WO2003028762A1

WO2003028762A1 - Novel antimicrobial resistance blocking compositions

Info

Publication number: WO2003028762A1
Application number: PCT/US2001/030303
Authority: WO
Inventors: Hanuman B. Jampani; Lester A. Mitscher; Jerry L. Newman; Segagran P. Pillai
Original assignee: Ethicon Inc
Current assignee: Ethicon Inc
Priority date: 2001-09-28
Filing date: 2001-09-28
Publication date: 2003-04-10
Anticipated expiration: 2004-03-28

Abstract

This invention is concerned with antimicrobial compositions that control or prevent resistance to antimirobial effectiveness, more particularly to combinations of topical antimicrobial agents with antimutagenic or antioxidant agents that block intrinsic or acquired bacterial resistance.

Description

NOVEL ANTIMICROBIAL RESISTANCE BLOCKING COMPOSITIONS

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention is concerned with antimicrobial compositions that control or prevent resistance to antimicrobial effectiveness, more particularly to combinations of topical antimicrobial agents with agents that block development of intrinsic or acquired bacterial resistance.

2. Related Art

Among several diseases, opportunistic infectious diseases are leading causes of death among patients. Reputed researchers also suspect a link between certain of these infectious diseases and life threatening diseases like cancer, heart disease, asthma, ulcers and allergies that were not traditionally considered as due to microbial causes. In the United States, an estimated 1.2 million patients each year develop nosocomial infections that are resistant to conventional antibiotics, and approximately 60,000 deaths occur each year adding an estimated $4.5 billion to the annual cost of health care in the US alone. This recent emergence of widespread antimicrobial resistance is a major public health problem locally as well as globally. Antimicrobial resistance by microbes is considered to be intrinsic or acquired. Intrinsic resistance is a natural property of microorganisms. It is often the result of the cell wall or outer membrane restricting entry and intracellular accumulation of antimicrobial agents (including efflux mechanisms) . Also, resistance may be due to biofilms that are inherently resistant to antimicrobials due to sticky conglomerates of bacteria.

Acquired resistance is the result of changes in bacterial genetic material due to mutations or the acquisition of extra chromosomal genetic material. Recent awareness of acquired genetic resistance to triclosan and related antimicrobials through missense mutations of the FABl gene suggests that the widespread use of these types of compounds will lead to increased appearance of resistant microbes that will compromise the present usefulness of triclosan or similar antimicrobial compounds as well as antibiotics used systemically. Researchers clearly suggest that the development of second generation FABl inhibitors would help limit resistance. Certainly, the pharmaceutical industry is working to develop new means of combating resistant pathogens. Diagnostics manufacturers also have an opportunity to develop products that aid treatment by detecting specific drug resistant infections. In addition to focusing on discovering and developing newer antimicrobial compounds, it is very important to better utilize the existing classes of compounds with altogether new strategies to control, prevent or eradicate the resistant organisms. Though frequent handwashing might remove transient flora, dirt and oils from the skin it is becoming increasingly important to formulate unique products with unique technological characteristics that are capable of enhancing antimicrobial effectiveness by removing/killing antibiotic resistant strains while preventing the emergence of resistant strains from susceptible strains.

Resistance to antimicrobial agents is now a serious clinical and a major public health problem in the U.S. The international spread of pathogenic organisms clearly demands a global strategy. New drugs are unlikely to appear soon enough and in sufficient number to solve many of these resistance problems. Hence, there is a growing need to understand the factors that lead to evolution of the spread of resistance and to design strategies to maximize the effectiveness of existing and future drugs/products while minimizing the spread of resistance to them. A recent article in the American Journal of Infection Control (AJIC, 26:541-3 (1998)) illustrates the continuing need to examine various facets of the problem and suggested solutions . The antimicrobial compositions of this invention offer one solution to this problem. SUMMARY OF THE INVENTION

The present invention relates to a topical antimicrobial composition comprising:

(a) a topical antimicrobial agent; and

(b) an antimicrobial resistance blocking effective amount of at least one antimutagenic and/or antioxidant agent .

In one embodiment, the topical antimicrobial is triclosan and the antimutagenic and/or antioxidant compound is selected from the group consisting of substituted and unsubstituted pyrithione-containing compounds; coumarins; pseudopeptides; indazoles; antioxida ts; flavanoids; isoflavanoids including isoflavenes, isoflavanes, isoflavanones and isoglabrene analogs; and mixtures thereof.

Another aspect or embodiment of the invention provides a method for inhibiting bacterial resistance comprising:

(a) incorporating an antimicrobial resistance blocking effective amount of at least one antimutagenic and/or antioxidant agent into a topical antimicrobial agent; and

(b) applying the composition of (a) to a surface containing bacteria. DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS OF THE

INVENTION

As used herein, the term "topical antimicrobial" is intended to describe compounds used to inhibit or kill or otherwise prevent proliferation of microbes . The topical antimicrobial agents are used to treat any type of surface, both mammalian and non-mammalian. In humans, these agents are widely used for treatment of the skin.

Suitable topical antimicrobial agents for use with this invention include nisin, bis-diguanides, chlorhexidine gluconate, chlorhexidine digluconate, chlorhexidine diacetate, chlorhexidine dihydrochloride, polyhexamethylene biguanide, benzalkonium chloride, benzethonium chloride, methylbenzethonium chloride, cetyl pyridinium chloride, triclosan, triclocarban, tribromosilane, amyltricresols, parachlorometaxylenol, phenol, silver, iodine, [nonylphenoxypoly (ethyleneoxy) ethanoliodine] poloxamer-iodine complex, undecoylium chloride, iodine complex, bisquaternary ammonium compounds, polymeric quaternary ammonium compounds, alcohols, cationic polypeptides, organometallic antiseptics, alkyl pyridinium salts, essential oils, and their combinations and derivatives. Likewise, a variety of simple alcohols may function in this regard, including but not limited to, ethanol, propanol, butanol, pentanol, 2-methyl-l-butanol, hexanol, 2- methyl-1-pentanol, 3-methyl-1- pentanol, 2-ethyl-l- butanol, 3 , 5 , 5-trimethyl-l-hexanol, heptanol, octanol, isooctyl alcohol, decanol, dodecanol, tridecanol, tetradecanol and the like.

The term "antimutagenic" compound or agent, as used herein, is intended to describe compounds that lessen the extent of antimicrobial resistance. As will be shown hereinafter, not all antimutagenic agents are effective in inhibiting antimicrobial resistance when combined with topical antimicrobial agents.

As used herein, the term "antioxidant" compound or agent is intended to describe oxygen scavenging compounds that prevent or lessen the degree of mutations in genetic material (e.g., DNA) , or prevent or lessen the degree of oxidation of poly-unsaturated fatty acids.

Also as described hereinafter, certain suitable antimutagenic and antioxidant agents are described both by chemical name and chemical structure. Due to the fact that chemical nomenclature for the same structure can vary and for the avoidance of doubt any discrepancy that may arise between a chemically named compound and the corresponding chemical structure shown, the chemical structure is intended to govern the description of the compound. Examples of suitable antimutagenic and antioxidant compounds useful in this invention include the following compounds and analogs thereof: substituted and unsubstituted pyrithione-containing compounds; coumarins; pseudopeptides; indazoles; antioxidants; flavanoids; isoflavanoids including isoflavenes, isoflavanes, isoflavanones and isoglabrene analogs; and mixtures thereof.

Examples of pyrithione-containing compounds include substituted and unsubstituted pyrithione containing compounds such as monovalent or divalent metallic salts of pyrithione. Preferred are zinc, silver, and sodium salts of pyrithione.

Zinc Pyrithione Sodium Pyrithione Examples of suitable coumarins include: 8- (2'2'-Dimet yl-l'H-pyran-7-yl) coumarin; [ (2 '2-Dimet yl-3-β- ydroxy-6-methylene) -1-β-cyclohexyl] -3 -methyl - pent-2-enyl-7-oxycoumarin; [ (2'2,6-Trimethyl-2-oxo-bicyclo)-2,2,l-heptyl]-l-β-3-methyl-pent-2- enyl-7-oxycoumarin; 7-Cyclohexylmethoxycoumarin; and Galbanic Acid.

[(2'2-Dimethyl-3-β-hydroxy-6-methylene)-l-β-cyclohexyl]- -(2'2'-Dimethyl- 1 H-pyran-7-yl)coumarin 3-methyl-pent-2-enyl-7-oxycoumarin

[(2'2,6-Trimethyl-2-oxo-bicyclo)-2,2,l-heptyl]l-β-3-methyl-pent-2-enyl-7-oxycouιnaιin

7-Cyclohexylmethoxycoumarin

Galbanic Acid

Preferred coumarins are :

8- (2 ' 2 ' -Dimethyl-l Η-pyran-7-yl ) coumarin; and Galbanic Acid.

Most preferred coumarins are :

[ ( 2 ' 2-Dimethyl-3-β-hydroxy-6-methylene) -1-β-cyclohexyl] -3-methyl - pent-2-enyl-7-oxycoumarin; [ ( 2 ' 2 , 6-Trimethyl-2-oxo-bicyclo) -2 , 2 , 1-heptyl] -l-β-3-methyl-pent-2- enyl-7-oxycoumarin; and 7-Cyclohexylmethoxycoumarin .

Examples of suitable pseudopeptides include :

N-Tert-butoxycarbonyloxyaminopentan-N, N-diethylamide; N-Benzoylalanyl-N, N-diethylamide ; N-Tert-Butoxycarbonyloxyalanyl~N, N-diethylamide ; N-Tert-Butoxycarbonyloxyalanyl-N, N-morphilinoamide; N-Tert-butoxycarbonyloxyalanyl-N-methyl-N-phenylamide; 4 -Benzoylaminobenzoyl-N, N-diethylamide; 4-Benzyloxycarbonyloxybenzoyl-N, N-diethylamide; 4-Toluenesulfonylaminobenzoyl-N, N-diethylamide , • 4-Benzoylaminobenzoyl-N, N-diethylamide; 4-Benzyloxycarbonyloxybenzoyl-N, N-diethylamide ; 5-N-Benzylaminopentanoyl -N, N-diethylamide , • 5 -N-Cyclopentylaminopentanoyl-N, -diethylamide; and 5-N-Benzoylaminopentanoyl-N, N-diethylamide . Preferred pseudopeptides are :

N-Tert-butoxycarbonyloxyaminopentan-N, N-diethylamide; N-Benzoylalanyl-N, N-diethylamide ; N-Tert-Butoxycarbonyloxyalanyl-N, N-diethylamide; N-Tert-Butoxycarbonyloxyalanyl-N, N-morphilinoamide; N-Tert-butoxycarbonyloxyalanyl-N-methyl-N-phenylamide ; 4 -Benzoylaminobenzoyl-N, -diethylamide ; 4 -Toluenesulfonylaminobenzoyl-N, N-diethylamide; and 4-Benzoylaminobenzoyl-N, N-diethylamide .

Most preferred pseudopeptides are :

4-Benzyloxycarbonyloxybenzoyl-N, N-diethylamide;

5-N-Benzylaminopentanoyl-N, N-diethlamide;

5 -N-Cyclopentylaminopentanoyl-N, N-diethylamide; and

5-N-Benzoylaminopentanoyl-N, N-diethylamide .

N-tert-butoxycarbonyloxyamιnopentan-N,N-dιethylamιde

N-Benzoylalanyl N,N-dιethylamιde

N-tert-butoxycarbonyloxyalanyl-N-methyl-N-phenylamide 4-Benzoylamιnobenzoyl-N,N-dιethylamιdι

4-Benzyloxycarbonyloxybenzoyl-N,N-dιethylamιde 4-Toluenesulfonylamιnobenzoyl-N,N-dιethyla)

4-Benzyloxycarbonyloxybenzoyl-N,N-dιethylamιde

4-Benzoylamιnobenzoyl-N,N-dιethylamιde

5-N-Benzylamιnopentanoyl-N,N-dιethylamιde 5-N-Cyclopentylamιnopentanoyl-N,N-dιethylamιde

5-N-Benzoylamιnopentanoyl-N,N-dιethylamιde

BOC = tert/butoxycarbonyl Me = CH₃ Examples of suitable indazoles include :

1 -Methyl -3 -benzyl oxy-5-ni tro-lH-indazole ; l-Methyl-3 - [ ( 3-dimethylamino) propyloxy] -lH-indazole;

1 -Benzyl -3 -ethoxy- 5 -ni tro-lH-indazole; l-Methyl-3-ethoxy-5-benzoylamino-lH-indazole; l-Methyl-3 -ethoxy-5-benzylaminocarboxylamino-lH-indazole; l-Methyl-3 -tosyloxy-5-ni tro-lH-indazole ; l-Methyl-3 -bromo-5-nitro-lH-indazole; l-Methyl-3-benzyloxy-5-amino-lH-indazole; l-Phenyl-3-hydroxy-5-nitro-lH-indazole; l-Benzyl-3-benzyloxy-5-nitro-liϊ-indazole; l-Methyl-3-benzyloxy-5-nitro-liϊ-indazole; l-Methyl-3- [ (3-dimethylamino) propyloxy] -5-amino-l£f-indazole; l-Benzyl-3-ethoxy-5-amino-lH-indazole;

Benzydamine l-Methyl-3- [ (dimethylamino) propyloxy-lH-indazole;

Isoniazid or 4-pyridinecarboxylic acid hydrazide; l-Methyl-3-ethoxy-5-nitro-lH-Indazole; and l-Methyl-3 -ethoxy-5-amino-liT-Indazole .

Preferred indazoles are :

l-Methyl-3 -benzyl oxy-5 -nitro-lff-indazole ; l-Benzyl-3-ethoxy-5-nitro-lH-indazole; l-Methyl-3-ethoxy-5-benzoylamino-lH-indazole; l-Methyl-3-ethoxy-5-benzylaminocarboxylamino-lH-indazole;

1-Methyl-3-tosyloxy-5-nitro-lH-indazole; l-Methyl-3-benzyloxy-5-amino-lH-indazole; l-Methyl-3- [ (3-dimethylamino) propyloxy] -5-amino-lff-indazole; l-Benzyl-3-ethoxy-5-amino-lH-indazole; and

Isoniazid or 4-pyridinecarboxylic acid hydrazide; Most preferred indazoles are :

l-Methyl-3- [ ( 3-dimethylamino) propyloxy] -lH-indazole; l-Methyl-3-bromo-5-nitro-lH-indazole; l-Phenyl-3-hydroxy-5-nitro-lH-indazole; l-Benzyl-3-benzyloxy-5-nitro-lH-indazole; l-Methyl-3 -benzyloxy-5-nitro-lH-indazole; l-Methyl-3 - [ (dimethylamino) propyloxy-lH-indazole; l-Methyl-3-ethoxy-5-nitro-lH-Indazole; and l-Methyl-3-ethoxy-5-amino-liϊ-Indazole .

l-Methyl-3-benzyloxy-5-nitro-lJ7-indazole l-Methyl-3-[(3-dimethylamino)propyloxy]-l#-indazole

1 -Methyl-3-ethoxy-5-benzoylamino- 1/7-indazole l-Benzyl-3-ethoxy-5-nitro-li7-indazole

l-Methyl-3-ethoxy-5-benzylarninocarboxylamino-lfl'-indazole l-Methyl-3-tosyloxy-5-nitro-li?-indazole

l-Methyl-3-bromo-5-nitro-l t-indazole l-Methyl-3-benzyloxy-5-arruno #-indazole

2-Hydroxy-4-mesyloxybenzaldehyde 2'-0-Methyl-7-hydroxyisoglabrene

Et = CH₂CH₃ Me = CH₃ Ph = Phenyl l

l-Benzyl-3-benzyloxy-5-nitro-lH-indazole

l-Methyl-3-benzyloxy-5-nitro-l#-indazole l-Methyl-3-[(dimethylamino)propyloxy-lfl^r-inda₂

l-Methyl-3-ethoxy-5-nitro-liϊ-indazole l-Methyl-S-benzyloxy-S-nitro-liϊ-indazole

l-Methyl-3-[(3-dimethylamino)propyloxy]-5-amino-liϊ-indazole l-Benzyl-3-ethoxy-5-amno-l/7-indazo]

Me = CH₃ p_{h =} Phenyl _Et=CH₂CH₃ Examples of suitable isoglabrene analogs include :

4 ' -Methoxyisof lav-3-ene;

4 ' -Hydroxyisof lav-3-ene;

2 ' -Methoxyisof lav-3-ene;

7 , 2 ' -Dimethoxyisoflav-3-ene;

2 , 4 ' -Dimethoxyisoflav-3-ene;

7-Methoxy-3- [5-methoxy-2 , 2-dimethyl-3 , 4-dihydro-2H-l-benzopyran6- yl] 2-H- ] -benzo-3-pyran;

2 ' O-Methylisoglabrene; Isoglabrene;

2 ' -O-Methyl-3 , 4, 3 "4"-tetrahydroisoglabrene;

3 "4 " -Dihydroisoglabrene;

2 ' 0-Methyl-7-demethyl-7-methanesulfonylisoglabrene;

7-Hydroxy-6-aceto-2,2-dimethylbenzo-lH-benzopyran;

5-Hydroxy-6-acetyl-2 , 2-dimethyl-2H-l-benzopyran;

5-Methoxy-6- [ (2-hyroxymethyl-5-mesyloxy) -2-phenyloxy]aceto-2, 2- dimethylbenzo-lH-benzopyran; and

2-hydroxy-4-mesyloxybenzylacohol .

Preferred isoglabrene analogs are:

4 ' -Hydroxyisoflav-3-ene;

7,2' -Dimethoxyisoflav-3-ene;

7-Methoxy-3- [5-methoxy-2 , 2-dimethyl-3 , 4-dihydro-2H-l-benzopyran6- yl] 2-H-] -benzo-3-pyran;

2 ' -O-Methyl-3 , 4 , 3 "4 " -tetrahydroisoglabrene;

5-Hydroxy-6-aceryl-2, 2dimethyl-2H-l-benzopyran;

2-hydroxy-4-mesyloxybenzylacohol; and

2 ' 0-Methyl-7-demethyl-7-methanesulfonylisoglabrene. Most preferred isoglabrene analogs are :

2 ' -Methoxyisof lav-3-ene ;

2 , 4 ' -Dimethoxyisoflav-3-ene ;

2 ' O-Methylisoglabrene;

Isoglabrene;

3 " 4 " -Dihydroisoglabrene ;

2 ' 0-Methyl-7-demethyl-7-methanesulf onylisoglabrene;

7-Hydroxy-6-aceto-2 , 2-dimethylbenzo-lJϊ-benzopyran; and

5-Methoxy-6- [ (2-hyroxymethyl-5-mesyloxy) -2-phenyloxy] aceto-2 , 2- dimethylbenzo-lH-benzopyran .

- 18 -

3 "4"-Dihydroisoglabrene 4'-Hydroxyisoflav-3-ene

7,2'-Dimethoxyisoflav-3-ene '-Methoxyisoflav-3-ene

zo]

,4'-Dimethoxyisoflav-3-ene

2'-<9-Methylisoglabrene

Isoglabrene

2'-0-Methyl-3 ,4,3 "4"-tetrahydroisoglabrene 3 "4"-Dihydroisoglabrene

2-O-Methyl-7-demethyl-7-methanesulfonylisoglabrene 7-Hydroxy-6-aceto-2,2-dimethylbenzo-liϊ-

5-Hydroxy-6-acertl-2,2-dimethyl-2iϊ-l-benzopyran 5-Methoxy-6-[(2-hyroxymethyl-5-mesyloxy)-2- aceto-2,2-dimethylbenzo-l t-benzopyran

Suitable antioxidants include trolox, reduced glutathione, butylated hydroxyanisole (BHA) , butylated hydroxytoluene (BHT) , (-) -Epigallocatechin Gallate

(EGCG) , (-) -Gallocatechin Gallate (GCG),

(-) -Epigallocatechin (EGC) , (-) -Epicatechin Gallate

(ECG) and (-) -Epicatechin (EC).

Preferred antioxidants are trolox, BHA, BHT, Gallocatechin, Gallate, Epigallocatechin, Epicatechin Gallate and Epicatechin.

Most preferred antioxidants are reduced glutathione and Epigallocatechin Gallate.

(-)-Epicatechin Gallate [(-)-ECG] (-)-Epigallocatechin Gallate [(-)-ECCG]

(-)-Epigallocatechin [ECG]

(-)-Gallocatechin Gallate [GCG]

(-)-Epicatechin [(-)-EC]] The combination of topical antimicrobials and the antimutagenic and/or antioxidant compounds requires that the antimutagenic and/or antioxidant compounds be used in an effective amount. As used herein, the term effective amount refers to that amount of antimutagenic and/or antioxidant compound that when combined with the topical antimicrobial agent decreases the amount of antimicrobial resistant colonies present in the antimicrobial by at least 20%. Thus, if a topical antimicrobial measures a antimicrobial resistant colony (RC) value of, for example, RC=20, the effective amount of antimutagenic and/or antioxidant compound added would need to reduce antimicrobial resistant colony value equal to RC=16 or lower. As will be shown later, not all antimutagenic and/or antioxidant compounds are effective in reducing antimicrobial resistant colonies in topical antimicrobials.

Why any effective amount of the antimutagenic and/or antiodoxidant compounds may be used and may widely vary, typical effective amounts when the compound is a pyrithione-containing compound will range from 0.1 to 25, preferably from 0.1 to 10, most preferably from 0.1 to 5 μg/ml on an individual basis. Other antimutagenic and/or antioxidant compounds may be used in effective amounts ranging from 1 to 50, preferably from 1 to 25, most preferably from 1 to 10 mg/ml on an individual basis. The relative ratio of topical antimicrobial to antimutagenic and/or antioxidant useful in this invention ranges generally from 1: 0.1-50, preferably from 1 : 0.1-10.0 , more preferably from 1 : 0.5-5.0, and most preferrably from 1:1, and vice versa based on weight.

The antimicrobial resistance blocking compositions of this invention may be used in a wide variety of applications . Such applications include antimicrobial skin care products, antimicrobial wound dressings, antimicrobial therapeutic gels, anticancer compositions, antimicrobial gloves, antimicrobial skin preparations, antimicrobial drapes, antimicrobial scrubs, antimicrobial gels, antimicrobial lotions, antimicrobial contact lenses, antimicrobial artificial skin grafts, antimicrobial gene delivery systems, antimicrobial polypeptide and antimicrobial household products to name a few.

The antimicrobial resistance blocking compositions of this invention may be included with other skin treatment additives. Examples of suitable additives include: skin protectants, anti-fungal compounds, surface active materials, cationic antimicrobials, natural oils, plant and marine derived bioactive and/or natural products, phospholipids, liposomes, cyclodextrins and metal oxides (e.g., zinc, copper).

EXAMPLES

The present invention may be further understood by the following examples which are intended only to be illustrative and not restrictive of the present invention.

Materials and Methods

Laboratory based bacterial and yeast strains

The following bacterial strains were obtained from the American Type Culture Collection and were used in establishing antimicrobial and antibiotic sensitivity levels. Staphylococcus aureus ATCC 13709, Escherichia coli ATCC 9637, Salmonella choleraesuis ATCC 9184, Pseudomonas aeruginosa ATCC 27853, Klebsiella pne moniae ATCC 10031, and Candida albicans ATCC 10231 Escherichia coli AG100 was obtained through the courtesy of Professor Stuart Levy of Tufts University, Boston, MA.

Chemicals

Triclosan (Irgasan DP300) was obtained from CIBA Specialty Chemical Corp. To evaluate the degree of development of antimicrobial resistance, growth assays were performed using a selection of laboratory based bacteria in a 96 well ELISA plate format with varying concentrations of antiseptics and antibiotics in Oxoid Nutrient Broth No. 2 at 37°C. Readings were taken at 570 nm every 24 hours for 12 days using a Cambridge Technology, Inc., Plate Solver Ver. 4.00 and graphed as a function of growth vs. time. A second set of experiments used resistant strains of bacteria subcultured from survivors of the first experiment.

MIC and MBC assays

MIC and MBC assays were performed using various laboratory strains and multiple antibiotic resistant strains of bacteria collected from patients. The strains were grown in Oxoid Nutrient Broth No. 2 using a 96 well ELISA plate containing varying concentrations of test compounds with incubation for 15 hours at 37°C. After incubation the plates were examined using a Cambridge Technology, Inc., Plate Solver Ver. 4.00 at 570 nm. MIC-100 values were assigned from wells showing no absorbence. Following this, the media from wells showing no growth were streaked on Oxoid Nutrient Agar No. 2 plates and incubated for 15 hours at 37°C. MBC-100 values were assigned from plates showing no visible growth. Antibiograms (Kirby-Bauer) agar diffusion method for determination of zones of inhibition.

Sterile blank disks (6 mm in diameter) were impregnated with 5 μl on 10 μg/ml solutions of a variety of antimicrobials. Incubation was at 37°C for four days on Oxoid Nutrient No. 2 Agar plates. The plates were read on the second day for determinations of the diameter of the inhibition zones and on the fifth day for resistance development data. Resistance was measured by estimating the percentage of the zone of inhibition that was covered by resistant colonies or, in other experiments, by counting the number of individual colonies that developed in the otherwise clear zone of inhibition.

Example 1 - Bacterial Resistance to Triclosan

When standard strains of bacteria were incubated for fourteen days in enriched media containing graded concentrations of triclosan ranging in two-fold increments from 0.25-256 μg/ml, Escherichia coli AG100 strain was controlled for 13 days by doses of 2 μg/ml or more, but breakthrough to resistance was seen at concentrations between the MIC and MBC values in the form of perceptible growth in 9 days with 1 μg/ml and at 5 days with 0.5 and 0.25 μg/ml of triclosan. Escherichia coli ATCC 9637 was controlled by doses of triclosan of 0.5 μg/ml and above but developed resistance in 5 days at 0.25 μg/ml. With Klebsiella pneumoniae ATCC 10031, resistance breakthrough was seen at 9 days with 2 μg/ml, at 5 days with 1 μg/ml, at 4 days with 0.5 μg/ml and 0.25 μg/ml. Candida albicans ATCC 10231 developed resistance in 2 days at 2 μg/ml. S. aureus ATCC 13709 was controlled for 14 days by all concentrations employed, but Pseudomonas aeruginosa ATCC 27853 and Salmonella choleraesuis ATCC 98184 were unaffected by any of these doses (Table 1-1) .

TABLE 1-1

Culture Breakthrough Breakthrough time level s. aureus ATCC 13709 — None

E. coli AG 100 9 days lμg/ml 5 days 0.5μg/ml* 5 days 0.25μg/ml**

E. coli ATCC 9637 5 days 0.25μg/ml**

K. pneumoniae ATCC 10031 9 days 2μg/ml* 5 days lμg/ml* 4 days 0.5μg/ml* 4 days 0.25μg/ml

C. albicans ATCC 10231 2 days 2μg/ml

P. aeruginosa ATCC 27853 Not sensitive up to 256μg/ml S. choleraesuis ATCC 9184 Not sensitive up to 256μg/ml

Table 1-1. Breakthrough levels to resistance by a series of laboratory microorganisms exposed to incremental doses of triclosan for 14 days, * indicates the growth did not reach the same level as control growth reached. ** indicates that growth reached the same level as the control growth reached but at the 11^th day of incubation. Generally the controls reached maximal growth levels by day 2.

Colonies of E. coli ATCC 9637 and K. pneυmoniae

ATCC 10031 shown to be resistant to triclosan in this manner failed to be inhibited by triclosan at any concentration between 0.25 and 256 μg/ml. On the other hand, triclosan resistant C. albicans ATCC 10231 was less resistant, showing growth in 5 days at 1 μg/ml, in 3 days at 0.5 μg/ml, and in 2 days at 0.25 μg/ml (Table 1-2) . The C. albicans results suggest persistence rather than resistance.

TABLE 1-2

Culture Breakthrough Breakthrough time level

E. coli ATCC 9637 R at Insensitive from 0 . 25 -

0.25μg/ml 256μg/ml K. pneumoniae ATCC 10031 Insensitive from 0 . 25 -

R2μg/ml 256μg/ml C. albicans ATCC 10231 5 days lμg/ml

Rlμg/ml 3 days 0 . 5μg/ml 2 days 0 . 25μg/ml

Table 1-2 . Breakthrough levels to resistance by triclosan resistant bacteria when subsequently exposed to incremental doses of triclosan for 14 days . (R . figures indicate the concentration of triclosan from which these resistant colonies were cultured . )

The MIC-100 and MBC-100 values for triclosan against these ATCC strains and E. coli AG 100 were determined by agar dilution methods and these are listed in Table 1-3 . The difference between these values is 2- 8 fold indicating that bacteriocidal doses of triclosan can easily be reached through the use of comparatively modest concentrations, except with Pseudomonas aeruginosa. and Salmonella choleraesuis which are intrinsically highly resistant at the outset. Repetition of these measurements using strains deliberately made triclosan resistant showed that E. coli and K. pneumoniae became highly resistant, but the C. albicans strain was still sensitive to triclosan. These results are in general agreement with those in Tables 1-1 and 1-2.

TABLE 1-3

Culture Triclosan Triclosan-resistant

MIC MBC MIC MBC

S. aureus ATCC 13709 0.25 1 — —

E. coli ATCC 9637 0.25 0.5 >256 >256

E. coli AG 100 0.5 4 — --

K. pneυmoniae ATCC 10031 0.25 4 >256 >256

C. albicans ATCC 10231 2 4 2 4

P. aeuruginosa ATCC 27853 >256 >256 -- --

S. choleraesuis ATCC 9184 >256 >256

Table 1-3 . A comparison of the MIC-100 and MBC-100 values (in μg/ml ) for triclosan naϊve and triclosan resistant strains of various bacteria .

Example 2 - Zinc Analogs

This example shows that when zinc pyrithione or sodium pyrithione are added to triclosan, the combination produces broad spectrum suppression of triclosan resistance , with the exception of Klebsiella pneυmoniae . A variety of zinc containing analogs were examined and only zinc pyrithione gave useful MIC and MBC values when they were examined individually (Table 2-1) . As noted in Table 2-1 's footnote, zinc chloride, zinc sulfate, zinc acetate and zinc metal powder failed to inhibit. Sodium pyrithione gave somewhat less impressive MIC-100 values than zinc pyrithione and was ineffective at large doses in the MBC studies except, for S, aureus and C. albicans.

TABLE 2-1

Culture Zinc pyrith.ione Sodium pyrithione

MIC- 100 MBC- -100 MIC-100 MBC-100

S. aureus ATCC 13709 0.5μg/ml 2μg/ml lμg/ml 2μg/ml

E. coli AG100 0.5 4 4 >256

E. coli ATCC 9637 8 8 8 >256

K. pneumoniae ATCC 10031 2 4 8 >256

C. albicans ATCC 10231 0.5 2 0.25 4

P. aeruginosa ATCC 27853 32 32 64 >256

S. choleraesuis ATCC 9184 32 32 64 >256

Table 2-1. MIC-100 and MBC-100 levels for zinc pyrithione and sodium pyrithione against a variety of microorganisms . When tested under the same conditions , zinc chloride, zinc sulfate, zinc acetate and zinc metal powder were inactive below 128 μg/ml .

When zinc pyrithione and sodium pyrithione were added individually to disks also containing an equal concentration of triclosan, resistance development (as judged from the presence of no colonies within the zone of inhibition) was completely prevented in E. coli ATCC9637 and AG 100 but these compounds failed to provide the same protective effect against K. pneumoniae ATCC 10031. No other zinc salt tested showed this effect (Table 2-2). Given its more bacteriocidial effect and somewhat broader spectrum, the combination of zinc pyrithione and triclosan is to be preferred.

TABLE 2-2

Culture Triclosan Zinc pyrithione Triclosan +

Zinc pyrithione

S. aureus ATCC 13709 54 mm (0) 15 mm (0) 57 mm (0)

E. coli AG100 31 (15) 18 (0) 35 (0)

E. coli ATCC 9637 28 (15) 14 (0) 28 (0)

K. pneumoniae ATCC 10031 35 (18) 14 (0) 35 (11)

C. albicans ATCC 10231 21 (0) 26 (0) 30 (0)

P. aeruginosa ATCC 27853 0 0) 7 0) 9 (0)

S. choleraesuis ATCC 9184 0 0) 10 (0) 12 (0)

Culture Triclosan Sodium pyrithione Triclosan H

Sodium pyrithione

S. aureus ATCC 13709 49 mm (0) 29 mm 48 mm (0)

E. coli AG100 31 (13) 33 (0) 31 (0)

E. coli ATCC 9637 28 (13) 26 (0) 31 (0)

K. pneumoniae ATCC 10031 35 (15) 29 0) 35 (15)

C. albicans ATCC 10231 24 (0) 42 (0) 42 (0)

P. aeruginosa ATCC 27853 0 : o ) 0 :o) 0 (0)

S. choleraesuis ATCC 9184 0 0) 0 : o j 0 (0)

Table 2-2. Effect of zinc and sodium pyrithione at 10 μg/ml on the zone sizes obtained for triclosan at the same concentration and on the development of resistant colonies in the resulting zones of inhibition. Cultures were incubated for 5 days at 37°c. The number of resistant colonies that developed are listed in the parentheses. Zinc acetate, zinc chloride, zinc sulfate and zinc metal powder were ineffective in producing zones or in reducing the number of resistant colonies that developed. Interestingly, however, breakthrough to resistance could be observed when all of the cultures, except P. aeruginosa and S. choleraesuis, were incubated for a long time (14 days) with zinc pyrithione (Table 2-3).

TABLE 2-3

Culture Breakthrough Breakthrough time level

S. aureus ATCC 13709 4 days lμg/ml*

4 days 0.5μg/ml*

E. coli AG 100 7 days 2μg/ml*

3 days lμg/ml*

2 days 0.5μg/ml*

2 days 0.25μg/ml*

E. coli ATCC 9637 11 days 4μg/ml*

8 days 2μg/ml*

5 days lμg/ml*

3 days 0.5μg/ml*

2 days 0.25μg/ml*

K. pneumoniae ATCC 10031 4 days 2μg/ml*

C. albicans ATCC 10231 4 days lμg/ml*

2 days 0.5μg/ml*

P. aeruginosa ATCC 27853 — Not sensitive up to 32μg/ml

S. choleraesuis ATCC 9184 — Not sensitive up to 32μg/ml

Table 2-3. Breakthrough levels to resistance by a series of laboratory microorganisms exposed to incremental doses of zinc pyrithione for 14 days . * indicates that growth did not reach the same level as control growth reached. Generally the controls reached maximal growth levels by day 2.

Thus, in view of Examples 1 and 2, it is not difficult to demonstrate in the laboratory that certain common bacteria produce resistant or persistent colonies when exposed to insufficient concentrations of triclosan. In our hands, in addition to E. coli AG100 for which resistance has been described by others, ATCC strains of E. coli, K. pneumoniae, and C. albicans also produce such colonies as summarized in Tables 1-1 and 1- 2. E. coli AG100 is significantly more prone to produce such colonies than E. coli ATCC strain 9637« K. pneumoniae ATC 10031 likewise produces resistant colonies over a ten-fold concentration range. These concentrations are significantly smaller than those used in triclosan preparations unless they are highly diluted. When picked and regrown in rich media, the triclosan-resistant colonies of E. coli and K. pneumoniae, however, were found to be very resistant to triclosan (Table 1-2). The concentrations required to lyse the resistant colonies may well lie outside the concentrations one is advised to use. Interestingly, resistant colonies of C. albicans remained sensitive to triclosan under the same conditions. P. aeruginosa and S. choleraesuis were resistant from the outset, but S. aureus was sensitive to triclosan and did not develop resistant colonies in these experiments. It seems therefore, advisable to use triclosan preparations at significant concentrations so as to avoid the emergence of resistant strains.

In other experiments the MIC-100 and MBC-100 values were determined. Triclosan-resistant E. coli and K. pneumoniae were 1000-fold less sensitive to triclosan whereas triclosan-resistant C. albicans retained its sensitivity values (Table 1-3) .

These methods should be useful in detecting other agents that have similar useful activity. The data in Table 2-1 show that zinc and sodium pyrithione, an antiinfective agent frequently used in cosmetics, are active against both the Gram positive and Gram negative bacteria used in this study. The use of the pyrithione salts along with triclosan resulted in the elimination of resistant colony development, once again with the exception of K. pneumoniae (Table 2-2) . Since zinc pyrithione is bacteriocidal at much smaller doses than the sodium salt, it should provide a smaller opportunity for resistant colony development and is the preferred salt to use.

Example 3 - Coumarins

This example summarizes the effectiveness of coumarins on the prevention of Triclosan (IRGASAN DP300) antimicrobial resistance development in E. coli ATCC 9637.

Table 3A shows the effect of Galbanic acid analogs on the prevention of Triclosan (IRGASAN DP300) antimicrobial resistance development in E. coli ATCC 9637. In this case, 5μl of lOmg/ml solution was embedded for each compound on a 6mm sterile disk.

The diameter of the zone of inhibition was measured in mm after incubation for 5 days at 37°C. The number of resistant colonies within the zone of inhibition was recorded down as RC values .

TABLE 3A

Strain E. coli ATCC 9637

Table 3B shows the effect of coumarin analogs on the prevention of Triclosan (IRGASAN DP300) antimicrobial resistance development in E. coli ATCC 9637. In this case, 5μl of lOmg/ml solution was embedded for each compound on a 6mm sterile disk. The diameter of the zone of inhibition was measured in mm after incubation for 5 days at 37°C. The number of resistant colonies within the zone of inhibition was recorded down as RC values .

4'4Dihydroxy-3,3'Dimethoxy-Dicinnamyl ester [(2'2-Dimethyl-3-β-hydroxy-6-methyl-5-deh β-cyclohexyl]-3-methyl-pent-2-enyl-7-oxycc

[(2'2-Dimethyl-3-β-hydroxy-6-β-hydroxy-6-methyl-l-β-cyclohexyl]-3-methyl-pent-2-enyl-7-oxycourn

TABLE 3B

Strain E. coli ATCC 9637

As can be seen from Tables 3 A and 3B, the results show that only the combinations of triclosan with:

8- (2 '2 ' -Dimethyl -l'H-pyran-7-yl) coumarin;

[ (2 '2-Dimethyl-3-β-hydroxy-6-methylene) -1-β-cyclohexyl] -3 -methyl -pent-

2 -enyl-7 -oxycoumarin ;

[ (2'2,6-Trimethyl-2-oxo-bicyclo) -2, 2, 1-heptyl] l-β-3-methyl -pent-2-enyl-

7 -oxycoumarin, •

7-Cyclohexylmethoxycoumarin; and

Galbanic Acid provided compositions that were effective to reduce antimicrobial blocking resistance by at least 20 percent.

7-1,1 -Dimethylprop-2-ynyloxycoumarin 7-Methoxycoumarin

7-[3-Methyl-lbutenyloxy]coumarin

Example 4 - Pseudopeptides

This example summarizes the effectiveness of pseudopeptides on the prevention of Triclosan (IRGASAN DP300) antimicrobial resistance development in E. coli ATCC 9637. In this case, 5μl of lOmg/ml solution was embedded for each compound on a 6mm sterile disk. The diameter of the zone of inhibition was measured in mm after incubation for 5 days at 37°C. The number of resistant colonies within the zone of inhibition was recorded down as RC values. The results are shown Table 4-1.

TABLE 4-1

Strain E. coli ATCC 9637

As can be seen from Table 4-1 , the results show that only the combinations of triclosan with: N-tert-butoxycarbonyloxyaminopentan-N, N-diethylamide , • N-Benzoylalanyl-N, N-diethylamide , • N-tert-Butoxycarbonyloxyalanyl-N, N-diethylamide; N-tert-Butoxycarbonyloxyalanyl-N, N-morphilinoamide; N-tert-butoxycarbonyloxyalanyl-N-methyl-N-phenylamide; 4-Benzoylaminobenzoyl-N, -diethylamide ; 4-Benzyloxycarbonyloxybenzoyl-N, N-diethylamide ; 4-Toluenesulf onylaminobenzoyl-N, N-diethylamide ; 4-Benzoylaminobenzoyl -N, N-diethylamide ; 4-Benzyloxycarbonyloxybenzoyl -N, -diethylamide ; 5 -N-Benzylaminopentanoyl-N, N-diethylamide ; 5 -N-Cyclopentylaminopentanoyl-N, N-diethylamide; and 5 -N-Benzoylaminopentanoyl-N, N-diethylamide provided compositions that were effective to reduce antimicrobial blocking resistance by at least 20 percent .

N-tert-butoxycarbonyloxyamιnopentan-N,N-dιethylamιde N-benzyloxycarbonylalanyl-N,N-dιethylamιde

N-tert-Butoxycarbonyloxyalanyl-N,N-d.ethylam.de N-tert-butoxycarbonyloxyalanyl-N-methyl-N-

N-tert-Butoxycarbonyloxyalanyl-N,N-morphιhnoamιde 5-N-toluensefulfonylamιnopentanoyl-N,N-dιethylamιde

N-Benzylalanyl-N,N-dιethylamιde

BOC = tert/butoxycarbonyl Me = CH₃ Example 5 - Indazole Analogs

This example summaries the effectiveness of Indazole analogs on the prevention of Triclosan (IRGASAN DP300) antimicrobial resistance development in E. coli ATCC 9637. In this case, 5μl of lOmg/ml solution was embedded for each compound on a 6 mm sterile disk. The diameter of the zone of inhibition was measured in mm after incubation for 5 days at 37°C. The number of resistant colonies within the zone of inhibition was recorded down as RC values .

TABLE 5-1

Strain E. coli ATCC 9637

As can be seen from Table 5-1 , the results show that only the combinations of triclosan with: 1 -Methyl- 3 -benzyloxy- 5 -nitro- Iff- indazole; l-Methyl-3- [ (3-dimethylamino) propyloxy] -lff-indazole; 1 -Benzyl -3 -ethoxy-5 -ni tro-lff-indazole, • l-Methyl-3 -ethoxy-5 -benzoylamino-lff-indazole; 1 -Methyl- 3 -ethoxy-5-benzylaminocarboxylamino- lff-indazole; 1 -Methyl -3 -tosyloxy- 5 -ni tro-lff-indazole; 1 -Methyl -3 -bromo- 5 -nitro- lff-indazole, • 1 -Methyl -3 -benzyl oxy- 5 -amino- lff-indazole ; 1 -Phenyl -3 -hydroxy-5-ni tro-lff-indazole ; 1 -Benzyl -3 -benzyloxy- 5 -ni tro-lff-indazole; l-Methyl-3 -benzyloxy-5-nitro-lff-indazole; l-Methyl-3- [ ( 3-dimethylamino) propyloxy] -5-amino-lff-indazole ; 1 -Benzyl -3 -ethoxy-5 -amino-lff-indazole;

Benzydamine l-Methyl-3- [ (dimethylamino) propyloxy-lff-indazole ; Isoniazid or 4-pyridinecarboxylic acid hydrazide; l-methyl-3 -ethoxy-5-nitro-lff-Indazole ; and 1 -methyl -3 -ethoxy-5 -amino- lff-indazole provided compositions that were effective to reduce antimicrobial blocking resistance by at least 20 percent .

3-ethoxy-5-N-Toluene-sulfonyl-l/7-ιndazole l-benzyl-3-hydroxy-5-nιtro-l //-indazole

l-methyl-3-hydroxy-5-nιtro 1 /-indazole 1 -phenyl-3-hydroxy-5-nιtro 1 //-indazole

Me

Isoniazid or 4-pyπdιnecarboxyhc acid hydrazide l-methyl-3-ethoxy-5 amιnoben?yl-l //-indazole

3

-prenyl-4-hydroxy-cιnnamyl methyl ester l-methyl-3-hydroxy-5-amιno 1 //-indazole hydrochloπde

l-methyl-3-ethoxy-5-l//-ιnadazole 1 -methyl-3-ethoxy-5-amιno 1 H-Indazole

Et = CH₂CHι Me = CH₃ Ph = Phenyl Example 6- Isoglabrene Analogs

This example summarizes the effectiveness of flavanoids and isoflavanoids including isoflavenes, isoflavanes, isoflavanones and isoglabrene analogs on the prevention of Triclosan (IRGASAN DP300) antimicrobial resistance development in E. coli ATCC 9637. In this case, 5 μl of lOmg/ml solution was embedded for each compound on a 6 mm sterile disk. The diameter of the zone of inhibition was measured in mm after incubation for 5 days at 37°C. The number of resistant colonies within the zone of inhibition was recorded down as RC values .

TABLE 6-1

Strain E. coli ATCC 9637

As can be seen in Table 6-1, the results show that only the combinations of triclosan with: 4 ' -Methoxyisof lav-3-ene; 4 ' -Hydroxyisof lav-3 -ene; 2 ' -Methoxyisof lav-3 -ene; 7,2' -Dimethoxyisof lav-3 -ene; 2,4' -Dimethoxyisof lav-3 -ene ;

7-Methoxy-3- [5-methoxy-2 , 2-dimethyl-3 , 4-dihydro-2ff-l-benzopyran6- yl]2-ff-] -benzo-3-pyran; 2 ' O-Methylisoglabrene; Isoglabrene;

2 ' -O-Methyl-3 , 4, 3 "4" -tetrahydroisoglabrene; 3 " 4 " -Dihydroisoglabrene ;

2 ' 0-Methyl-7-demethyl-7-methanesulf onylisoglabrene; 7-Hydroxy-6-aceto-2 , 2-dimethylbenzo-lH-benzopyran; 5-Hydroxy-6-acetyl-2 , 2dimethyl-2ff-l-benzopyran;

5-Methoxy-6- [ (2-hyroxymethyl-5-mesyloxy) -2-phenyloxy] aceto-2,2- dimethylbenzo-lff-benzopyran; and 2-hydroxy-4-mesyloxybenzylacohol provided compositions that were effective to reduce antimicrobial blocking resistance by at least 20 percent.

2'4',7-Trimethoxyisoflav-3-ene 2'0-Methyl-7-demethyl-7-methanesulfonylisoglabrene

2-hydroxy-4-mesyloxybenzylacohol

2'4',-Dihydroxyisoflav-3-ene

6'O-Methylisoglabrene

Isoflav-3-ene

47-Dihydroxyisoflav-3-ene 47-Dimethoxyisoflav-3-ene Example 7- Antioxidants

This example summarizes the effectiveness of antioxidants on the prevention of Triclosan (IRGASAN DP300) antimicrobial resistance development in E. coli ATCC 9637. In this case, 5 μl of lOmg/ml solution was embedded for each compound on a 6 mm sterile disk. The diameter of the zone of inhibition was measured in mm after incubation for 5 days at 37°C. The number of resistant colonies within the zone of inhibition was recorded down as RC values .

TABLE 7 Strain E. coli ATCC 9637

As can be seen from Table 7, the results show that only the combinations of triclosan with: 6-hydroxy- 2 , 5, 7, 8-tetramethylchroman-2-carboxylic acid (TROLOX™), reduced glutathione, butylated hydroxyanisole (BHA) , butylated hydroxytoluene (BHT), (-) -Epigallocatechin Gallate (EGCG) , (-) -Gallocatechin Gallate (GCG), (-) -Epigallocatechin (EGC) , (-) -Epicatechin Gallate (ECG) and (-) -Epicatechin (EC) provided compositions that were effective to reduce antimicrobial blocking resistance by at least 20 percent.

It should be understood that the foregoing disclosure and description of the present invention are illustrative and explanatory thereof and various changes in the size, shape and materials as well as in the description of the preferred embodiment may be made without departing from the spirit of the invention.

Claims

What is claimed is:

1. A topical antimicrobial composition comprising:

(c) a topical antimicrobial agent; and

(d) an antimicrobial resistance blocking effective amount of at least one antimutagenic and/or antioxidant agent .

2. The composition of claim 1, wherein the antimicrobial agent is selected from the group consisting of nisin, bis- diguanides, chlorhexidine gluconate, chlorhexidine digluconate, chlorhexidine diacetate, chlorhexidine dihydrochloride, polyhexamethylene biguanide, benzalkonium chloride, benzethonium chloride, methylbenzethonium chloride, cetyl pyridinium chloride, triclosan, triclocarban, tribromsalani, amyltricresols, parachlorometaxylenol, phenol, silver, iodine, nonylphenoxypoly (ethyleneoxy) ethanoliodine, poloxameriodine complex, undecoylium chloride, iodine complex, bisquaternary ammonium compounds, polymeric quaternary ammonium compounds, alcohols, cationic peptides, organometallic antiseptics, alkyl pyridinium salts, essential oils, and combinations and derivatives thereof .

3. The composition of claim 2, wherein the antimicrobial agent is triclosan.

4. The composition of claim 2, wherein the antimicrobial agent is chlorhexidine.

5. The composition of claim 2, wherein the antimicrobial agent is benzalkonium chloride.

6. The composition of claim 2 , wherein the antimicrobial agent is benzethonium chloride.

7. The composition of claim 2, wherein the antimicrobial agent is triclocarban.

8. The composition of claim 2, wherein the antimicrobial agent is silver.

9. The composition of claim 2, wherein the antimicrobial agent is an alcohol selected from the group consisting of ethanol, propanol, butanol, pentanol, 2- methyl-1-butanol, hexanol, 2-methyl-l-pentanol, 3- methyl-1- pentanol, 2-ethyl-l-butanol, 3 , 5 , 5-trimethyl- 1-hexanol, heptanol, octanol, isooctyl alcohol, decanol, dodecanol, tridecanol, tetradecanol and mixtures thereof .

10. The composition of claim 2, wherein the antimutagenic or antioxidant agent is selected from the group consisting of substituted and unsubstituted pyrithione-containing compounds; coumarins; pseudopeptides; indazoles; antioxidants; flavanoids; isoflavanoids including isoflavenes, isoflavanes, isoflavanones and isoglabrene analogs; and mixtures thereof .

11. The composition of claim 10, wherein the substituted and unsubstituted pyrithione-containing compounds are monovalent or divalent metallic salts of pyrithione.

12. The composition of claim 11, wherein the pyrithione-containing compound is zinc or sodium pyrithione.

13. The composition of claim 12, wherein the pyrithione-containing compound is zinc pyrithione.

14. The composition of claim 12 or 13 wherein the antimicrobial agent is triclosan.

15. The composition of claim 10, wherein the coumarins are 8- (2 ' 2 ' -Dimethyl-1 Η-pyran-7-yl) coumarin ; [ (2 ' 2-Dimethyl-3-β-hydroxy-6-methylene) -1-β- cyclohexyl] -3-methyl-pent-2-enyl-7-oxycoumarin; [ (2 '2, 6- Trimethyl-2-oxo-bicyclo) -2,2, 1-heptyl] -l-β-3-methyl- pent-2-enyl-7-oxycoumarin; 7-Cyclohexylmethoxycoumarin ; and galbanic acid.

16. The composition of claim 15 , wherein the antimicrobial agent is triclosan.

17 . The composition of claim 10 , wherein the pseudopeptides are :

N-Tert-butoxycarbonyloxyaminopentan-N, -diethylamide ; N-Benzoylalanyl-N, N-diethylamide; N-tert-Butoxycarbonyloxyalanyl-N, -diethylamide ; N-tert-Butoxycarbonyloxyalanyl-N, N-morphilinoamide; N-Tert-butoxycarbonyloxyalanyl-N-methyl-N-phenylamide ; 4-Benzoylaminobenzoyl-N, N-diethylamide; 4-Benzyloxycarbonyloxybenzoyl-N, -diethylamide ; 4-Tol+uenesulf onylaminobenzoyl-N, N-diethylamide ;

4 -Benzoylaminobenzoyl-N, N-diethylamide; 4-Benzyloxycarbonyloxybenzoyl-N, N-diethylamide ; 5-N-Benzylaminopentanoyl-N, N-diethlamide ; 5-N-Cyclopentylaminopentanoyl-N, N-diethylamide ; and 5-N-Benzoylaminopentanoyl-N, N-diethylamide .

18 . The composition of claim 17 , wherein the antimicrobial agent is triclosan .

19. The composition of claim 10, wherein the indazoles are : l-Methyl-3 -benzyloxy- 5 -ni tro-lff-indazole ; l-Methyl-3- [ (3 -dimethylamino) propyloxy] -lff-indazole ; l-Benzyl-3 -ethoxy-5 -nitro- Iff- indazole, ^• l-Methyl-3 -ethoxy-5 -benzoylamino-lff-indazole ; l-Methyl-3 -ethoxy-5 -benzylaminocarboxylamino- Iff- indazole; 1 -Methyl -3 -tosyloxy- 5 -ni tro-lff-indazole, ^• l-Methyl-3 -bromo-5-ni tro-lff-indazole ; l-Methyl-3 -benzyloxy-5-amino-lff-indazole, ^• l-Phenyl-3-hydroxy-5-nitro-lff-indazole;

1 -Benzyl -3 -benzyl oxy- 5 -ni tro-lff-indazole ;

1 -Methyl -3 -benzyl oxy- 5 -ni tro-lff-indazole ; l-Methyl-3- [ (3-dimethylamino) propyloxy] -5-amino-lff- indazole;

1 -Benzyl -3 -ethoxy-5 -amino-lff- indazole; l-Methyl-3- [ ( dime thylamino) propyloxy- Iff- indazole;

Isoniazid or 4-pyridinecarboxylic acid hydrazide;

1 -methyl -3 -ethoxy-5 -ni tro-lff-indazole; and

1 -methyl -3 -ethoxy-5 -amino- Iff- Indazole.

20. The composition of claim 19, wherein the antimicrobial agent is triclosan.

21. The composition of claim 10, wherein the flavanoids and isof lavanoids .

22. The composition of claim 21, wherein the isof lavonoids are of substituted and unsubstituted isoflavanes, isoflavenes, isoflavones and isoglabrene analogs .

23. The composition of claim 21, wherein the isoglabrene analogs are:

4 ' -Methoxyisof lav-3 -ene; 4 ' -Hydroxyisof lav-3 -ene; 2 ' -Methoxyisof lav-3 -ene; 7,2' -Dimethoxyisof lav-3 -ene ; 2,4 ' -Dimethoxyisof lav-3-ene;

7-Methoxy-3 [5-methoxy-2 , 2-dimethyl-3 , 4-dihydro-2ff-l-benzopyran6- yl] 2-ff-] -benzo-3-pyran;

2 ' O-Methylisoglabrene;

Isoglabrene;

2 ' -O-Methyl-3 , 4, 3 "4" -tetrahydroisoglabrene;

3 "4 " -Dihydroisoglabrene;

2 ' 0-Methyl-7-demethyl-7-methanesulfonylisoglabrene;

7-Hydroxy-6-aceto-2, 2-dimethylbenzo-lH-benzopyran;

5-Hydroxy-6-acetyl-2 , 2-dimethyl-2iϊ-l-benzopyran;

5-Methoxy-6- [ (2-hyroxymethyl-5-mesyloxy) -2-phenyloxy]aceto-2,2- dimethylbenzo-lff-benzopyran; and

2-hydroxy-4-mesyloxybenzylacohol .

24. The composition of claim 23, wherein the antimicrobial agent is triclosan.

25. The composition of claim 10, wherein the antioxidants are 6-hydroxy-2 , 5 , 7 , 8-tetramethylchroman-2- carboxylic acid (TROLOX™) , reduced glutathione, butylated hydroxyanisole (BHA) , butylated hydroxy toluene

(BHT), (-) -Epigallocatechin Gallate (EGCG) , (-)-

Gallocatechin Gallate (GCG) , (-) -Epigallocatechin (EGC) , (-) -Epicatechin Gallate (ECG) and (-) -Epicatechin (EC) .

26. The composition of claim 25, wherein the antimicrobial agent is triclosan.

27. The composition of claims 1-10, wherein the relative ratio of topical antimicrobial agent to antimutagenic and/or antioxidant agent is in the range of from about 1 to about 0.1-50 based on weight and vice versa.

28. The composition of claims 1-10, wherein the antimicrobial agent and the antimutagenic and/or antioxidant agent are each present in a concentration amount ranging from about 0.0001 to about 50 mg/ml .

29. The composition of claim 28 where the antimutagenic and/or antioxidant agent is a pyrithione- containing compound present in a concentration amount ranging from about 0.1 to 5 μg/ml .

30. A method for inhibiting bacterial resistance comprising:

(b) applying the composition of (a) to a surface containing bacteria.