Injectable composition of 25-hydroxy-vitamin d3

Abstract

The present invention relates to the use of 25-hydroxy-vitamin D3 in animal breeding. More particularly, the invention relates to an injectable composition of 25-hydroxy- vitamin D3 and its use of 25-hydroxy-vitamin D3 raising plasma25-hydroxy-vitamin D3 levels of piglets in pig breeding.

Description

[0001] NOVEL COMPOSITION

[0002] The present invention relates to the use of 25-hydroxy-vitamin D3 in animal breeding. More particularly, the invention relates to an injectable composition of 25-hydroxy-vitamin D3 and its use of 25-hydroxy-vitamin D3 raising the vitamin D status (measured by plasma25-hydroxy-vitamin D3 levels) of pigs, preferably newborn piglets, in pig breeding.

[0003] Vitamin D is an important nutrient for pigs, necessarily required to maintain bone formation and development in association with calcium and phosphorus. However, pigs in confinement housing are born with low serum vitamin D concentrations, which may result in vitamin D deficiency and related problems, such as even nursery death loss, leading to the postulate that modern production practices result in vitamin D-deficient pigs at weaning.

[0004] Piglets have the lowest level of serum vitamin D at birth (Horst and Littledike, 1982). Nursing pigs in outdoor settings can acquire vitamin D via two sources - one synthesized by sunlight and the other from sow’s milk, whereas pigs in confinement housing can acquire vitamin D through only sow’s milk. However, sow’s milk contains little vitamin D and placental transport of vitamin D is poor. Nonetheless, because fetal and nursing pigs are provided vitamin D via placental transfer and sow’s milk, the maternal vitamin D status is extremely important to supply adequate amount of vitamin D for piglets.

[0005] Any slight improvement in health, and therefore productivity, will have a major significant positive impact on the overall profitability of the swine enterprise. Therefore, it is important to enhance vitamin D status of pigs and piglets in breeding. Preventing vitamin D deficiency can prevent various health issues, including rickets, stiffness, weak and bent bones, and posterior paralysis and osteoporosis, poor bone development and improve overall health in young pigs. This is commonly achieved by intramuscular injection of vitamin D3, increasing serum 25-hydroxycholecalciferol (25-OH D3 or 25-hydroxy-vitamin D3) concentrations. Goff et al. (1984) therefore suggested that the parenteral vitamin D3 treatment before parturition by intramuscular injection of vitamin D3 to sows at 20 days pre-partum was an effective method for enhancing vitamin D status of piglets because the vitamin D status of sows is closely correlated to that of fetus and neonatal piglets.

[0006] Several recent studies have been conducted to investigate dietary vitamin D supplementation in form of Vit D3 or 25OHD3 to sows and piglets (Lauridsen et al., 2010; Witschi et al., 2011; Flohr et al., 2012; Rortvedt et al., 2012). The supplementation of vitamin D to gestation and lactation diets was demonstrated to improve serum vitamin D concentration of sows (Lauridsen et al., 2010).

[0007] However, the serum Vit D concentration of newborn piglets during the lactation period independently of the level of dietary Vit D supplementation on the sows are rather low (Thayer et al, 2019, summary below).

[0008] In accordance with the present invention it has surprisingly been found that administering 25-hydroxy-vitamin D3 at 5000 III bioequivalent D3 III to pigs, instead of vitamin D3 at 10 OOOIll which is fat soluble (slower absroption), by intramuscular injection, circulating levels of 25OHD3 were higher compared to vitamin D3 after 19 days of injection with a single injection.

[0009] Thus, in one aspect, the invention relates to the use of 25-hydroxy-vitamin D3 as an injectable for raising vitamin D status evaluated by plasma25-hydroxy-vitamin D3 levels of pigs in pig breeding.

[0010] In still another aspect the invention relates to a method of raising plasma25-hydroxy-vitamin D3 levels of pigs in pig breeding, wherein 25-hydroxy-vitamin D3 is administered to pigs as an injectable.

[0011] In another aspect the invention relates to an injectable composition of 25-hydroxy-vitamin D3 for raising plasma25-hydroxy-vitamin D3 levels of pigs in pig breeding. The term “suckling piglets” or “piglets” as used herein refers to a young pig still taking milk from its mother. This term is used for pigs from birth up to weaning (usually between 21 to 28 days old).

[0012] For the purposes of the invention, 25-hydroxy-vitamin D3 is suitably by intramuscular injection. The injectable composition may be formulated by dissolving 25-hydroxy-vitamin D3 , e.g., as a commercial formulation such as available under the Trademark ROVIMIX® Hy D® 1.25%, in water or preferably saline. 10 mg of 25-hydroxy-vitamin D3 equate to 1000IU. The injectable composition may further comprise other minerals, vitamins, amino acids and trace elements and nutrients.

[0013] For the purposes of the invention, 25-hydroxy-vitamin D3 is administered at least once. The administration of 25-hydroxy-vitamin D3 may be performed at any time, at any age, at any point in the life and in all stages of production of a pig, such as to reproductive animals, boars, pregnant, lactating and non-lactating sows, suckling and weaned piglets, growing and finishing pigs. Preferably, injection of 25-hydroxy-vitamin D3 is admistered after birth of the newborn piglets, preferably after 24 to 72 hours post birth.

[0014] Generally, 25-hydroxy-vitamin D3 is administered to pigs in an amount required to administer from 2.5 mg to 350 mg, preferably 5 mg to 300 mg of 25-hydroxy-vitamin D3 animal.

[0015] In one embodiment, the injectable may contain 100 to 12500 III 25-hydroxy-vitamin D3, preferably 200 to 12500 IU 25-hydroxy-vitamin D3, preferably 300 to 12500 IU 25-hydroxy-vitamin D3, preferably 400 to 12500 IU 25-hydroxy-vitamin D3, preferably 500 to 12500 IU 25-hydroxy-vitamin D3, preferably 600 to 12500 IU 25-hydroxy-vita-min D3, preferably 700 to 12500 IU 25-hydroxy-vitamin D3, preferably 800 to 12500 IU 25-hydroxy-vitamin D3, preferably 900 to 12500 IU 25-hydroxy-vitamin D3, preferably 1000 to 12500 IU 25-hydroxy-vitamin D3, preferably 1000 to 12000 IU 25-hy-droxy-vitamin D3, preferably 1000 to 11500 IU 25-hydroxy-vitamin D3, preferably 1000 to 11000 IU 25-hydroxy-vitamin D3, preferably 1000 to 10000 IU 25-hydroxy-vitamin D3 per mL injectable. More specifically, the injectable composition may comprise 25-hydroxy-vitamin D3 in a concentration of at least 0.00025 wt%, preferably at least 0.0003 wt%, preferably at least 0.00035 wt%, preferably at least 0.0004 wt%, preferably at least 0.00045 wt%, preferably at least 0.0005 wt%, preferably at least 0.00055 wt%, preferably at least 0.0006 wt%, preferably at least 0.00065 wt%, preferably at least 0.0007 wt%, preferably at least 0.00075 wt%, preferably at least 0.0008 wt%, preferably at least 0.00085 wt%, preferably at least 0.0009 wt%, preferably at least 0.00095 wt%, preferably at least 0.001 wt%, preferably at least 0.0015 wt%, preferably at least 0.002 wt%, preferably at least 0.0025 wt%, preferably at least 0.003 wt%, preferably at least 0.0035 wt%, preferably at least 0.004 wt%, preferably at least 0.0045 wt%, preferably at least 0.005 wt%, preferably at least 0.0055 wt%, preferably at least 0.006 wt%, preferably at least 0.0065 wt%, preferably at least 0.007 wt%, preferably at least 0.0075 wt%, preferably at least 0.008 wt%, preferably at least 0.0085 wt%, preferably at least 0.009 wt%, preferably at least 0.0095 wt%, preferably at least 0.01 wt%.

[0016] The injectable composition may comprise 25-hydroxy-vitamin D3 in a concentration of below 0.035 wt%, preferably below 0.03 wt%, preferably below 0.025 wt%, preferably below 0.02 wt%, preferably below 0.015 wt%.

[0017] The injectable composition may comprise 25-hydroxy-vitamin D3 in a concentration in the range from 0.00025 to 0.035 wt%, preferably in the range from 0.00025 to 0.03 wt%, preferably in the range from 0.00025 to 0.025 wt%, preferably in the range from 0.0003 to 0.025 wt%, preferably in the range from 0.0003 to 0.02 wt%, preferably in the range from 0.00035 to 0.02 wt%, preferably in the range from 0.0004 to 0.02 wt%, preferably in the range from 0.0004 to 0.015 wt%, preferably in the range from 0.00045 to 0.015 wt%, preferably in the range from 0.0005 to 0.015 wt%, preferably in the range from 0.0006 to 0.015 wt%, preferably in the range from 0.0007 to 0.015 wt%, preferably in the range from 0.0008 to 0.015 wt%, preferably in the range from 0.0009 to 0.015 wt%, preferably in the range from 0.001 to 0.015 wt%, preferably in the range from 0.001 to 0.01 wt%. Figures

[0018] Figure 1: Plasma levels of 25-hydroxy-vitamin D3 (ng / mL), just before the injection (D1 ), 6 days after the injection (D7) and 20 days post-injection (D21 ) with 25-hydroxy-vitamin D3 (HyD inj 5000 III) and vitamin D3 (Vit D3 inj 10000 IU) compared to the control (placebo).

[0019] The efficiency of the treatment in raising plasma levels of 25-hydroxy-vitamin D3 in pigs in accordance with the invention can be seen from the example.

[0020] Examples

[0021] Example 1

[0022] Experimental design and treatments

[0023] A total of 12 litters from pregnant sows receiving the same diet (with 2000 IU of Vit D3 inclusion in feed) were used for this experiment.

[0024] 1 newborn pig per sow were processed at birth (within 24h) and assigned to one of the three treatments indicated in the table below meaning 12 piglets / treatment. After 24h birth, all piglets including in the trial are subjected to a standard iron injection in one part of the neck and a second injection on the other side of the neck as indicated in the tables below. The products were injected at room temperature.

[0025]

[0026] Processing of the piglets involved weighing, ear-tagging, and gender identification. All the adoptions needed happened before the randomization.

[0027] Once animals were selected, BEFORE application of the treatments a blood sample was collected.

[0028]

[0029]

[0030] Treatments randomization

[0031] The treatment randomization per litter will be based on the body weight and gender in order to have a balance between treatments.

[0032] Creep feed to the piglets

[0033] No creep feed was administrated to the piglets during the lactation period from the litters in the study, what means that the only source of nutrients was from sows’ milk.

[0034] EXPERIMENTAL MEASURES AND ANALYSIS

[0035] Health status

[0036] Daily mortality and any reaction post injection were also recorded.

[0037] 24h post injection:

[0038] • Individual temperature of all the animals using traditional thermometer (as the current infrared thermometer is not accurate)

[0039] • Check and score if there is any local reaction in both side of the neck (left and right) where the injection was applied:

[0040] o Score 0: Normal

[0041] o Score 1 : Redness skin

[0042] o Score 2: Abscess (red, warm and swallow)

[0043] Vitamin D status

[0044] An assessment of vitamin D status of the piglets was done measuring the circulating level of 25(OH)D in blood using a UHPLC-MS / MS system.

[0045] Samples were collected:

[0046] • D1 old: just before the injection

[0047] • D7 old: 6 days after the injection

[0048] • D21 old: 20 days post-injection and just before the weaning moment.

[0049] RESULTS

[0050] Health status

[0051] No mortality during the whole trial

[0052] No local reaction in the neck 24h post-injection

[0053] No fever 24h post-injection was detected. Fever considered as rectal temperature >39.8°C according Manual Merk

[0054] (https: / / www.msdvetmanual.com / special-subjects / reference-guides / norrnal-rectal-temperature-ranges) Vitamin D status

[0055] Results on circulating 25OHD3 used commonly to determine the vitamin D status of the animals presented in the Table and Figure 1 , showed that 1 days after birth (day 1 old), newborn piglets had very low levels of 25OHD3.

[0056] 6 days later the treatments application, circulating 25OHD3 increased significantly when Vit D3 and 25OHD3 were injected compared to the control animals.

[0057] However, and as expected, the levels of 25OHD3 decreased 20 days after the injection, but interestingly when the injection was applied using the form of 25OHD3 the circulating levels of 25OHD3 were higher than vit D3.

[0058]

[0059] < < CONCLUSIONS

[0060] The novel application of vitamin D in the form of 25OHD3 in newborn piglets is a safe and efficient way to raise the vitamin D status (circulating 25OHD3) during the lactation period.

Claims

Claims1. The use of 25-hydroxy-vitamin D3 for raising plasma25-hydroxy-vitamin D3 levels of pigs in pig breeding, wherein 25-hydroxy-vitamin D3 is administered to pigs as an injectable.

2. The use as in claim 1 wherein 25-hydroxy-vitamin D3 is administered to pigs in an amount required to administer from 2.5 mg to 350 mg, preferably 5 mg to 300 mg of 25-hydroxy-vitamin D3 animal.

3. The use as in claim 1 wherein 25-hydroxy-vitamin D3 is administered to pigs in an amount of 1 mg to 125 mg (100 to 12500 III) 25-hydroxy-vitamin D3, preferably 2 mg to 125 mg (200 to 12500 IU) 25-hydroxy-vitamin D3, preferably 3 mg to 125 mg (300 to 12500 IU) 25-hydroxy-vitamin D3, preferably 4 mg to 125 mg (400 to 12500 IU) 25-hydroxy-vitamin D3, preferably 5 mg to 125 mg (500 to 12500 IU) 25-hydroxy-vitamin D3, preferably 6 mg to 125 mg (600 to 12500 IU) 25-hydroxy-vitamin D3, preferably 7 mg to 125 mg (700 to 12500 IU) 25-hydroxy-vitamin D3, preferably 8 mg to 125 mg (800 to 12500 IU) 25- hydroxy-vitamin D3, preferably 9 mg to 125 mg (900 to 12500 IU) 25-hydroxy- vitamin D3, preferably 10 mg to 125 mg (1000 to 12500 IU) 25-hydroxy-vitamin D3, preferably 10 mg to 120 mg (1000 to 12000 IU) 25-hydroxy-vitamin D3, preferably 10 mg to 115 mg (1000 to 11500 IU) 25-hydroxy-vitamin D3, preferably 10 mg to 110 mg (1000 to 11000 IU) 25-hydroxy-vitamin D3, preferably 10 mg to 100 mg (1000 to 10000 IU) 25-hydroxy-vitamin D3 per mL injectable.

4. The use as in any one of claims 1 - 3 wherein the pig is a reproductive animal, such as a sow or a boar, or a piglet.

5. A method for raising plasma25-hydroxy-vitamin D3 levels of pigs in pig breeding which comprises administering to the pig an effective amount of 25-hy- droxy-vitamin D3, wherein 25-hydroxy-vitamin D3 is administered as an injectable.

6. The method as in claim 5 wherein 25-hydroxy-vitamin D3 is administered in an amount from 2.5 mg to 350 mg, preferably 5 mg to 300 mg of 25-hydroxy- vitamin D3 animal.

7. The method as in claim 5 wherein the injectable contains 25-hydroxy-vitamin D3 in an amount of 1 mg to 125 mg (100 to 12500 III) 25-hydroxy-vitamin D3, preferably 2 mg to 125 mg (200 to 12500 IU) 25-hydroxy-vitamin D3, preferably 3 mg to 125 mg (300 to 12500 IU) 25-hydroxy-vitamin D3, preferably 4 mg to 125 mg (400 to 12500 IU) 25-hydroxy-vitamin D3, preferably 5 mg to 125 mg (500 to 12500 IU) 25-hydroxy-vitamin D3, preferably 6 mg to 125 mg (600 to 12500 IU) 25-hydroxy-vitamin D3, preferably 7 mg to 125 mg (700 to 12500 IU) 25-hydroxy-vitamin D3, preferably 8 mg to 125 mg (800 to 12500 IU) 25- hydroxy-vitamin D3, preferably 9 mg to 125 mg (900 to 12500 IU) 25-hydroxy- vitamin D3, preferably 10 mg to 125 mg (1000 to 12500 IU) 25-hydroxy-vitamin D3, preferably 10 mg to 120 mg (1000 to 12000 IU) 25-hydroxy-vitamin D3, preferably 10 mg to 115 mg (1000 to 11500 IU) 25-hydroxy-vitamin D3, preferably 10 mg to 110 mg (1000 to 11000 IU) 25-hydroxy-vitamin D3, preferably 10 mg to 100 mg (1000 to 10000 IU) 25-hydroxy-vitamin D3 per mL injectable.

8. The method as in any one of claims 5 - 7 wherein 25-hydroxy-vitamin D3 is administered to a reproductive animal, such as a sow or a boar, or a piglet.

9. An injectable composition of 25-hydroxy-vitamin D3 for raising plasma25-hy- droxy-vitamin D3 levels of pig in pig breeding.

10. The composition of claim 9, wherein the composition is a liquid composition of 25-hydroxy-vitamin D3.

11. The composition of any of claims 9 or 10, wherein the composition comprises 25- hydroxy-vitamin D3 in water.

12. The composition of any of claims 9 or 10, wherein the composition comprises 25- hydroxy-vitamin D3 in saline.

13. The composition of any of claims 9 to 12, wherein the concentration of 25-hy- droxy-vitamin D3 is at least 0.00025 wt%, preferably at least 0.0003 wt%, preferably at least 0.00035 wt%, preferably at least 0.0004 wt%, preferably at least 0.00045 wt%, preferably at least 0.0005 wt%, preferably at least 0.00055 wt%, preferably at least 0.0006 wt%, preferably at least 0.00065 wt%, preferably at least 0.0007 wt%, preferably at least 0.00075 wt%, preferably at least 0.0008 wt%, preferably at least 0.00085 wt%, pref-erably at least 0.0009 wt%, preferably at least 0.00095 wt%, preferably at least 0.001 wt%, preferably at least 0.0015 wt%, preferably at least 0.002 wt%, preferably at least 0.0025 wt%, preferably at least 0.003 wt%, preferably at least 0.0035 wt%, preferably at least 0.004 wt%, preferably at least 0.0045 wt%, preferably at least 0.005 wt%, preferably at least 0.0055 wt%, preferably at least 0.006 wt%, preferably at least 0.0065 wt%, preferably at least 0.007 wt%, preferably at least 0.0075 wt%, preferably at least 0.008 wt%, preferably at least 0.0085 wt%, preferably at least 0.009 wt%, preferably at least 0.0095 wt%, preferably at least 0.01 wt%.

14. The composition of any of claims 9 to 12, wherein the concentration of 25-hy- droxy-vitamin D3 is below 0.035 wt%, preferably below 0.03 wt%, preferably below 0.025 wt%, prefera-bly below 0.02 wt%, preferably below 0.015 wt%.

15. The composition of any of claims 9 to 12, wherein the concentration of 25-hy- droxy-vitamin D3 is in the range from 0.00025 to 0.035 wt%, preferably in the range from 0.00025 to 0.03 wt%, pref-erably in the range from 0.00025 to 0.025 wt%, preferably in the range from 0.0003 to 0.025 wt%, preferably in the range from 0.0003 to 0.02 wt%, preferably in the range from 0.00035 to 0.02 wt%, preferably in the range from 0.0004 to 0.02 wt%, preferably in the range from 0.0004 to 0.015 wt%, preferably in the range from 0.00045 to 0.015 wt%, preferably in the range from 0.0005 to 0.015 wt%, preferably in the range from 0.0006 to 0.015 wt%, preferably in the range from 0.0007 to 0.015 wt%, preferably in the range from 0.0008 to 0.015 wt%, preferably in the range from 0.0009 to 0.015 wt%,preferably in the range from 0.001 to 0.015 wt%, preferably in the range from 0.001 to 0.01 wt%.* * *

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