Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
  • A61Q19/00—Preparations for care of the skin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
  • A61P17/16—Emollients or protectives, e.g. against radiation
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
  • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
  • A61K2800/57—Compounds covalently linked to a(n inert) carrier molecule, e.g. conjugates, pro-fragrances
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
  • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
  • A61K2800/94—Involves covalent bonding to the substrate

Definitions

• This invention relates to the linkage of agents to tissue by certain reactive moieties and involves methods, products and kits relating thereto.
• Transglutaminases are a family of calcium-dependent enzymes mediating covalent cross-linking reactions between specific peptide bound (-glutamyl residues and various primary amino groups of peptide-bound ly sines or polyamines, acting as amine donor substrates (Davies, et al., Adv. Exp. Med. Biol. 250, 391-401, 1988). These enzymes stabilize biological structures via the formation of isopeptide cross-links. In mammals, at least five enzymatically active transglutaminases have been identified, cloned and sequenced. The number of proteins acting as glutaminyl substrates for transglutaminases is restricted, and no obvious consensus sequence around these substrates' glutamines has been found.
• transglutaminases and corneocyte proteins the natural substrates of transglutaminases, can be used together as cosmetic treatments to cross-link preparations of corneocyte proteins to the outer layer of skin, hair or nails to form a protective layer on the skin, hair or nails.
• Patent 5,490,980 describes selecting agents having or modifying agents to have an aliphatic amine, and then attaching those agents to skin, hair or nails using transglutaminase. While the idea was sound in principle, in practice the '980 applicants achieved results that were barely above background. (See Example Section of '980 patent). An aliphatic amine was applied in the examples as a single linking molecule or prophetically in clusters (according to a formula in the '980 patent).
• composition of matter comprising a compound having a structure of Formula I
• A is an agent
• Li and L 2 are independently selected organic linkers (i.e., linking molecules) or bonds
• Xi and X are reactive moieties independently selected from Group A reactive moieties, and, in some instances, N-hydroxyl succinimide or N-alkyl maleimide, and wherein L 2 and X 2 may be present or absent, however if L 2 is absent, then X 2 is also absent.
• Compounds of Formula I can be both monofunctional, provided X] or X 2 , but not both are present, or if both are present, the compound is bifunctional.
• the agent is provided in a microparticle.
• the reactive moiety is not native to the agent.
• the reactive moieties are dihydroxyacetone.
• Preferred monofunctional reactive moieties which can be linked to the agent are as described herein.
• the monofunctional reactive moieties of Group A are most particularly suited for linkage to non-protein or non-peptide based agents. Since many of these reactive moieties react with amines (such as those present in lysines) or thiols (such as those present in cysteines), it is recommended that the agent to which they attach possesses neither amines nor thiols. This will prevent the reactive moieties from reacting with the agent rather than the intended body tissue.
• Some preferred monofunctional reactive moieties are shown below in Group A. The
• R group may be any organic or inorganic molecule.
• R may include the agent or a microparticle comprising the agent.
• Group A Reactive Moieties
• the agent, A is selected from the group consisting of a sunscreen agent, a cosmetic, an enzyme, a coloring agent, a pharmaceutical agent, a member of a ligand/receptor pair, a tissue sealant, a wound healing agent, a bulking agent, a hair conditioning agent, a hair fixative, an anti-foaming agent, a moisturizing agent, a humectant, a depilatory agent, an anti-nerve gas agent including an enzyme that degrades nerve agents, an anti-neurotoxin, a film forming agent, a vitamin, an insect repellant and a component of a high affinity noncovalent coupling (e.g., biotin or avidin).
• a sunscreen agent e.g., a cosmetic, an enzyme, a coloring agent, a pharmaceutical agent, a member of a ligand/receptor pair, a tissue sealant, a wound healing agent, a bulking agent, a hair conditioning agent, a hair fixative, an anti
• A is not a protein or a peptide or does not contain amine or thiol groups (as are present in lysine and cysteine, respectively). In some embodiments, A is not an anti-nerve gas agent.
• the agent is selected from the group consisting of cholinesterase and phosphodiesterase.
• the agent is an enzyme that degrades nerve agents, such as organophosphate agents.
• the agent is selected from the group consisting of OPAA anhydrolase (prolidase) and squid type OPA anhydrase.
• the agent is a nonprotein.
• the agent is not itself able to conjugate directly to proteinaceous material or conjugates only weakly.
• the agent is conjugated to a reactive moiety whereby the agent may be attached to the body tissue via the covalent bond resulting between the reactive moiety and the proteinaceous material.
• a proteinaceous material is minimally defined as a material that contains thiols and/or amines and to which the reactive moieties of the invention may covalently attach in a spontaneous manner.
• the preferred proteinaceous material is body tissue, including the integument, a wound bed, internal organs or internal tissue of a living subject.
• the agent in its native form e.g., free of conjugation to either the linker (i.e., linking molecule) or the reactive moiety
• the linker when not conjugated to the reactive moiety is not able to covalently attach to a body tissue spontaneously.
• the reactive moiety is dihydroxy acetone.
• Xi, X 2 or both Xj and X 2 may be dihydroxyacetone, according to some embodiments.
• the invention provides a method for attaching an agent to a body tissue comprising applying to the body tissue a compound having a structure of Formula I in an effective amount, and allowing crosslinking to occur.
• the conjugate of agent, linker and reactive moiety i.e., the compound of Formula I
• crosslinking may occur with no additional physical requirements or with additional physical requirements such as light, heat, catalysts, etc.
• a body tissue can be an integument, skin, hair and nails, a wound bed, or an internal body tissue.
• the compound of Formula I to be used in this method may incorporate a cosmetic agent, a bulking agent, a hair conditioning agent, a hair fixative, a sunscreen agent, a moisturizing agent, a depilatory agent, an anti-nerve gas agent, a film forming agent, a vitamin, an insect repellant, a coloring agent, a pharmaceutical agent, a ligand-receptor complex or a receptor of a ligand-receptor complex, although agents which do not possess amine or thiol groups are generally preferred when using the monofunctional reactive moieties of Group A.
• the invention may be used for a number of different purposes depending upon the nature of the particular agent used in the compositions and methods.
• the agents, the linkers or the agent-linker conjugates may be provided in the form of a microparticle. In yet other embodiments, the agents, the linkers or the agent-linker conjugates do not comprise a microparticle.
• the invention therefore also provides a method for coloring hair by applying to hair a compound of Formula I, wherein the agent is a coloring agent such as those described herein, in an effective amount to change the color of untreated hair.
• the invention provides a method for moisturizing skin by applying to the skin a compound having a structure of Formula I, wherein the agent is a moisturizing agent as described herein, in an effective amount to improve hydration of skin.
• the invention provides methods for bulking hair, for conditioning hair, for attaching a sunscreen agent, for topically applying a pharmaceutical, etc.
• the tissue can be pretreated to make it more receptive to crosslinking with the reactive moiety. In one embodiment described above, this is accomplished by attaching polymers rich in reactive molecules such as glutamine, lysine or both glutamine and lysine to the body tissue. In other embodiments, the tissue is treated to expose reactive molecules by washing, chemical treatment, etc. Detergents and Upases can be used to remove fatty acids and oils. Roughening agents such as pumice, silica and sandpaper can be employed to remove dead tissue and other obstructions, and chemical agents such as sodium hydroxide can be used to expose reactive molecules. Combinations of the foregoing arc contemplated.
• the invention provides monofunctional reactive compounds (e.g., where either X
• the invention further provides compositions and methods of use of other bifunctional reactive compounds which do not comprise an agent.
• These bifunctional reactive compounds have the structure of Formula II:
• L is an organic linker or bond
• and X 2 arc reactive moieties independently selected from the reactive moieties of Group A, N-hydroxy-succinimide and N-alkylmaleimide and derivatives thereof, as well as any other reactive moiety described herein.
• Compounds of Formula II embrace the structures of bis-N-hydroxy-succinimide or bis-N-alkyl maleimide shown below.
• Bifunctional reactive molecules can be used to link a tissue to an agent, or to link a tissue to a microparticle, or to link a microparticle to an agent, or to link to two tissues together, but is not so limited. Some bifunctional reactive molecules can also deliver an agent. This latter group of molecules include compounds of Formula I where both X] and X 2 are present and compounds of Formula III where both Xi, X 2 , and any of A], A 2 or A 3 are present.
• the invention provides a method for sealing tissue comprising applying a force to hold two tissues in contact with each other in the presence of an effective amount of a compound of Formula II to crosslink the two tissues, and allowing crosslinking to occur.
• these latter compounds are used to glue tissues together.
• the tissue may be held together during the crosslinking process by any conventional means, such as sutures, tape, stapes and the like.
• Formula I compounds in which L ls Xi, L 2 , and X 2 are all present can also be used to link two tissues together in a similar manner as Formula II compounds. This latter category of Formula I compounds are also able to provide the two tissues with an agent while sealing them together.
• derivatives of Formula II compounds which are linked to an agent can also seal two tissues and provide them with an agent.
• compositions, methods and kits that include compounds that are derivatives of Formula II and which have the structure of Formula III:
• Al, A2, and A3 are agents
• L is an organic linker and Xi and X 2 are reactive moieties independently selected from Group A, N-hydroxy-succinimide, N-alkyl maleimide, and derivatives thereof; wherein either Ai or A 2 may be present, but not both, and wherein A 3 is present independently of the presence of Aj or A 2 .
• Any or all of Aj, A 2 and A 3 may be a microparticle.
• Ai is absent; A 2 is absent; A 3 is absent; Aj and A 3 are absent; A] and A are absent; A 2 and A 3 are absent; or Aj, A 2 and A 3 are absent, thereby reverting the Formula III compound to a Formula II compound.
• the invention provides a method for attaching an agent to a body tissue comprising applying to the body tissue a compound having a structure of Formula II in an effective amount, applying to the body tissue an agent, and allowing crosslinking to occur.
• the agent may be applied to the body tissue prior to, simultaneously with, or following the application of the Formula II compound.
• a body tissue can be an integument, skin, hair and nails, a wound bed, or an internal body tissue.
• the compounds of Formula II (including the subset described above) to be used in this method may incorporate a cosmetic agent, a bulking agent, a hair conditioning agent, a hair fixative, a sunscreen agent, a moisturizing agent, a depilatory agent, an anti-nerve gas agent, a film forming agent, a vitamin, an insect repellant, a coloring agent, a pharmaceutical agent, a ligand-receptor complex or a receptor of a ligand-receptor complex.
• the bifunctional reactive molecules e.g., compounds of Formula II
• These bifunctional compounds can be used with agents which possess amines (such as are present in lysine) or thiols (such as are present in cysteines).
• the invention therefore also provides a method for coloring hair by applying to hair a compound of Formula II, wherein the agent is a coloring agent such as those described herein, in an effective amount to change the color of untreated hair.
• the invention provides a method for moisturizing skin by applying to the skin a compound having a structure of Formula II, wherein the agent is a moisturizing agent as described herein, in an effective amount to improve hydration of skin.
• the invention provides methods for bulking hair, for conditioning hair, for attaching a sunscreen agent, for topically applying a pharmaceutical, etc.
• the invention in another aspect, provides a pharmaceutical composition comprising an effective amount of a compound of Formula I where A is a pharmaceutical agent, and a pharmaceutically acceptable carrier.
• the invention provides a pharmaceutical composition comprising an effective amount of a compound of Formula II (which may optionally comprise an agent) and a pharmaceutically acceptable carrier.
• the invention in another aspect, provides cosmetic compositions, skin protective compositions, nerve-gas deactivating compositions, etc.
• the agent which may comprise a compound in which the agent is a cosmetic agent, a bulking agent, a hair conditioning agent, a hair fixative, a sunscreen agent, a moisturizing agent, a depilatory agent, an anti-nerve gas agent, a film forming agent, a vitamin, an insect repellant, a coloring agent, a ligand-receptor complex or a receptor of a ligand-receptor complex.
• a kit comprising a package which houses a container containing a compound of Formula I, Formula II or Formula III and instructions for use.
• the container also can contain catalysts, microparticles, cleansers, vehicles, preservatives and buffers.
• the compounds of Formula I comprise dihydroxyacetone as the reactive moiety.
• a method is provided of treating a subject to attach microparticles to a skin surface of the subject comprising contacting the skin surface with microparticles having surface available reactive moieties in an amount sufficient to attach the microparticles to the skin surface, and allowing the microparticles to remain in contact with the skin surface for a time sufficient to permit a layer of microparticles to covalently attach to the skin surface.
• the surface available reactive moieties are selected from the group consisting of reactive moieties of Group A, N-hydroxy-succinimide and N-alkyl-maleimide, derivatives thereof, other reactive moieties mentioned herein.
• the layer of microparticles is non-planar.
• the microparticles further comprise an agent, or an active agent, or a non-nucleic acid active agent, or a non-protein active agent.
• the agent may be, but need not be, selected from the group consisting of a cosmetic agent, a bulking agent, a hair conditioning agent, a hair fixative, a sunscreen agent, a moisturizing agent, a depilatory agent, an anti-nerve gas agent, a film forming agent, a vitamin, an insect repellant, a coloring agent, a pharmaceutical agent, a ligand-receptor complex and a receptor of a ligand-receptor complex.
• the agent does not itself contain a reactive moiety.
• the microparticles further comprise a synthetic polymer, and in preferred embodiments, the synthetic polymer is latex or polystyrene. In other embodiments, the microparticles are non-biodegradable, water insoluble and/or detergent insoluble.
• the microparticles may be hollow or porous but are not so limited.
• the microparticles' size is selected from the group consisting of greater than 5 ⁇ m, less than 5 ⁇ m, less than 1 ⁇ m, 100 nm to 500 nm, less than 100 nm, 20 nm to 90 nm, 20 nm to 35 nm, less than 20 nm, 1 nm to 10 nm and 5 nm to 10 nm. These sizes and ranges can be cut-offs or can represent average size determinations.
• the microparticles enter the cornified layer of the skin but not the layer of living cells.
• the agent contained within the microparticle may be able to enter the layer of living cells.
• the reactive moieties are part of a polymer, preferably wherein the polymer is covalently attached to the microparticle.
• the polymer is comprised of at least 20%, at least 30%, at least 40%, at least 50% units having reactive moieties, or is rich in units having reactive moieties at a surface available terminus, or is selected from the group of polymers consisting of: at least two contiguous linked units having reactive moieties, at least three contiguous linked units having reactive moieties, at least four contiguous linked units having reactive moieties, at least five contiguous linked units having reactive moieties, at least ten contiguous linked units having reactive moieties, at least fifteen contiguous linked units having reactive moieties, and at least twenty contiguous linked units having reactive moieties.
• compositions, methods, and kits provided by the invention including those relating to microparticles.
• the invention provides a method of treating a subject to attach microparticles (or an agent) to a skin surface of the subject comprising contacting the skin surface with a bifunctional reactive compound of Formula I, II or III, contacting the skin with microparticles (or an agent) having surface available amines or thiols in an amount sufficient to attach the microparticles (or an agent) to the skin surface in the presence of the bifunctional reactive compound of Formula I, II or III, and allowing the microparticles (or an agent) and compound of Formula I, II or III to remain in contact with the skin surface for a time sufficient to permit a layer of microparticles (or an agent) to covalently attach to the skin surface.
• the surface available reactive amines or thiols are present as lysines or cysteines, respectively.
• the reactive amines or thiols are part of a polymer, and the polymer may be covalently attached to the microparticle (or an _ J J _
• the polymer is comprised of at least 50% amines, at least 50% thiols, at least 50% lysines, or at least 50% cysteines, or the polymer is amine-rich or lysine-rich at a surface available terminus, or it is thiol-rich or cysteine-rich at a surface available terminus, or it comprises a polymer selected from the group consisting of: at least two contiguous linked amines, lysines, thiols, or cysteines, at least three contiguous linked amines, lysines, thiols, or cysteines, at least four contiguous linked amines, lysines, thiols, or cysteines, at least five contiguous linked amines, lysines, thiols, or cysteines, at least ten contiguous linked amines, lysines, thiols, or cysteines, at least ten contiguous linked amines, lysines,
• the invention provides a composition comprising a microparticle comprising an active agent and a polymer rich in reactive molecules, wherein the microparticle is non-biodegradable, and the reactive molecules are surface available.
• the active agent is a nucleic acid agent, or a non-nucleic acid active agent, or a protein agent, or a non-protein active agent, or the active agent is selected from the group consisting of a cosmetic agent, a bulking agent, a hair conditioning agent, a hair fixative, a sunscreen agent, a moisturizing agent, a depilatory agent, an anti-nerve gas agent, a film forming agent, a vitamin, an insect repellant, a coloring agent, a pharmaceutical agent, a ligand-receptor complex and a receptor of a ligand-receptor complex.
• the active agent does not itself contain a reactive molecule.
• the microparticle further comprises a synthetic polymer and preferably the synthetic polymer is latex or polystyrene. In some embodiments, the polymer rich in reactive molecules is covalently linked to the synthetic polymer.
• the microparticles' size is selected from the group consisting of greater than 5 ⁇ m, less than 5 ⁇ m, less than 1 ⁇ m, 100 nm to 500 nm, less than 100 nm, 20 nm to 90 nm, 20 nm to 35 nm, less than 20 nm, 1 nm to 10 nm and 5 nm to 10 nm.
• the microparticle is non-biodegradable, water insoluble and/or or detergent insoluble.
• the reactive molecules are surface available in an amount sufficient to attach the microparticle to a skin surface in the presence of a compound of Formula I, II and III.
• the polymer comprises units and at least 50% of units have reactive moieties, or the polymer is rich in reactive moieties at a surface available terminus, or the polymer is covalently attached to the microparticle, or the polymer comprises units having reactive moieties wherein the polymer is selected from the group consisting of at least two contiguous linked units having reactive moieties, at least three contiguous linked units having reactive moieties, at least four contiguous linked units having reactive moieties, and at least five contiguous linked units having reactive moieties.
• the invention provides a composition comprising a microparticle comprising a non-nucleic acid active agent, and covalently attached surface available reactive molecules, wherein the microparticle is 100 nm to 500 nm in size. Other sizes are as described above.
• the surface available reactive moieties are present as free pendant groups.
• the invention provides a kit comprising any of the microparticles of the invention.
• the kit includes a microparticle comprising surface available reactive moieties in an amount sufficient to attach the microparticle to a skin surface, and instructions for topically administering the microparticle to a skin surface, wherein the surface available reactive moieties are selected from the group consisting of reactive moieties of Group A, N-hydroxyl-succinimide, and N-alkyl-maleimide, and derivatives thereof, and the like.
• the kit may further comprise a polylysine polymer for attachment to the skin surface, in one embodiment. In another embodiment, the kit may further comprise a cleanser.
• the microparticle is provided in a topically administered form selected from the group consisting of an ointment, an aerosol, a gel, and a lotion.
• the kit further comprises an agent in a separate container.
• kits comprising a bifunctional reactive molecule in an amount sufficient to crosslink a skin surface, and a microparticle comprising surface available reactive moieties in an amount sufficient to attach the microparticle to the bifunctional reactive molecule, wherein the surface available reactive moieties are selected from the group consisting of amines and thiols.
• the kit further comprises instructions for topically administering the bifunctional reactive molecule and the microparticle to a skin surface.
• the kit further comprises a cleanser.
• reactive moieties that are photoreactive may also be used in the compounds of Formula I, II and III.
• attachment of such reactive moieties to body tissues require the application of light of a particular wavelength, as will be described herein.
• Figure 1 is a schematic of a kit of the invention.
• the invention is based in part on the discovery that compounds bearing reactive carbonyl and hydroxyl groups are particularly useful for attaching agents to proteinaceous material such as body tissues, including external surfaces such as skin, hair, and nails.
• proteinaceous material such as body tissues, including external surfaces such as skin, hair, and nails.
• One category of such agent-bearing compounds have the structure of Formula I:
• A is an agent
• Lj and L 2 are independently organic linkers or bonds
• Xi and X 2 are reactive moieties independently selected from Group A, N-hydroxyl succinimide or N-alkyl maleimide, derivatives thereof, as well as any of the other reactive molecules described herein
• L and X 2 may be present or absent, however if L 2 is absent, then X 2 is also absent.
• the agent may be provided in the form of a microparticle.
• reactive moiety refers to the reactive chemical group or structure which reacts with its counterpart to form crosslinks.
• reactive moieties useful in the invention include the reactive carbonyl groups and Michael acceptors of Group A.
• a reactive molecule comprises one or more reactive moieties.
• Compounds of Formula I, Formula II, and Formula II are all reactive molecules.
• Group A reactive moieties contain reactive moieties which covalently attach to proteinaceous materials.
• Proteinaceous materials refer to materials which possess at least thiol and/or amine groups, and preferably comprise amino acids or peptides containing thiol and/or amine groups.
• Some Group A compounds, such as compounds (A), (B), (C) and (F) possess a reactive carbonyl group, while others, such as compounds (D) and (M) possess reactive Michael acceptors.
• reactive carbonyl groups attack amine groups such as those present in lysine residues.
• 1, 2 -dihydroxy acetone undergoes oxidative transformations in situ to generate cross-links with endogenous amines, including the ⁇ -amino group of lysine.
• Reactive Michael acceptors are thiol reactive, and thus preferentially target cysteine residues.
• Other suitable reactive compounds useful in the invention include imidoesters, active halogen compounds, EDC coupled compounds, pyridyl disulfide compounds, vinyl-sulfone containing compounds, and isocyanate containing compounds.
• Photoreactive moieties are able to modify epidermal proteins as a consequence of light activation. Although chemically unreactive in the absence of light, these reagents become transiently reactive upon illumination by wavelengths of light within their absorption spectrum. Suitable wavelengths are typically in the 300-500 nm range. These reagents are generally highly photosensitive and, consequently, are readily photoactivated.
• the reactive moiety is a photoreactive moiety which is linked to an agent either directly or indirectly through the use of a linker.
• the resultant reactive molecule can be applied to the epidermis (i.e., skin), nails or hair, followed by limited exposure to an appropriate wavelength. Light damage to the skin is probably unlikely given the enormous photosensitivity of these molecules.
• Photoreactive modifying reactive moieties are well known to those of ordinary skill in the art. Examples of such compounds include phenyl azides, benzophenones, diazo esters/amides and trifluoromethyl diazo esters/amides. Due to the high reactivity of these compounds, they are able to covalently attach to a variety of groups, including but not limited to amines and thiols. Upon appropriate light exposure, phenyl azides release a N 2 and present a free nitrogen radical (see Group A, compound E). Benzophenones on the other hand, convert their carbonyl group to a free radical (see Group A, compound F). Diazo esters/amides (Group A, compound G) and trifluomethyl diazo esters/amides (Group A, compound H) both form diradical carbons by releasing N 2 following exposure to light.
• the site of action for the compounds of the invention will depend on the extent to which these compounds can penetrate the surfaces or tissues to which they are applied.
• the action of an agent conjugated to the skin via dihydroxyacetone is generally limited to the upper layers of the skin.
• the reaction time and conditions necessary to effect a covalent attachment will differ. Dihydroxyacetone interaction with skin generally requires about 2 hours for optimal covalent attachment. However, conditions may be varied so as to reduce this reaction time. Such variables are well known to one of ordinary skill in the art.
• any of the reactive moieties discussed herein, including derivatives thereof may be incorporated into the compounds of Formula I, II and III, as appropriate.
• reactive groups which are useful in Formula I or II or III, these moieties may be suitable in all three of these formulae.
• the agents are chemical agents and include, but are not limited to, pharmaceutical agents, enzymes, cosmetics, bulking agents, hair conditioner agents, hair fixative agents, anti-foaming agents, antistatic agents, moisturizing agents including humectants, depilatories (i.e., hair removal agents), vitamins, film forming agents such as those used in hair fixatives or wound healing, anti-nerve gas or anti-neurotoxin agents, sunscreen agents, ligands of ligand-receptor pairs, receptors of ligand-receptor pairs, components of high affinity noncovalent bonding pairs, insecticides and repellants including louse repellents, bactericides, fungicides, tissue sealants, labels, structural proteins, chelating agents, microparticles and the like.
• Anti-nerve gas agents include enzymes which degrade nerve agents (e.g., organophosphate nerve agents). Examples include OPAA anhydrolase (prolidase) and the squid type OP A anhydrase. Other anti-nerve agents include pyridostigmine and pralidoxime chloride. Examples of agents useful in the invention are listed below.
• active agent it is meant that the agent, once coupled to a biological tissue in vivo or in vitro, has, maintains or can be released to have a desired activity such as a desired physiological activity or therapeutic activity.
• active agents are pharmaceutical agents, sunscreen agents, insecticides, bactericides, fungicides, etc.
• the active agent is not a cosmetic agent.
• the active agent is not a labeling agent such as a diagnostic agent.
• Compounds of Formula I are intended for a variety of uses, including but not limited to agent delivery and tissue sealing, provided their application to a body tissue is non-toxic. According to the invention, the agents are linked to proteinaceous material.
• the agents are preferably conjugated to a body tissue via the reactive molecule(s).
• a body tissue particularly important body tissues as sites of attachment are the integument (including specifically skin, nails, hair, mucous membranes and the surface of the eye), internal organs, internal tissue and wound beds.
• the tissue may be a body tissue, a tissue or cell isolate, isolated proteins, synthetic proteins, cell cultures and the like for use, for example, in assay systems according to the invention.
• conjugates of agents and reactive moieties are applied, for example, to a body tissue.
• the agents are then covalently linked to that tissue through the reactive moieties.
• Reactive moieties useful in the invention are those of Group A (and derivatives thereof), N-hydroxyl succinimide and N-alkyl maleimide, all of which can be used as monofunctional reactive molecules.
• reactive molecules useful to the invention are those which are bifunctional, examples of which include, but are not limited to, bis-N-hydroxy-succinimide and bis-N- alkyl-maleimide.
• Formula II compounds, which include these latter two compounds generally have the following structure:
• Xi and X 2 are reactive moieties independently selected from Group A, N-hydroxy- succinimide, and N-alkyl-maleimide (or derivatives thereof); and L is a linking molecule. Reactions of N-hydroxy-succinimide and N-alkyl-maleimide with their respective targets are shown below:
• a "bifunctional reactive molecule” or a "bifunctional reactive compound” is one which possesses at least two reactive moieties.
• the reactive moieties need not be the same and they may react with different groups.
• Bifunctional reactive compounds may take the form of Formula I, Formula II, and Formula III compounds, provided both Xi and X 2 are present.
• Formula I and Formula III compounds embrace bifunctional reactive compounds which are capable of agent delivery.
• Formula II compounds are generally not used for agent delivery.
• All bifunctional reactive molecules are capable of linking two separate entities, such as for example two tissues, a microparticle and a tissue, or an agent and a tissue.
• the agents used according to the invention are not themselves, in their native form, able to covalently attach to proteinaceous materials.
• a conjugate means two entities stably bound to one another by any physiochemical means. It is important that the nature of the attachment be of such a nature that it does not impair substantially the effectiveness of the agent or the substrate binding ability of the linking molecule, or the reactivity of any remaining reactive molecule. Keeping these parameters in mind, any linkage known to those of ordinary skill in the art may be employed including covalent or noncovalent. Covalent linkage is preferred. Such means and methods of attachment are well known to those of ordinary skill in the art.
• linker which can be used to tether or simply join the agent (which may be provided as a microparticle) and the reactive moiety.
• linker and “linking molecule” are used interchangeably.
• One embodiment involves linkers that are polymers.
• the polymer can be a homopolymer or a heteropolymer.
• spacers can be any of a variety of molecules, preferably nonactive, such as straight or even branched carbon chains of C]-C 3 o, saturated or unsaturated, phospholipids, amino acids, and in particular glycine, and the like, naturally occurring or synthetic. Additional spacers include alkyl and alkenyl carbonates, carbamates, and carbamides. These are all related and may add polar functionality to the spacers such as the C ⁇ -C 30 previously mentioned.
• linkers of Formulae I, II and III may be spacers.
• the conjugations or modifications described herein employ routine chemistry, which chemistry does not form a part of the invention and which chemistry is well known to those skilled in the art of chemistry.
• the use of protecting groups and known linkers such as mono and heterobifunctional linkers are well documented in the literature and will not be repeated here.
• Attachment of the reactive moiety to the agent (or the microparticle) according to the invention thus need not be direct attachment.
• the components X and A of the compositions of the invention may be provided with functionalized groups to facilitate their attachment to one another and/or linker groups may be interposed between these components to facilitate their attachment.
• the components of the compositions of the present invention may be synthesized in a single process, whereby the components could be regarded as one and the same entity.
• the linker molecule thus may contain functional groups for joining the X to the A, selected from the group consisting of a carboxylate group, an amino group, a sulfhydryl group, an imidazole group, an alkene group (a carbon atom double bonded to another carbon atom), an acyl halogen group, e.g., an acylchloride, and CH 2 X, wherein X represents a halogen (e.g., the two binding moieties are linked when a nucleophilic group displaces the halogen from the functional group of the CH 2 X linker molecule).
• the linker molecules may be of varying length spacer segments (linker molecules).
• Bifunctional linker molecules are also useful in the invention.
• a wide assortment of dicarboxylic spacer linker molecules are commercially available (see below for discussion). This includes linker molecules which have various internal heteroatoms and other functional groups, in addition to the terminal carboxylic groups, e.g., ethylene glycobissuccinate.
• linker ethylene glycobissuccinate (instead of adipic acid) provides a bivalent compound with a spacer of about twice the length of the adipoyl moiety. Also, the internal heteroatoms may confer improved water solubility over a straight chain hydrocarbon of similar length.
• Imidoester homobifunctional cross-linkers were among the first used to immobilize proteins onto solid-phase supports. They were used extensively for the study of protein structure and molecular associations in membranes. Although these cross-linkers are still used in protein subunit studies and solid-phase immobilization, they have been steadily replaced by the more stable, more efficient homobifunctional NHS-ester cross-linkers. Homobifunctional imidoesters maintain the net electronic charge on protein after cross- linking. Spacer arm lengths range from about 8.6 A to about 11.9 A. Imidoester cross-linkers react rapidly with amines at alkaline pH, but they have short half lives.
• Imidoesters are also very useful for protein-protein cross-links. These cross-linkers can penetrate cell membranes and cross-link proteins within the membrane to study membrane composition, structure and protein-protein and protein-lipid interactions. Imidoesters are also useful for oligomer formation. For example, cross-linking proteins to form oligomers may reveal if a bivalent, dimeric or trimeric form of the protein is responsible for activity. 2. N-Hydroxysuccinimide-Esters (NHS-esters)
• NHS-esters yield stable products upon reaction with primary or secondary amines. Coupling is efficient at physiological pH, NHS-ester cross-linkers are also more stable in solution than their imidate counterparts. Homobifunctional NHS-ester conjugations are commonly used to cross-link amine-containing proteins in either one-step or two-step reactions.
• Coupling through sulfhydryl groups is advantageous because it can be site-directed, yield cleavable products and allow for sequential coupling.
• a protein in a complex mixture can be specifically labeled if it is the only one with a free sulfhydryl group on its surface, a.
• the maleimide group is specific for sulfhydryl groups when the pH of the reaction mixture is kept between pH 6.5 and 7.5. At pH 7, the reaction of the maleimides with sulfhydryls is 1000-fold faster than with amines. Maleimides do not react with tyrosines, histidines or methionines. b. Haloacetyls
• the most common used ⁇ -Haloacetyls react with sulfhydryl groups at physiological pH.
• the reaction of the iodoacetyl group with a sulfhydryl proceeds by nucleophilic substitution of iodine, with a thiol producing a stable thioether linkage.
• Selectivity for sulfhydryl groups is ensured by using only a slight excess of the iodoacetyl group over the number of sulfhydryl groups at pH 8.3.
• iodoacetyl group can react with other amino acids.
• Pyridyl Disulfides Pyridyl disulfides react with sulfhydryls groups to form a disulfide bond. Pyridine-2- thione is released as a by-product of this reaction. These reagents can be used as cross-linkers and to introduce sulfhydryl groups into proteins.
• Carbodiimides couple carboxyls to primary amines or hydrazides, resulting in formation of amide or hydrazone bonds.
• Carbodiimides are unlike other conjugation reactions in that no cross-bridge is formed between the molecules being coupled.
• Carboxy termini of proteins can be targeted, as well as glutamic and aspartic acid side chains.
• polymerization is likely to occur because proteins contain carboxyls and amines.
• No cross-bridge is formed, and the amide bond is the same as a peptide bond, so reversal of the cross-linking is impossible without destruction of the protein.
• EDC Pulierce Co.
• a photoaffinity reagent is a compound that is chemically inert but becomes reactive when exposed to ultraviolet or visible light.
• Arylazides are photoaffinity reagents that are photolyzed at wavelengths between 250-460 nm, forming a reactive aryl nitrene. The aryl nitrene reacts nonselectively to form a covalent bond. Reducing agents must be used with caution because they can reduce the azido group.
• Glyoxals are useful compounds for targeting the guanidinyl portion of arginine residues. Glyoxals will target arginines at mildly alkaline pH. There is some cross-reactivity (the greatest at higher pH) with lysines. b. Carbonyl Specific Cross-Linkers
• Carbonyls (aldehydes and ketones) react with amines and hydrazides at pH 5-7.
• the reaction with hydrazides is faster than with amines, making this useful for site-specific cross- linking.
• Carbonyls do not readily exist in proteins; however, mild oxidation of sugar moieties using sodium metaperiodate will convert vicinal hydroxyls to aldehydes or ketones.
• linker molecules may be homobifunctional or heterobifunctional, depending upon the nature of the molecules to be conjugated.
• Homobifunctional cross-linkers have two identical reactive moieties.
• Heterobifunctional cross-linkers are defined as having two different reactive moieties that allow for sequential conjugation reaction.
• Various types of commercially available cross-linkers are reactive with one or more of the following groups: primary amines, secondary amines, sulphydryls, carboxyls, carbonyls and carbohydrates.
• amine-specific cross-linkers are bis(sulfosuccinimidyl) suberate, bis[2-(succinimidooxycarbonyloxy)ethyl] sulfone, disuccinimidyl suberate, disuccinimidyl tartarate, dimethyl adipimate*2 HC1, dimethyl pimelimidate> ⁇ 2 HC1, dimethyl suberimidate> ⁇ 2 HC1, and ethylene glycolbis-[succinimidyl-[succinate]].
• Cross-linkers reactive with sulfhydryl groups include bismaleimidohexane, l,4-di-[3'-(2'-pyridyldithio)-propionamido)] butane, l-[p-azidosalicylamido]-4-[iodoacetamido]butane, and N-[4-(p-azidosalicylamido) butyl]-3'-[2'-pyridyldithio]propionamide.
• Cross-linkers preferentially reactive with carbohydrates include azidobenzoyl hydrazine.
• Cross-linkers preferentially reactive with carboxyl groups include 4-[p-azidosalicylamido]butylamine.
• Heterobifunctional cross-linkers that react with amines and sulfhydryls include N-succinimidyl-3-[2-pyridyldithio]propionate, succinimidyl[4-iodoacetyl]aminobenzoate, succinimidyl 4-[N-maleimidomethyl] cyclohexane-1-carboxylate, m-maleimidobenzoyl-N-hydroxy succinimide ester, sulfosuccinimidyl 6-[3-[2-pyridyldithio]propionamido]hexanoate, and sulfosuccinimidyl 4-[N-maleimidomethyl]cyclohexane-l-carboxylate.
• Heterobifunctional cross-linkers that react with carboxyl and amine groups include l-ethyl-3-[[3-dimethylaminopropyl]- carbodiimide hydrochloride.
• Heterobifunctional cross-linkers that react with carbohydrates and sulfhydryls include 4-[N-maleimidomethyl]-cyclohexane-l-carboxylhydrazide ⁇ 2 HCl, 4-(4-N-maleimidophenyl)-butyric acid hydrazide> ⁇ 2 HCl, and 3-[2-pyridyldithio]propionyl hydrazide.
• the cross-linkers are bis-[B-4-azidosalicylamido)ethyl]disulfide and glutaraldehyde.
• Amine or thiol groups may be added at any nucleotide of a synthetic nucleic acid so as to provide a point of attachment for a bifunctional cross-linker molecule.
• the nucleic acid may be synthesized incorporating conjugation-competent reagents such as Uni-Link AminoModifier, 3'-DMT-C6-Amine-ON CPG, AminoModifier II,
• N-TFA-C6-AminoModifier C6-ThiolModifier, C6-Disulfide Phosphoramidite and C6-Disulfide CPG (Clontech, Palo Alto, CA).
• the agent may be desirable to attach the agent to the linking molecule by a bond that cleaves under normal physiological conditions or that can be caused to cleave specifically upon application of a stimulus such as light, whereby the agent can be released.
• the agent may be inactive in its conjugated form and activated only when released.
• the agent would be released to exert an activity remote from its point of attachment to the body tissue.
• the agent would be released in a sustained fashion, to prolong the release of the agent versus an agent applied to tissue but not covalently coupled to the tissue (or part of a microparticle).
• Readily cleavable bonds include readily hydrolyzable bonds, for example, ester bonds, amide bonds and Schiff s base-type bonds. Bonds which are cleavable by light are well known.
• Noncovalent methods of conjugation may also be used.
• Noncovalent conjugation includes hydrophobic interactions, ionic interactions, high affinity interactions such as biotin-avidin and biotin-streptavidin complexation and other affinity interactions.
• a molecule such as avidin is attached to a linking molecule (which is attached to a reactive moiety). This conjugate, once attached to tissue according to the invention, then becomes a universal linking moiety for any agent attached to a biotin molecule.
• the reactive compounds may be part of a microparticle such as a microsphere or a microcapsule (of micrometer and nanometer size), and the agent may be contained in the microparticle, either physically entrapped therein, covalently bonded thereto or otherwise physiochemically attached to the microparticle.
• a microparticle such as a microsphere or a microcapsule (of micrometer and nanometer size)
• the agent may be contained in the microparticle, either physically entrapped therein, covalently bonded thereto or otherwise physiochemically attached to the microparticle.
• the methods for manufacturing microparticles according to the prior art are well documented and do not form a basis for the present invention. Examples of microspheres, nanospheres, microcapsules and nanocapsules and their method of manufacture may be found in U.S. Patent 5,075,019, PCT WO95/24929, PCT WO94/23738 and PCT WO/97/03657, the disclosures of which are incorporated herein by reference
• the microparticles of the invention possess reactive moieties on their surface (i.e., surface available reactive moieties).
• Reactive moieties include the reactive moieties of Group A, N-hydroxyl-succinimide, and N-alkyl maleimide and derivatives thereof, as well as any of the reactive moieties described herein.
• Microparticles may also possess on their surface reactive compounds (which comprise such reactive moieties). Such reactive compounds include both monofunctional and bifunctional compounds of Formula I, Formula II and Formula III
• the microparticles of the invention contain an agent which when released from the microparticle provides prophylactic, therapeutic or cosmetic benefit to an external body surface with which it is in contact.
• microparticles of the invention are intended for use on a proteinaceous surface including an external body surface such as skin, hair or nails. Microparticles which remain attached to the external surface and which do not degrade substantially throughout the course of treatment (e.g., days or weeks) are most useful in the invention. Any microparticle that contains an agent (as described herein) and that can hold (as a result of the covalent binding described herein) and release the agent onto an external surface (e.g., a skin surface) for a period of time sufficient for the active agent to achieve its prophylactic, therapeutic or cosmetic purpose is useful in the invention. Microparticles commonly effect delivery of agents by way of diffusion, or by degradation or erosion.
• Examples of diffusional systems in which the active agent permeates at a controlled rate from a polymer are described in U.S. Patents 3,854,480, 5,133,974 and 5,407,686.
• Examples of erosional systems in which the active agent is contained within a matrix which in turn erodes with time are described in U.S. Patent 4,452,775, 4,675,189 and 5,736,152.
• the invention provides microparticles which are either biodegradable or non- biodegradable.
• biodegradable refers to the ability of a substance (in this case, a microparticle) to degrade in vivo (e.g., upon contact with external surfaces such as the skin or upon entry into the body).
• biodegradable microparticles are made from polymers having bonds which are easily hydrolyzed once in contact with a physiological environment.
• covalent linkage of the microparticle to the skin, hair or nails is desired. It is the covalent linkage which keeps the microparticles on the skin for the desired time, preferably in a layer, to achieve uniform and extended release of the active agent as desired.
• biodegradable microparticles are embraced by some aspects of the invention.
• the biodegradable microparticles degrade substantially only after the period of time corresponding to the treatment (e.g., days or weeks) in order to ensure sufficient delivery of the active agent to the skin surface.
• Microparticles that are differentially biodegradable are also useful in the invention.
• a "differentially biodegradable" microparticle is one which does not degrade uniformly throughout its volume. It may instead degrade initially in an internal or core region, as an example.
• Internally degradable microparticles may be formed by coating biodegradable cores with non-biodegradable porous films or shells.
• the microparticle may alternatively degrade from the outer surface, however, it would still be necessary that a sufficient amount of reactive moieties remains covalently attached at the surface and extending within the microparticle even throughout the portion of the degradation process during which covalent attachment of the microparticle is desired. This can be achieved, for example, by a microparticle which is covalently crosslinked internally.
• the microparticles are substantially non-biodegradable at their point of attachment to the skin surface over the period of time during which covalent attachment is desired.
• microparticle which is useful to the invention is one which is non- biodegradable.
• a non-biodegradable microparticle is one which does not degrade upon exposure to a physiological environment or temperature. As mentioned above, such non- biodegradable particles release active agent by diffusion.
• the microparticles are substantially non-biodegradable during the treatment period, which may last for several days to several weeks or completely non-biodegradable. In this instance, the microparticles will simply be sloughed off along with the dead skin cells to which they are attached. As is well known to those of ordinary skill in the art, the outermost portion of the skin (to which the microparticles will be attached in some instances) is not living and is _ 26 _
• microparticles having amines or lysines are non-biodegradable.
• the microparticles of the invention may be synthesized using naturally occurring or non-naturally occurring polymers.
• Non-naturally occurring polymers are referred to herein as synthetic polymers.
• Naturally occurring polymers include nucleic acids, peptides, polypeptides, carbohydrates, alginate, polysaccharides (e.g., dextran, cellulose and glycogen), lipopolysaccharides, chitosan, chitin, peptidoglycans, starch, glycosaminoglycans, collagen, rubber (cis-l,4-polyisoprene), guayule (Parthenium argentatum), collagen, chemical derivatives thereof (substitutions, additions of chemical groups, for example, alkyl, alkylene, hydroxylations, oxidations, and other modifications routinely made by those skilled in the art), albumin and other hydrophilic proteins, zein and other prolamines and hydrophobic proteins, copolymers and mixtures thereof.
• the microparticles of the invention may contain natural, synthetic or chimeric polymers provided that, in the formed microparticles, such polymers are their constituent subunits do not saturate, occupy or otherwise interfere with the availability of all reactive moieties required for covalently attaching the microparticles to the body tissue.
• the microparticles may be synthesized and may contain a variety of polymers and other compounds, provided that there still remains an amount of reactive moieties that are surface available sufficient for covalently attaching the microparticles to the body tissue.
• the microparticles may further comprise one or more synthetic polymers or co- polymers.
• synthetic refers to a substance which is not naturally occurring.
• Exemplary synthetic polymers include, but are not limited to, polyamides, polycarbonates, polyalkylenes, polysulfones, poly(2-sulfobutyl-vinyl alcohol)-graft-poly(D,L- lactic-co-glycolic acid), poly-hydroxyalkanoates, polyalkylene glycols, polyalkylene oxides, polyalkylene terepthalates, polydimethylsiloxane polyvinyl alcohols, polyvinyl ethers, polyvinyl esters, polyvinyl halides, silicones, polyglycolic acid (PGA), polylactic acid (PLA), copolymers of lactic and glycolic acids (PLGA), polyanhydrides, polyorthoesters, polyvinylpyrrolidone, polyglycolides, polysiloxanes, polyurethanes and co-polymers thereof, alkyl cellulose, hydroxyalkyl celluloses, cellulose ethers, cellulose esters, nitro
• Still other microparticles may be comprised of chimeric polymers of synthetic and naturally occurring residues.
• Chimeric polymers refer to polymers of different residues or units.
• a chimeric polymer may contain amino acid and non-amino acid residues, or it may contain natural and synthetic residues.
• a residue in a polymer refers to (and may be used interchangeably with) a unit of a polymer.
• Examples of a polymer residue i.e., a polymer unit
• examples of a polymer residue include an amino acid in a peptide and a nucleotide in a nucleic acid.
• Non-amino acid residues such as saccharides, fatty acids, sterols, isoprenoids, purines, pyrimidines, derivatives or structural analogs of the above, or combinations thereof and the like may be used.
• Non-naturally occurring non-amino acid substitutes include but are not limited to 2-azetidinecarboxylic acid, pipecolic acid, S- ethylisothiourea, 2-NH 2 -thiazoline and 2-NH 2 -thiazole.
• the microparticles of the invention may contain natural, synthetic or chimeric polymers provided that such polymers or their constituent subunits do not saturate all reactive moieties required for covalently attaching the microparticles to the body tissue.
• the microparticles may be synthesized using, and may contain, a variety of polymers and other compounds, provided that there still remains an amount of reactive moieties, that are surface available, sufficient for covalently attaching the microparticles to the body tissue.
• the natural, synthetic and chimeric polymers may themselves be biodegradable or non-biodegradable, as intended herein.
• biodegradable polymers include synthetic polymers such as polymers of lactic acid and glycolic acid, polyanhydrides, poly(ortho)esters, polyurethanes, poly(butic acid), poly(valeric acid), and poly(lactide- cocaprolactone), and natural polymers such as those listed herein. In general, these materials degrade either by enzymatic hydrolysis or exposure to water in vivo, by surface or bulk erosion.
• the polymers may optionally be in the form of a hydrogel that can absorb up to about 90% of its weight in water and further, optionally may be crosslinked with multivalent ions or other polymers.
• non-biodegradable synthetic polymers examples include latex, polystyrene, polystyrene derivatives, poly-N-ethyl-4-vinylpyridinium bromide, silicone, polypropylene, ethylene vinyl acetate, poly(meth)acrylic acid, polymethylacrylate, polyamides, copolymers and mixtures thereof.
• U.S. Patent 5,861,149 discloses methods for making non-biodegradable microparticles which can be used in the present invention.
• Polystyrene particles useful in the invention are commercially available from a variety of manufacturers including Polysciences, Inc. (Warrington, PA), Seradyn (Indianapolis, IN) and Dynal.
• the microparticles may also be formed from or may include non-polymer moieties such as lipids including sterols such as cholesterol, cholesterol esters and fatty acids or neutral fats such as mono- di- and tri-glycerides.
• the microparticles may be made from organic and/or inorganic substances.
• the majority of polymers listed above are organic.
• inorganic substances include but are not limited to polyphosphate, zirconia-silica (ZS), Si(OC 2 H 5 ) 4 , Al(NO 3 ) 3 x 9H O, AgNO 3 , HNO 3 , poly(phenylphosphinoborane) (an inorganic analogue of polystyrene) and PRIMM.
• crosslinking agents include chemicals such as glutaraldehyde, dithiobis(succinimidyl) propionate, carbodiimide, and N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP), 1 ,4-Bis(acryloyl)piperazine, N-Hydroxysulfosuccinimide, as well as electromagnetic radiation such as UV radiation.
• SPDP N-succinimidyl-3-(2-pyridyldithio) propionate
• SPDP N-succinimidyl-3-(2-pyridyldithio) propionate
• SPDP N-succinimidyl-3-(2-pyridyldithio) propionate
• SPDP N-succinimidyl-3-(2-pyridyldithio) propionate
• 1 ,4-Bis(acryloyl)piperazine 1,4
• the microparticles may be predominantly composed of one or more polymers.
• a blend of natural and synthetic polymers may be used in microparticle synthesis.
• the microparticles may have an external coating composed of the same or a different polymer or non-polymer substance.
• the microparticle may be composed internally of polystyrene and an active agent and may have an exterior coating (preferably covalently attached) of a substance rich in the reactive moieties useful in the invention.
• more than one polymeric or non-polymeric substance, or a combination thereof may be commingled prior to microparticle formation, resulting in their combined presence both internally and on the external surface of the microparticle.
• the compounds of Formula I, Formula II or Formula III may be incorporated into the microparticles at the time of synthesis or, alternatively, they may be linked to the surface of a pre-formed microparticle.
• the microparticles, once formed, may be coated with a suspension of Formula I, II or III compounds.
• Formula II compounds (and bifunctional reactive compounds of Formula III) may be used together with microparticles by applying these compounds to the body tissue, applying the microparticles to the body tissue, and allowing crosslinking to occur. In this latter embodiment, one reactive moiety of the compound crosslinks to the tissue while the other crosslinks to the microparticle.
• Microparticles having surface available amines or thiols are most preferred in these embodiments.
• the bifunctional reactive compounds may be applied to the body tissue, preferably before, but also simultaneously or following the application of the microparticles.
• the agent is present in the microparticle and is not covalently attached to the reactive moieties of Group A, N-hydroxy succinimide or N-alkyl maleimide.
• the microparticle possesses on its surface these reactive moieties and is capable of covalently attaching to the body tissue.
• the reactive moieties of Group A may be attached to a substituent of the microparticle, such as a polymer, or to a linker.
• microparticle embraces particles, spheres and capsules of both nanometer and micrometer sizes (i.e., microparticles, microspheres, nanoparticles, nanospheres, microcapsules and nanocapsules).
• the microparticles may adopt a variety of shapes including regular shapes such as spheres and ellipses as well as non-regular shapes. Additionally, the surface may be, but need not be, smooth.
• the microparticles may be hollow with the agent stored in the core of the shell, in which case, they may be referred to as microcapsules or nanocapsules.
• porous microparticles are one having internal, potentially interconnected channels (or pores) which are preferably open to the external surface of the particle.
• Methods for synthesizing hollow and porous microparticles are well known in the art. Porous microparticles are generally made by the inclusion of a porogen during microparticle synthesis followed by its removal (e.g., through dissolution in an appropriate WO 01/06829 _ 3Q _ PCT/US00/20210
• the porous microparticles may additionally have a coating comprising reactive moieties, while being internally void of these groups.
• the microparticle be less than five microns in size (i.e., any single dimension of the microparticle is less than 5 microns).
• the microparticle should be small enough so as to feel smooth if applied to the external body surface.
• the microparticles with surface available reactive moieties are small enough to penetrate the external surface, preferably up to but not including the layer of living cells.
• the average size of the microparticle is less than 1 micron but greater than 100 nm, and in others, it is 100 nanometers to 500 nanometers. In still other embodiments, the average size of the microparticles is less than 100 nm in size, 20 nm to 90 nm in size, or 20 nm to 35 nm in size. In most instances, it may be undesirable for the microparticles to penetrate the living layer, and it is therefore intended that the microparticle remain in the cornified layer. It is believed that microparticles less than 1 nm, as well as others that are less than 5 nm could penetrate to the living layer, and thus should be avoided in most instances.
• the particles may enter the cornified layer and thereby release their active agent which may diffuse into and thereby enter the living layer.
• microparticles of an average size of 1 nm to 20 nm, 1 nm to 10 nm and in particular 5 nm to 10 nm may be desirable in some aspects of the invention. It is well within the realm of the ordinary artisan to determine the size of particles which are best suited to the various embodiments recited herein. It also should be noted that size may be relatively uniform as in all the particles being of a certain size, or range or size may be mixed.
• all the microparticles should be at least a certain size (e.g., at least 100 nm). If it is desired that all microparticles penetrate the outer most surface of the epidermis, then they should be no larger than a certain size (e.g., no larger than 100 nm). It also may be desired to have a variety of sizes whereby the microparticles will penetrate to different extents depending on size, thereby forming a three dimensional "layer". Size will depend upon factors such as the agent to be delivered, the condition being treated, the desired length of treatment, and other such factors well known to those of ordinary skill in the art.
• the microparticles are linked to proteinaceous material.
• the microparticles When used in vivo, the microparticles are attached to a body tissue.
• Particularly important body tissues as sites of attachment are the integument (including specifically skin, nails, hair, mucous membranes and the surface of the eye), internal organs, internal tissue and wound beds.
• the tissue In in vitro applications, the tissue may be a body tissue, a tissue or cell isolate, isolated proteins, synthetic proteins, cell cultures and the like for use, for example, in assay systems according to the invention.
• the body tissue is a skin, nail or hair surface.
• the microparticles are water insoluble and, preferably, detergent insoluble (i.e., neither the microparticle nor the bond between the microparticle and the external surface are adversely affected by exposure to detergents, such as hand, body and hair soap).
• detergent insoluble i.e., neither the microparticle nor the bond between the microparticle and the external surface are adversely affected by exposure to detergents, such as hand, body and hair soap.
• Many of the organic polymers listed herein are water insoluble. It is well within the realm of the ordinary artisan to determine which of these are preferred for making water insoluble particles.
• Microparticles may be rendered substantially detergent insoluble by cross-linking or by bonding of noncovalent nature that similarly renders the microparticle insoluble. If cross-linking is used, it is recommended that the reactive moieties of Group A, N-hydroxy succinimide or N-alkyl maleimide are protected to prevent them from participating in the crosslinking, that a crosslinking agent be used which does not involve these reactive moieties or that the reactive moieties be attached to the microparticle after such cross-linking. In another embodiment, the microparticles can be made more resilient to detergent treatment by the incorporation of fluorinated steroids as taught in U.S. Patent 4,927,687.
• the microparticles must possess reactive moieties, such as those of Group A, N-hydroxy-succinimide, N-alkyl-maleimide, or any of the reactive compounds described herein as compounds of Formulae I, II or III.
• the reactive moieties may be provided by any compound which contains them, including, but not limited to, compounds of Formulae I, II and III. As suggested in the foregoing discussion, it is important that these reactive moieties be accessible to the body tissue (e.g., the skin) to which the microparticles are to be bound.
• the reactive moieties must be sufficiently exposed, and the "backbone" to which they are attached preferably sufficiently flexible, to react with and form a covalent bond with reactive molecules on the contacted surface (such as lysines or cysteines).
• reactive molecules on the contacted surface such as lysines or cysteines.
• Reactive moieties which are present on the surface of the microparticles are likely to be accessible, and thus such "surface available" reactive moieties are generally preferred.
• Free surface available reactive moieties may be "free” or "fixed.” Free surface available reactive moieties include those which are present on a free, unconstrained end of a polymeric or non-polymeric substance, present at the surface of the microparticle. The free, unconstrained end of the polymeric or non-polymeric substance may be any length, provided the reactive moieties contained therein are capable of reacting with the body tissue (e.g., the skin). Free reactive moieties also embrace those which are non-complexed. A non- complexed reactive moiety is one which is not in physical association with another moiety to the extent that it is precluded from contacting and being covalently attached to a reactive group on, for example, the skin.
• a reactive moiety may be present in a loop of a polymer the ends of which are both bound to the surface of the microparticle. As long as the loop is long enough and flexible enough to allow the reactive moieties to contact and react with the body tissue (e.g., skin), this type of "fixed reactive moiety" will be useful.
• the surface available reactive moieties must also be present in an amount sufficient to attach covalently the microparticles to the skin.
• microparticles Polymeric and non-polymeric substances from which the microparticles are synthesized may inherently possess the necessary reactive moieties (or molecules), or they may be derivatized either prior to or following microparticle formation to possess such groups.
• the microparticles may be formed of substances lacking reactive moieties and then coated with a substance which contains these moieties.
• the compounds of Formula I, II or II may be interspersed into the microparticle matrix during synthesis so that such compounds are present but not necessarily covalently bonded to the microparticle matrix.
• the reactive moieties may also be linked to the surface of the microparticle after microparticle formation.
• the surface may be prepared or treated to contain amine or thiol moieties, after which it is exposed to an excess of a bifunctional reactive compound such as a Formula II compound having a reactive moiety which reacts with amines or thiols.
• a bifunctional reactive compound such as a Formula II compound having a reactive moiety which reacts with amines or thiols.
• the microparticle is treated so as to have either amines or thiols on its surface, and even more preferably, the bifunctional reactive molecule has one reactive moiety which reacts with amines and another which reacts with thiols.
• the microparticle may be one treated to have thiol groups on its surface, and a bifunctional reactive molecule is one with a Michael acceptor moiety and a carbonyl moiety.
• the Michael acceptor moiety will react with the thiol groups on the microparticle surface, thereby covalently attaching the microparticle to the bifunctional reactive molecule.
• the resulting conjugate may then be applied to a body tissue and covalently attached there using the remaining reactive moiety in the bifunctional compound.
• Polymers or other backbones rich in amines or thiols also can be covalently attached at the surface of the microparticle using homo and heterobifunctional crosslinkers. It further is envisioned that such polymers (or other backbones carrying reactive moieties) can be tethered to the surface of a microparticle by hydrophobic bonding. The manufacture of such microparticles is well within the realm of the ordinary artisan.
• the methods for manufacturing a variety of microparticles according to the prior art are well documented.
• the present invention differs from those of the prior art, in part, in that the polymers or non-polymers of the microparticle themselves contain or are derivatized (including by tethering) to contain reactive moieties such as those present in Group A, N-hydroxy succinimide and N-alkyl maleimide, and the like at the surface, thereby being available for covalent bonding to the body tissue.
• Preferred polymers are polymers bearing multiple reactive moieties, such as those described herein. Additionally, polymers having such reactive moieties spaced at discrete intervals are also useful in the invention. It has been discovered, surprisingly, that the spacing of the reactive moieties can be important to achieving the results of the present invention.
• a further embodiment of the present invention involves microparticles comprising polymers having multiple units, which each bear a reactive moiety such as those described herein.
• the polymer can be a homopolymer or a heteropolymer.
• Polymers having reactive moieties may contain at least three reactive moieties spaced apart from one another at discrete intervals along the backbone of the polymer, separated by one or more backbone atoms.
• the polymer may comprise, or in some instances consist solely of, contiguous reactive moieties, preferably at least 3, at least 4 and at least 5 such contiguous reactive moieties. Polymers of contiguous units, each carrying a reactive molecule, are preferred.
• a polymer rich in units having reactive moieties is a polymer wherein at least 20%> of the units of the polymer carry a reactive molecule, or wherein the polymer includes at least 3, preferably 4 and most preferably 5 separate and discretely spaced by a regular distance units having reactive moieties. In other embodiments, the polymer includes at least 10, at least 15 or at least 20 separate and discretely spaced units having reactive moieties.
• a chain of as few as two units having reactive moieties can be attached to or tethered to an microparticle to render the microparticle capable of attaching to a body tissue.
• the polymers may also contain at least 30%, at least 40%, at least 50% or more of units having reactive moieties, depending upon the embodiment.
• microparticles it may be desirable to vary not only the number of surface available reactive moieties, but it also may be desirable to tether the reactive moieties to the microparticle via a spacer. This can remove, for example, any problems that might arise from steric hindrance, wherein access to the reactive moiety directly on the surface is hindered.
• spacers can be any of a variety of molecules, preferably nonactive, such as straight or even branched carbon chains of C ⁇ -C 30 , saturated or unsaturated, phospholipids, amino acids, and in particular glycine, and the like, naturally occurring or synthetic.
• Additional spacers include alkyl and alkenyl carbonates, carbamates, and carbamides. These are all related and may add polar functionality to the spacers such as the C ⁇ -C 30 previously mentioned.
• the polymers may also have termini (either amino or carboxy) that are predominantly rich in reactive molecules or in units having reactive moieties.
• the termini are located on the surface of the microparticle.
• the terminus may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 or more units at a terminal end of a polymer.
• the reactive moiety rich stretch of the polymer may also be located on a "loop" of a polymer which is present at the surface of a microparticle.
• the reactive moieties of Group A, N-hydroxy-succinimide, N-alkyl-maleimide and the like are capable of reacting with amine groups (such as those present in lysine) and/or thiol groups (such as those present in cysteines). Consequently, the microparticles described herein can also be made from or attached to polymers and non-polymers which contain amines and thiols, as described above. Polymers rich in amines include but are not limited to albumin and polylysine. Microparticles having surface available amines or thiols are preferably used together with bifunctional reactive compounds.
• microparticles having "reactive moieties” such as those of Group A compounds and the like
• microparticles having amine and/or thiol groups particularly those that are surface available.
• Microparticles having amine and/or thiol groups are particularly useful in embodiments in which bifunctional reactive molecules (such as bis-N-hydroxy-succinimide or bis-N-alkyl-maleimide) are applied to the body tissue (e.g., the skin), and microparticles are separately applied to the body tissue.
• One reactive moiety of the bifunctional molecule would attach to the body tissue and the other would attach to the microparticle having surface available amines or thiols.
• the bifunctional molecule could be premixed with the microparticle prior to application to the body tissue (depending upon the choice of surface available reactive moieties and the choice of reactive moieties in the bifunctional compound), it is preferred in some embodiments that each be applied to the body tissue separately.
• the order of application of the bifunctional molecule and the microparticle can be reversed.
• Embodiments in which one reactive moiety of the bifunctional compound reacts only with a microparticle and the other reactive moiety of bifunctional compound reacts only with the body tissue are preferred in some instances.
• Encapsulated microspheres made from poly(lactide-co-glycolide) and poly( ⁇ -CBZ-L- lysine) and subsequently treated so as to expose surface reactive amino groups have been reported previously. (Zheng and Hornsby, 1999, Biotechnol. Prog. 15:763-767)
• the carbobenzoxy (i.e., CBZ) protective groups are removed using either acid hydrolysis or lithium/liquid ammonia reduction, thereby exposing reactive amine groups. Lithium/liquid ammonia reduction is recommended if microsphere are desired, given its less harsh effect of the external surface of the microparticle.
• the lithium treatment was reported to be more effective in producing surface reactive amino _ 36 _
• the lithium treatment may be preferred.
• the active agent may be added during the formation of the microparticles since the lithium treatment reportedly does not create pores in the surface of the particles and thus will not adversely affect the agent.
• the acid hydrolysis method may be preferred, provided the agent is either resistant to the acid treatment or is loaded into the particles following acid treatment.
• a similar strategy may be used to produce non-biodegradable microparticles, by substituting poly(lactide-co-glycolide) with a non-biodegradable polymer such as those disclosed herein.
• a copolymer of lysine and a synthetic polymer such as, for example, poly(lactic acid-co-lysine) may be used alone to form the microparticles followed by mild acid hydrolysis or lithium treatment.
• Such lysine containing copolymers have been manufactured previously. (Barrera, et al., 1993. J. Am. Chem. Soc.
• microparticles may be made using the technique of Zheng and Hornsby but excluding poly-lysine. After being formed, the microparticles may be coated with a solution of poly( ⁇ -CBZ-L-lysine).
• Commercially available microparticles such as those made from polyacrylamide, polyacrylate, polystyrene, or latex (Bio-Rad Laboratories (Richmond, CA), LKB Mather (Stockholm, Sweden)) or those made from natural polymers such as agarose, crosslinked agarose, globulin, and liposomes (Bio-Rad Laboratories (Richmond, CA), Pharmacia (Piscataway, NJ), IBF (France)) can also be coated with CBZ- protected as well as non-protected lysine containing polymer solutions following agent loading. Microcapsule coating methods are known in the art.
• the screening method involves selecting a microparticle and applying it, in an isolated form, to a proteinaceous material such as a body tissue, a body tissue isolate, or more preferably, a polymer rich in cysteine, a polymer rich in lysine or a polymer rich in cysteine and lysine.
• a proteinaceous material such as a body tissue, a body tissue isolate, or more preferably, a polymer rich in cysteine, a polymer rich in lysine or a polymer rich in cysteine and lysine.
• the microparticle is allowed to remain on the material for a time sufficient for forming covalent bonds with the tissue.
• the microparticle may be loaded with a labeling agent such as a fluorescent dye or a fragrance.
• the labeling agent is covalently fixed to the microparticle such that no label escapes from the microparticle. This will ensure that any label detected on the external surface is indicative of a microparticle that is bound to the surface rather than a label which has exited a microparticle which itself was not capable of binding to the surface.
• the surface may be additionally washed with water and/or a detergent and then again tested for the presence of the microparticle.
• the microparticle Prior to contact with the body tissue, the microparticle is loaded with an agent, either physically entrapped therein, covalently bonded thereto or otherwise physiochemically attached to the microparticle.
• the active agent may be incorporated (i.e., "loaded") into the microparticle either at the time of, or after, microparticle formation, depending upon whether the microparticle formation process would be deleterious to the active agent.
• active agent it is meant that the agent, once coupled to a body tissue (such as skin) in vivo or in vitro, either directly or indirectly via a microparticle, has, maintains or can be released to have a desired activity such as a desired physiological, prophylactic, therapeutic or cosmetic activity.
• agents are pharmaceutical agents, sunscreen agents, insecticides, bactericides, fungicides, etc.
• the active agent is not a labeling agent such as a diagnostic agent.
• the agent is not a cosmetic agent.
• the active agent is a non-nucleic acid active agent.
• a non-nucleic acid active agent refers to an active agent which is not a nucleic acid.
• the active agent is a non-protein active agent.
• a non-protein active agent is an active agent which is not a protein (i.e., it is not composed exclusively of peptide linkages of amino acid residues or units).
• the agent is a noncorneocyte, nonlabeling active agent. Specifically excluded in these particular embodiments are corneocyte proteins. In certain embodiments, the agent also is a non-extracellular matrix protein agent. A non-extracellular matrix protein agent is one that is not an extracellular matrix protein. A nonlabeling active agent is one that is not simply a passive label with no function, when applied to a body tissue, other than being a label. Thus, specifically excluded in some embodiments are labeled corneocyte proteins, labeled fibronectin, labeled extracellular matrix proteins, putrescine, dansylcadaverine, 5-(biotinamido)-pentylamine, fluoresceincadaverine and the like.
• agents used according to the invention do not themselves, in their native form, possess reactive moieties. If desired, however, such agents can be modified according to the invention to contain reactive moieties or alternatively and preferably, in some embodiments, they may be tethered or linked to a reactive moiety such as in Formula I.
• they may be modified so as to contain amine or thiol groups, particularly if they are to be used with a bifunctional reactive molecules such as those described herein. This may be accomplished, for example, by adding amine or thiol groups to the agent to form a "modified” agent, or by covalently coupling an amine or thiol group (in the form of lysine or cysteine, for example) to for a "conjugate.”
• the agent may be linked to the natural, synthetic or chimeric polymer or non-polymer. Such linkage may be covalent in nature.
• any linkage between the active agent and another component of the microparticle is characterized by a bond that cleaves under normal physiological conditions or that can be caused to cleave specifically upon application of a stimulus such as light, whereby the agent can be released.
• Readily cleavable bonds include readily hydro lyzable bonds, for example, ester bonds, amide bonds and Schiff s base-type bonds. Bonds which are cleavable by light are well known.
• the agent may be inactive in its conjugated form and activated only when released. In other instances, the agent would be released to exert an activity remote from the point of attachment of the microparticle to the body tissue.
• Noncovalent methods of conjugation may also be used.
• Noncovalent conjugation includes hydrophobic interactions, ionic interactions, high affinity interactions such as biotin-avidin and biotin-streptavidin complexation and other affinity interactions.
• the active agent is free and not linked to another component of the microparticle.
• the release of the active agent from the microparticle is dependent upon the flow of (physiological) fluids into the porous network of the microparticle, the dissolution of the active agent in such fluids and the exit of fluid and agent from the microparticle.
• the agent would be released in a sustained fashion.
• Active agents in an isolated form may also be used according to the invention. "Isolated” as used herein will depend upon the agent employed. In general, isolated means that the material is essentially free of other substances to an extent practical and appropriate for the intended use of the material. In the case of pharmaceuticals and cosmetics, the materials are likely to be substantially pure. In the case of proteins, the proteins are sufficiently pure and sufficiently free from other biological constituents of the host cells from which the proteins are derived so as to be useful in the methods according to the invention. Typically, such active agents will be at least 95% or more pure.
• a native agent is one as it occurs in nature (isolated or synthesized to duplicate a naturally occurring molecule), without modification or conjugation as described herein.
• the body tissue may be, but need not be, pretreated to facilitate the reaction with compounds of Formula I and II.
• Such treatments include washings, abrasive treatments including physical agents such as pumice, silica and oatmeal, enzymes such as papain, bromelins and the like and chemical agents such as alpha hydroxy acids and glycolic acids.
• the main object is to treat the body tissue so as to expose or create reactive cysteines and/or lysines.
• the body tissue may be pretreated by putting down a layer of reactive molecules, such as by applying to the body tissue polymers rich in lysine, cysteine or both lysine and cysteine or applying bifunctional reactive compounds.
• cysteine, lysine, and polymers of cysteines and lysine are described above. As used herein, such terms embrace nonpeptidic multimers of cysteine and lysine whereby amino acid analogs are used to replace these amino acids in the polycysteine or polylysine substrates.
• Some well known classes of peptide mimetics and pseudopeptides are: azabicycloalkane amino acids; thiazabicycloalkane amino acids; oxazabicycloalkane amino acids; diazabicycloalkane amino acids. D-amino acids are an important embodiment.
• some of the reactive moieties including the N-hydroxyl succinimide and maleimide reactive moieties can be used in a bis form to glue two tissues to WO 01/06829 _ 40 _ PCT/USOO/20210
• the reactive molecules need not also carry an agent.
• Such compounds are shown in Formula II:
• Xi and X 2 are reactive moieties selected from Group A or N-hydroxyl succinimide or N-alkyl-maleimide, and the like. In some embodiments, Xi and X are identical.
• Examples of Formula II compounds include but are not limited to bis-N-hydroxyl succinimide and bis-N-alkyl-maleimide.
• Compounds of Formula II can be supplied to the surfaces of two tissues which then are held in contact with one another for a period of time sufficient to permit crosslinking the tissues to one another.
• compounds of Formula I and III can also be used to seal tissues together, provided that Xj and X 2 are both present. In addition to sealing tissue, these latter compounds may also deliver agents to the tissues in the process.
• the invention further provides methods of treating a subject to attach microparticles to a tissue of the subject by contacting a tissue of the subject with a microparticle having surface available reactive moieties and allowing the microparticles to remain in contact with the tissue for a time sufficient to permit a layer of microparticles to covalently attach to the tissue.
• the reactive moieties are present on the surface of the microparticle in an amount sufficient to attach the microparticle to the skin surface.
• the quantity of surface available reactive moieties which is a "sufficient amount" will vary depending upon a number of factors including, but not limited to, the number, quantity and accessibility of corresponding reactive groups on the body tissue, and the size of the microparticle.
• a sufficient amount of surface available reactive moieties can be achieved, for example, by increasing in the microparticles the number of residues which have the reactive moieties, or by increasing in the microparticles (and particularly at the surface) the number of reactive moieties by preventing their chemical reaction with other reactive moieties either intrinsic or extrinsic to the microparticle. Whether the particles have a "sufficient amount" of surface available reactive moieties can be tested as described herein.
• the tissue is an external surface such as skin, nails or hair. In important embodiments, the tissue is a skin surface.
• the method may also involve contacting the skin with a bifunctional reactive compound (e.g., a compound of Formula II) and contacting the skin with the microparticle, and allowing the bifunctional compound to remain in contact with the skin and allowing microparticles to remain in contact with the bifunctional compound for a time sufficient to permit a layer of microparticles to covalently attach to the skin (e.g., via the bifunctional compound).
• a bifunctional reactive compound e.g., a compound of Formula II
• a subject may be a human, non-human primate, cow, horse, pig, sheep, goat, dog, cat, rabbit or rodent. In all embodiments, human subjects are preferred.
• the subject to be treated according to the methods of the invention is one who will benefit from the treatment with the agents, conjugates and microparticles.
• Such treatment can be prophylactic, such as when the microparticles contain a sunscreen agent or a UV filter, or it can be therapeutic, such as when the microparticles contain an anti-fungal agent.
• the subject may be one in need of cosmetic benefit, in which case the microparticles may contain a cosmetic such as a moisturizer or a skin tanning agent.
• contacting refers to a physical interaction between the body tissue, such as for example the skin surface, and the agents, reactive compounds or microparticles (or between the bifunctional reactive compounds and the microparticles), or, alternatively, the suspension in which the agents, compounds or microparticles are provided.
• "contacting” embraces placing the agents, compounds or microparticles in close enough proximity to the skin to allow for their attachment to the skin via the reactive moieties.
• the reactive moieties are preferably those which are surface available.
• the agents, compounds or microparticles may be applied to the skin alone or, alternatively, they may be provided together with a pharmaceutically acceptable carrier.
• microparticles may be applied to the skin along with a bifunctional compound, as described herein.
• agents, compounds or microparticles can be provided in a formulation commonly intended for application to an external surface, such as a lotion, gel, ointment, jelly, cream, shampoo, detergent or spray (e.g., aerosol).
• agents, compounds or microparticles After contacting the agents, compounds or microparticles with, for example, the skin surface, it is necessary to allow them to remain in contact with the skin surface for a time sufficient to permit a layer of agents, compounds or microparticles to covalently attach to the tissue.
• the agents, compounds or microparticles When the agents, compounds or microparticles are contacted with the skin surface they generally will distribute randomly throughout a volume above the skin surface, or if small enough in size, throughout a volume under the outermost layer of skin. This will also be the case should the agents, compounds or microparticles be provided in a topical formulation such as an ointment.
• agents, compounds or microparticles will contact the skin surface initially, however with time, a sufficient number will settle closer to the skin surface until the point where their reactive moieties will react with counterpart active molecules on the skin, resulting in a covalent bond that tethers the agents, compounds or microparticles to the skin. If the agents, compounds or microparticles are small enough, they will distribute randomly below the outermost layer of skin and preferably in proximity to the layer of living skin cells. A "sufficient number of microparticles" is that number required to provide an effective amount of the active agent to the tissue (e.g., the skin surface). In most cases, this will embrace the amount necessary to achieve a uniform distribution of the agent throughout the surface area of the tissue intended to be treated.
• agents, compounds or microparticles whether applied to the tissue in an isolated form or as part of a formulation, are allowed to settle towards the tissue and thereby form a layer.
• a layer of agents, compounds or microparticles is that amount and distribution that is enough to provide distribution of active agent to, for example, the skin in amounts sufficient to achieve the prophylactic, therapeutic or cosmetic purpose of the agent.
• the agents, compounds or microparticles need not be evenly adjacent to one another in the layer, nor must they be in the same plane (as described herein) provided their distribution above, within or below the outermost layer of skin allows the active agent to be distributed sufficiently.
• the active agent is a sunscreen, it is desirable that it be applied uniformly distributed over an entire area of skin in order to provide maximal effect.
• each microparticle containing the sunscreen may not be necessary, however, that for example the microparticles containing the sunscreen be physically touching each other, provided each microparticle is capable of providing sufficient amounts of the agent for a particular surface area. The same is true for the distribution of compounds containing sunscreens.
• the active agent is a cosmetic
• the layer of agents, compounds or microparticles may be a volume of space over the tissue occupied by the agents, compounds or microparticles.
• the agents, compounds or microparticles may be, but need not be, in a planar arrangement.
• a planar arrangement it is meant that the agents, compounds or microparticles are equidistant from the surface of the tissue.
• a non-planar arrangement indicates that the agents, compounds or microparticles are differentially spaced away from the surface of the tissue.
• the distance of the agent, compound or microparticle from the surface of the tissue may depend upon the location of the reactive moieties which have covalently linked to the tissue. If these are located on long pendent chains, the agent, compound or microparticle may not be contacting the tissue surface at all.
• the microparticles may penetrate the skin to the deeper cornified layers but preferably not into the layer of living cells.
• the particles may be located within the cornified layer of the skin.
• the crosslinked microparticles may then become so large that they are unable to exit this layer and are thus retained there.
• the microparticles may all possess only amines or only thiols or a mixture of amines and thiols or alternatively, a mixture of microparticles some of which possess surface available amines and others which possess surface available thiols.
• the surface reactive moieties of the microparticles will dictate the optimal bifunctional molecules to use. As an example, if all the microparticles possess surface available amines, then the preferred bifunctional reactive molecule is one which has two reactive moieties which react with amines. In some embodiments, the microparticles may be those which degrade following the treatment period.
• microparticles are provided to the skin surface as part of a formulation such as those listed above, it is important that the majority of the active agent does not exit (i.e., leach) from the microparticle and into the formulation prior to contact with the skin.
• the active agent is not substantially soluble in the formulation. Instead, the agent will exit the microparticle only upon contact with the skin. This may occur if, for example, the active agent is specifically soluble at higher temperatures (such as at the skin surface), or in the bodily secretions at the skin surface.
• the microparticle may be made from substances which are temperature or environment sensitive, so that contact with the skin but not necessarily with the formulation induces their disintegration and the subsequent release of the active agent. Thermo sensitive polymers in the form of poly(ether-ester) block copolymers are reported by Cha et al., in U.S. Patent 5,702,717.
• the agents, compounds or microparticles may optionally be combined with a pharmaceutically-acceptable carrier to form a pharmaceutical preparation.
• pharmaceutically-acceptable carrier means one or more compatible solid or liquid filler, diluents or encapsulating substances which are suitable for administration into a human.
• carrier denotes an organic or inorganic ingredient, natural or synthetic, with which the active ingredient is combined to facilitate the application.
• the components of the pharmaceutical compositions also are capable of being commingled with the agents of the present invention, and with each other, in a manner such that there is no interaction which would substantially impair the desired pharmaceutical efficacy.
• the pharmaceutical preparations of the invention are applied in pharmaceutically-acceptable amounts and in pharmaceutically-acceptably compositions.
• Such preparations may routinely contain salt, buffering agents, preservatives, compatible carriers, and optionally other therapeutic agents.
• the salts should be pharmaceutically acceptable, but non-pharmaceutically acceptable salts may conveniently be used to prepare pharmaceutically-acceptable salts thereof and are not excluded from the scope of the invention.
• Such pharmacologically and pharmaceutically-acceptable salts include, but are not limited to, those prepared from the following acids: hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, maleic, acetic, salicylic, citric, formic, malonic, succinic, and the like.
• pharmaceutically-acceptable salts can be prepared as alkaline metal or alkaline earth salts, such as sodium, potassium or calcium salts.
• compositions of the invention should be applied under conditions which enhance the interaction between, and covalent attachment of, reactive moieties incorporated into Formula I and II compounds and proteinaceous material. Those conditions will depend upon the mode of delivery, the pH of the body tissue to the compound is applied, as well as the presence of other facilitating molecules which function to enhance such interactions. Determining the most favorable conditions for such interactions is well within the realm of the ordinary artisan.
• the mode of delivery typically will be topical. Other modes of delivery are, nonetheless, appropriate depending on the condition being treated. Aerosols are an example of an appropriate mode of delivery.
• compositions and pharmaceutical preparations may be administered in effective amounts.
• An effective amount in general, means that amount necessary to achieve the purpose for which the active agent is applied. The effective amount will depend upon the mode of administration, the particular condition being treated, the severity of the condition, the needs of the patient, and the desired outcome. It will also depend upon, as discussed above, the stage of the condition, the age and physical condition of the subject, frequency of treatment and mode of treatment, the nature of concurrent therapy, if any, and like factors well known to the medical practitioner. If the active agent is a pharmaceutical agent, then the amount is that amount necessary to delay the onset of, slow the progression of, halt altogether the onset or progression of, or diagnose a particular condition being treated. In the case of a cosmetic agent, the effective amount will be that amount necessary to achieve the desired cosmetic result. In the case of a sunscreen agent, an effective amount will be that amount necessary to achieve suitable protection from the sun as is conventional.
• doses of active compounds of the present invention would be from about 0.01 mg/kg per day to 1000 mg/kg per day. It is expected that doses ranging from 1-500 mg/kg, and preferably doses ranging from 1-100 mg/kg, and even more preferably doses ranging from 1-50 mg/kg, will be suitable.
• a variety of administration routes are available, although topical routes are preferred. The methods of the invention, generally speaking, may be practiced using any mode of administration that is medically acceptable, meaning any mode that produces effective levels of the active compounds without causing clinically unacceptable adverse effects. Such modes of administration include oral, rectal, topical, nasal or interdermal routes.
• the active agents, the microparticles and the formulations in which they are provided can be in solid, semi-solid or liquid form.
• Solid forms include for example, powders, granules and flakes.
• Semi-solid forms include, for example, gels, creams, gelatins and ointments.
• Formulations for topical administration are known to those of ordinary skill in the art and, in most cases, are commercially available from suppliers such as Paddock Laboratories and Gallipot. Information on topically active and inactive agents, and their commercial suppliers is available from various trade manuals, most particularly, Remington's Pharmaceutical Sciences, United States Pharmacopoeia (USP), National Formulary (NF), Merck Index, Physician's Desk Reference (PDR) and Chemical Abstracts.
• the invention also involves kits. In general, the kits contain a compound of Formula
• Kits containing compounds of Formula II may also contain agents of the invention (e.g., those that are amine and/or thiol containing) in a separate container.
• the kit is a package 10 comprising a housing 12 holding a first container 14, a second container 16 and a third container 18.
• a kit may be comprised of a first container which houses a bifunctional reactive compound of the invention (e.g., a compound of Formula II), either alone, or in a pharmaceutically acceptable carrier, or in a topically applied formation.
• the kit may also contain a second container which houses a microparticle composition comprising the agent.
• a third container may also be provided which contains, for example, a linking molecule for preparing the surface of the body tissue for application of the bifunctional reactive compounds and the microparticles.
• the various containers may also contain preservatives, buffers, vehicles, and the like, as is conventional.
• the kit also houses instructions for using the materials according to the invention, particularly for the topical administration of the linking molecules, compounds and microparticles.
• the instructions may be provided separately from the containers (e.g., on a sheet of paper enclosed in the kit) or on one of the containers (e.g., text on the outside surface of a container).
• the agent may be a sunscreen agent.
• sunscreen agents include: p-aminobenzoate analogs such as 2-ethylhexyl-4-dimethylaminobenzoate (Padimate O); p-methoxy-2-ethyl-hexyl-cinnamate (Parsol 1789); oxybenzone (benzophenone-3); ethylhexylsalicylate; diphenylacrylate polyisobutylene; alkyl-$,$-diphenylacrylate and "-cyano-$,$-diphenylacrylate; l-(4-aminophenyl)-2-morpholinylethanone; (l-(4-methoxylphenyl)-3-(4-tert-butyl-phenyl)-propan-l-3-dione; methyl anthranilate; octocrylene; Tretinoin "-hydroxyacid; diphenylacrylate polyisobutylene;
• sunscreen agents include: 3-benzylidene camphor; 4-methylbenzylidene camphor; allantoin PABA benzalphthalide; benzophenone; benzophenone-1 ; benzophenone-10; benzophenone-11 ; benzophenone-12; benzophenone-2; benzophenone-3; benzophenone-4; benzophenone-5; benzophenone-6; benzophenone-7; benzophenone-8; benzophenone-9; benzyl salicylate; benzylidene camphor sulfonic acid; bornelone; bumetrizole; butyl methoxydibenzoylmethane; camphor benzalkonium methosulfate; cinoxate; DEA-methoxycinnamate; diisopropyl methyl cinnamate; dimethyl PABA ethyl cetearyldimonium tosylate; drometrizole; ethyl cinnamate; ethyl
• sunscreen agents include: derivatives of para-amine benzoic acid (PABA); salicylates; cinnamates; benzophenones; camphors;4-aminobenzoic acid; N,N,N-trimethyl-4-(2-oxoborn-3- ylidenemethyl) anilinium methyl sulphate; homosalate (INN); oxybenzone (INN); 2-phenylbenzimidazole-5- sulphonic acid and its potassium, sodium and triethanolamine salts; 3,3'-(l,4-phenylenedimethylene) bis (7,7- dimethyl-2-oxobicyclo-[2.2.1] hept-1-ylmethanesulphonic acid) and its salts; l-(4-tert-butylphenyl)-3-(4- methoxyphenyl) propane- 1,3-dione; alpha-(2-oxoborn-3-ylidene) toluene-4-sulphonic acid and its salts
• 2-phenylbenzimidazole-5-sulfonic acid 2-phenylbenzimidazole-5-sulfonic acid; benzophenone derivatives, such as 2-hydroxy-4-methoxybenzophenone; dibenzoylmethane derivatives, such as 4-tert-butyl-4'-methoxydibenzoylmethane, or ⁇ , ⁇ -diphenylacrylate derivatives, such as 2-ethylhexyl ⁇ -cyano- ⁇ , ⁇ -diphenylacrylate; p-aminobenzoic acid, cinoxate, diethanolamine, p-methoxycinnamate, digalloyl trioleate, dioxybenzone, ethyl 4->bis>hydroxypropyl!aminobenzoate,
• the agent may also be a cosmetic agent.
• cosmetic components include: Vitamin C; Alpha
• -tocopherol (Vitamin E analog); Ammonium lauryl Sulfate; Cocamidopropyl Betaine; Lauramide DEA;
• Cocamide DEA Methyl paraben; Propyl paraben; Butyl paraben; Salicylic acid; Propylene glycol; EDTA; BHT; BHA; TBHQ; DMDM hydantoin; Imidazolidinyl urea; Potassium sorbate; Sodium Benzoate; phenoxyethanol;
• Polysorbate 20 and 80 Sodium laurylether sulfate; Oleyl betaine; Tego betaine; Sorbitol; Glycerin monolaurate;
• the agent may also be a coloring agent for coloring hair or skin.
• a coloring agent is one which is able to change the color of skin, hair or nails. Color change may be effected through for example, a lightening or darkening of skin, hair or nails. Examples of coloring agents for hair include: 1,2,4-benzenetriacetate;
• 2,4-diaminodiphenylamine 2,4-diaminophenol; 2,4-diaminophenol HCl; 2,4-diaminophenoxyethanol HCl;
• 2-amino-6-chloro-4-nitrophenol 2-aminomethyl-p-aminophenol HCl; 2-chloro-5-nitro-n-hydroxyethyl p-phenylenediamine; 2-chloro-6-ethylamino-4-nitrophenol; 2-chloro-p-phenylenediamine;
• 6-nitro-o-toluidine 6-nitro-o-toluidine; acacia catechu; acid black 1; acid black 52; acid blue 1 ; acid blue 3; acid blue 62; acid blue
• N,N-dimethyl-p-phenylenediamine N,N-dimethyl-p-phenylenediamine sulfate
• N,N-dimethyl-n-hydroxyethyl-3-nitro-p-phenylendiamine n-ethyl-3-nitro PABA; n-methoxyethyl-p-phenylenediamine HCl; n-methyl-3-nitro-p-phenylenediamine; n-phenyl-p-phenylenediamine; n-phenyl-p-phenylenediamine HCl; n-phenyl-p-phenylenediamine sulfate; o-aminophenol; p-aminophenol; p-aminophenol HCl; p-aminophenol sulfate; p-methylaminophenol; p-methylaminophenol sulfate; p-phenylenediamine; p-phenylenediamine HCl; p-phenylenediamine sulfate; phenyl methyl pyrazolone; phloro
• coloring agents are cosmetic colorants which include: acid red 195; aluminum stearate; anthocyanins; beta vulgaris; beta vulgaris; bismuth oxychloride; bromocresol green; bromothymol blue; calcium stearate; capsanthin/capsorubin caramel; CI 10006; CI 10020; CI 10316; CI 10316; CI 11680; CI 1 1710; CI 11725; CI 1 1920; CI 12010; CI 12085; CI 12120; CI 12150; CI 12370; CI 12420; CI 12480; CI 12490; CI
• the agent may also be a moisturizing agent.
• a moisturizing agent is an agent which softens and smoothens skin and in some instances hair. Some moisturizing agents are also humectants in that they are able to hold and retain moisture.
• Emollient agents can be moisturing agents. Moisturizing agents can be used soften skin prior to abrasive events such as shaving. In these latter embodiments, the composition of the invention comprising a moisturizing agent can be supplied in a shaving gel or creme.
• moisturizing agents include: proteoglycans and glycosaminoglycans including hyaluronic acid, crosslinked hyaluronic acid, derivatized hyaluronic acid, chondroitin sulfate; mono- and poly-hydroxyl containing chemicals such as glycerin, sorbitol; pyrrolidine carboxylic acid; proteins such as hydrolyzed animal and vegetable protein, collagens, derivatized collagens, elastins; allantoin; polymer skin conditioning agents; polyols such as glycerol; chitosans; derivatized chitosans; and polyglutamine.
• proteoglycans and glycosaminoglycans including hyaluronic acid, crosslinked hyaluronic acid, derivatized hyaluronic acid, chondroitin sulfate; mono- and poly-hydroxyl containing chemicals such as glycerin, sorbitol; pyr
• moisturizing agents include D,L-panthenol, D-panthenol, vitamin A palmitate, vitamin E acetate, methylsilanetriol mannuronate, natural oils such as tallow oil, macadamia nut oil, borage oil, evening primrose oil, kukui nut oil, rice bran oil, tea tree oil, a medium chain fatty acid ester of glycerol, such as glycerol triheptanoate, glyceryl trioctanoate, glycerol trioctanoate, mineral water, silicones, silicone derivatives; allantoin; dipotassium glycyrrhizinate; stearyl glycyrrhizinate; squalane NF; squalane EX; cetyl ester wax; orange roughy oil; hydrogenated phospholipids; hydrocarbon oils and waxes, such as mineral oil, polyethylene and paraffin; triglyceride esters
• Emollients useful in the invention as moisturizers include: acetamidoethoxybutyl trimonium chloride; acetyl trioctyl citrate; acetylated castor oil; acetylated cetyl hydroxyprolinate; acetylated glycol stearate; acetylated hydrogenated cottonseed glyceride; acetylated hydrogenated lanolin; acetylated hydrogenated lard glyceride; acetylated hydrogenated tallow glyceride; acetylated hydrogenated tallow glycerides; acetylated hydrogenated vegetable glyceride; acetylated lanolin; acetylated lanolin alcohol; acetylated lanolin ricinoleate; acetylated lard glyceride; acetylated palm kernel glycerides; acetylated sucrose diste
• C12-15 alkyl benzoate C12-15 alkyl lactate; C12-15 alkyl octanoate; C12-15 pareth-12 oleate; C12-16 alcohols;
• C18-36 acid triglyceride C18-38 alkyl beeswax; C18-70 isoparaffin; C20-40 alkyl behenate; C20-40 isoparaffin; C24-28 alkyl methicone; C30-45 alkyl methicone; C9-11 alcohols; Calendula officinalis; camelina sativa; cananga odorata; candelilla cera; canola; capryl glycol; caprylic/capric/diglyceryl succinate; caprylic/capric/lauric triglyceride; caprylic/capric/linoleic triglyceride; caprylic/capric/myristic/stearic triglyceride; caprylic/capric/stearic triglyceride; caprylic/capric glycerides; caprylic/capric triglyceride; carnauba; carthamus tinctorius; carthamus tinct
• glyceryl stearate citrate glyceryl stearate diacetate; glyceryl stearate lactate; glyceryl triacetyl hydroxystearate; glyceryl triacetyl ricinoleate; glycine soja; glycine soja; glycol butylene glycol montanate; glycol cetearate; glycol dibehenate; glycol dilaurate; glycol dioctanoate; glycol dioleate; glycol distearate; glycol ditallowate; glycol hydroxystearate; glycol oleate; glycol ricinoleate; glycol stearate; glycosaminoglycans; glycosphingolipids; gossypium; helianthus annus; helianthus annuus; heptylundecanol; hexadecyl methicone; hexamethyldisiloxane;
• tetraisononanoate pentaerythrityl tetraisostearate; pentaerythrityl tetralaurate; pentaerythrityl tetramyri state; pentaerythrityl tetraoctanoate; pentaerythrityl tetraoleate; pentaerythrityl tetrapelargonate; pentaerythrityl tetrastearate; pentaerythrityl trioleate; pentahydrosqualene; perfluoropolymethylisopropyl ether; persea gratissima; persea gratissima; petrolatum; petroleum hydrocarbon; phenyl dimethicone; phenyl methicone; phenyl trimethicone; phosphatidylcholine; pimenta acris; piscum iecur; pistacia vera; placental
• PPG-l-ceteth-5 PPG- 10 butanediol
• PPG- 10 cetyl ether phosphate PPG- 10 jojoba acid
• PPG- 10 jojoba alcohol PPG-l-ceteth-5
• PPG- 10 butanediol PPG- 10 cetyl ether phosphate
• PPG- 10 jojoba acid PPG- 10 jojoba alcohol
• PPG-10 methyl glucose ether PPG-10 oleyl ether; PPG-1 1 stearyl ether; PPG-12; PPG-12/SMDI copolymer; PPG-12 butyl ether; PPG-12-PEG-50 lanolin; PPG-12-PEG-65 lanolin oil; PPG-15; PPG-15 stearyl ether;
• PPG- 15 stearyl ether benzoate PPG- 17; PPG- 17 butyl ether; PPG- 17 dioleate; PPG-2 butyl ether; PPG-2 hydrogenated tallowamine; PPG-2 isostearate; PPG-2 lanolin alcohol ether; PPG-2 myristyl ether propionate;
• PPG-5-laureth-5 PPG-50 oleyl ether; PPG-52 butyl ether; PPG-6-deceth-4; PPG-6-deceth-9; PPG-6-laureth-3;
• Humectants useful in the invention as moisturizing agents include: 1 ,2,6-hexanetriol; acetamide MEA; aluminum hydroxide; arachidyl glycol; arginine PCA; butoxypropanol; butylene glycol; butyloctanol; capryl glycol; carboxymethyl chitosan succinamide; chitosan PCA; copper acetyl tyrosinate methylsilanol; copper PCA; copper PCA methylsilanol; cyclomethicone; diglycerin; dimethicone copolyol acetate; dimethicone copolyol adipate; dimethicone copolyol behenate; dimethicone copolyol butyl ether; dimethicone copolyol hydroxystearate; dimethicone copolyol isostearate; dimethicone copolyol laurate; dimethi
• PEG-10 propylene glycol; PEG-100; PEG-12; PEG-135; PEG-14; PEG-150; PEG-16; PEG-18; PEG-180; PEG-2 lactamide; PEG-20; PEG-20 stearate; PEG-200; PEG-240; PEG-25M; PEG-3 stearate; PEG-32; PEG-4; PEG-40; PEG-45M; PEG-6; PEG-60; PEG-75; PEG-8; PEG-8 stearate; PEG-9; PEG-90; placental protein; polydextrose; polyglucuronic acid; polyglycerin-3; polyglyceryl sorbitol; polysilicone-1 ; polysilicone-2; potassium dimethicone copolyol panthenyl phosphate; potassium dimethicone copolyol phosphate; potassium PCA; PPG-20 methyl glucose ether; PPG-20 methyl glucose ether distearate; PPG-38-but
• the agent can also be a depilatory agent.
• a depilatory agent is an agent which removes body hair.
• depilatory agents include: alkali sulphides; alkaline earth sulphides; ammonium thioglycolate; ammonium thiolactate; barium sulfide; calcium sulfide; calcium thioglycolate; ethanolamine thioglycolate; glyceryl thioglycolate; isooctyl thioglycolate; lithium sulfide; magnesium sulfide; magnesium thioglycolate; mercaptopropionic acid; potassium sulfide; potassium thioglycolate; sodium sulfide; sodium thioglycolate; strontium sulfide; strontium thioglycolate; thioglycerin; thioglycollic acid and its salts; thiolactic acid; and zinc s
• a preferred cosmetic agent is any of the known bulking agents which can be added to the hair or nails to provide 'body' and strength.
• Bulking agents are well known to those of ordinary skill in the art.
• Examples of bulking agents generally include cationic surfactant/polymers, fatty alcohols (non-ionic surfactant), waxes or esters, non-ionic polymers (e.g. polyglycols) for thickening, and insoluble silicone.
• the preferred bulking agent is the cationic surfactant, which places a dispersive charge on the hair.
• cationic surfactants include: quaternary ammonium hydroxides, e.g., tetramethylammonium hydroxide, alkyltrimethylammonium hydroxides wherein the alkyl group has from about 8 to 22 carbon atoms, for example octyltrimethylammonium hydroxide, dodecyltrimethy-ammonium hydroxide, hexadecyltrimethylammonium hydroxide, cetyltrimethylammonium hydroxide, octyldimethylbenzylammonium hydroxide, decyldimethyl-benzylammonium hydroxide, stearyldimethylbenzylammonium hydroxide, didodecyldimethylammonium hydroxide, dioctadecyldimethylammonium hydroxide, tallow trimethylammonium hydroxide, cocotrimethylammonium hydroxide, and the corresponding salts thereof, e.g.,
• Additional bulking agents can be solutions of proteins, peptides, and polynucleotides or combinations thereof.
• Particular bulking agents include collagen, keratins, plant structural proteins, silk, fibrin, mucopolysaccharide and elastin.
• Other examples of bulking agents include: polylysine; biotin, panthenol, glycoprotein, and mucopolysaccharide; amodimethicone; acrylates; dimethicone copolymer; di-isobutyl adipate; isododecane; polypropylene glycol, glycerol, disaccharides, urea, dithiothreitol, edta, methyl paraben, propylparaben; polyvinylpyrrolidone and copolymers or derivatives thereof; for example, copolymers with the ethyl or butyl ester of PVA/MA (partially neutralized), copolymers with vinyl a
• Hair conditioning agents are agents which improve the appearance, texture and sheen of hair as well as increasing hair body or suppleness. Usually these compounds facilitate hair styling.
• hair conditioning agents include: Acetamide MEA; Acetamidoethoxybutyl Trimonium Chloride; Acetylated Lanolin; Acetylated Lanolin Alcohol; Acetylmethionyl Methylsilanol Elastinate; Acrylates/Carbamate Copolymer; Alanine; Albumen; Alfalfa (Medicago Sativa) Oil Unsaponifiables; Almondamidopropalkonium Chloride; Almondamidopropyl Betaine; Aluminum Capryloyl Hydrolyzed Collagen; Aluminum Undecylenoyl Collagen Amino Acids; Amino Bispropyl Dimethicone; Aminopropyl Dimethicone; Aminopropyl Laurylglutamine; Ammoni
• Cylcomethicone Cysteine; Cysteine HCl; Cystine; DATEM; DEA-Cocoamphodipropionate; DEA- Cyclocarboxypropyloleate; DEA-Hydrolyzed Lecithin; DEA-Lauraminopropionate; Decyl Betaine; Decyl Mercaptomethylimidazole; Desamido Collagen; Dextran Hydroxy-propyltrimonium Chloride; Diaminopyrimidine Oxide; Dibehenamidopropyldimethylamine Dilinoleate; DibehenylDjarachidyl Dimonium Chloride; Dibehenyldimonium Chloride; Dibehenyldimonium Methosulfate; Dibutyl Lauroyl Glutamide; Di- C12-15 Alkyl Dimonium Chloride; Di-C12-18 Alkyl Dimonium Chloride; Di-C14-18 Alkyl Dimonium Chloride; Di
• Dihydroxyethyl Stearamine Oxide Dihydroxyethyl Stearyl Glycinate; Dihydroxyethyl Tallowamine/IPDI Copolymer; Dihydroxyethyl Tallowamine Oleate; dihydroxyethyl Tallowamine Oxide; dihydroxyethyl Tallow Glycinate; Dihydroxypropyl PEG-5 Linoleammonium Chloride Phosphate; Diisostearamidopropyl Epoxypropylmonium Chloride; Dilaureth-4 Dimonium Chloride; Dilauryl Acetyl Dimonium Chloride; Dilauryldi onium Chloride; Dilinoleamidopropyl Dimethylamine Dimethicone Copolyol Phosphate;
• Antistatic agents can sometimes also be used as hair conditioning agents.
• Antistatic agents are agents reduce static electricity by neutralizing electrical charge on a surface.
• Antistatic agents include: acetamide MEA; acetamidoethoxybutyl trimonium chloride; acetamidopropyl trimonium chloride; acetum; acetylated lanolin; acetylated lanolin alcohol; acetylated lanolin ricinoleate; acetylmethionyl methylsilanol elastinate; acrylamide/sodium acrylate copolymer; acrylamides copolymer; acrylates/ammonium methacrylate copolymer acrylates/pvp copolymer; acrylates copolymer; adipic acid/dimethylaminohydroxypropyl diethylenetriamine copolymer; adipic acid epoxypropyl diethylenetriamine copolymer; alanine; allanto
• hydrolyzed collagen collagen; betaine; bishydroxyethyl dihydroxypropyl stearaminium chloride; butyl ester of ethylene/MA copolymer butyl ester of PVM MA copolymer; C12-15 alkyl salicylate; C12-16 alcohols; C14-20 isoalkylamidopropylethyldimonium ethosulfate; C 18-22 isoalkylamidopropylethyl-dimonium ethosulfate; calcium pantothenate: calcium pantothenate; canolamidopropyl ethyldimonium ethosulfate; capramide DEA; capryl hydroxyethyl imidazoline; capryloyl collagen amino acids; capryloyl hydrolyzed collagen; capryloyl hydrolyzed keratin; capryloyl keratin amino acids; caprylyl hydroxyethyl imidazoline; carpronium chloride; casein; cere
• trimonium chloride octyldodecyltrimonium chloride; oleamide DEA; oleamide MEA; oleamide MIPA; oleamidopropyl betaine; oleamidopropyl dimethylamine; oleamidopropyl dimethylamine glycolate; oleamidopropyl dimethylamine hydrolyzed collagen; oleamidopropyl dimethylamine lactate; oleamidopropyl dimethylamine propionate; oleamidopropyl ethyldimonium ethosulfate; oleamidopropyl hydroxysultaine; oleamidopropyl PG-dimonium chloride; oleamidopropylamine oxide; oleamidopropyldimonium hydroxypropyl hydrolyzed collagen; oleamine; oleamine bishydroxypropyltrimonium chloride; oleamine oxide;
• PEG-5 stearyl ammonium chloride PEG-5 stearyl ammonium lactate; PEG-5 tall oil sterol ether; PEG-5 tallow benzonium chloride; PEG-50 stearamine; PEG-8 palmitoyl methyl diethonium methosulfate; petrolatum;
• zea mays; zinc hydrolyzed collagen.
• cationic and amphoteric fatty acids such as polyquaternium compounds are useful as hair conditioners or fixatives.
• Examples of cationic amino and quaternary ammonium monomers include, for example, vinyl compounds substituted with dialkyl aminoalkyl acrylate, dialkylamino alkylmethacrylate, monoalkylaminoalkyl acrylate, monoalkylaminoalkyl methacrylate, trialkyl methacryloxyalkyl ammonium salt, trialkyl acryloxyalkyl ammonium salt, diallyl quaternary ammonium salts, and vinyl quaternary ammonium monomers having cyclic cationic nitrogen-containing rings such as pyridinium, imidazolium, and quatemized pyrrolidine, e.g., alkyl vinyl imidazolium, alkyl vinyl pyridinium, and alkyl vinyl pyrrolidine salts.
• the alkyl portions of these, monomers are preferably lower alkyls such as the C1-C3 alkyls, more preferably CI and C2 alky Is.
• octylacrylamide/acrylates/butylaminoethyl methacrylate copolymer a polymer of N-tert-octyl acrylamide, methyl methacrylate, hydroxypropyl methacrylate, acrylic acid and t-butyl aminoethyl methacrylate.
• cationic conditioning compounds include quaternary nitrogen derivatives of cellulose ethers, homopolymers of dimethyldiallyl-ammonium chloride, copolymers of acrylamide and dimethyldiallylammonium chloride, homopolymers or copolymers derived from acrylic acid or methacrylic acid containing cationic nitrogen functional groups attached to the polymer via ester or amide linkages, polycondensation products of N,N'-bis-(2,3-epoxypropyl)-piperazine or of piperazine-bis-acrylamide and piperazine, poly-(dimethylbutenylammonium chloride)- ⁇ , ⁇ -bis-(triethanol-ammonium) chloride.
• Other compounds which are useful as hair fixatives include shellac, polyvinylpyrrolidone-ethyl methacrylate-methacrylic acid tarpolymer, vinyl acetate-crotonic acid copolymer, vinyl acetate-crotonic acid-vinyl neodeconate tarpolymer, poly(vinylpyrrolidone-ethylmethacrylate) methacrylic acid copolymer, vinyl methyl ether-maleic anhydride copolymer, octylacrylamide-acrylate-butylaminoethyl-methacrylate copolymer, and poly(vinylpyrrolidone-dimethylaminoethyl-methacrylate) copolymer and derivatives; thioglycollic acid and its salts and esters; potassium or sodium hydroxide; lithium hydroxide; calcium hydroxide; quinine and its salts; resorcinol; l,3-bis(hydroxy
• anti-foaming agents which are useful as bulking agents include: bisphenylhexamethicone; dimethicone; dimethiconol; hexamethyldisiloxane; hexyl alcohol; isopropyl alcohol; petroleum distillates; phenethyl disiloxane; phenyl trimethicone; polysilicone-7; propyl alcohol; silica dimethyl silylate; silica silylate; tetramethyl decynediol; trimethylsiloxysilicate.
• the agent also can be a tissue sealant. Tissue sealants are those used in wound healing to mechanically seal wounds. The use of transglutaminase to covalently attach such materials would add mechanical and adhesive strength to this sealant. Such tissue sealants are composed typically of fibrinogen, collagen, hyaluronic _ 69 _
• acid synthetic peptides and the like. They also can be polyglutamines, polylysines, or polymers of both glutamine and lysine, corneocyte proteins and the like.
• the agents also can be insect repellants.
• a widely used insect repellant is N-N-diethyl-3-methylbenzamide. Pheromones are also useful as insect repellants.
• the agent also may be cultured cells and cultured body tissues used for wound healing, cartilage replacement, comeal replacements and other like surgical procedures.
• the agent can also be a film forming agent.
• a film forming agent is an agent which produces a continuous film on skin, hair or nails upon application. Film forming agents are useful in wound healing or in some cases as hair fixatives. Examples of film forming agents include: acetyl tributyl citrate; acetyl triethyl citrate; acetyl trioctyl citrate; acrylamide/sodium acrylate copolymer; acrylamides/acrylates/DMAPA/methoxy PEG methacrylate copolymer; acrylamides copolymer; acrylamidopropyltrimonium chloride/acrylates copolymer; acrylates/acetoacetoxyethyl methacrylate copolymer; acrylates/acrylamide copolymer; acrylates/ammonium methacrylate copolymer; acrylates/C 10-30 alkyl acrylate crosspolymer; acrylates/diacetoneacrylamide copolymer
• ethylene/calcium acrylate copolymer ethylene/MA copolymer; ethylene/magnesium acrylate copolymer; ethylene/propylene copolymer; ethylene/sodium acrylate copolymer; ethylene/VA copolymer; ethylene/zinc acrylate copolymer; flexible collodion; gellan gum; glyceryl alginate; glyceryl hydrogenated rosinate: glyceryl polyacrylate; glyceryl rosinate; glycosaminoglycans; guar hydroxypropyltrimonium chloride; gutta percha; ydrogenated styrene/butadiene copolymer; hydrogenated styrene/methyl styrene/indene copolymer; ydrolyzed collagen; hydrolyzed elastin; hydrolyzed keratin; hydroxybutyl methylcellulose; hydroxyethyl ethy
• PEG-crosspolymer pentaerythrityl hydrogenated rosinate; pentaerythrityl rosinate; phthalic anhydride/adipic acid/castor oil/neopentyl glycol/PEG-3/trimethylolpropane copolymer; phthalic anhydride/benzoic acid/trimethylolpropane copolymer; phthalic anhydride/butyl benzoic acid/propylene glycol copolymer; phthalic anhydride/glycerin/glycidyl decanoate copolymer; phthalic anhydride/trimellitic anhydride/glycols copolymer; polyacrylamide; polyacrylamidomethylpropane sulfonic acid; polyacrylic acid; polybutylene terephthalate; polychlorotrifluoroethylene; polydimethylaminoethyl methacrylate; polyethylacrylate; polyethylene; polyethylene terephthalate; poly
• Polyquaternium-11 Polyquaternium- 12; Polyquaternium- 13; Polyquaternium- 14; Polyquaternium- 15;
• Polyquaternium-34 Polyquatemium-35; Polyquatemium-36; Polyquatemium-37; Polyquaternium-39;
• PVP/dimethylaminoethylmethacrylate copolymer PVP/eicosene copolymer
• PVP/hexadecene copolymer PVP/hexadecene copolymer
• PVP/VA/itaconic acid copolymer PVP/V A/vinyl propionate copolymer; PVP/va copolymer; rosin acrylate; rosin hydrolyzed collagen; rubber latex; shellac; shellac cera; sodium acrylate/vinyl alcohol copolymer; sodium carrageenan; sodium dvb/acrylates copolymer; sodium polyacrylate starch; sodium polymethacrylate; sodium polystyrene sulfonate; sodium PVM/MA/decadiene crosspolymer; sodium styrene/acrylamide copolymer; sodium styrene/acrylates copolymer; sodium tauride acrylates/acrylic acid/acrylonitrogens copolymer; soluble _ 7 1 _
• the agent can also be an anti-nerve gas agent.
• An anti-nerve gas agent is an agent which counteracts the effects of a nerve gas agent.
• anti-nerve gas agents include: organophosphate hydrolases such as phosphotriesterase; pyridostigmine, physostigmine, eptastigmine, pralidoxime-2-chloride (2-PAM); potassium 2,3-butadion monoximate; potassium permanganate; sodium phenolate or sodium cresolate; chlorinated lime and magnesium oxide; chloramines; bentonite; and a mixture of atropine and PAM.
• the agent can also be a vitamin including vitamin A,vitamin B, vitamin C, vitamin D, vitamin E, and their provitamin counterparts.
• the agent may be a pharmaceutical agent.
• categories of pharmaceutical agents include: analgesic; amino acid; antagonist; anti-acne agent; anti-allergic; anti-asthmatic; antibacterial; anticholinergic; antifungal; antiglaucoma agent; antihistamine; anti-infective; anti-infective, topical; anti-inflammatory; antikeratinizing agent; antimicrobial; antimycotic; antineoplastic, antineutropenic; antiproliferative; antipruritic; antiseborrheic; carbonic anhydrase inhibitor; cholinergic; cholinergic agonist; diagnostic aids; ectoparasiticide; fluorescent agent; glucocorticoid; hair growth stimulant; histamine H2 receptor antagonists; immunizing agent; immunomodulator; immunoregulator; immunostimulant; immunosuppressant; keratolytic; mucosal protective agent; radioactive agents; wound healing agent.
• Analgesic Acetaminophen; Alfentanil Hydrochloride; Aminobenzoate Potassium; Aminobenzoate Sodium; Anidoxime; Anileridine; Anileridine Hydrochloride; Anilopam Hydrochloride; Anirolac; Antipyrine; Aspirin; Benoxaprofen; Benzydamine Hydrochloride; Bicifadine Hydrochloride; Brifentanil Hydrochloride; Bromadoline Maleate; Bromfenac Sodium; Buprenorphine Hydrochloride; Butacetin; Butixirate; Butorphanol; Butorphanol Tartrate; Carbamazepine; Carbaspirin Calcium; Carbiphene Hydrochloride; Carfentanil Citrate; Ciprefadol
• Enadoline Hydrochloride Epirizole; Ergotamine Tartrate; Ethoxazene Hydrochloride; Etofenamate; Eugenol;
• Fenoprofen Fenoprofen
• Fenoprofen Calcium Fentanyl Citrate
• Floctafenine Flufenisal
• Flunixin Flunixin Meglumine
• Ibufenac Indoprofen; Ketazocine; Ketorfanol; Ketorolac Tromethamine; Letimide Hydrochloride; Levomethadyl Acetate; Levomethadyl Acetate Hydrochloride; Levonantradol Hydrochloride; Levorphanol
• Mefenamic Acid Menabitan Hydrochloride; Meperidine Hydrochloride; Meptazinol Hydrochloride; Methadone
• Oxycodone Terephthalate Oxymorphone Hydrochloride
• Pemedolac Pentamorphone
• Pentazocine Pentazocine Hydrochloride; Pentazocine Lactate; Phenazopyridine Hydrochloride; Phenyramidol Hydrochloride; Picenadol Hydrochloride; Pinadoline; Pirfenidone; Piroxicam Olamine; Pravadoline Maleate;
• Zenazocine Mesylate Zomepirac Sodium ; Zucapsaicin.
• Antiacne Adapalene; Erythromycin Salnacedin; Inocoterone Acetate. Antiallergic: Amlexanox; Astemizole; Azelastine Hydrochloride; Eclazolast; Minocromil; Nedocromil;
• Pirquinozol Poisonoak Extract
• Probicromil Calcium Proxicromil
• Repirinast Tetrazolast Meglumine
• Thiazinamium Chloride Tiacrilast; Tiacrilast Sodium; Tiprinast Meglumine; Tixanox.
• Antiasthmatic Ablukast; Ablukast Sodium; Azelastine Hydrochloride; Bunaprolast; Cinalukast; Cromitrile Sodium; Cromolyn Sodium; Enofelast; Isamoxole; Ketotifen Fumarate; Levcromakalim; Lodoxamide Ethyl;
• Tiaramide Hydrochloride Tibenelast Sodium; Tomelukast; Tranilast; Verlukast; Verofylline; Zarirlukast.
• Antibacterial Acedapsone; Acetosulfone Sodium; Alamecin; Alexidine; Amdinocillin; Amdinocillin Pivoxil; Amicycline; Amifloxacin; Amifloxacin Mesylate; Amikacin; Amikacin Sulfate; Aminosalicylic acid;
• Aminosalicylate sodium Amoxicillin; Amphomycin; Ampicillin; Ampicillin Sodium; Apalcillin Sodium;
• Biphenamine Hydrochloride Bispyrithione Magsulfex; Butikacin; Butirosin Sulfate; Capreomycin Sulfate;
• Cefepime Cefepime Hydrochloride; Cefetecol; Cefixime; Cefmenoxime Hydrochloride; Cefmetazole;
• Cefpimizole Cefpimizole Sodium; Cefpiramide; Cefpiramide Sodium; Cefpirome Sulfate; Cefpodoxime Proxetil; Cefprozil; Cefroxadine; Cefsulodin Sodium; Ceftazidime; Ceftibuten; Ceftizoxime Sodium;
• Palmitate Chloramphenicol Pantothenate Complex ; Chloramphenicol Sodium Succinate; Chlorhexidine Phosphanilate; Chloroxylenol; Chlortetracycline Bisulfate; Chlortetracycline Hydrochloride; Cinoxacin;
• Ciprofloxacin Ciprofloxacin Hydrochloride
• Cirolemycin Cirolemycin
• Clarithromycin Clinafloxacin Hydrochloride
• Clofazimine Cloxacillin Benzathine; Cloxacillin Sodium; Cloxyquin; Colistimethate Sodium; Colistin Sulfate;
• Dicloxacillin Sodium Dihydrostreptomycin Sulfate; Dipyrithione; Dirithromycin; Doxycycline; Doxycycline
• Erythromycin Gluceptate Erythromycin Lactobionate; Erythromycin Propionate; Erythromycin Stearate; Ethambutol Hydrochloride; Ethionamide; Fleroxacin; Floxacillin; Fludalanine; Flumequine; Fosfomycin;
• Fosfomycin Tromethamine Fumoxicillin; Furazolium Chloride; Furazolium Tartrate; Fusidate Sodium; Fusidic
• Ibafloxacin Imipenem; Isoconazole; Isepamicin; Isoniazid; Josamycin; Kanamycin Sulfate; Kitasamycin;
• Meclocycline Sulfosalicylate Megalomicin Potassium Phosphate; Mequidox; Meropenem; Methacycline;
• Nifuratrone Nifiirdazil; Nifurimide; Nifurpirinol; Nifurquinazol; Nifurthiazole; Nitrocycline; Nitrofurantoin;
• Penicillin V Benzathine; Penicillin G Potassium; Penicillin G Procaine; Penicillin G Sodium; Penicillin V; Penicillin V
• Sulfadiazine Sulfadiazine Sodium; Sulfadoxine; Sulfalene; Sulfamerazine; Sulfameter; Sulfamethazine; Sulfamethizole; Sulfamethoxazole; Sulfamonomethoxine; Sulfamoxole; Sulfanilate Zinc; Sulfanitran ;
• Temafloxacin Hydrochloride Temocillin; Tetracycline; Tetracycline Hydrochloride ; Tetracycline Phosphate
• Anticholinergic Alverinc Citrate; Anisotropine Methylbromide; Atropine; Atropine Oxide Hydrochloride;
• Elucaine Ethybenztropine
• Eucatropine Hydrochloride Glycopyrrolate
• Heteronium Bromide Homatropine
• Methylsulfate Tiquinamide Hydrochloride; Tofenacin Hydrochloride; Toquizine; Triampyzine Sulfate;
• Antifungal Acrisorcin; Ambruticin; Amphotericin B; Azaconazole; Azaserine; Basifungin; Bifonazole: Biphenamine Hydrochloride ; Bispyrithione Magsulfex ; Butoconazole Nitrate; Calcium Undecylenate;
• Clotrimazole Cuprimyxin; Denofungin ; Dipyrithione; Doconazole; Econazole; Econazole Nitrate;
• Griseofulvin Hamycin; Isoconazole; Itraconazole; Kalafungin; Ketoconazole; Lomofungin; Lydimycin;
• Mepartricin Mepartricin ; Miconazole; Miconazole Nitrate; Monensin ; Monensin Sodium ; Naftifme Hydrochloride;
• Neomycin Undecylenate Nifuratel ; Nifurmerone; Nitralamine Hydrochloride; Nystatin; Octanoic Acid;
• Ticlatone Tioconazole; Tolciclate; Tolindate; Tolnaftate; Triacetin; Triafungin; Undecylenic Acid;
• Antiglaucoma agent Alprenoxime Hydrochloride ; Colforsin; Dapiprazole Hydrochloride ; Dipivefrin Hydrochloride ; Naboctate Hydrochloride ; Pilocarpine; Pirnabine.
• Antihistaminic Acrivastine; Antazoline Phosphate; Astemizole ; Azatadine Maleate; Barmastine;
• Anti-infective Difloxacin Hydrochloride ; Lauryl Isoquinolinium Bromide; Moxalactam Disodium; Omidazole;
• Pentisomicin Sarafloxacin Hydrochloride
• Protease inhibitors of HIV and other retrovirases Integrase Inhibitors of HIV and other retrovirases
• Cefaclor Ceclor
• Acyclovir Zaovirax
• Norfloxacin Noroxin
• Cefoxitin
• Anti-inflammatory Alclofenac; Alclometasone Dipropionate; Algestone Acetonide; Alpha Amylase; Amcinafal; Amcinafide; Amfenac Sodium; Amiprilose Hydrochloride; Anakinra; Anirolac ; Anitrazafen; Apazone;
• Clopirac Cloticasone Propionate; Cormethasone Acetate; Cortodoxone; Deflazacort; Desonide; _ 76 _
• Desoximetasone Desoximetasone; Dexamethasone Dipropionate; Diclofenac Potassium; Diclofenac Sodium; Diflorasone
• Fenclofenac Fenclorac; Fendosal; Fenpipalone; Fentiazac; Flazalone; Fluazacort; Flufenamic Acid; Flumizole; Flunisolide Acetate; Flunixin ; Flunixin Meglumine ; Fluocortin Butyl; Fluorometholone Acetate; Fluquazone;
• Flurbiprofen Fluretofen; Fluticasone Propionate; Furaprofen; Furobufen; Halcinonide; Halobetasol Propionate;
• Lofemizole Hydrochloride Lomoxicam ; Loteprednol Etabonate; Meclofenamate Sodium; Meclofenamic Acid; Meclorisone Dibutyrate; Mefenamic Acid ; Mesalamine; Meseclazone; Methylprednisolone Suleptanate;
• Orgotein Orpanoxin; Oxaprozin; Oxyphenbutazone; Paranyline Hydrochloride; Pentosan Polysulfate Sodium;
• Prednazate Prifelone; Prodolic Acid; Proquazone; Proxazole; Proxazole Citrate ; Rimexolone; Romazarit ; Salcolex ; Salnacedin; Salsalate ; Sanguinarium Chloride ; Seclazone ; Sermetacin; Sudoxicam; Sulindac;
• Tesimide Tetrydamine ; Tiopinac; Tixocortol Pivalate; Tolmetin; Tolmetin Sodium; Triclonide; Triflumidate;
• Antikeratinizing agent Doretinel; Linarotene; Pelretin.
• Antimicrobial Aztreonam; Chlorhexidine Gluconate; Imidurea; Lycetamine; Nibroxane; Pirazmonam Sodium;
• Propionic Acid Pyrithione Sodium; Sanguinarium Chloride ; Tigemonam Dicholine.
• Antimycotic Amorolfine.
• Antineoplastic Acivicin; Aclarubicin; Acodazole Hydrochloride; Acronine; Adozelesin; Aldesleukin;
• Cyclophosphamide Cytarabine ; dacarbazine; Dactinomycin; Daunorabicin Hydrochloride; Decitabine; Dexormaplatin; Dezaguanine; Dezaguanine Mesylate; Diaziquone; Docetaxel; Doxorubicin; Doxorubicin
• Eflornithine Hydrochloride Elsamitrucin; Enloplatin; Enpromate; Epipropidine; Epirubicin Hydrochloride;
• Gemcitabine Hydrochloride Gold Au 198 ; Hydroxyurea; Idarubicin Hydrochloride; Ifosfamide; Ilmofosine;
• Interferon Alfa-2a Interferon Alfa-2b ; Interferon Alfa-nl ; Interferon Alfa-n3; Interferon Beta-I a; Interferon
• Mitocromin Mitogillin; Mitomalcin; Mitomycin; Mitosper; Mitotane; Mitoxantrone Hydrochloride; Mycophenolic Acid; Nocodazole; Nogalamycin; Ormaplatin; Oxisuran; Paclitaxel; Pegaspargase; Peliomycin;
• Pentamustine Peplomycin Sulfate; Perfosfamide; Pipobroman; Piposulfan; Piroxantrone Hydrochloride;
• Plicamycin Plomestane; Porfimer Sodium; Porfiromycin ; Prednimustine; Procarbazine Hydrochloride;
• Puromycin Puromycin Hydrochloride; Pyrazofurin; Riboprine; Rogletimide; Safingol ; Safingol Hydrochloride;
• Tecogalan Sodium Tegafur; Teloxantrone Hydrochloride; Temoporfin; Teniposide; Teroxirone; Testolactone;
• anti-neoplastic compounds include: 20-epi-l,25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine; amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole; andrographolide; angiogenesis inhibitors; antagonist D; antagonist G; antarelix; anti-dorsalizing morphogenetic protein- 1; antiandrogen, prostatic carcinoma; antiestrogen; antineoplaston; antisense oligonucleotides; aphidicolin g
• suramin suramin; swainsonine; synthetic glycosaminogiycans; tallimustine; tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomerase inhibitors; temoporf ⁇ n; temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine; thaliblastine; thalidomide; thiocoraline; thrombopoietin; thrombopoietin mimetic; thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroid stimulating hormone; tin ethyl etiopu ⁇ urin; tirapazamine; titanocene dichloride; topotecan; topsentin; toremifene; totipotent stem cell factor;
• Tricyclic anti-depressant drags e.g., imipramine, desipramine, amitryptyline, clomipramine, trimipramine, doxepin, nortriptyline, protriptyline, amoxapine and maprotiline
• non-tricyclic anti-depressant drugs e.g., sertraline, trazodone and citalopram
• Ca ⁇ antagonists e.g., imipramine, desipramine, amitryptyline, clomipramine, trimipramine, doxepin, nortriptyline, protriptyline, amoxapine and maprotiline
• non-tricyclic anti-depressant drugs e.g., sertraline, trazodone and citalopram
• Ca ⁇ antagonists e.g., sertraline, trazodone and citalopram
• Calmodulin inhibitors e.g., prenylamine, trifluoroperazine and clomipramine
• Amphotericin B Triparanol analogues (e.g., tamoxifen); antiarrhythmic drugs (e.g., quinidine); antihypertensive drugs (e.g., rese ⁇ ine); Thiol depleters (e.g., buthionine and sulfoximine) and Multiple Drag Resistance reducing agents such as Cremaphor EL.
• the compounds of the invention also can be administered with cytokines such as granulocyte colony stimulating factor.
• Antineutropenic Filgrastim; Lenograstim; Molgramostim; Regramostim; Sargramostim.
• Antiproliferative agent Piritrexim Isethionate.
• Antiprotozoal Amodiaquine; Azanidazole; Bamnidazole; Camidazole; Chlortetracycline Bisulfate;
• Antipraritic Cyproheptadine Hydrochloride ; Methdilazine; Methdilazine Hydrochloride; Trimeprazine Tartrate.
• Antipsoriatic Acitretin; Anthralin; Azaribine; Calcipotriene; Cycloheximide; Enazadrem Phosphate; Etretinate;
• Carbonic anhydrase inhibitor Acetazolamide; Acetazolamide Sodium; Dichlo ⁇ henamide; Dorzolamide Hydrochloride; Methazolamide; Sezolamide Hydrochloride.
• Echothiophate Iodide Isoflurophate; Methacholine Chloride; Neostigmine Bromide; Neostigmine Methylsulfate;
• Corticotropin, Repository Corticotropin Zinc Hydroxide; Diatrizoate Meglumine; Diatrizoate Sodium;
• Diatrizoic Acid Diatrizoic Acid; Diphtheria Toxin for Schick Test; Disofenin; Edrophonium Chloride; Ethiodized Oil; Etifenin; WO 01/06829 _ g 0 _ PCT/USOO/20210
• Impromidine Hydrochloride Indigotindisulfonate Sodium; Indocyanine Green; Iobenguane Sulfate I 123;
• Iobenzamic Acid Iocarmate Meglumine; Iocarmic Acid; Iocetamic Acid; Iodamide; Iodamide Megiumine; Iodipamide Meglumine; Iodixanol; Iodoxamate Meglumine; Iodoxamic Acid; Ioglicic Acid; Ioglucol;
• Ioglucomide Ioglycamic Acid; Iogulamide; Iohexol; Iomeprol; Iopamidol; Iopanoic Acid; Iopentol;
• Iophendylate Iprofenin; Iopronic Acid; Ioprocemic Acid; Iopydol; Iopydone; Iosefamic Acid; Ioseric Acid;
• Iothalamic Acid Iotrolan; Iotroxic Acid; Ioversol; Ioxaglate Meglumine; Ioxagiate Sodium; Ioxaglic Acid; Ioxilan; Ioxotrizoic Acid; Ipodate Calcium; Ipodate Sodium; Isosulfan Blue; Leukocyte Typing Serum;
• Glucocorticoid Amcinonide; Beclomethasone Dipropionate; Betamethasone; Betamethasone Acetate;
• Betamethasone Benzoate Betamethasone Dipropionate
• Betamethasone Sodium Phosphate Betamethasone
• Corticotropin Repository; Corticotropin Zinc Hydroxide; Cortisone Acetate; Cortivazol; Descinolone Acetonide; Dexamethasone; Dexamethasone Sodium Phosphate; Diflucortolone; Diflucortolone Pivalate;
• Flucloronide Flumethasone; Flumethasone Pivalate; Flunisolide; Fluocinolone Acetonide; Fluocinonide;
• Fluocortolone Fluocortolone Caproate; Fluorometholone; Fluperolone Acetate; Fluprednisolone;
• Fluprednisolone Valerate Flurandrenolide; Formocortal; Hydrocortisone; Hydrocortisone Acetate;
• Hydrocortisone Buteprate Hydrocortisone Butyrate; Hydrocortisone Sodium Phosphate; Hydrocortisone Sodium Succinate; Hydrocortisone Valerate; Medrysone; Methylprednisolone; Methylprednisolone Acetate;
• Methylprednisolone Sodium Phosphate; Methylprednisolone Sodium Succinate; Nivazol; Paramethasone
• Prednicarbate Prednisolone
• Prednisolone Acetate
• Prednisolone Hemisuccinate Prednisolone Sodium
• Hair growth stimulant Minoxidil .
• Histamine H2 receptor antagonists Ranitidine (Zantac); Famotidine (Pepcid); Cimetidine (Tagamet); Nizatidine
• Immunizing agent Antirabies Serum; Antivenin (Latrodectus mactans); Antivenin (Micraras Fulvius); Antivenin (Crotalidae) Polyvalent; BCG Vaccine; Botulism Antitoxin; Cholera Vaccine; Diphtheria Antitoxin;
• Diphtheria Toxoid Diphtheria Toxoid
• Diphtheria Toxoid Adsorbed Globulin, Immune
• Hepatitis B Immune Globulin Hepatitis B
• Immunomodulator Dimepranol Acedoben; Imiquimod; Interferon Beta- lb; Lisofylline; Mycophenolate Mofetil; Prczatide Copper Acetate.
• Immunoregulator Azarole; Fanetizole Mesylate; Frentizole; Oxamisole Hydrochloride; Ristianol Phosphate; Thymopentin; Tilomisole.
• Immunostimulant Loxoribine ; Teceleukin.
• Immunosuppressant Azathioprine; Azathioprine Sodium; Cyclosporine; Daltroban; Gusperimus Trihydrochloride; Sirolimus; Tacrolimus.
• Mucolytic Acetylcysteine; Carbocysteine; Domiodol.
• Mucosal Protective agents Misoprostol (Cytotec).
• Radioactive agent Fibrinogen 1 125 ; Fludeoxyglucose F 18 ; Fluorodopa F 18 ; Insulin I 125; Insulin I 131 ; Iobenguane I 123; Iodipamide Sodium I 131 ; Iodoantipyrine I 131 ; Iodocholesterol 1 131 ; Iodohippurate Sodium 1 123 ; Iodohippurate Sodium I 125 ; Iodohippurate Sodium I 131 ; Iodopyracet I 125 ; Iodopyracet I 131 ; Iofetamine Hydrochloride I 123 ; Iomethin I 125 ; Iomethin I 131 ; Iothalamate Sodium I 125 ; Iothalamate Sodium I 131 ; Iotyrosine 1 131; Liothyronine I 125; Liothyronine I 131 ; Merisoprol Acetate Hg
• the invention thus may be used, inter alia, to localize drugs to a tissue such as a wound bed or for localized delivery to a tissue, to hold a drug, insect repellant, bactericide fungicide, growth factors, cytokine, and the like at a particular location to prevent the drug from being flushed away to other body sites where it is not needed, to apply bulking agents and other cosmetic agents to the integuments, such as the skin, hair and nails, to hold sunscreen agents at the surface of the skin for longer periods of time, to hold anti-nerve gas enzymes at the surface of the skin whereby nerve gas can be deactivated, to hold or link chemical agents to the skin which can in turn act as binding sites for other agent or . g 2 .

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• 2000
  • 2000-07-24 WO PCT/US2000/020210 patent/WO2001006829A2/en active Application Filing
  • 2000-07-24 EP EP00950655A patent/EP1203141A1/de not_active Withdrawn
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