DE1274281B - Germicide - Google Patents

Description

FEDERAL REPUBLIC OF GERMANY GERMAN W7TW> PATENT OFFICE Int. Cl .:

A611

EDITORIAL

A61k
C07c German class: 30-3
30 h-13/10
12 q-32/21 Number: 1274281 File number: P 12 74 281.3-41 (U 13316) Registration date: November 30, 1966 Display day: !. August 1968

The present invention relates to germicides which use a quaternary ammonium compound as an active ingredient contain.

Quaternary ammonium compounds are widely used as germicides because of their recognized microwave activity. They possess both bacteriostatic as well as bactericidal properties and also considerable surface activity. Because of these properties Quaternary ammonium compounds have wide application for the sterilization of surgical and dental instruments, cutlery, food processing machines and other equipment found for the food industry. They are also used for the treatment of small wounds, the post-operative sterilization of human tissue, and as an antiseptic additive to mouthwashes and other products for human hygiene.

The production of a certain group of germicidal quaternary ammonium compounds, namely those characterized by the presence of a long-chain alkoxyalkylene group in the molecule is in Swiss patents 301 409 and 303 510 to 303 514 described.

However, the majority of these compounds are bitter and have an unpleasant taste, which is disadvantageous. when these compounds are to be added to agents that are used in the mouth will. The compounds used according to the invention do not have this disadvantage.

The quaternary ammonium compounds of the present invention have the general formula

R ¹

Germicide

R ³ CHOH CH ₇ -NR ²

CH,

in which R ^{1 is} a C1 to Gj alkyl group or a C2 to Gr hydroxyalkyl group. R- is a C1 to G; -alkyl group or a G> - to Gi-hydroxyalkyl group. R ^:! a Gr to C 1-4 alkyl group. X is hydrogen or halo and Y is a halo ion. means.

Applicant:

Unilever N.V. Rotterdam (Netherlands)

Representative:

Dr.-Ing. A. van der Werth and Dr. F. Lederer,

Patent attorneys,

8000 Munich 80, Lucile-Grahn-Str. 22nd

Named as inventor:

Thaddeus John Kaniecki, Brooklyn, N.Y .;

Arno Cahn, Pearl River, N.Y. (V. St. A.)

Claimed priority:
V. St. v. America December 2, 1965
(511247)

These compounds differ from the previously known quaternary ammonium compounds in that they have one in the longest alkyl chain which is on the nitrogen atom Have 2-hydroxy substituents.

Examples of such quaternary ammonium compounds are N-methyl-N- (2-hydroxyethyl) -N- (2-hydroxytetradecyl ) -N-benzyl-ammonium chloride, N-methyl - N - (2 - hydroxyaryl) - N - (2 - hydroxydecyl) -N - benzyl - ammonium chloride, N - methyl - N - (2 - hydroxyethyl) - (2-hydroxyhexadecyl) -N-benzyl-ammo- nium chloride.

Preferred quaternary ammonium compounds are N-methyl-N- (2-hydroxyethyl) -N- (2-hydroxydodecyl) -N-benzylammonium chloride and N-methyl - N - (2 - hydroxyethyl) - N - (2 - hydroxydodecyl) -N- ( p-brom obenzyl) -amm oni umbrom id.

The new 2-hydroxyalkylated quaternary ammonium compounds can be prepared by condensation of a suitable 1,2-epoxyalkane with an equimolar amount of a suitable secondary amine to form a tertiary Amine, which is characterized by the presence of a 2-hydroxy group on the longest alkyl chain that is is located on the nitrogen atom. The associated quaternary ammonium

809 588/446

Binding is then established by condensation of the tertiary amine so produced with an equimolar Amount of a suitable benzyl halide.

Suitable 1,2-epoxyalkanes used for the production the tertiary amines can be used, which in turn then give the new 2-hydroxylated quaternary ammonium compounds can be implemented are 1,2-epoxy octane, 1,2-epoxynonane, 1,2 - epoxydecane, 1,2 - epoxyundecane, 1,2-epoxydodecane, 1,2-epoxytridecane, 1,2-epoxytetradecane, 1,2-epoxypentadecane, 1,2-epoxyhexadecane and the like, as well as mixtures of these compounds, z. B. Mixtures of epoxyalkanes, both an even and an odd Have number of carbon atoms in the alkyl chain.

Suitable secondary amines are dimethylamine, diethylamine, methylethylamine, diethanolamine, dipropanolamine, Methyl monoethanolamine and the like.

Organic benzyl halides suitable for condensation with the tertiary amines mentioned above are suitable are, for. B. benzyl chloride, benzyl bromide, benzyl iodide, p-bromobenzyl bromide, p-chlorobenzyl chloride.

The compounds described here, for the sake of simplicity, referred to as antiseptic agents Can be beneficial in bacteriologically effective amounts in many hygienic areas Means such as B. Mouthwashes and cleaning solutions for sterilization of surgical and dental instruments or objects and containers used in food processing will. "Bacteriologically effective amount" is understood to mean the amount that a certain compound is needed to kill the microorganisms or their growth to inhibit. Various factors, such as the possible contact time, the type of microorganisms and the conditions under which the antiseptic is used will determine the amount that will be used as bacteriologically effective amount can be designated.

The invention is illustrated by the following examples.

The production of certain N-methyl-N- (2-hydroxyethyl) -N- (2-hydroxyalkyr) -am and N, N-bis (2-hydroxyethyl) -N- (2-hydroxyalkyl) amines are described first.

Table I. Alkyl group (R ³ ) yield
° / o C10H21
C12H25
C14H29 98.8
95.8
99

Condensation of 1,2-epoxyalkanes with diethanolamine

Table II below shows results obtained with a number of N, N-bis (2-hydroxyethyl) -N- (2-hydroxyalkyl) amines obtained by condensation of 1,2-epoxyalkanes with 12 to 18 Carbon atoms were made with diethanolamine. In each case, 0.3 mole of diethanolamine and 0.3 mole of a 1,2-epoxyalkane were dissolved in 50 to 100 ml of ethyl alcohol and the resulting mixture was heated on a steam bath with occasional stirring until a homogeneous solution was obtained. Heating was then continued until the ethyl alcohol was distilled off, the last traces were removed at 7O ⁰ C under a pressure of 30 mm Hg. In each case the product was cooled to room temperature and the melting point of a sample recrystallized from ethyl acetate was determined. The alkyl groups mentioned in column 1 of Table II correspond to group R ^{3 of} the above general formula.

Table II

Alkyl group (R ³ ) yield
% Melting point
⁰ C C10H21
Q2H20
C14H29
C16H33 99
99
96.5
80 *) 36 to 38
49.5 to 50
51 to 52
59 to 60

Condensation of 1,2-epoxyalkanes with methyl monoethanolamine

N-methyl-N- (2-hydroxyethyl) -N- (2-hydroxyalkyI) -amines were prepared by heating a Reaction mixture consisting of 0.5 mol of a 1,2-epoxyalkane and 0.5 mol of methyl monoethanolamine, dissolved in about 75 ml of ethyl alcohol, on a steam bath until homogeneous Solution. The temperature was then increased and the ethyl alcohol was distilled off. Rest Traces of ethyl alcohol were removed under reduced pressure.

Table I shows the yields of various N-methyl-N- (2-hydroxyethyl) -N- (2-hydroxyalkyl) amines which were obtained in this way. The alkyl groups which are mentioned in the first column of Table I are the groups R ^{3 of} the above general formula.

*) On recrystallized material.

example 1

N-Methyl-N- (2-hydroxyethyl) -N- (2-hydroxyalkyl) -N-benzylammonium chlorides were prepared by adding equimolar amounts (0.05 mol) of a tertiary amine, which had been prepared as described above, and of benzyl chloride in 40 ml of ethanol were refluxed. The reaction mixture was kept for 24 hours at 50 ⁰ C under reflux and then cooled to room temperature and kept at room temperature for 10 days. After this time the ethanol was removed and the reaction product was stored over concentrated sulfuric acid at a pressure of 200 mm Hg until the product crystallized.

Table III contains information on the substances produced in this way. The alkyl groups mentioned in column 1 of the table correspond to the group R ^{3 of} the above general formula

Table III

Alkyl group (R ³ ) yield
% Cationic
% C10H21
C ₁₂ H ₂₅
C14H2 » 97
94.7
99.5 91.4
94.7
92.4

Example 2

N- (hydroxydodecyl) -N-methylethanolamine (0.05 mol) and p -Bromobenzylbromid (0.05 mol) dissolved in 40 ml of ethanol and kept together at 50 ⁰ C for 24 hours under reflux. The resulting reaction mixture was cooled and held at room temperature for 10 days, after which the ethanol was removed and the product crystallized over concentrated sulfuric acid at a pressure of 200 mm Hg.

Example 3

N ₅ N - bis (2 - hydroxyethyl) - N - (2 - hydroxyalkyl) -N-benzyl-ammonium chlorides were prepared by adding equimolar amounts (0.1 mol) of N- (2-hydroxyalkyl) -diethanolamine, which as described above, and benzyl chloride in 75 ml of n-propyl alcohol was refluxed for 6 hours. The n-propyl alcohol was then evaporated. The last traces of n-propyl alcohol were removed by heating the reaction product to 54 ° C. at a pressure of 30 mm Hg. The product thus obtained was then dried over concentrated sulfuric acid at a pressure of 0.5 mm Hg.

Table IV gives the values of the fabrics produced in this way.

Table IV

Alkyl group (R ³ ) yield
% Cationic
% C10H21
C12H25 81
69.7 54.5
97

N-methyl-N- (2-hydroxyethyl) -N- (2-hydroxydodecyl) -N-benzylammonium chloride (I) the present invention was noted for its germicidal activity against a wide range of microorganisms tested. Parallel tests were carried out with N-benzyl-N, N-bis (2-hydroxyethyl) dodecyloxyethyl ammonium chloride (II) and N-benzyl-N, N-bis (2-hydroxyethyl) decyloxyethyl ammonium chloride (III). The preparation of the compound II is described in the aforementioned Swiss patent 303 513, and the connection III is a homologous compound that only differs in terms of the chain length of the alkoxyalkylene group differs from compound II.

The minimum effective concentration of each of the quaternary ammonium compounds listed in Table V was determined using inclined plates using agar plates showing a gradual proportional increase in the concentration of the antiseptic compound along the axis of inclination. Such plates were doused with two layers of nutrient medium, the lower layer consisting of pure nutrient medium which was allowed to harden while the plate was tilted so that a wedge-shaped layer was formed. A second nutrient layer containing the antiseptic was then poured onto the plate while the plate lay flat. The downward diffusion of the antiseptic agent forms a uniform linear concentration gradient during the incubation. The plates were inoculated by brushing a suspension of a bacterial culture over the surface of the nutrient medium. The plates were ^{incubated at 37 °} C. overnight and the extent of the bacterial growth was measured. The minimum effective concentration against the respective microorganism was then calculated.

Table V

Js Se Ba Effective minimum concentration in ppm against- Sc Ps Po Approx 1.4 1.55 2.7 Mc Ec 200 265 15th Sat 0.67 1.0 1.55 Sf 2.6 44 255 410 2.2 4.45 3.0 2.2 1.2 2.0 5.0 2.5 275 59 194 1.8 2.6 2.0 3.15 5.0 43 11.7 5.4

Compound I.
Compound II.
Compound III

In the table means
Sa = Staphylococcus aureus
Js = Staphylococcus epidermidis
Se = Serratia
Ba = Bacillus aerogenes
Sf = Streptococcus faecelis

Mc = micrococcus
Ec = Escherichia coli
Sc = Salmonella choleraesius
Ps = Pseudomonas
Po = pityrosporium ovale

Ca = Candida albicans

An = Aspergillus niger

Table V are therefore comparative tests between a compound I according to the present invention Invention and compounds II and III can be found in the aforementioned Swiss patents, and it can be seen that e.g. B. the germicidal activity of compound I against E. Coli (Ec) is significantly better than the corresponding activity of the known compound II, while at the same time the germicidal activity of the compound I compared to S. aureus (Sa) is significantly better than that of compound IH.

A group of quaternary ammonium compounds that have a 2-hydroxy substituent in the longest alkyl chain was tested for their germicidal activity in a simulated mouthwash

tested, which from a 23% alcohol-in-water solution duration. The results of the experiments are summarized in Table VI below. Parallel tests with a known antiseptic agent were used as a basis for comparison.

The bacteriostatic activity was determined by an agar dilution cup test (ADC), including a standard Petri dish was filled with 40 ml of thioglycollate agar, which was l% with a culture was vaccinated by S. aureus. After hardening, a cylindrical plug was cut out of the agar, so that the agar formed a well into which a 10% agar dilution of the mouthwash was pipetted which contained the quaternary ammonium compound to be evaluated, so that the removed plug was washed through an agar-mouthwash mixture was replaced. After the agar-mouthwash mixture had hardened, the plate was incubated overnight at 37 ° C. Zones of inhibition, which developed during the incubation were measured and after subtracting the Applied diameter of the plug.

The substantivity of these selected compounds in a mouthwash was determined by a Skin-Disc-Substantivity-Test (SDS), in which a 10 ml germinal layer of S. aureus on a base of 40 ml of sterile thioglycollate agar was poured into a Petri dish. Calf skin slices were then hydrated in distilled water, agitated in the mouthwash solution, and thoroughly immersed in water rinsed for 15 minutes, excess water was removed from the slices, which were then placed on the surface of the prepared germination agar plates became. The plates were incubated overnight at a temperature of 37 ° C. The zones of inhibition, that formed during the incubation were measured and after subtracting the diameter of the Disc registered. If no zones of inhibition were evident, the disks were removed, and the inhibition on the contact area was evaluated.

The bactericidal activity was determined by a bactericidal contact time test (BCT) in which the whole saliva was collected from a group of donors and 1 ml of the collected saliva was mixed with 1 ml of the mouthwash in a glass vial at room temperature. Equal amounts (0.001 ml) of the mixture were withdrawn at certain time intervals and mixed with 5 ml of a sterile, liquid microinoculum agar at 45.degree. After hardening, the resulting mixture was ^{incubated at 37 °} C. to develop colonies. Counts were made in the same manner as the standard plate counts. The original inoculation density was calculated from quadruple ring samples taken from a control tube incubated as above and obtained by mixing ImI 0.1% peptone water and ImI a 10 -4 dilution of the collected saliva. The average colony count in the control multiplied by the dilution factor represents the initial inoculation density of the test mixture. Since the test mixtures containing mouthwash use undiluted saliva, any count that gave the same number of colonies as the control indicates a 99, 9% kill of the original inoculation. The time it took to effect this 99.9% kill was recorded.

Table VI

link

ADC

(mm)
S. aureus

SDS

(mm)

S. aureus

BCT

(Min.) Saliva

N-meth \ i-N- (2-hydroxyethyl) -N- (2-hydroxydodecyD-; 0.02

N-benzyl ammonium chloride 0.05

N-methyl-N- (2-hydroxyethyl) -N- (2-hydroxytetra-0.01

decyl) -N-benzylammonium chloride 0.05

N-methyl-N- (2-hydroxyethyl) -N- (2-hydroxy-0.02

hexadecyD-N-benzylammonium chloride 0.05

Cetylpyridinium chloride and decamethylene 0.02 (4-aminochinaldinium) acetate (comparison) p *

3.5

4.5

1
1

1.5

4 2.7

1.3, (2 **

5 3

p * denotes partial inhibition of the "seed" without a clear inhibition zone:

C ** denotes complete inhibition below the disc with no inhibition zone.

N-methyl-N- (2-hydroxyethyl) -N- (2-hydroxydodecyl) -N-benzylammonium chloride in a mouthwash was compared with commercially available quaternary ammonium compounds in various Find out concentration levels through the above tests. The results presented in the following Table VII are summarized, show that at all concentration levels according to the invention Compound (Compound 5) gave larger zones of growth inhibition than the others tested connections. In addition, compound 5 shows at a concentration level of 0.04 "n increased activity in that it took much less time to complete a 99.9 "oige To achieve killing of the salivary bacteria. The connection has the further advantage that the Mouthwash in which it is contained. does not have an unpleasant taste.

Table VII

10

Active ingredient

ADC zone size (mm)

S. aureus

St. faecalis

99.9% kill

Saliva (min.)

Control (no qualifying connection

1. Cetyl pyridinium chloride

2. Cetyldimethylbenzylammonium chloride

3. Cetylethyldimethylammonium bromide

4. Alkyldimethylnaphthylmethylammonium chloride * *)

5. N-methyl-N- (2-hydroxyethyl) -N- (2-hydroxydodecyl) -N-benzylammonium chloride (according to the invention)

**) 12 C 98%; 14 C 2%.

0.02 4.2 0.03 3.9 0.04 4.7 0.02 3.3 0.03 3.9 0.04 3.8 0.02 5.9 0.03 5.6 0.04 6.2 0.02 6.1 0.03 7.7 0.04 7.6 0.02 11.8 0.03 14.3 0.04 15th

3.2

3.4

1.9
2.6

4.3

4.7

3.1

3.4

4th
5
6th

V ₂

¹ Iz

³ U

Va

¹ U

¹ A ¹ U 1

V ₄

Vw (5 sec.)

The new quaternary ammonium compounds according to the invention can easily be incorporated into liquid, pasty or powdery agents, in which their germicidal properties can be used to advantage. Typical types of application for this Means are mouthwash concentrates, toothpastes, tooth cleaning agents and aqueous concentrates for Cleaning of food processing machines. The average content of the quaternary Ammonium compound in such agents can vary between about 1 and 5000 ppm. A a particularly suitable concentration range is between 200 and 2500 ppm.

An example of a suitable mouthwash concentrate is given below.

N-methyl-N- (2-hydroxyethyl) -N- (2-hydroxydodecyl) -

N-benzyl-ammonium chloride .. 0.100%

Sorbitol. 70% solution 20,000%

Cinnamon oil 0.050%

Peppermint oil 0.100%

Citric acid 0.100%

F. D. and C. Rid No. 2 0.001%

Polyoxyethylene sorbitan

Monostearate 0.300%

Ethyl alcohol 10,000 «/ ,.

Water 69,349%

. . This concentrate should be diluted with 3 to parts of water before use.

Claims (1)

Claim: Germicidal agent characterized by containing a quaternary Ammonium halide of the general formula R ¹ R ³ • CHOH CH ₇ -NR ² in which R ^{1 is} a C1 to C3 alkyl group or a C2 to Ce hydroxyalkyl group, R ^{2 is} a Q to Ca alkyl group or a C2 to C3 hydroxyalkyl group, R ^{3 is} a Ce to Cis alkyl group, X is hydrogen or halogen and Y is a halogen ion, as a germicidal agent. Considered publications: Swiss patent specifications No. 301409,303 510, 514. 809 588/446 7.68 O Bundesdruckerei Berlin