Composition comprising non-neutralised tocol phosphate and a vitamin A compound

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 formulation comprising a composition and a hydrophobic delivery vehicle, wherein: the composition comprises a non-neutralised tocol phosphate and a vitamin A compound, wherein the non-neutralised tocol phosphate is a combination of a non-neutralised mono-(tocopheryl) phosphate and a non-neutralised di-(tocopheryl) phosphate; and the hydrophobic delivery vehicle is an oil or a wax, and is present in an amount of at least about 80.0% w/w of the total concentration of the formulation, wherein the oil is selected from the group consisting of vegetable oils, fruit oils, seed oils, grain oils, and nut oils.

2. The formulation according to claim 1 wherein the pH of the non-neutralised tocol phosphate is less than about 4, about 2, about 3, about 2 to about 4, or about 2 to about 3.

3. The formulation according to claim 1 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.

4. The formulation according to claim 1 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.

5. The formulation according to claim 1 wherein the vitamin A compound is a carotenoid.

6. The formulation according to claim 5 wherein the carotenoid is selected from the group consisting of alpha-carotene, beta-carotene, gamma-carotene, beta-cryptoxanthin and lycopene.

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

8. The formulation according to claim 1, wherein the composition comprises a combination of a mono-(tocopheryl) phosphate and a di-(tocopheryl) phosphate, and beta-carotene.

9. The formulation according to claim 1, wherein the composition further comprises a nutrient compound.

10. The formulation according to claim 9 wherein the nutrient compound is selected from the group consisting of antioxidants, vitamins, minerals and trace elements.

11. The formulation according to claim 10 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.

12. The formulation according to claim 11 wherein the nutrient compound is selected from the group consisting of: vitamin D2; vitamin D3; , , , and -tocopherols; , , and -tocotrienols; vitamin K1; vitamin K2; vitamin K3; vitamin B1; vitamin B2; vitamin B3; vitamin B6; vitamin B12; catechins; reserveratrol; proanthocyanidins; genistein; daidzein; lycopene; lutein; seaxanthin; linoleic acid; conjugated linoleic acid; linolenic acid; docosahexaenoic acid; and eicosapentaeonic acid.

13. The formulation according to claim 1, wherein the hydrophobic delivery vehicle is an oil selected from the group consisting of canola oil, coconut oil, corn oil, cottonseed oil, olive oil, Olea europaea leaf extract, palm oil, peanut oil, rapeseed oil, safflower oil, sesame oil, sunflower oil, almond oil, cashew oil, hazelnut oil, macadamia oil, Macadamia integrifolia (macadamia) seed oil, mongongo nut oil, pecan oil, pine nut oil, pistachio oil, walnut oil, bottle gourd oil, buffalo gourd oil, pumpkin seed oil, watermelon seed oil, acai oil, blackcurrant seed oil, borage seed oil, evening primrose oil (Oenothera biennis oil), amaranth oil, apricot oil, apple seed oil, argan oil, artichoke oil, avocado oil (Persea gratissima oil), babassu oil, ben oil, borneo tallow nut oil, cape chestnut oil, cocoa butter, carob pod oil, cocklebur oil, cohune oil, coriander seed oil, dika oil, false flax oil, flax seed oil, grape seed oil (Vitis vinifera), hemp oil, kapok seed oil, lallemantia oil, marula oil, meadowfoam seed oil, mustard oil, nutmeg butter, okra seed oil, orange oil, papaya seed oil, perilla seed oil, pequi oil, pine nut oil, poppyseed oil, prune kernel oil, quinoa oil, ramtil oil, rice bran oil, rose hip oil (Rosa eglanteria oil), royle oil, sacha inchi oil, sandalwood oil (Santalum spicatum oil), tea seed oil, thistle oil, tomato seed oil and wheat germ oil.

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 Treatment Week 0) 311,000 Previous 5 (reading taken 28 weeks before Treatment Week 0) 128,400 Previous 4 (reading taken 23 weeks before Treatment Week 0) 222,000 Previous 3 (reading taken 17 weeks before Treatment Week 0) 161,000 Previous 2 (reading taken 12 weeks before Treatment Week 0) 322,444 Previous 1 (reading taken 7 weeks before Treatment Week 0) 271,000 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 Treatment Week 0) 1,491,000 Previous 5 (reading taken 28 weeks before Treatment Week 0) 604,857 Previous 4 (reading taken 23 weeks before Treatment Week 0) 221,375 Previous 3 (reading taken 17 weeks before Treatment Week 0) 234,000 Previous 2 (reading taken 12 weeks before Treatment Week 0) 560,111 Previous 1 (reading taken 7 weeks before Treatment Week 0) 1,122,667 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/rd, 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.