THIS APPLICATION HAS BEEN DIVIDED OUT OF INDIAN APPLICATION
NO. 534/KOLNP/2005
INFANT FORMULA CONTAINING NUCLEOTIDES
FIELD OF THE INVENTION
The present invention is directed to infant formula compositions which contain
nucleotides. More specifically, the present invention relates to preterm or low birth
weight infant formula containing nucleotides at levels suitable for feeding preterm
infants.
DESCRIPTION OF THE PRIOR ART
Preterm infants have increased. nutrient demands in order to maintain the
rapid early life development of body tissue. In pediatric care facilities, the survival of
low birth weight infants (LBW > 1000g) and in particular, very low birth weight infants
(VLBW < 1000g) has increased. However, the endogenous nutrient storage
capabilities of these infants is severely limited.
The premature infant is notably different from the term infant. The initial
immaturity and subsequent rapid growth rate of the premature infant necessitates a
higher nutrient intake than the term infant. During the past several years,
nutritionists have made substantial progress regarding the establishment of the
nutrient needs of premature infants and have adjusted the composition of preterm
infant formulas accordingly. The composition of preterm infant formula is based
primarily on the composition of human breast milk as well as on intra-uterine
accretion. Preterm infant formulas have thus been modified to contain appropriately
increased amounts of nutrients, such as protein and fat, as well as minerals, such as
calcium, phosphorus, copper and zinc.
Means for providing adequate nutrition to premature infants has long been
desired. Efforts have been made to enrich standard infant formula in such a manner
to enrich its nutrition capabilities particularly in the feeding of premature infants.
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Nucleotides are found in all living cells. Mature human breast milk contains
thirteen different nucleotides. The five nucleotides which appear in the highest
concentration are cytidine monophosphate (CMP), guanosine monophosphate
(GMP), adenosine monophosphate (AMP), undine monophosphate (UMP), and
inosine monophosphate (IMP). Human milk further comprises nucleosides,
nucleotide polymers and nucleotide adducts capable of contributing to the Total
Nucleotide Pool (TNP). However, in the preterm infant, de novo synthesis and the
salvage method for nucleotide formation may be inadequate.
Infant formulas for term infants have been fortified with nucleotides to
simulate mother's milk and to treat certain specific conditions.
For example, US Patent No. 5,492,899 discloses an infant formula containing
CMP, UMP, AMP and GMP to provide a dietary formula that enhances the immune
system and alleviates diarrhea.
US Patent No. 4,544,559 discloses a nucleotide enriched humanized milk for
infant nourishment which contains AMP, CMP, GMP, IMP and UMP at specific
levels.
US Patent Nbs. 4,994,442 and 5,086,500 disclose the addition of nucleosides
and nucleotides to infant formulas having enhanced physiological properties and
which closely resemble mother's milk.
However, the addition of nucleotides to infant formula to resemble the milk of
mother's of preterm infants has not been disclosed. It is postulated that nucleotides
obtained from the diet may be essential to the preterm infant playing an important
role in cell growth, energy transfer, and promotion of gut maturation and
development. Nucleotides may also provide benefits relating to biological effects,
including increasing beneficial gut microflora, improving lipid metabolism and
enhancing immunity.
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SUMMARY OF THE INVENTION
The present invention is directed to an infant formula composition comprising
a total nucleotide concentration similar to that of the breast milk of mothers of
preterm infants. The present infant formula compositions comprise 3.2 mg/L to 15.4
mg/L of CMP; 1.8 mg/L to 11.0 mg/L of UMP; 1.8 mg/L to 8.0 mg/L of GMP; 0.1
mg/L to 2.2 mg/L of IMP; and 2.5 mg/L to 13.2 mg/L of AMP.
DETAILED DESCRIPTION OF THE INVENTION
The present infant formula compositions have a total nucleotide
concentration of 9.3 mg/L to 49.8 mg/L, preferably 12.4 mg/L to 37.4 mg/L and most
preferably 15.5 mg/L to 24.9 mg/L.
The present infant formula compositions comprise the following nucleotides
in the stated amounts:
a) CMP: 3.2 mg/L to 15.4 mg/L, preferably 4.2 mg/L to 11.6 mg/L; and
most preferably 5.3 mg/L to 7.7 mg/L;
b) UMP: 1.8 mg/L to 11.0 mg/L, preferably 2.4 mg/L to 8.3 mg/L, and
most preferably 3.0 mg/L to 5.5 mg/L;
c) GMP: 1.8 mg/L to 8.0 mg/L, preferably 2.4 mg/L to 6.0 mg/L, and most
preferably 3.0 mg/L to 4.0 mg/L;
d) IMP: 0.1 mg/L to 2.2 mg/L, preferably 0.1 mg/L to 1.7 mg/L, and most
preferably 0.1 mg/L to 1.1 mg/L; and
e) AMP: 2.5 mg/L to 13.2 mg/L, preferably 3.3 mg/L to 9.9 mg/L, and
most preferably 4.1 mg/L to 6.6 mg/L.
The present infant formulas may be additionally formulated with other
nutritionally beneficial ingredients recognized in the art, such as polyunsaturated fatty
acids, (e.g. arachidonic acid and docosahexaenoic acid). The present formulas may
also contain all vitamins and minerals essential in the diet of the preterm infant.
These vitamins and minerals should be present in nutritionally significant amounts.
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Examples of vitamins and minerals which may be added to the present infant
formula compositions include vitamin A, vitamin B complex, vitamin C, vitamin D,
vitamin E, vitamin K, calcium, magnesium, sodium, potassium, phosphorous, copper,
zinc, chloride, iodine, selenium, iron, niacin, folic acid, pantothenic acid, biotin,
choline, inositol and manganese.
The infant formula compositions of the present invention may further
comprise macronutrients beneficial to the preterm infant such as carotenoids,
taurine, coconut oil, sunflower oil, soy oil, native or synthetic fats, palmitic acid,
medium chain triglycerides, lactose, maltodextrin, glucose, bovine oligosaccharides
or fractions thereof, skim milk, milk whey, soy protein, rice protein, protein
hydrolysates and lactoferrin.
The present infant formulas, which are particularly suited for feeding the
preterm or low birth weight infant, contain nucleotides at levels found in breast milk
of mothers of preterm infants. These levels are quite different than the levels of
nucleotides found in the breast milk of mothers of term infants.
The present infant formulas may be in the form of a liquid, either as a ready
to feed liquid or as a concentrated liquid requiring dilution with water prior to feeding,
or in a powder form requiring reconstitution with water prior to use.
The present infant formula may be prepared by blending appropriate
quantities of whey protein concentrate with skimmed milk, lactose, vegetable oils and
fat soluble vitamins in deionized water. Preferably, these materials are blended
together in quantities sufficient to provide a final concentration of approximately 240
grams/liter. Mineral salts may then be added to the mixture prior to a high
temperature/short time pasteurization step. Appropriate mineral salts include
calcium chloride, calcium phosphate, sodium citrate, potassium hydroxide,
potassium bicarbonate, ferrous sulfate, zinc sulfate, sodium chloride, copper sulfate,
potassium iodide, sodium selenite, eta. The mixture is then homogenized and
cooled. Heat-labile vitamins and micronutrients may then be added to the mixture.
The mixture is then standardized with deionized water to a final total solids
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concentration of about 150 to about 160 and preferably about 155 grams per litre,
which is equivalent to about 820 kcal per liter. The formula may be sterilized using a
conventional ultrahigh temperature process, and then aseptically filled into
appropriate packaging. Alternatively, the formula may be filled into glass jars and
then retort sterilized.
It would be recognized by one skilled in the art that other known methods of
manufacture and sterilization can be used for the preparation of the present infant
formula. The present infant formula may also be produced as a concentrated liquid
product requiring dilution with an equal volume of water prior to feeding to an infant.
Furthermore such an infant formula may be dehydrated, such as in a spray dryer, to
create a stable infant formula powder that offers advantages of stability and
economy of transport, said powder requiring reconstitution with water prior to feeding
to an infant.
EXAMPLES
Example 1
A study was conducted to determine the total nucleotide concentration and
the concentration of individual nucleotides in the breast milk collected from mothers
of newborn, premature infants. The study was a prospective, non-randomized open
study. Twenty seven mothers provided milk samples of volume greater than or
equal to 20 ml.
The inclusion criteria for the study were as follows: outpatient, healthy
women who were 18 to 40 years of age; parity less than 5; lactating; and between
4-18 days post partum on the scheduled day of collection. The study included
mothers whose infants were born between 26-32 weeks gestational age, who were
appropriate for gestational age and who weighed less than 1800 grams at birth.
Samples of preterm human milk were analyzed for free nucloeosides and
free nucleosides after incubation with alkaline phosphatase. The increase in free
nucleosides after the enzyme treatment represents the monomeric nucleotide
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content on a molar basis. The concentrations in Table 1 below are expressed as
equivalent monophosphates.
The results of this analysis are set forth in Table 1:
Table 1

Range (mg/L) Mean (mg/L)
CMP 0.46-14.48 5.3
UMP 0.29 - 6.65 3.0
GMP 0.87-10.16 4.8
IMP 0.00-1.35 0.1
AMP 1.82-8.55 4.1
Total monomeric nucleotide 3.44-41.19 17.3
Example 2
An example of the present infant formula is set forth below.
A typical 4,000 L production batch of ready-to-feed liquid infant formula
mixture was manufactured in the following manner.
A fat blend was prepared by combining 86.5 kg of Betapol (Loders Croklaan)
36.1 L of a high-oleic sunflower oil or safflower oil, 8.94 L of soybean oil, 24.7 kg of
medium chain triglycerides, 1.36 kg of soy lecithin, 2.47 kg of an oil containing 38%
of its fatty acids as arachidonic acid (ARASCO, Martek, Columbia, MD) and 1.65 kg
of an oil containing 38% of its fatty acids as docosahexaenoic acid (DHASCO,
Martek, Columbia, MD).
The fat-soluble vitamins, vitamin A (vitamin A palmitate), beta-carotene,
vitamin D (cholecalciferol), vitamin E (dl-alpha-tocopherol acetate) and vitamin K
(phytonadione) were then dispersed in the fat blend.
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An amount of heated liquid skim milk or powdered skim milk sufficient to
provide 44 kg of protein was added to warm deionized water in a compounding tank
to which had been added 3620 g of sodium citrate and 1590 g potassium
bicarbonate dissolved in hot water. The fat blend was metered into the compounding
tank. The major dry ingredients, lactose, whey protein concentrate and maltodextrin,
were added through a powder funnel eductor. The amount of added whey protein
concentrate varied according to its protein content and was sufficient to provide 44
kg of protein. Taurine, 221 g was dissolved in hot water and added to the
compounding tank. The following minerals were dissolved or dispersed separately in
hot water and added to the compounding tank with intensive agitation: calcium
phosphate, 2220 g; calcium chloride, 1710g; sodium chloride, 512g; potassium
hydroxide, 643 g; ferrous sulfate, 153 g; potassium iodide, 72.5 g of a 1% triturate in
lactose; 359 g of a 44.6% solution of zinc sulfate; 598 g of a 2.5% solution of copper
sulfate and 7.22 g of a 0.3% sodium selenite triturate in potassium bicarbonate.
The mixture was heated to 96 + 2°C, held for 30 seconds and then cooled to
66 ± 2°C. The mixture was then homogenized in a two-stage homogenizer, at 2500
psig in the first stage and 500 psig in the second stage. The homogenized mixture
was then cooled to 5 - 10°C using a plate heat exchanger.
A sample of the mixture was analyzed for fat, pH and total solids. An
appropriate amount of deionized water was added as required to achieve 80% of
total dilution and the batch was mixed. A sample was taken for analysis of total solids
and pH. The quantity of water required for final dilution was calculated. A portion of
this dilution water was used to dilute the nucleotides and water-soluble vitamins
before their addition to the mix. The following nucleotides were dissolved in warm
water and added to the mix: cytidine 51 monophosphate, 31.7 g; adenosine 5'
monophosphate, 27.9 g; uridine 5' monophosphate, disodium salt, 29.7 g; inosine 5'
monophosphate, disodium salt, 5.43 g; and guanosine 5' monophosphate, disodium
salt, 22.4 g.
Water-soluble vitamins were dissolved in warm water and were added to the
mix as follows: Nicotinamide, 109 g; calcium pantothenate, 42.3 g; Thiamine
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hydrochloride, 8.98 g; Riboflavin, 3.55 g; Pyrodoxine hydrochloride, 5.8 g; Folic acid,
2.51 g; D Biotin, 0.113 g; Cyanocobolamin, 7.24 g and myo-inositol, 104 g, were
dissolved in water and added to the mix. Ascorbic acid, 1180 g, was dissolved in
warm water and 535 g of potassium bicarbonate were added slowly to neutralize the
ascorbic acid. The neutral ascorbate solution was then added to the mix. Citric acid,
106 g was dissolved in warm water and added to the mix. The mix was agitated for
30 minutes. The mix was standardized to total solids of about 123 g per litre.
Twenty hours after the initiai homogenization, the mix was homogenized
again and sterilized through a Stork UHT (ultra high temperature) aseptic processing
system and filled into 100 ml glass jars.
The glass jars were then retort sterilized in a Barriquand rotary steriliser.
The ingredients of this formula were as follows:

Inqredients Units /100 ml /100 kcal
Protein g 2.0 2.44
Fat g 4.4 5.37
Carbohydrate g 8.6 • 10.5
Linoleic Acid mg 550 671
Energy kcal 82 100
Vitamin A meg '90 110
Vitamin D3 meg 1.5 1.8
Vitamin E mg 1.2 1.5
Vitamin K1 meg 8 9.76
Vitamin B1 meg 120 146
Vitamin B2 meg 200 244
Vitamin B6 meg 72 88
Vitamin B12 meg 0.3 0.37
Niacin (NE) meg 820 1000
Folic Acid meg 48 59
Pantothenic Acid meg 450 549
Biotin meg 2.4 2.9
Vitamin C mg 11 ' 13.4
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Calcium mg 80 97.6
Phosphorus mg 42.5 51.8
Magnesium mg 8 9.76
Iron mg 0.8 0.98
Zinc mg 0.8 0.98
Manganese meg 10 12.20
Copper meg 82.5 101
Iodide meg 10 12.2
Sodium mg 35 42.7
Potassium mg 85 104
Chloride mg 60 73.2
Choline mg 15 18.3
Inositol mg 4.5 5.49
Taurine mg 4.8 5.9
CMP mg 0.7 0.9
UMP mg 0.5 0.6
GMP mg 0.4 0.5
IMP mg 0.1 0.12
AMP mg 0.6 0.7
Example 3
The following infant formula acccording ^,to the present invention was
prepared according to the process described in Example 2.

Inqredients Units /100 ml MOO kcal
Protein g 2.20 2.70
Fat g 4.23 5.20
Carbohydrate g 8.62 10.6
Linoleic Acid mg 550 677
Energy kcal 81.3 100
Vitamin A meg 224 275
Vitamin D3 meg 3.48 4.28
Vitamin E mg 4.09 5.03
Vitamin K1 meg 6.1 7.5
Vitamin B1 meg 142 175
Vitamin B2 meg 203 250
Vitamin B6 meg 122 150
Vitamin B12 meg 0.33 0.41
Niacin (NE) meg 2353 2870
Folic Acid meg 40.6 50.0
Pantothenic Acid meg 1016 1250
Biotin meg 3.25 4.00
Vitamin C mg 14.6 18.0
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Calcium mg 102. 125
Phosphorus mg 61 75.0
Magnesium mg 8.1 10.0
Iron mg 0.81 1.00
Zinc mg 0.81 1.00
Manganese meg 4.1 5.0
Copper meg 91 112
Iodide meg 24 30.0
Sodium mg 40.6 50.0
Potassium mg 81 100
Chloride mg 73.2 90.0
Selenium meg 1.71 2.10
Choline mg 13.0 16.0
Inositol mg 30.08 37.00
Taurine mg 4.80 5.90
CMP mg 0.7 0.9
UMP mg 0.5 0.6
GMP mg 0.4 0.5
IMP mg 0.1 0.12
AMP mg 0.6 0.7
The present invention may be embodied in other specific forms without
departing from the spirit and essential attributes thereof and accordingly, reference
should be made to the appended claims, rather than to the foregoing specification,
as indicating the scope of the invention.
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WE CLAIM;
1. An infant formula composition comprising 3.2 mg/L to 15.4 mg/L of
CMP; 1.8 mg/L to 11.0 mg/L of UMP; 1.8 mg/L to 8.0 mg/L of GMP; 0.1 mg/L to 2.2
mg/L of IMP; and 2.5 mg/L to 13.2 mg/L of AMP.
2. The infant formula composition of claim 1, comprising 4.2 mg/L to
11.6 mg/L of CMP; 2.4 mg/L to 8.3 mg/L of UMP; 2.4 mg/L to 6.0 mg/L of GMP; 0.1
mg/L to 1.7 mg/L of IMP; and 3.3 mg/L to 9.9 mg/L of AMP.
3. The infant formula composition of claim 2, comprising 5.3 mg/L to 7.7
mg/L of CMP; 3.0 mg/L to 5.5 mg/L of UMP; 3.0 mg/L to 4.0 mg/L of GMP; 0.1 mg/L
to 1.1 mg/L of IMP; and 4.1 mg/L to 6.6 mg/L of AMP.
4. An infant formula composition having a total nudeotide concentration
of 9.3 mg/L to 49.8 mg/L.
5. The infant formula composition of claim 4 having a total nudeotide
concentration of 12.4 mg/L to 37.4 mg/L.

6. The infant formula composition of claim 5 having a total nudeotide
concentration of 15.5 mg/L to 24.9 mg/L.
7. A method of feeding preterm infants comprising feeding said infant a
nutritionally sufficient amount of the infant formula composition of claim 1.
8. An infant formula composition substantially is herein described particularly
with reference to the examples.


Dated this 4th day of June 2007.
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Infant formula compositions are provided which comprise 3.2mg/L to 15.4mg/L of CMP; 1.8 mg/L to 11.0mg/L of UMP; 1.8 mg/L to 8.0 mg/L of GMP; 0. 1 mg/L to 2.2 mg/L of IMP; and 2.5 mg/L to 13.2 mg/L of AMP.