FR2509577A1 - METHOD, COMPOSITION AND PRODUCT FOR ACTING AGAINST VIRUSES - Google Patents

Abstract

THE INVENTION RELATES TO A METHOD, A COMPOSITION AND A PRODUCT FOR ACTING AGAINST HARMFUL RESPIRATORY VIRUSES. THE TECHNICAL PROBLEM IS TO PREVENT THE TRANSMISSION OF DISEASES. THE PRODUCT, ACCORDING TO THE INVENTION, INCLUDES ONE OR MORE ACIDS SUCH AS CITRIC, MALIC, SUCCINIC, BENZOIC ACIDS AND THE LIKE IN AN EFFECTIVE PROPORTION AND MAY ALSO CONTAIN A SURFACTANT. THE INVENTION IS APPLIED IN PARTICULAR TO THE TREATMENT OF RHINOVIRUSES, PARAINFLUENZAVIRUSES AND ADENOVIRUSES.

Description

1 - "Method, composition and product for acting against viruses"

  The present invention relates to virucidal compositions of high efficacy against viruses

  respiratory systems such as rhinoviruses, parain-

  The invention relates more particularly to a novel type of a virucidal composition that can be applied to a variety of substrates such as cellulosic tissues, nonwoven structures, and other materials. textile base In addition, the class of compositions

  virucides belonging to the present invention may also be

  incorporated in nasal sprays, face creams, hand lotions, pencils

  lips and other similar cosmetic preparations.

  can also be used as ingredients in kitchen and bathroom detergents, furniture and floor polish products and

household maintenance, similar.

  Virologists specializing in the field of respiratory viruses generally agree

  recognize that rhinoviruses, influenzaviruses and

  noviruses are among the largest group of pathogens

  responsible for respiratory diseases Rhinoviruses, in par-

  especially, are considered to be the main responsible

  which is commonly known as the "brain cold".

  The term "rhinovirus" that touches the nose, results in abundant runoff of the nose when the

  infections are due to this group of viruses The rhinovirus belongs to

  is due to the family of picornaviruses, viruses which, because of the absence of an outer envelope, are often characterized as "naked viruses" Although we know more than a hundred different antigenic types of rhinovirus, they have in common some important attributes For example, all are endowed with an ether-resistant capsid and all contain a "single chain" RNA (about 2.6 x 10 daltons) All are difficult to -2 inactivate by usual germicides such as compounds

quaternary ammonium.

  Adenoviruses have more than 30

  antigenic types When they enter the respiratory tract

  they cause inflammation of the tissues leading to symptoms of pharyngitis, bronchitis, etc. Although most adenovirus conditions occur during childhood, it is not uncommon for adults to also be affected by this type of infection. Like rhinoviruses, adenoviruses do not have an envelope, but these, unlike rhinoviruses, contain a double DNA chain Adenoviruses are

  usually resistant to inactivation.

  Parainfluenzaviruses belong to the paramyxovirus family They play an important role in the development of diseases of the lower respiratory tract

  in children and diseases of the upper respiratory tract

  in adults Parainfluenzaviruses are viruses

  containing RNA with a sensitive lipoprotein envelope.

  These viruses are resistant to inactivation by the carboxylic acids at the nucleocapsid.

low concentrations.

  A recent study by Dick et al. (Dick, EC and Chesney PJ "Textbook of Pediatric Diseases", Feigin, RD and Cherry, JD ed. Vol. I, p. 1,167 (1981) WB Saunders Co Pub Phila, PA) threw a new light on the mode of transmission of respiratory diseases caused by rhinoviruses Although the exact mode of transmission of respiratory diseases is not fully understood, studies carried out in the field by the above-mentioned researchers have shown that effective transmission diseases such as colds in the brain usually require that the subject being contaminated and the potential victim be very close to each other or in direct or indirect contact (indirect contact may be regarded as contact through 3- through an intervening surface, for example the top of a table, the latch of a door, etc.) Thus, it is possible to interrupt the chain of infection and reduce its tendency to spread if viruses can be rendered ineffective when they come out of the nose of an infected person by immediately exposing them to a virucidal agent In addition, once the viruses come out, those seeking refuge on the figure or on the hands of the infected person, can also be "killed" if a suitable virucidal agent is rapidly

  brought into contact with the appropriate anatomical surface, that is,

  to say the figure, the hands, etc. A facial tissue containing a composition acting quickly and having a virucidal action

  effective is a simple way to achieve this goal.

  We have long felt the need to.

  having a virucidal agent that is safe and effective against the usual respiratory viruses Commonly used simple germicides are ineffective against rhino and adenoviruses.

DESCRIPTION OF THE PRIOR ART

  U.S. Patent No. 4,045,364 to Richter discloses a paper impregnated with an iodophor (i.e., iodine and a carrier) having germicidal and useful properties as

  Prewash means in routine surgical operations.

  The patentee has discovered that the stability of the iodophor is increased at low pH and that small amounts of weak organic acids such as citric acid or acetic acid can be added to allow control of the iodophor. U.S. Patent No. 3,881,210 et al discloses a pre-moistened tampon for sanitary uses that may contain a bactericide US Pat. No. 3,654,165 to Bryant et al discloses a cleaning means for wiping, containing Bactericidal iodine imparting US Pat. No. 3,567,118 to Shepherd et al discloses a fibrous material for cleaning having a hydrophilic acrylate or methacrylate coating containing

among others, a bactericide.

  4 - Although the prior art discloses that compositions containing iodine have a broad-spectrum virucidal effect, there has not been, so far, commercially developed a cheap product that can successfully interrupt the spread of viruses such as rhinovirus or influenzavirus iodine causes problems, for example, because of its toxicity and the fact that it is an irritant for animal tissue The action of iodine is non-selective between the

  bacterial and mammalian proteins, and its

  skin may cause severe irritation. In addition, its activity may be reduced or neutralized by

  the action of biological fluids such as blood serum.

  Efforts to modify iodine to avoid

  these difficulties have not, so far, been completely successful.

  There are references in the literature that deal with the bactericidal action of acids such as citric acid, for example, Reid; James D; However, the virucidal action is fundamentally different from the bactericidal action, which is explained by the fact that the viruses and viruses are bacteria are different microorganisms that do not have the same characteristics This is how viruses do not reproduce outside host cells while bacteria do so Quaternary ammonium compounds such as benzalkonium chloride are often effective against bacteria

  but not against viruses such as the various rhinoviruses.

  Although it is known that rhinoviruses are labile in aqueous acid solutions under low pH conditions, eg Davis B D et al; "Microbiology" p 1303 Harper E; Row (Publishers) New York 1973 and Rueckert, R R "Picornaviral Architecture" Comparative Virology Academic Press, New York (1971), p 194-306, the known references do not mention the use of Ce

  concept in epidemiological contexts such as interrup-

  the chain of infection caused by rhinoviruses.

  To our knowledge, the only systematic study of the virucidal action of organic acids (citric, malic, etc.) that can be found in the generally available literature, was carried out by Poli, Biondi, Uberti, Ponti, Balsari and CantonilPoli , G et al: "Virucidal Activity of Organic Acids" Food Chem (England) 4 (4) 251 8 (1979) These researchers found that citric, malic, pyruvic and succinic acids among others, were effective against Herpes virus, orthomyxovirus and rhabdovirus (rabbit virus) Their tests were performed at room temperature

  with aqueous solutions of pure acids No substrates ousup-

  port was used The three viruses chosen to be studied

  by these researchers were all "enveloped" viruses similar to

  In this respect, parainfluenza 3 Poli et al observed that these acids were not effective against adenoviruses which, as is recalled, is a "naked" virus.

  this finding, they conclude that these acids were effective

  caces against viruses "wrapped" but not against viruses

"Naked."

  It is known to those skilled in the art that

  adenoviruses are acid resistant.

  The present invention provides a virucidal product, composition and method which are highly effective over a broad spectrum of viruses and which can be safely produced and used. It results from the recognition that carboxylic acids such as citric, malic and succinic acids when extended in a suitable physiologically acceptable carrier, in addition to their action on certain "enveloped" respiratory viruses, are also effective against rhinoviruses, "naked" viruses. In addition, these acids in the presence of a surfactant Such active agents as sodium dodecyl sulphate are also effective against adenoviruses. In addition, the products according to the present invention may comprise a substrate such as a tissue.

6 2509577

  a non-woven material incorporating such composi-

  According to the process according to the invention, an effective amount of such a composition is brought into contact with the contaminated zone using these products. In general, these compositions can be handled without difficulty and have no harmful effects when In addition, when said compositions are applied to a substrate such as a facial tissue, they have no or very little detrimental effect on the color, odor, resistance or Other important properties The products according to the invention can be used under

  the shape of a dry or wet fabric for example.

  Although the invention will be described in

  referring to preferred embodiments, it is natural-

  Of course, it is intended to cover all the alternatives, modifications and equivalents which fall within the scope of the invention as defined in this invention.

the appended claims -

  The present invention results from an unexpected discovery that certain acids such as citric, malic, succinic and benzoic acids, used in suitable proportions and as will be described hereinafter, have

  high efficacy against rhinoviruses 16, 1 A and 86.

  When used in the presence of a surfactant such as sodium dodecyl sulphate (SDS), these acids are also effective against parainfluenza 3 and adenovirus (the viruses chosen for the study, that is, to say RV-16, RV -IA,

  RV-86, Para-3 and adeno-5, are representative of their class).

  In general, the water-soluble carboxylic acids suitable for carrying out the invention have the following structure

R COOH

  wherein R may be lower alkyls (one to six carbon atoms), lower alkyls substituted eg lower hydroxyalkyl (eg, HOCH 2) 1, lower carboxyalkyl (e.g. E HOOC-CH 2 -CH 2), carboxy, lower hydroxyalkyl (eg HOOCCH 2 CHOH); carboxy halo lower alkyls (for example HOOCCH 2 CH Br); lower carboxy dihydroxyalkyl (eg HOOCCHOH-CHOH-); dicarboxy, hydroxy lower alkyl OH

1

  (e.g. HOOC -CH 2 C CH) COOH lower alkenyls, lower carboxy alkenyls (e.g. HOOC CH = CH -), lower alkenyl dicarboxy (e.g.

LHOOC-CH 2-C = CH -)

  phenyls (e.g., C 6 H 5), substituted phenyls (eg hydroxy phenyl HO-C 6 H 4). Other examples of acids are hydroxy, lower alkyl lactic; carboxy, hydroxy lower alkyl, 2-methyl malic; carboxy, halo, lower alkyl 2-chloro-3-methyl succinic; carboxy dihydroxy lower alkyl, 2-methyl tartaric; dicarboxy, hydroxy lower alkyl, 2-methyl citric; and carboxy lower alkenyl fumaric The above definitions have been given for information only, but not limitatively. The term "substitud" indicates that one or more hydrogen atoms are substituted by halogen atoms (F, Cl, Br, I ), hydroxy groups, amino groups, thiol groups, nitro groups, cyano groups, etc. The surfactant may be nonionic (for example polyoxyethylenated alkylphenols such as TRITON X 100 (registered trademark), manufactured by Rohm and Haas, polyoxyethylenated sorbitol esters such as TWEEN 40 (trademark), manufactured by ICI, USA, Inc) cationic (eg cetylpyridinium chloride)

-8- 2509577

  8 - (6 H 5 N (CH 2) 15 CH 3 Cl-), methylbenzethonium chloride Me 1 + (Me 3 CCH 2 C (Me) 2 -CH (Me) Me) -CH CH 2 CH 2 CH 2 N-CH 2 C 6 H 5 Cl-)

1

  Me or anionic (eg sodium dodecyl sulfate (CH 3 (CH 2) 10-CH 2 SO 3 -Na), 1,4 bis (2-ethylhexyl) ester of the sodium salt of sulfosuccinic acid, such as The preferred anionic surfactants can be represented by the formulas: 1 (RO 503) x or (R 503) x wherein M + is a mono, di or trivalent metal cation or an ammonium ion or a substituted ammonium ion, x is an integer and R is an alkyl group

C H CO R

(12 2 1

2 M-03 S CH C 02 R 2

22

  in which M and x are defined as indicated above and R 1 and R 2 may be the same or different and may be represented by aliphatic groups with straight or branched chains. The anionic surfactants mentioned above are given for illustrative purposes and not In general, surfactants alone have no virucidal effect with respect to

  naked viruses such as rhinoviruses.

  Although the invention is not limited to the use of cellulosic tissue (such as facial tissue, bath towel, hand towels and the like) as a substrate or carrier of virucidal agents, a facial tissue impregnated with these new virucidal agents sufficiently illustrate the

  basic principle of the invention and represents a method of

  This is why, the experiments described in the following paragraphs, have

  performed using facial tissue as a substrate.

  Examples of nonwoven substrates which are suitable for carrying out the invention are wet wiping materials such as wet creped hand towels and non-woven and blown molten polymer-based fabrics commonly used in the manufacture of articles. for hospitals such as surgical linens, sheets, gowns, pillowcases and the like Textile materials of any type with laminates of different materials may be used as a substrate For example, hygienic protective masks used by persons suffering from diseases respiratory

  constitute an excellent means of implementing the invention.

  Other physiologically acceptable, substantially inert carriers, i.e. those which are essentially non-toxic and non-irritating to human or animal tissue under normal conditions of use will readily come to the attention of those skilled in the art. the field of applications such as lotions, sprays, creams, encaustic products

and the like.

  Generally speaking, the prepa-

  Experimental ration of the samples in the examples below was simple and fast KLEENEX facial tissues (registered trademark) with three plies (28 cm x 31 cm, - basis weight about 43 grams per square meter for the three plies together) were impregnated with aqueous acid solutions

  citric, malic, succinic and benzoic by simple soaking.

  The acids were used either alone or in homogeneous mixtures Generally, the impregnating solution contained a small percentage of a surfactant such as Aerosol-OT salt.

  of the 1,4-bis (2-ethylhexyl) ester of the sulfosulfuric acid

  produced by AMERICAN CYANAMID or sodium dodecyl sulfate In some examples, a small amount of glycerol

  was also added to increase the flexibility of the fabric.

  The impregnated fabrics were pressed between rolls to express excess impregnancy and to ensure uniformity of impregnation. The fabrics were weighed, dried and the degree of impregnation (i.e. absorbed) was calculated Tissues were then ready to test for virucidal efficacy.

  The process chosen to test the effectiveness

  The virucidal drug was in accordance with standard virological test (TCID 50) techniques with slight variations required for the presence of the cellulosic substrate.

description of this process follows:

VIRUCID TEST METHOD:

  1 Products used A Solutions 1 Neutralizing solution: 6.4 ml 2 M Na HPO 4 dt 4 1.2 ml 1.0 M Citric acid 92.4 ml 1 x Medium 199 (nutrient medium for tissue culture) 2 Solution of Hank Mc Ilvaine salt (HMSS): 2.0 ml 1.0 Citric acid Diluted to 2 liters with 18.0 ml 2.0 M Na 2 HPO 4 sterile solution of

  The p H of this solution is 7.0 Hank balanced.

  3 Hank salt solution balanced: g / liter in water two distilled laws Na CI 8.0

KCI 0.4

  Mg 504 7 H 20 0.2 Ca CI 2 (anhydrous) 0.14 Na HPO 4 2 H 2 O 0.06 KH 2 PO 4 (anhydrous) 0.06 Glucose 1.0 Red phenol 0.005 Na HCO 3 0.35

  To note: the aforementioned solutions are not virucides.

11 2509577

  B Viruses and cell culture cultures: 1 Rhinovirus type 16, type 1A and type 86

  Rhinovirus types 16, 1 A and 86 (RV 16, 1 A and 86 resp.

  are grown on Hela cell cultures (O-Hela) and stored at 51 ° C until they are used. The virucidal effect test for rhinoviruses is performed in tissue culture test tubes. O-Hela cell incubated on a shaking table

at 33 C.

  2 Parainfluenza type 3: parainfluenza type 3 (Para 3) is cultured on rhesus monkey kidney tissue cultures and stored at 51 ° C until used. The virucidal effect test for para 3 virus is performed using 0-Hela cell tissue culture test tubes incubated in

a stationary position at 33 C.

  Adenovirus type 5: Adenovirus type 5 (Adeno 5) is cultured on cultures of H Ep-2 cell tissue and stored at -51 ° C until the time of use, the test of the virucidal effect interesting the Adeno virus is performed using incubated Human Cell Culture Epithelial Carcinoma -2 (HEP-2) tissue culture test tubes

  in a stationary position at 37 C.

  II Methods A Virucidal effect test A 1: 1 (volume: volume) mixture of virus and saliva was prepared. A 6.5 cm 2 sample was cut from a treated Kimberly-Clark KEENEX tissue and placed on a disk of Petri-plastic (a treated fabric is a fabric

  impregnated with the virucidal agent to be tested).

  saliva (0.1 ml) is pipetted and placed directly on the sample and allowed to react for one minute. It should be noted that the virus dilution is 1/2 after a reaction time of one minute, 5 ml of the The neutralizing solution is pipetted onto the sample placed on the Petri plate and stirred for 3 seconds. The dilution of the virus is then 102. The salivary solution neutralizing solution mixture is then removed from the plate. Pipette and place in a tube containing 5 ml of the Hank-Mc Ilvaine salt solution. The sample is placed in the same tube by tilting the plate and using the end of a pipette to push it to the inside the tube The tube containing the 10 ml of solution and the sample is vortexed for 30 s This tube contains a virus dilution of 10-2.3 or 1: 200 dilution series of 1:10 ( a clean pipette is used for each dilution) t make from the solution of 10-2.3 in

  taking 0.3 ml of this dilution and adding 2.7 ml.

  0.1 ml Hank-Mc Ilvaine salt solution is inoculated into each tissue culture test tube.

  two tubes are inoculated by dilution -

  For each experiment, two sets of checks are performed. The first may be "virus control" as it is intended to control the infectivity of the virus suspension itself without the saliva or tissue substrate. The virus suspension is Serially diluted 1:10 in the HMSS 0.1 ml of specific dilutions is inoculated by test tube containing the cell tissue cultures The information obtained by this control gives the number of infectious virus units that are contained in the virus solution which has been stored at -510 C and ensures that the aliquot of the virus solution used in the experiment has not lost its infectivity during

  freezing, storage or thawing operations.

  The second control, "tissue control" consists of performing the virucidal effect test using 6.5 cm 2 of an untreated KEENEX (trade mark) tissue. The information obtained from this check gives the number of Infectious virus units that can be recovered from 6.5 cm 2 of the untreated tissue after the 13-virucidal effect test The inoculated tissue culture tubes are examined for seven days to detect the viral infection. The end of a virucidal effect test for a given sample is the dilution of the virus that actually produces an infection or is calculated to infect only one of the two inoculated tubes. This number is defined as the infectious dose of tissue culture or TCID 50 The results of

  virucidal activity for a given sample are usually

  The result is that the "log difference" between the TCID 50 result log of the processed sample subtracts from the log

  TCID 50 of the untreated sample.

  The virucidal efficacy of a sample can be calculated from the "log difference" as follows: X -Y virucidal efficacy = x 100% X where: X = initial virus concentration (infectious units /

  0.1 ml) in the untreated sample used as a control.

  Y = the final concentration of the virus (infectious units / 0.1 ml

in the treated sample.

  The following examples explain the method of calculation (in the tests, the final concentration of the virus was always less than or equal to 102 3 infectious units / 0.1 ml). For the majority of the results, the final concentration of the virus was less than 102. , 3 With an initial virus concentration of 0.63, this means a log difference greater than 4 and a higher percentage of "killed" viruses

than 99.99%).

  1 Initial concentration: X = 106.3 Final concentration: y = 1 o 2.3 Log difference = (log 106.3 log 102 ') = 4 -14- Virucidal efficacy = (103 1023) X 100% 106.3

2.3 (104

x 100% 1o 6 3

106 '3

= 99.99%

  2 Initial concentration: X = 104 '8 Final concentration: y = 10 o 2.3 Difference of log = 2.5 Virucidal efficacy = (104' 8 102 '3) x 100% 104.8

= 99.7%

  The calculation method shown above is in

  compliance with standard microbiological testing techniques.

  It provides reliable and reproducible results in

  limits of variability related to biological experiments.

RESULTS

  The results are recorded in the tables

  I, II and III The results given in Table I show

  that simple organic carboxylic acids such as

  citric, malic, tartaric and succinic acids and derivatives

  of these acids (eg 2-bromo-succinic acid).

  nique) and benzoic acid and its substituted derivatives (acid

  salicylic), used on facial tissues in

  are highly virulent against rhinovirus

16 and parainfluenza 3.

  In addition, the results of Table I

  when the acids are used in combination with a surfactant such as Aerosol OT or Dodecyl Sulphate.

  sodium, acid concentrations in the facial tissue may be

  can be lowered without the virucidal effect being altered.

  Table II summarizes the results of the

  with mixtures of acids selected from the group

  killed by citric, benzoic, succinic and malic acids.

  The results show that the facial tissues treated with acid mixtures are virucidal against rhinovirus 16 and parainfluenza 3 They highlight the fact that the facial tissue impregnated with a mixture of acids such as citric acid

  and malic acid and a surfactant such as dodecyl-

  Sodium sulfate (SSD) is effective against rhinoviruses 16, 1A and 86 and adenovirus. As these examples show, simple organic acids such as citric / malic / succinic acids, when used in combination with a suitable surfactant, such as sodium dodecyl sulphate, are

  highly virucidal against the usual respiratory viruses

  among which rhinoviruses 16, 1 A and 86, parainfluenza 3 and adenovirus 5 are typical examples.

  using facial tissues as a means of supporting

  The virucidal substances mentioned are highly effective.

  The interest of the invention lies in the

  it provides the basis for interrupting the chain of infection.

  Because viruses do not reproduce outside the host cell, the degree of inactivation demonstrated in the assays provides a simple and

  practice to reduce the concentration of viruses in the vicinity of a person infected with a respiratory virus This, in turn,

  significantly reduces the tendency of the infection to spread.

TABLE I

  VIRUCID EFFICIENCY OF SIMPLE ACIDS AGAINST

  PARAINFLUENZA 3 VIRUSES (TIME OF EXHIBITION

  Composition virucidea surfactant

RHINOVIRUS 16 AND THE

ONE MINUTE)

  Viricidal efficacy Rhinovirus 16 Parainfluenza 3 citric (23.2%) citric (18.7%) citric (9.7%) citric (9.4%) succinic (20%) succinic (9.1%) 2-bromosuccinic malic (9.4%) (10.4%) tartaric (15%) benzoic (30%) salicylic (18%) salicylic (9%) None None AO Tb (1%), SDC (1%)

SDS (1%)

No

SDS (2%)

SDS (1%)

AOT (0.5%)

  None None None None a The figures in parentheses represent the percentage of chemicals used reported

the weight of the facial tissue.

  b AEROSOL OT, the sodium salt of the 1,4-bis (2-ethylhexyl) ester of sulfosuccinic acid.

C Sodium dodecyl sulphate.

O Ln Or \ O -4 N 4

Example N

  Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid

> 99.7%

> 99.99%

> 99.99%

> 99.99%

> 99.99%

> 99.99%

99.99%

> 99.99%

> 99.99%

> 99.99%

> 99.99%

99.99%

> 99.99%

> 99.99%

> 99.99%

<99.99%

O%

TABLE II

  EFFICACY VIRUCIDE Dg MLANGES OF ACIDS AGAINST RHINOVIRUS 16 AND PARAINFLUENZA 3 VIRUTS (TEM PO S EXPOSURE OF ONE MINUTE) Compositions Acid Citri Qu Benzoic acid, 7, 3, 1 7.1 8.8, 3, 0, 0, 4, 5, 3 O, 2 11, 1, 6 11.1, 6 4.8 13.8, 7 0.2 0.2 0.2 0 $, 2 0.2 0.2 Acidic acids Malic, 2, 3, 2, 1, 6, 3, 6, 3 4.8, 7 9.7 Virucidal Efficacy Acidea Succinic Surfactant, 2, 0 5., 0, 0 A Ob)

AOT (1)

AOT (1)

AOT (1)

AOT (1)

AOT (1)

AOT (1)

AOT (1)

AOT (1)

AOT <1)

AOT (1)

AOT (1)

AOT (0,5)

AOT (1)

AOT (0, 5)

AOT (1)

AOT (1

TX d 100 (2)

SDS (2)

SDS (2)

Rhinovirus 16

> 99.99

*> 99.99

> 99.99

> 99> 99

> 99.99

> 99.99

> 99.99

> 99.99

> 99.99

> 99.99

> 99.99

> 99.99

> 99.99

> 99.99

> 99.99

> 99.99

> 99.99

> 99.99

> 99.97

99.97 Parainfluenza 3

> 99.97

> 99.97

> 99.97

> 99.99

> 99.99

> 99.97

> 99.97

> 99.97

> 99.99

> 99.97

> 99.97

> 99.97

> 99.7

> 99.7

> 99.7

> 99.70

> 99.99

> 99.99

> 99.90

> 99.90

  "Leas figures in parentheses represent the percentage of chemicals used facial tissue b AEROSOL OT

TRITON X-100

  d Dide # sodium cylsulfate based on the weight of J, -.4

Example N O

  -J l A, L41 - -3 'the e - f -13 2 M to 1

TABLE III

  VIRUCID EFFICACY OF MIXTURES OF ACIDS AND SDS AGAINST RHINOVIRUS 16,

  RHINOVIRUS 1A, RHINOVIRUS 86 AND ADENOVIRUS 5 (EXHIBITION TIME OF ONE MINUTE)

Virucidea composition

Acid Acid Agent Tensio-

  Citric Malic active SD Sb Rhinovirus 16 Citric Maligue active SD Sb Rhinovirus 16, 8 11.2 11.4, 8 11.2, 0, 5, 7, 8, 5, 7, 0 2.2 2,3 2,3 2.2 2, 3 2.0

> 99.99

> 99.99

> 99.99

> 99.99

> 99.99

> 99.99

  Efficacy Virucidal Rhinovirus 1 A Rhinovirus 86

> 99.9

> 99.9

  Adenovirus 5 99.90 99.90 99.70 99.99 99.99 99.90 a The figures represent the percentage of products used in relation to the weight of the facial tissue Sodium dodecylsulfate a Gold N o n C -, 4

Example N

  In order to more specifically illustrate the improved effects achieved by the present invention, additional examples have been given in which the concentrations of the selected acidic compositions have been varied and the virucidal activity measured after 1. and 5 minutes These results are shown in Table IV In general, the acidic compositions that fall within the scope of the invention have a high virucidal efficiency, for example in the case of rhinoviruses or parainfluenza, they produce a fall in the log of 2 or a

  greater inactivation in at least one minute For adeno-

  the time will be at least 5 minutes In general, the degree of inactivation is greater after 5 minutes than at the end of

  of a minute, as one might expect.

  minor discrepancies appear in the reported results due to the margin of error and the nature of the test

  art knows that efficiency is also influenced by the amount

  Composition of the composition which is in contact with the virus and which, in turn, depends on the nature of the support. As shown in Table IV below, a relatively thick support with large voids such that the wool can be ineffective. unless treated with large amounts of composition Conversely, a relatively closed lightweight structure such as

  fabric or nonwoven material will require less composition.

  On the basis of the tests described, however, it is possible to

  determine the effectiveness of a given combination of a composi-

  For example, as shown in Table IV, citric acid is effective at high concentrations.

  between 5 and 10% The method used is described below.

below.

  In these examples, the TCID 50 were ob-

  maintained using low pass WI-30 cells from Flow Laboratories Inc. that were initially run at least once to increase their growth potential. The flasks were fractionated in a 1: 2 ratio and seeded onto culture plates. 96-well agglomerated tissue with a flat bottom providing a growth area of 0.32 cm 2 provided by MA Bioproducts Cells were incubated at 370 ° C in 5% CO 2, and after 24 hours were harvested at 80-90% so

  habitual and had a normal appearance before being used

  The medium (2% MM) used for both dilutions and cell maintenance was MEM Eagles with BSS Earles (with glutamine, gentamycin and 2% fetal calf serum). ) Rhinovirus l A was from the National

  Institute of Allergy and Infectious Diseases, Bethesda, Maryland.

  WI-38 cells were allowed to grow in a vial, and

  coltated after they showed cytopathogenic effect (CPE) 4 + two days after inoculation The virus was harvested, divided into aliquots and stored at -70 ° C, and later

  titrated to WI-38 cells on 96-well plates.

  For the test, the medium was removed from

  plates by placing sterile gauze between the plate and the cover

  The six wells used received 0.1 ml of 2% M 4 in wells which were intended to be used as control cells, an additional 0.1 ml of 2% MM was added. In wells intended for

  To receive the compounds, 0.1 ml of the appropriate dilution was added.

  in each of the six wells The virus stock was mixed in equal proportions with 2% MM for the initial dilution One hundredth of a microml of this dilution of virus was then put

  on a disk processed on a Petri disk The virus has been applied

  uniformly on a disc of tissue using a syringe

  microliter The virus was left on the disk for 1 minute or 5 minutes and then 5 ml of 2% MM was put on disk

  of tissue placed on the petri dish and was slightly agitated.

  The disc and the solution were removed, placed in a tube

  sterile and vortexed for 30 seconds.

  counts, representing the first dilution Three dilutions of 1/10 were made from the original tube and 0.1 ml of the four

  dilutions was added to the WI-38 cells in mono-

  Six wells were used for each dilution.

  Untreated tissue controls were performed at 1 and 5 minutes with and without virus and a virus titration was performed.

  The plates were reintroduced for each test.

  cubed at 370 C in CO 2 at 5% for the duration of the test.

  Acids such as sulfamic acids

  and phosphoric have also been shown to be virucidal Cepen-

  However, these acids were found to be damaging to

ports such as fabrics.

TABLE IV

  Glyconic acid and Salicylic VI t, Succinic t i 't, M 4 alic i fi i

2-Bromo-

Succinct if Tartr iqu e he there t '

concen-

tration

% Add.

Tenajo-

active

SDS -% Add.

  2,4 7.2, 4 3.6 1.4 9.2 6.9 4.6 1.8, 5 7.9, 2 2.1 2 e O, 2 11.7 8.8, 9 2 , 3 One minute TCID 50 (Log, 0) Log Loss Tissue Control Contr Treated, Virus

<2, O

<2, O

<2.10

<2, O

<2.0 O

<5.17

<5.25

<2, O

<2, O

<2, O

3.9

<2, O

N / A

<2, O

2.38 2.0 3.33 2.5

<2, O

<2.0 <2.0

<2, O

> 3.25

> 3.25

> 3.25

> 2 nd 33

> 2.33

o o

> 3.25

> 3,325

> 3, O

0.8

> 3.25

N / A

> 3 25

2, .87 1.92 2.5

> 3.25

> 3.25

> 3.25

> 3.25

  Five minutes Log 10 (TCID 50) Z Fall Log Tissue Contr 6 Virus Treaty <2 <2 <2 <2 <2 4.67

<: 5, 4

<2 <2 <2 3.9

<2, O

<2.0 <2.0

<2, O

<2, O

2.5

<2, O

<2.0 <2.0

<2, O

> 2.75

> 2,075

> 2.75

> 2.75

> 2.75

0.5 O

> 2.75

> 2.75

> 3.17

0.4

> 2.75

> 2.75

> 2.75

> 2.75

> 2.75

2.67

> 2.75

> 2.75

> 2.75

> 2.75

  VULCULAR EFFICIENCY (Z KILLED) A FOUR MINUTE MINUTES

> 99.94

> 99.94

> 99.94

> 99.5

> 99.5

o O

> 99.94

> 99.94

> 99, 9

> 99.94

N / A

> 99.94

99.9 98.8 99.7

> 99.94

> 99.94

> 99.94

> 99.94

> 99.82

> 99.82

> 99.82

> 99.8

> 99.8

o

> 99.82

> 99.82

> 99.93

> 99.82

> 99.82

> 99.82

> 99.82

> 99.8

99.8

> 99.82

> 99.82

> 99.82

> 99.82

r N>) c 't,

Example N O

5 TABLE IV (Continued)

Conc

tration

Acid% Adcd.

Tens io-

act if

SDS -Z Add.

  One minute TCID 50 (Log 10) 15 Fall Log Tissue Contr 8 Treated Virus Five Minutes Log 10 (TCID 50) '1.1 Fall Log Controlled Tissue; Viralide Effectiveness Virus (% Inactivation of rhinovirus A) A Five Minute Maleic and Acotinic Minutes of Phosphoric Citric II, I, Citric / Malic i Substrate Wool Base Weight = 27 nrc / sec 2 Polypropylene Protection Mask, Molten soufflé 6.8 4.5 1.8 9.0 6.8 i, 8, 0 7.5, 0 2.0, 0 3.8 2.5 1.0

, 0 / 5.0

  i <2.0 <2.0 2.25 <2.0 <2.0 3.40 <2.0

<: 2, O

<2 g, 0 3.75 <2.0

<2, O

<: 2, O

  4.25 <2.0 <2.0 0.093 rrg / cn 2 0.093 zng / Cr 2

> 3.25

> 3.25

> 3, 00

> 3.25

3.25 i, 85

> 3.25

> 3.25

> 3.25

1.0

> 3, O

> 3, O

*> 3, O

0.75

> 1.75

> 3, O

0.4 4.6 a a a

<: 2, O

  <2.0 <2.0 <2.0 3, 50 <2.0 <2.0 <2.0 <2.0

<2, O

<2.0

<2, O

4.40 <3.0 <2.0 a

> .2,75

> 2.75

> 2.40

> 2.75

1.25

> 2.75

> 2.40

> 2.75

> 2.4

> 3.17

> 3.17

> 3.17

0.77

> 1.40

> 3.17

NA NA <3.0 1.6

> 99.94

> 99.94

> 99.9

> 99.94

98.6

> 99.94

> 99.94

> 99.94

> 99.9

> 99.9

> 99.9

at

> 99.8

> 99.8

> 99.6

> 99.8

> 99.8 (2)

> 99.6

> 99.8 (2)

> 99.6

> 99.93

> 99.93

> 99.93

> 83

> 98.2> 96

> 99.9> 99.93

f "O NA

NA> 97.5

  NA Means not applicable to any of these, particularly with the addition of a surfactant, cytopathic effects may result in good results. Such effects are described by Lennette et al., Diagonist Procedure for Viral, Rickettsial $,% O

  and Chl amydial Infections, 1979, 5 th Ed, p 67 1-

-at

Example N O

1, bw

TABLE V

  Sulfamic acid Mole / cn 2 Tensio-active Acid Addit Z Addit%

2,4 5

7.2 15

2,4 2,4 2,4

SDS 2%

  Virucidal activity (Inactivation of Rhinovirus 16) One minute Five minutes

99> 99.997

99.997> 99.997.

99 99

99> 99.99

Example N O

"I N'o -J -25-

  This is because such acids are soluble

  in water they can be applied to many

  substrates from an aqueous solution with great ease.

  by impregnation, coating or by other means

  such as spraying or photogra-

  When incorporated into substrates, the composition is applied in an amount sufficient to produce a

  Virucidal activity as defined If the limit

  the acid concentration for the product to be effective has not been determined with precision, generally for

  a substrate such as a facial tissue having a base weight of

  taken between 38 and 52 g per square meter (3 folds), the weight of acids such as citric acid, absorbed relative to the weight of

  dry substrate is at least about 2% and preferably

  about 5% Other substrates such as nonwovens

  can be used in the same way.

  With regard to these other substrates,

  prefers, in general, those with high wetting power.

  As we have shown, the low virucidal effect that we obtain

  with substrates such as wool seems to be due to the incapacity

  cited to be able to penetrate such substrates with a virucidal composition. When acid mixtures are used, these can be mixed in varying proportions, but preferably the mixtures contain at least about 0.2 to 10% of each acid based on the weight of the substrate.

after drying.

  In the case where surfactants are incorporated, they are preferentially selected from the group of anionic surfactants and incorporated in a proportion of about 0.05 to 5% by weight of the substrate.

strat after drying.

  In the case where the organic acids having a virucidal activity according to the present invention are applied to other substrates or supports such as lotions, mouthwashes, creams, dispersions, encaustic products and -26-

  similar products, the quantity of acids for which the products

  virucidal properties occur, can be determined by applying

  however, for example, a log fall of 2 or more would mean that 99% or more of the host viruses are inactivated when they come into contact with the compositions.

  antiviral acids according to the present invention.

  Thus, it is clear that

  According to the present invention, a virucidal product is provided which, under normal conditions of use, fully satisfies the objectives and has the advantages as set forth in the present invention.

previous paragraphs.

  Although the invention has been described with reference to specific embodiments, it is obvious

  many alternatives, variants and modifications

  will be known to those skilled in the art in the light of the description

  precedes also the invention aims to cover all the

  alternatives, variations and modifications that fall within

of the appended claims.

27 -

Claims (27)

  1) A method for interrupting or preventing the spread of respiratory viruses comprising   contact of an area containing the virus with a sufficient quantity   to obtain effective virucidal activity from a   having a virucidal activity containing one or more   virucides essentially non-toxic and non-irritating to human or animal tissue, characterized in that each of said acids has the following structure: R COOH   wherein R is selected from the group consisting of lower alkyls; substituted lower alkyls; carboxy lower alkyls; carboxy lower hydroxyalkyls; carboxy halo alkyl E   lower; carboxy dihydroxy lower alkyls; dicarboxy, hy-   lower alkoxy lower hydroxyalkyls; carboxy alkenyls   férieurs; phenyls and substituted phenyl groups.   2) Process according to claim 1,   characterized in that said compositions also contain   a surfactant selected from the group consisting of   nonionic, cationic and anionic surfactants.   3) Method according to one of the claims   2, characterized in that said acid is selected from the group   consisting of citric, malic, succinic, benzol-   qae and their substituted derivatives and mixtures of two or more of these acids.   4) Process according to claim 2, characterized in that said surfactant is selected from the group consisting of polyoxyethylenated alkyl phenols,   polyoxyethylenes sorbitol esters, quaternium ammonium salts   or the ester salts of sulfuric acid and the ester salts of sulphosuccinic acid having the formula: CHCO 2 R 1 (RO), o (O) M or O H 2 (I. M + ('   <RSO) M +, or (ROSO O Mx or -O OS-CHCO 2 R 3 3 'x 3 22 28 - wherein M is a mono, di or trivalent cation or an ammonium ion or a substituted ammonium ion, x is a integer and R is an alkyl group, R 1 and R 2 are straight chains or   branched aliphatic groups identical or different.   5) Process according to claim 4, characterized in that said surfactant is selected from the group consisting of the sodium salt of the ester 1, 4-bis (2-ethylhexyl) sulfosuccinic acid and dodecylsulfate sodium. characterized in that that and malic. characterized in that that and benzoic. 6) -   that said) - that said characterized in that that and succinic. characterized in that that and benzoic. 8) -   said) - said) - characterized in that said that and succinic. Process according to claim 3,   acid is a mixture of Process according to claim 3,   acid is a mixture of citric acids Process according to claim 3,   acid is a mixture of citric acids Process according to claim 3,   acid is a mixture of malic acids Process according to claim 3,   acid is a mixture of malic acids   11) Method according to claim 3,   characterized in thatsaid acid is a mixture of succinic acids nique and benzolque.   12) Virucidal composition comprising a   physiologically acceptable carrier containing a sufficient amount   to achieve effective virucidal activity of one or more   non-toxic and non-irritating acids for the human or animal tissue in the proportion used, characterized in that each of said acids has the following structure: R-COOH   wherein R is selected from the group consisting of lower alkyls; substituted lower alkyls; carbocycle Ky 29 lower alkyls; carboxy hydroxy lower alkyls; carboxy halo lower alkyls; carboxy dihydroxy lower alkyls; carboxy hydroxy lower alkyls; lower alkenyls; carboxy lower alkenyls; dicarboxy lower alkenyls; phenyl and substituted phenyl groups. 13) Composition according to claim 12, characterized in that said composition also contains a surfactant selected from the group consisting of   nonionic, cationic and anionic surfactants.   14) Composition according to the claim   12, characterized in that said acid is selected from the group   composed of citric, malic, succinic,   and their substituted derivatives and mixtures of these acids.    ) Composition according to the claims   12 and 13, characterized in that said surfactant is   selected from the group consisting of polyoxyethylene phenol   thylenated esters, polyoxyethylene esters, quaternary ammonium salts, sulfuric acid ester salts, sulphonic acid salts, and salts of the sulphosuccinic acid ester having the formulas: CH 2 CO 2 R 1)   (i) (R 503>) M, or (ROSO 3) x M + or M (o -00 S-CHCO 2 R 2) in which M is a mono-, di- or trivalent cation or an ammonium ion, or an ammonium ion substituted, x is an integer and   R is an alkyl group, R 1 and R 2 are straight chains or   mified with identical or different aliphatic groups.   16) Composition according to the claim, characterized in that the surfactant is chosen from   the group consisting of the sodium salt of the 1,4-bis (2-   ethylhexyl) sulfosuccinic acid and sodium dodecyl sulfate. 17) Composition according to claim 14, characterized in that said acid is a mixture of acids citric and malic.   18) Composition according to Claim 14   characterized in that said acid is a mixture of citric acid and that and benzolque.   19) 14 characterized in that citric and succinic. ) 14 caracterized in that malic and benzoic. 21) characterized in that malic and succinic. 22) 14 characterized in that succinic and benzoic. 23)   The composition according to Claim 1, wherein said acid is a mixture of acids. The composition according to Claim 1, wherein said acid is a mixture of acids. acids Virucidal product comprising a sub-   an amount sufficient to obtain a virucidal action of a composition containing one or more non-toxic and non-irritating acids for the human or animal tissue in the proportion used, characterized in that each of said acids has the following structure: R-COOH   wherein R is selected from the group consisting of lower alkyls; substituted lower alkyls; carboxy lower alkyls; carboxy hydroxy lower alkyls; carboxy halo alk Yles lower; carboxy dihydroxy lower alkyls; lower alkyl hydroxy dicarboxy lower alkenyls; carboxy lower alkenyls; dicarboxy lower alkenyls; phenyls and groups substituted phenyl.   24) Virucidal product according to the claim   23, characterized in that said composition also contains   a surfactant selected from the group consisting of   nonionic, cationic and anionic surfactants.    ) Virucidal product according to the claims   23 and 24, characterized in that the substrate is selected from the group consisting of a cellulosic tissue, structures   nonwovens and textile materials.   26) Virucidal product according to the claim   characterized in that said acid is selected from the group consisting of citric, succinic, benzoic acids and their substituted derivatives and mixtures of any of these acids, said acid being present in a proportion   about 2% or more based on the weight of the substrate.   27) Virucidal product according to the claim   characterized in that the substrate is a facial tissue cellulose.   28) Virucidal product according to the claim   characterized in that said surfactant is selected from the group consisting of polyoxyethylenated 2-phenylphenols, polyoxyethylenated sorbitol esters, quaternary ammonium salts, sulfuric acid ester salts, salts thereof, and   alkyl sulphonic acid and the salts of the sulphosuccinate having the formulas: 1 90502 R 2 (RSO 3) M, or (ROSO 3) M, or M -03 S-CHCO 2 R 2   in which M + is a mono, di or trivalent cation or an ammonium ion or a substituted ammonium ion, x is an integer and   R is an alkyl group, R 1 and R 2 are straight or branched   identical or different aliphatic groups.   29) Virucidal product according to the claim   characterized in that the surfactant is selected from the group consisting of the sodium salt of the 1,4-bis (2-ethylhexyl) ester of sulfosuccinic acid and the dodecyl sulphate sodium.    ) Virucidal product according to the claim   characterized in that said acid is a mixture 3 citric and malic acids. -32 -   31) Virucidal product according to the claim   characterized in that said acid is a mixture of acids citric and benzoic.   32) Virucidal product according to the claim   characterized in that said acid is a mixture of acids citric and succinic.   33) virucidal product according to claim characterized in that said acid is a mixture of acids malic and benzoic.   34) Virucidal product according to the claim   characterized in that said acid is a mixture of maleic acids lique and succinic.    Virucidal product according to the claim   characterized in that said acid is a mixture of sucrose acids cinic and benzoic.