US 3928607 A
A number of water-soluble, one-to-one quaternary ammonium adducts of unsaturated organic halides and dihalides with hexamethylenetetramine are known as being useful as antimicrobial agents. It has now been discovered that a two-to-one quaternary ammonium adduct can be prepared from hexamethylenetetramine and 3,4-dichlorobutene-1; this two-to-one adduct is a more active, less toxic antimicrobial agent than the known one-to-one quaternary ammonium adducts.
Description (OCR text may contain errors)
tetl t afiQix Luloff et al.
ANTIBACTERIAL COMPOSITION AND METHOD EMPLOYING A CERTAIN HEXAMETl-IYLENETETRAMINE ADDUCT Inventors: Jerome S. Luloff, Bloomington,
Minn.; Albert L. Eilender, Flanders, NJ.
Cosan Chemical Corporation, Clifton, NJ.
Filed: Oct. 5, 1973 Appl. No.: 403,819
US. Cl. 424/249; 260/248.5; 106/15 AF; 424/70 Int. Cl. A01N 9/22 Field of Search 424/249; 260/248.5
References Cited UNITED STATES PATENTS l/l966 Wolf et al 424/249 Primary Examiner-V. D. Turner Attorney, Agent, or Firm-Pennie & Edmonds  ABSTRACT A number of water-soluble, one-to-one quaternary ammonium adducts of unsaturated organic halides and dihalides with hexamethylenetetramine are known as being useful as antimicrobial agents. It has now been discovered that a two-to-one quaternary ammonium adduct can be prepared from hexamethylenetetramine and 3,4-dichlorobutene-1; this two-to-one adduct is a more active, less toxic antimicrobial agent than the known one-to-one quaternary ammonium adducts.
2 Claims, No Drawings 1 ANTIBACTERIAL COMPOSITION AND METHOD EMPLOYING A CERTAIN HEXAMETHYLENETETRAMINE ADDUCT BACKGROUND OF THE INVENTION During 1915 and 1917 a number of papers were published by Dr. Walter Jacobs and coworkers at the Rockefeller Institute for Medical Research describing the general bactericidal character of quaternary ammonium salts of hexamethylenetetramine. The bactericidal activity of these compounds was primarily attributed to the presence of the hexamethylenetetramine nucleus; however, the degree or extent of this activity was said to be determined by the nature of the molecular groups adducted with the hexamethylenetetramine. The principal mode of action of these quaternary am monium salts of hexamethylenetetramine was said to be the controlled, regulated decomposition of the hexamethylenetetramine nucleus to liberate formaldehyde. The following group of papers by Jacobs and coworkers are of interest in this regard: Proc. Nat. Acad. Sci. USA, 1, 226 (1915); J. Biol. Chem., 20, 659 (1915); J. Biol. Chem, 21, 465 (1915); J. Exp. Med, 24, 563 (1916); and J. Exp. Med., 25, 363 (1917). These Jacobs papers, however, do not describe any quaternary ammonium adducts of hexamethylenetetramine with unsaturated acyclic dihalides.
In US. Pat. No. 3,228,829 Wolf and a coworker at the Dow Chemical Company have more recently described as antimicrobial agents a group of one-to-one quaternary ammonium adducts of hexamethylenetetramine with unsaturated halides and dihalides. Among the useful unsaturated halides and dihalides described are the dihaloalkenes and haloalkynes, specifically CH2 CH 9 1,3-dichloropropene.
In French Pat. No. 1,363,240 Frank and coworkers at Farbenfabriken Bayer even more recently described as antibacterial agents a group of one-to-one quaternary ammonium adducts of hexamethylenetetramine with oz,w-dihalogenated hydrocarbons. Among the group of useful a,w-dihalogenated hydrocarbons described is 1,4-dichlorobutene-2.
We have now discovered that a two-to-one quater- CH CH Cl Cl-I=Cl-l2 CHCH BASIC PARAMETERS OF THE INVENTION A. Preparation of the Product When hexamethylenetetramine and the dihalides of Wolf and Frank react, only one of the halogens is displaced by hexamethylenetetramine to form a quaternary ammonium salt. The second halogen, however, is
-unreactive and, therefore, only a one-to'one quaternary ammonium adduct is formed. On the contrary,
when hexamethylenetetramine and 3,4-dichlorobutene-l react, both chlorines are displaced by the hexamethylenetetramine so that a two-to-one quaternary ammonium adduct is formed.
Contrary to the reaction of the Wolf and Frank dihalides, it is believed that the reaction of this invention between hexamethylenetetramine and 3,4-dichlorobutene-l proceeds by a rearrangement mechanism which results in the presence during the reaction of two reactive sites on the dihalide and thus allows for the prepa ration of the two-to-one rather than the one-to-one quaternary ammonium adduct. The reaction is believed to occur in the following fashion, with the rearrangement being accomplished by the allylic displacement of the 3-chlorine and concomitant shift of the double bond:
Regardless of the actual mechanism of the reaction, the analytical data available on the product clearly indicates that the twoto-one quaternary ammonium adduct is formed. Particularly, the analysis for percent ionic chlorine and percent nitrogen in the final product indicate that the two-to-one quaternary ammonium adduct is obtained according to this invention. Moreover, nuclear magnetic resonance and infrared data indicate the product is symmetrical about the double dichlorobutene-l to form the two-to-one quaternary ammonium adduct of this invention should advantageously be carried out in an inert organic solvent in which the reactants are substantially soluble and the product is substantially insoluble. Suitable solvents are hydrocarbon and chlorinated hydrocarbon solvents 1 such as chloroform and perchloroethylene.
The reaction should be conducted between 0C. and a maximum upper limit of about 130C., the decomposition point of the two-to-one quaternary ammonium adduct. There are no necessary pressure conditions for the reaction, and the reaction can thus be carried out at sub, super or normal atmospheric pressure. However, use of super atmospheric pressure reduces substantially the reaction time.
Consequently, the only two requisite reaction parameters are that 1) the reaction be carried out in an inert organic solvent in which the product is substantially insoluble and (2) the reaction temperature be maintained at less than about 130C. to avoid decomposition of the product.
the process of this invention.
Table I on pages 5 represents a comparison of analytical data relating to the organic-ionic chlorine content of products obtained according to this invention and products obtained by varying the conditions of example 1 of the Frank patent. The data in Table I confirm that the product of this invention is a two-to-one rather than one-to-one quaternary ammonium adduct.
with 3 ,4- 30 Additionally, determinations of the percent nitrogen in the product of this invention confirms that the twoto-one quaternary ammonium adduct is formed. Table II is a comparison of the percentages of nitrogen calculated and obtained for the product of this invention and the oneto-one quaternary ammonium adduct formed according to example 1 of the Frank patent.
TABLE II Nitrogen Nitrogen Calculated Obtained Repetition of Example 1 of the Frank patent 21.1% 22.6% Procedure according to this invention using 3,4dich1orobutene-1 27.6% 27.8%
Nuclear magnetic resonance and infrared analysis confirm that the product of this invention is the twoto-one quaternary ammonium adduct, presumably prepared according to the rearrangement mechanism out lined hereinabove. These data indicate l) the absence of monosubstituted ethylene or vinyl groups and (2) high symmetry abut the double bond; both of these structural aspects confirm that the final product is completely or almost completely made up of the symmetrical two-to-one adduct. However, the presence of insignificant amounts of a one-to-one adduct cannot be conclusively ruled out by these techniques.
As a result of the elemental analysis for organic chlorine, ionic chlorine and nitrogen and the nuclear magnetic resonance and infrared analysis, it has been determined that the product made according to this invention is almost completely, if not completely, made up of the symmetric two-to-one quaternary ammonium adduct of 3,4-dich1orobutene-1 with hexamethylenetetramine, i.e. 1,1-(but-2-enylene) bis(3,5,7-triaza-1- azoniaadamantane chloride).
C. Antimicrobial Activity of the Product The two-to-one quaternary ammonium adduct of hexamethylenetetramine and 3,4-dich1oro butene-l is a more active antimicrobial agent than either of the oneto-one quaternary ammonium adducts of Wolf and TABLE I Organic Organic Ionic Ionic Chlorine Chlorine Chlorine Chlorine Calculated Calculated Calculated Calculated Product and Total Total On Total On Total Organic On Total On Total Ionic Preparation Chlorine Chlorine Chlorine Chlorine Chlorine Chlorine Chlorine Chlorine Technique Calculated Obtained Calculated Obtained Obtained Calculated Obtained Obtained Repetition of Example 1 of Frank patent 26.6% 24.7% 13.3% 12.4% 10.8% 13.3% 12.4% 13.9% Repetition of Example 1 of Frank patent with large excess of l,4-dich1orobutene-2 26.6% 25.9% 13.3% 13.0% 11.8% 13.3% 13.0% 14.1% Repetition of Example 1 of Frank patent using extended reflux and large excess of 1,4 dich1orobutene-2 26.6% 25.7% 13.3% 12.9% 11.7% 13.3% 12.9% 14.0% Procedure according to this invention using 3,4-dich1orobutcne-I 17.5% 17.7% 0 O 0.4% 17.5% 17.7% 17.3% Procedure according to this invention using 3,4-dich1orobutene-l 17.5% 17.1% 0 0 2.1% 17.5% 17.1% 15.0% Procedure according to this invention using 3,4-dich1orobutane-1 17.5% 17.1% 0 O 1.0% 17.5% 17.1% 16.1% Procedure according to this invention using 3,4dich1orobutene-l 17.5% 16.1% o 0 oa /l 17.5% 16.1% 15.9%
Frank. The higher activity is due to the fact that there are two hexamethylenetetramine nuclei in the product of this invention, both of which upon decomposition liberate formaldehyde. Consequently, the same amount by weight of the two-toone quaternary ammonium adduct of this invention is more active as an antimicrobial agent than the same amount by weight of the one to-one adducts. of Frank and .Wolf. In this sense more active means that assuming that (l) the amount of quaternary ammonium adduct used and (2) the rate of formaldehyde release are the same, the antimicrobial activity of the two-to-one quaternary ammonium adduct of this invention will be more persistent as the two-to-one adduct will release formaldehyde for a significantly longer period of time. Consequently the same amount of the two-to-one quaternary ammonium adduct of this invention is capable of controlling larger numbers of microorganisms.
Additionally, the two-to-one adduct formed from hexamethylenetetramine and 3,4-dichlorobutene-l according to this invention is less toxic than either the Frank or Wolf one-to-one adducts. For instance, data comparing the toxicological character of the two-. to-one adduct of this invention to that of both the one to-one adduct of example I of Frank and the one-toone adduct prepared from hexamethylenetetramine and l,3-dichloropropene according to Wolf (available commercially as Dowicil 100 and 200) indicate the two-to-one adduct to have a significantly superior toxicological profile.
Table III shows these comparative data for both acute dermal and acute oral LD tests. Both the acute oral LD and acute dermal LD tests were run in accordance with standard, accepted protocols.
Data available from Dow Chemical on Dowicil l00. "Data available from Dow Chemical on Dowicil 200.
From the data in Table III, it is apparent that the two-to-one adduct of this invention is markedly superior from a toxilogical standpoint than the prior art oneto-one adducts.
D7 The Antimicrobial Uses of the Product The two-to-one quaternary ammonium adduct of this invention is a water-soluble, non-metallic, nonphenolic antimicrobial agent effective against a broad spectrum of gram positive and gram negative microorganisms. The two-to-one quaternary ammonium adduct of this invention is particularly useful as an antimicrobial agent in a wide variety of water-based systems such as latex paints, resin emulsions, joint cement, adhesives, dispersed pigments and dyes, and other similar aqueous compositions.
The two-to-one quaternary ammonium adduct of this invention is useful in the preparation and storage of systems susceptible to deterioration by microorganisms such as the common forms of bacteria. The exact concentration of the adduct during use is dependent upon a number of factors including duration of storage, temperature of storage, etc. However, concentrations in the range of 0.05 to 0.20% based upon the weight of the composition have been found to be generally effective. It should be noted, however, that the minimal inhibitory concentrations vis-a-vis the common bacteria are much less than this suggested range for use under commercial conditions.
SPECIFIC EMBODIMENTS OF THE INVENTION The following Examples are non-limitative embodiments of the invention and are included merely as specific examplification of the invention.
EXAMPLE 1 To a 1 liter flask equipped with stirrer, thermometer and reflux condenser is charged:
chloroform 600 mls hexamethylenetetramine grams -3,4 dichlorobutcne-l 125 grams sodium carbonate l0 grams The reaction mass is refluxed for 30 hours and then cooled to' room temperature and'filtered. The filter cake is washed three times with a total of 750 mls of chloroform and then allowed to air dry. The theoretical calculated yield is 111 grams while the actual weight obtained -is 108 grams.
EXAMPLE 2 To a 2 liter stirring autoclave the following charge is added:
chloroform 1,020 mls hexamethylenetetramine I 19 grams butylated hydroxy toluene 0184 grams 3,4 diehlorobutene-l 212.5 grams sodium carbonate 8.5 grams The reaction is maintained at C for a period of 18 hours with a maximum generated pressure of 12 psi. At the end of the reaction period, the mixture is cooled and filtered. The filter cake is then washed with 350 ml increments of chloroform three times and then placed in a vacuum oven at 40 to constant weight. The theoretical calculated yield is 180.5 grams while the actual yield obtained is 166 grams.
EXAMPLE 3 To a 1 liter flask equipped with stirrer, thermometer and reflux condenser is charged:
perchloroethylene 600 mls 3,4 dichlorobutene-l grams hexamethylenetramine 70 grams sodium carbonate 10 grams The reaction is heated at 75C for 25 hours. Product mix is cooled and then filtered. Filter cake is washed 4 times with 250 ml increments of perchloroethylene and then air dried. The theoretical calculated yield is l l 1 grams and the actual yield obtained is 104 grams.
EXAMPLE 4 The product of this invention had demonstrated especially high antibacterial performance in the preset ya tion of latex and emulsion based paints. The followlhg is a description of a notable paint formulation.
7 To a 1 gallon stainless steel beaker, equipped with a sheer type agitator, the following ingredients are charged in sequence:
In a separate 100 ml beaker, the following premix is prepared:
9 grams 24 grams hydroxyethylccllulosc coupling solvent This premix is then added to the above with agitation such that excessive air is not incorporated. To the above prepared combination is added 1,40l grams of a vinyl acrylic resin emulsion and mixed for a period of 10 minutes.
The resulting paint prepared above was examined for antibacterial properties by inoculating a portion with approximately 10 organisms per ml Pseudomonas aeruginosa. Paint samples returned to sterility within 24 hours of inoculation and maintained the same degree of preservation for periods in excess of 9 months, at which time the testing was discontinued. The addition of test compound preserved the paint system against attack from microbial contamination and thereby maintained the viscosity, stability and performance characteristics of the paint.
EXAMPLE 5 The following is a formulation for an oil-water hair dressing emulsion:
Part A is heated to C and Part B is heated to 0C.
The test compound (Part C) is dissolved in Part B and-- the resulting solution added to Part A with good agitation. The mixture was perfumed at 45C and agitated until cold.
The composition prepared above was tested for microbial stability for a period of 6 months and after 6 month at 35C. Inoculation with bacterial cultures at this point showed the material still resistant to bacterial contamination.
1. An aqueous composition normally susceptible to deterioration by bacteria containing the two-to-one quaternary ammonium adduct of hexamethylenetetramine and 3,4-dichlorobutene-l in an amount effective to exert antibacterial activity, said adduct having the structural formula:
9 6) N N-CH2CH=CHCH2-N N 2. A method of controlling bacteria comprising contacting said bacteria with the two-to-one quaternary ammonium adduct of hexamethylenetetramine and 3,4-dichlorobutene-l in an amount effective to exert antibacterial activity, said adduct having the structural formula:
9 o CH CH=CHCH N