IE57349B1 - Compositions comprising peptides - Google Patents

Abstract

1. Compositions containing glucides, vitamins, mineral salts, a lipid fraction and a protein fraction, characterized in that the protein fraction contains the following 3 types of minipeptides : - lactoserum minipeptides, - total casein minipeptides, - casein minipeptides without phosphopeptides.

Description

The present invention relates to new compositions tor dietetic, reanimation or therapeutic use, containing a protein traction based on 3 types of minipeptides.

Numerous products based on minipeptides for 5 use in dietetics or therapy and more particularly, in therapeutic nutrition, are already on the market.

These products possess a protein traction containing, in varying proportions, free amino acids and minipeptides with various lengths of chain. A few years ago, there were only products based on amino acids but in recent times, products based on small peptides have become more preferred. Thus products based on amino acids are always hyper-osmolar and hence often lead to digestive intolerances. Clinical research, like for example, that of SILK D., PERRETT D., CLARK M., Intestinal transport of two dipeptides containing the same two neutral amino acids in man, Clin. Sci. Mol Med 45: 291-299, 1973, have shown, moreover, the improved absorption of small peptides as compared with free amino acids.

In addition, the works of KEOHANE P., BROWN Barbara, GRIMBLE G., SILK D., The peptide nitrogen source of elemental diets - comparisons of absorptive properties of five partial enzymic hydrolysates of whole protein, A.S.P.E.N. 6th Clinical Congress, 3-6 February 1982, San Francisco, showed the improved absorption of small peptides when the latter « result from a mixture containing, among others, 1 casein peptides and lactoserum peptides, as compared * with peptides of other origin.

However, a protein mixture based solely on casein minipeptides and lactoserum minipeptides in a composition intended for dietetic, reanimation or therapeutic use would give rise either to a too high level of sulphur-containing amino acids, if the lactoserum peptides were in too high quantity; or to a too high level of phosphorus bound to protein if on the other hand the casein minipeptides were in too high a quantity.

It was therefore desirable to produce a mixture of casein minipeptides and lactoserum minipeptides which while retaining the advantageous adsorption properties of these 2 types of minipeptides still ensured appropriate levels of both organic phosphorus bound to protein and amino acids containing sulphur.

Only casein minipeptides contain phosphorus 10 bound to protein; lactoserum minipeptides do not contain any. The necessary balance between casein minipeptides and lactoserum minipeptides leads to an excessive contribution of phosphorus bond to protein. We have therefore developed a composition in which a part of the total casein minipeptides is replaced by casein minipeptides without phosphopeptide In this way it is possible to diminish in these compositions the level of phosphorus bound to proteins resulting from the casein while retaining the desired level of sulphur-containing amino acids.

According to the present invention we provide compositions comprising glucides, vitamins, mineral salts, a lipid fraction and a protein fraction, the said protein fraction containing lactoserum minipeptides, total casein minipeptides and casein minipeptides without phosphopeptides.

The lactoserum minipeptides for use in the compositions according to the invention may be obtined by total enzyme hydrolysis of lactoserum proteins. This enzyme hydrolysis process and the minipeptides obtained by the same process are described in European Patent Specification No. 0022019.

The two other types of minipeptides contained in the protein fraction of the compositions of the invention may be prepared starting with phosphocaseinates of monovalent or bivalent cations, treated according to the processs indicated in the two European patents published under numbers 0033686 and 0034083.

These two patents not only indicate how the total casein minipeptides may be obtained by enzyme hydrolysis of the phosphocaseinates of monovalent or bivalent cations but also describe the different stages which enable, for the one part phosphopeptides and for the other part minipeptides without phosphopeptides to be separated, starting with total casein minipeptides.

A particular advantage of the compositions according to the invention is that they contain a protein fraction which consists essentially of minipeptides and is practically devoid of free amino acids (present in a proportion of less than 15%). The compositions according to the invention also have the advantage of containing minipeptides which by their nature are very well absorbed by an organism.

The compositions according to the invention are of particular use for invalids and may, if required, form a part of therapy. Thus, for example, the contribution of sulphur-containing amino acids and the contribution of phosphorus bound to proteins determines the charge in H+ ions, an important parameter in invalids. Very often illnesses of insufficiency of kidney function give rise to inadequate regulation of the acid-base balance and in such cases a strict control of the H+ ion intake is essential. By adjustment of the level of organic phosphorus in the compositions according to the invention the charge of h ions in the compositions may be readily varied.

The present invention provides compositions in which the associated three types of minipeptides enable variation of the level of phosphorus bound to proteins. The level of phosphorus bound to proteins in these compositions depends on the proportion of the phosphoprotein part contained in these compositions. In order to obtain a physiological level of organic phosphorus, the phosphoprotein fraction should represent, preferably, from 1% to 10% by weight of the compositions of the invention. According to a preferred embodiment of the invention, by appropriate combination of the 3 types of minipeptides, compositions may be obtained in which the level of phosphorus bound to proteins is at least equal to that present in human milk, that is to say of the order of 1.5 mg per g of protein and at most is equal to that present in cows' milk, that is to say of the order of 6.6 mg per g of protein.

The present invention therefore has as a particular feature compositions wherein the weight of the protein fraction represents 1% to 10% of the total weight, and especially those wherein the level of phosphorus bound to protein is from 1.5 mg per g of protein to 6.6 mg per g of protein.

Furthermore the distribution of amino acids in the compositions of the invention is variable depen20 ding on the respective quantities of the 3 types of minipeptides. Thus, the 3 types of associated minipeptides permit different aminograms to be obtained, e.g. adapted to specific pathological situations.

A combination of the 3 types of minipeptides, in approximately equal quantities, in the protein fraction of the compositions according to the invention, enables a distribution of amino acids to be available which ensures maximum nutritional efficiency.

Under these conditions, the level of phosphorus bound to protein is about 3 mg/g of protein. Particular compositions of this type which may be mentioned are those wherein the distribution of amino acids of the protein fraction is that given below, the following figures being expressed in grams of amino acid per 100 g of amino acids: He 5.1 Val 5.9 Leu 10.1 Arg 3.3 Lys 8.5 His 2.5 Met 2.3 Ala 3.6 Cys 1.6 Asp 8.4 Phe 4.6 Glu 15.9 • Tyr 4.9 Gly 1.8 Thr 4.4 Pro 8.4 1 Trp 1.8 Ser 4.7 This aminogram permits a high ratio of branched amino acids, while retaining a high level of lysine.

The compositions of the invention may provide all the other necessary nutrients in addition to the protein element, in appropriate proportions.

Thus they contain glueides, ensuring 50% to 60% of the total energy contribution (=T.E.C.), and vitamins (e.g. A, D, E, C, B^, B2, PP, Βθ, B^r folic acid, biotin, B^f K^, choline) .

The compositions according to the invention can satisfy the physiological need for mineral elements. They also contain a lipid fraction which may provide 30% - 35% of the T.E.C.

The invention has notably as its subject compositions in which the lipid fraction has approximately the following composition: - medium chain triglycerides 56.4% - Oenothera oil 25.6% - soya oil 18 % More than half of this lipid fraction is composed of short or medium chain triglycerides.

This gives use to good tolerance, even in invalids suffering from malabsorption or maldigestion of lipids or from a bad metabolization of essential fatty acids. Thus short or medium chain triglycerides are easily assimilable by the organism, even when digestion is disturbed. - 7 Such a lipid fraction ensures the supply of the following essential fatty acids: - linoleic acid (=9.6% of the T.E.C.) - y-linoleic acid (=0.8% of the T.E.C.) - α-linoleic acid (=0.4% of the T.E.C.).

The particular acids named above and/or their usual higher polyunsaturated derivatives may be added if desired. Such an addition may be carried out, for example, by adding synthesized triglycerides of 2 sorts: - synthesized triglycerides in position 1, 2 or 3 of linoleic or ^-linoleic acid (C18:3,&>6) , of dihomo-α-linoleic acid (C20:3,w6), of arachidonic acid (C20:4,u>6), of docosatetraenoic acid (C22:4,<»6) , or of docosapentaenoic acid (C22: 5,^6); - synthesized triglycerides in position 1,2 or of α-linoleic acid (C18:3,m3), of eicosatetraenoic acid (C20:4, As mentioned above the compositions of the invention are of particular use where strict control is necessary for example in therapy and reanimation. Thus the compositions may be used as food or food supplements, tailored for specific nutritional needs. The compositions of the invention may be used when post-operative nutritonal assistance is necessary. They are also of use in pre-operative nutrition, in particular when a residue-free diet is desired. Thus the compositions of the invention are completely absorbed in the proximal part of the small intestine, leaving only a minimal faecal residue.

The compositions of the invention may also be used in the case of disorders in the alimentary canal, and notably when the intestinal surface is reduced or display disease. This is the case for example with ailments characterized by a chronic inf lamination associated with fistulae, or by absorption trouble caused by a reduction of the surface of the small intestine. It is also the case with Crohn's disease, and chronic intestinal diseases.

The compositions of the invention also find use when problems in digestive secretion lead to ♦ poor digestion or malabsorption such as in the following: * - acute pancreatitis, - malabsorption syndrome caused by a malignant reticuclosis, - certain diseases of the system, - ischemic trouble of the intestine.

Furthermore, each time that there is an inflammatory attack in the intestine, a risk of allergization to whole proteins exists. The compositions of the invention may also be used to avoid the occurrence of allergies in the course of inflammatory diseases of the intestine.

The invention thus further relates to the compositions defined above in the form of food or food supplements as well in the form of therapeutic nutrition products. Such compositions may optionally being associated with a neutral vehicle suitable for oral or enteral administration.

The new compositions of the invention constitute, because of their properties, very useful medicaments in the treatment of the ailments described above.

The usual dose of the compositions of the invention varies depending on the condition of the patient, , the ailment in question and the administration route chosen. * For example, a dose of the composition of the invention may be administered ensuring a protein contribution of about 47 g to 118 g and preferably of about 71 g, a lipid contribution of about 58.5 g to 146 g and preferably of about 88 g, a glucide contribution of about 197 g to 493 g and preferably of about 296 g, it being understood that these doses are per day and by oral route in an adult.

A particular subject of the present invention is thus pharmaceutical preparations containing a composition according to the invention.

The compositions according to the invention may be presented in the forms currently employed in this field, either in liquid form or in the form of a powder. Thus, the compositions may be contained, for example, in metal boxes, flasks, bags or plastic bottles.

In the preparations given below in the examples, 1-5 g of maltodextrin may be replaced by galactose.

The examples given below illustrate the invention 15 without, however, limiting it.

Example 1 A liquid dietetic preparation was prepared for oral or enteral use, with the folllowing formulation: Centesimal composition - glueides - lipids — proteins - vitamins of RDA* and ESADDI** A, D, E, C, B^, ®2* 13.15 g (maltodextrins) 3.9 g consisting of: - Medium-chain triglycerides - Oenothera oil - soya oil 3.15 g consisting of: - lactoserum minipeptides - total casein minipeptides - casein minipeptides without phosphopeptide according to the American norms 2.2 g 1.0 g 0.7 g 1.05 g 1.05 g 1.05 g folic acid, biotin, 6’ 12’ g, K^, choline; - minerals : according to the American norms of RDA and ESADDI calcium, phosphorus, magnesium, iron, zinc, iodine, sodium, potassium, chloride, copper, manganese, selenium 0.404 g - vehicle g.s. for 100 ml *R.D.A. : Recommended Dietary Allowances 9th edition 1980 Committee on Dietary Allowances, Food and Nutrition Board Division of Biological Sciences, National Research Council, National Academy of Sciences, Washington, D.C., 1980 ** E.S.A.D.D.I. : Estimated Safe and Adequate Daily Dietary Intakes (Nutrition Reviews, Vol. 38, No. 8 August 1980, p 291) (Journal of The American Dietetic Association, Vol 76, No. 3, March 1980).

I For information only, the lipid fraction of this preparation ensures the following contribution of essential fatty acids: - linoleic acid : 1.1 g - ί-linoleic acid : 0.08 g - α-linoleic acid : 0.04 g.

I Example 2 : A liquid dietetic preparation for oral or enteral use was prepared, with the following formulation: Centesimal composition - glucides : 15.6 g (maltodextrins) - lipids : 5.2 g - proteins consisting of: - medium-chain triglycerides 2.9 g - Oenothera oil 1.2 g - soya oil 1.05 g - lecithin 0.05 g : 6 g consisting of: - lactoserum minipeptides 2 g - total casein minipeptides 2 g - casein minipeptides without phosphopeptides 2 g - vitamins : according to the American norms of RDA and ESADDI ;A, E, C, B·^, B2, PP, B&, Bjl2' acid, biotin, B^, choline - minerals : according to the American norms of RDA and ESADDI calcium, phosphorus, magnesium, sodium, potassium, chloride, iron, zinc, copper, manganese, iodine, selenium 0.562 g - vehicle q.s. for 100 ml The preparation of 7500 litres of the composition of Example 1 (the 7500 litres being then separated into metal boxes), is carried out in 3 stages as follows: 1st stage; Preparation of a lipid emulsion The material used is the following: - 2 x 2000 litres stainless steel tanks, equipped with: * an agitation system * a heating and cooling system * a vacuum apparatus * a centrifugal pump * * a high pressure pump 1500 litres of osmosis-treated water is introduced into one 2000 litre tank then heated under agitation.

When the temperature reaches 65°C the following are added: - 75 kg of Onagre oil - 37 kg 500 of soya lecithin - 97 kg 500 of medium-chain triglycerides - 37 kg 500 of glycerol monostearate.

Agitation is continued and after starting the cooling of the mixture, vitamins E, A and D (previously dissolved in 48 kg 750 of soya oil) are added.

When the temperature is about 20°C, the first transfer is started by the high presure pump.

The pressure is adjusted to 50 kg/cm at 2 the second stage of the pump and to 250 kg/cm at the first.

At the outlet, the product is collected in the second tank. .

One proceeds in this way with 4 transfers by the high pressure pump, passing alternatively * from one tank to the other.

A rapid check of the size of the liposomes of the emulsions may be carried out with a Coulter Counter. 2nd stage : Preparation of a starch phase The material used is the following: - an 8000 litre stainless steel tank equipped with: * a mechanism for agitation * a vacuum apparatus - a 500 litre stainles steel tank equipped with: * a mixing turbine - a plate exchanger provided with 3 circuits: * one for heating * one for cooling to 15°C * one for cooling to 5°C - 2 centrifugal pumps of 20ra3/hour.

The mixer being in a closed circuit with the tank, the pumps and the exchanger, 3700 litres of osmosis-treated water are added and heated to 85°C.

When this temperature is reached, 195 kg of dehydrated starch (1 to 4% according to the viscosity desired) is added by the mixer and the temperature is kept at 85°C for 30 minutes.

At the end of this period, the temperature is lowered to 50 °C by means of the cooling section of the exchanger then the following are added successively: - 98 kg of lactoserum minipeptides, - 92 kg of total casein minipeptides, - 100 kg 500 of casein minipeptides without phosphopeptides, - 860 kg of maltodextrin.

The temperature s lowered to 40°C and the following are introduced: - magnesium chloride, - potassium hydrogen phosphate, - calcium chloride, - sodium chloride, - choline chloride, - potassium iodide and sodium selenite.

The temperature of the mixture is lowered to 25°C and the following are added: - ascorbic acid, - iron sulphate - nicotinamide or vitamin PP, - zinc citrate, * - calcium pantothenate, - manganese sulphate, * - pyridoxine hydrochloride or vitamin Bg, - thiamin hydrochloride or vitamin B^, - copper sulphate, - vitamin B^2 1% on mannitol, - riboflavin or vitamin B2, - folic acid, - biotin. 3rd stage : preparation of the final mixture Whilst the starch phase is under agitation in the 8000 litre tank, the lipid emulsion prepared in the first stage is introduced by means of a centrifugal pump. After adjusting the level of the tank to 7450 litres with osmosis-treated water and the pH to 7.10 by means of a potassium hydroxide solution, checking the level of the tank and measuring the dry material contents in the tank, the final vacuum is made and nitrogen is added to the tank.

The final mixture obtained in this way is then sterilized by filtration, at 150°C for 8 seconds by a passage in a coil circuit.

The final product is then distributed between 375 ml metal boxes; the latter being closed aseptically 0 (the boxes and lids are previously sterilized by superheated steam). » Clinical study of the product of Example 1 invalids, aged from 15 to 53 years were observed over a period of 14 days in the course of gastro-enterological reanimation. These people displayed serious gastro-intestinal disorders with one or more complications (for example : acute chlolecystitis with oedematous pancreatitis; bullet wound leading to duodenal injury, an injury of the liver, an injury of the straight colon, acute pancreatitis with toxi-infectious syndrome).

The aim of this study was threefold: 1) To study the tolerance of the product; 2) To evaluate the absorption; 3) Assess the nutritional impact of the product. Throughout the duration of this study, the six patients received food by enteral route only through a feeding tube with a continuous flow, hours out of 24.

This food was composed exclusively of the product of Example 1 for all the patients, except for one for whom an addition of rice water had been made because of a severe toxi-infectious syndrome.

The following table indicates, for each patient: - the energy contribution per 24h in Kcal, - the volume of the product of Example 1 administered per 24 hours. - the daily contribution in nitrogen (g/24h.) corresponding to the administration of nutritional support.

Patient 1 2 3 4 5 6 Energy contr ibution Kcal/24 h. 4125 4125 4125 2300 3750 3750 30 Volume of product of Example 1 in ml. per 24 h. 4125 (11 4125 x 375 4125 ml.boxes) 2250 (6 ml. 3750 x 375 boxes) 3750 (10 x 375 ml. boxes) 35 Nitrogen contribution g/24 h. 22 22 22 12 20 20 1) Study of the tolerance of the product The following have been used as criteria of tolerance: - acceptability, - vomitting, - abdominal pains, a - diarrhoea, - flatulence. b According to these criteria, with a daily intake of 3750 to 4125 ml. of product, the patients 1, 2, 3, 5 and 6 displayed an excellent tolerance.

For patient 4, the criteria of tolerance defined above were also good, with a reduction in the intake to 2250 ml/day of the product. 2) Study of the absorption and the nutritional efficiency The nutritional value of the product of Example 1 was studied by means of the following results (see Table I): - The coefficient of absorption of minipeptides of the product of Example 1, even in the case of severe gastro-enterological disorders and a toxiinfection, was greater than 95% in all but one case. Moreover, in this group of patients suffering from severe gastro-enterological problems which, in certain instances, makes any enteral nutrition impossible, the coefficient of absorption is distinctly greater than the average coefficient of absorption of the usual patients under treatment. .

- The nitrogen balance appears positive from the second day, varying between +9.25 and +15g/24h, » except for patient 4 (+3g). This patient has a toxi-infection necessitating a reduction in intake (12 g of nitrogen/24 hours).

Thus, even with a nitrogen and calorie intake clearly reduced, the efficiency of the product enabled the maintenance of a positive nitrogen balance.

- Albumin : one of the principal criteria of protein metabolism is the level of plasma albumin. The level of albumin increased significantly in the course of treatment.

Likewise, weight increase rose significantly (2.5 kg) .

In this study, all the other metabolic parameters remained identical or increased within normal limits, or improved.

Finally, the clinical development is excellent for the 6 patients.

The nutritional criteria and the clinical condition are thus improved in a similar way to the nitrogen balance.

PATIENT AGE COEFFICIENT 1 OF ABSORPTION OF MINIPEPTIDES (N) ALBUMIN DO D14 1 52 96.5 ·% 31 33.4 2 51 98.1 % 30.8 39 3 47 98.7 % 36.5 43.7 4 44 95.5 % 38 45.9 5 15 86.5 % 33 43.6 6 16 97.5 % 41 48 WEIGHT DO D14 OTHER METABOLIC PARAMETERS All the parameters remained the same or rose within the normal limits or improved. The nitrogen balance was positive i from the 2nd day 71 75 ' idem 70 72 ί idem 64 58 idem 43 45 idem 70 idem CLINICAL RESULTS Excellent progress with recovery idem idem Ser ious toxi-infection controlled + recovery Excellent progress with recovery idem Conclusion The product of Example 1, utilised in this clinical test on patients displaying severe gastrointestinal disorders with one or more complications, displays an excellent nutritional efficiency which enables the following to be ensured: - a stabilization or improvement of the biological parameters within the normal limits; - a coefficient of absorption of minipeptides greater 10 than 95%; - a clear, positive nitrogen balance; - an improvement in the clinical condition.

Furthermore, this product displays a very 15 good tolerance.

Claims (13)

Claims:

1. Compositions comprising glucides, vitamins, minerals salts, a lipid fraction and a protein fraction the said protein fraction containing lactoserum minipeptides, total casein minipeptides and casein minipeptides without phosphopeptides.

2. Compositions according to claim 1 wherein the weight of the protein fraction constitues 1% to 10% of the total weight.

3. Compositions according to claim 1 or claim 2 wherein the level of phosphorus bound to protein is from 1.5 mg per g of protein to 6.6 mg per g of protein.

4. Compositions according to any one of claims 1 to 3 wherein the 3 types of minipeptides are present in approximately equal quantities by weight.

5. Compositions according to claim 4 wherein the level of phosphorus bound to protein is about 3 mg/g of protein.

6. Compositions according to claim 4, wherein the distribution of amino acids in the protein fraction is that given below, the following figures being expressed in grams of amino acid per 100 g of amino acids: He 5.1 Val 5.9 Leu 10.1 Arg 3.3 Lys 8.5 His 2.5 Met 2.3 Ala 3.6 Cys 1.6 Asp 8.4 > Phe 4.6 Glu 15.9 Tyr 4.9 Gly 1.8 » Thr 4.4 Pro 8.4 Trp 1.8 Ser 4.7 ,

7. Compositions according to any one of claims 1 to 6 wherein the lipid fraction has approximately the following composition: - medium chain triglycerides - Oenothera oil - soya oil 56.4% 25.6% 18 % .1 I -M .1 I -M

8. Compositions according to any preceding claim in a form suitable for oral or enteral administration.

9. Compositions according to any preceding claim in the form of a food, food supplement or a therapeutic nutrition product.

10. Compositions according to any preceding claim containing a neutral vehicle.

11. Pharmaceutical preparations containing a composition according to any preceding claim.

12. Compositions according to claim 1 substantially as herein described.

13. Compositions substantially as herein described in Example 1 or Example 2.