A FATTY ACID BASED COMPOSITION FOR TREATMENT AND/OR PREVENTION OF ENVELOPED-VIRUS RELATED INFECTIONS

Abstract

A fatty acid based composition for treatment and/or prevention of enveloped-virus related infections by administering fatty acid based compositions. The present invention further provides a free fatty acid based sprayable composition to prevent and/or treatment of SARS-Virus related infections. The fatty acids used in the present invention are Linoleic acid (LA), α-linolenic acid (ALA) and gamma-linolenic acid (GLA). In a preferred embodiment, we propose that topical administration of naturally occurring fatty acids in an optimal formulation may be beneficial in the prophylaxis of respiratory enveloped viruses and in particular the lipid-rich SARS-CoV-2 virus. This could be administered via nasal and/or oral route. The composition comprises at least one fatty acid, a suitable carrier, preservative, cosolvents and surfactants and/or cyclodextrin used as an excipient that may also enhance anti-viral properties.

Claims

1. An antiviral composition comprising an effective amount of one or more fatty acid a suitable carrier, a preservative, one of more cosolvents, and one or more surfactants; wherein, the composition comprises between 0.01% and 10% fatty acid; the composition is a disinfectant composition comprising a pH in the range of 5-7.5; and the antiviral activity is for enveloped virus including SARS-CoV-2.

2. The composition as claimed in claim 1, wherein the at least one fatty acid is selected from the group of LA, GLA, DGLA, Oleic acid, ALA, EPA, ARA, DHA, caprylic acid, lauric acid or any other Poly-Unsaturated fatty acid and amphipathic fatty acids.

3. The composition as claimed in claim 1, wherein the composition is in the form of a spray, emulsion, suspension, solution, foam, ointment, liquid soap, cream, mouthwash, jelly, lozenge or pastille.

4. The composition as claimed in claim 1, wherein the composition is in the form of nasal spray, mouth spray and topical ointment.

5. The composition as claimed in claim 1, wherein the composition is mucoadhesive.

6. A method of preventing and/or treating microbial infection including those caused by enveloped viruses by administering a fatty acid based composition.

7. The method as claimed in claim 6, wherein the method inhibits the growth of enveloped viruses by rupturing lipid-rich protein outer envelope coating of the virus.

8. The method as claimed in claim 7, wherein the method inhibits the growth of SARS-CoV-2 virus.

9. The method as claimed in claim 7, wherein the method is used to disinfect the surfaces and other fomites.

10. The method as claimed in claim 7, wherein the composition is administered topically.

11. An antiviral composition comprising an effective amount of one or more fatty acid, a suitable carrier, a preservative, one of more cosolvents, and one or more surfactants; wherein, one or more free fatty acids is selected in the ratio of about 5:1 to about 1:5; the composition comprises between 0.01% and 10% free fatty acid; the composition is a disinfectant composition comprising a pH in the range of 5-7.5; and the antiviral activity is for enveloped virus including SARS-CoV-2.

12. The composition as claimed in claim 11, wherein the at least one fatty acid is selected from the group of LA, GLA, DGLA, Oleic acid, ALA, EPA, ARA, DHA, caprylic acid, lauric acid or any other fatty acid; and/or their monoglycerides.

13. The composition as claimed in claim 11, wherein the composition is in the form of a spray, emulsion, suspension, solution, foam, ointment, liquid soap, cream, mouthwash, jelly, lozenge or pastille.

14. The composition as claimed in claim 11, wherein the composition is in the form of nasal spray, mouth spray and topical ointment.

15. The composition as claimed in claim 11, wherein the composition is mucoadhesive.

16. The composition as claimed in claim 11, wherein the fatty acids mixed with cyclodextrin as a preferred surfactant.

17. Use of the composition as claimed in claim 11, in food processing industry including but not limited to sanitizing all kinds of surfaces, packaging, table wares, eatables etc., especially where soaps and bleach based disinfectants are not recommended.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0042] An understanding of the antiviral composition and method of preventing and/or treating microbial infection of the present invention may be obtained by reference to the following drawings:

[0043] FIG. 1 graphically represents percentage reduction in viral load by using the composition containing LA.

[0044] FIG. 2 graphically represents percentage reduction in viral load by using the composition containing GLA.

[0045] FIG. 3 shows the particle size of ALA using Malvern zetasizer. The mean particle size obtained using the present composition was 170.4 nm when the sample was diluted 50 times.

[0046] FIG. 4 shows the particle size of GLA using Malvern zetasizer. The mean particle size obtained using the present composition was 283.2 nm when the sample was diluted 50 times.

[0047] FIG. 5 shows the particle size of LA using Malvern zetasizer. The mean particle size obtained using the present composition was 183 nm when a sample was diluted 50 times.

DETAILED DESCRIPTION OF THE INVENTION

[0048] The details of one or more embodiments of the invention are set forth in the accompanying description below including specific details of the best mode contemplated by the inventors for carrying out the invention. The embodiments of the invention which are apparent to one skilled in the art after reading the present disclosure and on applying the common general knowledge of the technical field are within the scope of this invention.

[0049] The present invention is based on the concept that enveloped viruses are protected by a fatty coating which is disrupted by detergents (usually on contact). This leads to destruction of the virus. Fatty acids are part of every cell and in food, have detergent actions. Fatty acid can inactivate/destroy viruses via their detergent actions. SARS-CoV-2 (Covid-19) is an enveloped virus and thus there is anticipated high probability that fatty acid can also inactivate SARS-CoV-2 (Covid-19) on contact.

[0050] The present invention uses fatty acids which are long chain carboxylic acid with long aliphatic chain. No double bond in the aliphatic chain forms saturated fatty acids. If one double bond is present in the aliphatic chain, it forms monounsaturated fatty acids. Two or more double bonds in the aliphatic chain forms polyunsaturated fatty acids (PUFA).

[0051] Fatty acids, primarily Linoleic acid (LA), α-linolenic acid (ALA) and gamma-linolenic acid (GLA) are taken for the study. It was seen that each of these fatty acids showed antiviral activity when incubated with enveloped viral cells, specifically SARS-CoV-2 cells. The fatty acid used in the present composition are used in purified or extracted form. Preferably, the composition used according to the present application comprises of a single fatty acid at a level that is of other fatty acids. Specifically, it implies that the other fatty acids are not present in a single composition.

[0052] Compositions comprising fatty acids have been found by the inventors to have broad antiviral activity. This activity provides wide application for the composition. It has also been found that composition can be used in different forms which further enhances the application of the composition. Since, the SARS-Co-2 virus enters the host via the nasal epithelium, therefore it is reasonable to expect that the treatment of such mucosal surfaces with fatty acids, such as linolenic acid etc. would result in the inactivation of SARS-CoV-2, thereby retarding or inhibiting the development of COVID-19.

[0053] If infection of the nasal passages may occur or has occurred or is desired to be prevented, administration of the composition to the nasal passages may be appropriate, and the composition may be in the form of a nasal ointment or nasal spray. If infection of the throat has occurred or is desired to be prevented, the composition may further be in the form of a throat gargle, lozenge, or spray and other forms of oral and nasal delivery.

[0054] In a preferred embodiment, the present invention provides an antiviral composition comprising an effective amount of one or more free fatty acids or free fatty acids (FA or FFA), a suitable carrier, a preservative, one of more cosolvents, and one or more surfactants;

wherein,
the composition comprises between 0.01% and 10% of fatty acid;
the composition is anti-microbial preparation with a pH in the range of 5-7.5; and
the antiviral activity is for enveloped virus including SARS-CoV-2.

[0055] In another preferred embodiment, the present invention provides a composition wherein at least one fatty acid is selected from but not limited to the group of LA, GLA, DGLA, Oleic acid, ALA, EPA, ARA, DHA, caprylic acid, lauric acid or any other fatty acids.

[0056] In yet another preferred embodiment, the present invention provides a composition that is in the form of a spray, emulsion, suspension, solution, foam, ointment, liquid soap, cream, mouthwash, jelly, lozenge or pastille.

[0057] In yet another preferred embodiment, the present invention provides a composition in the form of nasal spray, mouth spray and topical ointment.

[0058] In yet another preferred embodiment, the present invention discloses that a composition is mucoadhesive.

[0059] In yet another preferred embodiment of the present invention provides a method of preventing and/or treating microbial infection including those caused by enveloped viruses by administering a fatty acid based composition.

[0060] In yet another preferred embodiment, the present invention provides a method that inhibits the growth of enveloped viruses by rupturing lipid-rich protein outer envelope coating of the virus.

[0061] In yet another preferred embodiment, the present invention provides a method that inhibits the growth of enveloped viruses.

[0062] In yet another preferred embodiment, the present invention provides a method wherein the method inhibits the growth of SARS-CoV-2 virus.

[0063] In yet another preferred embodiment of the present invention provides a method used to disinfect the surfaces and other fomites.

[0064] In yet another preferred embodiment, the present invention provides a method wherein the composition is administered topically.

[0065] In yet another preferred embodiment, the present invention provides use of the composition in food processing industry including but not limited to sanitizing all kinds of surfaces, packaging, table wares, eatables etc., especially where soaps and bleach based disinfectants are not recommended.

[0066] In yet another preferred embodiment, the present invention provides a method of treating a disease or condition caused by SARS-CoV-2, the method comprising the step of administering to a subject in need thereof a composition disclosed in above embodiments.

[0067] In yet another preferred embodiment, the present invention provides use of the composition as disclosed in above embodiments, in food processing industry including but not limited to sanitizing all kinds of surfaces, packaging, table wares, eatables etc., especially where soaps and bleach based disinfectants are not recommended.

[0068] In still another preferred embodiment, the present invention provides an antiviral composition comprising an effective amount of one or more fatty acid, a suitable carrier, a preservative, one of more cosolvents, and one or more surfactants;

[0069] Wherein,

one or more free fatty acids is selected in the ratio of about 5:1 to about 1:5;
the composition comprises between 0.01% and 10% free fatty acid;
the composition is a disinfectant composition comprising a pH in the range of 5-7.5; and
the antiviral activity is for enveloped virus including SARS-CoV-2.

[0070] In another preferred embodiment, the present invention provides a composition that comprises at least one fatty acid, a suitable carrier, preservative, cosolvents, and surfactants. The proposed composition is in the form of spray. The surfactants in present fatty acid composition may be selected from polysorbate 20 (TWEEN 20), poly(ethylene glycol) 200 (PEG 200), Cyclodextrin, Triton and other suitable ether and anionic surfactants.

[0071] In yet another preferred embodiment, the present invention provides a composition wherein the at least one fatty acid is selected from the group of LA, GLA, DGLA, Oleic acid, ALA, EPA, ARA, DHA, caprylic acid, lauric acid or any other fatty acid; and/or their monoglycerides.

[0072] In yet another preferred embodiment, the present invention provides a composition wherein the composition is in the form of a spray, emulsion, suspension, solution, foam, ointment, liquid soap, cream, mouthwash, jelly, lozenge or pastille.

[0073] In yet another preferred embodiment, the present invention provides a composition wherein the composition is in the form of nasal spray, mouth spray and topical ointment.

[0074] In yet another preferred embodiment, the present invention provides a composition wherein the composition is mucoadhesive.

[0075] In yet another preferred embodiment, the present invention provides a composition which is one or more of the fatty acids mixed with cyclodextrin.

[0076] As shown in FIG. 1, graphically represents percentage reduction in viral load by using the composition containing LA.

[0077] As shown in FIG. 2, graphically represents percentage reduction in viral load by using the composition containing GLA.

[0078] As shown in FIG. 3, the particle size of ALA using Malvern zetasizer. The mean particle size obtained using the present composition was 170.4 nm when the sample was diluted 50 times.

[0079] As shown in FIG. 4, the particle size of GLA using Malvern zetasizer. The mean particle size obtained using the present composition was 283.2 nm when the sample was diluted 50 times.

[0080] As shown in FIG. 5, the particle size of LA using Malvern zetasizer. The mean particle size obtained using the present composition was 183 nm when a sample was diluted 50 times.

EXAMPLES

[0081] The following examples and advantages of the present invention are provided for the purpose of illustration only and are not intended to limit the scope of the present invention.

Example 1

Minimum Inhibitory Concentration Test for LA, ALA and GLA

[0082] Level of virus used (MOI of 1) of wild type SARS-CoV-2, WA1/2020 strain in 100 ul of medium (DMEM containing 2% FBS) with indicated concentration of the samples provided samples in a final volume of 100 ul at 37° C. for 10 min in a CO.sub.2 incubator achieving the desired dilution of sample. Control infection samples contained virus (10 ul) alone with 90 ul of medium. Mock well contained 100 ul of medium alone. Vero E6 cells (10,000/well) are plated overnight. Spent medium to be removed and cells were washed once with the complete medium. Fresh complete medium was added to each well (75 ul) along with the sample (25 ul) from Step 6 diluted (1:4 fold as indicated in column I) to each well in triplicate making the final dilutions as indicated in column J. Vero cells were infected for one hour. Subsequently the medium was removed and cells were washed twice with complete medium. Fresh complete medium alone (100 ul) was added to each well and cells were further incubated for 72 hours. The supernatant was removed and assayed for the level of viable virus by RT-PCR assay. The highest serial dilution which achieved inhibition of the test organism was deemed to be the minimum inhibitory concentration (MIC). The following tables (1A-1E) depict the MIC of the test samples with fatty acids as well as with the synthetic detergent Triton X100 and Triolein, a non-fatty acid for comparison.

TABLE-US-00001 TABLE 1A Test organism: SARS-CoV-2 in Vero6 cell line Test sample: Triton X100 (1.71M) Dilution Test 1:1024000 1:512000 1:256000 1:128000 1:64000 1:32000 1 − − − − − +

TABLE-US-00002 TABLE 1B Test organism: SARS-CoV-2 in Vero6 cell line Test sample: Triolein Concentration Test 6.25 mM 12.5 mM 25 mM 50 mM 1 − − − +

TABLE-US-00003 TABLE 1C Test organism: SARS-CoV-2 in Vero6 cell line Test sample: LA(4.32 mM) Dilution Test 1:38400 1:19200 1:9600 1:4800 1:2400 1:1200 1:600 1 − − − − − − +

TABLE-US-00004 TABLE 1D Test organism: SARS-CoV-2 in Vero6 cell line Test sample: ALA (11.78 mM) Dilution Test 1:4800 1:2400 1:1200 1:600 1 − − − +

TABLE-US-00005 TABLE 1E Test organism: SARS-CoV-2 in Vero6 cell line Test sample: GLA (2.96 mM) Dilution Test 1:76800 1:38400 1:19200 1:9600 1:4800 1:2400 1 − − − − + +

Example 2

Antiviral Activities of LA, ALA and GLA

[0083] Antiviral activity of the given composition was evaluated using vero cells and SARS-CoV-2 virus (WA1/2020) strain. First, virus was incubated for 10 minutes with suggested concentration of fatty acids (see table 2A) along with the mock control (virus only). These mixtures were diluted four times to obtain the non-toxic dilution and equal amount of these were added to the seeded vero cells. These infected cells were then incubated for 48/72 hours and subsequently the supernatant was collected. This supernatant was subjected to virus measurement using plaque assay. The results are represented in terms of percentage viral reduction in the sample in Table 2B below. The graphical representation of antiviral activity of LA and GLA is shown in FIG. 1 and FIG. 2 respectively.

TABLE-US-00006 TABLE 2A Conc./dilution used for pre-incubation Conc./dilution Sample with virus added to cells Virus only NA NA Triton X  50 μM  1:12800 Triolein 200 mM 50 mM LA  1:600(4.317 mM) 1:2400 GLA 1:12000(2.9632 mM)  1:4800 ALA 1:300 (11.78 mM) 1:4800

TABLE-US-00007 TABLE 2B Sample % viral reduction Virus only NA (0) Triton X100 95.23 Triolein  2.21 ALA 97.13 LA 97.42 GLA (1:1200) 96.98

[0084] The above results clearly show that the activities of LA, ALA and GLA can be very well compared to the synthetic detergent Triton X. The experiment clearly shows that these fatty acids have the ability to reduce viral load.

Example 3

Compositions Comprising LA, ALA and GLA with their Particle Size

[0085] The present composition was standardized to check for the most stable composition and for their particle size. The control was first run with a blank trial, i.e. without the use of fatty acid. Then various concentrations of the composition were standardized with different percentage of the composition and the batches were prepared using different processes such as magnetic stirring and by HSH of IKA at different centrifugal speed. It was seen that magnetic stirring results in the best of batches for the requisite composition.

[0086] The resultant batches were then checked for various parameters such as: [0087] a) Microscopy [0088] b) Particle size analysis [0089] c) Phase separation

[0090] Following tables depict the compositions of different preparation sample for the preparation of the nasal/mouth spray.

[0091] A) Fatty Acids ALA, GLA and LA for Particle Size and Stability of Composition ALA

[0092] Composition

TABLE-US-00008 TABLE 3A S. No. Ingredients Quantity 1. ALA (ml) 1.5 2. Xantham gum (mg) 10 3. Guar gum (mg) 10 4. Tween 80 (ml) 0.5 5. Span 80 (ml) 0.5 6. Benzalkonium chloride (ml) 0.016 7. Water (ml) 8

[0093] Particle Size Analysis

[0094] Particle size was analyzed using Malvern zetasizer. Results are displayed in the table below and in FIG. 3.

TABLE-US-00009 TABLE 3B S. No. Dilution Mean size pI 1. 50 170.4 0.412

[0095] GLA

[0096] Composition

TABLE-US-00010 TABLE 3C S. No. Ingredients Quantity 1. GLA (ml) 1.5 2. Xanthan gum (mg) 10 3. Guar gum (mg) 10 4. Tween 80 (ml) 0.6 5. Span 80 (ml) 0.6 6. Benzalkonium chloride (ml) 0.016 7. Water (ml) 10

[0097] Particle Size Analysis

[0098] Particle size was analyzed using Malvern zetasizer. Results are displayed in the table below and in FIG. 4.

TABLE-US-00011 TABLE 3D S. no. Dilution Mean size pI 1. 50 283.2 0.565

[0099] LA

[0100] Composition

TABLE-US-00012 TABLE 3E S. No. Ingredients Quantity 1. LA (ml) 1.5 2. Xantham gum (mg) 10 3. Guar gum (mg) 10 4. Tween 80 (ml) 0.6 5. Span 80 (ml) 0.6 6. Benzalkonium chloride (ml) 0.016 7. Water (ml) 8

[0101] Particle Size Analysis

[0102] Particle size was analyzed using Malvern zetasizer. Results are displayed in the table below and in FIG. 5.

TABLE-US-00013 TABLE 3F S. No. Dilution Mean size pI 1. 50 183.0 0.412

[0103] It was observed that the above compositions were stable for more than 100 hours at room temperature

[0104] Cyclodextrin

[0105] Composition

TABLE-US-00014 TABLE 3G S. No. Ingredients Quantity 1. ALA (ml) 1.5 2. Xantham gum (mg) 10 3. Guar gum (mg) 10 4. Cyclodextrin (ml) 0.6 5. Span 80 (ml) 0.6 6. Benzalkonium chloride (ml) 0.016 7. Water (ml) 8

[0106] Many modifications and other embodiments of the invention set forth herein will readily occur to one skilled in the art to which the invention pertain having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the invention is not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims. Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.