Combinations for oral compositions, their preparation and use

Abstract

A composition having a pH in the range of from 3 to 8.5, comprising: (a) in the range of from 0.1% to <10% w/w (based on the total weight of the composition) of a stock solution comprising a mixture of bioflavonoids and fruit acids or salts thereof; (b) sodium hyaluronate; and (c) water; and, optionally, (d) a pharmaceutically acceptable carrier therefor; wherein the sodium hyaluronate has an average molecular weight of between 800,000 and 4,000,000.

Claims

1. An oral care composition having a pH of 3 to 8.5 comprising: (a) from 0.1 to 10% w/w based on the total weight of the composition, of a stock solution comprising (i) 0.45% to 9% of an extract or biomass comprising a mixture of water soluble bioflavonoids derived from the pith of immature bitter orange wherein naringin and neohesperidin comprise in excess of 75% of the mixture of bioflavonoids and (ii) citric acid; (b) sodium hyaluronate of an average molecular weight between 800,000 and 4,000,000 daltons; (c) water; and (d) optionally a pharmaceutically acceptable carrier therefore.

2. A composition according to claim 1, wherein the molecular weight of the sodium hyaluronate is between 1,000,000 and 2,000,000 daltons.

3. A composition according to claim 2, comprising 0.2 to 10% w/w of sodium hyaluronate.

4. A composition according to claim 3, comprising 0.2 to 1% w/w of sodium hyaluronate.

5. A composition according to claim 2, comprising 0.005 to 0.1% w/w of sodium hyaluronate.

6. A composition according to claim 5, comprising 0.01% w/w of sodium hyaluronate.

7. A composition according to claim 1, which comprises in the range of from 0.1 to 5% w/w of the stock solution.

8. A composition according to claim 1, wherein the stock solution has a pH in the range of from 3.5 to 8.

9. A composition according to claim 8, wherein the composition has a pH in the range of from 4 to 7.

10. A composition according to claim 9, wherein the composition has a pH in the range of from 5 to 6.5.

11. A composition according to claim 1, wherein the bioflavonoid mixture comprises from 40 to 60% w/w biomass, based on the weight of the bioflavonoid mixture.

12. A composition according to claim 1, comprising in the range of from 0 to 30% w/w alcohol, based on the weight of the composition.

13. A composition according to claim 12, wherein the alcohol is ethanol.

14. A composition according to claim 1, which is free of alcohol.

15. A composition according to claim 1, having a pH in the range of from 5 to 6.5, comprising 20 to 80% w/w water and from 0 to 30% w/w alcohol; and, optionally a pharmaceutically acceptable carrier therefor.

16. A composition according to claim 1, wherein the component (d) is selected from fluoridating agents, flavours, sweeteners, colours and preservatives; anti-plaque and antimicrobial agents; humectants, surfactants, thickeners, gums, binders and abrasive agents; and film-formers.

17. A composition according to claim 1 which is a toothpaste containing 20 to 40% w/w water or a liquid composition comprising 60 to 80% water.

18. A composition according to claim 1, which is in the form of a paste, cream, ointment, gel or liquid.

19. An oral composition according to claim 18, which is in the form of toothpaste, mouthspray, mouthrinse or mouthwash.

20. A method for preparing a composition as claimed in claim 1, comprising bringing components (a), (b), (c) and optionally (d) into intimate physical admixture.

21. The composition of claim 1, further comprising neoeriocitrin, isonaringin, hesperidin, neodiosmin, naringenin, poncirin and rhiofolen.

22. An oral care composition having a pH of 3 to 8.5 comprising: (a) from 0.1 to 10% w/w based on the total weight of the composition, of a stock solution comprising (i) 0.45% to 9% of an extract or biomass comprising a mixture of water soluble bioflavonoids derived from the pith of the bitter orange wherein naringin and neohesperidin comprise in excess of 75% of the mixture of bioflavonoids and (ii) fruit acid; and (b) sodium hyaluronate of an average molecular weight between 800,000 and 4,000,000 daltons; (c) water; and (d) optionally a pharmaceutically acceptable carrier therefore.

23. The composition of claim 22, further comprising neoeriocitrin, isonaringin, hesperidin, neodiosmin, naringenin, poncirin and rhiofolen.

Description

EXAMPLE 1

Preparation of Stock Solution

(1) (a) Preparation of HPLC 45

(2) The starting material comprises the pith of immature, bitter (blood/red) oranges such as Seville oranges that are classed as inedible and from which the pips, flesh and oily skin have been substantially removed. The starting material is milled and then crushed in water or water/ethanol in a ratio of about 1:10-20 (solvent: starting material). The resulting mixture is filtered to leave a water-soluble biomass, which is retained, and an insoluble biomass, which is discarded. The water-soluble biomass is then subject to fine filtration, after which it is flash-distilled to leave a brown, hygroscopic powder (HPLC 45). Alternatively, the HPLC 45 is available from Exquim (Grupo Ferrer).

(3) (b) Bioflavonoid Composition of HPLC 45

(4) Analysis of the HPLC 45 obtained in step (a) shows that 45% of the total composition of HPLC 45 comprises bioflavonoids, with the balance (55%) comprising pectins, sugars and minor organic acids. The percentage (by weight of bioflavonoids in the HPLC 45) of the following bioflavonoids are present:

(5) TABLE-US-00004 % Bioflavonoids Bioflavonoid in HPLC 45 Isocriocirm 2.4 Isonaringin 2.7 Narangin 52.0 Hesperidin 3.1 Neohesperidin 27.8 Neodiomin 3.1 Naringenin 3.4 Poncirin 4.4 Rhiofolen 1.1 Total 100%

(6) Accordingly, by weight of the total composition of HPLC 45, the following bioflavonoids are present:

(7) TABLE-US-00005 Bioflavonoid % HPLC 45 Isocriocirm 1.1 Isonaringin 1.2 Narangin 23.4 Hesperidin 1.4 Neohesperidin 12.5 Neodiomin 1.4 Naringenin 1.5 Poncirin 2.0 Rhiofolen 2.8
(c) Preparation of Stock Solution

(8) TABLE-US-00006 Ingredient % Stock Solution HPLC 45 15 Citric acid 15 Malic acid 15 Ascorbic acid (vitamin C)* 5* Choline hydroxide solution (45% in water)* 15* Glycerine 15* Water 20* Total 100% [*Ascorbic acid and choline hydroxide can be replaced by choline ascorbate 5%, with amounts of glycerine and water increased to 25% each]

(9) The water, glycerine and ascorbic acid are blended together at ambient temperature and the temperature then increased to 50 deg C. The choline hydroxide is added to neutralize the ascorbic acid (starting pH=1.2; finishing pH=5.5-6.0).

(10) Then, the remaining acids (citric and malic) are added, followed by the HPLC 45, resulting in a stock solution having a pH of 6.2 to 7.2, and comprising 6.75% bioflavonoids (w/w of the stock solution).

EXAMPLE 2A

Mouthspray

(11) TABLE-US-00007 INGREDIENT PERCENTAGE Glycerine 10.000 Ethanol 15.000 Xylitol 2.000 Polysorbate 20 1.500 Stock solution, pH adjusted 1.000 to 6.44 Flavour 0.200 Sodium Saccharin 0.080 Cetyl Pyridinium Chloride 0.050 Disodium Phosphate 0.075 .12H.sub.2O Sodium Hyaluronate 0.005 Water q.v. to 100%
A. Laboratory Sample:

(12) A mouthspray according to the invention was prepared as follows, using the above-noted ingredients: In vessel (A) disperse the sodium hyaluronate in the water with stirring to give a lump free solution. Add the sodium saccharin, cetyl pyridinium chloride, disodium phosphate, xylitol and stock solution and stir until all the ingredients are fully dissolved. Add the glycerine and mix until homogeneous. In a separate vessel (B) combine the polysorbate 20, flavour and ethanol. Mix until the flavour is fully dispersed. Add the contents of vessel (B) to vessel (A) with stirring to give a homogeneous liquid.

(13) B. Alternative (Production) Method:

(14) A mouthspray according to the invention may be prepared as follows, using the above-noted ingredients: In vessel (A) disperse the sodium hyaluronate in the glycerine. With stirring, add the water to give a lump free solution. Add the xylitol, stock solution, sodium saccharin, cetyl pyridinium chloride and disodium phosphate, and stir until all the ingredients are fully dissolved. In a separate vessel (B) combine the polysorbate 20, flavour and ethanol. Mix until the flavour is fully dispersed. Add the contents of vessel (B) to vessel (A) with stirring to give a homogeneous liquid.

EXAMPLE 2B

Mouthspray

(15) TABLE-US-00008 INGREDIENT PERCENTAGE Glycerine 10.000 Ethanol 15.000 Xylitol 2.000 Polysorbate 20 1.500 Stock solution, pH adjusted to 6.44 1.000 Flavour 0.200 Sodium Saccharin 0.080 Cetyl Pyridinium Chloride 0.050 Disodium Phosphate .12H.sub.2O 0.075 Sodium Hyaluronate 0.01 (average molecular weight 1,500,000) Water q.v. to 100%

(16) A mouthspray according to the invention may be prepared as above using the above-noted ingredients.

EXAMPLE 3

Mouthrinse

(17) TABLE-US-00009 INGREDIENT PERCENTAGE Glycerine 25.000 Ethanol 0.000 Xylitol 2.000 Polysorbate 20 1.500 Stock solution (pH = 6.23 after 6 0.500 months at RTP) Flavour 0.200 Sodium Saccharin 0.050 Sodium Fluoride 0.050 Disodium Phosphate .12H.sub.2O 0.050 CI 18965 (Yellow 2G) 0.0009 CI 42051 (Patent Blue V) 0.0003 Sodium Hyaluronate 0.01 (average molecular weight 1,500,000) Water q.v. to 100% 70.53
A. Laboratory Sample:

(18) An oral rinse according to the invention may be prepared as follows, using the above-noted ingredients: In mixing vessel (A) disperse the sodium hyaluronate in the water with stirring to give a lump free solution. Add the sodium saccharin, sodium fluoride, disodium phosphate, colour(s), xylitol and stock solution and mix until all ingredients are fully dissolved. Add the glycerine and mix until homogeneous. In a separate vessel (B) combine the polysorbate 20 and flavour. Mix until the flavour is fully dispersed. Add the contents of vessel (B) to vessel (A) with stirring to give a homogeneous liquid.

(19) B. Alternative (Production) Method:

(20) An oral rinse according to the invention may be prepared as follows, using the above-noted ingredients: In mixing vessel (A) disperse the sodium hyaluronate in the water with stirring to give a lump free solution. Add the humectant(s), stock solution, sodium saccharin, sodium fluoride, disodium phosphate and colour. Mix until all ingredients are fully dissolved. In a separate vessel (B) combine the polysorbate 20, flavour and ethanol. Mix until the flavour is fully dispersed. Add the contents of vessel (B) to vessel (A) with stirring to give a homogeneous liquid.

EXAMPLE 4

Toothpaste

(21) TABLE-US-00010 INGREDIENT PERCENTAGE Glycerine 30.000 Hydrated Silica - abrasive 12.000 Hydrated Silica - thickening 11.000 Xylitol 10.000 Cocamidopropyl Betaine (30%) 3.000 Xanthan Gum 1.000 Stock solution (pH = 6.68 after 6 0.500 months at RTP) Sodium Hyaluronate 0.1 (average molecular weight 1,500,000) Flavour 1.000 Sodium Saccharin 0.260 Sodium Fluoride 0.240 Titanium Dioxide 0.500 Water to 100%
A. Laboratory Sample:

(22) A toothpaste according to the invention may be prepared as follows, using the above-noted ingredients: In mixing vessel (A) disperse the sodium hyaluronate in the water with stirring to give a lump free solution a and add the glycerine. To this add the sodium saccharin, sodium fluoride, stock solution and xylitol and stir to dissolve. Transfer the contents of vessel A to a vacuum mixer (vessel B). Preblend the powders (hydrated silicas, xanthan gum and titanium dioxide) in vessel C and add to the liquid phase in the vacuum mixer (B). Mix under vacuum until homogeneous. Add the surfactant and flavour to the vacuum mixer (B) and mix under vacuum to form a smooth paste.

EXAMPLE 5

Preparation of Stock Solution

(23) The following stock solution was prepared as above:

(24) TABLE-US-00011 Bioflavonoid mix 3.3% Malic acid 4.5% Citric acid 4.5% Glycerin 7.5% Ascorbic acid 1.5% Water 78.6% Ph of solution 1.5 to 1.75

EXAMPLE 6

Preparation of Stock Solution

(25) The following stock solution was prepared as above:

(26) TABLE-US-00012 Bioflavonoid mix 3.3% Malic acid 4.5% Citric acid 4.5% Choline ascorbate 6.0% LFG61 alkyl glycoside 13.3% Propylene glycol 7.5% Water 60.9% Ph of solution 1.5 to 1.75
Anti-Bacterial Activity

(27) Tests to determine whether compositions of the invention are active against a range of anaerobes and facultative bacteria that are implicated in periodontal disease may be conducted as follows.

(28) Method & Materials

(29) Bacteria that may be used include Actinomyces odontolyticus, Actinomyces viscosus, Porphyromonas gingivalis, Prevotella intermedia, Prevotella buccae, Prevotella dentalis, Streptococcus gordonii and Streptococcus sanguinis, in particular ATCC-type strains. All the anaerobes may be grown in Fastidious Anaerobic Broth (FAB) at 37 deg C. for 24 h in a Don Whitely Anaerobic Chamber (available from Don Whitely, Yorkshire, UK). The facultative bacteria may be grown in nutrient broth in 10% (v/v) carbon dioxide at 37 deg C. for 24 h. The starter culture is 1 ml of an overnight growth containing approximately 10.sup.6 cfu/ml. The cultures are supplemented with compositions of the invention in concentrations down to 1/10,000 (0.001%); the diluent being the appropriate broth. Growth may be estimated by a spectrophotometric increase in absorption at 650 nm.

(30) Growth on 5% (v/v) blood agar plates may be used to assess the toothpaste and mouthwash compositions. Wells approximately 0.5 cm diameter may be cut in the agar and filled with dilutions of the respective composition; the diluent being the appropriate broth. The plates are pre-inoculated with approximately 0.2 ml of broth containing 10.sup.6 cfu/ml. The minimum inhibitory concentration may be taken as the well concentration were no inhibition of growth is seen.

(31) Additional Tests

(32) Two stock solutions were tested: Example 5 and Example 6. Ranges of two-fold dilutions were prepared for each formulation, having a pH of 1.75 and 2.0, respectively, using either Blood-Heart Infusion (BHI) or Sabouraud's broth as the diluent, giving solutions with a range of concentrations (8%-0.015625% Stock Solution, v/v).

(33) Strains of several bacterial and candidal species (see Table 1) were obtained and cultured for 48 h under the appropriate conditions. Suspensions of each microorganism were prepared in broth (BHI broth for bacteria, Sabouraud's broth for Candida spp.) to a turbidity level approximately equal to MacFarland standard 3.0.

(34) TABLE-US-00013 TABLE 1 Species of microorganism used in this study Aerobic bacteria Anaerobic bacteria Yeast Streptococcus Actinomyces odontolyticus Candida albicans gordonii Streptococcus Actinomyces viscosus Candida dubliniensis sanguinis Clostridium difficile Candida glabrata Porphyromonas gingivalis Candida krusei Prevotella buccae Candida parapsilosis Prevotella intermedia Candida tropicalis

(35) A 100 l-volume of each microbial suspension was added to the wells of a microtitre plate. An equal volume of the solution of either Example 5 or Example 6 (containing the matching broth) was added to each well, giving final concentrations of Stock Solution of 4%-0.0078125% (v/v). Wells were also prepared containing no Stock Solution and/or no microorganism, to act as controls. Each microtitre plate was incubated for 24 h at 37 C. under the appropriate atmospheric conditions. After incubation the relative amounts of each microbial species were estimated by measuring the turbidity in each well using a spectrophotometer reading absorbance at a wavelength of 544 nm. Absorbance readings were blanked using the controls with an absence of microorganisms.

(36) The Minimal Inhibitory Concentration (SIC) was defined as the lowest concentration of Stock Solution that resulted in a significantly reduced amount of the microorganism (i.e. >50% reduction) in comparison to the controls where the microorganism was grown in the absence of Stock Solution. Experiments were performed in triplicate and MICs were determined for the formulations of Example 5 and Example 6 in the presence of each microbial species.

(37) Results:

(38) The MIC values observed for the 13 microorganisms in response to both formulations of Example 5 and Example 6 are summarised below in Table 2.

(39) The growth of each of the microorganisms studied was inhibited by both formulations of Example 5 and Example 6, with the exception of Candida glabrata. This yeast species' growth did not appear to be inhibited at all by Example 6 even when it was present at a concentration of 8% (v/v), the highest concentration used in this study.

(40) Comparison of the MICs from the two formulations of Example 5 and Example 6 suggested that Example 5 was more effective than Example 6 at inhibiting microbial growth. The MIC for each microorganism was lower with Example 5 than Example 6, with the exception of Porphyromonas gingivalis for which both formulations had an equal value of 1%. Furthermore, Example 5 inhibited the growth of each microorganism when at a concentration of 1% (v/v), even if the reduction of growth was not quite >50% (the criterion set in this study to define the MIC). This supports the use of 1% (v/v) of the stock solution of Example 5 as the preferred working concentration in future products and research.

(41) TABLE-US-00014 TABLE 2 MIC values observed in this study for each microorganism in response to the formulations of Example 5 and Example 6. MIC (% Stock Solution, v/v) Microorganism Example 5 Example 6 Actinomyces odontolyticus 0.015625 2 Actinomyces viscosus 2 4 Clostridium difficile 1 2 Porphyromonas gingivalis 1 1 Prevotella buccae 2 4 Prevotella intermedia 2 4 Streptococcus gordonii 0.03125 2 Streptococcus sanguinis 0.03125 2 Candida albicans 0.125 8 Candida glabrata 0.0625 >8* Candida krusei 0.015625 2 Candida parapsilosis 0.03125 8 Candida tropicalis 0.0625 8 *-No inhibition of growth apparent, even with highest concentration used in this study