COMPOSIÇÃO PARA REDUÇÃO DE MAU ODOR PROVENIENTE DE URINA, MÉTODO DE REDUÇÃO DE MAU ODOR PROVENIENTE DE URINA DE UMA SUPERFÍCIE POR ATÉ 48 HORAS APÓS A LIMPEZA DA SUPERFÍCIE E MÉTODO DE INIBIÇÃO DE FORMAÇÃO DE CALCÁRIO SOBRE UMA SUPERFÍCIE

Abstract

The present invention relates to a composition that is used to reduce malodour originating from urine. Particularly, the present invention relates to a composition that reduces urine malodour by specifically targeting bacteria that have been identified as causing the malodour. This has been achieved by combining certain polysaccharides with bacteriophages that lyse the malodour causing bacteria.

Description

1 / 12 COMPOSITION FOR REDUCING BAD ODOR FROM URINE, METHOD FOR REDUCING BAD ODOR FROM URINE ON A SURFACE FOR UP TO 48 HOURS AFTER CLEANING THE SURFACE, AND METHOD FOR INHIBITING LIMESCALE FORMATION ON A SURFACE. Field of Invention

[0001] The present invention relates to a composition that is used to reduce the bad odor emanating from urine. In particular, the present invention relates to a composition that reduces the bad odor emanating from urine by acting specifically on the bacteria that have been identified as causing the bad odor. Background of the Invention

[0002] People like surfaces in their homes to be kept clean and free of dust and dirt. Furthermore, they also appreciate cleanliness in other common spaces such as offices, restaurants, hotels, and public spaces like bus and train stations and airports. Beyond the visual appearance of cleanliness and apparent hygiene, they also like to have a pleasant odor when occupying such places. In addition to common areas, restrooms and bathrooms are areas where a higher degree of cleanliness and deodorization is desired.

[0003] Cleaning and deodorizing could be achieved by using high-level surfactants in the cleaning composition or by including bleaching agents such as chlorine or peroxides. These chemicals ensure the solubilization of the dirt responsible for the dirty appearance and the extermination of the germs responsible for generating bad odor.

[0004] Although chemicals such as surfactants and bleaches are effective in providing instant cleaning, they have the following disadvantages. Firstly, these cleaning methods do not provide long-lasting cleaning and deodorization. A few hours after the cleaning action and Petition 870250084278, dated 09 / 18 / 2025, page 7 / 45 2 / 12 Disinfection: Dirt tends to accumulate, followed by the formation of a concomitant bad odor. Secondly, the use of aggressive chemicals sometimes tends to lead to an unpleasant chlorine odor, which is also unpleasant for many people. To mask the odors of aggressive chemicals and neutralize residual bad odors, pleasant-smelling perfumes are added to cleaning compositions, but such perfumes also have limited effectiveness and tend to dissipate quickly. The above problems are further aggravated in areas intended for human excrement and cleaning, such as bathrooms and toilets. These areas are used by people at regular intervals, where, despite the use of copious amounts of water for flushing, a constant level of bad odor from urine remains, which is difficult to mask.

[0005] The present inventors, in seeking to solve the above problem, used their knowledge and experience in the field of targeted hygiene where biochemical or microbiological methods are used to specifically target only the bacteria responsible for generating bad odor, for example, bacteriophages that specifically perform the lysis of identified bad odor-causing bacteria.

[0006] Bacteriophages are bacterial viruses that bind only to their specific host bacteria and exterminate them through bacterial lysis. Phages are therefore very specific in that they bind only to their target bacterial hosts. They cannot infect human cells or other eukaryotic cells.

[0007] Although it may seem obvious to use these bacteriophages, the present inventors have found, to their surprise, that combining these bacteriophages with certain selected polysaccharides resulted in an improved benefit in reducing the generation of this bad odor. The benefit of reducing the bad odor is not only observed immediately after surface treatment, but has also proven effective for a long period (48 to 96 hours) after surface treatment. The composition and method of Petition 870250084278, dated 09 / 18 / 2025, page 8 / 45 3 / 12 of the present invention are therefore seen to provide the benefit of reducing bad odor in the long term.

[0008] Thus, the use of these natural and environmentally friendly active ingredients provides control of undesirable bacteria that can cause serious problems, such as disease and contamination or microbiome imbalance with problems associated with bad odor. The present approach of using bacteriophages and polysaccharides thus offers a good sanitization method that is sustainable and effective in removing specific bacteria, while offering a solution that has minimal side effects generally caused by the use of chemical active ingredients.

[0009] In addition to reducing bad odor, it has also been surprisingly found that the present invention inhibits limescale formation.

[0010] Patent document US2016215273 (Intralytix) discloses isolated bacteriophages exhibiting specificity and lytic activity against strains of Shigella species, methods of using the bacteriophages, progeny and derivatives thereof to control the growth of Shigella species in various scenarios. This publication, while disclosing the use of bacteriophages as a general method, does not teach how this approach can be used for reducing urine odor through interaction with specific polysaccharides.

[0011] Therefore, it is an objective of the present invention to provide a composition to reduce the bad odor emanating from urine.

[0012] Another objective of the present invention is to provide such a reduction in bad odor from urine, which is considered long-lasting, thus guaranteeing the reduction of bad odor for up to 48 hours after treatment.

[0013] It is yet another objective of the present invention to provide a composition that inhibits the formation of limescale. Summary of the Invention

[0014] The first aspect of the present invention relates to a composition for reducing the bad odor emanating from urine, comprising: Petition 870250084278, dated 09 / 18 / 2025, p. 9 / 45 4 / 12 (i) 102 to 1011 PFU / mL of bacteriophages capable of lysing selected bacteria from one or more of M. morganii, C. koseri, P. mirabilis, P. vulgaris and E. coli; and (ii) 0.01 to 20% by weight of a polysaccharide that is a combination of pectin and one or both of dextrin and resistant dextrin.

[0015] Another aspect of the present invention relates to a method of reducing urine odor from a surface for up to 48 hours after cleaning the surface, comprising the step of contacting the surface with a composition of the first aspect, preferably diluted in water.

[0016] Yet another aspect of the present invention relates to a method of inhibiting limescale formation on a surface comprising the step of treating the surface with a composition of the first aspect. Detailed Description of the Invention

[0017] For the avoidance of doubt, any feature of one aspect of the present invention may be used in any other aspect of the invention. The term “comprising” is intended to mean “including,” but not necessarily “consisting of” or “composed of.” Therefore, the term “comprising” is intended to mean not to be limiting to any subsequently stated elements, but rather to also optionally encompass unspecified elements of greater or lesser functional importance. In other words, the listed steps or options need not be exhaustive. Whenever the terms “including” or “having” are used, these terms are intended to be equivalent to “comprising,” as defined above. Note that the examples provided in the description below are intended to clarify the invention and are not intended to limit it to those examples alone.Except in examples, or where explicitly stated otherwise, all numbers in this description indicating quantities of material or reaction conditions, physical properties of materials and / or use should be understood as modified by the term “approximately”. Unless otherwise specified, numerical ranges expressed in the format “xay” are understood to include x and y. Petition 870250084278, dated 09 / 18 / 2025, page 10 / 45 5 / 12 y. When, for a specific characteristic, several preferred ranges are described in the format “xay”, it is understood that all ranges combining the different endpoints are also included. Unless otherwise specified, quantities as used herein are expressed as a percentage by weight based on the total weight of the composition and are abbreviated as “% by weight”.

[0018] The composition according to the present invention comprises a bacteriophage capable of lysing selected bacteria from one or more of M. morganii, C. koseri, P. mirabilis, P. vulgaris, and E. coli. Preferably, the composition comprises a bacteriophage capable of lysing C. koseri bacteria and a bacteriophage capable of lysing P. vulgaris bacteria. More preferably, the composition comprises a bacteriophage capable of lysing C. koseri bacteria, a bacteriophage capable of lysing P. vulgaris bacteria, and a bacteriophage capable of lysing E. coli bacteria. Most preferably, the composition comprises a mixture of bacteriophages capable of lysing M. morganii, C. koseri, P. mirabilis, P. vulgaris, and E. coli bacteria. The bacteriophage is preferably included in an amount in the range of 10² to 10¹¹ PFU / mL of the composition.

[0019] The composition of the invention preferably comprises a polysaccharide, which is a combination of pectin and one or both of dextrin and resistant dextrin.

[0020] Pectin is a structural acidic heteropolysaccharide contained in the primary and intermediate lamellae and cell walls of terrestrial plants. Its main component is galacturonic acid, a sugar acid derived from galactose. It is commercially produced as a white to light brown powder, extracted mainly from citrus fruits, and is used in foods as a gelling agent, particularly in jams and preserves. It is also used in dessert fillings, medicines, candies, as a stabilizer in fruit juices and dairy drinks, and as a source of dietary fiber. Pectin, also known as pectic polysaccharides, is rich in galacturonic acid. Petition 870250084278, dated 09 / 18 / 2025, page 11 / 45 6 / 12 galacturonic acid. Several distinct polysaccharides have been identified and characterized within the pectic group. Homogalacturonans are linear chains of D-galacturonic acid linked to α-(1-4). Substituted galacturonans are characterized by the presence of attaching saccharide residues (such as D-xylose or D-apiose in the respective cases of xylogalacturonane and apiogalacturonan) that branch off from a structure of D-galacturonic acid residues. Rhamnogalacturonan pectins (RG-I) contain a repeating disaccharide structure: 4)-α-D-galacturonic(1,2)-αL-rhamnose-(1). From many of the rhamnose residues, the side chains of various neutral sugars branch off. Neutral sugars are primarily D-galactose, L-arabinose, and D-xylose, with the types and proportions of neutral sugars varying depending on the pectin's origin.Another structural type of pectin is rhamnogalacturonan II (RG-II), which is a highly branched, complex, and less frequent polysaccharide.

[0021] Pectin consists predominantly of α-D galacturonic acid units, but also contains some amount of neutral sugars such as rhamnose, xylose, arabinose, galactose and glucose.

[0022] The percentage of galacturonic acid units is therefore the amount of galacturonic acid groups present in relation to the total amount of 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 (Englyst and Cummings (Analyst, 109(7), 937942 (1984), Filisetti-Cozzi and Carpita (Analytical Biochemistry, 197, 157-162 (1991)).

[0023] Galacturonic acid generally has the following repeating unit structure of formula (I): / OH HO (I)

[0024] It is possible to esterify galacturonic acids at the carboxylic acid group. A Petition 870250084278, dated 09 / 18 / 2025, p. 12 / 45 7 / 12 of 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 of Science, Washington, DC), the 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, 40:115-119), 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).

[0025] A method is described for separating pectin into fractions with different DE, for example, by Strom, et al. (2005), Carbohydrate Polymers, Volume 60, Issue 4, 20 June 2005, Pages 467 to 473.

[0026] In nature, carboxylic groups are generally methylated to varying degrees to provide pectin methyl esters. The numerical fraction of methylated carboxylic acid groups is known as the degree of methyl esterification.

[0027] The pectin used in the composition of the present invention preferably has a degree of methyl esterification greater than 50%, more preferably 52.5% or greater, even more preferably 55% or greater, still more preferably 57.5% or greater, and most preferably 60% or greater.

[0028] The pectin used, according to the invention, preferably has a molecular weight greater than 50 kDa, more preferably greater than 100 kDa. Typical upper limits of pectin molecular weights are preferably on the order of 1000 kDa, more preferably 800 kDa. Therefore, the pectin for use in the composition of the invention preferably has a molecular weight between 50 kDa and 1000 kDa.

[0029] The polysaccharide can also be dextrin. Preferably, it is prepared Petition 870250084278, dated 09 / 18 / 2025, page 13 / 45 8 / 12 by means of partial hydrolysis of corn, potato, arrowroot, tapioca, or wheat starches. Hydrolysis can be thermal or enzymatic. Dextrin is a combination of D-glucose units linked through α(1-4) or α(1-6) glycoside linkages. Dextrin generally has the chemical formula (C6H10O5)n. A preferred dextrin is one with CAS No. 9004-53-9. It generally has a pH in the range of 4 to 5 when a 20% solution is measured at 25 °C. It is usually marketed as a light yellow powder that is odorless. The powder generally has a particle size in the range of 100 to 200 µm. It is soluble in water at 20 °C. The viscosity is low, as a 50% solution in water at 20 °C is measured to have a value in the range of approximately 50 to 200 mPa.s. A preferred supplier of dextrin is Roquette Freres, France, which markets it under the trade name Nutriose® FB 06.

[0030] The polysaccharide may also be resistant dextrin. Resistant dextrin is also known by some as maltodextrin. It is usually prepared by highly controlled partial hydrolysis of corn, potato, barley, or wheat. It typically contains 3 to 20 D-glucose units linked by an α(1-4)glycosidic bond. It usually has the chemical formula C6nH(10n+2)O(5n+1). A preferred resistant dextrin has CAS No. 9050-36-6. It is usually marketed as a whitish powder that is odorless. It is soluble in water. A preferred supplier of resistant dextrin is ADM Bio Science & Technology (Tianjin) Co. Ltd., Tianjin, China, which markets it under the trade name Fbersol® 2.

[0031] Of the polysaccharides, pectin is preferred for inclusion in the composition of the invention. The pectin, when included, is preferably derived from one or more of a fruit chosen from pear, apple, guava, plum, currant, orange or any other citrus fruit, preferably citrus. Preferably it is from apple or citrus fruit. The composition preferably comprises the polysaccharide, which is a combination of pectin and one or both of dextrin and resistant dextrin. The polysaccharide is preferably included at a concentration of 0.01 to 20%, preferably 0.1 to 10% by weight of the Petition 870250084278, dated 09 / 18 / 2025, page 14 / 45 9 / 12 composition.

[0032] The composition of the invention can be formulated as a cleaning composition, which is generally used for cleaning floors (in which case it may be known as a floor cleaner) or used for cleaning toilets (in which case it may be known as a toilet cleaner). It can also be used to clean other hard surfaces such as furniture, tabletops, kitchen countertops and other surfaces in homes, offices, restaurants and other public places. Preferably, the composition of the invention is formulated as a general-purpose cleaning composition or a toilet cleaning composition and comprises an anionic or non-ionic surfactant.

[0033] The surfactant is generally anionic in nature. Preferred anionic surfactants are derived from organic sulfates and sulfonates with alkyl radicals containing from approximately 8 to approximately 22 carbon atoms, the term “alkyl” being used to include the alkyl portion of higher acyl radicals. Examples of such materials include alkyl sulfates, alkyl ether sulfates, alkyl sulfonates, alpha-olefin sulfonates, and mixtures thereof. The alkyl radicals preferably contain from 10 to 18 carbon atoms and may be unsaturated. Alkyl ether sulfates may contain from one to ten units of ethylene oxide or propylene oxide per molecule and preferably contain one to three units of ethylene oxide per molecule. The counterion for anionic surfactants is generally an alkali metal, such as sodium or potassium; or an ammonia counterion such as monoethanolamine (MEA), diethanolamine (DEA) or triethanolamine (TEA).Mixtures of these counterions can also be used. Sodium and potassium are preferred.

[0034] The most preferred surfactants are of the alkylbenzene sulfonate type, particularly linear alkylbenzene sulfonates (LAS) with an alkyl chain length of 10 to 18 carbon atoms. Some alkyl sulfate surfactants (PAS) may be used, such as non-ethoxylated primary and secondary alkyl sulfates with an alkyl chain length of 10 to 18. Petition 870250084278, dated 09 / 18 / 2025, p. 15 / 45 10 / 12

[0035] A preferred class of nonionic surfactant for use in the invention includes Cs to C18 aliphatic, more preferably C12 to C15 primary linear alcoholic ethoxylates with an average of 3 to 20, more preferably 5 to 10 moles of ethylene oxide per mole of alcohol. Preferred nonionic surfactants are C16 / 18 alcohol ethoxylates. The composition of the invention preferably comprises 1 to 30% surfactant based on the total weight of the composition.

[0036] Ingredients such as amphoteric surfactant and sequestering agent may be included in the composition of the present invention. General purpose cleaners and toilet cleaners may be diluted before use. When diluted, they may be diluted in water at a weight ratio of 1:10 to 1:1000.

[0037] The composition of the invention can also be supplied as a toilet block. A toilet block is understood to be a solid composition that is fixed to the rim of the toilet bowl, usually in a plastic cage-type container, so that each time the toilet is flushed, a quantity of water passes over the solid composition, corroding a portion of it before falling into the toilet bowl. The toilet block preferably also includes a perfume to provide a pleasant and persistent odor with each flush of the toilet.

[0038] The composition of the invention can also be used to clean fabrics. By fabrics is meant clothing, especially underwear or those used by babies and children that may develop a bad odor from urine. It also includes sheets, blankets and other fabrics used in homes and hospitals that may develop this bad odor. The composition can be supplied as a laundry detergent composition, or as a fabric softener composition. The composition can also be supplied in a spray format.

[0039] According to another aspect of the present invention, a reduction of bad odor from urine on a surface is provided for up to 48 hours after cleaning the surface, comprising the step of contacting the surface with a composition of the invention diluted preferably in water. Petition 870250084278, dated 09 / 18 / 2025, page 16 / 45 11 / 12

[0040] The present invention also provides a method for inhibiting limescale formation on a surface comprising the step of treating the surface with a composition of the present invention.

[0041] The invention will now be illustrated by means of the following non-limiting examples. Examples

[0042] Examples AC, 1-2: Inhibition of foul odor from the urine by means of the MS2 phage

[0043] The inhibition of foul odor from urine was measured using various active ingredients and their combination, as shown in Table 1, with the % inhibition summarized therein. The protocol used to measure the % inhibition is provided below.

[0044] Real urine was collected in sterile sample cups from five volunteers and pooled. This was diluted in a 1:10 ratio with Artificial Urine. In 100 mL glass bottles / containers, 40 mL of the urine solution were added and a glass slide was inserted into each container. The mouthpiece of the container was fitted with a disposable 0.22 µm filter (Nalgene filter), hermetically sealed with parafilm. The test product was applied to the urine solution at different concentrations. The positive control was maintained without the addition of any product / actives.

[0045] The samples were incubated at 37 °C for 24 / 48 hours to allow odor to develop within the container. After 24 / 48 h, odor measurements were performed using an olfactory test with a positive control as maximum odor (score 5) and no odor as score 0. The slides were dried, labeled, and stored. The pectin for use in the present invention was obtained from CP Kelco, UK. MS2 is a bacteriophage known to inhibit E. coli and was obtained from ATCC, USA. Table 1 Example Assets Average Score % of Inhibition A Control 5.00 0.0 Petition 870250084278, dated 09 / 18 / 2025, page 17 / 45 12 / 12 B MS2 (108 PFU / mL) 3.50 30 C 0.1% pectin 3.42 32 1 MS2 + 0.1% pectin 2.83 43 2 MS2 + 0.2% pectin 2.67 47

[0046] The data in Table 1 above indicate that the compositions according to the invention (Examples 1 and 2) provide vastly improved odor inhibition compared to the individual actives (Examples B and C).

[0047] Examples DF, 3-6: Additional examples in the combination of actives

[0048] Additional experiments were carried out using the combination of one or two polysaccharides with an MS2 bacteriophage, as shown in Table 2 below, with the results summarized therein. Table 2: Example Assets Average Score % of Inhibition A Control 5.00 0.0 C 0.1% Pectin 3.42 32 D MS2 (104 PFU / mL) 3.08 38 E 2% Fbersol 2 3.33 33 F 2% Nutriose FB 06 2.67 46 3 MS2 + 2% Fbersol 2 2.75 45 4 MS2 + 2% Nutriose FB 06 2.50 50 5 MS2 + 0.1% pectin + 2% Fbersol 2 2.58 48 6 MS2 + 0.1% pectin + 2% Nutriose FB 06 2.17 57

[0049] The data in Table 2 above indicate improved odor inhibition through the use of a polysaccharide (Examples 3-4) in combination with a bacteriophage (MS2). Further improvement is obtained when two polysaccharides are used in combination with MS2 (Examples 5-6). Petition 870250084278, dated 09 / 18 / 2025, page 18 / 45

Claims

1 / 2 Claims 1. Composition for reducing bad odor from urine, characterized by comprising: (i) 102 to 1011 PFU / mL of bacteriophages capable of lysing bacteria selected from one or more of M. morganii, C. koseri, P. mirabilis, P. vulgaris and E. coli; and (ii) 0.01 to 20% by weight of a polysaccharide that is a combination of pectin and one or both of dextrin and resistant dextrin.

2. Composition according to claim 1, characterized by comprising bacteriophages capable of lysing the bacteria C. koseri and P. vulgaris.

3. Composition according to claim 1, characterized by comprising bacteriophages capable of lysing the bacteria C. koseri, P. vulgaris and E. coli.

4. Composition according to claim 1, characterized by comprising a mixture of bacteriophages capable of lysing the bacteria M. morganii, C. koseri, P. mirabilis, P. vulgaris and E. coli.

5. Composition, according to any of the preceding claims, characterized in that pectin is derived from one or more of a fruit selected from pear, apple, guava, plum, currant, orange and any other citrus fruit, preferably citrus fruit.

6. A composition, according to any of the preceding claims, characterized in being a general-purpose cleaning composition or a toilet bowl cleaning composition, further comprising an anionic or non-ionic surfactant.

7. Composition, according to any of the preceding claims, characterized by being in liquid or solid form.

8. Method for reducing urine odor from a surface for up to 48 hours after cleaning the surface, characterized by comprising the step of contacting the surface with a composition, as defined in any of the preceding claims, preferably diluted in water.

9. Method for inhibiting limescale formation on a surface, characterized by comprising the step of treating a surface with a composition, as defined in any one of claims 1 to 7. Petition 870250084278, dated 09 / 18 / 2025, page 20 / 45