Composition

Abstract

Compositions and formulations comprising a non-neutralized tocol phosphate and a vitamin A compound, which are suitable for the treatment of inflammation and/or infection in breast or udder tissue, more particularly in a mammary gland, reducing the somatic cell count in a lactating subject and supplementing vitamin E levels in a subject.

Claims

1. A method for reducing somatic cell count in a lactating subject comprising administering a formulation to the lactating subject, wherein the formulation comprises a non-neutralised tocol phosphate, a vitamin A compound, and a hydrophobic delivery vehicle, wherein the non-neutralised tocol phosphate is a combination of a non-neutralised mono-(tocopheryl) phosphate and a non-neutralised di-(tocopheryl) phosphate.

2. A method for treating inflammation and/or infection in breast or udder tissue comprising administering a formulation to a subject in need thereof, wherein the formulation comprises a non-neutralised tocol phosphate, a vitamin A compound, and a hydrophobic delivery vehicle, wherein the non-neutralised tocol phosphate is a combination of a non-neutralised mono-(tocopheryl) phosphate and a non-neutralised di-(tocopheryl) phosphate.

3. The method of claim 2 wherein the breast or udder tissue is a mammary gland.

4. The method of claim 2 wherein the inflammation and/or infection is mastitis.

5. The method of claim 2 wherein the subject is selected from humans, domestic mammals including companion animals, working animals, livestock, and zoological/park mammals.

6. The method of claim 2 wherein the combination of the non-neutralised mono-(tocopheryl) phosphate and the non-neutralised di-(tocopheryl) phosphate comprises: (a) a compound selected from the group consisting of non-neutralised mono-(tocopheryl) phosphate, non-neutralised mono-(tocopheryl) phosphate monosodium salt, non-neutralised mono-(tocopheryl) phosphate disodium salt, non-neutralised mono-(tocopheryl) phosphate monopotassium salt, non-neutralised mono-(tocopheryl) phosphate dipotassium salt, or a combination of any thereof; and (b) a compound selected from the group consisting of non-neutralised di-(tocopheryl) phosphate, non-neutralised di-(tocopheryl) phosphate monosodium salt, and non-neutralised di-(tocopheryl) phosphate monopotassium salt, or a combination of any thereof.

7. The method of claim 2 wherein the ratio (w/w %) of non-neutralised mono-(tocopheryl) phosphate to non-neutralised di-(tocopheryl) phosphate is about 4:1 to about 1:4.

8. The method of claim 2 wherein the non-neutralised tocol phosphate has a pH of less than about 4.

9. The method of claim 2 wherein the non-neutralised tocol phosphate comprises from about 0.01% w/w up to about 40% w/w of the total concentration of the composition.

10. The method of claim 2 wherein the vitamin A compound is selected from the group consisting of vitamin A, a derivative of vitamin A, a metabolite of vitamin A, a precursor of vitamin A, or pro-vitamin A, or a combination thereof.

11. The method of claim 2 wherein the vitamin A compound is a carotenoid.

12. The method of claim 11 wherein the carotenoid is selected from the carotene class including alpha-carotene, beta-carotene, gamma-carotene, beta-cryptoxanthin and lycopene.

13. The method of claim 2 wherein the ratio (w/w %) of non-neutralised tocol phosphate to vitamin A compound is from about 0.01:1 to about 100:1.

14. The method of claim 2 wherein the vitamin A compound is beta-carotene.

15. The method of claim 2 wherein the formulation further comprises a nutrient compound selected from the group consisting of antioxidants, vitamins, minerals and trace elements.

16. The method of claim 15 wherein the nutrient compound is selected from the group consisting of coenzyme Q10, ubiquinol, vitamin D compounds, vitamin E compounds, vitamin K compounds, folic acid, vitamin B compounds, vitamin C, vitamin P, vitamin F, lutein, zeaxanthin, cysteine, flavonoids, isoflavones, bilberry, ginkgo biloba, grape seed extract, phytonutrients, alpha lipoic acid, bilberry, bioflavinoids, unsaturated fatty acids, calcium, phosphorus, magnesium, fluorine, phosphorus, sulfur, sodium, potassium, chloride, calcium, iodine, cobalt, copper, iron, manganese, molybdenum, selenium, zinc, chromium, cadmium, fluorine, nickel, silicon, tin, vanadium, niacin, and combinations thereof.

17. The method of claim 2 wherein the hydrophobic delivery vehicle is an oil or a wax.

18. The method of claim 2 wherein the hydrophobic delivery vehicle is present in an amount of at least about 60.0% w/w of the total concentration of the formulation.

19. The method of claim 2 wherein the non-neutralised tocol phosphate has a pH of about 2 to about 4.

Description

FIGURES

(1) The invention is described further by way of example with reference to the accompanying drawings in which:

(2) FIG. 1A is a bar chart showing the average somatic cell count of the dairy cows of Group A over a period of time prior to treatment with Formulation A (represented in grey) and over a 2 week treatment period (represented in black).

(3) FIG. 1B is a bar chart showing the percentage change in the average somatic cell count of the dairy cows of Group A over a period prior to treatment with Formulation A and over a 2 week treatment period.

(4) FIG. 2A is a bar chart showing the average somatic cell count of the dairy cows of Group B over a period of time prior to treatment with Formulation B (represented in grey) and over a 5 week treatment period (represented in black).

(5) FIG. 2B is a bar chart showing the percentage change in the average somatic cell count of the dairy cows of Group B over a period of time prior to treatment with Formulation B and over a 5 week treatment period.

EXAMPLES

(6) The invention is described further by reference to the following non-limiting examples of the invention.

Example 1: Preparation of Formulations

(7) The following formulations of the present invention were prepared for oral delivery.

(8) TABLE-US-00002 Component Formulation A Formulation B Non-neutralised TPM 1% w/w (~0.5 g) 2% w/w (~1 g) Beta-carotene 0.1% w/w (~50 mg) 0.2% w/w (~100 mg) Olive oil balance to 100% balance to 100% (total ~60 ml) (total ~60 ml)

(9) Each of Formulations A and B were prepared by combining non-neutralised TPM (pH about 2 to about 4) with olive oil, and stirring the mixture at about 70 C. The mixture was then cooled to below 40 C. before the beta-carotene was added, with stirring.

Example 2: Treatment of Dairy Cows Suffering Mastitis

(10) 35 dairy cows showing signs of either sub-clinical or clinical mastitis with persistent elevated somatic cell counts, were kept separated from the main herd (consisting of approximately 300 head). Various attempts to treat the mastitis in the cattle had been made, and the mastitis was considered problematic.

(11) 18 of the 35 dairy cows were selected based on their somatic cell counts; their somatic cell counts and previous antibiotic treatment was documented. The 18 dairy cows were divided into 2 groups consisting of 9 dairy cows each and designated Group A and Group B.

(12) Final antibiotic treatment ceased at least 4 weeks prior to commencing treatment with formulations of the present invention. A baseline or treatment week 0 somatic cell count was measured.

(13) At treatment week 0, the Group A dairy cows had elevated somatic cell counts but not severe, with an average somatic cell count of 429,000 cells/ml, whereas the Group B dairy cows showed some clinical signs of mastitis and generally had much higher somatic cell counts with an average somatic cell count of 1,243,222 cells/ml.

(14) The Group A dairy cows were treated with Formulation A of Example 1 and the Group B dairy cows were treated with Formulation B of Example 1.

(15) Both groups were treated daily using an oral drench, administered each morning after milking. The oral drench was delivered via a drenching gun that delivered approximately 60 ml of the relevant formulation.

(16) Each week, a milk sample was collected from each of the dairy cows in the respective groups and somatic cell counts were measured for each individual dairy cow.

(17) Group A Results

(18) The Group A dairy cows were treated for 2 weeks. The results of this study are shown in the table below.

(19) TABLE-US-00003 TABLE 1 Somatic cell counts for the Group A dairy cows Average Average percentage somatic cell change in count somatic cell Time point (cells/ml) count Previous 6 (reading taken 33 weeks before 311,000 Treatment Week 0) Previous 5 (reading taken 28 weeks before 128,400 Treatment Week 0) Previous 4 (reading taken 23 weeks before 222,000 Treatment Week 0) Previous 3 (reading taken 17 weeks before 161,000 Treatment Week 0) Previous 2 (reading taken 12 weeks before 322,444 Treatment Week 0) Previous 1 (reading taken 7 weeks before 271,000 Treatment Week 0) Treatment Week 0 429,000 Treatment Week 1 341556 23% Treatment Week 2 222,111 45%

(20) The results shown for the somatic cell counts are an average of the somatic cell counts of the 9 dairy cows for this group. The percentage change values were calculated as the difference in the somatic cell count compared to the somatic cell count at treatment week 0, and expressed as a percentage of the somatic cell count for that treatment week.

(21) The graph shown in FIG. 1A shows the decrease in somatic cell count over the treatment period. After 2 weeks of daily treatment, the average somatic cell count decreased from 429,000 cells/ml to 222,111 cells/ml, which correlates to a 45% reduction in the somatic cell count after 2 weeks of treatment.

(22) Group B Results

(23) The Group B dairy cows were treated for 5 weeks. The results of this study are shown in the table below.

(24) TABLE-US-00004 TABLE 2 Somatic cell counts for the Group B dairy cows Average Average percentage somatic cell change in count somatic cell Time point (cells/ml) count Previous 6 (reading taken 33 weeks before 1,491,000 Treatment Week 0) Previous 5 (reading taken 28 weeks before 604,857 Treatment Week 0) Previous 4 (reading taken 23 weeks before 221,375 Treatment Week 0) Previous 3 (reading taken 17 weeks before 234,000 Treatment Week 0) Previous 2 (reading taken 12 weeks before 560,111 Treatment Week 0) Previous 1 (reading taken 7 weeks before 1,122,667 Treatment Week 0) Treatment Week 0 1,243,222 Treatment Week 1 2,247,333 109% Treatment Week 2 1,145,444 12% Treatment Week 3 1,265,556 7% Treatment Week 4 852,625 33% Treatment Week 5 516,556 54%

(25) The results shown for the somatic cell counts are an average of the somatic cell counts of the 9 dairy cows for this group. The percentage change values were calculated as the difference in the somatic cell count compared to the somatic cell count at treatment week 0, and expressed as a percentage of the somatic cell count for that treatment week.

(26) The graph shown in FIG. 2A shows the decrease in somatic cell count over the treatment period. After 5 weeks of daily treatment, the average somatic cell count decreased from 1,243,222 cells/ml to 516,556 cells/ml, which correlates to a 54% reduction in the somatic cell count after 5 weeks of treatment.

(27) This example clearly demonstrates the effectiveness of formulations of the present invention in the reduction of somatic cell count and therefore treatment of mastitis in dairy cows.

Example 3: Vitamin E Supplementation in Dairy Cows Suffering Mastitis

(28) The following are results of vitamin E levels in 4 dairy cows showing signs of mastitis treated with a formulation of the present invention after 1 week. The blood of the 4 dairy cows was tested at a commercial pathology laboratory.

(29) TABLE-US-00005 Baseline Vitamin E Supplementation (mol/L) Vitamin E (mol/L) Change % Change DC1 13.6 13.9 +0.3 +2.2 DC2 9.6 10.8 +1.2 +12.5 DC3 14.6 15.2 +0.6 +4.1 DC4 16.1 16.7 +0.6 +3.7

Example 4: Formulations Comprising Nutrients

(30) Further embodiments of an oral drench can be prepared according to Example 1, but comprising additional nutrients, such as for example, selenium, copper, zinc, a vitamin E compound. The nutrients are added at the same time as the beta-carotene.

(31) Specifically designed oral drenches would be administered to dairy cattle as described in Example 2, and compared with dairy cattle administered with an oral drench not comprising any nutrients. The blood levels of vitamin E could also be measured as in Example 3.

Example 5: Optimisation of Administration Route

(32) Formulations as described in Example 4 could also be prepared using suitable excipients as intramammary infusions or topical creams. These could be compared against the effect of the drench formulation as outlined in Examples 2, 3 and/or 4.

(33) Many modifications may be made to the embodiments and examples described above without departing from the spirit and scope of the invention.

(34) It is to be understood that, if any prior art publication is referred to herein, such reference does not constitute an admission that the publication forms a part of the common general knowledge in the art, in Australia or any other country.

(35) In the claims which follow and in the preceding description of the invention, except where the context requires otherwise due to express language or necessary implication, the word comprise or variations such as comprises or comprising is used in an inclusive sense, i.e. to specify the presence of the stated features but not to preclude the presence or addition of further features in various embodiments of the invention.