Formulation for topical wound treatment

Abstract

The present invention relates to new formulations, particularly useful in the topical treatment of skin and oromucosal wounds. Formulations can be either a solution or a gel form consisting of a Polyhexamethylene biguanide as an antimicrobial agent, purified water and a tri-blockcopolymer, particularly a triblockcopolymer of polyethylene oxide and polypropylene oxide, and more particularly a poloxamer.

Claims

1. A formulation, consisting of polyhexamethylene biguanide, purified water, humectant, gelling agent and 1.0% by weight of at least one poloxamer for use in the topical and/or oromucosal treatment of wounds; wherein the humectant is selected from beta-sitosterol, inositol, glycerin, urea, hydroxyethyl urea, glyceryl stearate, panthenol, propylene glycol, polyethylene glycol and xylitol; and wherein the gelling agent is selected from methyl cellulose, hydroxypropyl methyl cellulose, carboxymethylcellulose, hydroxyethyl methylcellulose, hydroxyethyl cellulose, sodium carboxymethylcellulose, acrylic acid polymers, and non-ionic poly(ethylene oxide) polymers.

2. The formulation for the use in the topical and/or oromucosal treatment of wounds according to claim 1, wherein the poloxamer is selected from poloxamer 124, poloxamer 188, poloxamer 237, poloxamer 338 and poloxamer 407.

3. The formulation for the use in the topical and/or oromucosal treatment of wounds according to claim 1, wherein the poloxamer is poloxamer 188.

4. The formulation for the use in the topical and/or oromucosal treatment of wounds according to claim 1, consisting of 0.1% polyhexamethylene biguanide (PHMB) by weight, 1.0% poloxamer by weight, 8.6% humectant by weight, 1.8% hydroxyethyl cellulose by weight, and 88.5% purified water by weight.

5. The formulation for the use in the topical and/or oromucosal treatment of wounds according to claim 1, wherein the humectant is glycerin or hydroxyethyl urea.

6. The formulation for the use in the topical and/or oromucosal treatment of wound according to claim 1, wherein the gelling agent is selected from methyl cellulose, hydroxypropyl methyl cellulose, carboxymethylcellulose, hydroxyethyl methylcellulose, hydroxyethyl cellulose, and sodium carboxymethylcellulose.

Description

FORMULATIONS ACCORDING TO THE PRESENT INVENTION

(1) Formulations According to the Present Invention Comprising PHMB and Tri-Blockcopolymers

(2) The formulation is preferably provided in two forms, which are gel and solution:

(3) The formulation used for topical and oromucosal treatment of wounds according to the present invention comprise polyhexamethylene biguanide (PHMB) as active agent, purified water as solvent and at least one tri-blockcopolymer according to the present invention.

(4) The tri-blockcopolymers according to the present invention are amphiphilic. They further are non-toxic and thus can be added to formulations for wound treatment. The tri-blockcopolymers further are non-ionic, such that they do not interfere with the active agent PHMB, as compared to anionic surfactants that reduce the bactericidal activity of PHMB due to reacting with it. Furthermore the tri-blockcopolymers are non-ionic, independent from the surrounding pH. Thus the tri-blockcopolymers can be used for the treatment of open wounds that often have a basic environment or in combination with acid components that are used to generate an acidic environment on the wound, such that wound healing can start.

(5) Particularly preferred tri-blockcopolymers are tri-blockcopolymers of polyethylene oxide and polypropylene oxide. Said tri-blockcopolymers of polyethylene oxide and polypropylene oxide are amphiphilic, non-toxic and non-ionic.

(6) One kind of tri-blockcopolymers of polyethylene oxide and polypropylene oxide are poloxamers, which are particularly preferred in the present invention. Particularly preferred poloxamers used in the present invention can be selected from poloxamer 124, poloxamer 188, poloxamer 237, poloxamer 338 and poloxamer 407, whose properties are summarized in Table 2 above. Among these poloxamers, poloxamer 188 is particularly preferred. However, several different tri-blockcopolymers of polyethylene oxide and polypropylene oxide or some other co-surfactants (i.e. Panthenyl hydroxypropyl steardimonium chloride, Cas No: 132467-76-6) described above may be used in combination.

(7) In a preferred aspect, the formulation is a solution comprising 0.05-10.0% Polyhexamethylene biguanide (PHMB) by weight, 0.1-40% tri-blockcopolymer by weight and completed to 100% by adding purified water in order to get the final wound treatment solution.

(8) In yet another preferred aspect, the formulation is a gel comprising 0.05-.,0% Polyhexamethylene biguanide (PHMB) by weight, 0.1-40% tri-blockcopolymer by weight, 1.0-10.0% Humectant by weight, 0.5-5.0% gelling agent by weight and completed to 100% by adding purified water in order to get the final wound treatment solution.

(9) In another preferred aspect, the humectant is selected from glycerine and hydroxyethyl urea.

(10) In another preferred aspect, the formulation comprises at least one co-surfactant. Such co-surfactants may improve the solubility of any herbal ingredients that may be added or to further reduce the cytotoxicity of the formulation.

(11) The preferred concentration of co-surfactants in any formulation is 0-2% by weight, more preferably 0.01-2% by weight, even more preferably 0.05-1% by weight and most preferably 0.1% by weight.

(12) The co-surfactants according to the present invention can be chosen freely, as long as they are suitable for contact with the skin and wounds and, do not cause any increase in cytotoxicity or induce any damage to the wound.

(13) The co-surfactants according to the present invention are preferably selected from glycine, sulfosuccinate and amide derivatives of an unbranched fatty acid, alkyl betaines, alkyl sultaines, alkyl amine oxides, alkyl imidazolium derivatives, unbranched fatty acid derivatives of amine oxide and unbranched fatty acid derivatives of imidazolium. Among these, the co-surfactants according to the present invention are more preferably selected from alkyl betaines, alkyl sultaines, alkyl amine oxides and alkyl imidazolium derivatives. On the other hand, betaines type surfactants can be selected from fatty alcohol or fatty amine derivatives. The alkyl group in said compounds can be branched or unbranched having 8-18 carbon atoms, preferably 10-16 carbon atoms and most preferably 10-14 carbon atoms. Moreover the alkyl group can be either saturated or unsaturated carbon chain.

(14) Of said co-surfactants, alkyl betaines, alkyl sultaines, alkyl amine oxides, alkyl imidazolium derivatives and panthenol derivatives are preferred as the co-surfactants for the present formulations. The panthenol derivative surfactants are particularly preferred as the co-surfactants for the present formulations.

(15) The alkyl betaines are preferably selected from Lauryldimethylbetaine, Cocobetaine, Myristyl betaine, Decyl betaine, Lauryl sultaine, Oleyldimethylbetaine, Dodecylbetaine, Caprylyl betaine, and Behenyl betaine. The alkyl amine oxides are preferably selected from Lauramine oxide, Lauramidopropylamine oxide, Cocamine oxide, Cocamidopropylamine

(16) Oxide, Myristamine oxide, and Soyamidopropylamine Oxide. The imidazoline derivatives are preferably selected from Disodium cocoamphodiacetate, Disodium lauroamphodiacetate, Sodium lauroamphoacetate, Disodium cocoamphodipropionate and Sodium cocoamphoacetate. The panthenol derivative is preferably selected from Panthenyl hydroxypropyl steardimonium chloride.

(17) In a preferred aspect, the formulation may further comprise a substance/substances obtained from herbs, as described above. Such substances obtained from herbs may have the beneficial properties described above in the according section.

(18) Thus, in a preferred aspect, the formulation in form of a solution or a gel further comprises a substance/substances obtained from herbs. The substance/substances obtained from said herbs is/are preferably Comfrey (Symphytum officinale L) herb extract and/or Myrrh (Commiphora molmol) tincture. It is noted that the formulations according to the present invention also has beneficial effects compared to those commonly used when no such substance/substances obtained from said herbs is/are used. However, the addition of substance/substances obtained from said herbs may further improve the effect of the formulations according to the present invention.

(19) The substance/substances obtained from herbs is/are preferably Comfrey herb extract and/or Myrrh tincture. The Comfrey herb extract is preferably obtained from the leaves of comfrey herb.

(20) However, the formulations according to the present invention do not require the presence of substances obtained from herbs. Also a formulation in form of a solution or gel as described above, not containing any substances obtained from herbs can have the desired effect for the treatment of wounds. In this respect it is noted again that the tri-blockcopolymers, in particular poloxamers according to the present invention, are amphiphilic, non-toxic and non-ionic surfactants and thus can be used in combination with PHMB for the treatment of wounds with the desired beneficial effects.

(21) The use of additional substances obtained from herbs may thus simply further increase the beneficial effect already obtained by using PHMB with tri-blockcopolymers according to the present invention.

(22) Accordingly, in one aspect, the formulation in form of a solution or a gel thus does not contain any substance/substances obtained from herbs. As described above, substance/substances obtained from herbs may have beneficial effects for the treatment of wounds. However it has been recognized that also PHMB with poloxamer alone (i.e. without any substance/substances obtained from herbs) can have the desired effect on wounds.

(23) Thus, a formulation comprising PHMB and poloxamer in addition with other excipients such as purified water, co-surfactants, humectants and gelling agents may be used, without adding any substance/substances obtained from herbs.

(24) Thus, in a preferred aspect, the formulation in form of a solution only consists of polyhexamethylene biguanide (PHMB) as active agent, purified water as solvent, at least one tri-blockcopolymer and at least one co-surfactant chosen among those described above.

(25) In yet another preferred aspect, the formulation in form of a solution only consists of polyhexamethylene biguanide (PHMB) as active agent, purified water as solvent and at least one tri-blockcopolymer.

(26) The formulation may thus consist of 0.05-10.0% PHMB by weight, 0.1-40% tri-blockcopolymer by weight and completed to 100% by adding purified water in order to get the final wound treatment solution. The formulation in form of a solution may thus consist of 0.1% PHMB by weight, 1.0% tri-blockcopolymer (such as poloxamer 188) by weight and 98.9% purified water by weight.

(27) Thus, in a preferred aspect, the formulation in form of a gel only consists of polyhexamethylene biguanide (PHMB) as active agent, purified water as solvent, humectant, gelling agent, at least one tri-blockcopolymer and at least one co-surfactant chosen among those described above.

(28) The formulation may thus consist of 0.05-10.0% PHMB by weight, 0.1-40% tri-blockcopolymer by weight, 0.01-2% co-surfactant by weight and completed to 100% by adding purified water in order to get the final wound treatment solution. The formulation in form of a solution may thus consist of 0.1% PHMB by weight, 1.0% tri-blockcopolymer by weight, 0.1% co-surfactant by weight and 98.8% purified water by weight.

(29) In yet another preferred aspect, the formulation in form of a gel only consists of polyhexamethylene biguanide (PHMB) as active agent, purified water as solvent, humectant, gelling agent and at least one tri-blockcopolymer.

(30) The gelling agents are preferably selected from polymers usually used in gel formulations to decrease the mobility and increase the viscosity of a product. Gelling agent are preferably a cellulose derivative or another commercially available gelling agent. Preferred gelling agents are cellulose derivatives (e.g. methyl cellulose, hydroxypropyl methyl cellulose, carboxymethylcellulose, hydroxyethyl cellulose) and/or modified acrylic acid polymers (e.g. carbopol), either alone or in combination. Particularly preferred gelling agents are Methylcellulose, Hydroxypropyl Methylcellulose, Hydroxyethyl Methylcellulose, Hydroxyethyl cellulose, Carboxymethylcellulose and Sodium Carboxymethylcellulose. Other preferred gelling agents are non-ionic poly (ethylene oxide) polymers. Another preferred gelling agent is carboxypolymethylene, also known as Carbopol, which is a modified acrylic acid polymer, preferably selected from Carbopol 940, Carbopol 934 [USAN:NF], Carbopol 907, Carbopol 941, Carbopol Ultrez 10 and Carbopol 5984.

(31) Although preferred gelling agents are to be in non-ionic form, anionic molecules (i.e. carboxymethylcellulose and carbopol) can also be used in the formulation in the presence of a cationic excipient, which may be a co-surfactant (e.g. Panthenyl hydroxypropyl steardimonium chloride.

(32) The Humectants are preferably chosen from beta-sitosterol, inositol, glycerin, urea, hydroxyethyl urea, glyceryl stearate, panthenol, propylene glycol, polyethylene glycol and its derivatives (i.e. methyl gluceth 10 and methyl gluceth 20), and Xylitol. The humectants are even more preferably selected from glycerin and hydroxyethyl urea.

(33) The formulation consists in a preferred aspect of 0.05-2.0% PHMB by weight, 0.1-40% tri-blockcopolymer by weight, 1.0-10.0% Humectant by weight, 0.5-5.0% gelling agent by weight and completed to 100% by adding purified water in order to get the final wound treatment solution. The formulation in form of a gel may thus consist of 0.1% PHMB by weight, 1.0% tri-blockcopolymer (such as poloxamer 188) by weight, 8.6% glycerin by weight, 1.8% by weight hydroxyethyl cellulose by weight and 88.5% purified water by weight.

(34) The formulation consists in another preferred aspect of 0.05-2.0% PHMB by weight, 0.1-40 tri-blockcopolymer by weight, 0.01-2% co-surfactant by weight, 1.0-10.0% Humectant by weight, 0.5-5.0% gelling agent by weight and completed to 100% by adding purified water in order to get the final wound treatment solution. The formulation in form of a gel may thus consist of 0.1% PHMB by weight, 1.0% tri-blockcopolymer (such as poloxamer 188) by weight, 0.1% co-surfactant by weight, 8.6% glycerin by weight, 1.8% by weight hydroxyethyl cellulose by weight and 88.4% purified water by weight.

(35) Another aspect of the present invention is a method for producing a topical and oromucosal treatment formulation for the treatment of wounds, the method comprising the process steps of adding water to the mixer, adding and mixing polyhexamethylene biguanide (PHMB) and purified water (solution A) and adding and mixing said solution A to tri-blockcopolymer to get the end product.

(36) Another aspect of the present invention is a method for producing a topical and oromucosal treatment formulation for the treatment of wounds, the method comprising the process steps of adding gelling agent to distilled water and mixing thereof by heating followed by adding PHMB (solution B), cooling said solution B and adding and mixing said cooled solution B to tri-blockcopolymer to get the end product.

(37) The formulation according to the present invention may only contain the above-described PHMB and tri-blockcopolymers, purified water, humectants (e.g. glycerin, inositol, panthenol, xylitol . . . etc.), gelling agents (e.g. carbopol, carboxymethylcellulose, hydroxyethyl cellulose, methyl cellulose . . . etc.), i.e. without any additional compounds such as substance/substances obtained from herbs.

(38) Exemplary Formulations

(39) Formulations A, B and C Containing PHMB and Tri-blockcopolymers

(40) In the following, two exemplary formulations according to the present invention, one in the form of a solution and one in the form of a gel are described. These formulations are also compared later to commonly known formulations in the experimental section.

(41) Exemplary Formulation A of a solution according to the present invention:

(42) TABLE-US-00005 Component Content (g) Function PHMB* 0.1 g (= 0.1 w.-%) Active substance Poloxamer 188 1.0 g (= 1.0 w.-%) Auxiliary substance Distilled water Ad 100 Solvent *20% PHMB (0.5 g) are used; 0.5 20/100 = 0.1 g

(43) Exemplary Formulation B of a gel according to the present invention:

(44) TABLE-US-00006 Component Content (g) Function PHMB* 0.1 g (= 0.1 w.-%) Active substance Poloxamer 188 1.0 g (= 1.0 w.-%) Auxiliary substance Glycerin 8.6 g (= 8.6 w.-%) Auxiliary substance Hydroxyethyl cellulose 1.8 g (= 1.8 w.-%) Auxiliary substance Distilled water Ad 100 Solvent *20% PHMB (0.5 g) are used; 0.5 20/100 = 0.1 g

(45) Exemplary Formulation C of a gel according to the present invention:

(46) TABLE-US-00007 Component Content (g) Function PHMB* 0.1 g (= 0.1 w.-%) Active substance Poloxamer 188 1.0 g (= 1.0 w.-%) Auxiliary substance Hydroxyethyl Urea 8.6 g (= 8.6 w.-%) Auxiliary substance Hydroxyethyl cellulose 1.8 g (= 1.8 w.-%) Auxiliary substance Distilled water Ad 100 Solvent *20% PHMB (0.5 g) are used; 0.5 20/100 = 0.1 g

(47) Formulations Comprising PHMB, Tri-blockcopolymers and Co-surfactants

(48) Formulations according to the present invention may also include co-surfactants as an additional excipient either to improve the solubility of any added herbal contents or to incorporate additional synergistic effects to the formulation. Following examples, one in the form of a solution and one in the form of a gel, are describing the basic formulation with co-surfactants.

EXAMPLE 1

(49) By mixing the following ingredients listed in the usual way a wound solution is made for cleaning and/or treating wounds.

(50) TABLE-US-00008 Component Amount (wt. %) PHMB 0.1 Poloxamer 1.0 Cocobetaine 0.1 Purified Water 98.8

EXAMPLE 2

(51) By mixing the following ingredients listed in the usual way a wound solution is made for cleaning and/or treating wounds.

(52) TABLE-US-00009 Component Amount (wt. %) PHMB 0.1 Poloxamer 1.0 Lauramine oxide 0.1 Purified Water 98.8

EXAMPLE 3

(53) By mixing the following ingredients listed in the usual way a wound solution is made for cleaning and/or treating wounds.

(54) TABLE-US-00010 Component Amount (wt. %) PHMB 0.1 Poloxamer 1.0 Sodium cocoamphoacetate 0.1 Purified Water 98.8

EXAMPLE 4

(55) By mixing the following ingredients listed in the usual way a wound solution is made for cleaning and/or treating wounds.

(56) TABLE-US-00011 Component Amount (wt. %) PHMB 0.1 Poloxamer 1.0 Cocobetaine 0.1 PHSC* 0.1 Purified Water 98.7 *Panthenyl hydroxypropyl steardimonium chloride

EXAMPLE 5a

(57) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(58) TABLE-US-00012 Component Amount (wt. %) PHMB 0.1 Poloxamer 1.0 Cocobetaine 0.1 Glycerin 8.6 Hydroxyethyl cellulose 1.8 Purified Water 88.4

EXAMPLE 5b

(59) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(60) TABLE-US-00013 Component Amount (wt. %) PHMB 0.1 Poloxamer 1.0 Cocobetaine 0.1 Hydroxyethyl Urea 8.6 Hydroxyethyl cellulose 1.8 Purified Water 88.4

EXAMPLE 6a

(61) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(62) TABLE-US-00014 Component Amount (wt. %) PHMB 0.1 Poloxamer 1.0 Lauramine oxide 0.1 Glycerin 8.6 Hydroxyethyl cellulose 1.8 Purified Water 88.4

EXAMPLE 6b

(63) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(64) TABLE-US-00015 Component Amount (wt. %) PHMB 0.1 Poloxamer 1.0 Lauramine oxide 0.1 Hydroxyethyl Urea 8.6 Hydroxyethyl cellulose 1.8 Purified Water 88.4

EXAMPLE 7a

(65) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(66) TABLE-US-00016 Component Amount (wt. %) PHMB 0.1 Poloxamer 1.0 Sodium cocoamphoacetate 0.1 Glycerin 8.6 Hydroxyethyl cellulose 1.8 Purified Water 88.4

EXAMPLE 7b

(67) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(68) TABLE-US-00017 Amount Component (wt. %) PHMB 0.1 Poloxamer 1.0 Sodium cocoamphoacetate 0.1 Hydroxyethyl Urea 8.6 Hydroxyethyl cellulose 1.8 Purified Water 88.4

EXAMPLE 8a

(69) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(70) TABLE-US-00018 Amount Component (wt. %) PHMB 0.1 Poloxamer 1.0 Cocobetaine 0.1 PHSC* 0.1 Glycerin 8.6 Hydroxyethyl cellulose 1.8 Purified Water 88.3 *Panthenyl hydroxypropyl steardimonium chloride

EXAMPLE 8b

(71) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(72) TABLE-US-00019 Amount Component (wt. %) PHMB 0.1 Poloxamer 1.0 Cocobetaine 0.1 PHSC* 0.1 Hydroxyethyl Urea 8.6 Hydroxyethyl cellulose 1.8 Purified Water 88.3 *Panthenyl hydroxypropyl steardimonium chloride

(73) Formulations Comprising PHMB, Tri-blockcopolymers and Further Substances Obtained from Herbs

(74) Further to the above-described compounds, the formulations may contain substance/substances obtained from herbs, as described above. Such substances obtained from herbs may have the beneficial properties described above in the according section.

(75) However, the formulations according to the present invention do not require the presence of substances obtained from herbs. Also a formulation in form of a solution of gel as described above, not containing any substances obtained from herbs can have the desired effect for the treatment of wounds. In this respect it is noted again that the tri-blockcopolymers, in particular poloxamers according to the present invention, are amphiphilic, non-toxic and non-ionic surfactants and thus can be used in combination with PHMB for the treatment of wounds with the desired beneficial effects.

(76) The use of additional substances obtained from herbs may thus simply further increase the beneficial effect already obtained by using PHMB with tri-blockcopolymers according to the present invention.

(77) Formulations in Form of a Solution Comprising PHMB, Tri-Blockcopolymers and Further Substances Obtained from Herbs

(78) The present invention thus further comprises a solution formulation containing one or more herbal substance, PHMB, an emulsifying agent (a tri-blockcopolymer), and purified water to use for the topical treatment of skin and oromucosal wounds.

(79) The solution formulation is intended to be used for a mixture preparation composed of PHMB, purified water, an emulsifying agent and/or Comfrey (Symphytum officinale L.) extract and/or Myrrh (Commiphora molmol) tincture, and optionally a co-surfactant or a mixture of co-surfactants.

(80) The preferred concentration of Comfrey (Symphytum officinale L.) extract in any solution formulation is 1.0-15.0% (w/w), more preferably 3.0-12.0% (w/w), and most preferably 10.0% (w/w).

(81) The preferred concentration of Myrrh (Commiphora molmol) tincture in any solution formulation is 1.0-20.0% (w/w), more preferably 3.0-12.0% (w/w), and most preferably 5.0% (w/w).

(82) The preferred concentration of Polyhexamethylene biguanide (PHMB) in any solution formulation is 0.05-2.0% (w/w), more preferably 0.08-0.3% (w/w), and most preferably 0.1% (w/w).

(83) The preferred concentration of emulsifying agent in any solution formulation is 0.1-40.0% (w/w), more preferably 0.5-20% (w/w), and most preferably 1.0% (w/w). The preferred concentration of co-surfactants in any solution formulation is 0-2% (w/w), more preferably 0.01-2% (w/w), even more preferably 0.05-1% (w/w), and most preferably 0.1% (w/w). The mixture prepared by mixing the ingredients above listed is completed to 100% by adding distilled water in order to get final wound treatment solution.

(84) TABLE-US-00020 TABLE 3 Usable and preferred quantities of ingredients in a solution formulation. Usable Preferred quantity quantity by by weight weight Substance Function (%) (%) Herbal content 1 Anti-inflammatory and 1.0-15.0 10.0 Comfrey analgesic effects, it induces (Symphytum granulation and is used for officinale L) wound treatment as it Extract supports tissue regeneration Herbal content 2 Antibacterial, antifungal 1.0-20.0 5.0 Myrrh and local anaesthetic effects (Commiphora molmol) tincture Poloxamer Emulsifying agent; 0.1-40.0 1.0 dispersing agent; wetting agent; solubilizing agent; lubricant, detergents Polyhexamethylene Protective agent; algaecides, 0.05-2.0 0.1 biguanide (PHMB) bactericides/bacteriostatics, fungicides/fungistatics. microbicides/microbiostatics. disinfectants, regulators, and sanitizers. Purified Water Diluting agent/ 23.0-98.0 73.5 supplementary agent Co-surfactant improve solubility of 0.01-2.0 0.1 herbal ingredients, reduce the cytotoxicity of the formulation

(85) In order to fulfill the aforementioned objectives the production steps of the solution formulation is as follows: adding and mixing herbal substance/substances, Poloxamer and optionally a co-surfactant or a mixture of co-surfactants for providing Solution X, separately adding and mixing polyhexamethylene biguanide (PHMB) and distilled water for providing

(86) Solution Y and finally; adding and mixing Solution X to Solution Y to get the end product. After that the final solution obtained after the process, is objected to filtration.

(87) Production of various solutions formulations according to invention further including substances obtained from herbs is described in detail as examples below:

EXAMPLE 1

(88) By mixing the following ingredients listed in the usual way a wound solution is made for cleaning and/or treating wounds.

(89) TABLE-US-00021 Comfrey 10.0 (Symphytum officinale L) Extract Myrrh 5.0 (Commiphora molmol) Tincture Poloxamer 1.0 Polyhexamethylene 0.1 biguanide (PHMB) Purified Water 83.9

EXAMPLE 2

(90) By mixing the following ingredients listed in the usual way a wound solution is made for cleaning/ treating wounds.

(91) TABLE-US-00022 Comfrey 10.0 (Symphytum officinale L) Extract Poloxamer 1.0 Polyhexamethylene 0.1 biguanide (PHMB) Purified Water 88.9

EXAMPLE 3

(92) By mixing the following ingredients listed in the usual way a wound solution is made for cleaning/ treating wounds.

(93) TABLE-US-00023 Myrrh 5.0 (Commiphora molmol) Tincture Poloxamer 1.0 Polyhexamethylene 0.1 biguanide (PHMB) Purified Water 93.9

(94) Formulations in Form of a Gel Comprising PHMB, Tri-blockcopolymers and Further Substances Obtained from Herbs:

(95) The present invention further comprises a gel formulation containing one or more herbal substance, PHMB, an emulsifying agent (a tri-block copolymer), a gelling agent, a humectant and purified water to use for the topical treatment of skin and oromucosal wounds.

(96) The gel formulation is intended to be used for a mixture preparation composed of PHMB, purified water, an emulsifying agent, a gelling agent, a humectant and/or Comfrey (Symphytum officinale L.) extract and/or Myrrh (Commiphora molmol) tincture, and optionally a co-surfactant or a mixture of co-surfactants.

(97) The preferred concentration of Comfrey (Symphytum officinale L.) extract in any gel formulation is 1.0-15.0% (w/w), more preferably 3.0-12.0% (w/w), and most preferably 10% (w/w).

(98) The preferred concentration of Myrrh (Commiphora molmol) tincture in any gel formulation is 1.0-20.0% (w/w), more preferably 3.0-12.0% (w/w), and most preferably 5.0% (w/w).

(99) The preferred concentration of Polyhexamethylene biguanide (PHMB) in any gel formulation is 0.05-2.0% (w/w), more preferably 0.08-0.3% (w/w), and most preferably 0.1% (w/w).

(100) The preferred concentration of emulsifying agent in any gel formulation is 0.1-40.0% (w/w), more preferably 0.5-20% (w/w), and most preferably 1.0% (w/w).

(101) The preferred concentration of gelling agent in any gel formulation is 0.5-5.0% (w/w), more preferably 1.0-3.0% (w/w), and most preferably 1.8% (w/w).

(102) The preferred concentration of a humectant in any gel formulation is 1.0-10.0% (w/w), more preferably 5.0-9 .0% (w/w), and most preferably 8.6% (w/w). The preferred concentration of co-surfactants in any gel formulation is 0-2% (w/w), more preferably 0.01-2% (w/w), even more preferably 0.05-1% (w/w), and most preferably 0.1% (w/w). The mixture prepared by mixing the ingredients above listed completed to 100% by adding distilled water in order to get final wound treatment gel.

(103) TABLE-US-00024 TABLE 4 Gel content and preferred/usable quantities by weight Usable Preferred quantity quantity by by weight weight Substance Function (%) (%) Herbal content 1 Anti-inflammatory 1.0-15.0 10.0 Comfrey (Symphytum and analgesic effects, officinale L) Extract it induces granulation and is used for wound treatment as it supports tissue regeneration Herbal content 2 antibacterial, 1.0-20.0 5.0 Myrrh (Commiphora antifungal molmol) tincture and local anaesthetic effects Poloxamer Emulsifying agent; 0.1-40.0 1.0 dispersing agent; wetting agent; solubilizing agent; lubricant, detergents Polyhexamethylene Protective agent; 0.05-2.0 0.1 biguanide (PHMB) algaecides, bactericides/ bacteriostatics, fungicides/ fungistatics, microbicides/ microbiostatics, disinfectants, regulators, and sanitizers. Glycerin or Humectant 1.0-10.0 8.6 Hydroxyethyl urea Hydroxyethylcellulose Gelling agent 0.5-5.0 1.8 Purified Water Diluting 8.0-96.3 73.5 agent/ supplementary agent Co-surfactant improve solubility 0.01-2.0 0.1 of herbal ingredients, reduce the cytotoxicity of the formulation

(104) In order to fulfill the aforementioned objectives the production steps of the gel formulation is as follows: adding and mixing herbal substance/substances, poloxamer and optionally a co-surfactant or a mixture of co-surfactants for providing Solution X, separately adding gelling agent to purified water and mixing thereof by heating and followed by adding PHMB for providing Solution Z, cooling the Solution Z and finally; adding and mixing Solution X to Solution Z to get the end product. After that the final solution, obtained after the process, is objected to filtration.

(105) Production of various gel formulations according to invention further including substances obtained from herbs is described in detail as examples below:

EXAMPLE 4a

(106) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(107) TABLE-US-00025 Comfrey (Symphytum officinale L) Extract 10.0 Myrrh (Commiphora molmol) Tincture 5.0 Poloxamer 1.0 Polyhexamethylene biguanide (PHMB) 0.1 Glycerin 8.6 Hydroxyethyl cellulose 1.8 Purified Water 73.5

EXAMPLE 4b

(108) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(109) TABLE-US-00026 Comfrey (Symphytum officinale L) Extract 10.0 Myrrh (Commiphora molmol) Tincture 5.0 Poloxamer 1.0 Polyhexamethylene biguanide (PHMB) 0.1 Hydroxyethyl urea 8.6 Hydroxyethyl cellulose 1.8 Purified Water 73.5

EXAMPLE 5a

(110) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(111) TABLE-US-00027 Comfrey (Symphytum officinale L) Extract 10.0 Poloxamer 1.0 Polyhexamethylene biguanide (PHMB) 0.1 Glycerin 8.6 Hydroxyethylcellulose 1.8 Purified Water 78.5

EXAMPLE 5b

(112) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(113) TABLE-US-00028 Comfrey (Symphytum officinale L) Extract 10.0 Poloxamer 1.0 Polyhexamethylene biguanide (PHMB) 0.1 Hydroxyethyl urea 8.6 Hydroxyethylcellulose 1.8 Purified Water 78.5

EXAMPLE 6a

(114) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(115) TABLE-US-00029 Myrrh (Commiphora molmol) Tincture 5.0 Poloxamer 1.0 Polyhexamethylene biguanide (PHMB) 0.1 Glycerin 8.6 Hydroxyethylcellulose 1.8 Purified Water 83.5

EXAMPLE 6b

(116) By mixing the following ingredients listed in the usual way a wound gel is made for cleaning/treating wounds.

(117) TABLE-US-00030 Myrrh (Commiphora molmol) Tincture 5.0 Poloxamer 1.0 Polyhexamethylene biguanide (PHMB) 0.1 Hydroxyethyl urea 8.6 Hydroxyethylcellulose 1.8 Purified Water 83.5

(118) Application Areas

(119) The present formulations can be used for cleaning, decontamination, irrigation and moistening of infected, dried and disintegrated (chronic and acute) skin and mucosa; it is also used before application of bandages, gauzes, compresses, wound fillers and other absorbent materials.

(120) Experimental Results

(121) General Results with Formulations Containing PHMB and Poloxamer 188 According to Formulations A and B

(122) It has been shown that the solution (Exemplary Formulation A) according to the present invention has good decolonization properties for Methicillin-resistant S. aureus (MRSA). Methicillin-resistant S. aureus is a gram positive bacterial strain, which causes pneumonia, wound and hospital infections. They are resistant to several antibiotics. The solution according to the present invention has a strong bactericidal effect on MRSA and can easily be applied onto the infected area.

(123) The same results were achieved for Vancomycin-resistant Enterococci (VRE), another strain of gram positive bacteria, which cause severe hospitals infections mainly in chronic hemodialysis patients. Also here the according to the present invention has a strong bactericidal effect on VRE and can easily be applied onto the infected area.

(124) Also for Acinetobacter baumannii, gram negative pleomorphic bacteria, which can remain alive for a long time in extreme environmental conditions and as an opportunistic pathogen in humans, easily colonize and affecting people with compromised immune systems in hospital, good effects were achieved. It has been demonstrated that PHMB is effectively used on animate and inanimate surfaces that is colonized with multi drug-resistant Acinetobacter for antisepsis and disinfection.

(125) Bactericidal and Yeasticidal Efficacy of the Formulations A and B

(126) General Description of the Test Method for Bactericidal Efficacy:

(127) The bactericidal activity of a solution containing PHMB and poloxamer 188 according to the present invention was evaluated in accordance with the European Standard EN 13727 (2012/FprA1:2013).

(128) A suspension of test organisms in a solution of the interfering substance is added to a sample of the solution according to the present invention (diluted with hard water). The mixture is maintained at 201 C. for the required contact times. At the end of the contact time, an aliquot of 1 ml is taken; the microbicidal activity in this portion is immediately neutralized. Two 1 ml samples (per dilution step, diluent containing neutralizer) of this suspension are spread on at least 2 plates each. The number of surviving test organisms in the test mixture is calculated for each sample and the reduction is determined with respect to the corresponding test suspension No. The experimental conditions (control A), the non-toxicity of the neutralizer (control B) and the dilution-neutralization method (control C) are validated.

(129) The formulation in form of a solution used contains 0.1 g PHMB, 1.0 g Poloxamer 188 and 98.9 g distilled water per 100 g.

(130) The formulation in form of a gel used contains 0.1 g PHMB, 1.0 g poloxamer 188, 8.6 g glycerin, 1.8 g hydroxyethyl cellulose and 88.5 g distilled water per 100 g.

(131) The test with the solution according to the present invention was performed under dirty conditions (0.3% albumin +0.3% sheep erythrocytes) using Psuedomonas aeruginosa (ATCC 15442), Escherichia coli (NCTC 10538), Enterococcus hirae (ATCC 10541) and Staphylococcus aureus (ATCC 6538), as test-organisms. 80%, 50% and 25% test solutions of the formulation containing 0,1% PHMB (CAS Nr.: 32289-58-0) and 1% Poloxamer (CAS Nr.: 9003-11-6) were used. The pH-values were 6.21 (100%), 7,36 (80%), 7,35 (50%) and 6.88 (WSH). As neutralizer, 4% Tween80+3% Saponin+0.4% Lecithin+0.5% SDS (Neutralizer XXIII) were used. Results are presented below.

(132) Furthermore the solution is tested against MRSA (ATCC 33592), VRE (DSM 17050) and Acinetobacter baumannii l(ATCC 19606). The test performed under dirty (MRSA and VRE) as decribed above and clean conditions (0.3g/l bovine albumin). Results are presented below.

(133) The test with the gel according to the present invention was performed under dirty conditions (0.3% albumin +0.3% sheep erythrocytes) using Psuedomonas aeruginosa (ATCC 15442), Escherichia coli (NCTC 10538), Enterococcus hirae (ATCC 10541) and Staphylococcus aureus (ATCC 6538) as test-organisms. 80%, 50%, 25% and 5% test solutions of the formulation containing 0,1% PHMB (CAS Nr.: 32289-58-0) and 1% Poloxamer (CAS Nr.: 9003-11-6) were used. The pH-values were 6.20 (80%), 7.19 (50%), 7.22 (25%), 7.68 (5%) and 7.05 (WSH). As neutralizer, 4% Tween80+3% Saponin+0.4% Lecithin+0.5% SDS (Neutralizer XXIII) were used. Results are presented below.

(134) Results for Bactericidal Activity:

(135) The solution according to the present invention possesses bactericidal activity (log.sub.10 RF5) at 20 C. under dirty conditions for reference strains P. aeruginosa, E. coli, S. aureus and E. hirae within a contact time of 1 min at a product concentration of 80%, and within a contact time of 5 min at a product concentration of 25%. Furthermore, the said solution possesses a bactericidal activity (log.sub.10 RF5) at 20 C. under dirty conditions in 5 and 10 minutes when diluted at 80% and 50% (v/v) in distilled water for the reference strain Staphylococcus aureus (MRSA) and in 1, 5 and 10 minutes when diluted at 80% and 50%(v/v) in distilled water for the reference strain Enterococcus faecium (VRE). The test result at 20 C. under clean conditions for the reference strain Acinetobacter baumannii showed that the solution possesses a bactericidal activity (log.sub.10 RF 5) in 5 and 10 minutes when diluted at 80% and 50% (v/v) in distilled water.

(136) The gel according to the present invention possesses bactericidal activity (log.sub.10 RF5) at 20 C. under dirty conditions for reference strains P. aeruginosa, E. coli, S. aureus and E. hirae within a contact time of 1 min at a product concentration of 50%.

(137) General Description of the Test Method for Yeasticidal Efficacy:

(138) Die yeasticidal activity of a solution containing PHMB and poloxamer 188 according to the present invention was evaluated in accordance with the European Standard EN 13624 (2013).

(139) A suspension of test organisms in a solution of the interfering substance is added to a sample of the solution according to the present invention (diluted with hard water). The mixture is maintained at 201 C. for the required contact times. At the end of the contact time, an aliquot of 1 ml is taken; the microbicidal activity in this portion is immediately neutralized. Two 1 ml samples (per dilution step, diluent containing neutralizer) of this suspension are spread on at least 2 plates each. The number of surviving test organisms in the test mixture is calculated for each sample and the reduction is determined with respect to the corresponding test suspension No. The experimental conditions (control A), the non-toxicity of the neutralizer (control B) and the dilution-neutralization method (control C) are validated.

(140) The formulation in form of a solution used contains 0.1 g PHMB, 1.0 g Poloxamer 188 and 98.9 g distilled water per 100 g.

(141) The formulation in form of a gel used contains 0.1 g PHMB, 1.0 g poloxamer 188, 8.6 g glycerin, 1.8 g hydroxyethyl cellulose and 88.5 g distilled water per 100 g.

(142) The test with the solution according to the present invention was performed under dirty conditions (0.3% albumin+0.3% sheep erythrocytes) using C. albicans (ATCC 10231) as test-organism. 80%, 50% and 25% test solutions of the formulation containing 0.1% PHMB (CAS Nr.: 32289-58-0) and 1% Poloxamer (CAS Nr.: 9003-11-6) were used. The pH-values were 6.21 (100%), 7.36 (80%), 7.35 (50%) and 6.88 (WSH). As neutralizer, 4% Tween80+3% Saponin+0.4% Lecithin+0.5% SDS (Neutralizer XXIII) were used. Results are presented below.

(143) The test with the gel according to the present invention was performed under dirty conditions (0.3% albumin+0.3% sheep erythrocytes) using C. albicans (ATCC 10231) as test-organism. 80%, 50%, 25% and 5% test solutions of the formulation containing 0.1% PHMB (CAS Nr.: 32289-58-0) and 1% Poloxamer (CAS Nr.: 9003-11-6) were used. The pH-values were 6.20 (80%), 7.19 (50%), 7.22 (25%), 7.68 (5%) and 7.05 (WSH). As neutralizer, 4% Tween80+3% Saponin+0.4% Lecithin+0.5% SDS (Neutralizer XXIII) were used. Results are presented below.

(144) Results for Yeasticidal Activity:

(145) The solution according to the present invention possesses bactericidal activity (log.sub.10 RF4) at 20 C. under dirty conditions for reference strain C. albicans within a contact time of 15 min at a product concentration starting at 50%.

(146) The gel according to the present invention possesses bactericidal activity (log.sub.10 RF4) at 20 C. under dirty conditions for reference strain C. albicans within a contact time of 15 min at a product concentration starting at 25%.

(147) Comparison with Other Formulations

(148) Bactericidal and Yeasticidal Activity

(149) The formulations of the present invention have the same or better bactericidal and/or yeasticidal activity as commonly used formulations.

(150) For instance, the solution according to the present invention containing PHMB and poloxamer (e.g. poloxamer 188) has the same or a better bactericidal and yeasticidal activity compared to PHMB used alone.

(151) Further, the solution according to the present invention containing PHMB and poloxamer (e.g. poloxamer 188) has the same or a better bactericidal and yeasticidal activity compared to PHMB in combination with undecylenamidopropyl betaine.

(152) Further, the solution according to the present invention containing PHMB, poloxamer (e.g. poloxamer 188) and a co-surfactant or a mixture of co-surfactants has the same or a better bactericidal and yeasticidal activity compared to PHMB in combination with undecylenamidopropyl betaine or poloxamer.

(153) Further, the gel according to the present invention containing PHMB, poloxamer (e.g. poloxamer 188), glycerin and hydroxyethylcellulose has the same or a better bactericidal and yeasticidal activity compared to PHMB used alone with glycerin and hydroxyethyl cellulose.

(154) Further, the gel according to the present invention containing PHMB, poloxamer (e.g. poloxamer 188), glycerin and hydroxyethyl cellulose has the same or a better bactericidal and yeasticidal activity compared to PHMB used with glycerin and hydroxyethyl cellulose and undecylenamidopropyl betaine.

(155) Further, the gel according to the present invention containing PHMB, poloxamer (e.g. poloxamer 188), glycerin, hydroxyethyl cellulose and a co-surfactant or a mixture of co-surfactants has the same or a better bactericidal and yeasticidal activity compared to PHMB used with undecylenamidopropyl betaine or poloxamer and glycerin and hydroxyethyl cellulose.

(156) Further, the solution according to the present invention containing PHMB, poloxamer (e.g. poloxamer 188), a substance/substances obtained from herbs and optionally a co-surfactant or a mixture of co-surfactants has the same or a better bactericidal and yeasticidal activity compared to a solution not containing said substance/substances obtained from herbs.

(157) Further, the gel according to the present invention containing PHMB, poloxamer (e.g. poloxamer 188), glycerin, hydroxyethyl cellulose, a substance/substances obtained from herbs and optionally a co-surfactant or a mixture of co-surfactants has the same or a better bactericidal and yeasticidal activity compared to a solution not containing said substance/substances obtained from herbs.

(158) Toxicity

(159) The composition with PHMB and poloxamer has improved properties compared to other surfactants such as betaine-derivatives. It has turned out that poloxamers and in particular poloxamer 188 have a significantly lower toxicity than most commonly used surfactants such as betaine-derivatives. Classification information according to EU regulations and LD50 values of some commercial surfactants are given in table 5.

(160) TABLE-US-00031 TABLE 5 Comparison of hazardous information of Kolliphor P 188 (Poloxamer) with various commercially available unbranched fatty acid derivative surfactants. Cas GHS LD50 Acute Chemical name Number Classification.sup.a oral (Rat) Kolliphor P 188 9003-11-6 No hazard warning >5.000 mg/kg (Poloxamer 188) label required CRODATERIC CAB 30-LQ-(RB) 61789-40-0 Eye Irrit. 2; H319 no data available (Cocamidopropyl Betaine) CRODATERIC CAB 30-LQ-(TH) 4292-10-8 Skin Irrit. 2; H315 >4.900 mg/kg (Lauramidopropyl betaine) Eye Irrit. 2; H319 Rewoteric AM B U 185 98510-75-9 Eye Dam. 1; H318 >2.000 mg/kg (Undecylenamidopropyl Betaine) .sup.aCLP-Regulation (EC) No 1272/2008

(161) As mentioned above, the cytotoxicity is an important key in wound recovery. Further the cytotoxicity reducing effect of an ingredient is a desirable. It has been shown that poloxamer 188 has membrane sealing and repairing properties on neurons, cardiac myocytes, fibroblasts, and skeletal muscle cells [22-25]. Thus poloxamer 188 prevents apoptosis and reduce cytotoxicity. Additionally, it was reported that poloxamer 188 facilitates plasma membrane repair in alveolus resident cells [26].

(162) Thus, it is beneficial to use poloxamers instead of commonly used surfactants in bactericidal or yeasticidal formulations.

(163) Surface Tension

(164) Furthermore, tri-blockcopolymers of the present invention, in particular poloxamers, due to their non-ionic properties have improved surface tension compared to surfactants such as betaine-derivatives. For instance, in case the pH on a wound changes, the charge of the betaines may change and thus have a different surface tension than before. Therefore, with betaine-derivatives, the desired bactericidal or yeasticidal effect may not always be reproducibly ensured. Tri-blockcopolymers of the present invention on the other hand have the same surface tension, independent of the surrounding pH and ensure a constant and reliable bactericidal or yeasticidal effect.

(165) At the same time, the adhesion of microorganisms on wounds is minimized better by said tri-blockcopolymers.

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