This invention relates to products for the treatment of mammals suffering from gut disorders which may arise from nutritional, bacterial, viral or protozoal causes and lead to fluid depletion, acidosis, and imbalances or loss of essential electrolytes. These problems typically arise in immature animals such as calves and lambs.

The intestinal losses of water and electrolytes, including bicarbonate, are major factors in the pathogenesis of diarrhoea (scour) in calves. Calf scour continues to represent a major financial loss to the farmer and also a great source of concern to the veterinary profession. This invention is primarily intended to be applied in the treatment of calves with diarrhoea and description of the invention herein will be related thereto. It will, however, be appreciated that other animals, including humans, may also suffer from abnormal conditions leading to similar dehydration, acidosis and electrolyte loss for which these products may be used.

The causes of dehydration have been well researched and although many interacting factors must be considered it is observed that severe dehydration includes reduced fluid absorption as well as increased secretion and is predisposed by loss of sodium and accompanied by imbalance of other electrolytes in the extracellular fluids. Acidosis, where blood pH drops from a normal range (7.34-7.40) to a lower range (perhaps 6.85-7.15) is mainly attributable to increases in loss of bicarbonate ions, increases in lactic acid (due to poor tissue perfusion and anaerobic metabolism) and additional factors such as increases in organic acid production by intestinal bacteria.

Losses of sodium and bicarbonate ions are considered to be the most serious aspects of diarrhoea but losses of potassium and calcium are also observed. It is therefore necessary to provide a satisfactorily formulated oral rehydration solution (ORS) for the treatment of diarrhoea, both in humans and animals.

There are many products currently available for use in non-human animals which have varying degrees of efficacy with respect to replacement of electrolytes, correction of acidosis and provision of an energy source. The Applicant's own Life Aid P (trade mark) is one such rehydration product, whilst another is Lectade Plus (trade mark of SmithKline Beecham Animal Health) . Applicant's earlier invention in the same field is described in EP-A-0 644 767.

Whereas rehydration products may be administered intravenously, orally, or subcutaneously, the oral route is much preferred unless dehydration has already progressed to a serious level (8-10% or greater loss of bodyweight) when a combination of parenteral and oral administration should be employed. Conventional oral rehydration solutions are, however, formulated to address the dehydration, acidosis and electrolyte problems of diarrhoea but their use imposes virtual starvation because, in comparison to milk, the solutions are calorie deficient. Nutritional oral rehydration compositions aim to achieve energy levels more comparable with that of the natural milk denied to the animal during the use of an ORS.

An object of this invention is to provide an improved veterinary product providing effective rehydration whilst countering metabolic acidosis, calorific deficit and bodyweight loss more effectively than was previously possible, whilst also accelerating the recovery of enteric form and function.

A further object of this invention is to provide a product capable of supporting the structure and function of the intestine itself.

Accordingly this invention provides an oral rehydration composition, especially a nutritional oral rehydration composition, characterised by the presence of glutamine in an amount effective to enhance nutritional uptake. The composition of the invention is preferably a composition to be made up in water at point of use as an oral rehydration formulation, which composition comprises glutamine, an intimate mixture of metabolizable energy

source, electrolytes, and bicarbonate precursors, the concentration of sodium being about 120 millimoles per litre (mmol/1) of final formulation. The composition preferably also contains potassium chloride to obtain an ionically balanced product.

Preferably also the composition contains calcium chloride and magnesium acetate in order to provide free calcium and magnesium ions for ionic replacement. Preferably the composition contains differing bicarbonate precursors to increase the probability of utilisation. The precursors are provided as a plurality of physiologically acceptable carboxylic acid anions with corresponding physiologically acceptable cations including sodium and other alkali metals, the yield of bicarbonate obtainable in use being about 80 mmol/1 of final formulation. The most preferred group of precursors includes alkali metal, especially sodium, salts of a tricarboxylic acid and two other carboxylic acids. Propionate (propanoate) , acetate and citrate are the preferred bicarbonate precursors being converted to this anion in vivo to yield the desired concentration. Metabolism of theseprecursors also contributes to the energy yield of the present product.

The composition of this invention also contains a significantly greater level of energy source than the current market leading prior art compositions, preferably about 380 mmol/1, and this is preferably as glucose which also enhances the uptake of sodium. Glutamine is included at a concentration of about 30 mmol/1 to enhance the uptake of sodium ions, the absorption of glucose and as an additional energy source. The glutamine facilitates the metabolic utilisation of the absorbed glucose and also has beneficial effects on intestinal cells. The more effective use of glucose thus achieved avoids the need to consider even higher glucose content which could, at excessive concentration, draw water from the tissues into the gut and also provide a substrate for bacterial fermentation, both with detrimental effects.

The composition is preferably provided in particulate form, e.g. as a substantially dry powder or granules but may alternatively be provided as a liquid concentrate for dilution when required for administration. In the form of an intimate mixture of dry (typically not more than about 1.8 % w/w water content) components of this invention (which may include a desiccant) , the formulation is storage stable for extended periods.

The product may be produced for distribution as a substantially dry powder composition, or as a liquid concentrate for make up to a dosage formulation at point of use. Furthermore, this invention is suitable for packaging and supply as a single mix in a dosage quantity in a container such as a sachet or capsule, for example. Preferably the unitary dosage package contains components to be made up in water at point of use as an oral rehydration formulation comprising an intimate mixture of Dextrose anhydrous 75.7%w/w, Sodium chloride 3.4%w/w, Potassium chloride 2.5%w/w, Sodium citrate dihydrate 5.4%w/w, Sodium acetate 0.7%w/w, Sodium propionate l.l%w/w, Glutamine 4.9%w/w with the balance consisting of a dessicant and a colouring additive.

Advantageously the liquid concentrate when made up with water according to directions provided therewith yields a formulation in liquid form for oral administration comprising about 378.0 mmol/1 Dextrose, about 30.0 mmol/1 Glutamine, about 120.0 mmol/1 Sodium, about 16.7 mmol/1 Citrate, about 20 mmol/1 Acetate, about 10.0 mmol/1 Propionate, about 6.0 mmol/1 Magnesium, about 9.0 mmol/1 Calcium, about 100 mmol/1 Chloride, and about 30.0 mmol/1 Potassium.

The following examples illustrate the ionic strength of the invention when prepared for use.

Table 1 :

Solution compositions I II III IV (mmol/1) (mmol/1) (mmol/1) (mmol/1)

Sodium chloride 52.00 52.00 52.00 52.00

Potassium chloride 30.00 30.00 30.00 30.00

Sodium citrate 16.70 16.70 16.70 16.70

Sodium acetate 8.00 8.00 8.00 8.00

Sodium propionate 10.00 10.00 10.00 10.00

Calcium chloride 9.00 9.00 9.00 9.00

Magnesium acetate 6.00 6.00 6.00 6.00

Glucose 100.00 378.00 378.00 756.00

Glutamine 5.00 30.00 45.00 60.00

The invention will now be further described by way of the following example. (Example 1)

A trial was conducted in diarrhoeic calves to ascertain the benefit from the inclusion of glutamine in veterinary

ORS solutions. The trial was based on the methods published by Michell et al . (1992, Br. vet. J. , 148, 505-522, Appendix 1) .

Three test solutions were used in the trial, B, G and Y. The formulations of the solutions used are shown in Table 2.

Solution B represents a control solution. Solution G tested the effect of adding calcium and magnesium ions as the concentrations of these ions often fall in diarrhoeic calves, especially during the period of administration of oral rehydration therapy. The inclusion of these divalent cations would not be expected to have any other function than ionic replacement. Solution Y was to test for beneficial effects of glutamine addition.

Table 2 :

Solution compositions B G y (mmol/1) (mmol/1) (mmol/1)

Sodium chloride 40.00 52.00 52.00

Potassium chloride 20.00 30.00 30.00

Sodium citrate 16.70 16.70 16.70

Sodium acetate 20.00 8.00 8.00

Sodium propionate 10.00 10.00 10.00

Calcium chloride 0.00 9.00 9.00

Magnesium acetate 0.00 6.00 6.00

Glucose 378.00 378.00 378.00

Glutamine 0.00 0.00 30.00

The central tenet of oral rehydration therapy is that it minimises further fluid loss and helps replace fluid lost during the diarrhoeic process. Using the study design outlined below, two direct measurements of body fluid are feasible in a controlled study with full laboratory back up. The first of these is Plasma Volume, representative of the fluid in circulation in the bloodstream. The second is Extracellular Fluid which, in addition to plasma fluid, includes the fluid bathing cells in the animal's tissues. Blood volume can also be calculated from plasma volume and packed cell volume.

In the trial, after induction of diarrhoea, the calves received two litres of the relevant ORS twice daily for two days, i.e. four consecutive feeds. Thus, the data to the 48 hour timepoint represents the effect of administration of the pure ORS only. For the next four feeds, calves received two litres of 50% ORS/milk substitute solution (consistent with the recommended usage specified by most manufacturers) , with the exception of some calves from Groups B and Y which received the ORS alone for four days, i.e. eight consecutive feeds. Given that there was no significant difference in the body fluid data between these two subgroups for both solution B and Y, the data are combined for analysis. In the trial, the calves which showed no decline in both ECF and Plasma Volume following the induction of diarrhoea, prior to

the initiation of treatment, were excluded from the analysis of changes in ECF, Plasma Volume. The results of the trial are shown in Tables 2, 3, 4, 5 and 6. Table 3: Change in ECF Volume (litres) (± SE)

GROUP Pre- Pre¬ Post- Post- diarrhoea treatment treatment treatment

(0 hrs) (0-48 hrs) (0-96 hrs)

Group B 15.5 -0.5 -2.1 -1.5

(n=12) 0.7 0.3 0.5 0.3

Group Y 15.3 -1.4 -0.4 -0.6

(n=ll) 0.9 0.4 0.7 0.7

Group G 14.6 -1.0 -1.4 -1.1

(n=7) 0.6 0.9 1.1 1.3

Table 4: Change in Plasma Volume (litres) (± SE)

GROUP Pre- Pre¬ Post- Post- diarrhoea treatment treatment treatment

(0 hrs) (0-48 hrs) (0-96 hrs)

Group B 3.1 -0.4 -0.1 0.1

(n=12) 0.1 0.1 0.1 0.1

Group Y 3.0 -0.5 0.3 0.3

(n=ll) 0.2 0.1 0.1 0.1

Group G 2.8 -0.4 0.1 0.1

(n=7) 0.2 0.1 0.1 0.1

Table 5: Bodyweight (kg) (± SE)

GROUP Pre-treatment Post-treatment Post-treatment (0 hrs) (96 hrs) (0-96 hrs change)

Group B 44.25 41.92 2.3

(n=12) 2.2 2.3 0.4

Group Y 42.0 41.5 0.5

(n=10) 1.0 1.0 0.4

Group G 41.79 39.21 ^• 2.6

(n=7) 2.4 1.9 0.9

Table 6: Change in Blood Volume (litres) (± SE)

GROUP Pre- Pre¬ Post- Post- diarrhoea treatment treatment treatment

(0 hrs) (0-48 hrs) (0-96 hrs)

Group B 5.2 -0.5 -0.1 0.0

(n=12) 0.3 0.2 0.2 0.1

Group Y 4.9 -0.8 0.4 0.3

(n=ll) 0.3 0.1 0.1 0.1

Group G 4.8 -0.5 0.1 0.1

(n=7) 0.3 0.2 0.1 0.1

Table 7: Change in PCV (ml/dl) (± SE)

GROUP Pre- Pre¬ Post- Post- diarrhoea treatment treatment treatment

(0 hrs) (0-48 hrs) (0-96 hrs)

Group B 39.5 1.4 -0.3 -0.9

(n=12) 2.0 0.8 0.9 0.9

Group Y 38.0 1.5 -1.5 -1.6

(n=ll) 1.9 1.5 1.3 1.0

Group G 40.7 2.1 -0.6 -2.0

(n=7) 2.9 1.6 1.5 1.8

Solution Y was the only one to significantly increase blood volume within 48 hours (p < 0.01) and sustain the increase subsequently. It was also the only one to correct the packed cell volume (PCV) within 48 hours and sustain the recovery subsequently (Table 6) . These results are important because PCV, the proportion of blood occupied by red cells, is the main determinant of its viscosity; high viscosity seriously impedes capillary blood flow. Blood volume is a less direct measurement than plasma volume (from which it is derived, using PCV) but it is the determinant of circulatory 'fill' . The nutritional benefits of glutamine addition can clearly be seen from the results, especially the body weight data for these calves over the study period. Whilst all

three solutions contained the same amount of dextrose, as an energy source and to enhance sodium absorption, the group treated with solution Y demonstrated no statistically significant decrease in bodyweight over the period of treatment, whereas B and G did (p < 0.001, p < 0.05) . The above formulations constitute a single therapy for the treatment of a calf suffering from scour and address the areas of electrolyte replacement, fluid replacement, correction of acidosis and provision of an energy source, provided the appropriate dose is given and repeated as specified.

Example 2

A commercial product provided as a unitary dosage package to be made up at point of use as an oral rehydration formulation consists of an intimate mixture of Dextrose anhydrous 75.7%w/w, Sodium chloride 3.4%w/w, Potassium chloride 2.5%w/w, Sodium citrate dihydrate 5.4%w/w, Sodium acetate 0.7%w/w, Sodium propionate l.l%w/w, Glutamine 4.9%w/w with the balance consisting of a dessicant and a colouring additive.

Example 3

A similar product to be sold in liquid form to be prepared as a concentrate at a convenient volume which when maade up with water according to directions to be provided upon the package yields a formulation in liquid form for oral administration consisting of about 378.0 mmol/1 Dextrose, about 30.0 mmol/1 Glutamine, about 120.0 mmol/1 sodium, about 16.7 mmol/1 Citrate, about 20 mmol/1 Acetate, about 10.0 mmol/1 Propionate, about 6.0 mmol/1 Magnesium, about 9.0 mmol/1 Calcium, about 100 mmol/1 Chloride, and about 30.0 mmol/1 Potassium.

An advantage of the above-mentioned invention is that it can be used over a longer period at full strength compared with other ORS products. This is because the high calorie content in the product of the invention avoids

virtual starvation, again unlike those of the traditional prior art. Furthermore, because most of the calories in the product are derived from glucose, additional beneficial effects of glutamine on glucose utilisation can be seen. Because the formulation can be used for a longer period at full strength compared with other oral rehydration products, the glutamine can act for long enough to allow beneficial effects on gut cells (which are renewed in about three days) . In conjunction with there being no detrimental effects on body fluid levels, as previously described, there was, in fact, a positive effect on plasma glucose levels and body weight by the prolonged administration of the pure ORS. Calves treated with solutions B and Y for four days showed a rise of 3.1 + 1.0 mmol/1 in plasma glucose whereas those given 50% ORS/milk for the last four feeds had a rise of only 0.7 + 0.3 mmol/1. Similarly, those calves treated with solutions B and, Y for four days lost 0.5 + 0.5 kg, whereas those treated with 50% ORS/milk for the last four feeds lost 1.5 ± 0.5 kg bodyweight.