DD299418A5 - DISINFECTANT MEASUREMENT FOR THE TARGETED INACTIVATION OF GERMINES - Google Patents

Abstract

The invention relates to a disinfectant solution for disinfecting and disinfecting cleaning. Carboxymethylaminomethylsulfone-pyrrolidinium-sulfomethyl-betaines of the formula (I) are used, which are significantly synergistically enhanced by alcohol in its disinfecting action. According to the invention, a comprehensive germ-reducing effect against viruses, bacteria, fungi, parasitic protozoa and helmints is achieved, which can be used both for skin disinfection and for disinfecting smooth and rough surfaces. In contrast to peracetic acid there is no weakening by blood. Conventional disinfectants with acidic, oxidative, irritating, sensitizing or corrosive properties can be replaced. Formula (I) {Degradation; Disinfection; disinfecting cleaning; synergistic effect; sulfobetaines; sulfone; viruses; bacteria; mushrooms; parasitic protozoa; Pyrrolidinium betaine; Amino acids}

Description

HOOC-CH ₂

takes place, which can be used in water in concentrations of 0.01 to 2 wt .-% or for the synergistic increase in activity in the same concentration in water / alcohol mixtures with a share of 20 to 99 vol .-% water and 1 to 80 vol. % Alcohol to be solved.

2. disinfectant solution according to claim 1, characterized in that are used as the alcohol, ethanol or a propanol.

3. disinfectant solution according to claim 1 and 2, characterized in that for skin disinfection with specific activity against hepatitis B viruses, the active compounds of the formulas (I) and (II) in water / alcohol mixtures having an alcohol content of 30 to 80 vol. -% to be solved.

4. disinfectant solution according to claim 1, characterized in that for the complete dissolution of blood contaminants, the active compounds of the formulas (I) and (II) in the concentration range of 0.1 to 2 wt .-% are used.

Field of application of the invention

The application relates to the germ killing / inactivation of germs having a broad effect in the human and veterinary field as well as in the food industry.

The application is possible for facilities where measures for a multivalent germ reduction, a germ-reducing cleaning or disinfection are necessary.

Characteristic of the known state of the art

Multivalent agents for killing germs, which are characterized by user-friendly wedge, high efficiency as well as by toxicological and ecological criteria, take into account the capillary problem ui. And last but not least economically feasible, do not yet exist. Due to the diversity of germs, the fields of application, products and processes or the compatibility and environmental impact of disinfectants, the area is very confusing and in need of improvement. A classification according to chemical substance classes with reference to the respective fields of application is also difficult. On the other hand, because of the diversity of the germs, the disinfectants used are also kept variable, and the chemical agents are often used in combination or together with physical agents (e.g., temperature elevation).

Because of the versatility of the application criteria, the individual disinfectants are therefore adapted for the individual disinfection procedures and especially disinfection tasks (hand, skin, instrument disinfection, etc.). The demands for faster and safer disinfection are difficult to reconcile η with the requirement for compatibility, the requirement not to appear corrosive and other environmental and economic aspects such as durability, shelf life, odor, skin irritation and handling. According to the prior art, high efficiency

and safety accompanied with disadvantages in handling. In the search for new, safe disinfectants antiviral MiUeI have not been dealt with extensively enough, whereby the above-mentioned disadvantages for virus disinfection in particular MaUe exist and the range of usable antivlal disinfectants is still small. In chemical germ killing the following means have been used so far (HORN, PRIVORA, WEUFFEN, "Handbook of Disinfection and Sterilization" Volumes 1 to 5, Publisher Volk and Health 1972 to: 984):

- aldehydes or mixtures of aldehydes;

Alcohols, especially ethanol and the propanols;

- halogen compounds, especially iodine compounds and hypochlorites;

- lactones;

- metal compounds (mercuric chloride, copper sulfate);

- Oxydants (peracetic acid, ozone), epoxides;

Phenols, xylenols;

- Quaternary ammonium compounds;

special surfactants, e.g. carbobetaines;

where new types of disinfectant have not yet prevailed.

Of the compounds mentioned, e.g. Peracetic acid and aldehydes a very wide range of action. These However, substances are generally toxic, unstable in storage and / or have high corrosivity. Add to that a quick- / Inactivation of peracetic acid by blood, resulting in a significant effect restriction. Formaldehyde leads due

its sensitizing effect on occupational diseases. Sporicidal preparations such as performic acid and ethylene oxide are especially toxic, e.g. classified as carcinogenic. Phenols have clear gaps in the spectrum of action and are usually unpleasant odor intensive. Hypochlorites are used irrespective of the significant user unfriendliness. The effect of alcohols against hepadnaviruses can not be established with certainty, therefore they are used in high risk areas with other drugs. Also exist in the previously used cationic surfactants and the Carbobetainen

Activity gaps. Effect against selected viruses has been demonstrated for:

- aldehydes (formaldehyde, glutaraldehyde);

- alcohols (ethanol, propanol);

- halogen compounds (especially hypochlorites);

- peracetic acid;

- some zwitterionic detergents.

In the evaluation of disinfectants, viruses and bacterial spores are a major criterion. As test viruses have been used very often; Influenza viruses, vaccinia viruses, poliomyelitis viruses;

then follow:

Coxsackieviruses, ECHO viruses, adenoviruses, herpesviruses. Influenza and vaccinia viruses are considered to be about the same in their behavior towards disinfectants-despite e.g.

different ether resistance.

Hepadnaviruses are considered to be particularly resistant. Object of the invention

The aim of the invention are those disinfectant solutions that allow a multivalent germ killing by chemical or physicochemical means in compliance with application criteria, namely simultaneously gegon Bacteria, fungi and viruses and against other germs, which achieved under application to different conditions of use safe germ killing and a toxic damage is counteracted by the user.

Explanation of the essence of the invention

The object, which is solved by the invention, is to find such disinfectant solutions by which a multivalent germination / inactivation of germs without reactivation of the germs is achieved. These disinfectant solutions are geared to

high activity against viruses, bacteria, spores and worm eggs;

- ease of use (little or no skin irritation, no corrosion effect, certain cleaning effect, low odor control, water dissimilarity);

- favorable characteristics (toxicological, carcinogenic, mutagenic, teratogenic and ecological criteria);

- Consideration of the capillary problem or penetration into pores or rough surfaces;

- dirt dissolution; Dissolution of blood admixtures;

- economic and raw material-side producibility.

The object is achieved according to the invention in that the germ reduction / disinfection by carboxymethylamino-methylsulfone-pyrrolidinium-sulfomethyl-betaines of the formulas (I) and (II),

HOOC-CH ₂ -NH-CH ₂ - £

Hi

HOOC-CH-NH-CH ₉ -So ₉ CH,

HOOC-CH ₂

which can be prepared very simply by heating 1,1-dimethyl-3-sulfinic acid-methyl-4-sulfomethyl-pyrrolidium-betaine (prepared according to DD PS 225128), formaldehyde and amino acid Ir water to 10O ⁰ C in aqueous solutions in total concentrations between 0 , 01 to 2Gew .-%, which may contain ethanol or a propanol for the synergistic increase in activity, is made.

In the concentration range of 0.1 to 2Gew .-% active ingredient complete dissolution of blood Vorschmutzungen is achieved. Surprisingly, it has been found that even aqueous solutions in low concentrations cause strong inactivation of various germs, while highly resistant germs (e.g., hepatitis B viruses) are inactivated by the combination of the polyfunctional sulfobetaines with alcohols (ethanol, propanols); Such a synergistic effect-increasing effect occurs according to the invention in solutions. which contain 20 to 99% by volume of water and 1 to 80% by volume of alcohol. Against hepatitis B viruses, 30 to 80% by volume of ethanol or propanol are used in combination with the sulfobetaines according to the invention. Through this favorable Kombinierbarkelt lit it possible to adapt the method for multivalent germ killing various application conditions. This also makes it possible to achieve specific effects against Bacierien, spores, fungi, viruses, parasitic protozoa and worm eggs.

The disinfectant solution according to the invention is suitable for skin disinfection as well as for reducing the germ on hard or rough surfaces, for example appliances, painted surfaces, metals, enamels and ceramics. The disinfectant solution according to the invention does not irritate the skin in the neutral region, does not corrode, shows no unpleasant odor, is non-toxic and can be stored indefinitely without any problems both as a substance and in solution. If necessary, it can be combined with other aqueous or aqueous-alcoholic agents. By the aforementioned property image, it is superior to conventional disinfectant solutions in effect and use value and can be produced inexpensively.

Ausführungsbelsplele example 1 To a microorganism mixture with 10 ⁹ germs / ml in strongly viscous nutrient medium from Bacillus subtilis, Endomycopsis

bfspora including their spores, the carboxymethyl-aminomethylsulfon-pyrrolidinium-sulfomethyl-betain the

Formula (I) in a final concentration of 0.2 wt .-% mixed and incubated for 30 minutes. After deactivation of the By dilution with sterile tap water, the test for sterility with the nutrient media is carried out in accordance with Pharmacopoeia Volume I, X, 4.0.1.ff. and 7.0.1. ff. Results:

- with the active substance of the formula (I): sterile

- Control without active ingredient: germination

Example 2 Chicken blood containing 10% by volume citrate buffer is injected with influenza virus type A strain A / PR / 8/34 in allantoic fluid

given. After thorough mixing, to this is added such a volume of a 10% solution (pH 7) of the carboxymethylaminomethylsulfone-pyrrolidinium-sulfomethyl-betaine of formula (II) in 10% ethanol that the final concentration of active ingredient is 0.25% by weight. After mixing again for a period of 10 minutes, it is deactivated by addition of at least 100 times the volume of a buffered NaCl solution.

Thus, pre-incubated chicken eggs are inoculated according to the usual methods for the propagation of influenza viruses. Of these Eggs are sterile removed after an incubation period of at least 48 hours Allantolsflüssigkeit and the Virus multiplication compared to control inoculated with NaCl solution and to the hatching eggs without active ingredient by means of the Hemagglutinatlon test comparatively assessed. Results:

- with active substance of formula (II): no virus replication

- without Wiikstoff: full virus multiplication

Inactivation testing was performed according to WHO requirements for Influenza Vaccine (Inactivated).

8elsplel3

To a buffered virus suspension containing at least 10 ⁴ infectious units per ml of adenovirus (human) type 2

such a volume of a 10% neutral solution of the carboxymethyl-arninomethyl-sulfon-pyrrolidinium sulfornetKylbetains of formula (I) in sterile tap water, that the final concentration of active ingredient is 0.4%.

After mixing and an exposure time of 30 minutes, the volume is increased by at least 100 times

deactivates a lactalbumin-containing phosphate buffer solution.

Thus, a permanent cell line suitable for the propagation of these adenoviruses is inoculated by the usual methods. From

Each culture vessel is subse- quently sub-cultured and viral replication compared to the unvaccinated control and to the cell cultures is evaluated with no adenovirus using the cytopathic effect (CPE) and immunofluorescence comparison.

Results:

with active substance of the formula (I): no CPE

- without active substance: CPE

Example 4 Inactivation of hepatitis B surface antigen (HBsAg) in the suspension test: Equal parts of a HBsAg-containing serum pool in various dilutions and the disinfectant to be tested

be mixed. After a reaction time of 30 minutes, the samples are inactivated by a dilution of 1: 100.

The residual activity of HBsAG is determined by radioimmunoassay compared to a control batch. Results: Restak · Control

tivity approach

= 100%

Serumpool serum pool

neat. 1:50

0.1% active ingredient of formula (I) 10% 8%

in 70% ethanol

0.1% in water 90% 65%

Example 5 Mäuseschwänzchentest

10 mm long tail pieces are inserted after thorough pre-cleaning in a HBsAg-containing serum pool for 5 minutes and dried for 30 minutes at room temperature. For the disinfection test, the tail pieces are individually placed in 50 ml measuring cups, the bottom of which is covered with glass beads, 1 ml of disinfectin solution (0.1% solution of the mixture of carboxymethylaminomethylsulphonopyrrolidine-sulfomethyl-betaine of the formula (I) in 50% n-propanol) and shaken for 2 or 30 minutes. Thereafter, the sample is diluted 1:50 and the residual activity of HBsAg with a

Radioimmunoassay determined. The tail pieces are covered with 3 ml of physiological saline and again

Shaken for 5 minutes. The residual activity in the rinsing liquid is again determined by a radioimmunoassay.

Results: Residual activity of HBsAg (control = 100%) Exposure time 2 min exposure time 30 min ' Disinfection Rinse · Disinfection Rinse Solution Solution Solution Solution

9% 2.5% 6% 1%

Example β

Guinea pig skin test Fur pieces that have been removed from shaved dorsal skin of guinea pigs become buttons with a flat surface

(2.5 cm in diameter), sutured on the back with surgical suture material and fixed by wooden sticks inserted in the buttonholes. For each test, 2 germ carriers prepared in this way are required.

A germ carrier is added with 0.05 ml of the disinfectant solution to be tested and r 'aid the second germ carrier

washing movements performed. After a contact time of 2 or 30 minutes, the disinfectant solution is washed off and then the germ carriers are rinsed for 5 minutes with 3 ml of 0.9% saline. The residual activity of the

HBsAg was determined by radioimmunoassay. Results:

0.1% solution of carboxymethylaminomethylsulfone-pyrrolidinium-sulfomethyl-betaine of the formula (I) in 90% strength n-propanol; Residual activity of HBsAg.

Control substance = 100% Exposure time 2 min exposure time 30 min

6% 2%

Example 7

To a virus suspension (virus-containing allantole fluid) whose content of influenza viruses (type A, WHO reference strain

A / PRS / 34 or Type B, WHO reference strain B / Arbor 1/86) is at least 10 'to 10 ⁹ ID _M / ml, such volume is a 10% solution of carboxymethylaminomethyl-sulfone-pyrrolinedin-sulfonethyl-betaine of the formula (I) In sterile tap water, that the final concentration of active ingredient is 0.1%. After mixing and a 16 minute exposure time, deactivation is effected by the addition of 100 volumes of HANKS medium.

Thus, according to the method known as propagation system for influenza virus method allantois-on shell about 5 x 5mm large egg shell pieces with chorioallantiosmembran (CAM) of 13- to 14 days embryonated chicken eggs in HANKS medium inoculated. After an incubation period of 48 hours (type A) or 72 hours (type B), the eggshell pieces are removed with the CAM and the virus multiplication in the HANKS medium is compared with the virus control without disinfectant by means of the hemagglutination test.

The toxicity of the disinfectant solution is investigated by a known method directly on the DAM. The virus multiplication after the disinfectant action is checked with non-toxic doses.

Results:

with active compound of the formula (I): reduction of the infectious agent by> 4 titer levels (log 10) without active ingredient: virus titer unchanged

Example 8

Preparation of Active Ingredients (I) and (II)

For the preparation of the new amino acid derivatives, the amino acid with!, I-dimethyl-S-sulfinsäuremethyM-sulfomethylpyrrolidinium betaine (available to DD PS 2251128) and formaldehyde in the molar ratio 1: 1: 1 by heating in water.

Example for the synthesis of the carboxymethylaminomethylsulfone-pyrrolidinium-sulfomethyl-betaine of the formula (I):

170.5 g (0.5 mol) of 80% sulfinic acid component are mixed in 1.41 with 50 g (0.5 mol) of 30% formalin solution, heated to 60 ° C and added 37.5 g (0.5 mol) of glycine. It is allowed to boil for 1.5 hours under gentle reflux and concentrated to dryness.

Schmp.112bis116 ⁰ C.

It is a colorless, readily water-soluble product.

The synthesis is analogously applicable to other amino acids.

Claims (1)

1. disinfectant solution for targeted inactivation of germs, characterized in that the germ reduction, germ-reducing cleaning and / or disinfection by carboxymethylsulfone-pyrrolidinium-sulfomethyl-betaines of the formulas (I) and (II) HOOC-CH ₂ -NH-CH ₂ - (D HOOC-CH-NH-CH ₉ -SO I ²