CA2069453A1 - Biocide composition - Google Patents

Abstract

ABSTRACT
BIOCIDE COMPOSITION
A composition which comprises a cyclic thiohydroxamic acid derivative, including a metal complex thereof, together with a biocide material which is i) an agent affecting membrane permeability, ii) an isothiazolinone, or iii) an aldehyde. The cyclic thiohydroxamic acid derivative may be a zinc salt or complex of 3-hydroxy-4-methylthizol-2(3H)-thione. The agent affecting membrane permeability may be a phenol such as o-phenylphenol or may be a biguanide, including a polymeric biguanide, or a quaternary ammonium compound. The isothiazolinone may be 1,2-benzisothiazoline-3-one or 5-chloro-2-methylisothiazolin-3-one. The aldehyde may be formaldehyde or glutaraldehyde. The composition has biocidal activity.

Description

2~9~5~

BIOCID~ COMPOSITION

The present invention relates to compositions which are useful as industrial biocides.

Industrial biocides are useful to prevent industrial spoilage, in particular that caused by bacteria and fungi. Materials which can be used as industrial biocides have antimicrobial properties and particularly have antifungal and antibacterial properties. Such materials are useful in the preservation of paints, latices, adhesives, leather, wood, metal working fluids and cooling water.

Many compounds have been disclosed which have anti-microbial activity and some of these compounds are commercially available and are used as industrial biocides. ~owever, whilst these materials can provide use~ul protection, a need remains for materials having improved characteristics.

In our European Patent Application Publication No 249328, we disclose that certain cyclic compounds have useful antimicrobial properties and can be used as biocides. We have now found that when used with other compounds having antimicrobial properties a beneficial effect can be achieved.

According to the present invention there is provided a composltion which comprises A) at least one compound of the formula (I) / \
B N - O~
\ I (I) C = S

D

i ~ .

:~
`~ ~
.

2~6~3 -2- S 3~34 or a salt or complex thereof; and B~ at least one other biologically active compound which is (i) an agent affecting membrane pe~neability;
(ii) an isothiazolinone or an isothiazolothione; or (iii) an aldehyde;
wherein A is a nitrogen or carbon atom, which may be substituted7 B and D are, independently, oxygen or sulphur or a nitrogen or carbon atom which may be substituted; or A and/or B and/or B and/or D may be part of a ring syste~;
R is hydrogen, a hydrocarbyl group, a substituted h~drocarbyl group, an acyl group, a substituted acyl group or a group -COORl; and Rl is a hydrocarbyl group with the proviso that ~ and D ~re not both sulphur or both oxygen.

The bonds between the groups A and B and between the groups B and D may be single or double bond~ but it will be appreciated that there can be only one such double bond in the ring.

The group A, and optionally one or both of groups B and D
can be a group -CtR2)2-; a group -CR2~; a group ~ C-NR2; a group -NR - or a group N-; where R is a hydrogen atom, a hydrocarbyl group, a substituted hydrocarbyl group or two groups R2, together with the carbon atom, or carbon atoms, to which they are attached form a ring.

The groups A, B and D can form part of a Eurther r~ng system but generally not more than two of the groups A, B ancl D form part o a further ring system. The further ring system is typically a hydrocarbon ring system containing five or six carbon atoms, for example a cyclopentene, cyclohexane, cyclohexene, cyclohexadiene or benzene ring. The further ring system, if present, typically contains one or both of the groups A and B. I~ only the group A
forms part of a ring system, this may be a cyclohexane ring of the type - - . -.

. , . ~ . . :
, , , ~ " , - . ... .

4 ~ ~
-3_ S 35834 CH ~ ~ CH2 \ /

/
CH2 --- C~2 where the group A is the carbon atom with the ~uo free valences, which are linked to the group -NOR- and B respectively. If both A
and B form part of a ring system, the further ring is then fused to S the azolethione rin~ system; for example as in 3-hydroxy-4,5,6,7-tetrahydrobenzothiazol-2~3H)-thione.

I~ many of the compounds used in the biocide compositions of the present invention, the groups ~1 B and/or D are not part of a fur~her ring system. Thus, if A, B and/or D is a carbon atom, or substituted carbon atom, it may be, inter alia9 a group -CH=, C(C2H5) ~ -C(C6~5)~, -C(C6~4Cl)~ -C(C~3)2 or C=N~. It will be appreciated that in the foregoing, the group R2 is a hydrogen atom, a methyl, ethyl, phenyl or chlorophenyl group. The group R2 typically is a hydrogen atom, a lower alkyl group, that is one containing up to five carbon atoms, an aryl group or a subs~ituted alkyl or substituted aryl group. If the group R2 iY a substituted group, each substituent ls a hydrocarbonoxy group, a hydro~arbonthio group, an acyl (that is a hydrocarboncarbonyl) group, an es~er (that is an acyloxy) group, a halogen ~tom, a nitrile group or a hydroxy group.

Preferred compounds are those in which the groups A and B
are both optiotlally substituted carbon atoms and the group D is a sulphur atom or an optionally substituted nitrogen atom. l'he groups A and B are preferably linked through a double bond as in the group -CR2~CR2, in which the groups R2 may be the same or different. It is preferred that D is a sulphur atom.

: . , .: ' . ~ :

2~9~3 _4_ S 3583 The group R may be a hydrogen atom, an acyl group such as benzoyl or acetyl, or an alkoxycarbonyl group such as an ethoxycarbonyl group. If the group R is a substituted group the substituent may be as disclosed for the group R2 or may be a substituent which contains a further ring system of general formula I, the two ring systems being linked through the group R, for example as in the glu~aryl bis ester of the formula:

OCoc~2c~2cH2coo I
N N
C~3 ~ \ ~ ~r CH3 ?
/c=Ss~c ~11 S S

Component (A) of the composition may be a salt or complex of the compound of general fonmula ~. The salt or co~plex may be one which contains any metal. The metal may be a transition metal, for example a metal of group VIII, IB or IIB of the Periodic Table. Such metals include iron, copper and zinc, particularly such metals in their m~ximum possible valency s~ate.
:
All references herein to the Periodic Table are to the Pe~iodic Table according to MendeleePf, as set out on the inside rear cover of UGeneral and Inorganic Chemistry" by J.R.Partingon, Second Edition publlshed by MacMillan and Co.Limited, I.ondon.

For convenience hereafter, the compounds of the general formula I, and salts and complexes thereof wlll be referred to simply as "Compound I".

A wide range of Compounds I can be used as component (A) in ~he compositions of the present invention. The Compounds I have antl-microbial activity agalns~ a wide range of micro-organisms.

., ~
.
.', ~.

.~ . : ~ , , ~ . .
.
- - . ' ::

~069~3 _5_ S 35834 Compounds I which can be used in the compositions of the present inventioD include:
3-hydroxy-4-methylthiazol-2(3H)-thione, 3-benzoyloxy-4-methylthiazol-2(3H)-thione, 3-hydroxy-4-phenylthiazol-2t3H)-thione, 3-hydroxy-4,5,6,7-tetrahydrobenzothiazol-2t3H)-thione, 3-acetoxy-4-methylthiazol-2(3H)-thione, the glutaryl bis-ester of 3-hydroxy-4-methylthiazol-2(3H)-thione, 5,5-dimethyl-1-hydroxy-4-imino-3-phenylimid~æolidine-2-thione, 1-hydroxy-4-inino-3-phenyl-2-thiono-1,3-diaza~piro[4.5]decan~, l-hydroxy-5-methyl-4-phenylimidazoline-2-thio~e, 3-ethoxycarbonyloxy_4-methylthiazol-2(3H)-thione, 4,5-dimethyl-3-hydroxythiazol-2(3EI)-thione, ~s,5-dimethyl-3-acetoxythiazol-2t3H)-thione, 4-ethyl-3-hydroxy-5-methylthiazol-2(3H)-thione, 4-ethyl-3-acetoxy-5-methylthiazol-2(3H)thione, 4-(4-chlorophenyl)-3-hydroxythiazol-2(3H)-thione, 3-hydroxy-S-methyl-4-phenylthiazol-2(3~)-thione, 3-ace~oxy-4-phenylthiazol-2(3H)-thione, and th~ metal complexes and salts thereof. The metal complexes and salts thereof includ~ ferric, cup~ic and zinc complexes and salts such as the zinc complex of 3-hydroxy-4-methylthiazol-2(3H)-thione, thP
ferric complex of 3-hydroxy-4-methylthiazol-2(3H)-thione, the cupric complex of l-hydroxy-4-imino-3-phenyl-2-thion-1,3-diazaspiro~4.S]decane, the cupric complex of 4,5-dimethyl-3-hydroxythiazol-2(3H)~thione, the zinc complex of 4,5-dimethyl-3-hyclroxythiazol-2(3H)-thione, and the zinc complex of 4-ethyl-3-hydroxy-5-methylthiazol-2(3H)-thione.

We have obtained particularly useful results when component (A) is the zinc complex or salt of 3-hydroxy-4-methylthiazol-2(3H) -thione.

' 2~9~L~3 Component (B) (i) of the composition i5 an agsnt affecting membrane permeability. Action on general membrane permeability is discussed in ~Pharmaceutical Microbiology" edited by W.B.Hugo and J.B.Russell, published in 1977 by Blackwell, Chapter 11, specifically at pages 202 and 204. The d~scussion indicates that treatment of bacterial cells with certain substances causes a leakage of a group of characteristic chemical species. Po~assium ion is indicated to the first substance to appear when such damage occurs. Materials which cause such an effect can be used as component (B) (i) of the composition of the present invention. Component (B) (i) may be a phenol and o-phenylphenol has been found to give a useful effect.

Cationic disinfectants can be effective as an agent affectinK membrane permeability. By disinfectan~ we mean a material which is able to destroy pathogenic micro-organisms but not necessarily resistant spores as discussed on page 701 of "Medical Microbiology", 11th edition (1965) by R.Cruickshank, published by E
and S Livingstone Ltd. Effective cationic disinfectants for use in the compositions of the present invention include blguanides and quaternary ammonium compounds.

Biguanides which may be used in the compositions of the present invention contain at least one biguanide unit of the formula (II) _ -NC-C-NH C-NH- (II) NH NH
.
Typically the biguanide contains at least two units of the formula (II) which are linked by a bridging group which contains at least one methylene group. The bridging group may include a polymethylene chain which may optionally be interrupted by hetero atoms such as oxygen, sulphur or nitrogen. The bridging group may include one or more cyclic nuclei which may be saturated or unsaturated. It is generally preferred that the bridging group is such that there are at leas~ three, and especially at least four, .
''~: ' ' ' ' ' ~

-- . . ::

. . ~ .

2~9~5~

-7 S 3~34 carbon atoms directly interposed between two adjacent units of the fonmula (II). In general it is preferred that there are not more than ten carbon atoms, especially not more than eight carbon atoms, interposed betw~en two adjacent units of the formula (II).

The biguanide units May be terminated by any suitable group which may be a hydrocarbyl or substituted hydrocarbyl group or which may be an amine group or an amine hydrochloride group or by a group -NH-C-NH-CN
Il NH
If the terminating group is a hydrocarbyl group this may be an alkyl, cycloalkyl or aryl group or may be an alkyl, cycloalkyl or ,aryl group or may be a combination thereof as ln an aralkyl group. If the .erminating group is a substituted hydrocarbyl group, the substltuent can be any substituent which does not have an undesirable adverse effect on the microbial activity of the biguanide compound and typically is a hydrocarbonoxy group, a hydrocarboncarbonyl (than is an acyl) group, an ester (that is an acyloxy) group, a halogen atom or a nitrile group and there may be more than one substituent, for example more than one halogen atom.

A suitable biguanide is a material which contains two units of ~he formula (II) and in which the the units are linked by a ; polymethylene group, particularly a hexamethylene group. The terminating groups may be 4-chlorophenyl groups, for example as in the compound oE formula (III):-H H El H H H
~1 1 1 1 i I ~
Cl ~ ~ N-C-N-C-N-~CH~)6-N-C-N-C-N~ ~ Cl (III) ~ ~v NH NH NH NH

The compound of formula (II) is available as a chlorhexidine salt.

::
.~ ~

2~9~3 The biguanide may alternatively be a polymeric biguanide, for example a linear pol~neric biguanide which has a recurring polymer unit represented by the formula X-NH-C-NH-C-NH-Y-NH-C-NH-C-NH-Il 11 11 11 NH NH NH NH

wherein X and Y may be the same or different and represent bridgin8 groups in which together the total number of carbon atoms directly interposed bet~een the pairs of nitrogen atoms linked by X and Y is at least 9 and not more than 17.

The bridging groups X and Y may consist of polymethylene chains, optionally interrupted by hetero atoms, for example, oxygen, sulphur or nitrogen. X and Y may also incorporate cyclic nuclei ~hich may be saturated or unsaturated, in which case the number of carbon ato~s directly interposed between the pairs of nitrogen atoms linked by X and Y is taken as including that segment of the cyclic group, or groups, which is the shortest. Thus, the number of carbon a~oms directly lnterposed between the nitrogen atoms in the group ~ .
-NH-CH2-O CH2 N~l is 4 and not 8.

The preferred polymeric biguanide for use in the present inventiorl is poly(hexamethylene biguanide), in ~hich X and Y both represent the -(CH2)6- group.

Polymeric biguanides may be prepared by the reaction of a bisdicyanidiamide having the fonmula ,:
.

20~9~

CN-NH-C-NH-X-NH C-NH-CN
Il 1i NH NN

with a diamine H2N-Y-NH2, wherein X and Y have the meanings defined previously herein;

5or by reaction between a diamine salt of dicyanimide having the formuLa :~ + +
(H3N-X-NH3)(N(CN)2)2 with a diamine ~2N-Y-NH2 wherein X and Y have the meanings defined ~; previously herein. These methods of preparation are d~scribed in UK
lOPatent Specifications Nos. 702,268 and 1152243 respectively, and any of the polymeric biguanides described therein may be used as component (B)(i) in the composition of the present invention.

The polymer chains are terminated either by an amino ~; hydrochloride grnup or by an -NH-C-NH-CN group, and the terminating 11 , NH
`
groups may be the same or different on each polymer chain.

Typically, the polymerlc biguanides are obtained as mixtures of polymers in which the polymer chains are of different lengths, the number of individual biguanide units, i.e.

NH NH

and -Y-NH-C-NH-C-NH-, NH NH

. . ~ ., .

2~9~3 10- S 35~34 together being from 3 to about 80.

In the case of the preferred poly(hexa~lethylene biguanide) having the formula (IV) (CH2)6-NH-C-NH-C-NH 1--NH N~ (IV) rl the value of n is in the range from 6 to 15, the average molecular weight of the polymer mixture being from about 1100 to about 3300.

The biguanides are used as salts with suitable inorganic or organic acids, for example as the hydrochloride salts or the acetate or gluconate.

The component (B)(i) may be a quaternary a~monium salt of the general formula ~V):-[NR3R4RSR6] E ~V) where R3 is a hydrocarbyl group or substituted hydrocarbyl group;
R4, RS and R6 may be the same or different and are a hydrogen atom or a hydrocarbyl group or a substituted hydrocarbyl group; and E is an anion havlng a valency and being in an a~ount: to giYe a neutral compound;
wherein R3, R4, R5 and R6 tog~ther contain at least six aliphatic carbon atoms or two or more of R4, R5 and R6, together with the nitrogen atom to which they are attached, form one or more heterocyclic rings.

Preferred quaternary ammonium salts are those in ~hich R3 is a group containing at least six allphatic carbon atoms. If any of the groups R3, R4, RS or R6 is a substltuted hydrocarbyl group the subs~ituent may be a hydrocarbonoxy group, an acyl group, an acyloxy group, a nltrile group, a hydroxy group or a halogen atom.

"
-2~6~3 The hydrocarbyl group may be an alkyl, cycloalkyl or aryl group or a mixture of such groups as in an alkaryl group. The quaternary ammonium compound may one in wh:ich not more than two the groups R3, R4, R5 and R6 are alkyl groups containing at least six carbon atoms. In a compound of the foregoing type the remaining groups are preferably hydrogen, lower alkyl (as previously defined herein) or a group con~aining an aromatic ring such as a benzyl group. Alternatively, the residual groups may fonm a heterocyclic ring, for example a pyridine ring or a nitrogen containing adamantane structure.

The anion E is preferably a mono-valent anion such as a halide anion, a bisulphate (HSO4) anion or an alkylsu].phate anion.

Quaternary a~monium compounds which may be used as component (B)(i) in the compogition of the present invention include diethyldodecylbenzyl ammonium chloride;
dimethyloctadecyl-(dimethylbenzyl)am~onium chloride;
dimethyldidecylammonium chloride; dimethyldidodecylammonium chloride;
; trimethyl-tetradecylam~tonium chloride; benzyldimethyl(C12-C18 alkyl)a~ttonium chloride; dichlorobenzyldimethyldodecylammonium chloride; hexadecylpyridiniu~ chloride; hexadecylpyridinium bromide;
hexadecyltrimethylammonium bromide; dodecylpyridinium chloride; !' dodecylpyridinium bisulphate;
benzyldodecyl-bistbeta-hydroxyethyl)~ttmonium chloride;
dodecylbenzyltrimethylammonium chloride; benzyldimethyltC12-C18 alkyl~ ammonium chloride; benzyldimethyl (C12-C16 alkyl) ammoniu~
chloride; trimethyl ~C12-C16 alkyl) ammonium bromide;
dodecyldimethylethyl ammonium ethylsulphate;
dodecyldimethyl (l-naphthylmethyl)ammonium chloride;
hexadecyldinethylbenzyl al~nonium chloride; dodecyldimethylbenzyl an~tonium chloride and l-t3-chloroallyl)-3,5,7-triaza-1-azonia-adamantane chloride. The quaternary ammonium salt may be benzyldimethyl(C12-C16)alkyl ammonium chloride.

2~694L~3 -12- S 35~34 Component (B)(ii) of the composition is an lsothiazolinone or an isothia~olothione.

The isothia~olinone or isothiazolothione derivstive which is component (B)(ii) of the composition is typically a compound of the general fonmula:

R8 \ ¦ ¦ :

/ C

; ¦¦ N - R7 C

. R9 S

wherein:
2 is an oxygen or sulphur atom;
R7 is a hydrogen atom, a substituted or unsubstituted hydrocarbyl:group, a substituted or unsubstituted hydrocarbylthio group, a substituted or unsubstituted hydrocarbyloxy group, a carbamoyI group or a cation;
R8 i8 a hydrogen atom, a halogen atom, a cyano group, or a ~ubstituted or unsubstituted hydrocarbyl group;
R9 is a hydrogen atom, a halogen atom, a cyano group, or a substituted or unsubstituted hydrocarbyl group; or R~ and R9, together with the carbon atoms to which they are attached, form a five- or six-membered ring, which may optionally be substituted.

;: :
:, -2~9~3 Preferably component (B)(ii) is at least one isothiazolinone derivative, ~hat is a compound in which Z is an oxygen atom. If the groups R7, R8 and R9 are, or contain, substituted hydrocarbyl groups, the substituents may be as disclosed for the group R2 and are typically halogen, alkoxy or alkylthio, particularly those in which the alkyl groups contain l to 4 carbon atoms. If R7 is a carbamoyl group, this of th~ general type -CONHR10 where R10 is a hydrogen atom or a hydrocarbyl group, which may be substituted. It is generally preferred that the group R7 is a hydrogen atom or a lower alkyl group (as previously defined herein).
R7 is especially hydrogen or a methyl group.

R~ and Rg ~ay, together with the carbon atoms to which they are attached, form a flve- or six-membered ring, which may be substituted, the substituents typically being halogen, alkyl, alkoxy, or alkylthio gro~ps. The ring thus ob~ained may contain a heteroatom, for example a nitrogen atom but in general R8 and R9 fon~ a hydrocarbon ring such as a benzane, cyclopentene or cyclohexene ring.
Alternatively, R8 and R9 are separate groups and one or both o R8 and R9 can be a hydrogen atom. It is generally preferred that at least one of R8 and R9 is other than a hydrogen atom and is, particul~rly, a halogen atom, for example chlorine or a lower alkyl ; group.

Isothia~olinone compound (Z is oxygen) which can be used as component (B)(ii) of the mixture include 2-methylisothiazolin-3-one (R7 is methyl, R8 and R9 are both hydrogen);

5-chloro-2-methylisothiazolin-3-one (~7 ls methyl, R8 is hydrogen and R is chlorine); mi~turqs of the foregoing two compounds;
4,5-dichloro-2-methylisothiazolin-3-one ~R7 is methyl and R8 and R9 are both chlorine); 1,2-benzisothiazolin-3-one (R7 is hydrogen and R8 and R9, together with the carbon atoms to which they are attached, ~ form a benzene ring); 4,5-trimethylene-4-isothiazolln-3-one (R7 is ; hydrogen and R8 and R9 togethcr with the csrbon atoms to which ~hey are attached, ~orm a cyclopentene ring); and 2-methyl-4,5-trimethylene-4-isothiazolin-3-one (R7 is methyl and R~
and R9 together with the carbon atoms to which they are attached, form a cyclopentene ring).

.
: . .

~ S 3583 In component tB)(ii). if R7 is a cation this may he a cation having a valency of more than one but is particularly a monovalent cation such as an alkali metal, an amlne or quaternary ammonium cation.

Component (B)(iii) of the composition is an aldehyde.
Suitable aldehydes are fonmaldehyde and glutersldehyde.

Useful results have been achieved when component (B) is o-phenylphenol, chlorhexidine hydrochloride salt, poly(hexamethylene biguanide) hydrochloride salt, (C12-C16) al~yldimethyLbenz~lammonium chloride, a mixture of quaternary a~monium chlorides including (Cl2-Cl6)alkyldimethylbenzyl, octyldecyldimethyl, dioctyldimethyl and didecyl dimethyl ammonium chloride, 5-chloro-2-methylisothia~olin-3-one, 1,2-benzisothiazolin-3-one, 2-methyl-4,5-trinethylene-~-isothiazolin-3-one, Eormaldehyde or glutaraldehyde.
:, Useful compositions in accordance with the present invention comprise A) the zi~c complex or salt of 3-hydro~y-4-methylthiazol-2(3H)-thione; and B) o-phenylphenol,chlorhexidine hydrochloride salt, poly(hexamethylene biguAnide) hydrochloride salt, (Cl~-Cl6) alkyldimethylbenzylammonium chloride, a mlxture of quaternary ammonium chlorides including (C12-C16)alkyldimethylbenzYl, octyldecyldimethyl, dioctyldimethyl and didecyl dimethyl ammonium chloride, 5-chloro-2-methylisothiazolin-3-one;
1,2-benzisothiazolin-3-one;
2-methyl-4,5-trlmethylene-4-isothiazolin-3-one, :Eormaldehyde or glutaraldehyde.

The relative proportions of components (A) and (B) are dependent on the particular componen~s, especially component ~B).
The antimicrobial activity of materials which can be used in the compositions of the present invention varies quite widely, this being particularly so in respect oE materials which can be used as component (B), some of which are effective at a concentration of less 2 ~ 5 3 than lo ppm by ~rei~ht whilst other materials may be used in amounts as high as 0.1% by weight to achieve an effect. Hence, depending on the particular combination of (A) and (B), the relative proportions thereof, by weight, may be from lO0:1 to 1:50.

The composition of the present invention may be prepared by mixing components (A) and (B) together using any suitable technique depending on the physical state of the components and their solubility. Thus, A and B may both be solids and be mixed together using any suitable solids mixing technique. Alternatively, A and B
may be both be liquids and be mutually miscible. A and B may both be soluble in the same solvent and be mixed together us solutions in said solvent. A further alternative is to suspend one component which is a solid in a liquid medium which is the other component or is a solution o the other component in a liquid medit~n in which only lS one of A and B is soluble.

The mixtures of the present invention provide improved anti-microbial ac~ivity allowing lower concentrat~ons of component (A) to be used.

The compositions of the present invention provide good wet state preservation making the compositions advantageous for use as a cutting fluid preservative and also in cooling water applications.
Wood and leather preservation is another advantageous field of application of the compositions. The compositions o~ the present invention can also be incorporated into paint7 as paint film ftmgicide and many of the compositions can be used without the addition oP a further active component.

The compositions oE the present invention may consist only of components (A) and (B). However, typically the composition comprises the components (A) and (B) as a solution, suspension or emulsion in a suitable liquid medit~n such as water. The composition may comprise a suspension or emulslon of the components ~A) and (B), or a solution thereof, in a liquid medium in ~hich o~e, or both, of the components is inso1uble.

, . ~
,: ~

.; . - . .~ ~:

2 0 ~ 3 -16- S 35~34 The composition may be incorporated into a medium to be protected against micro-organisms using any suitable ~ixing technique. Thc composition is incorporated into the medium to be protected in an amount to provide from 0.00002 to 5% by weight of the composition of (A) and (B) relative to the total compositiont more preferably from 0.00005 to lZ by weight of (A) and (B). It will be appreciated that the quantity of the composition of (A) and (B) required will be dependent on various factors such as the medium to be protected, the micro-organisms against which protection is desired and the extent of protection required.

If the composition is being used to preserve a solid substrate such as leather or wood, the composition may be applied directly to the substrate or may be incorporated into a coating composition such as a paint, varnish or lacquer which is then applied to the substrate. Alternatively, the solid material may be impregnated with the composition of the present invention.

The compositions of the present invention can be used for the treatment o$ various media to inhibit the growth of micro-organisms.

Thus, as a further aspect of the present invention there i9 pro~ided a method for inhibiting ~he growth of micro-organisms on, or in, a medium which comprise~ treating the medium with a composition o ~A) and (B) as hereinbefore defined.

The composition can be used in conditions in which micro-organisms grow and cause problems such as, for example, in AqUeOus environments including cooling water systems, paper mill liquors, metal working fluid~s geological drilling lubricants, polymer emulsions, and emulsion paints. The composition can also be used to impregnate solid materials such as wood or leather or can be coated onto the surfaces thereof directly or incorporated into a paint, varnish or lacquer.

- .

~, - . - : , 20~9~3 As a Eurther aspect the present invention provides a material which is susceptible to attack by micro-organisms and which cont~ins a composition of (A) and (B) in sn amount sufficient to at least reduce attack by micro-organisms.

Further aspects of the present invention are described in the following illustrative examples.

In the fcllowing exarnples, the products obtained were subjected to microbiocidal evaluation. The microbiological testing was effected, as follows:

~_ , Soil was taken from a depth o up to 100 mm, the top 25 mm being discarded. To each gramme of soll (wet weight) was added 2 cm3 of sterile saline (0.85% w/v sodium chlorlde). The resulting mixture was stirred for 30 minutes and allowed to settle for five minutes.
The liquid was separated from large particulate matter using a syphon.

20 cm3 of the resulting liquid was added to malt broth to give a final volume of 1 dm3.

, Evaluati~n .~ L~

lO cm3 of the soil/broth mixture prepared as described were placed into 2S cm3 bottles. To the contents of each bottle were added quantities of one or more bioc:Ldes to gi~e an ll x 11 array containing different relative proportions of biocide or biocide mixture. The bottles were vortex mixed for 1-3 seconds and incubated for one weelc at 25C without agitation.

::~ ` , , ~, .. , ; ' ' ' ~

2~6~3 -18- S 35~34 The bottles were inspected and any containing visible bacterial or ungal growth were recorded and the contents thereof discarded. 10 mm3 aliquot samples were removed from the remalning bottles and these were placed on malt agar plates, allowed to dry and incubated at 25C for two to three days. The presence or absence of growth was recorded and the bottles providing the aliquots giving growth were discarded.

Immediately after removing the 10 mm3 aliquot samples, 200 mm portions of freshly prepared Soil Organism Sample were added to the remaining bottles. After a fur~her week, lO mm3 aliquot samples were removed, placed on malt agar plates and evaluated for growth.

The procedure described was con~inued until 3 portions, each of 200 mm3, of freshly prepared Soil Organism Sample had been added to the bottles containing the initial soillbroth mixture.

Lsamples l to lO

A number of compo~nds and mixtures were tested in the manner described. The concentrations of biocide, or biocide mixture, which prevented any viable organisms gi~ing rise to growth in the fourth 10 mm3 aliquot placed on a malt a~ar plate were noted as providing effective control of the soil organis~s.

The results obtained were used to plot the variation of FLC
with the relative proportions of the biocide components. These results are set out in accompanying Figures l to 11, wherein Figure 1 is a plot of the variation of E'LC using ZHMT ~the zinc complex or salt of 3-hydroxy-4-methylthiazol-2(3H)-thione) or 1,2-benzisothiazolin-3 one (BLT) or mixtures thereof;

Figure 2 is a plot of the variation of FLC using ZE~T or formaldehyde or mixtures thereof;

-- .

~6~3 -l9- S 3583 Figure 3 is a plot of the variation of FLC using ZHMT or 1,2-dibromo-2,4-dicyanstobutane (T38) or mixtures thereof;

Figure 4 is a plot of the variati.on of FLC using Z~T or glutaraldehyde or mixtures thereof;

Figure 5 is a plot of the variation of FLC using ZHMT or chlorhexidine or mixtures thereof;

Figure 6 is a plot of the variation of PLC using ZHMT or 2-methyl-4,5-trimethylene-4-isothiazolin-3-one (MTI) or mi.~tures thereof;

Figure 7 is a plot of the variation of FLC using ZE~T or 5-chloro-2 methylisothiazolin-3-one (CIT~ or mixtures thereoE;

Figure 8 is a plot of ~he variation of FLC using Z~MT or ; (G12-C16)alkyldimethyl-ben~yl~mmonium chloride (VCL) or mixtures thereof;

Figure 9 i9 a plot of the variation of FLC using ZHMT or o-phenylphenol or mixtures thereof;
~ .
Pigure lO is a plot of the variation of PLC using ZEMT or a mixture of quaternary ammonium chlorides (available as Bardac 205M
(205) from Lonza Inc) or mixtures thereof;

Pigure ll is a plot of the variation of FLC using Z}~T or a po].~(hexamethylenebiguanide) hydrochloride having an average molecular weight in the range 1850 to 3300 (PHB) or mixtures of ZHMT
and PHB.

In Figures 1 to 11, the concentration of the biocides are given as a proportion of the fractional lethal concentration (FLC) where PLC = Concentration of compound in mixture which controls ~rowth Concentration of compound alone which controls growth.

, 2~69~3 -20~ S 35834 In Figures 1 to 11, the dotted straight line represents the relationshlp if an additive effect is achieved. The 8011d line represents the relationship as d0termlned experimentally. In Figure 3, using a mi~ture which is not in accordance with the present invention, the dotted and soLid lines coincide showing that this mixture gives an additive effect.

It should be noted that not all o~ the biocides used with Z~MT were effective under the test conditions described even at the highest concentration tested and with these materi~ls a value of one on the axis of the graph represents a nu~erical extrapolation to a point where the compound alone gives control.

From the plots in Figures l to 11, it is possible to determine the minimum value of the FLC of the mixtures used and the proportions of the two components which are required to attain this minim~lm value. This information is se~ out in the following Table together with results using further mixtures which ar~ not represented in Figures 1 to 11 including mixtures, not in accordance with the present invention, which did not give a minimum value of FLC.

`

2 ~ 3 TABLE
. _ _ ~
¦ Example I Z~MT ta) I Blocide _ ¦ Total I or Comp ¦ Amount I Type ( c ) I Amount ¦ FLC
¦Ex. ¦ (ppm) (b) ¦ (c) ¦ (ppm) (b) ¦ (d) __ l _ .. _ l l _ l _ ¦ A ¦ 50 I NIL ¦0 ¦ 1.0 ¦ B ¦ 0 ¦ BIT ¦63 ¦ 1.0 ¦ 1 ¦ 20 I BIT I22 1 0.75 ¦ C ¦ 0 ¦ FA ¦1600 ¦ 1.0 ¦ 2 ¦12.5 I FA ¦400 ¦ 0.5 ¦ D ¦12.5 ¦ GA ¦2000 ¦ 1.0~ 1 ¦ 3 ¦ 15 I GA ¦1200 1 0.75 ~ ¦
¦ E ¦ 0.0 ¦ CH ¦6000 ¦ 1.0 12.5 I C~ 11200 1 0.45 F ¦12.5 ¦ MTI I 60.0 ¦ 1.0 ¦ 5 ¦17.5 ¦ MTI ¦ 36 ¦ 0.8 ¦ G I Q.0 I CIT I 2.4 ¦ 1.0 1 6 117.5 I CIT ¦ 1~2 ¦ 0.85 ¦ H ¦ 0.0 j VCL ¦ 350 ¦ 1.0 1 7 ¦ 15 ¦ VCL ¦ 140 ¦ 0.7 I I ¦ 0.0 1 OPP ¦ 300 ¦ 1.0 1 8 120.0 1 OPP I 105 1 0.75 ; I J I 0 1 205 1 125 1 1.0 9 112.5 1 205 1 37.5 1 0.55 I K ¦12.5 ¦ PHB I 200 ¦ 1.0+
¦10 ¦ 15 ¦ P~B I 120 1 0.75+ ¦
¦ L ¦ 0 ¦ T38 ¦ 100 ¦ 1.0++ ¦
¦ M ¦ 30 ¦ T38 1 70 ¦ 1.3++ ¦

¦ N ¦ ltO ¦ T38 ¦ 10 ¦ 0.9+
¦ 0 ¦ 0.0 I BR0 ¦ 49.2 ¦ 1.0 ¦ P ¦62.5 ¦ BR0 ¦ 49.2 ¦ 2.25 I Q ¦ 75 ¦ BR0 ¦ 24.6 ¦ 2.0 ¦ R ¦ 0 ¦ DS ¦1000 ¦ 1++
¦ S ¦62.5 ¦ DS ¦1000 1 2.25t+ ¦
I r 1 75 I DS I500 1 2.0 ¦ U ¦ 0 ¦ DA l1000 ¦ 1.0 I V 187.5 ¦ DA ¦800 ¦ 2.55-~
~~ ~ . I

~0~5~

Notes to Table a) ZEMT is the zinc complex or salt of 3-hydroxy-4-methylthiazol-2(3H)-thione and can be prepared as described in Example 2 of EP-A-249328.

(b) Amoun~ is the concentration of ~he active componen~ in parts per million w/v of the liquid mixture.

tc) BIT is 1,2-benzisothiazoline-3-one FA is formaldehyde GA is glutaraldehyde CH is chlorhexidine MTI is 2-methyl-4,5-trimethylene-4-isothiazolin-3-one CIT is 5-chloro-2-methylisothiazolin-3-one ;~ VCL is (C12-C16)alkyldimethylbenzyl ammoni~ chloride OPP ls o-phenylphenol 205 is a mixture of quaternary ammonium chlorides available as Bardac 205M from Lonza Inc.
PHB is poly(hexamethylene biguanide)hydrochloride having an average molecular weight in the range 1850 to 3300 T38 is 1,2-dibromo-Z,4-dicyanatobutane BR0 is 5-bromo-5-nitro-1,3-dioxan DS is Z,3,5,6-tetrachloro-4-methylsulphonylpyridine DA is 3-(2-chlorobenzyl)-2,2-dimethyl-4-(triazol-1-yl)butan-3-ol (d) PLC is the fractional lethal concentration and i9 the concentration which prevents growth i~ ~he "Evaluation of microbial activity" test as described herein and for any compound i8 given by the relation~hip:-FLC - Concentxation of compound in mixture wh~ch prevents ~rowth Concentration of compound alone which prevents growth .

:~ :

2~69~3 + At the maximum level used, thsse compounds alone did not prevent growth under the test conditions described and were effective to prevent growth only in combination with a quantity of ZXMT. With these compounds, FLC is calculated as follows:-(Concn. of com~ound in mixture preventin~ ~rowth) x [l-~FLC of Z~MT
Highest concn of compound used at highest concn of other compo~:md) ]

Hence, if the highest concentration of a co~pound which prevents growth is 1000 ppm in the presence of 10 ppm of Z~MT, the ~: relationship is (Concn of compound in mixture prev nt n~ ~rowth) x (1-10) that is 0.8 (Concentration of compound in mixture preventl~rowth) .
++ At the maximum level used, these compound alone did not prevent ~rowth under the ~est conditions described and were ; effective to preven~ growth only in combination with a quantity of ZHMT but did not give a minimum value of FLCo With these compounds, PLC is determined using the highest concentration of the particular compound used, for example a compound alone does not give control at 1600 ppm but in a mixture with ZHMT control i8 achieved with 40 ppm of ZHMT
and 1~00 ppm of the compound, the FLC for this mixture is 40 ~ 1200 = 0.8 -~ 0.75 ~ 1.55 .

Claims (9)

1. A composition which comprises (A) at least one compound of the formula (I) (I) or a salt or complex thereof; and (B) at least one other biologically active compound which is (i) an agent effecting membrane permeability;
(ii) an isothiazolinone or an isothiazolothione; or (iii) an aldehyde;, wherein A is a nitrogen or carbon atom, which may be substituted;
B and D are, independently, oxygen or sulphur or a nitrogen or carbon atom which may be substituted; or A and/or B and/or B and/or D may be part of a ring system;
R is hydrogen, a hydrocarbyl, group, a substituted hydrocarbyl group, an acyl group, a substituted acyl group or a group -COOR1; and R1 is a hydrocarbyl group with the proviso that B and D are not both sulphur or both oxygen.

2. A composition as claimed in claim 1 wherein component (A) is a compound in which A, and optionally one or both of the groups B
and D are a group -C(R2)2-, a group -CR2=, a group C=NR2, a group -NR2 or a group -N=, wherein R2 is a hydrogen atom or an alkyl group containing up to five carbon atoms

3. A composition as claimed in either claim 1 or claim 2 wherein component (A) is the zinc salt or complex of 3-hydroxy-4-methylthiazol-2(3H)-thione.

4. A composition as claimed in any one of claims 1 to 3 wherein component (B)(i) is a phenol.

5. A composition as claimed in any one of claims 1 to 3 wherein component (B)(i) is a cationic disinfectant.

6. A composition as claimed in claim S wherein component (B)(i) is a biguanide or a quaternary ammonium compound.

7. A composition as claimed in claim 6 wherein component (B)(i) is a chlorhexidine salt or a poly(hexamethylene biguanide) having an average molecular weight of from about 1100 to about 3300.

8. A composition as claimed in claim 6 wherein component (B)(i) is a quaternary ammonium salt of the general formula (IV) [NR3R4R5R6] E (IV) where R3 is a hydrocarbyl group or substituted hydrocarbyl group;
R4, R5 and R6 may be the same or different and are a hydrogen atom, a hydrocarbyl group or a substituted hydrocarbyl group; and E is an anion having a valency, and being in an amount, to ; give a neutral compound;
wherein R3, R4, R5 and R6 together contain at least six aliphatic carbon atoms or two or more of R4, R5 and R6, together with the nitrogen atom to which they are attached, form one or more heterocyclic rings.

9. A composition as claimed in claim 8 wherein component (B)(i) is benzyl dimethyl (C12-C16) alkyl ammonium chloride.

,. .
,. :

: :

lo. A composition as claimed in either claim 1 or claim 2 wherein component (B)(ii) is a compound of the general formula wherein Z is an oxygen or sulphur atom;
R7 is a hydrogen atom a substituted or unsubstituted hydrocarbyl group, a substituted or unsubstituted hydrocarbylthio group, a substituted or unsubstituted hydrocarbyloxy group, a carbamoyl group or a cation;
R8 is a hydrogen atom, a halogen atom, a cyano group or a substituted or unsubstituted hydrocarbyl group;
R9 is a hydrogen atom, a halogen atom a cyano group, or a substituted or unsubstituted hydrocarbyl group; or R8 and R9 together with the carbon atoms to which they are attached form a five-or six-membered ring, which may optionally be substituted.

11. A composition as claimed in claim 10 wherein component (B)(ii) is a compound in which Z is an oxygen atom and R7 is a hydrogen atom or an alkyl group containing up to five carbon atoms.

12. A composition as claimed in either claim 10 or claim 11 wherein component (B)(ii) is 2-methylisothiazolin-3-one;
5-chloro-2-methylisothiazolin-3-one;
4,4-clichlore-2-methylisothiazolin-3-one; 1,2-benzisothiazolin-3-one;
4,5-trimethylene-4-isothiazolin-3-one or 2-methyl-4,5-trimethylcne-4-isothiazoline-3-one.

13. A composition as claimed in either claim 1 or claim 2 wherein component (B)(iii) is formaldehyde or gluteraldehyde.