DE2402676C2 - Agents inhibiting the growth of microorganisms - Google Patents

Description

The invention relates to new agents which are used to inhibit the growth and kill microorganisms are suitable. The agents contain an ar- (halomethyl) -mandeic acid trile of the general formula

(D (NO ₂ ) ,, CN

in which X is a chlorine or bromine atom, m is 0, 1 or 2 and η 0, 1 or 2, as active ingredient in addition to customary carriers and / or diluents.
y> Typical examples of compounds of the general formula I are:

jr- (chloromethyl) -mandelsSurenltrtl

2-nitro-ar- (chloromethyl) -mandelic acid nitrll

4-Nltro-x- (chloromethyl) -mandelic acid trile
- ¹ ? 2 -ChloM-nltro-sr-dichloromethyD-almond acid ltrile

4-chloro-3-nltro-jr- (chloromethyl) -mandelic acid ltrll

2,4-dichloro-5-nltro-a- (chloromethy1) -nandelic acid nitrile

3-Nltro-ar- (chloromethyl) -mandels8urenitrll

2,5-Dlchlor-α- (chloromethyl) -mandelic acid ltrll
«Ο 3-Nltro- (z- (bromomethyl) -mandelic acid ltrll

2,5-dichloro-ar- (bromomethyl) -mandelic acid trile

^ - (ChlormethyD-Mandelsäurenltrll is a new compound within the scope of the present invention.

2,4-Dlchlor-jr- (chloro- ■ »< methyD-mandelic acid, 2- (chloromethyl) -mandelic acid and especially 3-nitro-cr- (chloromethyl) -mandelic acid.

The compounds contained in the agents according to the invention can easily and advantageously through Implementation of the appropriately substituted acetophenone with hydrogen cyanide (HCN) or an alkali or Alkaline earth salt thereof, conveniently in the presence of a mineral acid, can be synthesized.

*) The 7-HalogenmeihylmandelsäurenitrlIe can be used to kill certain microorganisms or stunt your growth. The classes of microorganisms are gram-positive and acid-fast Bacteria, yeast and fungi are particularly sensitive to treatment.

The microbiological activity of these α-halomethylmandelic acid bottles makes them particularly suitable for Use as anti-sludge additives for paints, as wake-up residue gels or germicides ίί Agent to prevent the formation of slime and algae in industrial waters and to prevent the formation of slime In cutting oils.

The synthesis of the ar- <halogenmetriyl) -mandelic acids contained in the agents according to the invention is carried out explained in more detail by the following production examples. In doing so, the structure of all manufactured products confirmed the nuclear magnetic resonance spectrum and the infrared absorption spectrum by elemental analysis. 1.0

Manufacturing example 1

100 g of 2-chloro-3-nitroacetophenone and 73.5 g of sodium cyanide were added to a mixture of 500 μl with stirring

cm 'of methylene chloride and 250 cm ^{1 of} water. The slurry was kept at 10 ° to 15 ° C. while 135 cm 'of concentrated hydrochloric acid was added dropwise over a period of 2.5 hours. The organic phase was then separated from the aqueous phase. The aqueous phase was washed with methylene chloride and that

Combined methylene chloride with the organic phase. This solution was washed with water, dried with magnesium sulfate and evaporated. 138 g of a crude product were obtained. This product was dissolved in methylene chloride, diluted with 10% hexane and cooled. 85 g of 3-nitro-st- (chloromeihyD-mandelic acid nitrile) were obtained Mp. 107-109 ° C.

Example 2

To a mixture of 26 g of 2-chloroacetophenone and 5 drops of a saturated aqueous solution of potassium cyanide 30 g of hydrogen cyanide were quickly added using a dropping funnel. The mixture became Heated to 34 ° C for 45 min. Excess hydrogen cyanide was absorbed in potassium hydroxide and the reaction mixture evaporated to give 25 g of a brown solid. The solid became from a 4: 1 mixture of hexane and benzene (activated charcoal) recrystallized. 17 g of ar- (chloromethyl) -mandanic acid nitrile were obtained Mp. 59 ° to 60 ° C.

Example i ι *

Following the procedure of Example 2, 2,4-dichloro- <z- (chloromethyl) -mandelic acid nitride, melting point 80 to 83 ° C. In 95% long yield from 2-chloro-2'4'-dichloroacetophenone and hydrogen cyanide. The following example shows the preparation and activity of a typical agent according to the invention.

Example 4

An aerosol formulation was made. By placing a Wheaton bottle with 100 g of the following mixture was filled:

0.1 «3-Nltro-« - (chlorometh / I) -mandelonitrile

59.9% stabilized methylene chloride

20% trichloroethanes

20% of a mixture of propane and butane

The effectiveness of the agents according to the invention was examined as follows.

Attempt 1

Typical a- (halomethyl) -manrielsäurenitrlle were tested for their microbiological activity against a variety of ^ number of microorganisms examined. In Table I are the fungistatic and bacteriostatic endpoints specified for certain bacteria, fungi and yeasts. For comparison are also data showing the activity of 2-phenyl-phenol and 4-Nltrophenol and 2,4-Dlchloratropanltrll and 0,2,4-Trichloratropanltril (US-PS 35 13 186) indicate has been included.

The test compounds were suspended in acetone or dissolved to prepare a concentrated solution * o. The final application concentrate ions were prepared by diluting this concentrated solution accordingly. The initial solutions were added to trypsin-containing soy broth in culture glasses. The final concentration of acetone was kept at 1% or below in order to prevent an adverse influence on the organisms. The culture glasses containing the media and chemical dilutions were then inoculated with the various test organisms. After an incubation period of 24 hours for the bacteria and 72 hours for the fungi, the presence or absence of growth of the organisms was observed. The numbers given in Table I represent the lowest dilution in parts per million (ppm) that prevented the test organisms from growing.

Table I. Fungistatic and bacteriostatic endpoints of α-halomethyl mandelonitriles Test Organisms Growth Inhibition Endpoint (ppm)

1 2 3 A B CD * '

Gram positive bacteria Staphylococcus aureus (Smith) Staph. aureus (FDA 209) Staph. aureus (Page)

Diplococcus pneumoniae Ι! Bacillus subtilis.

fi Streotococcus faecalis

4- 8 ~ 2.5 100-500 -100 32 32 10 4- 8 <32 32 32 2-4 <32 32 32 2-4 <32 > 25 10 8-16 5-10 > 50 32 8th 20th 8-16 <32 64 > 128

i-
j continuation 24 02 5 1- 2 616 > 25 B C 32 -500 D. 32 ■; Test organisms <32 > 25 Listeria monocytogencs Growth Inhibition 1
1 2 <32 ; 5 Corynebacterium equi Clostridium sporogenes 16-32 End point (ppm)
3 A 32 32 ι Ι " Gram negative bacteria 20th > 128 <32 Erwinia carotovora <32 -100 128 > 128 Escherichia coli (strain A) 5-10 > 128 > 128 100-500 > 128 15th E. coli (strain B) 50 > 128 > 128 > 128 E. coli (ATCC 9637) 100 40 > 128 <32 > Ϊ28 > 128 Salmonella typhimurium 50 > 128 25-50 > 128 > 128 2 (1 S. schottmuelleri '.- 5 25-50 100-500> 128 > 128 S. pullorum 5-10 > 128 > 128 S. choleraesius 40 > 128 1G-25 50-100 > 128 > 128 25th Serratia marcescens <32 > 128 > 128 Klebsieila pneumoniae > 128 <32 > 128 > 128 Proteus mirabilis > 128 10-25 > 128 > 128 Proteus vulgaris > 128 <32 > 50 64 > 128 JO Pseudomonas aeruginosa 8-16 <32 Schigella flexneri
Acid-fast bacteria 50-100 16-32 <32 35 Mycobacterium avium 16-32 <32 25-100 Myco ^ acterium smegmatis <32 40 Mycobacterium phlei 8-16 <ß_2 10-25 Yeasts 8-16 <32 Candida albicans <32 25-50 45 Saccharomyces cerevisiae 4- 8 <32 Mould 10-25 Aspergillus niger 1- 2 <32 50 Aspergillus fiavus <32 2.4-5 Penicillium citrinum Penicillium italicum 1- 2 5 -10 Penicillium variabile <32 10 -25 Epidermaphyton floccosum 2-4 * - 5 Trichophyton metnagrophytes <32 <32 St. I Trichophyton tonsurans <32 5 -10 Aspergillus tamari <32 <32 Chaetomiuiti slobosum <32
ά 5 »10 Hi Cladosporium res; iiae Fusarium moniliform <32 <32 <32 <32

continuation Test organisms

Growth Inhibition End Point (ppm) 1 2 3 A

Hormodendrum sp. Mcmnoniella echinata Microsporum gypseum Polyporus tulipiferus Poria mop.ticola Sclerotium rolfsii Trichoderma sp.

50

< 32 <32 < 32 <32 < 32 <32 < 32 10-25 < 32 <32 < 32 <32 < 32 <32

25-50

inventive means with

1- 2,4-dichloro-α- (chloromethyl) -mandelic acid nitrile

2 · a-fChlormelhyD-mandelic acid nitrile

3 - .l-Nilro-o-lchlormelhyD-mandelääurenilril

Comparison compounds A - 2-phenylphenol B * ■ 4-nilrophenol C «2.4-dichloro-atroponitrile D »^ .2.4-Trichloro-atropanitrile '

attempt

In order to investigate the killing effect of 3-Nltro-st- (chloromethyl) -mandels8urenltrll, a series of 18 mm cover glasses were sterilized and ^{inoculated with 0.01 cm 1 of} a 24 h old bacterial culture or a standard spore suspension of molds. As soon as the inoculum had dried, the cover glasses were placed in an aqueous solution of the compound to be examined for 10 minutes or 2 hours. The solution was drained from the coverslips which were then placed in jars containing sterile nutrient broth and incubated there to determine the viability of the organisms. The results of this study are given in Table II. The effectiveness of 2-phenylphenol in this experiment is also given for comparison purposes.

Table II Deadly effects of mandelonitriles on microorganisms link Test organisms

Dose (ppm) required to kill 10 minutes, 2 hours

3-nitro-a- (chloromethyl) -

almond nitrile

2-phenylphenol

Aspergülus niger Penicillium variabile Staphylococcus aureus Salmonella choleraesius Mycobacterium avium

Aspergülus niger Penicillium variabile Staphylococcus aureus Salmonella choleraesius Mycobacterium avium

attempt

Often times, compounds are less effective when applied in the presence of organic substances will. A standard attempt to determine the increased dose that is required. if organic

<250 <250 > 1000 <250 > 1000 > 1000 > 1000 > 1000 250-500 ~ 500 250-500 250-500 250-500 250-500 250-500 250-500 <250 <250 <250 <250

Substances are present. Is the serum exposure test. This is basically the Im Example 3 illustrated static glass thinning tests with a range of concentrations of the tested Compound In Trypsin-Containing Soy Broth And A Similar Series Of Dilutions In The Same Medium additionally containing 5 * sterile horse serum. The difference in the growth inhibition * The same concentration in the media with and without serum gives an approximation for the increased dose that In Presence of organic substances is required. Such an attempt is summarized as follows:

Table St.
, "Serum inactivation of 3-nitro-α- (chloromethyl) -nandelonitrile

Minimum inhibitory dose (ppm)
Organism with horse serum without horse serum

Staphylococcus aureus 25-50 2.5-5

The agents according to the invention contain a carrier and / or a diluent, the inert being The carrier can be a liquid or a solid. Liquid carriers in which the «- (halomethyl) -mandelic acid ltrll Present in solution are water and organic solvents such as alcohols, ketones, allphatlsche

^{: o} and aromatic hydrocarbons and halogenated aliphatic and aromatic hydrocarbons. Suitable surfactants for suspending or emulsifying the ate- (halomethyl) -mandelic acid parts with the liquid carrier can also be added. Solid carriers such as talc, bentonite, kieselguhr, etc. can also be mixed with the ar- (halomethyl) -mandelic acid oils to produce the agents according to the invention.

^J > The concentration of cr- (halomethyl) -mandelic acid oil required to inhibit the growth of microorganisms varies with the specific type of α- (halomethyl) -mandelic acid oil, the type of microorganisms, whether a carrier is used or not, the environmental conditions, etc. However, the person skilled in the art can easily determine an appropriate concentration for the respective area of application, e.g. B. by making comparisons. When the «- (halomethyl) -mandelic acid bottles with a carrier

"'" are mixed, the concentration of active ingredients is usually in the range of about 10 to about 1000 parts per million, and preferably from 10 to 50 ppm.

Attempt 4

M The lethal effect of the aerosol according to Example 4 on microorganisms was investigated. By inoculating an inch (2.54 cm) square of cheesecloth with three bacteria and two types of fungus. The inoculated rectangles were sprayed with the aerosols for 3 seconds, slowly walking three times over the inoculated pieces of cheese linen. After this treatment, the pieces of cloth were placed in 1-liter bottles, which were tightly closed and incubated for 24 and 48 hours, respectively. After these two tents, the cloth

Put 4n pieces in jars containing a sterile soy broth containing irypsin in order to ensure the viability of the organ nlsmen to determine. The results of such an experiment are summarized in Table IV.

Table IV
J ₅ Lethal effect of the aerosol spray on microorganisms

Number of attempts with no growth
occurred ')

Organisms 24 h 48 h Staphylococcus aureus 5 8 Escherichia coli 5 8 Mycobacterium avium 2 2 Aspergillus niger 10 10 PeniciUium variabile 10 10

⁴ I 10 total repetitions for each attempt

Claims (1)

Patent claims: 1. The growth of microorganisms inhibiting agent, containing a üc- (HalogenmethyI) -mandeläureniltril of the formula _- - in which X is a chlorine or bromine atom, m is 0, 1 or 2 and π is 0, 1 or 2, as active ingredient in addition to customary carriers and diluents.
2. x-iChlormethyO-mandelic acid nitrile.