The short term regulation of blood glucose levels after meals or during the first few hours of starvation is mainly determined by the release of the hormones insulin or glucagon into the blood stream.

The initial high blood glucose levels after consuming carbohydrate-rich foods can often be followed by low blood glucose levels as the released insulin overcompensates, an effect known as a hypoglycaemic dip.

Impairments in the secretion or action of insulin gives rise to increases in blood glucose and can lead to the development of diabetes mellitus. There are three types of diabetes mellitus. Type I diabetes is caused by a lack of insulin as a result of damage to the B-cells of the pancreas and is treated with insulin administered by injection. Type 11 diabetes commonly occurs with age and includes defects in the ability of the B-cells to respond to increased glucose concentrations or decreased sensitivity of target cells to normal insulin levels. It has been hypothesised that controlling the incidence of insulin release might reduce the risk or postpone the onset of Type 11 diabetes. Type III diabetes occurs during pregnancy.

The standard dietetic approach to treating diabetes is to advise patients to reduce the consumption of high glycaemic index foods (i. e. those causing high blood glucose levels), to increase consumption of complex carbohydrates, and to spread their carbohydrate intake throughout the day. However, a balance between carbohydrate intake, other dietary components (mono-unsaturated fatty acids), exercise and insulin dose (Type I) or response must be maintained to avoid

hypoglycaemia, where blood glucose levels are low and which in severe cases can cause coma.

Trehalose (a-D-glucopyranosyl-a-D-glucopyranoside) is a naturally occurring non-reducing disaccharide found in fungi, certain yeasts, certain drought resistant plants and in the blood of insects. However, it has hitherto contributed an insignificant part of most human diets.

G. G. Birch in Process Biochemist, July 1970, page 9, briefly reviews the role of trehalose in nature. The author notes that trehalose is quite sweet-tasting, and if it were to be absorbed slowly or not at all by the body, without any ill-effect, it could provide a useful non-fattening dietetic or diabetic sugar.

It has subsequently been established that trehalose is, in fact, rapidly absorbed by the body. The mechanism of absorption is that trehalase enzyme in the microvilli of the small intestine breaks the trehalose down into its constituent glucose monomers, which are then absorbed through the intestinal wall.

W096/08979 describes isotonic or hypotonic sports beverages containing trehalose. The sports beverages containing trehalose are said to be useful for providing a quick energy boost by restoring blood glucose levels of athletes. The use of trehalose rather than glucose is said to be desirable because of the lower osmotic pressure of a given weight concentration of a disaccharide such as trehalose compared to a monosaccharide such as glucose. There is no suggestion that the sports beverages would be suitable for nutrition of diabetics.

EP-A-0619951 also describes energy supplements containing trehalose.

Data are presented showing that relatively small orally administered doses of trehalose (apparently less than 0.01 g/kg of the body weight) boost blood glucose levels slightly more slowly than corresponding doses of pure glucose. It was also found that the trehalose gives a slightly smaller, but still substantial, insulin response than the equivalent amount of glucose. There is no suggestion in this reference to use trehalose in diabetic foodstuffs, and indeed the teaching of the

reference suggests that trehalose should be used to achieve a quick boost in blood glucose levels. The reduction in insulin response of trehalose relative to glucose reported in this reference is insubstantial, and would not lead the skilled person to consider trehalose as a diabetic foodstuff.

EP-A-0693558 and EP-A-0850947 describe sweetener compositions comprising crystalline trehalose in combination with an intense sweetener such as ASPARTAME (Registered Trade Mark). The sweetener compositions are said to be useful as low-calorie sweeteners to sweeten foods for persons with diabetes or obesity. This can only be the case if the sweetener is used to replace a larger amount of sucrose in order to achieve a desired sweetness in a product.

(Trehalose itself is not a low calorie sweetener, since it is metabolised in the body and is less sweet than sucrose). In other words, these sweetener compositions must be substantially more sweet than sucrose, and must be used in substantially smaller amounts than sucrose, in order to achieve the stated advantages.

CN-A-1154214 describes proteinaceous foodstuffs containing 36-42% of protein, 8-10% of dietary fiber and 5 to 8% by weight of trehalose. The foodstuffs are said to be useful as diabetic foods. Consumption of normal amounts of these foodstuffs would result in relatively low doses of trehalose. Furthermore, this level of dietary fiber would be expected to reduce significantly the digestion of trehalose.

The present inventors have found that larger doses of orally administered trehalose behave very differently from the small doses described in the above documents. Larger doses of trehalose are absorbed more slowly, and provoke a very much smaller insulin response in healthy subjects than corresponding oral doses of glucose. The reasons for this behaviour are not clear, but it may be due to a limiting rate for trehalose breakdown and transport in the small intestine. In any case, it leads to the surprising conclusion that trehalose may be especially suitable for the nutrition of diabetics and other persons suffering from disorders of insulin metabolism.

Accordingly, the present invention provides a method of nutrition of a person suffering from a disorder of insulin metabolism comprising the step of oral administration of a composition comprising trehalose in an amount of at least 0.1 grams of trehalose per kilogram body weight of said person.

The above finding also leads to the conclusion that the nutritional use of trehalose may be especially suitable for treatment or prevention of medical conditions mediated by insulin metabolism.

Accordingly, the present invention also provides the use of trehalose for the preparation of an edible composition for use in a method of treatment or prevention of disorders due to defective insulin metabolism comprising administration of the edible composition according to the method of the present invention.

The treatment referred to is not primarily curative, but rather is directed to reduction of the adverse effects that can arise from conventional nutrition of a person suffering from such a disorder by substitution of the trehalose composition for e. g a sucrose composition.

The preventive use of trehalose according to the present invention is envisaged because it is thought that the use of trehalose in nutrition instead of conventional sugars such as sucrose may reduce the incidence of disorders of insulin metabolism in persons predisposed to such disorders. Such preventive use of trehalose is more a matter of dietary management performed by healthy individuals rather than active medical prophylaxis.

The disorder mediated by insulin metabolism may, for example, be diabetes (Type I or Type II) or hyperinsulinaemia. Conditions leading to increased risk of defects in insulin metabolism include obesity, heart problems, stroke, or physical trauma or disease of organs involved in insulin metabolism. Preferably, the disorder includes diabetes. The use of trehalose according to the present invention may be especially useful for the prophylaxis of Type I I diabetes.

Preferably, the nutritional composition comprises at least 10% by weight of trehalose, more preferably at least 20% by weight of trehalose, still more preferably at least 30% by weight, and most preferably at least 40 or 50% by weight of trehalose based on the total weight of dry substance in the composition.

Preferably, the composition comprises less than 8% by weight of dietary fiber, more preferably less than 5% by weight of dietary fiber, and most preferably less than 2% by weight of dietary fiber.

Preferably, the composition is selected from the group consisting of chocolate, hard sweets, biscuits, fondants, jellies, jams, sauces, puddings, syrups, soft drinks, sweet or savoury snack foods, cakes and other baked goods, ice cream, and combinations thereof. The composition may further comprise an intense sweetener such as saccharin or aspartame to bring the sweetness of the composition up to the level of an equivalent composition made with sucrose.

However, the total sweeteners in the composition preerably have a combined sweetness less than 1.5 times the sweetness of sucrose, more preferably less than 1.2 times the sweetness of sucrose. This enables the sweeteners, consisting mainly of trehalose, to fulfil the calorific and other (e. g. bulking and humectant) roles of sucrose in conventional foodstuffs.

Trehalose has been found to produce more acceptable diabetic or dietetic food products, for example with improved organoleptic properties or the lack of unpleasant gastro-intestinal effects associated with some carbohydrates such as fructose or sorbitol commonly used in such foods.

The step of oral administration comprises administration of at least 0.1g of trehalose per kg body weight of the diabetic person, preferably at least 0.3 g/kg, more preferably at least 0.5g/kg, and most preferably at least 0.7 g/kg. The benefits of trehalose use in accordance with the present invention are especially notable at the higher doses.

The present invention also provides a diabetic packaged food product or beverage for individual consumption comprising at least 10 grams of trehalose.

The food product is portioned, and preferably individually packaged, for consumption of the whole food product portion by one person substantially at one time. Examples include confectionery products such as candy bars, individual ice creams, individual baked products such as individual cakes, individual soft drinks such as cans of carbonated beverages, or individual desserts such as individual puddings, jellies, mousses or milk based desserts.

Preferably, the individual food product contains at least 20g of trehalose, more preferably at least 30g of trehalose, still more preferably at least 40 g of trehalose, and most preferably at least 50g of trehalose. Preferably, the individual food product weighs from 20 to 250 grams, more preferably from 40 to 150 grams.

Preferably, the individual beverage volume is from 150 to 1000 ml, more preferably from 200 to 750 ml. Candy bars weighing 40 to 100g and soft drink cartons, cans or bottles having volumes of 250 to 550mut are especially preferred.

Preferably, the weight concentration of trehalose in the food product is at least 25% based on the weight of the food product excluding packaging. Preferably, the concentration of trehalose in the beverage is at least 100mg/ml.

The trehalose in the packaged food products according to this aspect of the invention preferably substantially replaces the sucrose and other sugars such as maltose and glucose that are unsuitable for consumption by diabetics. Preferably, the food product or beverage comprises less sucrose than trehalose, and more preferably the food product comprises less than 10 g of sucrose, glucose or maltose. Most preferably, the food product comprises less than 10% by weight of sucrose plus glucose plus maltose, and the beverage comprises less than 50 mg/ml of sucrose plus glucose plus maltose.

The packaged food products or beverages according to this aspect of the invention may contain added sweeteners such as ASPARTAME (Registered Trade Mark). However, preferably the food products or beverages are not low

calorie products. Preferably, the food products have a calorie content of at least 1 kcal/g, more preferably at least 2 kcal/g, and still more preferably at least 3 kcal/g.

Preferably, the beverages have a calorie content of at least 0.2 kcal/ml, more preferably at least 0.3 kcal/ml and most preferably at least 0.4 kcal/ml.

Specific embodiments of the present invention will now be described further with reference to the following drawings, in which: Figure 1 shows the effect of ingesting 5% and 10% aqueous trehalose solutions on plasma glucose concentration of healthy (non-diabetic) subjects.

Data are also shown for a placebo and for a comparative 5% glucose aqueous solution; and Figure 2 shows the effect of ingesting the same beverages as in Figure 1 on plasma insulin concentration of healthy (non-diabetic) subjects.

Example 1 Eight healthy male human volunteers, aged 21 1 years (mean S. E. M.), body mass 74.0 7.4 kg, height 1.79 0.02 m, body mass index (BMI) 24.0 1.0 k/m2 volunteered as subjects for the experiment and gave written informed consent. None of the subjects had suffered an illness in the preceding 3 weeks.

None of the subjects had recently modified their dietary energy intake and none had undergone marked weight changes in the previous 3 months. Subjects were required to abstain from alcohol intake in the 24 hours preceding each experimental trial. All subjects were given a test drink containing 5 g trehalose prior to commencing the main study. None of the subjects reported any gastrointestinal discomfort following the test drink.

Subjects arrived in the laboratory between 9.00 and 11.00 a. m. following an overnight fast. A 4 cm, 21 g Venflon (registered trade mark) cannula was inserted into an antecubital vein. On four separate occasions subjects consumed 0.7 g per kg body mass of glucose, trehalose or placebo solutions (each solution had the same lemon flavour and contained 20 mmol/I trisodium citrate). Subjects were

asked to consume the drinks within a 3 minute period. The glucose solution contained 5% w/v glucose and two trehalose solutions were used containing 5 and 10% w/v trehalose. The placebo contained saccharin as sweetener but no carbohydrate. Hence, the mean fluid intakes were 1036 104 ml for the placebo (PLA), glucose (GLU) and 5% trehalose (T5) drinks and 518 52 ml for the 10% trehalose (T10) drink. The mean sugar intakes were 51.8 5.2 g for the glucose and trehalose treatments. The osmolality of the drinks was measured using a freezing point osmometer (Advanced Instruments) and was 110,361,255 and 386 mOsm/kg for the PLA, GLU, T5 and T10 drinks, respectively (note that 80 mOsm/kg of each drink was attributable to the added trisodium citrate). The order of the treatments was randomised and blinded to the subjects. The subjects remained seated throughout the 3 hours. During each test the subjects were asked about the palatability of the drinks and they were also asked to report any gastrointestinal discomfort experienced during and following the tests.

Blood samples were obtained before consumption of the drink and at 5,10 and 15 minutes after consumption of the drink. Further blood samples were obtained at 15 minute intervals for up to 3 hours after consumption of the drink.

About 5 mi of blood was placed into heparinised tubes. An aliquot was used to determine haemoglobin concentration and haematocrit so that plasma volume changes could be estimated according to Dill and Costill J. Appl. Physiol. Vol 37, pages 247-248 (1974). The remainder of the blood was centrifuged at 1500 g at 4°C to obtain plasma. The latter was stored at-70°C prior to analysis for glucose using an enzymatic, spectrophotometric method (Sigma Chemicals kit HK-20), insulin (by radioimmunoassay using an ICN Biomedicals insulin antibody coated tube kit) and total protein (Biuret method, with bovine serum albumin as standard) concentrations. The coefficient of variation for the assays was 1.9% for glucose, 0.5% for protein and 3.0% for insulin.

Differences with time and treatment were assessed using a repeated measures ANOVA with Tukey post hoc tests where appropriate. The accepted level of significance was P<0.05. All data in the text and Figures are reported as

mean standard error of the mean (S. E. M.) for eight subjects from whom we obtained the full complement of blood samples.

None of the subjects experienced any gastrointestinal discomfort during or following the tests. Subjects reported that the drinks were palatale.

The mean resting plasma glucose concentration before consumption of the drinks was 5.1 mmo1/1. The change in plasma glucose concentration following consumption of the different drinks is shown in Figure 1. On the PLA treatment the plasma glucose concentration remained stable throughout the 3 hour period.

Following GLU ingestion, plasma glucose concentration rose significantly after 10 minutes and reached a peak of 7.64 0.57 mmo1/l alter 32 2 minutes and had returned to normal by 45-60 minutes. After 90-120 minutes the plasma glucose concentration fell to a minimum of 3.59 0.21 mmol/l; at this point the plasma glucose concentration was significantly lower than that observed on the T5 trial.

With the T5 drink, the plasma glucose concentration was significantly higher than on the PLA trial only 5 minutes after consuming the drink. Plasma glucose concentration on the T10 trial was not significantly greater than PLA until 15 minutes alter consuming the drink, but the plasma glucose concentration at this time was still significantly lower than on the GLU trial. After 30 minutes plasma glucose levels began to fall on both the GLU and T5 trials and at 45 minutes were not significantly different from PLA. On the TIO trial the plasma glucose concentration was higher than PLA after 45 minutes. The area under the curve for the first 60 minutes (AUC60) following consumption of the drinks was not significantly different for GLU compared with either trehalose drink (P<0.05).

There was, however, quite a large difference between individuals (range 23-97%).

As a percentage of the GLU-AUC, the mean values for T5 and T10 were 55 17% and 84 17%, respectively.

Changes in plasma volume were minimal and nonsignificant following all of the drinks and there were no significant differences between the treatments. At the end of the 3 hour period, changes in plasma volume were +3.7 3.2%, +2.2

1.9% +4.0 2.6% and +3.7 3.2% for the PLA, GLU, T5 and T10 trials, respectively.

Plasma protein concentration fell on average by 7% during the tests (P<0.01) but there were no significant differences between the treatments.

The mean resting plasma insulin concentration before consumption of the drinks was 18 mU/I. The change in plasma insulin concentration following consumption of the different drinks is shown in Figure 2. On the PLA treatment the plasma insulin concentration remained stable throughout the 3 hour period.

Following GLU ingestion, plasma insulin concentration increased and reached a peak of 92 31 mU/I after 30 minutes and had returned to normal by 90-120 minutes.

With both trehalose drinks, the plasma insulin concentration at 15 and 30 minutes was significantly lower than on the GLU trial (P<0.05) and the peak plasma insulin values observed during the each of the trehalose trials were significantly less than that on the GLU trial (P<0.05; Figure 2). The time to reach peak insulin concentration was longer for the trehalose drinks compared with GLU.

The area under the curve for the first 60 minutes (AUC60) following consumption of the drinks was higher for GLU compared with either trehalose drink. As a percentage of the GLU-AUC, the mean values for T5 and T10 were 28% and 30%, respectively.

The results of the study show that trehalose has a glycaemic index that is on average a little lower that that of glucose, but is still medium-high. There was, however, a wide variation between different individuals. Those subjects who had a large AUC60 on T5 also showed a large response on T10, which suggests that interindividual differences in gut trehalase activity may be responsible for this large degree of variation. Plasma glucose concentration was maintained at or above basal (placebo) levels for longer with T10 compared with GLU, and there was no significant reactive hypoglycaemia with T5 or T10, whereas this was noticeable 90-120 minutes after GLU ingestion. This is likely to be due to the substantially

larger peak insulin response following GLU ingestion compared with the trehalose drinks.

Example 2 Foodstuffs in accordance with the present invention are prepared as follows: (a) Strawberry Flavour Hard Candy.

The candy was formulated as follows: % Trehalose 78.7 Isomalt 19.7 Anhydrous Citric acid 1.0 Strawberry Flavour D4 888 0.3 Strawberry Colour Hexacol 73234 0.3 The hard candy was prepared in accordance with the following instructions: 1. Gently heat trehalose, isomalt and sufficient water to dissolve the sugars.

2. Heat whilst stirring until dissolved.

3. Heatto 180°C.

4. Remove from heat and allow to cool to 130-140°C.

5. Add acid, flavour and colour with stirring.

6. Pour into moulds or stamp using conventional equipment.

(b) Milk Chocolat A milk chocolate was formulated as follows:- % Trehalose 47 Full cream milk powder (26.5% fat) 20.5 Cocoa butter 17.5

Cocoa mass 12.5 Butter oil 2.0 Lecithin (Topcithin 200) 0.5 Vanillin crystals 0.028 The chocolate was made with conventional chocolate processing machinery in accordance with the following instructions: 1. The trehalose was milled to 50pm.

2. Kneading-Add all the ingredients except the lecithin and enough cocoa butter to provide a suitable consistency for the roller refining operation.

Knead for 10 minutes at 40°C.

3. Refining-Standard 5 roll refiners cooled to remove any frictional heat. A single refining operation to give a suitable particle size (25-28, um).

4. Conching-as for making standard milk chocolate. The equipment is set to give a product temperature of 54°C. The remainder of the cocoa butter is added. The material is conched for a minimum of 4 hours.

5. Half an hour before the end of the conching period the emulsifier is added.

6. The chocolate was tempered with the same conditions as for standard milk chocolate (40°C-27°C-30°C) then moulded up.

(c) Tomato Ketchup A tomato ketchup was formulated as follows:- % Concentrated tomato puree 27.0 Water 24.42 Trehalose 25.0 Distille malt vinegar 20.0 Starch 2.0 Potassium Chloride 1.28 Salt 0.6 Tomato Ketchup Spice LID-SM986 0.2

The tomato ketchup was prepared in accordance with the following instructions:- 1. Weigh out all ingredients except water and starch into pan.

2. Disperse starch in water then add to pan.

3. Heat to 85°C for 5 minutes.

4. Blend with Silverson at maximum setting for 10 minutes.

5. Heat to 85°C for 5 minutes.

6. Hot fill into sterilised jars.

(d) Digestive Biscuits The biscuits were formulated as follows:- % White biscuit flour 52.8 Vegetable fat 18.7 Skimmed milk powder 0.6 Water 6.3 Sodium bicarbonate 0.2 Ammonium bicarbonate 0.1 Trehalose 20.7 Salt 0.6 The biscuits were prepared in accordance with the following instructions:- 1. Mix fat and sugar.

2. Dissolve salt in the water an 3. Mix.

4. Mix the flour, sodium bicarbonate and ammonium bicarbonate separately.

5. Combine with rest of mixture.

6. Rest dough is sealed bag for one hour at room temperature.

7. Process dough into biscuit shapes.

8. Bake on biscuit wire at 225°C for 6 minutes.

(e) Sponge Cake

The sponge cake was formulated as follows:- % Flour 32.0 Trehalose 22.2 Skimmed milk powder 2.2 Water 14.7 Baking powder 2.1 Past frozen egg 24.4 Emulsifier 2.1 Salt 0.3 The cake was prepared in accordance with the following instructions:- 1. Weigh egg, pettina whip, and water into Hobart mixing bowl.

2. Sieve dry ingredients together and add to bowl.

3. Mix. speed 1-1 min (scrape bowl) speed 2-1 min (scrape bowl) speed 3-2.5 min 4. Check batter density is 0.4 g/ml.

5. Weigh 200 g of mixture into tin.

6. Bake for 20 minutes at 170°C.

The above embodiments have been described by way of example, only.

Many other embodiments falling within the scope of the present invention will be apparent to the skilled reader.