NZ272317A

NZ272317A - Veterinary formulation; comprises calcium borogluconate, vitamin b12 or a derivative thereof and a carrier; treatment of milk fever in animals

Info

Publication number: NZ272317A
Application number: NZ27231795A
Authority: NZ
Inventors: Donald George Mclaren; Wayne Frederick Leech
Original assignee: Bomac Lab Ltd
Priority date: 1995-06-09
Filing date: 1995-06-09
Publication date: 1997-07-27
Also published as: AU718893B2; GB9612133D0; AU5588296A; GB2302019A; GB2302019B

Description

New Zealand Paient Spedficaiion for Paient Number £72317 272317 Patents Form No. 5 Our Ref: PZ500824 NEW ZEALAND PATENTS ACT 1953 Complete After Provisional No. 272317 F:'led: 9 June 1995 COMPLETE SPECIFICATION FORMULATIONS FOR TREATMENT OF MILK FEVER We, BOMAC LABORATORIES LIMITED, a New Zealand company of Cnr Wiri Station Road & Hobill Avenue, Manukau City, Auckland, New Zealand hereby declare the invention, for which We pray that a patent may be granted to us and the method by which it is to be performed, to be particularly described in r .a by the following statement: 272317 FORMULATIONS FOR TREATMENT OF MILK FEVER This invention relates to veterinary formulations comprising vitamin B12 and calciurr, borogluconate for the treatment of milk fevar and/cr vitamin Bi2 or cobalt deficiency in cattle and sheep. It also relates to a method of manufacture of such formulations, and use of such formulations in veterinary medicine.

Milk fever (also known as hypocalcaemia, parturient paresis, periparturient paresis or parturient apoplexy) is an afebrile disease of mature dairy cows and ewes. It occurs most commonly at or soon after parturition. It is usually associated with the sudden onset of profuse lactation and is accompanied by a decrease in serum calcium levels. Serum magnesium may also be depressed. The disease is manifested by circulatory collapse, generalised paresis and depression.

Current methods of treatment are directed towards the restoration of serum calcium levels to normal. This must be done as soon as possible to avoid muscular and nervous damage and recumbency. Calcium is given to the animal usually in the form of calcium salts either as calcium borogluconate, calcium gluconate or calcium chloride. The preferred route is by intravenous injection but calcium borogluconate may be given subcutaneously, and calcium chloride orally. 272317 In order that an early return to normal metabolism is enhanced, any supplementation to treatment with calcium which stimulates the supply of energy is desirable.

In ruminants, the volatile free fatty acid, propionic acid, is a major source of energy. The metabolism of propionic acid is interfered with by a deficiency of vitamin B]_2• Vitamin B^2 (cyanocobalamin or hydroxocobalamin) is a cobalt-containing vitamin required by cells throughout the body for conversion of ribose nucleotides into deoxyribose nucleotides, a major step in the formation of deoxyribonucleic acid (DNA). Thus it is an essential nutrient for nuclear maturation and cell division. A deficiency in vitamin B12 results in a form of anemia. However, anemias from deficiencies of vitamin B12 are less apparent clinically in domestic animals compared with humans.

Adult ruminants are not dependent on a dietary source of vitamin B^2 because bacteria within the rumen synthesise all the supplies of vitamin B12 needed. However, cobalt is required by the ruminal micro-organisms to synthesise this vitamin Bi2» an<* a deficiency in cobalt can, therefore, cause a deficiency of the vitamin. 2723^ Formulations comprising calcium borogluconate are well known. However, formulations comprising both calcium borogluconate and vitamin B^2 for the treatment of milk fever are novel. Moreover, formulations containing vitamin B12 which are stable at relatively low pH (e.g. pH 3-4) are also novel.

An object of the invention is to provide a treatment for animals suffering from milk fever and a formulation for such treatment or to at least provide the public with a useful choice.

Other objects of the invention may become apparent from the following description, given by way of example only.

According to one aspect of the present invention, there is provided a veterinary formulation comprising calcium borogluconate and vitamin B^2 or a derivative thereof with a pharmaceutically acceptable carrier.

Preferably, cyanocobalamin may be employed in the formulation, and the formulation may be prepared as an aqueous solution for administration by subcutaneous injection.

In a preferred form, the veterinary formulation may have a pH in the range 3.0 to 4.0, and more preferably 3.3 to 3.8. - ni^1 According to a further aspect of the present invention, there is provided a method of treatment of animals suffering from milk fever and/cr vitamin or cobalt deficiency with a formulation of calcium borogluconate and cyanocobalamin in a pharmaceutically acceptable diluent.

This method of treatment is particularly directed at ruminant animals, especially cattle and sheep.

According to a further aspect of the invention, there is provided a method of producing a formulation comprising calcium borogluconate and cyanocobalamin for veterinary use, said method comprising dissolving said cyanocobalamin in an aqueous solution of calcium borogluconate at a pH of substantially 3.0 to 4.0.

Other aspects of the invention will become apparent from the following description which is given by way of example only, and with reference to the accompanying examples.

The calcium borogluconate may be prepared by the combination of boric acid and calcium gluconate (providing calcium in the form of: the calcium borogluconate complex) . The formulation may also comprise a preservative such as sodium benzoate. Other ingredients may also be added. 27 2^ A surprising feature of the formulations of this invention is their stability, and in particular, the stability of certain types of vitamin B^2 at lower pH values (3 to 4).

Due to different potency of active ingredients and to ameliorate the decline in potency in a complex chemical matrix, an excess or overage is recommended in pharmaceutical formulations. In view of the unstable nature of vitamins, an overage is essential. Vitamins are hygroscopic and relatively more stable in dry (pure crystal) form. Optimum stability of vitamin B12 in aqueous solution is in the pH range 4.5-5 (Merck Index, pl7l0, 1993). Such solutions are also sensitive to temperature and reducing agents. The present inventors have tested their formulations employing vitamin overages of 30%, 50% and 100% and found 50% the most appropriate concentration.

Embodiments of the invention will now be described, by way of example only. 2723^7 7 FORMULATION 1 INGREDIENT QUANTITY Boric Acid Calcium gluconate Sodium benzoate Cyanocobalamin* Water for injection 40.OOOg 210.OOOg 1.OOOg 0 .03g 1.000 litre [*An overage (50%) of Cyanocobalamin is included to allow for the moisture content of raw material and to ensure the label claim is met.] To make a 1200L batch, 800L of water for injection is heated to boiling. The boric acid, sodium benzoate and calcium gluconate are added and dissolved by mixing. The volume is made up with water for injection, and the solution is heated for 30 minutes at 95°C. It is left to stand overnight before the addition of the cyanocobalamin, with mixing to dissolve. The volume is then made up again with water for injection.

A feature of the formulation is its stability during storage. Tests were conducted at a variety of temperatures over 21-month period during which time samples were analysed at regular intervals. The physical and chemical characteristics of the formulation were monitored. No significant changes in physical characteristics were 272317 displayed. Except for cyanocobalamin, the chemical characteristics showed only minor fluctuations, as shown in the following Table I. Cyanocobalamin levels showed a decrease in the first 3 months, but were stable thereafter, although somewhat lower in the samples stored at 30°C.

TABLE I STABILITY DATA POR FORMULATION 1 Time After Manufacture Initial 3m 6m 12m 18m 21m Stored At 20"C pH* 3.8 3.8 3.8 3.8 3.8 3.8 Boric Acid (% w/v) 3.65 3.80 3.73 3.71 3.74 3.73 Calcium {% w/v) 1.77 1.82 1.75 1.81 1.86 1.78 Cyanocobalamin (mcg/ml) 28.0 20.3 23.0 24.3 20.7 23.5 Stored At 25°C pH* 3.8 3.8 3.8 3.8 3.8 3.8 Boric Acid (% w/v) 3.65 3.81 3.75 3.79 3.70 3.73 Calcium (% w/v) 1.77 1.80 1.75 1.82 1.82 1.77 Cyanocobalamin (mcg/ml) 28.0 21.9 23.4 25.3 20.6 24.2 Stored At 30°C pH* 3.8 3.8 3.8 3.8 3.8 3.8 Boric Acid (% w/v) 3.65 3.82 3.77 3.78 3.67 3.75 Calcium (% w/v) 1.77 1.80 1.75 1.82 1.8 0 1.77 Cyanocobalamin (mcg/ml) 28.0 21.5 21.9 22.9 17.7 20.6 [♦measured at 19 or 20°C] 272317 9 FORMULATION 2 MANUFACTURING LABEL FORMULATION CLAIM Boric Acid 60.OOOg 3 75mg/mL (37.5%) Calcium Borogluconate Calcium Gluconate Sodium Benzoate Cyanocobalamin 315.OOOg 1.OOOg 0.030g 20mcg/mL Water for iniection qs to 1.000 Litre Calcium Glucori=»t-e and Boric Acid forms the Calcium Borogluconate complex.

A 50% overage of Cyanocobalamin is added to compensate for the moisture and to ensure label claim is met.

Stability data carried out on Formulation 2 are shown in Table II. Again, there was no significant change in the physical characteristics of the product stored at 20 to 3 0°C for 21 months. There were only minor fluctuations in chemical characteristics, except for cyanocobalamin the levels of which declined in the first 3 months, but were stable thereafter when stored at 20°C or 25°C (within the product specification of 18 to 33 mcg/ml after 21 months). 2723^ At 30°C, there was about a 53% loss of cyanocobalamin after 21 months. Therefore, storage at below 25°C is desirable, as is shielding from sunlight.

TABLE II STABILITY DATA FOR FORMULATION 2 Time After Manufacture Initial 3m 6m 12m 18m 21m Stored At 20°C pH* 3 .6 3 .6 3 . 6 3 . 6 3 .6 3 . 7 Boric Acid (% w/v) .71 .84 . 84 . 75 . 62 . 62 Calcium (% w/v) 2 .71 2 .69 2 . 75 2 . 72 ? .73 2 . 73 Cyanocobalamin (mcg/ml) 21 19 22 Stored At 25°C pH* 3 .6 3 .6 3 . 6 3 .6 3 .6 3 .7 Boric Acid (% w/v) .71 .80 . 94 .80 .73 . 70 Calcium (% w/v) 2 .71 2 .69 2. 77 2 . 73 2 .79 2 . 75 Cyanocobalamin (mcg/ml) 24 22 Stored At 30°C pH* 3 .6 3 .6 3. 6 3 .6 3 .6 3 . 7 Boric Acid (% w/v) .71 .80 . 93 .86 .78 .78 Calcium (% w/v) 2 .71 2 .71 2. 68 2 .72 2 .75 2 . 74 Cyanocobalamin (mcg/ml) 22 22 12 14 [♦measured at 19 or 20°C] 272317 FORMULATION 3 Boric I-id Calcium Gluconate Magnesium Chloride Cyanocobalamin Sodium Benzoate MANUFACTURING LABEL FORMULATION CLAIM 40 .OOOg 210.OOOg 80.OOOg 0.030g 1.OOOg 250mg/mL (25%) Calcium Borogluconate 80mg/mL (8%] 20mcg/ml Water for inj ection qs to 1.000 Litre 1. Calcium Gluconate and Boric Acid forms the Calcium Borogluconate complex. 2. An overage (50%) of Cyanocobalamin is added to allow for the moisture content (e.g., 11% potency (e.g., 85-90%)) of raw material and also for the possible degradation in the formulated product.

Stability data carried out on Fcrmulatior 3 are shown in Table III. The physical characteristics of the formulation remained unchanged after 12 months stored at 20°C or 25°C. Boric acid, calcium/magnesium and chloride 272317 levels showed only minor fluctuations, whilst cyanocobalamin levels were also well maintained over the 12-month period of the tests.

TABLE III STABILITY DATA FOR FORMULATION 3 Time After Manufacture Initial 3m 6m 12m Stored At 20°C pH* 3.4 3.3 3.5 Boric Acid (% w/v) 4.05 4.07 3 . 97 Calcium/Magnesium (mmoL/L) 907 908 915 Chloride (% w/v) 2.96 2.95 2.97 Cyanocobalamin a 37b (mcg/ml) 31 28 . 5 Stored At 25°C pH* 3.4 3.4 3.4 3.5 Boric Acid (% w/v) 4.05 4.04 4.09 3.98 Calcium/Magnesium (mmoL/L) 907 917 901 927 Chloride (% w/v) 2.96 2.94 2 . 86 2.96 Cyanocobalamin a 37b (mcg/ml) 31 27.8 a Overage (100%) included when formulated, k Validated method of analysis.

* Measured at 19 or 20°C. 8723^7 FORMULATION 4 MANUFACTURING LABEL FORMULATION CLAIM Boric Acid 40.OOOg 250mg/mL (25%) Calcium Borogluconate Calcium Gluconate 210.OOOg Dextrose Monohydrate 275.OOOg 250mg/mL (25%) Dextrose Magnesium Chloride 40.OOOg 40mg/mL (4%) Magnesium Chloride Sodium Benzoate l.OOOg Cyanocobalamin Acetate 0.030g 20mcg/mL Water for injection qs to 1.000 Litre 1. Calcium Gluconate and Boric Acid forms the Calcium Borogluconate complex. 2. Dextrose Monohydrate contains 90.9% Dextrose 275 x 0.909 = 250g/L Dextrose. 272317 3. A 50% overage of cyanocobalamin Acetate is added to compensate for the moisture and to ensure label claim is met.

It has been surprisingly found that cyanocobalamin is stable at the acidic pH of the formulations: about pH 3.7 to 3.8 for Formulation 1 and slightly lower for Formulations 2 and 3.

A number of different types of vitamin B12 have been tested in metabolic formulations by the inventors and cyanocobalamin was found to be the most stable.

The target concentration of cyanocobalamin of the above formulations is about .002%, but concentrations down to .001% have been employed. Moreover, it is anticipated that concentrated formulations, containing up to 5% cyanocobalamin, may be employed.

These formulations may be administered by subcutaneous injection, preferably into the anterior half of the neck or over the ribs, in cattle or sheep (for example) suffering from milk fever, and/or as an energy stimulant in animals deficient in vitamin B^-

Claims

272317 - 15 - The invention therefore provides a unique way of delivering two important medications to an animal. Anecdotal evidence suggests that the metabolic formulations of the present invention enhance appetite in afebrile milk fever suffering cows. Although this invention has been described by way of example, it will be appreciated that improvements and/or modifications maybe made thereto without departing from the sprit and scope of the present invention as defined in the following claims. For example, although treatment of milk fever and vitamin B12 deficiency are described, the formulation may be applicable to many other diseases in animals. Also, although the invention has been described with particular reference to sheep and cattle, its use for the treatment of other animals is also envisaged. gl/specs/500824 272317 - 16 - CLAIMS: WHAT 1/W'E CLAIM is

1. A veterinary formulation comprising calcium -borogluconate, vitamin B-^2 or a derivative thereof and a pharmaceutically acceptable carrier.

2. A veterinary formulation comprising calcium borogluconate, cyanocobalamin and a pharmaceutically acceptable diluent.

3. A veterinary formulation according to either claim 1 or claim 2 wherein the pH is from substantially 3.0 to 4.0.

4. A veterinary formulation according to claim 3 wherein the pH is from substantially 3.3 to 3.8.

5. A veterinary formulation according to any one of the preceding claims further comprising a preservative.

6. A veterinary formulation according to any one of the preceding claims wherein the calcium borogluconate is in aqueous solution at a concentration of at least 25% w/v.

7. A veterinary formulation according to any one of the preceding claims further comprising magnesium chloride. 272317 - 17 -

8. A veterinary formulation according to claim 7 wherein the concentration of magnesium chloride is in the range 4 to 10% w/v.

9. A veterinary formulation according to any one of the preceding claims further comprising dextrose monohydrate.

10. A veterinary formulation according to any one of the preceding claims adapted for use in the treatment of milk fever and/or vitamin B-±2 deficiency in animals.

11. A method of treating animals suffering from milk fever comprising administering a veterinary formulation of any one of claims 1 to 9.

12. A method of treatment of animals suffering from milk fever and/or vitamin B]_2 or cobalt deficiency, said method comprising administering a veterinary formulation of any one of claims 1 to 9.

13. A method of producing a formulation of calcium borogluconate and cyanocobalamin for veterinary use, said method comprising dissolving said cyanocobalamin in an ajueous solution of calcium borogluconate at a pH of substantially 3.0 to 4.0. - 18 - ?7 2 3 1 7

14. A method according to claim 13 wherein the aqueous solution contains at least 25% calcium borogluconate w/v.

15. A method according to the claim 13 wherein the aqueous solution is substantially 25% w/v calcium borogluconate and further comprises magnesium chloride 4 to 10% w/v.

16. A veterinary formulation substantially as herein described and with reference to the examples given as formulations 1, 2, 3 and 4.

17 . A method of treatment of milk fever and/or vitamin B12 or cobalt deficiency in animals substantially as herein described with reference to examples given as formulations 1, 2, 3 and 4.

18. A method of producing a veterinary formulation substantially as herein described and with reference to the accompanying examples of formulations 1, 2, 3 and 4. END OF CLAIMS BOMAC LABORATORIES LIMITED By Their Attorneys JAMES & WELLS may 189?