Antimicrobial compositions for hygiene applications

Abstract

Provided herein is an antimicrobial composition for hygiene applications. The antimicrobial composition comprises an effective concentration of a cathelicidin LL-37 peptide fragment in combination with an effective concentration of non- ionic surfactant. The non-ionic surfactant is an Alkyl-Poly Glucoside (APG) or alcohol ethoxylates or both. The antimicrobial composition further comprises of a chelating agent like disodium EDTA, or malic acid or citric acid or GLDA or PES A. The composition achieves log reduction of at least 3 against Gram negative bacteria Pseudomonas aeruginosa, Escherichia coll and Gram positive bacteria Staphylococcus aureus, Enterococcus hirae.

Description

[0001] ANTIMICROBIAL COMPOSITIONS FOR HYGIENE APPLICATIONS

[0002] Field of Invention

[0003] The present invention relates to antimicrobial compositions for topical application. Particularly, it relates to antimicrobial compositions comprising an antimicrobial peptide with one or more surfactant.

[0004] Background and Prior art

[0005] The well-known and currently used antimicrobial actives such as Triclosan mediate bacterial killing by specific mechanism that leads to selective pressure on the microorganism to develop resistance. In organisms such as Pseudomonas aeruginosa the resistance is plasmid mediated leading to faster spread of resistance. World Health Organization (WHO) classifies the appearance of antibiotic resistance as one of the biggest threats to human health. Indiscriminate use of certain antimicrobial actives would exacerbate the problem. There are few active combinations that are efficacious as well as safe.

[0006] The efficacious antimicrobial actives such as Triclosan are under global regulatory agency’s scanner because of safety concern. Triclosan is associated with endocrine disruption and antimicrobial resistance.

[0007] The long-term systemic exposure to actives like Triclosan is an area of concern. Hence, there is a need to develop efficacious and safer antimicrobial active combinations.

[0008] Since their discovery in the 1980s, antimicrobial peptides (AMPs) have been viewed as one of the important solutions to the impending crisis of antimicrobial resistance. Antimicrobial peptides (AMPs), evolutionary ancient and conservative tools of the innate immune system providing immediate response to the large set of various pathogens. And Antimicrobial peptides (AMPs) produced by humans, animals, plant and bacteria. AMPs differ in amino acid sequence and structure, but predominantly are cationic and they can adopt an amphipathic conformation, thus, they are able to easily interact with the negatively charged components on the surface of bacterial cells and integrate into the lipid bilayer. The principle mechanism of antibacterial action of AMPs is related with their ability to alter membrane permeability and damage its structure. This leads to leakage of vital components, ions, and metabolites. Membrane destabilization additionally affects functioning of membrane-associated protein complexes. Some AMPs also have intracellular targets and interfere metabolic process such as DNA and Protein synthesis. Hence, wide-scale multi targeted action is believed to be one of the reasons for the effectiveness of AMPs toward multidrug-resistant bacterial strains and an obstacle for the development of a high resistance level to such compounds.

[0009] Antimicrobial peptides (AMPs) are naturally occurring host defense molecules with potent antimicrobial activity. The native AMPs are known to undergo proteolytic degradation in vivo and the fragments generated carry out diverse functions. The AMPs are considered as next generation antimicrobial molecules after the microorganisms have developed resistance against antibiotics due to their indiscriminate and rampant use.

[0010] Nagaoka et al., Synergistic actions of antibacterial neutrophil defensins and cathelicidins. Inflamm Res. 2000 Feb;49(2):73-9 discloses the individual and synergistic actions of defensins and cathelicidins in the presence of physiological concentration of NaCl. The antibacterial activities of human cathelicidin (CAP18 / LL-37) and guinea pig cathelicidin (CAPH) were resistant to NaCl. The paper concludes that defensins cannot function as antibacterial molecules by themselves, but can synergistically work with cathelicidins to exert the antibacterial activity in the extracellular milieu by augmenting the membrane permeabilization of target cells.

[0011] CN20181570015 discloses striae gravidarum repairing cream containing epidermal stem cell extractive. The striae gravidarum repairing cream is prepared from the following ingredients in mass percent: 0.005 to 5.0% of hyaluronic acid and antibacterial peptide bonded material, 0.1 to 5.0% of nonionic surface active agent, 0.5 to 9.0% of epidermal stem cell extractive, 0.001 to 5.0% of semen coicis polypeptide, 0.001 to 2.0% of palmitoyl oligopeptide and the balance of sterile water. The antibacterial peptide in a formula of the striae gravidarum repairing cream disclosed by the invention has a sterilizing and disinfecting effect; the carrier hyaluronic acid not only can ensure skin moisture, but also can improve skin breathability; the composition has a wide antimicrobial spectrum and can clean and moisturize the skin; furthermore, the striae gravidarum repairing cream has the advantages of gentle formula, no stimulation and higher stability; the striae gravidarum repairing cream contains a lot of biological active factors beneficial to skin regeneration and repairing and can promote self renewal, regeneration and repairing of the skin; meanwhile, when the striae gravidarum repairing cream is matched with the semen coicis polypeptide to be used, an antioxidant effect of the striae gravidarum repairing cream can be further improved, ageing is delayed; furthermore, a repairing effect of the striae gravidarum repairing cream can be further greatly enhanced.

[0012] CN20141591876 discloses a contact lens care composition, which contains, by mass, 0.005-5.0% of a hyaluronic acid and antibacterial peptide bonding substance, 0.1-5.0% of a non-ionic surfactant, 0.5-9.0% of an inorganic salt, 0.001-5.0% of a buffer, 0.001-2.0% of a chelating agent, and the balance of sterilized water. According to the document, the antibacterial peptide in the formula can provide the disinfection and sterilization effect, and with the hyaluronic acid, the wetting of the contact lens can be ensured, and the gas permeation of the contact lens can be improved; and the contact lens care composition has characteristics of broadspectrum antibacterial property, cleaning property, moisturizing, mild formula, no irritating and high stability, can be used for cleaning, disinfecting and storage of the contact lens, and can further be used as the wetting agent when wearing the contact lenses. US 7,960,339 B2 relates to families of polypeptides and lipopolypeptides that have antimicrobial and endotoxin-neutralizing activities. These molecules show a broad spectrum of activity against various pathogens (including bacteria, viruses, fungi etc.) These compounds can be used alone or in combination therapy with conventional antibiotics or antiendo toxic agents.

[0013] US 5,736,574 discloses antimicrobial immixtures comprising at least one antimicrobial hydrolipid and / or lipid and an antimicrobially synergistically effective amount of at least one glyceryl mono alkyl ether are well suited for formulation, as preservatives, into a wide variety of pharmaceutical / cosmetic compositions.

[0014] US 5,043,176 discloses an antimicrobial composition composed of an antimicrobial polypeptide and a hypothiocyanate component. Synergistic activity is seen when the composition is applied at between about 30 and 40 degrees Centigrade at a pH between about 3 and about 5 for a contact time of 10 to 20 min. The composition is useful against gram negative bacteria such as Salmonella. A preferred composition is nisin, lactoperoxidase, thiocyanate and hydrogen peroxide.

[0015] US 2013 / 0210705A1 discloses antimicrobial metallodrugs comprising an antimicrobial peptide (“AMP”) and / or an antibiotic covalently bound to a metal binding moiety. These metallodrugs combine a metal binding domain which typically catalyzes oxido-reductase chemistry or acts as a Lewis-Acid catalyst, with a member of a diverse class of antimicrobial agents currently validated in preclinical and clinical settings for the treatment of a broad spectrum of pathogenic organisms.

[0016] 2795 / MUMNP / 2011 is directed to antimicrobial agents against Gram negative bacteria, in particular to fusion proteins composed of an enzyme having the activity of degrading the cell wall of Gram-negative bacteria and an additional peptide stretch fused to the enzyme on the N- or C-terminus and pharmaceutical or cosmetic compositions comprising said fusion protein. The peptide is an antimicrobial peptide including indolicidin and cathelicidin.

[0017] Antimicrobial properties are inherent to AMPs. However, the antimicrobial property cited in the prior art is mainly related to determination of Minimum Inhibitory Concentration (MIC) & Minimum Bactericidal Concentration (MBC). There are few reports of determination of log reduction for AMPs. However, the first time point for measuring the log reduction is 30 minutes. For hand hygiene applications, the relevant time points are 1 minute and 5 minutes. For AMPs such data is not cited in the prior art.

[0018] Further, AMPs cited in prior art are tested in conditions that do not mimic real-life like scenario with respect to hand hygiene standard requirements. For example, mimicking soil conditions by testing in the presence of protein, Bovine Serum Albumin (BSA), and testing at temperature such as 20°C and for 1 minute / 5 minute as the time for testing depending on the applications. The British Standard EN1276 has laid out stringent protocol for antibacterial efficacy testing of the active system / formulation for hand hygiene applications. There are no prior art report of AMP containing active system / composition achieving >5 log kill in 1 minute versus both Gram positive and Gram negative test organisms as per the British Standard BS EN 1276:2009.

[0019] There is a need for a composition of AMP with specific surfactant that demonstrate >5 log kill in 1 minute against both Gram positive and Gram negative bacteria in presence of rigorous conditions that simulate hand hygiene conditions.

[0020] Object of the Invention

[0021] It is an object of the present invention to provide an antimicrobial composition comprising AMP and a non-ionic surfactant. It is also an object of the present invention to provide an antimicrobial composition comprising AMP and a non-ionic surfactant showing at least 3 log kill in 1 minute against both Gram positive and Gram negative bacteria.

[0022] It is also an object of the present invention to provide an antimicrobial composition comprising LL-37 peptide fragment FK-16 (Cl) and a non-ionic surfactant.

[0023] It is a further object to provide formulations comprising the antimicrobial composition comprising LL-37 peptide fragment FK-16 (Cl) and a non-ionic surfactant for topical use.

[0024] Summary of the Invention

[0025] In one aspect of the present invention there is provided an antimicrobial composition comprising an effective concentration of cathelicidin peptide fragment (FK16) as set forth in SEQ ID 1, or salts and derivatives thereof and an effective concentration of at least one non-ionic surfactant, such that the antimicrobial composition achieves a log-kill of at least 3, after a contact time of about 1 min against at least one microbe.

[0026] In another aspect of the present invention there is provided an antimicrobial composition comprising an effective concentration of cathelicidin peptide fragment (FK16) as set forth in SEQ ID 1, or salts and derivatives thereof, an effective concentration of at least one non-ionic surfactant selected from Decyl-Glucoside, Caprylyl / Capryl Glucoside, Ethyl Hexanol Ethoxylated Propoxylate(Ecosurf™ EH-9), or combinations thereof, and an effective concentration of at least one chelating agent selected from disodium EDTA and organic acid like malic acid, citric acid, GLDA and PESA such that the antimicrobial composition achieves a log-kill of at least 3, after a contact time of about 1 min against at least one microbe.

[0027] In a further aspect of the present invention there is provided an alcohol-free wet wipe formulation comprising the antimicrobial composition described herein. In yet another aspect of the present invention there is provided a method of disinfecting a surface, the method comprising the steps of: providing an alcohol- free wet wipe as described herein; and wiping the surface with the wet wipe.

[0028] In another aspect of the present invention there is provided an alcohol-free deodorant formulation comprising the antimicrobial composition as described herein.

[0029] In another aspect of the present invention there is provided a method of reducing body odour, the method comprising the steps of: providing an alcohol-free deodorant as described herein and applying the deodorant to the skin in mist spray format.

[0030] Brief Description of Accompanying drawings

[0031] Figure 1 illustrates HPLC chromatogram of Cl peptide

[0032] Detailed Description of the invention

[0033] The present invention provides a synergistic combination of Antimicrobial Peptide (AMP) and non-ionic surfactant for hygiene application.

[0034] The following description with reference to the accompanying drawings is provided to assist in a comprehensive understanding of exemplary embodiments of the invention. It includes various specific details to assist in that understanding but these are to be regarded as merely exemplary.

[0035] Accordingly, those of ordinary skill in the art will recognize that various changes and modifications of the embodiments described herein can be made departing from the scope of the invention. In addition, descriptions of well known functions and constructions are omitted for clarity and conciseness. The terms and words used in the following description and claims are not limited to the bibliographical meanings, but, are merely used by the inventor to enable a clear and consistent understanding of the invention. Accordingly, it should be apparent to those skilled in the art that the following description of exemplary embodiments of the present invention are provided for illustration purpose only and not for purpose of limiting the scope of the invention as defined by the appended claims and their equivalents.

[0036] It is to be understood that the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise.

[0037] Features that are described and / or illustrated with respect to one embodiment may be used in the same way or in a similar way in one or more other embodiments and / or in combination with or instead of the features of the other embodiments.

[0038] It should be emphasized that the term “comprises / comprising” when used in this specification is taken to specify the presence of stated features, steps or components but does not preclude the presence or addition of one or more other features, steps, components or groups thereof.

[0039] The term "peptide" as used herein refers to short polypeptides consisting from 2 to 50 amino acid residues, “Polypeptides” and “proteins” comprise more than 50 amino acid residues. A peptide may have a specific function. A peptide can be a naturally occurring peptide or a synthetically designed and produced peptide. The peptide can be, for example, derived or removed from a native protein by enzymatic or chemical cleavage, or can be prepared using conventional peptide synthesis techniques (e.g., solid phase synthesis) or molecular biology techniques (Sambrook, 10 J. et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989)). The term, "antimicrobial peptide" (AMP) as used herein refers to any peptide that has microbiocidal and / or microbio static activity. Thus, the term "antimicrobial peptide" as used herein refers in particular to any peptide having anti-bacterial, antifungal, anti-mycotic, antiparasitic, anti-protozoal, anti-viral, anti-infectious, antiinfective and / or germicidal, algicidal, amoebicidal, microbiocidal, bactericidal, fungicidal, parasiticidal, protozoacidal properties.

[0040] The term “derivatives” as used herein includes peptide analogues, peptide fragments and modified peptides. The derivatives may be a sequence that has at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 1 of present invention.

[0041] In tune with growing environmental concern, some of the embodiments of the present invention make use of environment friendly and biodegradable organic acid based chelating agents.

[0042] The term “log kill / log reduction” as used herein denotes the number of cells killed in a test. If a log reduction is a whole integer, then its numerical value equals the number of nines in the percent reduction figure. For e.g. 1 log reduction = 90% reduction, 2 log reduction = 99% reduction, 3 log reduction = 99.9% reduction, 4 log reduction = 99.99% reduction, 5 log reduction = 99.999% reduction, 6 log reduction = 99.9999% reduction.

[0043] The term “minimum inhibitory concentration (MIC)” of an antimicrobial peptide is the lowest concentration of peptide that completely inhibits growth of the organism. In general, lower MIC values are preferred.

[0044] The term “Hygiene” as used herein means a series of practices performed to preserve health. According to the World Health Organization (WHO), "Hygiene refers to conditions and practices that help to maintain health and prevent the spread of diseases." The term “peptide fragment” as used herein means functional motifs / shorter fragments which retain the biological activity of the full-length peptide (in the present invention LL-37).

[0045] The term "skin" as used herein means the entire epidermis of mammals including human.

[0046] The present invention relates to an antimicrobial composition comprising an antimicrobial peptide (AMP) and one or more non-ionic surfactants.

[0047] The invention provides benefits of reducing pathogenic microbial load on skin and hence, helps in skin hygiene and sanitization.

[0048] The AMP active domains have potential to be used in personal hygiene products due to their ability to kill pathogenic microorganisms. The AMPs act on the bacterial membrane instead of single metabolic event. There will be less pressure on developing antimicrobial resistance to the two AMP actives in the present application. The AMPs act by disrupting the cell membrane, not on single step. Studies have demonstrated that the resistance level provided by point mutations and gene amplifications is very low in case of AMPs.

[0049] The present invention provides synergistic antimicrobial combinations comprising an effective concentration of antimicrobial peptide (AMP) and an effective concentration of non-ionic surfactant.

[0050] In an embodiment of the present invention there is provided an antimicrobial composition comprising an effective concentration of a cathelicidin LL-37 peptide fragment and an effective concentration of non-ionic surfactant as active ingredients. LL-37 peptide fragment FK-16 (Cl) as set forth in SEQ ID No. 1 corresponding to residues 17-32 retains antibacterial and antitumor effects. [Reference: Ren SX, Shen J, Cheng AS, et al. FK-16 derived from the anticancer peptide LL-37 induces caspase-independent apoptosis and autophagic cell death in colon cancer cells [published correction appears in PLoS One. 2015;10(6):e0131750]. PLoS One. 15 2013;8(5):e63641. Published 2013 May 20. doi: 10.1371 / journal.pone.0063641]

[0051] FK16 or Cl kills a range of pathogens in both in vitro and in vivo systems. FK16 is capable of killing a variety of pathogens and do not elicit caustic immunologic responses and host tissue toxicity. FK16 covers 42% of full length parent, native human Cathelicidin LL-37 peptide.

[0052] Typically, synthetic peptides are produced as salts of trifluroacetate (TFA), acetate, chloride, lactate etc,. However, the peptide sequence is the active principle that is linked to its efficacy, that is, antimicrobial activity. Hence, all salt forms of the peptide (TFA, acetate etc) will have similar activities.

[0053] The present invention provides an antimicrobial composition comprising an effective concentration of cathelicidin peptide fragment (FK16) as set forth in SEQ ID 1, or salts and derivatives thereof and an effective concentration of at least one non-ionic surfactant, such that the antimicrobial composition achieves a log-kill of at least 3, after a contact time of about 1 min against at least one microbe.

[0054] The antimicrobial composition as disclosed herein comprises FK16 (Cl) peptide at a concentration of 0.01%-0.15% with respect to the total weight of the composition. In an embodiment, the FK16 (Cl) peptide is present at a concentration selected from 0.0125% or 0.02% or 0.025% or 0.05% or 0.1% or 0.15%. In other embodiment, the concentration of FK16 (Cl) peptide can be any value within the range. In a preferred embodiment, the antimicrobial composition as disclosed herein comprises FK16 (Cl) peptide at a concentration of 0.0125%-0.05% with respect to the total weight of the composition. In a most preferred embodiment, FK16 (Cl) peptide is present at a concentration of 0.025% with respect to the total weight of the composition.

[0055] In an embodiment, at least one non-ionic surfactant is selected from the group of Alkyl-Poly Glucosides (APG). Preferably, the Alkyl-Poly Glucoside (APG) is selected from Decyl-Glucoside, Lauryl Glucoside, Capryl glucoside, caprylyl / capryl glucoside, Arachidyl glucoside or combinations thereof.

[0056] In a preferred embodiment, Alkyl-Poly Glucoside (APG) is Decyl-Glucoside. The antimicrobial composition as disclosed herein comprises Decyl-Glucoside at a concentration of 0.0625% to 0.5% with respect to the total weight of the composition.

[0057] In another embodiment, the at least one non-ionic surfactant is selected from Secondary alcohol ethoxylates. Preferably, the Secondary alcohol ethoxylates is Ethyl Hexanol Ethoxylated Propoxylate (Ethylene oxide-propylene oxide copolymer mono(2-ethylhexyl) ether) (Ecosurf™ EH-9).

[0058] The antimicrobial composition as disclosed herein comprises Ethyl Hexanol Ethoxylated Propoxylate at a concentration of 0.0625% to 4% with respect to the total weight of the composition.

[0059] In an embodiment, the non-ionic surfactant is a combination of both Alkyl-Poly Glucoside and Secondary alcohol ethoxylates. In a preferred embodiment, the non- ionic surfactant is Decyl-Glucoside and Ethyl Hexanol Ethoxylated Propoxylate. The antimicrobial composition comprises Decyl-Glucoside to Ethyl Hexanol Ethoxylated Propoxylate in a ratio ranging from 1.6: 1 to 2.4: 1.

[0060] In an embodiment, the antimicrobial peptide and the non-ionic surfactant are combined at a ratio ranging from 0.003125: 1 to 0.8: 1. In one embodiment, the antimicrobial composition comprises FK16 (Cl) peptide and Decyl-Glucoside. The Cl peptide is present in a concentration range from 0.0125% to 0.05% and Decyl-Glucoside is present in a concentration range from 0.0625% to 0.5%. The ratio of FK16 (Cl) peptide and Decyl-Glucoside in the antimicrobial composition ranges from 0.025:1 to 0.8:1 by weight.

[0061] In an embodiment, the antimicrobial composition comprises 0.0125% by weight of FK16 (Cl) peptide and 0.0625% by weight of Decyl-Glucoside. In an embodiment, the antimicrobial composition comprises 0.0125% by weight of FK16 (Cl) peptide and 0.125% by weight of Decyl-Glucoside. In an embodiment, the antimicrobial composition comprises 0.0125% by weight of FK16 (Cl) peptide and 0.25% by weight of Decyl-Glucoside. In an embodiment, the antimicrobial composition comprises 0.0125% by weight of FK16 (Cl) peptide and 0.5% by weight of Decyl- Glucoside.

[0062] In an embodiment, the antimicrobial composition comprises 0.025% by weight of FK16 (Cl) peptide and 0.0625% by weight of Decyl-Glucoside. In an embodiment, the antimicrobial composition comprises 0.025% by weight of FK16 (Cl) peptide and 0.125% by weight of Decyl-Glucoside. In an embodiment, the antimicrobial composition comprises 0.025% by weight of FK16 (Cl) peptide and 0.25% by weight of Decyl-Glucoside. In an embodiment, the antimicrobial composition comprises 0.025% by weight of FK16 (Cl) peptide and 0.5% by weight of Decyl- Glucoside.

[0063] In an embodiment, the antimicrobial composition comprises 0.05% by weight of FK16 (Cl) peptide and 0.0625% by weight of Decyl-Glucoside. In an embodiment, the antimicrobial composition comprises 0.05% by weight of FK16 (Cl) peptide and 0.125% by weight of Decyl-Glucoside. In an embodiment, the antimicrobial composition comprises 0.05% by weight of FK16 (Cl) peptide and 0.25% by weight of Decyl-Glucoside. In an embodiment, the antimicrobial composition comprises 0.05% by weight of FK16 (Cl) peptide and 0.5% by weight of Decyl- Glucoside.

[0064] In another embodiment, the antimicrobial composition comprises FK16 (Cl) peptide and Ethyl Hexanol Ethoxylated Propoxylate. The C 1 peptide is present in a concentration range from 0.0125% to 0.05% and Ethyl Hexanol Ethoxylated Propoxylate is present in a concentration range from 0.0625% to 4%. The ratio of FK16 (Cl) peptide and Ethyl Hexanol Ethoxylated Propoxylate in the antimicrobial composition ranges from 0.003125: 1 - 0.8: 1 by weight.

[0065] In another embodiment, the antimicrobial composition comprises 0.0125% by weight of FK16 (Cl) peptide and 0.0625% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.0125% by weight of FK16 (Cl) peptide and 0.125% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.0125% by weight of FK16 (Cl) peptide and 0.25% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.0125% by weight of FK16 (Cl) peptide and 0.5% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.0125% by weight of FK16 (Cl) peptide and 1% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.0125% by weight of FK16 (Cl) peptide and 4% by weight of Ethyl Hexanol Ethoxylated Propoxylate.

[0066] In another embodiment, the antimicrobial composition comprises 0.025% by weight of FK16 (Cl) peptide and 0.0625% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.025% by weight of FK16 (Cl) peptide and 0.125% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.025% by weight of FK16 (Cl) peptide and 0.25% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.025% by weight of FK16 (Cl) peptide and 0.5% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.025% by weight of FK16 (Cl) peptide and 1% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.025% by weight of FK16 (Cl) peptide and 4% by weight of Ethyl Hexanol Ethoxylated Propoxylate.

[0067] In another embodiment, the antimicrobial composition comprises 0.05% by weight of FK16 (Cl) peptide and 0.0625% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.05% by weight of FK16 (Cl) peptide and 0.125% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.05% by weight of FK16 (Cl) peptide and 0.25% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.05% by weight of FK16 (Cl) peptide and 0.5% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.05% by weight of FK16 (Cl) peptide and 1% by weight of Ethyl Hexanol Ethoxylated Propoxylate. In another embodiment, the antimicrobial composition comprises 0.05% by weight of FK16 (Cl) peptide and 4% by weight of Ethyl Hexanol Ethoxylated Propoxylate.

[0068] In yet another embodiment, the antimicrobial composition comprises FK16 (Cl) peptide and Decyl-Glucoside and Ethyl Hexanol Ethoxylated Propoxylate. In yet another embodiment, the antimicrobial composition comprises 0.025% by weight FK16 (Cl) peptide and 0.20% by weight Decyl-Glucoside and 0.125% by weight Ethyl Hexanol Ethoxylated Propoxylate.

[0069] The antimicrobial composition comprises Decyl-Glucoside to Ethyl Hexanol

[0070] Ethoxylated Propoxylate in a ratio ranging from 1.6:1 to 2.4: 1 The present invention provides a synergistic combination of human cathelicidin fragment- AMP (FK-16) and non-ionic surfactant that synergistically enhances bactericidal efficacy. In 1 minute contact kill experiment (in presence of 0.3% BSA (final concentration)], the composition of peptide (Cl) and non-ionic surfactant lead to higher log reduction of at least 3 against Gram negative bacteria Pseudomonas aeruginosa, Escherichia coli and Gram positive bacteria Staphylococcus aureus, Enterococcus hirae.

[0071] In one of the embodiments, the antimicrobial composition further comprises a chelating agent. The chelating agent can be selected from disodium EDTA and environment friendly and biodegradable ingredient like organic acids, such as malic acid, citric acid, GLDA (Glutamic acid Di Acetate) and PESA (Polyepoxy succinic acid). In one embodiment, the chelating agent is disodium EDTA. In another embodiment, the chelating agent is malic acid.

[0072] The antimicrobial composition as disclosed herein comprises disodium EDTA at a concentration of 0.1 to 0.4% with respect to the total weight of the composition. In a preferred embodiment, the concentration of EDTA can be selected from 0.1% or 0.15% or 0.2% or 0.25% or 0.3% or 0.35% with respect to the total weight of the composition.

[0073] The antimicrobial composition as disclosed herein comprises malic acid at a concentration in the range of 0.01 to 2% with respect to the total weight of the composition. In a preferred embodiment, the concentration of malic acid can be selected from 0.05% or 0.15% with respect to the total weight of the composition.

[0074] Accordingly, in one embodiment, the present invention provides an antimicrobial composition comprising an effective concentration of cathelicidin peptide fragment (FK16) as set forth in SEQ ID 1, or salts and derivatives thereof, an effective concentration of at least one non-ionic surfactant, and an effective concentration of at least one chelating agent. In a preferred embodiment, the antimicrobial composition comprising an effective concentration of cathelicidin peptide fragment (FK16) as set forth in SEQ ID 1, or salts and derivatives thereof, an effective concentration of at least one non-ionic surfactant selected from Decyl-Glucoside, Ethyl Hexanol Ethoxylated Propoxylate, or combinations thereof, and an effective concentration of at least one chelating agent selected from disodium EDTA, malic acid, citric, GLDA and PESA acid such that the antimicrobial composition achieves a log-kill of at least 3, after a contact time of about 1 min against at least one microbe.

[0075] In one embodiment, the antimicrobial composition comprises FK16 (Cl) peptide, Decyl-Glucoside, and disodium EDTA. In one embodiment, the antimicrobial composition comprises 0.025% by weight FK16 (Cl) peptide, 0.25% by weight Decyl-Glucoside and 0.25% by weight disodium EDTA. In one embodiment, the antimicrobial composition comprises 0.025% by weight FK16 (Cl) peptide, 0.5% by weight Decyl-Glucoside and 0.25% by weight disodium EDTA. In one embodiment, the antimicrobial composition comprises 0.025% by weight FK16 (Cl) peptide, 0.125% by weight Decyl-Glucoside and 0.1% by weight disodium EDTA. In one embodiment, the antimicrobial composition comprises 0.025% by weight FK16 (Cl) peptide, 0.30% by weight Decyl-Glucoside and 0.20% by weight disodium EDTA.

[0076] In another embodiment, the antimicrobial composition comprises FK16 (Cl) peptide, Decyl-Glucoside, and malic acid. In another embodiment, the antimicrobial composition comprises 0.025% by weight FK16 (Cl) peptide, 0.5% by weight Decyl-Glucoside, and 0.05% by weight malic acid. In another embodiment, the antimicrobial composition comprises 0.025% by weight FK16 (Cl) peptide, 0.5% by weight Decyl-Glucoside, and 0.15% by weight malic acid.

[0077] In yet another embodiment, the antimicrobial composition comprises FK16 (Cl) peptide, Decyl-Glucoside, disodium EDTA and malic acid. In yet another embodiment, the antimicrobial composition comprises 0.025% by weight FK16 (Cl) peptide, 0.5% by weight Decyl-Glucoside, 0.4% by weight disodium EDTA and 0.15% by weight malic acid.

[0078] In another embodiment, the antimicrobial composition comprises FK16 (Cl) peptide, Ethyl Hexanol Ethoxylated Propoxylate and disodium EDTA. In another embodiment, the antimicrobial composition comprises 0.025% by weight FK16 (Cl) peptide, 0.5% by weight Ethyl Hexanol Ethoxylated Propoxylate and 0.25% by weight disodium EDTA. In another embodiment, the antimicrobial composition comprises 0.025% by weight FK16 (Cl) peptide, 0.25% by weight Ethyl Hexanol Ethoxylated Propoxylate and 0.1% by weight disodium EDTA.

[0079] In yet another embodiment, the antimicrobial composition comprises FK16 (Cl) peptide, Ethyl Hexanol Ethoxylated Propoxylate and malic acid.

[0080] In a further embodiment, the antimicrobial composition comprises FK16 (Cl) peptide, Decyl-Glucoside, Ethyl Hexanol Ethoxylated Propoxylate and disodium EDTA. In a further embodiment, the antimicrobial composition comprises 0.025% by weight FK16 (Cl) peptide, 0.30% by weight Decyl-Glucoside, 0.125% by weight Ethyl Hexanol Ethoxylated Propoxylate and 0.20% by weight disodium EDTA.

[0081] In another embodiment, the antimicrobial composition comprises FK16 (Cl) peptide, Decyl-Glucoside, Ethyl Hexanol Ethoxylated Propoxylate and malic acid.

[0082] The antimicrobial compositions of the present invention may further comprise cosmetically acceptable excipients selected from, emulsifiers, solvents, rheology modifying agents, filler s / thickening agents, structurants, emollients, diluents, humectants, feel enhancers, preservatives and fragrances.

[0083] Thickening agents include but are not limited to hydrogels, PEG (polyethyelene glycol), polyacrylic acid, hydroxypropyl methylcellulose (HPMC), Bentone, Natural or synthetic gum. Emulsifying agents include but are not limited to derivatives of PEG (Polyethylene Glycol), Polysorbate. Diluents include but are not limited to water or ethyl alcohol. Rheology modifying agents include but are not limited to hydrophilic polymers. Structurants include but are not limited to hydrogenated vegetable oil, hydrogenated castor oil, fatty acids, beeswax, paraffin wax. Humectants include but are not limited to glycerine and derivatives. Emollients include but are not limited to fatty acids, fatty alcohol esters, hydrocarbons, mineral oils, polyorganosiloxane. Fragrances include but are not limited to give woody notes, floral notes, oriental notes and fresh notes. Preservatives include but are not limited to essential oils, organic acids and synthetic agents.

[0084] The antimicrobial compositions of the present invention may be in the form of wipes, hand wash, deodorants. Other forms include but not limited to sanitizer, body wash, soap, shampoo and gels, wound healing cream / lotion and surface cleaner.

[0085] The present invention provides an alcohol-free wet wipe formulation comprising an effective concentration of cathelicidin peptide fragment (FK 16 / CI) as set forth in SEQ ID 1 and an effective concentration of least one non-ionic surfactant.

[0086] The wet wipe formulation further comprises a chelating agent.

[0087] The non-ionic surfactant is selected from Decyl-Glucoside, Ethyl Hexanol Ethoxylated Propoxylate, or combinations thereof. The chelating agent is selected from disodium EDTA or malic acid, or citric acid or GLDA or PESA.

[0088] The wet wipe formulation further comprises of at least one excipient selected from the group consisting of preservative, pH modifier, humectant, fragrance, fragrance solubilizer etc. The wet-wipe has a pH ranging from 5 to 6 (pH 5.5) (skin friendly pH range) and is characterized by log kill of 5 after a contact time of about 1 minute against Pseudomonas aeruginosa, ATCC 15442, Escherichia coli ATCC 10536, Staphylococcus aureus ATCC 6538 and Enterococcus hirae ATCC 10541.

[0089] The present invention provides a method of disinfecting a surface, the method comprising the steps of: providing an alcohol-free wet wipe as described herein; and wiping the surface with the wet wipe.

[0090] The present invention provides an alcohol-free deodorant formulation comprising an effective concentration of cathelicidin peptide fragment (FK 16 / CI) as set forth in SEQ ID 1, an effective concentration of least one non-ionic surfactant and a chelating agent.

[0091] The non-ionic surfactant is selected from Decyl-Glucoside, Ethyl Hexanol Ethoxylated Propoxylate, or combinations thereof. The chelating agent is disodium EDTA, or malic acid or citric acid or GLDA or PESA

[0092] The alcohol-free deodorant formulation further comprises of at least one excipient selected from the group consisting of preservative, fragrance, fragrance solubilizer etc.

[0093] In one embodiment, the alcohol-free deodorant formulation is in the form of a mist spray format without fragrance. In another embodiment, the alcohol-free deodorant formulation is in the form of a mist spray format with fragrance.

[0094] The present invention also provides a method of reducing body odour, the method comprising the steps of: providing an alcohol-free deodorant as described herein and applying the deodorant to the skin in mist spray format. The present invention provides a synergistic combination of an antimicrobial peptide and a surfactant for the manufacture of various compositions for disinfecting surfaces or reducing body odour, wherein the combination is active against both gram-positive and gram-negative bacteria.

[0095] The present invention resides in the combination of the specific antimicrobial peptide with the particular surfactants and also chelating agents. Not all antimicrobial peptide is compatible with a specific surfactant and similarly, not all surfactant is compatible with a particular anti-microbial peptide. The anti-microbial peptides are highly sensitive moieties extremely susceptible to slightest change of parameters (HLB value of the surfactant, ionic or non-ionic nature of the surfactant and even mere chain length of the surfactant). Similarly, all chelating agents are not compatible with a particular antimicrobial peptide. Therefore, the particular combinations claimed in the present invention are judicious combinations leading to the inventive merit.

[0096] In prior art patents, AMPs are listed as anti-infective agents and AMP and antibiotic synergy is demonstrated but synergy is not demonstrated in terms of significant log kill i.e >5 log kill ( 99.999% reduction) of microorganisms in 1 minute. The prior arts also do not demonstrate synergy by antimicrobial peptide based active system against broad spectrum of bacteria including pathogenic Gram positive and Gram negative bacteria.

[0097] >5 log kill is required for applications like hand wash and sanitizers. 3 log kill is considered as bactericidal based on definition. 3 log reduction in 1 minute could be considered for certain applications as well.

[0098] The experimental conditions used in prior art do not mimic conditions required for hand hygiene applications; For example, 1 minute contact time, using bovine serum albumin (BSA) as interfering agent. In the present invention, the combination of AMP (peptide Cl and surfactant) demonstrated synergy in antimicrobial efficacy than at tested individual concentrations. The AMP + surfactant composition work synergistically at lower concentrations providing 99.999% reduction against Gram positive and Gram negative bacteria under rigorous, challenging testing conditions. The typical cost of chemical synthesis of peptide is on higher side. In the present invention, there is a significant reduction of peptide concentration achieved in presence of synergistic active which is economical.

[0099] The present invention is now being illustrated by way of non-limiting examples. The examples are intended to be purely exemplary of the invention and should therefore not be considered to limit the invention in any way. Efforts have been made to ensure accuracy with respect to numbers used, but some experimental errors and deviations should be accounted for.

[0100] Materials and methods:

[0101] Description of the components of the invention

[0102] 1. Peptide Cl (FK-16): The 16 amino acid peptide with sequence FKRIVQRIKDFLRNLV was custom synthesized from USV Pvt. Etd, India at >98% purity based on HPEC profile. The HPEC chromatogram is enclosed (Figure 1). The molecular weight of the peptide is 2045 Daltons.

[0103] 2. Decyl Glucoside (58846-77-8): Plantacare® 2000 UP from BASF

[0104] 3. Secondary alcohol ethoxylate / Ethyl Hexanol Ethoxylated Propoxylate / Ethylene oxide -propylene oxide copolymer mono(2-ethylhexyl) ether (CAS NO: 64366-70-7): (ECOSURF™ EH-9) from Sigma STS0006

[0105] 4. PEG-40 Hydrogenated Castor Oil (CAS Number. 61788-85-0): CREMOPHOR® CO 40 from BASF

[0106] 5. DL malic acid: CAS No. 6915-15-7 from Sigma -Aldrich (USA) Catalogue No. 240176

[0107] 6. Citric acid - BDH GPR™ 279844E, VWR International Limited 7. Disodium EDTA: CAS No. 6381-92-7 from Sigma-Aldrich (USA) Catalogue No. E5134

[0108] 8. Caprylyl / Capryl Glucoside (68515-73-1): ORAMIX™ CG 110

[0109] Microorganism :

[0110] The microorganisms used in the study are:

[0111] 1. Pseudomonas aeruginosa, ATCC 15442

[0112] 2. Escherichia coli ATCC 10536

[0113] 3. Staphylococcus aureus ATCC 6538

[0114] 4. Enterococcus hirae ATCC 10541

[0115] 5. Staphylococcus epidermidis MTCC 435

[0116] 6. Corynebacterium striatum ATCC 6940

[0117] The glycerol stock of these bacteria was stored at -80°C. The culture was maintained on Tryptone soya agar (TSA) and sub-cultured every two weeks.

[0118] Antimicrobial activity Assay to determine Microbicidal Efficacy:

[0119] Antimicrobial activity to calculate the level of killing achieved by the test samples was determined using quantitative suspension test (contact kill) following the guidelines specified by EN 1276 Standard at 20°C using Dilution-neutralization method. Briefly, the cells were contacted with test samples for predefined time interval (generally 1 minute ±5 seconds or 5 minutes ±5 seconds) at 20°C, the antimicrobial activities of samples stopped dilution-neutralization method using a neutralizer solution & estimating the number of surviving bacteria using TSA plates.

[0120] Bovine serum albumin (heat shock fraction, pH 5.2, >96%) procured from Sigma Chemical Company (Catalog no A8022) was used as interfering substance in the assay to simulate soiling conditions at dirty levels as defined by EN 1276 Standard. Tryptone Soya Agar (TSA), Tryptone soya broth (TSB) and other chemicals were procured from HiMedia Laboratories, Mumbai, India. Detailed Method: Colonies of bacteria from the TSA plate were suspended in diluent to prepare bacterial cell suspension of about 3 X 108CFU / ml. Test samples, bacterial suspension and 3% BSA solution were equilibrated at 20°C for 10 min in a dry bath (Eppendorf Thermomixer comfort). To 100 pl of bacterial suspension, 100 pl of BSA was added to simulate soiling condition. Within 5 minutes of preparation, this 200 pl bacteria was added to 800 pl of test samples & immediately mixed using a vortex mixer. At predefined contact time (generally 1 minute), the action of antimicrobial was stopped using dilution neutralization method. For this, an aliquot of assay content was added to 9X volumes of neutralizer solution (Polysorbate 80, 30g / l; Saponin, 30 g / 1; Lecithin, 3 g / 1 in 250mM potassium phosphate buffer; pH 7.2) and left for 5 minutes. The surviving bacterial counts in the neutralized samples were estimated using 10-fold serial dilutions on TSA agar plates and incubated 37°C for 24 hours. The colonies developed on TSA plates were counted and antimicrobial activity was calculated. Activity was evaluated in terms of logio reduction factor (Log kill or Log reduction) that was calculated as the difference between logarithms of CFU / ml before and after exposure to test samples. A log kill / reduction of 1 translates to 90% killing of bacteria. Similarly, log kill / reduction of 2, 3, 4 & 5 translates to 99%, 99.9%, 99.99% and 99.999% bacterial killing respectively.

[0121] Results & Discussion:

[0122] Example 1: Antimicrobial efficacy of Cl:

[0123] Antimicrobial peptide Cl shows antimicrobial activity against Escherichia coli with >4 log reduction of bacterial counts in 1 minute (Table 1) at concentrations of 0.01% and above. However, it failed to kill Enterococcus hirae, even at concentrations as high as 0.15%. E. hirae is one of the mandatory bacteria specified by EN 1276 standard for demonstrating sanitization and disinfection activity of chemicals. The activity against E. hirae improved with time, yet it was still considerably lower at 2.6 log reduction (Table 2). Table 1. Antimicrobial efficacy of Antimicrobial Peptide Cl against Escherichia coli & Enterococcus hirae estimated in terms of log reduction of its viable cell counts achieved after 1 minute contact time.

[0124] Table 2. Antimicrobial efficacy of Antimicrobial Peptide Cl against Enterococcus hirae estimated in terms of log reduction of its viable cell counts achieved at different contact time.

[0125] Example 2: Antimicrobial efficacy of Cl & Surfactants combinations

[0126] Synthesis of antimicrobial peptides is a complex process and requires considerable cost. The higher costs associated with peptide makes it, uneconomical for its usage in various antimicrobial applications. To develop synergistic combination of Cl for decreasing its usage level and enhancing antimicrobial activity, we evaluated its combinations with various surfactants, which are core groups of molecules used in personal care applications.

[0127] Table 3. Antimicrobial efficacy of combinations of Antimicrobial Peptide Cl and various surfactants against Escherichia coli and Enterococcus hirae estimated in terms of log reduction of its viable cell counts achieved after contact time of 1 minute.

[0128] The combination of AMP Cl (0.025%) and various surfactants (0.4% & 4%) were tested against E. coli & E. hirae. Most of the surfactants completely disrupted the activity of Cl as demonstrated by loss of antimicrobial activity against E. coli compared to the killing by C 1 alone (Table 3). Only combinations of C 1 with Decyl

[0129] Glucoside and Ethyl Hexanol Ethoxylated Propoxylate exhibited antimicrobial activity. These two combinations with non-ionic surfactants, surprisingly demonstrated activity against E. hirae particularly, that stands out considering that Polysorbate 20 and Polysorbate 80, other non-ionic surfactants tested here, completely disrupted activity of C 1. Decyl glucoside and Capryyl / capryl Glucoside belong to Alkyl polyglycoside (APG) group of surfactants while Ethyl Hexanol Ethoxylated Propoxylate is an Alcohol Ethoxylate. When tested alone, Decyl Glucoside, Capryyl / capryl Glucoside and Ethyl Hexanol Ethoxylated Propoxylate show very little antimicrobial activity during same contact time of 1 minute (Table 4).

[0130] Table 4. Antimicrobial efficacy of non-ionic surfactants against Escherichia coli and Enterococcus hirae after contact time of 1 minute Example 3A: Antimicrobial efficacy of Cl and Decyl glucoside combinations

[0131] Various combinations of AMP Cl and Decyl glucosides covering Cl: Decyl Glucoside ratios from 0.0016:1 to 1.6:1 were tested against E. hirae. Table 5 summarizes the antimicrobial activities of all these combinations. Cl alone (0.00625% to 0.05%) or Decyl Glucoside alone (0.003125% to 4%) do not exhibit antimicrobial activity. However, various combinations covering Cl levels of 0.0125%-0.05% with 0.0625% - 0.5% Decyl Glucoside provided >3 log reduction in E. hirae counts in contact kill assay demonstrating their strong synergistic antimicrobial effect. The synergy was defined as enhancement in killing of bacteria by at least 1 log (i.e. 10-fold in percentage value) by a combination compared to additive effect of individual actives that are part of the combination. Further, the combination needs to exhibit at least 3 log reduction of viable counts of bacteria to be considered as strong antimicrobial for its utility.

[0132] The Cl to Decyl Glucoside ratios ranging from 0.025: 1 - 0.8: 1 exhibits strong synergy in their antimicrobial efficacy that kills E. hirae effectively.

[0133] At Decyl Glucoside levels above 0.5%, the combinations showed reduced activity. Nonetheless, this activity was still higher than individual activity of Cl or Decyl Glucoside.

[0134] Table 5A. Antimicrobial efficacy: log reduction achieved with various combinations of Cl + Decyl glucoside after 1 minute contact time at 20°C vs E. hirae

[0135] Example 3B: Antimicrobial efficacy of Cl and Caprylyl / Capryl glucoside (CG) combinations

[0136] Further activity of Cl with various combinations of Caprylyl / Capryl glucoside (CG) were tested for antimicrobial activity on various bacteria like E. hirae, E. coli It was found that for combinations of Cl with Caprylyl / Capryl glucoside (CG) in ratios 0.0125: 0.25 and 0.0125:2 provide log kill value of more than 5 in one minute.

[0137] The action of the said combinations is evident from the following table 5B.

[0138] Table 5B. Antimicrobial efficacy: log reduction achieved with various combinations of Cl with Caprylyl / Capryl glucoside (CG), after 1 minute contact time at 20°C

[0139] Example 4A: Antimicrobial efficacy of Cl and Ecosurf™ EH-9 combinations Various combinations of AMP Cl and Ecosurf™ EH-9 covering Cl : Ethyl Hexanol Ethoxylated Propoxylate (Ecosurf™ EH-9 ) ratios from 0.0016:1 to 1.6:1 were tested against E. hirae. Table 6A summarizes the antimicrobial activities of all these combinations. Cl alone (0.00625% to 0.05%) or Ethyl Hexanol Ethoxylated Propoxylate alone (0.003125% to 4%) do not exhibit antimicrobial activity. However, various tested combinations of Cl at 0.0125%-0.05% with Ethyl Hexanol Ethoxylated Propoxylate at 0.0625% - 4% showed >3 log reduction in E. hirae counts indicating strong synergistic antimicrobial effect of Cl and Ecosurf™ EH-9. Unlike the observation with Decyl Glucoside in Example 3 above, higher concentrations of Ethyl Hexanol Ethoxylated Propoxylate did not affect the overall antimicrobial activity. The Cl to Ethyl Hexanol Ethoxylated Propoxylate ratios ranging from 0.003125:1 - 0.8:1 exhibits strong synergy in their antimicrobial efficacy that kills E. hirae effectively. Table 6A. Antimicrobial efficacy: log reduction achieved with Cl + Ecosurf™

[0140] EH-9 after 1 minute contact time at 20°C vs E. hirae

[0141] Example 4B: Antimicrobial efficacy of combination of Cl, Non ionic surfactant ( Decyl glucoside (DG) and Ecosurf™ EH-9 ) Further combination of AMP C 1 , Decyl glucoside (DG) and Ecosurf™ EH-9 (Ethyl

[0142] Hexanol Ethoxylated Propoxylate) were tested for activity on bacteria.

[0143] It was found that the combination in ratio of 0.0125:1:3 of Cl, Decyl glucoside (DG) and EH9 provided more than 5 log kill value in a minute when tested on various bacteria. The results are provided below in Table 6B Table 6B. Antimicrobial efficacy: log reduction achieved with Cl + Non-ionic surfactants (DG + Ecosurf™ EH9) after 1 minute contact time at 20°C

[0144] Example 5: Sanitizer and Disinfectant formulation

[0145] Formulations were developed to realize the utility of synergistic activity of AMP Cl with surfactants, Decyl glucoside or Ethyl Hexanol Ethoxylated Propoxylate (Ecosurf™ EH9). Strong bacterial killing activity makes them useful as Sanitizers (primarily used on living surfaces such as hand sanitizers) and disinfectants (used on non-living surfaces such as door knobs, kitchen surfaces, tables etc). The antimicrobial activity of these formulations was tested for rapid bactericidal activity as per EN 1276 standard protocol using four bacteria prescribed by the standard. Bacteria tested were Staphylococcus aureus ATCC 6538, Enterococcus hirae ATCC 10541, Escherichia coli ATCC 10536 & Pseudomonas aeruginosa ATCC 15442. The conditions used for the assay was dirty (0.3% final BSA concentration) and contact time was 1 minute. Formulation SI, S2 and S4 failed to demonstrate bacterial killing capability against all four bacteria. The synergistic combinations of Cl with either Decyl glucoside or Ethyl Hexanol Ethoxylated Propoxylate in prepared formulation S3 and S5 exhibited >3 log reduction of viable cell counts against all four tested bacteria after 1 minute contact time. Table 7. Alcohol free Sanitizer & Disinfectant formulation and its antimicrobial efficacy estimated as log reduction of viable counts of E. coli, S. aureus, P. aeruginosa and E. hirae achieved after 1 minute contact time at 20°C.

[0146] 5

[0147] Example 6: Sanitizer & Disinfectant formulation with Malic acid as chelator

[0148] Most formulations use chelator to take care of water hardness / metal ions and ensure that the actives in the formulation can work without interference from metal 10 ions. Chelators also protect against rancidity and color changes caused by heavy metal ions. EDTA was used as chelator to sequester di and trivalent metal ions (Formulation SI - S5). Further iterations demonstrated that 0.05% disodium EDTA is sufficient for bactericidal activity against four test bacteria as demonstrated by formulation S6 (Table 8). Malic acid was used in formulation S7 - S9. Use of Malic acid at 0.15% provided >5 log reduction activity against all four bacteria tested. Thus, Malic acid is suitable as the chelating agent for sanitizer and disinfectant 5 formulation.

[0149] Table 8. Sanitizer & Disinfectant formulation and its antimicrobial efficacy estimated as log reduction of viable counts of E. coli, S. aureus, P. aeruginosa and E. hirae achieved after 1 minute contact time at 20°C.

[0150] 10 Example 7: Antimicrobial Alcohol-free Deodorant

[0151] Deodorants are used to overcome the body malodour that develops over time primarily in underarm, foot and groin areas. The malodour is developed due to microbial metabolism on the surface of skin [Rudden, M., Herman, R., Rose, M. et al. The molecular basis of thioalcohol production in human body odour. Sci Rep 10, 12500 (2020). https: / / doi.org / 10.1038 / s41598-020-68860-z]. Examples of odorant molecules include volatile molecules such as 3-methyl-3-sulfanylhexan-l- ol (MSSH), 3-methyl-2-hexenoic acid (3M2H) & 3 -hydroxy-3 -methylhexanoic acid (HMHA) that are produced in underarm skin area from the sebaceous secretions of human apocrine glands by some members of Corynebacterium present on human skin, including Corynebacterium striatum. Short chain fatty acids such as isovaleric acid with characteristic cheesy odour are produced by Staphylococcus epidermidis present on human skin by metabolizing leucine secreted in sweat, particularly on feet [James, A.G., Cox, D. and Worrall, K., 2013. Microbiological and biochemical origins of human foot malodour. Flavour and Fragrance Journal, 28(4), pp.231-237, https: / / doi.org / 10.1002 / ffj.3136].

[0152] The body odour is controlled using either antiperspirants or deodorants. Antiperspirants reduces extent of sweating there by decreasing the food available for bacteria present on skin. The deodorant formulations works by either killing microbes through use of alcohols (ethanol) & other antimicrobials or / and by masking the malodour using fragrances. Most deodorants contain >50% ethanol and routine application causes skin dryness. Underarm skin also develops pigmentation (melanin production) resulting in darker underarms compared to other skin areas. Hence, it is recommended to use alcohol free deodorant

[0153] Spray formulations DI - D4 were prepared as per composition detailed in Table 9 for use as deodorant that can be applied on skin of various areas such as underarm, foot, back etc. The prepared formulations D2 & D4 containing synergistic antimicrobial combinations exhibited >4 log reduction against both S. epidermidis & C. striatum after 1 minute contact time (Table 9). Formulation DI and D3 that do not contain Cl exhibited limited log reduction activity (<1.2). The proposed deodorant spray formulation D2 and D4 can be applied in form of fine mist on skin to kill bacteria responsible for odour production present on underarm, back, foot and other areas.

[0154] Table 9. Alcohol free Deodorant formulation and its antimicrobial efficacy estimated as log reduction of viable counts of S, epidermidis and C. striatum achieved after 1 minute contact time at 20 C.

[0155] 10

[0156] Example 8: Antimicrobial alcohol-free wipes

[0157] The antimicrobial wipe formulations were tested for rapid bactericidal activity as per EN 1276 standard protocol using 4 bacteria prescribed by the standard. Bacteria tested were Staphylococcus aureus ATCC 6538, Enterococcus hirae ATCC 10541, Escherichia coli ATCC 10536 & Pseudomonas aeruginosa ATCC 15442. The conditions used for the assay was dirty (0.3% final BSA concentration) and contact time was 1 minute.

[0158] 5

[0159] Wipe formulation W1 - W4 were prepared incorporating fragrance solubilizer, humectant, chelator and preservative. The wet wipes base, Formulation Wl, showed significant activity against Gram negative bacteria E. coli and P. aeruginosa but has no bactericidal activity against S. aureus and E. hirae (Gram 10 positive bacteria). Thus, it is unsuitable for application as disinfectant wipes. The use of synergistic combination of Cl and surfactant (Decyl glucoside or Ethyl Hexanol Ethoxylated Propoxylate ) in prepared formulation W2, W3 and W4 provided >4.5 log reduction of viable cell counts of all four tested bacteria after 1 minute contact time.

[0160] 15

[0161] Table 10. Antimicrobial alcohol-free wipe formulation and its antimicrobial efficacy estimated as log reduction of viable counts of E. coli, S. aureus, P. aeruginosa and E. hirae achieved after 1 minute contact time at 20°C.

[0162] Table 11. Antimicrobial alcohol-free wipe formulation with Malic acid replacing EDTA and its antimicrobial efficacy estimated as log reduction of viable counts of E. coli, S. aureus, P. aeruginosa & E. hirae achieved after 1 minute contact time at 20 C.

[0163] 5

[0164] The present invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention, in addition to those described herein, will become apparent to those skilled in the art from the 5 foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims. The invention is, therefore, to be limited only by the terms of the appended claims along with the full scope of equivalents to which the claims are entitled.

Claims

CLAIMS:

1. An antimicrobial composition comprising an effective concentration of cathelicidin peptide fragment (FK16) as set forth in SEQ ID 1, or salts and derivatives thereof and an effective concentration of at least one non-ionic surfactant.

2. The antimicrobial composition as claimed in claim 1, wherein said antimicrobial composition achieves a log-kill of at least 3, after a contact time of about 1 min against at least one microbe3. The antimicrobial composition as claimed in claim 1 or claim 2, wherein the effective concentration of FK16 (Cl) peptide is 0.0125%-0.05% with respect to the total weight of the composition.

4. The antimicrobial composition as claimed in claim 1 or claim 2, wherein the at least one non-ionic surfactant is selected from Alkyl-Poly Glucosides (APG), Secondary alcohol ethoxylates, or combinations thereof.

5. The antimicrobial composition as claimed in claim 4, wherein the Alkyl- Poly Glucosides (APG) is at least one selected from the group consisting of Decyl-Glucoside, caprylyl / capryl glucoside.

6. The antimicrobial composition as claimed in claim 4, wherein the Secondary alcohol ethoxylates is Ethyl Hexanol Ethoxylated Propoxylate.

7. The antimicrobial composition as claimed in claim 4, wherein the non-ionic surfactant is Decyl-Glucoside and / or Ethyl Hexanol Ethoxylated Propoxylate.

8. The antimicrobial composition as claimed in claim 5, wherein Decyl- Glucoside is present at a concentration of 0.0625% to 0.5% with respect to the total weight of the composition.

9. The antimicrobial composition as claimed in claim 6, wherein Ethyl Hexanol Ethoxylated Propoxylate is present at a concentration of 0.0625% to 4% with respect to the total weight of the composition.

10. The antimicrobial composition as claimed in claim 7, wherein Decyl- Glucoside to Ethyl Hexanol Ethoxylated Propoxylate in a ratio ranging from 1.6: 1 to 2.4: 1.

11. The antimicrobial composition as claimed in claim 1 or 2, wherein the antimicrobial peptide and the non-ionic surfactant are combined at a ratio ranging from 0.003125: 1 to 0.8: 1.

12. The antimicrobial composition as claimed in claim 1 or 2, wherein the antimicrobial composition further comprises a chelating agent.

13. The antimicrobial composition as claimed in claim 12, wherein the chelating agent is disodium EDTA.

14. The antimicrobial composition as claimed in claim 12, wherein the chelating agent is selected from the group consisting of environment friendly and biodegradable ingredients like malic acid, citric acid, GLDA (Glutamic acid Di Acetate) and PESA (Polyepoxysuccinic acid).

15. The antimicrobial composition as claimed in claim 12, wherein disodium EDTA is present at a concentration of 0.1 to 0.4% with respect to the total weight of the composition.

16. The antimicrobial composition as claimed in claim 13, wherein the chelating agent is present at a concentration of 0.01 to 2% with respect to the total weight of the composition.

17. The antimicrobial composition as claimed in claim 2, wherein at least one microbe is selected from Gram negative bacteria Pseudomonas aeruginosa, Escherichia coli and Gram positive bacteria Staphylococcus aureus, Enterococcus hirae.

18. An antimicrobial composition comprising an effective concentration of cathelicidin peptide fragment (FK16) as set forth in SEQ ID 1, or salts and derivatives thereof, an effective concentration of at least one non-ionic surfactant selected from Decyl-Glucoside, Capryl glucoside, Caprylyl / Capryl Glucoside, Ethyl Hexanol Ethoxylated Propoxylate, or combinations thereof, and an effective concentration of at least one chelating agent selected from disodium EDTA, malic acid, citric acid, GLDA and PESA such that the antimicrobial composition achieves a logkill of at least 3, after a contact time of about 1 min against at least one microbe.

19. The antimicrobial composition as claimed in any one of the preceding claims, further comprising cosmetically acceptable excipients selected from, emulsifiers, solvents, rheology modifying agents, fillers / thickening agents, structurants, emollients, diluents, humectants, feel enhancers, preservatives and fragrances.

20. The antimicrobial composition as claimed in any one of the preceding claims, wherein the antimicrobial compositions is in a form selected from wipes, hand wash, deodorants, sanitizer, body wash, soap, shampoo and gels, wound healing cream / lotion and surface cleaner.

21. An alcohol-free wet wipe formulation comprising the antimicrobial composition as claimed in claim 1 or claim 17.

22. The alcohol-free wet wipe formulation as claimed in claim 20, further comprising at least one excipient selected from the group consisting of preservative, pH modifier, humectant, fragrance, and fragrance solubilizer.

23. A method of disinfecting a surface, the method comprising the steps of: providing an alcohol-free wet wipe as claimed in claim 20; and wiping the surface with the wet wipe.

24. An alcohol-free deodorant formulation comprising the antimicrobial composition as claimed in claim 1 or claim 17.

25. The alcohol-free deodorant formulation as claimed in claim 24 further comprising at least one excipient selected from the group consisting of preservative, fragrance, and fragrance solubilizer.

26. The alcohol-free deodorant formulation as claimed in claim 24, wherein the alcohol-free deodorant formulation is in the form of a mist spray format.

27. A method of reducing body odour, the method comprising the steps of: providing an alcohol-free deodorant as claimed in claim 25 and applying the deodorant to the skin in mist spray format.

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