Products and method of decontaminating prions

FIELD: chemistry.

SUBSTANCE: group of inventions relates to a method of decontaminating products or materials infected with prions, and is intended for decontaminating medical or medical surgical devices at risk, decontaminating work surfaces, decontaminating animal meal or other contaminated products of animal origin. The step for decontaminating products or materials infected with pathogenic agents responsible for prion diseases comprises placing them in contact with a solution containing copper (Cu) and/or derivatives thereof. Said copper and/or derivatives thereof are present in an amount of at least 500 mcM.

EFFECT: use of the group of inventions improves efficiency of decontaminating infected products or materials.

7 cl, 4 ex

 

The object of the present invention are products and methods for disinfecting products and materials infected with non-standard vector-borne pathogens (ATNC) responsible for transmissible subacute spongiform encephalopathy.

The invention concerns, in particular, disinfection products and materials infected with prions, which accumulate mainly in the brain of the host, in particular in the form of abnormal isoforms of PrPSC(PrPscrapie), which is the result of a conformational change of the protein PrPC(cellular PrP), encoded by the owner.

The emergence of a new variant of the disease of Creutzfeldt-Jakob disease in the UK and its possible relationship with spongiform encephalopathy of cattle were brought to the fore the potential transmissibility of the disease, in particular, through an infected animal products, in particular, through contaminated food and medical-surgical supplies.

Indeed, modern methods of disinfection and sanitation cannot easily destroy prions, which are resistant to most commonly used methods. Administrative Guidelines prescribed rules that must be followed in this matter, but they are very difficult to implement, are expensive and require the use of toxic products.

In light of this the th context, the authors conducted a search of compounds, which would simply and efficiently to carry out the clearance of the pathogenic agent and which could be used on the vast majority of medical-surgical surfaces and materials. Their work showed the effectiveness in this regard, some metal derivatives.

The invention aims, therefore, the use of such compounds for disinfection of prion. It is also concerned with a method of handling infected materials and products, including the use of these compounds.

The invention relates, therefore, application of Cu and its derivatives for the treatment of prion.

According to the supplementary regulations of the invention, these derivatives are used in conjunction with H2O2.

Preferably the metal is derived CuSO4. One or more compounds used according to the invention, are mostly in the form of aqueous solutions.

Studying the effects of these derivatives on the clearance of pathogenic agents responsible for prion diseases, showed their high efficiency for purification of contaminated foods or materials.

Preferably these compounds are non-toxic, biodegradable, easy to use.

The invention relates thus also to the way abyssaria the Oia products or materials infected with pathogenic agents responsible for prion diseases, characterized in that it comprises bringing them into contact with at least one compound, as defined above, or its containing solution, or, if necessary, with H2O2.

In a preferred method of carrying out the invention a derivative of metal is CuSO4.

Disinfecting treatment is carried out mainly with the use of a solution containing the specified connection is based at least 500 μmol, in particular from 500 to 1000 µmol.

H2O2when it is used, is present in the solution at a ratio of approximately 50 mmol.

Satisfactory results were obtained by exposure at ambient temperature for about 15-60 minutes, in particular for approximately 30 minutes

This or these compounds and their solutions are, therefore, of great interest within the hospital, where they allow, in particular, to produce disinfection of medical or medical-surgical devices-risk groups, such as materials reuse, such as endoscopes, probes (dialysis), as well as decontamination of work surfaces such as laboratory table or the floor, also included in the risk zones.

These connections or irectory also especially useful for disinfecting, for example, infectious material of cerebral origin and biological products, derived from carriers of infectious forms of the disease of Creutzfeldt-Jakob disease. The invention also concerns the use of these compounds and their solutions for cleaning products blood origin or biological material used in transplants.

Preferably these compounds and solutions have proven effective on the prion strain associated with ESB (transmissible spongiform encephalopathy of cattle, or mad cow disease). They are applicable, therefore, to any type of prion whatever its strain and origin of the pathogen.

They can also successfully be used in food applications, in particular, to neutralize any potentially infectious compounds and, in particular, animal flour or other products derived from infected animals.

Worthy of interest applications include disinfection of areas such as abattoirs, surfaces and devices at risk of becoming in contact with infectious agents. Can be called, for example, especially those who come from ruminant animals.

Other characteristics and advantages of the invention are given in the following examples, in which reference is made to Fig. 1 and 2, which represent, with the responsibility.

Example 1. The solution for disinfection of prion-containing CuSO4and H2O2

a) study of the effects of copper together with H2O2the destruction of PrPSCpresent in the homogenates of infected mouse brain.

Solutions CuSO4and H2O2in various concentrations were added to the samples, consisting of extracts from infected homogenate of rat brain, and left in contact for approximately 30 minutes at ambient temperature. Then the samples were placed on a gel of polyacrylamide SDS-PAGE after equalization of the concentrations of proteins and decomposition with proteinase K (PK) at a concentration of 1 mg PK 50 mg of protein. The presence of PrPSCdetected using Western blotting.

The obtained results are illustrated in Fig. 1. It is seen that at a concentration of 100 µmol CuSO4and 50 mmol of H2O2infected samples have decreased levels of PrPSC. At a concentration of 500 μmol CuSO4and 50 mmol of H2O2the content of PrPSCpresent in homogenates of infected, is not detectable by Western blot (lanes 7 and 8). At this concentration effect is enhanced by the action of H2O2.

When you use the higher doses CuSO4from 1 mmol to 10 mmol only one copper eliminates signal PrPSCand on babok H 2O2is no longer required.

These results were confirmed by repeating these tests in homogenates derived from the brain of mice infected with strain 22L prion. Similar results were obtained with homogenates derived from mouse strain Chandler and ESB, which suggests that the disinfectant effect is not dependent on the strain.

b) study of the infectivity of homogenates of infectious brain processed solutions with a high concentration of copper in vitro.

Homogenates of brain 22L were treated for about 30 min with different concentrations of solutions CuSO4associated or not with H2O2in various concentrations. These infectious homogenates were subjected to dialysis, then was placed on cells of mouse neuroblastoma having the ability to fight the pathogen.

Homogenates of untreated brain were used as the control.

Western blotting was carried out after 6 passages in cell lysates and after decomposition with PK in order to detect the presence of PrPSCmeaning that the tested samples are still infectious.

The results are shown in Fig. 2. It can be stated that the infection is reduced as a result of processing.

Example 2. The in vivo tests

It was confirmed results observed in vitro. The brain homogenates infected with 22L, were treated, in particular, a solution of 500 µmol CuSO4and 100 mmol H2O2. The homogenates treated or not, were inoculated mice intracerebrally way. Control animals infected untreated homogenates, all sick for 168 days +/-2 days. Some animals infected-treated homogenates were alive even after more than 300 days. This result proves that the reduction of the titer of infection for at least 7 orders of magnitude can be obtained by this method of disinfection.

Example 3. Decontamination of surgical materials

Such materials, such as endoscopes, immersed in a solution containing l mg CuSO4on 20 minutes the Tests performed to identify the presence of prion after this treatment turned out to be negative.

Example 4. The effectiveness of disinfection

The following are data regarding the effectiveness of disinfection tested in vivo after vaccination in mice. For this control and disinfected the brain homogenates were inoculated intracerebrally way mice (C57B1).

The survival time of mice has the following values:

1) Inoculation control of infectious homogenates: 167,6 +/-0,5 days.

2) the Homogenates treated in CuSO4(0.5 mmol, 30 min, ambient temperature): 200,2 +/8.9bn days.

3) Homoge the ATA, processed in CuSO4(1 mmol, 30 min, ambient temperature): 217,2 +/-13,4 days.

4) Homogenates treated in CuSO4(1 mmol) + H2O2(100 mmol, 30 min, ambient temperature): 266,4 +/-15,6 days.

Taking into account such terms incubation periods in these different conditions and curve of the titration of the pathogen collected from these mice, it is possible to estimate that the achieved disinfection exceed 104in homogenates treated only CuSO4and above 105in homogenates treated in CuSO4+ H2O2.

1. The method of disinfection products or materials infected with pathogenic agents responsible for prion diseases, characterized in that stage decontamination is their introduction to the contact:
the solution containing copper (Cu) and/or its derivatives, and the copper and/or its derivatives are present in amounts of at least 500 microns.

2. The method according to p. 1, characterized in that the Cu and/or its derivatives are present in amounts of from 500 to 1000 microns.

3. The method according to p. 1, characterized in that the Cu and/or its derivatives are used in conjunction with H2O2.

4. The method according to p. 3, characterized in that the H2O2is present in the amount of approximately 50 mm.

5. The method according to p. 1, characterized in that a derivative of copper is CuSO .

6. Application of the method according to one of paragraphs.1-5 for disinfection of medical or medical-surgical devices-risk groups, such as materials reuse, such as endoscopes, or for decontamination of work surfaces such as laboratory table or the floor.

7. Application of the method according to one of paragraphs.1-5 for disinfection of animal flour or other contaminated foods of animal origin.



 

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13 cl, 3 ex, 4 tbl

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5 cl, 2 tbl, 2 ex

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3 tbl, 1 ex

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1 tbl

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