DESC:
Field of the invention
The present invention relates to personal care formulations. Most specifically the present invention relates to an oral care formulation comprising protoberberine.

Background and the prior art
During present times, there are lots of challenges faced by the consumers when it comes to oral care. As the oral care products available in the market are either incapable or involving extensively longer time to provide the expected benefits to the consumers; there are attempts made to offer novel formulations for oral care. For e.g. EP1941865A1 issued to Henkel, related to cosmetic composition, discloses the names of Tinospora cordifolia, thyme oil, basil oil and malic acid.
There are literature available to show that Tinospora is used as antiviral agents (Antiviral prospective of Tinospora cordifolia on HSV-1, Int.J.Curr.Microbiol.App.Sci (2018) 7(1): 3617-3624, Tinospora cordifolia: One plant, many roles, Anc Sci Life. 2012 Apr-Jun; 31(4): 151–159, doi: 10.4103/0257-7941.107344: 10.4103/0257-7941.107344). US20090252796 also discloses the names of Tinospora, thyme oil, malic acid etc.
Basil oil and Thyme oil Essential Oils as Antiviral Agents, Potential of Essential Oils to Treat SARS-CoV-2 Infection: An In-Silico Investigation, Int. J. Mol. Sci. 2020, 21, 3426; doi:10.3390/ijms21103426 discloses essential oil components which may act synergistically, essential oils may potentiate other antiviral agents, or they may provide some relief of COVID-19 symptoms.
However, none of the prior arts discloses the formulation containing the functional actives, which is effective against the pathogens in the oral cavity with 99.99% of efficacy within 60 sec time interval.

Object of the invention
It is an object of the present invention to provide a novel oral care formulation that helps to act against the pathogens in oral cavity within 60 sec.
It is another object of the present invention to provide a novel oral care formulation that is effective against a wide range of pathogens including bacteria, viruses and fungi.
It is yet another object of the present invention to provide a formulation that can be easily made into various product formats including mouthwash, mouth spray, nasal spray, throat spray and the like.

Summary of the invention
Accordingly, the present invention provides an oral care formulation comprising a combination of at least one protoberberine, thyme oil and basil oil, said formulation being effective against pathogens within 60 seconds in the oral cavity.

Detailed description of the present invention
The present invention provides a novel oral care formulation comprising protoberberine. Protoberberine was found to be one of the actives, among the numerous compounds which are present in the plant, Tinospora cordifolia. However, berberine chloride, thyme oil, and basil oil used in the instant invention is obtained from commercially available sources such as from Sigma-Aldrich.
The protoberberines named berberine is an important alkaloid (molecular formula C20H19NO5 and molecular weight of 353.36) identified as one of the important active molecules with antimicrobial activity.
However, the said active, berberine per se shall not be as effective as the formulation of the present invention.
The present inventors have surprisingly found that a combination of berberine chloride, thyme oil, basil oil in specific amounts is effective against a wide range of pathogens and works within 60 seconds in the oral cavity.
Accordingly, the present invention provides an oral care formulation comprising berberine chloride, thyme oil and basil oil. The formulation of the present invention comprises 0.005% to 0.045% by weight; preferably 0.005% to 0.015%; most preferably 0.015% by weight of berberine chloride.
The formulation of the present invention comprises 0.1% to 0.4% by weight; preferably 0.1% to 0.2%; most preferably 0.1% by weight of thyme oil.
The formulation of the present invention comprises 0.05% to 0.2% by weight; preferably 0.05% to 0.1%; most preferably 0.05% by weight of basil oil.
The present inventors have further found that inclusion of malic acid in the oral formulation of the present invention further enhances its anti-pathogenic activity. Malic acid is present in amounts 0.5% to 1.5% by weight; preferably 0.5% to 1.0%; most preferably 0.5% by weight.
The oral formulation of the present invention may further comprise 0.5% to 1.5% by weight malic acid, 0.5% to 1.0% by weight Sodium benzoate, 3% to 5% by weight Propylene glycol and 20% to 25% by weight Isopropanol; preferably 0.5% by weight Malic acid, 0.5% by weight Sodium benzoate, 3% by weight Propylene glycol and 20% by weight Isopropanol.
The actives used in the present invention have been commercially procured.
According to a preferred embodiment of the present invention the oral care formulation of the present invention comprises a combination of Berberine chloride, thyme oil and basil oil. The composition may further comprise of sodium benzoate.
According to another embodiment of the present invention the oral care formulation of the present invention comprises a combination of Berberine chloride, thyme oil, basil oil and malic acid. The composition may further comprise of sodium benzoate.
According to yet another preferred embodiment of the present invention there is provided an oral care formulation comprising 0.015% by weight Berberine chloride, 0.1% by weight thyme oil, 0.05% by weight basil oil and 0.5% by weight sodium benzoate.
According to a further preferred embodiment of the present invention there is provided an oral care formulation comprising a combination of 0.015% by weight Berberine chloride, 0.1% by weight thyme oil, 0.05% by weight basil oil, 0.5% by weight malic acid and 0.5% by weight sodium benzoate.
The oral care formulation of the present invention is effective against a wide range of pathogens including bacteria, viruses and fungi. The bacteria against which the present formulation is effective are Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Enterococcus hirae.
The oral care formulation of the present invention can be formulated into various formats namely mouthwash, mouth-spray, nasal spray, throat spray, and the like.
The present invention is now illustrated by way of non-limiting examples.

Example 1
Table 1 below shows an illustrative example of a Mouthwash formulation in accordance with the present invention
Table 1
Mouthwash complete formulation ingredients %
Berberine chloride 0.015
Thyme oil 0.1
Basil oil 0.05
Wintergreen flavor 0.8
Menthol flavor 0.3
Propylene Glycol 3
Sucralose 0.05
Xylitol 5
Sodium Benzoate 0.5
Malic acid 0.5
NaCl 0.2
Surfectant (Pluacare 127) 1.3
Solubalizer 2 (Cremophor RH 40) 1
Isopropanol 20

Method of preparation
1. Preparation of Surfactant solution (Solution A)
Pluracare 127 (obtained from BASF) at 1.3% concentration is slowly added to 30 ml water and mixed at 300 rpm with the help of a magnetic stirrer until a clear solution is obtained.
2. Solubilization of Essential oils, Flavors and Polyethylene glycol (Solution B)
Cremophor RH 40 (obtained from BASF) at 0.65%, concentration is used as a solubilizer for essential oils and flavors. 0.1% Thyme oil, 0.05% Basil oil, 0.8% Wintergreen, 0.3% menthol flavor and 3% Propylene glycol added sequentially to the solubilizer and mixed well till all the ingredients mix well and a clear solution is obtained. To this clear solution, 20% Isopropanol is added by continuous stirring.
3. Mixing of Surfactant solution with Essential oils and Flavors (Solution C)
Solution A and Solution B are mixed slowly until a clear solution C is obtained.
Addition of other formulation ingredients
To Solution C other formulation ingredients like 0.05% Sucralose, 5% Xylitol, 0.5% Malic acid and 0.2% NaCl are added sequentially until a clear solution is obtained.
4. Addition of Berberine chloride
0.015% Berberine chloride was dissolved in 3% Polyethylene glycol is added finally and mixed.
5. Water is added to make up to the desired volume.

Example 2
Various experiments have been performed to demonstrate the effectiveness of the active components individually and in different combinations as mentioned below. The composition comprises a judiciously chosen active and weight % to act against various pathogens within 60 sec time-frame.

Log kill explanation - As an example, if there are one million microbes present on a surface, then
1 log reduction would reduce the number of microbes by 90 percent
2 log reduction removes 99 percent
3 log reduction removes 99.9 percent
4 log reduction removes 99.99 percent
5 log reduction removes 99.999 percent

Non-working example
Experimental details – Anti bacterial efficacy tested as per EN 1276 protocol
Test organism – Staphylococcus aureus
Contact time – 1 min
Soiling condition – 0.3% BSA
Temperature – 20ºC

Table 2
Sample # Individual actives
Actives Log kill Formulation constituents
1 0.015% Berberine chloride 0.3 0.015% Berberine chloride solution prepared in 3% polyethylene glycol
2 0.1% Thyme oil 1.23 0.1% Thyme oil solution prepared in water
3 0.05% Basil oil 0.5 0.05% Basil oil solution prepared in water
4 0.5% Malic acid 0 (No activity) 0.5% Malic acid solution prepared in water

The data in Table 2 demonstrates that the actives (sample no 1-4) when tested individually exhibited very less or no rapid bactericidal activity. The log kill observed against S.aureus after contact with each natural for 1 min was less than 2.0.

Table 3
Active Concentration range Log kill Formulation constituents

Berberine chloride 0.045% 0.27 0.045%, 0.015% and 0.005% Berberine chloride prepared in 3% Polyethylene glycol
0.015% 0.3
0.0050% 0.08

Thyme oil 0.4% 0.19 0.4%, 0.2% and 0.1% Thyme oil solutions prepared in water
0.2% 0.35
0.1% 1.23

Basil oil 0.20% 4.4 0.2%, 0.1% and 0.05% Basil oil solutions prepared in water
0.10% 0.6
0.05% 0.5

Addition of 0.2% Basil oil gives a very intense off flavour to the formulation, which was not acceptable by the consumer.
The above data in Table 3 demonstrates that when the actives were tested individually at 1X, 2X, 4X concentrations they exhibited very less or no rapid bactericidal activity. Thus, conferring that individual active at higher concentrations except for 0.2% Basil oil do not exhibit bactericidal activity more than 2.0 log.

Table 4
Sample # Formulation ingredients
Actives Log kill Formulation constituents
4 0.5% Sodium Benzoate 0 0.5% Sodium benzoate solution prepared in water
5 0.5% Malic Acid 0 0.5% Malic acid solution prepared in water
6 20% Isopropanol 0 20% Isopropanol solution prepared in water

The above data in Table 4 demonstrates that the formulation ingredients (sample no. 4-6) exhibited no rapid bactericidal activity.

Optimum/base (less preferred) data range
Table 5
3 Actives + Formulation ingredients
Sample # Actives Log kill
Formulation constituents
7 0.045 % Berberine chloride + 0.3 % Thyme oil + 0.15% Basil oil + 0.5% Malic acid + 0.5% Sodium Benzoate 2.0 0.045% Berberine chloride prepared in 3% Polyethylene glycol and to this , 0.3% Thyme oil, 0.15% Basil oil, 0.5% Malic acid and 0.5% Sodium Benzoate were added
8 0.005 % Berberine chloride + 0.03 % Thyme oil + 0.016 % Basil oil 3.5 0. 005% Berberine chloride was prepared in 3% Propylene glycol and to this , 0.03% Thyme oil, 0.016 % Basil oil, 0.5% Malic acid and 0.5% Sodium Benzoate were added

The above data in Table 5 demonstrates that when the combination of three actives (sample no. 7-8) tested at ranges 3X times higher and 3X lower concentrations than the preferred combination exhibited less rapid bactericidal activity.

Best working example (the most preferred)
Table 6
3 Actives + Formulation ingredients
Sample # Actives Log kill
Formulation constituents
9 0.015% Berberine chloride+ 0.1% Thyme oil + 0.05% Basil oil +0.5% Malic acid + 0.5% Sodium Benzoate >5.0 0.015% Berberine chloride, 0.1% Thyme oil, 0.05% Basil oil, 0.5% Malic acid and 0.5% Sodium benzoate solution prepared in water

The above data in Table 6 demonstrates that this specific combination of three ingredients along with formulation ingredients (Sample no 9) has bactericidal activity of >5.0 log. Thus, conferring the above combination has worked synergistically and have shown significant bactericidal activity of >3.0 log which corresponds to 99.99% protection against S. aureus.

Sample #9 was tested for antimicrobial efficacy against other bacteria
Table 7
Sample no Organism Log kill
#9 Escherichia coli >5
#9 Pseudomonas aeruginosa >5
#9 Enterococcus hirae >5

Generally, more than 3 log kill is the primary requisite for any oral care, sanitizers and disinfectants. The present active system exhibits > 5.0 log activity against S. aureus, E.coli, P.aeruginosa and E.hirae.
,CLAIMS:
1. An oral care formulation comprising a combination of at least one protoberberine, thyme oil and basil oil, said formulation being effective against pathogens within 60 seconds in the oral cavity.

2. The oral care formulation as claimed in claim 1, wherein said protoberberine is Berberine chloride.

3. The oral care formulation as claimed in claim 1, wherein said formulation further comprises malic acid.

4. The oral care formulation as claimed in any of the preceding claims, wherein said combination of berberine chloride, thyme and basil oil are present in amounts preferably, 0.045%, 0.3% and 0.15% by weight, with 99% efficacy (log kill value of 2); more preferably, the said combination of berberine chloride, thyme and basil oil is present in amounts of 0.005%, 0.03% and 0.016% by weight, with efficacy of 99.94% (log kill value of 3.5) and most preferably, the said combination of berberine chloride, thyme and basil oil is present in amounts of 0.015%, 0.1% and 0.05% by weight with efficacy of >99.999% (log kill value of 5).

5. The oral care formulation as claimed in any of the preceding claims, wherein said thyme oil is present in amounts 0.1% to 0.4% by weight; preferably 0.1% to 0.2%; most preferably 0.1% by weight.

6. The oral care formulation as claimed in any of the preceding claims, wherein said basil oil is present in amounts 0.05% to 0.2% by weight; preferably 0.05% to 0.1%; most preferably 0.05% by weight.

7. The oral care formulation as claimed in claim 3, wherein malic acid is present in amounts to 0.5% to 2.0% by weight; preferably 0.5% to 1.0%; most preferably 0.5% by weight.

8. The oral care formulation as claimed in any of the preceding claims, wherein said formulation further comprise of 0.5% by weight Malic acid, 0.5% by weight Sodium benzoate, 3% by weight Propylene glycol and 20% by weight Isopropanol.

9. The oral care formulation as claimed in any of the preceding claims, wherein said pathogens include bacteria, viruses and fungi.

10. The oral care formulation as claimed in claim 9, wherein said bacteria is selected from Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Enterococcus hirae.

11. An oral care formulation comprising 0.015% by weight Berberine chloride, 0.1% by weight thyme oil, 0.05% by weight basil oil and 0.5% by weight sodium benzoate.

12. An oral care formulation comprising a combination of 0.015% by weight Berberine chloride, 0.1% by weight thyme oil, 0.05% by weight basil oil, 0.5% by weight malic acid and 0.5% by weight sodium benzoate.

13. The oral care formulation as claimed in any of the preceding claims, wherein said formulation is formulated into various formats namely mouthwash, mouth-spray, nasal spray, throat spray, and the like.