BIOACTIVE NANOFIBRES FIELD OF THE INVENTION
[0001] The present invention relates to one or more nanofibres comprising an effective amount of one or more active agents, including one or more active agents being, or obtained, from a plant extract, and methods of using the nanofibres, including in delivery systems suitable for use in therapy or prevention of skin diseases or conditions.
BACKGROUND TO THE INVENTION
[0002] The use of electro spinning to produce a nanofibre matrix that contains an active is known. See for example US patent 7,732,427 and US patent application 13/058,033 published as US 2011/0129510 on 2 June 20 . However, there remains a need for active agent delivery systems that are particularly suitable for delivery to mammalian skin.
[0003] In this specification, where reference has been made to external sources of information, including patent specifications and other documents, this is generally for the purpose of providing a context for discussing the features of the present invention. Unless stated otherwise, reference to such sources of information is not to be construed, in any jurisdiction, as an admission that such sources of information are prior art or form part of the common general knowledge in the art.
[0004] It is an object of the present invention to provide a method of producing one or more nanofibres comprising an effective amount of one or more active agents, including one or more active agents being, or obtained, from a plant extract, or which at least provides the public with a useful choice.
[0005] Other objects of the invention may become apparent from the following description which is given by way of example only.
SUMMARY OF THE INVENTION
[0006] In a first aspect of the invention relates to a one or mote nanofibres comprising
an effective amount of one or more active agent being, or obtained, from a plant extract that is associated with or chemically linked to the one or more nanofibres,
and wherein the one or more nanofibres dissolves at a rate of at least O.lmg/min when solubilised in water at 20° C.
[0007] The invention further relates to a one or mote nanofibres comprising
an effective amount of one or more active agent, wherein one or more of the active agents comprises one or more phenol moieties, and wherein the one or more active agents comprising one or more phenol moieties is covalently bound to the one or more nanofibres,
and wherein the one or more nanofibres
i) dissolves at a rate of at least O. mg/ min when solubilised in water at 20° C; or ii) comprises one or more moieties capable of covalently binding to a phenol moiety or moieties; or
iii) is or comprises a polypeptide comprising one or more tyrosine residues; or iv) has a diameter of from about Onm to about OOOnm; or
v) releases substantially all of one or more of the one or more active agents within about 5 minutes when applied to human skin; or
vi) any combination of two or more of (i) to (iv) above.
[0008] Another aspect of the invention relates to a cosmetic composition that comprises an effective amount of one or more active agent being, or obtained, from a plant extract that is associated with or chemically linked to the one or more nanofibres,
and wherein the one or more nanofibres
i) dissolves at a rate of at least O.lmg/ min when solubilised in water at 20° C; or ii) comprises one or more moieties capable of covalently binding to a phenol moiety or moieties; or
iii) is or comprises a polypeptide comprising one or more tyrosine residues; or iv) has a diameter of from about Onm to about OOOnm; or
v) releases substantially all of one or more of the one or more active agents within about 5 minutes when applied to human skin; or
vi) any combination of two or more of (i) to (iv) above.
[0009] Another aspect the invention relates to a method of promoting skin care or repair in a subject, the method comprising applying a cosmetic composition to a subject in need thereof (a) one or more nanofibres formed from a polymer and (b) an effective amount of one or more active agent being, or obtained, from a plant extract that is associated with or chemically linked to the one or more nanofibres,
and wherein the one or more nanofibres
i) dissolves at a rate of at least O.lmg/ min when solubilised in water at 20° C; or ii) comprises one or more moieties capable of covalently binding to a phenol moiety or moieties; or
iii) is or comprises a polypeptide comprising one or more tyrosine residues; or iv) has a diameter of from about Onm to about OOOnm; or
v) releases substantially all of one or more of the one or more active agents within about 5 minutes when applied to human skin; or
vi) any combination of two or more of (i) to (iv) above.
[0010] Another aspect the invention relates to a cosmetic method of promoting skin care or repair, the method comprising applying to a subject in need thereof (a) one or more nanofibres formed from a polymer and (b) an effective amount of one or more active agent being, or obtained, from a plant extract that is associated with or chemically linked to the one or more nanofibres, and wherein the one or more nanofibres
dissolves at a rate of at least O. mg/ min when solubilised in water at 20° C; or comprises one or more moieties capable of covalently binding to a phenol moiety moieties; or
is or comprises a polypeptide comprising one or more tyrosine residues; or has a diameter of from about Onm to about OOOnm; or
releases substantially all of one or more of the one or more active agents within about 5 minutes when applied to human skin; or
any combination of two or more of (i) to (iv) above.
In a first embodiment the one or more active agent is selected from the group comprising
■ an antioxidant active agent,
■ an active agent that lightens skin,
■ an active agent that preserves or improves moisture retention,
• an active agent that preserves or improves skin elasticity, and
• any combination of two or more thereof.
[0012] In a further embodiment the one or more active agent is selected from the group comprising
■ an anti-inflammatory active agent,
■ an active agent that assists or enhances cell proliferation,
■ an active agent that assists or enhances wound healing,
■ an antibacterial active agent, and
■ and combination of two or more thereof.
[0013] In a further embodiment the one or more active agents comprising one or more phenol moieties is or is derived from a plant extract.
[0014] Another aspect the invention relates to a method of promoting skin care or repair in a subject, the method comprising applying to a subject in need thereof (a) one or more nanofibres
formed from a polymer and (b) an effective amount of one or more active agent being, or obtained, from a plant extract that is covalently linked to the one or more nanofibres.
[0015] Another aspect of the invention relates to a method of assisting or enhancing wound healing in a subject, the method comprising applying to a subject in need thereof a nanofibre matrix formed from a polymer and an effective amount of an active agent that assists or enhances wound healing,
and wherein the one or more nanofibres
i) dissolves at a rate of at least O. mg/ min when solubilised in water at 20° C; or ii) comprises one or more moieties capable of covalently binding to a phenol moiety or moieties; or
iii) is or comprises a polypeptide comprising one or more tyrosine residues; or iv) has a diameter of from about 0mm to about 000mm; or
v) releases substantially all of one or more of the one or more active agents within about 5 minutes when applied to human skin; or
vi) any combination of two or more of (i) to (iv) above.
[0016] Another aspect of the invention relates to an antimicrobial method, the method comprising applying to a subject in need thereof a nanofibre matrix formed from a polymer and an effective amount of an antibacterial active agent,
and wherein the one or more nanofibres
i) dissolves at a rate of at least O.lmg/ min when solubilised in water at 20° C; or ii) comprises one or more moieties capable of covalently binding to a phenol moiety or moieties; or
iii) is or comprises a polypeptide comprising one or more tyrosine residues; or iv) has a diameter of from about Onm to about OOOnm; or
v) releases substantially all of one or more of the one or more active agents within about 5 minutes when applied to human skin; or
vi) any combination of two or more of (i) to (iv) above.
[0017] Another aspect of the invention relates to a method of treating or preventing inflammation, the method comprising applying to a subject in need thereof a nanofibre matrix formed from a polymer and an effective amount of an effective amount of an anti-inflammatory active agent,
and wherein the one or more nanofibres
i) dissolves at a rate of at least O.lmg/ min when solubilised in water at 20° C; or ii) comprises one or more moieties capable of covalently binding to a phenol moiety or moieties; or
is or comprises a polypeptide comprising one or more tyrosine residues; or has a diameter of from about 0mm to about 000mm; or
releases substantially all of one or more of the one or more active agents within about 5 minutes when applied to human skin; or
any combination of two or more of (i) to (iv) above.
Another aspect of the invention relates to a method of promoting or enhancing cell proliferation, the method comprising applying to a subject in need thereof a nanofibre matrix formed from a polymer and an effective amount of an active agent that assists or enhances cell proliferation,
and wherein the one or more nanofibres
dissolves at a rate of at least O. mg/ min when solubilised in water at 20° C; or comprises one or more moieties capable of covalently binding to a phenol moiety or moieties; or
is or comprises a polypeptide comprising one or more tyrosine residues; or has a diameter of from about Onm to about OOOnm; or
releases substantially all of one or more of the one or more active agents within about 5 minutes when applied to human skin; or
any combination of two or more of (i) to (iv) above.
Another aspect of the invention relates to a method of preserving or improving skin elasticity, the method comprising applying to a subject in need thereof a nanofibre matrix formed from a polymer and an effective amount of an active agent that preserves or improves moisture retention,
and wherein the one or more nanofibres
dissolves at a rate of at least O.lmg/ min when solubilised in water at 20° C; or comprises one or more moieties capable of covalently binding to a phenol moiety or moieties; or
is or comprises a polypeptide comprising one or more tyrosine residues; or has a diameter of from about Onm to about OOOnm; or
releases substantially all of one or more of the one or more active agents within about 5 minutes when applied to human skin; or
any combination of two or more of (i) to (iv) above.
Another aspect of the invention relates to a method of preserving or improving skin moisture retention, the method comprising applying to a subject in need thereof a nanofibre matrix formed from a polymer and an effective amount of an active agent that preserves or improves moisture retention,
and wherein the one or more nanofibres
dissolves at a rate of at least O. mg/ min when solubilised in water at 20° C; or comprises one or more moieties capable of covalently binding to a phenol moiety moieties; or
is or comprises a polypeptide comprising one or more tyrosine residues; or has a diameter of from about Onm to about OOOnm; or
releases substantially all of one or more of the one or more active agents within about 5 minutes when applied to human skin; or
any combination of two or more of (i) to (iv) above.
Another aspect of the invention relates to a method of skin lightening, the method comprising applying to a subject in need thereof a nanofibre matrix formed from a polymer and an effective amount of an active agent that lightens skin,
and wherein the one or more nanofibres
i) dissolves at a rate of at least O.lmg/ min when solubilised in water at 20° C; or ii) comprises one or more moieties capable of covalently binding to a phenol moiety or moieties; or
iii) is or comprises a polypeptide comprising one or more tyrosine residues; or
iv) has a diameter of from about Onm to about OOOnm; or
v) releases substantially all of one or more of the one or more active agents within about 5 minutes when applied to human skin; or
vi) any combination of two or more of (i) to (iv) above.
[0022] Another aspect of the invention relates to a method of delivering an antioxidant to skin, the method comprising applying to a subject in need thereof a nanofibre matrix formed from a polymer and an effective amount of an antioxidant,
and wherein the one or more nanofibres
i) dissolves at a rate of at least O.lmg/ min when solubilised in water at 20° C; or ii) comprises one or more moieties capable of covalently binding to a phenol moiety or moieties; or
iii) is or comprises a polypeptide comprising one or more tyrosine residues; or iv) has a diameter of from about 0mm to about 000mm; or
v) releases substantially all of one or more of the one or more active agents within about 5 minutes when applied to human skin; or
vi) any combination of two or more of (i) to (iv) above.
[0023] Use of one or mote nanofibres in the preparation of a medicament that assists or enhances wound healing, that assists or enhances cell proliferation, preserves or improves moisture
retention, preserves or improves skin elasticity, lightens skin, provides an antibacterial effect, provides an anti-inflammatory effect or provides an antioxidant effect, and wherein the one or more nanofibres
i) dissolves at a rate of at least O. mg/ min when solubilised in water at 20° C; or ii) comprises one or more moieties capable of covalently binding to a phenol moiety or moieties; or
iii) is or comprises a polypeptide comprising one or more tyrosine residues; or iv) has a diameter of from about Onm to about OOOnm; or
v) releases substantially all of one or more of the one or more active agents within
about 5 minutes when applied to human skin; or
vi) any combination of two or more of (i) to (iv) above.
[0024] One or mote nanofibres for assisting or enhancing wound healing, assists or enhancing cell proliferation, preserving or improving moisture retention, preserving or improving skin elasticity, lightening skin, providing an antibacterial effect, providing an anti-inflammatory effect or providing an antioxidant effect, and wherein the one or more nanofibres
i) dissolves at a rate of at least O.lmg/ min when solubilised in water at 20° C; or ii) comprises one or more moieties capable of covalently binding to a phenol moiety or moieties; or
iii) is or comprises a polypeptide comprising one or more tyrosine residues; or iv) has a diameter of from about lOnm to about lOOOnm; or
v) releases substantially all of one or more of the one or more active agents within
about 5 minutes when applied to human skin; or
vi) any combination of two or more of (i) to (iv) above.
[0025] Another aspect of the invention relates to a dressing that comprises
a support that carries one or more nanofibres,
an effective amount of one or more active agents that is associated with or chemically linked to the one or more nanofibres,
where the one or more nanofibres dissolves at a rate of at least O.lmg/ min when solubilised in water at 20° C.
[0026] In one aspect the invention relates to a method of forming one or more nanofibres comprising
providing an active agent obtained from a plant extract,
providing cold water fish collagen,
mixing the active agent and the cold water fish collagen in a solvent phase,
electro spinning the mixture of active agent and the cold water fish collagen to form one or more nanofibres associated with the active agent.
[0027] In one aspect the invention relates to one or mote nanofibres formed from cold water fish collagen and additionally comprising about 10 to about 80% by weight of an active agent obtained from a plant extract, and wherein the diameter of the fibres is between about 0 to about OOOnm .
[0028] In one aspect the invention relates to one or mote nanofibres formed from cold water fish collagen and additionally comprising about 10 to about 80% by weight of an active agent obtained from a plant extract, and wherein the rate of solubility of the nanofibres in water at 20° C is at least O.lmg/min.
[0029] The following embodiments may relate to any of the above aspects.
[0030] In various embodiments the polymer is selected from the group comprising
thermoplastics, including PLA, PLGA, PVOH, PVA, PVB, PET, PBT, and PA6/PA6,6;
polypeptides, including collagen and gelatin; polysaccharides, including cellulose and its derivatives (cellulose acetate, carboxymethylated-cellulose (CMC), alginate, chitosan, carrageenan, and xanthan gum; and bacterially-produced plastics, including PHA, PHB, PHBV, and the like.
[0031] In one embodiment the polymer is selected from the group consisting of cold water fish collagen, PVOH, PA-6 and PVB.
[0032] In some embodiments nanofibres are formed into a matrix of nanofibres.
[0033] In some embodiments the nanofibre, nanofibres or matrix of nanofibres are formed onto a support.
[0034] In some embodiments the support is formed from woven material such as cloth, or non-woven material such as plastic.
[0035] In some embodiments the diameter of the nanofibre is about 10, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000 nm.
[0036] In certain embodiment the one or more active agents is present in the nanofibre or nanofibres (i.e., the loading of the active agent in the nanofibre or nanofibres) is about 0.1, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 10, 15, 20, 25 or 30 g/m2.
[0037] In some embodiment the one or more active agents comprises about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75 or 80% by weight of the nanofibre.
[0038] In some embodiments, the one or more active agents is present in the nanofibre matrix at about 0.1, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 10, 15, 20, 25 or 30 g/m2.
[0039] In some embodiment the one or more active agents comprises about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75 or 80% by weight of the nanofibre matrix.
[0040] It will be appreciated that the one or more active agents is typically present in the nanofibre or nanofibres or in the nanofibre matrix at a concentration or amount sufficient to provide an effective dose to the surface to which the nanofibre or nanofibres or nanofibre matrix is applied. For example, in certain embodiments the one or more active agents is present in the nanofibre or nanofibres or in the nanofibre matrix at a concentration or amount sufficient to provide a therapeutic dose to the surface to which the nanofibre or nanofibres or nanofibre matrix is applied, for example is present at a concentration or amount sufficient to deliver about 0.1, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 10, 15, 20, 25 or 30 g per m2 of the surface to which the nanofibre, nanofibres or nanofibre matrix is applied.
[0041] In some embodiments the active agent contains a phenolic moiety. Preferably the active agent is selected from one or more benzoic acids, such as gallic acid, syringic acid, vanillic acid, one or more hydroxycinnamic acids, including caffeic acid, caftaric acid, GRP, p-coumaric acid, coutaric acid, one or more flavan-3-ols, such as catechin, epicatechin, one or more stilbenes, including t- resveratrol, and t-piceid, one or more flavanols, including quercetin, myricetin and rutin, and one or more anthocyanins, including delphinidin, petunidin, cyanidin, malvidin and peonidin.
[0042] In certain embodiments the active agent is a plant enzyme, such as actinidin, papain, or bromelain.
[0043] In some embodiments the association of one or more of the active agents with the one or more nanofibres is by adsorption.
[0044] In some embodiments one or more or the active agents, such as one or more of the active agents comprising one or more phenolic moieties, is covalently bound to the one or more nanofibres (i.e. the chemical linkage is a covalent bond).
[0045] In some embodiments the nanofibre matrix is applied to a support and used as a band aid, bandage, dressing, operating gown, facial mask, facial strip, cosmetic patch or the like.
[0046] In some embodiments the polymer is formed from a single type of monomeric unit (i.e. homop olymer) .
[0047] In some embodiments the polymer comprises two or more different monomer units (i.e. heteropolymer).
[0048] In some embodiments the nanofibre or nanofibres are formed from a single type of polymer (i.e. a homogenous nanofibre).
[0049] In some embodiments the nanofibre or nanofibres are formed from multiple different type of polymer (i.e. a heterogenous nanofibre).
[0050] In some embodiments a single type of nanofibre forms a homogenous matrix of nanofibres.
[0051] In some embodiments multiple types of nanofibres form a heterogeneous matrix of nanofibres.
[0052] In some embodiments where multiple different polymers, nanofibres or matrices are used, whether as single or multiple layers, the different polymers, nanofibres or matrices dissolve or release the active agent at different rates.
[0053] In some embodiments the different polymers are each individually associated with or linked to an active agent, whereas the active agent may be the same or different.
[0054] In some embodiments the additional polymer is a thermoplastic polymer or a biopolymer. In one exemplary embodiment the thermoplastic polymer is selected from the group consisting of PVOH, PA-6, PA-66, PET, PLA, nylon and PVB. In one exemplary embodiment the biopolymer is selected from the group consisting of cellulose, cellulose acetate, gelatine, chitosan, zein, and collagen.
[0055] In some embodiments the matrix of nanofibres is multilayed with each layer being formed from a different polymer or nanofibre or mixture of nanofibres.
[0056] In some embodiments each matrix layer has a different bioactive or mix of bioactives associated or chemically linked to the nanofibre or mixture of nanofibres to that of one or more of the other layer or layers.
[0057] In some embodiments the solvent phase assists the formation of the nanofibre matrix.
[0058] This invention may also be said broadly to consist in the parts, elements and features referred to or indicated in the specification of the application, individually or collectively, and any or all combinations of any two or more of said parts, elements or features, and where specific integers are mentioned herein which have known equivalents in the art to which this invention relates, such known equivalents are deemed to be incorporated herein as if individually set forth
[0059] It is intended that reference to a range of numbers disclosed herein (for example, 1 to 0) also incorporates reference to all rational numbers within that range (for example, 1, 1.1, 2, 3, 3.9, 4, 5, 6, 6.5, 7, 8, 9 and 10) and also any range of rational numbers within that range (for example, 2 to 8, 1.5 to 5.5 and 3.1 to 4.7) and, therefore, all sub-ranges of all ranges expressly disclosed herein are hereby expressly disclosed. These are only examples of what is specifically intended and all possible combinations of numerical values between the lowest value and the highest value enumerated are to be considered to be expressly stated in this application in a similar manner.
[0060] In this specification where reference has been made to patent specifications, other external documents, or other sources of information, this is generally for the purpose of providing a context for discussing the features of the invention. Unless specifically stated otherwise, reference to such external documents is not to be construed as an admission that such documents, or such sources of information, in any jurisdiction, are prior art, or form part of the common general knowledge in the art.
[0061] Other aspects of the invention may become apparent from the following description which is given by way of example only.
DESCRIPTION OF THE FIGURES
[0062] Figure 1 is a graph showing the release of bioactives from Vinanza® skin performance plus extract incorporated into PVOH and PA-6 nanofibres.
[0063] Figure 2 is a graph showing the release of individual phenolic active agents 3-pCoQA (♦), catechin (■), procyanidinB2 (A), and epi-catechin (X) from the PVOH nano fibre matrix as described in Example 3.
[0064] Figure 3 is a graph showing the release of individual phenolic active agents 3-CQA (♦), 3-pCoQA (■), catechin (A), procyanidinB2 (X), and epi-catechin (*) from PA-6 matrix as described in Example 3.
[0065] Figure 5 is a graph showing the effect of Vinanza skin performance plus blend applied as a cream on skin performance.
[0066] Figure 3 is a graph showing the effect of Vinanza skin repair plus blend applied as a PVOH nano-fibre patch on damaged skin.
DETAILED DESCRIPTION OF THE INVENTION
[0067] The invention relates to a one or more nanofibres comprising
an effective amount of one or more active agent, for example one or more active agents being, or obtained, from a plant extract that is associated with or chemically linked to the one or more nanofibres, the active agent selected from the group comprising
■ an active agent that assists or enhances wound healing,
■ an antibacterial active agent,
■ an anti-inflammatory active agent,
■ an active agent that assists or enhances cell proliferation,
■ an antioxidant active agent,
■ an active agent that lightens skin,
■ an active agent that preserves or improves moisture retention, and
■ an active agent that preserves or improves skin elasticity, and
■ and combination of two or more thereof,
and wherein the one or more nanofibres i) dissolves at a rate of at least O. mg/ min when solubilised in water at 20° C; or ii) comprises one or more moieties capable of covalently binding to a phenol moiety or moieties; or
iii) is or comprises a polypeptide comprising one or more tyrosine residues; or iv) has a diameter of from about Onm to about OOOnm; or
v) releases substantially all of one or more of the one or more active agents within about 5 minutes when applied to human skin; or
vi) any combination of two or more of (i) to (iv) above.
[0068] The invention also relates to methods of providing a beneficial effect on the skin, whether that be to the epidermis, dermis or subcutis layers of skin, or any combination thereof.
[0069] The invention also relates to a method of promoting skin care or repair in a subject, the method comprising applying to a subject in need thereof (a) one or more nanofibres formed from a polymer and (b) an effective amount of one or more active agent being, or obtained, from a plant extract that is covalendy linked to the one or more nanofibres.
1. Definitions
[0070] The term "one or more active agent obtained from a plant extract" include analogues of active agents that derive from plant extracts.
[0071] The term "analogues of active agents that derive from plant extracts" and its grammatical equivalents and derivates means an active agent having a structure, function, and/ or composition equivalent to that of an agent obtained from a plant extract. Exemplary methods to determine functional equivalence of active agents, such as those obtained from plants and their analogues, are well known in the art and representative methods are provided herein in the
Examples.
[0072] The term "comprising" as used in this specification means "consisting at least in part of. When interpreting statements in this specification which include that term, the features, prefaced by that term in each statement or claim, all need to be present but other features can also be present. Related terms such as "comprise" and "comprised" are to be interpreted in the same manner.
[0073] As used herein the term "dressing" and its derivates includes patches, strips, bandages or band aids.
[0074] As used herein the term "promoting" and its derivates includes initiating, enhancing or mediating, for example promoting a biological response includes initiating, enhancing or mediating a biological response.
2. Polymer
[0075] The polymer used to form the one or more nanofibres can be any suitable polymer that is compatible with human skin. For example, the polymer can be a thermoplastic such as PVOH, PA-6, PVB, PA-66, PET, PLA, nylon or a mixture thereof, or a bio-polymer such as collagen, cellulose, cellulose acetate, gelatine, chitosan, zein, or a mixture of a thermoplastic and a biopolymer.
[0076] It should be appreciated that any suitable solvent can be used providing it does not leave a toxic residue, solubilises both the polymer and the active agent, does not inactive the active agent, and evaporates off during electro spinning.
[0077] In some embodiments of the invention the biopolymer is collagen sourced from cold water fish, such as hoki. This collagen has certain advantageous properties such as a lower gelling temperature.
[0078] Without wishing to be bound by any theory, the applicants believe that polypeptides, such as collagen, and in particular cold water fish collagens such as hoki collagen, comprising relatively high concentrations of amino acid residues capable of covalendy-linking under conducive conditions with one or more phenolic moieties, are particularly suited to application in the present invention. Exemplary amino acid residues as above include tyrosine, phenylalanine, and tryptophan.
[0079] In one embodiment the polymer may comprise one or more monomeric units. For example, if the polymer is formed from the same monomeric units then it is a homopolymer. In some embodiments the polymer may be a heteropolymer. That is, it is formed from at least two different monomeric units.
[0080] In some embodiments the nanofibre can be formed as a homogenous nanofibre or as a heterogeneous nanofibre. A homogenous nanofibre is one in which the nanofibre is formed from a single polymer, e.g. collagen. A heterogeneous nanofibre is one in which the nanofibre is formed from two or more polymers, for example collagen polymers and PVOH polymers.
[0081] When formed as a multi-polymer nanofibre, fibres formed from one polymer may associate together, for example using van der waals forces, and the second polymer may be associated with the first polymer. For example, in one case of a nanofibre formed from collagen and PVOH, the collagen forms a fibre where the collagen are associated with each other by van der waals forces and the PVOH forms an associated polymer.
[0082] In some embodiments the nanofibres can form a homogenous or heterogeneous matrix of nanofibres. A homogenous matrix of nanofibres is one in which the matrix is formed from a single nanofibre. A heterogeneous matrix of nanofibres is one in which the matrix is formed from multiple different nanofibres.
[0083] In some embodiments the matrix can be a multi-layered matrix. It should be appreciated that the multilayered matrix can be multiple layers of different polymers or nanofibres, or multiple layers of the same polymer or nanofibres. It should also be appreciated that each layer can have embedded or attached a different active agent to the other layer (s). For example, a multilayered matrix formed from nanofibres formed from PVOH may have associated with or attached thereto a first active agent, and a different layer of the matrix, for example formed from PA-6, can have associated with or attached a second and different active agent.
[0084] In certain exemplary embodiments, when formed as a multi-layered matrix, nanofibres formed from one polymer are associated together, for example using van der waals forces, to form a layer, and nanofibres formed from a different polymer are associated together to form another layer. Each layer is then associated with the other, for example by van der waals forces. For example, in
one case of a multilayer nanofibre matrix formed from collagen and PVOH, the collagen nanofibres form a layer where the collagen nanofibres are associated with each other, for example by van der waals forces, and the PVOH nanofibres form an associated layer.
[0085] In some embodiments the diameter of the nanofibre is about 10, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000 nm and useful ranges may be selected between any of these values (for example, from about 10 to about 1000, about 10 to about 900, about 0 to about 750, about 10 to about 600, about 0 to about 500, about 10 to about 300, about 0 to about 00, about 100 to about 000, about 00 to about 950, about 00 to about 800, about 00 to about 600, about 00 to about 500, about 00 to about 350, about 250 to about 1000, about 250 to about 900, about 250 to about 850, about 250 to about 750, about 250 to about 550, about 250 to about 500, about 250 to about 400, about 300 to about 1000, about 300 to about 750, about 300 to about 600, about 300 to about 400, about 400 to about 1000, about 400 to about 900, about 400 to about 700, about 400 to about 600, about 400 to about 500, about 500 to about 1000, about 500 to about 750, about 500 to about 600, about 650 to about 1000, about 650 to about 900, about 650 to about 750, about 700 to about 1000 or about 800 to about 1000 nm).
[0086] In certain exemplary embodiments, the invention provides a multi-layered nanofibre matrix, wherein one or more of the nanofibres comprises an effective amount of one or more active agents as described herein, and wherein at least one of the nanofibres comprising the matrix i) dissolves at a rate of at least O.lmg/ min when solubilised in water at 20° C; or ii) releases substantially all of one or more of the one or more active agents to which it is bound or with which it is associated within about 5 minutes when applied to human skin.
[0087] In various exemplary embodiments, at least one of the nanofibres comprising the matrix releases substantially all of one or more of the one or more active agents to which it is bound or with which it is associated within about 4 minutes, within about 3 minutes, within about 2 minutes, within about 110 s, within about 100 s, or within about 60 seconds, when applied to human skin.
[0088] In certain exemplary embodiments, the invention provides a multi-layered nanofibre matrix, wherein one or more of the nanofibres comprises an effective amount of one or more active agents as described herein, and wherein at least one of the nanofibres comprising the matrix i) dissolves at a rate of at least O.lmg/ min when solubilised in water at 20° C; or ii) releases substantially all of one or more of the one or more active agents to which it is bound or with which it is associated within about 5 minutes when applied to human skin,
and wherein at least one of the nanofibres comprising the matrix
i) is essentially insoluble or is sparingly soluble in water at 20° C; or
ii) releases less than about 1% to about 75% of one or more of the one or more active agents to which it is bound or with which it is associated within about 60 minutes when applied to human skin.
[0089] In various exemplary embodiments, at least one of the nanofibres comprising the matrix releases less than about 50% of one or more of the one or more active agents to which it is bound or with which it is associated within about 00 minutes, within about 200 minutes, within about 300 minutes, within about 400 minutes, within about 500 minutes, within about 600 minutes, within about 700 minutes, within about 800 minutes, within about 900 minutes within about 000 minutes, within about 8 hours, within about 20 hours, within about 22 hours, within about 24 hours, or within about 2 days, when applied to human skin.
[0090] In certain embodiments, at least one of the nanofibres comprising the matrix releases less than about 50% of one or more of the one or more active agents to which it is bound or with which it is associated within about 2 days, within about 3 days, within about 4 days, within about 5 days, or within about 1 week, when applied to human skin.
[0091] It will be appreciated that the targeted selection of active ingredient and polymer enables the production of a multilayer nanofibre matrix having multiple kinetics of active agent release, for example wherein one or more active agents is rapidly released from one polymer or nanofibre present in the matrix, such as to give an antibacterial or antimicrobial effect, and one or more active agents is released over a longer period, for example to support skin repair, recruit immune cells to the site of application, or the like.
3. Active agent
[0092] In certain embodiments, the active agent is a plant extract. This includes bioactives or chemicals isolated from plants, and their analogues. In certain embodiments, the active agent comprises one or more phenolic moiety. It will be appreciated that expressly considered active agents include agents comprising one or more phenolic moieties being or derived from a plant extract.
[0093] Plants contain a range of bioactive compounds, for example, compounds that protect the plants from environmental stress such as UV radiation and microbial contamination.
[0094] Examples of sources of useful bioactives include but are not limited to, grape seed, kiwifruit, blackcurrant, boysenberries and red grape. Further examples include manuka, manuka honey, aloe, ginger, olive or olive leaf, ginseng and the like. Still further examples, seaweed extracts, proteins from plant or animal sources and conjugates of these with plant extracts, essential oils and
extracts of the bioactives from these oils, and extracts of the bioactives from the honey, herbal extracts.
[0095] By way of example, bioactives having antioxidant activity can be obtained from one or more of grapes, boysenberries, blackcurrants and kiwifruit. Bioactives having anti-inflammatory and antioxidant activity can be obtained from grapes, particularly New Zealand grown grapes.
Bioactives having antioxidant and anti-bacterial activity can be obtained from grape and
blackcurrant.
[0096] In some embodiments the bioactives are extracted from the plants using water extraction.
[0097] In some embodiment the loading of the active agent on the nanofibre or nanofibres is about 0.1, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 10, 15, 20, 25 or 30 g/m2.
[0098] In some embodiment the active agent comprises about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75 or 80% by weight of the nanofibre matrix.
[0099] In some embodiments the active agent is chemically bound to the one or more nanofibres. Preferably the active agent forms a covalent bound with the one or more nanofibres.
4. Method of formation of nanofibres
[00100] In one aspect the invention relates to a method of forming a one or more nanofibres comprising
providing an active agent,
providing a polymer or mixture of polymers,
mixing the active agent and the polymer or polymers in a solvent phase,
electro spinning the mixture of active agent and the polymer or polymers to form active imbibed nanofibre or nanofibres.
[00101] In some embodiments the solvent assists the formation of the one or more nanofibres.
[00102] In some embodiments a single polymer is used. For example, collagen may be used to produce a homogenous nanofibre. Alternately, other biopolymers or thermoplastic polymers may be incorporated into the solvent. Examples of suitable thermoplastic polymers are given in Section 2 above.
5. Use of the one or more nanofibres
[00103] The invention relates to a method of promoting skin care and repair in a subject, the method comprising applying to a subject in need thereof (a) one or more nanofibres formed from a
polymer and (b) an effective amount of one or more active agent, such as an active agent being, or obtained, from a plant extract, or one or more active agents comprising one or more phenolic moieties, that is associated with or chemically linked to the one or more nanofibres, the active agent selected from the group comprising
■ an active agent that assists or enhances wound healing,
■ an antibacterial active agent,
■ an anti-inflammatory active agent,
■ an active agent that assists or enhances cell proliferation,
■ an antioxidant active agent,
■ an active agent that lightens skin,
■ an active agent that preserves or improves moisture retention, and
■ an active agent that preserves or improves skin elasticity, and
■ any combination of two or more thereof,
and wherein the one or more nanofibres dissolves at a rate of at least O. mg/min when solubilised in water at 20° C.
[00104] The invention also relates to a method of promoting skin care or repair in a subject, the method comprising applying to a subject in need thereof (a) one or more nanofibres formed from a polymer and (b) an effective amount of one or more active agent being, or obtained, from a plant extract that is covalently linked to the one or more nanofibres.
[00105] In some embodiments nanofibres are formed into a matrix of fibres.
[00106] In some embodiments the nanofibre, nanofibres or matrix of nanofibres are formed onto a support.
[00107] In some embodiments the support is formed from plastic, cloth, For example, the backing may be formed from polypropylene sheets. It should be appreciated that any suitable inert backing support could be used. Suitability can be dependent on the layers compatibility with being applied to skin (i.e. non-toxic) and its ability to carry the nanofibres without any significant loss of the nanofibres from the support.
[00108] In some embodiments the nanofibre, nanofibres or matrix of nanofibres is applied to a support and used as a band aid, bandage, dressing, operating gown, facial mask, facial strip, cosmetic patch or the like.
[00109] Another aspect of the invention relates to a one or more nanofibres comprising a polymer, and an active agent obtained from a plant extract, for
■ assisting or enhancing wound healing,
■ providing an antibacterial effect,
■ providing an anti-inflammatory effect,
■ assisting or enhancing cell proliferation,
■ providing an antioxidant active agent,
■ providing a skin lightening effect,
■ preserving or improving moisture retention, or
■ preserves or improves skin elasticity.
EXAMPLES
EXAMPLE 1: WOUND HEALING MATRIX
[00110] This example describes the use of a skin patch for assisting or enhancing wound healing.
[00111] As a first step the active agent is obtained. The active agent is a plant extract and is obtained from one or more of grapes, boysenberries, blackcurrants and kiwifruit using water extraction. The active agent includes bioactives that have a phenolic moiety.
[00112] The next step is to obtain the polymer user for the formation of the nanofibres. In this particular example the polymer is collagen obtained from Hoki, a cold water fish, using the process as described in New Zealand patent 501386, published as PCT international patent application WO 2001/038396A1.
[00113] The active agent and the collagen polymer are then mixed in a solvent phase comprising water and acetic acid. It should be appreciated that any suitable solvent can be used providing it does not leave a toxic residue, solubilises both the polymer and the active agent, does not inactive the active agent, and evaporates off during electro spinning.
[00114] Once prepared the polymer-active agent solution is electrospun. The electrospinning produces a matrix of nanofibres.
[00115] The matrix of nanofibres is then applied directly to a support, such as a woven support or a plastic patch. The support or plastic patch is then applied to a wound, with the facing of the patch containing the nanofibres being applied to the skin.
[00116] The nanofibres solubilise quickly delivering the active agent and the collagen to the wound.
EXAMPLE 2: SKIN REPAIR
[00117] This example describes the use of a skin patch for assisting or enhancing skin repair.
[00118] As a first step the active agent actinidin is provided from kiwifruit (Actinidia deliciosa) using water extraction.
[00119] The next step is to provide the polymer user for the formation of the nanofibres. In this particular example the polymer is marine collagen obtained from Hoki, a cold water fish, using the process as described in New Zealand patent 501386, published as PCT international patent application WO 2001/038396A1.
[00120] The active agent and the collagen polymer are then mixed in a solvent phase comprising water and acetic acid.
[00121] Once prepared the polymer-active agent solution is electrospun. The electrospinning produces a matrix of nanofibres.
[00122] The matrix of nanofibres is then applied directly to a support, such as a woven support or a plastic patch. The support or plastic patch is then applied to the skin, with the facing of the patch containing the nanofibres being applied to the skin.
[00123] The nanofibres solubilise quickly delivering the actinidin and the collagen to the skin. EXAMPLE 3: BIOACTIVE DRESSING/PATCH
[00124] This example analyses the efficacy of a dressing or patch that contained bioactive extracts that have skin repair and skin care functions. The bioactives were contained within a layer or layers of nanofibres that dissolved or released the actives at different rates.
[00125] The active agent in this example comes from plant sources that include grapes, berries, kiwifruit and other plant material. The plant extract contained concentrated levels of bioactives including carotenoids, phenolics and enzymes.
[00126] Two different matrices of nanofibres were used.
(1) A matrix formed from PVOH or PA-6 nanofibres impregnated with Vinanza Skin Performance Plus Extract as obtained from New Zealand Extracts Limited.
(2) A matrix formed from PVOH nanofibres and impregnated with Vinanza Skin Repair Plus as obtained from New Zealand Extracts Limited.
[00127] The nanofibres were spun to form layers of nanofibres on polypropylene sheets.
[00128] The sheets were then incubated in Hanks solution and the release of the bioactives was measured over time.
[00129] The different nanofibre materials released the bioactives at different rates. For example, phenolic bioactives were released from PVOH nanofibres within two minutes whereas with PA-6 nanofibres the bioactives were released from the fibres over several days.
[00130] Shown in Figure 1 is the release of bioactives from a nanofibre matrix impregnated with Vinanza Skin Performance Plus Extract. As can be seen in Figure 1, the nanofibre formed from PVOH released the phenolic bioactive effectively instandy, while the nanofibre formed from PA-6 released the phenolic over a much longer period of time.
[00131] The observed kinetics of release of the individual phenolic active agents 3-pCoQA (♦), catechin (■), procyanidinB2 (A), and epi-catechin (X) from the PVOH nanofibre is shown in Figure 2. The observed kinetics of release of the phenolic active agents 3-CQA (♦), 3-pCoQA (■), catechin (A), procyanidinB2 (X), and epi-catechin (*) from PA-6 matrix is shown in Figure 3. As can be seen, the release of these phenolic compounds from each matrix varies significandy.
[00132] Shown in Figure 4 is the effect of Vinanza Grape Seed Extract on (1) trans epidermal water loss and (2) skin elasticity, and the effect of Vinanza Kiwifruit Extract on (1) skin redness, and (2) skin melanin content.
[00133] As can be seen in Figure 4, Vinanza Grape Seed Extract improved skin cell integrity by decreasing trans epidermal water loss (TEWL) by at least 6%, and improved skin elasticity by 5%. Also demonstrated is that Vinanza Kiwifruit Extract decreased skin redness by 7% and lightened skin by 3.5%.
[00134] Shown in Figure 5 is Vinanza Skin Repair Plus applied as a PVOH nanofibres on the repair of skin damaged by the application of sodium lauryl sulphate (SLS). Here, the percent change was the difference between the marker of skin damage on the damaged skin, versus the same marker on undamaged skin, where a lower % change represents less damage.
[00135] As can clearly be seen in Figure 5, the application of a patch comprising a nanofibre matrix of the invention, in this case comprising Vinanza Skin Repair Plus extracts, was very effective in minimising skin damage and speeding skin repair.
EXAMPLE 4: WOUND HEALING MATRIX
[00136] This example describes the preparation and use of a skin patch for assisting or enhancing wound healing.
[00137] Blackcurrant and Red grape extracts comprising active agents including one or more antimicrobial agents, and collagen (as described above in Example 1) are obtained. The extracts comprising one or more antimicrobial active agents and the collagen polymer are then mixed in a
solvent phase. Separately, grape seed extract comprising one or more skin repair active agents and PA-6 polymer are mixed in a solvent phase.
[00138] Once prepared, the polymer-active agent solutions are electro spun. The electro spinning produces a matrix of nanofibres, wherein the matrix comprises both collagen (water- soluble) and PA-6 (effectively water-insoluble) polymers.
[00139] The matrix of nanofibres is applied directly to a support, such as a woven bandage or patch, suitable for application. In this case, the electro spinning is performed direcdy onto the bandage or wound dressing.
[00140] The dressing is then packaged for storage prior to application to a wound. On application, a subset of the nanofibres, typically some or all of the water soluble nanofibres solubilise quickly to essentially immediately deliver one or more of the antimicrobial active agents and the collagen to the wound. Release of the active agents effective for skin repair from the PA-6 polymer then occurs over a longer time period, typically over several days.
EXAMPLE 5: SKIN REPAIR
[00141] This example describes the use of a skin patch for assisting or enhancing skin repair.
[00142] Grape seed extracts comprising active agents including one or more skin repair active agents, and collagen (as described above in Example 1) are obtained. The grape seed extract and the collagen polymer are then mixed in a solvent phase. Separately, the grape seed extract and PA-6 polymer are mixed in a solvent phase.
[00143] Once prepared, the polymer-active agent solutions are electro spun. The electro spinning produces a matrix of nanofibres, wherein the matrix comprises both water-soluble and water-insoluble polymers.
[00144] The matrix of nanofibres is applied directly to a support, such as a woven bandage or patch. In this example, the electro spinning is performed direcdy onto a patch.
[00145] The patch is then packaged for storage prior to application to damaged or sunburned skin. On application to skin, a subset of the nanofibres, typically some or all of the collagen nanofibres solubilise quickly to essentially immediately deliver one or more of the active agents present in the grape seed extract and the collagen to the skin for immediate effect. Release of additional active agents from the PA-6 polymer then occurs over a longer time period, from one hour to several days, thereby maintaining enhanced skin repair for the duration of application of the patch.
[00146] Where in the foregoing description reference has been made to elements or integers having known equivalents, then such equivalents are included as if they were individually set forth.
[00147] Although the invention has been described by way of example and with reference to particular embodiments, it is to be understood that modifications and/ or improvements may be made without departing from the scope or spirit of the invention.
[00148] Where in the foregoing description reference has been made to elements or integers having known equivalents, then such equivalents are included as if they were individually set forth.
[00149] Although the invention has been described by way of example and with reference to particular embodiments, it is to be understood that modifications and/ or improvements may be made without departing from the scope or spirit of the invention.
INDUSTRIAL APPLICABILITY
The present invention provides active agent delivery systems, and methods of using these systems, for example in the treatment and prevention of skin diseases or conditions, and as such is expected to provide both social and economic benefits