ANTIMICROBIAL PRESERVATIVE COMPOSITIONS

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent Application No. 62/293,840, filed

February 11, 2016, the contents of which are incorporated herein by reference in their entirety.

FIELD OF INVENTION

The present invention, in some embodiments thereof, relates to personal care formulations and, more particularly, but not exclusively, to stable personal care formulations with antimicrobial activity, articles containing same, and uses thereof in, for example, reducing or preventing growth of microorganisms .

BACKGROUND OF THE INVENTION

Antimicrobial compositions are used, for example, in the health care industry, food service industry, meat processing industry, and in the private sector by individual consumers.

Preservation of personal care products from microbial contamination has become a difficult task since the available approved antimicrobials are very few and those which have good antimicrobial activity are quite toxic. Consumers seek for products for topical applications being free from toxic antimicrobials that can be used as preservatives.

Infection is a constant risk to any healthy person, and poses even a higher risk to hospitalized patients. The risk of infection is further increased when the natural infection barriers of skin or other epithelial surfaces are breached during a surgical procedure, and/or otherwise in cases where bacteria normally present on the skin or in the air are allowed to access the interior surfaces of the body.

Hospital-acquired (nosocomial) infections occur even when good hygiene and sterile technique is followed, although the incidence is reduced. Despite even the most rigorous aseptic procedures, people cannot be completely isolated from infectious agents.

The widespread use of antibacterial compositions indicates the importance consumers place on controlling bacteria and other microorganism populations on skin. It is important, however, that antibacterial compositions provide a substantial and broad spectrum reduction in microorganism populations quickly and without problems associated with toxicity and skin irritation. In particular, antibacterial cleansing compositions typically contain an active antibacterial agent, a surfactant, and various other ingredients, for example, dyes, fragrances, pH adjusters, thickeners, and the like, in an aqueous carrier.

Despite being successful in controlling or eliminating bacterial infections, widespread use of antibiotics both in human medicine and as a feed supplement in poultry and livestock production has led to drug resistance in many pathogenic bacteria (McCormick J. B., Curr Opin Microbiol 1: 125- 129, 1998). The evolution and spread of resistance genetic determinants, multidrug resistant (MDR) bacteria that cause life-threatening infections have been increasingly emerged (A. P. Magiorakos et al. Clin. Microbiol. Infect. 2012, 18, 268), and as such, the effectiveness of antibiotics has greatly diminished in the last decade. Furthermore, as resistance spreads among bacteria, there is great concern that antibiotics treatment will become increasingly less effective and, in some cases, completely ineffective.

Nosocomial infections caused by antibiotic -resistant bacteria result in patient suffer and mortality and impose a substantial burden on the medical system due to extended periods of hospitalization. The economic impact of managing infections caused by nosocomial infections is substantial, and costs are estimated to be more than $4 billion annually.

Currently, a number of blends are available for preservation of personal care products where synergy between antimicrobials is exploited in lowering the concentration of individual ingredient. Another great advantage is that the microbes cannot develop resistance very easily if they are attacked by a combination of antimicrobials.

European patent 2,209,392 discloses preservatives and antimicrobial agents comprising extracts derived from sugar cane.

SUMMARY OF THE INVENTION

The present inventors have surprisingly uncovered that personal care formulations comprising thymol and linalool, phenoxyethanol or phenyl ethyl alcohol at specified ratios, can be readily prepared, and that such formulations exhibit exceptional and even synergistic antimicrobial activity.

According to one aspect, the present invention provides a formulation comprising: thymol, an organic solvent and an odor masking agent selected from the group consisting of: linalool and liffarome. In some embodiments, the thymol and organic solvent are in a ratio that ranges from 1: 1 to 15:85, by weight, respectively. In some embodiments, the thymol and odor masking agent are in a ratio that ranges from 100:0.1 to 1: 1, by weight, respectively. In some embodiments, the thymol and linalool are in a ratio that ranges from 95:5 to 1: 1, by weight, respectively. In some embodiments, the the thymol and liffarome are in a ratio that ranges from 100:0.1 to 4: 1, by weight, respectively.

In some embodiments, the organic solvent is selected from the group consisting of: propylene glycol, benzyl alcohol, phenyl ethyl alcohol, phenoxyethanol, each being substituted or non- substituted, or a mixture thereof. In some embodiments, said organic solvent is effective as an antibacterial agent. In some embodiments, said organic solvent is an alcohol.

In some embodiments, the organic solvent is phenyl ethyl alcohol. In some embodiments, the organic solvent is phenoxyethanol. In some embodiments, the organic solvent is benzyl alcohol.

In some embodiments, the formulation comprises liffarome, and the organic solvent is selected from the group consisting of: phenoxyethanol or phenyl ethyl alcohol. In some embodiments, the formulation comprises liffarome and further comprises capric acid.

In some embodiments, the formulation comprises: 15 - 35 % (w/w) thymol; 3 - 20% linalool

(w/w); and 30% - 80% (w/w) organic solvent.

In some embodiments, the organic solvent ranges from 40% to 80%, by weight, respectively.

In some embodiments, the thymol and linalool are in a ratio that ranges from 80:20 to 60:40, by weight.

In some embodiments, the formulation comprises: 15 - 35 % (w/w) thymol; 65- 85 % (w/w) phenoxyethanol or phenyl ethyl alcohol; and 0.1 - 3 % (w/w) liffarome. In some embodiments, the formulation further comprises 1 -5 % (w/w) capric acid.

In some embodiments, the phenoxyethanol or phenyl ethyl alcohol range from 40% to 80%, by weight, respectively.

In some embodiments, the thymol and phenoxyethanol or phenyl ethyl alcohol are in a ratio that ranges from 80:20 to 60:40, by weight.

In some embodiments, the formulation is a synergistic antimicrobial formulation.

In some embodiments, the formulation is for use in the treatment of a medical, cosmetic and/or cosmeceutical condition.

According to one aspect, the present invention provides an article comprising the disclosed formulation. In some embodiments, the article is a personal care product. In some embodiments, the product is or comprises a formulation in the form selected from the group consisting of: paste, cream, lotion, foam, gel, emulsion, an ointment, and soap.

In some embodiments, the article is selected from the group consisting of: a fabric, a bandage, a wipe, a pledget, a swab, a suppository, a dressing, a solution, a mousse, a pad, and a patch.

In some embodiments, the article is identified for use in the treatment of a condition selected from medical, cosmetic and cosmeceutical condition.

According to one aspect, the present invention provides a method of inhibiting or reducing the formation of load of a microorganism in and/or on an article, the method comprising contacting said article with the disclosed formulation. In some embodiments, the microorganism is selected from bacteria, molds and fungi. In some embodiments, the bacteria are Gram positive bacteria selected from the group consisting of: Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus cereus; or Gram negative bacteria selected from the group consisting of: Escherichia coli, Pseudomonas aeuruginosa, and Burkholderia cepacia. In some embodiments, the fungi are selected from the group consisting of: Candida albicans. In some embodiments, the mold is Aspergillus niger.

Further embodiments and the full scope of applicability of the present invention will become apparent from the detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

DETAILED DESCRIPTION OF THE INVENTION

The present invention, in some embodiments thereof, relates to compositions and, more particularly, but not exclusively, to personal care formulations with antimicrobial (also referred to as "anti-micro-organic") activity, articles containing same, and uses thereof in, for example, reducing or preventing growth of microorganisms.

Before explaining at least one embodiment of the invention in detail, it is to be understood that the invention is not necessarily limited in its application to the details set forth in the following description or exemplified by the Examples. The invention is capable of other embodiments or of being practiced or carried out in various ways. High quality antimicrobial formulations are desirable as they provide a good solution to biofouling and/or infection processes and/or formation of biofilms on a surface. Conventional personal care formulations present several drawbacks, as many of these formulations are referred to as being toxic (or releasing toxic materials to the environment), instable, inefficient or are limited in preventing (or complete diminishing) microorganism growth, expensive and produced via complicated manufacturing processes which at times require expensive equipment for their manufacture.

The present inventors have designed non-toxic antimicrobial formulations (also referred to as "blends") comprising active compounds of natural extracts e.g., thymol, and linalool, as well as phenoxyethanol or phenyl ethyl alcohol in a weight ratio of 9: 1 to 5:5, respectively, for use as e.g., cosmetic composition and for various articles. Non -limiting examples of such uses can include personal hygiene and treatment of specific skin regions-of-interest. In some embodiments, the disclosed formulations are both efficient, cost-effective antimicrobial formulations.

In some embodiments, "linalool" refers to (R)-(-)-linalool (also known as licareol).

In some embodiments, "linalool" refers to (S)-(+)-linalool (also known as coriandrol). In some embodiments, "linalool" refers to a mixture of licareol and coriandrol.

As demonstrated in the Examples section that follows, the present inventors have shown that compositions (e.g., formulations) comprising thymol and linalool, being at certain predetermined ratios, and a solvent (e.g., propylene glycol (designated as PG) or phenyl ethyl alcohol), or compositions comprising thymol and phenoxyethanol, or phenyl ethyl alcohol being at certain predetermined ratios, exhibit a desired stability as well as improved antimicrobial activities, compared to other compositions comprising different materials from the invented formulations or from formulations comprising the same materials but having different volume or weight ratios thereof.

In exemplary embodiments, the desired solvents are selected from the group consisting of: phenoxyethanol, benzyl alcohol, and phenyl ethanol or a combination thereof.

In some embodiments, the solvents are devoid of 1 ,2-propylene glycol and/or hexylene glycol. As demonstrated in the Examples section that follows, the present inventors have shown that a specific ratio of thymol/linalool exhibits both synergistic antimicrobial effect and further an effective odor control benefit, i.e. masking the odor of thymol. Similarly, a specific ratio of thymol/ phenoxyethanol, or phenyl ethyl alcohol with the addition of odor masking agents exhibits similar effects. In some embodiments, the odor masking agent are selected from: liffarome, capric acid, or a combination thereof.

In some embodiments, "liffarome" refers to cis-hex-3-enyl methyl carbonate. In some embodiments, "liffarome" refers to the chemical with CAS no. 67633-96-9. In some embodiments, "liffarome" refers to liffarome and any derivatives thereof. In some embodiments, liffarome further refers to any enantiomer, diastereomer, solvate or hydrate of liffarome.

In some embodiments, "capric acid" refers to decanoic acid. In some embodiments, "capric acid" refers to the chemical with CAS no. 334-48-5. In some embodiments, "capric acid" refers to capric acid and any derivatives thereof. In some embodiments, capric acid further refers to any enantiomer, diastereomer, solvate or hydrate of capric acid.

In some embodiments, the compounds described herein above possess asymmetric carbon atoms (optical centers) or double bonds; the racemates, diastereomers, geometric isomers and individual isomers are encompassed within the scope of the present invention.

As used herein and in the art, the term "enantiomer" describes a stereoisomer of a compound that is superposable with respect to its counterpart only by a complete inversion/reflection (mirror image) of each other. Enantiomers are said to have "handedness" since they refer to each other like the right and left hand. Enantiomers have identical chemical and physical properties except when present in an environment which by itself has handedness, such as all living systems.

In some embodiments, the compounds hereabove described can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are encompassed within the scope of the present invention.

The term "solvate" refers to a complex of variable stoichiometry (e.g., di-, tri-, tetra-, penta-, hexa-, and so on), which is formed by a solute (the conjugate described herein) and a solvent, whereby the solvent does not interfere with the biological activity of the solute. Suitable solvents include, for example, ethanol, acetic acid and the like.

The term "hydrate" refers to a solvate, as defined hereinabove, where the solvent is water. The present inventors have also shown that the disclosed formulation can be used to impart to articles the antimicrobial activities.

The formulation:

According to an aspect of some embodiments of the present invention there is provided a stable composition. In some embodiments, the composition is in the form of a formulation. In some embodiments, the composition is a stable preservative. In some embodiments, the stable personal care formulation comprises: thymol (also known as 2-isopropyl-5-methylphenol), an organic solvent and Hnalool (also known as 3,7-dimethylocta-l,6-dien-3-ol). In some embodiments, the thymol and Hnalool are at a ratio of e.g., about 9: 1, about 8:2, about 7:3, about 7:2, 6:4, 5:5, respectively, by weight, including any value and range therebetween.

In some embodiments, the stable personal care formulation comprises: thymol and phenoxyethanol (also known as l-hydroxy-2-phenoxyethane). In some embodiments, the thymol and phenoxyethanol are at a ratio of e.g., about 9: 1, about 8:2, about 7:3, about 7:2, 6:4, 5:5, respectively, by weight, including any value and range therebetween. In some embodiments, the stable personal care formulation comprises: thymol and phenyl ethyl alcohol (also known as 2-phenylethanol). In some embodiments, the thymol and phenyl ethyl alcohol are at a ratio of e.g., about 9: 1, about 8:2, about 7:3, about 7:2, 6:4, 5:5, respectively, by weight, including any value and range therebetween.

In some embodiments of the present invention, the formulation is a personal care formulation.

In some embodiments of the present invention, the formulation comprises thymol at a concentration of e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, by weight, including any value and range therebetween.

In some embodiments of the present invention, the formulation comprises Hnalool at a concentration of e.g., about 3%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, by weight, including any value and range therebetween.

In some embodiments of the present invention, the formulation comprises phenoxyethanol at a concentration of e.g., about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, by weight, about 90%, by weight, including any value and range therebetween.

In some embodiments of the present invention, the formulation comprises phenyl ethyl alcohol at a concentration of e.g., about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, by weight, about 90%, by weight, including any value and range therebetween.

In some embodiments of the present invention, the formulation further comprises a solvent. In exemplary embodiments, the formulation comprises 65% solvent (by weight) and further comprises:

(a) 2.5% thymol and 31.5% Hnalool, or (b) 7.0% thymol and 28% linalool or (c) 10.5% thymol and 24.5% linalool or (d) 14% thymol and 21% linalool or (e) 17.5% thymol and 17.5% linalool or (f) 21% thymol and 14% linalool or (g) 24.5% thymol and 10.5% linalool or (h) 28% thymol and 7% linalool or (i) 28% thymol and 7% linalool. The percent of the solvent and of thymol and linalool in (a)-(i) may vary within ± 20%.

Embodiments of the solvent are described hereinbelow.

In some embodiments, the solvent comprises 0-10% water.

In some embodiments, the organic solvent comprises diol or triol.

In some embodiments, the glycol is selected from the group consisting of ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, pentylene glycol and hexylene glycol.

In some embodiments, the organic solvent comprises benzyl alcohol, phenoxyethanol, phenyl ethyl alcohol, 3-phenyl propanol, substituted or non-substituted, or a combination thereof.

In some embodiments of the present invention, the formulation further comprises an odor masking agent. In some embodiments, the odor masking agent is liffarome. In some embodiments, the odor masking agent is capric acid. In some embodiments, the odor masking agent is a combination of liffarome and capric acid.

In exemplary embodiments, the formulation comprises 0.12% liffarome (by weight) and further comprises:

(a) 2.5% thymol and 97.38% phenoxyethanol or phenyl ethyl alcohol, or (b) 7.0% thymol and

92.88% phenoxyethanol or phenyl ethyl alcohol or (c) 10.5% thymol and 89.38% phenoxyethanol or phenyl ethyl alcohol or (d) 14% thymol and 85.88% phenoxyethanol or phenyl ethyl alcohol or (e) 17.5% thymol and 82.38% phenoxyethanol or phenyl ethyl alcohol or (f) 21% thymol and 78.88% phenoxyethanol or phenyl ethyl alcohol or (g) 24.5% thymol and 75.38% phenoxyethanol or phenyl ethyl alcohol or (h) 28% thymol and 71.88% phenoxyethanol or phenyl ethyl alcohol or (i) 31.5% thymol and 68.38% phenoxyethanol or phenyl ethyl alcohol or (j) 35% thymol and 64.88% phenoxyethanol or phenyl ethyl alcohol.

In exemplary embodiments, the formulation comprises 0.12% liffarome (by weight) and 1.5% capric acid (by weight) and further comprises:

(a) 2.5% thymol and 95.88% phenoxyethanol or phenyl ethyl alcohol, or (b) 7.0% thymol and

91.38% phenoxyethanol or phenyl ethyl alcohol or (c) 10.5% thymol and 87.88% phenoxyethanol or phenyl ethyl alcohol or (d) 14% thymol and 84.38% phenoxyethanol or phenyl ethyl alcohol or (e) 17.5% thymol and 80.88% phenoxyethanol or phenyl ethyl alcohol or (f) 21% thymol and 77.38% phenoxyethanol or phenyl ethyl alcohol or (g) 24.5% thymol and 73.88% phenoxyethanol or phenyl ethyl alcohol or (h) 28% thymol and 70.38% phenoxyethanol or phenyl ethyl alcohol or (i) 31.5% thymol and 66.88% phenoxyethanol or phenyl ethyl alcohol or (j) 35% thymol and 63.88% phenoxyethanol or phenyl ethyl alcohol.

In some embodiments, liffarome is at a concentration of e.g., about 0.05 %, 0.12 %, 0.22 %, 0.32 %, 0.42 %, 0.52 %, 0.62 %, 0.72 %, 0.82 %, 0.92 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, or 10 %, by weight, including any value and range therebetween.

In some embodiments, capric acid is at a concentration of e.g., about 0.05 %, 0.1 %, 0.5 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 15 %, or 20 %, by weight, including any value and range therebetween.

The formulation of the invention may include an emulsifier.

As used herein and in the art, the term "emulsifier" is intended to mean a surface-active agent that facilitates the mixing of two or more liquid substances that would separate into its component parts under normal conditions.

Surface- active agents may include surfactants, which typically provide detersive functionality to a formulation or act simply as wetting agents. Surface-active agents may generally be categorized as anionic surface-active agents, cationic surface-active agents, nonionic surface-active agents, amphoteric surface-active agents and zwitterionic surface-active agents, and dispersion polymers.

Exemplary emulsifiers include, but are not limited to, sodium lauroyl lactylate (SLL), Tween 20, PEG 40-hdrogenatedcster oil, cocoamide monoethanolamide (MEA), cocoamide diethanolamid (DEA), diacetyl tartaric acid ester of mono- and diglycerides (DATEM), potassium cocoate, and any combination thereof.

In some embodiments, the emulsifier is at a concentration of e.g., about 0.05 %, 0.1 %, 0.2 %, 0.3 %, 0.4 %, 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 15 %, or 20 %, by weight, including any value and range therebetween.

In some embodiments, the formulation is devoid of paraben. In some embodiments, the formulation is devoid of formaldehyde. In some embodiments, the formulation is devoid of a compound comprising halogen. In some embodiments, the formulation is devoid of a compound comprising halogen and is further devoid of paraben. In some embodiments, the formulation is devoid of formaldehyde and is further devoid of paraben. In some embodiments, the formulation is devoid of a compound comprising halogen and is further devoid of paraben. In some embodiments, the formulation is devoid a compound comprising halogen and is further devoid of formaldehyde. In some embodiments, the formulation is devoid of paraben, formaldehyde and is further devoid of a compound comprising halogen

In some embodiments, the formulation is not pH dependent.

In some embodiments of the present invention, the formulation is characterized by pH below 7. In some embodiments of the present invention, the formulation is characterized by pH below 6. In some embodiments of the present invention, the formulation is characterized by pH below 5. In some embodiments of the present invention, the formulation is characterized by pH below 4. In some embodiments of the present invention, the formulation is characterized by pH below 3.

In some embodiments of the present invention, the formulation is characterized by pH in the ranges of about 5 to about 7. In some embodiments of the present invention, the formulation is a part of a product, article or composition characterized by pH in the ranges of about 3 to about 9. In some embodiments of the present invention, the formulation further comprises a buffer solution or a pH adjuster to control the desired pH of the formulation.

As used herein, the terms "formulation", or "blend", which are used hereinthroughout interchangeably, refer to a vehicle composition in the form of emulsion, lotion, cream, gel etc., that optionally further comprises physiologically acceptable carriers and/or excipients and optionally other chemical components such as cosmetically, cosmeceutically or pharmaceutically active agents (e.g., drugs). The formulation can optionally further comprise a carrier, and optionally additional active agents and/or additives e.g., anti-freezing agents).

The term "by weight," unless otherwise specified, means by weight of the total composition.

As used herein, the term "physiologically acceptable" means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

Herein the term "excipient" refers to an inert substance added to a formulation as described herein to further facilitate processes and administration of the active ingredients.

The formulation of the invention can be prepared by any commonly used method for preparing a composition of materials. For example, the components of the formulations may be added and mixed together, or one of the components may be added to the other in the form of a solution which may, if desired, be evaporated or lyophilized after mixing for obtaining a homogeneous and stable solution or suspension. As used herein the terms "stable formulation", or "long-lasting formulation", mean that the formulation remains in a state or condition of sufficient stability to have utility as a personal care agent, while maintaining the antimicrobial activity (with ± 20% variation). For example, and without limitation, the formulation has a sufficient stability to allow storage at a convenient temperature, e.g., between 10 °C and 30 °C, for a reasonable period of time of e.g., longer than one month, longer than three months, longer than six months, and longer than one year.

As used herein, the phrase "personal care" refers to compositions that can be formulated in various cosmetic and pharmaceutical consumer products utilizing a variety of delivery systems and carrier bases. Such consumer product forms include, but are not limited to, shampoos, aftershaves, sunscreens, body and hand lotions, skin creams, liquid soaps, bar soaps, bath oil bars, shaving creams, conditioners, permanent waves, hair relaxers, hair bleaches, hair detangling lotion, styling gel, styling glazes, spray foams, styling creams, styling waxes, styling lotions, mousses, spray gels, pomades, shower gels, bubble baths, hair coloring preparations, conditioners, hair lighteners, coloring and non- coloring hair rinses, hair grooming aids, hair tonics, spritzes, styling waxes, band-aids, and balms.

In some embodiments, the disclosed formulation is in the form of, or a part of, a cream, an ointment, a paste, a gel, a lotion, a milk, an oil, a suspension, a solution, an aerosol, a spray, a foam, or a mousse.

Ointments are semisolid preparations, typically based on petrolatum or petroleum derivatives. The specific ointment base to be used is one that provides for optimum delivery for the active agent chosen for a given formulation, and, preferably, provides for other desired characteristics as well (e.g., emolliency). As with other carriers or vehicles, an ointment base should be inert, stable, nonirritating and nonsensitizing. As explained in Remington: The Science and Practice of Pharmacy, 19th Ed., Easton, Pa.: Mack Publishing Co. (1995), pp. 1399-1404, ointment bases may be grouped in four classes: oleaginous bases; emulsifiable bases; emulsion bases; and water-soluble bases. Oleaginous ointment bases include, for example, vegetable oils, fats obtained from animals, and semisolid hydrocarbons obtained from petroleum. Emulsifiable ointment bases, also known as absorbent ointment bases, contain little or no water and include, for example, hydroxystearin sulfate, anhydrous lanolin and hydrophilic petrolatum. Emulsion ointment bases are either water-in-oil (W/O) emulsions or oil-in-water (O/W) emulsions, and include, for example, cetyl alcohol, glyceryl monostearate, lanolin and stearic acid. Exemplary water-soluble ointment bases are prepared from polyethylene glycols of varying molecular weight. Lotions are preparations that are to be applied to the skin surface without friction. Lotions are typically liquid or semiliquid preparations in which solid particles, including the sunscreens- containing microcapsules, are present in a water or alcohol base. Lotions are typically preferred for covering/protecting large body areas, due to the ease of applying a more fluid composition. Lotions are typically suspensions of solids, and oftentimes comprise a liquid oily emulsion of the oil-in-water type. It is generally necessary that the insoluble matter in a lotion be finely divided. Lotions typically contain suspending agents to produce better dispersions as well as compounds useful for localizing and holding the active agent in contact with the skin, such as methylcellulose, sodium carboxymethyl- cellulose, and the like.

Creams are viscous liquids or semisolid emulsions, either O/W or W/O. Cream bases are typically water- washable, and contain an oil phase, an emulsifier and an aqueous phase. The oil phase, also called the "internal" phase, generally comprises petrolatum and/or a fatty alcohol such as cetyl or stearyl alcohol. The aqueous phase typically, although not necessarily, exceeds the oil phase in volume, and generally contains a humectant. The emulsifier in a cream formulation is generally a nonionic, anionic, cationic or amphoteric surfactant. Reference may be made to Remington: The Science and Practice of Pharmacy, supra, for further information.

Pastes are semisolid dosage forms in which the bioactive agent is suspended in a suitable base. Depending on the nature of the base, pastes are divided between fatty pastes or those made from a single-phase aqueous gel. The base in a fatty paste is generally petrolatum, hydrophilic petrolatum and the like. The pastes made from single-phase aqueous gels generally incorporate carboxymethylcellulose or the like as a base. Additional reference may be made to Remington: The Science and Practice of Pharmacy, for further information.

Gel formulations are semisolid, suspension-type systems. Single-phase gels contain organic macromolecules distributed substantially uniformly throughout the carrier liquid, which is typically aqueous, but also, preferably, contain an alcohol and, optionally, an oil. Preferred organic macromolecules, i.e. gelling agents, are crosslinked acrylic acid polymers such as the family of carbomer polymers, e.g., carboxypolyalkylenes that may be obtained commercially under the trademark Carbopol™. Other types of preferred polymers in this context are hydrophilic polymers such as polyethylene oxides, polyoxyethylene-polyoxypropylene copolymers and polyvinylalcohol; cellulosic polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, and methyl cellulose; gums such as tragacanth and xanthan gum; sodium alginate; and gelatin. In order to prepare a uniform gel, dispersing agents such as alcohol or glycerin can be added, or the gelling agent can be dispersed by trituration, mechanical mixing or stirring, or combinations thereof.

Sprays generally provide the active agent in an aqueous and/or alcoholic solution which can be misted onto the skin for delivery. Such sprays include those formulated to provide for concentration of the active agent solution at the site of administration following delivery, e.g., the spray solution can be primarily composed of alcohol or other like volatile liquid in which the active agent can be dissolved. Upon delivery to the skin, the carrier evaporates, leaving concentrated active agent at the site of administration.

Foam compositions are typically formulated in a single or multiple phase liquid form and housed in a suitable container, optionally together with a propellant which facilitates the expulsion of the composition from the container, thus transforming it into a foam upon application. Other foam forming techniques include, for example the "Bag-in-a-can" formulation technique. Compositions thus formulated typically contain a low-boiling hydrocarbon, e.g., isopropane. Application and agitation of such a composition at the body temperature cause the isopropane to vaporize and generate the foam, in a manner similar to a pressurized aerosol foaming system. Foams can be water-based or hydroalcoholic, but are typically formulated with high alcohol content which, upon application to the skin of a user, quickly evaporates, driving the active ingredient through the upper skin layers to the site of treatment.

Personal care formulations can further include, without limitation, human body or hair deodorizing solution, deodorizing gel, deodorizing spray, deodorizing stick, deodorizing roll-on, deodorizing paste, deodorizing cream, deodorizing lotion, deodorizing aerosol, and other commonly marketed human body and commonly marketed animal and pet deodorizing compositions.

Additional cosmetically or pharmaceutically beneficial (active) ingredients can also be included in the formulations of the present invention, which can be selected from, but are not limited to, skin cleansers, cationic, anionic surfactants, non-ionic surfactants, amphoteric surfactants, and zwitterionic surfactants, skin and hair conditioning agents, vitamins, hormones, minerals, plant extracts, anti-inflammatory agents, collagen and elastin synthesis boosters, UVA/UVB sunscreens, concentrates of plant extracts, emollients, moisturizers, skin protectants, humectants, silicones, skin soothing ingredients, antimicrobial agents, antifungal agents, treatment of skin infections and lesions, blood microcirculation improvement, skin redness reduction benefits, additional moisture absorbents, analgesics, skin penetration enhancers, solubilizers, moisturizers, emollients, anesthetics, colorants, perfumes, preservatives, seeds, broken seed nut shells, silica, clays, beads, luffa particles, polyethylene balls, mica, pH adjusters, processing aids, and combinations thereof.

In some embodiments of the present invention, the formulation is characterized by resistance to discoloration.

As used herein, "discoloration" is defined as change in the hue or in the visual appearance of a formulation, due to an internal reaction among its constituents or the bleaching or oxidation action caused by a combination of factors that include, but are not limited to, air, high temperature, humidity, ultraviolet exposure e.g., sunlight.

Articles comprising the Formulation:

According to an aspect of some embodiments of the present invention there is provided an article which comprises any one of the personal care formulation described herein.

Some embodiments of this aspect of present embodiments are included hereinabove, under "The Formulation", and form an integral part of embodiments relating to "Articles comprising the Formulation".

According to an aspect of some embodiments of the present invention, there is provided a pharmaceutical, cosmetic or cosmeceutical product comprising the formulation described in any of their respective embodiments herein, for use in treating a medical, cosmetic or cosmeceutic condition, as described herein.

According to an aspect of some embodiments of the present invention, the formulation described in any of their respective embodiments herein is used as, or a part of, a preservative in any pharmaceutical, cosmetic or cosmeceutical product or in any article as describe herein.

As used herein, "preservative" is used to prevent the growth of bacteria, fungi and/or molds in any personal care composition or formulation.

According to a further aspect of some embodiments of the present invention, there is provided a use of the formulation described herein in the manufacture of a pharmaceutical, cosmetic or cosmeceutical product, which can be used in treating a medical, cosmetic or cosmeceutic condition, as described herein.

In some embodiments, there is provided a method of treating a medical, cosmeceutical or cosmetic condition treatable by a topical or transdermal administration, the method comprising topically applying the formulation described herein (e.g., in the context of a pharmaceutical, cosmetic or cosmeceutic product) to a skin or mucosal tissue of a subject afflicted by the condition.

The phrases "topical" "topical administrations" and or any grammatical derivative thereof, is meant to encompass applications, which include, without limitation, dermal applications, ophthalmic application, vaginal application, rectal application and intranasal application.

Medical, cosmetic or cosmeceutical conditions that can benefit from containing the formulations described herein when applied topically, with or without an additional active ingredient, include, but are not limited to, infections caused by pathogenic microorganisms, as discussed in further detail hereinbelow, wounds, particularly when associated with an infection, acne, skin infections, viral blisters such as one caused by herpes, sexual dysfunction such as erectile dysfunction.

Hence, according to some embodiments of the present invention, the pharmaceutical, cosmetic or cosmeceutical formulation or product further comprises an antimicrobial agent, as an additional pharmaceutically active agent.

Microbial infections include any infection caused by a pathogenic microorganism, including, bacterial infection, fungal infection, protozoal infection, viral infection and the like, e.g., molluscum contagiosum (a viral infection of the skin or occasionally of the mucous membranes), fungal nail infections, and cutaneous leishmaniasis.

Topical bodily sites include skin, mucosal tissue, eye, ear, nose, mouth, rectum and vagina.

In some embodiments, there is provided an article (e.g., a medical device such as a bandage or adhesive patch), a formulation, or a product, as described herein, configured for topical application, whereby a condition treatable by such as article, product or formulation is an infection caused by a microorganism.

In some embodiments of the present invention, the article is e.g., a fabric, a bandage, a wipe (e.g., a wet wipe), a pledget, a swab, a suppository, a dressing, a solution, a mousse, a pad, or a patch.

In some exemplary embodiments of the present invention, the article is in the form of paste, cream, lotion, foam, gel, emulsion, an ointment, or soap.

In some embodiments, the personal care formulation of the present invention can be used to treat skin tissue or on damaged or unhealthy skin tissue.

The phrase "damaged or unhealthy skin tissue" as used herein refers to a deviation from healthy functional skin tissue. In the case of skin- a skin that is weaker, less elastic, and is more prone to injury than healthy skin. The structure of unhealthy or damaged skin is inferior to that of healthy skin (for example, the dermis and epidermis contain fewer cells and collagen).

The phrase "healthy skin tissue" as used herein refers to skin that is strong, elastic, smooth and plump. One purpose of treating healthy skin is to prevent deterioration of skin induced by aging or environmental stress including, but not limited to, microbial infection.

The term "damaged" as used herein, or any grammatical derivative thereof, refers broadly to injuries to the skin and subcutaneous tissue as well as internal organs initiated in any one of a variety of ways (e.g., pressure sores from extended bed rest, wounds induced by trauma, wounds received during or following a surgical procedure and the like) and with varying characteristics. Examples include, but are not limited to, bruises, scrapes, burn wounds, sunburn wounds, incisional wounds, excisional wounds, surgical wounds, necrotizing fascitis, ulcers, venous stasis ulcers, diabetic ulcers, decubitus ulcers, aphthous ulcers, pressure ulcers, scars, alopecia areata, dermatitis, allergic contact dermatitis, atopic dermatitis, berloque dermatitis, diaper dermatitis, dyshidrotic dermatitis, psoriasis, eczema, erythema, warts, anal warts, angioma, cherry angioma, athlete's foot, atypical moles, basal cell carcinoma, Bateman's purpura, bullous pemphigoid, Candida, chondrodermatitis helicis, Clark's nevus, cold sores, condylomata, cysts, Darier's disease, dermatofibroma, Discoid Lupus Erythematosus, nummular eczema, atopic eczema, dyshidrotic eczema, hand eczema, Multiforme Erythema Nodosum, Fordyce's Condition, Folliculitis Keloidalis Nuchae, Folliculitis, Granuloma Annulare, Grover's Disease, heat rash, herpes simplex, herpes zoster (shingles), Hidradenitis Suppurativa, Hives, Hyperhidrosis, Ichthyosis, Impetigo, Keratosis Pilaris, Keloids, Keratoacanthoma, Lichen Planus, Lichen Planus Like Keratosis, Lichen Simplex Chronicus, Lichen Sclerosus, Lymphomatoid Papulosis, Lupus of the Skin, Lyme Disease, Lichen Striatus, Myxoid Cysts, Mycosis Fungoides, Molluscum Contagiosum, Moles, Nail Fungus, Necrobiosis Lipoidica Diabeticorum, Nummular Dermatitis, Onychoschizia, Onychomycosis, Pityriasis Lichenoides, Pityriasis Rosea, Pityriasis Rubra Pilaris, Plantar Warts, Poison Ivy, Poison Oak, Pompholyx, Pseudofolliculitis Barbae, Pruritus Ani and Pityriasis Alba.

Antimicrobial Activity:

According to an aspect of some embodiments of the present invention there is provided a method of inhibiting or reducing or retarding the formation of load of a microorganism and/or the formation of a biofilm, in and/or on an article. The method comprises incorporating in and/or on the article any one of the formulations disclosed herein, including any of the respective embodiments thereof. The article can be any one of the articles described herein.

Such articles take advantage of the improved antimicrobial activity exhibited by the formulations as described herein.

As shown in the Example section, the present inventors have further demonstrated that thymol, linalool, phenoxyethanol and phenyl ethyl alcohol can be used effectively in antimicrobial compositions, and when used in various combinations with each other at define weight ratio (e.g., from about 90: 10 to about 70:30 thymol/linalool, phenoxyethanol, or phenyl ethyl alcohol), these compounds show synergistic activity effect.

In some embodiments, the term synergism, or any grammatical derivative thereof, is defined as the simultaneous action of two or more compounds in which the total response of an organism to the combination is greater than the sum of the individual components. Although many combinations of antimicrobial compounds have been studied, a synergistic effect is rarely revealed and the global use of antimicrobial combinations with synergistically enhanced activity is rather limited.

Some embodiments of this aspect of present embodiments are included hereinabove, under

"The Formulation", and under "Articles comprising the Formulation" and form an integral part of embodiments relating to "Antimicrobial Activity".

Herein "antimicrobial activity" is referred to as an ability to inhibit (prevent), reduce or retard bacterial growth, fungal growth, biofilm formation or eradicate living bacterial cells, or their spores, or fungal cells or viruses in a suspension or in a moist environment.

Herein, inhibiting or reducing or retarding the formation of load of a microorganism refers to inhibiting, reducing, or retarding growth of microorganisms and/or eradicating a portion or all of an existing population of microorganisms. Thus, formulations described herein can be used both in reducing the formation of microorganisms on or in an article, and in killing microorganisms in or on an article or a living tissue.

The microorganism can be, for example, a unicellular microorganism (prokaryotes, archaea, bacteria, eukaryotes, protists, fungi, algae, molds, yeast, euglena, protozoan, dinoflagellates, apicomplexa, trypanosomes, amoebae and the likes), or a multicellular microorganism.

An article, according to these embodiments, can be also a living tissue, for example, a skin or mucosal tissue, as described herein. In the context of the present embodiments, the formulations, articles and methods described herein may be used to produce cell inhibiting surface, or a microbial cell killing surface, that remains active for extended periods. Such an antimicrobial surface may not need additional treatment with antimicrobial compositions, clean-up treatments to effect decontamination and cosmetic painting, thereby simplifying upkeep of the physical condition and appearance of microbial infestation prone surfaces. It is contemplated that in some embodiments, the formulations of the present invention may be easily applied to susceptible surfaces in advance of and/or during exposure to a microbial organism.

In some embodiments, the microorganism comprises bacterial cells of bacteria such as, for example, Gram-positive and Gram-negative bacteria.

In some embodiments, the Gram-positive bacteria are Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus cereus.

In some embodiments, the Gram-negative bacteria are Escherichia coli, Pseudomonas aeuruginosa, and Burkholderia cepacia.

In some embodiments of the present invention, the microorganism is fungi e.g., Candida albicans and Aspergillus niger.

The term "biofilm", as used herein, refers to an aggregate of living cells which are stuck to each other and/or immobilized onto a surface as colonies. The cells are frequently embedded within a self-secreted matrix of extracellular polymeric substance (EPS), also referred to as "slime", which is a polymeric sticky mixture of nucleic acids, proteins and polysaccharides.

In the context of the present embodiments, the living cells forming a biofilm can be cells of a unicellular microorganism (prokaryotes, archaea, bacteria, eukaryotes, protists, fungi, algae, euglena, protozoan, dinoflagellates, apicomplexa, trypanosomes, amoebae and the likes), or cells of multicellular organisms in which case the biofilm can be regarded as a colony of cells (like in the case of the unicellular organisms) or as a lower form of a tissue.

In the context of the present embodiments, the cells are of microorganism origins, and the biofilm is a biofilm of microorganisms, such as bacteria and fungi. The cells of a microorganism growing in a biofilm are physiologically distinct from cells in the "planktonic form" of the same organism, which by contrast, are single-cells that may float or swim in a liquid medium. Biofilms can go through several life-cycle steps which include initial attachment, irreversible attachment, one or more maturation stages, and dispersion. The phrases "antibiofilm formation activity" refers to the capacity of a substance to affect the prevention of formation of a biofilm of bacterial, fungal and/or other cells, and/or to affect a reduction in the rate of buildup of a biofilm of bacterial, fungal and/or other cells, on a surface of a substrate. This activity is also referred to herein as anti-biofouling activity, or antifouling activity.

As used herein, the term "preventing" in the context of antimicrobial, indicates that the growth rate of the microorganism cells is essentially nullified or is reduced by at least 20 %, at least 30 %, at least 40 %, at least 50 %, at least 60 %, at least 70 %, at least 80 %, at least 90 %, including any value therebetween, of the appearance of the microorganism in a comparable situation lacking the presence of formulation or an article containing same. Alternatively, preventing means a reduction to at least 15 %, 10 % or 5 % of the appearance of the microorganism cells in a comparable situation lacking the presence of the formulation or an article containing same.

Methods for determining a level of appearance of a microorganism cells are known in the art. In many products, the anti-odor components attempt to absorb or merely mask the odor. The anti-odor components may act on the underlying bacterial organisms.

In some embodiments of the present invention the linalool masks the thymol's odor, that is, further acts as a smell masking agent. In some embodiments of the present invention liffarome masks the thymol's odor. In some embodiments of the present invention liffarome and capric acid mask the thymol's odor. Smell tests may be used to analyze the effectiveness of a masking agent. For example, a panel of people comprising a significant number and cross-section of people blindly smells the formulation comprising the thymol and/or the masking agent. The results may be statistically analyzed using parametric or non-parametric methods. In order to eliminate the influence of the difference among individual panelists and their physical conditions, in vitro and automated systems are also available to measure smell. Such systems are available from companies such as Odotech Inc.

General:

As used herein the term "about" refers to ± 10 %.

The terms "comprises", "comprising", "includes", "including", "having" and their conjugates mean "including but not limited to".

The term "consisting of" means "including and limited to". The term "consisting essentially of" means that the composition, method or structure may include additional ingredients, steps and/or parts, but only if the additional ingredients, steps and/or parts do not materially alter the basic and novel characteristics of the claimed formulation, method or structure.

The word "exemplary" is used herein to mean "serving as an example, instance or illustration".

Any embodiment described as "exemplary" is not necessarily to be construed as preferred or advantageous over other embodiments and/or to exclude the incorporation of features from other embodiments.

The word "optionally" is used herein to mean "is provided in some embodiments and not provided in other embodiments". Any particular embodiment of the invention may include a plurality of "optional" features unless such features conflict.

As used herein, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" may include a plurality of compounds, including mixtures thereof.

Throughout this application, various embodiments of this invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention.

Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

Whenever a numerical range is indicated herein, it is meant to include any cited numeral (fractional or integral) within the indicated range. The phrases "ranging/ranges between" a first indicate number and a second indicate number and "ranging/ranges from" a first indicate number "to" a second indicate number are used herein interchangeably and are meant to include the first and second indicated numbers and all the fractional and integral numerals therebetween.

As used herein the term "method" refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.

As used herein, the term "treating" includes abrogating, substantially inhibiting, slowing or reversing the progression of a condition, substantially ameliorating clinical or aesthetical symptoms of a condition or substantially preventing the appearance of clinical or aesthetical symptoms of a condition.

It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements.

Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples.

EXAMPLES

Reference is now made to the following examples, which together with the above descriptions illustrate some embodiments of the invention in a non-limiting fashion.

General Procedure:

In exemplary procedure, the minimum inhibitory concentration (MIC) was determined by applying the broth microdilution method and monitoring the bacterial growth using absorbance measurement at 600 nm. The test is performed in ELISA microplates for bacterial strains and Candida albicans. Microbial growth was estimated by reading O.D at 600 nm.

In another exemplary procedure, MIC evaluation test with A. niger was performed in test tubes.

Growth in this case is visually evaluated. A pure culture of a single microorganism was grown in Tryptic soy broth, or sabouraud broth for yeast and mold.

The culture is standardized using standard microbiological techniques to have a concentration of very near 1 million cells per milliliter.

The antimicrobial agent is diluted a number of times, with Tryptic soy broth or sabouraud broth for fungi. After the antimicrobial agent had been diluted, a volume of the standardized inoculums was added to each dilution vessel, bringing the microbial concentration to approximately 500,000 cells per milliliter.

The inoculated, serially diluted antimicrobial agent was incubated at an appropriate temperature for the test organism, 24 hours for bacteria 48 for yeast and 72 for mold.

After incubation, the series of dilution vessels was observed for microbial growth, usually indicated by turbidity and/or a pellet of microorganisms in the bottom of the vessel. The last tube in the dilution series that does not demonstrate growth corresponds with the minimum inhibitory concentration (MIC) of the antimicrobial agent.

For bacterial strains and Candida albicans microbial growth was estimated by reading O.D at

600 nm.

In another exemplary procedure, the antimicrobial preservative effectiveness was tested to measure the efficiency of a preservative. The method applied is based on the USP 30 guidelines for antimicrobial effectiveness testing. The microorganisms used for the test were E. coli ATCC 8739, P.aeruginosa ATCC 9027 , S. aureus ATCC 6538, C. albicans ATCC 10231, A. niger ATCC 16404.

In exemplary procedure, five samples of the product were inoculated with 1 microorganism each. The volume of the culture was calculated to yield a count of 10 ⁵ - 10 ⁶ CFU/gr (CFU: colony forming unit).

In exemplary procedure, a 4 weeks follow-up was performed. Each product was sampled once a week and a viable count was performed. Polysorbate 20 and Soy Lecithin are added to the culture media in order to neutralize the preservatives and enable the recovery of all living microorganisms.

Exemplary formulations are described in Example 1 that follows

EXAMPLE 1

ANTI MICRO-ORGANIC ACTIVITY EVALUATION

Thymol, cinnamaldehyde and Eugenol are known to have antimicrobial activity. They are active compounds of natural extracts. The results below (Table 1) demonstrate that it is possible to mimic a natural extract for preservation.

Table 1 Pseudomonas Staphylococcus Aspergillus Candida

Microorganism/ E. coli

aeruginosa aureus niger albicans Preservative (Gram -)

(Gram -) (Gram +) (Mold) (Yeast)

Cinnamaldehyde 500 500 250 120 62

Thymol 200 800 200 400 200

Eugenol 1200 2250 1200 400 400

From Table 1 it can be concluded that the antimicrobial activity of cinnamaldehyde and thymol are better than the one from eugenol.

It can also be concluded that cinnamaldehyde and thymol have a broad-spectrum activity. However, it is noteworthy that cinnamaldehyde is less chemically stable as it can easily be oxidized, and that thymol has a significant odor therefore smell masking agent was called for.

EXAMPLE 2

SMELL MASKING

Thymol has a significant odor, and therefore smell masking agent was called for. In exemplary procedures, three smell masking agents were examined: vanillin, linalool and menthol. The antimicrobial activity of these agents was tested as shown in Table 2.

Table 2

It can be concluded from Table 2 that linalool exhibits the best antimicrobial activity. Menthol is a crystalline substance with low solubility and does not have significant antimicrobial activity. Vanillin is less chemically stable. Therefore, linalool was chosen as a smell masking agent for the blends due to his antimicrobial activity and pleasant smell in combination with thymol.

EXAMPLE 3

EVALUATION OF THE MINIMAL INHIBITORY CONCENTRATIONS (MIC) Thymol is in the form of a powder and it cannot be easily combined with linalool without a solvent. Therefore, in exemplary procedures, blends with propylene glycol as a solvent were prepared. Different ratios of thymol /linalool were tested in order to find the most effective ratio.

The results are shown in Table 3 hereinbelow.

Table 3

It can be concluded that blends with thymol /linalool ratio 70:30, 80:20, 90: 10 provide the best antimicrobial activity.

However, blends with thymol /linalool ratio 80:20 and 90: 10 have dominant odor of thymol, therefore blend 70:30 (thymol/linalool) was selected to work with. EXAMPLE 4

PRESERVATIVE EFFECTIVENESS (CHALLENGE TESTS)

In another exemplary procedure, the antimicrobial preservative effectiveness was tested to measure the efficiency of a preservative. The sample tested was a face cream.

The Preservation tested was: 1.4% (thymol: 24.5% + linalool: 10.5% + propylene glycol: 65%) pH: 6.6.

Method applied: The method applied is based on the USP 30 guidelines for antimicrobial effectiveness testing. The microorganisms used for the test are listed in the Table 4 below. Five samples of the product are inoculated with one microorganism each. The volume of the culture is calculated to yield a count of 10 ⁵ - 10 ⁶ CFU/gr.

A 4 weeks follow-up was performed. Each product was sampled once a week and a viable count was performed. Polysorbate 20 and Soy Lecithin were added to the culture media in order to neutralize the preservatives and enable the recovery of all living microorganisms. The results are presented in Table 4 below.

Table 4

The test was stopped after a week in view of the high viable counts.The propylene glycol was next replaced by other vehicles as shown below (Example 6). EXAMPLE 5

SYNERGISM BETWEEN THYMOL AND LINALOOL

The synergistic effect of thymol and linalool is presented in Table 5 below.

Table 5 Pseudomonas Staphylococcus Aspergillus Candida

Microorganism/ E. coli

aeruginosa aureus niger albicans Preservative (Gram -)

(Gram -) (Gram +) (Mold) (Yeast)

Thymol=24.5%

Linalool=10.5% 900 2400 600 400 400 Propylene glycol=65%

Amount of Thymol in the

220 588 147 98 98 blend

Thymol as a single

200 800 200 400 200 preservative

Amount of Linalool in the

95 252 60 40 40 blend

Linalool as a single

800 3000-4000 600 500 600 preservative

The data presented in Table 5 show a synergistic action between thymol and linalool.

EXAMPLE 6

ALTERNATIVE VEHICLES

In additional exemplary procedures, three solvents being used in cosmetic industry: benzyl alcohol, phenoxyethanol, phenyl ethyl alcohol, were examined as preservative effectiveness, according to the procedures described hereinabove.

Tables 6 and 7 below present the preservative effectiveness (MIC tests).

Table 6

Pseudomonas Staphylococcu Aspergillus Candida

Microorganism/ E. coli

aeruginosa s aureus niger albicans Preservative (Gram -)

(Gram -) (Gram +) (Mold) (Yeast)

Thymol=24.5%

Linalool=10.5% 800 2400 400 300 400 Benzyl alcohol=65%

Thymol=24.5%

Linalool=10.5%

600 1800 400 300 400 Phenyl ethyl alcohol

=65%

Thymol=24.5%

Linalool=10.5% 800 2000 400 300 400 Phenoxyethanol=65 %

Table 7

It can be concluded that blends with benzyl alcohol, phenyl ethyl alcohol and phenoxyethanol as solvents have better microbiological activity than blends with Propylene glycol. It can also be concluded that blend with phenyl ethyl alcohol has a better activity than blends with phenoxyethanol and benzyl alcohol.

EXAMPLE 7

PRESERVATIVE EFFECTIVENESS (CHALLENGE TESTS) In another exemplary procedure, the antimicrobial preservative effectiveness was tested to measure the efficiency of a preservative. The sample tested was a face cream.

Method: The method applied was based on the USP 30 guidelines for antimicrobial effectiveness testing. The microorganisms used for the test are listed in the table below.

Five samples of the product were inoculated with 1 microorganism each. The volume of the culture was calculated to yield a count of 10 ⁵ - 10 ⁶ CFU/gr.

A 4 weeks follow-up is performed. Each product was sampled once a week and a viable count was performed. Polysorbate 20 and Soy Lecithin were added to the culture media in order to neutralize the preservatives and to enable the recovery of all living microorganisms.

The preservation composition was:

A) Preservation: 1% (Thymol: 24.5% + Linalool: 10.5% + Phenoxyethanol:65%), pH: 6.7

The results are shown in Table 8.

Table 8

In additional exemplary procedures, the sample tested was a face cream and the preservation composition was:

B) Preservation: 1% (Thymol: 24.5% +Linalool: 10.5%+ Benzyl alcohol: 65%), pH: 6.9

The results are shown in Table 9 below.

Table 9

E. coli P.aeruginosa S. aureus C. albicans A. niger

ATCC 8739 ATCC 9027 ATCC 6538 ATCC 10231 ATCC 16404 Inoculum 2.2* 10 ⁵ 4.4* 10 ⁵ 4.3* 10 ⁵ 2.4* 10 ⁵ 2.1* 10 ⁵

7 days <100 <100 3*10 ³ 1*10 ² <100

14 days <10 <10 <10 <10 <10

21 days <10 <10 <10 <10 <10

28 days <10 <10 <10 <10 <10

It can be concluded t lat the tested proc uct meets the USP criteria for cosmetics. In additiona exemplary procedures, the sample tested was a face cream and the preservation composition was:

C) Preservation: 1% (Thymol: 24.5% + Linalool: 10.5%+ Phenyl ethyl alcohol :65%), pH: 6.9. The results are shown in Table 10 below.

Table 10

It can be concluded that the tested product meets the USP criteria for cosmetics.

EXAMPLE 8

ADDITIONAL SMELL MASKING

As previously stated, Thymol has a significant odor, and therefore requires a smell masking agent. Fifteen smell masking agents were tested to enhance the odor of the thymol, linalool and phenoxyethanol composition. Liffarome, 4-carvomenthenol, carvacrol, benzoic acid, sorbic acid, citric acid, capric acid, lactic acid, palmitic acid, sebacic acid, myristic acid, undecylenic acid, citronellic acid, linalyl acetate, and alpha-bisabolol were all found to be ineffective in masking the odor that was still present in the Thymol: 24.5% + Linalool: 10.5% + Phenoxyethanol: 65% composition (Table 11, ++ :good masking, + :low masking, 0 :poor masking, - :no masking). Table 11

Substrate blend Name of the olfactive mask Qualitative evaluation of the masking performance

Thymol

Linalool Liffarome + Phenoxyethanol

Thymol

Linalool Liffarome + Capric acid + Phenoxyethanol

Thymol

Linalool 4-Carvomenthenol - Phenoxyethanol

Thymol

Linalool Carvacrol - Phenoxyethanol

Thymol

Linalool Benzoic acid - Phenoxyethanol

Thymol

Linalool Sorbic acid - Phenoxyethanol

Thymol

Linalool Citric acid - Phenoxyethanol

Thymol

Linalool Capric acid 0 Phenoxyethanol

Thymol

Linalool Lactic acid - Phenoxyethanol

Thymol

Linalool Palmitic acid - Phenoxyethanol

Thymol

Linalool Sebacic acid - Phenoxyethanol

Thymol

Linalool Myristic acid - Phenoxyethanol

Thymol

Linalool Undecylenic acid - Phenoxyethanol

Thymol

Linalool Citronellic acid 0 Phenoxyethanol Thymol

Linalool Linalyl acetate - Phenoxyethanol

Thymol

Linalool alpha-Bisabolol - Phenoxyethanol

Thymol

Liffarome ++

Phenoxyethanol

Thymol

Capric acid +

Phenoxyethanol

Thymol

Liffarome + Capric acid ++

Phenoxyethanol

Thymol

4-Carvomenthenol - Phenoxyethanol

However, upon removal of linalool from the composition it was found that both liffarome and liffarome with capric acid were fully capable of masking the thymol odor (Table 11). Further, as phenoxyethanol is also an effective antimicrobial agent, removal of linalool did not reduce the efficacy of the compound (Table 12). Additionally, the presence of two antimicrobial agents still decreases the chance of development of resistance to the composition.

Table 12

Microorganism/ Staphylococcus Pseudomonas Aspergillus Candida

E. coli

Preservative aureus aeruginosa niger albicans

(Gram -)

(Gram +) (Gram -) (Mold) (Yeast)

Thymol= 24.5%

Phenoxyethanol=

75.38% 400 800 2000 300 400 Liffarome=

0.12%

Thymol= 24.5%

Phenoxyethanol=

73.88%

Capric acid= 400 800 1800 300 400

1.5%

Liffarome=

0.12% Thymol= 24.5%

Phenyl ethyl

alcohol= 75.38% 400 800 1800 300 400

Liffarome=

0.12%

A similar resu t was found for the Thymol: 24.5% + Linalool: 10.5%+ Phenyl ethyl alcoho

:65% composition. Capric acid, Hffarome, and valine were all unable to improve the thymol odor. By contrast, Hffarome was successful in masking the thymol odor of a composition that lacked linalool (Table 13, ++ :good masking, + :low masking, 0 :poor masking, - :no masking), and did not reduce the antimicrobial efficacy of the compound (Table 12).

Table 13

Liffarome was effective as only 0.12% of the composition by weight, and capric acid was effective as only 1.5% by weight. Thus, the three new blends had the following compositions: Thymol: 24.5% + Phenoxyethanol:75.38% + liffarome: 0.12%; Thymol: 24.5% + Phenoxyethanol:73.88% + liffarome: 0.12% + capric acid 1.5%; and Thymol: 24.5% + Phenyl ethyl alcohol :75.38% + liffarome: 0.12%.

EXAMPLE 9

PRESERVATIVE EFFECTIVENESS (CHALLENGE TESTS)

In another exemplary procedure, the antimicrobial preservative effectiveness was tested to measure the efficiency of a preservative. The sample tested was a face cream. Method: The method applied was based on the USP 35 guidelines for antimicrobial effectiveness testing. The microorganisms used for the test are listed in the table below. Five samples of the product were inoculated with 1 microorganism each. The volume of the culture was calculated to yield a count of 10 ⁵ - 10 ⁶ CFU/gr. A 4 weeks follow-up is performed. Each product was sampled once a week and a viable count was performed. Polysorbate 20 and Soy Lecithin were added to the culture media in order to neutralize the preservatives and to enable the recovery of all living microorganisms .

The preservation composition was:

D) Preservation: 0.8% (Thymol: 24.5% + Phenoxyethanol:75.38% + Liffarome:0.12%), pH: 6.7. The results are shown in Table 14.

Table 14

It can be concluded that the tested product meets the USP criteria for cosmetics. In additional exemplary procedures, the sample tested was a face cream and the preservation composition was:

E) Preservation: 0.8% (Thymol: 24.5% + Phenoxyethanol: 73.88% + Liffarome: 0.12% + Capric acid: 1.5%), pH: 6.9

The results are shown in Table 15.

Table 15

E. coli P.aeruginosa S. aureus C. albicans A. niger ATCC 8739 ATCC 9027 ATCC 6538 ATCC 10231 ATCC 16404

Inoculum 5.5*10 ⁵ 6.3*10 ⁵ 8.2*10 ⁵ 3.1*10 ⁵ 1.8*10 ⁵

7 days <100 <100 1.8*10 ³ <100 2.5*10 ²

14 days <10 <10 <10 <10 20 21 days <10 <10 <10 <10 <10

28 days <10 <10 <10 <10 <10

It can be concluded that the tested product meets the USP criteria for cosmetics.

Although the invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims.

All publications, patents and patent applications mentioned in this specification are herein incorporated in their entirety by reference into the specification, to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent that section headings are used, they should not be construed as necessarily limiting.