KR0177306B1 - Composition of mixture of somatotropin and vitamins - Google Patents

Abstract

The present invention relates to a mixture composition of somatotropin and vitamin having in vivo activity, wherein the mixture composition of somatotropin and vitamin, characterized in that the mixture of somatotropin and at least two fat-soluble vitamins, As a means of administration up to now, not only can it be injected into the body by the most common injection method, but also a synergistic effect can be obtained by simultaneously administering a separately administered vitamin preparation together with somatotropin, This can alleviate the pain and reduce the labor required for injection.

Description

Combination composition of somatotropin and vitamin

1 is a and b is a graph showing the weight gain by the composition of the present invention in dwarf rats,

2 is a graph showing the increase in acid flow rate by the composition of the present invention in cows.

The present invention relates to a combination composition of somatotropin and vitamins having in vivo activity, and in particular, by making an appropriate mixed composition of somatotropin and fat-soluble vitamins and administering them parenterally to an animal, it is necessary to separately administer them. The present invention relates to a composition designed to solve the problem and to obtain a sustained effect and a synergistic effect of two drugs.

The development of genetic engineering has made cattle somatotropins available for mass production by genetic recombination technology now commercially available for the purpose of increasing production, and pig somatotropins continue to improve feed efficiency and meat quality. Research is ongoing.

What has been developed so far to administer somatotropin with in vivo activity is most of the formulations that only prolong persistence by administering excess somatotropin to avoid the hassle of daily administration. For example, Korean Patent Publication No. 89-2631 discloses that oils may be added to vegetable oils that have been commonly used to make long-lasting formulations of drugs, such as antibiotics, by adding an antihydrating agent such as aluminum monostearate. A formulation wherein the persistence is only extended by homogenizing the gelation and somatotropin complexed to the transition metal is disclosed. As a similar example, European Patent No. 314,421 and Korean Patent Publication No. 87-1825 have disclosed formulations that extend only the sustainability of somatotropin with oil, which has also been applied to other drugs. These formulations are applied by replacing only somatotropin with the injection composition.

As a technique that does not use oil for sustainability, Korean Patent Application Nos. 90-1689 and 90-23104 disclose examples of using tocopherol acetate and excipients. However, at this time, tocopherol acetate is used as an excipient for prolonging only the persistence and is added in excess of the physiologically necessary dose, resulting in a problem of increasing the viscosity of tocopherol acetate in the winter when the temperature is low, which is inconvenient to administer.

In addition, most of the other techniques apply the solid compositions of the implant formulation to extend the persistence of somatotropin. As a representative technique, Korean Patent Publication Nos. 92-5660 and 90-6886 disclose that an expensive osmotic pump device is implanted through surgery, or a somatotropin produced by compression using a special implantation apparatus is implanted into an animal body. We tried to secure the sustainability. However, these transplant techniques are not only difficult to apply to animals, but also have a disadvantage of having a large sense of foreign object.

The present inventors have studied to solve this problem in consideration of the problems of somatotropin sustained-release formulation as mentioned above, and then formulated after mixing the fat-soluble vitamin in somatotropin in an appropriate ratio, as a means of administration so far In addition to being injected into the body through the most common injection methods, a single injection can be used to achieve a synergistic effect by administering these somatotropin and vitamin preparations at once.

In other words, the object of the present invention is not only can be injected into the body by the most common injection method as a means of administration so far, but also synergistic effects can be obtained by simultaneously administering the vitamin preparation previously administered together with somatotropin. The present invention also provides a somatotropin preparation that can reduce the number of injections of animals, thereby reducing pain, and at the same time reducing the labor required for injection administration.

In order to achieve the above object, the present invention provides a mixture composition of somatotropin and vitamins, characterized in that somatotropin and at least two fat-soluble vitamins are mixed.

Hereinafter, the present invention will be described in detail.

The vitamins mixed with somatotropin in the compositions of the present invention are, for example, fat-soluble vitamins such as vitamins A, D and E, which overcome the instability of protein drugs such as somatotropin when combined with water. It can be an advantageous drug for the preparation of mixed formulations.

In addition to the benefits of this formulation, vitamin A acts to enhance visual compliance, particularly dark adaptation, in relation to the photosensitizing rhodopsin and iodopsin present in the light-sensitive cells in the retina, the hepatic and cones. It also plays a role in improving abnormal dryness and degeneration of the mucous membranes, keratinization, damage, dry eye and corneal softening, and increased resistance to disease.In addition, it is essential for the maintenance of epithelial tissue, bone and tooth growth, and growth. It is also known to have an action.

Vitamin D is an anti-epileptic factor, which can lead to rickets, osteomalacia, osteopenia and tetany, especially when there is a need for vitamin D in pregnant or lactating animals and young animals, and insufficient intake from food. It needs to be administered by injection.

In addition, vitamin E helps normal reproductive processes in animals and prevents muscle developmental abnormalities, and can prevent encephalomalacia, muscle activity irregularities, muscle spasms, ataxia and tonic spasms. In addition, vitamin E is known to contribute to the prevention of mastitis, which is the most problematic in cows, mainly applied in the form of tocopherol acetate.

Of these fat-soluble vitamins, in particular vitamins A and D require caution because side effects can occur when applied in excess. Somatotropin is currently injected once every two weeks, so there is no side effect even with continuous administration, and it is necessary to prepare a dosage form to determine the dose to maximize the effects of these vitamins and the effect of somatotropin.

Somatotropin used in the composition of the present invention may be somatotropin of cattle or pigs, may be natural or produced by genetic recombination.

In the composition of the present invention, somatotropin is preferably contained in 20 to 35% by weight of the total composition, vitamin A is 500,000 to 2,000,000 units per gram of somatotropin, vitamin D per gram of somatotropin 75,000 to 300,000 units, and vitamin E is preferably mixed in a ratio of 1,200 to 3,500 units per gram of somatotropin.

In preparing the composition of the present invention, somatotropin may be homogenized with lecithin to make microparticles and then lyophilized, followed by homogeneous mixing with other vitamins, thereby preparing a combination composition of somatotropin and vitamin.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail based on an Example. However, the following examples are only examples of the present invention, and the present invention is not limited thereto.

Example 1

10 g of lecithin was added to 500 ml of bovine somatotropin (LG Chem. Biotech Research Institute) solution, mixed with a homogeneous mixer for 30 minutes, then placed in a fine grinder and ground until the particle size was 200 μm or less. I was. The suspension was sterilized through a 0.22 μm filter, placed in a freeze dryer and lyophilized. The freeze drying bottle was used for 3 days at a vacuum of 100 mtorr or lower and a temperature of −70 ° C. The freeze-dried powdered somatotropin showed 1.4% when the moisture content was measured by a heated moisture meter.

Separately, 267 g of vitamin A palmitate (1 g = 1,000,000 units, viscous solution, BASF) and 1 g of vitamin D ₃ (1 mg = 40,000 units, in powder form, Sigma) were weighed and dissolved using a magnetic bar.

To 6.7 g of freeze-dried bovine somatotropin, 9.5 g of vitamin E acetate (1 mg = 1.36 unit, viscous solution, ROCHE), and 5.03 g of the vitamin A and vitamin D ₃ solution prepared above were added and homogenized using a homogeneous mixer. After mixing, the mixture was placed in a vacuum chamber for 6 hours to remove bubbles from the composition.

In order to find out whether the composition prepared as described above is suitable for injection, the test was performed after storing at room temperature and 4 ° C. for one day. The injectability test measured the time that the composition exited the syringe when a 3 ml disposable syringe was filled with 2 ml of the present composition, fitted with a 15 gauge 3/4 inch needle and pressed with a 3 kg weight. Injectable results are shown in Table 1.

Example 2

The composition was prepared in the same manner as in Example 1, but 6.7 g of freeze-dried bovine somatotropin, 12 g of vitamin E acetate, and 2.5 g of a mixture of vitamin A and vitamin D ₃ were mixed. Injectable results are shown in Table 1.

Example 3

To prepare a composition in the same manner as in Example 1, 7g of 6.7g vitamin E acetate and 7.5g of a mixture of vitamin A and vitamin D ₃ was used as lyophilized bovine somatotropin. Injectable results are shown in Table 1.

Example 4

The composition was prepared in the same manner as in Example 1, but 6.7 g of freeze-dried bovine somatotropin, 4.5 g of vitamin E acetate, and 10 g of a mixture of vitamin A and vitamin D ₃ were used. Injectable results are shown in Table 1.

[Comparative Example]

To prepare a composition in the same manner as in Example 1, only 6.7g of lyophilized bovine somatotropin, and 14.5g of vitamin E acetate were used. Injectable results are shown in Table 1.

As shown in Table 1, in the case of Examples 1 to 4 in which somatotropin was mixed with at least two vitamins, the injection ability was good at low temperature as well as at room temperature, but the comparative example in which only tocopherol acetate was mixed was used at low temperature. The injection performance was so poor that it could interfere with the administration to the animals.

Example 5

Animal experiments were carried out using the composition prepared as in Examples 1 to 4 and Comparative Example using porcine somatotropin instead of bovine somatotropin, using a dwarf rat genetically induced dwarf rat It was.

Dwarf rats were females 8 weeks old, weighing about 100 g, and were fed freely with water and feed. Weighing for 3 days prior to administration of the composition to the rat was used as the reference weight of each individual, 0.04 ml (10 mg equivalent of pig somatotropin equivalent) of each composition was injected into the rat subcutaneous subcutaneous, 7 after administration Weighed at a constant time each day for days. As a control, 4 dwarf rats that received nothing were weighed during the test period. The cumulative weight gain accumulated after administration of the composition is shown in Table 2.

1a and 1b is a graph showing the increase in weight by the composition of the present invention in dwarf rats.

As shown in Table 2 and Figure 1, the composition of the present invention showed a weight gain of about 40% before and after 7 days compared to the control.

Example 6

Using the compositions prepared in Example 1 and Comparative Example, an increased milk flow test was performed on 24 milking Holstein cows.

24 cows were divided into 8 groups each in consideration of maternity and milking days, and 500 mg (250 mg / ml) of the bovine somatotropin formulation mixed with the vitamin ADE of Example 1, the control group which was not administered at all, was administered. One group and the other group received the same dose injection of the formulation of the comparative example. Dosing was injected subcutaneously in both tail caudal humerus, four times at two week intervals. The daily milk flow was measured for 2 weeks before administration, and the base oil flow rate of each individual was measured. The results are shown in Table 3.

2 is a graph showing the increase in acid flow rate by the composition of the present invention in cows.

As shown in Table 3 and Figure 2, the composition of the present invention showed an increase in milk flow of about 30% compared to the control at 7 to 8 weeks of administration.

On the other hand, the composition of somatotropin mixed with only tocopherol acetate, the weight gain effect in the dwarf rats and the effect of increasing the acid flow rate in the cow was similar to the composition of the present invention, but was not preferable in terms of poor injection ability.

As described above, the mixture composition of the present invention comprising somatotropin and at least two vitamins has good injection ability even at room temperature or at low temperature, and shows that it has an excellent effect on promoting growth and enhancing the yield of cow. there was.

In addition, considering that the somatotropin and vitamin complex formulations are separately injected when used in real animals, the combination composition of the somatotropin and the complex vitamin of the present invention can solve this inconvenience, and As is already known, by administering a multi-vitamin to the animal can also benefit the health of the target animal.

Claims (9)

A mixture composition of somatotropin and vitamins, characterized in that somatotropin and at least two fat-soluble vitamins are mixed. The composition of claim 1 wherein the somatotropin is bovine or porcine somatotropin. The composition of claim 1 wherein the somatotropin is somatotropin produced by genetic recombination. The composition of claim 1 wherein the somatotropin is somatotropin homogenized with lecithin. The composition of claim 1 wherein the somatotropin is 20-35% by weight of the composition. The composition of claim 1, wherein the fat soluble vitamin is vitamin A, vitamin D or vitamin E. 7. The composition of claim 6, wherein the vitamin A is mixed at 500,000 to 2,000,000 units per gram of somatotropin. 7. The composition of claim 6, wherein the vitamin D is mixed at 75,000 to 300,000 units per gram of somatotropin. 7. The composition of claim 6, wherein the vitamin E is mixed at 1,200 to 3,500 units per gram of somatotropin.