MX2008001230A

MX2008001230A - Microarray device

Info

Publication number: MX2008001230A
Application number: MXMX/A/2008/001230A
Authority: MX
Inventors: Nicholas Binks Peter; Marie Critchley Michelle; Alexander Irving Robert; William Pouton Colin; James White Paul
Original assignee: Nicholas Binks Peter; Marie Critchley Michelle; Alexander Irving Robert; Nanotechnology Victoria Pty Ltd; William Pouton Colin; James White Paul
Priority date: 2005-07-25
Filing date: 2008-01-25
Publication date: 2008-09-26

Abstract

A device is provided which is suitable for delivering at least one nanoparticle(s) to a subject. The device can be used to deliver a variety of nanoparticles, for example, therapeutic agents, directly through the outer layers of the skin without passing completely through the epidermis of the subject. Thus the device can be used to deliver therapeutic agents to a predetermined depth and avoid disturbing the pain receptors in the skin. Thus the device can be used to deliver agents, including therapeutic agents, in a non-invasive manner. A method of fabricating devices with associated nanoparticles is also provided.

Description

MICRO-ARRANGEMENT DEVICE FIELD OF THE INVENTION The present invention relates to a method and device for the delivery of nanoparticles. In particular, the present invention relates to microneedles and microneedle arrays suitable for the delivery of nanoparticles.

BACKGROUND OF THE INVENTION There has been increased interest in methods for the effective delivery of agents to organisms, including the delivery of therapeutic agents such as drugs. The delivery of agents to organisms is complicated by the inability of many molecules to penetrate biological barriers. Biological barriers for which it is preferable to deliver molecules through even the skin (or parts of it); the matoencephalic barrier h; mucous tissue (eg, oral, nasal, ocular, vaginal, urethral, gastrointestinal, respiratory); blood vessels; lymphatic vessels; or cell membranes (e.g., for the introduction of material into a cell or cells). Traditional methods of delivery such as oral administration are not suitable for all types of drugs because many drugs are destroyed in the digestive tract or immediately absorbed by the liver. Intravenous administration via hypodermic needles is also considered highly invasive and results in potential undesirable peak concentrations of the delivered drug. In addition, traditional delivery methods are often not useful for effective targeting of the delivered drug. One approach to the delivery of drugs through the skin is through the use of transdermal patches. A transdermal patch can provide significantly more effective blood levels of a beneficial drug because the drug is not delivered at peak concentrations as is the case with hypodermic injection and most oral administrations. In addition, drugs that are administered via transdermal patches are not subject to the severe environment of the digestive tract. Currently, transdermal patches are available for a specific number of drugs. Commercially available examples of transdermal patches include scopolamine for the prevention of motion sickness, nicotine as an aid to smoking cessation, nitroglycerin for the treatment of coronary angina pain, and estrogen for hormone replacement. Generally, these systems have drug pools sandwiched between an impermeable support and a type of membrane that constantly control the rate of delivery of the drug. Such patches depend on the ability of the drug to diffuse through the outermost layer of the skin, the stratum corneum, and eventually within the circulatory system of the patient. The stratum corneum is a complex structure of keratinized compact cell remnants that has an approximate thickness of 10-30 μm and forms an effective barrier to prevent the passage in and out of most substances. The degree of diffusion through the stratum corneum depends on the porosity of the skin, the size and polarity of the drug molecules, and the concentration gradient across the stratum corneum. These factors generally limit this mode of delivery to a very small number of useful drugs with very small molecules or unique electrical characteristics. A common method to increase the porosity of the skin is the formation of micropores or cuts through the stratum corneum. Many drugs can be administered effectively by penetrating the stratum corneum and delivering the drug to the skin at or below the stratum corneum. Devices for penetrating the stratum corneum generally include a plurality of microtable needles or sheets having a length to penetrate the stratum corneum without passing completely through the epidermis.

Examples of these devices are described in Pat. E.U. No. 5,879,326 to Godshall et al. , Pat. E.U. No. 5,250,023 to Lee et al and Pat. E.U. No. 6,334,856. Nevertheless, the efficacy of these methods to improve transdermal delivery has been limited, because after the micropores have been formed, it is necessary to administer the drug separately in the treated skin. In addition, these devices are usually manufactured with silicon or other metals using etching methods. For example, Pat. E.U. No. 6,312,612 to Sherman et al. describes a method for forming a microneedle array using Microelectromechanical Systems (MEMS) technology and standard microfabrication techniques. Although partially effective, the resulting microneedle devices are relatively expensive to manufacture and difficult to produce in large quantities. In addition, these arrangements have limited application in the delivery of a very limited variety of molecules.

SUMMARY OF THE INVENTION According to one aspect, the present invention provides a device suitable for delivering at least one nanoparticle comprising a microneedle having at least one nanoparticle that is associated with at least part of a surface of the microneedle and / or at least part of the tissue of the microneedle. The size of nanoparticle (s) may be in the range of about 1 to about 1000 nm. Preferably, the size of the nanoparticle may be between about 50 to about 500 nm. Preferably the device has at least two microneedles. The microneedles can be arranged in an order without pattern or other similar configuration. In other instrumentations, the microneedles can be arranged in at least one array. Preferably the nanoparticle (s) can be associated with at least part of the outer surface of the microneedle. Preferably the nanoparticle (s) can be associated with pores on the surface of the microneedles. In some instrumentations, the nanoparticle (s) may be associated with at least a portion of the tissue of the microneedle. The pore (s), cavities or the like can be of two or more shapes, cross sections that are selected from the group comprising circular, elongate, square, triangular, etc. In other instrumentations, the nanoparticle (s) can be associated with internal pores in the tissue of the microneedle. Preferably the association may comprise a covalent bond or non-covalent interactions. The non-covalent interactions can be selected from one or more of the group comprising ionic bonds, hydrophobic interactions, hydrogen bonds, Van der Waals forces or dipole-dipole bonds. Preferably the association is via a covalent link to a functional group in the microneedle. Preferably the functional group (s) can be selected from the group comprising COOR, CONR2, NH2, SH, and OH, where R comprises an H; organic or inorganic chain. The microneedle (s) can be manufactured from a porous or non-porous material that is selected from the group comprising metals, natural or synthetic polymers, glasses, ceramics, or combinations of two or more thereof. With this instrumentation, the polymer can be selected from the group comprising: polyglycolic acid / polylactic acid, polycaprolactone, polyhydroxybutyrate-valerate, polyorthoester, and polyethylene oxide / polybutylene terephthalate, polyurethane, silicone polymers, and polyethylene terephthalate, polyamine plus a sulphate tri-layer of dextran, high molecular weight poly-L-lactic acid, fibrin, methyl methacrylate (MMA) (hydrophobic, 70 mol%) and 2-hydroxyethyl methacrylate (HEMA) (hydrophilic 30 mol%), poly elastomeric polymers amide) (co-PEA), polyetheretherketone (Peek-Optima), biocompatible thermoplastic polymer, conductive polymers, polystyrene or combinations of two or more thereof. The microneedles may include a layer or coating on at least a portion of the surface of the microneedle (s) of an electrically conductive material. Preferably the electrically conductive material can be selected from the group comprising conductive polymers; Compound conductive materials; doped polymers, conductive metallic materials or combinations of two or more thereof. The conductive polymer can be selected from the group comprising substitutable or irreplaceable polymers comprising polyaniline; polypyrrole; polysilicones; poly (3, 4-ethylenedioxythiophene); polymer doped with carbon nanotubes; polymer doped with metal nanoparticles, or combinations of two or more thereof. Preferably the thickness of the layer or coating can be from about 20 nm to about 20 μm. The electrically conductive material can be layered or coated on the microneedle (s) by electrodeposition. At least one nanoparticle can be contained in the electrically conductive material. Preferably the nanoparticle (s) can be delivered to an organism and the microneedle (s) can be manufactured from a biocompatible material, the microneedle (s) can (even) be non-biodegradable (s). The microneedle can be solid. The microneedle may have nanosize pores or cavities on its surface. The nanoparticle (s) can (are) an active agent (s). In other instrumentation, the nanoparticle (s) can be a carrier for an agent. Preferably the nanoparticle may be associated with an active agent. The active agent (s) can be associated with the nanoparticle (s) by a covalent bond or non-covalent interactions. The non-covalent interactions can be selected from any or any of the group comprising ionic bonds, hydrophobic interactions, hydrogen bonds, Van der aals forces or dipole-dipole bonds. The nanoparticle can encapsulate the active agent. In another instrumentation, the active agent can be incorporated into the nanoparticle (s). Preferably the nanoparticle (s) can be made from a material that is selected from the group comprising metals, semiconductors, inorganic or organic polymers, magnetic colloidal materials, or combinations of two or more thereof. The metal can be selected from the group comprising gold, silver, nickel, copper, titanium, platinum, palladium and its oxides or combinations of two or more thereof. The polymer can be selected from the group comprising a conductive polymer; a hydrogel; agarose; polyglycolic acid / polylactic acid; polycaprolactone; polyhydroxybutyrate-valerate; poliortoester; polyethylene oxide / polybutylene terephthalate; polyurethane; polymeric silicon compounds; polyethylene terephthalate; polyamine plus a trilayer of dextran sulfate; high molecular weight poly-L-lactic acid; fibrin; copolymers of methyl methacrylate (MMA) and 2-hydroxyethyl methacrylate (HEMA), poly (ester-amide) elastomeric polymers (co-PEA); n-butyl cyanoacrylate; polyetheretherketone (Peek-Optima); polystyrene or combinations of two or more thereof. Preferably the active agent can be a biological agent. With this instrumentation, the biological agent can be a therapeutic and / or diagnostic agent. Preferably the therapeutic agent can be selected from the group comprising all microorganisms, viruses, viruses such as particles, peptides, proteins, carbohydrates, nucleic acid molecules, an oligonucleotide fragment (s) or DNA or RNA, lipids, organic molecules, molecules biologically active inorganics or combinations of two or more thereof. Preferably the therapeutic agent can be a vaccine. The vaccine can be selected from the group comprising a vector containing a nucleic acid, oligonucleotide, expression gene as a vaccine or combinations of two or more thereof. Preferably the vaccine can be selected from proteins or peptides as vaccines for diseases that are selected from the group comprising Johnes disease, hepatic dysphagia, bovine mastitis, meningococcal disease.

The vaccine may comprise Johnes peptide disease. With this instrumentation, the peptide can be selected from the group comprising: NVESQPGGQPNT (SEQ ID No 1); QYTDHHSSLLGP (SEC ID No 2); LYRPSDSSLAGP (SEC ID No 3); and / or its variants. The vaccine may comprise peptides from bovine mastitis disease. With this implementation, the peptide may be selected from the group comprising: MKKWFLILMLLGIFGCATQPSKVAAITGYDSDYYARYIDPDENKITFAINVDGF VEGSNQEILIRGIHHVLTDQNQKIVTKAELLDAIRHQMVLLQLDYSYELVDFAP DAQLLTQDRRLLFANQNFEESVSLEDTIQEYLLKGHVILRKRVEEPITHPTETAN IEYKVQFATKDGEFHPLPIFVDYGEKHIGEKLTSDEFRKIAEEKLLQLYPDYMID QKEYTIIKHNSLGQLPRYYSYQDHFSYEIQDRQRIMAKDPKSGKELGETQSIDN VFEKYLITKKSYKP (SEQ ID No 4); ILIRGIHHVL (SEC ID No 5); IRHQMVLLQL (SEC ID No 6); and / or its variants. The vaccine may comprise a peptide of Meningococcal disease. With this instrumentation, the peptide can be selected from the group comprising: GRGPYVQADLAYAYEHITHDYP (SEQ ID NO 7) STVSDYFRNIRTHSIHPRVSVGYDFGGWRIAADYARYRK NDNKYSV (SEQ ID NO 8); and / or its variants. The vaccine may include a Hepa ti tiis Virus C. With this instrumentation, the peptide can be selected from the group comprising: QDVKFPGGGVYLLPRRGPRL (SEQ ID No. 9); RRGPRLGVRATRKTSERSQPRGRRQ (SEC ID No 10); PGYPWPLYGNEGCGWAG LLSPRGS (SEC ID No 11); and / or its variants. The diagnostic agent can be a detectable agent. Preferably the detectable agent is used in an analysis. The outer diameter of the microneedle (s) may be between about 1 μm and about 100 μm. The length of the microneedle (s) can be between about 20 μm and 1 mm. Preferably the length of the microneedle (s) can be between about 20 μm and 250 μm. Preferably the microneedle (s) can be adapted to provide an insertion depth of at least about 100 to 150 μm. Preferably the shape of the tip of the microneedle (s) can be selected from the group comprising the square, circular, oval, cross needle, triangular, chevron, serrated chevron, half moon or diamond shapes. In an instrumentation, the complete microneedle can be manufactured from nanoparticles. According to another aspect, the present invention provides a method for manufacturing a device for the delivery of nanoparticles, the device comprises an array of microneedles and at least one nanoparticle that is associated with at least part of a surface of the microneedle, the method comprising the steps of: (i) covering at least a portion of the surface of a microneedle array that is molded with the nanoparticles; (ii) molding the microneedles; where after leaving the mold, the nanoparticles are associated with the surface of the microneedles. In still another aspect, the present invention provides a method for manufacturing a device for delivery of nanoparticles, the device comprising an array of microneedles and at least one nanoparticle that associates with the pores on the surface of the microneedle, the method comprising the steps of: i) inducing porosity on at least a portion of the surface of the microneedles; ii) associate the nanoparticles with at least a part of the pores. Preferably the step of inducing a porosity on the surface of the microneedles comprises the steps of: i) selectively leaching micro or nanoparticles that are incorporated into the surface of the microneedle; ii) give physical, chemical or electrochemical treatment to the surface of the microneedles. In a further aspect, the present invention provides a method for manufacturing a device for the delivery of nanoparticles, the device comprising an array of microneedles and at least one nanoparticle that is associated with at least part of the tissue of the microneedle, the method comprising the steps of: molding the microneedles in the presence of the nanoparticles; wherein after leaving the mold, the nanoparticles are associated with at least part of the tissue of the microneedles. In another additional aspect, the present invention provides a method for manufacturing a device for the delivery of nanoparticles, the device comprising an array of microneedles and at least one nanoparticle that is associated with at least a portion of the outer surface of the microneedle, the method comprising the steps of: (i) functionalizing at least a portion of the outer surface of the microneedles with functional groups; (ii) linking the nanoparticles to the introduced functional groups. Preferably the step of functionalizing can be selected from the group comprising oxidation, reduction, substitution, crosslinking, plasma, heat treatment or combinations of two or more thereof. Preferably the functional group (s) introduced (s) can be selected from the group comprising COOR, CONR2, NH2, SH, and OH, where R comprises a H or an organic or inorganic chain. The methods of the present invention may include the step of coating at least a portion of the microneedles with an electrically conductive material. Preferably the electrically conductive material can be selected from the group comprising the conductive polymer; composite conductive material; doped polymer, metallic conductive materials or compounds thereof. Preferably the conductive polymer can be selected from the group of substitutable or irreplaceable polymers comprising polyaniline; polypyrrole; polysilicone; poly (3, -ethylenedioxythiophene); polymer doped with metal nanoparticles; or polymer doped with carbon nanotubes.

In a further aspect, the present invention provides a device suitable for delivering at least one agent comprising a microneedle that is made of an electrically conductive polymer and / or electrically conductive polymer composite, the microneedle having at least one agent which is associated with at least part of a surface of the microneedle and / or at least part of the tissue of the microneedle. In a further aspect, the present invention provides a device suitable for delivering at least one agent comprising a microneedle that is made of an electrically conductive material, the microneedle having at least one agent that is associated with at least part of a surface of the microneedle and / or at least part of the tissue of the microneedle. The present invention also provides methods for using the microneedles to deliver nanoparticles. Thus according to another aspect, the present invention provides a method for delivering at least one nanoparticle (s) to a patient, wherein the delivery includes the steps of contacting at least one area of the patient with at least one microneedle associated with at least one nanoparticle, wherein at least one nanoparticle is delivered to the patient.

BRIEF DESCRIPTION OF THE FIGURES Figure 1 shows a schematic view of the cross sections of the needle. Figure 2 shows a top view of PDMS microneedles with dye molecules that are added to dye the patches and microneedle. Figure 3 shows a side view of the crosses shown in Figure 2. Figure 4 shows a side view of a microneedle array, the needles have a diameter of 20μm at the base and a degree of inclination of 50μm. Figure 5 shows a top view of a sheet of multiple microneedle array patches. Figure 6 shows an enlarged side view of a section of the array patch shown in Figure 5. Figure 7 shows a schematic flow diagram of a process for forming nanopore (s) on the surface of a microneedle. Figure 8 shows a fluorescent image of an array of circular microneedles showing the coverage of the quantum dot coating. Figure 9 shows a fluorescent image of a cross-shaped array of microneedles showing the coverage of the quantum dot coating. Figure 10 shows a scanning electron micrograph (SEM) image of insulin nanoparticles in PLGA microneedles. Figure 11 shows an SEM image of a microneedle array that is coated with insulin nanoparticles. Figure 12 shows a fluorescent confocal microscopy image of a skin patch that is removed from a hairless mouse. Figure 13 shows a fluorescent image of confocal microscopy at a total depth of approximately 60 μm.

DETAILED DESCRIPTION OF THE INVENTION The devices described herein are useful for transporting agents to or through biological barriers that include the skin (or parts thereof); the blood-brain barrier; mucosal tissue (eg, oral, nasal, ocular, vaginal, urethral, gastrointestinal, respiratory); blood vessels; lymphatic vessels; or cell membranes (e.g., for the introduction of material into a cell or cells). Biological barriers can be found in humans or other types of animals, as well as in plants, insects, or other organisms, which include bacteria, yeast, fungi, and embryos. The microneedle devices can be used in internal tissues with the help of a catheter or laparoscope. For certain uses, such as for delivery of drug to an internal tissue, the device can be surgically implanted. The present invention provides agents that can be a protein, peptide, homogenized cell, whole organism or glycoprotein effective as a detecting agent or protective agent. The present invention also provides a presentation configuration of the agent that can be used to detect, individual molecules, multimer, aggregates, or multimer through the nanoparticle anchor; in consideration of that, to deliver (vaccination) The configuration of the biological molecule can also comprise: individual molecules, multimer, aggregates, or multimer through the nanoparticle anchor. The nanoparticle anchor can be through nanoparticles of gold, silver, titanium, agarose, proteins, dendrimers, proteins or polymers. The preferred option is the presentation of multimeric nanoparticle.

The present invention also has applications in the food industry for quality detection and for one or more infectious agent (s), the infectious agent can be a microorganism. The microorganism can be selected from one or more of the group comprising viruses, bacteria, protozoa and / or fungi. The inventors of the present invention have unexpectedly discovered a novel delivery structure and composition, as well as the composition and configuration of the biological reagent to be delivered and methods for its production. By forming the agents to deliver in the presence of removable and / or degradable nanoparticles of different composition to the composition of the delivery molecules, the nanostructured molecules incorporate a nanoporous structure capable of retaining large and small molecules and nanoparticles - biological molecules that are anchored to be delivered as vaccines and therapeutic. It will also be recognized that a novel polymer system number that when subjected to a certain tension changes its composition to have a nanoparticular structure that is different from the surrounding polymer, and said polymers can have an application with its improved solubility (degradation properties). for the delivery of reagents from polymer array patches. The mentioned polyvalent nanoparticular vaccination particles can be released from polymer patches with the penetration of the interstitial layer into living tissue. The polyvalent nanoparticular detector agents mentioned can be retained on the surface of the polymer patches with conductive properties for signal transduction. The authors of the present invention have surprisingly found that the identical polymer is used to present (delivery / anchor detector) the nanostructured molecule (s), and also unexpectedly, a polymer which, although is biocompatible, preferably it is not biodegradable and therefore has advantages of molecule delivery speed without requiring long-term dependent degradation. In the aspect of the present invention that has application for the delivery of vaccination through the stratum corneum, the residence time in this layer is of an estimated time of two weeks. In a further aspect of the present invention a process is provided for delivering molecule (s) precisely to the proper depth using the microneedle arrays that have nanostructured delivery molecules. The construction of the device and control of the polymer structure is carried out by means of the inclusion of materials with nanoparticle size with properties to allow the dissolution of the nanoparticles to create a mesoporous structure with nanoporous cavities to retain reagents or structured nanoparticle reagents , which are delivered by means of the structure of the fix patch. Both hollow and solid penetration arrays (solid needle) are constructed with any of a variety of sizes between 20 μm and 250 μm but the preferred sizes (lengths) are 25 μ and 150 μm. The dimensions of the complete arrangement could normally be 1 square cm or with a diameter of 1 cm. However, the size of the patch patch can be based on the amount of material being delivered and the needle density that is packed in the patches. It is preferable that the microneedles be in an array format, but they can be arranged randomly. The disposition of the microneedles can be a result of the method used in manufacturing. The microneedles can be arranged in such a way that more than one reagent can be coated and delivered from the array itself. A polymer that when subject to certain stresses changes its composition to have a nanoparticle structure that is different from the surrounding polymer, and said polymers can have an application with their improved solubility (degradation properties) for the delivery of reagents from polymer array patches. A polymer that contains a nanoparticle that can be selectively removed to produce nano-size pores or cavities on the surface of the microneedle. The microneedle array patches of the present invention also provide applications for the treatment and prevention of human diseases. Preventive vaccination of a wide variety of human disease states can be achieved, for example, the present microneedle arrays can be used to vaccinate against any or all of the disease states that are selected from the group comprising infectious diseases (which include but they are not limited to meningococcal disease and tuberculosis) and autoimmune diseases (which include but are not limited to multiple sclerosis and rheumatoid arthritis). As used in the present invention, the term "nanoparticle" is intended to include particles comprising sizes from about 1 nm to about 1000 nm. Preferably, the nanoparticles are in the range of about 50 nm to about 500 nm. As used in the present invention, the term "tissue" is intended to describe the material of which the particle is composed. As used in the present invention, the term "biocompatible" is intended to describe molecules that are not toxic to cells. The compounds are "biocompatible" if their addition to in vitro cells results in less than or equal to 20% cell death and does not induce inflammation or other similar adverse effects in vivo. As used in the present invention, "associating" or "associated" includes physical, chemical, and physiochemical adhesion. As used in the present invention, "biodegradable" includes those compounds that, when introduced into cells, are broken down by the cellular machinery into components that the cells can either reuse or dispose of without any significant toxic effect on the cells (that is, just under 20% of the cells die). The agent that can be delivered using the present invention includes any therapeutic substance that possesses suitable therapeutic characteristics. These agents may be selected from any or any of the group comprising: thrombin inhibitors, antithrombogenic agents, thrombolytic agents, fibrinolytic agents, vasospasm inhibitors, calcium channel blockers, vasodilators, antihypertensive agents, antimicrobial agents, antibiotics, receptor inhibitors, surface glycoprotein, antiplatelet agents, antimitotic agents, icrotubular inhibitors, antisecretory agents, actin inhibitors, remodeling inhibitors, antisense nucleotides, antimetabolites, antiproliferatives, anti-cancer chemotherapeutic agents, steroidal anti-inflammatory agents or nonsteroidal anti-inflammatory drugs, immunosuppressive agents, growth hormone antagonists , growth factors, dopamine agonists, therapeutic radioagents, peptides, proteins, enzymes, components of the extracellular matrix, ACE inhibitors, free radical collectors, chelating agents, antioxidants, antipolymerases, antiviral agents, photodynamic therapy agents, and gene therapy agents. In particular, the therapeutic substance can be selected from any or any of the group comprising alpha 1 antitrypsin, antiangiogenesis agents, antisense, butorphanol, calcitonin and the like, ceredase, COX-II inhibitors, dermatological agents, dihydroergotamine, agonists and antagonists of dopamine, enkephalins and other opioid peptides, epidermal growth factors, erythropoietin and the like, follicle-stimulating hormone, G-CSF, glucagon, GM-CSF, granisetron, growth hormone and the like (which includes growth hormone releasing hormone), growth hormone antasts, hirudin and the like of hirudin such as hirulog, IgE suppressors, imiquimod, insulin, insulinotropin and the like, insulin-like growth factors, interferons, interleukins, luteinizing hormone, luteinizing hormone releasing hormones and the like, heparins, low molecular weight heparins and other natural, modified, or synthetic glycoaminoglycans, M-CSF, metoclopramide, midazolam, monoclonal antibodies, pegylated antibodies, pegylated proteins or any proteins that are modified with hydrophilic or hydrophobic polymers or functional groups additional, fusion proteins, fragments of single chain antibodies or the same with any combination of adhered proteins, macromolecules, or additional functional groups thereof, narcotic analgesics, nicotine, non-steroidal anti-inflammatory agents, oligosaccharides, ondansetron, parathyroid hormone and the like , ant parathyroid hormone asts, prostaglandin antasts, prostaglandins, recombinant soluble receptors, scopolamine, serotonin asts and antasts, sildenafil, terbutaline, thrombolytics, tissue plasminogen activators, TNF-, and TNF- antasts, vaccines, with or without carriers / adjuvants, including prophylactic and therapeutic antigens (including but not limited to subunit protein, peptide and polysaccharide, polysaccharide conjugate, toxoids, gene-based vaccines, live attenuated, reassortant, inactive, whole cells, viral vectors and bacterial) in connection with, addiction, arthritis, cholera, ***e addiction, diphtheria, tetanus, HIB, Lyme disease, meningococcus, measles, mumps, rubella, chicken pox, yellow fever, respiratory syncytial virus, Japanese encephalitis transmitted by ticks , pneumococcus, streptococcus, typhoid, influenza, hepatitis, which includes hepatitis A, B, C and E, oti ti s media, rabies, polio, HFV, parainfluenza, rotavirus, Epstein Barr virus, CMV, chlamydia, hemophilia without classification, moraxela catarrhalis, human papilloma virus, tuberculosis that includes BCG, rrhea, asthma, atherosclerosis of malaria, E-coli, Alzheimer's disease, H. pylori, salmonella, diabetes, cancer, herpes simplex, human papilloma and other similar substances that include all of major therapeutics such as agents for the common cold, antiadictions, anti-allergies, antiemetics, anti-obesity, antiosteoporotic, anti-infective, analgesics, anesthetics, anorexics, antiarthritics, anti-asthmatic agents, anticonvulsants, antidepressants, antidiabetic agents, antihistamines, anti-inflammatory agents, anti-migraine preparations, antineetomy preparations, antinausea, antineoplastics, antiparkinson drugs, antipruritics, antipsychotics, antipyretics, anticholinergics, benzodiazepine antasts, vasodilators, what in gene These include coronary, peripheral and cerebral agents, bone stimulation, central nervous system stimulants, hormones, hypnotics, immunosuppressants, muscle relaxants, parasympatholytic agents, parasympathomimetics, prostaglandins, proteins, peptides, polypeptides and other macromolecules, psychostimulants, sedatives, and hypofunction. sexual and tranquilizers.

Johne's Disease Paratuberculosis (Johne's disease) is a chronic, progressive and enteric disease of ruminants caused by infection with Mycobacterium for tuberculosis. Symptoms of the disease in infected animals include weight loss, diarrhea, and decreased milk production in cows. The prevalence of Johne disease in cattle is estimated at 22-40% and the economic impact of this disease in the dairy industry was estimated at more than $ 200 million per year in 1996. In addition, the M. for tuberculosis has been implicated as a causative factor of Crohn's disease, a chronic inflammatory bowel disease in humans, which has served as an additional boost to control this disease in our national cattle industry. The treatment and prevention of Johne's disease has made it a high priority disease in the cattle industry. Membrane protein p34, SEQ ID No IA, elicits the predominant humoral response against M. for tuberculosis and within the published sequence the antigenic peptide epitopes have been identified, including but not limited to: NYESQPGGQPNT (SEQ ID No 1) QYTDHHSSLLGP (SEC ID No 2) LYRPSDSSLAGP (SEQ ID No. 3) See for example, Ostro ski, M et al. (2003) Scandinavian Journal of Immunology, 58, 511-521. Peptide regions in other potential antigens can also be used in the device which can include the antigens described in: Hydroperoxide Reductases C and D Bishop which are major antigens constitutively expressed by Mycobacterium um avi um subsp. of paratuberculosis. Olsen, et al. (2000) Infection and immunity, 68 (2), 801-808. Two pi 1 and p 20 proteins have been identified as potential antigens to be used in vaccination. Thus, properly nanostructured vaccines for Mycobacterium um infection for diseases such as Johnes disease can be made and delivered according to the method and device of the present invention.

Bovine mastitis Bovine mastitis is a serious problem, common in lactating animals of both dairy and bovine types. The treatment for this disease is practiced mainly in the dairy type animal where it is required to handle udders daily. Mechanical milking machines may have caused an increased incidence of mastitis; The true origins of the disease remain unknown. The bacterial organisms that are identified in affected glands are varied; however, the species of Streptococcus and Staphlococcus are the most commonly isolated. Purified proteins that act as antigens for bovine mastitis have also been described and incorporated by reference; Immunization of dairy cattle with recombinant Streptococcus uberis GapC or a CAMP chimeric antigen provides protection against heterologous bacterial challenge. Fontaine et al. (2002) Vaccine, 2278-2286. It is expected that the specific peptide epitopes of these proteins would be antigenic. The PauA protein has been successfully used to vaccinate cattle to prevent mastitis caused by S challenge infection. uberis (Leigh, J. A. 1999. "Streptococcus uberis: a permanent barrier to the control of bovine mastitis?" Vet. J. 157: 225-238). Vaccinated, protected cattle generated serum antibody responses that inhibited plasminogen activation by PauA. , PauA protein sequence S. uberis: MKKWFLILMLLGIFGCATQPSKVAAITGYDSDYYARYIDPDENKITFAINVDGFVEGSN QEILIRGIHHVLTDQNQKIVTKAELLDAIRHQMVLLQLDYSYELVDFAPDAQLLTQDRR LLFANQNFEESVSLEDTIQEYLLKGHVILRKRVEEPITHPTETAMEYKVQFATKDGEFH PLPIFVDYGEKHIGEKLTSDEFRKIAEEKLLQLYPDYMIDQKEYTIIKHNSLGQLPRYY S YQDHFSYEIQDRQRIMAKDPKSGKELGETQSIDNVFEKYLITKKSYKP (SEQ ID No 4) The peptides of the epitope region are selected from this protein are useful as vaccine candidates when presented in the form of appropriate nanoparticle: that includes but is not restricted to ILIRGIHHVL (SEC ID No 5) IRHQMVLLQL (SEC ID No 6) As well as the complete or selected fragments of the above-mentioned protein sequence.

Meningococcal disease The Omp85 proteins of the Neisseria gonorrhoeae and N. meningi tides and the peptide sequences that are derived therefrom can be used as vaccines against the organisms that cause meningococcal disease when they occur in nanoparticle form or variants, according to EU 2005074458, which is incorporated herein invention by reference. And the gonococcal and opacity proteins according to EP0273116, which include but are not restricted to: GRGPYVQADLAYAYEHITHDYP (SEQ ID No. 7) STVSDYFRNIRTHSIHPRVSVGYDFGG RIAADYARYRK NDNKYSV (SEQ ID NO 8) and its variants.

Hepatitis C Virus Fragments of the core protein used for in vitro immunization may include but are not limited to: QDVKFPGGGVYLLPRRGPRL (SEC ID No 9) RRGPRLGVRATRKTSERSQPRGRRQ (SEC ID No 10) PGYPWPLYGNEGCG AGWLLSPRGS (SEC ID No 11) These are can be used in conjunction with or without Toll and / or lipoprotein receptors as indicated in the following reference: Cell activation by synthetic lipopeptides of hepatitis C virus (HCV) - the core protein is mediated by similar receptors toll (TLRs) 2 and 4.

Hepatic Dysoma Liver fluke (Fasciola spp.) Infects a wide variety of animals, including humans. The disease that is caused is known as Fasciolosis. As with most parasitic diseases, it has a complex life cycle. Economically, sheep and cattle are of primary importance. Infection with liver fluke leads to decreased production due to poor energy conversion (meat and milk in cattle, meat and wool in sheep) and can lead to mortality (particularly in sheep). Targeted vaccines for the liver fluke have been investigated for many years, most subunit vaccines focusing on glutathione S-transferase (GST), cathepsin L (catL) and fatty acid binding proteins (FABP). Attenuated vaccines, which are created by metacercaria irradiation, are very effective, however this method of vaccination is not commercially viable. Therefore, candidates for subunit vaccines have been considered. DNA vaccines and recombinant proteins such as cathepsin B have been cloned and analyzed. Antigens and the use of cathepsin L proteases have been cloned as the vaccines described, see for example EU Patents No. 6,623,735 and 20050208063, which are incorporated herein by reference. The N-terminal sequences of the proteases to be used for in vitro immunization may include but are not limited to: AVPDKIDPRBSG (SEQ ID NO: 12) These may be incorporated into a nanoparticle (s) or may be formed as a nanoparticle .

Injectable nanoparticles An injectable nanoparticle can be prepared with that which includes a substance that is delivered and a nanoparticular polymer that covalently limits with the molecule (s), where the nanoparticle is prepared in such a way that the molecule delivery (s) ) is carried out on the outer surface of the particle. Nanostructured molecules injectable for example with antibodies or fragments of antibodies on their surfaces can be used to target specific cells or organs as preferred for the selective dosing of drugs. The molecule to be delivered can covalently limit the nanoparticular polymer by reaction with a terminal functional group, such as the hydroxyl group of a poly (alkylene glycol) nanoparticle by any method known to those skilled in the art. For example, the hydroxyl group can react with a terminal carboxyl group or amino terminal group on the molecule or antibodies or antibody fragment, to form an ester or amide bond, respectively. Alternatively, the molecule can be linked to poly (alkylene glycol) through a dysfunctional space group such as diamine or dicarboxylic acid, which include but are not limited to sebacic acid, adipic acid, isophthalic acid, terephthalic acid, fumaric acid, dodecanedicarboxylic acid, azelaic acid, pimelic acid, suberic acid (octanedioic acid), itaconic acid, biphenyl-4,4 '- dicarboxylic acid, benzophenone-4,4'-dicarboxylic acid, and p-carboxyphenoxyalkanoic acid. In this embodiment, the space group reacts with the hydroxyl group on the poly (alkylene glycol), and then reacts with the molecule (s). Alternatively, the space group may react with the molecule, such as an antibody or antibody fragment, and then react with the hydroxyl group on the poly (alkylene glycol). The reaction should be carried out under conditions that do not adversely affect the biological activity of the molecule that adheres covalently to the nanoparticle. For example, those conditions that cause the denaturation of proteins or peptides, such as high temperature, certain organic solvents and solutions of high ionic strength, should be avoided when a protein is bound to the particle. For example, organic solvents can be removed from the reaction system and a water-soluble coupling reagent such as EDC used instead. According to another embodiment, the agent that is delivered can be incorporated into the polymer acting as a nanoparticle formation. Substances to be incorporated should not chemically interact with the polymer during manufacture, or during the release process. Additives such as inorganic salts, BSA (bovine serum albumin), and inert organic compounds can be used to alter the release profile of the substance, as is known to those skilled in the art. Biologically labile materials, for example, prokaryotic or eukaryotic cells, such as bacteria, yeast, or mammalian cells, including human cells, or components thereof, such as cell walls, or cellular conjugates, may also be included in the particle. The injectable particles that are prepared according to this process can be used to deliver drugs such as non-steroidal anti-inflammatory compounds, anesthetics, chemotherapeutic agents, immunotoxins, immunosuppressive agents, steroids, antibiotics, antivirals, antifungals, and spheroidal anti-inflammatories, anticoagulants. For example, hydrophobic drugs such as lidocaine or tetracaine can be entrapped in the injectable particles and released for several hours. Charges in nanoparticles as high as 40% (by weight) can be achieved. Hydrophobic materials are more difficult to encapsulate, and in general, the loading efficiency decreases over that of a hydrophilic material. In one embodiment, an antigen is incorporated into the nanoparticle, alternatively, the antigen can make up the nanoparticle in its entirety. The term "antigen" includes any chemical structure that stimulates the formation of antibodies or elicits a cellular mediated humoral response, including but not limited to protein, polysaccharide, nucleoprotein, lipoprotein, synthetic polypeptide, or a small molecule (hapten) linked to a protein carrier. The antigen can be administered together with an adjuvant as preferred. Examples of suitable adjuvants include synthetic glycopeptides, muramyl dipeptide. Other adjuvants include Bordetella pertussis dead, the liposaccharide of Gram-negative bacteria, and large polymeric anions such as dextran sulfate. A polymer, such as a polyelectrolyte, can also be selected for the manufacture of the nanoparticle that provides adjuvant activity. Specific antigens that can be loaded into the nanoparticles described herein include, but are not limited to, attenuated or killed viruses, toxoids, polysaccharides, cell wall or surface or virus and bacteria coating proteins. These can also be used in combination with conjugates, coadjuvants, or other antigens. For example, Haemophilius influenzae in the form of purified capsular polysaccharide (Hib) can be used alone or as a conjugate with diphtheria toxoids. Examples of organisms from which these antigens are derived include poliovirus, rotavirus, hepatitis A3 B, and C, influenza, rabies, HIV, measles, mumps, rubella, Bordetella pertussus, Streptococcus pneumoniae, Clostridium diptheria, C. tetani, Vibrio cholera, Salmonella spp. , Neisseria spp. , and Shigella spp. . The nanoparticle must contain the substance that is delivered in an amount sufficient to deliver to a patient an amount of therapeutically effective compound, without causing serious toxic effects in the patient being treated. The desired concentration of active compound in the nanoparticle will depend on the rates of absorption, inactivation, and excretion of the drug as well as the rate of delivery of the compound from the nanoparticle. It will be noted that the dose values will also vary depending on the severity of the condition to be alleviated. It will further be understood that for any particular subject, the specific dose regimens should be adjusted over time according to the individual need and professional judgment of the person administering or supervising the administration of the compositions. Now, the present invention will be more fully described with reference to the appended examples. It will be understood, however, that the description below is for illustrative purposes only and should not be taken in any way as a restriction on the generalities of the present invention described above.

EXAMPLE 1 Formation of a mold using a polycarbonate sheet Laser ablation A polycarbonate sheet is ablated by laser using an excimer laser beam. The cross section of the needle is determined by the shape of the aperture through which the laser beam passes before the polycarbonate workpiece is irradiated. This process, known as excimer laser photolithographic ablation, uses an imaging lens to form the desired shapes. The depth of the laser ablation, and therefore the maximum height of the mold material, are determined by a computer program that operates the excimer micromachining system. Using the excimer laser ablation of a polycarbonate sheet, a series of molds for microneedle arrays with eleven different shapes and heights and of a variety between 20μm and 200μm were manufactured. The molds were manufactured for a number of different forms of microneedles including square, circular, oval, cross needle, triangular, chevron, serrated chevron and half moon. In addition to the shape of the micro-needle, the density, depth and degree of inclination of the microneedle were varied. For example, the laser ablation process was used to create molds for two dense arrays: a) forms with 50μm diameter with a degree of inclination of 50μm and approx. lOOμm in height. b) shapes with 100 μm diameter with a degree of inclination of 100 μm and approx. 100 μm in height. The molds were evaluated to determine their suitability for the manufacturing process with a variety of techniques including optical microscopy, laser scanning, confocal microscopy and electron scanning microscopy. In our experience, structures with good perforations are usually complex in cross sections, and not normal simple conical protrusions. Therefore, the shapes were chosen among those that contain characteristics and symmetry of edges such that they lead to improve the performance of perforations.

EXAMPLE 2 Fabrication of microneedle arrays The first molding tests were carried out with materials with two different viscosities. The more viscous material had a consistency similar to a putty, the second had a viscosity similar to honey. These materials were applied to the polycarbonate molds and pressure was applied by means of a glass tile to ensure that the indentations were filled. To assist in the removal of gas bubbles in the molds, vacuum was applied to the molding materials. The material was hardened by curing the polymer / polymer precursor using a sixty-second exposure to light from a portable source of blue LED through the glass tile. The extraction of the mold was a simple process, relying on the tendency of the material to a greater adhesion to the back of the glass tile more than to the polycarbonate mold. The molds were made with polycarbonate sheet with 250 to 500 μm thickness and were more flexible than glass tile. Therefore the molded material could be "peeled off" from the more slightly flexible mold. The resulting structures were examined under an optical microscope. Some of the structures were measured using a confocal laser scanning microscope or projected using a scanning electron microscope.

Results The second honey-like material filled the mold, and the air bubbles that formed in the needle in the nooks and crannies of the mold were eliminated through the application of vacuum. Many of the structures were successfully removed from the mold and the mold was recovered for future tests with a combination of liquid and sonication. A silicone releasing agent was applied to the polycarbonate to assist in the extraction of the mold, alternatively, materials such as PEEK or silicone elastomers can be used as concave molds.

EXAMPLE 3 Manufacture of various microneedle arrays A number of micro-needle arrays were manufactured with variation in shape, length, aspect ratios and needle densities. The various forms are shown in figure 1. i) Cross-shaped needle approximately 170μm in height Cross-shaped needle molds filled well with polymer, including the point at the intersection of the cross that is formed as a result of the ablation process. The combination of the relatively large lateral arms and the fine characteristic at the apex produces a robust structure with good mechanical properties. i) Circular microneedle with 50μm diameter The circular microneedle was produced approximately 140μm in height with an approximate aspect ratio of 3. ii) Triangular microneedle 50μm per side A triangular microneedle was prepared with approximately 100 μm height and an aspect ratio of approximately 2. The smooth apex of the shape is due to the polymer molding material and does not fully reproduce the fine texture of the ablation mold. iii) Circular microneedles Arrangement patches were produced with circular microneedles 20 μm in diameter and 50 μm in height and 100 μm in diameter with a degree of inclination of 100 μm, and approximately 100 μm in height. v) Oval shapes, cheiron, chevron toothed, triangle, crescent and diamond A variety of different needle-shaped profiles were produced to investigate the effect of perforation on the skin of the microneedle shape.

EXAMPLE 4 Fabrication of patch patches with spikes and color crossings Patch patches were constructed with a series of colored tips and crosses with polydimethylsiloxane (PDMS), a clear elastomer material by excimer laser machining 2 polycarbonate molds with four patches of 10 mm x 10 mm each, with concave features of circular reduction structures, and crosses. The degree of inclination and depth of the structures were varied. Clear and color PDMS were projected from these characteristics. The first modeling tests were conducted with standard PDMS supplied by DUPONT. This is a two-part formulation, adding 10% accelerator to cause the material to set. The mixture was placed in a vacuum chamber to accelerate degassing before molding to prevent bubble formation during curing. Figure 2 shows a top view of the manufactured cross-shaped microneedles PDMS and Figure 3 shows the side view of the manufactured cross-shaped microneedles. Figures 4, 5 and 6 show various microneedle arrays prepared according to the methods described. An aqueous based dye was added to the PDMS before molding; the addition of large amounts of dye intensifies the color, additional hardening accelerator was added to compensate for the volume of added aqueous dye. The material was hardened by curing the molded material by placing it in an oven at 45 ° C for several hours. The cure rate was significantly slower for the material with color. Surprisingly upon leaving the mold the water-colored material was more successful than the colorless material. This may be due to a variety of effects such as that of the hardening accelerator, molding with thicker parts that tend to be held within the needles more effectively during the extraction of the mold, or perhaps some inhibition of adhesion between the PDMS and the polycarbonate as a result of the aqueous additive.

EXAMPLE 5 Post-cure modification of the microneedle arrays The microneedles that were produced with the method of Example 3 can be coated with a layer of a biocompatible electrically conductive polymer to modify delivery characteristics of the microneedle. In this way, to assist in the delivery of certain types of molecules, a polyaniline coating can be applied to the solid polymeric microneedle after leaving the mold. The conductive polymer can be employed using techniques known in the art, including electrodeposition. During the electrodeposition phase (which includes polymerization) the biological reagents (for vaccines, drug delivery, etc.) can be included in the conductive polymer. The conductive polymer can be polymerized (electrodeposited) under conditions such that the surface of the electrodeposited polymer has characteristics that allow diffusion of the biological reagent to the outside in the surrounding environment (skin) so that the biological reagent is functional for its purpose. A different number of coatings can be produced. Thicknesses can be employed depending on the desired application, ranging from 20 nm to 20 μm. In another experiment, polyaniline and polypyrrole can be co-deposited electrochemically in microneedles made of conductive materials under potentiostatic or galvanostatic conditions. The electropolymerization can be carried out by varying the applied potential and the feed rate of the monomers. The formation of polyaniline-polypyrrole compound coatings can be confirmed by the presence of characteristic peaks in polyaniline and polypyrrole in the infrared spectrum. The coatings composed of polyaniline and polypyrrole can be formed in the applied potentials of < 1.0 V. Polypyrrole is preferably formed at 1.5 V. Electrodeposition methods have been previously described and include Adeloju, S.B. and Shaw, S. J., (1993) "Potentiometric biosensor with polypyrrole base for urea" Analytica Cimica Actica, 281, pages 611-620; Adeloju S.B. and La al, A., (2005) Intern. J. Anal. Chem. , 85, pages 771-780, based on its use as a sensor. We have surprisingly discovered that the techniques can be used for the incorporation of proteins and peptides in a polymer layer to deliver proteins and peptides as a therapeutic as well as peptide and protein antigens (for vaccines), hormones (erythropoietin, parathyroid hormone) and drugs (insulin) EXAMPLE 6 Delivery Nanoparticles Nanoparticles can be formed with metals (gold silver) light metals, polymer material by any of the standard techniques (U.S. Patent No. 6,908,496 Halas et al; U.S. Pat. No. 6, 906, 339 Dutta; U.S. Pat. No. 6,855,426 Yadav; U.S. Pat. No. 6,893,493 to Cho et al.). The surface of the nanoparticles can be functionalized to anchor / immobilize (multimerize) the biological reagents to improve the efficacy of the immunization. Other examples without limitations of methods for the formation of nanoparticles include: Cao L, Zhu T and Liu Z (2005) "The formation of a mechanism of non-spherical gold particles during inoculation growth: function of adsorption of anion and speed of reduction . " Colloid Interface Science Journal, July 11. Bilati U, Alleman E and Doelker E. (2005) "Poly (D, L-lactide-co-glycolide) nanoparticles with protein loading prepared by means of the double emulsion method - Processing and formulation issues to increase adhesion efficiency. " Microencapsulation Journal, 22 (2), 205-214. Rolland JP, Maynor BW, Euliss LE, Exner AE, Denison GM and Desimone JM (2005) "Direct manufacturing and collection of monodisperse nanobiomaterials, specifically." Journal of the American Chemical Society, 127 (28), 10096-100. Biological agents can be immobilized on the surface of a nanoparticle or integrally incorporated into the nanoparticle during manufacture. The delivery agent can even be directly manufactured or naturally presented in a nanoparticular form. The biological agents insulin and ovalbumin were structured as nanoparticles using supercritical fluid technology, to produce nanoparticles of dimensions 50-300 nm. The insulin nanoparticles were suspended in a solvent (ethanol) and adhered to the surface of the microneedles. The insulin and ovalbumin that adhered to the microneedles are each delivered separately through the stratum corneum and the response to insulin delivery can be measured. Erythropoietin is a glycoprotein hormone that is produced in the liver during fetal life and in the kidneys of adults and is involved in the maturation of erythroid progenitor cells in erythrocytes. There are several human conditions and treatments for cancer which results in low levels of circulating red blood cells and therefore the administration of erythropoietin is preferred. The erythropoietin can be nanostructured by means of supercritical fluid technology and added to the microneedles to be delivered by means of a microneedle array, and the efficiency of delivery can be measured by the physiological effects on the number of red blood cells in mice (including flow cytometry). EXAMPLE 7 Nanoparticles for the creation of nanopores in array patch microneedles The surface of a polymeric microneedle array can be nanostructured during fabrication by means of a microneedle liner that is molded with nanoparticles that can be removed selectively. The microneedles can then be molded, hardened and extracted from the mold to produce microneedles with nanoparticles that adhere to the surface of the microneedles. The adhered nanoparticles can then be removed, for example by dissolution or extraction techniques, to produce a microneedle having nanosize pores or cavities on its surface. The molecules or nanoparticles of the delivery agent can then be associated with the introduced pores by means of non-covalent interactions or covalent bonds. With respect to the process shown in Figure 7, the method includes the steps of: (i) "Stenciling" the soluble nanoparticles that are incorporated into microneedles during the manufacture of patches; (ii) Place in the template the nanoparticles that are removed with solvent leaving recesses on the surface of the microneedle and then add nanostructured reagent (s) to the solution; (iii) The nanostructured reagent (s) fit within the recesses within the needle structure to form the microneedles with the nanostructured reagents that are associated with the microneedles. The molded microneedle can alternatively be chemically treated with a solvent, chemical reagent, electrochemical or physical treatment to induce the surface cavity and / or nanopore formation.

EXAMPLE 8 Microneedles made of electrically conductive polymers A polyaniline microneedle array can be manufactured by electropolymerizing a monomer solution contained in a microneedle array mold under an applied potential. The progress of electropolymerization can be monitored by weight gain analysis and infrared spectroscopy. The nanoparticles can be added to the monomer solution before polymerization to form a microneedle array with the delivery molecule integrally incorporated into the needles, or the nanoparticles can be associated with the surface of the microneedles by means of a subsequent step to the output of the mold.

EXAMPLE 9 Coating of quantum dots in microneedle arrays To demonstrate the efficiency of nanoparticle patch loads, a series of microneedle arrays were coated with quantum dots. The quantum dots are semiconductor crystals typically between 1 and 10 nm in diameter and have unique properties between those simple molecules and bulk materials. Under the influence of an external source of electromagnetic radiation, quantum dots can be made fluorescent and thus determine their precise position using readily available optical techniques. Circular microneedle fixation patches with ammunition-shaped needles and cross with PLGA (Poly-DL-glycolic lactic acid, 0.8 cm in diameter with a 2 mm edge) were constructed. The patches were coated with quantum dots by placing 100 μL of CdSe / ZnS quantum dots (200 picoMolar, Invitrogen Qtracker ™ 655 nm) on the microneedles and air-dried. The arrays were examined with fluorescence using confocal microscopy.

The arrangements showed red fluorescence in the needles in the form of ammunition and cross indicating coating by the quantum dots. As shown in Figure 7, the coverage was shown at the top of the needles and under the sides towards the base. The cross-shaped needles demonstrated a more confluent coverage of the quantum dots, as shown in Figure 8. Absorption of quantum dots by lymphocytes can be observed through in vitro studies on cultured cells and in vivo studies on hairless mouse models.

EXAMPLE 10 Coating of insulin nanoparticles in microneedle arrays To demonstrate the efficiency of patch loads with nanoparticular biological molecules, a series of microneedle array patches were coated with nanostructured insulin. Insulin can be nanostructured using various methods that include supercritical fluid technologies. The size of the insulin particle is on average 300 nm. High-density PLGA circular patches and needle forms were coated with nanostructured insulin by placing 100 μL of nanostructured insulin in isoamyl alcohol (total 0.6 insulin units / patch) on the patches and air-dried. The patches were then examined for the presence of insulin using scanning electron microscope with field emission cannon (FEG-SEM), as shown in Figures 9 and 10. The patches demonstrated the presence of nanostructured insulin on both upper surfaces of the microneedles and under the side edges of the needles. The density of the insulin nanoparticles in the cross-shaped microneedles was smaller due to the larger surface area of the crosses compared to the ammunition.

EXAMPLE 11 Demonstration of skin penetration and delivery of quantum dots The ammunition-shaped patches were coated with quantum dots by placing 100 μL of CdSe / ZnS quantum dots (200 picoMolar in serum, Invitrogen Qtracker ™ 655nm) on the microneedles and dried on air. The patches were applied to the hind flank of nude mice by means of manual pressure. The patch was removed and the skin excised and examined with fluorescence using confocal microscopy, as shown in Figure 11. The skin showed red fluorescence on the surface of the stratum corneum indicating deposition of the quantum dot present at the base of the array. A deeper confocal image within the epidermis indicated red fluorescence in the form of an ammunition demonstrating the penetration of the microneedle to a total depth of approximately 60 μm, as shown in Figure 12. This experiment conclusively demonstrates that the arrangement of Microneedle can be used to deliver nanoparticles through a layer of the stratum corneum of the dermis.

EXAMPLE 12 Delivery of nanostructured insulin using arrangement microparches Preparation of insulin nanoparticles Insulin was nanostructured using a supercritical fluid process. An average particle size of 300 nm was obtained. The insulin was suspended in several solvents including isopropanol, isoamyl ethanol, ethanol, methanol or other coatings within the array. To coat the microarrays, insulin nanoparticles were suspended in solvent for a final concentration of 120 U / ml (4.32 mg / ml) and subjected to sonication for 60 seconds to ensure complete dispersion through the suspension. The suspension was then applied to each microarray (6U in 50 μl) and air drying was allowed. For subcutaneous delivery in the control experiments, the solution that was used to coat the microarrays was diluted 1: 300 in normal serum (final concentration 0.4U / ml).

Blood Glucose Experiments Hairless mice were anesthetized with pentobarbitone (60 mg / kg, Lp.). Blood samples were obtained by tail laceration and blood glucose was measured using a commercial glucose meter (Optimum ™ Xceed ™; Abbot Diagnostics). After obtaining two consecutive readings, the mice were treated as indicated and the blood glucose was recorded every 20 minutes for the recording of the experiment.

The mice were treated with a positive control (insulin suspension, lU / kg, s.c.), microarrays loaded with insulin (2 patches per mouse, 6U / patch), or negative control (12U of insulin applied directly to the skin without any microarray). The administration of insulin by means of a micropatch of arrangement can be observed in the mouse by a change in glucose in blood levels. Any analysis of the documents, records, materials, devices, articles or the like that have been included in the present specification are only for purposes of providing a context for the present invention. It is not considered an acknowledgment that any or all of these materials form part of the prior bases of the art or are of general common knowledge in the field pertaining to the present invention had they existed prior to the priority date of each of the claims of this application. It will be appreciated by persons skilled in the art that numerous variations and / or modifications to the present invention can be made as shown in the specific embodiments without departing from the spirit or scope of the present invention as broadly described. The present modalities are, therefore, to be considered in all respects as illustrative and not restrictive.

Claims

NOVELTY OF THE INVENTION Having described the present invention, it is considered as a novelty and, therefore, the content of the following is claimed as a priority: CLAIMS

1. - A device suitable for delivering at least one nanoparticle comprising a microneedle having at least one nanoparticle that is associated with at least part of a surface of the microneedle and / or at least part of the tissue of the microneedle.

2. The device according to claim 1, characterized in that the device has at least two microneedles.

3. The device according to claim 2, characterized in that the microneedles are arranged without pattern, arrangement or other such configuration.

4. The device according to any of the preceding claims, characterized in that the nanoparticle (s) is associated with at least a part of the outer surface of the microneedle.

5. The device according to any of the preceding claims, characterized in that the nanoparticle (s) is associated with pores on the surface of the microneedles.

6. The device according to any of the preceding claims, characterized in that the nanoparticle (s) is associated with at least a part of the tissue of the microneedle.

7. The device according to claim 1, characterized in that the nanoparticle (s) is associated with all the tissue of the microneedle.

8. The device according to claim 6, characterized in that the nanoparticle (s) is associated with internal pores in the tissue of the microneedle.

9. The device according to any of the preceding claims, characterized in that the association comprises a non-covalent interaction that is selected from any or any of the group comprising ionic bonds, hydrophobic interactions, hydrogen bonds, Van der Waals forces or dipole-dipole links.

10. The device according to any of claims 1 to 8, characterized in that the association is made by means of a covalent bond to a functional group in the microneedle.

11. The device according to claim 10, characterized in that the functional group (s) is (are) selected from the group comprising COOR5 CONR2, NH ?, SH, and OH, wherein R comprises H; organic or inorganic chain.

12. The device according to any of the preceding claims, characterized in that the microneedle (s) is manufactured (n) of a porous or non-porous material that is selected from the group comprising metals, natural or synthetic polymers, glasses, ceramics, or combinations of two or more of them.

13. The device according to claim 10, characterized in that the microneedle (s) is manufactured from a polymer selected from the group comprising: polyglycolic acid / polylactic acid, polycaprolactone, polyhydroxybutyrate-valerate, polyorthoester, and polyethylene oxide / polybutylene terephthalate, polyurethane, polymers of silicone, and polyethylene terephthalate, polyamine plus a trilayer of dextran sulfate, poly-L-lactic acid of high molecular weight, fibrin, methylmethacrylate (MMA) (hydrophobic, 70 mol% ) and 2-hydroxyethyl methacrylate - (HEMA) (hydrophilic 30 mol%), poly (ester-amide) elastomeric polymers (co-PEA), polyetheretherketone, (Peek-Optima), biocompatible thermoplastic polymer; conductive polymers, polystyrene or combinations of two or more thereof.

14. The device according to any of the preceding claims, characterized in that the microneedle (s) includes a layer or coating on at least a part of the surface of the microneedle (s) of a material electrically conductive

15. The device according to claim 14, characterized in that the electrically conductive material is selected from the group comprising conductive polymers; Compound conductive materials; doped polymers, conductive metallic materials or combinations of two or more thereof.

16. The device according to claim 15, characterized in that the conductive polymer is selected from the group comprising substitutable or irreplaceable polymers comprising polyaniline; polypyrrole; polysilicones; poly (3, -ethylenedioxythiophene); polymer doped with carbon nanotubes; polymers doped with metal nanoparticles, or combinations of two or more thereof.

17. The device according to any of claims 14 to 16, characterized in that the thickness of the layer or coating is from about 20 nm to about 20 μm.

18. - The device according to any of claims 14 to 17, characterized in that the electrically conductive material is layered or coated on the microneedle (s) by electrodeposition.

19. The device according to any of claims 14 to 18, characterized in that at least one nanoparticle is contained in the electrically conductive material.

20. The device according to any of the preceding claims, characterized in that the nanoparticle (s) is delivered to an organism and the microneedle (s) is made of a biocompatible material.

21. The device according to any of the preceding claims, characterized in that the microneedle (s) is not / are biodegradable (s).

22. The device according to any of the preceding claims, characterized in that the or each microneedle is solid.

23. The device according to any of the preceding claims, characterized in that the nanoparticle (s) is / are an active agent (s).

24. The device according to any of the preceding claims, characterized in that the nanoparticle (s) is / are a carrier (s).

25. The device according to claim 24, characterized in that the nanoparticle is associated with an active agent.

26. The device according to claim 25, characterized in that the active agent (s) is associated with the nanoparticle (s) by means of a covalent or non-covalent bond.

27. The device according to claim 25 or claim 26, characterized in that the nanoparticle encapsulates the active agent.

28. The device according to claim 25 or claim 26, characterized in that the active agent is incorporated in the nanoparticle (s).

29. The device according to any of claims 26 to 29, characterized in that the nanoparticle (s) is manufactured from a material that is selected from the group comprising metals, semiconductors, inorganic or organic polymers, magnetic colloidal materials, or combinations of two or more thereof.

30. The device according to claim 29, characterized in that the metal is selected from the group comprising gold, silver, nickel, copper, titanium, platinum, palladium and its oxides or combinations of two or more thereof.

31. The device according to claim 29, characterized in that the polymer is selected from the group comprising a conductive polymer; a hydrogel; agarose; polyglycolic acid / polylactic acid; polycaprolactone; polyhydroxybutyrate-valerate; poliortoester; polyethylene oxide / polybutylene terephthalate; polyurethane; polymeric silicon compounds; polyethylene terephthalate; polyamine plus a trilayer of dextran sulfate; high molecular weight poly-L-lactic acid; fibrin; copolymers of methyl methacrylate (MMA) and 2-hydroxyethyl methacrylate (HEMA), poly (ester-amide) elastomeric polymers (co-PEA); n-butyl cyanoacrylate; polyetheretherketone; (Peek-Optima), polystyrene or combinations of two or more of them.

32. The device according to claims 23 to 31, characterized in that the active agent is a biological agent.

33. The device according to claim 32, characterized in that the biological agent is a therapeutic and / or diagnostic agent.

34. The device according to claim 33, characterized in that the therapeutic agent is selected from the group comprising peptides, proteins, carbohydrates, nucleic acid molecules, an oligonucleotide fragment (s) or DNA or RNA, lipids, organic molecules , biologically active inorganic molecules or combinations of two or more thereof.

35.- The device according to claim 33, characterized in that the therapeutic agent is a vaccine.

36.- The device according to claim 35, characterized in that the vaccine is selected from the group comprising a vector containing a nucleic acid, oligonucleotide, expression gene as a vaccine or combinations of two or more thereof.

37.- The device according to claim 35, characterized in that the vaccine is selected from proteins or peptides as vaccines for diseases that are selected from the group comprising Johnes disease, bovine mastitis, meningococcal disease or combinations of two or more of the same.

38.- The device according to claim 37, characterized in that the vaccine comprises the Johne peptide disease that is selected from the group comprising: NVESQPGGQPNT (SEQ ID No I); QYTDHHSSLLGP (SEC ID No 2); LYRPSDSSLAGP (SEC ID No 3); and / or its variants.

39.- The device according to claim 37, wherein the vaccine comprises a peptide of bovine mastitis disease selected from the group comprising: MKKWFLILMLLGIFGCATQPSKVAAITGYDSDYYARYIDPDENKITFA IN VDGFVEGSNQEILIRGIHHVLTDQNQKIVTKAELLDAIRHQMVLLQLDY SYELVDFAPDAQLLTQDRRLLFANQNFEESVSLEDTIQEYLLKGHVILRK RVEEPITHPTETANIEYKVQFATKDGEFHPLPIFVDYGEKHIGEKLTSDEF RKIAEEKLLQLYPDYMIDQKEYTIIKHNSLGQLPRYYSYQDHFSYEIQ DR QRIMAKDPKSGKELGETQSIDNVFEKYLITKKSYKP (SEC ID No 4) ILIRGIHHVL (SEC ID No 5); IRHQMVLLQL (SEC ID No 6 ); and / or its variants.

40.- The device according to claim 33, characterized in that the diagnostic agent is a detectable agent.

41. The device according to claim 40, characterized in that the detectable agent is used in an analysis.

42. The device according to any of the preceding claims, characterized in that the outer diameter of the microneedle (s) is / are about 1 μm and about 100 μm.

43. - The device according to any of the preceding claims, characterized in that the length of the microneedle (s) is / are around 20 μm and 1 mm.

44. The device according to claim 43, characterized in that the length of the microneedle (s) is / are around 20 μm and 250 μm.

45.- The device according to any of the preceding claims, characterized in that the microneedle (s) is adapted (n) to provide an insertion depth of at least about 100 to 150 μm.

46.- The device according to any of the preceding claims, characterized in that the shape of the tip of the microneedle (s) is selected from the group comprising the square, circular, oval shapes, cross needle , triangular, chevron, serrated chevron, half moon or diamond.

47.- A method for manufacturing a device for the delivery of nanoparticles, the device comprising an array of microneedles and at least one nanoparticle that is associated with at least part of a surface of the microneedle, the method comprising the steps of: (i) covering at least a part of the surface of a microneedle array that is molded with the nanoparticles; (ii) molding the microneedles; characterized in that after leaving the mold, the nanoparticles are associated with the surface of the microneedles.

48. A method for manufacturing a device for the delivery of nanoparticles, the device comprising an array of microneedles and at least one nanoparticle that is associated with the pores on the surface of the microneedle, the method comprising the steps of: i) inducing porosity on at least a portion of the surface of the microneedles; ii) associate the nanoparticles with at least a part of the pores.

49. The method according to claim 48, characterized in that the step of inducing a porosity on the surface of the microneedles comprises the steps of: i) selectively leaching the micro or nanoparticles that are incorporated to the surface of the microneedle; ii) give physical, chemical or electrochemical treatment to the surface of the microneedles.

50.- A method for manufacturing a device for the delivery of nanoparticles, the device comprising an array of microneedles and at least one nanoparticle that is associated with at least part of the tissue of the microneedle, the method comprising the steps of : molding the microneedles in the presence of the nanoparticles; characterized in that after leaving the mold, the nanoparticles are associated with at least part of the tissue of the microneedles.

51.- A method for manufacturing a device for the delivery of nanoparticles, the device comprising an array of microneedles and at least one nanoparticle that is associated with at least a portion of the outer surface of the microneedle, the method comprising the steps of: i) functionalizing at least a portion of the outer surface of the microneedles with functional groups; ii) link the nanoparticles to the introduced functional groups.

52.- The method according to claim 51, characterized in that the step of functionalizing is selected from the group comprising oxidation, reduction, substitution, crosslinking, plasma, heat treatment or combinations of two or more thereof.

53. The method according to claim 52, characterized in that the functional group (s) introduced (s) are selected from the group comprising COOR, CONR2, NH2, SH, and OH, where R comprises H or an organic or inorganic chain.

54. The method according to any of claims 47 to 53, further comprising the step of coating at least a portion of the microneedles with an electrically conductive material.

55.- The method according to claim 54, characterized in that the electrically conductive material is selected from the group comprising conductive polymer; composite conductive material; doped polymer, metallic conductive materials or compounds thereof.

56. The method according to claim 55, characterized in that the conductive polymer is selected from the group of substitutable or irreplaceable polymers comprising polyaniline; polypyrrole; polysilicone; poly (3,4-ethylenedioxythiophene); polymers doped with carbon nanotubes; or polymers doped with metal nanoparticles.

57.- The method according to any of claims 54 to 56, characterized in that the thickness of the coating is from about 20 nm to about 20 μm.

58. The method according to any of claims 55 to 57, characterized in that the conductive polymer is coated on the microneedle by electrodeposition.

59.- A device suitable for delivering at least one agent comprising a microneedle that is made of an electrically conductive polymer and / or electrically conductive composite polymer, the microneedle having at least one agent that is associated with at least part of a surface of the microneedle and / or at least part of the tissue of the microneedle.

60.- The device according to claim 59, characterized in that the device has at least two microneedles.

61.- The device according to claim 60, characterized in that the microneedles are arranged in at least one arrangement.

62.- The device according to any of claims 59 to 61, characterized in that the agent (s) is associated with at least a part of the outer surface of the microneedle.

63.- The device according to any of claims 59 to 62, characterized in that the agent (s) is associated with pores on the surface of the microneedles.

64.- The device according to any of claims 59 to 63, characterized in that the agent (s) is associated with at least a part of the tissue of the microneedle.

65. - The device according to claim 64, characterized in that the agent (s) is associated with internal pores in the tissue of the microneedle. 66.- The device according to any of claims 59 to 65, characterized in that the association comprises a covalent or a non-covalent bond. 67.- The device according to claim 66, characterized in that the association is made by means of a covalent bond to a functional group in the microneedle. 68.- The device according to claim 67, characterized in that the functional group (s) is selected from the group comprising COOR, CONR2, NH2, SH, and OH, where R comprises H; organic or inorganic chain. 69.- The device according to any of claims 49 to 68, characterized in that the electrically conductive polymer is selected from the group of substitutable or irreplaceable polymers comprising polyaniline; polypyrrole; polysilicone; poly (3,4-ethylenedioxythiophene); polymer doped with carbon nanotubes; polymer doped with metal nanoparticle particles, or combinations of two or more thereof. The device according to any of claims 49 to 68, characterized in that the agent is selected from the group comprising biological agent, nanoparticle. 71. A microneedle comprising a plurality of removable biodegradable nanoparticles and / or a degradable nanoparticle. 72.- A method for delivering at least one nanoparticle (s) to a patient, the method including the steps of contacting at least one area of the patient with at least one microneedle that is associated with at least one nanoparticle, characterized in that at least one nanoparticle is delivered to the patient. 73.- A method according to claim 72, characterized in that the microneedle is in accordance with any of claims 1 to 45, and claims 59 to 70.