NO142259B - AMINES FOR USE AS ANTIMICROBIAL AND / OR ALGEOUS INHIBITORS - Google Patents

Description

The present invention relates to a new group of compounds of substituted polyamines which are useful as algae inhibitors and as broad-spectrum antimicrobial agents, particularly against bacteria that cause plant diseases. The new compounds according to the invention have the structural formula: where A, which is the same or different, is cyclohexyl or cyclohexenyl with the formula:

where each R is either hydrogen or lower alkyl,

each R 1 , which is the same or different, is alkylene of 1-4 carbon atoms; and

Z is - (CH 2 ) 3 NH (CH 2 ) 3 NH 2 ; - (CH 2 ) NH(CI 2 ) 2 NH(CH 2 ) 2 NH 2 ;

- (CH2)3NCH3 (CH2)3NII2; -CH2CHOHCH2NH2; -(CH^NH^

-(CH2)2NH2 or -CH2"CHOH-NH2

and acid addition salts thereof.

The utility of the compounds according to the invention is generally their broad-spectrum antibacterial and antifungal properties. Particularly useful is their activity against bacteria and fungi which are responsible for crippling the growth and even destruction of many types of useful plants, and against those which cause the breakdown or deterioration of many types of materials. This includes paper, leather, textiles, aqueous preparations such as latex paints, adhesives, resins, pigment dispersions and oleoresin coatings whose films are particularly sensitive to the degrading action of fungi. The large economic losses that occur in paper manufacturing caused by the accumulation of bacterial and fungal mucus in various parts of the water system can be eliminated to a significant extent by using the compounds described here. In agriculture, a serious problem faced when growing cotton, beans, maize and other useful crops is the yield loss per acres due to the action of soil-borne fungi on the seeds and on the roots of the young plants. Combating or eliminating these losses can be achieved by using the compounds described here as soil disinfectants according to the invention. They can also be used on foliage and trees to combat bacterial and fungal diseases.

They are particularly useful as an agent active against bacterial diseases of fruit such as "fireblight" (caused by

Erwinia amylovora). The best agent currently useful is streptomycin, which is not only expensive, but as it is an antibiotic useful in human medicine, its use in agriculture is considered by some as a source of streptomycin-resistant species of

pathogens.

.,• The compounds according to the invention are preferably prepared by the following reaction sequence:

where A and Z are as indicated above, HX is a monovalent or polyvalent organic or inorganic acid, where sufficient HX is used to protonate at least one amino group of compound I.

The preparation of polyamine I involves the straightforward Schiff base reaction of the appropriate ketone IV and the appropriate amine V.

Since amine V has two primary amino groups, it can be either symmetrical or unsymmetrical. An amine V which is a symmetrical amine forms a single Schiff base VI. This is because no matter which primary amino group of amine V reacts with ketone IV,

the same product is formed. However, when amine V is unsymmetrical,

two products can be formed. One is Schiff base VI. Both prod-

ukter falls within the scope of the definition given for Schiff-

base VI. When two Schiff bases are formed, they can be separated, if desired,

by the usual and well-known separation methods, i.e. distillation and the like.

As an alternative to obtain a mixture of Schiff-

bases VI, the reaction can be carried out stepwise. For example, 1,2-diaminoethane can be transferred to a Schiff base with 1,5-di-(4-isopropylcyclohexyl)-3-pentanone, catalytically reduced, then the ring nitrogen is selectively cyanoethylated with acrylonitrile-', followed by catalytic hydrogenation to give [ 1,5-di-(4-isopropylcyclohexyl)-. 3-pentylj-1,4,8-triazaoctane.

To prepare Schiff base VI, ketone IV and amine V are dissolved

in a suitable inert solvent, e.g. toluene, and heated to reflux until reaction is substantially complete. Usually 5 - 20 hours are sufficient to remove the water by azeotropic distillation. The solvent is then removed under reduced pressure, and the residue comprising the Schiff base VI is dissolved in an inert solvent, preferably an alkanol, such as ethanol or isopropanol.

After dissolution, the Schiff base VI is catalytically reduced or

chemical.

If the reduction occurs catalytically, a possibly unsaturated carbon-to-carbon bond in A will also be reduced or hydrogenated, as well as the carbon-to-nitrogen bond of the Schiff base VI. In such catalytic reductions, hydrogen saturates an alkanol solution of Schiff base VI using stirring in the presence of the usual hydrogenation catalysts, such as transition metals and their reducible oxides. Particularly effective catalysts are the noble metals and their oxides. A particularly preferred catalyst is platinum oxide. In general, the hydrogenation reaction is carried out in a manner that is well known. Small particles, e.g. 100 - 300 mesh, of catalyst is mixed with the Schiff base, and excess amine in alcohol and placed in a closed system which is placed under a pressure of 3 - 5 atmospheres of hydrogen gas, preferably at room temperature, and usually at such pressures and a temperature from 15° to 45°C. At higher temperatures, the pressure preferably does not need to exceed 15 atm. After the reaction is complete, the pressure is released and the catalyst is separated from the reaction mixture by filtration. The filtrate containing cyclohexyl-polyamine I is then further purified by usual methods. Preferably, any solvent present is removed under reduced pressure, the residue is then dissolved in a water-immiscible solvent, washed with water, followed by a further wash with a saturated, aqueous, inorganic salt solution. After drying, the dissolution medium is removed by evaporation under reduced pressure, which gives the cyclohexylpolyamine I usually as an oil. The cyclohexyl polyamine can then be redissolved in lower alkanols, mixtures of lower alkanols and water, diethyl ether, dioxane and then neutralized with an acid, e.g. hydrogen chloride or neutralized directly with aqueous acids.

Acid addition salts are then isolated, if desired, by sifting,

evaporation or other commonly used methods.

Suitable anions X for the salt in (a) include anions derived from inorganic acids as well as those from organic acids such as halide, i.e. chloride, bromide, or iodide, or sulfate, nitrate, bisulfate, phosphate, acetate, propionate, maleate, succinate, laurate, palmitate, oleate, stearate, ascorbate, gluconate, citrate, carbonate, bicarbonate, benzoate, salicylate, pamoate , phthalate, furoate, picolinate, dodecylbenzene sulfonate, lauryl ether sulfonate, nicotinate, etc. In general, any anion derived from an acid is suitable and satisfactory when the polyamine salt anion X , e.g. chloride, is replaced with other anions, by well-known anion-exchange methods.

In the preparation of cyclohexenyl polyamines, i.e. the product I where the olefinic opening in ring A is retained, a selective chemical rather than a catalytic reduction is used to reduce Schiff base VI to product I.

In this chemical reduction method, the ketone IV is reacted with the appropriate amine as before, but the Schiff base VI dissolved in an inert alkanol or ether-type solvent is reacted with a chemical reducing agent such as sodium borohydride or lithium aluminum hydride.

Although as little as 1 equivalent of the chemical reducing agent can be used successfully, more satisfactory results are obtained if at least a 2 molar excess of, and preferably at least a 2.5 molar excess of, the chemical reducing agent is used. After any initial reaction has subsided, the reaction mixture of Schiff base VI and the reducing agent can be heated under reflux for an hour or two, then cooled to room temperature and then evaporated under vacuum. The residue obtained is further purified as by treatment with mineral acid or inorganic base as described for polyamines I, and the salt can then be formed as previously described.

The cyclohexyl and cyclohexenyl ketones IV are readily prepared, and two alternative methods are indicated below.

(A) The condensation with acids

This method involves the following reaction scheme:

Acylating decarboxylation of acids VII is used by heating the acid at elevated temperatures either with transition metals, preferably iron, transition metal oxides, alkaline earth metal oxides, with polyphosphoric acid or with boron trifluoride. Most conveniently, the acylating reaction is achieved by passing acid vapors over catalysts such as heated thorium oxide airgel.

Condensation-decarboxylation of an acid is the preferred method for the preparation of ketone IV when the two A-R^ groups are the same, as a mixture of products is obtained when several different acids are combined in a reaction. The preferred reaction involves mixing carboxylic acid VII with reduced iron powder and stirring in an inert atmosphere at 195° - 200°C for 1 - 6 hours to form an iron salt.

Preferably, the carboxylic acid VII and the iron are stirred under an inert atmosphere of nitrogen for at least 2 hours at 195° - 200°C.

After 2 hours, the temperature is suitably increased to 290° - 310°C, and stirring is continued for at least a 3-hour period, 4 hours being usually sufficient. The reaction mixture is allowed to cool, and then extracted with a suitable inert solvent such as diethyl ether and filtered. The solvent extracts are evaporated under reduced pressure. The remaining liquid is distilled under vacuum to isolate the ketone IV.

The carboxylic acids VII used above are prepared by various well-known methods. A particularly useful method is the addition of a cyclohexene to an aliphatic acid anhydride.

In this method, a mixture of cyclohexene and a catalytic amount, e.g. 0.2 - 0.3 mol for each mol of cyclohexene, of a free radical-forming catalyst, such as di-t-butyl peroxide, dropwise over 3 - 5 hours to a 5 - 15 molar excess of refluxing aliphatic acid anhydride. After the addition is complete, the reaction mixture is heated under reflux for 5 - 10 hours, evaporated under reduced pressure, and the liquid residue is mixed with aqueous sodium hydroxide and stirred while heating on a steam bath for approx. 2 - 5 hours. The cooled, alkaline solution is then extracted with ether, the ether layer is discarded, and the aqueous solution is made acidic, and then extracted well with ether. The combined ether extracts are washed with water, dried over anhydrous sodium sulfate and evaporated under reduced pressure. The remaining liquid or solid substance is distilled under vacuum to obtain a corresponding carboxylic acid, VII.

Other carboxylic acids are easily obtainable, e.g. by the Diels-Alder reaction of a diene and alkyl-substituted diene with various unsaturated aliphatic compounds or carboxylic acids, which will be discussed in more detail later. (B) Condensation of a Grignard reagent and a nitphil Dicyclohexy1-, dicyclohexenyl or cyclohexy1-cyclohexenyl alkanones can be obtained according to the following reaction scheme:

where A or R of each reactant may be the same or different, and are as previously stated.

This general procedure utilizes the reaction of a Grignard reagent prepared from a chlorine- or bromine-substituted cyclohexane or cyclohexene derivative with a cyano-substituted cyclohexane or cyclohexene derivative. The formed disubstituted iminoalkane salt complex is hydrolyzed with aqueous mineral acid to the corresponding ketone.

The Grignard reagent is obtained by reacting the halide with magnesium meta 11 , usually in the form of shavings or powder, and can be catalyzed with very small concentrations of iodine or methyl iodide. Solvents that are useful include diethyl ether, dibutyl ether, tetrahydrofuran, dioxane and benzene. Usually, gentle heating is sufficient to initiate the reaction, and the halide is gradually added to the metal-solvent mixture. After the final addition, the disappearance of practically all magnesium meta 11 shows that the reaction has ended. A small excess of halide is used, and moisture must be excluded; a nitrogen atmosphere is beneficial. The Grignard reagent is then added to the nitrile, which has been previously dissolved in two or three times its volume of solvent, over a period of 15 minutes to 1 hour at ambient temperature. The reaction mixture can then be heated to reflux to ensure complete reaction. In general, a small excess of Grignard reagent is used in relation to nitrile. From 1 to 10 hours of reflux is sufficient for complete transfer. The formed imine salt is preferably split into the ketone with aqueous mineral acids such as hydrochloric acid, sulfuric acid and phosphoric acid. The ketones are water-insoluble and can be extracted with water-immiscible solvents. Purification is preferably carried out by fractional distillation under reduced pressure. It is possible to use the crude ketone reaction mixture for the alkylation of polyamines as the reaction by-products are usually alcohols or hydrocarbons and do not react with amines. The reacting halides, if present in the crude product, should be removed prior to the ketone-amine alkylation process.

The concentrations of Grignard reagent and nitrile can be varied over wide ranges to ensure good yields in the process.

The halide and cyano, as well as carboxylic acid derivatives of cyclohexanes and cyclohexenes are commonly available. When the necessary carboxyl, cyano or halogen derivatives as used here are not readily available, they can be obtained by using known methods, e.g. using the Diels-Alder synthesis:

where D is R, -R-^COOH, -R^-Br or -R^CN and where R1 ,

R and X are as above indicated. When D is R and each R is alkyl, the cyclohexene formed can be reacted with an aliphatic acid anhydride as previously described. When D is R^COOH, R^CN or

R-^-Br, the condensation can take place as indicated in production examples A and B above. Of course, A-R^COOH can be treated by standard methods with a phosphorus chloride, e.g. phosphorus pentachloride,

to form A-R^-COCl.

When R is independently either hydrogen or alkyl of 1-6 carbon atoms, Darzen's synthesis [Compt. Rend., 150, 707 (1910)] is used: where , R and A are as indicated above. Likewise, Blaise-Marie's synthesis route can be used jBull. Soc. Chim. [4] 7> 215 (1910) and Compt. Clear. 145, 73 (1907) j:

where A and are as stated above.

When the ketone IV is obtained, it can then be reacted with a polyamine V which is triethylenetetramine, 3,3'-methylimino-bis-(propylamine), dipropylenetriamine, spermidine, ethylenediamine, trimethylenediamine, 1,3-diamino-2-hydroxypropane and 1 ,2-diamino-1-hydroxyethane.

The compounds described here are excellent broad-spectrum antimicrobial agents that are particularly effective against Gram-positives

and gram-negative bacteria, particularly the troublesome gram-negatives of the genus Pseudomonas at aqueous concentrations of 1.0 to 100 ppm. Examples of susceptible species include e.g. Staphylococcus aureus, Streptococcus pyogenes, Bordetella bronchiseptica, Pasteurella multocida, Escherichia coli, Salmonella typhimurium, S. pullorum, Klebsiella pneumoniae, Aerobacter aerogenes, Pseudomonas aeruginosa, Desulfovibrio desulfuricans, Bacillus mycoides, fungi such as Aspergillus niger and Chaetomium globosum. When used, these compounds can be applied neat or used in diluted form. Satisfactory diluents include any inert material which does not destroy the antimicrobial activity and in particular liquid preparations comprising aqueous dispersions, solutions and emulsions. Solid diluents include talc, cornstarch, aluminum oxide and diatomaceous earth. The antimicrobial agents of the invention can also be applied to materials such as natural fibers including paper, cotton, wool and synthetic fibers such as "Nylon", polypropylene, as well as to inanimate surfaces including hard surfaces such as wood, glass, metal, ceramic, rubber, plastic, and porous surfaces such as concrete, leather etc.

The polyamines according to the invention are particularly useful for suppressing the growth of aerobic and anaerobic bacteria in fluids used in cutting and grinding operations, such as metalworking, and oil well drilling mud or secondary oil recovery water and brines. Anaerobic microorganisms such as the sulphate reducer, Desulvo-vibrio desulfuricans, are inhibited at 0.1 - 10 ppm concentration of these polyamines. Suppression of these bacteria eliminates hydrogen sulphide formation and corrosion on equipment, plugging of oil-bearing sands, bad odors and other harmful effects. These compounds are also useful in the preservation against biodegradation of other aqueous systems such as aqueous emulsions and dispersions, paints or coatings, pigment suspensions, adhesives and the like, where the multiplication of microorganisms can cause colloid - breakdown, pH shift, bad odour, corrosive substances, loss of viscosity and other undesirable effects.

A particularly useful application of the compounds according to the invention is to impart sanitary properties to textiles, both woven and non-woven, washable or disposable, which are to be used e.g. such as nappies, surgical masks, caps, gowns, towels and curtains, covers for hospital furniture and instrument wrappings, aseptic face pads and sanitary napkins and toilet paper. In this application, the compounds of formula I can be applied to the fiber mass before extraction or strand or thread formation, or it can be sprayed onto the finished article. Both application methods are satisfactory as long as from 1 x 10~^ wt% or more of the antimicrobial material remains on the fabric. More than 0.1 - 1% by weight is generally too much and redundant.

Another use is alone or in solution or suspension or in conjunction with soaps or detergents for use in cleaning the skin, particularly in pre-surgical scrubbing preparations, or in preparations to combat the growth of Corynebacterium acnes. C. acnes is a species of bacteria implicated

in acne disorders, especially Acne vulgaris, where application of as little as 1-5 ppm is effective in combating such skin bacteria.

Higher concentrations can, if desired, be used without irritation or discomfort such as 2500 ppm and higher. When the cleaning preparation is diluted with water during use, the preparation can contain from 0.01% by weight and more of the polyamine according to the invention.

Moreover, the compounds described here can be used in enclosed water, such as swimming pools, ponds or industrially used water such as cooling or paper mill water to inhibit the growth of unwanted bacteria, fungi and/or algae.

When combating slime-forming microorganisms and algae in recirculating industrial water, especially cooling operations and special installations such as cooling towers, the polyamine compounds according to the invention are usually used alone, but can also be used in combination with other antimicrobial agents. The compounds are preferably used as salts to increase solubility. Concentrations in the recirculating water of as little as 1 x 10~^ wt% are sufficient to inhibit microbial growth. To ensure effectiveness, particularly against more resistant strains of microorganisms, and also when supplemental water is added to replace water loss through evaporation etc., concentrations of from 1 x 10-4 to 5 x 10% by weight are most satisfactory. The addition can be continuous or as periodic "shock treatment", i.e. addition for a 10 - 20 minute period every 4 - 8 hours.

An unusual, highly beneficial property of these compounds is their high adhesion to all kinds of surfaces, which provides protection against corrosion and acts as a storage depot for the continuous supply of water in contact with them. The same properties are also mainly responsible for the previously mentioned use as disinfectants for inanimate surfaces such as walls and ceilings, equipment, animal stalls, hospital facilities, kitchens and bathrooms, etc.

When working up the compounds according to the invention for the above applications, these compounds can be used in combination with other antimicrobial agents, surfactants, insecticides, defoamers, deodorants or as chelates of metals such as copper, calcium, magnesium and iron.

For agricultural applications, wettable powder preparations for use as a dispersion in water are a practical means of good distribution in the soil. Other methods of achieving the same results include the production of dust. All the polyamines can be mixed as fine powders with the commonly used powder diluents such as talc, clay, refined silicates, wood flour, sand, magnesium oxide, calcium carbonate, Fuller's earth, kaolin, diatomaceous earth, mica, pumice and the like. The powder can have the following composition:

The mixtures can be pulverized, e.g. to 1 - 10 µm average particle size, or they can be prepared by mixing the already finely powdered ingredients.

For use as agricultural disinfectants, the dusts can be applied to the seed and surrounding soil at the time of planting. The concentration of the sterilizing agent is adjusted to obtain an effective, non-phytotoxic dose in the soil. In general, the soil concentration of polyamine should be from 10 to 25 ppm (of active ingredient). For the most economical and effective application, the dusts can be applied in bands with 15 - 20 cm center distance on the rows just before sowing. The material can then be harrowed down to a depth of several centimeters. This method of treatment reduces material consumption and protects the root system of young plants against microbial attack. For the protection of a given plant, e.g. cabbage, the band distance of antimicrobial agent can vary from 20 cm for cabbage rot

to 30 - 38 cm for root ball prevention. Likewise, the depth to which the fungicide should be distributed can vary from 5 to 15 cm.

The wettable powders can be prepared by adding

0.1 - 5% wetting agent for the powder mixtures. Many dispersants are commercially available which are non-phytotoxic at the required concentrations. These can e.g. be alkali metal and amine salts of sulphated and sulphonated acids, alcohols and oils, or polyethoxylated alkylphenols, long fatty acid chain amine quaternary salts, partial fatty acid esters of polyhydric alcohols, etc. Some dispersants can be used in the production of emulsifiable concentrates of the polyamines in organic solvents. Many of these agents are available in solvent-soluble form. <p>The method of application to the soil is similar to that of powders. Spraying equipment is used to spread the suspensions or emulsions over the soil and with disc harrowing, the fungicidal agents can be distributed evenly to varying depths. Spray application is also effective for band-limiting the dosage.

Other agricultural uses for these preparations include eradication of bacterial diseases on plants by application to the involved surface areas. The compounds according to the invention show a high degree of bacterial inhibition and are particularly useful for this purpose. Some of the diseases which are of commercial importance in that they reduce yield and quality, and which are combated by the preparations according to the invention, are "fire blight" on apples and pears, bacterial spot on stone fruit, cherry leaf spot, hail-shot disease on walnut (caused by Pseudomonas juglandis ),

"common blight of bean", bacterial spot on tomato and pepper, and seedling potato rot. The effective concentration of polyamines required varies from 5 - 200 ppm. They can be applied as dust, powder dispersions in water or emulsions in water, or as aqueous dipping baths. Other plant diseases that can be combated by treatment with these preparations are of fungal origin, such as the many types of powdery mildew and leaf scab.

For seed treatment, amounts as low as 62 - 250 g/100 kg (550 - 600 ppm on seed) are effective against various fungi.

The compounds according to the invention can be used in the form of aqueous suspensions or emulsions when the base products are largely insoluble in water. Different powdered carriers can be used for this type of preparation to achieve an even distribution. Talc, Fuller's earth, calcium silicate, calcium carbonate, clays and the like are mixed with the agent together with wetting and dispersing agents and adhesives. For maximum chemical compatibility, those that are non-ionic in nature are preferred. Other nonionic or cationic surfactants are also satisfactory.

Other uses for the compounds according to the invention include inhibiting the formation of dental plaque, particularly when used as a mouthwash or in combination with toothpaste or tooth powder containing 50 - 1.000 ppm.

The following examples will further illustrate the invention.

Method A

Free-radical addition of acetic anhydride to [ 3-pinene

A mixture of 136 g (1.0 mol) p-pinene and 30 g (0.2 mol) t-butyl peroxide is added dropwise to 1,000 g (10 mol) of refluxing acetic anhydride over the course of 2.5 hours. The reaction mixture is then heated under reflux for a further 5 hours. The acetic anhydride is removed under vacuum, and the residue is hydrolysed by treatment with 40 g of sodium hydroxide in 250 ml of water and 150 ml of ethanol. The mixture is heated under reflux for 2 hours, then acidified with hydrochloric acid, extracted with ether and dried over sodium sulfate. The dried extracts are evaporated, whereby a residue is obtained which is distilled under vacuum, which gives 43.1 g (22%) of 3-(4-isopropylcyclohexenyl)-propionic acid with a boiling point of 135 - 137°C

(0.3mm).

Method B

3-(4-isopropylcyclohexyl)-propionic acid

The unsaturated acid from the previous method is dissolved in ethanol and hydrogenated with platinum oxide at room temperature and 2.8 kg/cm 2 hydrogen pressure. The platinum catalyst is filtered off, and the ethanol is removed under reduced pressure. The saturated product,

3-(4 isopropylcyclohexyl)-propionic acid is obtained as a colorless liquid in an amount of 42.3 g (97%).

Method C

Preparation of 1,5-di-(4-isopropylcyclohexyl)-3-pentanone

39.7 g (0.20 mol) of 3-(4-isopropylcyclohexyl)-propionic acid and 6.15 g (0.11 mol) of hydrogen-reduced iron are heated for 1.5 hours at

1°5°C under nitrogen. The temperature is then increased to 290°C, and this temperature is maintained for 3 hours. The cooled reaction mass is extracted well with ether, filtered through "Celite", and the ether extracts are evaporated under vacuum. The residue is stripped under vacuum to give the product, 17.39 (51%).

In a similar way, the following ketones are obtained: 1,9-dicyclohexyl-5-nonanone; 1,5-dicyclohexyl-3-pentanone; 1,3-dicyclohexylacetone; 1,7-dicyclohexyl-4-heptanone; 1,3-di-(3-methylcyclohexyl)-acetone; 1,7-di-(4-ethylcyclohexyl)-4-heptanone; 1,5-di-(2-isopropylcyclohexyl)-3-pentanone; 1,9-di-(2-ethylcyclohexyl)-5-nonanone; 1,5-di-(4-t-butylcyclohexyl)-3-pentanone; 1,5-di-(2,4,6-trimethylcyclohexyl)-3-pentanone; 1,5-di-(3,5-diethylcyclohexyl)-3-pentanone; 1,7-di-(2,6-dimethyl-4-t-butylcyclohexyl)-4-heptanone; 1,7-di-(2,3,4,5,6-pent amethylcyclohexyl)-4-heptanone; when unsaturated acids are subjected to the above treatment, the following representative ketones are obtained: 1.7-dicyclohex-3-enyl-4-heptanone;

2,8-di-(4-methylcyclohex-3-enyl)-5-nonanone and 1,5-di-[4-isopropylcyclohex-1-enyl]-3-pentanone.

Method D

Preparation of 4-cyclohexy1-1-(4-isopropylcyclohexyl)-butanone-2

A Grignard reagent was prepared from 21 g (0.11 mol) of 2-cyclohexyl ethyl bromide and 2.4 g (0.1 g atom) of magnesium. The magnesium was covered with 25 ml of anhydrous ether, an iodine crystal was added and, in a nitrogen atmosphere, the halide dissolved in 50 ml of anhydrous ether was added over the course of 1 - 2 hours as the temperature, when the initial reaction had occurred, was raised to the reflux temperature. After the addition is finished, the reflux is continued for 1/2 hour.

In a nitrogen atmosphere, the Grignard solution is clarified by passing it through a glass wool filter plug and slowly added to a stirred solution of 14.9 g (0.09 mol) 4-isopropylcyclohexyl acetonitrile in 200 ml of anhydrous diethyl ether. A gentle reflux is maintained during the addition which takes 1/2 to 1 hour. After completion of the addition and a further 15 minutes of reflux, the reaction mixture is cooled and poured onto a

mixture of 50 ml of concentrated hydrochloric acid and 200 g of ice while mixing well. During heating, the ether is removed by distillation, and the residue is heated at 70 - 100°C for 1 hour. The product is extracted with 2 x 250 ml of ether, the ether solution is dried over anhydrous magnesium sulfate, and the solvent is removed. Any remaining reactants, i.e. halide and nitrile, are removed from the ketone by fractional distillation under reduced pressure together with by-products.

In a similar manner, the following ketones are prepared: 1-(2-methylcyclohexyl)-4-cyclohexylpentan-2-one;

1-(4-t-butylcyclohexyl)-5-(4-isopropylcyclohexyl)-pentan-3-one;

2-(3-methylcyclohexenyl)-8-(2-isopropylcyclohexyl)-octan-4-one;

1-(2,6-dimethyl-4-t-butylcyclohexyl)-5-(3,5-diethylcyclohexyl)-pentan-3-one.

Method E

Preparation of N-(3-aminopropyl)-1,4-cyclohexane-bis-(methylamine)

26.5 g (0.5 mol) of acrylonitrile is added dropwise over 45 minutes to 2.84 g (2.0 mol) of 1,4-cyclohexane-bis-(methylamine) while stirring and cooling in an ice bath. After the addition is complete, the reaction mixture is stirred for a further 1 hour at 5°C, gradually heated to 45°C and held for 2 hours at this temperature, followed by 1 hour at 90°C. Any unreacted acrylonitrile and excess of non-cyanoethylated bis-(methylamine) starting material which was removed at an internal temperature of 110°C and 1 mm are driven off from the reaction mixture. The residue is then dissolved in 1.5 1 of ethyl alcohol (saturated with ammonia gas) mixed with 50 ml of sponge nickel catalyst and hydrogenated at 10.5 kg/cm 2 . After removal of the catalyst by filtration, the solvent is driven off, and the ammonia and triamine product are purified by fractionation under reduced pressure.

A higher homologue, N-(3-aminopropyl)-1,4-cyclohexane-bis-(2-ethylamine), is synthesized using the above method with 1,4-bis-(2-aminoethyl)-cyclohexane prepared according to

P.P. Garcia and J.H. Wood, J. Org. Chem., 26, 4167 (1061). Excess starting amine in this example can be separated from the product at a boiling point of 122 - 126°C/l mm.

Method F

Preparation of N-(3-aminopropyl)-N'-(2-hydroxyethyl)-1,4-cyclohexane-bis-(methylamine)

10.6 g (0.2 mol) acrylonitrile is added dropwise over 15 minutes to 37.2 g (0.4 mol) N-(2-hydroxyethyl)-1,4-cyclohexane-bis-(methylamine) while stirring and ice bath cooling. After the addition is complete, the reaction mixture is stirred for a further 2 hours at 5°C, allowed to gradually heat up over the course of 1 hour, heated for 2 hours at 45°C and finally for one hour at 90°C. It is then fractionated under reduced pressure up to an internal temperature of 170°C. The residue is dissolved in 200 ml of ethyl alcohol, cooled in an ice bath and saturated with ammonia gas at 0°C. About. 5 ml of sponge nickel catalyst (W.R. Grace Co., Davison Chem. Division) is added and the mixture is shaken over hydrogen at 10.5 kg/cm until no more hydrogen is taken up. The catalyst is removed by suction filtration under nitrogen, and the solvent is driven off and the residue is subjected to fractional distillation under reduced pressure. The triamine product can be easily distinguished from the cyanoethylated diamine by its lower R^. on silica gel using a solution of one volume of concentrated aqueous ammonium hydroxide in 4 volumes of methyl alcohol. The synthesis is adapted according to the method of M. Israel et al., J. Med. Chem., 7, 710 (1964) for the preparation of polymethylene polyamines.

Method G

Preparation of N-(2-hydroxyethyl)-1,4-cyclohexane-bis-(methylamine)

A solution of 14.2 g (0.1 mol) of 1,4-cyclohexane-bis-(methylamine) in 150 ml of anhydrous methyl alcohol and under a nitrogen atmosphere is heated to 45-50°C. During 20 minutes, a total of 1.1 g (0.025 mol) of ethylene oxide in gaseous form is introduced under good stirring and under the surface of the liquid. The reaction temperature is kept at 45 - 50°C for a further 1/2 hour after the addition of ethylene oxide has ended. The methyl alcohol is removed by distillation at atmospheric pressure, and excess 1,4-cyclohexane-bis-(methylamine) is easily separated from the product by fractionation under reduced pressure. Only the monoethoxylated compound remains and can be used as such or further purified by distillation under reduced pressure.

Method H

Preparation of N-(3-amino-2-hydroxypropyl)-1,4-cyclohexane-bis-(methylamine) _____

14.2 g (0.1 mol) of 1,4-cyclohexane-bis-(methylamine) are dissolved in 50 ml of anhydrous methyl alcohol, and the solution is cooled to +5°C in an ice bath. 9.3 g (0.1 mol) of epichlorohydrin are added over 2 minutes, and the temperature is maintained at +5°C for 2 hours. The reaction is allowed to proceed at 10 - 15°C until thin layer chromatography of a sample (silica gel plate developed using a solution of 1 volume concentrated aqueous ammonium hydroxide 1 4 volumes methyl alcohol) showed almost complete transfer of the starting diamine to the propylene chlorohydrin. The solution is then added to 100 ml of dry methyl alcohol which has previously been saturated at 0°C with dry ammonia gas by continuous dropwise addition at +5°C with good stirring and external cooling. After stirring for 2 hours at +5°C, it is allowed to warm to 20°C and mixed overnight. The reaction is completed by heating at 45 - 55°C for 6 hours. The solvent and ammonia were removed by stripping and the product purified using fractional distillation under reduced pressure.

N-(2,3-dihydroxypropyl)-1,4-cyclohexane-bis-(methylamine) is prepared by alkaline hydrolysis of the above propylene chlorohydrin derivative.

The propylene chlorohydrin derivative is dissolved in an IM sodium hydroxide solution containing 50% by weight of methyl alcohol and 50% by weight of water in a ratio of 5 g of chlorohydrin to 25 ml of sodium hydroxide solution. After stirring for 24 hours at 20°C, the methyl alcohol is removed by distillation, and the oil that separates is extracted with 100 ml of diethyl ether. The extract is washed with approx. 10 ml of cold water, the ether layer is dried over anhydrous sodium sulphate and then filtered. Removal of the ether by distillation gives the product of good purity as an oil.

Method I

Preparation of N,N'-bis-(3-aminopropyl)-1,4-bis-(2-aminoethyl)-cyclohexane

10.6 g (0.2 mol) acrylonitrile is added dropwise during

15 minutes to 17.0 g (0.1 mol) of 1,4-bis-(2-aminoethyl)-cyclohexane cooled in an ice bath and with good stirring. The resulting solution is kept at 5-10°C with stirring for 1 hour, allowed to warm to 25°C over 2 hours and finally heated at 90-95°C for 4 hours. The reaction mixture is then freed of any unreacted material and monocyanoethylated product by gradual heating to an internal temperature of 130°C at a pressure of

0.5 - 1 mm. The residue is dissolved in 200 ml of ethyl alcohol which has previously been saturated with dry ammonia gas at 0°C, mixed with approx. 5 ml of a sponge-nickel catalyst suspension and reduced by shaking under 14 kg/cm hydrogen thrust. The catalyst is removed by suction filtration, the filtrate is stripped of solvent, and the residue is purified by fractional distillation under reduced pressure.

Method J

Preparation of N-(2-aminoethyl)-1,4-bis-(2-aminoethyl)-cyclohexane

°8 g (0.4 mol) 1,4-bis-(2-aminoethyl)-cyclohexane and 4.3 9 (0.1 mol) ethyleneimine with 0.4 9 ammonium chloride are mixed in a glass-lined pressure reactor and filled with nitrogen to 7 kg/cm 2. The mixture is shaken and heated at 85 - 95°C for 48 hours. After cooling, it is quickly distilled free of the salt and then fractionated under high vacuum. The starting diamine is easily separated from the triamine product by thin-layer chromatography on silica gel using a mixture of 1 volume of concentrated aqueous ammonium hydroxide with 4 volumes of methyl alcohol, the diamine having a much higher R^..

Method K

N,N-bis-(3-hydroxypropyl)-1,4- cyclohexane-bis-(methylamine)

Preparation of 1-cyano-4~[di-(3-hydroxypropyl)-aminomethyl]-cyclohexane and catalytic reduction

a. 20.2 g (0.1 mol) 1-bromomethyl-4-cyanocyclohexane and

53.2 g (0.4 mol) of di-(3-hydroxypropyl)-amine in 400 ml of anhydrous isopropyl alcohol are heated in an autoclave at 105 - 115°C for 8 hours with continuous stirring. The reaction mixture is stripped of solvent under reduced pressure, and the residue is diluted with 500 ml of ice water. A cold solution of 5 g of sodium hydroxide in 100 ml of water is added, and the mixture is extracted with 2 x 150 ml of methylene chloride. The organic phase is then washed with 50 ml of ice water, dried overnight with anhydrous sodium sulfate, filtered and freed from solvent by distillation under reduced pressure.

b. The residual oil from a.) is taken up in 200 ral of anhydrous ethyl alcohol which has previously been saturated at 0°C with dry ammonia gas, mixed with 5 ml of sponge-nickel catalyst suspension and hydrogenated at 25°C under 7 kg/cm2 hydrogen pressure in a stirred autoclave . The end of the reaction is easily determined by the disappearance of the C-N IR absorption band and measurement of the hydrogen uptake. The catalyst is removed by suction, the solvent by mild heating under reduced pressure, and the product is obtained pure by fractional distillation under reduced pressure.

Method L

N,N-di-(2,3-dihydroxypropyl)trimethylenediamine

16.5 g (0.1 mol) bis-(2,3-dihydroxypropyl)amine and 6.4 g (0.12 mol) acrylonitrile were mixed in an ice bath and then warmed to room temperature. After standing for 2 hours, the mixture was heated at 45 - 55°C for 3 hours. The excess of acrylonitrile was removed by careful heating under reduced pressure. The residue was taken up in ethyl alcohol, mixed with sponge nickel catalyst and hydrogenated under 14 kg/cm hydrogen pressure with good stirring. After filtering off the catalyst, the solvent and excess acrylonitrile were removed by stripping under reduced pressure, whereby the product was obtained as an oil.

Method M

N,N,N'-tri-(2,3-dihydroxypropyl)-trimethylenediamine

11.1 g (0.05 mol) of N,N-di-(2,3-dihydroxypropyl)-trimethylenediamine was dissolved in 125 ml of methanol and heated under reflux with stirring. 3.7 g (0.05 mol) of glycidol was added dropwise over 1.5 hours, and the solution was stirred for a further 1 hour at 60-80°C. The methyl alcohol and other volatile substances ■ were removed by stripping under reduced pressure, whereby the product was made suitable for use in the next steps.

Method N

5, 9, 9- tri-(2, 3- dihydroxypropyl)- 1, 5, Q- triazanonan

A sample of 5.9 g (0.02 mol) of the residual oil from method M was mixed with 2.75 g (0.05 mol) of acrylonitrile at room temperature and then heated at 50-6o°C for 10-15 hours. Excess acrylonitrile was removed by stripping under reduced pressure, and the residual oil was taken up in 50 ml of ethanol, mixed with 2 g of sponge nickel catalyst and shaken in a hydrogen atmosphere of o 14 kg/cm 2 for 6 hours. The mixture was filtered free of catalyst, and the solvent was removed by distillation. The product could be brought to analytical purity by chromatography on a silica gel column and was an oil.

Example 1

First Ilation of 1-[1,5-di-(4-isopropylcyclohexyl)-3-penty1]-1,5,9-triazanonane

6.70 g (0.02 mol) of 1,5-di-(4-isopropylcyclohexyl)-3-pentanone and 13.1 g (0.10 mol) of 3,3'-imino-bis-propylamine in 150 ml of toluene are heated under reflux overnight with a Dean-Stark water separator. The cooled solution is evaporated under reduced pressure. The residue is dissolved in ethanol and hydrogenated with platinum oxide at room temperature and 2.8 kg/cm hydrogen pressure. The platinum catalyst is filtered off and the ethanol is removed under vacuum. The remaining oil is dissolved in ether, and the ether solution is washed several times with water to remove excess 3,3*-imino-bis-propylamine.The ether extracts are dried over anhydrous sodium sulfate and evaporated under vacuum to give the polyamine as a colorless oil.

The oil is dissolved in ether, and hydrogen chloride gas is bubbled into the solution until no further refining occurs. The ether is evaporated under reduced pressure and gives the product as a solid which is dissolved in hot isopropyl alcohol. The solids are collected by filtration and dried under vacuum at 70°C, whereby a colorless product is obtained, 1-[1,5-di-(4-isopropylcyclohexyl)-3-pentyl]-1,5,9-triazanone trihydrochloride .

Analogously, from the ketones and amines indicated below, the following compounds according to the invention are prepared:

Example 2

Preparation of 1-[1,5-di-(4-isopropylcyclohexen-1-yl)-3-pentyl]-. 1 , 5 , 9-t riazanonane

6.60 g (0.02 mol) of 1,5-(4-isopropylcyclohexen-yl)-3-pentanone and 13.1 g (0.10 mol) of 3,3'-imino-bis-propylamine in 150 ml of toluene heated under reflux overnight with a Dean-Stark water separator. The toluene is then removed under vacuum. The remaining oil is dissolved in 25 ml of isopropanol and added dropwise to 1.90 g (0.05 mol, excess) of sodium borohydride suspended in 50 ml of isopropanol. After the addition is complete, the reaction mixture is heated under reflux for 1 hour. The isopropanol is evaporated under reduced pressure, the residue is treated with water, and the aqueous mixture is extracted well with ether. The combined ether extract is washed back

with water, a saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated under vacuum to give the polyamine product as a clear oil in an amount of 7.4 <3 (90%).

The oil is dissolved in ether, and the solution is cooled in an ice-water bath. Hydrogen chloride gas is bubbled into the solution until no further precipitate forms. The solid is collected by filtration, washed with a small amount of ether and dried under vacuum, whereby the polyamine trihydrochloride is obtained as a colorless product (96%), m.p. 256-257°C.

In an analogous manner using the ketones and amines indicated below, the following compounds according to the invention are prepared.

Example 3

l-[ 1, 7- di-( 4- methylcyclohexyl)- 4- heptyl]- 1, 4, 8- triazaoctane

A mixture of 0.03 mol of 1,7-di-(4-methylcyclohex-3-enyl)-4-heptanone and 12.0 g (0.20 mol) of 1,2-diaminoethane in 250 ml of ethanol is heated under reflux over the night. The cooled reaction mixture is hydrogenated with platinum oxide at room temperature and 2.8 kg/cm hydrogen pressure. The platinum catalyst is filtered off, and the ethanol is removed under reduced pressure. The residual oil is dissolved in ether, and the ether solution is washed several times with water to remove excess diaminoethane. The ether extracts are dried over anhydrous sodium sulfate and evaporated under vacuum, which gives 11.2 g (100%) of a colorless oil.

The oil is dissolved in 20 ml of t-butanol and cooled to 0-5°C in an ice-water bath. 1.75 g (2.2 ml, 0.033 mol) of acrylonitrile is added dropwise over 5 minutes. The reaction mixture is allowed to warm to room temperature and then heated at 60°C overnight. The t-butanol is removed under reduced pressure. The remaining oil is dissolved in 150 ml of glacial acetic acid and hydrogenated with platinum oxide at room temperature and 2.8 kg/cm 2 hydrogen pressure. The platinum catalyst is filtered off, and the acetic acid is removed under vacuum. The residue is dissolved in ether and made alkaline with 10% sodium hydroxide. The ether solution is washed with water, dried over anhydrous sodium sulfate and evaporated under reduced pressure to obtain the product.

Example 4

Preparation of 1-[1,5-di-(4-isopropylcyclohexyl)-3-pentyl]-5-(2,3-dihydroxypropyl)-1,5,9-triazanonane

6.7 g (0.02 mol) 1,5-di-(4-isopropylcyclohexyl)-3-pentanone and 20.5 g (0.10 mol) 3,3'-(2,3-dihydroxypropylimino)-bis -propy1-amine (obtained by catalytic hydrogenation of dicyanoethylated glycerylamine, in 150 ml of toluene is heated under reflux overnight with a Dean-Stark water separator. The cooled solution is evaporated under reduced pressure. The residue is dissolved in ethanol and hydrogenated with platinum oxide at room temperature and 2 .8 kg/cm hydrogen pressure. The platinum catalyst is filtered off, and the ethanol is removed under vacuum. The remaining oil is dissolved in ether, and the ether solution is washed several times with water to remove excess 3,3'-(2,3-dihydroxypropylimino)-bis -propylamine The ether extracts are dried over anhydrous sodium sulfate and evaporated, whereby the polyamine product is obtained as an oil.

In a similar manner and using analogous amounts, but using N,N-di-(2,3-dihydroxypropyl)-trimethylenediamine and 5,9,9-tri-(2,3-dihydroxypropyl)-1,5, 9-triazanone instead of 3,3'-(2,3-dihydroxypropylimino)-bis-propylamine is produced respectively. N-[1,5-di-(4-isopropylcyclohexyl)-3-pentyl]-N*-di-(2,3-dihydroxy-propyl)-trimethylenediamine and 1-[1,5-di-(4-isopropylcyclohexyl) )-3-pentyl]-5-(2,3-dihydroxypropyl)-9-di-(2,3-dihydroxypropyl)-1,5,9-triazanonane.

Example 5

Preparation of 1-[1,5-di-(4-isopropylcyclohexyl)-3-pentyl]-5-(2,3-dihydroxypropyl)-9-(1,3-dihydroxy-2-propyl)-1,5, 9- triazanone

5.2 g (0.01 mol) 1-[1,5-di-(4-isopropylcyclohexyl)-3-pentyl]-5-(2,3-dihydroxypropyl)-1,5,9-triazanonane and 9 g (0.1 mol) of 1,3-dihydroxyacetone in 100 ml of chloroform was heated under reflux with a water separator connected until 1.8 ml of water was collected (8-12 hours). The chloroform and excess 1,3-dihydroxyacetone were removed by distillation under reduced pressure. The remaining oil was taken up in 75 ml of ethanol, mixed with 1 g of platinum oxide and hydrogenated at 2.8 kg/cm hydrogen pressure while shaking at room temperature. The catalyst was removed by filtration and the ethyl alcohol by distillation to give an oil. The product could be purified by column chromatography using silica gel and

development with methyl alcohol containing ammonium hydroxide.

In an analogous way, but starting from 1,7-di-(2,3-dimethyl-cyclohexyl-4-heptyl)-ethylenediamine instead of 1-[1,5-di-(4-iso-propylcyclohexyl)-3 -pentyl]-1,5,9-triazanonane, N-[1,7-di-(2,3-dimethylcyclohexyl)-4-heptyl-N<*->(1,3-dihydroxy-2-propyl) is obtained -ethylenediamine.

Example 6

1-[1,7-di-(4-methylcyclohexyl)-4-heptyl]-4,8,8-tri -(2,3-dihydroxy-propyl)- 1, 4, 8- triazaoctane

4.1 g (0.01 mol) of 1-[1,7-di-(4-methylcyclohexyl)-4-heptyl]-1,4,8-triazaoctane was dissolved in 50 ml of methanol and heated under reflux and stirring. 15 g (0.2 mol) glycidol was added dropwise over 1.5 - 2 hours. After completion of the addition, the reaction mixture was stirred for a further 2 hours at 90 - 100°C. The methyl alcohol was removed by stripping under reduced pressure and excess glycidol by distillation at 1 mm pressure. The residue could be further purified by transfer to the trihydrochloride salt in ethyl alcohol with dry hydrogen chloride and fractional crystallization. The free base can then be released from its salt by resin ion exchange or neutralization with aqueous sodium hydroxide.

Analogously using the following dicyclohexyl polyamines, the following products are obtained:

Also, each of the respective ketones IV given in methods C and D, when reacted with each of the individual amines given on page 11, last paragraph, first according to the procedure given in Example 1, and then according to Example 2 , the entire range of compounds described according to the present invention as shown by formula I.

Claims (4)

1. Amines for use as antimicrobial and/or algae inhibiting agents, characterized in that they have the formula: where A, which are the same or different, is cyclohexyl or cyclohexenyl of the formula: where each R is either hydrogen or lower alkyl, each R 1 , which is the same or different, is alkylene of 1-4 carbon atoms; and Z is -(CH 2 ) 3 NH(CH 2 ) 3 NH 2 ; -(CH 2 ) 2 NH(CH 2 ) 2 NH(CH 2 ) 2 NH 2 ; - (CH 2 ) 3 NCH 3 (CH 2 ) 3 NH 2 ; -CH2CHOHCH2NH2; -(CH^NE^; -(CH2)2NH2 or -CH2"CHOH-NH2 and acid addition salts thereof. 2. Connection according to claim -1-, characterized in that it is 1-[1,5-di-(4-iso-propylcyclohexyl)-3-pentyl]-1,5,9-triazanonane. 3. Connection according to claim 1, characterized in that it is 1-[1,5-di-(4-iso-propylcyclohex-1-enyl)-3-pentyl]-1,5,9-triazanonane. 4. Compound according to claim 1, characterized in that it is 1-[1,9-dicyclohexyl-5-nonyl]-1,5,9-triazanonane trihydrochloride.