Antiviral composition comprising chitosan and xanthohumolone

Abstract

The present invention relates to a composition comprising at least chitosan and xanthohumolone, to a process for preparing same and to the uses thereof. More particularly, the invention relates to a composition comprising at least chitosan and xanthohumolone for use as an antiviral composition, in particular in the treatment or prevention of COVID-19.

Description

[0001] ANTIVIRAL COMPOSITION COMPRISING CHITOSAN AND XANTHOHUMO-

[0002] LONE

[0003] Technical field

[0004] The present invention relates to the technical field of antiviral compositions. The invention relates in particular to a composition comprising at least chitosan and xanthohumolone, its preparation process and its uses.

[0005] State of the art

[0006] December 2019 marked the beginning of the COVID-19 pandemic, which has since spread worldwide with more than 350 million cases and over 5.5 million deaths.

[0007] With the virus spreading rapidly, many scientific teams around the world have been working to find solutions to limit the spread of the virus.

[0008] According to the World Health Organization (WHO), the virus can spread when small liquid particles are expelled from the mouth or nose when an infected person coughs, sneezes, talks, or breathes deeply. These particles vary in size, from large "respiratory droplets" to smaller "aerosols."

[0009] The Covid-19 virus, like many airborne viruses, is transmitted primarily through respiratory droplets. These droplets, laden with viral particles, could infect a susceptible individual either through direct contact with a mucous membrane (direct transmission) or through contact with an infected surface via the nasal, oral, or conjunctival mucous membranes (indirect transmission). They can be projected several meters away but do not persist in the air.

[0010] Once inhaled, to infect its host, the COVID-19 virus attaches to a protein present on the surface of cells, particularly lung cells: the ACE2 receptor. This attachment then allows it to penetrate the cell and infect its host.

[0011] One of the strategies being considered aims to prevent the virus from entering the cell by acting on the mechanisms necessary for the virus to attach to its receptor.

[0012] In addition to isolation, ventilation and barrier gestures, the most widespread solution is wearing a mask during close contact to avoid the transmission of droplets and aerosols.

[0013] There are three main types of masks on the market to combat the transmission of airborne pathogens.

[0014] FFP (Filtering Face Piece) type masks are classified according to their capacity to filter aerosols and meet the NF EN 149 standard.

[0015] -FFP1 (80% filtration of aerosols)

[0016] -FFP2 (94% filtration of aerosols) -FFP3 (99% filtration of aerosols).

[0017] This type of mask protects the wearer from inhaling airborne particles and especially larger droplets that could contain infectious agents.

[0018] There are also surgical masks that comply with the NF EN 14683 standard. By preventing the projection of droplets emitted by the wearer of the mask, this type of mask limits contamination of the external environment and other people. There are also several types of surgical masks classified according to their robustness.

[0019] And finally, there are so-called "general public" masks, which are textile masks with guaranteed filtration, and are mostly washable and reusable. They are easily recognizable. They are reserved for use outside the healthcare system.

[0020] “General public” masks have filtration properties greater than 90% of 3 micron particles for category 1 or greater than 70% of these same particles for category 2.

[0021] All masks on the market are effective in limiting the spread of airborne viruses, including SARS-CoV-2, which causes COVID-19, but they also all have the same drawbacks, namely:

[0022] -very limited comfort when breathing, especially during prolonged use,

[0023] -a significant amount of waste due to the short lifespan,

[0024] -masks do not have any specific action to act against viruses, it is only a physical barrier.

[0025] With the emergence of pandemics such as COVID-19, there is a great need to find new solutions to limit the spread of viruses, particularly airborne viruses such as COVID-19.

[0026] One of the solutions proposed by the prior art is the combination of an antiviral vaccine with a means of preventing the spread of viruses.

[0027] However, there are currently no effective solutions to limit the spread of airborne viruses while having a protective antiviral action.

[0028] There is therefore a need for a means of protection aimed at combating the spread and infection of viruses, particularly airborne viruses such as covid-19.

[0029] Summary of the invention

[0030] To meet this need, the invention proposes a new composition comprising a mixture comprising at least chitosan and xanthohumolone.

[0031] Chitosan is a natural biopolymer derived from chitin, particularly from crustaceans.

[0032] Xanthohumolone is a flavonoid derived from hops, particularly from the Humulus Lupulus species. Chitosan and xanthohumolone are molecules that have long been used in many fields such as food, cosmetics, and pharmaceuticals. Their safety profile and innocuousness are already known from the prior art.

[0033] The inventors surprisingly discovered that the combination of chitosan and xanthohumolone made it possible to obtain a composition with a synergistic antiviral effect, particularly against airborne viruses such as covid 19.

[0034] Such a composition is particularly useful in the context of the invention and overcomes the drawbacks of the prior art, it is biocompatible and has significant antiviral activity. Furthermore, the residues resulting from the degradation of the composition according to the invention are not toxic or genotoxic and do not induce inflammation, particularly of the skin or mucous membranes.

[0035] The composition according to the invention has a xantohumolone / chitosan ratio of between 0.2 and 0.7, preferably between 0.4 and 0.6, even more preferably 0.5.

[0036] In one embodiment, the invention relates to a composition comprising a mixture of chitosan and xanthohumolone, as well as a biocompatible solvent such as acetic acid.

[0037] The composition according to the invention is suitable for use in human or animal health, preferably for use as an antiviral or antibacterial composition.

[0038] According to another aspect, the composition according to the invention is also suitable for the treatment of medical devices.

[0039] Advantageously, the composition according to the invention makes it possible to disinfect and protect medical devices to prevent the spread of viruses and / or bacteria.

[0040] Also, the invention also relates to a kit comprising a vaccine and a composition comprising the combination of chitosan and xanthohumolone.

[0041] According to a final aspect, the invention also relates to a process for preparing the composition according to the invention comprising in particular a step of dissolving the chitosan, mixing, heating and adding a xanthohumolone solution.

[0042] Other characteristics and advantages will emerge from the detailed description of the invention and from the examples which are purely illustrative and in no way limitative of the scope of the invention.

[0043] Detailed description of the invention

[0044] Definition

[0045] For the purposes of the invention, “covid-19” means the SARS-CoV-2 coronavirus and all its variants.

[0046] For the purposes of the invention, the term "chitosan" means a copolymer composed of 2-acetamido-2-deoxy-pD-glucose and 2-amino-2-deoxy-|3-D-glucose in varying proportions. Depending on the degree of deacetylation, the molecule is called chitin or chitosan. Chitin, in its natural state, is itself deacetylated to 5 or 15%. It is generally accepted that an amount of amine greater than 7% makes it possible to distinguish chitosan from chitin.

[0047] For the purposes of the invention, the term "microfilm" means a physical protective barrier of a layer of the biopolymer which acts mechanically.

[0048] For the purposes of the invention, a "biocompatible solvent" means a solvent that is compatible with biological tissues. Therefore, the solvent is safe for the body.

[0049] For the purposes of the invention, “cytopathic effect” means alterations.

[0050] Composition according to the invention

[0051] The present invention therefore relates to a composition comprising a mixture comprising at least chitosan and xanthohumolone.

[0052] Advantageously, the composition according to the invention forms a microfilm having significant antiviral activity, in particular against covid 19.

[0053] Advantageously, the combination of chitosan and xanthohumolone has a synergistic antiviral effect.

[0054] According to another advantage, the composition according to the invention is natural, biodegradable and resorbable.

[0055] Preferably, the composition according to the invention also comprises at least one biocompatible solvent. The biocompatible solvent of the composition according to the invention may be chosen from acetic acid, glycolic acid, lactic acid, glutamic acid, glycerol and mixtures thereof. Preferably, the biocompatible solvent is acetic acid.

[0056] Advantageously, the biocompatible solvent according to the invention makes it possible to solubilize the chitosan and xanthohumolone combination to facilitate the method of administration.

[0057] Preferably, the composition according to the invention comprises chitosan with a viscosity of between 35 and 150 cP, preferably a viscosity of between 80 and 100 cP, in particular between 50 and 96 cP, even more preferably between 85 and 95 cP.

[0058] In the context of the invention, the viscosity is preferably measured using a Brookfield device at room temperature. This is a dynamic viscosity measurement method well known to those skilled in the art.

[0059] Advantageously, the viscosity of the chitosan according to the invention makes it possible to obtain a homogeneous composition.

[0060] Preferably, the composition according to the invention comprises chitosan with a molecular weight of between 150 and 500 KDa, preferably between 200 and 300 KDa, even more preferably between 225 and 275 KDa.

[0061] Advantageously, according to the invention, chitosan having a molecular weight of between 150 and 500 KDa has significant antibacterial and antiviral activity. Preferably, the composition according to the invention comprises chitosan having a degree of deacetylation of between 70 and 90%, preferably between 85 and 90%.

[0062] According to a particular embodiment of the invention, the composition according to the invention, with or without biocompatible solvent, also comprises at least one additive.

[0063] The additive according to the invention is preferably chosen from perfume, essential oils, anionic substances.

[0064] When the composition according to the invention comprises an additive such as an anionic substance, the additive may be chosen from Na(2)SO(4) and Na(2)HPO(4).

[0065] According to a particularly preferred embodiment, the composition according to the invention comprises:

[0066] - Between 0.0001 and 0.025% by weight of xanthohumolone

[0067] - Between 0.1 and 10% by weight of chitosan

[0068] - Optionally between 0.1 and 5% by weight of additives

[0069] Preferably, the composition according to the invention has a xantohumolone / chitosan ratio of between 0.2 and 0.7, preferably between 0.4 and 0.6, even more preferably 0.5.

[0070] Advantageously, such a ratio makes it possible to obtain improved antiviral and antibacterial activity.

[0071] The composition according to the invention may be in the form of a gel, solution, cream or spray, preferably the composition is in the form of a spray, in particular a nasal spray.

[0072] Advantageously, when the composition is in the form of a nasal spray, it has significant antiviral efficacy in a single application. In addition, the microfilm formed by the composition according to the invention is very easily resorbable in the nasal mucous membranes. Once applied, the composition lines the nasal mucous membranes and thus prevents the attachment of viruses or bacteria to the cells by physical and chemical protection via the production of an antiviral and antibacterial microfilm.

[0073] According to another aspect, the invention relates to a medical device comprising the composition according to the invention. Preferably, the medical device according to the invention may be a mask such as a surgical mask, an FFP type mask or a fabric mask.

[0074] Process for preparing the composition according to the invention

[0075] According to another aspect, the composition according to the invention comprises at least the implementation of the following steps: a) Dissolution of the chitosan b) Mixing with stirring of the dissolved chitosan from step a) c) Adjustment of the pH of the mixture from step b) d) Heating of the mixture from step c) e) Addition of a xanthohumolone solution to the mixture from step d)

[0076] Preferably, the dissolution of the chitosan is carried out in a biocompatible solvent at a concentration of between 0.2 and 2% at a ratio of 0.1:1, 1:1 or 2:1.

[0077] The biocompatible solvent of step a) may be chosen from acetic acid, glycolic acid, lactic acid, glutamic acid, glycerol and mixtures thereof.

[0078] Preferably, the biocompatible solvent of step a) is acetic acid.

[0079] Preferably, the mixing of step b) is carried out in a paddle mixer at a rotation speed of between 40 and 150 rpm (rpm = revolutions per minute), preferably between 50 and 100 rpm.

[0080] The mixing in step b) is preferably carried out at room temperature.

[0081] According to a preferred embodiment, the mixing of step b) is carried out for 4 to 12 hours, preferably for 6 to 8 hours.

[0082] The pH adjustment of step c) is carried out by adding a base to the mixture of step b) until a pH between 5 and 7 is obtained, preferably 5.5 to 6.5.

[0083] The heating in step d) is carried out at a temperature between 30 and 100°C, preferably between 80 and 90°C.

[0084] Preferably, the heating of step d) is carried out in a water bath.

[0085] According to a preferred embodiment, the heating of step d) is carried out for a period of 15 minutes to 1 hour, preferably 30 minutes.

[0086] Preferably, the heating of step d) is carried out with gentle stirring.

[0087] According to a particularly preferred embodiment, the xanthohumolone solution from step e) is added at a concentration between 10 and 100 pg / ml, preferably between 40 and 70 pg / ml.

[0088] Kit:

[0089] According to another aspect, the invention relates to a kit comprising the composition according to the invention and a vaccine.

[0090] The different components of the kit can be used simultaneously or sequentially.

[0091] Preferably, the vaccine is an antiviral vaccine, preferably an anti-covid-19 vaccine.

[0092] According to a particular embodiment, the kit according to the invention comprises a vaccine chosen from: Tozinaméran (Pfizer-BioNTech), mRNA-1273 (Moderna), Gam-COVID-Vac (Sputnik V), AZD1222 (Oxford-AstraZeneca), Ad26.COV2.S (Janssen / Johnson & Johnson), Ad5-nCoV / Convidecia (Can-Sino Biologies), BBIBP-CorV (Sinopharm), WIBP (Sinopharm), CoronaVac (Sinovac Biotech), BBV152 / Covaxin (Bharat Biotech), CoviVac (Chumakov), ZF2001 / RBD-Dimer (Anhui Zhifei Longcom), Epi-VacCorona (Vector). According to a particularly preferred embodiment, the kit according to the invention comprises a vaccine administered nasally, such as FLUENZ TETRA (AstraZeneca).

[0093] Uses

[0094] According to a final aspect, the invention relates to the composition according to the invention for its use as a medicament, preferably as an antiviral composition or antibacterial composition.

[0095] The composition according to the invention can be used as an antiviral or antibacterial drug.

[0096] Thus, another subject of the invention is a composition for its use in the prevention and / or treatment of viral and bacterial diseases, preferably for preventing and / or combating covid-19, herpes simplex 1 and 2, cytomegalovirus, hepatitis A, papillomavirus, human immunodeficiency virus as well as sexually transmitted diseases.

[0097] Examples include the prevention and / or treatment of influenza, caused by the influenza virus, but also the prevention and / or treatment of severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), covid-19 disease, caused by coronaviruses, and the prevention and / or treatment of angina, caused by adenoviruses or staphylococcus and streptococcus bacteria.

[0098] Preferably, the composition according to the invention is an antiviral medicament for use in the treatment or prevention of covid-19.

[0099] Advantageously, the composition according to the invention forms a microfilm on the skin or mucous membranes, thus making it possible to prevent the virus from attaching to the cells and to have antiviral activity against it.

[0100] According to another embodiment, the invention relates to the use of a composition as described in the present application for treating medical devices, preferably masks such as surgical masks, FFP type masks and fabric masks.

[0101] Advantageously, the treatment of masks with the composition according to the invention makes it possible to upgrade masks or visors by increasing their usage time and providing them with antiviral activity, in particular against covid-19.

[0102] According to a particular embodiment, the composition according to the invention can be used as a food supplement.

[0103] Thus, the composition according to the invention can be integrated into another composition, in particular into a nutritional composition presented in a form chosen from tablets, capsules, gelcaps, powders, solutions, microcapsules, suspensions, emulsions, food supplements, drinks, and food for humans or animals.

[0104] Examples

[0105] Example of composition according to the invention 1:

[0106] Composition 1 has the following characteristics:

[0107] - Chitosan (260 KDa and 80 cP) = 1% by weight of the composition

[0108] - Xanthohumolone = 0.025% by weight of the composition

[0109] - Essential oil = 0.1% by weight of the composition.

[0110] The pH of the composition is adjusted to be between 5.1 and 6.

[0111] Composition 2 has the following characteristics:

[0112] - Chitosan (260 KDa and 80 cP) = 1% by weight of the composition

[0113] - Xanthohumolone = 0.025%

[0114] - Acetic acid = 0.5%

[0115] - Essential oil = 2%

[0116] The pH of the composition is adjusted to be between 5.1 and 6.

[0117] Example 3:

[0118] Composition 1 has the following characteristics:

[0119] - Chitosan (150 KDa and 50 cP) = 0.5% by weight of the composition

[0120] - Xanthohumolone = 0.005%

[0121] - Anionic substance (Na(2)SO(4) and Na(2)H PO(4)) = 2 to 6%.

[0122] The pH of the composition is adjusted to be between 5.5 and 6.2.

[0123] Example of a process for preparing a composition according to the invention:

[0124] The preparation process includes the implementation of the following steps:

[0125] - Dissolution of chitosan in a 2% acetic acid solution at a ratio of 1:1

[0126] - Mix at room temperature with stirring at 100 rpm for 7 hours

[0127] - Adjust pH to 5.5 to 6.2 by adding NaOH

[0128] - Heating in a water bath at 80°C for 4 hours

[0129] - Addition of a xanthohumolone solution at 20 to 100pg / ml

[0130] The preparation process includes the implementation of the following steps:

[0131] - Dissolution of chitosan in a 1% glycerol solution at a ratio of 2:1 - Mixture at room temperature with stirring at 80 rpm for 6 hours

[0132] - Adjust pH to 5.5 to 6.2 by adding NaOH

[0133] - Heating in a bain-marie at 45°C for 3 hours

[0134] - Addition of a 20 pg / ml xanthohumolone solution

[0135] Example 6:

[0136] Testing antiviral efficacy on a strain of coronavirus 229E

[0137] The composition according to the invention was formulated in gel form and tested on a strain of coronavirus 229E according to standard NF 14476+A2 dating from July 2019.

[0138] Test method

[0139] A) Virus

[0140] Name: HCOV 229 E

[0141] Product Type: Animal Virus

[0142] Classification: Coronaviridae, Coronavirus

[0143] Strain designation: 229 E

[0144] B) Cells

[0145] Name: VERO

[0146] Product Type: Animal Cell

[0147] Organism: Cercopithecus aethiops

[0148] Morphology: epithelial

[0149] Tissue: Kidney

[0150] Physiological state: normal

[0151] C) Experimental conditions

[0152] Test concentrations: 80% (V / V) of a composition according to the invention comprising 1% chitosan and a Xanthohumolone content of 50 pg / mL

[0153] Contact time: 60 minutes ± 10 seconds

[0154] Interfering substance: 0.3g / L BSA

[0155] Phosphate buffered saline: PBS

[0156] Product diluent: Water PPI

[0157] Culture medium: EMEM

[0158] Molecular Sieve Filter: Sephadex LH20 Appearance of product dilutions: Liquid

[0159] Stability and appearance of the mixture during the test: Homogeneous preparation during the duration of the test

[0160] Titration of viral suspension:

[0161] The titration is carried out in microplates by the suspension cell method according to standard NF EN 14476+A2. The cytopathic effect is determined after 4 days of culture.

[0162] The estimation of the number of infectious units is determined by the SPAERMAN-KARBER method well known to those skilled in the art. The SPAERMAN-KARBER method consists of testing according to a geometric progression, that is to say with a constant ratio between successive dilutions, and inoculating a constant volume of each dilution.

[0163] This method allows the determination of Ig DICT50, i.e. the dilution of the virus solution causing cytopathic effects on 50% of the cells in culture.

[0164] An efficiency check is planned by stopping the reaction in ice for 30 minutes before the virus titration is carried out.

[0165] Direct cytotoxicity test:

[0166] This test makes it possible to highlight the necessary concentration of the composition according to the invention to cause toxicity on the VERO cell line.

[0167] Indirect cytotoxicity test: molecular sieving

[0168] The stopping of the activity of the composition according to the invention is carried out by the molecular sieving technique using a sieve filter, i.e. Sephadex LH 20. The neutralization of the composition according to the invention is evaluated by passing through Sephadex after an intermediate dilution. The neutralization of the product is validated by passing through Sephadex from the dilution to 1 / 10. The neutralization of Formaldehyde is validated by passing through Sephadex from the dilution to 1 / 20.

[0169] Cell sensitivity to viruses:

[0170] Comparative titrations are carried out on cells treated or not with the composition according to the invention at a subcytotoxic concentration in order to verify that this does not modify the behavior of the recipient cells with respect to the virus.

[0171] Results :

[0172] 1) Title of the test viral suspension: Ig DICT50: 7.1

[0173] 2) Viral control of the test: Ig TCID50 = 4.88

[0174] 3) Direct cytotoxicity: non-cytotoxic concentration from 8*10-1% (weight / volume)

[0175] 4) Indirect cytotoxicity: non-cytotoxic concentration from 4*10-1% (weight / volume)

[0176] Cell sensitivity to viruses:

[0177] Condition a: VERO cells in the presence of the composition according to the invention: Ig DICT50 = 5.75 Condition b: VERO cells in the absence of the composition according to the invention: Ig DICT50 = 5.63

[0178] Condition b - Condition a = 0.12 < llg

[0179] The sensitivity is therefore valid.

[0180] Control of the effectiveness of stopping the disinfectant activity at T=0

[0181] Condition a: VERO cells in the presence of the composition according to the invention: Ig DICT50 = 6.25

[0182] Condition b: VERO cells in the absence of the composition according to the invention: Ig DICT50=5.88 Difference < 0.5 therefore valid control.

[0183] Test result:

[0184] [Table 1]

[0185] * 1 to 4: Presence of virus, with cytopathic effect observed in 8 cell wells

[0186] 0: Absence of virus, with absence of cytopathic effect

[0187] Conclusion :

[0188] According to the protocol of standard NF EN 14476 + A2, the composition according to the invention induces a reduction in the viral load of coronavirus 229E of 3.75 Ig or 99.98% after an exposure of 60 minutes.

[0189] Example 7:

[0190] Determination of the virucidal activity of chitosan and xanthohulomone products according to the protocol of standard NF EN 14476+A2 (July 2019) coronavirus 229 E.

[0191] Sample identification:

[0192] - Composition 1: chitosan 0.1%

[0193] - Composition 2: chitosan 1%

[0194] - Composition 3: chitosan 1% + 2 mg / mL hops - Composition 4: chitosan 1% + 50 pg hops

[0195] Product appearance: liquid gel with deposits for compositions 3 and 4.

[0196] - Composition 5: 5 pg / mL of Xanthohumolone

[0197] - Composition 6: 50 pg / mL of Xanthohumolone

[0198] - Composition 7: 250 pg / mL of Xanthohumolone

[0199] Product appearance: Solid to be reconstituted in 60% ethanol solution at 5mg / ml

[0200] Test method

[0201] A) Virus

[0202] Name: HCOV 229 E

[0203] Product Type: Animal Virus

[0204] Classification: Coronaviridae, Coronavirus

[0205] Strain designation: 229 E

[0206] B) Cells

[0207] Name: VERO

[0208] Product Type: Animal Cell

[0209] Organism: Cercopithecus aethiops

[0210] Morphology: epithelial

[0211] Tissue: Kidney

[0212] Physiological state: normal

[0213] B) Experimental conditions

[0214] Test concentrations: 80% (V / V) of a composition according to the invention comprising 1% chitosan and a Xanthohumolone content of 50 pg / mL

[0215] Contact time: 60 minutes ± 10 seconds

[0216] Phosphate buffered saline: PBS

[0217] Product diluent: Water PPI

[0218] Culture medium: EMEM

[0219] Molecular Sieve Filter: Sephadex LH20

[0220] Appearance of product dilutions: Liquid

[0221] Stability and appearance of the mixture during the test: Homogeneous preparation during the duration of the test

[0222] Viral suspension titration: The titration is carried out in microplates using the suspension cell method. The cytopathic effect is determined after 4 days of culture.

[0223] The estimation of the number of infectious units is determined by the SPAERMAN-KARBER method well known to those skilled in the art. The SPAERMAN-KARBER method consists of testing according to a geometric progression, that is to say with a constant ratio between successive dilutions, and inoculating a constant volume of each dilution.

[0224] This method allows the determination of Ig DICT50, i.e. the dilution of the virus solution causing cytopathic effects on 50% of the cells in culture.

[0225] An efficiency check is planned by stopping the reaction in ice for 30 minutes before the virus titration is carried out.

[0226] Direct cytotoxicity test:

[0227] This test makes it possible to highlight the necessary concentration of the composition according to the invention to cause toxicity on the VERO cell line.

[0228] Indirect cytotoxicity test: molecular sieving

[0229] The stopping of the activity of the composition according to the invention is carried out by the molecular sieving technique using a sieve filter, i.e. Sephadex LH 20. The neutralization of the composition according to the invention is evaluated by passing through Sephadex after an intermediate dilution. The neutralization of the product is validated by passing through Sephadex from the dilution to 1 / 10. The neutralization of Formaldehyde is validated by passing through Sephadex from the dilution to 1 / 20.

[0230] Cell sensitivity to viruses:

[0231] Comparative titrations are carried out on cells treated or not with the composition according to the invention at a subcytotoxic concentration in order to verify that this does not modify the behavior of the recipient cells with respect to the virus.

[0232] Results of compositions 1 to 4:

[0233] 1) Direct cytotoxicity: non-cytotoxic concentration from 1*10-3% (volume / volume)

[0234] 2) Indirect cytotoxicity: non-cytotoxic concentration from 1*10-2% (volume / volume)

[0235] Sensitivity of cells to viruses of compositions 1 to 4:

[0236] Condition a: VERO cells in the presence of the composition according to the invention: Ig DICT50 = 7.38

[0237] Condition b: VERO cells in the absence of the composition according to the invention: Ig DICT50=7.13

[0238] Condition b - Condition a = 0.25 < llg

[0239] The sensitivity is therefore valid.

[0240] Control of the effectiveness of stopping the disinfectant activity at T=0

[0241] Condition a: VERO cells in the presence of the composition according to the invention: Ig DICT50 = 6.25 Condition b: VERO cells in the absence of the composition according to the invention: Ig DICT50 = 6.88 Difference < 0.5 Ig therefore valid control.

[0242] Viral witness to the trial:

[0243] - Before Sephadex: Ig DICT50 = 6 - After Sephadex: Ig DICT50 = 5.75

[0244] Difference = 0.25 < 0.5 Ig so the control is valid

[0245] Test results:

[0246] [Table 2]

[0247] * 1 to 4: Presence of virus, with cytopathic effect observed in 8 cell wells 0: Absence of virus, with absence of cytopathic effect.

[0248] [Table 3]

[0249] * 1 to 4: Presence of virus, with cytopathic effect observed in 8 cell wells 0: Absence of virus, with absence of cytopathic effect.

[0250] Results of compositions 5 to 7: 1) Direct cytotoxicity: non-cytotoxic concentration from 2.5*10-1 pg / ml (weight / volume)

[0251] 2) Indirect cytotoxicity: non-cytotoxic concentration from 2.5 pg / ml (weight / volume) Sensitivity of cells to viruses of compositions 5 to 7:

[0252] Condition a: VERO cells in the presence of the composition according to the invention: Ig DICT50 = 7.13

[0253] Condition b: VERO cells in the absence of the composition according to the invention: Ig DICT50=7.13

[0254] Condition b - Condition a = 0 < llg

[0255] The sensitivity is therefore valid.

[0256] Control of the effectiveness of stopping the disinfectant activity at T=0

[0257] Condition a: VERO cells in the presence of the composition according to the invention: Ig DICT50 = 6.75

[0258] Condition b: VERO cells in the absence of the composition according to the invention: Ig DICT50=6.88 Difference < 0.5 Ig therefore valid control.

[0259] Viral witness to the trial:

[0260] - Before Sephadex: Ig DICT50 = 6

[0261] - After Sephadex: Ig DICT50 = 5.75

[0262] Difference = 0.25 < 0.5 Ig so the control is valid

[0263] [Table 4]

[0264] * 1 to 4: Presence of virus, with cytopathic effect observed in 8 cell wells 0: Absence of virus, with absence of cytopathic effect

[0265] Conclusion :

[0266] [Table 5]

[0267] The results of the reduction of the viral load of coronavirus 229 E after a contact time of

[0268] 60 min with the chitosan product alone (composition 1 or 2), the chitosan + hops combination (composition 3 or 4) and xanthohumolone alone (compositions 5 to 7) show a lower cytopathic effect on Coronavirus 229 E than that of the chitosan / xanthohumolone combination according to the invention.

Claims

Demands

1. Composition comprising a mixture including at least chitosan and xanthohumolone.

2. Composition according to claim 1, characterized in that said composition also comprises a biocompatible solvent.

3. Composition according to the preceding claim, characterized in that the biocompatible solvent is selected from acetic acid, lactic acid, glycolic acid, glutamic acid, glycerol or mixtures thereof.

4. Composition according to any one of the preceding claims, characterized in that the xanthohumolone / chitosan ratio is between 0.2 and 0.

7.

5. Composition according to any one of the preceding claims, characterized in that the chitosan has a viscosity between 70 and 150 cP

6. Composition according to any one of the preceding claims, characterized in that the chitosan has a molecular weight between 150 and 500 kDa

7. Composition according to any one of the preceding claims, characterized in that it is in the form of a gel, solution, spray, cream.

8. Composition according to any one of the preceding claims for its use as a medicinal product.

9. Composition according to any one of claims 1 to 7 for its use as an antiviral and / or antibacterial composition.

10. Composition according to any one of claims 1 to 7 for its use in the treatment and / or prevention of viral and / or bacterial diseases.

11. Composition according to any one of claims 1 to 7 for its use in the treatment and / or prevention of COVID-19 and all its variants.

12. Use of a composition according to any one of claims 1 to 7 in the processing of medical devices.

13. Kit comprising the composition according to any one of claims 1 to 7 and a vaccine.

14. Medical device comprising the composition according to any one of claims 1 to 7.

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