Nasal drop composition and device thereof

Abstract

The present invention relates to the use of active substances and compositions comprising them for treating mucosal surfaces to inactivate or kill enveloped viruses. The invention also relates to specific liquid disinfecting compositions and to devices for exerting such antiviral effects. This is achieved, optionally in combination with a vitamin B3 compound, using PPAR activation of fatty acids.

Description

Technical Field

[0001] The present invention relates to the use of active substances and compositions and devices containing them for treating mucosal surfaces to inactivate or kill enveloped viruses. Background Art

[0002] Respiratory tract infections are considered one of the most prevalent causes of illness worldwide. Globally, acute lower respiratory tract infections are a significant cause of morbidity and mortality in children under five years of age. Scientific studies have identified respiratory syncytial virus (RSV) as the most common viral cause of death due to this infection; other major viruses are human metapneumovirus, parainfluenza virus, influenza viruses A and B, adenovirus, and coronaviruses of recent origin. Tens of millions of such infections are reported each year, resulting in more than half a million deaths in children under five years of age, primarily in low- and middle-income families.

[0003] Viruses as described above are known to spread through respiratory aerosol droplets or through contact with infected hands and even through inanimate surfaces. Therefore, hand hygiene is a common method for killing such germs and thus reducing the risk of not only respiratory infections, but also gastrointestinal diseases. Hand hygiene includes washing hands with soap and running water, which not only makes them free of dirt, but also washes off and kills germs including bacteria and viruses. Alternatively, alcohol-based hand sanitizers containing very little water can be applied to the hands as another effective means of removing viruses and bacteria from the hands. Although these methods are very good in terms of instant killing of germs, they are sometimes considered to be caustic. Therefore, there is a need for active substances and compositions that are gentle on mucosal surfaces but effective enough in killing or inactivating viruses to prevent or alleviate infection.

[0004] The inventors, in their search for such actives, discovered a particular type of fatty acid known as a PPAR (peroxisome proliferator activated receptor) activating fatty acid, which has now been found to kill or inactivate enveloped viruses, which was previously unknown. Furthermore, it has been found that it acts synergistically with a particular vitamin, namely vitamin B3 or its precursor or its analog, to provide enhanced antiviral activity. Furthermore, it has been found that vitamin B3 or its precursor or its analog itself can also provide such activity. The PPAR fatty acids according to the present invention include fatty acids having a PPAR effect, and also include their corresponding monoglyceride, diglyceride and triglyceride forms.

[0005] It was therefore an object of the present invention to provide novel active substances which kill or inactivate enveloped viruses.

[0006] It is another object of the present invention to provide this benefit using commonly available active ingredients that are mild to the body's mucosal surfaces. Summary of the invention

[0007] A first aspect of the present invention relates to the use of peroxisome proliferator-activated receptor (PPAR) activating fatty acids for inactivating or killing enveloped viruses on mucosal surfaces of the human or animal body.

[0008] A preferred aspect of the present invention relates to the use of a composition comprising (i) a vitamin B3 compound or a precursor or analog thereof and (ii) a peroxisome proliferator-activated receptor (PPAR) activating fatty acid for inactivating or killing enveloped viruses on mucosal surfaces of the human or animal body.

[0009] Another aspect of the present invention relates to a nasal drop composition comprising:

[0010] (i) 0 to 5.0% by weight of a vitamin B3 compound or an analog thereof,

[0011] (ii) 0.01 to 5.0% by weight of a peroxisome proliferator-activated receptor (PPAR) activating fatty acid; and

[0012] (iii) 0.001 to 5% by weight of a preservative; and

[0013] (iv) 70 to 99.9% by weight of water.

[0014] A further aspect of the present invention relates to a nasal spray device comprising a spray pump capable of spraying the composition from a container containing the nasal drop composition of the present invention. DETAILED DESCRIPTION

[0015] By reading the following detailed description and the appended claims, these and other aspects, features and advantages will become apparent to those of ordinary skill in the art. For the avoidance of doubt, any feature of one aspect of the present invention may be used in any other aspect of the present invention. The word "comprising" is intended to mean "including", but not necessarily "consisting of" or "consisting of". In other words, the steps or options listed do not need to be exhaustive. It should be noted that the embodiments given in the following description are intended to illustrate the present invention and are not intended to limit the present invention to those embodiments themselves. Similarly, all percentages are weight / weight percentages unless otherwise indicated. Except in the operating and comparative examples, or in the case of otherwise clearly indicated, all the numbers representing the amount of material or reaction conditions, the physical properties of the material and / or the use in this specification and claims are modified by the word "about". The numerical range expressed in the format of "x to y" should be understood to include x and y. When multiple preferred ranges are described in the form of "x to y" for a particular feature, it should be understood that all ranges combining different endpoints are also considered.

[0016] The present invention can be used to inactivate or kill viruses, especially enveloped viruses, on mucosal surfaces of human or animal bodies.Therefore, the surface is a living surface.

[0017] Live surfaces include the outer surfaces of living organisms such as plants and animals (including humans). Due to the abundant availability of nutrients and water on most live surfaces, microorganisms reside on these surfaces and can breed on these surfaces. The present invention is used to resist viral infections on mucosal surfaces. Mucosal surfaces according to the present invention include surfaces in the nasal cavity, throat and postnasal cavity.

[0018] The present invention can be used via a leave-on composition. A leave-on composition is a composition that is applied to a mucosal surface of the human body and allowed to remain thereon.

[0019] The present invention relates to the use of PPAR activating fatty acids for inactivating or killing enveloped viruses on mucosal surfaces of the human or animal body. The use is preferably non-therapeutic. The enveloped virus is preferably SARS-Cov2.

[0020] The present invention also enables the prevention of reinfection of mucosal surfaces by inactivating or killing enveloped viruses on these surfaces using one or more of the active substances claimed herein. This means that surfaces treated with such active substances are protected against future infection of the surface within a few hours after such treatment.

[0021] Envelope viruses according to the present invention include coronaviruses or influenza viruses. A preferred coronavirus that can be treated according to the present invention is SARS-Cov2. A preferred influenza virus that can be treated according to the present invention is H1N1. Other envelope viruses that can be treated according to the present invention include RSV (respiratory syncytial virus) and HSV (herpes simplex virus).

[0022] Peroxisome proliferator-activated receptor (abbreviated as PPAR herein) is a transcription factor that controls lipid metabolism. There are three subtypes, PPARα, PPARβ / δ and PPARγ, all of which are located in the skin. A series of specific fatty acids activate these factors, thereby leading to anti-inflammatory effects to reduce skin irritation reactions and pro-differentiation / anti-proliferation reactions, and normalize skin metabolism and provide additional skin care benefits. It is particularly desirable to select PPAR fatty acids containing hydroxyl and / or methyl side chains. Many such acids contain 14 to 30 carbons. Examples of PPAR fatty acids with demonstrated PPAR activating activity are cis-octadecatetraenoic acid, cis-9-trans-11 conjugated linoleic acid, columbinic acid, docosahexaenoic acid, eicosapentaenoic acid, hexadecatrienoic acid, trans-linolenic acid (an isomer of linolenic acid), petroselinic acid, pinolenic acid, punicic acid, ricinoleic acid, trans-ricinolaidic acid (an isomer of ricinoleic acid), stearidonic acid, trans-10-cis-12 conjugated linoleic acid, 7-trans-octadecanoic acid, vaccenic acid, octadecenedioic acid, and hydroxystearic acid.

[0023] The PPAR fatty acids of the present invention also include hydrolyzable PPAR precursors. Potential sources of hydrolyzable PPAR precursors include triglycerides such as coriander seed oil, impatiens seed oil, parinarium laurinarium kernel fat or sabastiana brasilinensis seed oil (for cis-octadecatetraenoic acid) for petroselinic acid, dehydrated castor seed oil for conjugated linoleic acid, and aquilegia oil for octadecatrienoic acid. If a single hydrolyzable precursor of a PPAR activating fatty acid is used, it specifically excludes borage oil, castor oil, and sunflower seed oil. Ideally, the PPAR acid contains 16 or 18 carbon atoms.

[0024] A particularly preferred PPAR acid is hydroxystearic acid or an ester thereof. A most preferred PPAR acid is hydroxystearic acid.

[0025] Preferably, the hydroxystearic acid is 10-hydroxystearic acid, 12-hydroxystearic acid or trihydroxystearic acid (e.g. 9,10,13-trihydroxystearic acid) or trihydroxystearin or a compound that produces one or more hydroxystearic acid or hydroxystearic acid ester molecules when it decomposes, such as a monoester, diester or triester of glycerol and hydroxystearic acid. Among them, 10-hydroxystearic acid, 12-hydroxystearic acid and 9,10,13-trihydroxystearic acid are more preferred, and 12-hydroxystearic acid (12-HSA) is most preferred. 12-HSA has the structure given below:

[0026]

[0027] It is preferred that the PPAR activating fatty acid used in any aspect of the invention is also a caspase 8 modulator.

[0028] According to a preferred aspect, the composition according to the invention may additionally comprise an antimicrobial peptide (AMP). A further preferred aspect relates to the use of a composition according to the invention additionally comprising an antimicrobial peptide. A preferred AMP is LL37.

[0029] A preferred aspect of the present invention relates to the use of a composition comprising (i) a vitamin B3 compound, a precursor thereof or an analog thereof and (ii) a peroxisome proliferator-activated receptor (PPAR) activating fatty acid for inactivating or killing enveloped viruses on mucosal surfaces of the human or animal body. The use is preferably non-therapeutic.

[0030] Yet another aspect of the present invention relates to the use of a PPAR activating fatty acid as an enveloped virus killing or inactivating agent in a composition further comprising a vitamin B3 compound, a precursor thereof, or an analog thereof.

[0031] Yet another aspect relates to the use of a combination of a PPAR activating fatty acid and a vitamin B3 compound, a precursor thereof, or an analog thereof in a mucosal composition as an enveloped virus killing or inactivating system.

[0032] The vitamin B3 compound, its precursor or its analogue is selected from one or more of tryptophan, niacin, nicotinic acid, isonicotinamide, pyridineamide and nicotinamide (also known as niacinamide). Preferred is nicotinamide. Nicotinamide, also known as pyridine-3-carboxamide, is the active, water-soluble form of vitamin B3. According to the present invention, analogues of vitamin B3 also include derivatives, such as cycloalkyl nicotinamide, wherein the cycloalkyl group has 3 to 6 carbon atoms.

[0033] When included in the composition of the present invention, it contains an effective amount of niacinamide, and its concentration is generally 0.05 to 3% by weight of the composition, preferably 0.1 to 2%. In such a composition, the content of hydroxystearic acid is preferably 0.05 to 3% by weight of the composition, more preferably 0.1 to 2%.

[0034] Without wishing to be bound by theory, the inventors believe that the active substances claimed in the present invention, namely PPAR fatty acids, such as 12-HSA, activate host cells to help drive the defense benefit against non-enveloped viruses such as SARS-CoV2. They believe that this mode of action is very different from direct antiviral agents (such as low boiling point alcohols, bleaching agents and cationic surfactants such as quaternary ammonium compounds) that act directly on viral particles to inactivate them, thereby providing antiviral benefits.

[0035] It can be further understood through experiments that active substances (such as PPAR fatty acids) are deposited on the topical surface in sufficient amounts, even from a rinse-off composition. In addition, it is observed that the active substances move into the skin to activate AMP from cells. The production of AMP is a biological process, and this takes a long time. It has been seen that the maximum amount of AMP is produced approximately 48 to 72 hours after the active substance is deposited.

[0036] According to yet another aspect of the present invention, there is provided a composition for the care or cleaning of mucosal surfaces for the inactivation or killing of enveloped viruses on mucosal surfaces of human or animal bodies, the composition comprising (i) a PPAR activating fatty acid and (ii) a mucosal acceptable carrier selected from one or more of water, a wetting agent and an emulsifier. The use is preferably non-therapeutic. Such a composition may further comprise a vitamin B3 compound, a precursor thereof or an analog thereof.

[0037] The present invention also relates to a method for inactivating or killing enveloped viruses on mucosal surfaces of the human or animal body, the method comprising the step of exposing the desired surface to a PPAR activating fatty acid or a composition thereof. The method also includes aspects wherein the composition additionally comprises a vitamin B3 compound, a precursor thereof or an analog thereof. The method according to the present invention is for cosmetic use, i.e. it is for non-therapeutic applications.

[0038] According to a preferred aspect of the present invention, there is provided a nasal drop composition comprising:

[0039] (i) 0.01 to 5.0% by weight of a vitamin B3 compound, a precursor thereof or an analog thereof,

[0040] (ii) 0.01 to 5.0% by weight of a peroxisome proliferator-activated receptor (PPAR) activating fatty acid; and

[0041] (iii) 0.001 to 5% by weight of preservatives; and

[0042] (iv) 70 to 99.9% by weight of water.

[0043] The nasal drop composition of the present invention is usually applied to the nasal cavity using a nasal spray device. Therefore, the present invention also provides a nasal spray device, which includes a spray pump capable of spraying the nasal drop composition of the present invention from a container containing the nasal drop composition. The preservative is preferably benzalkonium chloride (BKC) or benzethonium chloride (BZC), preferably BKC. The nasal drop composition of the present invention may additionally include 0.01 to 10% of a surfactant, which is preferably a nonionic surfactant. The nonionic surfactant that can be used is polyethoxylated fatty acid sorbitan esters, ethoxylated fatty acids, esters of polyethylene glycol, ethoxylates of fatty acid monoglycerides and diglycerides, and ethylene oxide / propylene oxide block polymers.

[0044] The invention will now be illustrated by the following non-limiting examples.

[0045] Example

[0046] Example A, 1-3: Effect of 12-HSA on virus killing on the skin keratinocyte cell line HaCaT

[0047] According to the standard protocol, the skin keratinocyte cell line HaCaT was differentiated, then treated with 10 μM, 20 μM and 40 μM 12HSA, and the treatment was carried out for 72 hours. After this treatment, the secretion group containing secreted antimicrobial peptides (AMPs) was collected and incubated with the SARS-CoV2 virus for 4 hours. After incubation for 4 hours, the virus killing was counted by using a quantitative PCR-based method. The data of viral gene expression as a percentage of the control are shown in Table-1 below:

[0048] Table-1

[0049]

[0050] The data in the above table indicate that the secretome produced in the presence of 12HSA is capable of killing / inactivating viruses.

[0051] Examples BD, 4-6: Effects of 12-HSA and Vitamin B3 and Combinations on Epithelial Cells (Calu-3 Cells):

[0052] Compound treatment and plaque formation assay

[0053] 0.1MOI SARS-CoV-2 was adsorbed on Calu-3 cells and seeded at approximately 120,000 cells / well in a 48-well plate for 1 hour. After washing, fresh medium was added along with nicotinamide (16.4mM), 12HSA (10μM) and their corresponding vehicle controls. After 48 hours, the conditioned medium (containing viral particles) was collected and the plaque forming units (pfu / ml) of each treatment were determined by plaque formation assay on Vero-E6 cells (48-well plate).

[0054] The data are summarized in Table 2 below:

[0055] Table-2

[0056]

[0057] The data in the table above demonstrate that both 12-HSA and nicotinamide are able to inhibit SARS-CoV-2 as demonstrated by the reduction in plaque forming units from the assay used above. The combination of 12-HSA and vitamin B3 significantly improved inhibition.

[0058] Examples EG, 7-9: Effect of human keratinocyte supernatant on SARS-CoV2 infectivity :

[0059] Differentiated primary human keratinocytes were incubated in serum-free Epilife medium containing the concentrations of nicotinamide and / or 12HSA for 72 hours, and the conditioned medium was collected. SARS-CoV-2 virus particles were incubated with the above conditioned medium for about 4 hours. The pfu / ml of these treated virus particles was determined by plaque or TCID50 using VeroE6 reporter cells. The % infection relative to the control is shown in Table-3 below:

[0060] Table-3

[0061]

[0062]

[0063] The data in the above table indicate that similar effects as those in Table 2 can be obtained using primary human keratinocytes.

[0064] Example HN: Effects of some vitamin B3 compounds, their precursors or their analogs on skin keratinocyte cell lines Impact of virus killing

[0065] According to standard protocol, skin keratinocyte cell line CRL4048 is differentiated, then treated with the vitamin B3 compound of various concentrations as shown in Table 4 below, and the treatment is carried out for 72 hours. After this treatment, the secretion group containing secreted antimicrobial peptides (AMP) is collected, and incubated 4 hours with SARS-CoV2 virus. After incubation for 4 hours, by incubating virus mixture on VeroE6 reporter cell line and using the method based on quantitative PCR for measuring infection to count virus killing. The data of viral gene expression are presented in Table 4 below as the percentage of control:

[0066] Table-4

[0067]

[0068] The data in Table 4, together with the data in Table 1 above, indicate that 12HSA is far superior to vitamin B3 compounds, as nicotinamide (Examples I and J) is ineffective at similar (micromolar) concentrations. When other vitamin B3 compounds (Examples KN) were used, orders of magnitude higher (millimolar rather than micromolar) concentrations were required to kill the virus compared to 12HSA (Examples 1-3 of Table 1).

[0069] Examples P, 10-14: Effects of other PPAR fatty acids on viral killing on a skin keratinocyte cell line

[0070] Other PPAR fatty acids were used to study viral gene expression as shown in Table 5 below. The procedure used to generate the data was similar to that used for Table 4. The data are summarized in Table 5 below:

[0071] Table-5

[0072]

[0073] In the table above, CLA stands for conjugated linoleic acid.

[0074] The data in Table 5 above demonstrate that other PPAR fatty acids (Examples 10-14) are as effective as 12HSA (Examples 1-3 in Table 1) in viral gene expression at similar (micromolar) concentrations.

Claims

1. Use of peroxisome proliferator-activated receptor (PPAR) activating fatty acids for inactivating or killing enveloped viruses on mucosal surfaces of the human or animal body.

2. Use of a composition comprising (i) a vitamin B3 compound, a precursor thereof or an analog thereof and (ii) a peroxisome proliferator-activated receptor (PPAR) activating fatty acid for inactivating or killing enveloped viruses on mucosal surfaces of the human or animal body.

3. The use according to claim 1 or 2, wherein the enveloped virus is a coronavirus, preferably SARS-CoV2; or an influenza virus.

4. Use according to any one of the preceding claims, wherein the PPAR activating fatty acid is hydroxystearic acid (HAS), preferably 12-hydroxystearic acid or 10-hydroxystearic acid.

5. The use according to any one of the preceding claims, wherein the vitamin B3 compound, its precursor or its analogue is selected from one or more of tryptophan, nicotinic acid, niacin, isonicotinamide, pyridineamide and nicotinamide.

6. Use of a composition for caring for or cleaning mucosal surfaces for inactivating or killing enveloped viruses on mucosal surfaces of human or animal bodies, the composition comprising (i) a PPAR-activating fatty acid and (ii) a mucosal-acceptable carrier selected from one or more of water, a wetting agent and an emulsifier.

7. Use of a composition according to claim 6, which further comprises a vitamin B3 compound, a precursor thereof or an analogue thereof.

8. A method of inactivating or killing enveloped viruses on a mucosal surface of a human or animal body, the method comprising the step of exposing the desired mucosal surface to a PPAR activating fatty acid or a composition thereof.

9. The method of claim 8, wherein the composition further comprises a vitamin B3 compound, a precursor thereof, or an analog thereof.

10. A nasal drop composition comprising (i) 0 to 5.0% by weight of a vitamin B3 compound, a precursor thereof or an analog thereof, (ii) 0.01 to 5.0% by weight of a peroxisome proliferator-activated receptor (PPAR) activating fatty acid; and (iii) 0.001 to 5% by weight of a preservative; and (iv) 70 to 99.9% by weight of water.

11. A nasal spray device comprising a spray pump capable of spraying the composition as claimed in claim 10 from a container containing the composition.

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