CA2543370A1

CA2543370A1 - Formulations for topical delivery of bioactive substances and methods for their use

Info

Publication number: CA2543370A1
Application number: CA002543370A
Authority: CA
Inventors: Vromen Jacob
Original assignee: Individual
Current assignee: Australian Importers Ltd
Priority date: 2002-10-25
Filing date: 2003-10-15
Publication date: 2004-05-13
Also published as: US20070071711A1; US7241456B2; US20040081681A1; WO2004039348A1; AU2010201779A1; AU2003282834A1; US9162084B2; EP1558206A1

Abstract

The invention relates to topical delivery of bioactive agents. More particularly, the invention relates to anhydrous formulations for percutaneo us absorption. The invention provides formulations that allow efficient topical delivery of high concentrations of bioactive substances for percutaneous absorption. The formulations according to the invention are generally non- irritating to the skin. The formulations comprise an exfolient and a bioacti ve substance in micronized form.

Description

FORMULATIONS FOR TOPICAL DELIVERY OF BIOACTIVE SUBSTANCES
AND METHODSFOR THEIR USE
BACKGROUND OF THE INVENTION
Field of the invention The invention relates to topical delivery of bioactive agents. More particularly, the invention relates to anhydrous formulations for percutaneous absorption.
Summary of the related art Topical administration of biologically active agents has become an important method for treating a variety of skin conditions. Carlin, Cosmetic Dermatology, February 2001, pp. 35-38 teaches topical administration of vitamin C to reduce erythema of acne rosacea. Greco, Plastic and Reconstructive Surgery 105: 464-465 (2000) suggests the use of topical vitamin C in the treatment of fine wrinkles and as a stimulant for wound healing. Norman and Nelson, Skin and Aging, February 2000, pp. 28-33 teaches topical administration of a variety of common herbs to treat various dermatologic conditions.
Unfortunately, many bioactive substances are not efficiently absorbed percutaneously. To overcome this problem, scientists have utilized low pH formulations or derivatives of bioactive substances. However, low pH
formulations are irritating to the skin and derivatized~
compounds, while they may be more efficiently absorbed, are not generally efficiently bioconverted to yield the active compound. In addition, it is frequently difficult to obtain high concentrations of active in topical formulations, and in aqueous formulations, hydrophobic actives cannot be used.
There is, therefore, a need for new formulations that allow efficient topical delivery of high concentrations of underivitized bioactive substances for percutaneous absorbtion. Ideally, such formulations should be non-irritating to the skin.

BRIEF StIN~lARY OF THE INVENTION
The invention provides formulations that allow efficient topical delivery of high concentrations of bioactive substances for percutaneous absorption. The formulations according to the invention are generally non-irritating to the skin.
In a first aspect, the invention provides formulations that are capable of efficient percutaneous absorption of high concentrations of hydrophobic, hydrophilic or ampiphilic bioactive substances. A great variety of bioactive substances may be included in the formulations according to the invention.
The formulations according to the invention comprise an anhydrous carrier medium, a high concentration of bioactive substance, and an exfoliant. Such formulations are free of any occlusive agent that prevents percutaneous absorption.
In a second aspect, the invention provides methods for using the formulations according to the invention to treat a dermatologic condition, the methods comprising applying therapeutically effective amounts of the formulations according to the invention to the skin.
In a third aspect, the invention provides a method for treating a dermatologic condition, the method comprising exfoliating the skin and applying to the skin a formulation comprising an anhydrous carrier medium and a high concentration of bioactive substance. The method may optionally include in the formulation an exfoliant.

In a fourth aspect the invention provides a method for introducing a bioactive substance into the bloodstream of a patient. The method according to this aspect of the invention comprises applying therapeutically effective amounts of the formulations according to the invention to the skin.
In a fifth aspect the invention provides a method for introducing a bioactive substance into the bloodstream of a patient. The method according to this aspect of the invention comprises exfoliating the skin of a patient and applying to the skin a formulation comprising an anhydrous carrier medium and a high concentration of bioactive substance.

DETAINED DESCRIPTION OF THE PREFERRED EI~ODIL~NTS
The invention relates to topical delivery of bioactive agents. More particularly, the invention relates to anhydrous formulations for percutaneous 5 absorption. The patents and publications cited herein reflect the knowledge in the art and are hereby incorporated by reference in their entirety. Any inconsistency between these patents and publications and this specification shall be resolved in favor of the latter .
The invention provides formulations that allow efficient topical delivery of high concentrations of bioactive substances for percutaneous absorption. The formulations according to the invention are generally IS non-irritating to the skin, in spite of high concentration, which may cause slight tingling of a passive nature due to the heightened activity.
The formulations according to the invention provide many advantages over aqueous formulations. For example, acidic or basic biological actives in an aqueous environment will at high concentrations affect the pH of the formulation, thus rendering it irritating to the skin. In an anhydrous environment, ionization does not occur, so high concentrations of such actives may be achieved in a formulation that does not irritate the skin. This is advantageous because one of the governing factors for percutaneous absorption is the concentration of the biological active, with higher concentrations leading to increased percutaneous absorption.

Some embodiments of the formulations according to the invention provide the further advantage of providing the biological active in a particle size that approximates the molecular size of the biological active.
This is advantageous because the size of the biological active is another governing factor for percutaneous absorption. As particle size decreases, percutaneous absorption increases.
Another advantage provided by the anhydrous formulations according to the invention is increased stability of the biological active. Hydrophilic biological actives are frequently labile when exposed to water. They are prone to oxidation, hydrolysation and decomposition. In the formulations according to the invention, they are not. Also, in some embodiments of the invention, this stability is further enhanced by coating the molecular-sized particles with protective oils. This allows non-derivatized (i.e., not covalently modified) biological actives to be used and prevents coalescence of particles. Since bioconversion of covalently derivatized biological actives is generally inefficient, this provides greater activity for the biological active.
Yet another advantage provided by the anhydrous formulations according to the invention is that they efficiently partition hydrophilic biological actives for percutaneous absorption, because the hydrophilic molecules prefer the hydrophilic environment of the dermis to the hydrophobic environment of the anhydrous formulation.
An additional advantage provided by the formulations according to the invention is that they exfoliate the skin as they are applied. By removing dead skin cells of the stratum corneum without damaging underlying keratinocytes and fibroblasts, percutaneous absorption is further enhanced. In some embodiments of the invention, this effect is increased through the use of penetration enhancers that reversibly alter the physiochemical nature of the stratum corneum to reduce its diffusional resistance.
In a first aspect, the invention provides formulations that are capable of efficient percutaneous absorption of high concentrations of hydrophobic, hydrophilic or amphoteric bioactive substances. A great variety of bioactive substances may be included in the formulations according to the invention. For purposes of the invention, "percutaneous absorption" means penetration of the stratum corneum by the bioactive substance in active form. "Active form" means that the bioactive substance is capable of performing its physiological role.
The formulations according to the invention comprise an anhydrous carrier medium, a high concentration of micronized bioactive substance, and an exfolient. In certain embodiments, such formulations further comprise a penetration enhancer. Such formulations are preferably free of any occlusive agent that prevents percutaneous absorption, such as silicones. In preferred embodiments the micronization may be wet micronization or dry micronization. For purposes of the invention, an "anhydrous carrier medium" is a substance that is substantially free of water. "Substantially free of water" means that water that is present, if any, is in insufficient amount to affect either percutaneous absorption or the activity or stability of the bioactive substance. Preferred anhydrous carriers include, without limitation, esters, amides, ethoxylated fats, mineral oil, petrolatum, vegetable oils, animal fats, triglycerides, polyols (e. g., glycerol), glycerin, propylene glycol and sorbitol. Glycerin in concentrationsrof 20-400, 25-35o and about 35o are among the preferred embodiments. In certain preferred embodiments the exfolient is non-irritating. In certain preferred embodiments the bioactive substance is non-derivitized.
A "high concentration" is a concentration that is therapeutically effective, as defined for the second aspect of the invention. In many cases, such concentration exceeds the concentration of the bioactive substance obtainable in an aqueous medium under conditions that are not irritating to the skin. "Non-derivativitized" means that the bioactive substance is in its native form and has not been covalently modified, e.g., through esterification, etc.
In certain embodiments, the formulations may be manufactured by standard "dry" micronization procecess.

According to certain preferred methods of manufacturing the preferred formulations, the bioactive substance, preferably in powder form is subjected to a "wet"
micronization process, as made available by Microniser Pty. Ltd. of Dandenong, Australia / Micronisers of Australia of Melbourne, Australia. Wet micronization prevents overheating of the active, the coating prevents coalescence of particles and protects against oxidation, reduction and hydrolysis. This process, which may be contrasted to so-called "dry" or standard micronization processes, preferably involves the grinding of the powder, suspended in or otherwise in the presence of a non-aqueous liquid, preferably an oil (hereinafter, the "suspending medium"). The process is preferably conducted in an abrasion-resistant container in the presence of a grinding medium, using sufficiently high rpm for a sufficiently long duration, and a suitable stirrer. The resulting suspension may be separated from the grinding medium by suction filtration of the powder.
This micronization process is capable of producing particles of bioactive substance having a mean particle size corresponding to the molecule size of the bioactive substance.
Alternatively, the grinding may be conducted in the presence of 0.1 to 300, and preferably 0.5 to 15~ by weight, of a grinding aid such as an alkylated vinylpyrrolidone polymer, a vinylpyrrolidone-vinylacetate copolymer, an acylglutamate, an acrylate-tert.-octylpropenamide copolymer, a ditolylether sulphonic acid-formaldehyde condensate, a carbomer, a commercial mixture of fatty acid esters comprising a nonionic precursor such as tristyrylphenol ethoxylate or, in particular, a phospholipid, as described in U.S. Patent 5,869,030.
5 The suspending medium is most preferably a vegetable oil, which promotes (along with the physical micronization process, as described above) breaking the bioactive substance into ultrafine particles and, at the same time, coating the particles with the oil, which 10 promotes maximum absorbance and stability of the bioactive substance in the formulation. The micronized bioactive substance particles used preferably exhibit a mean particle size of no more than approximately 5 Lun, and preferably a mean particle size of in the range of IS from about 0.01 to about 2 Vim, and most preferably from about 0.05 to about 1.5 Vim, and especially from about 0.1 to about 1Ø Eun.
Oils most preferable and therefore most suitable for use include, without limitation, caprylic triglycerides, capric triglycerides, isostearic triglycerides, adipic triglycerides, propylene glycol myristyl acetate, lanolin oil, polybutene, isopropyl palmitate, isopropyl myristate, diethyl sebacate, diisopropyl adipate, hexadecyl stearate, cetyl oleate, oleyl alcohol, hexadecyl alcohol, wheatgerm oil, vegetable oils such as castor oil, corn oil, cottonseed oil, olive oil, palm oil, coconut oil, palm kernel oil, canola oil, sunflower oil, safflower oil, meadow foam oil, jojoba oil, hydrogenated vegetable oils, and mineral oil. In one preferred embodiment, the bioactive substance is micronized in the presence of capric/caprylic glycerides in which the bioactive substance is present at high concentrations.
A "bioactive substance" (sometimes referred to as a "biological active") is a substance that exerts some physical effect upon the skin. Such bioactive substances may provide protective functions, such as protection from exposure from radiation (e. g., from sunlight) or chemical exposure. The bioactive substance may serve a therapeutic role for the treatment of a dermatological condition, or may provide a micronutrient. Many bioactive substances are discussed herein. However, those skilled in the art will recognize that other well known bioactive substance, or any bioactive substance developed in the future can readily be incorporated into the topical delivery formulations according to the invention. The skilled artisan will also recognize that combinations of different bioactive substances can readily be incorporated into the topical delivery formulations according to the invention. -In certain embodiments the bioactive substance is a vitamin or a micronutrient. Preferred vitamins or micronutrients include, without limitation, vitamin C, vitamin E, vitamin B12, vitamin B6, folic acid, beta carotene and niacin or combinations thereof. A
combination of vitamin C and vitamin E is one preferred embodiment. This preferred embodiment may further include niacin. Niacin in combination with an anti-inflammatory agent is another preferred embodiment.
Preferred anti-inflammatory agents include, without limitation, aspirin, ibuprofen and acetaminophen and cox-2 inhibitors.
In certain embodiments the bioactive substance is an antimicrobial or antiviral antibiotic. Preferred antimicrobial antibiotics include antibacterial antibiotics antiparasitic antibiotics and antifungal antibiotics. Preferred fungicides include, without limitation, butocouazole nitrate, haloprogen, clotrimazole and other azoles. Preferred antivirals include, without limitation, 0-[(2-hydroxyetoxy)-methyl]guanine and other herpes treatment medications, and tee tree oil (oil of Melaleuca spp.).
In certain embodiments the bioactive substance is an antioxidant selected from, without limitation, urocanic acid and other imidazoles; D,L-carnosine, D-carnosine, L-carnosine, anscrine and other peptides; alpha-carotine, beta-carotine, lycopine and other carotines; carotenoids;
dihydrolipoic acid and other lipoic acids;
aurothioglucose, propylthiouracil, thioredoxin, glutathion, cysteine, cystine, cystamine and other thiols; dilauryl thioproponate; distearyl thiopropionate;
thiopropionate; thiopropionic acid; butathione-sulfoxamines, homocysteine-sulphoxamine, butathione-sulphones, penta-, hexa- and heptathioninesulphomimine and other sulfoxamine compounds; alpha-hydroxy-fatty acids, palmitic acid, lactoferrin, EDTA, EGTA and other metal chelating agents; citric acid, lactic acid, malic acid and other alpha-hydroxy acids; gamma-linolenic acid, linoleic acid, oleic acid and other unsaturated fatty acids; folic acid; ubiquinone, ubiqinol and other quinones; vitamin C; ascorbyl palmitate, Mg ascorbyl phosphate, ascorbyl acetate and other vitamin C
derivatives; vitamin E acetate and other tocopherols and tocopherol derivatives; retinoids; vitamin A; vitamin A
palmitate and other vitamin A derivatives; coniferyl benzooate of benzoin resin; rutic acid; alpha-glycosylrutin; ferulic acid furfurylideneglucitol;
carnosine; butylhydroxytoluene (BHT); butylhydroxyanisole (BHA); nordihydroguaic resin acid; nordihydroguaiaretic acid; trihydroxybuterophenone; uric acid; mannose; ZnO, Zn04 and other zinc compounds; selenium and stilbenes.
In certain embodiments the bioactive substance is a sugar, an amino acid or a small peptide, including, without limitation carnosine, acetyl-L-carnithine, N-acetyl-carnithine, N-acetyl-cysteine, N-acetyl-D-glucosamine, B/D/L-alanine, alpha-keto-glutarates, arginates, L-arganine base/HCl, L-arginine-pyroglutamate, ascorbates, L-aspargine monohydrate, aspartame, aspartates, L-aspartic acid, L-carnithine base, L-carnithine fumarate, L-carnithine/HCl, L carnithine bitartrate, L-carnithine-L-tartrate, chelates, L-citruline, creatin monohydrate, creatin phosphate, creatine pyruvate, L-cysteine base, L-cysteine/HC1 monohydrate, anhydrous L-cysteine/HCl, cysteinates, N,N-dimethylglycine base/HCl, glutamates, L-glutamic acid, L-glutamine, L-glutamine peptide, reduced L-glutathione, L-glycine, keto-glutaric acid, L-histidine base/HC1, L-isoleucine, L-leucine, lysinates, L-lysine monohydrate/HC1, D-mannose, DL/L-methionine, L-ornithine/HCl, DL/L-phenylalanine, L-hydroxyproline, trans-hydroxy-L-proline, pyroglutamic acid, D/L-ribose, L-selenium-methionine, L-serine, taurates, tartarates, L-threonine, trimethylglycine, DL/L-tryptophan, taurine, L-theanine, L-tyrosine, L-valine, xylitol, D-xylose,DL/L-zinc monomethionine, and all other D or L-amino acids and short peptides and all bases, acids and salts thereof.
In certain embodiments the antioxidant may be an antioxidant derivative. Preferred derivatives include, without limitation, esters, ethers, peptides, lipids, nucleotides and nucleosides of such antioxidants.
Preferred derivatives also include glycosyl, N-acetyl, methyl, ethyl, propyl, amyl, butyl lauryl, palmitoyl, oleyl, gamma-linoloyl " cholesteryl and glyceryl esters of such antioxidants.
In certain embodiments the bioactive substance is a biological additive. As used herein, the term "biological additive" indicates any compound obtained from a natural source, including plants, animals, bacteria and yeast, which has a medicinal or otherwise beneficial effect when topically applied to the skin.
Examples of biological additives include, without limitation, oil of Melaleuca spp. (tea tree oil), oil of Lavandula angustifolia, Carica papaya extract, Echinacea angustifolia extract, Mimosa tenuiflora extract, Hydrocotyl (centella) asiatica extract, gingko biloba extract, Matricaria chamomila (chamomile oil) extract, Hypericum perforatum extract, Aloe barbedensis extract, and the like. The biological sources for "biological additive" may also include, but are not limited to the following: Aloe Vera, (e. g., Aloe Barbedensis); Arnica, (e.g.,Arnica Montana); Bladderwrack (seaweed), 5 (e.g.,Fucus Vesciculosis); Birch, (e.g.,Betula Alba) (Pendula); Chamomile, (e.g.,Matricaria Chamomila, Chamomila Recutita); Marsh Mallow, (e.g.,Althea Officinalis); Meadow Sweet, (e.g.,Spirea Ulmaria) (Filipendula); Mint/Lemon Balm, (e.g.,Melissa 10 Officinalis); Mimosa, (e.g.,Mimosa Tenuiflora); Myrrh Tincture, (e.g.,Commiphor Myrrha); Neem, (e.g.,Melia Azadirachta); Nettle (stinging), (e.g.,Urtica Dioica);
Papaya, (e.g.,Carica Papaya); Propolis (bee glue), (e.g.,Propolis Cera); Raspberry, (e.g.,Rubis Idaeus); Red 15 Poppy, (e.g.,Papaver Rhoeas); Rose Hip (dog rose), (e.g.,Rosa Carima); Rosemary, (e.g.,Rosemarinus Officinalis); Sage, (e.g.,Salvia Officinalis); St. Johns Wort, (e.g.,Hypericum Perforatum); Strawberry, (e.g.,Fragaria Vesca); Thea Sinensis (green tea), (e.g.,Camelia Sinensis); Walnut, (e.g.,Juglans Regia);
Witchhazel (dist/extr), (e.g.,Hamamelis Virginiana);
Yarrow, (e.g.,Achillea Millefolium); Wild Yam, (e.g.,Dioscorea Villosa); Hawthorn, (e.g.,Crataegus Monogina/Oxyantha); Herma (black/rod), (e.g.,Lawsoma Ehemus); Hops, (e.g.,Humulus Lupulus); Horse Chestnut, (e.g.,Aesculus Hippocastanum); Horse Tail, (e.g.,Equisitum Arvense); Ivy, (e.g.,Hedera Helix);
Linden/Lime Tree Blossoms, (e.g.,Tilia Argentea Cordata);
Madder, (e.g.,Rubia Tinctorum); Marigold, (e.g.,Calendula Officinalis, Centella Asiatica, Centella Asiatica Urban, Hydrocotyl Asiatica); Carrot (roots), (e.g.,Daucus Carota); Comfrey (Allantoine), (e.g.,Symphytum Officinale); Coneflower (Echinacea), propolis(e.g.,Echinacea Angustifolia); Cucumber, (e.g.,Cucumis Sativus, Frucus Cucumis); Fenugreek, (e.g.,Trigonella Foenum Greacum); Gingko, (e.g.,Gingko Biloba); Ginseng, (e.g.,Panax Ginseng); Great Burdock, (e.g.,Radix Bardanea/Arctium Zappa); Tea Tree Oil, (e.g.,0i1 of Melaleuca Alternifolia); Colts Foot, (e.g.,Tussilago Farfara); Clover, arbutui (e.g.,Trifolium Pratense); Speedwell, (e.g.,Veronica Officinalis). A
particularly preferred biological additive is tea tree oil. In a preferred embodiment, one or more biological additive is present in the formulation in a combined amount of from about 1% to 10% by weight, more preferably from about 2% to 8% by weight, and most preferably from about 4% to 6% by weight.
In certain embodiments the bioactive substance is a sunblock. In certain embodiments the sunblock is not a UV absorber. Preferred sunblocks include, without limitation, dioxybenzone, ethyl 4-[bis(hydroxypropyl)]
aminobenzoate, glyceryl aminobenzoate, homosalate, menthyl anthranilate, octocrylene, octyl methoxycinnamate, octyl salicylate, oxybenzone, padimate O, red petrolatum, titanium dioxide, 4-methylbezylidene camphor, benzophenone-1, benzophenone-2, benzophenone-6, benzophenone-12, isopropyl dibenzoyl methane, butylmethoxydibenzoylmethane, zotocrylene, zinc oxide, para-aminobenzoic acid, cinnamate; and derivatives, analogs and functional analogs of such sunblocks.

In certain embodiments the bioactive substance is a local anesthetic. Preferred local anesthetics include, without limitation, lidocaine and procaine. See also Martindale, The Extra Pharmacopia, Twenty-eighth Edition, The Pharmaceutical Press, London (1982), pp. 899-923.
For purposes of the invention, an "exfoliant" is a substance that eliminates dead cells from the stratum corneum without killing underlying living skin cells. In certain embodiments the exfoliants are chemical exfoliants, such as AHA and BHA. In certain preferred embodiments, the exfoliating agents are enzymes. One preferred enzyme is papain, an enzyme obtained from unripe papaya. One particularly preferred form of papain is Linked-PapainTM (papain carbomer, as described in CTFA, the International Cosmetic Ingredients Dictionary) in which papain is covalently immobilized to 1a polyacrylic acid (900,000 daltons), commercially available from Collaborative Laboratories, 3 Technology Drive, East Setauket, NY 11733). In a preferred embodiment, the enzyme is present in the formulation in an amount between about 1% and 7~, more preferably between about 2% and 6~, most preferably about 5o by weight. The ability of papain to act as an exfoliant allows enhancement of percutaneous absorption of any desired bioactive substance. Other preferred enzymes include, without limitation, serine proteases, macramidase, penicillinase, pepsin, plasmin, bromelains, streptokinase, sutilains, trypsin, urokinase, keratinase, amylase, hyaluronidase, cholic acid, chymopapain, chymotrypsin, cynara, brinolase and chenodeoxycholic acid. See also Martindale, The Extra Pharmacopia, Twenty-eighth Edition, The Pharmaceutical Press, London (1982), pp. 644-661.
Certain preferred embodiments of the topical delivery formulations according to the invention include penetration enhancers. For purposes of the invention, a "penetration enhancer'° is an agent that increases skin permeability by revaersibly altering the physiochemical nature of the the stratum corneum to reduce its diffusional resistance. Preferred penetration enhancers include, without limitation, oxazolidinones, propylene glycol, epidermal enzymes, oleic acid,dimethyl isosorbide, dimethylsulfoxide, ethanol, diethylene glycol monoethyl ether, hyaluronic acid, chitin, mucopolysaccharides, unsaturated fatty acids, linoleic acid, alpha linoleic acid, cod liver oil, menthol derivatives, squalene, glycerol derivatives, glycerol monoethers; chamomile flavones apigenin, lutrolin, apigenin 7-O-beta-glucoside and other herbal ingredients.
Without wishing to be bound by theory, the inventor believes that such penetration enhancers act through one or more of the following mechanisms: increasing the fluidity of the stratum corneum lipids and reducing the diffusional resistance to permeants; removing intercellular lipids and dialation between adherent cornified cells; increasing the thermodynamic activity of drugs in vehicles; exfoliating stratum corneum cell membranes; dissociating adherent cornified cells and elimination of the barrier function.

In a second aspect, the invention provides methods for using the formulations according to the invention to treat a dermatologic condition, the methods comprising applying the formulations according to the invention to the skin. The formulations are applied at a therapeutically effective concentration. A
"therapeutically effective concentration" is a concentration that eliminates or reduces recognizable clinical manifestations of a dermatological condition. A
"dermatological condition" is an abnormality of the skin.
Dermatological conditions include, without limitation, acne, bruises, burns, eczema, mycoses, pruritis, psoriasis, seborrhea, scabs, shingles, tineapedis, wounds, wrinkles and erythema of acne rosacea. Preferred agents for treating dermatological conditions include, without limitation, anti-acne preparations; anti-inflammatory agents; monobenzone and other depigmenting agents;
amcinonide, diflorasone diacetate, hydrocortisone and other dermatitis relief agents; methylbenzethonium chloride and other diaper rash relief agents; mineral oil, PEG-4 dilaurate, lanolin oil, petrolatum, mineral wax and other emolients and moisturizers; alclometasone dipropionate, betamethasone valerate, isopropyl myristate MSD and other pruritic medications; anthralin, methoxsalen, coal tar and otherpsoriasis, seborrhea and scabicide agents; 2-(acetyloxy)-9-fluoro-1',2'3'4'-tetrahydro-11-hydroxypregna-1,4-dieno[16,17b] napthalene-3,20-dione, 21-chloro-9-fluoro-1',2',3',4'-terahydro-11b-hydroxypregna-1,4-dieno[16z,17b] napthalene-3,20-dione and other steroids.

In certain embodiments, the method according to this aspect of the invention, the method further includes preventing the formulation according to the invention from escaping to the exterior of the skin. Preferred 5 methods for achieving this include covering the area of the skin that has been treated with a formulation according to the invention with an occlusive patch or other occlusive agent.
In a third aspect, the invention provides a method 10 for treating a dermatologic condition, the method comprising exfoliating the skin and applying to the skin a formulation comprising an anhydrous carrier medium and a high concentration of a bioactive substance. The method may optionally include in the formulation an 15 exfoliant, preferably a non-irritating exfoliant.
Exfoliation of the skin may be chemical, mechanical or enzymatic. Chemical and enzymatic exfoliation have been discussed previously. Mechanical exfoliation may be carried out by any mechanical frictional force, 20 including, without limitation, brushing, washing or particle pressure. In certain preferred embodiments, the formulation contains a wet-micronized bioactive substance. The term "dermatologic condition" is as described previously.
In certain embodiments, the method according to this aspect of the invention, the method further includes preventing the formulation according to the invention from escaping to the exterior of the skin. Preferred methods for achieving this include covering the area of the skin that has been treated with a formulation according to the invention with an occlusive patch or other occlusive agent.
In a fourth aspect the invention provides a method for introducing a bioactive substance into the bloodstream of a patient. The method according to this aspect of the invention comprises applying therapeutically effective amounts of the formulations according to the invention to. the skin.
In certain embodiments, the method according to this aspect of the invention, the method further includes preventing the formulation according to the invention from escaping to the exterior of the, skin. Preferred methods for achieving this include covering the area of the skin that has been treated with a formulation according to the invention with an occlusive patch or other occlusive agent.
In a fifth aspect the invention provides a method for introducing a bioactive substance into the bloodstream of a patient. The method according to this aspect of the invention comprises exfoliating the skin of a patient and applying to the skin a formulation comprising an anhydrous carrier medium and a high concentration of bioactive substance.
In certain embodiments, the method according to this aspect of the invention, the method further includes preventing the formulation according to the invention from escaping to the exterior of the skin. Preferred methods for achieving this include covering the area of the skin that has been treated with a formulation according to the invention with an occlusive patch or other occlusive agent.
The following examples are intended to further illustrate certain particularly preferred embodiments of the invention and are not intended to limit the scope of the invention in any way.

Example 1 Preparation of a formulation for percutaneous absorption Table 1: Preferred concentrations Examples of agents Purpose Conc.

Anhydrous Glycerin Humectant 35%

propylene glycol skin 10%

Medium conditioner capric-caprylic emollient 10%

triglyceride cetearyl alcohol viscosity 4.5%

increasing agent ceteth-20 surfactant -d-tocopherol antioxidant 0.6%

apricot kernel oil skin -conditioner sweet almond oil skin -conditioner ascorbyl palmitate antioxidant 0.6%

thiodiproprionic acid antioxidant 0.6%

BHT antioxidant 0.3%

Phenoxyethanol preservative 0.6%

Methyl/ethyl/propyl/butyl preservatives 0.3%

parabens Strawberry extract fragrance -Examples of agents Purpose Conc.

Bioactive Ex.l: micronized niacin- 4%

nicotinic acid substance (nicotinamide)in capric/caprylic triglyceride 50% acetylsalicylic acid 5%

micronized in capric/caprylic triglyceride 25-30%

micronized L-ascorbic acid in capric/caprylic triglyceride 10%

Ex.2: 50% acetylsalicylic acid micronized in 4-5%

capric/caprylic triglyceride 25-30%

Ex.3: L-carnosine micronized in 8%

capric/caprylic triglyceride 0.5%

Micronized L-ascorbic acid 0.5%

in capric/caprylic 0.5%

triglyceride 0.5%

Ex.4: Extracts in propylene 5%

or butylenes glycol:

Witch hazel Horse chestnut Arnica Chamomile Corn poppy Ex.5: Lidocaine micronized in capric/caprylic triglycerides Exfoliant papain carbomer 5%

Example 2 Application of a formulation for percutaneous absorption Patients were 40-55 years of age with significant wrinkling and photo-damage. Composition for topical 5 administration contained 30o wet micronized (in capric/caprylic triglyceride) Z-ascorbic acid; 2°s alpha-tocopheral, 35o glycerin and 5% papain carbomer. This composition was applied sparingly to the affected area and gently massaged into the skin every other night. The 10 area was moisturized on alternate (non-treatment) nights.
Treatment was repeated over a 90 day course. Substantial improvement of wrinkling and photo-damage was observed.
Example 3 Application of a formulation to 15 obtain blood levels of active Balb/c mice are obtained and shaved in the dorsal area. The composition is 50% acetylsalicylic acid wet micronized in capric/caprylic triglyceride; 35~ glycerin and 5% papain carbomer. The composition is applied 20 sparingly into the shaved area and gently massaged into the skin. One hour later, plasma is obtained and acetylsalicylic acid is measured by the method of Coudray et al., J. Chromatogr. 34: 166-173 (1996). Briefly, aliquots (400 uZ) of serum are acidified with 75 uL
25 concentrated HC1 (350) in 10 x 70 mm glass tubes. The samples are vortexed for 30 seconds, then 3mZ ether is added and the solution is extracted for 2 minutes. The tubes are centrifuged at 1600 x g for 15 minutes, then 2mL of ether phase is removed and dried under a nitrogen steam. The dry residue is reconstituted in 200 uL of mobile phase (85% methanol, sodium citrate and acetate at pH5.45) and loaded into a reversed phase HPLC column (5~Zm ultrasphere octadecyl silane; Alltech, Paris, France).
Flow rate is 1 mL/minute and detection is at 295 nm. It is expected that acetylsalicylic acid will be detected in the sample.

Claims

What is claimed is:

1. A formulation for topical application comprising an anhydrous carrier medium, a high concentration of micronized bioactive substance, and an exfolient.

2. The formulation according to claim 1 further comprising a penetration enhancer.

3. The formulation according to claim 1 which is free of any occlusive agent that prevents percutaneous absorption.

4. The formulation according to claim 1 wherein the anhydrous carrier is selected from the group consisting of esters, amides, ethoxylated fats, mineral oil, petrolatum, vegetable oils, animal fats, triglycerides, polyols, glycerin, propylene glycol and sorbitol.

5. The formulation according to claim 4, wherein the anhydrous carrier is glycerin.

6. The formulation according to claim 5 wherein glycerin is present in concentrations of 20-40%.

7. The formulation according to claim 6 wherein glycerin is present in concentrations of about 35%.

8. The formulation according to claim 1 wherein the exfolient is non-irritating.

9. The formulation according to claim 1 wherein the bioactive substance is non-derivitized.

10. The formulation according to claim 2 wherein the penetration enhancer is selected from the group consisting of oxazolidinones, propylene glycol, epidermal enzymes, oleic acid,dimethyl isosorbide, dimethylsulfoxide, ethanol, diethylene glycol monoethyl ether, hyaluronic acid, chitin, mucopolysaccharides, unsaturated fatty acids, linoleic acid, alpha linoleic acid, cod liver oil, menthol derivatives, squalene, glycerol derivatives, glycerol monoethers; chamomile flavones apigenin, lutrolin, apigenin and 7-O-beta-glucoside.

11. The formulation according to claim 2 wherein the penetration enhancer acts through one or more of the following mechanisms: increasing the fluidity of the stratum corneum lipids and reducing the diffusional resistance to permants; removing intercellular lipids and dialation between adherent cornified cells; increasing the thermodynamic activity of drugs in vehicles;
exfoliating stratum corneum cell membranes;
dissociating adherent cornified cells and elimination of the barrier function.

12. The formulation according to claim 1 made by subjecting the bioactive substance in powder form to a wet micronization process.

13. The formulation according to claim 12 wherein the micronization process comprises grinding the powder, suspended in or otherwise in the presence of a non-aqueous liquid.

14. The formulation according to claim 13, wherein the non-aqueous liquid is an oil.

15. The formulation according to claim 14 wherein the oil is selected from the group consisting of caprylic triglycerides, capric triglycerides, isostearic triglycerides, adipic triglycerides, propylene glycol myristyl acetate, lanolin oil, polybutene, isopropyl palmitate, isopropyl myristate, diethyl sebacate, diisopropyl adipate, hexadecyl stearate, cetyl oleate, oleyl alcohol, hexadecyl alcohol, wheatgerm oil, vegetable oils such as castor oil, corn oil, cottonseed oil, olive oil, palm oil, coconut oil, palm kernel oil, canola oil, sunflower oil, safflower oil, meadow foam oil, jojoba oil, hydrogenated vegetable oils, and mineral oil.

16. The formulation according to claim 15 in which the bioactive substance is micronized in the presence of capric/caprylic glycerides.

17. The formulation according to claim 13 wherein the micronization process produces particles of bioactive substance having a mean particle size corresponding to the molecule size of the bioactive substance.

18. The formulation according to claim 13 wherein the grinding is conducted in the presence of 0.1 to 30%, by weight, of a grinding aid selected from an alkylated vinylpyrrolidone polymer, a vinylpyrrolidone-vinylacetate copolymer, an acylglutamate, an acrylate-tert.-octylpropenamide copolymer, a ditolylether sulphonic acid-formaldehyde condensate, a carbomer, a mixture of fatty acid esters comprising a nonionic precursor, tristyrylphenol ethoxylate and a phospholipid.

19. The formulation according to claim 18 wherein the grinding aid is present in the amount of 0.5 to 15% by weight.

20. The formulation according to claim 12 wherein the micronized bioactive substance particles used exhibit a mean particle size of no more than approximately 5 µn.

21. The formulation according to claim 20, wherein the micronized bioactive substance particles used exhibit a mean particle size of no more than a mean particle size of in the range of from about 0.01 to about 2 µm.

22. The formulation according to claim 21, wherein the micronized bioactive substance particles used exhibit a mean particle size of no more than a mean particle size of from about 0.05 to about 1.5 µm.

23. The formulation according to claim 22, wherein the micronized bioactive substance particles used exhibit a mean particle size of no more than a mean particle size of from about 0.1 to about 1Ø
µm.

24. The formulation according to claim 1 wherein the bioactive substance is an antimicrobial or antiviral antibiotic.

25. The formulation according to claim 24 wherein the antimicrobial antibiotic is selected from the group consisting of antibacterial antibiotics, antiparasitic antibiotics and antifungal antibiotics.

26. The formulation according to claim 25 wherein the antifungal antibiotic is selected from the group consisting of butocouazole nitrate, haloprogen, clotrimazole and other azoles.

27. The formulation according to claim 24 wherein the antiviral antibiotic is selected from the group consisting of O-[(2-hydroxyetoxy)-methyl]guanine anti-herpes medications, and tee tree oil.

28. The formulation according to claim 1 wherein the bioactive substance is a vitamin.

29. The formulation according to claim 28 wherein the vitamin is selected from the group consisting of vitamin C, vitamin E, vitamin B12, vitamin B6, folic acid, beta carotene and niacin and combinations thereof.

30. The formulation according to claim 29 wherein the vitamin is combination of vitamin C and vitamin E.

31. The formulation according to claim 28 wherein the vitamin is niacin and further comprises an anti-inflammatory agent.

32. The formulation according to claim 28 wherein the anti-inflammatory agent is selected from the group consisting of aspirin, ibuprofen, acetaminophen and cox-2 inhibitors.

33. The formulation according to claim 1 wherein the bioactive substance is a local anesthetic.

34. The formulation according to claim 1 wherein the bioactive substance is an antioxidant.

35. The formulation according to claim 34 wherein the antioxidant is an antioxidant derivative.

36. The formulation according to claim 35 wherein the antioxidant derivative is selected from the group consisting of esters, ethers, peptides, lipids, nucleotides and nucleosides, glycosyl, N-acetyl, methyl, ethyl, propyl, amyl, butyl lauryl, palmitoyl, oleyl, gamma-linoloyl,, cholesteryl and glyceryl esters of such antioxidants.

37. The formulation according to claim 34 wherein the antioxidant is selected from the group consisting of urocanic acid, imidazoles; D,L-carnosine, D-carnosine, L-carnosine, anscrine, peptides; alpha-carotine, beta-carotine, lycopine, carotines;
carotenoids; dihydrolipoic acid, lipoic acids;
aurothioglucose, propylthiouracil, thioredoxin, glutathion, cysteine, cystine, cystamine, thiols;
dilauryl thioproponate; distearyl thiopropionate;
thiopropionate; thiopropionic acid; butathione-sulfoxamines, homocysteine-sulphoxamine, butathione-sulphones, penta-, hexa- and heptathioninesulphomimine, sulfoxamine compounds;
alpha-hydroxy-fatty acids, palmitic acid, lactoferrin, EDTA, EGTA, metal chelating agents;
citric acid, lactic acid, malic acid, alpha-hydroxy acids; gamma-linolenic acid, linoleic acid, oleic acid, unsaturated fatty acids; folic acid; ubiquinone, ubiqinol and other quinones;
vitamin C; ascorbyl palmitate, Mg ascorbyl phosphate, ascorbyl acetate, vitamin C
derivatives; vitamin E acetate, tocopherols and tocopherol derivatives; vitamin A; vitamin A
palmitate, vitamin A derivatives; coniferyl benzooate of benzoin resin; rutic acid; alpha-glycosylrutin; ferulic acid furfurylideneglucitol;
carnosine; butylhydroxytoluene (BHT);
butylhydroxyanisole (BHA); nordihydroguaic resin acid; nordihydroguaiaretic acid;
trihydroxybuterophenone; uric acid; mannose; ZnO, ZnO4, zinc compounds; selenium and stilbenes.

38. The formulation according to claim 1 wherein the bioactive substance is a sugar, an amino acid or a small peptide.

39. The formulation according to claim 38 wherein the sugar, amino acid or small peptide is selected from the group consisting of carnosine; acetyl-L-carnithine, N-acetyl-carnithine, N-acetyl-cysteine, N-acetyl-D-glucosamine, B/D/L-alanine, alpha-keto-glutarates, arginates, L-arganine base/HCl, L-arginine-pyroglutamate, ascorbates, L-aspargine monohydrate, aspartame, aspartates, L-aspartic acid, L-carnithine base, L-carnithine fumarate, L-carnithine/HCl, L carnithine bitartrate, L-carnithine-L-tartrate, chelates, L-citruline, creatin monohydrate, creatine phosphate, creatine pyruvate, L-cysteine base, L-cysteine/HCl monohydrate, anhydrous L-cysteine/HCl, cysteinates, N,N-dimethylglycine base/HCl, glutamates, L-glutamic acid, L-glutamine, L-glutamine peptide, reduced L-glutathione, L-glycine, keto-glutaric acid, L-histidine base/HCl, L-isoleucine, L-leucine, lysinates, L-lysine monohydrate/HCl, D-mannose, DL/L-methionine, L-ornithine/HCl, DL/L-phenylalanine, L-hydroxyproline, trans-hydroxy-L-proline, pyroglutamic acid, D/L-ribose, L-selenium-methionine, L-serine, taurates, tartarates, L-threonine, trimethylglycine, DL/L-tryptophan, taurine, L-theanine, L-tyrosine, L-valine, xylitol, D-xylose,DL/L-zinc monomethionine, D or L-amino acids and short peptides and all bases, acids and salts thereof.

40. The formulation according to claim 1 wherein the bioactive substance is a biological additive.

41. The formulation according to claim 40 wherein the biological additive is a compound obtained from plants, animals, bacteria and yeast.

42. The formulation according to claim 41, wherein the biological additive is selected from the group consisting of tea tree oil, oil of Lavandula angustifolia, Carica papaya extract, Echinacea angustifolia extract, Mimosa tenuiflora extract, Hydrocotyl (centella) asiatica extract, gingko biloba extract, Matricaria chamomila (chamomile oil) extract, Hypericum perforatum extract, and Aloe barbedensis extract.

43. The formulation according to claim 42 wherein the biological additive is tea tree oil.

44. The formulation according to claim 40 wherein one or more biological additive is present in the formulation in a combined amount of from about 1%
to 10% by weight.

45. The formulation according to claim 44 wherein one or more biological additive is present in the formulation in a combined amount of from about 2%
to 8% by weight.

46. The formulation according to claim 45 wherein one or more biological additive is present in the formulation in a combined amount of from about 4%
to 6% by weight.

47. The formulation according to claim 1 wherein the bioactive substance is a sunblock.

48. The formulation according to claim 47 wherein the sunblock is not a UV absorber.

49. The formulation according to claim 47 wherein the sunblock is selected from the group consisting of dioxybenzone, ethyl 4-[bis(hydroxypropyl)]
aminobenzoate, glyceryl aminobenzoate, homosalate, menthyl anthranilate, octocrylene, octyl methoxycinnamate, octyl salicylate, oxybenzone, padimate O, red petrolatum, titanium dioxide, 4-methylbezylidene camphor, benzophenone-1, benzophenone-2, benzophenone-6, benzophenone-12, isopropyl dibenzoyl methane, butylmethoxydibenzoylmethane, zotocrylene, zinc oxide, para-aminobenzoic acid, cinnamate; and derivatives, analogs and functional analogs of such sunblocks and combinations thereof.

50. The formulation according to claim 1 wherein the exfoliants are chemical exfoliants.

51. The formulation according to claim 50, wherein the chemical exfoliant is AHA or BHA, or combinations thereof.

52. The formulation according to claim 1 wherein exfoliating agents are enzymes.

53. The formulation according to claim 52 wherein the exfoliating enzyme is selected from the group consisting of papain, is Linked-Papain.TM. (papain carbomer), serine proteases, macramidase, penicillinase, pepsin, plasmin, bromelains, streptokinase, sutilains, trypsin, urokinase, keratinase, amylase, hyaluronidase, cholic acid, chymopapain, chymotrypsin, cynara, brinolase and chenodeoxycholic acid.

54. The formulation according to claim 52 wherein the enzyme is present in the formulation in an amount between about 1% and 8%, by weight.

55. The formulation according to claim 54 wherein the enzyme is present in the formulation in an amount between about 2% and 5%, by weight.

56. The formulation according to claim 52 wherein the enzyme is present in the formulation in an amount of about 5%, by weight.

57. A method for treating a dermatologic condition, the method comprising applying the formulation according claim 1 at a therapeutically effective concentration to the skin of a patient having a dermatological condition.

58. The method according to claim 57, wherein the dermatologic condition is selected from the group consisting of acne, bruises, burns, eczema, mycoses, pruritis, psoriasis, seborrhea, scabs, shingles, tineapedis, wounds, wrinkles and erythema of acne rosacea.

59. The method according to claim 57 wherein the formulation includes an agent selected from the group consisting of anti-acne preparations; anti-inflammatory agents; monobenzone, depigmenting agents; amcinonide, diflorasone diacetate, hydrocortisone, dermatitis relief agents;
methylbenzethonium chloride, diaper rash relief agents; mineral oil, PEG-4 dilaurate, lanolin oil, petrolatum, mineral wax, emollients, moisturizers;
alclometasone dipropionate, betamethasone valerate, isopropyl myristate MSD, pruritic medications; anthralin, methoxsalen, coal tar, psoriasis, seborrhea and scabicide agents; 2-(acetyloxy)-9-fluoro-1',2'3'4'-tetrahydro-11-hydroxypregna-1,4-dieno[16,17b] napthalene-3,20-dione, 21-chloro-9-fluoro-1',2',3',4'-terahydro-11b-hydroxypregna-1,4-dieno[16z,17b] napthalene-3,20-dione and steroids.

60. A method for treating a dermatologic condition, the method comprising exfoliating the skin of a patient having a dermatological condition and applying to the skin a formulation comprising an anhydrous carrier medium and a high concentration of bioactive substance.

61. The method according to claim 60, wherein the dermatologic condition is selected from the group consisting of acne, bruises, burns, eczema, mycoses, pruritis, psoriasis, seborrhea, scabs, shingles, tineapedis, wounds, wrinkles and erythema of acne rosacea.

62. The method according to claim 60 wherein the formulation includes an agent selected from the group consisting of anti-acne preparations; anti-inflammatory agents; monobenzone, depigmenting agents; amcinonide, diflorasone diacetate, hydrocortisone, dermatitis relief agents;
methylbenzethonium chloride, diaper rash relief agents; mineral oil, PEG-4 dilaurate, lanolin oil, petrolatum, mineral wax, emollients, moisturizers;
alclometasone dipropionate, betamethasone valerate, isopropyl myristate MSD, pruritic medications; anthralin, methoxsalen, coal tar, psoriasis, seborrhea and scabicide agents; 2-(acetyloxy)-9-fluoro-1' ,2'3'4'-tetrahydro-11-hydroxypregna-1,4-dieno[16,17b] napthalene-3,20-dione, 21-chloro-9-fluoro-1',2',3',4'-terahydro-11b-hydroxypregna-1,4-dieno[16z,17b] napthalene-3,20-dione and steroids.

63. The method according to claim 60 wherein the formulation includes an exfoliant.

64. The method according to claim 63 wherein the exfoliation is mechanical.

65. The method according to claim 64 wherein the mechanical exfoliation is carried out by brushing, washing or particle pressure.

66. A method for introducing a bioactive substance into the bloodstream of a patient, the method comprising applying the formulation according to claim 1 to the skin of a patient.

67. A method for introducing a bioactive substance into the bloodstream of a patient, the method comprising exfoliating the skin of a patient and applying to the skin a formulation comprising an anhydrous carrier medium and a high concentration of bioactive substance.

68. The method according to claim 57 further including preventing the formulation according to claim 1 from escaping to the exterior of the skin.

69. The method according to claim 68, wherein the prevention is achieved by covering the area of the skin that has been treated with a formulation with an occlusive patch or other occlusive agent.

70. The method according to claim 60 further including preventing the formulation according from escaping to the exterior of the skin.

71. The method according to claim 70, wherein the prevention is achieved by covering the area of the skin that has been treated with a formulation with an occlusive patch or other occlusive agent.

72. The method according to claim 67 wherein the bioactive substance is an antimicrobial or antiviral antibiotic.

73. The method according to claim 72 wherein the antimicrobial antibiotic is selected from the group consisting of antibacterial antibiotics, antiparasitic antibiotics and antifungal antibiotics.

74. The method according to claim 73 wherein the antifungal antibiotic is selected from the group consisting of butocouazole nitrate, haloprogen, clotrimazole and other azoles.

75. The method according to claim 72 wherein the antiviral antibiotic is selected from the group consisting of O-[(2-hydroxyetoxy)-methyl]guanine anti-herpes medications, and tee tree oil.

76. The method according to claim 67 wherein the bioactive substance is a vitamin.

77. The method according to claim 76 wherein the vitamin is selected from the group consisting of vitamin C, vitamin E, vitamin B12, vitamin B6, folic acid, beta carotene and niacin and combinations thereof.

78. The method according to claim 77 wherein the vitamin is combination of vitamin C and vitamin E.

79. The method according to claim 78 wherein the vitamin is niacin and further comprises an anti-inflammatory agent.

80. The method according to claim 79 wherein the anti-inflammatory agent is selected from the group consisting of aspirin, ibuprofen, acetaminophen and cox-2 inhibitors.

81. The method according to claim 67 wherein the bioactive substance is an antioxidant.

82. The method according to claim 81 wherein the antioxidant is an antioxidant derivative.

83. The method according to claim 82 wherein the antioxidant derivative is selected from the group consisting of esters, ethers, peptides, lipids, nucleotides and nucleosides, glycosyl, N-acetyl, methyl, ethyl, propyl, amyl, butyl lauryl, palmitoyl, oleyl, gamma-linoloyl,, cholesteryl and glyceryl esters of such antioxidants.

84. The method according to claim 81 wherein the antioxidant is selected from the group consisting of urocanic acid, imidazoles; D,L-carnosine, D-carnosine, L-carnosine, anscrine, peptides; alpha-carotine, beta-carotine, lycopine, carotines;
carotenoids; dihydrolipoic acid, lipoic acids;
aurothioglucose, propylthiouracil, thioredoxin, glutathion, cysteine, cystine, cystamine, thiols;
dilauryl thioproponate; distearyl thiopropionate;
thiopropionate; thiopropionic acid; butathione-sulfoxamines, homocysteine-sulphoxamine, butathione-sulphones, penta-, hexa- and heptathioninesulphomimine, sulfoxamine compounds;
alpha-hydroxy-fatty acids, palmitic acid, lactoferrin, EDTA, EGTA, metal chelating agents;
citric acid, lactic acid, malic acid, alpha-hydroxy acids; gamma-linolenic acid, linoleic acid, oleic acid, unsaturated fatty acids; folic acid; ubiquinone, ubiqinol and other quinones;
vitamin C; ascorbyl palmitate, Mg ascorbyl phosphate, ascorbyl acetate, vitamin C
derivatives; vitamin E acetate, tocopherols and tocopherol derivatives; vitamin A; vitamin A
palmitate, vitamin A derivatives; coniferyl benzooate of benzoin resin; rutic acid; alpha-glycosylrutin; ferulic acid furfurylideneglucitol;
carnosine; butylhydroxytoluene (BHT);
butylhydroxyanisole (BHA); nordihydroguaic resin acid; nordihydroguaiaretic acid;
trihydroxybuterophenone; uric acid; mannose; ZnO, ZnO4, zinc compounds; selenium and stilbenes.

85. The method according to claim 67 wherein the bioactive substance is a sugar, an amino acid or a small peptide.

86. The method according to claim 85 wherein the sugar, an amino acid or small peptide is selected from the group consisting of carnosine, acetyl-L-carnithine, N-acetyl-carnithine, N-acetyl-cysteine, N-acetyl-D-glucosamine, B/D/L-alanine, alpha-keto-glutarates, arginates, L-arganine base/HCl, L-arginine-pyroglutamate, ascorbates, L-aspargine monohydrate, aspartame, aspartates, L-aspartic acid, L-carnithine base, L-carnithine fumarate, L-carnithine/HCl, L carnithine bitartrate, L-carnithine-L-tartrate, chelates, L-citruline, creatin monohydrate, creatine phosphate, creatine pyruvate, L-cysteine base, L-cysteine/HCl monohydrate, anhydrous L-cysteine/HCl, cysteinates, N,N-dimethylglycine base/HCl, glutamates, L-glutamic acid, L-glutamine, L-glutamine peptide, reduced L-glutathione, L-glycine, keto-glutaric acid, L-histidine base/HCl, L-isoleucine, L-leucine, lysinates, L-lysine monohydrate/HCl, D-mannose, DL/L-methionine, L-ornithine/HCl, DL/L-phenylalanine, L-hydroxyproline, trans-hydroxy-L-proline, pyroglutamic acid, D/L-ribose, L-selenium-methionine, L-serine, taurates, tartarates, L-threonine, trimethylglycine, DL/L-tryptophan, taurine, L-theanine, L-tyrosine, L-valine, xylitol, D-xylose,DL/L-zinc monomethionine, D or L-amino acids and short peptides and all bases, acids and salts thereof.

87. The method according to claim 67, wherein the formulation further comprises an exfoliant.

88. The method according to claim 67, wherein the exfoliation of the skin is chemical.

89. The method according to claim 67, wherein the exfoliation of the skin is mechanical.

90. The method according to claim 67, wherein the exfoliation of the skin is enzymatic.

91. The method according to claim 89, wherein the mechanical exfoliation of the skin is carried out by any mechanical frictional force, including brushing, washing or particle pressure.

92. The method according to claim 66, further including preventing the formulation according to claim 1 from escaping to the exterior of the skin.

93. The method according to claim 66, wherein the prevention is achieved by covering the area of the skin that has been treated with a formulation with an occlusive patch or other occlusive agent.

94. The method according to claim 67, further including preventing the formulation according to claim 1 from escaping to the exterior of the skin.

95. The method according to claim 67, wherein the prevention is achieved by covering the area of the skin that has been treated with a formulation with an occlusive patch or other occlusive agent.

96. The method according to claim 66 wherein the bioactive substance is an antimicrobial or antiviral antibiotic.

97. The method according to claim 96 wherein the antimicrobial antibiotic is selected from the group consisting of antibacterial antibiotics, antiparasitic antibiotics and antifungal antibiotics.

98. The method according to claim 97 wherein the antifungal antibiotic is selected from the group consisting of butocouazole nitrate, haloprogen, clotrimazole and other azoles.

99. The method according to claim 96 wherein the antiviral antibiotic is selected from the group consisting of 0-[(2-hydroxyetoxy)-methyl]guanine anti-herpes medications, and tee tree oil.

100. The method according to claim 66 wherein the bioactive substance is a vitamin.

101. The method according to claim 100 wherein the vitamin is selected from the group consisting of vitamin C, vitamin E, vitamin B12, vitamin B6, folic acid, beta carotene and niacin, or combinations thereof.

102. The method according to claim 100 wherein the vitamin is combination of vitamin C and vitamin E.

103. The method according to claim 100 wherein the vitamin is niacin and further comprises an anti-inflammatory agent.

104. The method according to claim 103 wherein the anti-inflammatory agent is selected from the group consisting of aspirin, ibuprofen, acetaminophen and cox-2 inhibitors.

105. The method according to claim 66 wherein the bioactive substance is an antioxidant.

106. The method according to claim 105 wherein the antioxidant is an antioxidant derivative.

107. The method according to claim 106 wherein the antioxidant derivative is selected from the group consisting of esters, ethers, peptides, lipids, nucleotides and nucleosides, glycosyl, N-acetyl, methyl, ethyl, propyl, amyl, butyl lauryl, palmitoyl, oleyl, gamma-linoloyl,, cholesteryl and glyceryl esters of such antioxidants.

108. The method according to claim 105 wherein the antioxidant is selected from the group consisting of urocanic acid, imidazoles; D,L-carnosine, D-carnosine, L-carnosine, anscrine, peptides; alpha-carotine, beta-carotine, lycopine, carotines;
carotenoids; dihydrolipoic acid, lipoic acids;
aurothioglucose, propylthiouracil, thioredoxin, glutathion, cysteine, cystine, cystamine, thiols;
dilauryl thioproponate; distearyl thiopropionate;
thiopropionate; thiopropionic acid; butathione-sulfoxamines, homocysteine-sulphoxamine, butathione-sulphones, penta-, hexa- and heptathioninesulphomimine, sulfoxamine compounds;

alpha-hydroxy-fatty acids, palmitic acid, lactoferrin, EDTA, EGTA, metal chelating agents;
citric acid, lactic acid, malic acid, alpha-hydroxy acids; gamma-linolenic acid, linoleic acid, oleic acid, unsaturated fatty acids; folic acid; ubiquinone, ubiqinol and other quinones;
vitamin C; ascorbyl palmitate, Mg ascorbyl phosphate, ascorbyl acetate, vitamin C
derivatives; vitamin E acetate, tocopherols and tocopherol derivatives; vitamin A; vitamin A
palmitate, vitamin A derivatives; coniferyl benzooate of benzoin resin; rutic acid; alpha-glycosylrutin; ferulic acid furfurylideneglucitol;
carnosine; butylhydroxytoluene (BHT);
butylhydroxyanisole (BHA); nordihydroguaic resin acid; nordihydroguaiaretic acid;
trihydroxybuterophenone; uric acid; mannose; ZnO, ZnO4, zinc compounds; selenium and stilbenes.

109. The method according to claim 66 wherein the bioactive substance is a sugar, an amino acid or a small peptide.

110. The method according to claim 109 wherein the sugar, an amino acid or small peptide is selected from the group consisting of carnosine, acetyl-L-carnithine, N-acetyl-carnithine, N-acetyl-cysteine, N-acetyl-D-glucosamine, B/D/L-alanine, alpha-keto-glutarates, arginates, L-arganine base/HCl, L-arginine-pyroglutamate, ascorbates, L-aspargine monohydrate, aspartame, aspartates, L-aspartic acid, L-carnithine base, L-carnithine fumarate, L-carnithine/HCl, L carnithine bitartrate, L-carnithine-L-tartrate, chelates, L-citruline, creatin monohydrate, creatine phosphate, creatine pyruvate, L-cysteine base, L-cysteine/HCl monohydrate, anhydrous L-cysteine/HCl, cysteinates, N,N-dimethylglycine base/HCl, glutamates, L-glutamic acid, L-glutamine, L-glutamine peptide, reduced L-glutathione, L-glycine, keto-glutaric acid, L-histidine base/HCl, L-isoleucine, L-leucine, lysinates, L-lysine monohydrate/HCl, D-mannose, DL/L-methionine, L-ornithine/HCl, DL/L-phenylalanine, L-hydroxyproline, trans-hydroxy-L-proline, pyroglutamic acid, D/L-ribose, L-selenium-methionine, L-serine, taurates, tartarates, L-threonine, trimethylglycine, DL/L-tryptophan, taurine, L-theanine, L-tyrosine, L-valine, xylitol, D-xylose,DL/L-zinc monomethionine, D or L-amino acids and short peptides and all bases, acids and salts thereof and combinations thereof.

111. The method according to claim 66, wherein the exfoliation of the skin is chemical.

112. The method according to claim 66, wherein the exfoliation of the skin is mechanical.

113. The method according to claim 66, wherein the exfoliation of the skin is enzymatic.

114. The method according to claim 66, wherein the mechanical exfoliation of the skin is carried out by any mechanical frictional force, including brushing, washing or particle pressure.

115. The method according to claim 78, wherein the formulation further comprises niacin.

116. The method according to claim 102, wherein the formulation further comprises niacin.

117. The formulation according to claim 30, further comprising niacin.