Functionalized inorganic oxide or hydroxide for use against enveloped viruses

Abstract

The present invention provides materials based on inorganic oxides, such as silica, and a photosensitizer, such as a porphyrin type macrocycle. In the material of the invention, the photosensitizer is covalently linked to the surface of the inorganic oxide. It has been observed that the materials of the invention exhibit outstanding antiviral properties against enveloped viruses, such as coronaviruses or hepaciviruses. The invention also provides the use of these materials to deactivate enveloped viruses, for example to decontaminate a fluid. The materials of the invention can for example be used in devices such as filters or respiratory masks.

Description

[0001] FUNCTIONALIZED INORGANIC OXIDE OR HYDROXIDE FOR USE AGAINST ENVELOPED VIRUSES

[0002] The present invention relates to materials having anti-viral properties, in particular to materials of oxide type functionalized with a photosensitizer.

[0003] An enveloped virus is a type of virus that possesses a lipid envelope surrounding its nucleocapsid, which contains the genetic material of the virus. The envelope is derived from the plasma membrane of the host cell and contains viral proteins involved in host cell attachment and entry. This envelope plays a vital role in viral infection and transmission, and it is also a target for many antiviral agents. Coronaviruses, Retroviruses, Flaviviruses are examples of enveloped viruses. HCoV-229E is a common Alphacoronavirus that causes mild symptoms of common cold in humans, SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus-2) and MERS-CoV (Middle East Respiratory Syndrome Coronavirus) are two highly pathogenic coronaviruses that belong to the Betacoronavirus genus. Another example of an enveloped virus is Hepatitis C, or HCV, that belongs to the Flaviviridae family and which is the main cause of viral chronic hepatitis worldwide.

[0004] In many cases, there is no efficient treatment for curing diseases caused by enveloped viruses. For example, up to this day, there exists no treatment for the diseases caused by MERS-CoV and treatments against SARS-CoV-2 are not very effective and are reserved for certain populations such as the immunocompromised patients.

[0005] UV irradiation and the use of quaternary ammoniums break down viral envelopes. Filters or masks stop most, but not all, viruses. These are all ways of hindering the spread of enveloped viruses.

[0006] There is therefore a need for other means of deactivating enveloped viruses either for treating diseases caused by enveloped viruses or for disinfecting surfaces or fluids.

[0007] In particular, materials (as opposed to molecules and ions) having a permanent antiviral activity against enveloped viruses could be of interest, for example, for self -cleaning surfaces, for the treatment of contaminated fluids or even for the treatment of diseases. One interest of a material over a molecule is that it is easier to recover.

[0008] In a previous study (Meunier T, et al., A Photoactivable Natural Product with Broad Antiviral Activity against Enveloped Viruses, Including Highly Pathogenic Coronaviruses. Antimicrobial Agents and Chemotherapy, Vol. 66, Issue 2, eO 1581 -21 ), it was shown that Pheophorbide A possesses antiviral properties against enveloped viruses under visible light exposure, with the mechanism of action being the production of singlet oxygen that is capable of oxidizing the lipid envelope of enveloped viruses.

[0009] Pheophorbide has the following formula (I):

[0010] I

[0011] However, in Meunier et al., it was not suggested to include Pheophorbide A in a material. A fortiori, it remained unclear if the observed activity can be maintained if Pheophorbide A is included in a material. A fortiori, it remained unclear which modes of inclusion of Pheophorbide A or any other photosensitizer in a material could provide the best antiviral activity.

[0012] In an effort to solve the above problems the present invention provides the items and embodiments described below.

[0013] 1. Item 1 : Material comprising an outside surface and an inorganic oxide or hydroxide of an element E, said outside surface comprising a photosensitizer P, and said inorganic oxide or hydroxide comprising surface oxygen atoms O1covalently linked to said photosensitizer.

[0014] It has been observed that a material as described above had strong antiviral properties against enveloped viruses, including HCoV-229E, SARS-CoV-2, MERS-CoV and HCV. Moreover, it has been observed that the antiviral efficiency of said material was higher than that of other materials, such as metal-organic frameworks (MOFs), functionalized with the same photosensitizer. Moreover, these materials are stable in water.

[0015] As defined herein, an inorganic oxide of an element E is a material consisting of the element E and oxo ligands and wherein all the atoms of E are surrounded by oxo ligands. The surface of the oxide may comprise other types of ligands.

[0016] As defined herein, an inorganic hydroxide of an element E is a material consisting of the element E and hydroxo ligands and wherein all the atoms of E are surrounded by hydroxo ligands. The surface of the hydroxide may comprise other types of ligands.

[0017] It should be understood that the surface oxygen atoms of the inorganic oxide or hydroxide are the oxygen atoms situated at the surface of the inorganic oxide or hydroxide.

[0018] A photosensitizer is defined herein as a molecule that can produce reactive oxygen species when excited by an appropriate light source in the presence of oxygen.

[0019] 2. Item 2: The material of item 1, wherein the element E is chosen from Si, Al, Ti, Zr, Hf, V, Nb, Ta, Fe, Sb, W, Cu, Ce, Y, and mixture thereof, preferably chosen from Si, Al, Ti, Zr, Hf, and mixtures thereof, more preferably chosen from Si, Al, Ti, and mixtures thereof, most preferably the inorganic oxide or hydroxide is a silicon oxide.

[0020] 3. Item 3: The material of item 1 or item 2, wherein the inorganic oxide or hydroxide is not in the form of nanoparticles.

[0021] 4. Item 4: The material of item 1 or item 2, wherein the inorganic oxide or hydroxide is in the form of nanoparticles.

[0022] The material of the invention can be in various forms that may or may not be nanoparticles.

[0023] As defined herein a nanoparticle is a particle with a hydrodynamic size of 1 micrometer or lower as measured by dynamic light scattering, for example using an instrument such as a Zetasizer Nano ZS (Malvern Instruments, Orsay, France).

[0024] For example, the material of the invention may be in the form of particles with a hydrodynamic size of 1 micrometer or higher as measured by dynamic light scattering, for example using an instrument such as a Zetasizer Nano ZS (Malvern Instruments, Orsay, France).

[0025] The hydrodynamic size (also called Z-average size) is defined by the ISO norm: ISO 22412:2017.

[0026] The material of the invention can be used in various forms, including as a coating, suspended in a solution or dispersed in a matrix.

[0027] 5. Item 5: The material of any one of items 1 to 4, wherein the inorganic oxide or hydroxide is obtained by a sol-gel process comprising the condensation of a precursor of the inorganic oxide or hydroxide.

[0028] As defined herein, a sol-gel process is a process for producing a solid material from precursors dissolved in a liquid medium or suspended therein comprising the condensation of the precursor. For example, a precursor dissolved in a liquid medium (e.g. comprising water) can condense into a colloidal solution (sol) that acts as the precursor for an integrated network (or gel) that may be the material or maybe further processed to obtain the material.

[0029] For some precursors, such as alkoxide precursors in the presence of water, the condensation comprises the hydrolysis of the precursor.

[0030] 6. Item 6: the material of item 5, wherein the condensation occurs in an aqueous medium.

[0031] For example, the reaction medium can comprise at least 20% w / w of water relative to the total weight of the reaction medium. Other solvents can be present. These other solvents may include alcohols such as methanol, ethanol, propanol, butanol and their isomers. When alkoxide precursors are used, the hydrolysis of said precursors during the sol gel process results in the presence of alcohol in the reaction medium even if no alcohol was added to the reaction medium.

[0032] 7. Item 7: the material of item 5 or 6, wherein the sol-gel process comprises the calcination of the product of the condensation.

[0033] For example, calcination can be performed by heating in air at a temperature of above 500°C and for at least 5 hours. Before the calcination, the condensation can include a step of drying, for example by heating the reaction medium in air at a temperature of above 50°C for at least 10 hours. 8. Item 8: the material of any one of items 5 to 7, wherein the sol-gel process is acid catalyzed.

[0034] As defined herein the sol-gel process is acid catalyzed when an acid (a Lewis acid or a Brpnsted acid) is added to the reaction medium to catalyze the condensation of the precursor. The sol-gel process can also be a base catalyzed sol-gel processes or an uncatalyzed sol-gel processes. An uncatalyzed sol-gel process corresponds to the case where the condensation of the precursor occurs without addition of an acid or a base. An example of a basecatalyzed sol-gel processes would be a process wherein the condensation of a silicon alkoxide is performed in the presence of ammonia (in this case, ammonia may be added as an aqueous solution). Preferably, the acid catalyzed sol-gel process comprises the addition of a Brpnsted acid, for example HC1, to the reaction medium. When the condensation comprises the hydrolysis of the precursor, such as when an alkoxide precursor is used, the acid catalysis may comprise the catalysis of the hydrolysis of the precursor by the added acid.

[0035] It has been observed that when the inorganic oxide or hydroxide is obtained via an acid catalyzed sol-gel process, the antiviral properties of the material are enhanced compared to the case where a base catalyzed or a non-catalyzed sol-gel process is used.

[0036] 9. Item 9: The material of any one of items 5 to 8, wherein the precursor is an alkoxide of the element E.

[0037] As defined herein an alkoxide of an element E is a molecule comprising the moiety E-O- Aik, where Aik is an alkyl chain. Preferably, the element E is directly linked to a number of OAlk moieties corresponding to the number of valencies of element E. For example, in this case the precursor may be a tetraalkoxysilane, a trialkoxide of aluminum(III), a tetraalkoxide of titanium(IV), etc.

[0038] 10. Item 10: the material of any one of items 1 to 9, wherein the inorganic oxide or hydroxide is a mesoporous oxide or hydroxide.

[0039] As defined herein, a mesoporous oxide or hydroxide is an oxide or a hydroxide containing pores with an average size of between 2 and 50 nm as measured by the Barrett-Joyner-Halenda (BJH) procedure.

[0040] 11. Item 11: the material of item 10, wherein the inorganic oxide is a SBA-15 mesoporous silica.

[0041] Santa Barbara Amorphous-15 (SBA-15) is a highly stable mesoporous silica. It features a framework of uniform hexagonal pores with a narrow pore-size distribution and a tunable pore diameter of 5nm to 15nm. SBA-15 is synthesized in acidic conditions using triblock copolymer Pluronic 123 (E020P070E020) as a templating agent and tetraethoxysilane (TEOS) for the source of silica. After condensation, the organic template may be removed by a variety of methods, including calcination; reflux extraction; H2O2 treatments; microwave digestion; and / or washing with pure solvents, ethanol, acetone, or water.

[0042] 12. Item 12: The material of any one of items 1 to 11, wherein the photosensitizer is a heterocyclic macrocyclic photosensitizer.

[0043] Examples of heterocyclic macrocyclic photosensitizers include but are not limited to porphyrins, verteporphins, chlorins, bacteriochlorins, isobacteriochlorins, texaphyrins, porphycenes, hemiporphycenes, corrphycenes, phthalocyanines, naphthalocyanines, and sapphyrins. As defined herein, when the heterocyclic macrocyclic photosensitizer is a ligand, it can be metalated or not. 13. Item 13: The material of item 12, wherein the heterocyclic macrocyclic photosensitizer is a pyrrolic macrocycle.

[0044] As defined herein a pyrrolic macrocycle is a macrocycle comprising at least one pyrrole moiety.

[0045] 14. Item 14: The material of item 13, wherein the pyrrolic macrocycle is a porphyrin-type macrocycle.

[0046] As defined herein a porphyrin-type macrocycle is a molecule comprising moiety (II) or the corresponding metalated moiety:

[0047] II wherein C1, C2, C4, C5, C7and C8may be sp2 or sp3 hybridized and wherein any of C'-C12may bear at least one substituent.

[0048] It should be understood that the carbons of the porphyrin-type macrocycle that may bear at least one substituent may bear zero substituent (then the valency of said carbon is completed with one or two hydrogen atoms depending on the hybridization of said carbon). Moreover, if said carbon is sp2 hybridized, it may bear one substituent at most, and if said carbon is sp3 hybridized, it may bear two substituents at most.

[0049] 15. Item 15: The material of item 14, wherein C3, C6, C9, and C12are directly linked to an atom selected from H and a sp3 carbon atom.

[0050] 16. Item 16: The material of item 14 or 15, wherein the porphyrin-type macrocycle comprises a substituent selected from alkyl moieties and aliphatic moieties on a carbon selected from C1, C2, C4, C5, C7, C8, C10, and C11.

[0051] 17. Item 17: The material of item 16, wherein said substituent is selected from methyl, ethyl and vinyl groups.

[0052] 18. Item 18: The material of item 16 or 17, wherein said substituent is on a sp2 carbon atom.

[0053] 19. Item 19: The material of item 18, wherein the porphyrin-type macrocycle comprises an alkyl substituent on an sp2 carbon atom.

[0054] 20. Item 20: The material of item 19, wherein the porphyrin-type macrocycle comprises an alkyl substituent on an sp2 carbon atom selected from C1, C4, C8, and C10.

[0055] It is observed that the choice of the substituents on the carbon of the macrocycle influence the antiviral efficiency of the material.

[0056] 21. Item 21 : The material of any one of items 14 to 20, wherein the covalent link between the surface oxygen atoms O1and the porphyrin-type macrocycle comprises a CH2-CH2 moiety directly linked to the porphyrin-type macrocycle. Without wishing to be bound by theory, it is believed that flexibilizing the linker facilitates a high functionalization of the material.

[0057] As defined herein, the CH2-CH2 moiety is part of the photosensitizer P.

[0058] 22. Item 22: The material of any one of items 14 to 21, wherein the covalent link between the surface oxygen atoms O1and the porphyrin-type macrocycle is linked to a carbon atom of the porphyrin-type macrocycle selected from C1, C2, C4, C5, C7, C8, C10, and C11, preferably selected from C5and C7.

[0059] 23. Item 23: the material of any one items 14 to 22, wherein the porphyrin type macrocycle is a chlorin type macrocycle.

[0060] As defined herein a chlorin-type macrocycle is a molecule comprising moiety II wherein at least one of {C1, C2}, {C4, C5}, and {C7, C8} comprises two sp3 hybridized carbons, or the corresponding metalated moiety.

[0061] 24. Item 24: the material of item 23, wherein either {C1, C2} or {C7, C8} comprises two sp3 hybridized carbons.

[0062] 25. Item 25: the material of item 24, wherein the chlorin-type macrocycle comprises moiety III, or the corresponding metalated moiety, wherein any of C'-C12may bear at least one substituent and wherein C7and C8are sp3 hybridized.

[0063] 26. Item 26: the material of item 25, wherein the chlorin-type macrocycle comprises moiety IV, or the corresponding metalated moiety, wherein any of C'-C4, C7-C12, Caand Cbmay bear at least one substituent, wherein C7and C8are sp3 hybridized and wherein Caand Cbmay be sp2 or sp3 hybridized. 27. Item 27: the material of item 26, wherein the chlorin type macrocycle comprises moiety V, or the corresponding metalated moiety, wherein any of C'-C4, C7-C12, and Cbmay bear at least one substituent, wherein C7, C8and Cbare sp3 hybridized.

[0064] 28. Item 28: the material of item 27, wherein the chlorin type macrocycle is a pheophorbide derivative of formula VI or the corresponding metalated moiety, which is covalently linked to the surface oxygen atoms O1via R.

[0065] VI

[0066] 29. Item 29: the material of any one of items 1 to 28, wherein P is a ligand and is non metalated.

[0067] 30. Item 30: the material of any one of items 1 to 29, wherein the covalent link between O1and P comprises the following moiety: O’-Si-C

[0068] 31. Item 31: the material of item 30, wherein the covalent link between O1and P comprises the following moiety: CF-Si-CHz-CHz-CHz-N

[0069] 32. Item 32: the material of item 31, wherein P comprises a pheophorbide derivative of formula VI and wherein the covalent link between O1and P comprises the moiety: O’-Si-CHz-CHz-CHz-NH, wherein the nitrogen atom is directly linked to the carbon atom linked to R in formula VI.

[0070] Accordingly, it should be understood that in the material of item 24, R comprises the moiety Si-CHz-CHz-CHz-

[0071] NH. 33. Item 33: A composition comprising the material of any one of items 1 to 32 for a use in a therapeutic method of treatment of a disease caused by an enveloped virus.

[0072] The therapeutic method of treatment is not particularly limited. It may include ingesting the composition or injecting the composition in or applying the composition to a part of the body to be treated.

[0073] The composition may comprise the material in the form of nanoparticles dispersed in the composition.

[0074] The composition of the invention may consist of the material of the invention. It may also comprise a solvent, such as water, in which the material may be dispersed in the form of particles, e.g. nanoparticles. The composition of the invention may comprise pharmaceutically acceptable additives and excipients.

[0075] 34. Item 34: the composition of item 33, wherein the enveloped virus is a virus of the Coronaviridae family or a virus of the Flaviviridae family.

[0076] 35. Item 35: the composition of item 34, wherein the enveloped virus is a virus of the genera Alphacoronavirus, Betacoronavirus, or Hepacivirus.

[0077] 36. Item 36: the composition of item 35, wherein the enveloped virus is a virus selected from HCoV-229E, SARS-CoV-2, MERS-CoV and HCV.

[0078] 37. Item 37: the composition of any one of items 33 to 36, wherein the treatment includes injecting the composition in or applying the composition to a part of the body to be treated and exposing of the material to light.

[0079] Exposing the material to light may comprise delivering light from daylight or from a light source, such as a lamp or a laser, to the material by use of an optical fiber.

[0080] 38. Item 38: A coating composition comprising the material of any one of items 1 to 32.

[0081] The coating composition of the invention is not particularly limited. It may comprise a binder and / or a solvent, in addition to the material of the invention. The binder and the solvent and additives present in the composition are not particularly limited and can be chosen based on the type of coating desired and the surface to be coated.

[0082] 39. Item 39: A coated object wherein the coating comprises the material of any one of items 1 to 32.

[0083] Said object is not particularly limited. It could be a filter, for example a face mask or a filter for decontaminating air or water.

[0084] 40. Item 40: A method of preparation of the coated object of item 39 comprising the following steps: Providing a non-coated object, applying the coating composition of the invention on the non-coated object, obtaining the coating from the applied coated composition.

[0085] The non-coated object is not particularly limited, it may include fibers or fibrous materials such as yarn, woven materials, or non-woven materials. It may include also non-fibrous materials, such as a glass sheet or a plastic sheet.

[0086] The method of applying the coating composition is not particularly limited. It may include all coating techniques well known in the art. The method of obtaining the coating from the applied coating composition is not particularly limited. Depending on the amount and nature of binder and / or solvent present in the coating composition, it may comprise various curing and / or drying methods.

[0087] 41. Item 41: Non-therapeutical use of the material of any one of items 1 to 32, of the coating composition of item 38, or of the coated object of item 39, to deactivate an enveloped virus.

[0088] Such use can include applying light (e.g. daylight) to an object comprising the material of the invention to enhance the antiviral properties of the object and decontaminate the surface of the object. It may comprise the use of the material to decontaminate fluids or surfaces contaminated with enveloped viruses, or the treatment of plants infected with an enveloped virus.

[0089] 42. Item 42: the use of item 41, wherein the enveloped virus is a virus of the Coronaviridae family or a virus of the Flaviviridae family.

[0090] 43. Item 43: the use of item 42, wherein the enveloped virus is a virus of the genera Alphacoronavirus, Betacoronavirus, or Hepacivirus.

[0091] 44. Item 44: the use of item 43, wherein the enveloped virus is a virus selected from HCoV-229E, SARS- CoV-2, MERS-CoV and HCV.

[0092] 45. Item 45: the use of any one of items 41 to 44, wherein the deactivation comprises the use of P to produce a reactive oxygen species and the reaction of the reactive oxygen species with a lipid of the envelope of the enveloped virus.

[0093] 46. Item 46: the use of any one of items 41 to 45, wherein the use comprises contacting the enveloped virus and the material.

[0094] As defined herein, if the material is in contact with a fluid, in which the virus is suspended, it is considered to be in contact with the virus.

[0095] 47. Item 47: the use of any one of items 41 to 46, wherein the use comprises exposing the material to light when the material is in contact with the enveloped virus.

[0096] Exposing the material to light may comprise delivering light from a light source, such as a laser, to the material by use of an optical fiber.

[0097] In some embodiments of the non-therapeutical use of the material, exposing the material to light consist of an exposure of the material to daylight. It should be understood that exposure of the material to daylight comprises exposure to daylight through a transparent or translucent medium.

[0098] It has been observed that, with the material of the invention it is not necessary to use a light source other than daylight to obtain an antiviral effect.

[0099] 48. Item 48 : A process for the preparation of the material of any one of items 1 to 32, comprising the following steps: a. Providing the inorganic oxide or hydroxide comprising surface oxygen atoms O'; b. Providing the photosensitizer P; c. Covalently linking P to O1to obtain the material. 49. Item 49: the process of item 48, wherein step c. comprises: i. reacting O1with a molecule bearing a function F able to react with P to obtain a functionalized inorganic oxide or hydroxide, wherein O1is linked to F via one or more covalent bonds, and ii. reacting P with F to form a covalent bond between P and F to obtain the material.

[0100] F designates a functional group and any protonated or deprotonated form of said functional group.

[0101] 50. Item 50: the process of item 49, wherein the molecule bearing a function F is a silicon alkoxide comprising the moiety: AlkO-Si-X-F, X being an alkylene radical, and wherein the product of the reaction of O1with the molecule bearing a function F comprises the moiety: O’-Si-X-F.

[0102] For example, the reaction for obtaining O’-Si-X-F from AlkO-Si-X-F and the inorganic oxide or hydroxide may consist of heating the molecule bearing a function F and the inorganic oxide or hydroxide in an organic solvent.

[0103] 51. Item 51: the process of item 49 or 50, wherein F is an amine function and P bears a carboxyl group and wherein the reaction between P and F comprises the formation of an amide by condensation of the amine function and the carboxyl group.

[0104] For example, a coupling reaction using the coupling agent l-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDO) may be used for the formation of the amide. Preferably, hydroxybenzotriazole (HOBt) or N- hydroxysuccinimide (NHS) are also used in said coupling reaction.

[0105] 52. Item 52: the process of item 51, wherein the molecule bearing the amine function is a (3- aminoalkyl)trialkoxysilane, for example (3-aminopropyl)triethoxysilane.

[0106] 53. Item 53: the process of any one of items 48 to 52, wherein step a. comprises: iii. Providing a precursor of the inorganic oxide or hydroxide, and iv. Obtaining the inorganic oxide or hydroxide from the precursor by condensation of the precursor in a sol-gel process.

[0107] 54. Item 54: the process of item 53, wherein the sol-gel process is acid catalyzed.

[0108] 55. Item 55: the process of item 53 or 54, wherein the condensation is performed in the presence of a templating material, which is removed after completion of the condensation to obtain a porous inorganic oxide.

[0109] 56. Item 56: the process of item 55, wherein the templating material is a poloxamer.

[0110] 57. Item 57: the process of any one of items 53 to 56, wherein the sol-gel process further comprises the calcination of the product of the condensation of the precursor to obtain the inorganic oxide.

[0111] 58. Item 58: the process of any one of items 53 to 57, wherein the precursor is an alkoxide of the element E.

[0112] 59. Item 59: the process of item 58, wherein the precursor comprises a tetraalkoxysilane, preferably the precursor is tetraethoxysilane.

[0113] Examples

[0114] Materials Zirconium(IV) chloride (ZrC14, >99.5%), benzoic acid (99%), 1,4-benzenedicarboxylic acid (H2BDC, 98%), 2- amino-4-benzenedicarboxylic acid (H2N-BDC, 99%) and anhydrous N-N-dimethylformamide (DMF) were purchased from Alfa Aesar. Hydrochloric acid 37% aluminum(III) chloride hexahydrate (A1C13,6 H2O, >99.0%) and formic acid were bought from Fisher scientific. Acetone and dichloromethane were bought from VWR. N-N- dimethylformamide (DMF) was bought from Carlo Erba. Methanol was bought from Brabant and anhydrous methanol was prepared by putting 50 g of molecular sieves 4 A in 250 mL of methanol. Titanium tetraisopropoxide 97% (TTIP), tetraethyl orthosilicate (TEOS), (3-aminopropyl)triethoxysilane (APTES), Propionic acid (>99.5%), l-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride (EDCI’HCl) and 1 -hydroxybenzotriazole hydrate (HOBt) were purchased from Sigma-Aldrich. 5,10,15,20-(tetra-4-carboxyphenyl)porphyrin (TCPP, >98%) was purchased from PorphyChem. Pheophorbide A (PhA) was bought from Cayman Chemicals. Dulbecco’s phosphate buffered saline (DPBS or PBS) IX, Dulbecco’s Modified Eagle Medium (DMEM) IX, Trypsin-EDTA 0.05X were bought from Gibco. Fetal Bovine Serum (FBS) was bought from Eurobio. Neutral Red (NR) was bought from Clin-tech. Renilla Lysis buffer and the Luciferase reagent were bought from Promega.

[0115] Huh7, VeroE6 and Vero81 (ATCC number CCL-81) were grown in DMEM with 10% FBS in an incubator at 37 °C with 5% CO2. Vero81 cells were subcloned to obtain a better overall infection rate. HCoV-229E strain VR-740 (ATCC) and recombinant HCoV-229E-Luc were used. SARS-CoV-2 (isolate SARS-CoV- 2 / human / FRA / Lille_Vero-81-TMPRSS2 / 2020; NCBI MW575140) was propagated on Vero81 TMPRSS2 cells.

[0116] Synthesis of Al-TCPP, PCN-222, PCN-223, PCN-224, SBA-15, SBA-15-NH2, UiO-66(Zr), UiO-66(Zr)-NH2, MIL-125(Ti), MIL-125(Ti)-NH2

[0117] Al-TCPP, PCN-222, PCN-223, and PCN-224 are MOFs comprising TCPP (a heterocyclic macrocyclic photosensitizer) as organic building block.

[0118] Al-TCPP (comparative) was synthesized by a solvothermal method. 150 mg of A1C13*6H2O along with 200 mg of TCPP were added in a 100 mL Teflon lined autoclave that contained 25 mL of deionized water and heated at 180 °C for 16 h. After being cooled down to room temperature, the suspension was centrifuged at 4000 rpm for 15 min and the crystals were washed two times with 50 mL of DMF (2x50 mL) and two times with 50 mL of acetone (2x50 mL). Lastly, the resulting product was put in a vacuum oven at 100 °C overnight to evaporate any remaining solvent.

[0119] PCN-222 (comparative) was prepared by dissolving 42 mg of ZrC14 and 60 mg of TCPP in an Erlenmeyer flask that contained 60 mL of DMF and 16.92 mL of formic acid. Afterwards, the solution was transferred to a 125 mL Teflon lined autoclave and heated at 120 °C for 16 h.

[0120] PCN-223 (comparative) was synthesized by adding 38 mg of ZrC14 and 50 mg of TCPP in an Erlenmeyer flask and subsequently 50 mL of DMF and 10 mL of propionic acid were put in the flask. After dissolving the reagents, the solution was transferred to a 100 mL Teflon lined autoclave and heated at 130 °C for 16 h.

[0121] PCN-224 (comparative) was produced by dissolving 90 mg of ZrC14, 30 mg of TCPP and 1.2 g of benzoic acid in 6 mL of DMF in a 25 mL Teflon lined autoclave, and then heating the solution at 120 °C for 24 h.

[0122] The three TCPP MOFs were centrifuged, after being cooled down, at 4000 rpm for 15 min. The products were obtained and washed three times with 20 mL of DMF (3x20 mL) and three times with 20 mL of acetone (3x20 mL). Then, the products were soaked in 20 mL of acetone during 24 h as a final step of washing and finally they were dried in a vacuum oven at 100 °C overnight.

[0123] SBA-15 (comparative) was prepared as follows. Firstly, 7.5 L of 1.6 M HC1 were put in a 10 L glass reactor, along with 200 g of Pluronic P123 and stirred for 24 h at 40 °C at 160 rpm to completely dissolve the copolymer. The second day, 480 mL of TEOS were added with the use of a Hei-flow precision pump regulated at 50 rpm, in order to allow the slow addition of the silica precursor inside the solution. The solution was stirred for another 24 h at the same temperature and speed. Afterwards, the solution is heated at 100 °C during 48 h under stirring to allow the reaction to take place. The fifth day, the solid that is formed is filtered by Buchner and it is further washed with distilled water. Then, the product is dried overnight at 70 °C and lastly calcined at 500 °C for 6 h, with a step of 1 °C / min so as to remove the copolymer and form the mesopores of the silica. After calcination, SBA-15 was stored in a desiccator with 0% relative humidity.

[0124] SBA-15-NH2 (comparative) was synthesized as follows. 0.5 g of calcined SBA-15 were dried in an oven at least for 3 h at 100 °C before the synthesis. Then, the dried SBA-15 was placed in a round spherical flask, along with a small magnetic stirring rod, 40 mL of toluene and 10 mL of APTES. The flask was put in an oil bath on top of a magnetic stirrer and the reaction was done under reflux and under stirring (500 rpm) during 24 h. Once the reflux was done, the product was collected by filtration and the functionalized SBA-15 was washed with toluene (3x10 mL) and finally dried at 60 °C overnight. SBA-15-NH2 was also stored in a desiccator, to avoid the hydrolysis of the amino group.

[0125] UiO-66(Zr) and UiO-66(Zr)-NH2 are MOF having the same structure, the only difference is that in UiO-66(Zr) the organic linker is BDC and in UiO-66(Zr)-NH2 the organic linker is HjN-BDC.

[0126] UiO-66(Zr) (comparative) As described in Plastiras, O.-E. et al., Cytotoxicity and effectiveness of archetypal Metal-Organic Frameworks (HKUST-1, UiO-66, MIL-53, MIL-125) against coronaviruses (HCoV-229E and SARS-CoV-2). Microporous Mesoporous Mater. 2023. DOI: 10.1016 / j.micromeso.2023.112975.

[0127] UiO-66(Zr)-NH2 (comparative) was prepared by a solvothermal method. 4 g of ZrC14 and 6 g of BDC-NH2 were put in a 1 L glass reactor, which contained 650 mL of DMF and 32 mL of formic acid. The mixture is allowed to be stirred for 30 minutes so as to completely dissolve the reactants and then the reactor was transferred to an oven at 120 °C for 24 h. Then, the reactor was left to cool down and the solid product was separated by centrifugation at 4000 rpm for 15 min. The product was collected, washed three times with DMF (3x80 mL) and three one-day cycles with methanol (3x80 mL per day). Lastly, the product was dried in a vacuum oven at 100 °C overnight.

[0128] MIL-125(Ti) and MIL-125(Ti)-NH2 are MOF having the same structure, the only difference is that in MIL-125(Ti) the organic linker is BDC and in MIL-125(Ti)-NH2 the organic linker is HjN-BDC.

[0129] MIL-125(Ti) (comparative) As described in Plastiras, O.-E. et al., Cytotoxicity and effectiveness of archetypal Metal-Organic Frameworks (HKUST-1, UiO-66, MIL-53, MIL-125) against coronaviruses (HCoV-229E and SARS-CoV-2). Microporous Mesoporous Mater. 2023. DOI: 10.1016 / j.micromeso.2023.112975.

[0130] MIL-125(Ti)-NH2 (comparative) was synthesized inside a glove box, by firstly drying a 125 mL Teflon line autoclave in an oven at 100 °C overnight. The following day, the autoclave was inserted into the glove box and 28.4 mL of anhydrous DMF with 2.64 g of BDC-NH2 are put in the autoclave and stirred until dissolution. Subsequently, 3.16 mL of anhydrous methanol and 1.75 mL of TTIP are put dropwise onto the solution and stirred for 10 min. 25.24 mL of anhydrous methanol are added and the autoclave is heated at 150 °C for 16 h. The solid product was, once more, separated by centrifugation and it was washed thoroughly by immersing it in 30 mL of DMF and performing three one-day cycles (3x30 DMF per day) and then immersing it in 30 mL of methanol again for three one-day cycles (3x30 MeOH per day). Finally, the yellow MOF is dried in a vacuum oven at 120 °C overnight.

[0131] Functionalization of SBA-15-NH2, UiO-66-NH2, MIL-125(Ti)-NH2 with PhA

[0132] SBA-15-NH-PhA (inventive) was synthesized by covalently binding Pheophorbide A onto the silica via a coupling between the carboxyl function of the porphyrin and the amino group of the mesoporous silica, resulting in an amide. 0.25 g of SBA-15-NH2 were added in a Schlenk flask, along with 25 mg of PhA, 15 mg of EDCI’HCI that serves as the coupling agent and 7.5 mg of HOBt as a catalyst. 25 mL of dry CH2C12 and a magnetic stirring rod were added to the flask and the suspension was left to react during 24 h at room temperature under stirring (700 rpm) and under inert atmosphere. Next, the suspension was filtered and washed two times with 10 mL of CH2C12 (2x10 mL) to remove any unreacted reagents. Then, the green product was dried in an oven at 50°C overnight, stored in a vial and kept in the dark.

[0133] UiO-66(Zr)-NH2 and MIL-125(Ti)-NH2 were functionalized with Pheophorbide A using the same procedure as for the SBA-15-NH2. Briefly, 26 mg of UiO-66(Zr)-NH2 or 25 mg of MIL-125(Ti)-NH2 were put in a Schlenk flask that contained 10 mg of PhA, 6 mg of EDCI’HCI and 3 mg of HOBt in 10 mL of dry CH2C12. The reaction was left to take place for 24 h at room temperature and then the two suspensions were centrifuged, and the two functionalized MOFs were washed two times with 20 mL of dry CH2C12. Lastly, the greenish UiO-66(Zr)-NH- PhA (comparative) and the yellow-green MIL-125(Ti)-NH-PhA (comparative) were dried in an oven at 50 °C overnight, stored in a vial and kept in the dark.

[0134] Functionalization of SBA-15-NH2 with Chlorin e6 (Ce6), Tetrakis(4-carboxyphenyl)porphyrin (TCPP) and Protoporphyrin IX (PPIX)

[0135] SBA-15-NH-Ce6, SBA-15-NH-TCPP and SBA-15-NH-PPIX were synthesized with the same method as the one used with Pheophorbide, by covalently binding the photosensitizers onto the silica via a coupling between the carboxyl function of the chlorin or porphyrin and the amino group of the mesoporous silica, resulting in an amide. 0.1 g of SBA-15-NH2 were added in a Schlenk flask, along with 10 mg of Ce6, TCPP or PPIX, 6 mg of EDCI’HCI that activates the carboxylic group and 4 mg of HOBt as a coupling agent. 10 mL of dry CH2Q2 and a magnetic stirring rod were added to the flask and the suspension was left to react during 24 h at room temperature under stirring (700 rpm) and under inert atmosphere (N2). Next, the suspension was filtered and washed three times with 10 mL of CH2Q2 (2x10 mL) to remove any unreacted reagents. Then, the green (SBA-15-NH-Ce6) or red (SBA- 15-NH-TCPP and SBA-15-NH-PPIX) resulting product was dried in an oven at 50 °C overnight, stored in a vial and kept in the dark, by wrapping it in aluminum foil.

[0136] TCPP has the following structure:

[0137]

[0138] Ce6 has the following structure:

[0139] PPIX has the following structure:

[0140] TGA Analysis

[0141] Thermogravimetric analyses (TGA) were performed on SBA-15-NH-PhA, SBA-15-NH-Ce6, SBA-15-NH-TCPP and SBA-15-NH-PPIX with a Mettler Toledo TGA / DSC 3+ thermal analyzer in the range of 100-1000 °C under an air flow of 80 mL / min, with a heating rate of 5 °C / min. A mass loading percentage in the macrocycles for each functionalized silica was calculated from the observed weight losses using the following formula: Where Aw is the weight loss at 200 °C or 250 °C minus the one at 600 °C, f is for functionalized and nf stands for non-functionalized with a macrocycle. It was found that SBA-15-NH-PhA was functionalized with 6.85% wt. of PhA, SBA-15-NH-Ce6 was functionalized with 8.33 wt.% of Ce6, SBA-15-NH-TCPP was functionalized with 7.21 wt.% of TCPP, and SBA-15-NH-PPIX was functionalized with 10.67 wt.% of PPIX. Thus SBA-15-NH-TCPP contained 10 mmol TCPP per 100 g of material, SBA-15-NH-PhA contained 12 mmol PhA per 100 g of material, SBA-15-NH-Ce6 contained 15 mmol Ce6 per 100 g of material, and SBA-15-NH-PPIX contained 21 mmol PPIX per 100 g of material.

[0142] Cell culture and cytotoxicity assay

[0143] The materials were evaluated for their cytotoxicity with the Neutral Red method (NR) against Huh7 TMPRSS2 and Vero81.6 cells. The preparation of the different concentrations of the materials, the protocol of the assay and the cell culture were done as described in Plastiras, O.-E. et al., Cytotoxicity and effectiveness of archetypal Metal- Organic Frameworks (HKUST-1, UiO-66, MIL-53, MIL-125) against coronaviruses (HCoV-229E and SARS- CoV-2). Microporous Mesoporous Mater. 2023. DOI: 10.1016 / j.micromeso.2023.112975.

[0144] Infection tests with HCoV-229E, SARS-CoV-2, MERS-CoV and HCV

[0145] The assessment of the antiviral activity of the materials against HCV was done as described in de Fourchambault, E. M.; et al. Hepatitis C virus alters the morphology and function of peroxisomes. Frontiers in Microbiology 2023, 14. DOI: 10.3389 / fmicb.2023.1254728.

[0146] The assessment of the antiviral activity of the materials against HCoV-229E and SARS-CoV-2 was done as described in Plastiras, O.-E. et al., Cytotoxicity and effectiveness of archetypal Metal-Organic Frameworks (HKUST-1, UiO-66, MIL-53, MIL-125) against coronaviruses (HCoV-229E and SARS-CoV-2). Microporous Mesoporous Mater. 2023. DOI: 10.1016 / j.micromeso.2023.112975.

[0147] For HCoV-229E in vitro antiviral activity of the non-functionalized and functionalized porous materials was assessed against HCoV-229E-Luc, a luciferase recombinant virus of HCoV-229E, in Huh7 TMPRSS2 cells.

[0148] For MERS-CoV, a similar procedure as SARS-CoV-2 was followed as described below.

[0149] The day before the infection test with MERS-CoV, a 24-well plate was prepared with 125,000 cells / well of Huh7- AP and incubated at 37 °C, 5% CO2 for 24 h. The day of the infection, the tubes containing 0.75 mg of the material to be tested are prepared, by adding 1.5 mL of DMEM + 10% of FBS, containing the virus at a multiplicity of infection (MOI) of 0.1, thus achieving a concentration of 0.5 mg / mL of the material. The tube containing the material and the virus is incubated at room temperature under light irradiation for 15 min, and then it is centrifuged at 5000 rpm for 5 min. 0.5 mL of the supernatant was added to 2 of the wells of the 24-well plate, while 2 wells were infected with 0.5 mL of the virus (negative control) and another 2 wells with 0.5 mL of complete medium containing 5 LI of remdesivir (positive control).

[0150] Later, the 24-well plate was incubated at 37 °C, 5% CO2 for 1 h, and the medium of the wells was refreshed, by removing the old and adding the new one at the same volume. Afterwards, the 24-well plate was further incubated at 37 °C, 5% CO2 for 16 h and the supernatants were collected to proceed with the TCID50 method. For the immunofluorescence (IF) method, the microscopy glass slides were fixed two times with PFA4% for 30 min each, by changing the plate between each fixation.

[0151] For the first method (TCID50 method), 96-well plates having 10,000 cells / well of Huh7-AP cells were prepared and incubated at 37 °C, 5% CO2 for 24 h. Next, the supernatants collected after the infection were diluted from 10-1 to 10-6 and added to the 96-well plates. The titer was measured at three different days (day 3, 5 and 7) to check the effect of cytopathy and the TCID50 was calculated by the formula of Spearman and Karber.

[0152] For the second method (IF method), the glass slides were washed three times with PBS IX and submerged in PBS IX + 0.4% Triton for 3 min. Next, another three washes with PBS IX took place and the slides were blocked by using 5% of goat serum for 30 min. The slides were incubated with 0.3-0.5 mL of primary antibodies (anti-double strand and anti-S protein of MERS). The slides were washed three times with PBS IX and then they were incubated with 0.3-0.5 mL of secondary antibodies (A488-GAM, A594-GAR and DAPI). After 3 washes of PBS IX, the slides were mounted on slats with 7 LIL of Mowiol®. 10 images were taken with the 10X lens of Evos M5000 (Thermo Fischer Scientific) and analyzed by the software Fiji.

[0153] The antiviral activity of the materials was tested at different time points and both under dark, by wrapping the tubes with aluminum foil, and under visible light irradiation, by the exposure of the BSL’s lamp (fluorescent tube, 36 W, 3350 lumens, white light).

[0154] Results

[0155] Table 1 summarizes the results of a first batch of experiments. Unless otherwise indicated, in table 1, the antiviral activities correspond to the activities measured for 15 minutes of exposure of the virus to 1 mg / mL of the material.

[0156] Cytotoxicity

[0157] The materials were studied for their cytotoxicity with the Neutral Red assay against Huh7 TMPRSS2 and Vero81.6 cells. The cytotoxicity indicated in table 1 is at the highest safest concentration (1 mg / mL for all materials except for MIL-125(Ti) and Al-TCPP for Huh7 TMPRSS2 cells where the concentration was 0.1 mg / mL and 0.5 mg / mL respectively). None of the materials tested showed cytotoxicity at these concentrations.

[0158] HCoV-229E

[0159] The non-functionalized materials, SBA-15, SBA-15-NH2, UiO-66(Zr), UiO-66(Zr)-NH2, MIL-125(Ti), MIL- 125 (Ti)-NH2, PCN-222, PCN-223, and PCN-224 and Al-TCPP were studied at 60 min of contact with the virus. In the case of MIL-125(Ti) and Al-TCPP, the concentration of the material was reduced to 0.1 mg / mL and 0.5 mg / mL respectively due to its higher cytotoxicity.

[0160] SARS-CoV-2 UiO-66(Zr), MIL-125(Ti), MIL-125(Ti)-NH2, Al-TCPP, PCN-222, SBA-15-NH-PhA, UiO-66(Zr)-NH-PhA and MIL-125(Ti)-NH-PhA were tested against SARS-CoV-2 both under light irradiation and in the dark. For UiO- 66(Zr), MIL-125(Ti) and MIL- 125(Ti)-NH2the concentration of the material was 5 mg / mL.

[0161] MERS-CoV and HCV SBA-15-NH-PhA, Al-TCPP and UiO-66(Zr)-NH-PhA were further studied for their antiviral activity against MERS-CoV and HCV, inactivating them under 15 min of light exposure.

[0162]

[0163] Table 1. Table 2 summarizes the results of a second batch of experiments concerning SBA-15-NH-Ce6, SBA-15-NH-TCPP and SBA-15-NH-PPIX. The antiviral activities correspond to the activities measured with exposure to light for 10 minutes of exposure of the virus to 1 mg / mL of the material.

[0164] SBA-15-NH-Ce6, SBA-15-NH-TCPP and SBA-15-NH-PPIX showed no cytotoxicity versus Huh7-TMPRSS2 cells up to the highest concentration tested (1 mg / mL).

[0165] Table 2.

Claims

CLAIMS1. Material comprising an outside surface and an inorganic oxide or hydroxide of an element E, said outside surface comprising a photosensitizer P, and said inorganic oxide or hydroxide comprising surface oxygen atoms O1covalently linked to said photosensitizer.

2. The material of claim 1, wherein the inorganic oxide or hydroxide is obtained by a sol-gel process comprising the condensation of a precursor of the inorganic oxide or hydroxide.

3. The material of claim 2, wherein the sol-gel process comprises the calcination of the product of the condensation.

4. The material of claim 2 or 3, wherein the sol-gel process is acid catalyzed.

5. The material of any one of claims 1 to 4, wherein the photosensitizer P is a porphyrin-type macrocycle comprising moiety II or the corresponding metalated moiety:IIWherein C3, C6, C9, and C12are directly linked to an atom selected from H and a sp3 carbon atom.

6. The material of claim 5, wherein the covalent link between the surface oxygen atoms O1and the porphyrin-type macrocycle comprises a CH2-CH2 moiety directly linked to the porphyrin-type macrocycle.

7. A composition comprising the material of any one of claims 1 to 6 for a use in a therapeutic method of treatment of a disease caused by an enveloped virus.

8. A coating composition comprising the material of any one of claims 1 to 6.

9. A coated object wherein the coating comprises the material of any one of claims 1 to 6.

10. A method of preparation of the coated object of claim 9 comprising the following steps: o providing a non-coated object, o applying the coating composition of the invention on the non-coated object, o obtaining the coating from the applied coated composition.

11. Non-therapeutical use of the material of any one of claims 1 to 6, of the coating composition of claim 8, or of the coated object of claim 9, to deactivate an enveloped virus.

12. A process for the preparation of the material of any one of claims 1 to 6, comprising the following steps: a. Providing the inorganic oxide or hydroxide comprising surface oxygen atoms O'; b. Providing the photosensitizer P; c. Covalently linking P to O1to obtain the material.

13. The process of claim 12, wherein step a. comprises:iii. Providing a precursor of the inorganic oxide or hydroxide, and iv. Obtaining the inorganic oxide or hydroxide from the precursor by condensation of the precursor in a sol-gel process.

14. The process of claim 13, wherein the sol-gel process is acid catalyzed.

15. The process of claim 13 or 14, wherein the sol-gel process further comprises the calcination of the product of the condensation of the precursor to obtain the inorganic oxide.

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