FORMULATIONS OF ALPHA-AMYLASE INHIBITORS WITH ALPHA-GLUCOSIDASE INHIBITORS USEFUL IN THE TREATMENT OF DIABETES AND OBESITY

Field of invention

The present invention relates to formulations consisting of (i) Phaseolus vulgaris extracts containing pharmacologically active quantities of α-amylase inhibitors and (ii) Salacia oblonga or Salacia reticulata extracts containing pharmacologically active quantities of α-glucosidase inhibitors.

Background to the invention

Obesity is currently one of the major health problems, especially in the industrialised countries, with serious consequences in cardiocirculatory and skeletal terms. Carbohydrates are an important source of calories, and contribute to the synthesis of fats in individuals predisposed to obesity or type II diabetes. As hyperglycaemia leads to an increase in energy deposits, the availability of substances that reduce bioavailable glucose is very important. As starches are the main source of glucose, specific α-glucosidase and α-amylase inhibitors, obtained from plant materials or by synthesis, have been studied. It has long been known that some seeds and pulses contain substances which can have adverse effects on the diet if eaten before they are completely cooked. Many pulses contain protease inhibitors, amylase inhibitors and substances that discourage predators from continuing to use them by reducing the appetite. These substances, called phytohaemagglutinins, can cause hyperplasia of the pancreas at high doses, but can be useful in appetite control at lower doses.

At high doses, these lectins survive the intestinal transit and bond to the enterocytes where they cause the secretion of cholecystokinin, a trophic

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hormone that stimulates secretion by the pancreas, consequently causing its enlargement. Cholecystokinin also has favourable effects, because it reduces the appetite by reducing gastric motility.

As excess blood glucose leads to an increase in energy deposits, substances that reduce the availability of free glucose after an intake of starchy carbohydrates in the diet are very important. Most glucose originates from the breakdown of starch, which begins in the mouth due to the effect of ptyalin; this enzyme detaches saccharide chains, which in turn are converted to glucose by saccharases and α-glucosidases. α-amylase, secreted by the pancreas in the duodenum, demolishes the starch in glucose chains in the intestine, where it is converted to glucose due to the effect of α-glucosidase.

It is therefore obvious that in order to achieve a substantial reduction in the release of glucose, it is important to have both α-amylase inhibitors and α-glucosidase inhibitors.

The α-amylase inhibitor in itself considerably reduces the quantity of glucose originating from the starches present in the diet, and reduces the appetite after repeated administration.

It has now surprisingly been found that by combining Phaseolus vulgaris extracts with Salacia oblonga extracts in the ratio of between 1 : 1 and

1 :4, a marked reduction in blood glucose is obtained in humans. It is also known that a reduced food intake leads in any event to a reduction in the accumulation of fatty deposits.

Description of the invention It has surprisingly been found that by preparing products enriched with α-amylase inhibitors having a lectin content that ensures excellent tolerability, a reduction in body weight proportional to the dose administered can be obtained. The data in rats suggested that the effect on body weight reduction

was not simply associated with a reduction in the plasma glucose level, but also with a definite reduction in food consumption. Various pharmacological experiments demonstrated that this reduced food intake, despite unrestricted access to food, was associated not with a simply toxic effect, but with a modification in the desire to eat. The Phaseolus vulgaris extract described in PCT/EP2006/012012 is preferably used. Said extract is obtainable by extraction from Phaseolus sp. with mixtures of ethanol and water, and is characterised by an alpha-amylase inhibitor content of between 1200 and 1600 USP/mg (HPLC titre between 7 and 14% w/w) and a phytohaemagglutinin content of between 12,000 and 30,000 HAU/g. Said extract can be obtained by a process which comprises: a) extraction from Phaseolus sp. with aqueous buffers having a pH varying between 3 and 6.5 and subsequent separation of the extract from the biomass, which can possibly be further extracted with the buffer until exhaustion in alpha-amylase and phytohaemagglutinin inhibitors; b) filtration or centrifugation of the combined extracts and concentration to a volume corresponding to approx. 10% of the weight of the biomass of the initial extract after centrifugation; c) differential precipitation of the concentrated aqueous extract with diluted ethanol, to a final concentration of between 60 and 70% v/v; d) separation of the precipitate and reprecipitation from demineralised water with 60% ethanol, or diafiltration on a membrane with a 10,000 Da cut-off and drying of the precipitation residue.

The combination of an extract with α-amylase inhibitors and an extract containing α-glucosidase inhibitors, such as extracts of Salacia oblonga or

Salacia reticulata root or similar commercially obtainable species, has surprisingly demonstrated an unforeseen synergic effect, with a reduction in plasma glucose levels not obtainable with the individual ingredients.

The results for the hypoglycaemic activity of the combination and the individual extracts are set out in the Table below according to the method described by Tormo MA et al., Br. J. Nutr. 96, 539, 2006.

Table

Effect of Phaseolus vulgaris extract, Salacia oblonga extract and their combination on glycemia in Wistar rats given a restricted amount of food with a 1 hour/day limited access

Number of animal/group: 8 ^' * p<0.05 ** p<0.01 vs controls

The administration of the individual compounds and the corresponding combination did not have any toxic effect at doses of up to 2000 mg/kg for three months.

The present invention allows the preparation of products useful in the treatment of obesity, excess weight and type II diabetes, and in diets designed to maintain a constant body weight.

In order to be effective on blood glucose and to be effective on weight control in long-term treatments, the two extracts are combined in a ratio of between 1 : 1 and 1 :4; the doses range from 50 to 200 mg for the Phaseolus vulgaris extract and from 200 to 600 mg for the Salacia extract, depending on the active constituent content measured by HPLC. The formulations according to the invention can take the form of soft or hard gelatin capsules, cellulose capsules, tablets or liquid forms, and the extracts can be separately formulated in totally or partly gastroresistant forms. The products should be taken at main

meals or whenever foods rich in carbohydrates are eaten. The formulation can also be associated with substances that reduce gas formation in the colon or partly absorb the excess.

The following examples illustrate the invention in detail. Example 1 - Formulation of Phaseolus vulgaris and Salacia oblonga extracts into hard gelatin capsules Unit composition:

Phaseolus vulgaris dried extract 50 mg

Salacia oblonga dried extract 200 mg Microcrystalline cellulose 120 mg

Mannitol 100 mg

Simeticone 20 mg

Silicon dioxide 10 mg

Manufacturing process: - Adsorb the simeticone on the mannitol and on part of the microcrystalline cellulose.

Mix the resulting product with Phaseolus vulgaris dried extract, Salacia oblonga dried extract, the remainder of the microcrystalline cellulose and the silicon dioxide. - Divide the mixture between hard gelatin capsules.

Example 2 - Formulation of Phaseolus vulgaris and Salacia oblonga extracts into tablets

Unit composition:

Phaseolus vulgaris dried extract 100 mg Salacia oblonga dried extract 150 mg

Microcrystalline cellulose 150 mg

Mannitol 100 mg

Cross-linked sodium carboxymethylcellulose 30 mg

Silicon dioxide 8 mg

Magnesium stearate 5 mg

Manufacturing process:

Mix the Phaseolus vulgaris dried extract, Salacia oblonga dried extract, microcrystalline cellulose, mannitol, crosslinked sodium carboxymethylcellulose and silicon dioxide for 10 minutes.

Add the magnesium stearate to the mixture and proceed with mixing for a further 2 minutes.

Compress the mixture obtained in the preceding point. Example 3 - Formulation of Phaseolus vulgaris and Salacia oblonga extracts into modified-release granules Unit composition:

Phaseolus vulgaris extract 50 mg

Salacia oblonga extract 100 mg Microcrystalline cellulose 100 mg

Povidone 10 mg

Sodium carboxymethylcellulose 8 mg

Methacrylic acid copolymer 50 mg

Triethyl citrate 3.2 mg Talc 8 mg

Simeticone 0.3 mg

Manufacturing process:

Granulate the extracts, microcrystalline cellulose and sodium carboxymethylcellulose with an aqueous solution of povidone. - Dry and calibrate the granulate obtained.

Coat the granules with an aqueous suspension of methacrylic acid copolymer, triethyl citrate, talc and simeticone.

Example 4 - Formulation of Phaseolus vulgaris and Salacia oblonga

extracts into immediate-release granules

Unit composition:

Phaseolus vulgaris extract 50 mg

Salacia oblonga extract 100 mg Microcrystalline cellulose 100 mg

Povidone 10 mg

Sodium carboxymethylcellulose 8 mg

Manufacturing process:

Granulate the extracts, microcrystalline cellulose and sodium carboxymethylcellulose with an aqueous solution of povidone.

Dry and calibrate the granulate obtained.

Example 5 - Mixture of granulates of Phaseolus vulgaris and Salacia oblonga extracts with different release profiles

Manufacturing process: - Mix 50% of the granulate described in example 3 with 50% of the granulate described in example 4.

Divide the mixture obtained between hard gelatin capsules.