WO2003105609A1

WO2003105609A1 - Infant formula supplemented with phospholipids

Info

Publication number: WO2003105609A1
Application number: PCT/IL2002/000674
Authority: WO
Inventors: David Rutenberg
Original assignee: Lipogen Ltd
Current assignee: Lipogen Ltd
Priority date: 2002-06-16
Filing date: 2002-08-15
Publication date: 2003-12-24
Anticipated expiration: 2004-12-16
Also published as: IL150240A; EP1592313A1; AU2002326121A1; EP1592313A4; CN1523964A; IL150240A0; CN100393246C; JP2005519995A; US20040022922A1

Abstract

The present invention relates to pharmaceutical or nutritional or food compostion for feeding infants, which is more similar to human milk in a powder or fluid form, having an effect of addresseing the shortcomings of the presently known infant formulas by the inclusion of a phospholipid supplement which contains phosphatidylserine or salts thereof as one of the effective ingredients and comprising at least 1% (w/w) phosphatidyserine out of the total phospholipid content of the composition, wherein the phosphatidylserine has a structural fatty acid chain derived from at least one raw material lecithin selected from the group consisting of soy bean lecithin, rapeseed lecithin, or egg yolk lecithin, and which is produced by reaction with phospholipase-D.

Description

INFANT FORMULA SUPPLEMENTED WITH PHOSPHOLIPIDS

FIELD AND BACKGROUND OF THE INVENTION

The present invention relates to an infant formula supplemented with glycerophospholipids (hereinafter phospholipids) and more particularly, to a formula

supplemented with phosphatidylserine.

Breastfeeding, the natural feeding mode, has multiple beneficial effects on the

infant and is known to be the most suitable diet for infant's nutritional requirements.

Jensen RG. (Textbook of Gastroenterology and Nutrition in Infancy, Second

Edition, edited by E. Lebenthal. Raven Press, Ltd., pp. 157-208, New York 1989) reviewed measurements of the total lipid content of human milk and reports that

phospholipids are found in the human milk lipids at levels of about 20 to 40 mg/dl.

G. Harzer et al. reports that the phospholipid composition of human milk lipids (weight % of total phospholipids) in day 36 postpartum is

phosphatidylcholine (PC) 24.9%, phosphatidylethanolamine (PE) 27.7%, phosphatidylserine (PS) 9.3%, phosphatidylinositol (PI) 5.4%, sphingomyelin 32.4%

(G. Harzer et al., Am J Clin Nutr., Vol. 37, pp. 612-621, 1983).

Phospholipids are essential components of growing tissue in particular nerve

cells. In the latter, lack of phospholipids can cause serious impairment in development

and problems in overt functions. The source of phospholipids in the buildup of neuronal tissue, originates in part from intracellular synthesis and in part from the diet.

Therefore dietary supplementation of phospholipids in lactating infants could be critical for proper development (For comprehensive reviews, see: J. N. Hawthorne &

G. B. Ansell, Phospholipids 1982, Elsevier, Amsterdam, and M. Shinitzky, "Membrane Fluidity and Cellular Function", in Physiology of Membrane Fluidity,

Shinitzky M. Ed. Vol. I, pp. 1-53, 1984, CRC Press, Boca Raton, FL.).

In the past, only pig or bovine phosphatidylserine was available in the market.

However, due to the "mad cow disease" this source is generally unacceptable for usage with infant formulations today. In recent years a reliable and healthy source of

phosphatidylserine, which is produced by reaction of vegetal lecithin with

phospholipase-D is available in the market. A specific process of enzymatic

conversion is known and is described in, for example, the article by Eibl A. and

Kovatchev S. (Eibl A. and Kovatchev S. "Methods in Enzymology" Vol. 72,

pp. : 632- 639, 1981).

REFERENCES

Jensen RG. "Textbook of Gastroenterology and Nutrition in Infancy", Second Edition, edited by E. Lebenthal. Raven Press, Ltd., pp. 157-208, New York 1989.

G. Harzer et al., "Changing patterns of human milk lipids in the course of lactation and during the day", Am J ClinNutr., Vol. 37, pp. 612-621.

J. N. Hawthorne & G. B. Ansell, Phospholipids 1982, Elsevier, Amsterdam.

Shinitzky M. Ed. "Membrane Fluidity and Cellular Function in Physiology of

Membrane Fluidity", Vol. I, pp. 1-53, 1984, CRC Press, Boca Raton, FL.

Eibl A. and Kovatchev S. "Preparation of phospholipids analogs by phospholipase-D"

Methods in Enzymology" Vol. 72, pages : 632- 639, 1981.

SUMMARY OF THE INVENTION

-It is an object of the present invention to provide an infant formula which is

more similar to human milk in a powder or solution form, having an effect of addresseing the shortcomings of the presently known infant formulas by the inclusion

of a phospholipids supplement which contains phosphatidylserine as one of the

effective ingredients.

The infant formula of the present invention contains phosphatidylserine or the salt thereof as the effective ingredient, wherein the phosphatidylserine has a structural fatty acid chain derived from at least one raw material lecithin selected from the group

consisting of soy bean lecithin, rapeseed lecithin, or egg yolk lecithin.

According to further features in preferred embodiments of the invention

described below, the phospholipids include phosphatidylserine or the salt thereof as the effective ingredient, wherein the phosphatidylserine derived from at least one raw

material lecithin selected from the group consisting of soy bean lecithin, rapeseed

lecithin, or egg yolk lecithin, and which is produced by reaction with

phospholipase-D.

The aforementioned phosphatidylserine as the effective ingredient in accordance with the present invention is produced by the enzymatic reaction with phospholipase-D using as the substrate soy bean lecithin, rapeseed lecithin or egg yolk

lecithin. The process will now be illustrated. A raw material lecithin (namely,

phosphatidylcholine) selected from soy bean lecithin, rapeseed lecithin or egg yolk lecithin is subjected to the process of transphosphatidylation with phospholipase-D in

the presence of L-serine or D-serine or DL-serine and water, thereby substituting the

choline group with the serine group through the hydroxyl group, to produce the

rearranged phosphatidylserine.

^"Any commercially available soy bean lecithin, rapeseed lecithin or egg yolk

lecithin may be used, with no limitation, as the raw material. As phospholipase-D for

use in the process of enzymatic conversion, use may be made of for example those from cabbage and actinomyces, if they have an activity on lecithin or hydrogenated

lecithin or lysolecithin in the presence of L-serine or D-serine or DL-serine and water

to produce phosphatidylserine.

A specific process of enzymatic conversion is known and described in for

example the article by Eibl A. and Kovatchev S. "Preparation of phospholipids analogs by phospholipase-D." ("Methods in Enzymology" Vol. 72, pp. : 632- 639,

1981), so no detailed explanation is described herein.

According to another aspect of the present invention there is provided a

method of feeding an infant comprising the steps of mixing an infant formula powder

in water for obtaining a fluid including the nutritional components and the

phospholipids supplement and feeding the infant with the fluid containing nutritional components and a phospholipid supplement wherein the phospholipids include

phosphatidylserine or the salt thereof as the effective ingredient. According to still further features in the described preferred embodiments at

least some of the nutritional components are derived from milk or soy.

The present invention successfully addresses the shortcomings of the presently known configurations by providing an infant formula which is more similar to human

milk, adds protection from serious impairment in development and problems in overt

functions and improve the development and maturation of tissue in nerve cells.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present invention is of an infant formula supplemented with

phospholipids, particularly which includes phosphatidylserine, which can be used to

feed infants. Specifically, the present invention can be used to protect infants of

syndromes associated with feed devoid of phospholipids fed to them in the first year of their lives. The present invention renders infant formulas more similar to human milk. Thus, in accordance with one aspect of the present invention there is provided

an infant formula in a powder or fluid form which formula includes nutritional components and a phospholipid supplement, particularly which includes

phosphatidylserine.

In accordance with another aspect of the present invention there is provided a

method of feeding an infant. The method is effected by executing the following steps:

First an infant formula powder containing nutritional components and a phospholipid supplement is mixed in water for obtaining a mixture including the nutritional components and the phospholipid supplement. Second, the mixture is fed to the infant. As exemplified in the Examples section below, the nutritional components

may include milk or soy derived nutritional components. They may additionally

include one or more of the following ingredients: lactose, vegetable oils, skimmed

milk powder, whey protein concentrate, Sodium, Calcium, Phosphorus, Potassium,

Chloride, Iron, Magnesium, Taurine, Vitamins, Glucose syrup, soy protein isolate,

Sucrose, Maltodextrine, Methionine, Taurine, Carnitine, and trace elements.

Further exemplified in the Examples section below, is the preparation of

phosphatidylserine and the phospholipid complex thereof.

EXAMPLE XI

Tables 1 and 2 below provide exemplary compositions of dry and fluid milk

based infant formulas and dry and fluid soy based infant formulas according to the

present invention.

Infant Formula I (Milk-Based)

The formula comprises the following ingredients: lactose, vegetable oils, skimmed milk powder, whey protein concentrate, Sodium, Calcium, Phosphorus,

Potassium, Chloride, Iron, Magnesium, Taurine, Vitamins and phospholipids.

TABLE 1 owder fluid

Unit 100 g 100 ml

General Comp.

Protein gram 11.1 1.5

Fat gram 25.9 3.5

Lactose . gram 55.5 7.5

Water gram 2.5

Ash gram 2.06 0.27

Vitamins

Vitamin A I.U. 1500 200

Vitamin D I.U. 300 40

Vitamin E mg 6 0.81 Vitamin K . u . 15 2.01

Vitamin Bl .mu. 350 47.03

Vitamin B2 .mu. 450 60

Vitamin B6 .mu, 222 30

Vitamin B12 .mu. 0.66 0.09

Niacin mg 2 0.27

Folic Acid .mu, 45 6

Calcium mg 4.44 0.06

Pantothenate

Biotin . u. g 11 1.5

Vitamin C mg 45 6.08

Minerals

Calcium mg 326 44

Phosphorus mg 219 29

Magnesium mg 37 5

Iron mg 7.4 1

Sodium mg 120.7 16

Potassium mg 373 50

Ca/p ratio 1.49 1 49

Amino Acid Profile

Alanine mg 522 69 6

Arginine mg 368 49 1

Asparatic Acid mg 11.10 1 5

Cystine mg 191 25 5

Glutamic Acid mg 1423 189 7

Glycine mg 244 32 5

Histidine mg 262 34 9

Isoleucine mg 761 101 5

Leucine mg 12.20 1 62

Lysine mg 10.00 1 3

Methionine mg 270 36

Phenylalanine mg 461 62 3

Proline mg 962 128 3

Serine mg 681 90.8

Taurine mg 37 4.9

Threonine mg 686 91.5

Tryptophan mg 180 24

Tyrosine mg 463 61.7

Valine mg 775 103.3

Fatty Acid Profilf

Caprylic (C8) from fat 2.6 2, 6

Capric (CIO) 2.1 2, 1

Laurie (C12) 17.5 17, 5

Meristic (C14) 6.7 6, 7

Palmitic (C16) 11.2 11, 2

Stearic (C18) 11.8 11.8

Oleic (C18:l) 37.0 37.0

Linoleic (C18:2) 10.0 10.0

Linolenic (C18:3) 1.2 1.2

Phospholipid supplement which is added to the aforementioned lOOg. of powder or fluid) :

Phosphatidylcholine mg 250.0 250.0

Phosphatidylethanolamine mg 250.0 250.0

Phosphatidylserine mg 100.0 100.0

Phosphatidylinositol mg 50.0 50.0

Phosphatidic Acid mg 50.0 50.0 Infant Formula II (Soy-Based)

The formula comprises the following ingredients: Glucose syrup, vegetable

oils, soy protein isolate, Sucrose, Maltodextrine, Sodium, Calcium, Phosphorus,

Potassium, chloride, Iron, Magnesium, Vitamins, Methionine, Taurine, Camitine,

trace elements and phospholipids.

TABLE 2

Powder fluid

Unit 100 g 100 ml

General Comp .

Protein gram 15 1.98

Fat gram 27.54 3.64

Carbohydrate gram 51.5 6.8

Linoleic Acid gram 4.5 0.6

Vitamins

Vitamin A I.U. 1500 198

Vitamin D I.U. 300 39.7

Vitamin E I.U. 10 1.32

Vitamin C mg 65 8.6

Vitamin K . u. g 77 10.2

Vitamin Bl .mu.g 345 45.6

Vitamin B2 .mu.g 445 58.9

Vitamin B6 .mu. g 327 43.3

Vitamin B12 .mu.g 1.5 0.2

Niacin mg 7 0.93

Folic Acid .mu.g 76

Pantothenic Acid .mu.g 4.5 0.6

Biotin .mu.g 25 3.3

Choline mg 58 7.7

Minerals

Calcium mg 500 66.2

Phosphorus mg 300 39.7

Magnesium mg 45 6

Iron mg 9.2 1.2

Zinc mg 4 0.53

Manganese .mu.g 150 19.8

Copper .mu.g 400 53

Iodine .mu.g 77 10.2

Sodium mg 200 26.5

Potassium mg 546 72.2

Chloride mg 400 53

Inositol mg 25 3.3

Carnitine mg 10 1.3

Ca/P ratio 1.67 1.67

Amino Acid Profile

Alanine mg 640 85.3

Arginine mg 497 6.5 Aspartic Acid mg 1385 184.7

Cystine mg 242 32.3

Glutamic Acid mg 3065 408.7

Glycine mg 300 40

Histidine mg 382 50.9

Isoleucine mg 893 119.1

Leucine mg 1600 213.3

Lysine mg 1360 181.3

Methionine mg 406 54.1

Phenylalanine mg 650 86.7

Proline mg 1113 148.4

Serine mg 737 98.3

Taurine mg 51 6.8

Threonine mg 460 61.3

Tyrosine mg 621 82.8

Valine mg 947 126.3

Fatty Acid Profile

Caprylic (C8) % from fat 2.6 2.6

Capric (CIO) o. o 2.1 2.1

Laurie (C12) g, o 17.5 17.5

Meristic (C14) o 6.7 6.7

Palmitic (C16) g, o 11.2 11.2

Stearic (C18) 11.8 11.8

Oleic (C18:l) Q.

O 37.0 37.0

Linoleic (C18:2) g, o 10.0 10.0

Linolenic (C18:3) 9, o 1.2 1.2

Phospholipid supplement which is added to the aforementioned lOOg. of powder or fluid) : Phosphatidylcholine mg 250.0 250.0

Phosphatidylethanolamine mg 250.0 250.0

Phosphatidylserine mg 100.0 100.0

Phosphatidylinositol mg 50.0 50.0

Phosphatidic Acid mg 50.0 50.0

Although the invention has been described in conjunction with specific

embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all

such alternatives, modifications and variations that fall within the spirit and broad

scope of the appended claims.

EXAMPLE 2-1

Soybean lecithin (50 g; Epikuron 135 as the product name; Lucas Meyer GmbH, Germany) and soybean oil (10 g) were placed in a 300-ml vial, followed by addition of ethyl acetate (50 ml) for solubilization. Adding a solution (20 ml) of 0.30 g/ml L-serine dissolved in 0.1M sodium phosphate buffer, pH 7.0 to the resulting

solution for thorough blending, a solution of 500 U/ml phospholipase-D from cabbage was added to the mixture solution for reaction at 25. degrees C. for 5 hours under

stirring with a stirrer.

For inactivation of the enzyme in the reaction solution, the vial containing the

reaction solution was immersed in hot water. Subsequently, the reaction solution was

cooled^" in ice and treated with chloroform. The chloroform layer was colected and

dried under reduced pressure.

EXAMPLE 2-2

Using egg yolk lecithin (DS-PL95E as the product name; manufactured by .

Doosan Corp. Venture BG Biotech BU. Korea) as the substrate, rearranged phosphatidylserine was produced by the same method as in Example 1-1.

EXAMPLE 3

Phosphatidylserine was prepared by Lipogen Products (9000) Ltd. via the

process of enzymatic reaction from a substrate soybean lecithin according to Example

2-1. 50 grams of this preparation were added to 50 grams of soy lecithin containing high percentage of phosphatidyl choline (50 g; Epikuron 145 V as the product name;

Lucas Meyer GmbH, Germany). The mixture was vigorously stirred to produce a homogeneous phospholipids complex.

Claims

WHAT IS CLAIMED IS:

1. A pharmaceutical or nutritional or food composition for feeding infants, which

is more similar to human milk in a powder or fluid form, having an effect of addresseing the shortcomings of the presently known infant formulas by the inclusion

of a phospholipid supplement which contains phosphatidylserine as one of the

effective ingredients.

2. The composition of claim 1, comprising of a phospholipid complex including

phosphatidylserine or salts thereof as one of the effective ingredient, comprising at

least l% (w/w) phosphatidyserine out of the total phospholipid content of the

composition, wherein the phosphatidylserine has a structural fatty acid chain derived

from at least one raw material lecithin selected from the group consisting of soy bean lecithin, rapeseed lecithin, or egg yolk lecithin, and which is produced by reaction

with phospholipase-D.

3. The composition of claim 1, comprising of a phospholipid complex including phosphatidylserine or salts thereof as one of the effective ingredient, comprising at

least 1% (w/w) phosphatidyserine out of the total phospholipid content of the

from at least one raw material lecithin selected from the group consisting of soy bean

lecithin, rapeseed lecithin, or egg yolk lecithin and which is produced by reaction with phospholipase-D and wherein the structural fatty acid chain is a hydrogenated saturated fatty acid chain.

4. The composition of claim 1 which also contains milk or soy derived nutritional

ingredients.

5. The composition of claim 1 which may also contain one or more of the

following ingredients: lactose, vegetable oils, skimmed milk powder, whey protein

concentrate, Sodium, Calcium, Phosphorus, Potassium, Chloride, Iron, Magnesium, Taurine, Vitamins, Glucose syrup, soy protein isolate, Sucrose, Maltodextrine,

Methionine, Taurine, Carnitine, and trace elements.

6. The composition of claim 1 which also contains pharmaceutical or food

excipient.

7. The composition of claim 1 which is mixed in water for obtaining a fluid

including the nutritional components and the phospholipid supplement and which is fed to the infant.