WO2014120247A1 - Combinaisons de sophorolipides modifiés en tant qu'agents antimicrobiens - Google Patents

Combinaisons de sophorolipides modifiés en tant qu'agents antimicrobiens Download PDF

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WO2014120247A1
WO2014120247A1 PCT/US2013/024537 US2013024537W WO2014120247A1 WO 2014120247 A1 WO2014120247 A1 WO 2014120247A1 US 2013024537 W US2013024537 W US 2013024537W WO 2014120247 A1 WO2014120247 A1 WO 2014120247A1
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oil
admixture
natural
sophorolipid
agents
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PCT/US2013/024537
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English (en)
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Richard A. Gross
Thavasi Renga THAVASI
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Polytechnic Institute Of New York University
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Priority to PCT/US2013/024537 priority Critical patent/WO2014120247A1/fr
Priority to EP13874035.2A priority patent/EP2951311A4/fr
Publication of WO2014120247A1 publication Critical patent/WO2014120247A1/fr

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/90—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
    • A01N63/32—Yeast

Definitions

  • modified sophorolipid combinations as antimicrobial agents to control pathogens and other microbial strains that includes, but is not limited to, plant, animal and human pathogens, biofilm forming microbes, biofouling microbes, algae, fungi, bacteria, virus and protozoa.
  • Sophorolipids are glycolipid biosurfactant molecules produced by yeasts, such as Candida bombicola, Yarrowi alipolytica, Candida apicola, and Candida bogoriensis.
  • Microbial biosurfactants are surface active compounds produced by various microorganisms. They lower surface and interfacial tension and form spherical micelles at and above their critical micelle concentration (CMC).
  • CMC critical micelle concentration
  • Microbial biosurfactants generally have an amphiphilic structure, possessing a hydrophilic moiety, such as an amino acid, peptide, sugar or oligosaccharide, and a hydrophobic moiety including saturated or unsaturated lipid or fatty acids.
  • SLs consist of a hydrophilic carbohydrate head, sophorose, and a hydrophobic fatty acid tail with generally 16 or 18 carbon atoms with saturation or un- saturation.
  • Sophorose is an unusual disaccharide that consists of two glucose molecules linked ⁇ -1 ,2.
  • sophorose in SLs can be acetylated on the 6'- and/or 6"- positions (FIG. 1 ).
  • One fatty acid hydroxylated at the terminal or subterminal ( ⁇ -1 ) positions is ⁇ -glycosidically linked to the sophorose molecule.
  • the fatty acid carboxylic acid group is either free (acidic or open form) or internally esterified generally at the 4"-position (lactonic form) (FIG.
  • the hydroxy fatty acid component of SLs generally has 16 or 18 carbon atoms with generally one unsaturated bond (Asmer et al. 1988; Davila et al. 1993). However, the SL hydroxy fatty acid can also be fully saturated, di-unsaturated or tri-unsaturated. As such, SLs synthesized by C. bombicola consist of a mixture of related molecules. Differences between these molecules are found based on: i) the fatty acid structure (degree of unsaturation, chain length, and position of hydroxylation), whether they are produced in the lactonic or ring-opened form, and ii) the acetylation pattern.
  • lactonic and acidic SLs are synthesized in vivo from stearic acid with similar yields to oleic acid-derived SLs (Felse et al. 2007).
  • physiological variables during fermentations have provided routes to the variation of SL compositions.
  • sophorolipids of different structure As noted above, fermentation by different microorganisms, Candida bombicola, Yarrowi alipolytica, Candida apicola, and Candida bogoriensis, leads to sophorolipids of different structure noted above, the variations in sophorolipids based on fatty acid feedstocks and organisms leads to a wide array of sophorolipids including lactonic and acidic structures.
  • An additional modification that is relevant to acidic sophorolipids is cleavage of the sophorose moiety to the corresponding glucose-based glucolipids.
  • CMC critical micelle concentration
  • minimum surface tension have an inverse relationship with the alkyl ester chain length. That is, CMC decreased to 1/2 per additional CH 2 group for the methyl, ethyl, and propyl series of chain lengths.
  • Hydrogen bonding is proposed to be the primary driving force for adsorption of the sophorolipids on alumina.
  • Increase in the n-alkyl ester chain length of SLs caused a shift of the adsorption isotherms to lower concentrations.
  • the magnitude of the shift corresponds to the change in cmc of these surfactants.
  • sophorolipid natural mixture (non- chemically modified) is active against fungal plant pathogens Phytophthora sp. and Pythium sp. that are responsible for dumping-off disease. Inhibition of mycelial growth and zoospore motility was observed at high concentrations (200 mg/L and 50 mg/L, respectively). Thus, natural sophorolipids may be economically produced but have relative low activity against plant pathogen strains.
  • MSLs modified sophorolipids
  • MSLs have antibacterial, antiviral, and anti-inflammatory properties
  • MSLs were shown to down-regulate expression of pro-inflammatory cytokines including interleukin (Hagler et al. 2007).
  • the antibacterial activity of SLs can be increased by up to 1 ,000 times relative to the natural SL mixture by simple modifications such as esterification of fatty acid carboxyl groups and selective acetylation of disaccharide hydroxyl groups.
  • rhamnolipids have zoosporicidal activity against species of Pythium, Phytophora, and Plasmopara at concentrations ranging over 5 to 30 pg/nriL. Rhamnolipids are believed to intercalate with and disrupt plasma membranes. Indeed, US EPA Presidential Green Chemistry awards were recently presented to Agraquest Inc. and Jeneil Biosurfactant Companies for their work in developing rhamnolipid and lipopeptide biopesticide products, respectively. As mentioned above, rhamnolipids were found to be effective biopesticides and these results led to their commercialization by Jeneil Biosurfactants Co., LLC.
  • US Patent Publication US2005/0266036 describes rhamnolipids, a glycolipid biosurfactant produced by Pseudomonas aeruginosa, displays pesticidal activity on account of their cell wall penetration effect.
  • rhamnolipids a glycolipid biosurfactant produced by Pseudomonas aeruginosa
  • US Patent Publication 2012/0220464 A1 describes that addition of natural SLs and MSLs in pesticide formulation has increased the performance of the pesticide as a adjuvant.
  • the discovery herein is unique, distinct and could not be anticipated from US2012/0220464 A1 .
  • the present patent application describes how large enhancements in activity are achieved by combining an MSL with one or more natural SLs, or an MSL is combined with a different MSL.
  • the enhancements in antimicrobial activity observed herein are above what could be anticipated by one skilled in the art. See, also, WO 2003/043593 A1 .
  • the novel method described herein that is termed the "combination invention” is that by: i) mixing two MSLs or mixing more than two MSLs, ii) mixing one MSL or more than one MSL with a SL component of the natural mixture, or iii) mixing one MSL or more than one MSL with one or more SL components of the natural mixture results in synergistic effects whereby the combination of compounds results in much higher activity of the additive contributions of each of the components alone to reduce the proliferation of pathogenic bacteria, fungi, their spores and normal microbial strains.
  • Applications for this invention are broad and encompass the use of this "combination invention” as antimicrobials for environmental, industrial and medical fields.
  • the surprising and novel findings disclosed in this invention is that by: i) mixing two MSLs or mixing more than two MSLs, ii) mixing one MSL or more than one MSL with a SL component of the natural mixture, or iii) mixing one MSL or more than one MSL with one or more SL components of the natural mixture, the resulting antimicrobial activity is much greater than the additive effects of the components alone (the "combination invention”).
  • Applications of the combination invention include but are not limited to the following: i) to kill or inhibit the growth of pathogens and normal microbial strains such as pathogens of plants, animals and humans; ii) biofilm forming microbes; and iii) bio-fouling microbes.
  • Microbes that can be killed or inhibited by the "combination invention” include but are not limited to algae, fungi, bacteria, virus and protozoa.
  • MSL derivatives disclosed herein are described based on the predominant fatty acid constituent, 17-hydroxyoleic acid, produced by C. bombicola when fed crude oleic acid as its fatty acid source.
  • changes in the lipid feed canola oil and rapeseed oil
  • variations in feedstock also will result in changes in composition of MSL structures that are disclosed herein.
  • the new combination invention for use with natural SLs and MSLs disclosed herein provides for unique compositions relative to known natural SL and MSL derivatives in previous art that were discovered to be highly effective against commercially important plant pathogens, human pathogens, animal pathogens and undesired environmental microorganisms.
  • natural SLs may be economically produced but have relative low activity against pathogenic and normal microbes.
  • the present invention discloses a solution to this problem that involves the discovery of combination of natural SLs and MSLs that enhance their activities against pathogenic microbes and normal microbial strains.
  • X 1 or X 2 can be oxymethyl (-CH 2 O-) or methylene (-CH 2 -);
  • R 1 and/or R 2 can be selected from the following functional groups: hydrogen, acetyl, acryl, urethane, hydroxyalkyl, ether, halide, carboxyalkyl or alkyl containing heteroatoms ( , 2 ° , and 3 ° amino, tetraalkylammonium, sulfate, phosphate);
  • R 3 can be a hydrogen or alkyl group
  • the methods involved in performing these chemical transformations are well known to those skilled in the art;
  • X 3 can contain heteroatoms (e.g., O, S, NH); and
  • X 3 R 3 can be selected from the following functional groups: hydroxy, alkanethiolate, amide, alkanamide, alkanamide containing heteroatoms ( , 2 ° , and 3 ° amino, tetraalkylammonium), alkylsulfate, alkylphosphate, carbohydrate, mono- or oligopeptide with 2 - 50 amino acids.
  • FIG. 1 shows the structure of lactonic and open chain (acidic) forms of sophorolipid mixture produced by Candida bombicola.
  • FIG. 2 shows formulas for sophorolipids and sophorolipid analogs of the present invention.
  • FIG. 3 shows Sophorolipids in the lactonic form.
  • FIG. 4 shows Sophorolipids in the open chain (acidic) form.
  • FIG. 5 shows representative ester derivatives of the open chain form.
  • FIG. 6 shows amide and related derivatives of the open chain form.
  • FIG. 9 shows peptide derivatives of the open chain form.
  • FIG. 10 shows trans alkylidenation derivatives of lactonic and open chain SLs.
  • FIG. 1 1 shows electrophile derivatives at sophorose ring.
  • Scheme 1 shows a summary of chemo-enzymatic chemistry developed to prepare a library of sophorolipid analogs (see Azim et al. 2006, Singh et al., 2003, Bisht et al, 2000, Bisht et al., 1999)
  • Scheme 2 shows a synthesis of diamide derivatives from lactonic sophorolipid using transalkylidenation followed by amidation reactions.
  • Embodiments of this invention are based on the discovery that by: i) mixing two MSLs or mixing more than two MSLs; ii) mixing one MSL or more than one MSL with a SL component of the natural mixture; iii) mixing one MSL or more than one MSL with one or more SL components of the natural mixture, such mixtures result in synergistic effects whereby the combination of compounds results in much higher activity then the additive contributions of each of the components alone to kill or inhibit the growth of pathogens such as pathogens of plants, animals and humans as well as normal bacteria that grow on surfaces (e.g. bio-fouling microbes).
  • Microbes that can be killed or inhibited by the "combination invention” include but are not limited to algae, fungi, bacteria, virus and protozoa.
  • Embodiments of this invention include formulation of MSLs, natural SLs and their combinations with inert ingredients as listed in EPA's eligible inert ingredients list (see, e.g., http://www.epa.gov/opprd001/inerts/section25b_inerts.pdf) and any other material that could be used as an inert ingredient in the future.
  • MSL combinations described in this patent also include other MSL compositions that would be obvious to one skilled in the art based on review of this application or those encompassed within prior art.
  • Natural SLs and MSLs suitable for use in this invention include the following chemical compositions.
  • a second class of MSLs includes esterified ring-opened sophorolipids. Esterification of sophorolipids is achieved by alcoholysis of natural sophorolipid mixtures. Esters of varying chain lengths and with varying degrees of branching and containing a variety of heteroatoms are included in this invention (FIG. 5). Moreover, methods are already disclosed in the literature that describe selective acetylation of SLs at the 6'- and/or 6"-hydroxy sophorose groups. Therefore one skilled in the art will recognize that many variants may be generated by permutations of the ester functional group and sophorose acetyl groups.
  • a third class of sophorolipid derivatives includes amides of acidic sophorolipids.
  • sophorolipid amide derivatives are shown in FIG. 6.
  • sophorolipid amides can be synthesized from the sophorolipid methyl ester derivative 6 by treatment with an amine at elevated temperature. It is contemplated that a variety of amines, diamines, triamines of differing chain lengths containing aliphatic, olefinic, acetylenic, and aromatic substituents can be used to synthesize the corresponding amide derivatives.
  • amides derived from biogenic amines including, but not limited to, 4- aminosalicylic acid, 5-aminosalicylic acid, octopamine, 3-hydroxytyramine, phenethylamine, tryptamine, histamine, spermine, spermidine, 1 ,5-diaminopentane.
  • amides bearing at the sophorose head group ionic moieties such as sulfate, sulfonate, phosphate, carboxylate and quarternary ammonium salts that result in cationic or anionic charged head groups.
  • substituted amino-containing compounds can be used as a platform to expand the family of sophorolipid amides and that amino acids and polypeptides of varying chain lengths and composition can be incorporated (FIG. 9).
  • a fourth class of MSL includes ammonium salts derived from SL-amides with ⁇ ', ⁇ '-dimethylamino moieties.
  • An exemplary reaction is conversion of the sophorolipid ⁇ ', ⁇ '-dimethylethylamide derivative into the corresponding ammonium salt by treatment with methyl iodide at elevated temperature.
  • the quaternary ammonium salt may be prepared from alkyl halides of varying chain length as well as ⁇ , ⁇ , ⁇ -diiodoalkanes, leading to the formation of a wide array of sophorolipid structures.
  • a fifth class of MSLs include those modified at the sophorose 6' or 6" positions by, inter alia, an activated acyl molecule such as the vinyl ester or alkyl ester of propionic acid catalyzed by an enzyme catalyst such as a lipase in conjunction with one or more of the modifications described herein.
  • an activated acyl molecule such as the vinyl ester or alkyl ester of propionic acid catalyzed by an enzyme catalyst such as a lipase in conjunction with one or more of the modifications described herein.
  • an enzyme catalyst such as a lipase in conjunction with one or more of the modifications described herein.
  • the unsubstituted open-chain acidic sophorolipid is acetylated at the sophorose 6'-hydroxyl position.
  • carbonyl compounds of varying chain lengths and degrees of branching can be incorporated and that a variety of carbonyl-containing functional groups can be incorporated including succinate, malate and citrate.
  • esters of amino acids and oligopeptides can be incorporated at the 6' and/or 6" positions of the sophorose ring.
  • the 6' and/or 6" positions of the sophorose ring may be alkylated (FIG. 1 1 ) by ethylene oxide or a substituted alkylene oxide such as 2,3-epoxypropyl- 1 ,1 ,1 -trimethylammonium chloride (Quab151 ) or related electrophiles as described by Solarek (1989). Such substitutions will likely occur at the primary ( ) 6' and/or 6" positions but may also occur at the secondary (2°) sophorose ring hydroxy! groups to generate mixtures of sophorolipid derivatives.
  • combinations of metathesis (performed on either the lactonic or open chain SL) and chemical modification can be anticipated.
  • the cross metathesis of lactonic sophorolipid with vinyl acrylate will produce a diester wherein each of the ester groups can be converted into the corresponding amide derivative (Scheme 2).
  • Sophorolipid derivatives and sophorolipid components of the natural mixture used in bacterial and fungal plant pathogen assays.
  • the hydroxylated fatty acid of the natural mixture is predominantly 17-hydroxyoleic acid.
  • other fatty acid constituents with variations in chain length and unsaturation may also be present.
  • R 3 CHOHCH 2 (p-Phenol) 35
  • Example 1 Antifungal activity of MSL combinations against plant fungal pathogens.
  • Antifungal activity of MSL combinations were confirmed by experiment and observations.
  • the compounds used in antifungal assays are 1, 2, 6, 7, and 8.
  • MSLs and natural SLs single component or mixture
  • MSL + MSL or MSL + natural SL were tested individually (i.e., only one MSL) and in combinations (i.e., MSL + MSL or MSL + natural SL) against a panel of 18 different fungi.
  • mixtures consisted of equal quantities of each component of the mixture (i.e., 1 :1 ratio of MSL-X + MSL-Y, w/w).
  • the natural SL (single component or mixture) and MSL samples were dissolved in 5% (w/v) Tween 20 and 5% (w/v) Propylene glycol solution to a final concentration of 10 mg/mL (i.e., 5 mg of MSL-X and 5 mg of MSL-Y) that was used as a stock solution.
  • the stock solution (100 ⁇ ) was added into a 96 well microplate and serially diluted from 10 mg/mL to 0.0024 mg/mL using culture medium.
  • the culture media used for antifungal assay include, mineral salts medium (for Botrytis cinerea), corn meal broth (for Phytopthora infestans and P. capsici) and potato dextrose broth for all other fungi.
  • Botrytis cinerea 0.009 0.009 10 5 2.5 10 1 .25 10 -
  • Ustilago maydis - - - - - 1 .25 2.5 10 -
  • F. cereal is 2.5 - 5 1 .25 10 - 10 - -
  • Example 2 Antibacterial activity of MSL combinations against plant bacterial pathogens.
  • Bacterial infections in plants are much like the symptoms in fungal plant diseases. Examples are leaf spots, blights, wilts, scabs, cankers and soft rots of roots, storage organs and fruit, and overgrowth.
  • 7 different plant pathogenic bacteria were used (Table 4). The compounds used in antibacterial assay are 1 , 2, 6, 7, 8, 16. In this assay, MSLs were tested individually (i.e., only one MSL) and in combinations (i.e., MSL + MSL or MSL + natural SL) against a panel of 7 different bacateria.
  • mixtures consisted of equal quantities of each component of the mixture (i.e. ,1 :1 ratio of MSL-X + MSL-Y, w/w). Combinations were dissolved in 5% (w/v) Tween 20 and 5% (w/v) propylene glycol solution to a final concentration of 10 mg/mL that was used as a stock solution.
  • the stock solution 100 ⁇ was added into a 96 well microplate and serially diluted from 10 mg/mL to 0.0024 mg/mL using culture medium.
  • the culture media used for antibacterial assay is Tryptic Soy Broth.
  • Example 3 Antibacterial activity of MSL combinations against biofilm forming bacterial strains.
  • MSL samples were dissolved in 5% (w/v) Tween 20 and 5% (w/v) Propylene glycol solution to a final concentration of 10 mg/nnL that was used as a stock solution.
  • the stock solution (100 ⁇ ) was added into a 96 well microplate and serially diluted from 10 mg/nnL to 0.0024 img/mL using culture medium.
  • the culture media used for antibacterial assay is Tryptic Soy Broth. After serial dilution, 95 ⁇ of fresh culture medium and 5 ⁇ of bacterial cell suspension were added to each well and the plates were incubated for 24 to 48 h at 30 °C.
  • Antibacterial activity was determined by measuring the optical density (OD) of micro wells containing MSL and bacterial culture at 540 nm in a spectrophotometer. A control was maintained for each bacterial culture without adding MSL into the culture medium. The difference in OD between MSL added wells and the control was calculated and converted into %-growth inhibition. The formula used for the calculation of %-growth inhibition is: [Control OD - OD of MSL added wells/Control OD] x 100. The MSLs and natural SL tested in this assay showed 55 to 98 % growth inhibition activity against the 8 bacterial strains tested. The results are shown in Table 6. Table 6. Antimicrobial activity of natural SLs and MSLs against biofouling bacterial strains
  • This invention demonstrates that by: i) mixing two MSLs or mixing more than two MSLs; ii) mixing one MSL or more than one MSL with a SL component of the natural mixture; or iii) mixing one MSL or more than one MSL with one or more SL components of the natural mixture, such mixtures result in synergistic effects whereby the combination of compounds results in much higher activity then the additive contributions of each of the components alone to kill or inhibit the growth of pathogens such as pathogens of plants, animals and humans as well as normal bacteria that grow on surfaces (e.g. bio-fouling microbes).
  • Microbes that can be killed or inhibited by the combination invention include but are not limited to algae, fungi, bacteria, virus and protozoa.
  • This invention demonstrates that by: i) mixing two MSLs or mixing more than two MSLs; ii) mixing one MSL or more than one MSL with a SL component of the natural mixture; or iii) mixing one MSL or more than one MSL with one or more SL components of the natural mixture, such mixtures result in synergistic effects whereby the combination of compounds results in much higher activity of the additive contributions of each of the components alone to kill or inhibit the growth of pathogens such as pathogens of plants, animals and humans as well as normal bacteria grow on surfaces (e.g. bio-fouling microbes).
  • Microbes that can be killed or inhibited by the combination invention include but are not limited to algae, fungi, bacteria, virus and protozoa.
  • This invention demonstrates that by: i) mixing two MSLs or mixing more than two MSLs; ii) mixing one MSL or more than one MSL with a SL component of the natural mixture; or iii) mixing one MSL or more than one MSL with one or more SL components of the natural mixture, such mixtures result in synergistic effects whereby the combination of compounds results in much higher antimicrobial activity then the additive contributions of each of the components alone.
  • MSLs incorporated in this invention also include those that result by performing multiple modification chemistries on a natural sophorolipid precursor.
  • a small subset of the large number of permutations that result from performing multiple modifications on a natural SL precursor are given in the following examples: i) hydrogenation of the fatty acid carbon-carbon double bond and esterification of the fatty acid carboxyl group; ii) hydrogenation of the carbon-carbon double bond and amidation at the carboxylate group; and iii) transalkylidination of the SL-lipid carbon-carbon double bond by reaction with methyl acrylate followed by amidation at the formed methyl ester moiety, vi) amidation at the carboxylate and acetylation at the sophorose head group.
  • the invention is a method for controlling microbes; inhibition of their growth and killing of live cells and spores, comprising: providing an admixture of compounds that consist of two or more constituents that include a natural sophorolipid component, natural sophorolipid mixture or a modified sophorolipid, and applying said admixture to a plant, surface, device, or any system containing microbes that have grown or might grow which are undesirable.
  • a natural sophorolipid component natural sophorolipid mixture or a modified sophorolipid
  • the various microbes to which the invention can be applied include known or not yet discovered plant pathogens; human pathogens; live cells or spores in the group that consists of: bacteria, fungi, viruses, algae and protozoan; microbes or combination of microbes that grow on surfaces causing fouling or contamination of that surface; and instances where a microbe has accumulated in access due to some shift in the ecosystem such as accumulation of a non-natural chemical in lakes.
  • microbes to which the invention can be applied include those that form biofilms that contaminate surfaces such as, but not limited to catheters, medical dives, walls, shower curtains, swimming pools, pipelines, water filters, cooling towers, marine structures, ships, boats, navigational aids, channel markers, buoys, and oil exploration plat forms.
  • Representative MSL derivatives can be synthesized by methods that are known from the prior art using natural sophorolipids produced by fermentation from a feedstock mixture, wherein the fatty acid is selected from the group consisting of tallow, sunflower oil, rapeseed oil, safflower oil, soya bean oil, palm oil, coconut oil, olive oil, and short-chain to medium chain length carboxylic acid having an alkyl chain length from 6 to 22 carbons.
  • the MSL derivatives can be obtained without purifying the reaction mixture or pure compounds of the same.
  • the MSL derivatives can be obtained from sophorolipid mixtures of different purity with varying contents of natural to open chain sophorolipids.
  • the admixture can consist of 2 MSL compounds of different compositions.
  • the admixture can consist of mixing 2 or more MSL compounds of different compositions.
  • the admixture can consist of mixing 1 MSL and 1 or more natural sophorolipid components.
  • the admixture can consist of mixing 1 MSL and a natural SL mixture.
  • the admixture can consist of mixing one or more MSLs and one or more natural sophorolipid components.
  • the admixture can be applied as a solution that can be a concentrate or at an appropriate concentration for use.
  • the admixture can be in powder form and applied as a powder or dissolved in a solution prior to application.
  • the compound mixture of the present invention acts synergistically to increase the antimicrobial activity relative to any of the components in the mixture tested alone.
  • the admixture acts synergistically in a ratio where the component with the lowest concentration is 1 :200 th (w/w) of the summation of the other components and the component of the admixture that is in the highest concentration is up to 30 times greater than the summation of the other components.
  • the admixture can further include chemical or biobased emulsifiers, biosurfactants, surfactants, and eco-friendly organic solvents used in pesticides, antimicrobial agent(s), disinfectant(s), personnel hygienic agents and cosmetics.
  • the admixture further include inert components used in the formulation of pesticides, biopesticides, biochemical pesticides and antimicrobial agents, disinfectants, personnel hygienic agents and cosmetics such as adjuvants, buffering agents or pH adjusting agents/salts and solublizers.
  • the physical form of formulated mixtures can be as a wettable powder, powders, dust, granules, liquids, gels, semisolids, colloidal materials, paste, incorporated in wipes, papers, polymers and in any other form a potential pesticide, biopesticide, biochemical pesticide, antimicrobial agent, disinfectant, personnel hygienic agent and cosmetics can be formulated.
  • Inert components suitable for use as an adjuvant or that may possess pesticide, antimicrobial, disinfectant, personnel hygienic activities include those of natural origin that complement the natural aspects of the natural SL, MSL derivative and MSL combinations and can be either an oil component such as cinnamon oil, clove oil, cottonseed oil, garlic oil, or rosemary oil; another natural biosurfactant or synthetic surfactant; or the component may be an aldehyde such as cinnamic aldehyde, ands wherein other oils that may be used as a pesticidal and antimicrobial component or adjuvants include: almond oil, camphor oil, castor oil, cedar oil, citronella oil, citrus oil, coconut oil, corn oil, eucalyptus oil, fish oil, geranium oil, lecithin, lemon grass oil, linseed oil, mineral oil, mint or peppermint oil, olive oil, pine oil, rapeseed oil, safflower oil, sage oils, sesam
  • Suitable buffering agents include organic and amino acids or their salts, wherein suitable buffers include citrate, gluconate, tartrate, malate, acetate, lactate, oxalate, aspartate, malonate, glucoheptonate, pyruvate, galactarate, glucarate, tartronate, glutamate, glycine, lysine, glutamine, methionine, cysteine, arginine and a mixture thereof, phosphoric and phosphorous acids or their salts, and wherein synthetic buffers are suitable to be used but it is preferable to use natural buffers such as organic and amino acids or their salts listed above.
  • Solubility control agents or excipients also can be used in the inventive formulations to control the release of the active substances, examples of which include wax, chitin, chitosan, C12-C20 fatty acids such as myristic acid, stearic acid, palmitic acid; C12-C20 alcohols such as lauryl alcohol, cetyl alcohol, myristyl alcohol, and stearyl alcohol; amphiphilic esters of fatty acids with glycerol, especially monoesters C12-C20 fatty acids such as glyceryl monolaurate, glyceryl monopalmitate; glycol esters of fatty acids such as polyethylene glycol monostearate or polypropylenemonopalmitate glycols; C12-C20 amines such lauryl amine, myristyl amine, stearyl amine, and amides of C12-C20 fatty acids.
  • Suitable pH adjusting agents include potassium hydroxide, ammonium hydroxide, ammoni
  • Additional components can be included in aqueous preparation formulations of thei inventive compositions, such as the salt form of polyprotic acids, examples of which include sodium bicarbonate, sodium carbonate, sodium sulfate, sodium phosphate, sodium biphosphate.
  • Suitable synthetic surfactant include alkyi betaines, alkyi sulfates, alkyi ammonium bromide derivatives, alkyi phenol ethoxylates, alkyi ethylene or polyethylene ethoxylates, alkyi or acyl glycosides, tween 80, tween 60, tween 40, tween 20, polypropylene glycol and biosurfactants.
  • Suitable biosurfactants or surface active compounds that are embodied in the combination invention can be formulated with one or more members of the consisting of other natural glycolipids that include rhamnolipids, mannosylerythritol, cellobiose lipids, trehalose lipids, emulsan, lipopeptides, surfactin, lipoproteins, lipopolysaccharide- protein complexes, phospholipids, and polysaccharide-protein-fatty acid complexes and any other compound(s) with potential uses as a biosurfactant.
  • other natural glycolipids that include rhamnolipids, mannosylerythritol, cellobiose lipids, trehalose lipids, emulsan, lipopeptides, surfactin, lipoproteins, lipopolysaccharide- protein complexes, phospholipids, and polysaccharide-protein-fatty acid complexes and any other compound(s)
  • biosurfactants or surface active compounds can be pure, crude or directly collected from culture broth or the culture broth having surface active agents in it.
  • the natural SL, MSL derivative and combinations thereof encompassed by the combination invention can be applied by spraying, pouring, dipping, in the form of concentrated or diluted liquids, solutions, suspensions, powders, incorporated in wipes, papers and polymers and the like, containing such concentrations of the active agent as is most suited for a particular purpose and application.
  • the natural SL, MSL derivative and combinations thereof encompassed by the combination invention can be formulated such that they are solid formulations that can different forms and shapes such as cylinders, rods, blocks, capsules, tablets, pills, pellets, strips, and spikes, and wherein solid formulations may also be milled, granulated or powdered, and wherein the granulated or powdered material may be pressed into tablets or used to fill pre-manufactured gelatin capsules or shells, ad wherein semi solid formulations can be prepared in paste, wax, gel, or cream preparations.
  • the natural SL, MSL derivative and combinations thereof encompassed by the combination invention can be used for human or animal applications; the formulations may be prepared in liquid, paste, ointment, suppository, capsule or tablet forms and used in a way similar to drugs used in the medicinal drugs industry, the formulations can be encapsulated using components known in the pharmaceutical industry so as to protect the components from undesirable reactions and help the ingredients resist adverse conditions in the environment or the treated object or body e.g. stomach.
  • the natural SL, MSL derivative and combinations thereof encompassed by the combination invention can be applied to plants, pests, or soil using various methods of application depending on certain circumstances.
  • the natural SL, MSL derivative and combinations thereof encompassed by the combination invention can be introduced directly in the soil in the vicinity of plant roots, in the form of liquid, bait, powder, dusting, or granules, or alternatively, the biopesticidal compositions may be inserted in the soil as tablets, spikes, rods, or other shaped moldings.
  • the natural SL, MSL derivative and combinations thereof encompassed by the combination invention can be formulated and used for treating individual plant, tree, plants or trees, for example, the formulations can be molded in different shapes or forms (solid, paste or gel, or liquid) and introduced into the vascular tissue of the plants, and wherein the moldings forms can be as tablets, capsules, plugs, rods, spikes, films, strips, nails, or plates, and wherein the shaped moldings can be introduced into pre- drilled holes into the plants or root flares, or they can be pushed or punched into the cambium layer.
  • the natural SL, MSL derivative and combinations thereof encompassed by the combination invention after formulation can be applied by the use of dispensing devices such as syringes, pumps or caulk guns, paste-tubes or plunger tubes for delivering semi-solid formulations (paste, gel, cream) into drilled holes in tree trunks or root flares.
  • dispensing devices such as syringes, pumps or caulk guns, paste-tubes or plunger tubes for delivering semi-solid formulations (paste, gel, cream) into drilled holes in tree trunks or root flares.
  • the natural SL, MSL derivative and combinations thereof encompassed by the combination invention after formulation can be applied in the form of paste, gel, coatings, strips, or plasters onto the surface of the plant, a plaster or strip may be in a semi-solid formulation, e.g., insecticide placed on the side that will contact the tree, bush, or rose during the treatment, and wherein the same strip may have glue or adhesive at one or both ends to wrap around or stick to the subject being treated.
  • a plaster or strip may be in a semi-solid formulation, e.g., insecticide placed on the side that will contact the tree, bush, or rose during the treatment, and wherein the same strip may have glue or adhesive at one or both ends to wrap around or stick to the subject being treated.
  • the natural SL, MSL derivative and combinations thereof encompassed by the combination invention after formulation can be sprayed or dusted on the leaves in the form of pellets, spray solution, granules, or dust.
  • the natural SL, MSL derivative and combinations thereof encompassed by the combination invention after formulation can be solid or semi-solid compositions that can be coated using film-coating compounds used in the pharmaceutical industry such as polyethylene glycol, gelatin, sorbitol, gum, sugar or polyvinyl alcohol, which is particularly essential for tablets or capsules used in pesticide formulations, and wherein film coating can protect the handler from coming in direct contact with the active ingredient in the formulations, and where, in addition, a bittering agent such as denatonium benzoate or quassin may also be incorporated in the pesticidal formulations, the coating or both.
  • film-coating compounds used in the pharmaceutical industry such as polyethylene glycol, gelatin, sorbitol, gum, sugar or polyvinyl alcohol, which is particularly essential for tablets or capsules used in pesticide formulations, and wherein film coating can protect the handler from coming in direct contact with the active ingredient in the formulations, and where, in addition, a bittering agent such as denatonium benzoate or quassi
  • compositions and application rate of the compositions may be varied widely depending on the pest, plant, animal, human, microbes or area treated, or method of application, wherein the compositions and methods of the invention can be used to control a variety of pests, microbes, including insects and other invertebrates, algae, and, in some situations, weeds or other plants.

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  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Dentistry (AREA)
  • Plant Pathology (AREA)
  • Pest Control & Pesticides (AREA)
  • Environmental Sciences (AREA)
  • Agronomy & Crop Science (AREA)
  • Mycology (AREA)
  • Biotechnology (AREA)
  • Virology (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

L'invention concerne un procédé pour lutter contre, inhiber et tuer des pathogènes et des souches microbiennes normales qui comprennent, mais sans y être limités, des pathogènes de plantes, d'animaux et d'êtres humains, des microbes formant un biofilm, des microbes de bio-encrassement, des algues, des champignons, des bactéries, des virus et des protozoaires à l'aide de dérivés naturels de SL, MSL et des combinaisons de ceux-ci, englobés par l'invention de combinaison.
PCT/US2013/024537 2013-02-02 2013-02-02 Combinaisons de sophorolipides modifiés en tant qu'agents antimicrobiens WO2014120247A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
PCT/US2013/024537 WO2014120247A1 (fr) 2013-02-02 2013-02-02 Combinaisons de sophorolipides modifiés en tant qu'agents antimicrobiens
EP13874035.2A EP2951311A4 (fr) 2013-02-02 2013-02-02 Combinaisons de sophorolipides modifiés en tant qu'agents antimicrobiens

Applications Claiming Priority (1)

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PCT/US2013/024537 WO2014120247A1 (fr) 2013-02-02 2013-02-02 Combinaisons de sophorolipides modifiés en tant qu'agents antimicrobiens

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WO2020190699A1 (fr) * 2019-03-15 2020-09-24 Locus Ip Company, Llc Matériaux et procédés pour le traitement et la prévention améliorés de biofilms
CN112442098A (zh) * 2019-09-03 2021-03-05 中国石油化工股份有限公司 一种磺化改性槐糖脂或其盐以及它们的制备方法
CN112742855A (zh) * 2019-10-31 2021-05-04 中国石油化工股份有限公司 一种石油污染后的土壤修复方法
US11045589B2 (en) 2017-09-22 2021-06-29 Becton, Dickinson And Company 4% trisodium citrate solution for use as a catheter lock solution
US11447430B2 (en) 2018-05-08 2022-09-20 Locus Agriculture Ip Company, Llc Microbe-based products for enhancing plant root and immune health
US11590231B2 (en) 2017-07-27 2023-02-28 Locus Solutions Ipco, Llc Compositions for enhancing bioavailability of pharmaceuticals, supplements and ingested substances
US11759544B2 (en) 2018-05-25 2023-09-19 Locus Solutions Ipco, Llc Therapeutic compositions for enhanced healing of wounds and scars
CN116199540B (zh) * 2023-01-28 2024-03-22 西北农林科技大学 一种提高作物耐盐性的氨基酸-鼠李糖脂混合液及其制备方法和应用
US11964040B2 (en) 2017-12-28 2024-04-23 Locus Solutions Ipco, Llc Oral health composition comprising purified biosurfactants and/or their derivatives
US11963528B2 (en) 2018-02-26 2024-04-23 Locus Solutions Ipco, Llc Materials and methods for control of insect pests using entomopathogenic fungi

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Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11590231B2 (en) 2017-07-27 2023-02-28 Locus Solutions Ipco, Llc Compositions for enhancing bioavailability of pharmaceuticals, supplements and ingested substances
US11045589B2 (en) 2017-09-22 2021-06-29 Becton, Dickinson And Company 4% trisodium citrate solution for use as a catheter lock solution
US11964040B2 (en) 2017-12-28 2024-04-23 Locus Solutions Ipco, Llc Oral health composition comprising purified biosurfactants and/or their derivatives
US11963528B2 (en) 2018-02-26 2024-04-23 Locus Solutions Ipco, Llc Materials and methods for control of insect pests using entomopathogenic fungi
US11447430B2 (en) 2018-05-08 2022-09-20 Locus Agriculture Ip Company, Llc Microbe-based products for enhancing plant root and immune health
US11759544B2 (en) 2018-05-25 2023-09-19 Locus Solutions Ipco, Llc Therapeutic compositions for enhanced healing of wounds and scars
WO2020190699A1 (fr) * 2019-03-15 2020-09-24 Locus Ip Company, Llc Matériaux et procédés pour le traitement et la prévention améliorés de biofilms
CN112442098A (zh) * 2019-09-03 2021-03-05 中国石油化工股份有限公司 一种磺化改性槐糖脂或其盐以及它们的制备方法
CN112442098B (zh) * 2019-09-03 2022-05-17 中国石油化工股份有限公司 一种磺化改性槐糖脂或其盐以及它们的制备方法
CN112742855A (zh) * 2019-10-31 2021-05-04 中国石油化工股份有限公司 一种石油污染后的土壤修复方法
CN116199540B (zh) * 2023-01-28 2024-03-22 西北农林科技大学 一种提高作物耐盐性的氨基酸-鼠李糖脂混合液及其制备方法和应用

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EP2951311A4 (fr) 2016-07-13
EP2951311A1 (fr) 2015-12-09

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