supervision. In some embodiments of the present invention, self-administration may be carried out outside of a clinical setting. For example, in some embodiments of the present invention, a facial beauty cream may be administered by the subject at their own home.

[0063] Small molecules: Generally, in the art, “small molecules” are understood to be organic molecules with a size of less than about 5 kilodaltons (Kd). In some embodiments, small molecules are less than about 3 Kd, 2 Kd, or 1 Kd. In some embodiments, small molecules are less than about 800 daltons (D), 600 D, 500 D, 400 D, 300 D, 200 D, or 100 D. In some embodiments, small molecules are nonpolymeric. In some embodiments, small molecules are not proteins, peptides, or amino acids. In some embodiments, small molecules are not nucleic acids or nucleotides. In some embodiments, small molecules are not saccharides or polysaccharides.

[0064] Subject: As used herein, “Subject” means an organism, typically a mammal (e.g., a human, and in some embodiments, a prenatal human form). In some embodiments, the subject is suffering from the relevant disease, disorder, or condition. In some embodiments, the subject is susceptible to the disease, disorder, or condition. In some embodiments, the subject exhibits one or more symptoms or features of the disease, disorder, or condition. In some embodiments, the subject does not exhibit any symptoms or features of the disease, disorder, or condition. In some embodiments, the subject is a person who has one or more characteristics characteristic of susceptibility to or risk of the disease, disorder, or condition. In some embodiments, the subject is a patient. In some embodiments, the subject is an individual who is being diagnosed and / or has been treated.

[0065] Substantially: As used herein, the term “substantially” refers to a qualitative state indicating the overall or nearly overall degree or extent of the feature or characteristic in question. Those skilled in the art of biology will understand that biological and chemical phenomena, if any, rarely progress to completion and / or do not progress to perfection or achieve or avoid absolute results. Therefore, the term “substantially” is used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

[0066] Therapeutic Agent: As used herein, the phrase “therapeutic agent” generally refers to any agent that, when administered to an organism, elicits a desired pharmacological effect. In some embodiments, an agent is considered a therapeutic agent if it demonstrates a statistically significant effect across a suitable population. In some embodiments, a suitable population may be a population of model organisms. In some embodiments, a suitable population may be defined by a variety of criteria, such as a given age group, sex, genetic background, or pre-existing clinical condition. In some embodiments, a therapeutic agent is a substance that may be used to reduce, improve, alleviate, inhibit, prevent, delay the onset, reduce the severity, and / or decrease the incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition. In some embodiments, a “therapeutic agent” is an agent that has been approved or is required to be approved by a government agency before it can be marketed for administration to humans. In some embodiments, a “therapeutic agent” is an agent that requires a medical prescription for administration to humans. In some embodiments, an agent is not considered a “therapeutic agent” if it merely facilitates the delivery of different agents that actually achieve the desired effect.

[0067] Therapeutic effective dose: As used herein, means the amount that, when administered, produces the desired effect. In some embodiments, the term refers to an amount sufficient to treat a disease, disorder, and / or condition when administered to a population that is affected by or susceptible to the disease, disorder, and / or condition, according to a therapeutic drug regimen. In some embodiments, a therapeutic effective dose is one that reduces the incidence and / or severity of one or more symptoms of the disease, disorder, and / or condition, and / or delays their onset. Those skilled in the art will understand that the term “therapeutic effective dose” does not require that a successful treatment actually be achieved in a particular individual. Rather, a therapeutic effective dose may be an amount that, when administered to patients requiring such treatment, provides a particular desired pharmacological response in a significant number of subjects. In some embodiments, a reference to a therapeutic effective dose may refer to an amount measured in one or more specific tissues (e.g., tissues affected by the disease, disorder, or condition) or body fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc.). Those skilled in the art will understand that in some embodiments, a therapeutically effective amount of a particular drug or therapy may be formulated and / or administered in a single dose. In some embodiments, a therapeutically effective drug may be formulated and / or administered in multiple doses, for example, as part of a drug regimen.

[0068] Treatment regimen: When the term “treatment regimen” is used herein, it means a drug regimen in which administration across a relevant population may correlate with a desired or beneficial therapeutic outcome.

[0069] Treatment: As used herein, the term “treatment” (and “to treat” or “to treat”) refers to any implementation of therapy that partially or completely reduces, improves, alleviates, inhibits, delays the onset, reduces the severity, and / or decreases the incidence of one or more symptoms, features, and / or causes of a particular disease, disorder, and / or condition. In some embodiments, such treatment may be for subjects who do not show signs of the disease, disorder, and / or condition in question, and / or for subjects who show only initial signs of the disease, disorder, and / or condition. Alternatively or additionally, such treatment may be for subjects who show one or more established signs of the disease, disorder, and / or condition in question. In some embodiments, treatment may be for subjects who have been diagnosed as having the disease, disorder, and / or condition in question. In some embodiments, treatment may be for subjects who are known to have one or more susceptibility factors that are statistically correlated with an increased risk of developing the disease, disorder, and / or condition in question.

[0070] Uniform: When used herein in reference to nanoemulsion compositions, the term "uniform" refers to a nanoemulsion composition in which individual droplets have a specific range of droplet diameter sizes. For example, in some embodiments, a uniform nanoemulsion composition is one in which the difference between the minimum and maximum diameters does not exceed approximately 300 nm, 250 nm, 200 nm, 150 nm, 100 nm, 90 nm, 80 nm, 70 nm, 60 nm, 50 nm, or less. In some embodiments, droplets in a uniform large-agent nanoemulsion composition of the present invention (e.g., large-agent-containing droplets) have diameters smaller than approximately 300 nm, 250 nm, 200 nm, 150 nm, 130 nm, 120 nm, 115 nm, 110 nm, 100 nm, 90 nm, 80 nm, or less. In some embodiments, droplets in a uniform large-agent nanoemulsion composition of the present invention (e.g., large-agent-containing droplets) have diameters in the range of approximately 10 to approximately 300 nanometers. In some embodiments, droplets in the uniform large-sized drug nanoemulsion composition of the present invention have diameters in the range of about 10-300, 10-200, 10-150, 10-130, 10-120, 10-115, 10-110, 10-100, or 10-90 nm. In some embodiments, droplets in the large-sized drug nanoemulsion composition of the present invention (e.g., large drug-containing droplets) have an average droplet size of about 300, 250, 200, 150, 130, 120, or less than 115, 110, 100, or 90 nm. In some embodiments, the average droplet size is in the range of about 10-300, 50-250, 60-200, 65-150, or 70-130 nm. In some embodiments, the average droplet size is about 80-110 nm. In some embodiments, the average droplet size is about 90-100 nm. In some embodiments, the majority of droplets in the homogeneous nanoemulsion composition of the present invention (e.g., large drug-containing droplets) have a diameter less than a certain size or within a certain range. In some embodiments, the majority are greater than or equal to 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% of the droplets in the composition.In some embodiments of the present invention, a uniform nanoemulsion composition is achieved by microfluidization of the sample.

[0071] Detailed description of a prescribed embodiment Transdermal drug delivery In some embodiments, the present invention provides a technique for transdermal administration of large-capacity drugs (e.g., botulinum toxin, antibodies). In some embodiments, the disclosure teaches that particularly advantageous results are achieved when the microneedle treatment technique is combined with an emulsion composition. In some embodiments, the microneedle treatment technique is combined with a lotion, cream, or liquid composition that may later become or contain an emulsion composition (e.g., a macroemulsion composition and / or a nanoemulsion composition). In some embodiments, the provided technique does not utilize penetration enhancers. In some embodiments, the provided technique does not utilize chemical penetration enhancers that damage, break down, and / or degrade the skin. In some embodiments, the provided technique does not utilize chemical penetration enhancers.

[0072] Human skin consists of the dermis and epidermis. The epidermis has several tissue layers, namely the stratum corneum, stratum lucidum, stratum granulosum, stratum spinosum, and stratum basale (identified in order from the outer surface to the inner surface of the skin).

[0073] The stratum corneum generally presents the most significant obstacle to the transdermal delivery of large-capacity drugs. Typically about 10–15 μm thick, it consists of several layers of flattened keratinized cells (corneocytes). The intercellular spaces between corneocytes are filled with lipid structures that can play a crucial role in the penetration of substances through the skin (Bauerova et al., 2001, European Journal of Drug Metabolism and Pharmacokinetics, 26:85).

[0074] The remaining part of the epidermis beneath the stratum corneum is approximately 150 μm thick. The dermis is about 1-2 mm thick and is located beneath the epidermis. The dermis is supplied by various capillaries and nerve processes.

[0075] Transdermal administration remains a subject of research in attempts to provide alternative routes of administration that do not involve the undesirable consequences associated with injection and oral delivery. For example, needles often cause localized pain and potentially expose patients receiving injections to bloodborne diseases. Oral administration often suffers from insufficient bioavailability of the drug due to the extremely acidic environment of the patient's stomach.

[0076] In an attempt to overcome these drawbacks by providing non-invasive administration, efforts have been made to develop transdermal administration technologies for specific medicines. In transdermal administration, it is generally desirable to minimize damage to the patient's skin. Therefore, transdermal administration can reduce or eliminate injection-related pain, reduce the possibility of blood contamination, and improve the bioavailability of drugs when they are systemically incorporated.

[0077] Traditionally, attempts at transdermal administration have focused on the breakdown and / or degradation of the stratum corneum. Some attempts have involved the use of chemical penetration enhancers. Penetration enhancers may function to break down and / or decompose skin structures. In some embodiments, the penetration enhancer is or includes a chemical agent (e.g., a chemical or enzyme capable of breaking down and / or decomposing one or more stratum corneum components). In some embodiments, the penetration enhancer may be an irritant in that it may cause an inflammatory and / or allergic reaction when the agent is applied to the skin.

[0078] "However, the main limitation of penetration enhancers is that their effectiveness is often closely related to the occurrence of skin irritation." Alkilani, AZ, et al., "Transdermal drug delivery: Innovative pharmaceutical developments based on disruption of the barrier properties of the stratum corneum." Pharmaceutics. 7:438-470 (2015). Penetration enhancers tend to have inadequate efficacy and safety profiles. "They do not achieve the desired skin breakdown, and their ability to increase transport through the skin is low and variable." Ibid.

[0079] Some attempts have involved using mechanical devices to bypass or remove portions of the stratum corneum. Other attempts have involved using ultrasound or iontophoresis to facilitate the penetration of pharmaceuticals through the skin. In most cases, the aim remains to make pharmaceuticals, typically small molecules, available so that they can pass through the capillary bed in the dermis, where the drug can be systemically incorporated into the target to achieve a therapeutic effect. These methods are limited by the amount of energy that can be applied to the skin without causing discomfort and / or skin damage.

[0080] Transdermal delivery of large molecules While microneedle processing technology has been shown to facilitate the transdermal delivery of various small drugs such as calcein (approximately 623 kDa), desmopressin (approximately 1070 kDa), diclofenac (approximately 270 kDa), methyl nicotinate (approximately 40 kDa), bischloroethylnitrosourea (approximately 214 kDa), insulin (approximately 5.8 kDa), bovine serum albumin (approximately 66.5 kDa), and ovalbumin (approximately 45 kDa), the delivery of large drugs, especially those larger than 100 kDa, has remained problematic until this disclosure.

[0081] The transdermal delivery of large molecules is recognized as presenting significant challenges. Until this disclosure, microneedle treatment, particularly microneedle skin pretreatment, was not considered to affect or contribute to the transdermal administration of large drugs. For example, studies using solid microneedles for the delivery of four hydrophilic peptides—tetrapeptide-3 (456.6 Da); hexapeptide (498.6 Da); acetylhexapeptide-3 (889 Da); and oxytocin (1007.2 Da)—as well as L-carnitine (161.2 Da), showed that microneedle pretreatment significantly enhanced the penetration of each of these peptides, while the skin penetration of the peptides was molecular weight-dependent, decreasing as molecular weight increased. (Zhang, S., et al., “Enhanced delivery of hydrophilic peptides in vitro by transdermal microneedle pretreatment.” Acta Pharmaceutica) Sinica B.4(1):100-104(2014).

[0082] In a study comparing the effects of larger FITC (fluorescein isothiocyanate)-binding molecule size on transdermal delivery using sandpaper abrasion, tape stripping, and a single-puncture subcutaneous injection needle model of MSCs, it was found that for all methods, transdermal drug delivery decreased as the size of the test molecule increased (FITC complexes of 4.3, 9.6, and 42.0 kDa), similar to when tested on untreated skin. Tape stripping was the most effective technique, while sandpaper abrasion was found to cause the most skin damage. (Wu, X., et al., “Effects of pretreatment of needle puncture and sandpaper abrasion on the in vitro skin permeation of fluorescein isothiocyanate (FITC)-dextran.” International Journal of Pharmaceutics. 316:102-108 (2006)).

[0083] Other studies have attempted to deliver even larger molecules: Cascade Blue (CB, Mw538), dextran-Cascade Blue (DCB, Mw10kDa), and FITC-bound dextran (FITC-Dex, Mw72kDa). These studies used microneedles of varying lengths (300, 550, 700, or 900 μm) to puncture dermatomed human skin and evaluate the diffusion of each of the aforementioned compounds. Transport of each compound was observed with all but the 300 μm microneedle array, although degradation of DCB and FITC-Dex was observed.

[0084] As prior art demonstrates, as molecular size increases, transdermal penetration using MSCs ("microneedle skin conditioning") decreases to a negligible degree, and even to the point of being nonexistent. Even in those cases where some negligible penetration was observed, larger molecules were observed to be degraded and become biologically inactive. Therefore, until the present invention, those skilled in the art would have predicted that even with the use of mechanical or chemical penetration enhancers, large drugs such as botulinum toxin, which is approximately 150 kDa and more than twice the size of FITC-Dex, would have no penetration at all, let alone bioavailability of an intact active drug. Botulinum toxin is a complex protein, and for it to be biologically active, it requires three regions or functional parts to be intact. Therefore, damage to any one of the three regions of the protein renders it biologically inactive. According to Johnson, E et al., "Botulinum toxin is highly susceptible to denaturation due to surface denaturation, heat, and alkaline conditions." (U.S. Patent Publication No. 5512547). Therefore, significant levels of degradation and inactivation of botulinum would be expected under the microneedle treatment conditions described by Wu.

[0085] In particular, this disclosure demonstrates that microneedle treatment techniques can facilitate transdermal delivery (e.g., large drugs from macroemulsion or nanoemulsion compositions) when other penetration accelerators, especially disintegrants, are not used (i.e., no chemical penetration accelerators are used, and no other techniques that disintegrate or puncture skin structures are used).

[0086] Ultra-fine needle treatment This disclosure provides the remarkable finding that MSCs can dramatically improve the transdermal delivery of large-cap drugs. In some embodiments, the large-cap drugs may be formulated as creams and / or lotions. In some embodiments, the large-cap drugs may be combined with one or more biological agents. In some embodiments, the large-cap drugs may be formulated as or within emulsion compositions (e.g., as macroemulsions or nanoemulsions). In some embodiments, emulsions containing one or more large-cap drugs may be formulated as creams and / or lotions.

[0087] In some embodiments, microneedle (MN) arrays for use according to this disclosure are minimally invasive systems, or share similar characteristics, developed to overcome some of the drawbacks commonly associated with the use of hypodermic and subcutaneous injection needles and to improve patient comfort and compliance. Such drawbacks include, for example, the potential for mispositioning of the needle tip with subcutaneous injection needles because healthcare professionals cannot visualize exactly where the needle is going; such mispositioning can cause adverse reactions such as drooping eyelids ("ptosis") if botulinum toxin is inaccurately injected into the face. MNs would be less likely to cause such problems. Other advantages of MNs include the fact that they do not cause bleeding, minimize the introduction of pathogens through holes created by MNs, and can eliminate transdermal delivery variability. Other advantages include the possibility of self-administration, reduced risk of accidental needle stick injury, reduced risk of infection transmission, and ease of disposal. In some embodiments, the MN is a plurality of micro-protrusions assembled on one side of a support such as a patch or device (e.g., a stamp, roller, array, applicator, pen).

[0088] In some embodiments, MNs for use according to this disclosure may be designed and / or configured in arrays to improve skin contact and facilitate penetration into the skin. In some embodiments, the MNs utilized have a length, width, and shape suitable for minimizing contact with nerves when inserted into the skin, while still creating an efficient route for drug delivery. Alkilani, AZ, et al., “Transdermal drug delivery: Innovative pharmaceutical developments based on disruption of the barrier properties of the stratum corneum.” Pharmaceutics. 7:438-470 (2015).

[0089] In some embodiments, preferred MNs may be solid, coated, porous, soluble, hollow, or hydrogel MNs. Solid MNs create micropores in the skin, thereby increasing the transport of drug formulations (e.g., a "prick-and-stick" method). Coated MNs allow for rapid dissolution of coated drugs into the skin (e.g., a "coated-and-prick" method). Soluble MNs allow for rapid and / or controlled release of drugs incorporated within a microneedle. Hollow MNs can be used to puncture the skin and allow for the release of a composition after active injection or diffusion of the formulation through the microneedle puncture (e.g., a "prick-and-flow" method). In the case of soluble MNs, the MN acts as a drug depot and can hold the drug composition until released by dissolution in the case of soluble MNs, or until it swells in the case of hydrogel MNs (e.g., a "prick-and-release" method). However, as already described herein, in many embodiments, large drugs are not delivered by injection via one or more microneedles. That is, in many embodiments, any microneedle used in accordance with such embodiments is not coated, loaded, or configured with a large drug in any way that would achieve delivery of the large drug. Alternatively, in some embodiments, as described herein, the MN (whether or not it is an MSC) used in accordance with this disclosure may contain and / or deliver a large drug if the large drug is formulated in a macro or nanoemulsion composition as described herein. Therefore, as will be understood by those skilled in the art reading the specification described herein, skin treatment with a microneedle(s) that delivers a large drug (e.g., by injection through the microneedle, by release of the microneedle coating, or by release from a dissolved microneedle) is not microneedle skin conditioning.

[0090] In some embodiments, the microneedle has a constant diameter along its entire length. In some embodiments, the diameter of the microneedle is greatest at its proximal end. In some embodiments, the microneedle tapers to a point distal to its base. In some embodiments, the microneedle may be solid. In some embodiments, the microneedle may be hollow. In some embodiments, the microneedle may be tubular. In some embodiments, the microneedle may be sealed at one end. In some embodiments, the microneedle is part of an array of microneedles. In some embodiments, the microneedle may have a length of about 1 μm to about 4,000 μm. In some embodiments, the microneedle may have a length of about 1 μm to about 2,000 μm. In some embodiments, the microneedle may have a length of about 50 μm to about 400 μm. In some embodiments, the microneedle may have a length of about 800 μm to about 1,500 μm.

[0091] In some embodiments, microneedles for use according to this disclosure may be composed of various materials using techniques including, but not limited to, micromolding processes or lasers. In some embodiments, MNs may be manufactured using various types of biocompatible materials, including polymers, metals, ceramics, semiconductors, organic materials, composites, or silicon. Unless they are designed to cleave and dissolve in the skin, in some embodiments, microneedles have the mechanical strength to deliver drugs or collect biological fluids while remaining intact during insertion into and / or removal from the skin after insertion. In some embodiments, MNs can remain in place for up to several days before intact removal. In some embodiments, microneedles may be sterilizable using standard techniques. In some embodiments, MNs are biodegradable. In some embodiments, MNs include polymer materials. In some embodiments, the polymer material includes poly-L-lactic acid, polyglycolic acid, polycarbonate, polylactic acid-coglycolic acid (PLGA), polydimethylsiloxane, polyvinylpyrrolidone (PVP), copolymer of methyl vinyl ether and maleic anhydride, sodium hyaluronate, carboxymethylcellulose, maltose, dextrin, galactose, starch, gelatin, or a combination thereof.

[0092] MN arrays and MSC devices suitable for use in combination with compositions containing large-capacity drugs for transdermal delivery of large-capacity drugs include, for example, devices described in U.S. Patents 6,334,856; 6,503,231; 6,908,453; 8,257,324; and 9,144,671.

[0093] Large-scale drug In some embodiments, the compositions provided and / or utilized as described herein comprise one or more macrophages. In some embodiments, the macrophages utilized are biological activators (e.g., therapeutic activators). In particular, this disclosure provides methods for topical administration of compositions comprising macrophages in combination with MSCs.

[0094] 1. Protein preparations A variety of protein preparations may be incorporated into the provided composition and administered in combination with MSCs. In some embodiments, the protein preparation may be a peptide preparation. In some embodiments, the peptide has a molecular weight greater than 100 kDa. In some embodiments, the peptide preparation has a molecular weight of at least 150 kDa. In some embodiments, the peptide preparation consists only of naturally occurring amino acids. In some embodiments, the peptide preparation contains one or more amino acids that do not exist naturally.

[0095] (i) Botulinum toxin In some embodiments, the major agent may be botulinum toxin. Botulinum toxin (BTX) is a potent polypeptide neurotoxin produced naturally by the anaerobic Gram-positive bacillus Clostridium botulinum. Most notably, BTX causes a neuroparalytic disease in humans and animals known as botulism. BTX can apparently cross the lining of the gastrointestinal tract and attack peripheral motor neurons. Symptoms of botulism can progress from difficulty walking, swallowing, and speaking to respiratory paralysis and death.

[0096] The molecular weight of the botulinum toxin protein molecule for all seven known botulinum toxin serotypes is approximately 150 kDa. Botulinum toxin is released by Clostridium bacteria as a complex containing a 150 kDa botulinum toxin protein molecule along with associated non-toxic proteins. Therefore, the BTX-A complex can be produced by Clostridium bacteria in 900 kDa, 500 kDa, and 360 kDa forms. Botulinum toxins B and C1 are apparently produced only as a 500 kDa complex. Botulinum toxin D is produced as both 300 kDa and 500 kDa complexes. Finally, botulinum toxins E and F are produced only as a complex of approximately 300 kDa.

[0097] BTX complexes (i.e., their compositions having a molecular weight greater than approximately 150 kDa) are thought to contain non-toxic hemagglutinin proteins and non-toxic, non-aggregate proteins. These two non-toxic proteins (including the neurotoxic complex associated with the botulinum toxin molecule) may act to provide stability against denaturation of the botulinum toxin molecule and protection against digestive acids when the toxin is ingested.

[0098] Either the BTX protein or the BTX complex can be used as a known therapeutic agent and / or independently as an active biological agent in accordance with the present invention. Indeed, it will be understood by those skilled in the art that any portion or fragment of the BTX protein or complex retaining appropriate activity can be used as described herein.

[0099] In some embodiments, the botulinum toxin may be selected from the group consisting of types A, Ab, Af, B, Bf, C1, C2, D, E, F, and G; their mutants; their variants; their fragments; their characteristic parts; and / or their fusions. In some embodiments, the botulinum toxin exists as one of the subtypes described in Sakaguchi, 1982, Pharmacol.Ther., 19:165; and / or Smith et al., 2005, Infect.Immun., 73:5450 (both of which are incorporated herein by reference).

[0100] In some embodiments, the present invention provides botulinum toxin compositions. In some embodiments, the present invention provides nanoemulsion botulinum toxin compositions. Commercial sources of botulinum toxin that can be used according to the present invention include, but are not limited to, BOTOX®, DYSPORT® (Clostridium botulinum type A toxin hemagglutinin complex with human serum albumin and lactose; Ispen Limited, Berkshire UK), Xeomin®, PurTox®, Medy-Tox, NT-201 (Merz Pharmaceuticals), and / or MYOBLOC® (injectable solution consisting of botulinum toxin type B, human serum albumin, sodium succinate, and sodium chloride, pH 5.6, Elan Pharmaceuticals, Dublin, Ireland), NEURONOX (Medytox), HENGLI (Lanzhou Institute), and others. Those skilled in the art will be aware of standard and / or approved dosing regimens for such commercially available botulinum toxin compositions and will understand that any relevant such composition and / or regimen may be used in conjunction with microneedle processing techniques (e.g., in particular MSC) as described herein.

[0101] In some embodiments, the provided composition, comprising a botulinum toxin composition and formulated as a cream and / or lotion, contains about 1 to about 50,000 units of botulinum toxin per mL. In some embodiments, the provided composition, comprising a botulinum toxin composition and formulated as a cream and / or lotion, contains about 500 to about 20,000 units of botulinum toxin per mL. In some embodiments, the provided composition, comprising a botulinum toxin composition and formulated as a cream and / or lotion, contains about 100 to about 2,000 units of botulinum toxin per mL. In some embodiments, the provided composition, comprising a botulinum toxin composition and formulated as a cream and / or lotion, contains about 50 to about 500 units of botulinum toxin per mL. In some embodiments, the provided composition, comprising a botulinum toxin composition and formulated as a cream and / or lotion, contains about 25 to about 400 units of botulinum toxin per mL.

[0102] In some embodiments, the botulinum toxin composition contains about 2 to about 40,000 units of botulinum toxin per mL. In some embodiments, the botulinum toxin composition contains about 2 to about 12,000 units of botulinum toxin per mL. In some embodiments, the botulinum toxin composition contains about 100 to about 2,000 units of botulinum toxin per mL. In some embodiments, the botulinum toxin composition contains about 50 to about 1,000 units of botulinum toxin per mL.

[0103] In some embodiments, the botulinum toxin composition comprises at least one biological activator other than botulinum toxin. Alternatively or additionally, in some embodiments, the botulinum composition is administered in combination with at least one other composition comprising such a biological activator. In some embodiments, the botulinum composition is administered in combination with an penetration promoter. In some embodiments, the botulinum composition is administered in combination with another biological activator. In some embodiments, the botulinum composition is administered in combination with another biological activator and an penetration promoter.

[0104] In some embodiments, the biological activators used in combination with the botulinum toxin described herein may be agents that act on or within the skin and / or impart therapeutic and / or cosmetic effects. For example, in some embodiments, such biological activators may be selected from therapeutic agents such as anesthetics (e.g., lidocaine), steroids (e.g., hydrocortisone), and / or retinoids (e.g., retin-A), and cosmetic agents such as skin fillers, collagen, and / or silicones. In some embodiments, the botulinum composition is administered in combination with a delivery modifier such as an penetration enhancer (in some embodiments, not an irritant and / or not decomposing, disintegrating, and / or damaging skin structures and / or skin).

[0105] In some embodiments, the non-irritating penetration promoter may be selected from, for example, a copolymer, a carrier molecule, and a carrier peptide. In some embodiments, the carrier molecule is positively charged. In some embodiments, the carrier molecule may be a copolymer. In some embodiments, the carrier molecule may be a long-chain positively charged polypeptide or a positively charged non-peptidyl polymer, such as a polyalkylene imine. In some embodiments, the carrier peptide may be a cationic peptide. In some embodiments, the carrier peptide has the sequence RKKRRQRRRG-(K) 15A positively charged carrier having -GRKKRRQRRR. In some embodiments, the carrier molecule may be one disclosed in U.S. Patent Publication 2010 / 0168023 or U.S. Patent Publication 2009 / 0247464, the contents of which are incorporated herein by reference in their entirety.

[0106] In some embodiments, the provided composition, comprising both a botulinum toxin nanoemulsion composition and a cream and / or lotion formulation, contains about 1 to about 50,000 units of botulinum toxin per mL. In some embodiments, the provided composition, comprising both a nanoemulsion composition and a cream and / or lotion formulation, contains about 500 to about 20,000 units of botulinum toxin per mL. In some embodiments, the provided composition, comprising both a nanoemulsion composition and a cream and / or lotion formulation, contains about 100 to about 2,000 units of botulinum toxin per mL. In some embodiments, the provided composition, comprising both a botulinum toxin nanoemulsion composition and a cream and / or lotion formulation, contains about 50 to about 500 units of botulinum toxin per mL. In some embodiments, the provided composition, comprising both a botulinum toxin nanoemulsion composition and a cream and / or lotion formulation, contains about 25 to about 400 units of botulinum toxin per mL.

[0107] In some embodiments, the botulinum toxin nanoemulsion composition contains about 2 to about 40,000 units of botulinum toxin per mL. In some embodiments, the botulinum toxin nanoemulsion composition contains about 2 to about 12,000 units of botulinum toxin per mL. In some embodiments, the botulinum toxin nanoemulsion composition contains about 100 to about 2,000 units of botulinum toxin per mL. In some embodiments, the botulinum toxin nanoemulsion composition contains about 50 to about 1,000 units of botulinum toxin per mL.

[0108] (ii) Antibody drugs In some embodiments, the disclosure relates to the delivery of antibody drugs. In some embodiments, the large drug may be an antibody or a fragment or derivative thereof. In particular, the disclosure provides certain compositions comprising antibody drugs and also provides techniques for administering compositions comprising antibody drugs, such administration being combined with MSCs.

[0109] In some embodiments, antibody drugs may be suitable for treating skin conditions. In some embodiments, antibody drugs may be fusion proteins. In some embodiments, antibody drugs may be conjugated to other parts. In some embodiments, antibody drugs may be conjugated to polyethylene glycol.

[0110] In some embodiments, the antibody drug targets TNFα (including epitope-binding elements found in anti-TNFα antibodies such as infliximab, adalimumab, golimumab, etanercept, etanercept-szzs, and / or certolizumab pegol). In some embodiments, the antibody drug targets CD2 (including epitope-binding elements found in anti-CD2 antibodies such as ciprizumab). In some embodiments, the antibody drug targets CD4 (including epitope-binding elements found in anti-CD4 antibodies such as zanorimumab).

[0111] In some embodiments, the antibody drug targets IL-12 (including epitope-binding elements found in anti-IL-12 antibodies such as briakinumab). In some embodiments, the antibody drug targets IL-17 (including epitope-binding elements found in anti-IL-17 antibodies such as secukinumab and / or brodalumab). In some embodiments, the antibody drug targets IL-22 (including epitope-binding elements found in anti-IL-22 antibodies such as fezakinumab). In some embodiments, the antibody drug targets IL-23 (including epitope-binding elements found in anti-IL-22 antibodies such as ustekinumab and / or guselkumab).

[0112] In some embodiments, the antibody drug composition comprises at least one biological agent other than the antibody drug. Alternatively or additionally, in some embodiments, the antibody drug composition is administered in combination with at least one other composition comprising such a biological agent. In some embodiments, the antibody drug composition is administered in combination with an penetration promoter. In some embodiments, the antibody drug composition is administered in combination with another biological agent. In some embodiments, the antibody drug composition is administered in combination with another biological agent and a penetration promoter. In some embodiments, the antibody drug composition is a nanoemulsion. In some embodiments, the antibody drug composition is a cream and / or lotion formulation.

[0113] In some embodiments, the biological activators used in combination with the antibody drugs described herein may be agents that act on or within the skin and / or impart therapeutic and / or cosmetic effects. For example, in some embodiments, such biological activators may be selected from therapeutic agents such as anesthetics (e.g., lidocaine), steroids (e.g., hydrocortisone), and / or retinoids (e.g., retin-A), and cosmetic agents such as skin fillers, collagen, and / or silicones. In some embodiments, the antibody drug composition is administered in combination with a delivery modifier such as an penetration enhancer (in some embodiments, not an irritant and / or not decomposing, disintegrating, and / or damaging the skin structure(s) and / or skin).

[0114] In some embodiments, the non-irritating penetration promoter may be selected from, for example, a copolymer, a carrier molecule, and a carrier peptide. In some embodiments, the carrier molecule is positively charged. In some embodiments, the carrier molecule may be a copolymer. In some embodiments, the carrier molecule may be a long-chain positively charged polypeptide or a positively charged non-peptidyl polymer, such as a polyalkylene imine. In some embodiments, the carrier peptide may be a cationic peptide. In some embodiments, the carrier peptide has the sequence RKKRRQRRRG-(K) 15A positively charged carrier having -GRKKRRQRRR. In some embodiments, the carrier molecule may be one disclosed in U.S. Patent Publication 2010 / 0168023 or U.S. Patent Publication 2009 / 0247464, the contents of which are incorporated herein by reference in their entirety.

[0115] 2. Preventive medications A variety of prophylactic agents may be incorporated into the provided composition and administered in combination with MSCs according to the present invention. In some embodiments, the prophylactic agent includes, but is not limited to, a vaccine. In some embodiments, the vaccine may include isolated proteins or peptides, inactivated organisms and viruses, dead organisms and viruses, genetically modified organisms or viruses, and cell extracts. In some embodiments, the prophylactic agent may be combined with interleukins, interferons, cytokines, and adjuvants, such as cholera toxin, alum, Freund's adjuvant, etc. In some embodiments, the prophylactic agent includes Streptococcus pnuemoniae, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyrogenes, Corynebacterium diphtheriae, Listeria monocytogenes, Bacillus anthracis, Clostridium tetani, Clostridium botulinum, Clostridium perfringens, Neisseria meningitidis, Neisseria gonorrhoeae, Streptococcus mutans, Pseudomonas aeruginosa, Salmonella typhi, Haemophilus parainfluenzae, Bordetella pertussis, Francisella tularensis, Yersinia pestis, Vibrio cholerae, Legionella pneumophila, Mycobacterium tuberculosis, Mycobacterium leprae, Treponema pallidum, Leptospirosis Antigens of bacterial organisms such as *Correlia interrogans*, *Borrelia burgdorferi*, and *Camphylobacter jejuni*;Antigens of viruses such as smallpox, influenza A and B, respiratory syncytial virus, parainfluenza, measles, HIV, varicella-zoster virus, herpes simplex virus types 1 and 2, cytomegalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps, rabies, rubella, coxsackievirus, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, hepatitis A, B, C, D, and E viruses; Cryptococcus neoformans, Histoplasma capsulatum, Candida albicans, Candida tropicalis, Nocardia asteroides, Rickettsia ricketsii, Rickettsia typhi, Mycoplasma pneumoniae, Chlamydial psittaci, Chlamydial trachomatis, Plasmodium falciparum, Trypanosoma Antigens of fungi, protozoa, and parasites such as brucei, Entamoeba histolytica, Toxoplasma gondii, Trichomonas vaginalis, and Schistosoma mansoni may be included. In some embodiments, these antigens may be in the form of total dead organisms, peptides, proteins, glycoproteins, carbohydrates, or combinations thereof.

[0116] Those skilled in the art will recognize that the preceding paragraphs provide an exemplary, non-exclusive list of agents that can be delivered using the techniques of the present invention. Any agent may be associated with a composition provided in accordance with the present invention.

[0117] Topical preparations The compositions described herein are particularly useful in that they can be used to deliver large-capacity drugs to subjects in need via topical and / or transdermal administration (e.g., by lotion, cream, powder, ointment, liniment, gel, drop, etc.). In some embodiments, the provided cream and / or lotion formulations containing a large-capacity drug are administered to subjects in need via topical and / or transdermal administration (e.g., by lotion, cream, powder, ointment, liniment, gel, drop, etc.).

[0118] In some embodiments, the cream and / or lotion formulation comprises purified water, methylparaben, mineral oil, isopropyl myristate, white petrolatum, emulsifying wax, and propylparaben.

[0119] In some embodiments, the present invention provides specific cream and / or lotion formulations as described herein. In some embodiments, the provided cream and / or lotion formulations contain water. In some embodiments, the provided cream and / or lotion formulations contain methylparaben. In some embodiments, the provided cream and / or lotion formulations contain mineral oil. In some embodiments, the provided cream and / or lotion formulations contain isopropyl myristate. In some embodiments, the provided cream and / or lotion formulations contain white petrolatum. In some embodiments, the provided cream and / or lotion formulations contain emulsifying wax. In some embodiments, the provided cream and / or lotion formulations contain propylparaben. In some embodiments, the provided cream and / or lotion formulations do not contain any parabens. In some embodiments, the provided cream and / or lotion formulations do not contain methylparaben. In some embodiments, the provided cream and / or lotion formulations do not contain propylparaben. Exemplary lotion formulations are provided in Table 1. [Table 1]

[0120] In some embodiments, the cream and / or lotion formulations may be useful for topical and / or transdermal administration. The present invention encompasses the recognition that the cream and / or lotion formulations provided may be particularly useful for the delivery of a drug to the dermis layer of the skin. In some embodiments, the cream and / or lotion formulations provided are formulated for topical and / or transdermal delivery to a target that requires it. In some embodiments, the cream and / or lotion formulations provided are administered to a target that requires it via topical and / or transdermal delivery.

[0121] In some embodiments, the provided composition is formulated using cosmetically acceptable ingredients. For example, in some embodiments, the provided composition is formulated using water and any other cosmetically acceptable solvent, in particular monoalcohols used alone or in mixtures, such as alkanols having 1 to 8 carbon atoms (e.g., ethanol, isopropanol, benzyl alcohol, and phenylethyl alcohol), polyalcohols, such as alkylene glycols (e.g., glycerin, ethylene glycol, and propylene glycol), and glycol ethers, such as mono-, di-, and tri-ethylene glycol monoalkyl ethers, such as ethylene glycol monomethyl ether and diethylene glycol monomethyl ether. Such ingredients may be present in proportions of up to 60%, 70%, 80%, or 90% by weight of the total composition.

[0122] In some embodiments, the composition provided for topical administration comprises one or more cosmetically acceptable components that impart desirable or appropriate appearance characteristics to the subject to which the composition is to be applied (e.g., a matte appearance which may be particularly desirable or appropriate for administration to a subject having oily skin).

[0123] In some embodiments, the provided composition is formulated using at least one cosmetically acceptable filler to obtain a matte product, for example, which may be particularly desirable for individuals with oily skin.

[0124] In some embodiments, large-cap drugs are formulated into compositions suitable for topical administration. Exemplary large-cap drugs include those described herein. In some embodiments, the provided compositions may be formulated and delivered in combination with MSCs so as to achieve systemic delivery; in some embodiments, the provided compositions may be formulated and / or delivered so as to achieve topical but non-systemic delivery.

[0125] In some embodiments, compositions suitable for topical formulations include an penetration enhancer. In some embodiments, the penetration enhancer breaks down, disintegrates, and / or damages skin structures and / or skin. In some embodiments, the penetration enhancer does not break down, disintegrate, and / or damage skin structures and / or skin. In some embodiments, the penetration enhancer is an irritant. In some embodiments, the penetration enhancer is not an irritant.

[0126] This disclosure specifically demonstrates the effective and efficient delivery of therapeutic agents (and, in particular, large biologics such as botulinum toxin and / or antibody drugs) to the dermis using compositions provided in combination with MSCs. For example, in some embodiments, the present invention provides a method comprising the administration of the compositions described herein without clinically significant side effects. One example of clinically significant side effects when local delivery is intended is, but is not limited to, undesirable systemic side effects, damage to nerve tissue beneath the dermis (e.g., nerve paralysis), undesirable effects on muscles (e.g., muscle paralysis), and / or undesirable blood levels of the therapeutic agent.

[0127] Those skilled in the art will understand that the provided composition can be incorporated into a device, for example, a patch. Various transdermal patch structures are known in the art; those skilled in the art will understand that the provided composition can be easily incorporated into any of various such structures. In some embodiments, the transdermal patch may include a number of needles extending from one side of the patch applied to the skin, the needles extending from the patch and protruding through the stratum corneum of the skin. In some embodiments, the needles do not rupture blood vessels. In some embodiments, the needles do not penetrate deeply enough to reach nerves in the dermis of the skin.

[0128] In some embodiments, the transdermal patch includes an adhesive. Several examples of adhesive patches are well known (see, for example, U.S. Design Patent No. 296,006; and U.S. Patents No. 6,010,715; No. 5,591,767; No. 5,008,110; No. 5,683,712; No. 5,948,433; and No. 5,965,154; all of which are incorporated herein by reference). An adhesive patch is generally characterized by having an adhesive layer applied to the patient's skin, a depot or retainer for holding the composition to be provided, and an outer surface to prevent leakage of the composition to be provided from the depot. The outer surface of the patch may be non-adhesive.

[0129] According to the present invention, the provided composition is incorporated into a patch so as to be stable over a long period of time. For example, in some embodiments, the provided composition may be incorporated into a polymer matrix that stabilizes a large drug and allows the drug to diffuse from the matrix and the patch. The provided composition may also be incorporated into an adhesive layer of the patch so that the provided composition can diffuse through the skin when the patch is applied to the skin. In some embodiments, the adhesive layer may be thermally activated when the temperature is about 37°C, which gradually liquefies the adhesive so that the drug can diffuse through the skin. The adhesive may remain sticky when stored below 37°C, and when applied to the skin, the adhesive loses its stickiness as it liquefies.

[0130] In some embodiments, the composition provided may be delivered in a depot within a patch such that pressure applied to the patch causes the composition to be delivered through a microneedle and through the stratum corneum toward the outside of the patch. Exemplary embodiments of the microneedle are described above. Apparatus suitable for use when administering the composition provided intradermally includes apparatus described in U.S. Patents 4,886,499; 5,190,521; 5,328,483; 5,527,288; 4,270,537; 5,015,235; 5,141,496; and 5,417,662. The intradermal composition may be administered by apparatus that limits the effective penetration length of the needle into the skin, such as those described in PCT Publication WO99 / 34850 and its functional equivalents.

[0131] In some embodiments, for example, it may be desirable to slow down the absorption of the composition into the skin in order to prolong the effects of the composition. In some embodiments, this can be achieved by using a liquid suspension of a crystalline or amorphous material having insufficient water solubility. The absorption rate of the composition then depends on its dissolution rate, which may also depend on the crystal size and crystalline form. In some embodiments, the release rate of the composition can be controlled depending on the ratio of the composition to the polymer and the properties of the particular polymer used. Other examples of biodegradable polymers include poly(orthoester) and poly(anhydrous).

[0132] Emulsion This disclosure includes the recognition that emulsion technology may provide stabilization benefits to target drugs, including, in particular, botulinum toxin and / or antibody drugs, as described herein.

[0133] Furthermore, this disclosure recognizes that certain liquid nanoemulsion technologies have been demonstrated to provide remarkable transdermal delivery properties even for very large molecules such as botulinum and / or antibody drugs. See, for example, U.S. Patent Publication 2012 / 0328701, U.S. Patent Publication 2012 / 0328702, No. 8,318,181, and U.S. Patent No. 8,658,391 (these disclosures are incorporated herein by reference in their entirety). These liquid nanoemulsions are far superior to solid nanoparticle drug delivery, particularly transdermal drug delivery where solid nanoparticles cannot penetrate the skin and only accumulate in hair follicles, as described by Gomaa. These liquid nanoemulsions are also stable for at least 34 months, which also makes them commercially viable.

[0134] 1. Macroemulsion In some embodiments, the disclosure provides strategies for using microneedle treatment to “modify” the skin to which a transdermal product has been, is being, or will be applied. Studies analyzing the transdermal delivery of small molecules (specifically, short hydrophilic peptides having molecular weights in the range of 400–1000 Da) have shown that “skin penetration of peptides depends on their molecular weight and decreases as molecular weight increases” (Zhang, S., et al., “Enhanced delivery of hydrophilic peptides in Despite previous reports suggesting that such strategies are likely only useful for drugs with low molecular weights, this disclosure provides the insight that such microneedle preparations can, surprisingly, offer significant benefits in facilitating the transdermal delivery of large drugs (e.g., those with molecular weights of approximately 100 kDa or greater).

[0135] This disclosure provides insight that the effective and rapid (i.e., administration within minutes) transdermal delivery of large molecules by such liquid macroemulsion compositions can, surprisingly, be improved by combining macroemulsion administration with microneedle dermal conditioning (MSC). This disclosure particularly demonstrates that microneedle preparation techniques (e.g., microneedle preparation of the skin) can significantly enhance the transdermal delivery of large-capacity drugs (e.g., botulinum toxin and / or antibody drugs), especially when used in conjunction with macroemulsion techniques. The specific macroemulsion compositions covered include water-in-oil and oil-in-water macroemulsions characterized by droplet sizes ranging from about 300 nm to about 5,000 μm in diameter, an oil-to-aqueous dispersion ratio ranging from about 0.01:1 to about 20:1, an oil-to-surfactant ratio ranging from about 0.1 to about 40, and / or a zeta potential ranging from about -80 mV to about +80 mV. The surfactant portion of the composition may contain one or more surfactants. Macroemulsion formulation [Table 6] *Those skilled in the art may, in consideration of this specification, make reasonable adjustments to this and other components depending on the volume, weight, and / or dosage of the large-capacity drug used.

[0136] Macroemulsion formulations may act to stabilize large-cap drugs such as botulinum toxin and / or antibody drugs. While macroemulsion formulations themselves may not necessarily be expected to achieve transdermal delivery of large-cap drugs, this disclosure nevertheless encompasses the insight that the improved stabilization that may be provided by their incorporation into macroemulsion compositions, when combined with the microneedle processing techniques described herein, may achieve synergistic enhancement of transdermal delivery.

[0137] This disclosure teaches that, despite the expectation that MSCs are only useful in facilitating the transdermal delivery of small compounds, the transdermal delivery of large drugs by macroemulsion compositions can be enabled in combination with microneedle technology. Furthermore, this disclosure is particularly surprising considering that microneedle preparation, combined with the encapsulation of small molecule drugs in solid nanoparticles, as described by Gomaa, provided only a small amount of penetration as early as 6 hours after administration, and no material penetration was observed until 24 hours after administration.

[0138] In some embodiments, a macroemulsion formulation containing a large drug is administered in conjunction with microneedle preparation using a solid microneedle. In some embodiments, site MSC is performed before applying the macroemulsion formulation containing the large drug to that site (e.g., before a specific application and / or before each application). In some embodiments, site MSC is performed after applying the macroemulsion formulation containing the large drug to that site. In some embodiments, site MSC and application of the macroemulsion formulation containing the large drug to that site occur substantially simultaneously. In some embodiments, the macroemulsion formulation containing the large drug is not injected via one or more microneedles. In some embodiments, the microneedle is part of an array of microneedles. In some embodiments, the microneedle may have a length of about 1 μm to about 4,000 μm. In some embodiments, the microneedle may have a length of about 1 μm to about 2,000 μm. In some embodiments, the microneedle may have a length of about 50 μm to about 400 μm. In some embodiments, the microneedles may have a length of approximately 800 μm to approximately 1500 μm.

[0139] The findings presented herein are particularly surprising, considering reports that transdermal delivery of solid nanoparticles much smaller than droplets in the macroemulsion compositions used herein (e.g., 10⁵ ± 2.92 nm) does not effectively deliver (or facilitate) even small molecule drugs transdermally through the skin. For example, Gomaa et al. al described a study in which a solution of rhodamine dye (molecular weight 479 Da) encapsulated in PLGA nanoparticles was applied to skin pre-treated by microneedle treatment to evaluate skin penetration. See Gomaa, Y., et al, “Effect of microneedle treatment on the skin permeation of a nanoencapsulated dye.” J Pharm Pharmacol. 2012 November;64(11):1592-1602. The data showed that very small amounts of dye began to penetrate the skin after 6 hours of continuous application; no significant increase in penetration was observed until the skin was treated continuously for 24 hours. The researchers explained that “there is a new consensus that NP [nanoparticles] cannot normally penetrate the stratum corneum but can accumulate well within hair follicles.” Therefore, prior to this disclosure, those skilled in the art would anticipate that the use of microneedle processing techniques using vehicles considerably larger than 105 nm would not be considered effective for transdermal delivery of even small molecule drugs (e.g., rhodamine dyes); and that delivery of large molecules would certainly be impossible. However, this disclosure demonstrates that microneedle processing can significantly facilitate transdermal delivery of large molecules, particularly when used in conjunction with macroemulsion techniques.

[0140] In particular, this disclosure demonstrates that microneedling techniques can facilitate transdermal delivery (e.g., large drugs from macroemulsion compositions) when no other disintegrants are used (i.e., no chemical penetration enhancers are used, and no other techniques are used to disintegrate or puncture skin structures). Previous studies of transdermal delivery of drugs roughly the size of botulinum toxin (i.e., about 150 kDa) using microneedling have reported that delivery is unsuccessful unless additional treatment is applied to disintegrate the skin. For example, U.S. Patent Publication 2010 / 0196445 reports that botulinum toxin is not effectively delivered from pre-coated microneedles unless cutaneous digestive enzymes are also applied to disintegrate skin structures at the microneedling site.

[0141] This disclosure demonstrates, in particular, that microneedle processing techniques can achieve transdermal delivery (e.g., of large-capacity drugs from microemulsion and nanoemulsion compositions) when microneedle coating or loading is not utilized and / or when the microneedles are not designed to remain in the skin. In particular, as already described, this disclosure understands that such microneedle coating or loading may not be commercially viable, at least due to the instability of the botulinum coating or loading material. For example, Johnson, E., et al. According to al., "Botulinum toxin is highly susceptible to denaturation due to surface denaturation, heat, and alkaline conditions. Lyophilization or freeze-drying of botulinum toxin is the most economically rational and practical method of distributing the product in a stable form that is readily available to clinicians." U.S. Patent No. 5,512,547. Also, as will be understood by those skilled in the art reading this specification, the techniques described herein have certain advantages, including the fact that it is not necessary to leave microneedles in or with the tissue. For example, those skilled in the art will understand that leaving microneedles in the skin may carry the risk of skin irritation, inflammation, allergic reactions, and / or cosmetically undesirable scarring. In contrast to the present invention, techniques such as those described in U.S. Patent Publication No. 2017 / 0209553 utilize a microneedle array designed in which botulinum toxin is loaded into the needle and the microneedle is detached within the skin (as per U.S. Patents Nos. 2017 / 0209553 and 2016 / 0263362).

[0142] This disclosure provides a remarkably effective technique for the transdermal delivery of large-cap drugs (e.g., botulinum toxin, antibodies, etc.). In particular, this disclosure teaches that the transdermal delivery of such drugs can be significantly facilitated through the use of microneedle processing techniques without any other disintegration measures. Thus, the techniques provided can achieve effective delivery without the inflammation, irritation, and / or allergic reactions that often occur with the use of skin-disintegrating agents. As will be understood by those skilled in the art reading this specification, this disclosure teaches that the transdermal delivery of large-cap drugs can be significantly facilitated through the use of microneedle processing techniques even when the large-cap drugs are not loaded onto microneedles, not coated on microneedles, and / or constitute part of microneedles. Similarly, as will be understood by those skilled in the art reading this specification, this disclosure teaches that the delivery of large-cap drugs described herein can be significantly facilitated through the use of microneedle processing techniques (particularly through the use of MSCs) without leaving microneedles in the skin (e.g., by detaching them and / or otherwise retaining and / or disintegrating them in situ). For example, those skilled in the art will understand that the technology provided can avoid problems related to the long-term stability of major drugs required for commercially viable products and can achieve effective delivery without skin-disintegrating agents and / or microneedles remaining in the skin, which may result in inflammation, irritation, and / or allergic reactions. Indeed, in the examples and elsewhere, the disclosure expressly teaches that MSCs performed using microneedles that do not contain botulinum toxin facilitate the transdermal delivery of botulinum toxin from topical (e.g., creams, ointments) compositions, particularly compositions containing macro or nanoemulsions.

[0143] In some embodiments, the disclosure teaches that particularly advantageous results are achieved when the microneedle treatment technique is combined with a macroemulsion composition. In some embodiments, the microneedle treatment technique is combined with a lotion, cream, or liquid composition which may later become a macroemulsion composition or contain one. In some embodiments, the provided technique does not utilize skin-disintegrating techniques such as chemical penetration enhancers.

[0144] In some embodiments, the present invention utilizes macroemulsion compositions containing large-cap drugs that are particularly effective and / or useful in medical situations, for example, for therapeutic purposes. In some embodiments, certain macroemulsion compositions are particularly effective and / or useful for topical administration of drugs to subjects requiring it. In some embodiments, the macroemulsion composition may contain one or more large-cap drugs.

[0145] In some embodiments, the macroemulsion can be formulated into a composition suitable for topical administration to the skin. In some embodiments, the composition suitable for topical administration may be a lotion, cream, powder, ointment, liniment, gel, or drop.

[0146] In some embodiments, the macroemulsion formulation comprises water, medium-chain triglycerides, Span 65, polysorbate 80, methylparaben, and propylparaben.

[0147] In some embodiments, the provided composition comprises a mixture of the provided macroemulsion composition and one or more pharmaceutically acceptable excipients. In some embodiments, the cream and / or lotion formulation comprises a mixture of the provided macroemulsion composition and / or saline solution.

[0148] In some embodiments, the provided composition comprises a macroemulsion composition containing one or more large-capacity agents. In some embodiments, the provided composition is a cream and / or lotion formulation. In some embodiments, the provided cream and / or lotion formulation comprises a macroemulsion composition. In some embodiments, the composition comprises a provided macroemulsion composition but is not a cream and / or lotion formulation. In some embodiments, a preferred composition is formulated into a cream and / or lotion but does not contain a macroemulsion composition.

[0149] In some embodiments, the provided composition comprises a mixture of the provided macroemulsion composition and one or more pharmaceutically acceptable excipients for topical and / or transdermal administration (e.g., by lotion, cream, powder, ointment, liniment, gel, drop, etc.).

[0150] 2. Nanoemulsion In some embodiments, the disclosure provides strategies for using microneedle treatment to “modify” the skin to which a transdermal product has been, is being, or will be applied. Studies analyzing the transdermal delivery of small molecules (specifically, short hydrophilic peptides having molecular weights in the range of 400–1000 Da) have shown that “skin penetration of peptides depends on their molecular weight and decreases as molecular weight increases” (Zhang, S., et al., “Enhanced delivery of hydrophilic peptides in Despite previous reports suggesting that such strategies are likely only useful for drugs with low molecular weights, this disclosure provides the insight that such microneedle preparations can, surprisingly, offer significant benefits in facilitating the transdermal delivery of large drugs (e.g., those with molecular weights of approximately 100 kDa or greater).

[0151] This disclosure provides insight that the effective and rapid (i.e., administration within minutes) transdermal delivery of large molecules by such liquid nanoemulsion compositions can, surprisingly, be improved by combining nanoemulsion administration with microneedle skin conditioning (MSC).

[0152] This disclosure particularly demonstrates that microneedle processing techniques (e.g., microneedle preparation of the skin) can significantly enhance the transdermal delivery of large-capacity drugs (e.g., botulinum toxin and / or antibody drugs), especially when used in conjunction with nanoemulsion technology. The specific nanoemulsion compositions covered include water-in-oil and oil-in-water nanoemulsions characterized by droplet sizes ranging from about 1 nm to about 300 nm in diameter, oil-to-aqueous dispersion ratios ranging from about 0.01:1 to about 20:1, oil-to-surfactant ratios ranging from about 0.1 to about 40, and / or zeta potentials ranging from about -80 mV to about +80 mV.

[0153] In some embodiments, the provided nanoemulsion composition contains oil and surfactant in a ratio ranging from about 0.1:1 to about 2:1. In some embodiments, the provided nanoemulsion composition contains oil and surfactant in a ratio ranging from about 0.1:1 to about 1:1. In some embodiments, the provided nanoemulsion composition contains oil and surfactant in a ratio ranging from about 0.5:1 to about 1:1. In some embodiments, the provided nanoemulsion composition contains oil and surfactant in a ratio ranging from about 0.5:1 to about 1:1.5. In some embodiments, the provided nanoemulsion composition contains oil and surfactant in a ratio of about 0.1:1, about 0.15:1, about 0.2:1, about 0.25:1, about 0.3:1, about 0.35:1, about 0.4:1, about 0.45:1, about 0.5:1, about 0.5:1, about 0.55:1, about 0.6:1, about 0.65:1, about 0.7:1, about 0.75:1, about 0.8:1, about 0.85:1, about 0.9:1, about 0.95:1, or about 1:1. In some embodiments, the provided nanoemulsion composition contains oil and surfactant in a ratio of about 0.67:1.

[0154] In some embodiments, the aqueous dispersion medium (e.g., water, buffer solution, salt solution, etc.) and the surfactant are used in a ratio ranging from 0.01 to 20. In some embodiments, the aqueous dispersion medium (e.g., water, buffer solution, salt solution, etc.) and the surfactant are used in a ratio ranging from 0.1 to 20. In some embodiments, the aqueous dispersion medium (e.g., water, buffer solution, salt solution, etc.) and the surfactant are used in a ratio ranging from 0.5 to 10. In some embodiments, the aqueous dispersion medium (e.g., water, buffer solution, salt solution, etc.) and the surfactant are used in a ratio ranging from 0.5 to 1. In some embodiments, the ratio of the aqueous dispersion medium (e.g., water, buffer solution, salt solution, etc.) to the surfactant is approximately 0.01:1, approximately 0.02:1, approximately 0.03:1, approximately 0.04:1, approximately 0.05:1, approximately 0.06:1, approximately 0.07:1, approximately 0.08:1, approximately 0.0:1, approximately 0.1:1, approximately 0.2:1, approximately 0.3:1, approximately 0.4:1, approximately 0.5:1, approximately 1:1, approximately 2:1, approximately 3:1, approximately 4:1, approximately 5:1, approximately 6:1, approximately 7:1, approximately 8:1, approximately 9:1, or 10:1. In some embodiments, the ratio of surfactant to water is approximately 0.5:1, approximately 1:1, approximately 2:1, approximately 3:1, approximately 4:1, approximately 5:1, approximately 6:1, approximately 7:1, approximately 8:1, approximately 9:1, approximately 10:1, approximately 11:1, approximately 12:1, approximately 13:1, approximately 14:1, approximately 15:1, approximately 16:1, approximately 17:1, approximately 18:1, approximately 19:1, or approximately 20:1. In some embodiments, the aqueous dispersion medium (e.g., water, buffer, salt solution, etc.) and the surfactant are used in a ratio ranging from 0.5 to 2. In some embodiments, the ratio of aqueous dispersion medium (e.g., water, buffer, salt solution, etc.) to surfactant is approximately 0.5:1, approximately 1:1, or approximately 2:1. In some embodiments, the ratio of surfactant to aqueous dispersion medium (e.g., water, buffer, salt solution, etc.) is approximately 0.5:1, approximately 1:1, or approximately 2:1. In some embodiments, the ratio of the aqueous dispersion medium (e.g., water, buffer solution, salt solution, etc.) to the surfactant is approximately 1:1.In some embodiments, compositions utilizing such a ratio of aqueous dispersion medium (e.g., water, buffer solution, salt solution, etc.) to surfactant include water-in-oil emulsions.

[0155] In some embodiments, droplets in the nanoemulsion composition have diameters (e.g., average and / or median diameters) in the range of about 10 nm to about 300 nm, about 10 nm to about 200 nm, about 10 nm to about 150 nm, about 10 nm to about 130 nm, about 10 nm to about 120 nm, about 10 nm to about 115 nm, about 10 nm to about 110 nm, about 10 nm to about 100 nm, or about 10 nm to about 90 nm. In some embodiments, droplets in the nanoemulsion composition have diameters (e.g., average and / or median diameters) in the range of 1 nm to 300 nm, 1 nm to 200 nm, 1 nm to 150 nm, 1 nm to 120 nm, 1 nm to 100 nm, 1 nm to 75 nm, 1 nm to 50 nm, or 1 nm to 25 nm. In some embodiments, droplets in the nanoemulsion composition have diameters (e.g., average and / or median diameters) of 1 nm to 15 nm, 15 nm to 200 nm, 25 nm to 200 nm, 50 nm to 200 nm, or 75 nm to 200 nm.

[0156] In some embodiments, the total droplet distribution is contained within a specific range of droplet diameter sizes. In some embodiments, less than 50%, 25%, 10%, 5%, or 1% of the total droplet distribution is outside a specific range of droplet diameter sizes. In some embodiments, less than 1% of the total droplet distribution is outside a specific range of droplet diameter sizes. In some embodiments, the nanoemulsion composition substantially does not contain droplets having diameters greater than 300 nm, 250 nm, 200 nm, 150 nm, 120 nm, 100 nm, 75 nm, 50 nm, or 25 nm. In some embodiments, less than 50%, 25%, 10%, 5%, or 1% of the total droplet distribution has diameters greater than 300 nm, 250 nm, 200 nm, 150 nm, 120 nm, 100 nm, 75 nm, 50 nm, or 25 nm.

[0157] In some embodiments, droplets in the nanoemulsion composition have an average droplet size of about 300 nm, about 250 nm, about 200 nm, about 150 nm, about 130 nm, about 120 nm, about 115 nm, about 110 nm, about 100 nm, about 90 nm, or less than about 50 nm. In some embodiments, the average droplet size is in the range of about 10 nm to about 300 nm, about 50 nm to about 250 nm, about 60 nm to about 200 nm, about 65 nm to about 150 nm, or about 70 nm to about 130 nm. In some embodiments, the average droplet size is about 80 nm to about 110 nm. In some embodiments, the average droplet size is about 90 nm to about 100 nm.

[0158] In some embodiments, the nanoemulsion droplets have a zeta potential in the range of -80mV to +80mV. In some embodiments, the nanoemulsion droplets have a zeta potential in the range of -50mV to +50mV. In some embodiments, the nanoemulsion droplets have a zeta potential in the range of -25mV to +25mV. In some embodiments, the nanoemulsion droplets have a zeta potential in the range of -10mV to +10mV. In some embodiments, the nanoemulsion droplets have a zeta potential of about -80mV, about -70mV, about -60mV, about 50mV, about -40mV, about -30mV, about -25mV, about -20mV, about -15mV, about -10mV, or about -5mV. In some embodiments, the nanoemulsion droplets have a zeta potential of approximately +50mV, approximately +40mV, approximately +30mV, approximately +25mV, approximately +20mV, approximately +15mV, approximately +10mV, or approximately +5mV. In some embodiments, the nanoemulsion droplets have a zeta potential of approximately 0mV.

[0159] This disclosure provides a remarkably effective technique for the transdermal delivery of large-cap drugs (e.g., botulinum toxin, antibodies, etc.). In particular, this disclosure teaches that the transdermal delivery of such drugs can be significantly facilitated through the use of microneedle processing techniques without any other disintegrating measures. Thus, the techniques provided can achieve effective delivery without the inflammation, irritation, and / or allergic reactions that often occur with the use of skin-disintegrating agents. As will be understood by those skilled in the art reading this specification, this disclosure teaches that the transdermal delivery of large-cap drugs can be significantly facilitated through the use of microneedle processing techniques even when the large-cap drugs are not loaded onto microneedles, not coated on microneedles, and / or constitute part of microneedles. Similarly, as will be understood by those skilled in the art reading this specification, this disclosure teaches that the delivery of large-cap drugs described herein can be significantly facilitated through the use of microneedle processing techniques (and in particular through the use of MSCs) without leaving microneedles in the skin (e.g., by detaching them and / or otherwise retaining and / or disintegrating them in situ). For example, those skilled in the art will understand that the technology provided can avoid problems related to the long-term stability of major drugs required for commercially viable products and can achieve effective delivery without skin-disintegrating agents and / or microneedles remaining in the skin, which may result in inflammation, irritation, and / or allergic reactions. Indeed, in the examples and elsewhere, the disclosure expressly teaches that MSCs performed using microneedles that do not contain botulinum toxin facilitate the transdermal delivery of botulinum toxin from topical (e.g., creams, ointments) compositions, particularly compositions containing macro or nanoemulsions.

[0160] In some embodiments, the disclosure teaches that particularly advantageous results are achieved when the microneedle treatment technique is combined with a nanoemulsion composition. In some embodiments, the microneedle treatment technique is combined with a lotion, cream, or liquid composition which may later become a nanoemulsion composition or contain one. In some embodiments, the provided technique does not utilize skin-disintegrating techniques such as chemical penetration enhancers.

[0161] The findings presented herein are particularly surprising given reports that transdermal delivery of solid nanoparticles of a size comparable to that of droplets in the nanoemulsion compositions used herein (e.g., 10⁵ ± 2.92 nm) does not effectively deliver (or facilitate) even small molecule drugs transdermally through the skin. For example, Gomaa et al. described a study in which a solution of rhodamine dye (molecular weight 479 Da) encapsulated in PLGA nanoparticles was applied to skin pre-prepared by microneedle treatment to evaluate skin penetration. See Gomaa, Y., et al, “Effect of microneedle treatment on the skin permeation of a nanoencapsulated dye.” J Pharm Pharmacol. 2012 November;64(11):1592-1602. The data showed that very small amounts of dye began to penetrate the skin after 6 hours of continuous application; no significant increase in penetration was observed until the skin was treated continuously for 24 hours. The researchers explained that there is a growing consensus that nanoparticles (NPs) cannot normally penetrate the stratum corneum but can be well deposited within hair follicles. Therefore, before this disclosure, those skilled in the art would anticipate that the use of microneedle treatment techniques with nano-sized vehicles would not be able to effectively deliver even small molecule drugs (e.g., rhodamine dye) transdermally; and that delivery of large-molecule drugs would certainly be impossible. However, this disclosure demonstrates that microneedle treatment can significantly facilitate the transdermal delivery of large-molecule drugs, particularly when used in conjunction with nanoemulsion systems.

[0162] In particular, this disclosure demonstrates that microneedling techniques can facilitate transdermal delivery (e.g., large drugs, especially from nanoemulsion compositions) when no other disintegrants are used (i.e., no chemical penetration enhancers are used, and no other techniques are used to disintegrate or puncture skin structures). Previous studies of transdermal delivery of drugs about the size of botulinum toxin (i.e., about 150 kDa) using microneedling have reported that delivery is unsuccessful unless additional treatment is applied to disintegrate the skin. For example, U.S. Patent Publication 2010 / 0196445 reports that botulinum toxin is not effectively delivered from pre-coated microneedles unless skin digestive enzymes are also applied so that skin structures are disintegrated at the microneedling site.

[0163] This disclosure demonstrates, in particular, that microneedle processing techniques can achieve transdermal delivery (e.g., of large-capacity drugs from microemulsion and nanoemulsion compositions) when microneedle coating or loading is not utilized and / or when the microneedles are not designed to remain in the skin. In particular, as already described, this disclosure understands that such microneedle coating or loading may not be commercially viable, at least due to the instability of the botulinum coating or loading material. For example, Johnson, E., et al. According to al., "Botulinum toxin is highly susceptible to denaturation due to surface denaturation, heat, and alkaline conditions. Lyophilization or freeze-drying of botulinum toxin is the most economically rational and practical method of distributing the product in a stable form that is readily available to clinicians." U.S. Patent No. 5,512,547. Also, as will be understood by those skilled in the art reading this specification, the techniques described herein have certain advantages, including the fact that it is not necessary to leave microneedles in or with the tissue. For example, those skilled in the art will understand that leaving microneedles in the skin may carry the risk of skin irritation, inflammation, allergic reactions, and / or cosmetically undesirable scarring. In contrast to the present invention, techniques such as those described in U.S. Patent No. 2017 / 0209553 utilize a microneedle array designed in which botulinum toxin is loaded into the needle and the microneedles detach and penetrate the skin (as per U.S. Patents No. 2017 / 0209553 and 2016 / 0263362).

[0164] This disclosure teaches that, despite the expectation that MSCs are only useful in facilitating the transdermal delivery of small compounds, the already highly effective transdermal delivery of large drugs by nanoemulsion compositions can be dramatically enhanced by combining them with microneedle technology. Furthermore, this disclosure is particularly surprising considering that microneedle preparations combined with encapsulation of small molecule drugs in solid nanoparticles, as described by Gomaa, provided only a small amount of penetration as early as 6 hours after administration, and no material penetration was observed until 24 hours after administration.

[0165] In some embodiments, a nanoemulsion formulation containing a large drug is administered in conjunction with microneedle preparation using a solid microneedle. In some embodiments, site MSC is performed before applying the nanoemulsion formulation containing the large drug to that site (e.g., before a specific application and / or before each application). In some embodiments, site MSC is performed after applying the nanoemulsion formulation containing the large drug to that site. In some embodiments, site MSC and application of the nanoemulsion formulation containing the large drug to that site occur substantially simultaneously. In some embodiments, the macroemulsion formulation containing the large drug is not injected via one or more microneedles. In some embodiments, the microneedle is part of an array of microneedles. In some embodiments, the microneedle may have a length of about 1 μm to about 4,000 μm. In some embodiments, the microneedle may have a length of about 1 μm to about 2,000 μm. In some embodiments, the microneedle may have a length of about 50 μm to about 400 μm. In some embodiments, the microneedles may have a length of approximately 800 μm to approximately 1500 μm.

[0166] In some embodiments, the present invention utilizes nanoemulsion compositions containing large-cap drugs that are particularly effective and / or useful in medical situations, for example, for therapeutic purposes. In some embodiments, certain nanoemulsion compositions are particularly effective and / or useful for topical administration of drugs to targets requiring it. In some embodiments, the nanoemulsion composition may contain one or more large-cap drugs. Exemplary nanoemulsion compositions and methods for their production are described, for example, in WO2012 / 103035, the disclosure of which is incorporated in whole by reference.

[0167] In some embodiments, the nanoemulsion can be formulated into a composition suitable for topical administration. In some embodiments, the composition suitable for topical administration may be a lotion, cream, powder, ointment, liniment, gel, or drop.

[0168] In some embodiments, the nanoemulsion formulation comprises water, medium-chain triglycerides, polysorbate 80, methylparaben, and propylparaben. Exemplary premixes, not intended to be limiting, are provided in Table 2. [Table 2] *The buffer solution contained 0.199% gelatin, 0.398% dibasic sodium phosphate, and 99.4% purified water (w / w), and the pH was adjusted to 6.0 ± 0.2 with hydrochloric acid. **Those skilled in the art may, in consideration of this specification, make reasonable adjustments to this and other components depending on the volume, weight, and / or dosage of the large-capacity drug used.**

[0169] These compositions are particularly useful in that they can be used to deliver a drug to a subject in need via topical and / or transdermal administration (e.g., by lotion, cream, powder, ointment, liniment, gel, drop, etc.). In some embodiments, the provided cream and / or lotion formulations can be administered to a subject in need via topical and / or transdermal administration (e.g., by lotion, cream, powder, ointment, liniment, gel, drop, etc.). In some embodiments, the provided nanoemulsion composition can be formulated into a cream and / or lotion formulation. In some embodiments, the provided cream and / or lotion formulation containing the nanoemulsion composition may be useful and / or effective for topical administration to a subject. In some embodiments, the provided nanoemulsion composition can be mixed with one or more cream components in a cream formulation (e.g., the provided cream formulation) and / or saline solution for preparing a pharmaceutical composition.

[0170] The present invention encompasses the recognition that emulsion compositions (e.g., macroemulsion compositions and nanoemulsion compositions) can be formulated into cream and / or lotion formulations for administration to a subject. The present invention also encompasses the recognition that the provided cream and / or lotion formulations may be particularly useful for formulating emulsions, such as those described herein, for administration to a subject. Exemplary nanoemulsion formulations are provided in Table 3, not intended to be limiting. [Table 3] *Those skilled in the art may, in consideration of this specification, make reasonable adjustments to this and other components depending on the volume, weight, and / or dosage of the large-capacity drug used.

[0171] In some embodiments, the provided composition comprises a mixture of the provided nanoemulsion composition and one or more pharmaceutically acceptable excipients. In some embodiments, the cream and / or lotion formulation comprises a mixture of the provided nanoemulsion composition and / or saline solution.

[0172] In some embodiments, the provided composition comprises the provided nanoemulsion composition. In some embodiments, the provided composition is a cream and / or lotion formulation. In some embodiments, the provided cream and / or lotion formulation comprises the nanoemulsion composition. In some embodiments, the composition comprises the provided nanoemulsion composition but is not a cream and / or lotion formulation. In some embodiments, a preferred composition is formulated into a cream and / or lotion but does not contain the nanoemulsion composition.

[0173] In some embodiments, the provided composition comprises a mixture of the provided nanoemulsion composition and one or more pharmaceutically acceptable excipients for topical and / or transdermal administration (e.g., by lotion, cream, powder, ointment, liniment, gel, drop, etc.).

[0174] In some embodiments, for nanoemulsion compositions comprising known therapeutic agents and / or independently active biological agents, such nanoemulsion compositions are prepared, configured, and administered in combination with MSCs such that a sufficient amount of the therapeutic agent is delivered to a desired target site (e.g., the epidermis and / or dermal structure) to treat a disease or disorder. In some embodiments, the provided nanoemulsion compositions are prepared and configured (e.g., by the selection and / or combination of agents, the structure of the composition, etc.) to achieve a desired therapeutic effect upon administration to the skin. In some embodiments, the provided nanoemulsion compositions are prepared and configured so that they do not induce undesirable clinical effects inside and / or outside the desired site of action (e.g., the surface of the skin, the dermis, etc.). In some embodiments, the provided nanoemulsion compositions are prepared, configured, and administered in combination with MSCs such that they have a systemic effect.

[0175] In some embodiments, the provided composition may be formulated and delivered in combination with MSCs so as to achieve systemic delivery; in some embodiments, the provided composition may be formulated and / or delivered so as to achieve topical but non-systemic delivery.

[0176] This disclosure specifically demonstrates the effective and efficient delivery of therapeutic agents (and, in particular, large-scale biologics such as botulinum toxin or antibody drugs) to the dermis using compositions provided in combination with MSCs. For example, in some embodiments, the present invention provides methods comprising the administration of compositions described herein without clinically significant side effects. For example, when local delivery is intended, clinically significant side effects include, but are not limited to, undesirable systemic side effects, damage to nerve tissue beneath the dermis (e.g., nerve paralysis), undesirable effects on muscles (e.g., muscle paralysis), and / or undesirable blood levels of the therapeutic agent. Exemplary formulations of botulinum nanoemulsion premixes are provided in Table 4, which are not intended to limit the scope of such formulations. [Table 4] *The buffer solution contained 0.199% gelatin, 0.398% dibasic sodium phosphate, and 99.4% purified water (w / w), and the pH was adjusted to 6.0 ± 0.2 with hydrochloric acid.

[0177] Diseases, disorders, and pathological conditions The present invention provides techniques for treating and / or preventing any variety of systemic or cutaneous diseases, disorders, and / or conditions. In some embodiments, the present invention provides techniques for treating and / or preventing diseases, disorders, or conditions related to the activity of sweat glands and / or sebaceous glands. In some embodiments, the present invention provides techniques for treating and / or preventing diseases, disorders, or conditions related to infections. In some embodiments, the present invention provides techniques for treating and / or preventing diseases, disorders, or conditions related to inflammation. In some embodiments, the present invention provides techniques for treating and / or preventing diseases, disorders, or conditions related to inflammation. In some embodiments, the present invention provides techniques for treating and / or preventing diseases, disorders, or conditions related to cancer. In some embodiments, the present invention provides techniques for treating and / or preventing systemic diseases, disorders, or conditions. In some embodiments, the present invention provides techniques for treating and / or preventing autoimmune diseases, disorders, or conditions. In some embodiments, the present invention provides techniques for treating and / or preventing diseases, disorders, or conditions related to the epidermal and / or dermal levels of the skin.

[0178] In some embodiments, the present invention addresses acne, excessive sweating, body odor, hyperhidrosis, bromhidrosis, chromatohidrosis, rosacea, alopecia, psoriasis, actinic keratosis, eczematous dermatitis (e.g., atopic dermatitis), sebum overproduction disorders (e.g., seborrhea, seborrheic dermatitis), burns, Raynaud's phenomenon, lupus erythematosus, hyperpigmentation disorders (e.g., melanosis), hypopigmentation disorders (e.g., vitiligo), skin cancer (e.g., squamous cell carcinoma, basal cell carcinoma), skin infections (e.g., bacterial infections, viral infections, fungal infections), and facial wrinkles (e.g., forehead, frown lines, wrinkles and / or periorbital wrinkles). The present invention provides techniques for treating and / or preventing one or more of the following: wrinkles (including those in the surrounding area), headaches, unsightly facial expressions (e.g., due to overactivity of the underlying facial muscle tissue), neck muscles, multifunctional facial muscles, hypercontracted facial muscles, platysma girdle, neuromuscular disorders and conditions including muscle spasms and / or contractures (including various forms of facial paralysis, cerebral palsy, blepharospasm, facial contractures), dystonia, benign prostatic hyperplasia, headaches, strabismus, hemifacial spasm, tremors, spasticity such as that resulting from multiple sclerosis, posterior orbital muscles, various ophthalmic and urological conditions (e.g., penile and / or bladder disorders), and / or combinations thereof.

[0179] In some embodiments, the present invention provides techniques for treating and / or preventing rheumatoid arthritis. In some embodiments, the present invention provides techniques for treating and / or preventing psoriatic arthritis. In some embodiments, the present invention provides techniques for treating and / or preventing osteoarthritis.

[0180] In some embodiments, the present invention provides techniques for treating and / or preventing lupus erythematosus. In some embodiments, lupus erythematosus is systemic, discoid, drug-induced, or neonatal.

[0181] In some embodiments, the present invention provides techniques for treating and / or preventing Crohn's disease. In some embodiments, the present invention provides techniques for treating and / or preventing inflammatory bowel disease. In some embodiments, the present invention provides techniques for treating and / or preventing ulcerative colitis.

[0182] In some embodiments, the present invention provides techniques for treating and / or preventing lung disorders. In some embodiments, the lung disorder may be asthma or chronic obstructive pulmonary disease.

[0183] In some embodiments, the present invention provides techniques for treating and / or preventing amyloidosis. In some embodiments, amyloidosis is systemic or cutaneous.

[0184] In some embodiments, the present invention provides techniques for treating and / or preventing cancer. In some embodiments, the cancer is of the skin, blood, breast, colon, or lung.

[0185] In some embodiments, the present invention provides techniques for treating and / or preventing dyslipidemia. In some embodiments, dyslipidemia is hypercholesterolemia.

[0186] In some embodiments, the present invention provides techniques for treating and / or preventing infectious diseases. In some embodiments, the infectious disease is C. difficile or Staphylococcus, or is caused by them.

[0187] In some embodiments, the present invention provides techniques for treating and / or preventing pain. In some embodiments, the pain is related to arthritis. In some embodiments, the arthritis is rheumatoid arthritis, psoriatic arthritis, or osteoarthritis.

[0188] In some embodiments, the present invention provides techniques for treating and / or preventing neurological conditions. In some embodiments, the neurological conditions are Alzheimer's disease, Parkinson's disease, or stroke.

[0189] In some embodiments, the present invention follows a drug regimen sufficient to achieve at least about 20% reduction in the degree and / or incidence of the relevant skin condition; in some embodiments, according to a drug regimen sufficient to achieve at least about 25% reduction; in some embodiments, according to a drug regimen sufficient to achieve at least about 30% reduction; in some embodiments, according to a drug regimen sufficient to achieve at least about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, The administration includes at least one of the provided compositions administered in combination with MSCs, according to a dosage regimen sufficient to achieve a reduction of approximately 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, or more.

[0190] In some embodiments, the present invention follows a dosage regimen sufficient to achieve a reduction of at least about 20% in the degree and / or incidence of the relevant skin condition in a specific proportion of the population of patients administered the composition; in some embodiments, a dosage regimen sufficient to achieve a reduction of at least about 25% in a specific proportion of the population of patients administered the composition; in some embodiments, a dosage regimen sufficient to achieve a reduction of at least about 30% in a specific proportion of the population of patients administered the composition; in some embodiments, at least about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, and about 4% in a specific proportion of the population of patients administered the composition. 0%, approximately 41%, approximately 42%, approximately 43%, approximately 44%, approximately 45%, approximately 46%, approximately 47%, approximately 48%, approximately 49%, approximately 50%, approximately 51%, approximately 52%, approximately 53%, approximately 54%, approximately 55%, approximately 56%, approximately 57%, approximately 58%, approximately 59%, approximately 60%, approximately 61%, approximately 62%, approximately 63%, approximately 64%, approximately 65%, approximately 66%, approximately 67%, approximately 68%, approximately 69%, approximately 70%, approximately 71%, approximately 72% The administration of at least one of the provided compositions administered in combination with MSCs, according to a dosage regimen sufficient to achieve a reduction of approximately 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, or more. In some embodiments, a particular percentage of the patient population administered with the composition is at least approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. To give a few illustrative examples, in some embodiments, the present invention includes the administration of at least one of the provided compositions according to a dosage regimen sufficient to achieve a reduction of at least about 20% in the degree and / or incidence of the associated skin condition in at least about 50% of the patient population to which the composition is administered.In some embodiments, the present invention includes the administration of at least one of the provided compositions in accordance with a dosage regimen sufficient to achieve a reduction of at least about 30% in the degree and / or morbidity of the associated skin condition in at least about 50% of a population of patients to whom the composition is administered.

[0191] The present invention provides techniques for treating and / or preventing skin conditions, comprising administering a provided composition in combination with MSCs to subjects suffering from, susceptible to, and / or exhibiting symptoms of skin conditions. In some embodiments, the provided compositions for the treatment of skin conditions described herein are formulated for any route of administration described herein. In some embodiments, the provided compositions are formulated for topical administration. In some embodiments, the provided compositions are formulated to be appropriate for the condition being treated, such as creams, liniments, lotions, gels, shampoos, conditioners, sunscreens, deodorants, and / or antiperspirants (e.g., roll-ons, solid sticks, gels, creams, aerosols, etc.).

[0192] In some embodiments, such a provided composition is administered topically in combination with MSCs to the affected area (e.g., the armpit, hand, foot, scalp, hair follicles, face, neck, back, arm, chest, leg, groin, or groin, as appropriate for the specific condition being treated). In some embodiments, topical administration is achieved by topical administration in combination with MSCs.

[0193] Compositions and Formulations As described herein, the present invention provides and / or utilizes compositions comprising one or more large-capacity agents for administration in combination with MSCs. In some embodiments, the compositions provided may be formulated for topical and / or transdermal delivery (e.g., as lotions, creams, liniments, ointments, powders, gels, drops, etc.). In some embodiments, the compositions provided may be or comprise nanoemulsions. In some embodiments, the compositions provided may be or comprise macroemulsions.

[0194] Formulations of the provided composition may be prepared, for example, by any suitable method known or to be developed in the field of pharmacology. Generally, such preparation methods include associating the provided composition with one or more excipients, and then, if necessary and / or desirable, forming and / or packaging it into an administration-appropriate form, for example, as or in single or multiple dose units.

[0195] In some embodiments, the composition may be prepared, packaged, and / or sold in large quantities as a single unit dose and / or as multiple single doses. As used herein, “unit dose” refers to an individual amount of a pharmaceutical composition comprising a predetermined amount of the composition to be provided. The amount of the composition to be provided is generally equal to the dosage of the composition to be administered to a subject and / or a favorable proportion of such dosage, for example, half or one-third of such dosage.

[0196] In some embodiments, suitable excipients for use in a composition (e.g., a pharmaceutically and / or cosmetically acceptable composition) may include, for example, one or more excipients, such as solvents, dispersion media, granulation media, diluents or other liquid vehicles, dispersion or suspension aids, surfactants and / or emulsifiers, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, splitting agents, binders, preservatives, buffers, etc. (as suitable for the desired specific dosage form). In some embodiments, excipients such as cocoa butter and / or suppository waxes, colorants, coatings, sweeteners, flavorings, and / or fragrances may be used. Remington's The Science and Practice of Pharmacy, 21 st Edition, ARGennaro (Lippincott, Williams & Wilkins, Baltimore, MD, 2005; incorporated herein by reference) discloses various excipients used in formulating pharmaceutical compositions and known techniques for their preparation.

[0197] In some embodiments, the appropriate excipient (e.g., a pharmaceutically and / or cosmetically acceptable excipient) is at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% pure. In some embodiments, the excipient is approved by the U.S. Food and Drug Administration. In some embodiments, the excipient is pharmaceutical grade. In some embodiments, the excipient meets the standards of the United States Pharmacopeia (USP), European Pharmacopoeia (EP), British Pharmacopoeia, and / or other international pharmacopoeias.

[0198] In some embodiments, the compositions provided are formulated as creams, liniments, ointments, oils, foams, sprays, lotions, liquids, powders, thickened lotions, or gels (for example, formulated for transdermal delivery as described herein). Certain exemplary such formulations may be prepared as cosmetic formulations such as skin softeners, nourishing lotion-type emulsions, cleansing lotions, cleansing creams, skin milks, emollient lotions, massage creams, emollient creams, cosmetic bases, face packs or face gels, cleansing formulations such as shampoos, conditioners, body washes, hair tonics, or soaps, or skin compositions such as lotions, ointments, gels, creams, liniments, patches, deodorants, or sprays. In some embodiments, compositions for topical administration are not formulated for administration to mucous membranes (for example, unsuitable for application to mucous membranes and / or not formulated to deliver a suitable amount of a large drug to or over mucous membranes).

[0199] treatment area The technology of the present invention is suitable for both human and veterinary use. Subjects suffering from any disorder that would benefit from topical application of an active agent can be treated with the disclosed technology for transdermal drug delivery.

[0200] A preferred site for MSC is a preferred administration site. In some embodiments, the administration site is the skin covering the muscle or muscle group of the target. In some embodiments, the site is hairless. In some embodiments, the site is on the torso. In some embodiments, the site is on the back. In some embodiments, the site is on the chest. In some embodiments, the site is on the buttocks. In some embodiments, the site is in the groin. In some embodiments, the site is in the inguinal region. In some embodiments, the site is on the head. In some embodiments, the site is on the scalp. In some embodiments, the site is on the face. In some embodiments, the site is on the neck. In some embodiments, the site is on the chest. In some embodiments, the site is under the armpit. In some embodiments, the site is in the axilla. In some embodiments, the site is on the hand. In some embodiments, the site is on the foot. In some embodiments, the site is on the arm. In some embodiments, the site is on the leg. In some embodiments, the site is not mucous membrane.

[0201] In some embodiments, the site is affected by a skin condition. In some embodiments, the site is the skin covering a muscle or muscle group affected by a neuromuscular condition. In some embodiments, the length of the microneedle used in MSC is adjusted based on the thickness of the skin at the treatment site.

[0202] In some embodiments, the MSC includes one impression of an MN or an MN array. In some embodiments, the MSC includes two impressions of an MN or an MN array. In some embodiments, the MSC includes three impressions of an MN or an MN array. In some embodiments, the MSC includes four impressions of an MN or an MN array. In some embodiments, the MSC includes five impressions of an MN or an MN array. In some embodiments, the MSC includes six impressions of an MN or an MN array. In some embodiments, the MSC includes seven impressions of an MN or an MN array. In some embodiments, the MSC includes eight impressions of an MN or an MN array. In some embodiments, the MSC includes nine impressions of an MN or an MN array. In some embodiments, the MSC includes ten impressions of an MN or an MN array. In some embodiments, the MSC includes eleven impressions of an MN or an MN array. In some embodiments, the MSC includes twelve impressions of an MN or an MN array. In some embodiments, the MSC includes thirteen impressions of an MN or an MN array. In some embodiments, the MSC includes fourteen impressions of an MN or an MN array. In some embodiments, the MSC includes 15 impressions of the MN or MN array. In some embodiments, the MSC includes 16 impressions of the MN or MN array. In some embodiments, the MSC includes 17 impressions of the MN or MN array. In some embodiments, the MSC includes 18 impressions of the MN or MN array. In some embodiments, the MSC includes 19 impressions of the MN or MN array. In some embodiments, the MSC includes 20 impressions of the MN or MN array. In some embodiments, the MSC includes rotating the MN or MN array on the skin one or more times. In some embodiments, the MN array is rotated between impressions. In some embodiments, the MN array is not rotated between impressions. In some embodiments, impressions are made on the same site. In some embodiments, impressions are made on overlapping sites. In some embodiments, impressions are made on different sites. In some embodiments, impressions are made by stamping the MN array.In some embodiments, the impression is made by rotating a microneedle roller over the site one or more times. In accordance with established MN practices, in some embodiments, the skin impression of the MN array lasts for less than one second, or alternatively, in some embodiments, they may last for more than one second, for example, 30 seconds or more, 60 seconds or more, 2 minutes or more, 5 minutes or more, 10 minutes or more, 30 minutes or more, etc.

[0203] Administration The present invention provides a technology for delivering emulsion compositions (e.g., botulinum emulsion compositions or antibody drug emulsion compositions) for various applications, including, for example, cosmetic agents, nutritional supplements, and medical applications. Such emulsion compositions may contain one or more biological agents. In some embodiments, the emulsion composition contains botulinum toxin. In some embodiments, the emulsion composition contains an antibody drug. In some embodiments, the emulsion composition is a nanoemulsion composition and / or a macroemulsion composition.

[0204] The present invention provides a technique for treating a pathological condition or disorder using any of the compositions provided herein in combination with MSC (e.g., the provided emulsion composition; a cream and / or lotion formulation; a combination of the provided emulsion composition and a cream and / or lotion formulation; etc.).

[0205] In some embodiments, such a method includes administering a provided composition in combination with MSCs to patients suffering from and / or susceptible to a disease, condition, or disorder. In some embodiments, such a method includes administering a provided nanoemulsion composition in combination with MSCs to patients suffering from and / or susceptible to a disease, condition, or disorder relating to the dermis layer of the skin. In some embodiments, such a method includes administering an emulsion composition in combination with MSCs containing at least one known therapeutic agent and / or independently active biological agent to patients suffering from and / or susceptible to a disease, condition, or disorder. In some embodiments, such a method includes administering an emulsion composition formulated in a provided cream and / or lotion formulation and / or at least one known therapeutic agent and / or independently active biological agent in combination with MSCs to patients suffering from and / or susceptible to a disease, condition, or disorder. In some embodiments, such methods include the administration of a composition via topical and / or transdermal administration (e.g., by lotion, cream, powder, ointment, liniment, gel, drop, etc.) in combination with MSCs. Some embodiments further include the administration of an penetration enhancer. Some embodiments further include the administration of a non-irritating penetration enhancer.

[0206] In some embodiments, the present invention provides techniques for treating any pathological condition or disorder. In some embodiments, the present invention demonstrates that a predetermined composition described herein, combined with MSCs, can achieve controlled delivery of an active agent efficiently and specifically to a biologically relevant target site (e.g., a specific tissue, a location within the skin, a cell, etc.). In some embodiments, the present invention demonstrates the achievement of controlled delivery and / or therapeutic effect at a predetermined biologically relevant target site without significant side effects associated with delivery to other areas.

[0207] In some embodiments, the present invention provides techniques for treating pathological conditions or disorders related to epidermal and / or dermal structures (e.g., sweat glands, sebaceous glands, hair follicles, etc.). In some embodiments, the present invention demonstrates that the compositions provided herein (e.g., provided nanoemulsion compositions; cream and / or lotion formulations; combinations of provided nanoemulsion compositions and cream and / or lotion formulations; etc.) combined with MSCs can efficiently and specifically deliver active agents to the dermis, and that the compositions provided herein can exert a therapeutic effect on the target skin upon administration. In some embodiments, the present invention demonstrates dermal delivery and / or achievement of therapeutic effect without significant side effects associated with delivery to other areas (e.g., subcutaneous or extracutaneous structures and / or tissues other than the dermis). In some embodiments, the compositions provided herein (e.g., the emulsion compositions provided; cream and / or lotion formulations; combinations of the emulsion compositions provided and cream and / or lotion formulations; etc.) combined with MSCs can deliver active agents such as therapeutic agents (e.g., botulinum toxin, antibody drugs, etc.) transdermally.

[0208] The present invention provides a technique for treating a disease or disorder by administering to a patient a composition provided herein (e.g., a provided emulsion composition; a cream and / or lotion formulation; a combination of a provided emulsion composition and a cream and / or lotion formulation; etc.) in combination with MSCs. In some embodiments, the present invention provides a technique for treating a disease or disorder by topically administering to a patient a composition containing an emulsion composition provided in combination with MSCs as described herein.

[0209] In some embodiments, the large drug penetrates the skin within approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 minutes after administration. In some embodiments, the large drug penetrates the skin within approximately 5 to 60 minutes after administration. In some embodiments, the large drug penetrates the skin within approximately 5 to 12 minutes after administration. In some embodiments, the large drug penetrates the skin within approximately 5 to 15 minutes after administration. In some embodiments, the large drug penetrates the skin within approximately 15 to 30 minutes after administration. In some embodiments, the large drug penetrates the skin within approximately 1 hour after administration. In some embodiments, the large drug penetrates the skin within approximately 2 hours after administration. In some embodiments, the large drug penetrates the skin within approximately 3 hours after administration. In some embodiments, the large drug penetrates the skin within approximately 4 hours after administration. In some embodiments, the large drug penetrates the skin within approximately 5 hours after administration. In some embodiments, the large drug penetrates the skin within approximately 6 hours after administration.

[0210] In some embodiments, the large-capacity drug penetrates the skin layer within approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 minutes after administration. In some embodiments, the large-capacity drug penetrates the skin layer within approximately 5 to 60 minutes after administration. In some embodiments, the large-capacity drug penetrates the skin layer within approximately 5 to 12 minutes after administration. In some embodiments, the large-capacity drug penetrates the skin layer within approximately 5 to 15 minutes after administration. In some embodiments, the large-capacity drug penetrates the skin layer within approximately 15 to 30 minutes after administration. In some embodiments, the large-capacity drug penetrates the skin layer within approximately 1 hour after administration. In some embodiments, the large-capacity drug penetrates the skin layer within approximately 2 hours after administration. In some embodiments, the large-capacity drug penetrates the skin layer within approximately 3 hours after administration. In some embodiments, the large-capacity drug penetrates the skin layer within approximately 4 hours after administration. In some embodiments, the large-capacity drug penetrates the skin layer within approximately 5 hours after administration. In some embodiments, the large drug penetrates the skin layer within approximately 6 hours of administration.

[0211] In some embodiments, the large-capacity drug penetrates the uppermost layer of the skin within approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 minutes after administration. In some embodiments, the large-capacity drug penetrates the uppermost layer of the skin within approximately 5 to 60 minutes after administration. In some embodiments, the large-capacity drug penetrates the uppermost layer of the skin within approximately 5 to 12 minutes after administration. In some embodiments, the large-capacity drug penetrates the uppermost layer of the skin within approximately 5 to 15 minutes after administration. In some embodiments, the large-capacity drug penetrates the uppermost layer of the skin within approximately 15 to 30 minutes after administration. In some embodiments, the large-capacity drug penetrates the uppermost layer of the skin within approximately 1 hour after administration. In some embodiments, the large-capacity drug penetrates the uppermost layer of the skin within approximately 2 hours after administration. In some embodiments, the large-capacity drug penetrates the uppermost layer of the skin within approximately 3 hours after administration. In some embodiments, the large-capacity drug penetrates the uppermost layer of the skin within approximately 4 hours after administration. In some embodiments, the large-capacity drug penetrates the uppermost layer of the skin within approximately 5 hours of administration. In some embodiments, the large-capacity drug penetrates the uppermost layer of the skin within approximately 6 hours of administration.

[0212] In some embodiments, the large-capacity drug penetrates the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, within approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 minutes after administration. In some embodiments, the large-capacity drug penetrates the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, within approximately 5 to 60 minutes after administration. In some embodiments, the large-capacity drug penetrates the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, within approximately 5 to 12 minutes after administration. In some embodiments, the large-capacity drug penetrates the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, within approximately 5 to 15 minutes after administration. In some embodiments, the large-capacity drug penetrates the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, within approximately 15 to 30 minutes after administration. In some embodiments, the large-capacity drug penetrates the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, within approximately one hour of administration. In some embodiments, the large-capacity drug penetrates the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, within approximately two hours of administration. In some embodiments, the large-capacity drug penetrates the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, within approximately three hours of administration. In some embodiments, the large-capacity drug penetrates the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, within approximately four hours of administration. In some embodiments, the large-capacity drug penetrates the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, within approximately five hours of administration. In some embodiments, the large-capacity drug penetrates the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, within approximately six hours of administration.

[0213] kit In some embodiments, the present invention provides a pharmaceutical pack or kit comprising one or more emulsion compositions and one or more microneedle devices according to the present invention. In some embodiments, the pharmaceutical pack or kit comprises a preparation or pharmaceutical composition containing the composition provided in one or more containers optionally filled with one or more additional components of the pharmaceutical composition. In some embodiments, the pharmaceutical pack or kit comprises additional approved therapeutic agents for use in combination therapy (e.g., benzoyl peroxide for the treatment of acne; aluminum compounds for the treatment of hyperhidrosis; etc.). In some embodiments, a notice in the form prescribed by a government agency regulating the manufacture, use or sale of a pharmaceutical product, reflecting approval by an agency for manufacture, use or sale for human administration, may optionally be associated with such container(s).

[0214] In some embodiments, a kit containing a therapeutic agent is provided. In one but non-limiting example, the composition provided may be provided as a topical formulation and administered as a therapy in combination with the use of a microneedle processing device. Thus, a pharmaceutical dosage or instructions for self-administration may be provided in a kit for administration to an individual suffering from or at risk of suffering from a condition or disorder, such as one relating to the dermal level of the skin.

[0215] In some embodiments, the kit may include (i) a composition provided; (ii) at least one pharmaceutically acceptable excipient; and (iii) at least one device for microneedle treatment of the skin; and (iv) instructions for use.

[0216] The present invention provides, in particular, a technique for transdermal administration of large-cap drugs, such as botulinum toxin or antibody drugs, by incorporating one or more large-cap drugs into one or more emulsion compositions that are subsequently administered in combination with MSCs. The inventors have surprisingly found that the transdermal penetration and bioavailability of botulinum toxin or antibody drugs incorporated into nanoemulsion compositions are dramatically improved when used in combination with MSCs. The benefit of the present invention is the ability to administer such large-cap drugs intradermally while minimizing skin irritation or damage. The use of other drugs or processes in conjunction with the emulsion compositions and MSCs is not necessarily excluded, but is not required, in all embodiments of the present invention.

[0217] Therefore, the present invention provides a technique for administering large-capacity drugs by topically applying an excellent emulsion composition (e.g., a macroemulsion composition and / or a nanoemulsion composition) in combination with MSCs. In some embodiments, the large-capacity drug is botulinum toxin. In some embodiments, the botulinum emulsion composition is applied directly to the skin for absorption through the epidermal layer before the MSCs. In some embodiments, the botulinum emulsion composition is applied directly to the skin for absorption through the epidermal layer after the MSCs. In some embodiments, the botulinum emulsion composition is applied directly to the skin for absorption through the epidermal layer substantially simultaneously with the MSCs.

[0218] In some embodiments, botulinum emulsion compositions combined with MSCs can penetrate the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, without the use of penetration enhancers.

[0219] In some embodiments, the antibody-drug emulsion composition combined with MSCs can penetrate the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, without the use of penetration enhancers. In some embodiments, the main drug is an antibody drug. In some embodiments, the antibody-drug emulsion composition is applied directly to the skin for absorption through the epidermal layer before the MSCs. In some embodiments, the antibody-drug emulsion composition is applied directly to the skin for absorption through the epidermal layer after the MSCs. In some embodiments, the antibody-drug emulsion composition is applied directly to the skin for absorption through the epidermal layer substantially simultaneously with the MSCs. In some embodiments, the antibody-drug emulsion composition is applied directly to the skin for systemic absorption.

[0220] In some embodiments, antibody-drug emulsion compositions combined with MSCs can penetrate the uppermost layers of the skin, including the stratum corneum, dermal pores, and / or dermal glands, without the use of degrading agents, irritants, and / or abrasives.

[0221] Those skilled in the art will understand that the compositions for topical administration of the present invention may include cosmetic formulations such as skin softeners, nourishing lotion-type emulsions, cleansing lotions, cleansing creams, skin milks, emollient lotions, massage creams, emollient creams, cosmetic bases, face packs or face gels, cleansing formulations such as shampoos, conditioners, body washes, hair tonics or soaps, or skin compositions such as lotions, ointments, gels, creams, patches or sprays. In some embodiments, the compositions for topical administration are not formulated for administration to mucous membranes (for example, they are unsuitable for application to mucous membranes and / or are not formulated to deliver a suitable amount of a large drug to or beyond mucous membranes).

[0222] Those skilled in the art will understand that the units used herein relate to units that are biologically equivalent or bioactively equivalent to units defined by commercial manufacturers of botulinum toxin.

[0223] In some embodiments, the therapeutic effect of botulinum toxin administered according to the present invention may last as long as the effect of the injected solution persists. In some embodiments, the effect of such an injected solution may last for up to about 6-7 months. In some embodiments, the therapeutic effect of botulinum toxin administered according to the present invention may last for up to 6-7 months. In some embodiments, the use of a synthetic polymer carrier capable of holding the botulinum toxin so that the botulinum toxin is released gradually may extend the effect to up to about 5 years (U.S. Patent No. 6,312,708).

[0224] In some embodiments, the present invention provides topical formulations of botulinum toxin that avoid potential complications including, but not limited to, systemic toxicity or botulism. In some embodiments, the dosage of botulinum toxin (including botulinum toxin types A, B, C, D, E, F, or G, or botulinum toxin genetically engineered or chemically modified to have a longer or shorter duration of action than serotype A) may range from as low as about 1 unit to as high as about 20,000 units, with minimal risk of adverse side effects. The specific dosage may vary depending on the condition being treated and the treatment regimen being used. For example, treatment of subcutaneous hyperactive muscle may require a high transdermal dosage of botulinum toxin (e.g., 1,000 to 20,000 units). In contrast, treatment of neurogenic inflammation or hyperactive sweat glands may require a relatively low transdermal dosage of botulinum toxin (e.g., about 1 to 1,000 units).

[0225] Some embodiments of the present invention envision pharmaceutical compositions containing stabilized botulinum toxin for transdermal delivery to human patients. The botulinum toxin may be selected from the group consisting of botulinum toxin types A, B, C1, D, E, F, and G, isolated and / or purified (i.e., about 150 kDa) botulinum toxin, and naturally occurring or recombinantly produced botulinum toxin. In some embodiments, the composition may contain about 1 to about 20,000 units of botulinum toxin, and the composition may contain an amount of botulinum toxin sufficient to achieve a therapeutic effect lasting from 1 month to 5 years.

[0226] In some embodiments, the present invention provides topical formulations of botulinum toxin (e.g., botulinum emulsion compositions) that enable penetration through the target skin without significant penetration through the blood vessels. For example, in some embodiments of the present invention, less than about 25%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, or less than 1% of the botulinum toxin present in the pharmaceutical composition penetrates into the blood vessels upon application of the topical and / or transdermal preparations of the present invention.

[0227] In some embodiments, the present invention provides a topical formulation of an antibody drug (e.g., an antibody drug emulsion composition) that allows the antibody drug to penetrate through the target skin without penetrating in significant amounts through the blood vessels. For example, in some embodiments of the present invention, less than about 25%, or even less than 5%, of the antibody drug present in the pharmaceutical composition penetrates into the blood vessels upon application of the topical and / or transdermal preparation of the present invention.

[0228] In some embodiments, the present invention provides a topical formulation of an antibody drug (e.g., an antibody drug emulsion composition) that enables the antibody drug to penetrate through the skin of a target and penetrate through the blood vessels in a significant amount. In some embodiments, the present invention provides a topical formulation of an antibody drug (e.g., an antibody drug emulsion composition) that enables the antibody drug to penetrate through the skin of a target and penetrate through the blood vessels in a therapeutically effective amount. For example, in some embodiments of the present invention, about 25%, 50%, 75%, 90%, or more than 95% of the antibody drug present in the pharmaceutical composition penetrates into the blood vessels upon application of the topical and / or transdermal preparation of the present invention. In some embodiments, the present invention provides a topical formulation of an antibody drug (e.g., an antibody drug emulsion composition) that enables the antibody drug to exert a systemic effect on the target.

[0229] Those skilled in the art will understand that the compositions of the present invention, which achieve transdermal administration of botulinum toxin or antibody drugs, can be incorporated into devices such as patches. [Examples]

[0230] Example Example 1: Effect of microneedle skin preparation (MSC) pretreatment on the bioavailability of botulinum toxin. A single-dose local study was conducted to investigate the bioavailability of botulinum toxin after topical administration of botulinum nanoemulsion. The study was designed to test whether microneedle skin conditioning pretreatment (i.e., MSC performed before any administration of botulinum treatment as described herein) significantly enhances the bioavailability of botulinum.

[0231] The study included two test groups, each consisting of eight rats. Each group received a single topical treatment with a fixed volume of botulinum nanoemulsion at a fixed concentration to the skin covering the biceps femoris, gastrocnemius, and tibialis anterior muscles of the right hind limb. The administration of the topical preparation to the skin took approximately 10 minutes, at which point it was completely absorbed into the skin. The effect of such treatment was measured using a scale that measures limb paralysis, a known effect of botulinum administration to that limb. The scale is based on the finger abduction score (DAS score), a four-point scale developed by Aoki (2001, Toxicon, 39:1815; and 2002, Toxicon, 40:923), which can be readily assessed in treated rats. A score of 0 indicates no paralysis, and a score of 4 indicates complete paralysis. It is also known that doses of botulinum administered at concentrations sufficient to induce paralysis can be continued to induce death in animals. Therefore, the mortality rates were also compared between the two treatment groups.

[0232] As shown in Table 5, Group 1 did not undergo pre-treatment of the skin with MN. Group 2 was treated with five skin impressions using a microneedle array with 1400 mm needles covering each region of the biceps femoris, gastrocnemius, and tibialis anterior muscles of the right hind limb (i.e., skin pre-treatment was performed). Local treatment with botulinum nanoemulsion was thoroughly rubbed into the skin with gloved fingers until the treatment was no longer observable or palpable in the skin. [Table 5]

[0233] As shown in Figure 1, one day after treatment, animals treated with MSCs had approximately twice the DAS score of the control group that did not undergo such prior adjustment, establishing that MSCs unexpectedly increase the bioavailability of topical large-scale drug nanoemulsions containing botulinum toxin. Similarly, as shown in Figure 2, the mortality rate (100%) in animals treated with MSCs was approximately twice as high 40 hours after treatment compared to the mortality rate (50%) in the control group that did not undergo such prior adjustment, also establishing that MSCs unexpectedly increase the bioavailability of topical large-scale drug nanoemulsions containing botulinum toxin.

[0234] In short, the study results confirmed that MSCs significantly enhance the bioavailability of botulinum.

[0235] Example 2: Effect of MSC pre-treatment on the bioavailability of botulinum toxin in humans: Effect on sweat reduction This study will conduct a single-dose local study on the bioavailability of botulinum toxin after local administration of a topical botulinum nanoemulsion formulation in humans. The study is designed to test whether microneedle skin pre-treatment significantly enhanced botulinum bioavailability in humans by measuring the reduction in skin sweat after local treatment with the botulinum nanoemulsion formulation.

[0236] The study involves one subject. Two spots are selected on the abdomen, each approximately 2 cm square, and 5 cm apart, and marked with a marker. Each spot is treated once locally with a fixed volume of botulinum nanoemulsion preparation at a fixed concentration of botulinum. The administration of the topical preparation to the skin takes approximately 5 minutes, at which point the preparation is completely absorbed into the skin. The first spot is the control site and was not pre-prepared with a microneedle array. The second spot is the intervention site and was pre-prepared with three impressions using a 1400 micrometer length microneedle array before application of the botulinum preparation.

[0237] The expected effect of such treatment is a reduction in sweating at the site of botulinum nanoemulsion treatment. The amount of sweat at the treatment site is measured by two methods: 1) an evaporometer test (an instrument used to measure the rate of water evaporation from the skin is used to detect the rate of sweating, so that more evaporation is detected along with increased sweating); or 2) a starch-iodine test (a subject has povidine applied to the treatment site; it is dried; corn starch is sprinkled on the treatment site; if the subject's sweat enters the white corn starch, it turns purple; if the subject does not sweat, it remains white; this is called the starch-iodine test). In either sweat detection method, the subject is placed under a heat lamp to induce sweating, and then the sweat detection method is used.

[0238] Sweat detection methods were used at baseline before botulinum nanoemulsion treatment; and at 2 weeks and 4 weeks post-treatment. The study found that at baseline, the average sweat volume detected by either the evaporometer test or the starch-iodine test was approximately the same at the control and intervention sites. At 2 and 4 weeks post-treatment, the average sweat volume detected at the control site by either the evaporometer test or the starch-iodine test was higher than that detected at the intervention site during these post-treatment weeks.

[0239] This study establishes that microneedle pre-treatment unexpectedly increases the bioavailability of large topical drug nanoemulsions containing botulinum toxin.

[0240] Example 3: Effect of MSC pre-treatment on the bioavailability of botulinum toxin in humans: Effect on sweat reduction This study will conduct a single-dose local study on the bioavailability of botulinum toxin after local administration of a topical botulinum nanoemulsion formulation in humans. The study is designed to test whether microneedle skin pre-treatment significantly enhanced botulinum bioavailability in humans by measuring the reduction in skin sweat after local treatment with the botulinum nanoemulsion formulation.

[0241] The study included 12 subjects. Two spots were selected on each subject's back, each approximately 2 cm square, and 5 cm apart, and marked with a marker. Each spot was treated once locally with a fixed volume of botulinum nanoemulsion preparation at a fixed concentration of botulinum. The administration of the topical preparation to the skin took approximately 5 minutes, at which point the preparation was completely absorbed into the skin. The first spot was the control site and was not pre-prepared with a microneedle array. The second spot was the intervention site and was pre-prepared with three impressions using a 1400 micrometer length microneedle array before application of the botulinum preparation.

[0242] The expected effect of such treatment is a reduction in sweating at the site of botulinum nanoemulsion treatment. The amount of sweat at the treatment site is measured by two methods: 1) an evaporometer test (an instrument used to measure the rate of water evaporation from the skin is used to detect the rate of sweating, so that more evaporation is detected along with increased sweating); or 2) a starch-iodine test (a subject has povidine applied to the treatment site; it is dried; corn starch is sprinkled on the treatment site; if the subject's sweat enters the white corn starch, it turns purple; if the subject does not sweat, it remains white; this is called the starch-iodine test). For either sweat detection method, the subject is placed in a sauna to induce sweating, and then the sweat detection method is used.

[0243] Sweat detection methods were used at baseline before botulinum nanoemulsion treatment; and at 2 weeks and 4 weeks post-treatment. The study found that at baseline, the average sweat volume detected by either the evaporometer test or the starch-iodine test was approximately the same at the control and intervention sites. At 2 and 4 weeks post-treatment, the average sweat volume detected at the control site by either the evaporometer test or the starch-iodine test was higher than that detected at the intervention site during these post-treatment weeks.

[0244] This study establishes that microneedle pre-treatment unexpectedly increases the bioavailability of large topical drug nanoemulsions containing botulinum toxin.

[0245] Example 4: Effect of MSC pre-treatment on the bioavailability of botulinum toxin in humans: Effect on sweat reduction This study will conduct a single-dose local study on the bioavailability of botulinum toxin after local administration of a topical botulinum nanoemulsion formulation in humans. The study is designed to test whether microneedle skin conditioning significantly enhanced the bioavailability of botulinum in humans by measuring the reduction in skin sweat after local treatment with the botulinum nanoemulsion formulation.

[0246] The study involved one subject. Two spots were selected on the abdomen, each approximately 3 cm square and 5 cm apart, and marked with a marker. Each spot was treated once topically with a fixed volume of botulinum nanoemulsion. The topical preparation took approximately 5 minutes to be administered to the skin, at which point it was completely absorbed into the skin. The first spot was the control site, and was not pre-prepared with a microneedle array before application of the botulinum preparation. The second spot was the intervention site, and was pre-prepared with three impressions using a 1400 micrometer microneedle array before application of the botulinum preparation. The concentration of botulinum preparation administered to the intervention site was half that of the control site.

[0247] The expected effect of such treatment is a reduction in sweating at the site of botulinum nanoemulsion treatment. The amount of sweat at the treatment site is measured by two methods: 1) an evaporometer test (an instrument used to measure the rate of water evaporation from the skin is used to detect the rate of sweating, so that more evaporation is detected along with increased sweating); or 2) a starch-iodine test (a subject has povidine applied to the treatment site; it is dried; corn starch is sprinkled on the treatment site; if the subject's sweat enters the white corn starch, it turns purple; if the subject does not sweat, it remains white; this is called the starch-iodine test). In either sweat detection method, the subject is placed under a heat lamp to induce sweating, and then the sweat detection method is used.

[0248] Sweat detection methods were used at baseline before botulinum nanoemulsion treatment; and at 2 weeks and 4 weeks post-treatment. The study found that at baseline, the mean sweat volume detected by either the evaporative sweat test or the starch-iodine test was nearly equal at the control and intervention sites. At 2 and 4 weeks post-treatment, the mean sweat volume detected by either the evaporative sweat test or the starch-iodine test at the control site was equivalent to that at the intervention site, despite the fact that the concentration of the botulinum preparation administered to the intervention site was half that of the control site. This study establishes that microneedle pre-adjustment unexpectedly increases the bioavailability of topical large drug nanoemulsions containing botulinum toxin.

[0249] Example 5: Effect of microneedle treatment (MN) skin pre-conditioning on the bioavailability of botulinum toxin in humans: Effect on reducing sweat and wrinkles. This study will conduct a single-dose local study on the bioavailability of botulinum toxin after local administration of a topical botulinum nanoemulsion formulation in humans. The study is designed to test whether microneedle skin pre-treatment significantly enhanced the bioavailability of botulinum in humans by measuring the reduction in skin sweat and wrinkles after local treatment with the botulinum nanoemulsion formulation.

[0250] The study included one subject with severe frontal (or horizontal) wrinkles on the forehead. Two spots were selected on the subject's forehead, each approximately 2 cm square and 5 cm apart, and marked with a marker. Each spot was treated once topically with a fixed volume of botulinum nanoemulsion at a fixed concentration of botulinum. The administration of the topical preparation to the skin took approximately 5 minutes, at which point the preparation was completely absorbed into the skin. The first spot was the control site and was not pre-prepared with a microneedle array. The second spot was the intervention site and was pre-prepared with three impressions of a 1400 micrometer length microneedle array before application of the botulinum preparation.

[0251] The expected effect of such treatment is a reduction in sweating at the site of botulinum nanoemulsion treatment. The amount of sweat at the treatment site is measured by two methods: 1) an evaporometer test (an instrument used to measure the rate of water evaporation from the skin is used to detect the rate of sweating, so that more evaporation is detected along with increased sweating); or 2) a starch-iodine test (a subject has povidine applied to the treatment site; it is dried; corn starch is sprinkled on the treatment site; if the subject's sweat enters the white corn starch, it turns purple; if the subject does not sweat, it remains white; this is called the starch-iodine test). For either sweat detection method, the subject is placed in a sauna to induce sweating, and then the method is used.

[0252] The expected effect of botulinum nanoemulsion treatment is a reduction in frontal wrinkles at the treatment site. The severity of wrinkles is measured using a 4-point wrinkle scale: 0 = none, 1 = mild, 2 = moderate, 3 = severe.

[0253] Sweat detection methods were used at baseline before botulinum nanoemulsion treatment; and at 2 weeks and 4 weeks post-treatment. The study found that at baseline, the mean sweat volume detected by either the evaporometer test or the starch-iodine test was approximately equal in the control and intervention sites. At 2 and 4 weeks post-treatment, the mean sweat volume detected by either the evaporometer test or the starch-iodine test in the control site was higher than that detected in the intervention site during these post-treatment weeks. The study found that at baseline, the mean severity of frontal wrinkles, measured by the wrinkle scale, was approximately equal in the control and intervention sites. At 2 and 4 weeks post-treatment, the mean severity of frontal wrinkles, measured by the wrinkle scale, in the control site was higher than that detected in the intervention site during these post-treatment weeks.

[0254] This study establishes that microneedle pre-treatment unexpectedly increases the bioavailability of large topical drug nanoemulsions containing botulinum toxin.

[0255] Example 6: Effect of microneedle treatment (MN) skin pre-conditioning on the bioavailability of botulinum toxin in humans: Effect on reducing sweat in subjects with hyperhidrosis This study will conduct a single-dose local study on the bioavailability of botulinum toxin after local administration of a topical botulinum nanoemulsion formulation in humans. The study is designed to test whether microneedle skin pre-treatment significantly enhanced botulinum bioavailability in humans by measuring the reduction in skin sweat after local treatment with the botulinum nanoemulsion formulation.

[0256] The study included two treatment groups of 20 subjects each with axillary hyperhidrosis, a pathological condition characterized by excessive sweating in the armpits: Group 1 was the control group, receiving botulinum nanoemulsion applied to each subject's armpit; Group 2 was the intervention group, pre-preparing each part of the armpit skin with three impressions using a 1400-micrometer microneedle array before application of the botulinum nanoemulsion formulation. Each subject in Groups 1 and 2 received one topical treatment with a fixed volume of botulinum nanoemulsion formulation at a fixed concentration of botulinum. The topical formulation took approximately 5 minutes to be fully absorbed into the skin.

[0257] The expected effect of such treatment is a reduction in sweating at the site of botulinum nanoemulsion treatment, which is the armpit. The amount of sweat at the treatment site is measured by gravimetric sweating (GS test): the subject's armpit is dried with a paper towel; filter paper is weighed; the filter paper is applied to the armpit for 5 minutes; then weighed again; the excess weight after re-weighing the paper is the weight of sweat produced by the subject in 5 minutes. The severity of the subject's hyperhidrosis is measured by the subject using the Hyperhidrosis Sweating Severity Scale (HDSS), a 4-point scale assessed by the subject: 0 = none, 1 = mild, 2 = moderate, 3 = severe.

[0258] The GS test and HDSS were used at baseline before botulinum nanoemulsion treatment; and at 2 weeks and 4 weeks post-treatment. The study found that at baseline, the mean sweat volume detected by the GS test or disease severity measured by HDSS was approximately equal across groups 1 and 2. At 2 and 4 weeks post-treatment, the mean sweat volume or disease severity detected in group 1 was greater than that detected in group 2 during those post-treatment weeks.

[0259] This study establishes that microneedle pre-treatment unexpectedly increases the bioavailability of large topical drug nanoemulsions containing botulinum toxin.

[0260] Example 7: Effect of microneedle treatment (MN) skin pre-treatment on the bioavailability of botulinum toxin in humans: Effect on reducing crow's feet wrinkles. A single-dose local study was conducted to evaluate the bioavailability of botulinum toxin after topical administration of a topical botulinum nanoemulsion formulation in humans. The study was designed to test whether microneedle skin pre-treatment significantly enhanced botulinum bioavailability in humans by measuring wrinkle reduction in the skin after topical treatment with the botulinum nanoemulsion formulation.

[0261] The study included one subject who had severe crow's feet wrinkles on the side of her eye. Botulinum nanoemulsion was applied to the subject's crow's feet wrinkles. The dose of botulinum applied to the skin was approximately 15% of the effective dose when botulinum nanoemulsion was applied without microneedle skin pre-treatment. The effective dose was defined as the dose that would produce at least a 2-point improvement in the appearance of the wrinkles when the subject was contracting the muscles that cause crow's feet wrinkles, as measured by a 5-point wrinkle rating scale. The subject was pre-treated with two impressions of a 1400-micrometer microneedle array on each area of ​​skin where crow's feet wrinkles were located before application of the botulinum nanoemulsion formulation to one side of the face, and with two impressions of an 800-micrometer microneedle array on each area of ​​skin where crow's feet wrinkles were located before application of the botulinum emulsion formulation to the other side of the face. The topical formulation was administered to the skin in approximately 5 minutes, at which point it was completely absorbed into the skin.

[0262] The expected effect of botulinum nanoemulsion treatment is a reduction in crow's feet wrinkles at the treatment site. Wrinkle severity was measured using a 5-point wrinkle scale: 0=none, 1=minimal, 2=mild, 3=moderate, 4=severe.

[0263] The wrinkle scale was used at baseline before botulinum nanoemulsion treatment, 2 weeks after treatment, and 4 weeks after treatment. In the study, at baseline, the subjects were found to have severe wrinkles with a score of 4 on a 5-point wrinkle scale, as evaluated by the wrinkle scale. Two weeks after treatment, the average severity of wrinkles decreased by 1 point on the wrinkle scale to a score of 3 (moderate) on each side of the face. Four weeks after treatment, the average severity of wrinkles decreased by 2 points on the wrinkle scale to a score of 2 (mild) on each side of the face.

[0264] This study established that microneedle preconditioning unexpectedly increased the bioavailability of a topical large drug nanoemulsion containing botulinum toxin.

[0265] Example 8: Effect of microneedle treatment (MN) skin preconditioning on the bioavailability of botulinum toxin in humans: Effect on reduction of crow's foot wrinkles A single-dose topical study of the bioavailability of botulinum toxin after topical administration of a topical botulinum nanoemulsion formulation in humans is conducted. The study is designed to test whether microneedle skin preconditioning significantly enhances the bioavailability of botulinum in humans by measuring wrinkle reduction in the skin after topical treatment with the botulinum nanoemulsion formulation.

[0266] The study includes two treatment groups of 20 subjects, each subject having severe crow's foot wrinkles on their side of the eye: Group 1 is the control group and has the botulinum nanoemulsion applied to the crow's foot wrinkles of each subject; Group 2 is the intervention group and is preconditioned with three impressions of a 1400 micrometer long microneedle array on each part of the skin where the crow's foot wrinkles are located before application of the botulinum nanoemulsion formulation. Each subject in Groups 1 and 2 is topically treated once with a fixed volume of the botulinum nanoemulsion formulation at a fixed concentration of botulinum. The topical administration of the formulation to the skin takes about 5 minutes, at which point the topical formulation is completely absorbed into the skin.

[0267] The expected effect of botulinum nanoemulsion treatment is a reduction in crow's feet wrinkles at the treatment site. The severity of wrinkles is measured using a 4-point wrinkle scale: 0 = none, 1 = mild, 2 = moderate, 3 = severe.

[0268] The wrinkle scale was used at baseline before botulinum nanoemulsion treatment, and at 2 weeks and 4 weeks post-treatment. The study found that at baseline, the mean severity of wrinkles detected by the wrinkle scale was nearly equal across groups 1 and 2. At 2 and 4 weeks post-treatment, the mean severity of wrinkles in group 1 was higher than that detected in group 2 at those post-treatment weeks.

[0269] This study establishes that microneedle pre-treatment unexpectedly increases the bioavailability of large topical drug nanoemulsions containing botulinum toxin.

[0270] Example 9: Effect of microneedle treatment (MN) skin pre-treatment on the bioavailability of botulinum toxin in humans: Effect of dose change on the reduction of crow's foot wrinkles. This study will conduct a single-dose local study on the bioavailability of botulinum toxin after local administration of a topical botulinum nanoemulsion formulation in humans. The study is designed to test whether microneedle skin pre-treatment significantly enhanced the bioavailability of botulinum in humans by measuring wrinkle reduction in the skin after local treatment with the botulinum nanoemulsion formulation.

[0271] The study included two treatment groups of 20 subjects, each subject having severe crow's feet wrinkles: Group 1 was the control group, with botulinum nanoemulsion applied to each subject's crow's feet wrinkles; Group 2 was the intervention group, pre-treated by three impressions of a 1400-micrometer microneedle array on each part of the skin where the crow's feet wrinkles were located before application of the botulinum nanoemulsion formulation. Each subject was treated once topically with a fixed volume of botulinum nanoemulsion formulation at a fixed concentration of botulinum, except that Group 1 treatment used twice the botulinum concentration as Group 2 treatment. The topical formulation took approximately 5 minutes to be fully absorbed into the skin.

[0272] The expected effect of botulinum nanoemulsion treatment is a reduction in crow's feet wrinkles at the treatment site. The severity of wrinkles is measured using a 4-point wrinkle scale: 0 = none, 1 = mild, 2 = moderate, 3 = severe.

[0273] The wrinkle scale was used at baseline before botulinum nanoemulsion treatment, and at 2 weeks and 4 weeks post-treatment. The study found that at baseline, the mean severity of wrinkles detected by the wrinkle scale was nearly equal across groups 1 and 2. At 2 and 4 weeks post-treatment, the mean severity of wrinkles in groups 1 and 2 decreased by nearly the same amount compared to baseline, even though group 1 was treated with twice the concentration of group 2.

[0274] This study establishes that microneedle pre-treatment unexpectedly increases the bioavailability of large topical drug nanoemulsions containing botulinum toxin, allowing lower doses of botulinum to be used to achieve equivalent therapeutic effects compared to patients who did not undergo microneedle skin pre-treatment.

[0275] Example 10: Effect of microneedle treatment (MN) skin pre-treatment on the bioavailability of botulinum toxin in humans: Effect of dose change on the reduction of crow's foot wrinkles. This study will conduct a single-dose local study on the bioavailability of botulinum toxin after local administration of a topical botulinum macroemulsion formulation in humans. The study is designed to test whether microneedle skin pre-treatment significantly enhanced the bioavailability of botulinum in humans by measuring wrinkle reduction in the skin after local treatment with the botulinum macroemulsion formulation.

[0276] The study included two treatment groups of 20 subjects, each subject having severe crow's feet wrinkles: Group 1 was the control group, with botulinum macroemulsion applied to each subject's crow's feet wrinkles; Group 2 was the intervention group, pre-prepared by three impressions of a 1400-micrometer microneedle array on each part of the skin where the crow's feet wrinkles were located, before application of the botulinum macroemulsion preparation. Each subject received a single topical treatment with a fixed volume of botulinum nanoemulsion preparation at a fixed concentration of botulinum. The topical preparation took approximately 5 minutes to be administered to the skin, at which point it was completely absorbed into the skin.

[0277] The expected effect of botulinum nanoemulsion treatment is a reduction in crow's feet wrinkles at the treatment site. The severity of wrinkles is measured using a 4-point wrinkle scale: 0 = none, 1 = mild, 2 = moderate, 3 = severe.

[0278] The wrinkle scale was used at baseline before botulinum nanoemulsion treatment, and at 2 weeks and 4 weeks post-treatment. The study found that at baseline, the mean severity of wrinkles detected by the wrinkle scale was approximately the same across groups 1 and 2. At 2 and 4 weeks post-treatment, the mean severity of wrinkles in group 1 was greater than that in group 2.

[0279] This study establishes that microneedle pre-treatment unexpectedly increases the bioavailability of topical large drug macroemulsions containing botulinum toxin compared to patients who did not undergo microneedle dermal pre-treatment.

[0280] References Alkilani, A.Z., et al., “Transdermal drug delivery: innovative pharmaceutical developments based on disruption of the barrier properties of the stratum corneum.” Pharmaceutics. 7:438 - 470(2015). Gomaa, Y., et al, “Effect of microneedle treatment on the skin permeation of a nanoencapsulated dye.” J Pharm Pharmacol. 2012 November ;64(11):1592 - 1602. Guo, L., et al., “Enhanced transcutaneous immunization via dissolving microneedle array loaded with liposome encapsulated antigen and adjuvant.” International Journal of Pharmaceutics. 447:22 - 30(2013) Hiraishi, Y., et al., “Development of a novel therapeutic approach using a retinoic acid - loaded microneedle patch for seborrheic keratosis treatment and safety study in humans.” Journal of Controlled Release. 171:93 - 103(2013). Ling, M. & Chen, M., “Dissolving polymer microneedle patches for rapid and efficient transdermal delivery of insulin to diabetic rats.” Acta Biomaterialia, 9, 8952 - 8961(2013). Prausnitz,M.R.& Langer,R.“Transdermal drug delivery,”Nat Biotechnol.26(11):1261-1268(2008). Qin,G.,et al.,“Simultaneous basal-bolus delivery of fast-acting insulin and its significance in diabetes management.” Nanomedicine:NBM,8:221-227(2012). Tang,H.,et al.,“Theoretical description of transdermal transport of hydrophilic permeants:Application to low-frequency sonophoresis.” Journal of Pharmaceutical Sciences.90(5):545-568(2001). Verbaan,F.J.,et al.,“Assembled microneedle arrays enhance the transport of compounds varying over a large range of molecular weight across human dermatomed skin.” Journal of Controlled Release.117:238-245(2007). Wu,X.,et al.,“Effects of pretreatment of needle puncture and sandpaper abrasion on the in vitro skin permeation of fluorescein isothiocyanate(FITC)-dextran.” International Journal of Pharmaceutics.316:102-108(2006). Zhang, S., et al., “Enhanced bioavailability of L-carnitine after painless intradermal delivery vs. oral administration in rats.” Pharm Res. 28:117-123 (2011). Zhang, S., et al., “Enhanced delivery of hydrophilic peptides in vitro by transdermal microneedle pretreatment.” Acta Pharmaceutica Sinica B. 4(1):100-104 (2014).

[0281] Equivalents One of ordinary skill in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. The scope of the present invention is not intended to be limited to the above description, but rather is as set forth in the following claims. The present invention provides the following items. (Item 1) A method comprising applying, in combination with microneedle skin conditioning (MSC) of a site, an emulsion composition containing a large drug having a molecular weight of 100,000 KDa or more to the site. (Item 2) The method according to item 1, wherein the composition containing the large drug contains a nanoemulsion containing the large drug. (Item 3) The method according to item 1, wherein the composition containing the large drug contains a macroemulsion containing the large drug. (Item 4) The method according to any one of items 1 to 3, further comprising administration of a non-irritating penetration enhancer. (Item 5) The method according to item 4, wherein the non-irritating penetration enhancer is selected from a carrier peptide and a copetide. (Item 6) The non-irritating penetration promoter is a cationic peptide and sequence RKKRRQRRRG-(K) 15 - A method according to any one of items 4 to 5, selected from positively charged carriers having GRKKRRQRRR. (Item 7) The method according to any one of items 1 to 6, wherein the MSC of the site is performed before the composition containing the large drug is applied to the site. (Item 8) The method according to any one of items 1 to 6, wherein the MSC in the said area is performed after the composition containing the large drug is applied to the said area. (Item 9) The method according to any one of items 1 to 6, wherein the composition comprising the MSC and the large drug at the site is applied to the site substantially simultaneously. (Item 10) The method according to any one of items 1 to 9, wherein the large drug is botulinum toxin. (Item 11) The method according to item 10, further comprising delivering botulinum toxin together with a biological activator. (Item 12) The method according to item 11, wherein the biological activator is selected from steroids, retinoids, anesthetics, fillers, silicones, and / or collagen. (Item 13) The method according to item 12, wherein the biological activator is selected from hydrocortisone, retin-A, and / or lidocaine. (Item 14) The method according to any one of items 1 to 9, wherein the aforementioned large drug is an antibody drug. (Item 15) The method according to item 14, wherein the antibody drug is selected from anti-TNFα antibody, anti-CD2 antibody, anti-CD4 antibody, anti-IL-12 antibody, anti-IL-17 antibody, anti-IL-22 antibody, and anti-IL-23 antibody. (Item 16) The method according to any one of items 14 to 15, wherein the antibody drug is selected from antibodies having an epitope-binding element found in one or more of the following: infliximab, adalimumab, golimumab, etanercept, etanercept-szzs, certolizumab pegol, cipirizumab, zanolimumab, briakinumab, secukinumab, brodalumab, fezakinumab, ustekinumab and / or guselkumab. (Item 17) The method according to any one of items 14 to 16, further comprising delivering the antibody drug together with a biological activator. (Item 18) The method according to any one of items 14 to 17, further comprising delivering the antibody drug together with a non-irritating penetration promoter. (Item 19) The method according to item 18, wherein the non-irritating penetration promoter is selected from copolymers and carrier peptides. (Item 20) The method according to any one of items 1 to 19, wherein the MSC of the said site is achieved with a device including a plurality of needles. (Item 21) The method according to item 20, wherein the device is a patch, roller, stamp, or pen. (Item 22) The method according to any one of items 1 to 21, wherein the area is the skin surface covering the target muscle or muscle group. (Item 23) The method according to any one of items 1 to 22, wherein the said area is a skin surface containing sweat glands. (Item 24) The method according to any one of items 1 to 23, wherein the aforementioned area is a skin surface containing sebaceous glands. (Item 25) The method according to any one of items 1 to 24, wherein the said area is a skin surface containing a hair follicle. (Item 26) The method according to any one of items 20 to 25, wherein the needle is of sufficient length to protrude through the stratum corneum of the skin. (Item 27) The method according to any one of items 20 to 26, wherein the needle is of insufficient length to reach the nerves in the dermis of the skin. (Item 28) The method according to any one of items 20 to 27, wherein the needle has a length of about 10 to about 4000 μm. (Item 29) The method according to any one of items 20 to 28, wherein the needle has a length of approximately 100 μm or more. (Item 30) The method according to any one of items 20 to 28, wherein the needle has a length of approximately 25 μm or more. (Item 31) The method according to any one of items 20 to 28, wherein the needle has a length of approximately 300 μm, approximately 500 μm, approximately 800 μm, approximately 1000 μm, approximately 1500 μm, approximately 2000 μm, or approximately 4000 μm or more. (Item 32) The method according to any one of items 20 to 31, wherein the needle is made of a biocompatible material. (Item 33) The method according to any one of items 20 to 31, wherein the needle is made of metal. (Item 34) The method according to any one of items 1 to 33, wherein the MSC includes administering 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 MN or MN array impressions. (Item 35) The method according to item 34, wherein the MN array is rotated between one or more impressions. (Item 36) The method according to item 34, wherein the MN array is not rotated between one or more impressions. (Item 37) The method described in any one of items 34-36, wherein the aforementioned impression is taken on approximately the same area. (Item 38) The method described in any one of items 34 to 36, wherein the aforementioned impression is performed on the overlapping area. (Item 39) The method described in any one of items 34-36, wherein the aforementioned impression is performed on a different part of the body. (Item 40) The method according to any one of items 34 to 39, wherein the MN array is in the form of a stamp or a roller. (Item 41) The method according to item 40, wherein the impression is made by stamping or by rotating a roller. (Item 42) The method according to any one of items 1 to 41, wherein the large-capacity drug penetrates the skin within approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 minutes after administration. (Item 43) The method according to any one of items 1 to 41, wherein the large drug penetrates the skin within approximately 5 to 60 minutes, 5 to 12 minutes, 5 to 15 minutes, or 15 to 30 minutes after administration. (Item 44) The method according to any one of items 1 to 41, wherein the large-capacity drug penetrates the skin within approximately 1, 2, 3, 4, 5, or 6 hours after administration. (Item 45) The method according to any one of items 20 to 32, wherein the needle is composed of a soluble polymer. (Item 46) The method according to any one of items 1 to 45, wherein the administration comprises administering the composition containing a major drug at a smaller dose compared to a reference local treatment regimen in which a microneedle is not used. (Item 47) The method according to any one of items 1 to 46, comprising administering the composition containing multiple doses of a large-cap drug over time. (Item 48) The method according to item 47, wherein the administration involves administering the composition containing a smaller dose of a large-capacity drug over a period of time compared to a reference treatment regimen in which microneedles are not used, thereby producing an equivalent therapeutic effect. (Item 49) The method according to item 47 or item 48, wherein each dose of the composition containing a large-capacitor agent is separated by a specific period of time. (Item 50) The method according to item 49, wherein the aforementioned specific period is longer than the specific period for administering a reference treatment regimen in which microneedles are not used. (Item 51) The method according to any one of items 46 to 50, wherein the reference treatment regimen includes administering the composition containing a major drug without the presence of MSCs. (Item 52) A method for treating a skin disorder, comprising the method described in any one of items 1 through 51. (Item 53) The method according to item 52, wherein the skin disorder is selected from acne, excessive sweating, body odor, hyperhidrosis, bromhidrosis, chromatohidrosis, rosacea, alopecia, psoriasis, actinic keratosis, eczematous dermatitis, sebum production disorder, burns, lupus erythematosus, hyperpigmentation disorder, hypopigmentation disorder, skin cancer, skin infection, facial wrinkles, unattractive facial expressions, neck, multifunctional facial muscles, hypercontracted facial muscles, platysma girdles, and / or combinations thereof. (Item 54) Unwanted sweating, body odor, hyperhidrosis, bromhidrosis, chromatohidrosis, alopecia, Raynaud's phenomenon, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, lupus erythematosus, systemic lupus erythematosus, lupus discoid, drug-induced lupus, neonatal lupus, Crohn's disease, inflammatory bowel disease, ulcerative colitis, lung disease, asthma, chronic obstructive airway disorder, amyloidosis, systemic amyloidosis, cutaneous amyloidosis, cancer, skin cancer, hematological cancer, breast cancer, colon cancer, lung cancer, benign prostatic hyperplasia, dyslipidemia, hypercholesterolemia Methods for treating or preventing disorders selected from infectious diseases, C. difficile infection, Staphylococcus infection, dystonia, headache, pain, pain associated with arthritis, pain associated with rheumatoid arthritis, pain associated with psoriatic arthritis, pain associated with osteoarthritis, specified ophthalmic conditions, specified urinary tract conditions, neuromuscular disorders, conditions including muscle spasms and / or contractures, strabismus, hemifacial spasm, tremor, spasticity such as that resulting from multiple sclerosis, posterior orbital muscle, neurological conditions, Alzheimer's disease, Parkinson's disease, or stroke. (Item 55) The method according to any one of items 1 to 54, wherein the composition containing a large drug is formulated as a lotion, cream, powder, ointment, liniment, gel, or drop. (Item 56) A patch comprising a composition containing a large drug molecule and a plurality of microneedles. (Item 57) The patch according to item 56, wherein the composition containing the large drug compound comprises a nanoemulsion. (Item 58) The patch according to item 56, wherein the composition containing the large drug compound comprises a macroemulsion. (Item 59) The patch according to any one of items 56 to 58, wherein the needle has a length sufficient to protrude through the stratum corneum of the skin. (Item 60) The patch according to any one of items 57 to 59, wherein the needle is of insufficient length to reach the nerves in the dermis of the skin. (Item 61) The patch according to any one of items 57 to 60, wherein the needle has a length of approximately 10 to approximately 1000 μm. (Item 62) The patch described in any one of items 57 to 61, wherein the needle has a length of approximately 100 μm or more. (Item 63) The patch described in any one of items 57 to 62, wherein the needle has a length of approximately 300 μm or more. (Item 64) The aforementioned needle is made of a biocompatible material, and the patch is as described in any one of items 57 to 63. (Item 65) The aforementioned needle is made of metal, and the patch is as described in any one of items 57-64. (Item 66) The aforementioned needle is made of a soluble polymer, and the patch is as described in any one of items 57 to 64. (Item 67) The aforementioned large drug has a molecular weight of 100 kDa or more, and is a patch according to any one of items 57 to 66. (Item 68) The patch described in any one of items 57-67, wherein the aforementioned large drug is botulinum toxin. (Item 69) The aforementioned large drug is an antibody drug, and the patch is one of the items in items 57 to 67. (Item 70) A kit comprising a composition containing a large-capacitor agent, and a patch for use in applying the composition containing the large-capacitor agent to a site. (Item 71) The kit according to item 70, wherein the composition containing the large-capacitor agent is incorporated into the patch. (Item 72) A kit comprising a composition containing a large drug and a device for adjusting the microneedle at a site. (Item 73) The kit according to any one of items 70 to 72, wherein the composition containing a large drug compound comprises a nanoemulsion. (Item 74) The kit according to any one of items 70 to 72, wherein the composition containing a large drug comprises a macroemulsion. (Item 75) The device is a kit according to any one of items 70 to 74, comprising a plurality of needles. (Item 76) The kit described in any one of items 70-75, wherein the device is a patch, roller, stamp, or pen. (Item 77) A kit as described in one of items 70-76, including the patch described in one of items 56-68. (Item 78) A kit according to any one of items 70 or 72-77, wherein the composition containing a large drug is formulated as a lotion, cream, powder, ointment, liniment, gel, or drop. The present invention provides, for example, the following items: (Item 1) A method comprising applying an emulsion composition containing a large drug having a molecular weight of 100,000 Da or more to a site in combination with microneedle skin conditioning (MSC) of the site. (Item 2) The method according to item 1, wherein the composition containing the large-capacitor compound comprises a nanoemulsion containing the large-capacitor compound. (Item 3) The method according to item 1, wherein the composition containing the large-capacitor compound comprises a macroemulsion containing the large-capacitor compound. (Item 4) The method according to any one of items 1 to 3, further comprising the administration of a non-irritating penetration promoter. (Item 5) The method according to item 4, wherein the non-irritating penetration promoter is selected from carrier peptides and copolymers. (Item 6) The non-irritating penetration promoter is a cationic peptide and sequence RKKRRQRRRG-(K) 15 - A method according to any one of items 4 to 5, selected from positively charged carriers having GRKKRRQRRR. (Item 7) The method according to any one of items 1 to 6, wherein the MSC of the site is performed before the composition containing the large drug is applied to the site. (Item 8) The method according to any one of items 1 to 6, wherein the MSC in the said area is performed after the composition containing the large drug is applied to the said area. (Item 9) The method according to any one of items 1 to 6, wherein the composition comprising the MSC and the large drug at the site is applied to the site substantially simultaneously. (Item 10) The method according to any one of items 1 to 9, wherein the large drug is botulinum toxin. (Item 11) The method according to item 10, further comprising delivering botulinum toxin together with a biological activator. (Item 12) The method according to item 11, wherein the biological activator is selected from steroids, retinoids, anesthetics, fillers, silicones, and / or collagen. (Item 13) The method according to item 12, wherein the biological activator is selected from hydrocortisone, retin-A, and / or lidocaine. (Item 14) The method according to any one of items 1 to 9, wherein the aforementioned large drug is an antibody drug. (Item 15) The method according to item 14, wherein the antibody drug is selected from anti-TNFα antibody, anti-CD2 antibody, anti-CD4 antibody, anti-IL-12 antibody, anti-IL-17 antibody, anti-IL-22 antibody, and anti-IL-23 antibody. (Item 16) The method according to any one of items 14 to 15, wherein the antibody drug is selected from antibodies having an epitope-binding element found in one or more of the following: infliximab, adalimumab, golimumab, etanercept, etanercept-szzs, certolizumab pegol, cyprizumab, zanolimumab, briakinumab, secukinumab, brodalumab, fezakinumab, ustekinumab, and / or guselkumab. (Item 17) The method according to any one of items 14 to 16, further comprising delivering the antibody drug together with a biological activator. (Item 18) The method according to any one of items 14 to 17, further comprising delivering the antibody drug together with a non-irritating penetration promoter. (Item 19) The method according to item 18, wherein the non-irritating penetration promoter is selected from copolymers and carrier peptides. (Item 20) The method according to any one of items 1 to 19, wherein the MSC of the said site is achieved with a device including a plurality of needles. (Item 21) The method according to item 20, wherein the device is a patch, roller, stamp, or pen. (Item 22) The method according to any one of items 1 to 21, wherein the area is the skin surface covering the target muscle or muscle group. (Item 23) The method according to any one of items 1 to 22, wherein the said area is a skin surface containing sweat glands. (Item 24) The method according to any one of items 1 to 23, wherein the aforementioned area is a skin surface containing sebaceous glands. (Item 25) The method according to any one of items 1 to 24, wherein the said area is a skin surface containing a hair follicle. (Item 26) The method according to any one of items 20 to 25, wherein the needle is of sufficient length to protrude through the stratum corneum of the skin. (Item 27) The method according to any one of items 20 to 26, wherein the needle is of insufficient length to reach the nerves in the dermis of the skin. (Item 28) The method according to any one of items 20 to 27, wherein the needle has a length of about 10 to about 4000 μm. (Item 29) The method according to any one of items 20 to 28, wherein the needle has a length of approximately 100 μm or more. (Item 30) The method according to any one of items 20 to 28, wherein the needle has a length of approximately 25 μm or more. (Item 31) The method according to any one of items 20 to 28, wherein the needle has a length of approximately 300 μm, approximately 500 μm, approximately 800 μm, approximately 1000 μm, approximately 1500 μm, approximately 2000 μm, or approximately 4000 μm or more. (Item 32) The method according to any one of items 20 to 31, wherein the needle is made of a biocompatible material. (Item 33) The method according to any one of items 20 to 31, wherein the needle is made of metal. (Item 34) The method according to any one of items 1 to 33, wherein the MSC includes administering 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 MN or MN array impressions. (Item 35) The method according to item 34, wherein the MN array is rotated between one or more impressions. (Item 36) The method according to item 34, wherein the MN array is not rotated between one or more impressions. (Item 37) The method described in any one of items 34-36, wherein the aforementioned impression is taken on approximately the same area. (Item 38) The method described in any one of items 34 to 36, wherein the aforementioned impression is performed on the overlapping area. (Item 39) The method described in any one of items 34-36, wherein the aforementioned impression is performed on a different part of the body. (Item 40) The method according to any one of items 34 to 39, wherein the MN array is in the form of a stamp or a roller. (Item 41) The method according to item 40, wherein the impression is made by stamping or by rotating a roller. (Item 42) The method according to any one of items 1 to 41, wherein the large-capacity drug penetrates the skin within approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 minutes after administration. (Item 43) The method according to any one of items 1 to 41, wherein the large drug penetrates the skin within approximately 5 to 60 minutes, 5 to 12 minutes, 5 to 15 minutes, or 15 to 30 minutes after administration. (Item 44) The method according to any one of items 1 to 41, wherein the large-capacity drug penetrates the skin within approximately 1, 2, 3, 4, 5, or 6 hours after administration. (Item 45) The method according to any one of items 20 to 32, wherein the needle is composed of a soluble polymer. (Item 46) The method according to any one of items 1 to 45, wherein the administration comprises administering the composition containing a major drug at a smaller dose compared to a reference local treatment regimen in which a microneedle is not used. (Item 47) The method according to any one of items 1 to 46, comprising administering the composition containing multiple doses of a large-cap drug over time. (Item 48) The method according to item 47, wherein the administration involves administering the composition containing a smaller dose of a large-capacity drug over a period of time compared to a reference treatment regimen in which microneedles are not used, thereby producing an equivalent therapeutic effect. (Item 49) The method according to item 47 or item 48, wherein each dose of the composition containing a large-capacitor agent is separated by a specific period of time. (Item 50) The method according to item 49, wherein the aforementioned specific period is longer than the specific period for administering a reference treatment regimen in which microneedles are not used. (Item 51) The method according to any one of items 46 to 50, wherein the reference treatment regimen includes administering the composition containing a major drug without the presence of MSCs. (Item 52) A method for treating a skin disorder, comprising the method described in any one of items 1 through 51. (Item 53) The method according to item 52, wherein the skin disorder is selected from acne, excessive sweating, body odor, hyperhidrosis, bromhidrosis, chromatohidrosis, rosacea, alopecia, psoriasis, actinic keratosis, eczematous dermatitis, sebum production disorder, burns, lupus erythematosus, hyperpigmentation disorder, hypopigmentation disorder, skin cancer, skin infection, facial wrinkles, unattractive facial expressions, neck, multifunctional facial muscles, hypercontracted facial muscles, platysma girdles, and / or combinations thereof. (Item 54) Unwanted sweating, body odor, hyperhidrosis, bromhidrosis, chromatohidrosis, alopecia, Raynaud's phenomenon, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, lupus erythematosus, systemic lupus erythematosus, lupus discoid, drug-induced lupus, neonatal lupus, Crohn's disease, inflammatory bowel disease, ulcerative colitis, lung disease, asthma, chronic obstructive airway disorder, amyloidosis, systemic amyloidosis, cutaneous amyloidosis, cancer, skin cancer, hematological cancer, breast cancer, colon cancer, lung cancer, benign prostatic hyperplasia, dyslipidemia, hypercholesterolemia Methods for treating or preventing disorders selected from infectious diseases, C. difficile infection, Staphylococcus infection, dystonia, headache, pain, pain associated with arthritis, pain associated with rheumatoid arthritis, pain associated with psoriatic arthritis, pain associated with osteoarthritis, specified ophthalmic conditions, specified urinary tract conditions, neuromuscular disorders, conditions including muscle spasms and / or contractures, strabismus, hemifacial spasm, tremor, spasticity such as that resulting from multiple sclerosis, posterior orbital muscle, neurological conditions, Alzheimer's disease, Parkinson's disease, or stroke. (Item 55) The method according to any one of items 1 to 54, wherein the composition containing a large drug is formulated as a lotion, cream, powder, ointment, liniment, gel, or drop. (Item 56) A patch comprising an emulsion composition containing a large drug having a molecular weight of 100,000 Da or more, and a plurality of ultrafine needles. (Item 57) The patch according to item 56, wherein the composition containing the large drug compound comprises a nanoemulsion. (Item 58) The patch according to item 56, wherein the composition containing the large drug compound comprises a macroemulsion. (Item 59) The patch according to any one of items 56 to 58, wherein the needle has a length sufficient to protrude through the stratum corneum of the skin. (Item 60) The patch according to any one of items 57 to 59, wherein the needle is of insufficient length to reach the nerves in the dermis of the skin. (Item 61) The patch according to any one of items 57 to 60, wherein the needle has a length of approximately 10 to approximately 1000 μm. (Item 62) The patch described in any one of items 57 to 61, wherein the needle has a length of approximately 100 μm or more. (Item 63) The patch described in any one of items 57 to 62, wherein the needle has a length of approximately 300 μm or more. (Item 64) The aforementioned needle is made of a biocompatible material, and the patch is as described in any one of items 57 to 63. (Item 65) The aforementioned needle is made of metal, and the patch is as described in any one of items 57-64. (Item 66) The aforementioned needle is made of a soluble polymer, and the patch is as described in any one of items 57 to 64. (Item 68) The patch described in any one of items 57-66, wherein the aforementioned large drug is botulinum toxin. (Item 69) The patch described in any one of items 57-66, wherein the aforementioned large drug is an antibody drug. (Item 70) A kit comprising an emulsion composition containing a large drug having a molecular weight of 100,000 Da or more, and a patch containing multiple needles for use in applying the emulsion composition to a site. (Item 71) The kit according to item 70, wherein the emulsion composition is incorporated into the patch. (Item 72) A kit comprising an emulsion composition containing a large drug having a molecular weight of 100,000 Da or more, and a device for adjusting the microneedle at the site. (Item 73) The kit according to any one of items 70 to 72, wherein the emulsion composition comprises a nanoemulsion. (Item 74) The kit according to any one of items 70 to 72, wherein the emulsion composition comprises a macroemulsion. (Item 75) The device is a kit according to any one of items 72 to 74, comprising a plurality of needles. (Item 76) The kit described in any one of items 72-75, wherein the device is a patch, roller, stamp, or pen. (Item 77) A kit as described in either item 70 or 71, including the patch described in any one of items 56-69. (Item 78) The kit according to any one of items 70 or 72-77, wherein the emulsion composition is formulated as a lotion, cream, powder, ointment, liniment, gel, or drop.

Claims

1. A nanoemulsion composition for use in a method comprising applying the nanoemulsion composition to a skin site in combination with microneedle skin preparation (MSC) of the skin site, wherein the MSC is performed prior to administration of the nanoemulsion composition, wherein wherein the nanoemulsion comprises an oil dispersed within an aqueous dispersion medium to form oil particles, wherein a majority of the oil particles have a diameter of from about 10 nm to about 300 nm; and Where: (i) the nanoemulsion comprises a large drug having a molecular weight of 100,000 Da or greater, the large drug being or comprising a botulinum toxin; (ii) the nanoemulsion comprises an oil, and (iii) the nanoemulsion comprises a surfactant; wherein said oil and said surfactant are present in a ratio of 0.5 to 2.

0.

2. The composition for use according to claim 1 , wherein the botulinum toxin is delivered together with a biologically active agent.

3. A composition for use as described in claim 2, wherein the biologically active agent is selected from steroids, retinoids, anesthetics, fillers, silicones, collagen, hydrocortisone, retin A, and / or lidocaine.

4. The composition for use according to any one of claims 1 to 3, wherein the MSCs at the skin site are achieved with a device comprising a plurality of microneedles.

5. The composition for use according to claim 4, wherein the device is a batch, a roller, a stamp, or a pen.

6. 6. The composition for use according to any one of claims 1 to 5, wherein the skin site is a skin surface over a muscle or muscle group of a subject, a skin surface containing sweat glands, a skin surface containing sebaceous glands and / or a skin surface containing hair follicles.

7. 7. The composition for use according to any one of claims 4 to 6, wherein the microneedles have a length sufficient to protrude through the stratum corneum of the skin at the skin site and / or have a length insufficient to reach nerves in the dermis of the skin at the skin site.

8. The microneedle, (i) a length of about 10 to about 4000 μm; and / or (ii) a length of about 25 μm, about 100 μm, about 300 μm, about 500 μm, about 800 μm, about 1000 μm, about 1500 μm, about 2000 μm, or about 4000 μm or more; The composition for use according to any one of claims 4 to 7, comprising:

9. The composition for use according to any one of claims 4 to 8, wherein said microneedles are made of a biocompatible material, a metal or a dissolving polymer.

10. 10. The composition for use according to any one of claims 1 to 9, wherein the MSC comprises administration of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 microneedle (MN) or MN array impressions.

11. The composition for use according to claim 10, wherein said MN array is in the form of a stamp or roller and said impression is made by stamping or rolling.

12. The composition for use according to claim 10 or 11, wherein the MN array is rotated between one or more impressions.

13. 12. The composition for use according to claim 10 or 11, wherein the MN array is not rotated between one or more impressions.

14. A composition for use according to any one of claims 10 to 13, wherein said impressions are made on approximately the same sites, on overlapping sites or on different sites.

15. The blockbuster drug is (i) within about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 minutes of administration; (ii) within about 5 to about 60 minutes, about 5 to about 12 minutes, about 5 to about 15 minutes, or about 15 to about 30 minutes of administration; or (iii) within about 1, 2, 3, 4, 5, or 6 hours of administration. penetrates the skin to A composition for use according to any one of claims 1 to 14.

16. 16. The composition for use according to any one of claims 1 to 15, wherein the method comprises administering the emulsion composition in combination with MSCs at a lower dose compared to a reference topical treatment regimen without MSCs to produce a comparable therapeutic effect.

17. 17. The composition for use according to claim 16, wherein the reference topical treatment regimen comprises administering the emulsion composition without MSCs.

18. The composition for use according to any one of claims 1 to 17, wherein the method comprises administering multiple doses of the emulsion composition over time.

19. 20. The composition for use of claim 18, wherein said administering comprises administering a lower dose of said emulsion composition in combination with MSCs over a treatment period compared to a reference topical treatment regimen in which MSCs are not used to produce a comparable therapeutic effect.

20. 20. The composition for use of claim 19, wherein the reference topical treatment regimen comprises administering the emulsion composition without MSCs.

21. The composition for use according to any one of claims 18 to 20, wherein each dose of said emulsion composition is separated by a specific period of time.

22. 22. The composition for use according to claim 21, wherein the specified period of time is longer than the specified period of time for administering a reference local treatment regimen in which MSCs are not used.

23. The method may be used to treat unwanted sweating, body odor, hyperhidrosis, bromhidrosis, chromhidrosis, alopecia, Raynaud's phenomenon, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, lupus erythematosus, systemic lupus erythematosus, discoid lupus, drug-induced lupus, neonatal lupus, Crohn's disease, inflammatory bowel disease, ulcerative colitis, pulmonary disorders, asthma, chronic obstructive airway disorders, amyloidosis, systemic amyloidosis, cutaneous amyloidosis, cancer, skin cancer, blood cancer, breast cancer, colon cancer, lung cancer, prostatic hyperplasia, dyslipidemia, hypercholesterolemia, infectious diseases, C. or a method of treating or preventing a disorder selected from: C. difficile infection, Staphylococcus infection, dystonia, headache, pain, pain associated with arthritis, pain associated with rheumatoid arthritis, pain associated with psoriatic arthritis, pain associated with osteoarthritis, certain ophthalmological conditions, certain urological conditions, neuromuscular disorders, conditions involving muscle spasms and / or contractures, strabismus, hemifacial spasm, tremor, spasticity such as that resulting from multiple sclerosis, retroorbital muscles, neurological conditions, Alzheimer's disease, Parkinson's disease, or stroke; or the method is for treating a skin disorder or condition, the skin disorder or condition being selected from acne, unwanted sweating, body odor, hyperhidrosis, bromhidrosis, chromhidrosis, rosacea, alopecia, psoriasis, actinic keratosis, eczematoid dermatitis, disorders of excess sebum production, burns, lupus erythematosus, disorders of hyperpigmentation, disorders of hypopigmentation, skin cancer, skin infections, facial wrinkles, facial disfigurement, neck muscles, polyfunctional facial muscles, hypercontractile facial muscles, platysma, and / or combinations thereof; A composition for use according to any one of claims 1 to 22.

24. 1. A kit for delivering a large drug having a molecular weight of 100,000 Da or greater to a skin site, the kit comprising: (a) a nanoemulsion composition, wherein the nanoemulsion comprises an oil dispersed within an aqueous dispersion medium to form oil particles, wherein a majority of the oil particles have a diameter of from about 10 nm to about 300 nm; and Where: (i) the nanoemulsion comprises a large drug having a molecular weight of 100,000 Da or greater, the large drug being or comprising a botulinum toxin; (ii) the nanoemulsion comprises an oil, and (iii) the nanoemulsion comprises a surfactant; A nanoemulsion composition, wherein the oil and the surfactant are present in a ratio of 0.5 to 2.0; and (b) a device for microneedle preparation of the skin site, the device being used to prepare the skin site prior to administration of the composition; Including the kit.

25. 25. The kit of claim 24, wherein the device is a patch, roller, stamp, or pen.

26. 26. The kit of claim 25, wherein the device comprises a plurality of microneedles.

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