MXPA06009896A - Multi-component biological transport systems - Google Patents

Abstract

Compositions and methods are provided that are useful for the delivery, including transdermal delivery, of biologically active agents, such as non-protein non-nucleotide therapeutics and protein-based therapeutics excluding insulin, botulinum toxins, antibody fragments, and VEGF. The compositions and methods are particularly useful for topical delivery of antifungal agents and antigenic agents suitable for immunization. Alternately, the compositions can be prepared with components useful for targeting the delivery of the compositions as well as imaging components.

Description

SYSTEMS OF BIOLOGICAL TRANSPORT OF MULTIPLE COMPONENTS CROSS REFERENCE WITH RELATED APPLICATIONS This application is a continuation in part of the US Application No. 09 / 910,432 filed on July 20, 2001, which in turn claims the priority of the US Provisional Application Series No. 60 / 220,244, filed on July 21, 2000, the contents of which are incorporated in their entirety to the present invention as reference. DECLARATION WITH RESPECT TO THE RIGHTS FOR INVENTIONS PREPARED UNDER INVESTIGATION AND DEVELOPMENT SPONSORED BY THE FEDERATION Not applicable. Background of the Invention Gene delivery systems can be broadly classified into two groups: viral and non-viral. Viral systems have significant toxicity risks, and have resulted in significant complications and death in clinical trials. Non-viral systems are less efficient than viral methods, although they offer the potential of custom applications to increase specificity and potentially decrease toxicity. Non-viral strategies can be broadly classified as lipid-based and non-lipid based. The strategy presented in the present invention can be applied to any of the methods non-existing virals, as will be described in the present invention. The simplest non-viral system is the direct supply of DNA. Due to the negative charge of DNA, very little of the DNA actually enters the cell and most of it degrades. Virtually none of the DNA enters the nucleus without a sequence of nuclear direction in the strategy. Conventionally, another factor is used to increase the efficiency of the gene / product supply (DNA, RNA, or more recently protein therapeutics) either by mechanical effects such as electroporation, ultrasound, "gene gun" and direct microinjection or by neutralization of loading and chemical effects with agents such as calcium phosphate, polylysine and liposome preparations. In the latter strategies, charge neutralization has been shown to increase nonspecific efficiencies several times with respect to even the chemical / mechanical effects of liposome preparations alone. Based on these results and similar results, it has been concluded that DNA and RNA require charge neutralization for efficiency in cellular uptake, since the negative charge of the DNA essentially excludes transport except by means of endolysis with subsequent lysosome fusion ( that escapes with the addition of other agents). Most transfection agents actually use an excess of positive charge in proportions of 2 to 4 times with respect to the net negative charge of DNA. The resulting positive hybrid binds surface proteoglycans in ionic form charged negatively and dramatically increases the subsequent uptake. Certain transfection agents appear to have a cellular tropism, most of which is also due to the steric and charge patterns which most effectively direct the particular proteoglycans, which vary in specific patterns of the cell type. Even with suitable agents (ie, correct tropism), charge neutralization alone in combination with liposomes remains extremely inefficient in relation to viral strategies. Therefore, the community has identified a number of peptides and peptide fragments that facilitate the efficient entry of a complex into a cell and go through any endolysosome stage. Several transport factors allow even an efficient nuclear entry. In a process, the role of transport is directly linked to the therapeutic product of interest (small drug, gene, protein, etc.). This method requires that a new drug be produced adhered to the transport factor, be purified and tested. In many cases, these hybrids will actually constitute new drugs and will require a complete testing process. This process results in additional risk and significant expenses. As an alternative, a number of strategies merely employ the mixing of the agent in non-specific (or even surface-specific) form in the liposome preparations in the form of carriers for a drug / DNA / factor. Although there is an improvement with respect to direct or simpler modalities in In terms of efficiencies, these methods remain inefficient (relatively relative to the virus) and considerably more toxic than simple non-viral strategies. Part of this inefficiency is due to a poor nuclear translocation. As a result, strategies include adding nuclear translocation signals to the complex detailed above, either as part of the therapeutic factor hybrid or as part of the liposome mixture. Additional refinements have included efforts to reduce DNA / RNA / factor degradation. Possibly the most important refinements in the basic strategies presented above have included specific ligands or other steering agents along with the therapeutic factor. These strategies offer the potential for highly reduced non-specific toxicity and substantial improvements in efficiency, particularly when combined with the efficiency agents described above. However, normal strategies depend on covalent linkages to a single transporter and therefore need a specific synthesis (to ensure that spherical conditions in a degree of substitution scheme do not favor a single factor over others - that is, for ensure that each efficiency factor and each image generation portion and each address portion is in the skeleton). This makes a number of specific constructs virtually impossible (for example, X fragment of sialyl-Lewis and a Fab fragment for a surface antigen, since the Spherical limitations can avoid efficient linking of one or the other in most schemes, and in turn can interfere with efficiency factors). Although promising in concept, these methods represent costly solutions, with very low performance (in terms of synthesis), and unproven for this problem. As it should be evident, with each stage of development in the supply of gene and non-viral factor, problems have been found, and in the next stage, they have been solved with a degree of added complexity. Each improvement represented an additional step with respect to the previous standard. However, the added complexity carries the risk from the point of view of patient care and inefficiency and cost from a production point of view. These barriers have led to a greatly reduced enthusiasm for these potentially promising therapies in another way. What is needed are new methods and compositions that are broadly applicable to compositions of various therapeutic and cosmetic agents that can be targeted or image generated to maximize delivery to a particular site. Surprisingly, the present invention provides such compositions and methods. The present invention further relates to formulations for the transdermal delivery of proteins such as insulin, and also of larger therapeutic and diagnostic substances, for example, substances having a molecular weight of 50,000 and which also include proteins such as botulinum toxin and other biologically active agents such as, for example, insulin, botulinum toxin, a therapeutic protein that does not alter in blood therapeutic levels, a nucleic acid-based agent, a therapeutic agent without nucleic acid without protein such as certain antifungals or alternatively an agent for immunization. The present invention specifically excludes antibody fragments that have no biological activity other than the binding of a specific antigen only when the term "therapeutic" or "biologically active protein" is used. Since the antigens suitable for immunization have other biological activities such as assembly of an immune response, they remain included in the appropriate aspects of the present invention. In addition, agents that have a biological activity or therapeutic effect by binding a specific antigenthus blocking the ligand linkage or altering the conformation of the antigen are included in the present invention. Botulinum toxins (also known as botulinum toxins or botulinum neurotoxins) are neurotoxins produced by Clostridium botulinum of gram-positive bacteria. They act to produce muscle paralysis by preventing synoptic transmission or release of acetylcholine through the neuromuscular junction, and are considered to act in other forms as well. Its action essentially blocks the signals that can normally cause muscle spasms or contractions, giving as result paralysis or could cause glandular secretions or overexcretion such as hyperhidrosis or acne. Botulinum toxin is classified into eight neurotoxins that are serologically related, although they are different. Of these, seven can cause paralysis, namely the botulinum neurotoxin of serotypes A, B, C, D, E, F, and G. Each of these is distinguished by neutralization with type-specific antibodies. Each type can be recombinant of natural emergence in production or constructed variants such as protein fusions. However, the molecular weight of the botulinum toxin protein molecule, for all seven of these active botulinum toxin serotypes that naturally occur in their recombinant forms, is approximately 150 kD. As released through the bacterium, the botulinum toxins are complexes comprising the 150 kD botulinum toxin protein molecule in question, along with associated toxin-free proteins. The botulinum toxin type A complex can be produced by Clostridia bacterium as forms of 900 kD, 500 kD, and 300 kD. Botulinum toxin types B and C are apparently produced only as a complex of 700 kD or 500 kD. Botulinum toxin type D is produced as complexes of both 300 kD and 500 kD. Botulinum toxin type E and F are produced only as complexes of approximately 300 kD. Complexes (ie, molecular weight greater than about 150 kD) are considered to contain a hemagglutinin protein without toxin and a protein without non-toxin and non-toxic haemagglutinin. These two toxin-free proteins (which together with the botulinum toxin molecule comprises the relevant neurotoxin complex) can act to provide stability against denaturation for the botulinum toxin molecule and protection against digestive acids when the toxin is ingested. In addition, it is possible that the larger botulinum toxin complexes (greater than a molecular weight of approximately 150 kD) may result in a slower diffusion range of the botulinum toxin outside the intramuscular injection site of a botulinum toxin complex. . The different serotypes of botulinum toxin vary in the animal species that affect them and in the severity and duration of the paralysis they cause. For example, it has been determined that botulinum toxin type A is 500 times more potent, as measured by the range of paralysis produced in the rat, than botulinum toxin type B. In addition, botulinum toxin type B has been determined for non-toxic primates at a dose of 480 U / kg, approximately 12 times the LD50 of the primate for type A. Due to the molecule size and molecular structure of the botulinum toxin, it can not cross the stratum corneum and the multiple layers of the architecture of the underlying skin. Botulinum toxin type A, is said to be the most lethal natural biological agent known to man. The spores of C. botulinum, are found in the soil and can grow in sterilized and improperly sealed food containers. Ingestion of the bacteria can cause botulism, which which can be fatal. At the same time, the muscle paralyzing effects of botulinum toxin have been used for therapeutic purposes. The controlled administration of botulinum toxin has been used to provide muscle paralysis to treat conditions, for example, neuromuscular disorders characterized by hyperactive skeletal muscles. Conditions that have been treated with botulinum toxin include hemifacial spasm, generation spasmodic torticollis in adults, anal fissure, blepharospasm, cerebral palsy, cervical dystonia, migraine headaches, strabismus, temperomandibular joint disease, and various types of constrictions and spasms. Muscular More recently, the effects of muscle paralysis of botulinum toxin, have been convenient in therapeutic and cosmetic facial applications, such as treatment of wrinkles, fine lines and other results of spasms or contractions of facial muscles. Botulism, a symptom complex characteristic of exposure to systemic botulinum toxin, has existed in Europe since ancient times. In 1895, Emile P. van Ermengem first isolated the bacillus that forms the anaerobic spore of unprocessed salted pork obtained from post-mortem tissue of victims who died of botulism in Belgium. Van Ermengem discovered that the disease was caused by an extracellular toxin that was produced by what he called "Bacillus botulinus (Van Ermengem, Z. Hyyg Infektionskr, 26: 1-56, Rev. Infecí. (1987)). name was changed in 1922 to Clostridium botulinum. The name Clostridium was used to reflect the anaerobic nature of the microorganism and also its morphological characteristics (Carruthers and Carruthers, Can J. Ophthalmol, 31: 389-400 (1996)). In the 1920s, a crude form of botulinum toxin type A was isolated after further eruptions of food poisoning. Dr. Herman Sommer of the University of California, San Francisco, made the first attempts to purify the neurotoxin (Borodic and Associates, Ophthalmic Plast Recostr. Surg., 7: 54-60 (1991)). In 1946, Dr. Edward J. Schantz and his colleagues isolated the neurotoxin in crystalline form (Schantz and Associates, In: Jankovi J., Hallet M. (Eds) Therapy with Botulinum Toxin, New York, NY: Marcel Dekker, 41 -49 (1994)). In 1949, Burgen and his associates had the ability to show that botulinum toxin blocks impulses through the neuromuscular junction (Burgen and Associates, J. Physiol., 109: 10-24 (1949)). Allan B. Scott first used botulinum toxin A (BTX-A) in monkeys in 1973. Scott demonstrated reversible ocular muscle palsy lasting three months (Lamanna, Science, 130: 763-772 (1959)). Soon after, it was reported that BTX-A is a successful treatment in humans for strabismus, blepharospasm and spasmodic torticollis (Baron and Associates, In: Baron EJ, Peterson LR, Finegold SM (Eds), Bailey &Scotts Diagnostic Microbiology, St. Louis, MO: Mosby Year Book, 504-523 (1994), Carruthers and Carruthers, Adv. Dermatol, 12: 325-348 (1997), Markowitz, En: Strickiand GT (Eds.) Tropical Hunters Medicine, 7th Edition, Philadelphia: W. B. Saunders, 441 -444 (1991)). In 1986, Jean and Alastair Carruthers, a couple of spouses, where said team consists of an ocuplastic surgeon and a dermatologist, began to involve the cosmetic use of botulinum toxin-A (BTX-A) for the treatment of wrinkles associated with movement in the area of glabella (Schantz and Scott, In Lewis GE (Ed) Biomedical Aspects of Botulinum, New York: Academic Press, 143-150 (1981)). The use of Carruthers by BTX-A for the treatment of wrinkles, led to its seminal publication of this method in 1992 (Schantz and Scott, In Lewis GE (Ed) Biomedical Aspects of Botulinum, New York: Academic Press, 143-150 ( 1981 )). In 1994, the same type reported experiences with other wrinkles associated with movement in the face (Scott, Ophthalmol, 87: 1044-1049 (1990)). This, in turn, led to the birth of the era of cosmetic BTX-A treatment. The skin protects the body's organs from external environmental damage and acts as a thermostat to maintain body temperature. It consists of different layers, each with specialized functions. The major layers include the epidermis, the dermis and the hypodermis. The epidermis is a stratification layer of epithelial cells that underlies the dermis, which consists of connective tissue. Both the dermis and the epidermis are additionally supported by the hypodermis, and the inner layer of adipose tissue. The epidermis, the most superior layer of the skin, has a unique 0.1 to 1.5 mm thickness (Inlander, Skin, New York, NY: People's Medical Society, 1-7 (1 998)). It consists of keratinocytes and is divided into several layers based on their state of differentiation. The epidermis can be further classified into the stratum corneum and the viable epidermis, which consists of granular melfigian and basal cells. The stratum corneum is hygroscopic and requires at least 10% moisture by weight to maintain its flexibility and softness. The hygroscopicity is attributed in part to the ability to hold water from keratin. When the horny layer loses its softness and flexibility, it becomes rough and stiff, resulting in dry skin. The dermis, which lies just below the epidermis, is 1.5 to 4 millimeters thick. It is the thickest of the three layers of the skin. In addition, the dermis is also the place for most skin structures, including sweet and oily glands (which secrete substances through the openings in the skin and the so-called pores or comedones), hair follicles, nerve endings and blood and lymphatic vessels (I nlander, Skin, New York, NY: People's Medical Society, 1-7 (1,998)). However, the main ones of the dermis are collagen and elastin. The epidermis is the deepest layer of the skin. It acts as both an insulator for the preservation of body heat, and an impact absorber for the protection of organs (I nlander, Skin, New York, NY: People's Medical Society, 1-7 (1 998)). In addition, the hypodermis also stores fat for energy reserves. The pH of the skin is normally between 5 and 6. This acidity is due to the presence of amphoteric amino acids, lactic acid and fatty acids from the secretions of the sebaceous glands. The term "acid mantle" refers to the presence of water-soluble substances in most regions of the skin. The regulation capacity of the skin is due in part to these secretions stored in the corneal layer of the skin. Wrinkles, one of the signs that reveal age, can be caused by biochemical, histological and physiological changes that accumulate from environmental damage (Benedetto, International Journal of Dermatology, 38: 641-655 (1999)). In addition, there are other secondary factors that can cause the folds, grooves, and crises characteristic of facial wrinkles (Stegman and Associates, The Skin of the Aging Face Cosmetic Dermatological Surgery, 2nd Edition, St. Louis, MO: Mosby Year Book: 5-15 (1990)). These secondary factors include the constant action of gravity, Ipartido in two of frequent and constant position on the skin (for example, during sleep) and repeated facial movements caused by the contraction of facial muscles (Stegman and Associates, The Skin of the Aging Face Cosmetic Dermatological! Surgery, 2nd Edition, St. Louis, MO: Mosby Year Book: 5-15 (1990)). Different techniques have been used in order to potentially modify some of the signs of aging. These techniques range from facial moisturizers containing hydroxy-alpha and retinol acids to surgical procedures and injections of neurotoxins.

One of the main functions of the skin is to provide a barrier to the transport of water and potentially harmful substances for normal homeostasis. The body could dehydrate quickly without a resistant, semi-permeable skin. The skin helps prevent the entry of substances dangerous to the body. Although most substances can not penetrate the barrier, a number of strategies have been developed to selectively increase skin permeability with varying success. Since BTX can not penetrate the skin efficiently, in order to provide the therapeutic effects of BTX, the toxin must be injected normally into the skin. The Federal Food and Drug Administration has tested this procedure for the treatment of wrinkles, and BTX products are now marketed for this treatment. In such treatments, the botulinum toxin is administered by controlled injection or carefully monitored, creating large deposits of toxins at the treatment site. However, such treatment can not be uncomfortable and usually involves some pain. Topical application of botulinum toxin provides a safer and more desirable treatment alternative due to the painless nature of the application, the larger treatment surface area that will be covered, the ability to formulate a pure toxin with superior specific activity. , reduced training to apply the botulinum therapeutics, smaller doses needed for the effect, and no large deposits of toxins in order to achieve a clinical therapeutic result. Transdermal administration of other therapeutics is also an area of interest, due, for example, to the potential for decreased discomfort of the patient, direct administration of the therapeutic agents in the bloodstream and opportunities for a monitored supply through the use of devices and / or formulations and controlled release techniques. A substance for which the ease of administration is desired is that of insulin, which in many cases must be administered by injection (including self-injection). The ease of administration could be convenient for larger proteins, such as botulinum toxin. Other agents that do not easily cross the skin, but are substantially smaller than insulin or have different physiochemical properties, and therefore very different ranges and capacities to traverse the skin with and without additional mials facilitthis transfer. The additional interaction of each of the mials, to facilitthe transfer, is unique to each one. Brief Description of the Invention In one aspect, the present invention provides the composition comprising a non-covalent complex of: a) a skeleton positively charged; and b) at least two members selected from the group consisting of: i) a first skeleton loaded in a negative form that has a plurality of adhered image generating portions, or alternatively a plurality of negatively charged image generating portions; ii) a second negatively charged skeleton having a plurality of adhered steering agents, or alternatively, a plurality of negatively charged steering portions; iii) at least one member selected from RNA, DNA, ribozymes, modified oligonucleic acids and cDNAs encoding a selected transgene; iv) DNA encoding at least one persistence factor; v) a third skeleton loaded in a negative form having a plurality of biological agents adhered, or a biological agent loaded in a negative form; where the complex carries a net positive charge and at least one of the members is selected from i), ii), iii), or v). The biological agent, in this aspect of the present invention, can be either a therapeutic agent or a cosmetological agent. The present invention specifically excludes antibody fragments that have no biological activity other than only binding of a specific antigen when the term "therapeutic" or "biologically active protein" is used. Since the antigens suitable for immunization have other biological activities such as assembly of an immune response, these remain included in the appropriate aspects of the present invention. In addition, agents that have a biological activity in therapeutic effect by binding a specific antigen, thereby blocking the ligand binding or altering the conformation of the antigen, are included in the present invention. Alternatively, the candidate agents can be used to determine the efficacy in vivo in these non-covalent complexes. In another aspect, the present invention provides a composition comprising a non-covalent complex of a positively charged backbone having at least one adherent efficiency group and at least one nucleic acid member selected from the group consisting of RNA, DNA, ribozymes, modified oligonucleic acids and cDNA encoding a selected transgene. In another aspect, the present invention provides a method for delivering a biological agent to a cell surface in a subject, wherein the method comprises administering to the subject a composition as described above. In yet another aspect, the present invention provides a method for preparing a pharmaceutical or cosmetological composition, wherein the method comprises combining a positively charged skeletal component and at least two members selected from the group consisting of: i) a first skeleton negatively charged having a plurality of adhered image generating portions, or alternatively, a plurality of generating portions thereof image loaded in a negative way; ii) a second negatively charged skeleton having a plurality of adhered steering agents, or alternatively, a plurality of negatively charged steering portions; iii) at least one selected member of RNA, DNA, ribozymes, modified oligonucleic acids and cDNA encoding a selected transgene; iv) DNA encoding at least one persistence factor; and v) a third skeleton loaded in a negative form having a plurality of biological agents or cosmetological agents adhered, or a biological agent or a cosmetological agent negatively charged; with a pharmaceutically or cosmetologically acceptable carrier to form a non-covalent complex having a net positive charge, provided that at least one of the members is selected from i), ii), iii), or v). In yet another aspect, the present invention provides a kit for formulating a pharmaceutical or cosmetological delivery composition, wherein the kit comprises a positively charged skeleton component and at least two components selected from groups i) through v) above, along with instructions to prepare the supply composition. In still another aspect, the present invention relates to a composition comprising a biologically active agent, such as insulin, botulinum toxin, and other proteins that do not alter in a therapeutic way the blood glucose levels, a nucleic acid-based agent, a therapeutic agent without nucleic acid without protein, such as antifungal agents or alternatively, an agent for immunization , and a vehicle comprising a positively charged vehicle having a branched skeleton loaded in positively bonded form or "efficiency groups", all described in the present invention. The present invention specifically excludes fragments of antibodies that have no biological activity other than solely in the form of a specific antigen when the term "therapeutic" or "biologically active protein" is used. Since the antigens suitable for immunization have other biological activities, such as assembly of an immune response, these remain in the proper aspects of the present invention. In addition, agents that have a biological activity or a therapeutic effect by binding a specific antigen, thereby blocking the ligand linkage or altering the conformation of the antigen, are included in the present invention. The biologically active agent is preferably insulin, botulinum toxin (BTX), an antigen for immunization, or certain antifungal agents. Suitable antifungal agents include, for example, amphotericin B, fluconazole, flucytosine, itraconazole, ketoconazole, clotrimazole, econozole, griseofulvin, miconazole, nystatin, cyclopirox, and the like. More preferably, the positively charged vehicle is a polypeptide loaded in the form positive short-chain or comparatively medium or a polymer without peptidyl positively charged, for example, a polyalkyleneimine. When the biologically active agent is botulinum toxin, the present invention further relates to a method for producing a biological effect, such as muscle paralysis, reduction of hypersecretion, or exudation, treatment of neurological pain or migraine headache, reduction of spasms. muscles, prevention or reduction of acne, or reduction or increase of immune response, applying topically a composition containing an effective amount of botulinum toxin, preferably to the skin, of a subject or patient in need of such treatment. The present invention also relates to a method for producing an aesthetic and / or cosmetic effect, for example by topically applying botulinum toxin to the face instead of by injection into the facial muscles. When the biologically active agent is insulin, the present invention relates to a method for transdermally delivering insulin to a subject by applying to the skin or epithelium of the subject an effective amount of said insulin-containing composition, or a combination of insulin and insulin. skeleton charged positively. Proteins that normally do not have the ability to traverse the skin or epithelium are appreciably related to the complex of the same agent and vehicles of the present invention and which have no therapeutic effect on the blood glucose decrease, have surface properties and physiochemicals very different from insulin, which normally it may be certain that the technique for producing the transdermal insulin supply may have positive results for any other proteins. However, vehicles of the present invention having a positively charged skeleton with positively charged branching groups, as described in the present invention, have a very surprising ability to provide transdermal delivery of other such proteins. , including, for example, botulinum toxin. Particular vehicles suitable for the transdermal delivery of particular proteins can be easily identified using tests such as those described in the examples. Said protein can, for example, be a large protein having a molecular weight above 50,000 kD or below 200,000 kD. As used in the present invention, the word "therapeutic" within the context of blood glucose, refers to a decrease in blood glucose levels sufficient to alleviate acute symptoms or signs of hyperglycemia, for example in diabetic patients. In all aspects of the present invention, the association between the carrier and the biologically active agent is by non-covalent interaction, which may include, for example, ionic interactions, hydrogen bonding, van der Waals forces, or combinations of same. In certain aspects of the present invention, the transdermal delivery of therapeutic proteins with the ability to achieve therapeutic alterations of blood glucose, are excluded in specific form. As used in the present invention, antigenic agents suitable for immunization can be protein-based antigens that do not therapeutically alter blood glucose levels, agents without nucleic acid, without protein or hybrids thereof. The nucleic acids encoding the antigens are specifically unsuitable for the compositions of the present invention. Therefore, the included agents are the proper antigens for immunization. Suitable antigens include, for example, those for environmental, pathogenic or biohazard agents. Suitable agents preferably include, for example, antigens related to botulism, malaria, rabies, anthrax, tuberculosis, or related to childhood immunizations such as hepatitis B, diphtheria, pertussis, tetanus, influenza to Haemophilus type b, inactivated poliovirus, measles, mumps , rubella, chicken pox, pneumococcus, hepatitis A, and influenza. The positively charged vehicles or skeletons with their branching groups positively charged, as described in the present invention, are themselves novel compounds and form another aspect of the present invention. The present invention also provides a method for preparing a pharmaceutical or cosmetic composition comprising combining a carrier comprising a positively charged polypeptide or a positively charged peptidyl-free polymer such as a long-chain polyalkyleneimine, having the polypeptide polymer or polymer without peptidyl branching charged in positive form or "efficiency" groups as defined in the present invention, with a biologically active agent such as, for example, insulin, botulinum toxin, a therapeutic protein that does not alter therapeutically blood glucose levels, a nucleic acid-based agent, a therapeutic agent without nucleic acid, without protein such as antifungal agents or alternatively, an agent for immunization. The present invention also provides a kit for preparing or formulating said composition comprising the carrier and the therapeutic substance, as well as additional batches that are needed to produce a usable formulation, or a premix which in turn can be used to produce said formulation. . Said equipment may consist of an applicator or other apparatus for applications of the compositions or components thereof and methods of the present invention. As used in the present invention, "apparatuses" can refer, for example, to an instrument or applicator for delivery or to another preparation technique, to form or apply the compositions and methods of the present invention. The present invention also comprises apparatuses for transdermal transmission of a biologically active agent such as, for example, insulin, botulinum toxin, a therapeutic protein that alters in therapeutic form the levels of glucose in the blood, a nucleic acid-based agent, a therapeutic agent without nucleic acid, without protein such as certain antifungals, or alternatively, an agent for immunization that is contained within a composition, which in turn, in one embodiment, comprises a vehicle comprising a positively charged polypeptide preferably with a short to intermediate chain length or a polymeric carrier without longer chain peptidyl having positively charged branching or "efficiency" groups, as defined in the present invention, and a therapeutic agent such as those mentioned above . Such devices may be as simple in construction as a skin patch, or may be a more complicated apparatus that includes means for supplying and monitoring the supply of the composition, and optionally means for monitoring the condition of the subject in one or more aspects, including monitoring of the reaction of the subject to the substances that are being supplied. In all aspects of the present invention, the association between the vehicle and the biologically active agent is by non-covalent interaction, which may include, for example, ionic interactions, hydrogen bonding, Van der Waals strength or combinations thereof. Alternatively, the apparatus may contain only the therapeutic biologically active agent, for example, insulin, botulinum toxin, a therapeutic protein that does not alter in blood therapeutic levels, a nucleic acid-based agent, a therapeutic agent without nucleic acid, without protein such as certain antifungals or alternatively, an agent for immunization, and the vehicle can be applied separately to the skin. Accordingly, the present invention also comprises equipment that includes both an apparatus for delivery through the skin and a material containing the positively charged vehicle or skeleton, and which is suitable for application to the skin or epithelium of a subject . In general, the present invention also comprises a method for administering a biologically active agent such as, for example, insulin, botulinum toxin, a therapeutic protein that does not alter in a therapeutic manner the levels of glucose in the blood, an acid-based agent. nucleic acid, a therapeutic agent without nucleic acid, without protein such as certain antifungals or alternatively, an agent for immunization to a subject or patient in need thereof, comprising the administration in topical form of an effective amount of said biologically active agent together with a polypeptide polymer or without positively charged polypeptidyl, such as a polyalkyleneimine having positively charged branching groups, as described in the present invention. By the term "together with" is meant that the two components - biologically active agent and positively charged vehicle - are administered in a combination procedure, which may involve either combining them into a composition, which is subsequently administered to the subject , or manage them separately, although in a way that they act together to provide the supply of requirements of an effective amount of the biologically active agent. For example, a composition containing the positively charged vehicle can first be applied to the skin of the subject, followed by the application of a skin patch or other apparatus containing the biologically active agent. The present invention relates to methods for applying biologically active agents such as, for example, insulin, botulinum toxin, a therapeutic protein that does not alter in blood therapeutic levels, a nucleic acid-based agent, an agent therapeutic without nucleic acid without protein such as certain antifungals or alternatively, an agent for immunization as defined in the present invention, epithelial cells, including those other than skin epithelial cells, for example, epithelial or epithelial cells of the gastrointestinal system. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 provides a schematic representation of the components used in the present invention. Figure 2 provides a schematic representation of various embodiments of the present invention. Figures 3 and 4 represent the results of the transdermal delivery of a plasmid containing the transgene for E. coli beta-galactosidase, as described in example 2. Figure 5 represents the results of the transdermal delivery of a plasmid which contains the transgene for beta- E. coli galactosidase as described in Example 3. Figure 6 represents the results of the transdermal delivery of a plasmid containing the transgene for E. coli beta-galactosidase as described in example 4. The figure 7, represents the results of the transdermal delivery of a botulinum toxin as described in example 5. Figure 8 is a photographic illustration of the results of the transdermal delivery of a botulinum toxin as described in example 6. The figure 9, is a photographic illustration of how in the generation of images the complexes of example 9 follow the bright field distribution (panels a and c) for melanoma pigmented cells with fluorescent optical imaging agents (b and d panels) for two different fields and different magnifications (panels a and b in 1 0X versus panels c and d in 40X of magnifications). Detailed Description of the General Invention The present invention provides a component-based system for the selective, persistent delivery of imaging agents, genes or other therapeutic agents. The individual characteristics of the compositions can be selected by designating the desired components in additional formulations. In addition, in one aspect, portions of Specific image generation and direction are provided in skeletons charged negatively or separately, which will form a non-covalent ion complex with a positive skeleton. By placing these components in a negatively charged skeleton, the present invention eliminates the need to adhere components to precise locations in a positive skeleton, as employed in other strategies (increasing complexity and expense and decreasing efficiency to such a level). that a successful combination has not been reported due to spherical limitations In another aspect, certain substances can be administered transdermally through the use of certain positively charged vehicles alone, without requiring the inclusion of a negative skeleton. , the substance still derived therefrom has sufficient negative charge to associate with the positively charged vehicles of the present invention, in non-covalent form.The term "efficient" within this context, refers to an association that can be determined , for example, through the change in particle size or spectrophotometry untional versus single components. A further understanding of the present invention is provided with reference to Figure 1. In this figure, the components are shown as (1) a solid skeleton that has positively charged groups attached (also referred to as efficiency groups shown as dark circles attached to an obscured bar), for example (Gly) n? - ( Arg) n2 (in wherein the subscript n1 is an integer from 3 to about 5, and the subscript n2 is an integer non of from about 7 to about 17) or TAT domains; (2) a short skeleton loaded in a negative form that has adhered portions of image generation (open triangles attached to a clear bar); (3) a short skeleton loaded in a negative form having adhered steering agents and / or therapeutic agents (open circles adhered to a clear bar); (4) an oligonucleic acid, RNA, DNA or cDNA (bar with clear transverse stripes); and (5) DNA encoding persistence factors (bar with dark transverse stripes). Figure 2 illustrates several examples of multi-component compositions, wherein the groups are illustrated as a group as set forth in Figure 1. For example, in Figure 2, a first multi-component composition in which a skeleton is illustrated is illustrated. positively charged has associated an image generation component, an address component, a nucleic acid and a persistence factor. A second multi-component composition is illustrated, which is designed for diagnostic and / or prognostic image generation. In this composition, the positively charged skeleton is made up of compounds with both imaging components and steering components. Finally, a third multi-component system is illustrated which is useful for gene delivery. In this system, a complex is formed between a positively charged skeleton, an address component, a gene of interest and a DNA that codes for a persistence factor. The present invention, which is described in more detail below, provides the number of additional compositions useful in therapeutic and diagnostic programs. Description of the Modalities Compositions In light of the foregoing, the present invention provides in one aspect, a composition comprising a non-covalent complex of: a) a skeleton positively charged; and b) at least two members selected from the group consisting of: i) a first negatively charged skeleton having a plurality of adhered image generating portions; or alternatively, a plurality of negatively charged image generation portions; ii) a second negatively charged skeleton having a plurality of adhered steering agents; or alternatively, a plurality of negatively charged address portions; iii) at least one member selected from RNA, DNA, ribozymes, modified oligonucleotide acids and cDNA encoding an unselected transgene; iv) DNA encoding at least one persistence factor; and v) a third skeleton loaded in a negative form having a plurality of biological agents adhered, or a biological agent loaded in a negative form; where the complex carries a net positive charge and at least one of the members is selected from i), ii), iii), or v). In a group of modalities, the composition comprises at least three members selected from groups i) to v). In another set of modalities, the composition comprises at least one member from each of groups i), ii), iii), and iv). In yet another group of modalities, the composition comprises at least one member from each of the groups i) to i). In another group of modalities, the composition comprises at least one member from each of groups ii), iii), and iv). Preferably, the positively charged skeleton has a length of from about 1 to 4 times the combined lengths of the members of group b). Alternatively, the positively charged skeleton has a loading ratio of from about 1 to 4 times the combined charge of the members of group b). In some embodiments, the charge density is uniform and the proportions of length and charge are approximately the same. The proportions of size to size (length) can be determined based on molecular studies of the components, or can be determined from the masses of the components. By the term "negatively charged" it is understood that the The vehicle has a positive charge under at least some solution phase conditions, more preferably at least under some physiologically compatible conditions. More specifically, the term "positively charged" as used in the present invention means that the group in question contains functionalities that are charged under all pH conditions, such as a quaternary amine, or that contain a functionality which can acquire positive charge under certain phase-solution conditions, such as pH changes in the case of primary amines. More preferably, the term "positively charged" as used in the present invention, refers to all those having the behavior of associating with anions through physiologically compatible conditions. Polymers with a multiplicity of positively charged portions do not need to be homopolymers, as will be appreciated by those skilled in the art. Other examples of positively charged portions are well known in the prior art and can be used easily, as will be appreciated by those skilled in the art. The positively charged vehicles described in the present invention, which by themselves do not have a therapeutic activity, are novel compounds having utility, for example in compositions and methods as described herein. Therefore, in another aspect of the present invention, we will detail these novel compounds that include any vehicle comprising a skeleton loaded in the form positive that positively charged branching groups adhere to, as described in the present invention, and that do not themselves have a therapeutic biological activity. The present invention specifically excludes antibody fragments that have no biological activity, other than only binding a specific antigen when the term "therapeutic" or "biologically active protein" is used. Since the antigens suitable for immunization have other biological activities, such as assembly of an immune response, these remain included in the appropriate aspects of the present invention. In addition, agents that have a biological activity or a therapeutic effect through the binding of a specific antigen, thereby blocking the ligand binding or altering the conformation of the antigen, are included in the present invention. In another modality, the present invention provides in one aspect a composition comprising a biologically active agent such as, for example, insulin, botulinum toxin, a therapeutic protein that does not alter in therapeutic form the levels of glucose in the blood, an acid-based agent. nucleic acid, a therapeutic agent without nucleic acid, without protein such as certain antifungals, or alternatively, an agent for immunization and a carrier comprising a positively charged backbone, for example a polypeptide polymer or without peptidyl positively charged which can be any of a hetero- or homopolymer, such as a polyalkyleneimine, having the polypeptide polymer or without peptidyl branching or "efficiency" groups positively charged, as defined in the present invention. Each therapeutic without protein, without nucleic acid and with a protein-based therapeutic have different physiochemical properties that alter the total characteristics of the complex. Said positively charged vehicles are among the materials described above as positively charged skeletons. The present invention also provides a method for administering a therapeutically effective amount of a biologically active agent as mentioned in the present invention, which comprises applying to the skin an epithelium of the subject (which may be a human or other mammal). The biologically active agent and a positively charged skeletal amount having branching groups that are effective to provide transdermal delivery of the biologically active agent to the subject. In such a method, the biologically active agent and the positively charged vehicle can be applied as a previously mixed composition, or they can be applied separately to the skin or epithelium (for example, the agent can be in a skin patch or other apparatus, and the vehicle may be contained in a liquid composition or other type of composition that is applied to the skin prior to application of the skin patch). As used in the present invention, the word "therapeutic" within the context of blood glucose, refers to a decrease in blood glucose levels sufficient to alleviate symptoms or acute signs of hyperglycemia, for example, in diabetic patients. In certain aspects of the present invention, the transdermal delivery of therapeutic proteins with the ability to achieve therapeutic alterations of blood glucose is specifically excluded. The present invention specifically excludes antibody fragments that have no biological activity in addition to only binding a specific antigen, when the term "therapeutic" or "biologically active protein" is used. Since the antigens suitable for immunization have other biological activities such as assembly of an immune response, these remain included in the appropriate aspects of the present invention. In addition, agents that have a biological activity or a therapeutic effect by binding a specific antigen, or thereby blocking the ligand linkage or altering the conformation of the antigen, are included in the present invention. As used in the present invention, antigenic agents suitable for immunization can be protein-based antigens that do not alter in blood glucose levels, agents without nucleic acid, without protein or hybrids thereof. The nucleic acids encoding antigens are specifically unsuitable for the compositions of the present invention. Therefore, the included agents are the proper antigens for immunization. Suitable antigens include, for example, those for environmental, pathogenic or biologically hazardous agents. The agents suitable include, for example, antigens related to botulism, malaria, rabies, anthrax, tuberculosis or related to childhood immunizations such as hepatitis B, diphtheria, pertussis, tetanus, Haemophilus influenza type b, inactivated poliovirus, measles, mumps, rubella, chicken pox, pneumococcus, hepatitis A and influenza. Skeleton positively charged The positively charged skeleton (also referred to as a positively charged "vehicle") is usually a linear chain of atoms, either with groups in the chain carrying a positive charge at physiological pH, or with groups that carry a positive charge attached to the side chains that extend from the skeleton. Preferably, the skeleton charged in a positive manner by itself will not have a defined enzymatic or biological activity. The linear skeleton is a hydrocarbon skeleton which, in some modalities, it is interrupted by heteroatoms selected from nitrogen, oxygen, sulfur, silicon and phosphorus. Most skeletal chain atoms are usually carbon. In addition, the backbone will often be a polymer of repeat units (e.g., amino acids, poly (ethyleneoxy), poly (propyleneamine), polyalkylenenoamine, and the like). In a group of embodiments, the positively charged skeleton is a polypropyleneamine wherein a number of amine nitrogen atoms are found as ammonium (tetra-substituted) groups bearing a positive charge. In another modality, the loaded skeleton in positive form it is a peptidyl-free polymer, which may be a hetero or homopolymer, such as a polyalkyleneimine, for example, a polyethyleneimine or polyalkyleneimine, having a molecular weight of from about 10,000 to about 2,500,000, preferably from about 100,000 to about 1 , 800,000, and most preferably from about 500,000 to about 1,400,000. In another group of embodiments, the backbone has attached a plurality of side chain portions including positively charged groups (eg, ammonium groups, pyridino groups, phosphonium groups, sulfonium groups, guanidinium groups, or amidinium groups). The side chain portions in this group of modalities can be placed in spacings along the skeleton, which are consistent in the separations or variables. In addition, the length of the side chains can be similar or dissimilar. For example, in a group of embodiments, the side chains may be straight or branched hydrocarbon chains having from 1 to 20 carbon atoms and ending in the distal end (away from the backbone) in one of the positively charged groups. aforementioned. In all aspects of the present invention, the association between the carrier and the biologically active agent is by non-covalent interaction, which may include, for example, ionic interactions, hydrogen bonding, van der Waals forces or combinations thereof .

In one group of embodiments, the positively charged backbone is a polypeptide having multiple side chain groups charged in positive form (eg, lysine, arginine, ornithine, homoarginine, and the like). Preferably, the polypeptide has a molecular weight of from about 10,000 to about 1,500,000, more preferably from 25,000 to about 1,200,000, most preferably from about 100,000 to about 1,000,000. One skilled in the art will appreciate that when amino acids are used in this part of the present invention, the side chains can have either D or L form (R or S configuration) at the center of the adhesion. Alternatively, the backbone can be an analogue of a polypeptide, such as a peptoid. See for example the publication of Kessier, Angew. Chem. Int. Ed. Engl. 32: 543 (1993); Zuckermann and Associates, Chemtracts-Macromol. Chem. 4:80 (1992); and Simón y Asociados, Proc. Natl. Acad. Sci. USA 89: 9367 (1992)). In synthesis, a peptoide is a polyglycine in which the side chain is attached to the nitrogen atoms of the skeleton instead of the carbon-a atoms. As described above, a portion of the side chains will usually end up in a positively charged group to provide a positively charged backbone component. Peptoid synthesis is described, for example, in U.S. Patent No. 5,877,278. As used in the present invention, the term skeletons positively charged that have a peptoid skeleton construction, are considered "without peptides", since they are not composed of amino acids that have side chains that naturally occur at the α-carbon locations. A variety of other backbones can be used, for example, spherical or electronic polypeptide mimetics, wherein the amide ligatures of the peptide are replaced with substitutes such as ester ligatures, thioamides (-CSNH-), reverse thioamides (-NHCS- ), aminomethylene (-NHCH2-), or the reverse methylene-amino groups (-CH2NH-), keto-methylene groups (-COCH2-), phosphinate (-PO2RCH2-), phosphonoamidate or phosphonamidate ester (-PO2RNH-), reverse peptide (-NHCO-), trans-alkene (-CR = CH-), fluoroalkene (-CF = CH-), dimethylene (-CH2CH2-), thioether (-CH2S-), hydroxyethylene (-CH (OH) CH2-) , methyleneoxy (-CH2O) -, tetrazole (-CN4), sulfonamido (-SO2NH-), methylenesulfonamido (-CHRSO2NH-), reverse sulfonamide (-NHSO2-), and skeletons with malonate and / or gem-diamino-alkyl subunits, for example, as indicated in the publication of Fletcher and Associates ((1998) Chem. Rev. 98: 763) and as detailed through the references mentioned here. Many of the above substitutions result in approximately isosteric polymer skeletons relative to the backbones formed from amino acids-a. In each of the skeletons provided above, the side chain groups can be attached since they carry a positively charged group. For example, skeletons linked with sulfonamide (-SO2NH- and -NHSO2-) may have side chain groups attached to the nitrogen atoms. Similarly, the hydroxyethylene ligation (-CH (OH) CH2-) may contain a side chain group attached to the hydroxy substituent. One skilled in the art can easily adapt the other ligation chemistries to provide side chain groups positively charged using standard synthetic methods. In a particularly preferred embodiment, the positively charged skeleton is a polypeptide having branching groups (also referred to as efficiency groups) independently selected from - (gly) n? - (arg) n2, HIV TAT or fragments thereof , or the AntenNapedia protein transduction domain or a fragment or mixture thereof, wherein the subscript n1 is an integer from 0 to 20, more preferably from 0 to 8, even more preferably from 2 to 5, and the subscript n2 is independently an integer non of from about 5 to about 25, more preferably from about 7 to about 17, most preferably from about 7 to about 13. Preferred still further, are the modalities in which the HIV-TAT fragment has the formula (gly) P-RGRDDRRQRRR- (gly) q, (gly) p-YGRKKRRQRRR- (gly) q, or (gly) p-RKKRRQRRR- (gly) q, wherein the subscripts p and q are each independently an integer from 0 to 20 and the fragment adheres to the skeleton through either the C-terminus or the N-terminus of the fragment. Preferred HIV-TAT fragments are those in which the subscripts p and q are each independently integers from 0 to 8, more preferably from 2 to 5. In another preferred embodiment, the side chain charged in positive form or the branching group is the protein transduction domain Antennapedia (Antp) (PTD) or a fragment thereof that retains the activity. Preferably, the positively charged vehicle includes branching groups charged in positive side chain form in an amount of at least about 0.05%, such as a percentage of the total vehicle weight, preferably from about 0.05 to about 45% by weight and most preferably from about 0.1 to about 30% by weight. For positively charged branching groups having the formula - (gly) n? - (arg) n2, the most preferred amount is from about 0.1 to about 2.5%. In another particularly preferred embodiment, the backbone part is a polylysine and the positively charged branching groups adhere to the lysine side chain amino groups. The polylysine used in this particularly preferred embodiment has a molecular weight of from about 10,000 to about 1,500,000, preferably from about 25,000 to about 1,200,000, and most preferably from 100,000 to about 1,000,000. It can be any of the polylysines commercially available (Sigma Chemical Company, St. Louis, Missouri, USA) such as, for example, polylysine having MW > 70,000, polylysine having MW of 70,000 to 150,000, polylysine having MW 150,000 to 300,000, and polylysine having MW > 300,000 The proper selection of a polylysine will depend on the remaining components of the composition, and will be sufficient to provide a total net positive charge to the composition and provide a length that is preferably from 1 to 4 times the combined length of the negatively charged components. . Branched groups loaded in positive form or preferred efficiency groups include, for example, -gly-gly-gly-arg-arg-arg-arg-arg-arg-arg (-Gly3Arg7) or HIV-TAT. In another preferred embodiment, the positively charged backbone is a long chain polyalkyleneimine such as a polyethyleneimine, for example one having a molecular weight of about 1., 000,000. Skeletons or positively charged carrier molecules comprising polymers of polypeptides or polymers without peptidyl, such as polyalkylene imines and other positively charged backbones mentioned above, having branching gr described above, are novel compounds and form an aspect of the present invention. invention. In one embodiment of the present invention, only a positively charged vehicle having positively charged branching gr is necessary for the transgenic delivery of the active substance. In a modality of this case, the positively charged vehicle is a polypeptide (eg, lysine, arginine, ornithine, homoarginine and the like) having side chain groups charged in multiple positive form, as described above. Preferably, the polypeptide has a molecular weight of at least 10,000. In another embodiment of this case, the positively charged vehicle is a peptidyl-free polymer such as polyalkyleneimine having multiple side-chain groups positively charged having a molecular weight of at least about 100,000. Said polyalkyleneimines include polyethylene and polypropyleneimines. In any case, to be used as the only agent necessary for transdermal delivery, the positively charged transported molecule includes branching or efficiency groups loaded in positive form, comprising - (gly) n? - (arg) p2, wherein the subscript n1 is an integer from 0 to 20, more preferably from 0 to 8, still more preferably from 2 to 5, and the subscript n2 is independently an integer non from about 5 to about 25, more preferably from about 7 to about 17 , and most preferably from about 7 to about 13, HIV-TAT or fragments thereof, or an Antennapedia PTD or fragment thereof. Preferably, the side chain or branching groups have the formula - (gly) n? - (arg) n2, tai as described above. Other preferred embodiments are those in which the branching or deficiency groups are fragments (gly) p-RGRDDRRQRRR- (gly), (gly) p-YGRKKRRQRRR- (gly) q, or (gly) p-RKKRRQRRR- (gly) q, wherein the subscripts p and q are each independently an integer from 0 to 20, and the fragment is adhered to the transporter molecule through either the C-terminus or the N-terminus of the fragment. The lateral branching groups may have either D- or L-shape (R or S configuration) at the center of the adhesion. The preferred HIV-TAT fragments are those in which the subscripts p and q are each independently integers from 0 to 8, more preferably from 2 to 5. Other preferred embodiments are those wherein the branching groups are Antennapedia PTD groups or fragments of the they retain the activity of the group. These are known in the art, for example, from the publication of Consolé y Asociados, J. Biol. Chem. 278: 35109 (2003). In a particularly preferred embodiment, the carrier is a polylysine with branching groups charged in a positive manner adhered to the amino groups of the lysine side chain. The polylysine used in this particularly preferred embodiment can be any of the commercially available polylysines (Sigma Chemical Company, St. Louis, Missouri, USA) such as for example polylysine having MW > 70,000, polylysine having MW of 70,000 to 150,000, polylysine having MW 150,000 to 300,000 and polylysine having MW > 300,000 However, preferably polylysine has MW of about 10,000. Branching groups or efficiency groups loaded in positive form include for example -gly-gly-gly-arg-arg-arg-arg-arg-arg-arg (-Gly3Arg7) or H IV-TAT or fragments thereof and Antennapedia PTD or fragments thereof. Other components In addition to the positively charged backbone component, the multiple component compositions of the present invention comprise at least two components of the group consisting of the following: i) a first backbone negatively charged having a plurality of backbone portions; generation of adhered images; or alternatively a plurality of negatively charged image generation portions; ii) a second negatively charged skeleton having a plurality of adhered steering agents; or alternatively, a plurality of negatively charged address portions; iii) at least one selected member of RNA, DNA, ribozymes, modified oligonucleic acids and cDNA encoding a selected transgene; iv) DNA encoding at least one persistence factor; and v) a third skeleton loaded in a negative form having a plurality of biological agents adhered, or a biological agent loaded in a negative form. In a related aspect, as defined in the present invention, in some embodiments or compositions herein invention, the skeleton or positively charged vehicle can be used only to provide the transdermal delivery of certain types of substances. Combinations of biologically active agents as described in the present invention, such as, for example, combinations of insulin, botulinum toxin, proteins that do not alter blood glucose levels in a therapeutic manner, antigens suitable for immunization or agents without acid nucleic acids, without protein, can also be used in these compositions. Negatively charged skeletons, when used to carry the image generating portions, targeting portions and therapeutic agents, can be a variety of skeletons (similar to those described above) having multiple groups carrying a negative charge in a physiological pH Alternatively, the image generation portions, therapeutic agent targeting portions with sufficient negatively charged portions on the surface, will not require addition of an additional skeleton for ion complex with positively charged skeletons, as you will readily appreciate. those skilled in the art. The term "sufficient" within this context implies that an adequate density of negatively charged groups is found on the surface of the image generating portions, targeting portions or therapeutic agents to produce an ionic bond with the skeletons charged in the form Positive described above. In these cases, the substance derived therefrom, it has sufficient negative charge to associate with the positively charged carriers of the present invention in non-covalent form. The term "sufficient" within this context can be determined, for example, through a change in a particle size or functional spectrophotometry versus the components alone. Suitable negatively charged groups are carboxylic acids, phosphinic, phosphonic or phosphoric acids, sulphonic or sulphonic acids and the like. In 'some embodiments, the skeleton loaded in negative form will be an oligonucleotide. In other embodiments, the skeleton charged in a negative form is an oligosaccharide (e.g., dextran). In still other embodiments, the skeleton charged in negative form is a polypeptide (eg, poly-glutamic acid, polyaspartic acid or polypeptide wherein the glutamic acid or aspartic acid residues are disrupted through uncharged amino acids). The portions described in more detail below (image generation portions, targeting agents and therapeutic agents) can adhere to a skeleton having these pendant groups, usually through ligatures thereof. Alternatively, the amino acids that interrupt the negatively charged amino acids or that adhere to the negatively charged skeleton terminus, can be used to adhere image generation portions and address portions, for example, by ligatures of disulfide (through a cysteine residue), amide ligatures, or ligatures (through hydroxyl groups of serine or threonine and the like). Alternatively, the image generation portions and address portions, by themselves may be small anions in the absence of a negatively charged polymer. Alternatively, the image generation portions, targeting portions and therapeutic agents may themselves be covalently modified to produce sufficient negatively charged portions at the surface for the ionic complex with the positively charged skeletons, such as it will be appreciated by those skilled in the art. In both of these cases, the substance or a derivative thereof have sufficient negative charge to associate with the positively charged carriers of the present invention in non-covalent form. The term "sufficient" within this context, refers to an association that can be determined, for example, by change in particle size or functional spectrophotometry versus the components alone. Portions of image generation In the present invention, a variety of diagnostic or imaging portions are useful, and are found in an effective amount that will depend on the conditions being diagnosed or on which the image is being generated, the route of administration, the sensitivity of the agent or apparatus used for the detection of the agent, and the like. Examples of suitable imaging or diagnostic agents include radiopaque contrast agents, paramagnetic contrast agents, superparamagnetic contrast agents, optical image generation portions, CT contrast agents and other contrast agents. For example, radiopaque contrast agents (for X-ray image generation, will include organic and inorganic iodine compounds (e.g., diatrizoate), radiopaque metals and their salts (e.g., silver, gold, and platinum, and the like) and other radiopaque compounds (e.g., calcium salts, barium salts, such as barium sulfate, tantalum, and tantalum oxide.) Suitable paramagnetic contrast agents (for MR imaging) include gadolinium diethylene triaminopentaacetic acid ( Gd-DTPA) and its derivatives, and other complexes of gadolinium, manganese, iron, dysprosium, copper, europium, erbium, chromium, nickel and cobalt, including complexes with 1, 4,7, 10-tetra-azacyclododecane-N, N ', N ", N'" - tetra-acetic (DOTA), ethylenediaminetetraacetic acid (EDTA), 1, 4,7, 10-tetra-azacyclododecane-N, N ', N ", N'" - triacetic (DO3A), 1, 4,7-triazacyclononane-N, N ', N "-triacetic (NOTE), 1, 4,8, 1 1 -tetra-azacyclotetradecane-N, N', N", N '"- tetraacetic acid (TETA) ), diacetic acid hydroxybenzylethylenediamine (HBED) and the like Suitable superparamagnetic contrast agents (for MR imaging) include magnetites, superparamagnetic iron oxides, monocrystalline iron oxides, particularly compound forms of each of these agents that can adhere to a negatively charged skeleton, yet other suitable imaging agents are the CT contrast agents including iodinated and non-iodinated and ionic and nonionic CT contrast agents, as well as contrast agents such as spin labels or other diagnostically effective agents. Suitable optical imaging agents include for example the group consisting of Cy3, Cy3.5, Cy5, Cy5.5, Cy7, Cy7.5, Oregon green 488, Oregon green 500, Oregon, green 514, green fluorescent protein , 6-FAM, Red Texas, Hex, TET, and HAMRA. Other examples of diagnostic agents include marker genes that encode proteins that are readily detectable when expressed in a cell, including but not limited to beta-galactosidase, green fluorescent protein, blue fluorescent protein, luciferase, and the like. A wide variety of labels can be employed, such as radionuclides, fluoros, enzymes, enzyme substrates, enzyme cofactors, enzyme inhibitors, ligands (particularly haptens), and the like. Still other useful substances are those labeled in radioactive species or components, such as "mTc glucoheptonate." The choice of adhering an image generation portion to a negatively charged skeleton will depend on a variety of conditions. neutral at a physiological pH and will preferably adhere to a skeleton loaded in a negative form or covalently modified to include enough negatively charged portions of the above, to retain a complex with the vehicle Loaded in a positive way Other imaging agents carry sufficient negative charge to retain the complex with the positively charged vehicle, even in the absence of a negatively charged skeleton. In these cases, the substances or derivatives thereof have sufficient negative charge to associate with the positively charged vehicles of the present invention in non-covalent form. The term "sufficient", within this context, refers to an association that can be determined, for example, through the change in particle size or functional spectrophotometry versus the components alone. Examples of said negatively charged image generation portions include phosphate ion (useful for magnetic resonance imaging). Steering Agents A variety of steering agents are useful in the compositions described herein. Usually, the steering agents re to a negatively charged skeleton as described for the above image generation portions. In certain embodiments, the steering agents and the portions of image generation are structurally and / or chemically distinct. For example, the image generation portions and steering agents are not both phosphate. Generally, the targeting agents can be any element that makes it possible to direct the transfer of a nucleic acid, therapeutic agent or other component of the composition to a particular site or to alter the tropism of the complex in relation to that of the complex without the agent of direction. The targeting agent can be an extracellular targeting agent, which allows, for example, directing a transfer of nucleic acid to certain types of cells or certain desired tissues (tumor cells, liver cells, hepatopoietic cells and the like) . Said agent can also be an intracellular targeting agent, which allows a therapeutic agent to be targeted to particular cellular compartments (eg, mitochondria, nucleus and the like). The simplest agent must also be a small anion, which, by changing the net charge distribution, alters the tropism of the complex from higher-level negative cell surfaces and extracellular matrix components, to a wide variety of cells or even specifically out of the higher level negative surfaces. The targeting agent or agents are preferably linked, covalently or non-covalently, to a skeleton charged in a negative form in accordance with the present invention. According to a preferred mode of the present invention, the targeting agent is covalently adhered to a sulfated or phosphorylated nucleic acid, polyaspartate, or dextran and the like which serves as a negatively charged backbone component, preferably through a liaison group. Methods for adhering targeting agents (as well as other biological agents) to nucleic acids are known to those skilled in the art, using, for example, heterobifunctional linking groups (see publication of Pierce Chemical Catalog.). In a group of embodiments, the targeting agent is a fusogenic peptide to promote cellular transfection, that is, to favor the passage of the composition or its various elements through the membranes, or to assist in the exit of endosomes or to cross over the nuclear membrane. The targeting agent may also be a cellular receptor ligand for a receptor that is on the surface of the cell type, such as, for example, a sugar, transferrin, insulin or asialo-orosomucoid protein. Said ligand may also be one of intracellular type, such as nuclear location signal sequence (nls) which promotes the accumulation of transfected DNA within the nucleus. Other useful agents within the context of the present invention include sugars, peptides, hormones, vitamins, cytosines, nucleic acids, small anions, lipids, or sequences or fractions derived from these elements and which allow specific binding to their corresponding receptors. Preferably, the targeting agent is sugars and / or peptides such as antibodies or antibody fragments, cell receptor ligands, or fragments of the same receptors or fragments of receptors and the like. More preferably, the targeting agents are ligands of growth factor receptors, cytosine receptors, or cell lecithin receptors or adhesion protein receptors. The steering agent can also be a sugar which makes it possible to direct lecithins such as asialo-glycoprotein receptors, or alternatively an antibody or Fab fragment which makes it possible to target the immunoglobulin Fe fragment receptor. In still other embodiments, a steering agent is used in the absence of a negatively charged skeleton. Within this group of embodiments, the steering agent carries enough negatively charged portions to retain an ionic complex with the positively charged vehicle described above. In these cases, the substance or derivative thereof has sufficient negative charge to associate with the positively charged carriers of the present invention in non-covalent form. The term "sufficient" within this context refers to an association that can be determined, for example through the change in particle sizes or functional spectrophotometry versus the components alone. Negatively charged targeting agents for this group of embodiments are protein-based targeting agents that have a net negative charge at a physiological pH, as well as targeting agents that can facilitate adhesion to a particular cell surface. , such as small polyanions including for example phosphate, aspartate and citrate which can, for example, change the direction based on the charge of the net surface area of the cell to be targeted. In the composition of the present invention, the nucleic acid can be either a deoxyribonucleic acid or an acid ribonucleic acid, and may comprise sequences of natural or artificial origin. More particularly, the nucleic acids used in the present invention may include DNA, cDNA, mRNA, tRNA, genomic rRNA, hybrid or synthetic sequences or semi-synthetic sequences. These nucleic acids may be of human, animal, plant, bacterial, viral, etc. origin. In addition, the nucleic acids can be obtained by any technique known to those skilled in the art, and in particular by bank sorting, by chemical synthesis or by mixing methods that include chemical or enzymatic modification of sequences obtained by classifying the banks. Still further, nucleic acids can be incorporated into vectors, such as plasmid vectors. The deoxyribonucleic acids used in the present invention may be single-stranded or double-stranded. These deoxyribonucleic acids can also encode therapeutic genes, sequences for regulating transfection or replication, antisense sequences, regions for binding to other cellular components, etc. Suitable therapeutic genes are essentially any gene that encodes a protein product that has a therapeutic effect. The protein product encoded in this way can be a protein, polypeptide, peptide or the like. The protein product may, in some cases, be homologous with respect to the targeting cell (i.e. a product that is normally expressed in the target cell when the latter does not exhibit pathology). In this way, the use of suitable nucleic acids can increase the expression of a protein, making it possible, for example, to overcome insufficient expression in the cell. Alternatively, the present invention provides compositions and methods for the expression of a protein that is inactive or weakly active due to a modification, or alternatively to overexpress the protein. The therapeutic gene can thus encode a mutant of a cell protein, which has increased stability, modified activity, etc. The protein product can also be heterologous with respect to the target cell. In this case, an expressed protein can, for example, elaborate or provide an activity which is deficient in the cell, allowing to combat a pathology or to stimulate an immune response. More particularly, the nucleic acids useful in the present invention are those encoding enzymes, blood derivatives, hormones, lymphokines, interleukins, interferons, TNFs, growth factors, neurotransmitters or their precursors or synthetic enzymes, or trophic factors: BDNF, CNTF, NGF, IGF, GMF, aFGF, bFGF, VEGF, NT3, NT5, HARP / pleiotropin; proteins involved in lipid metabolism, apolipoprotein type selected from apolipoproteins Al, Al I, A-IV, B, Cl, Cl l, Cl ll, D, E, F, G, H, J, and apo ( a), metabolic enzymes such as, for example, lipoprotein lipase, hepatic lipase, lecithin cholesterol acyltransferase, 7-a-cholesterol hydroxylase, phosphatidic acid phosphatase or lipid transfer proteins such as cholesterol ester transfer proteins and phospholipid transfer proteins, a protein for binding HDLs or a receptor selected from, for example, LDL receptors, receptors with chylomicron residues and purifying receptors, dystrophin or minidistrofin, GAX protein, associated CFTR protein with mucoviscidosis, tumor suppressor genes; p53, Rb, Rapl A, DCC, k-rev; protein factors involved in coagulation: factors VII, VI II, IX; or the nucleic acids may be the genes involved in DNA repair, suicide genes (thymidine kinase, cytosine diaminase), genes encoding thrombomodulin, α1-antitrypsin, tissue plasminogen activator, superoxide dismutase, elastase, metalloproteinase matrix and the like. Therapeutic genes useful in the present invention, they can also be an antisense sequence or a gene whose expression in the target cell makes it possible to control the expression of genes or the transcription of the cellular mRNA. For example, said sequences can be transcribed in the target cell into RNA complementary to the cellular mRNA, and thus block its translation into protein, according to the technique described in EP 140,308. The antisense sequences also comprise the sequences encoding ribozymes, which have the ability to selectively destroy the target RNA (see EP 321, 201). As indicated above, the biologically active agent may also comprise one or more antigenic peptides that have the ability to generate an immune response in humans or animals. In this particular embodiment, the present invention makes it possible in this way to produce either vaccines or immunotherapeutic treatments to humans or animals, in particular against microorganisms, viruses, or cancers. They can be in particular antigenic peptides specific for Epstein-Barr virus, for virus VI H, for hepatitis B virus (see EP 185,573), for pseudo-rabies virus or as an alternative specific for tumors (see Patent EP 259,212) . Preferably, the nucleic acid also comprises sequences that allow the expression of the therapeutic gene and / or the gene encoding the antigenic peptide in the desired cell or organ. These may be sequences that are naturally responsible for the expression of the gene considered when these sequences have the ability to function in the infected cell. The nucleic acids can also be sequences of different origin (responsible for the expression of other proteins, or even synthetic proteins). In particular, the nucleic acids may contain promoter sequences of eukaryotic or viral genes. For example, the promoter sequences may be those derived from the genome of the cell to be infected. Similarly, the promoter sequences can be derived from the genome of a virus, for example, the promoters of the EIA, MLP, CMV, RSV, etc. genes. In addition, these expression sequences can be modified through the addition of activation sequences, sequences of regulation, etc. In addition, the nucleic acid may also contain, in particular the upstream of the therapeutic gene, a signal sequence that directs the synthesized therapeutic product within the secretion trajectories of the target cell. This signal sequence may be the natural signal sequence of the therapeutic product, although it may also be any other functional signal sequence, or an artificial signal sequence. DNA encoding at least one persistence factor In some embodiments, the composition will also comprise DNA that codes for at least one persistence factor. The example of said DNA is the DNA encoding the pre-terminal adenoviral protein 1 (see the publication by Lieber, et al., Nature Biotechnology 15 (13): 1383-1387 (1997).) The pre-terminal adenoviral protein 1 or the nucleic acid it encodes it, can be provided in cis- or trans- to the nucleic acid sequence encoding the desired therapeutic transgene.When provided in this manner, the pre-terminal protein 1 or the sequence retains the therapeutic nucleic acid in the form of an episome This means that a variety of biological agents, including both therapeutic and cosmetological agents, are useful in the present invention and prevent further decreases in the expression of therapeutic protein. effective amount that it will depend on the condition that will be treated, prophylactically or otherwise, the route of administration, the effectiveness of the agent and the constitution of the patient and the susceptibility to the treatment regimen. Suitable therapeutic agents that can be adhered to a negatively charged skeleton can be found in essentially any class of agents, including, for example, analgesic agents, anti-asthmatic agents, antibiotics, antidepressant agents, anti-diabetic agents, antifungal agents, antiemetics, antihypertensives, anti-potency agents, anti-inflammatory agents, antineoplastic agents, anti-HIV agents, antiviral agents, anxiolytic agents, contraceptive agents, fertility agents, antithrombotic agents, prothrombotic agents, hormones, vaccines, immunosuppressive agents, vitamins and similar. Alternatively, sufficient negatively charged groups can be introduced into the therapeutic agent to produce the ionic complex with the positively charged backbones described above. There are many suitable methods, such as phosphorylation and sulfation, as can be appreciated by those skilled in the art. In addition, certain agents themselves possess adequate portions negatively charged to associate with the positively charged vehicle described above, and do not require adhesion to a negatively charged backbone. In these cases, the substance or a derivative thereof has sufficient negative charge to associate with the positively charged vehicles of the present invention. The term "sufficient" within this context refers to an association that can be determined, for example, through the change in particle size or functional spectrophotometry versus the components alone. Suitable cosmetogenic agents include, for example, epidermal growth factor (EGF), as well as human growth hormone, antioxidants and botulinum toxin. Within the context of the present invention, the term "botulinum toxin" includes not only the botulinum pseudotypes A, B, C, D, E, F and G, but also fragments thereof having botulinum light chain activity. More particularly, the therapeutic agents useful in the present invention include analgesics such as lidocaine, novocaine, bupicaine, procaine, tetracaine, benzocaine, cocaine, mepivacaine, etidocaine, proparacaine, ropivacaine, prilocaine and the like; anti-asthmatic agents such as azelastine, cetotifen traxanox, corticosteroids, cromolyn, nedocromil, albuterol, bitolterol mesylate, pirbuterol, salmeterol, terbutilin, theophylline and the like; antibiotic agents such as neomycin, streptomycin, chloramphenicol, norfloxacin, ciprofloxacin, trimethoprim, sulfamethyloxazole, ß-lactam antibiotics, tetracycline and the like; antidepressant agents such as nefopam, oxypertine, imipramine, trazadone and the like; anti-diabetic agents, such as biguanidines, sulfonylureas and the like; antiemetics and antipsychotics such as chlorpromazine, flufenacin, perphenazine, prochlorperazine, promethazine, tiethylperazine, triflupromazine, haloperidol, scopolamine, diphenidol, trimethobenzamide and the like; neuromuscular agents such as atracurium mivacurium, rocuronium, succinylcholine, doxacurium, tubocurarine and botulinum toxin (BTX); antifungal agents such as amphotericin B, nystatin, candicidin, itraconazole, ketoconazole, miconazole, clotrimazole, fluconazole, cyclopirox, econazole, naftifine, terbinafine, griseofulvin, ciciopirox and the like; antihypertensive agents such as propanolol, propafenone, oxyprenolol, nifedipine, reserpine and the like; anti-impotence agents such as nitric oxide donors and the like; anti-inflammatory agents including spheroidal anti-inflammatory agents such as cortisone, hydrocortisone, dexamethasone, prednisolone, prednisone, fluazacort and the like, as well as non-steroidal anti-inflammatory agents such as idometacin, ibuprofen, ramidenizone, prioxicam and the like; antineoplastic agents such as adriamycin, cyclophosphamide, actinomycin, bleomycin, duanoribicin, doxorubicin, epirubicin, mitomycin, rapamycin, methotrexate, fluorouracil, carboplatin, carmustine (BCNU), cisplatin, etoposide, interferons, phenesterin, taxol, (including analogues and derivatives), camptothecin and derivatives thereof, vinblastine, vincristine and the like; anti-VI H agents (eg, antiproteolytics); antiviral agents such as amantadine, metisazone, idoxuridine, cytarabine, acyclovir, famciclovir, ganciclovir, foscarnet, sorivudine, trifluridine, valaciclovir, cidofovir, didanosine, stavudine, zalcitabine, zidovudine, ribavirin, rimantatin and the like; anxiolytic agents such as dantrolene, diazepam, and the like; COX-2 inhibitors; contraceptive agents such as progestogen and the like; anti-thrombotic agents such as GPIIb / l, inhibitors, tissue plasminogen activators, streptokinase, urokinase, heparin and the like; prothrombotic agents such as thrombin, factors V, Vi l, VI II and the like; hormones such as insulin, growth hormone, prolactin, EGF (epidermal growth factor) and the like; immunosuppressive agents such as cyclosporin, azathioprine, mizorobin, FK506, prednisone and the like; angiogenic agents such as VEGF (vascular endothelial growth factor); vitamins such as A, D, E, K and the like; and other therapeutically or medicinally active agents. See for example the publication of GOODMAN & GILMAN'S THE PHARMACOLOGICAL BASIC OF THERAPEUTICS, Ninth Edition, Hardman, et al., Eds. McGraw-Hill, (1996). In the most preferred embodiments, the biological agent is selected from insulin, botulinum toxin, VEGF, antigens for immunization and antifungal agents. As noted above for the targeting agents and imaging agents, certain biological or cosmetological agents may be used in the absence of a negatively charged skeleton. These biological agents or Cosmetological are those that generally carry a net negative charge at the physiological pH to retain the complex with the positively charged vehicle. Examples include botulinum toxin (a large MW protein), insulin (a small MW protein), immunization antigens, which can range from very small to very large and typically include proteins or glycoproteins, and many antifungal agents. In these cases, the substance or derivative thereof has a sufficient negative charge to associate with the positively charged carriers of the present invention in non-covalent form. The term "sufficient" within this context refers to an association that can be determined, for example, by the change in particle size or functional spectrophotometry versus the components alone. Negatively charged skeletons having imaging generation portions, targeting agents and adherent therapeutic agents For three of the above component groups, including image generation portions, targeting agents and therapeutic agents, individual compounds can be adhered to a skeleton charged in a negative form, modified covalently to introduce negatively charged portions, or used directly if the compound contains sufficient negatively charged portions to confer ionic bond to the positively charged backbone described above. When is necessary, normally the adhesion is through a linking group used to covalently adhere the particular agent of the structure through functional groups present in the agent, as well as the backbone. A variety of linking groups are useful in this aspect of the present invention. See, for example, the publications Hermanson, Bioconjugate Techniques, Academic Press, San Diego, CA (1996); Wong, S.S. , Ed., Chemistry of Protein Conjugation and Cross-Linking, CRC Press, Inc., Boca Raton, FL (1991); Senter, et al., J. Org. Chem. 55: 2975-78 (1990); and Koneko, et al., Bioconjugate Chem. 2: 133-141 (1991). In some embodiments, the therapeutic, diagnostic or targeting agents will not have a functional group available to adhere to a linking group, and may be modified first to incorporate, for example, a hydroxy, amino or thiol substituent. Preferably, the substituent is provided in a portion without interference from the agent, and can be used to adhere a linking group, and will not adversely affect the function of the agent. In yet another aspect, the present invention provides compositions comprising a non-covalent complex in a positively charged backbone having at least one adhered efficiency group and at least one nucleic acid member selected from the group consisting of RNA, DNA, ribozymes, modified oligonucleic acids and cDNA encoding a selected transgene. In this aspect of the present invention, the skeleton positively charged can essentially be any of the positively charged backbones described above, and will also comprise (as with the previously selected backbones) at least one adhered efficiency group. Suitable efficiency groups include, for example, (Gly) n? - (Arg) n2, wherein the subscript n 1 is an integer from 3 to about 5, and the subscript n2 is independently an integer non of from about 7 up to approximately 17 or TAT domains. In addition, the nucleic acids useful in this aspect of the present invention are the same as described above. Transdermal supply of insulin and certain larger molecules It has been found that previously positively charged vehicles can be used for transdermal delivery of insulin and certain other biologically active agents that do not alter blood glucose levels in a therapeutic manner., such as proteins having a molecular weight of about 50,000 and higher, for example, botulinum toxin (BTX), or for other biologically active agents such as therapeutic nucleic acid-based agent, a therapeutic agent without nucleic acid, without such protein as certain antifungal agents, or alternatively, an agent for immunization. The use of the positively charged vehicle allows the transmission of the marker protein or gene both inside and outside skin cells, and the delivery thereof in an effective amount and active form for an underlying tissue. For example, insulin can be delivered through the skin into the underlying capillaries for transport through the body without the need for injection. Botulinum toxin can be delivered to underlying muscles or glandular structures within the skin in an amount effective to produce paralysis, produce relaxation, relieve contractions, prevent or relieve spasms, reduce glandular production or provide other desired effects. Local delivery in this form can produce reductions in dosage, reduce toxicity, and allow more precise dose optimization for the desired effects in relation to injectable or implantable materials, particularly in the case of botulinum toxin. This embodiment may include an amount of a preferably small polyvalent anion, for example, phosphate, aspartate or citrate, or it may be carried out in the substantial absence of said polyanion. In all aspects of the present invention, the association between the carrier and the biologically active agent is by non-covalent interaction, which may include, for example, ionic interactions, hydrogen bonding, van der Waals forces, or combinations of the same. The term "botulinum toxin" as used in the present invention, refers to any of the known serotypes of botulinum toxin, either produced through the bacterium or by recombinant techniques, as well as any types that may be subsequently discovered including variants or fusion proteins constructed. As mentioned previously, at this time, seven immunologically distinct botulinum neurotoxins have been characterized, ie serotypes of botulinum neurotoxin A, B, C, D, E, F and G, each of which is distinguished by neutralization with specific antibodies to the type. Botulinum toxin serotypes are available from Sigma-Aldrich and from Metabiologics, Inc., (Madison, Wisconsin), as well as from other sources. The different serotypes of botulinum toxin vary in the animal species that affect them, and in the severity and duration of the paralysis they cause. At least two types of botulinum toxin, types A and B, are usually commercially available in formulations for treatments of certain conditions. Type A, for example, is contained in preparations of Allergan that have the trade name BOTOX®, and of Ipsen that have the trade name DYSPORT®, and type B is contained in preparations of Elan that has the trade name MYOBLOC®. The botulinum toxin used in the compositions of the present invention can be a botulinum toxin derivative, which is a compound that has botulinum toxin activity but contains one or more chemical or functional alterations in any part or in any chain in relation to botulinum toxins that occur naturally with native recombinants. For example, botulinum toxin can be a modified neurotoxin, which is a neurotoxin that has at least one of its amino acids removed, modified or replaced, compared to a native neurotoxin, or the modified neurotoxin may be a neurotoxin produced recombinantly or a derivative or fragment thereof. For example, the botulinum toxin may be one that has been modified in such a way that, for example, it increases its properties or decreases the undesirable side effects, but still retains the desired activity of the botuiinic toxin. The botulinum toxin can be any of the botulinum toxin complexes produced through the bacterium, as described above. Alternatively, the botulinum toxin can be a toxin prepared using recombinant or synthetic chemical techniques, for example, a recombinant peptide, a fusion protein or a hybrid neurotoxin, for example, prepared from subunits or domains of different pseudotypes of botulinum toxin. (See US Patent No. 6, 444,209, for example). The botulinum toxin may be a part of the general molecule that has been shown to possess the necessary activity of botulinum toxin, and in such case it may be used as such or as part of a combination or complex molecule, for example, a fusion protein. Alternatively, a portion of the toxin can be used directly with the positively charged backbones described in the present invention with or without targeting portions, since the positively charged backbone allows cellular internalization even in the absence of the BTX linkage. native, address or internalization domains. Alternatively, the botulinum toxin may be in the form of a toxin precursor botulinum, which by itself can be non-toxic, for example, a non-toxic zinc protease that becomes toxic at the time of proteolytic dissociation. The present invention also contemplates the general use of combinations and mixtures of botulinum toxins, although due to their different nature and properties, mixtures of botulinum toxin serotypes are generally not administered at this time. Similarly, the term "insulin" includes insulin extracted from natural sources, as well as insulin that can be obtained synthetically, through chemical or recombinant means. The insulin may also be in a modified form, or in the form of, for example, a recombinant peptide, a fusion protein or a hybrid molecule, or the insulin in a particular case may be a part of the insulin molecule that has the necessary activity. The same is true for other proteins that can be used in these particular transdermal compositions and methods, particularly antigens for immunization, which can vary widely in physiochemical properties. In the same way therapeutic agents without nucleic acid, without protein, including antifungal agents, can be obtained from natural sources or can be synthesized. The compositions of the present invention are preferably in the form of products that will be applied to the skin or epithelia of subjects or patients, i.e., humans or others. mammals that need treatment in particular. The term "they need" includes both pharmaceutical and health-related needs, as well as needs that tend to be more cosmetic, aesthetic or subjective. The botulinum toxin compositions can also be used, for example, to alter or improve the appearance of facial tissue. Through the use of the positively charged vehicles of the present invention, a botulinum toxin can be administered transdermally to a subject to treat conditions such as undesirable facial muscle spasm, as well as other muscle spasms, hyperhidrosis, acne conditions or any part of the body where the release of muscular ailments or spasms is desired. Botulinum toxin is administered topically for transdermal delivery to muscles or other structures associated with the skin. The administration can be performed, for example, on the legs, shoulders, back including lower back, armpits, palms, feet, neck, groin, dorsal of the hands or feet, elbows, upper arms, knees, upper part of the legs, buttocks, torso, pelvis or any other part of the body, where the administration of botulinum toxin is desired. The administration of botulinum toxin can be carried out to treat other conditions, including the treatment of neurological pain, prevention or reduction of headache by migraine or other headache, prevention or reduction of acne, prevention or reduction of dystonia or contractions dystonic, either subjective or clinics, prevention or reduction of symptoms associated with subjective or clinical hyperhidrosis, reduction of hypersecretion or sweating, reduction or increase of immune response, or treatment of other conditions for which the administration of botulinum toxin has been suggested or is carried out by injection. The administration of botulinum toxin, other therapeutic proteins that do not have a therapeutic effect on blood glucose levels, other antifungal agents useful for immunization described herein, or other therapeutic agents without protein, without nucleic acid, eg, botulinum toxin in complex, it can be carried out for purposes related to immunization. Alternatively, the complex can be prepared and applied topically to augment an immune response, for example, to provide immunizations with respect to various proteins, for example, infant immunizations in injections or immunization against various environmental hazards. Surprisingly, the administration of botulinum toxin or other therapeutic proteins described herein can also be carried out to reduce immune responses. The present invention allows the BTX or other protein to be delivered through an altered administration route and changes the presentation of the complex antigen of the agent and therefore may be useful for reducing immune response to antigens of said protein, and therefore facilitates repeated administration without reduction in activity related to the immune system. In general, the compositions are prepared by mixing the insulin, botulinum toxin or another biologically active agent such as, for example, a therapeutic protein which does not alter the therapeutic levels of glucose in the blood, a therapeutic nucleic acid-based agent, a therapeutic agent without nucleic acid, protein or alternatively an agent for immunization that will be administered with the positively charged vehicle, and typically with one or more additional pharmaceutically acceptable carriers or excipients. In their simplest form they may contain a pharmaceutically acceptable simple aqueous carrier or diluent, such as saline, which may be buffered. However, the compositions may contain other typical ingredients in topical pharmaceutical or cosmetological compositions, that is, a carrier, vehicle or dermatologically or pharmaceutically acceptable medium, i.e., a carrier, vehicle or medium that is compatible with the tissues to which it will be applied. The term "dermatologically or pharmaceutically acceptable", as used in the present invention, means that the compositions or components thereof already described, are suitable for use in contact with these tissues and for use in patients in general without toxicity, incompatibility , instability, undue allergic response and the like. As appropriate, the compositions of the present invention can comprise any ingredient that is conventionally used in the fields under consideration, particularly in cosmetology and dermatology. In all the aspects of the present invention, the association between the vehicle and the biologically active agent is by non-covalent interaction, which may include, for example, ionic interactions, hydrogen bonding, van der Waals forces or combinations thereof. The compositions can be pre-formulated or can be prepared at the time of administration, for example, by providing an assembly kit on or before the time of administration. Alternatively, as mentioned above, the botulinum toxin or other therapeutic protein and the skeleton or positively charged vehicle can be administered separately to the patient, for example, by providing a device containing a skin patch or other diagnostic device. a supply containing the therapeutic protein and a liquid, gel, cream or the like containing the positively charged vehicle (and optionally other ingredients). In that particular embodiment, the combination is administered by applying the liquid or other composition containing the vehicle to the skin, followed by application of the skin patch or other apparatus. The compositions of the present invention are applied to administer an effective amount of the insulin, botulinum toxin or other beneficial substance. For transdermal delivery, the term "effective amount" refers to any composition or method that provides greater transdermal delivery of the biologically active agent relative to the agent in the absence of the vehicle. For botulinum toxin, the term "effective amount" as used in the present invention means an amount of a botulinum toxin, as defined above, that is sufficient to produce the desired muscle paralysis or other effect, although it is implicitly a safe amount, that is, one that is low enough to avoid severe side effects. The desired effects include the relaxation of certain muscles with the help, for example, of diminishing the appearance of fine lines and / or wrinkles, especially on the face, or adjusting the facial appearance in other forms such as expanding the eyes, lifting the corners of the mouth, smoothing the lines that are marked from the upper lip or the general release of muscular tension. The aforementioned effect, the general release of muscle tension, can be achieved on the face or on any part of the body, for example, in the back or legs. For insulin, the term "effective amount" means in a similar manner an amount of insulin that is sufficient to produce the desired effect, i.e., decrease in glucose in the blood of the patient or subject. For antigens, the term "effective amount" refers to an amount sufficient to allow a subject to mount an immune response to the antigen after application or a series of applications of the antigen. For antifungal agents, the term "effective amount" refers to an amount sufficient to reduce symptoms or signs of fungal infection. For other biologically active agents that do not alter blood glucose levels in a therapeutic way, the term "amount "effective" refers to a quantity sufficient to exert the defined biological or therapeutic effect characterized by the agent in, for example, the Desk Reference of the Specialists or the like, without inducing significant toxicity The present invention specifically excludes antibody fragments They have no biological activity other than just binding a specific antigen when the term "therapeutic" or "biologically active protein" is used, since the antigens suitable for administration have other biological activities such as mounting an immune response, these remain included in the In addition, agents having a biological activity or the therapeutic effect through the binding of the specific antigen, thus blocking in ligand binding or altering the conformation of the antigen, are included in the present invention. The compositions may contain a suitable effective amount of insulin, botulinum toxin or other biologically active agent such as, for example, a therapeutic protein that does not alter in therapeutic form the levels of glucose in the blood, a therapeutic nucleic acid-based agent, a therapeutic agent without nucleic acid, without protein, or alternatively, an agent for immunization, for application in the form of a single dose treatment, or it can be more concentrated either for dilution at the site of administration or for use in multiple applications . In general, compositions containing botulinum toxin or other biologically active agent such as, for example, a therapeutic protein that does not alter in a therapeutic manner the blood glucose levels or a therapeutic nucleic acid-based agent, will contain from about 1 x 10"20 to about 25% by weight of the biologically active agent and from about 1 x 10"19 to about 30% by weight of the positively charged vehicle. In general, compositions containing a therapeutic agent without nucleic acid, without protein or, alternatively, an agent for immunization will contain from about 1 x 10"10 to about 49.9% by weight of the antigen and from about 1 x 10" 9 to about 50% by weight of the positively charged vehicle. In general, in a suitable form of application to the subject, the compositions of the present invention will contain from about 0.001 to about 10,000, preferably from about 0.01 to about 1000 lU / g of a composition comprising botulinum toxin and a charged carrier molecule in positive form as described in the present invention. The vehicle: botulinum toxin ratio preferably ranges from about 10: 1 to about 1.01: 1 and more preferably from about 6: 1 to about 1.15: 1 respectively. The amount of carrier molecule or the proportion thereof to the botulinum toxin, will depend on the vehicle chosen to be used in the composition in question. The appropriate amount or proportion of carrier molecule in a given case can be easily determined, for example, by carrying out one or more experiments such as those described below. The compositions of the present invention allow the delivery of a purer botulinum toxin with pharmacokinetics potentially enhanced by higher level specific activity. In addition, the positively charged vehicle reduces the need for the aforementioned accessory proteins, (e.g., human serum albumin ranging from 400-600 mg or recombinant serum albumin ranging from 250-500 mg) and stabilizers of polysaccharide and can produce beneficial reductions in immune responses for BTX. In addition, the compositions are suitable for use in physiological environments with a pH ranging from 4.5 to 6.3., and can therefore have said pH. The compositions may be stored preferably either at room temperature or under refrigeration conditions. Compositions or apparatus containing botulinum toxin, will generally be applied to provide the botulinum toxin in a dose from about 1 U to about 20,000 U, preferably from about 1 U to about 10,000 U, of botulinum toxin per cm2 of skin, per application. Larger doses within these ranges could be used preferably together with release materials controlled, for example, or leave a shorter retention time on the skin before being eliminated. In the case of insulin, the compositions of the present invention will contain from about 0.01 1 U to about 5000 U, preferably from about 0.1 U to about 500 U / gram. A composition comprising a form of insulin and a carrier molecule positively charged as described in the present invention, ranges from about 30: 1 to about 1.01: 1 and more preferably from about 6: 1 to about 1.25: 1. of insulin: transporter, respectively. Similarly, the amount of carrier molecule or the ratio of this to insulin, will depend on the vehicle chosen to be used in the composition in question. In terms of their form, the compositions of the present invention can include solutions, emulsions (including microemulsions), suspensions, creams, lotions, gels, powders or other typical solid or liquid compositions used for skin application, or other tissues where the compositions can be used. Said compositions may contain, in addition to the botulinum toxin, insulin or other biologically active agent, and the carrier molecule, other ingredients normally used in said products, such as antimicrobials, humectants and hydrating agents, penetrating agents, preservatives, emulsifiers, natural or synthetic oils, solvents, surfactants, detergents, gelatinization agents, emollients, antioxidants, fragrances, fillers, thickeners, waxes, odor absorbers, ink materials, coloring agents, powders, viscosity control agents and water, and optionally include anti-itch active anesthetics, botanical extracts, conditioning agents, darkening or brightening agents, brighteners, humectants, mica, minerals, polyphenols, silicones or derivatives thereof, sunblocks, vitamins and phytomedicines. In all aspects of the present invention, the association between the carrier and the biologically active agent is by non-covalent interaction, which may include, for example, ionic interactions, hydrogen bonding, van der Waals forces or combinations thereof. The compositions according to the present invention may be in the form of controlled release or sustained release compositions, wherein the insulin, botulinum toxin or other substance to be delivered and the vehicle, are encapsulated or otherwise contained within a material so that they are released into the skin in a controlled manner over time. The substance to be delivered and the carrier can be contained within matrices, liposomes, vesicles, microcapsules, microspheres and the like, or within a solid particulate material, which are all selected and / or constructed to provide release of the substance or substances over time. The therapeutic substance and the vehicle can be encapsulated together (for example, in the same capsule) or separately (in separate capsules). The administration of the compositions of the present invention to a subject is, of course, another aspect of the present invention. In the case of botulinum toxin, most preferably the compositions are administered by or under the direction of a specialist or other health professional. They can be administered in a single treatment or in a series of periodic treatments over time. For the transdermal delivery of botulinum toxin for the aforementioned purposes, a composition, as described above in topical form to the skin, is applied at a location or locations where the effect is desired. Due to its nature, most preferably the amount of botulinum toxin applied should be applied with care, in a range of application and frequency of application that will produce the desired result without producing any adverse or undesired results. In the case of insulin, for hospitalized patients or in-office treatments, the administration will be carried out by or under the direction of a health care professional, although on the other hand, it is likely to be carried out through the patient. It is likely that administration by skin patches and the like, with controlled release and / or monitoring is a common method, so that the compositions containing insulin of the present invention will often be provided as contained in a skin patch or other apparatus. In the case of antigens suitable for immunization, most preferably the compositions are administered by or under the direction of a physician or other health professional. They can be administered in a single treatment or in a series of periodic treatments over time. Accordingly, sustained release compositions are also contemplated in the present invention. For the transdermal delivery of antigens suitable for immunizations for the aforementioned purposes, a composition as described above is applied topically to the skin or to a nail plate and surrounding skin. In the case of therapeutics without nucleic acid, without protein, such as antifungal agents, preferably the compositions are administered under the direction of a specialist or other health professional. They can be administered in a single treatment or in a series of periodic treatments over time. Sustained release compositions are also contemplated for therapeutics without nucleic acid, without protein. The antifungal agents can be administered to the finger nail or to the foot nail plate or surrounding anatomical structures using, for example, a prosthetic nail plate, a lacquer, or a nail polisher with a nail polish agent. color, a gel, or a combination of any or all of these. For transdermal supply of botulinum toxin, for the purposes before mentioned above is topically applied to the skin a composition as described above. Equipment for administering the compositions of the present invention, either under the direction of a health care professional or through the patient or subject, may also include a normal applicator suitable for that purpose. The term "normal applicator" includes the means mentioned above for administering antifungal agents. In another aspect, the present invention relates to methods for topical administration of the positively charged carrier combination described above, with an effective amount of insulin, botulinum toxin, antigens suitable for immunization, antifungal agents or other biologically active agent such as such as, for example, a therapeutic protein that does not alter the therapeutic levels of blood glucose, a therapeutic nucleic acid-based agent, or a therapeutic agent without nucleic acid, without protein in general. As described above, administration can be carried out through the use of a composition according to the present invention, which contains suitable types and amounts of these two substances, specifically vehicle and biologically active agent. However, the present invention also includes the administration of these two substances in combination, although not necessarily in the same composition. For example, the therapeutically or biologically active substance may be incorporated in dry form in a skin patch or other delivery apparatus, and the positively charged vehicle can be applied to the surface of the skin prior to application of the patch, so that the two act together resulting in the desired transdermal delivery. In this sense, the two substances, specifically the carrier and the biologically active agent, act in combination or together, or possibly interact to form a composition or combination in situ. Methods for Preparing Compositions In another aspect, the present invention provides a method for preparing a pharmaceutical composition, wherein the method comprises combining a positively charged skeletal component and at least two members selected from the group consisting of: i) a first a negatively charged skeleton having a plurality of adhered image generation portions, or alternatively, a plurality of negatively charged image generation portions; ii) a second negatively charged skeleton having a plurality of adhered steering agents, or alternatively, a plurality of negatively charged steering portions; iii) at least one selected member of RNA, DNA, ribozymes, modified oligonucleic acids and cDNA encoding a selected transgene; iv) DNA encoding at least one persistence factor; and v) a third skeleton loaded in a negative form having a plurality of biological agents adhered, or a biological agent loaded in a negative form; with a pharmaceutically acceptable carrier to form a non-covalent complex having a net positive charge, provided that at least one of the members is selected from i), ii), iii) or v). In a related aspect, as described in the present invention, in some embodiments or compositions of the present invention, the skeleton or vehicle positively charged can be used only to provide transdermal delivery of certain types of substances. Preferred herein are compositions and methods comprising a biologically active agent such as botulinum toxin or other therapeutic protein that does not decrease blood glucose containing from about 1 x 10"20 to about 25% by weight of the biologically active agent and from about 1%. x 10"19 to about 30% by weight of the positively charged vehicle. Also preferred are compositions and methods comprising a therapeutically without protein, without nucleic acid such as an antifungal agent or an antigen suitable for immunization containing from 1 × 10"to about 49.9% by weight of the antigen and from about 1 × 10. "9 to about 50% by weight of the positively charged vehicle. In all aspects of the present invention, the association between the vehicle and the biologically active agent is by non-covalent interaction, which may include, for example, ionic interactions, hydrogen bonding, van der Waals forces or combinations thereof. The broad applicability of the present invention is illustrated by the ease with which a variety of pharmaceutical compositions can be formulated. Typically, the compositions are prepared by mixing the positively charged backbone component with the desired components of interest (eg, DNA, targeting components, imaging or therapeutics) in proportions and a sequence to obtain compositions having a net charge positive variable. In many embodiments, the compositions can be prepared, for example, at the head using pharmaceutically acceptable carriers and diluents for administration of the composition. Alternatively, the compositions can be prepared by suitably mixing the components and subsequently lyophilizing and storing them (usually at room temperature or beyond) until they are used or formulated in a suitable delivery vehicle. The compositions can be formulated to provide mixtures suitable for topical, cutaneous, oral, rectal, vaginal, parenteral, intranasal, intravenous, intramuscular, subcutaneous, intraocular, transdermal, etc. administration. The pharmaceutical compositions of the present invention preferably contain a vehicle which is pharmaceutically acceptable for an injectable formulation, in particular for direct injection into the desired organ or for topical administration (to the skin and / or mucous membrane). In particular they can be sterile, isotonic solutions or dry compositions, in particular freeze-dried compositions, which, by adding, depending on the case, sterile water or physiological saline solution, allow the injectable solutions to be prepared. For example, the doses of nucleic acid used for the injection and the number of administrations can be adapted according to various parameters, and in particular according to the mode of administration used, the pathology concerned, the gene to be expressed, or as an alternative the desired duration of treatment. Alternatively, when the compositions are to be applied topically, for example, when transdermal delivery is desired, the component or components of interest can be applied in dry form to the skin, for example, using a skin patch, when the skin is treated separately with the skeleton or vehicle positively charged. In this form the general composition is formed essentially in situ and administered to the patient or subject. Methods for using the compositions Delivery methods The compositions of the present invention can be delivered to a subject, cell, target site, either in vivo or ex vivo using a variety of methods. In fact, any of the routes normally used to introduce a composition in final contact with the tissue to be treated can be used. Preferably, the compositions will be administered with pharmaceutically acceptable carriers. Suitable methods of administration, such as compounds are available and are well known to those skilled in the art, and although, more than one route can be used to administer a particular composition, often a particular route provides a more immediate reaction and more effective than another route. The pharmaceutically acceptable carriers are determined in part, through the particular composition being administered, as well as by the particular method used to administer the composition. Accordingly, there is a wide variety of suitable formulations of pharmaceutical compositions of the present invention (see for example the publication of Remington's Pharmaceutical Sciences, 17th edition, 1985). The administration can be, for example, intravenous, topical, intraperitoneal, subdermal, subcutaneous, transcutaneous, intramuscular, oral, intraunion, parenteral, intranasal, or by inhalation. Suitable administration sites therefore include, but are not limited to, the skin, bronchi, gastrointestinal tract, eyes and ears. The compositions typically include a conventional pharmaceutical carrier or excipient and may further include other medicinal agents, vehicles, adjuvants and the like.

Preferably, the formulation will be from about 5% to 65% by weight, of a composition of the present invention, the remainder being suitable pharmaceutical excipients. Suitable excipients can be designed for the composition and route of administration in particular by methods known in the art (see, for example, the publication REMINGTON'S PHARMACEUTICAL SCIENCES, 18th EDITION, Mack Pubiishing Co., Easton, PA (1990)). The formulations may take the form of solid, semi-solid, lyophilized, powdered or liquid forms of solidification, such as, for example, tablets, pills, capsules, powders, solutions, suspensions, emulsions, suppositories, retention enemas, creams, ointments, lotions. , gels, aerosols, or the like. In embodiments wherein the pharmaceutical composition takes the form of a pill, tablet or capsule, the formulation may contain, together with the biologically active composition, or any of the following: a diluent such as lactose, sucrose, dicalcium phosphate and the like; and a disintegrant such as starch or derivatives thereof; a lubricant such as magnesium stearate and the like; and a linker such as starch, gum acacia, polyvinyl pyrrolidone, gelatin, cellulose and derivatives thereof. The compositions may be presented in sealed unit dose or multiple dose containers, such as ampoules or flasks. The doses administered to a patient must be sufficient to achieve a beneficial therapeutic response in the patient with the weather. The present invention specifically excludes antibody fragments that have no biological activity in addition to binding only a specific antigen when the term "therapeutic" or "biologically active protein" is used. Since the antigens suitable for immunization have other biological activities, such as assembly of an immune response, they remain included in the appropriate aspects of the present invention. In addition, agents that have a biological activity or a therapeutic effect through the binding of a specific antigen, thereby blocking the ligand binding or altering the conformation of the antigen, are included in the present invention. In some embodiments, a sustained release or controlled release formulation can be administered to an organism or cells in culture and the desired compositions can be carried. The sustained release composition can be administered to the tissue of an organism, for example, by injection. By the term "sustained release" is meant that the composition, preferably one which encodes a transgene of interest or a biological or therapeutic agent, is made available for uptake by the surrounding tissue or cells in culture for a period of time greater than that it could be achieved by administering the composition in a less viscous medium, for example, a saline solution. The compositions, alone or in combination with other suitable components, can be prepared in formulations of aerosol (for example, they can be "nebulized") to be administered through inhalation. The aerosol formulations can be placed in acceptable pressurized propellants, such as dichlorodifluoromethane, propane, nitrogen and the like. For administration by inhalation, the compositions may also be administered in the form of a dry powder (see, for example, the publication by Nektar Therapeutics, San Carlos, CA). Formulations suitable for parenteral administration, such as, for example, by intravenous, intramuscular, intradermal and subcutaneous routes, include aqueous and non-aqueous sterile isotonic injection solutions, which may contain anti-oxidants, regulators, bacteriostats and melted solutions which render the formulation with the projected receptor blood, and sterile aqueous and non-aqueous suspensions which may include suspending agents, solubilizers, thickening agents, stabilizers and preservatives. Other methods of administration include, but are not limited to, administration using angioplastic balloons, catheters and gel formations. Methods for delivery by angioplastic balloon, catheter and gel formation are well known in the art. Image generation methods One skilled in the art will be able to understand that the compositions of the present invention can be designed for a variety of imaging uses. In one modality, Virtual colonoscopy can be carried out using the system for component-based image generation. Currently, virtual colonoscopy essentially involves infusing contrast into a colon and visualizing CT images, then reconstructing a 3-D image. Similar techniques can be used for MR. However, feces, mucous and air all serve as contrast barriers and can produce an artificial surface to the reconstruction of the colon wall. In addition to a cell-phone-directional contrast that can help overcome these barriers, it provides real wall reconstruction and helps avoid both false and false negatives. There are several ways in which the component base system can be applied here. The simplest, cationic efficiency structure can be applied with a single contrast agent, for example, CT, MR or optical. Therefore, the cell surface layer can be visualized and any irregularities or detailed obstructions in image reconstruction. However, the component-based system offers the additional option of adding a second specific agent. This agent may consist of a skeleton of cationic efficiency, a portion of different imaging generation and targeting components, for example targeting two antigens characteristic of colon cancer. The portions of image generation from simple to diagnostic can be selected so that one is a contrast CT and the other contrast MR, or so that both are MR contrast, being an agent T2 and the other one agent T1. In this form, the surface can be reconstructed as indicated above, and any specific regions for a tumor antigen can be visualized and placed in the original reconstruction. In addition, the therapeutic agents can also be incorporated into the targeted diagnostic system. They can apply similar strategies to regional enteritis and ulcerative colitis (and again combine it with therapy). Alternatively, the optical image generating portions and detection methods may be employed, for example, in the case of diagnosis and administration of melanoma, preferably together with a fluorescent image generation portion. The optical image generating agent can be selected, for example, from the group including Cy3, Cy3.5, Cy5, Cy5.5, C7, Cy7.5, Oregon green 488, Oregon green 500, Oregon green 514, green fluorescent protein, 6-FAM, Texas Red, Hex, TET, and HAMRA. EXAMPLES Example 1 This example illustrates a composition suitable for transdermal delivery of a very large complex, i.e., a plasmid containing the blue fluorescent protein (BFP) transgene, using a skeleton or positively charged vehicle of the present invention. Selection of quinolone: The skeleton loaded in positive form was assembled by covalently adhering - Gly3Arg7 to polylysine MW 150,000 through the carboxyl of the terminal glycine to the free amines of the glycine side chains at a saturation degree of 18% (ie, 18 out of 100 lysine residues covalently adhere to -Gly3Arg7). The modified skeleton was designated "KNR2", to denote a second size of the peptidyl vehicle. The control polycation was unmodified polylysine (designated "K2", Sigma Chemical Co., St. Louis, MO) of the same size and from the same batch. An additional control polycation, Superfect® (Qiagen), which is an activated dendrimer-based agent, was selected as a reference for high-level in vitro transfection ranges (eg, simultaneous positive control and reference for efficiency of the state of the art versus in vitro toxicity.Therapeutic agent selection: An 8-kilobase plasmid (template based on pSport, Gibco BRL, Gaithersburg, MD), which contains all the transgene of the blue fluorescent protein (BFP) and sequences, was used. of partial flanking driven by a cytomegalovirus (CMV) promoter.BFP serves as an identifiable marker for cells that have been transfected, subsequently transcribes and translates the gene and can be visualized directly (eg, without additional staining) under fluorescence microscopy. Therefore, only cells in which the complex has crossed both the plasma membrane and the nuclear membrane before delivery or payload, may have transgene expression. East Plasmid in particular has a molecular weight of approximately 2.64 million, and was therefore selected to evaluate the delivery of very large therapeutics through these complexes. Sample preparation: In each case, a kind of polycation was used to assemble a final complex that has an excess of positive charge. Although increasing the load density increases the size (for example, more skeletons are found per complex), the increase in density of efficiency factor per complex can compensate for these changes. Therefore, an optimum can occur in low proportions (eg, based on size) or in high proportions (eg, based on factor-density efficiency) and both are evaluated here with respect to KNR2. The optimal proportions for efficiency K2 and efficiency Superfect, were selected based on the recommendation of the manufacturers and previous reports regarding maximum efficiency. The dose of nucleic-therapeutic acid was standardized across all groups as was the total volume and final pH of the composition that will be evaluated in the cell culture. The following mixtures were prepared: 1) K2 at a loading ratio of 4: 1 to a 0.5 ng / ml solution of a plasmid expressing blue fluorescent protein driven by a CMV promoter. 2) KN R2 in a ratio of 1: 5: 1 to a solution of 0.5 ng / ml of a plasmid expressing blue fluorescent protein driven by a CMV promoter. 3) KNR2 in a ratio of 10: 1 to a 0.5 ng / ml solution of a plasmid expressing blue fluorescent protein driven by a CMV promoter. 4) KNR2 in a ratio of 4: 1 to a 0.5 ng / ml solution of a plasmid expressing blue fluorescent protein driven by a CMV promoter. 5) KNR2 in a ratio of 1.25: 1 to a 0.5 ng / ml solution of a plasmid expressing blue fluorescent protein driven by a CMV promoter. 6) Superfect according to the manufacturer's recommendation in a 5: 1 charge ratio to a 0.5 ng / ml solution of a plasmid expressing blue fluorescent protein driven by a CMV promoter. Cell culture protocols: All cell culture experiments were carried out by blind observers for the identity of the treatment groups. In a six-well plate, 1.0 ml of each solution was added to 70% confluent HA-VSMC primary human aortic smooth muscle cells (passage 21: ATCC, Rockville, MD) and grown in M-199 with 10% serum. for 48 hours at a temperature of 37 ° C and with 10% CO2. The untreated control deposits were evaluated and each group was evaluated at n = 5 deposits per group.

Efficiency analysis: Photographs were obtained with low magnification (10X total) of intact cell plates, by blind observers at 60 degrees, 180 degrees, and 200 degrees from the top of each reservoir using a Nikon E600 epi-fluorescence microscope with a BFP filter and apocromat lenses. Image Pro Plus 3.0 image analysis adaptation (Media Cybernetics, Silver Spring, MD) was used to determine the percentage of total cell area that was positive. This result was normalized for the total cell area of each, and was reported as the gene delivery efficiency (percentage of total cells expressing the transgene at detectable levels). Toxicity analysis: Deposits were subsequently evaluated by blind observers in an ink exclusion test (viable cells exclude ink, while non-viable cells can not do so) followed by solubilization in 0.4% SDS in regulated saline by phosphate. Samples were evaluated in a Spectronic Genesys 5 UV / VIS spectrophotometer at 595 nm wavelength (blue) to quantitatively assess non-viable cells as a direct measure of the toxicity of the transfection agent. Samples were standardized to identical cell numbers by adjusting concentrations to correspond to OD280 values before OD595 measurements.

Data management and statistical analysis: The total positive staining was determined by blind observer through a batch image analysis using the Image Pro Plus software (Media Cybernetics, Silver Spring, MD) and normalized to total cross-sectional area for Determine the percentage of positive staining of each. Subsequently, the average and standard errors of each group were determined with 95% significance analysis of certainty in a form of repeated ANOVA measurements using the Staview software (Abacus, Berkeley, CA). Results: Efficiencies: The efficiency results are as indicated below (average + standard error): 1) 0.163 + 0.106% 2) 10.642 + 2.195% 3) 8.797 + 3.839% 4) 15.035 + 1.098% 5) 17.574 + 6.807 % 6) 1 .1 99 + 0.573% The runs no. 4 and no. 5 exhibit an increase in efficiency of statistically significant gene delivery (P <0.05 by a factor of repeated measures ANOVA with Fisher PLSD and TUKEY-A test) in relation to both polylysine alone and Superfect.

Toxicities: The average toxicity data are as indicated below (reported in AU at OD595, low values, as found with saline alone, correlate with low toxicity, although higher values, as found in the condition 1, indicate a high cellular toxicity: Saline - 0.057 A; 1) 3.460 A; 2) 0.251 A; 3) 0.291 A; 4) 0.243 A; 5) 0.297 A; 6) 0.337 A. Conclusions: A more efficient, less toxic gene delivery with a ratio of 1.25 to 4.0 of KNR2 to DNA can be achieved than controls, even those with a normal Superfect gold standard. This experiment confirms the ability to deliver very large therapeutic complexes through membranes using this vehicle. Example 2 This example illustrates the transport of a large nucleic acid through the skin through a vehicle of the present invention after a single administration.

Skeletal selection: The positively charged skeleton was assembled by covalent adhesion of -Gly3Arg7 to polylysine MW 150,000 through the carboxyl of the terminal glycine to the free amines of the side chains of lysine at a saturation level of 18% (by example, 18 out of 100 lysine residues adhere covalently to -Gly3Arg). The modified skeleton was designated "KNR2" as indicated above. The control polycation was unmodified polylysine (designated "K2", Sigma Chemical Co., St. Louis, MO) of the same size and from the same batch. An additional control polycation, Superfect (Qiagen), which is a dendrimer-based agent, was selected as a reference for higher transfection ranges (for example, positive and simultaneous control and reference for efficiency of state of the art versus in vitro toxicity). Therapeutic agent selection: For this experiment, an 8.5 kilobase plasmid (template based on pSport, Gibco BRL, Gaithersburg, MD) containing all of the transgene for beta-galactosidase (ßgal) of E. coli and the sequences of partial flanking driven through a cytomegalovirus (CMV) promoter. Here it serves as an identifiable marker for cells that have been transported, subsequently transcribes and translates the gene and can be visualized directly after staining specific for the external enzyme. Therefore, only the cells in which the complex has penetrated the skin and has reached the target cell and transubjected through both the plasma membrane and the nuclear membrane before the delivery of payload, it can have a transgene expression. This particular plasmid has a molecular weight of approximately 2,805,000. Sample preparation In each case, an excess of polycation is used to assemble a final complex that has an excess of positive charge. The optimal proportions for efficiency K2, efficiency KNR2, and efficiency Superfect, were selected based on the recommendation of the manufacturer and before the in vitro experiments to determine the maximum efficiency. The therapeutic dose-nucleic acid was standardized across all the groups as was the total volume and final pH of the composition that will be applied topically. The samples were prepared as follows: Group labeled AK1: 8 micrograms of βgalgal plasmid (p / CMV-sport-βga1) were mixed until homogeneous by final aliquot (eg, 8 micrograms total) and peptidyl K2 vehicle in a loading ratio of 4: 1, and diluted to 200 microliters with phosphate-buffered saline. The resulting composition was mixed to homogeneity with 1.8 ml of Cetaphil humidifier and aliquoted in 200 microlitre portions for live experiments. Group labeled AL1: 8 micrograms of βgalgal plasmid (p / CMV-sport-ßga1) were mixed until homogeneity was achieved by final aliquot (e.g., 8 micrograms total) and K2 at a loading ratio of 4: 1, and diluted to 200 microliters with phosphate-buffered saline. The resulting composition was mixed to homogeneity with 1.8 ml of Cetaphil and aliquoted in 200 microliter portions for in vivo experiments. Group labeling AM 1: 8 micrograms of ßgal plasmid (p / CMV-sport-ßga1) were mixed until final homogeneity was obtained by final aliquot (for example, 80 micrograms total) and Superfect in a loading ratio of 5: 1, and diluted to 200 microliters with phosphate-regulated saline. The resulting composition was mixed to homogeneity with 1.8 ml of Cetaphil and aliquoted in 200 microliter portions for in vivo experiments. Animal experiments to determine transdermal delivery efficiency after a single treatment with peptidyl and therapeutic nucleic acid vehicles: Animals were anesthetized by inhalation of isoflurane during the application of the treatments. After being anesthetized, 6 C57 black mice (n = 4 per group) had measured dose of 200 microliters of the appropriate treatment applied to the cranial part of the dorsal back skin (selected because the mouse can not reach this region with its snout or legs). The animals did not undergo hair removal treatment. The animals recovered in a controlled heat environment to avoid hypothermia and once they responded, they were given food and water ad libitum at night. 24 hours after treatment, the mice were euthanized through CO2 inhalation, and the treated skin segments were collected with a full thickness by blind observers. The treated segments were divided into three equal parts, the skull part was fixed in 10% neutral formalin regulated for 12 to 16 hours, then stored in 70% ethanol until paraffin embedded. The central part was frozen in halves and used directly to stain beta-galactosidase at a temperature of 37 ° C in sections, as described previously (Waugh, JM, Kattash M., J. Li, E. Yuksel, MD Kuo, M. Lussier, AB Weinfeld, R. Saxena, ED Rabinovsky, S. Thung, SLC Woo, and SM Shenaq, Local Overexpression of Tissue Plasminogen Activator to Prevent Arterial Thrombosis in an in vivo Rabbit Model, Proc. Natl. Acad Sci. USA, 1999 96 (3): 1065-1070 Also: Elkins CJ, Waugh JM, Amabile PG, Minamiguchi H, Uy M, Sugimoto K, Do YS, Ganaha F, Razavi MK, Dake MD. to evaluate and Iimit in-stent restenosis Tissue Engineering June 2002; 8 (3): 395-407). The treated caudal segment was frozen split in two for solubilization studies. Toxicity: Toxicity was evaluated by ink exclusion on sections matched with those analyzed with respect to efficiency above. The sections only went through staining either for efficiency or for toxicity, since the methods are not reliably used together. For toxicity analysis, the sections were immersed in exclusion ink for 5 minutes, subsequently incubated at a temperature of 37 ° C for 30 minutes in 1% CO2. Any cells that did not exclude ink in this period of time were considered non-viable. Data management and statistical analysis: Data collection and image analysis were carried out by blind observers. The stained sections as indicated above, were photographed in full in a Nikon E600 microscope, with plan-apocromat lenses. The resulting images were processed by batch image analysis using Image Pro Plus software as indicated above, with manual confirmation to determine the positive number for beta-galactosidase enzyme activity (blue with the substrate method used here) or cellular toxicity. These results were normalized for total cross-sectional cell numbers by nuclear fast red staining for each, and tabulated as percentage of cross-section positive spotting. Subsequently, the average and standard errors for each group were determined with analysis of importance to 95% certainty in a form of repeated measures ANOVA using Stratview software (Abacus, Berkeley, CA). Results: The results were summarized in the table that is and illustrated in Figure 3. The transdermally charged peptidyl delivery vehicle in positive form achieved statistically significant increases in the efficiency of transgene delivery and expression versus both K2 (essentially negative control) and the comparison standard for efficiency, Superfect . Although Superfect did not achieve statistically significant improvements with respect to K2, KN R2 had more than one order of magnitude improvement in supply efficiency versus Superfect, in this model system. Example 2: Average and standard error for positive beta-galactosidase cells as a percentage of total number of treatment group.

P = 0.0001 (critical to 99%). The results for toxicity are presented in Figure 4, which illustrates the percentage of total area that remained non-viable 24 hours after treatment. Here, K2 exhibits statistically significant cellular toxicity relative to KN R2 or Superfect, even at a dose where K2 has a low transfer efficiency, as described above (Amabile, P.

G., J. M. Waugh, T. Lewis, C. J. EIkins, T. Janus, M. D. Kuo, and M. D. Dake. I ntravascular Ultrasound Enhances in vivo Vascular Gene Delivery. J. Am. Col. Cardiol. June 2002; 37 (7): 1975-80). Conclusions: The transdermal peptidyl vehicle can transport large complexes through the skin with high efficiencies, particularly due to the restrictions of transgene expression and total complex size described above. Here, the positive area, instead of the positive number, was used for analysis since (1) the method is highly simplified and has greater precision in image analysis, (2) the peak demonstrations of efficiencies had already been produced in II. . B forcefully, (3) area measurements provide a greater scope for understanding in vivo results, since non-cellular components occupy a substantial part of the cross-section, and (4) comparison with vehicle complexes without peptidyl further Large was facilitated. Example 3 This example illustrates the transdermal delivery of a large nucleic acid-based therapeutic through the skin, using a positively charged peptidyl vehicle of the present invention in seven daily applications in sequences. Skeletal selection: The positively charged peptidyl skeleton was assembled by covalent addition of -Gly3Arg7 to polylysine MW 150,000, through the carboxyl of the terminal glycine to free amines of the side chains of lysine at a saturation level of 18% (For example, 18 out of 100 lysine residues are covalently attached to -Gly3Arg7). The modified skeleton was designated "KNR2". The control polycation was unmodified polylysine (designated "K2", Sigma Chemical Co., St. Louis, MO) of the same size and from the same batch. Therapeutic batch selection: For the present experiment, an 8.5-kilobase plasmid (template based on pSport, Gibco BRL, Gaithersburg, MD) containing all the transgene for beta-galactosidase (ßgal) from E. coli and sequences of partial flanking driven through a cytomegalovirus (CMV) promoter. This particular plasmid has a molecular weight of about 2,805,000, and was therefore selected to evaluate the delivery of very large therapeutics through the skin through the peptidyl vehicles. Sample preparation: In each case, an excess of polycation was used to assemble a final complex having an excess of positive charge. The experimental proportions were selected to be parallel to the single dose experiments presented in the previous experiment. The dose of therapeutic-nucleic acid was standardized across all groups as was the total volume and final pH of the composition that will be applied topically. The samples were prepared as follows: Group labeled AK1: 8 micrograms of βgalgal plasmid were mixed until homogeneous (p / CMV-sport-ßga1) by final aliquot (e.g., 240 micrograms total) and peptidyl vehicle KNR2 at a loading ratio of 4: 1, and diluted to 600 microliters with phosphate-buffered saline. The resulting composition was mixed to homogeneity with 5.4 ml of Cetaphil and aliquoted in 200 microliter portions for in vivo experiments. Group labeled AL1: 8 micrograms of βgalgal plasmid (p / CMV-sport-ßga1) were mixed until homogeneous by final aliquot (for example, 240 micrograms total) and K2 in a loading ratio of 4: 1, and diluted to 600 microliters with phosphate-regulated saline. The resulting composition was mixed to homogeneity with 5.4 ml of Cetaphil and aliquoted in 200 microlitre portions for in vivo experiments. Animal experiments to determine the cumulative transdermal delivery efficiencies after 7 treatments once a day with peptidyl vehicle and nucleic acid therapeutics: The animals were anesthetized by inhalation of sofluorane during the application of the treatments. After being anesthetized, 6 C57 black mice (n = 4 per group) had measured dose of 200 microliters of the appropriate treatment applied to the cranial part of the dorsal back skin (selected because the mouse can not reach this region with its snout or legs). The animals did not undergo hair removal treatment. The animals recovered in a controlled heat environment to avoid hypothermia and once they responded, they were given food and water ad libitum during the night. This procedure was repeated once a day at approximately the same time for 7 days. 7 days after the treatment, the mice were euthanized through CO2 inhalation, and the treated skin segments were collected with a total thickness by blind observers. The treated segments were divided into three equal parts, the skull part was fixed in 10% neutral formalin regulated for 12 to 16 hours, then stored in 70% ethanol until paraffin embedded. The central part was frozen split in two and used directly to stain beta-galactosidase at a temperature of 37 ° C in sections, as described above. The treated caudal segment was frozen split in two for solubilization studies. Data management and statistical analysis: Data collection and image analysis were carried out by blind observers. Sections stained as indicated above, were photographed in their entirety in a Nikon 600 microscope with plan-apocromat lenses. The resulting images were processed by batch image analysis using Image Pro Plus software as indicated above with manual confirmation to determine the positive area for beta-galactosidase enzyme activity. These results were normalized for the total cross-sectional area for each and tabulated as percentage of positive staining of cross section. Subsequently, the average and standard errors of each group were analyzed with importance analysis with 95% certainty in a form of measures repeated by ANOVA using the Statview software (Abacus, Berkeley, CA). Results The results are summarized in the table below and are illustrated in Figure 5. The transdermal peptidyl delivery vehicle achieved statistically significant increases in the delivery efficiency and expression of transgen versus K2. Example 3. Average and standard error for cumulative transgene expression of beta-galactosidase as percentage of total area after 7 applications of once a day for each treatment group.

P = 0.0012 (I mportance in 99%) Example 4 (vehicle without peptidyl) This example illustrates the transdermal delivery of a large nucleic acid-based therapeutic through the skin, using a positively charged peptidyl-free vehicle of the present invention in seven daily applications in sequences.

Skeletal selection: The positively charged skeleton was assembled by covalent adhesion - Gly3Arg7 to polyethyleneimine (PEI) MW 1, 000,000 through the carboxyl of the terminal glycine for free amines of PEI side chains at a saturation degree of 30% ( for example, 30 out of 1 00 lysine residues is covalently attached to a -Gly3Arg7). The modified skeleton was designated "PEI R" to denote the vehicle without large peptidyl. The control polycation was unmodified PEI (designated "PEI", Sigma Chemical Co., St. Louis, MO) of the same size and from the same batch. Selection of therapeutic agent For this experiment, an 8.5-kilobase plasmid (pSport-based template, Gibco BRL, Gaithersburg, MD) containing all beta-galactosidase transgene (ßgal) from E. coli and partial flanking sequences was used. operated through a cytomegalovirus (CMV) promoter. This particular plasmid has a molecular weight of approximately 2,805,000. Sample preparation In each case, an excess of polycation was used to assemble a final complex which has an excess of positive charge. The dose of therapeutic-nucleic acid was standardized across all groups, such as the total volume and final pH of the composition that will be applied topically. The samples were prepared as indicated below: AS labeled group: they were mixed until the homogeneity 8 micrograms of ßgal plasmid (p / CMV-sport-ßgal) per final aliquot (ie 240 micrograms total) and PEI control at a loading ratio of 5: 1, and diluted to 600 microliters with Tris-regulator EDTA The resulting composition was mixed until homogeneous with 5.4 ml of Cetafil and aliquoted in 200 microlitre portions for in vivo experiments. Labeling group AT: 8 micrograms of ßgal plasmid (p / CMV-sport-ßgal) were mixed until homogeneity was obtained by final aliquot (for example, 240 micrograms total) and vehicle without peptidyl compound PEIR ("PEI R") at a ratio of charge of 5: 1, and diluted to 600 microliters with Tris-EDTA regulator. The resulting composition was mixed until homogeneous with 5.4 ml of Cetafil and aliquoted in 200 microlitre portions for in vivo experiments. Group labeled AU: 8 micrograms of ßgal plasmid (p / CMV-sport-ßgal) were mixed until homogeneity was achieved by final aliquot (for example, 240 micrograms total) and vehicle without peptidyl Essentia purified with high level PEIR ("pure PEIR" ) at a loading ratio of 5: 1, and diluted to 600 microliters with Tris-EDTA buffer. The resulting composition was mixed until homogeneous with 5.4 ml of Cetafil and aliquoted in 200 microlitre portions for in vivo experiments. Animal experiments to determine cumulative transdermal supply efficiencies after 7 treatments at once to day with peptidyl-free and therapeutic nucleic acid vehicles: Animals were anesthetized by inhalation of isofuran during the application of the treatments. After being anesthetized, 6 C57 black mice (n = 3 per group) had measured doses of 200 microliters of the appropriate treatment applied to the cranial part of the dorsal back skin (selected because the mouse can not reach this region with the snout or leg). The animals did not undergo hair removal treatment. The animals were coated in a controlled warm environment to avoid hypothermia and once they responded they were given food and water overnight ad libitum. This procedure was repeated once a day at approximately the same time for 7 days. After 7 days of treatment, the mice were euthanized by CO2 inhalation, and segments of treated skin were collected in a full thickness by blind observers. The treated segments were divided into three equal parts, the cranial part was fixed in regulated formalin neutral at 10% for 12 to 16 hours, then stored in 70% ethanol until it was embedded in paraffin. The central part was thawed and used directly for beta-galactosidase staining at a temperature of 37 ° C in the sections previously described. The treated caudal segment was defrosted split in two for solubilization studies. Data management and statistical analysis: Data collection and image analysis was carried out by blind observers. The stained sections described above were photographed in full in a Nikon E600 microscope with plan-apochromat lenses. The resulting images underwent batch image analysis processing using Image Pro Plus software with manual confirmation to determine the positive area for beta-galactosidase enzyme activity. These results were normalized for the total cross-sectional area for each and tabulated as the percentage of positive cross-section staining. Subsequently, the average and standard error for each group was determined with analysis of importance to 95% certainty in a form of repeated ANOVA measurements using the Statview software (Abacus, Berkeley, CA). Results: The results are summarized in the table below and are illustrated in Figure 6. The transdermal delivery vehicle without peptidyl - both in compound form and in ultrapure form - achieved statistically significant increases in delivery efficiency and expression of transgen versus PEI. The ultrapure form of PEI R exhibited a tendency towards greater efficiencies than the standard PEIR in a manner consistent with the calculated greater specific activity of the reagent. Example 4. Average and standard error of cumulative transgene expression of beta-galactosidase in the form of percentage of total area after 7 applications once a day each treatment group.

P = 0.0058 (Importance in 99%) Conclusions: The transdermal vehicle without peptidyl can transport large complexes through the skin with greater efficiency, particularly due to the restrictions of transgene expression and the total complex size described above. Although the efficiencies are not as great as those obtained with smaller complexes of peptidyl vehicles, significant gains were achieved. It should be noted that the distribution of transgene expression using the large peptidyl-free complexes was almost exclusively based on the hair follicle, although the results for the peptidyl vehicles were diffuse in the cross sections. Therefore, the size and skeleton tropism can be employed for a nano-mechanical delivery direction. Example 5 This experiment demonstrates the use of a peptidyl vehicle to transport a large complex containing a botulinum toxin of intact tagged protein through intact skin after one-time administration relative to controls.

Skeletal selection: The skeleton loaded in positive form was assembled by covalent adhesion - Gly3Arg7 to polylysine MW 1 1 2,000 via carboxyl from the terminal glycine to free amines of the side chains of lysine at a saturation level of 1 8% ( for example, 1 8 out of 1 00 lysine residues adhere covalently to -Gly3Arg7). The modified skeleton was designated "KNR". The control polycation was unmodified polylysine (designated "K", Sigma Chemical Co., St. Louis, MO) of the same size and from the same batch. Therapeutic Agent: The Botox® brand of botulinum toxin A (Allergan) was selected for this experiment. It has a molecular weight of approximately 1 50,000. Sample preparation: The botulinum toxin was reconstituted according to the manufacturer's instructions. An aliquot of protein was biotinylated with a calculated 12-fold molar excess of biotin sulfo-NHS-LC (Pierce Chemical). The labeled product was designated as "Btox-b". In each case, an excess of polycation was used to assemble a final complex having an excess of positive charge such as in the delivery of high-level large negative nucleic acid complexes. A neutral or positive net charge avoids the repulsion of the protein complex of the high-level negative cell surface proteoglycans and extracellular matrix. HE standardized in all groups the dose of Btox-b, as the total volume and final pH of the composition that will be applied topically. The samples were prepared as follows: Group labeled "JMW-7": 2.0 units of Btox-b per aliquot (for example, 20 U total) and peptidyl vehicle KNR at a calculated MW ratio of 4: 1, they were mixed until homogeneous and diluted to 200 microliters with phosphate-buffered saline. The resulting composition was mixed until homogeneity was achieved with 1.8 ml of Cetafil and aliquoted in 200 microliter portions. Group labeled "JMW-8": 2.0 units of Btox-b were mixed until homogeneous, by aliquot (for example, 20 U total) and K in a loading ratio of 4: 1, and diluted to 200 microliters with solution Saline regulated by phosphate. The resulting composition was mixed until homogeneity was achieved with 1.8 ml of Cetafil and aliquoted in 200 microliter portions. Animal experiments to determine transdermal delivery efficiencies after a one-time treatment with labeled peptidyl and botulinum toxin vehicles: Animals were anesthetized through isofuran inhalation during the application of the treatments. After being anesthetized, 6 C57 black mice (n = 4 per group) underwent topical application of 200 microliters of the appropriate dose applied to the cranial part of the dorsal back skin (selected because the mouse can not reach this region with the snout or legs). The animals did not go through hair removal. At 30 minutes after the initial treatment, the mice were euthanized by CO2 inhalation, and the skin segments treated in a total thickness were collected by blind observers. The treated segments were divided into three equal parts; the cranial part was fixed in 10% neutral regulated formalin for 12 to 16 hours, then stored in 70% ethanol until paraffin embedded. The central part was thawed in two and used directly for biotin visualization by blind observers as summarized below. The treated caudal segment was thawed in two for solubilization studies. The biotin visualization was carried out as indicated below. In brief, each section was immersed for 1 hour in NeutrAvidin® regulation solution. To visualize the alkaline phosphatase activity, cross sections were washed with saline four times subsequently submerged in NBT / BCI P (Pierce Scientific) for 1 hour. The sections were then rinsed in saline and photographed in full on a Nikon E600 microscope with plan-apocromat lenses. Data management and statistical analysis: The total positive staining was determined by blind observer through batch image analysis using Image Pro Plus software (Media Cybernetics, Silver Spring, MD) and normalized to the total cross-sectional area to determine the percentage of positive staining for each one. Subsequently, the average and standard error of each group was determined with analysis of importance to 95% certainty in a form of repeated ANOVA measurements using the Statview software (Abacus, Berkeley, CA). Results: The positive average cross-sectional area for biotinylated botulinum toxin was reported as percentage of total area after topical one-time administration of Btox-b either with KNR ("EB-Btox") or K ("ni") . The results are presented in the following table and illustrated in figure 7. In figure 7, the positive area for labeling was determined as a percentage of total area after three days of treatment once a day with "EB-Btox" , which continued with Btox-b and the peptidyl vehicle KN R and "ni", which contained Btoxb with polycation K as a control. They are illustrated for each group of average and standard error. Example 5. Average and standard error for botulinum toxin area labeled as percentage of total cross-section after topical one-time administration of Btox-b with KN R (JMW-7) or K (J MW-8) for 30 minutes .

P = 0.0001 (I mportance in 99%) EXAMPLE 6 Example 5 demonstrated that the transdermal peptidyl vehicle allowed cient transfer of botulinum toxin after topical administration in a murine model of intact skin. However, this experiment does not indicate whether the botulinum toxin was released from complex protein in a functional form after translocation through the skin. The following experiment was constructed to assess whether botulinum toxin can be delivered in therapeutic form through intact skin as a topical agent using this peptidyl vehicle (again, without covalent modification of the protein). The positively charged skeleton was again assembled by covalent adhesion of -GIy3Arg7 to polylysine MW 1 12,000 via carboxyl of the terminal glycine for free amines of the lysine side chains at a saturation degree of 18% (e.g. of every 100 lysine residues is covalently adhered to -Gly3Arg). The modified skeleton was designated as "KNR". The control polycation was unmodified polylysine (designated "K", Sigma Chemical Co., St. Louis, MO) of the same size and from the same batch. The same botulinum toxin therapeutic agent was used as in Example 5, and was prepared in the same manner. Samples were prepared as follows: Group labeled "JMW-9": 2.0 units of botulinum toxin were mixed until homogeneous by aliquot (for example, 60 U total) and peptidyl vehicle KNR in a proportion MW calculated 4: 1, and diluted to 600 microliters with phosphate-buffered saline. The resulting composition was mixed until homogenous with 5.4 ml of Cetafíl and aliquoted in 200 microliter portions. Group labeled "J MW-1 0": 2.0 units of botulinum toxin were mixed until homogeneous by aliquot (for example, 60 U total) and K in a loading ratio of 4: 1 and diluted to 600 microliters with solution Saline regulated by phosphate. The resulting composition was mixed until homogenous with 5.4 ml of Cetafil and aliquoted in 200 microliter portions. Group labeled "JMW-1 1": were diluted to 600 microliters with phosphate-regulated saline solution, 2.0 units of botulinum toxin per aliquot (for example, 60 U total) without polycation. The resulting composition was mixed until homogenous with 5.4 ml of Cetafil and aliquoted in 200 microliter portions. Animal experiments to determine the therapeutic cacy after a one-time treatment with peptidyl and botulinum toxin vehicles: Animals were anesthetized by inhalation of isofuran during the application of the treatments. After being anesthetized, 6 C57 black mice (n = 4 per group) underwent topical application of 400 microliters of the appropriate treatment applied uniformly from the toes of the legs to the middle part of the thigh. Both extremities were treated, and treatments were randomized to either side. The animals did not go through hair removal. At 30 minutes after the initial treatment, the mice were evaluated for digital abduction capacity according to published digital abduction markers for leg mobility after administration of botulinum toxin (Aoki, KR.) A comparison of the margins of botulinum neurotoxin serotypes A, B and F in mice, Toxicon, 2001 Dec; 39 (12): 1815-20). Mobility of the mouse was also evaluated subjectively. Data management and statistical analysis: Digital abduction markers were tabulated independently through two blind observers. Subsequently the error was determined by means and standard of each group with analysis of importance to 95% certainty in a form of repeated measures ANOVA using the software Statview (Abacus, Berkeley, CA). Results: In the following table, digital abduction markers were presented after topical administration of botulinum toxin with KNR ("JMW-9"), K ("JMW-10") or diluent without polycation. ("JMW-1 1"), and are illustrated in the representative photomicrograph of Figure 8. The peptidyl vehicle KNR, produced statistically significant functional delivery of botulinum toxin through the skin in relation to both controls, which were comparable each. The Additional independent repeats (total of three independent experiments, all with identical conclusions on statistically significant paralysis of topical botulinum toxin with KNR, but no controls) from the present experiment confirmed these findings and did not reveal significant differences between topical botulinum toxin with or without K (per example, both controls). Interestingly, the mice consistently ambulated towards a paralyzed limb (which occurred in 100% of the treated animals and 0% of the controls of any control group). As shown in Figure 8, a limb treated with botulinum toxin in addition to the control polycation polylysine or botulinum toxin without polycation ("Btox alone") can mobilize digits (as a defense mechanism when it comes to lifting), although the extremities treated with botulinum toxin plus peptidyl vehicle KNR ("Essentia Btox lotion") can not be moved. Example 6. Digital abduction markers 30 minutes after the topical single-use of botulinum toxin with peptidyl vehicle KNR ("JMW-9 '"). with a control polycation K ("JMW-10") or only ("JMW-1 1 '").

P = 0.0351 (I mportance in 95%) Conclusions: This experiment serves to demonstrate that the transdermal peptidyl vehicle can transport a therapeutically effective amount of botulinum therapeutics through the skin without covalent modification of the therapeutic. The experiment also confirms that botulinum toxin does not work when applied topically in controls. Example 7 This experiment demonstrates the performance of a vehicle without peptidyl in the present invention. Skeleton selection: The positively charged skeleton was assembled by covalent adhesion -Gly3Arg7 for polyethyleneimine (PEI) MW 1, 000,000 through the carboxyl of the terminal glycine to free amines of the PEI side chains in a saturation degree of 30% (for example, 30 out of 100 lysine residues is covalently attached to -Gly3Arg7). The modified skeleton was designated "PEI R" to denote the vehicle without large peptidyl. The control polycation was unmodified PEI (designated "PEI", Sigma Chemical Co., St. Louis, MO) of the same size and from the same batch. The same botulinum toxin therapeutic agent as that of Example 5 was used. The botulinum toxin was reconstituted from the product BOTOX® according to the manufacturer's instructions. In each case, an excess of polycation was used to assemble a final complex that had an excess of positive charge as in the supply of high-level large negative nucleic acid complexes. A neutral or positive net charge prevents protein complex repulsion of proteoglycans from high-level negative cell surfaces and extracellular matrix. The dose of botulinum toxin was standardized across all groups as was the total volume and final pH of the composition that will be applied topically. The samples are prepared as follows: Group labeled "AZ": 2.0 units of botulinum toxin were mixed until achieving homogeneity by aliquot (for example 60 U total) and vehicles without peptidyl PEIR in an ultrapure form in a proportion of MW calculated at 5: 1 and diluted to 600 microliters with phosphate-buffered saline. The resulting composition was mixed until homogenous with 5.4 ml of Cetafil and aliquoted in 200 microliter portions. Group labeled "BA": 2.0 units of botulinum toxin were mixed until achieving homogeneity by aliquot (for example 60 U total) and PEI in a loading ratio of 5: 1 and diluted to 600 microliters with phosphate-regulated saline. The resulting composition was mixed until homogeneous with 5.4 ml of Cetafil and aliquoted in 200 microliter portions. Animal experiments to determine the therapeutic efficacy after the treatment of a single dose: Animals were anesthetized by isofuran inhalation during the application of the treatment. After being anesthetized, Six C57 black mice (n = 3 per group) underwent topical application of a 400-microliter dose of the appropriate treatment applied uniformly from the toes to the middle of the thigh. Both extremities were treated, and the treatments were randomized to either side. The animals did not go through hair removal. At 30 minutes after the initial treatment, the mice were evaluated by digital abduction capacity according to the digital abduction markers published for foot mobility after the administration of botulinum toxin (Aoki, KR). A comparison of the safety margins of botulinum neurotoxin serotypes A, B and F in Toxic mice. 2001 Dec; 39 (12): 1815-20). Mobility of the mouse was also evaluated subjectively. Data management and statistical analysis: Digital abduction markers were tabulated independently through two blind observers. The average and standard errors for each group were subsequently determined with analysis of importance to 95% certainty in a form of repeated ANOVA measurements using the Statview software (Abacus, Berkeley, CA). Results: Average digital abduction markers after topical one-time administration of botulinum toxin with ultrapure PEI R ("AZ"), or PEI control polycation ("BA"), and repeat (a single independent repeat for this experiment ), they are presented in the tables below. The vehicle without peptidyl PEIR produced statistically significant functional supply of botulinum toxin through the skin in relation to the controls. As indicated above, we observed walking in circles to the animals, directing attention to the paralyzed limbs. Example 7. Repetition 1. Digital abduction markers 30 minutes after topical administration of botulinum toxin with ultrapure PEI R ("AZ"), or PEI control polycation ("BA"). The average and standard errors are presented.

P = 0.0002 (I mportance at 99%) Eiem pio 7. Reo ection 2. Markers of digital abduction 30 minutes after the topical administration of one single botulinum toxin with PEI R ult rapuro ("AZ1"). 0 PETI con trol polycation ("BA1"). The average and standard errors are presented.

P = 0.0001 (importance in 99%) Conclusions: This experiment showed that the transdermal vehicle without peptidyl can transport therapeutic doses of botulinum toxin through the skin without prior covalent modification of the botulinum toxin. These discoveries complement those made with peptidyl transfer agents. The option of using a vehicle without peptidyl or peptidyl to achieve the therapeutic effect, will allow to design specific circumstances, environments and application methods and add to the scope of the transdermal delivery platform of the present invention. In these examples, the penetration of botulinum toxin either with peptidyl vehicles or without peptidyl versus topical botulinum toxin in the vehicle, further establishes the utility for the transdermal penetration of antigens for immunization, particularly for immunization with antigens that traverse the skin. poorly, such as botulinum. The supply of a botulinum toxin works to ensure that at least four different epitopes have been delivered, transdermal in an intact state; the fact that the botulinum toxin was not delivered in the absence of the vehicle in any example confirms that the vehicle produces potential significant immunization relative to the agent in the absence of the vehicle. Since immunization requires the antigens to pass through the skin in an amount sufficient to provide an immune response, this method allows the transdermal delivery of an antigen for immunization. Since this method does not require covalent modification of the antigen you do not need to understand the viral gene transfer, a number of advantages in terms of safety, stability and efficiency. Example 8 This experiment details the production of peptidyl vehicles and without peptidyl with TAT efficiency factors, as well as the assembly of these vehicles with botulinum toxins. Polyethylene imine coupling (PEI) to TAT fragment GGGRKKRRQRRR: The TAT fragment GGGRKKRRQRRR (6 mg, 0.004 mmol, Sigma Genosys, Houston, TX), which lacks all the side chain protection groups, was dissolved in 1 ml of 0.1 M MES regulator. To this was added EDC (3 mg, 0.016 mmol) followed by 50% solution with molecular weight of PEI 400k (p: v) in water, (-0.02 ml, -2.5 x 10-5 mmol). The pH was determined for 7.5 by test paper. Another 1 ml portion of 0.1 M MES was added and the pH adjusted to -5 by the addition of HCl. Another part of EDC (5 mg, 0.026 mmol) was added and the reaction, pH ~ 5 was stirred overnight. The next morning, the reaction mixture was frozen and lyophilized. Paste was made in 1 x sterile PBS, one column (diameter 1 cm x 14 cm height) of Sefadex G-25 (Amersham Biosciences Corp., Piscataway, NJ). The column was standardized by elution of FITC dextrans (Sigma, St Louis, MO) having a molecular weight of 19 kD. The standard was eluted initially in 5 ml of PBS, had an average peak at 6 ml and was queued in 7 ml. The above lyophilized reaction mixture was dissolved in a small volume of PBS and applied to the column. It was eluted by successive applications of 1 ml of PBS. The fractions were collected, the first of the first 3 ml eluted, including the reaction volume. The subsequent reactions were 1 ml. The eluted fractions were tested for UV absorbance in 280 nm. Fractions 3, 4 and 5 corresponding to 5 to 7 ml defined a modest absorbance peak. All fractions were lyophilized and the IR spectra were taken. The characteristic triple guanidine peak (2800-3000 cm-1) of the TAT fragment was observed in fractions 4 through 6. These fractions also showed an amide stretch of 1700 cm-1, thus confirming the conjugate of the TAT fragment and PEÍ. Another interaction was run using the TAT fragment GGGRKKRRQRRR (1.6 mg, 0.007 mmol). This amount was calculated so that one in 30 of the PEI amines can be reacted with the TAT fragment. The composition of the original polylysine-oligoarginine (KNR) efficiency factor described above is close. Successful covalent adhesion of the TAT fragment to the PEI amines was confirmed by IR as indicated above. Coupling of Polylysine to Fragment TAT To a solution of polylysine (10 mg 1.1 x 10-4 mmol; Sigma) in 1 ml of 0.1 M MES, pH -4.5, was added TAT fragment (4 mg, 0.003 mmol) subsequently EDC (3.5 mg, 0.0183 mmol). The resulting reaction mixture (pH ~ 4.5) was stirred at RT. The reaction it froze at a temperature of -78 ° C during the night. The next day the reaction mixture was thawed at RT and the pH was adjusted to -8 through the addition of saturated sodium bicarbonate. The reaction mixture was applied directly and to a Sephadex G-25 column constituted and standardized as described above. It was eluted in seven fractions of 1 ml and then 5 ml. The UV absorbance 280 was taken, revealing a relative peak in fraction 2,3 and 4. The IR of the lyophilized fractions revealed the characteristic guanidine peak (2800-300 cm -1) in fractions 1-7. Fraction 1 had a strong peak at 1730 cm-1 and nothing at 1600 cm-1, for fractions 2-6 the opposite was real. Therefore, successful covalent adhesion of the TAT fragment to a peptidyl vehicle, polylysine, was confirmed. The covalently attached TAT fragment and PEI (PEIT) and the covalently bound TAT fragment and polylysine (KNT) were subsequently mixed with botulinum toxin to form a non-covalent complex as follows: Group labeled "JL-1" " Mixed to achieve homogeneity 2..0 units of Btox-b per aliquot (for example, 20 U total) and PEIT at a loading ratio of 4: 1, and diluted to 200 microliters with phosphate-buffered saline. Group marked "JL-2" 2.0 units of Btox-b were mixed until homogeneous, by aliquot (for example, 20 U total) and KNT at a loading ratio of 4: 1 and diluted to 200 microliters with regulated saline by phosphate.

After formation of the non-covalent complex, the particles were centrifuged at 12,000 x g in a rotary microcentrifuge for 5 minutes, then resuspended in 20 microliters of deionized water and evaporated in a total reflectance cell attenuated by Germanium for IR. In this way the presence of Btox-b in the complexes was confirmed. In general, this experiment confirmed that synthetic schemes can be applied to other factors with efficiency and that the resulting vehicles can be made with a biologically active agent - in this case botulinum toxin - as in previous examples using vehicles with branching or branching groups. efficiency charged in a positive form of oligoarginine. Example 9 This experiment demonstrates the performance of a peptidyl vehicle for the generation of a specific antigen image. In this example, the complexes of one of the peptidyl vehicles Essentia, KNR2, with portions of optical image generation and targeting melanoma by modified antibodies are suitable for the topical detection of melanoma. Skeletal selection: The positively charged peptidyl skeleton was assembled by covalent adhesion of - Gly3Arg7 to polylysine MW 150,000 via the carboxyl of the terminal glycine to the free amines of the side chains of lysine at a saturation level of 18% (for example, 18 out of 100 lysine residues stick to covalently to -GIy3Arg7). The modified skeleton was designated as "KNR2". The control polycation was unmodified polylysine (designated "K2", Sigma Chemical Co., St. Louis, MO) of the same size and from the same batch. A murine monoclonal antibody to a conserved human melanoma domain, ganglioside 2; (IgG3, US Biologicals, Swampscott, MA) adhered covalently to a short polyaspartate anion chain (MW 3,000) through EDC coupling as indicated above to generate a derivative antibody designated "Gang2Asp". In addition, an anionic imaging agent was designated using an oligonucleic acid such as a polyanion, wherein the sequence was ATGC-J (designated in the following "ATGC-J") where "J" represents a Texas Red fluorophore adhered to covalent form, (Sigma Genosys, Woodlands, TX). For this experiment, 6.35 micrograms of Gang2Asp were combined with 0.1712 micrograms of ATGC-J and subsequently made into compound with 17.5 micrograms of KNR2 in a total volume of 200 microliters of deionized water to achieve a final ratio of 5: 1: 1 :: KNR2: ATGC-J: Gang2Asp. The mixture was vortexed for 2 minutes. The resulting complexes were applied to slices of CelITek Human Melanoma hydrated and sliced from CelITek Cytokeratin control (SDL, Des Plaines, IL) and incubated for 5 minutes before the photographic evaluation of fluorescence distribution versus light field distribution of melanoma pigment in the same field. Additional controls without ATGC-J or without Gang2Asp were also employed. Results: The non-covalent complexes produced a distribution of the optical imaging agent that followed the tropism of the antibody derivative rather than the distribution of the complexes in the absence of the antibody. It should be noted that the complexes followed a distribution that coincided with the pigmented melanoma cells, as illustrated in Figure 9. Conclusions: This experiment demonstrates the production of a viable complex for transport through the skin and visualization of melanoma. through optical techniques using a vehicle suitable for topical delivery. Such a method can be used, for example, together with surgical margin-adjustment or can be used in routine melanoma investigations. Similar strategies can easily be employed for topical diagnoses of other skin-related conditions, as those skilled in the art can appreciate. Due to the high sensitivity of the optical image generation portions, an improved detection of these conditions can be promised significantly through these non-covalent complexes. It will be understood that the examples and embodiments described herein are for illustrative purposes only and that those skilled in the art will be able to suggest various modifications or changes, and that they are included within the spirit and scope of the invention. present application and the appended claims. All the publications, patents and patent applications mentioned herein are incorporated in their entirety to the present invention as a reference for all purposes.

Claims (240)

1. R E I V I N D I C A C I O N S 1 . A composition comprising a biologically active protein that does not alter blood glucose levels in a therapeutic manner and a vehicle comprising a positively charged skeleton that has positively charged branching groups adhered to and which is in a effective amount for transdermal delivery, wherein the association between the vehicle and the biologically active protein is non-covalent.
2. 2. A composition as described in claim 1, characterized in that the composition provides greater transdermal delivery of the biologically active protein relative to the agent in the absence of the vehicle. 3. A composition as described in claim 2, characterized in that the biologically active protein has therapeutic activity. 4. A composition comprising a biologically active agent without nucleic acid, without protein, and a carrier comprising a positively charged skeleton having positively charged branching groups adhered to, and which is in an effective amount for transdermal delivery , wherein the association between the vehicle and the biologically active agent is non-covalent. 5. A composition as described in claim 4, characterized in that the composition provides a greater transdermal delivery of the biologically active agent relative to the people in the absence of the vehicle. 6. A composition as described in claim 5, characterized in that the biologically active agent has a therapeutic activity. 7. A composition as described in claim 3, characterized in that the therapeutic protein has a molecular weight of at least 55,000 kD. 8. A composition as described in claim 1, characterized in that the backbone comprises a positively charged polypeptide. 9. A composition as described in claim 8, characterized in that the backbone comprises a polypeptide created in a positive form having a molecular weight of from about 1,000 to about 1,500,000. 1 0. A composition as described in claim 8, characterized in that the backbone comprises a positively charged polypeptide having a molecular weight of from about 25,000 to about 1,200,000. eleven . A composition as described in claim 8, characterized in that the skeleton has a positively charged polypeptide having a molecular weight of from about 1,000,000 to about 1,000,000. 12. A composition as described in the claim 8, characterized in that it comprises a positively charged polylysine. 13. A composition as described in claim 12, characterized in that the backbone comprises a positively charged polylysine having a molecular weight from about 10,000 to about 1,500,000. 14. A composition as described in claim 1, characterized in that the backbone comprises a polylysine created in positive form having a molecular weight from about 25,000 to about 1,200,000. 5. A composition as described in claim 12, characterized in that the backbone comprises a polylysine created in a positive form having a molecular weight from about 1,000,000 to about 1,000,000. 16. A composition as described in claim 1, characterized in that the backbone comprises a polymer without peptidyl positively charged. 7. A composition as described in claim 16, characterized in that the polymer backbone without peptidyl comprises a positively charged polyalkyleneimine. 18. A composition as described in claim 1, characterized in that the polyalkyleneimine is polyethyleneimine. 1 9. A composition as described in claim 1 8, characterized in that the polyethyleneimine has a molecular weight from about 1,000 to 2,500,000. 20. A composition as described in claim 1 8, characterized in that the polyethyleneimine has a molecular weight from about 1,000,000 to about 1,800,000. A composition as described in claim 18, characterized in that the polyethyleneimine has a molecular weight from about 500,000 to about 1,400,000. 22. A composition as described in claim 1, characterized in that the carrier comprises a positively charged polymer having positively charged branching groups selected independently from - (gly) n -? - (arg) n2 , HIV-TAT, and fragments thereof, and Antennapedia PTD and fragments or mixtures thereof, wherein the subscript n1 is an integer from 0 to approximately 20 and the subscript n2 is independently an integer non from approximately 5 to approximately 25. 23. A composition as described in claim 22, characterized in that branching groups loaded in positive form are independently selected from groups having the formula - (gly) n? - (arg) n2. 24. A composition as described in claim 23, characterized in that the subscript n 1 is an integer from about 1 to about 8. 25. A composition as described in claim 23, characterized in that the subscript n1 is an integer of from about 2 to about 5. 26. A composition as described in claim 23. , characterized in that the subscript n2 is a non-number from from about 7 to about 17. 27. A composition as described in claim 23, characterized in that the subscript n2 is a number from about 7 to about 13. 28. A composition as described in claim 22, characterized in that the branching groups are selected from HIV-TAT and fragments thereof. 29. A composition as described in claim 28, characterized in that the branched groups loaded in positively adhered form are HIV-TAT fragments having the formula (gly) p-RGRDDRRQRRR- (gly) g, (gly) P - YGRKKRRQRRR- (giy) q, or (gly) p-RKKRRQRRR- (gly) q, where the subscripts p and q are each independently an integer from 0 to 20. 30. A composition as described in the claim 22, characterized in that the branching group are Antennapedia PTD groups or fragments thereof. 31. A composition as described in claim 22, characterized in that the polymer charged in the form positive comprises a polypeptide. 32. A composition as described in claim 31, characterized in that the polypeptide is selected from polylysines, polyalgenins and polyornithines. 33. A composition as described in claim 32, characterized in that the polypeptide is a polylysine. 34. A composition as described in claim 22, characterized in that the polymer comprises a polymer without peptidyl positively charged. 35. A composition as described in claim 34, characterized in that the polymer without peptidyl comprises a positively charged polyalkyleneimine. 36. A composition as described in claim 35, characterized in that the polyalkyleneimine is a polyethyleneimine. 37. A composition as described in claim 4, characterized in that the backbone comprises a positively charged polypeptide. 38. A composition as described in claim 37, characterized in that the backbone comprises a positively charged polypeptide having a molecular weight of from about 1,000 to about 1,500,000. 39. A composition as described in claim 37, characterized in that the skeleton comprises a positively charged polypeptide having a molecular weight of from about 25,000 to about 1,200,000. 40. A composition as described in claim 37, characterized in that the backbone comprises a positively charged polypeptide having a molecular weight of from about 100,000 to about 1,000,000. 41 A composition as described in claim 37, characterized in that the backbone comprises a polylysine positively charged. 42. A composition as described in claim 41, characterized in that the backbone comprises a positively charged polylysine having a molecular weight of from about 1,000 to about 1,500,000. 43. A composition as described in claim 41, characterized in that the backbone comprises a positively charged polylysine having a molecular weight of from about 25,000 to about 1,200,000. 44. A composition as described in claim 41, characterized in that the backbone comprises a positively charged polylysine having a molecular weight of from about 1,000,000 to about 1,000,000. 45. A composition as described in claim 4, characterized in that the backbone comprises a polymer without peptidyl positively charged. 46. A composition as described in claim 45, characterized in that polymer skeleton without peptidyl comprises a positively charged polyalkyleneimine. 47. A composition as described in claim 46, characterized in that the polyalkyleneimine is a polyethyleneimine. 48. A composition as described in claim 47, characterized in that the polyethyleneimine has a molecular weight of from about 10,000 to about 2,500,000. 49. A composition as described in claim 47, characterized in that the polyethyleneimine has a molecular weight of from about 100,000 to about 1,800,000. 50. A composition as described in claim 47, characterized in that the polyethyleneimine has a molecular weight of from about 500,000 to 1, 400,000. 51 A composition as described in claim 4, characterized in that the carrier comprises a positively charged polymer that has adhered positively charged branching groups independently selected from - (gly) n? - (arg) n2, HIV- TAT and fragments thereof, and PTD Antennapedia and fragments or mixtures thereof, wherein the subscript n1 is an integer from 0 to approximately 20, and the subscript n2 is independently a whole non of from about 5 to about 25. 52. A composition as described in claim 51, characterized in that branching groups charged in positive form are independently selected from groups having the formula - (gly) n? - (arg) n2. 53. A composition as described in claim 52, characterized in that the subscript n1 is an integer of from about 1 to about 8. 54. A composition as described in claim 52, characterized in that the subscript n1 is an integer of from about 2 to about 5. 55. A composition as described in claim 52, characterized in that the subscript n2 is a non-number from about 7 to about 17. 56. A composition as described in claim 52, characterized in that the subscript n2 is a non-number from from about 7 to about 13. 57. A composition as described in claim 51, characterized in that the branching groups are selected from HIV-TAT and fragments. thereof. 58. A composition as described in claim 57, characterized in that the branching groups loaded in positively adhered form are HIV-TAT fragments, which have the formula (gly) p-RGRDDRRQRRR- (gly) q, (gly) p-YGRKKRRQRRR- (gly) q, or (gly) p-RKKRRQRRR- (gly) q, where the subscripts p and q are each independently an integer from 0 to 20. 59. A composition as described in claim 51, characterized in that the branching groups are Antennapedia PTD groups or fragments thereof. 60. A composition as described in claim 51, characterized in that the positively charged polymer comprises a polypeptide. 61 A composition as described in claim 60, characterized in that the polypeptide is selected from polylysines, polyarginines, polyornithines and polyhomoarginines. 62. A composition as described in claim 61, characterized in that the polypeptide is a polylysine. 63. A composition as described in claim 51, characterized in that the polymer comprises a simple polymer without peptidyl positively charged. 64. A composition as described in claim 63, characterized in that the polymer without peptidyl comprises a positively charged polyalkyleneimine. 65. A composition as described in claim 64, characterized in that the polyalkyleneimine is a polyethyleneimine. 66. A composition as described in claim 4, characterized in that it contains from about 1 x 10-20 to about 25% by weight of the biologically active agent and from about 1 x 10-19 to about 30% of the positively charged vehicle. 67. A controlled release composition as described in claim 4. 68. A composition as described in claim 1, characterized in that the biologically active protein is a botulinum toxin. 69. A composition as described in claim 68, characterized in that the botulinum toxin is selected from botulinum toxin serotypes A, B, C, D, E, F and G. 70. A composition as described in Claim 68, characterized in that the botulinum toxin comprises a botulinum toxin derivative. 71 A composition as described in claim 68, characterized in that the botulinum toxin comprises a recombinant botulinum toxin. 72. A device for administration to a subject of a composition as described in claim 1, characterized in that it comprises an apparatus for supplying the biologically active agent and a vehicle comprising a positively charged skeleton having attached branching groups. charged positively, and which is in an effective amount for transdermal delivery. 73. A kit as described in claim 72, characterized in that the biologically active agent is a botulinum toxin. 74. A kit as described in claim 72, characterized in that the composition is contained in an apparatus for administering the biologically active protein to a subject through the skin or epithelium. 75. A device as described in claim 74, characterized in that the device is a skin patch. 76. A kit for administering a biologically active protein to a subject, characterized in that it comprises an apparatus for delivering the biologically active protein in the skin or epithelium, and a composition comprising a positively charged vehicle and having attached charged branching groups in positive form independently selected from - (gly) ni- (arg) n2, HIV-TAT and fragments thereof, and PTD Antennapedia and fragments or mixtures thereof, wherein the subscript n1 is an integer from 0 to approximately 20, and the subscript n2 is independently an integer non from about 5 to about 25, wherein the association between the vehicle and the biologically active protein is non-covalent. 77. A device as described in claim 76, characterized in that the device is a skin patch. 78. A method for administering to a subject a biologically active protein which does not alter in a therapeutic manner the glucose levels in the blood, characterized in that it comprises applying topically in the skin or epithelium of the subject the protein together with an effective amount of a positively charged vehicle comprising a positively charged skeleton and having attached branching groups loaded in a positive way, where the association between the vehicle and the biologically active protein is non-covalent. 79. A method as described in claim 78, characterized in that the composition provides a greater transdermal delivery of the biologically active protein relative to the agent in the absence of the vehicle. 80. A method as described in claim 79, characterized in that the biologically active protein has therapeutic activity. 81. A method for administering to a subject a biologically active agent without nucleic acid, without protein, characterized in that it comprises topically applying the skin or epithelium of the subject the biologically active agent together with an effective amount of a positively charged vehicle that it comprises a positively charged skeleton having adhered positively charged branching groups, wherein the association between the carrier and the biologically active agent is non-covalent. 82. A method as described in claim 81, characterized in that the composition provides a greater transdermal delivery of the biologically active agent with relationship to the agent in the absence of the vehicle. 83. A method as described in claim 82, characterized in that the biologically active agent has therapeutic activity. 84. A method as described in claim 80, characterized in that the biologically active protein and the vehicle deliver the subject in a composition containing both components. 85. A method as described in claim 80, characterized in that the biologically active protein and the vehicle separately deliver the subject. 86. A method as described in claim 83, characterized in that the biologically active protein and the vehicle deliver the subject in a composition containing both components. 87. A method as described in claim 83, characterized in that the biologically active agent and the vehicle are delivered separately to the subject. 88. A method as described in claim 80, characterized in that the composition is a controlled release composition or a sustained release composition. 89. A method as described in claim 83, characterized in that the composition is a controlled release composition or sustained release composition. 90. A method as described in claim 80, characterized in that the therapeutic protein is a botulinum toxin. 91 A method as described in claim 90, characterized in that the botulinum toxin is selected from botulinum toxin serotypes A, B, C, D, E, F and G. 92. A method as described in the claim 90, characterized in that the botulinum toxin comprises a botulinum toxin derivative. 93. A method as described in claim 90, characterized in that the botulinum toxin comprises a recombinant botulinum toxin. 94. A method as described in claim 90, characterized in that the botulinum toxin was administered to provide an aesthetic benefit and / or to the subject. 95. A method as described in claim 90, characterized in that the botulinum toxin was administered to the subject for prevention or reduction of symptoms associated with muscle spasm or constriction. 96. A method as described in claim 90, characterized in that the botulinum toxin and the positively charged vehicle is administered topically to a site on the subject's face. 97. A method as described in claim 90, characterized in that the botulinum toxin and the positively charged vehicle is administered topically to a site in the subject other than the face. 98. A composition comprising an antigen suitable for immunization and vehicle comprising a positively charged skeleton having adhered positively charged branching groups and which are in an effective amount for transdermal delivery, wherein the association between the vehicle and the antigen is non-covalent. 99. A composition as described in claim 98, characterized in that the backbone comprises a polypeptide loaded in a positive form. 1 00. A composition as described in claim 99, characterized in that the backbone comprises a positively charged polypeptide having a molecular weight of from about 10,000 to about 1,500,000. 1 01. A composition as described in claim 99, characterized in that the backbone comprises a positively charged polypeptide having a molecular weight of from about 25,000 to about 1,200,000. 1 02. A composition as described in claim 99, characterized in that the backbone comprises a positively charged polypeptide having a molecular weight of from about 1,000,000 to about 1,000,000. 1 03. A composition as described in claim 99, characterized in that the backbone comprises a polylysine positively charged. 1 04. A composition as described in claim 103, characterized in that the backbone comprises a positively charged polylysine having a molecular weight of from about 10,000 to about 1,500,000. 105. A composition as described in claim 103, characterized in that the backbone comprises a positively charged polylysine having a molecular weight of from about 25,000 to about 1,200,000. 106. A composition as described in claim 103, characterized in that the backbone comprises a positively charged polylysine having a molecular weight of from about 100,000 to about 1,000,000. 107. A composition as described in claim 98, characterized in that the backbone comprises a polymer without peptidyl positively charged. 108. A composition as described in claim 107, characterized in that the polymer backbone without peptidyl comprises a positively charged polyalkyleneimine. 109. A composition as described in claim 108, characterized in that the polyalkyleneimine is a polyethyleneimine. 10. A composition as described in claim 109, characterized in that the polyethyleneimine has a molecular weight of from about 10,000 to about 2,500,000. 1 1 1 A composition as described in claim 1 09, characterized in that the polyethyleneimine has a molecular weight of from about 1,000,000 to about 1,800,000. 12. A composition as described in claim 10, characterized in that the polyethyleneimine has a molecular weight of from about 500,000 to about 1,400,000. 1 1 3. A composition as described in claim 98, characterized in that the carrier comprises a positively charged polymer that has positively charged branching groups adhered to independently selected from - (gly) ni- (arg) n2 , HIV-TAT and fragments thereof, and PTD Antennapedia and fragments and mixtures thereof, wherein the subscript n 1 is an integer from 0 to approximately 20, and the subscript n 2 is independently an integer non of approximately 5 up to about 25. 14. A composition as described in claim 13, characterized in that branching groups loaded in positive form are independently selected from groups having the formula (gly) ni- (arg) n2. 1. A composition as described in claim 1 14, characterized in that the subscript n 1 is an integer of from about 1 to about 8. 1 16. A composition as described in claim 1 14, characterized in that the subscript n 1 is an integer of from about 2 to about 5. 1 1 7. A composition as described in claim 14, characterized in that the subscript n2 is an integer of from about 7. to about 13. 1 1 8. A composition as described in claim 1 14, characterized in that the subscript n2 is a non-number of from about 7 to about 1 3. 1 1 9. A composition as described in claim 11, characterized in that the branching groups are selected from HIV-TAT and fragments thereof. 120. A composition as described in claim 1 1 9, characterized in that the branching groups charged in positive form are H IV-TAT fragments having the formula (gly) p-RGRDDRRQRRR- (gly) q. (gly) p-TGRKKRRQRRR- (gly) q, or (gly) p-RKKRRQRRR- (gly) q characterized in that the subscripts p and q are each independently an integer from 0 to 20. 121. A composition as described in claim 13, characterized in that the branching groups are Antennapedia PTD groups. 122. A composition as described in claim 1 1 3, characterized in that the positively charged polymer comprises a polypeptide. 123. A composition as described in claim 122, characterized in that the polypeptide is selected from polylysines, polyarginines and polioarnitines. 124. A composition as described in claim 123, characterized in that the polypeptide is a polylysine. 125. A composition as described in claim 13, characterized in that the polymer comprises a polymer without peptidyl positively charged. 126. A composition as described in claim 125, characterized in that the polymer without peptidyl comprises a positively charged polyalkyleneimine. 127. A composition as described in claim 126, characterized in that the polyalkyleneimine is a polyethyleneimine. 128. A composition as described in claim 98, characterized in that it contains from about 1 × 10 -10 to about 49.9% by weight of the antigen and from about 1 × 10 -9 to about 50% by weight of the loaded vehicle in Positive way. 129. A controlled release composition as described in claim 98. 130. A composition as described in claim 98, characterized in that the antigen is a botulinum toxin. 1 31. A composition as described in claim 1, characterized in that the botulinum toxin is selected from botulinum toxin serotypes A, B, C, D, E, F and G. 1 32. A composition as described in Claim 1 30, characterized in that the botulinum toxin comprises a botulinum toxin derivative. 1 33. A composition as described in claim 130, characterized in that the botulinum toxin comprises a recombinant botulinum toxin. 1 34. A composition as described in claim 98, characterized in that the antigen is suitable for childhood immunizations. 1 35. A device for administration to a subject of an antigen suitable for immunization, characterized in that it comprises an apparatus for delivering the antigen to the skin or epithelium and a composition as described in claim 98. 136. Such equipment and as described in claim 1, characterized in that it comprises a customary applicator. 137. A device as described in claim 135, characterized in that the composition is contained in an apparatus for administering a subject and an antigen suitable for immunization through the skin or epithelium. 1 38. A device as described in claim 137, characterized in that the device is a skin patch. 1 39. A kit for administration to a subject of an antigen suitable for immunization, characterized in that it comprises an apparatus for delivering the antigen suitable for immunization to the skin or epithelium and a composition comprising a positively charged carrier having positively charged branching groups selected independently of (gly) m1 - (arg) n2, HIV-TAT and fragments thereof, and PTD Antennapedia and fragments and mixtures thereof, wherein the subscript n1 is an integer from 0 to about 20, and the subscript n2 is independently an integer non of from about 5 to about 25, wherein the association between the vehicle and the antigen is non-covalent. 140. A device as described in claim 139, characterized in that the device is a skin patch. 141. A method for administering to a subject an antigen suitable for immunization, characterized in that it comprises applying topically to the skin or epithelium of the subject the antigen suitable for immunization together with an effective amount of a positively charged vehicle comprising a skeleton loaded in positive form that has positively charged branching groups adhered to, where the association between the vehicle and the antigen is non-covalent. 142. A method as described in claim 141, characterized in that the antigen suitable for immunization and the vehicle are administered to the subject in a composition that contains both components. 143. A method as described in claim 141, characterized in that the antigen suitable for immunization and the vehicle are administered separately to the subject. 144. A method as described in the claim 141, characterized in that the backbone comprises a positively charged polypeptide. 145. A method as described in claim 144, characterized in that the backbone comprises a polypeptide loaded in a positive form and having a molecular weight of from about 1 0,000 to about 1, 500, 000. 146. Such a method and as described in claim 144, characterized in that the backbone comprises a polypeptide loaded in positive form and having a molecular weight of from about 25,000 to 1, 200, 000. 147. A method as described in claim 144, characterized in that the backbone comprises a positively charged polypeptide and having a molecular weight of from about 1,000,000 to about 1,000,000. 148. A method as described in the claim 144, characterized in that the skeleton comprises a polylysine positively charged. 149. A method as described in claim 148, characterized in that the backbone comprises a polylysine positively charged and having a molecular weight of from about 10,000 to about 1,500,000. 150. A method as described in claim 148, characterized in that the backbone comprises a polylysine positively charged and having a molecular weight of from about 25,000 to about 1,200,000. 151 A method as described in claim 148, characterized in that the backbone comprises a polylysine positively charged and having a molecular weight of from about 100,000 to about 1,000,000. 152. A method as described in the claim 141, characterized in that the backbone comprises a polymer without peptidyl positively charged. 153. A method as described in the claim 152, characterized in that the polymer backbone without peptidyl comprises a positively charged polyalkyleneimine. 154. A method as described in the claim 153, characterized in that the polyalkyleneimine is a polyethyleneimine. 155. A method as described in claim 154, characterized in that the polyethyleneimine has a molecular weight of from about 10,000 to about 2,500,000. 156. A method as described in the claim 154, characterized in that the polyethyleneimine has a molecular weight of from about 100,000 to about 1, 800,000. 1 57. A method as described in claim 1 54, characterized in that the polyethyleneimine has a molecular weight of from about 500,000 to about 1, 400, 000. 1 58. A method as described in claim 141 , characterized in that the vehicle comprises a positively charged polymer that has adhered positively charged branching groups independently selected from - (gly) ni- (arg) n2, H IV-TAT and fragments thereof, PTD Antennapedia and fragments or mixtures thereof, wherein the subscript n 1 is an integer from 0 to about 20, and the subscript n 2 is independently an integer non of from about 5 to about 25. 1 59. A method as described in the reinvidication 158, characterized in that branching groups loaded in positive form are independently selected from groups having the formula - (giy) n? - (arg) n2. 160. A method as described in claim 1 59, characterized in that the subscript n 1 is an integer of from about 1 to about 8. 161. A method as described in the claim 159, characterized in that the subscript n 1 is an integer of from about 2 to about 5. 162. A method as described in the claim 159, characterized in that the subscript n2 is a non-number from from about 7 to about 17. 163. A method as described in claim 159, characterized in that the subscript n2 is a non-number of from about 7 to about 13. 164. A method as described in claim 158, characterized in that the branching groups are selected from HIV-TAT and fragments thereof. 165. A method as described in claim 164, characterized in that branching groups positively loaded with HIV-TAT fragments having the formula (gly) p-RGRDDRRQRRR- (gly) q, (gly) p- YGRKKRRQRRR- (gly) q, or (gly) p-RKKRRQRRR- (gly) q, wherein the subscripts p and q are each independently an integer from 0 to 20. 166. A method as described in the claim 158, characterized in that the branching groups are Antennapedia PTD groups. 167. A method as described in claim 158, characterized in that the positively charged polymer comprises a polypeptide. 168. A method as described in claim 167, characterized in that the polypeptide is selected from polylysines, polyarginines and polyornithines. 169. A method as described in claim 168, characterized in that the polypeptide is a polylysine. 170. A method as described in claim 158, characterized in that the polymer comprises a polymer without peptidyl positively charged. 171. A method as described in claim 170, characterized in that the polymer without peptidyl comprises a positively charged polyalkyleneimine. 172. A method as described in claim 171, characterized in that the polyalkylene imine is polyethylene imine. 173. A method as described in claim 141, characterized in that the composition is a controlled release composition. 174. A method as described in claim 141, characterized in that the antigen suitable for immunization is a botulinum toxin. 175. A method as described in the claim 174, characterized in that the botulinum toxin is a botulinum toxin selected from serotypes A, B, C, D, E, F and G. 176. A method as described in claim 174, characterized in that the botulinum toxin comprises a derivative of botulinum toxin. 177. A method as described in claim 174, characterized in that the botulinum toxin comprises a recombinant botulinum toxin. 178. A method as described in claim 141, characterized in that the antigen is suitable for childhood immunizations. 179. A method as described in claim 141, characterized in that the antigen suitable for immunization is administered to provide resistance to an environmental antigen. 180. A method as described in the claim 141, characterized in that the antigen suitable for immunization is administered to provide resistance to a potential pathogen. 181. A method as described in claim 141, characterized in that the antigen suitable for immunization is administered to provide resistance to a potential biological hazard. 182. A composition as described in claim 4, characterized in that the biologically active agent is an antifungal agent. 183. A composition as described in claim 182, characterized in that it contains from about 1 x 10-10 to about 49.9% by weight of the biologically active agent and from about 1 x 10-9 to about 50% by weight of the vehicle loaded positively. 184. A controlled release composition as described in claim 182. 185. A composition as described in claim 182, characterized in that the anti-fungal agent is selected from amphotericin B, fluconazole, flucytosine, itraconazole, ketoconazole, clotrimazole, econozole, griseofulvin, miconazole, nystatin and cyclopirox. 186. A kit for administering a subject an antifungal agent, characterized in that it comprises an apparatus for delivering the anti-fungal agent to the skin or epithelium of the subject, and a composition as described in claim 1 82. 1 87. A equipment as described in claim 186, characterized in that it also comprises a customary applicator. 1 88. A team as described in the claim 86, characterized in that the composition is contained in an apparatus for administering an anti-fungal agent to a subject through the nail plate or adjacent anatomical structures. 1 89. A device as described in claim 1 86, characterized in that the apparatus is a prosthesis nail plate or lacquer. 190. A method as described in claim 81, characterized in that the biologically active agent is an anti-fungal agent. 1 91. A method as described in the claim 90, characterized in that an anti-fungal agent and vehicle are administered to a subject in a composition containing both components. 1 92. A method as described in claim 1 90, characterized in that the antifungal agent and the vehicle are administer the subject separately. 93. A method as described in claim 1 90, characterized in that the composition is a controlled release composition. 1 94. A method as described in the claim 90, characterized in that the anti-fungal agent is selective for amphotericin B, fluconazole, flucytosine, itraconazole, ketoconazole, clotrimazole, econozole, griseofulvin, miconazole, nystatin and cyclopirox. 1 95. A method as described in the claim 190, characterized in that the anti-fungal agent is administered to treat the symptoms and signs of a fungal infection. 196. A method as described in claim 1 90, characterized in that the anti-fungal agent is administered to alter symptoms or signs of fungal infection of the nail plate or nail bed. 1 97. A peptidyl-free polymer or positively charged polypeptide having branched positively charged branching groups selected independently from - (gly) n? - (arg) n2, HIV-TAT and fragments thereof, and PTD Antennapedia and fragments and mixtures thereof, wherein the subscript n 1 is an integer from 0 to 20, and the subscript n 2 is independently an integer non from about 5 to about 25. 1 98. A polymer without peptidyl or charged polypeptide fit positive as described in claim 1 97, characterized in that branching groups loaded in positive form are independently selected from groups having the formula - (gly) n? - (arg) n2. 99. A polymer without peptidyl or polypeptide positively charged as described in claim 1 98, characterized in that the subscript n 1 is an integer of from about 1 to about 8. 200. A polymer without peptidyl or charged polypeptide in positive form as described in claim 1 98, characterized in that the subscript n 1 is an integer of from about 2 to about 5. 201. A peptidyl-free polymer or polypeptide positively charged as described in claim 1 98, characterized in that the subscript n2 is a non-number of from about 7 to about 17. 202. A polymer without peptidyl or polypeptide positively charged as described in claim 198, characterized in that the subscript n2 is a non-number of from about 7 to about 1 3. 203. A polymer without peptidyl or polypeptide positively charged as described in claim 1 97 , characterized in that the branching groups are selected from HIV-TAT and fragments thereof. 204. A polymer without peptidyl or polypeptide loaded in the form positive as described in claim 203, characterized in that the branched groups loaded in positively adhered form are H IV-TAT fragments having the formula (gly) p-RGRDDRRQRRR- (gly) q, (gly) p-YGRKKRRQRRRq - or (gly) p-RKKRRQRRR- (gly) q, wherein the subscripts p and q are each independently an integer from 0 to 20. 205. A polymer without peptidyl or polypeptide positively charged as described in claim 1 97, characterized in that the branching groups are Antennapedia PTD groups or fragments thereof. 206. A positively charged polymer as described in claim 1 97, characterized in that the positively charged vehicle comprises a polypeptide. 207. A positively charged polymer as described in claim 206, characterized in that the polypeptides are selected polylysines, polyarginines, polyornithines and polyhomoarginines. 208. A positively charged polymer as described in claim 207, characterized in that the polypeptide is a polylysine. 209. A positively charged polymer as described in claim 1 97, characterized in that the positively charged vehicle comprises a polymer without peptidyl positively charged. 21 0. A positively charged polymer as it is described in claim 209, characterized in that the polymer without peptidyl comprises a positively charged polyalkyleneimine. 21 1. A positively charged polymer as described in claim 210, characterized in that the polyalkyleneimine is a polyethyleneimine. 212. A composition comprising a non-covalent complex of: a) a skeleton positively charged; and b) at least two members selected from a group consisting of: i) a negatively charged skeleton having a plurality of adhered image generation portions, or alternatively, a plurality of negatively charged image generation portions; . ii) a negatively charged skeleton having a plurality of adhered steering agents, or alternatively, a plurality of negatively charged steering portions. iii) at least one selected member of RNA, DNA, ribozymes, modified oligonucleotide and cDNA encoding the selected transgene; iv) DNA encoding at least one persistence factor; and v) a negatively charged skeleton having a plurality of attached biological agents, or alternatively, a biological agent loaded in negative form, where the complex carries a positive net charge and at least one of the members is selected from i), ii), iii) or v). 3. A method for preparing a pharmaceutical or cosmetological composition, characterized in that the method comprises combining a positively charged skeletal component and at least 2 members selected from a group consisting of: i) a skeleton loaded in a negative form having a plurality of adhered image generation portions, or alternatively, a plurality of negatively charged image generation portions. ii) a negatively charged skeleton having a plurality of adhered steering agents, or alternatively, a plurality of negatively charged steering portions. iii) at least one selected member of RNA, DNA, ribozymes, modified oligonucleotide and cDNA encoding the selected transgene; iv) DNA encoding at least one persistence factor; and v) a skeleton loaded in a negative form having a plurality of biological agents or cosmetological agents adhered, or a biological agent or cosmetological agent negatively charged; with a pharmaceutically or cosmetologically acceptable vehicle to form a non-covalent complex having a net charge positive, and at least one of the members is selected from i), ii), iii) or v). 214. A composition comprising insulin, and an amount effective for the transdermal delivery of insulin, of a vehicle comprising a positively charged skeleton and having positively charged branching groups adhered thereon, wherein the association between the vehicle and Insulin is non-covalent. 5. A composition as described in claim 214, characterized in that it contains insulin and a positively charged vehicle in a weight ratio of from about 30: 1 to about 1.01: 1. 6. A controlled release composition as described in claim 214. 217. A device for administering insulin to a subject, characterized in that it comprises insulin and a vehicle comprising a positively charged backbone having attached groups of insulin. Branches loaded in positive form and which is in an effective amount for transdermal delivery, where the association between the vehicle and insulin is non-covalent. 8. A device as described in claim 21, characterized in that the composition is contained in an apparatus for administering insulin to a subject through the skin or epithelium. 219. A method for administering insulin to a subject, characterized in that it comprises applying topically to the skin or epithelium of the subject insulin, together with an effective amount of a positively charged vehicle comprising a positively charged backbone having positively charged branching groups adhered therein, wherein Association between the vehicle and insulin is non-covalent. 220. A method as described in claim 21 9, characterized in that the composition is a controlled release composition. 221. A composition comprising an image generating portion and a steering agent and a vehicle comprising a positively charged skeleton having positively charged branch groups adhered thereto and which is in an effective amount for transdermal delivery, in where the association between the vehicle and the biologically active protein is non-covalent. 222. A composition as described in claim 221, characterized in that the image generating portion and the steering agent are physically or chemically distinct. 223. A composition as described in claim 221, characterized in that the image generating portion and the targeting agent are both not phosphate. 224. A composition as described in claim 221, characterized in that the image generating agent is an optical image generation agent. 225. A composition as described in claim 224, characterized in that the image generation agent is selected from Cy3, Cy3.5, Cy5, Cy5.5, Cy7, Cy7.5, Oregon green 488, Oregon green 500, green Oregon 514, green fluorescent protein, 6-FAM, red Texas, HEX, TET and HAMRA. 226. A composition as described in claim 221, characterized in that the image generation agent is suitable for magnetic resonance image generation. 227. A composition as described in claim 221, characterized in that the targeting agent recognizes melanoma. 228. A device for administering to a subject a composition as described in claim 221, characterized in that it comprises an apparatus for supplying the image generating and steering portions and a vehicle comprising a positively charged skeleton having attached positively charged branching groups, and which is in an effective amount for transdermal delivery. 229. A method for administering to a subject an image generation portion and a targeting agent, characterized in that it comprises applying topically to the skin or epithelium of the subject the image generation portion and the targeting agent together with an amount effective of a positively charged vehicle comprising a positively charged skeleton that it has positively charged branching groups adhered to, where the association between the vehicle and the biologically active protein is non-covalent. 230. The method as described in claim 229, characterized in that the image generating portion and the steering agent are physically or chemically distinct. 231. The method as described in claim 229, characterized in that the image generation portion and the targeting agent are both not phosphate. 232. The method as described in the claim 229, characterized in that the image generation agent is an optical image generation agent. 233. The method as described in claim 232, characterized in that the image generating agent is selected from Cy3, Cy3.
3. 3, Cy5, Cy5.5, Cy7, Cy7.5, Oregon green 488, Oregon green 500 , Oregon green 514, green fluorescent protein, 6-FAM, Texas red, Hex, TET and HAMRA. 234. The method as described in claim 229, characterized in that the image generating agent is suitable for magnetic resonance image generation. 235. The method as described in claim 229, characterized in that the targeting agent recognizes melanoma. 236. The method as described in claim 229, characterized in that the composition is applied to classify Patients at risk of melanoma. 237. The method as described in claim 229, characterized in that the composition is applied to aid in the surgical removal of melanoma. 238. The method as described in the claim 229, characterized in that the composition is applied together with photographic techniques or image analysis techniques. 239. A composition comprising a non-covalent complex of: a) a skeleton positively charged; and b) at least two members selected from a group consisting of: i) a negatively charged skeleton having a plurality of adhered image generating portions; or a plurality of negatively charged image generation portions; ii) a negatively charged skeleton having a plurality of adhered steering agents; or a plurality of negatively charged address portions; and iii) a negatively charged backbone having a plurality of attached biological agents, or a biological agent negatively charged; where the complex carries a positive net charge and at least one of the members is selected from i), ii), iii) or v). 240. A method for preparing a pharmaceutical composition or Cosmetology, characterized in that the method comprises combining a positively charged skeletal component and at least two members selected from the group consisting of: i) a negatively charged skeleton having a plurality of adhered image generating portions or as an alternative , a plurality of negatively charged image generation portions; ii) a negatively charged skeleton, having a plurality of adhered steering agents, or alternatively, a plurality of negatively charged steering portions; and iii) a skeleton loaded in a negative form having a plurality of biological agents or cosmetological agents adhered, or a biological agent or cosmetological agent negatively charged with a pharmaceutically or cosmetologically acceptable carrier to form a non-covalent complex having a net charge positive, provided that at least one of the members is selected from i), ii), iii) or v). R E S U M E Compositions and methods are provided that are useful for the delivery, including transdermal delivery, of biologically active agents, such as non-protein, non-nucleotide therapeutics and protein-based therapeutics excluding insulin, botulinum toxins, antibody fragments and VEGF. The compositions and methods are particularly useful for the topical delivery of antifungal agents and antigenic agents suitable for immunization. Alternatively, the compositions may be prepared with components useful for directing the delivery of the compositions, as well as the imaging components.