Technical Field

The invention relates to an antiviral nasal spray for use in the healthcare sector and other pharmaceutical products that will be developed with the same active ingredients therefrom.

The invention particularly relates to a novel antiviral nasal spray that provides a high level of suppression against the virus by means of the active ingredient content of dill oil and tannic acid.

State of the Art

During the COVID-19 pandemic, millions of people were affected by this viral infection and hundreds of thousands of people lost their lives. As of October 2022, the total number of people diagnosed with Covid-19 was 16.9 million in Turkey and exceeded 623 million worldwide. In this period covering about 3 years, the number of cases that can be detected related to the number of deaths caused by COVID is around 101000 in Turkey and 65000000 worldwide. The incidence of pandemics has increased in the 20th and 21st century, along with outbreaks of SARS and MERS, which are of coronavirus origin like COVID-19, epidemics such as avian flu and swine flu, which are of influenza virus origin, lead to a significant level of threat and loss of life. In an important part of these diseases, the respiratory system (nasal tract) comes to the fore as a transmission route. Viruses that are the causative agents of the disease, after entering the body through the respiratory air, mainly cling to the nasal mucosa and initiate infection. The virus that multiplies here then either goes to the lungs and causes pneumonia, or spreads through the blood to the body, leading to a systemic (general) disease situation. When it comes to these advanced stages where the signs of the disease appear, the treatment options and chances of success for viral infections decrease significantly. Therefore, the most appropriate way to prevent or reduce the damage caused by such infections is to stop the virus at the point of entry into the body, which is still the initial stage of infection, that is, in the nasal mucosa.

The present practices followed for protection from viral infections transmitted through respiration and some of their deficiencies are given below:

The documents found in the patent and literature research conducted for the state of the art are summarized below.

US2019166903 A1 describes an apparatus for vaporizable liquid composition developed for personal use. However, there is no explanation for the combined use of dill oil and tannic acid in the said application. WO2021206688 A1 relates to a compound developed for aromatherapy. However, there is no explanation for the combined use of dill oil and tannic acid in the said application.

Therefore, due to the disadvantages described above and due to the inadequacy of existing solutions on the subject, it became necessary to make an improvement in the relevant technical field.

Brief Description of the Invention

The present invention relates to an antiviral nasal spray that meets the requirements mentioned above, eliminates all the disadvantages, and provides some additional advantages.

The invention is inspired by the present situations and aims to solve the disadvantages mentioned above.

The main object of the invention is to develop a new formulation of antiviral nasal spray. The formulation, which is the subject of the invention, comprises natural ingredients that inactivate the virus and prevent it from multiplying without having a harmful effect on the cells. The main technical advantages of the antiviral nasal spray formulation are given below.

• By means of its active substance composition, it directly targets the virus and stops or reduces its reproduction. It has been shown in experimental laboratory studies that it has this property.

• It comprises completely natural active ingredients, for this reason, there is no risk of side and undesirable effects. The active ingredients used in its composition are obtained from products that are widely consumed as food. These products are consumed by millions of people every day. In addition, cytotoxicity experiments of the active substances contained in the formula have been performed and harmless doses have been determined in laboratory studies.

• Since it is applied in the form of a nasal spray, it stops the infection before it even starts or ends it by blocking it while it is still at the initial stage. In experiments conducted with the infective (live) coronavirus, the active ingredients in its formulation were found to block the virus both in its free state and to stop its entry into the cell and reproduction in the cell.

• It is prepared with easily available raw materials at a much lower cost compared to drug treatments. The dill oil and tannic acid substances contained in the formulation are easily available products and are differed from other active pharmaceutical ingredients in terms of price and logistics. There are no restrictions on national and international trade.

• By means of the supportive substances contained in the formulation, it supports antiviral activity for the following reasons: Polyvinyl alcohol: Allows the active substance components to remain in the nasal mucosa for a longer time and effectively.

> Benzalkonium chloride: It has an anti- inflammation and decongestant effect.

> Phosphate buffer solution (pH 6.00): It is carriage substance of the formulation and increases the antimicrobial activity in the nasal mucosa by means of its adjusted pH.

The structural and characteristic features of the invention and all its advantages will be understood more clearly by means of the figures given below the detailed description written by making references to these figures, and therefore evaluation should be made taking into account these figures and detailed description.

Figures to Help Understand the Invention

In Figure 1 , effectiveness of dill oil on coronavirus is given versus time.

In Figure 2, effectiveness of tannic acid on coronavirus is given versus time.

In Figure 3, effectiveness of dill oil and tannic acid (together) on coronavirus is given versus time.

In Figure 4, the effect of dill oil and tannic acid with active ingredient supplementation is given on the reproduction of coronavirus at 48- hour intervals.

In Figure 5, cell adsorption and penetration rates are given.

In Figure 6, the effect of dill oil and tannic acid (together) is given on inactivating the coronavirus in the medium.

Detailed Description of the Invention

In this detailed description, the preferred embodiments of the invention are explained only for a better understanding of the subject.

The invention relates to an antiviral nasal spray for use in the healthcare sector. The formulation of the antiviral nasal spray contains dill oil and tannic acid as active ingredients, polyvinyl alcohol, benzalkonium chloride, phosphate buffer solution as auxiliary substances. The pH of the phosphate buffer solution is preferably 6. In the invention, dill oil and tannic acid are mandatory ingredients of antiviral nasal spray. Glycerin can be used instead of polyvinyl alcohol. Similarly, physiological saline can be used instead of phosphate buffer solution.

In a preferred application of the invention, the ingredients contained in the antiviral nasal spray formulation content, the intervals of use and the preferred amounts of use are given in the following table.

Dill oil is an herbal extract product comprising fatty acids with antiviral, antimicrobial and antioxidant activity. In experimental applications with coronavirus, it has been determined that it inactivates viruses in the medium at a rate of more than 98% for 4 hours, suppresses the entry of the virus into cells at a rate of more than 90%, and its reproduction in the cell at a rate of 99.58%.

Tannic acid is a compound with antiviral and antimicrobial activity and is found naturally in various plants. In experimental applications with coronavirus, it has been determined that it inactivates viruses in the medium at a rate of more than 98.2% for 4 hours, suppresses the entry of the virus into cells at a rate of more than 58%, and its reproduction in the cell at a rate of up to 100%.

In the studies carried out within the scope of the invention, it was determined that dill oil and tannic acid have a synergetic effect when used together and inactivate the coronavirus in the medium by 99.4%; suppress the entry of the virus into cells by more than 99.58% and the reproduction of the virus in the cell by up to 99.94%. For this reason, it has been evaluated that it would be appropriate to formulate both active substances together in the determined amounts.

Polyvinyl alcohol, by means of its structural property, allows the active substance components to remain in the nasal mucosa for a longer time and effectively.

Benzalkonium chloride provides a supportive effect in reducing mucosa damage due to virus infection by means of its anti-inflammatory and decongestant effect.

Phosphate buffer solution (pH 6.00) essentially acts as a vehicle substance in the formulation, but it supports the physiological defense barrier of the nasal mucosa by means of its balanced pH value. The pH level has been selected as the level with the highest antimicrobial activity in the nasal mucosa. Antiviral effectiveness may decrease if this value is changed upwards.

The antiviral nasal spray, which is the subject of the invention, has been prepared for use in order to prevent the onset or progression of respiratory viral infections. Non-sick individuals can use it to protect themselves against the virus, while sick individuals can use it to slow the progression of the disease and reduce contagion. After emptying the contents in the nose, if any, spray is applied to both nasal cavities 1-2 times. This application should be performed 4 times a day at intervals of six hours. Due to its natural content, it can also be applied at shorter intervals when needed.

In the production method of antiviral nasal spray, which is the subject of the invention, firstly, phosphate buffer solution (PBS) is prepared. PBS is prepared without calcium, and it is sterilized. Preferably, it is started to be spinned in a magnetic mixer in a glass container. Dill oil (preferably at a rate of 0.006-0.015% (v/v)) is added to the prepared solution, followed by Tannic acid (preferably at a rate of 30-70 pM). The added T is in powder form. The next added ingredient is polyvinyl alcohol (preferably 7-14 mg/ml). In the next processing step, benzalkonium chloride (preferably 0.01-0.04mg/ml) is added. The resulting final product is mixed for 30 - 60 minutes at a speed of 200 rpm, preferably at room temperature. Then, it is passed through preferably 0.2 pm membrane filter and sterile filtration process is applied. The final processing step is the filling process. In this process, glass or plastic spray light-proof bottles are filled. In this method, mixing is performed at a temperature of 21-24 ID for 30-60 minutes, sterile filtration is performed at a temperature of 21-24 ID, storage is performed at a temperature of 21-24 ‘C.

The studies conducted to prove that the invention provides an unexpected / surprising effect are summarized below. These studies are aimed at providing information only and they do not have any restrictive meaning.

Abbreviations:

Dill oil : An herbal essential oil

Tannic acid : A powdered compound derived from plants

BCoV : Coronavirus used in the study (Bovine coronavirus)

The Effect of Dill Oil on Coronavirus Reproduction

Virus titers obtained in the treatment of dill oil (live virus numbers) Virus inhibition rates obtained by dill oil treatment (%) (virus inactivation rate)

The effectiveness of dill oil on coronavirus versus time is given in Figure 1. The effective concentration of dill oil was confirmed by this conducted experiment.

The Effect of Tannic Acid on Coronavirus Reproduction Virus titers obtained in the treatment of tannic acid (live virus numbers)

Virus inhibition rates obtained by tannic acid treatment (%) (virus inactivation rate)

The effectiveness of tannic acid on coronavirus versus time is given in Figure 2. The effective concentration of tannic acid was confirmed by this conducted experiment.

The Effect of Dill oil and Tannic Acid (Together) on Coronavirus Reproduction Virus titers obtained in the treatment of dill oil and tannic acid (together) (live virus numbers) Virus inhibition rates obtained dill oil and tannic acid (together) treatment (%) (virus inactivation rate)

Figure 3 shows the effectiveness of dill oil and tannic acid (together) on coronavirus versus time. With this experiment, dill oil and tannic were combined and tested together at the concentrations determined previously to be the most effective. Thus, the synergetic effect was examined. Thus, inhibition was achieved at a rate of 99.94% at 48th hour. It was determined that this effect lasted as >99% for 72 hours and decreased to 97% at 72nd hour.

The Effect of Dill oil and Tannic Acid Together on Coronavirus Reproduction and with Active Ingredient Supplementation at 48-Hour Intervals Virus titers obtained in the treatment of dill oil and tannic acid (together) (live virus numbers)

Virus inhibition rates obtained dill oil and tannic acid (together) treatment (%) (virus inactivation rate) Figure 4 shows the effect of dill oil and tannic acid on coronavirus reproduction together and with active ingredient supplementation at 48-hour intervals. With this experiment, the dill oil and tannic acid at the concentrations that were previously determined to be the most effective are combined its use with supplementing it at intervals of 48 hours has been tested. Thus, the maximum frequency of use (time interval) was evaluated. With this application, the activity that was 99.44% in the 24th hour could be extended until the 120th hour.

The Effect of Dill oil and Tannic acid (together) on the Penetration of Coronavirus

Effect on Virus Titers:

Inhibition rates: Figure 5 shows the adsorption and penetration rates. In this experiment, the potential of dill oil and tannic acid to prevent the penetration of coronavirus into the cell in the medium was tested when they are used separately and together. According to the data obtained, while the adsorption was stopped by 68.38% when the T substance was used alone, it achieved 90% success when the D substance was used alone. However, when the two components are used together, this ratio increases to 98.58%. This is one of the most prominent unexpected findings of this study.

The Effect of Dill oil and Tannic Acid (Together) on the Inactivation of Coronavirus in the Medium

Figure 6 shows the effect of dill oil and tannic acid (together) on inactivating the coronavirus present in the medium. In this experiment, the potential of inactivating the coronavirus present in the medium was tested when dill oil and tannic acid are used separately and together. According to the data obtained, when tannic acid is used alone it inactivates the coronavirus at a rate of 99.43% in the 1st hour, the inactivation rate decreases to 90% during the 2nd hour; when dill oil is used alone the inactivation rate, which is 94.37% in the 1st hour, decreases to 94.37% during the 2nd hour. However, when the two components are used together, this rate starts at 90% in the 1 st hour and continues high until the 4th hour (98.22%). Therefore, if the two components are used together, it can successfully inactivate the viruses present in the medium.