A hygiene composition

Abstract

The present invention relates to a cleaning composition that comprises a bacteriophage that delivers long lasting hygiene. This is achieved through inclusion of a specific polysaccharide in the composition that helps the bactericide adhere to the surface which is to be cleaned in the home like those in the bathroom, toilet, and kitchen sink.

Description

[0001] P0001058 CPL

[0002] 1

[0003] A HYGIENE COMPOSITION

[0004] Field of the Invention

[0005] The present invention relates to a composition that comprises a bacteriophage which is included

[0006] 5 for delivering hygiene benefits. Particularly, the present invention relates to delivering long lasting hygiene where the bacteriophage adheres on to the surfaces being cleaned in the home like those in the bathroom, toilet, and kitchen sink.

[0007] Background of the Invention

[0008] People like the surfaces of homes to be kept clean and free of dust and soil. Additionally, they also like to have cleanliness in other common spaces like offices, restaurants, hotels and public places like bus and train stations and airports. Firstly, consumers like to have a good visual appearance of cleanliness and apparent hygienic conditions in addition to the place smelling fresh and pleasant. Further, there are places in the bathroom, toilet and kitchen sinks which are vertical

[0009] 15 or inclined which are difficult to clean and maintain hygiene. This is because the cleaning composition slides off the vertical / inclined surfaces such that the antimicrobial active included therein reside on the surface for a very short time. Thus, delivering long lasting hygiene by way of ensuring adherence of the active on to the surface is a problem. This is especially true when the composition is of the wash-off type and comprises high amount of surfactant which are

[0010] 20 generally rinsed off from the surfaces after application of the composition. A very effective and targeted method of delivering anti-bacterial efficacy is through use of bacteriophages.

[0011] Bacteriophages (also called phages) are bacterial viruses that attach only to their specific host bacteria and kill them by bacterial lysis. Phages are thus very specific in that they only attack their targeted bacterial hosts. They cannot infect humans or other eukaryotic cells.

[0012] The present inventors in trying to ensure adherence of bacteriophages on to the desired surfaces so as to ensure long lasting hygiene found to their surprise that inclusion of a specific polysaccharide viz. pectin in cleaning composition comprising bacteriophages, they were able to get the desired benefit.

[0013] 30

[0014] The use of such natural and ecofriendly actives provides for control of undesirable bacteria for a long period of time, thus offering a good sanitation method that is sustainable, while offering a solution which has minimal side effects usually caused by usage of chemical actives. P0001058 CPL

[0015] 2

[0016] It is therefore an object of the present invention to provide for a composition for enhancing adhesion of bacteriophages on to surfaces.

[0017] Summary of the Invention

[0018] 5 The first aspect of the present invention relates to a hygiene composition comprising

[0019] (i) a bacteriophage capable of lysing bacteria prevalent in homes;

[0020] (ii) pectin;

[0021] (iii) 0.1 to 30 wt% surfactant; and

[0022] (iv) 50 to 98 wt% water.

[0023] Another aspect of the present invention relates to ensuring enhanced adherence of bacteriophage on a surface comprising the step of contacting the surface with a composition of the first aspect, preferably diluted with water.

[0024] Detailed Description of the Invention

[0025] For the avoidance of doubt, any feature of one aspect of the present invention may be utilized in

[0026] 15 any other aspect of the invention. The word "comprising" is intended to mean "including" but not necessarily "consisting of” or "composed of". Thus, the term "comprising" is meant not to be limiting to any subsequently stated elements, but rather to optionally also encompass nonspecified elements of major or minor functional importance. In other words, the listed steps or options need not be exhaustive. Whenever the words "including" or "having" are used, these

[0027] 20 terms are meant to be equivalent to "comprising" as defined above. It is noted that the examples given in the description below are intended to clarify the invention and are not intended to limit the invention to those examples per se. Except in the examples, or where otherwise explicitly indicated, all numbers in this description indicating amounts of material or conditions of reaction, physical properties of materials and / or use are to be understood as modified by the word "about". Unless specified otherwise, numerical ranges expressed in the format "x to y" are understood to include x and y. When for a specific feature multiple preferred ranges are described in the format "x to y", it is understood that all ranges combining the different endpoints are also contemplated. Unless specified otherwise, amounts as used herein are expressed in percentage by weight based on total weight of the composition and is abbreviated as “wt%”.

[0028] 30

[0029] The composition as per the present invention comprises a bacteriophage capable of lysing bacteria prevalent in homes. By “home” is meant any indoor environment that includes houses inhabited by people and other structures like hotels, restaurants, offices, malls, train stations, airports, bus station and other buildings that are frequented by humans and which needs to be P0001058 CPL kept clean and hygienic. The bacteria is preferably selected from one or more of E.coli, Pseudomonas aeruginosa, Enterococcus hirae, Staphylococcus aureus, Morganella morganii, Citrobacter koseri, Proteus vulgaris, Proteus mirabilis, Streptococcus agalactiae, Staphylococcus arlettae, Enterococcus faecalis, Staphylococcus saprophyticus, Bacillus safensis,

[0030] 5 Staphylococcus hominis or Staphylococcus gallinarum. More preferably the bacteria is chosen from one or more of Morganella morganii, Citrobacter koseri, or Proteus vulgaris.

[0031] Preferably, the composition comprises bacteriophage capable of lysing bacteria selected from one or more of E. coli, Pseudomonas aeruginosa, Staphylococcus aureus, Citrobacter koseri, Morganella morganii, and Proteus vulgaris. More preferably, the composition comprises

[0032] 10 bacteriophage capable of lysing bacteria E.coli and a bacteriophage capable of lysing bacteria Pseudomonas aeruginosa. The bacteriophage is preferably included in an amount in the range of 102to 1011PFU / rnl of the composition. It is preferably included in 0.001 to 1%, more preferably 0.01 to 0.5% by weight of the composition.

[0033] The composition of the invention comprises pectin. Pectin is a common polysaccharide found in many plants in nature. Various sources of pectin are available, including apples, citrus peel, gooseberries, quince, pears and plums. Pectin for use in the present invention may be from any of the above sources, preferably from citrus peel, apple or sugar beet, or mixtures thereof. Pectin consists predominantly of • -D galacturonic acid units, but also contains some amount of neutral

[0034] 20 sugars such as rhamnose, xylose, arabinose, galactose and glucose.

[0035] The percentage of galacturonic acid units is therefore the amount of galacturonic acid groups present relative to the total amount of the pectin. The galacturonic acid content of a sample can be determined by methods known in the art, such as for example the Saeman hydrolysis method

[0036] 25 (Englyst and Cummings (Analyst, 109(7), 937-942 (1984), Filisetti-Cozzi and Carpita (Analytical Biochemistry, 197, 157-162 (1991 )).

[0037] Galacturonic acid typically has the following repeat unit structure: P0001058 CPL

[0038] 4

[0039] It is possible to esterify galacturonic acids in the carboxylic acid group. The percentage of esterified units is called the degree of esterification (DE). The degree of esterification can be determined according to methods known in the art, such as the base titration method (Shultz, 1965) as proposed by the Food Chemical Codex (FCC (1981 ). 3rd ed., (1981 ) National Academy

[0040] 5 of Science, Washington, DC), quantification of methanol released during de-esterification using gas chromatography (GC) (Walter et al. (1983), Journal of Food Science, 48: 1006-10070), colorimetry (Hou et al. (1999), Botanical Bulletin of Academia Sincia, 40:1 15-119), high performance liquid chromatography (HPLC) (Levigne S., et al. (2002), Food Hydrocolloids 16: 547-550), nuclear magnetic resonance (NMR) (Rosenbohm et al. (2003) Carbohydrate Research, 338: 637-649) and capillary zone electrophoresis (CZE) (Williams et al. (2003), Journal of Agricultural Food and Chemistry, 51 : 1777-1781 ).

[0041] A method to separate pectin into fractions with different DE is described, for example, by Strom, et al. (2005), Carbohydrate Polymers, Volume 60, Issue 4, 20 June 2005, Pages 467-473.

[0042] 15

[0043] In nature, the carboxylic groups are typically methylated to varying degrees to provide pectin methyl esters. The number fraction of carboxylic acid groups methylated is known as the degree of methyl esterification.

[0044] 20 The pectin used in the composition of the present invention preferably has a degree of methyl esterification higher than 50 wt%, more preferably 52.5 wt% or higher, furthermore preferably 55 wt% or higher, yet more preferably 57.5 wt% or higher and yet furthermore preferably 60 wt% or higher, and most preferably 62.5wt% or higher.

[0045] The pectin used according to the invention preferably has a molecular weight of higher than 50kDa, preferably higher than 100kDa, more preferably higher than 150 kDa. Typical upper limits of pectin molecular weights are of the order of 1000kDa, preferably 800kDa. Thus, the pectin for use in the composition of the invention has a molecular weight of between 50 kDa and 1000kDa.

[0046] 30 The molecular weight of the pectin used in the present invention is determined using Triple Detection Size Exclusion Chromatography (TD-SEC)

[0047] The analysis is conducted using the Malvern OMNISEC system. The complete OMNISEC system consists of OMNISEC RESOLVE (chromatography module) and OMNISEC REVEAL (detectors module). Using multi-detector technologies, it combines refractive index, absorbance, light P0001058 CPL

[0048] 5 scattering and viscometer detectors to measure concentration, molecular weight and other parameters. The data are analyzed using the OMNISEC software.

[0049] Depending on the molecular weight of the polysaccharide, appropriate aqueous columns such as the A6000 and A7000 set of columns and the corresponding guard columns are used. Depending

[0050] 5 on the nature of the polysaccharide to be analyzed (whether they have anionic, neutral or cationic functional groups), routinely used eluents of choice are demineralized water containing 0.02wt% Na azide as the preservative or buffer solutions such as Na or Li nitrate (0.05-0.1 M), buffered to the appropriate pH range using NaOH or acetic acid. Polysaccharides with hydrophobic functionality can be analyzed using 10-20% MeOH.

[0051] Depending on the molecular weight range of the polysaccharides, typical sample solutions are prepared at 0.5 mg / ml and then filtered using 0.45* m Nylon filters prior to dispensing into sample vials. The eluent is filtered using a 0.2 • m Nylon filter. The flow rate throughout all runs is typically at 1 ml / min; the detector and column oven temperatures are set to 40°C.

[0052] 15 The detector calibration is carried out using a narrow molecular weight standard to calculate the detector offsets, the detector constants for all detectors and the band broadening and tailing corrections. The calibration is performed using a standard of known concentration, molecular weight, dispersity and intrinsic viscosity; in this case the narrow molecular weight standard of choice usually is a Pullulan standard material.

[0053] 20

[0054] A verification standard is also run with every sequence. This standard is usually a broad (polydisperse) standard, whose properties are known and can be measured. This independently verifies that the calculation method has been correctly calibrated. For polysaccharide analyses, a Dextran 73KDa material is chosen.

[0055] 25 For accuracy (and to allow for the possibility of sample impurities being present), a fixed dn / dc method is used to calculate the molecular weight data for all samples assuming a dn / dc of 0.147. Pectin is preferably included at a concentration of 0.01 to 20%, preferably 0.1 to 10% by weight of the composition.

[0056] The composition of the invention comprises a surfactant, in 0.1 to 30 wt%. The surfactant that is used depends on the type of surface being cleaned. The surfactant may be one or more of anionic, non-ionic, amphoteric or zwitterionic type. It has been observed that the most preferred surfactant for use in the present invention, independent of the type of surface or type of soil to be cleaned, is an anionic or non-ionic surfactant, preferably a non-ionic surfactant. A non-ionic

[0057] 35 surfactant of the alcohol ethoxylate is especially preferred. Preferably, the composition comprises P0001058 CPL

[0058] 6 from 1 to 10 wt% non-ionic surfactant based on the total weight of composition. A preferred class of nonionic surfactant for use in the invention includes aliphatic Cs to C , more preferably C12 to C15 primary linear alcohol ethoxylates with an average of from 3 to 20, more preferably from 5 to 10 moles of ethylene oxide per mole of alcohol.

[0059] 5

[0060] The composition of the invention preferably comprises a hydrotrope. Suitable hydrotropes for inclusion in the composition of the invention are one or more of an alkali metal cumene sulphonate, alkali metal xylene sulphonates, polyethylene glycol (PEG), mono propylene glycol (MPG), and dipropylene glycol (DPG). The preferred alkali metal cumene sulphonate is sodium cumene sulphone (SCS). The preferred alkali metal xylene sulphonate is sodium xylene sulphonate (SXS). The most preferred hydrotrope for use in the present invention is one or both of SCS and MPG. The composition preferably comprises 0.05% to 5 wt%, more preferably 0.1 to 4 wt% hydrotrope.

[0061] 15 It is especially preferred that the composition of the invention includes a preservative such as isothiazolin derivatives: 5-chloro-2-methyl-4-isothiazolin-3-one (CIT), Benzisothiazolinone (BIT), Methylisothiazolinone (MIT), or Octylisothiazolinone (OIT); alkane diols like 1 ,2 propanediol, 1 ,3 propanediol; lactic acid or sodium benzoate. It is further preferred that the composition of the invention includes a fragrance. Fragrance is also referred to as a fragrance component, a perfume or a perfume component and so these words / phrases are used interchangeably. Perfume components may comprise odoriferous materials and / or profragrance materials. The cleaning composition preferably comprises 0.1 to 15 wt.% perfume, more preferably 0.5 to 8 wt. % free perfume.

[0062] 25 Particularly preferred perfume components are blooming perfume components and substantive perfume components. Blooming perfume components are defined by a boiling point less than 250°C and a Log P or greater than 2.5. Substantive perfume components are defined by a boiling point greater than 250°C and a Log P greater than 2.5. Boiling point is measured at standard pressure (760 mm Hg). Preferably a perfume composition will comprise a mixture of blooming

[0063] 30 and substantive perfume components. The perfume composition may comprise other perfume components.

[0064] The composition of the invention is preferably in liquid form. When in liquid form it comprises 50 to 98 wt% water, preferably from 85 to 95 wt% water. Composition preferably has a pH in the P0001058 CPL

[0065] 7 range of 4.0 to 9.0, preferably from 4.0 to 5.5. The pH of such a liquid composition is measured at 25 °C using a pH meter.

[0066] The composition of the invention may be delivered as products which are specifically with the following application in mind.

[0067] 5

[0068] General purpose and toilet cleaning composition:

[0069] The composition of the invention may be formulated as a general-purpose cleaner which is usually used to clean floors (in which case it may be known as a floor cleaner) or used to clean toilets (in which case it may be known as a toilet cleaner). It may also be used to clean other hard surfaces like furniture, tabletops, kitchen platforms and other surfaces in homes, offices, restaurants and other public places.

[0070] Ingredients like amphoteric surfactant and sequestrant may be included in the floor cleaner and toilet cleaner composition of the present invention.

[0071] General purpose cleaners and toilet cleaners may be diluted before use. When diluted, they may

[0072] 15 be diluted with water in a weight ratio in the range of 1 :10 to 1 :1000.

[0073] Kitchen cleaning composition:

[0074] The composition of the invention is useful for upkeep of areas in the kitchen e.g the sink, cleaning implement like sponge and scrub which may be cleaned using the dishwash composition,

[0075] 20 preferably in liquid form. Various ingredients other than the essential ingredients claimed in the present invention may be used as summarized below.

[0076] When formulated as a dishwashing composition, it may additionally include an anionic surfactant. Further preferably, the composition may comprise an amphoteric surfactant. Preferably, the amphoteric surfactants which may be used are the same as the ones listed under General purpose and toilet cleaning compositions. Amphoteric surfactant, when included, may be present in an amount ranging preferably from 0.1 to 5 wt%, more preferably from 0.1 to 4 wt%, even more preferably from 1 to 3 wt%.

[0077] Preferably, the composition may be used as is, i.e. neat, or it may be diluted before use. The extent of dilution is generally dependent on market choice. In some markets a more concentrated

[0078] 30 product is desired while in others a more diluted product is preferred. When the composition is liquid dishwash, it is typically diluted with water in a weight ratio between 1 :1 to 1 :10.

[0079] Another aspect of the present invention relates to a method of ensuring enhanced adherence of bacteriophage on a surface comprising the step of contacting the surface with a composition of P0001058 CPL the first aspect preferably diluted with water. The composition may be diluted with water in a ratio of composition: water in the range of 1 : 1 to 1 :500, preferably in the range of 1 :1 to 1 :100, more preferably in the range of 1 :2 to 1 :25 and most preferably in the range of 1 :5 to 1 :10. The composition thus contacted on the surface may thereafter be rinsed to be substantially free the

[0080] 5 composition. Such a method ensures prolonged hygiene benefits to a surface including cleaning and stain removal.

[0081] The invention will now be illustrated with the help of the following non-limiting examples.

[0082] Examples

[0083] Examples A,B, 1 -3: Effect of inclusion of pectin in a general-purpose cleaning composition comprising bacteriophage

[0084] A composition used for general purpose cleaning of floor and toilets in homes as shown in Table -1 below was taken:

[0085] Table -1 :

[0086] In the above table

[0087] Bacteriophage used was M13, capable of lysing bacteria E coli (ATCC 15597)

[0088] The above composition was taken, and different concentrations of pectin were added to it as shown in Table -2 below and the adherence of bacteriophage to ceramic tiles was measured

[0089] 20 using the procedure as given below: P0001058 CPL

[0090] 9

[0091] A clean tile was dipped in the cleaning composition with different concentration of pectin (0.1 to 0.5 %) with 104PFU / ml bacteriophage targeting E coli. The samples were incubated for 24 hrs at 37°C. A gentle water wash was given to the tile after incubation. A swab was taken from the surface of the tile, serially diluted and plated in LB agar. The plates were incubated for 24 hrs at

[0092] 5 37°C. Plaques were counted and the number of bacteriophages was enumerated.

[0093] The data on adherence is summarized in Table -2 below:

[0094] Table -2

[0095] 10 Pectin used was GENU Pectin 105 Rapid set from CP Kelco, UK. The sample used had a degree of methyl esterification of 60%.

[0096] The data in Table - 2 above indicates that pectin is highly effective in helping bacteriophage adhere to surfaces when they are cleaned with a surfactant containing cleaning composition.

Claims

P0001058 CPLClaims1. A hygiene composition comprising(i) a bacteriophage capable of lysing bacteria prevalent in homes;(ii) pectin;(iii) 0.1 to 30 wt% surfactant; and(iv) 50 to 98 wt% water.

2. A composition as claimed in claim 1 comprising bacteriophages capable of lysing bacteria chosen from one or more of E.coli, Pseudomonas aeruginosa, Enterococcus hirae, Staphylococcus aureus, Morganella morganii, Citrobacter koseri, Proteus vulgaris, Proteus mirabilis, Streptococcus agalactiae, Staphylococcus arlettae, Enterococcus faecalis, Staphylococcus saprophyticus, Bacillus safensis, Staphylococcus hominis or Staphylococcus gallinarum.

3. A composition as claimed in claim 1 comprising bacteriophages capable of lysing bacteria chosen from one or more of E. coli, Pseudomonas aeruginosa, Staphylococcus aureus, Citrobacter koseri, Morganella morganii, and Proteus vulgaris.

4. A composition as claimed in any one of the preceding claims comprising 0.001 to 1% bacteriophage by weight of the composition.

5. A composition as claimed in any one of the preceding claims comprising 0.01 to 20 wt% pectin.

6. A composition as claimed in any one of the preceding claims wherein the surfactant is anionic or non-ionic.

7. A composition as claimed in claim 6 wherein the surfactant is non-ionic.

8. A composition as claimed in any one of the preceding claims comprising 1 to 10 wt% surfactant.

9. A composition as claimed in any one of the preceding claims in liquid form.P0001058 CPL1110. A composition as claimed in any one of the preceding claims having a pH in the range of 4.0 to 10.0.

11. A method of ensuring enhanced adherence of bacteriophage on a surface comprising the step of contacting the surface with a composition as claimed in any one of the preceding claims, preferably diluted with water.

12. A method as claimed in claim 11 wherein the composition ensures prolonged hygiene benefits to a surface including cleaning and stain removal.

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