DE1204777B - Synergistic potentiation of hexachlorophene - Google Patents

Description

FEDERAL REPUBLIC OF GERMANY

GERMAN

PATENT OFFICE

EDITORIAL

Int. Cl .:

Number:
File number:
Registration date:
Display day:

A611

German class: 3Oi-3

St 23651IV a / 30 i

April 8, 1965

November 11, 1965

The present invention relates to the potentiation of disinfecting or germicidal and fungicidal Effect of hexachlorophene by means of unsubstituted aromatic amines, amides and acids, which usually have little or no disinfectant or germicidal or fungicidal properties. In particular, it relates to the potentiating effect on hexachlorophene in acidic media by adding an aromatic compound of the formula

Synergistic potentiation of hexachlorophene

in which R denotes a group —COOH, —NH ₂ or - CONH ₂ .

Hexachlorophene is the common name for 2,2'-methyl-bis- (3,4,6-trichlorophenol), the Widely sold as a disinfectant and fungicide. According to the invention, the effectiveness this compound is increased significantly by adding one or more of the compounds: benzoic acid, Aniline and benzamide, in a weakly acidic solution with a pH in the range of about 6.0 to about 6.8 is added.

The following examples explain the results achieved according to the invention:

example 1

The following dilutions were made in 1000 ml of brain-heart infusion fluid, their pH 6.8 was:

A. Hexachlorophene 10 mg

B. Hexachlorophene 10 mg

and benzamide 10 mg

C. Hexachlorophene 10 mg

and benzamide 50 mg

D. Hexachlorophene 10 mg

and benzamide 100 mg

E. Benzamide 10 mg

F. Benzamide 50 mg

G. Benzamide 100 mg

These preparations were further diluted in 10 ml quantities in sterile culture test tubes produced in the range of 1: 100,000 to 1: 9,000,000 hexachlorophene in strength and from 1: 10,000 to 1: 9,000,000 benzamide. All dilutions made in this way were inoculated with 0.1 ml of a 24 hour old culture broth of Staphylococcus aureus and Incubated for 48 hours at 37 ° C.

Applicant:

Dr. Herbert C. Connector,

Ho-Ho-Kus, N. J. (V.St. A.)

Representative:

Dr. W. Kühl, patent attorney,

Hamburg 36, Esplanade 36 a

Named as inventor:
Dr. Herbert C. Connector,
Ho-Ho-Kus, NJ (V. St. A.)

Upon subsequent testing, the highest dilution of non-turbid agent (i.e. Bacterial growth), selected as the final product, and this represented the minimum inhibitory concentration (MIC). The results obtained were as follows:

35 middle 10 mg MIC A. Hexachlorophene 10 mg 1: 2,500,000 B. hexachlorophene 10 mg and benzamide 10 mg 1: 3,425,000 40 C. Hexachlorophene 50 mg and benzamide 10 mg 1: 3,425,000 D. Hexachlorophene 100 mg and benzamide 100 mg 1: 5000,000 E. benzamide worse than 45 1: 10,000

From the above results it can be seen that the addition of benzamide in one concentration from 50 to 91 percent by weight of the mixture with hexachlorophene an increase in germicidal Effectiveness around 37 to 100% results. Synergistic effects of benzamide have been shown in concentration

509 737/341

Trations of only 10% benzamide in the mixture and up to 95% were observed.

Example 2

In order to determine the effect of the agents according to the invention with regard to the killing time, made the following dilutions at pH 6.8:

H. Hexachlorophene l% ige suspension I. Hexachlorophene 1% and benzamide 1% J. Hexachlorophene 1% and benzamide 5% K. Hexachlorophene 1% and benzamide ΙΟ «» / ,, L. Benzamide l% ige solution M. Benzamide 5% suspension N. benzamide 10% suspension

From each of these dilutions were in sterile test tubes in an amount of 5 ml further dilutions were made, which were in their strength in the range from undiluted to 1: 1000.

All dilutions made in this way were made with 0.5 ml of a 24 hour old Staphylococcus culture broth aureus inoculates. At precise intervals of 5 minutes, 10 minutes and 15 minutes an amount of a 4 mm loop was completely removed and continued in sterile brain-heart infusion fluid bred. After shaking, an amount of 4 mm loop was fully grown further. This precaution proved necessary to avoid false readings resulting from a possible transfer of the remaining inhibitory effect of hexachlorophene in the first subculture glasses could result.

After incubation at 37 ° C. for 48 hours, all subcultures were checked for growth. The dilution of any agent that kills the test organisms in 10 minutes, but not in 5 minutes was determined as the end point. The results were as follows:

Example 3

To determine the effect of pH on. the effectiveness of hexachlorophene in the presence of benzoic acid, the following solutions were made in bovine heart infusion fluids:

O. Hexachlorophene (5 mg in 500 ml) pH 7.4

P. hexachlorophene, 5 mg,

ίο and benzoic acid, 0.5 mg (5 mg in 500 ml) pH 7.4 Q. Hexachlorophene (5 mg in 500 ml) pH 6.5

R. hexachlorophene, 5 mg,

and benzoic acid, 0.5 mg (5 mg in 500 ml) pH 6.5

Twenty-six dilutions of each solution were made, their concentrations between 1: 1,000,000 and 1: 100,000,000 were. A control broth of each pH was made with Staphylococcus aureus inoculated to determine any difference in growth. The minimum inhibitory concentration for each solution was found as follows:

1% Dilution to kill middle 1% in 10 minutes, however 1% not in 5 minutes A. Hexachlorophene 1% 1: 150 I. Hexachlorophene 10% Benzamide 1% 1: 175 J. Hexachlorophene 10% Benzamide 1% 1: 175 K. Hexachlorophene 10% Benzamide 1: 250 L. benzamide, no N. Benzamide, not to kill in 15 minutes empowered

From <len previous results one can conclude that benzamide accelerates the killing power of hexachlorophene, especially in the ratio of 1:10 (as in experiment K.), with the increase in effectiveness in the vicinity of 67%.

O. middle pH 7.4 1: 1 MIC 25th P. Hexachlorophene pH 7.4 1: 1 840,000 Q- Hexachlorophene pH 6.5 1: 2 30 R. and benzoic acid 660,000 Hexachlorophene pH 6.5 1: 2 100,000 Hexachlorophene and benzoic acid 450,000

From the above results it can be concluded that benzoic acid sees in a weak alkali Medium acts as an antagonist and actually lowers the MIC. In weakly acidic solution Benzoic acid acts (in a ratio of 10%) as a potentiator, whereby the effectiveness of the Hexachlorophene is increased by at least 17%.

Example 4

The inhibiting agent (given below) was dissolved in 1000 ml of nutrient broth adjusted to pH 6.5 had been. This dilution became 10 ml of dilutions in sterile broth in the starch range between 1: 100,000 and 1: 4,000,000 (20 test tubes). Any dilution was inoculated with 0.1 ml of a 24 hour old Staphylococcus aureus culture broth. At all The tube density was read in a densitometer and the values recorded. A control broth was inoculated. All tubes were incubated at 37 ° C for exactly 24 hours.

After the incubation, the density in all tubes was determined again in the densitometer, since the Growth causes opacity; the higher the reading in the densimeter, the larger the lower the growth and vice versa

the effectiveness of the agent. A reading in

the densimeter, which is the same as before the incubation; is, shows no growth and therefore one 100% effectiveness of the agent.

The broths were chosen to have an increase

the reading in the densitometer of 0.02%, the reading before the incubation and showed who were thus those in whom growth first appeared; this was considered the end point.

Based on the aforementioned feature, the following results were obtained:

S. hexachlorophene 10 mg 1 : 2,500,000 T. hexachlorophene 10 mg and benzoic acid 100 mg 1 :> 4000 000 U. hexachlorophene 10 mg and aniline 100 mg 1 :> 4000000 V. Benzoic acid 100 mg 1 : <10,000 N. aniline 100 mg 1 : <10,000

The above results show that both benzoic acid and aniline act as potent potentiating agents Agent for hexachlorophene acts, the concentrations showing such an effect, be in the range of 10 to 95 percent by weight of the composition in hexachlorophene.

The following compounds tested were found to be either potentiating Agents are ineffective or act as antagonists for hexachlorophene: salicylic acid, salicylamide, Stearylamine, piperazine, benzanilide, pyridine, benzene, Dimethylaniline, alcohol, salicylanilide, p-aminobenzoic acid and many others.

The tests were carried out here using Staphylococcus aureus as a breeding organism, but these have turned out to be potentiating compounds described have also been shown to be effective against both gram-negative and gram-positive bacteria, including E. Coli.

The potentiating effect of the aromatic compounds mentioned above has been proven in amounts of from about 1 to about 5 percent by weight and up to 75 percent by weight and higher on the potentiated mixture, proved to be detectable.

Claims (2)

Patent claims: 1. Synergistically potentiated bactericide, characterized by a content of Hexachlorophene and about 1 to about 95 weight percent of a compound of the formula in which R denotes a radical —COOH, —NH ₂ or —CONH2, in an acidic medium. 2. Synergistically potentiated bactericide according to claim 1, characterized in that it has a pH of 6.0 to 6.8. 509 737/341 11.65 © Bundesdruckerei Berlin