Antimicrobial surfactants and water borne coatings comprising the same

Abstract

The invention relates to novel antimicrobial surfactants and their application in antimicrobial coating systems, in particular water borne coatings. Provided is a method for providing an antimicrobial surfactant, comprising the steps of: (a) providing a hyperbranched polyurea having blocked isocyanates at the end of the polymer branches by the polycondensation of AB.sub.2 monomers; (b) introducing tertiary amine groups by reacting said blocked isocyanates of the hyperbranched polyurea with a tertiary amine compound that is functionalized with —OH, —NH.sub.2, —SH, or —COO—; and (c) quaternization of said tertiary amine groups by reacting with an alkylating agent to obtain a quaternized hyperbranched polymer having antimicrobial surfactant properties.

Claims

1. A method for providing an antimicrobial surfactant, comprising the steps of: (a) providing a hyperbranched polyurea having blocked isocyanates at the end of the polymer branches by the polycondensation of AB.sub.2 monomers, the AB.sub.2 monomers having the general formula I ##STR00010## wherein R.sub.1 and R.sub.2 are, respectively, aliphatic chains (CH.sub.2).sub.m and (CH.sub.2).sub.n wherein m and n are an integer in the range of 3 to 15, and wherein X is H, an aliphatic moiety or a polymerizable unit; (b) introducing tertiary amine (t-amine) groups by reacting said blocked isocyanates of the hyperbranched polyurea with a functionalized t-amine compound of the general formula
Y—(CH.sub.2).sub.n—NR.sup.1R.sup.2 wherein Y is OH or —NH.sub.2, n=2-12; R.sup.1 and R.sup.2 are independently selected from C.sub.1-C.sub.2 alkyl; and (c) quaternization of said t-amine groups by reacting with an alkylating agent to obtain a quaternized hyperbranched polymer having antimicrobial surfactant properties wherein said alkylating agent comprises a linear alkyl moiety of 8 to 12 carbon atoms.

2. An antimicrobial surfactant obtainable by a method comprising the steps of: (a) providing a hyperbranched polyurea having polymer branches with ends, and having blocked isocyanates at the ends of the polymer branches, the hyperbranched polyurea synthesized by polycondensation of AB.sub.2 monomers, the AB.sub.2 monomers having a structure of the general formula I ##STR00011## wherein R.sub.1 and R.sub.2 are, respectively, aliphatic chains (CH.sub.2).sub.m and (CH.sub.2).sub.n wherein m and n are integers in the range of 3 to 15, and wherein X is H, an aliphatic moiety or a polymerizable unit; (b) introducing tertiary amine (t-amine) groups by reacting said blocked isocyanates of the hyperbranched polyurea with a functionalized t-amine compound of the general formula
Y—(CH.sub.2).sub.n—NR.sup.1R.sup.2 wherein Y is —OH or —NH.sub.2; n=2-12; R.sup.1 and R.sup.2 are independently selected from C.sub.1-C.sub.2 alkyl; and (c) quaternization of said t-amine groups by reacting with an alkylating agent to obtain a quaternized hyperbranched polymer having antimicrobial surfactant properties, wherein said alkylating agent comprises a linear alkyl moiety of 8 to 12 carbon atoms.

3. Antimicrobial surfactant having a structure of the general formula II ##STR00012## wherein X is H, aliphatic or cycloaliphatic chain or an unsaturated moiety; n is an integer in the range of 3-15; z is an integer in the range of 2-12; Y is O or NH.sub.2; P.sub.1, P.sub.2 and P.sub.3 are linear alkyl chains, wherein one of said P.sub.1, P.sub.2 and P.sub.3 has a chain length of C.sub.8-C.sub.12 and the other two of P.sub.1, P.sub.2 and P.sub.3 have a chain length of C.sub.1-C.sub.2; p is an integer in the range of 1-25; and B is Br.sup.−, Cl.sup.−, I.sup.− or SO.sub.4.sup.2−, wherein the antimicrobial surfactant has a critical micelle concentration from 0.1 to 3.7 mg/mL.

4. Antimicrobial surfactant according to claim 3, wherein Y═O and z=3; or wherein Y═NH and z=2.

5. A composition comprising at least one antimicrobial surfactant according to claim 2.

6. Composition according to claim 5, wherein said composition further comprises a polymer produced from vinyl monomers, preferably styrenes, acrylates and/or methacrylates.

7. Composition according to claim 6, wherein said at least one antimicrobial surfactant is covalently incorporated in said polymer.

8. Composition according to claim 5, wherein said composition is an antimicrobial paint, an antimicrobial (two-component) coating composition, an antimicrobial impregnate, an antimicrobial adhesive, an antimicrobial sealant, an antimicrobial elastomer, an antimicrobial plastic, or an antimicrobial composite material.

9. A surface provided with an antimicrobial composition according to claim 5, preferably wherein said surface is a wall or a floor, more preferably a wall or a floor in a hospital, health care, day care or senior care building.

10. Antimicrobial surfactant according to claim 2, wherein X in step (a) is H.

11. Antimicrobial surfactant according to claim 2, wherein X in step (a) is selected from the group consisting of optionally substituted styrene, acrylate, methacrylate, vinylethers and fatty acids.

12. Antimicrobial surfactant according to claim 2, wherein said functionalized t-amine compound is N,N-dimethylethene diamine (DMEN), N,N-dimethylpropylene diamine (DMPN), 3-(dimethylamino)-1-propanol (DAMP), or N,N,N′-trimethyl-1,3-propane diamine.

13. Antimicrobial surfactant according to claim 2, wherein said alkylating agent in step (c) is an alkyl halide.

14. Antimicrobial surfactant according to claim 2, wherein steps (a), (b) and (c) are performed as a one-pot procedure.

15. Composition according to claim 5, wherein the composition is a water-born coating composition, a fabric additive, a clothing additive, a paint additive, an antimicrobial paint, an antimicrobial coating, an antimicrobial two-component coating composition, an antimicrobial impregnate, an antimicrobial adhesive, an antimicrobial sealant, an antimicrobial elastomer, an antimicrobial plastic, or an antimicrobial composite material.

16. Antimicrobial surfactant according to claim 3, wherein one of said P.sub.1, P.sub.2 and P.sub.3 has a chain length of C.sub.10-C.sub.12 and the other two of P.sub.1, P.sub.2 and P.sub.3 have a chain length of C.sub.1.

Description

LEGEND TO THE FIGURES

(1) FIG. 1: Exemplary antimicrobial hyperbranched surfactants of the invention. Panel A: compounds (herein referred to as “HBP—NH2-Cx” wherein x denotes the alkyl chain length) obtained using an amine-functionalized tertiary amine. Panel B: compounds (herein referred to as “HBP—OH-Cx” wherein x denotes the alkyl chain length) obtained using a hydroxyl-functionalized tertiary amine. Shown are variants quaternized with alkyl groups with various chain lengths (R═C1, C2, C4, C6, C8, C10 or C12).

(2) FIG. 2. .sup.1H NMR spectra of exemplary surfactants HBP—NH2-Cx, wherein x is 2, 4, 6, 8, 10 or 12.

(3) FIG. 3. .sup.1H NMR spectra of exemplary surfactants HBP—OH-Cx, wherein x is 1, 2, 4, 6, 8, 10 or 12.

(4) FIG. 4. Fluorescence emission spectra of Nile Red in HBP—OH—C2/water solution at varying concentrations (mg/mL).

(5) FIG. 5. Plot of maximum emission intensity of Nile Red versus the log of concentration (mg/mL) of (panel A) HBP—NH.sub.2—C2/water solution or (panel B) HBP—OH—C2/water solution—

EXPERIMENTAL SECTION

(6) Materials

(7) Carbonyl biscaprolactam (CBC, >99%) was kindly obtained from DSM Innovation Center. Bis (hexamethylene) triamine (BHTA, high purify), N,N-dimethylethenediamine (≥DMEN, ≥98.0%), 3-(dimethylamino)-1-propanol (DMAP, 99%), Tin(II)2-ethylhexanoate (92.5-100.0%), bromoethane (98%), 1-bromobutane (99%), 1-bromohexane (98%), 1-bromooctane (99%), 1-bromodecane (98%), and 1-bromododecane (97%), Nile Red, toluene and DMF were purchased from Sigma-Aldrich. All of the chemicals were used as received and without purification.

Example 1: Synthesis of ABs Monomers

(8) To a three-necked flask equipped with a reflux condenser, a nitrogen inlet and a connector to a vacuum pump, bis-hexamethylene triamine (BHTA, 49.5 g, 0.23 mol) and carbonyl biscaprolactam (CBC, 115.9 g, 0.46 mol) were added. After three cycles evacuating and flushing with nitrogen to remove the oxygen, the mixture was dissolved in 50 ml toluene and stirred at 80° C. overnight under nitrogen atmosphere. After the solution was cooled down to room temperature, toluene was removed under reduced pressure. Then the mixture was dissolved in 50 ml chloroform (CHCl.sub.3), and washed with saturated aqueous sodium chloride solution (8×500 ml) to remove impurities. The organic layer was dried with sodium sulfate, filtered to remove the salt. The solvent was removed under reduced pressure, yielding a white solid.

(9) .sup.1H-NMR (400 MHz, CDCl.sub.3): δ=1.34 to 1.56 (16H, m, CH.sub.2), 1.73 (12H, m, CH.sub.2 ring), 2.57 (4H, m, CH.sub.2NHCH.sub.2), 2.68 (4H, t, CH.sub.2CON), 3.26 (4H, m, CH.sub.2NH CO), 3.96 (4H, t, CH.sub.2NCO), 9.23 (2H, t, NHCO).

Example 2: Synthesis of Hyperbranched Polymers (HBP)

(10) To a three-necked flask equipped with a reflux condenser, a nitrogen inlet and a connector to a vacuum pump, AB.sub.2 monomers (8 g, 18.3 mmol) were added. After three cycles evacuating and flushing with nitrogen to remove the oxygen, the monomers were dissolved in 30 ml DMF and stirred at 145° C. 1 h under nitrogen atmosphere. After the solution was cooled down to room temperature, most of the DMF was removed under reduced pressure. Then the mixture was dissolved in 20 ml CHCl.sub.3 and washed several times with saturated aqueous sodium chloride solution to remove the impurities and residual DMF. The organic layer was collected and dried with sodium sulfate. The salt was removed by filtration the suspension and the solvent was removed under reduced pressure. Transparent yellow resin was obtained.

(11) .sup.1H-NMR (400 MHz, DMSO): δ=1.34 to 1.56 (m, CH.sub.2), 1.73 (m, CH.sub.2 ring), 2.57 (m, CH.sub.2NHCH.sub.2), 2.68 (t, CH.sub.2CON), 2.95 (t, CH.sub.2NHCO), 3.05 (t, CONCH.sub.2), 3.26 (m, CH.sub.2NH CO-end group), 3.96 (t, CH.sub.2NCO), 6.03 (NHCON), 9.23 (t, NHCO-end group). M.sub.n=2,013 Da, the average polymerization degree (DP)=5.

Example 3: HBP Resin Modified with Amine-Functionalized Tertiary Amine Compound

(12) To a three-necked flask equipped with a reflux condenser, a nitrogen inlet and a connector to a vacuum pump, HBP resin of example 2 (1.5 mmol, comprising 6 mmol blocked isocyanates) was added. After three cycles evacuating and flushing with nitrogen to remove the oxygen, N,N-dimethylethenediamine (DMEN, 27 mmol) was injected to vessel with DMF (30 ml) and stirred at 125° C. for 72 h under nitrogen atmosphere. Then the mixture was concentrated under reduced pressure to half its original volume, dissolved in 30 ml chloroform (CHCl.sub.3) and washed several times with saturated aqueous sodium chloride to remove the excess DMEN, impurities and residual DMF. The organic layer was collected and dried with sodium sulfate. The salt was removed by filtration and all the solvent was removed under reduced pressure. Finally, a slightly reddish resin was obtained (HBP—NH2).

(13) .sup.1H-NMR (400 MHz, DMSO): δ=1.14 to 1.50 (m, CH.sub.2), 2.11 (s, CH.sub.3), 2.21 (t, CH.sub.2N(CH.sub.3)), 2.95, 3.05 and 3.18 (CH.sub.2NCO, CH.sub.2NHCO), 5.60 to 6.02 (NHCON, NHCONH), 9.23 (t, NHCO-end group).

Example 4: HBP Resin Modified with Hydroxide-Functionalized Tertiary Amine Compound

(14) To a three-necked flask equipped with a reflux condenser, a nitrogen inlet and a connector to a vacuum pump, HBP resin of example 2 (1.5 mmol, comprising 6 mmol blocked isocyanates) was added. After three cycles evacuating and flushing with nitrogen to remove the oxygen, 3-(dimethylamino)-1-propanol (DMAP 27 mmol) and tin(II)2-ethylhexanoate (catalyst, a few drops) were injected to vessel with DMF (30 ml) and stirred at 125° C. for 72 h under nitrogen atmosphere. Then the mixture was concentrated under reduced pressure to half its original volume, dissolved in 30 ml chloroform (CHCl.sub.3) and washed several times with saturated aqueous sodium chloride to remove the excess DMEN, impurity and residue DMF. The organic layer was dried with sodium sulfate. After the salt was removed by filtration off all the solvent was removed under reduced pressure. Finally, a slightly reddish resin was obtained (HBP—NH2).

(15) .sup.1H-NMR (400 MHz, DMSO): δ=1.12 to 1.55 (m, CH.sub.2), 1.64 (m, OCH2CH2CH2N), 2.11 (s, CH.sub.3), 2.22 (t, CH.sub.2N(CH.sub.3)), 2.95, 3.07 and 3.18 (CH.sub.2NCO, CH.sub.2NHCO), 3.93 (t, COOCH2), 5.66 to 6.25 (t, NHCON), 7.05 (t, NHCOO).

Example 5: Alkylation of Tertiary Amines

(16) To a solution of tertiary amine-functionalized HBP (0.536 mmol) in dry DMF (5 ml), an alkylating agent (4.8 mmol) was added, and the resulting mixture was stirred overnight with a reflux condenser at different temperature (45° C. for bromoethane, 60° C. for 1-bromobutane and 1-bromohexane, 70° C. for 1-bromooctane, 1-bromodecane, and 1-bromododecane). Next the solution was cool down to room temperature and dropped into diethyl ether. Then the precipitate was dissolved in H.sub.2O and washed with diethyl ether 7 times to remove all the DMF, excess of alkylating agent and other impurities. H.sub.2O phase was collected and dried with freeze-dryer. Finally, yellow waxy solids were obtained (HBP—NH2-Cx or HBP—OH-Cx, in which x is the number of carbon atoms in each alkylating agent). FIGS. 1 and 2 depict the .sup.1H NMR spectra of all products.

Example 6: One-Pot Synthesis of Surfactants

(17) For HBP—NH2-Cx Systems (C6 Exemplified)

(18) To a three-necked flask equipped with a reflux condenser, a nitrogen inlet and a connector to a vacuum pump, bis (hexamethylene) triamine (BHTA, 28 mmol) and carbonyl biscaprolactam (CBC, 56 mmol) were added. After three cycles evacuating and flushing with nitrogen to remove the oxygen, the mixture was dissolved in 20 ml DMF and stirred at 80° C. overnight (≥8 h) under nitrogen atmosphere. Then increasing the reaction temperature to 145° C., the reaction mixture was stirred under N.sub.2 atmosphere for 1 h. After that, the reaction temperature was decreased to 125° C. Meanwhile N,N-dimethylethenediamine (DMEN, 33 mmol) was injected to the flask, and stirred under N2 atmosphere for 72 h. Finally, bromohexane (49 mmol) was injected to the flask, and the reaction was stirred at 70° C. overnight to obtain the final product.

(19) For HBP—OH-Cx Systems (C6 Exemplified)

(20) To a three-necked flask equipped with a reflux condenser, a nitrogen inlet and a connector to a vacuum pump, bis (hexamethylene) triamine (BHTA, 28 mmol) and carbonyl biscaprolactam (CBC, 56 mmol) were added. After three cycles evacuating and flushing with nitrogen to remove the oxygen, the mixture was dissolved in 20 ml DMF and stirred at 80° C. overnight (_8 h) under nitrogen atmosphere. Then increasing the reaction temperature to 145° C., the reaction mixture was stirred under N.sub.2 atmosphere for 1 h. After that, the reaction temperature was decreased to 125° C. Meanwhile 3-(dimethylamino)-1-propanol (DMAP, 33 mmol) and tin(II)2-ethylhexanoate (catalyst, 4 mmol) was injected to the flask, and stirred under N2 atmosphere for 72 h. Finally, bromohexane (49 mmol) was injected to the flask, and the reaction was stirred at 70° C. overnight to obtain the final product.

Example 7: Surfactant Characterization—Critical Micelle Concentration (CMC)

(21) The CMC of the micelles was determined using Nile Red as fluorescence probe. First, 10 mg/ml surfactant/water solution was prepared by weighted 50 mg surfactant to 50 ml Volumetric flask. The suspension was sonicated for 10 min. 50 μL nile red/methanol solution (1 mg/ml) was taken to each Eppendorf tube with the help of Hamilton microsyringe. Methanol was removed at room temperature, then 1 mL surfactant/water solution with different concentration (2.00, 1.50, 1.00, 0.75, 0.50, 0.25, 0.10, 0.075, 0.05, 0.025, 0.01, 0.0005, 0.0001 and 0.00001) was added to each tube. After sharking at 25° C. for 3 h, the mixtures were stored overnight to enable the nile red enter into the micelles. Fluorescence spectra were recorded using a SpectraMax M3 spectrofluorometer with 96-well plates. Emission spectra were recorded from 600 to 750 nm using a λ.sub.exc=560 nm. Excitation and emission slit widths were both maintained at 5.0 nm. In FIG. 3 a representative example is given of the emission spectra of one of the compounds (HBP—OH—C2).

(22) Table 1 shows the CMC of some representative examples.

(23) TABLE-US-00001 TABLE 1 CMC results (mg/ml) for HBP-NH2-Cx and HBP-OH-Cx surfactants HBP-NH2_C.sub.x HBP-OH-Cx Alkyl chain length (mg/ml) (mg/ml) C2 4.7 6.8 C4 3.7 4.9 C6 1.5 3.7 C8 1.2 2.3 C10 0.4 0.5 C12 0.1 0.4

Example 8: Antibacterial Properties

(24) This example demonstrates the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of various antibacterial surfactants of the invention against various bacterial strains.

(25) Preparation of Bacterial Strains

(26) Bacterial strains were cultured from frozen dimethyl sulfoxide stocks on blood agar plates. Subsequently a pre-culture of 10 mL liquid growth medium was inoculated overnight at 37° C. under aerobic conditions. Next, 100 μl of the pre-culture was used to inoculate 10 ml of main culture for 24 h at 37° C. under aerobic conditions. Streptococcus epidermidis ATCC 12228, Streptococcus epidermidis ATCC 12600, Streptococcus epidermidis 1457 or Streptococcus epidermidis ATCC 35984 were all cultured with Tryptone soya broth growth medium and agar (Oxoid, Basingstoke, UK).

(27) In case of bacterial aggregation, main cultures were sonicated 10 sec at 30 W (Vibra Cell model 375, Sonics and Materials Inc., Danbury, Conn., USA) to suspend bacterial dumps. Subsequently, the bacterial concentration was determined using the Bürker Türk counting chamber

(28) Determination of Minimum Inhibitory Concentration (MIC)

(29) A sterile 96 wells plate (Falcon Flat bottom 353072, Tyne & Wear, United Kingdom) was used to mix 200 μl of growth medium containing the bacterial suspension at a final concentration of 105 bacteria/ml with QACs at the final concentrations ranging from 0 to 1280 μg/ml. Concentrations ranges were defined to be both below and above the Critical Micelle Concentration (CMC). Gentamicin at 10 μg/ml was used as a positive control.

(30) The 96 wells plate was incubated for 24 h at 37° C. under aerobic conditions. Bacterial growth was examined visually for each well by assessing changes in turbidity of the suspension after 24 h. The MIC was defined as the well with the lowest QAC concentration for which no growth was observed.

(31) Determination of Minimum Bactericidal Concentration (MBC)

(32) Next, 100 μl of bacterial suspension from the wells that did not show any visual signs of growth was used to inoculate agar plates of the corresponding growth medium. The agar plates were incubated for 24 h and 48 h at 37° C. under aerobic conditions. The MBC was defined as the agar plate inoculated with medium with the lowest QAC concentration for which no growth was observed. Note that the methicillin resistant strains are also resistant for Gentamicin, but not against the compounds of this invention.

(33) TABLE-US-00002 TABLE 2 The antibacterial effect of the surfactant series HBP-NH2-Cx and HBP-OH-Cx, wherein x is 2, 4, 6 8, 10 or 12, against Staphylococcus Epidermidis ATCC 12228. S epidermidis ATCC 12600, S epidermidis 1457, S epidermidis ATCC 35984, S aureus ATCC BAA-1696, A baumanii 1, K pneumoniae 1, E coli ATCC 25922, C albicans GB 1/2, C parapsilosis. S epidermidis ATCC 35984 and S aureus ATCC BAA-1696 are MRSA strains. The concentration of the antibacterial compounds is in μg/mL. S epidermidis S epidermidis S epidermidis S epidermidis S aureus ATCC 12228 ATCC 12600 ATCC 1457 ATCC 35984 ATCC BAA-1696 MIC MBC MIC MBC MIC MBC MIC MBC MIC MBC HBP-NH2-RN.sup.+C.sub.2 2560 >2560 2560 >2560 2560 >2560 2560 >2560 >2560 >2560 HBP-NH2-RN.sup.+C.sub.4 1280 2560 >2560 >2560 2560 2560 640 >2560 2560 >2560 HBP-NH2-RN.sup.+C.sub.6 40 320 160 320 40 320 40 320 80 640 HBP-NH2-RN.sup.+C.sub.8 5 40 10 40 5 40 5 40 20 40 HBP-NH2- 5 20 10 20 5 40 5 10 10 >20 RN.sup.+C.sub.10 HBP-NH2- 5 20 10 20 5 40 5 5 20 20 RN.sup.+C.sub.12 HBP-OH-RN.sup.+C.sub.2 >2560 >2560 >2560 >2560 >2560 >2560 >2560 >2560 >2560 >2560 HBP-OH-RN.sup.+C.sub.4 1280 2560 >2560 >2560 1280 >2560 2560 >2560 2560 >2560 HBP-OH-RN.sup.+C.sub.6 40 320 80 320 40 320 20 160 80 640 HBP-OH-RN.sup.+C.sub.8 5 20 5 40 5 40 5 40 5 40 HBP-OH- 5 5 5 10 5 20 5 10 5 20 RN.sup.+C.sub.10 HBP-OH- 5 5 5 5 5 20 5 5 5 5 RN.sup.+C.sub.12 Gentamidn Inhibition Bactericidal Inhibition Not Inhibition Bactericidal No Not Inhibition Bactericidal 10 μg/ml Bactericidal inhibition bactericidal A baumanii 1 K pneumoniae 1 E coli ATCC 25922 C albicans GB 1/2 C parapsilosis MIC MBC MIC MBC MIC MBC MIC MFC MIC MFC HBP-NH2-RN.sup.+C.sub.2 >2560 >2560 >2560 >2560 >2560 >2560 >2560 >2560 >2560 >2560 HBP-NH2-RN.sup.+C.sub.4 >2560 >2560 >2560 >2560 >2560 >2560 >2560 >2560 >2560 >2560 HBP-NH2-RN.sup.+C.sub.6 >2560 >2560 >2560 >2560 1280 2560 2560 2560 >2560 >2560 HBP-NH2-RN.sup.+C.sub.8 320 320 320 640 80 160 80 >80 640 2560 HBP-NH2- 80 160 80 80 20 40 20 40 160 640 RN.sup.+C.sub.10 HBP-NH2- 40 40 80 80 40 40 20 20 40 160 RN.sup.+C.sub.12 HBP-OH-RN.sup.+C.sub.2 >2560 >2560 >2560 >2560 640 640 >2560 >2560 >2560 >2560 HBP-OH-RN.sup.+C.sub.4 >2560 >2560 >2560 >2560 320 640 >2560 >2560 >2560 >2560 HBP-OH-RN.sup.+C.sub.6 >2560 >2560 >2560 >2560 160 320 1280 1280 >2560 >2560 HBP-OH-RN.sup.+C.sub.8 320 320 160 320 40 80 40 40 320 1280 HBP-OH-RN.sup.+C.sub.10 40 40 20 20 5 20 5 5 20 40 HBP-OH-RN.sup.+C.sub.12 10 10 20 20 5 5 5 5 5 10 Gentamidn No Not No Not Inhibition Bactericidal No Not No Not 10 μg/ml inhibition bactericidal inhibition bactericidal inhibition fungicidal inhibition fungicidal

Example 9: Antimicrobial Coating Compositions

(34) Exemplary high solid water borne polyurethane coating compositions (clear or white) were prepared. In a round bottom flask provided with a dosing funnel and a high speed dissolver (turbo mixer) the ingredients at the relative amounts as shown in Table 3 (clear coating) or Table 4 (white coating) were mixed.

(35) TABLE-US-00003 TABLE 3 Compound function Mass percentage Alberclingk U 9160 80 Byk 024 Defoamer 0.8 HBP-NH2-C8 Antimicrobial surfactant 0.4 Butylcellosolve Solvent 2 Butylcarbitol Coalescent 2 Water 14.2 DSX 1514 Thickener 0.6 100

(36) TABLE-US-00004 TABLE 4 Compound Function Mass percentage Alberdingk U 5201 Resin 62.3 Byk 024 Defoamer 0.8 HBP-NH2-C8 Antimicrobial surfactant 0.6 TiO.sub.2 Kronos 2315 White Pigment paste 25 Dowanol DPM Film forming auxiliary 3 Water Viscosity auxiliary 8 Rheolate 288 Thickener 0.3 Total 100

(37) Stable coating compositions were obtained. A 50 μm thick stable coating was applied on a glass slide with a doctor blade. After drying the coatings at 40° C., smooth coating films were obtained which could withstand 100 acetone double rubs.

Example 10: Antibacterial Coating for Flooring Applications

(38) Different two component reactive coating compositions were prepared using a commercial reference system (blank) with and without addition of an exemplary antimicrobial surfactant of the present invention (HBP—NH2-C8). In a 100 ml container the A component binder was mixed with the dosage of antibacterial agent. After mixing in the centrifuge the appropriate amount of B component isocyanate mixture was added according to ingredients at the relative amounts as shown in Table 5 (PU7550 blank) and Table 6 (PU7550 antibacterial).

(39) TABLE-US-00005 TABLE 5 Compound Function Mass percentage PU 7750 compound A Binder 80 PU 7750 compound B Reactive isocyanate mixture 20 Total 100

(40) TABLE-US-00006 TABLE 6 Compound Function Mass percentage PU 7750 compound A Binder 78.4 HBP-NH2-C8 Antimicrobial surfactant 1.96 PU 7750 compound B Reactive isocyanate mixture 19.6 Total 100

(41) Stable coating compositions were obtained. The resulting mixture was poured onto a seamless floor panel and allowed to harden in a drying cabinet at ambient temperatures. The samples were allowed to fully harden during 21 days, prior to validation of antimicrobial effects.

Example 11: Antibacterial Validation of Reactive Coating Dry Film

(42) An adjusted Japanese industrial standard method (JIS) was applied, samples were placed in a sterile petridish. 100 μL of a bacterial suspensions in PBS (10.sup.6 bacteria/mL) was pipetted on top of a sterile coated sample (cut in pieces 30×30 mm). Next, the well plate was covered with sterilized Parafilm® (24×24 mm) and left to incubate at 37° C. for 24 h under humidified atmosphere. After incubation, 5 mL 0.1% (v/v) Tween80 in PBS was added to each petridish, followed by sonication for 30 s and gentle shaking for 2 min in order to dislodge adhering bacteria. The resulting suspension was serially diluted and the numbers of CFUs were determined by plate counting on agar after 24 h incubation at 37° C., from which the percentage contact-killing efficacy was calculated. Log reduction vs. inoculum values are shown in table 7.

(43) TABLE-US-00007 TABLE 7 Log reduction vs. inoculum Sample S. epidermidis ATCC 12228 Blank seamless floor −3.5 +/− 0.5 PU7550 blank −3.5 +/− 0.5 PU7550 antibacterial  4.0 +/− 0.5

(44) The PU7550 coated sample with the hyperbranched quats as an additive shows a killing effect with a maximum reduction of bacteria on the surface (no bacteria present after inoculation). Both the blank seamless floor and the blank PU7550 coated sample show growth of bacteria, confirming that the observed effect of the sample with hyperbranched quats can be attributed to the antimicrobial additive.

Example 12: Assessing Leaching of Antimicrobial Components

(45) The JIS test was performed, but instead of using bacterial suspension, PBS was used to obtain the same exposure levels of the potential leachable as would be the case in the standard JIS testing. The extract was isolated and stored at 4° C. under N.sub.2. Droplets were pipetted onto agar plates and the agar was fully covered with S. epidermidis and K. pneumoniae. The results are indicated in table 8 showing the presence of an inhibition zone yes/no.

(46) TABLE-US-00008 TABLE 8 Log reduction vs. inoculum S. epidermidis ATCC 12228 K. pneumoniae 1 Sample Inhibition zone Inhibition zone Blank seamless floor No No PU7550 blank No No PU7550 antibacterial No No

(47) The absence of an inhibition zone confirms there are no antimicrobial leachables present in a level able to reduce growth of the strains. This is the case for all of the samples, so no leachable small molecules are related to the observed effects.

Example 13: Composition of Wall Paint with Antimicrobial Additive

(48) Exemplary wall paint coating compositions were prepared. In a paint bucket and using a high speed dissolver (turbo mixer) the ingredients were mixed at the relative amounts as shown in Table 9 (latex formulation), with and without the antibacterial surfactant (hyperbranched quats of the invention, specifically HBP—NH2-C8) or with a reference preservation (isothiazolinones) resulting in 3 samples.

(49) TABLE-US-00009 TABLE 9 Compound Function Mass percentage Orgal PST 50A Latex 14.0 Hecellulose H300 Thickener 0.35 Agitan 282 Defoamer 0.3 HBP-NH2-C8 Antimicrobial 0.5 surfactant TiO.sub.2 TR-92 White Pigment 8.0 Omnyacarb Filler 44.0 Ecodis P90 Dispersing agent 2.0 Water Viscosity auxiliary 29.85 Texanol Coalescing agent 1.0 Total 100

(50) Stable coating compositions were obtained. The coatings were stored and cooled immediately after preparation.

Example 14: Validation of In-Can Preservation of Latex Paints

(51) In can preservation was tested in line with the ISO 11930 protocol. Bacterial strains were collected from the factory environment and identified using Maldi-TOF. The strains were purified and cultured. The latex formulation samples (300 gram each) were contaminated with 1.5 ml of bacterial suspension with a load of 1.5.Math.10.sup.7 cells/gram and split in multiple sterile containers. Controls were prepared without contamination. The test was performed in duplicate. The resulting suspension was serially diluted and the numbers of CFUs were determined by plate counting on agar after 0 days, 1 day, 14 days and 28 days of incubation at 25° C. Results of the enumeration of bacteria (log CFU/g) are shown in table 10.

(52) TABLE-US-00010 TABLE 10 t = 0 t = 1 t = 14 t = 28 days day days days Latex formulation 7.0 7.1 7.4 5.9 No preservation Latex formulation 7.0 7.0 3.5 3.5 Reference preservation Latex formulation 7.0 7.0 2.9 2.9 Antibacterial surfactant of the invention

(53) The results show that the in-can preservation effect of the antibacterial according to the invention is at least similar and even slightly better compared to the reference preservation system using isothiazolinones.

Example 15: Suspension Polymerization

(54) In a flask of 250 mL 5 g MMA, 0.05 g hyperbranched surfactant of the invention (HBP—NH.sub.2—C.sub.8), 1 g benzoyl peroxide and 96 mL water were mixed. The mixture was heated to 80° C., while stirring (2200 rpm) for 6 h (094-A). Under the same conditions 25 g MMA, 0.1 g hyperbranched surfactant (HBP—NH.sub.2—C.sub.8), 1 g benzoyl peroxide and 74 mL water were added and polymerized at 80° C. for 6 h (094-E). The suspensions were stable for at least one month.

Example 16: Comparative Example

(55) In a flask of 250 mL 5 g MMA, 0.05 g polyvinyl alcohol (PVA) 1 g benzoyl peroxide and 96 mL water were mixed. The mixture was heated to 80° C., while stirring (2200 rpm) for 6 h (101-A). Under the same conditions 25 g MMA, 0.1 g polyvinyl alcohol, 1 g benzoyl peroxide and 74 mL water were added and polymerized at 80° C. for 6 h (101-E). The suspension was stable for at least one month.

Example 17: Antibacterial Properties (MIC)

(56) The minimum inhibitor concentrations (MIC) of resulting suspensions of Example 15 and 16 were measured with S. epidermidis (table 11).

(57) The MIC value gives the concentration of biocidal compounds that inhibits growth of bacteria (in a solution of 105 bacteria/mL).

(58) TABLE-US-00011 TABLE 11 MIC values of various polymer suspensions with S. epidermidis ATCC 12228 (10.sup.5 bacteria/mL). Reaction T MIC# Sample MMA (g) Surfactant * ° C. μg/mL 094-A 5 C8 80 8/8 094-E 25 C8 80 8/8 101-A 5 PVA 80 >670 101-E 25 PVA 80 >670 * C8 is hyperbranched surfactant provided with dimethyl, octyl as alkyl groups on the N.sup.+ atom. #MIC = minimum inhibition concentration. The MIC is expressed as μg/mL of the surfactant.

(59) These results demonstrate that the suspension prepared according to the invention, with the surfactant comprising quaternary ammonium moieties results in a inhibition of bacterial growth. In contrast, suspensions prepared under the same conditions with polyvinylalcohol as surfactant do not show any inhibition.

Example 18: Surfactant Provided with Styrene Moiety in Focal Point

(60) The AB.sub.2 monomer (4.79 g, 10 mmol) of example 1 and vinyl benzyl chloride (0.152 g, 1 mmol) were dissolved in xylene. NaHCO.sub.3 (0.5 g) was added as acid scavenger. The solution was heated to 45° C. in a nitrogen atmosphere for 48 h. After salt was removed by filtration, the solution was heat to 145° C. for 2 h.

(61) The solution was cooled down to room temperature and caprolactam was removed by extraction (3×) with an aqueous solution of 1 wt % CaCl.sub.2). The solution was subsequently dried on MgSO.sub.4. N,N,N′-trimethyl-1,3-propanediamine (0.70 g, 6 mmol) was added as functionalize t-amine compound, and the solution was heated for 10 h at 125° C. and cooled down to RT. To this solution 1-bromohexane (0.99 g, 6 mmol) was added and the resulting mixture was stirred overnight with a reflux condenser at 60° C.