U.S. patent application number 11/912925 was filed with the patent office on 2009-01-08 for polar lipid mixtures, their preparation and uses.
Invention is credited to Gai Ben Dror, Yael Herzog, Dori Pelled, Avidor Shulman, Yoni Twito, Rassan Zuabi.
Application Number | 20090011075 11/912925 |
Document ID | / |
Family ID | 36645701 |
Filed Date | 2009-01-08 |
United States Patent
Application |
20090011075 |
Kind Code |
A1 |
Shulman; Avidor ; et
al. |
January 8, 2009 |
Polar Lipid Mixtures, their Preparation and Uses
Abstract
Disclosed herein are polar lipid mixtures, comprising
glycerophospholipids such as phosphatidylcholine (PC),
phosphatidylethanolamine (PE), phosphatidylserine (PS) and
phosphatidyl-inositol (PI), and sphingolipids such as sphyngomyelin
(SM). Most importantly, the ratio of phospholipids in said mixture
is comparable to that of HMF, and is represented by
SM>PC>PE>PS>PI or SM=PC>PE>PS>PI. Processes
for the preparation of said mixtures and uses thereof are also
described herein.
Inventors: |
Shulman; Avidor; (Kiryat
Tivon, IL) ; Zuabi; Rassan; (Kfar Neen, IL) ;
Ben Dror; Gai; (Moshav Ofer, IL) ; Twito; Yoni;
(Geva Carmel, IL) ; Pelled; Dori; (Hod Hasharon,
IL) ; Herzog; Yael; (Nesher, IL) |
Correspondence
Address: |
Fleit Gibbons Gutman Bongini & Bianco PL
21355 EAST DIXIE HIGHWAY, SUITE 115
MIAMI
FL
33180
US
|
Family ID: |
36645701 |
Appl. No.: |
11/912925 |
Filed: |
April 27, 2006 |
PCT Filed: |
April 27, 2006 |
PCT NO: |
PCT/IL06/00510 |
371 Date: |
May 12, 2008 |
Current U.S.
Class: |
426/2 ; 426/580;
426/608; 426/648 |
Current CPC
Class: |
A23J 7/00 20130101; A61P
25/28 20180101; A23V 2250/1852 20130101; A23V 2250/185 20130101;
A23V 2250/1848 20130101; A23V 2002/00 20130101; A23V 2002/00
20130101; A61P 25/00 20180101; A23L 33/115 20160801 |
Class at
Publication: |
426/2 ; 426/608;
426/580; 426/648 |
International
Class: |
A23L 1/30 20060101
A23L001/30 |
Foreign Application Data
Date |
Code |
Application Number |
Apr 28, 2005 |
IL |
168294 |
Claims
1-56. (canceled)
57. A man-made lipid preparation comprising a mixture of
glycerophospholipids being phosphatidylcholine (PC),
phosphatidylethanolamine (PE), phosphatidylserine (PS) and
phosphatidyl-inositol (PI), and optionally comprising a
sphingolipid or a precursor or metabolite thereof, wherein the
level of each of said PC, PE, PS and PI is at least 1% w/w, the
ratio between said glycerophospholipids is PC>PE>PS>PI,
and said glycerophospholipids are derived from a natural non-brain
animal lipid source.
58. The lipid preparation of claim 57, wherein said sphingolipid is
sphingomyelin (SM).
59. The lipid preparation of claim 58, comprising sphingomyelin,
wherein the ratio between said glycerophospholipids is
SM>PC>PE>PS>PI.
60. The lipid preparation of claim 57, wherein said
glycerophospholipids are obtained from said natural non-brain
animal lipid source by fractionation and/or extraction.
61. The lipid preparation of claim 57, wherein the level of PC is
at least 5% w/w, the level of PS is at least 2% w/w, the level of
PE is at least 4% w/w, and the level of PI is at least 2% w/w.
62. The lipid preparation of claim 59 comprising at least 2% w/w
SM.
63. A process for the preparation of a lipid preparation comprising
a mixture of glycerophospholipids being phosphatidyl-choline (PC),
phosphatidylethanolamine (PE), phosphatidylserine (PS) and
phosphatidyl-inositol (PI), and optionally comprising a
sphingolipid or a precursor or metabolite thereof preferably
sphingomyelin, wherein the ratio between said glycerophospholipids
is PC>PE>PS>PI, from a natural non-brain animal lipid
source, comprising isolating said lipid mixture from said natural
non-brain animal lipid source by methods which do not involve
transphosphatidylation.
64. The process of claim 63, wherein said sphingolipid is
sphyngomyelin.
65. The process of claim 64, wherein said glycerophospholipid
mixture comprises sphingomyelin from a natural non-brain animal
lipid source, wherein the ratio between said glycerophospholipids
is SM>PC>PE>PS>PI.
66. The process of claim 63, wherein said isolation comprises the
steps of: (a) providing a natural non-brain animal lipid source
which has a substantially low content of polar lipids; (b) removing
non-lipid material from said lipid source, dispersing the lipids,
preferably with agitation, in a suitable organic solvent or a
mixture of organic solvents; (c) separating the dissolved lipid
fraction obtained in step (b) and removing the organic solvent
therefrom to give a lipid fraction; (d) de-oiling said lipid
fraction obtained in step (c) at least once to remove any non-polar
lipids; (e) filtering and drying said polar lipids obtained in step
(d); and further optionally comprising a step of treating said
lipid source with an aqueous medium, either before or after step
(b).
67. The process of claim 66, wherein said glycerophospholipids
mixture comprises sphingomyelin from a natural non-brain animal
lipid source, wherein the ratio between said glycerophospholipids
in said preparation is SM>PC>PE>PS>PI, comprising the
steps of: (a) providing a natural non-brain animal lipid source
which has a substantially low content of polar lipids; (b) removing
non-lipid material from said lipid source, dispersing the lipids,
preferably with agitation, in a suitable organic solvent or a
mixture of organic solvents, (c) separating the dissolved lipid
fraction obtained in step (b) and removing the organic solvent
therefrom to give a lipid fraction; (d) de-oiling said lipid
fraction obtained in step (c) at least once to remove any non-polar
lipids; (e) filtering and drying said polar lipids obtained in step
(d); and further optionally comprising a step of treating said
lipid source with an aqueous medium, either before or after step
(b).
68. The process of claim 66, wherein said natural lipid source is
derived from bovine milk, and contains up to 5% w/w, preferably 10%
w/w, more preferably 25% w/w, even more preferably 35% w/w of total
lipids, and optionally contains sphingomyelin, in addition to other
constituents including proteins and carbohydrates, and wherein
about 20%, preferably 30%, more preferably 50%, most preferably 70%
of said total lipids are polar lipids.
69. The process of claim 64, said process comprising further
subjecting said preparation to a step of any one of alkaline
hydrolysis, enzymatic hydrolysis, preparative chromatography or
polar lipid extraction, whereby the lipid preparation obtained is
enriched with sphingomyelin (SM).
70. A dietary supplement comprising a lipid preparation as defined
in claim 57.
71. The dietary supplement of claim 70, for use as an ingredient of
a lipid constituent of infant formulas or as an ingredient of
infant formulas.
72. A method for the enhancement of infants and/or children
development, comprising administering to said infant or child a
lipid preparation as defined in claim 57.
73. The method of claim 72, wherein said development is cognitive
development or vision development.
74. A food article comprising the dietary supplement of claim
70.
75. The food article of claim 74, wherein said food article is an
infant formula.
76. The process of claim 63, further comprising the step of
isolating SM from the polar lipid fraction.
77. A method for the treatment of brain related illnesses or
disorders or for improving cognitive functions or for treating,
preventing or ameliorating myelin-related disorders or diseases,
particularly de-myelination related disorders in a subject in need
of such treatment, comprising administering to said subject the
lipid preparation as defined in claim 57.
78. The method of claim 77, wherein said brain-related disorder is
one of mood-, memory-, stress-, or age-related disorders, dementia,
Alzheimer's disease, said cognitive function is one of memory loss,
problems of concentration and attention and learning capabilities,
and said de-myelination related disorder is MS.
79. A method of supporting and enhancing in a subject, the normal
or improved development of myelin sheath and other
sphingomyelin-related tissues comprising administering to said
subject the lipid preparation of claim 58.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to polar lipids preparations,
in particular mixtures comprising glycerophospholipids, optionally
with sphingomyelin, their preparation and uses.
BACKGROUND OF THE INVENTION
[0002] All publications mentioned throughout this application are
fully incorporated herein by reference, including all references
cited therein.
[0003] Human milk fat (HMF) is composed of about 30-40 g/L lipids.
Of those, approximately 98% are triglycerides, 0.3-1%
phospholipids, and 0.4% cholesterol. The phospholipids are composed
of four major moieties: sphingomyelin (SM), phosphatidylcholine
(PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), and
phosphatidylinositol (PI).
[0004] Although glycerophospholipids, sphingomyelin, cholesterol
and their derivatives are found in relatively small amounts in
mother's milk, they play an important role in the nutrition of
developing infants, and have essential roles in all physiological
systems and cycles of the human body.
[0005] Total fat content increases gradually from colostrum (2.0%)
through transitional (2.5% to 3.0%) to mature milk (3.5% to 4.5%)
[Bitman et al. (1983) Am. J. Clin. Nutr. 38:300-312].
[0006] The role of phospholipids, and especially the role of the
phospholipid backbone, in human breast milk is poorly understood.
Most scientific research on human breast milk phospholipids in fact
uses them as an assay for the intake and incorporation of different
fatty acids on the phospholipid skeleton.
[0007] Phospholipids are involved in the structure of human milk
fat globules membrane (HMFGM), representing 23% of the membrane
mass. Interestingly, in contrast to the significant changes in
fatty acid composition from woman to woman, related mainly to race
and diet, the phospholipid composition remains constant, and is not
influenced by diet. Essentially, the level of phospholipids in
human breast milk only changes with the age of the infant. This
further suggests that phospholipids are an essential nutritional
component of human breast milk.
[0008] Phospholipids show a decrease from high levels in colostrum
(1.1% out of the total fat fraction) to lower levels in mature milk
(0.6%). The decline in phospholipids is consistent with an increase
in the fat globule size [Ruegg et al. (1981) Biochim. Biophys. Acta
666, 7-14]. The phospholipids composition of breast milk from
mothers of term and preterm infants during lactation was thoroughly
studied by Bitman et al. [Bitman et al. (1984) Am. J. Clin. Nutr.
40, 1103-1119].
[0009] Milk phospholipids do not exhibit any marked differences
attributable to length of gestation after day 21. This remarkable
constancy in class distribution of phospholipids indicates that the
composition of the membrane of the milk fat globules is identical
at all stages of lactation.
[0010] The amount of phospholipids (sphingomyelin and
glycerophospholipids) in human milk fat is about 15-20 mg/dL.
Sphingomyelin (SM) is found at about 37% of total polar lipids,
phosphatidylcholine (PC) is found at 28% of total polar lipids,
phosphatidyl-ethanolamine (PE) is found to be about 19%,
phosphatidylserine (PS) at 9% and phosphatidylinositol (PI) at 6%.
Thus, the ratio between the polar lipids of HMF is as follows:
SM>PC>PE>PS>PI.
[0011] Some glycerophospholipids, and especially those extracted
from soybean, are used as dietary supplements and a variety of
health benefits are associated with their intake, including
improvement of cognitive functions, as well as of memory and
concentration, maintenance of cellular membrane composition, and
contribution to general well-being. Phospholipids and lecithins are
a source of choline and they enhance the bio-availability of other
nutrients and therapeutic agents.
[0012] In addition, glycerophospholipids are used as food
emulsifiers, anti-oxidants, stabilizers, as well as in other food
application such as mold-release and anti-caking agents. They
confer unique physical properties to food products as well as
personal care products, and thus are also used in pharmaceutical
formulations as carriers and delivery systems.
[0013] WO 03/105609 describes a phospholipid supplement which
contains PS at a concentration of at least 1% out of the total
phospholipid content of the composition. Moreover, said PS is
derived from soybean lecithin, rapeseed lecithin or egg yolk, and
is enzymatically produced using phospholipase-D.
[0014] U.S. Pat. No. 5,709,888 presents fat mixtures comprising
phospholipids and LC-PUFA, such as oleic acid, linoleic acid and
alpha-linolenic acid, having an adequate level of LC-PUFA of both
the n6 and n3 series.
[0015] EP 484,266 describes a mixture of phospholipids obtained
from domestic animal brain sources, in addition to at least one of
vegetable oil, animal fat, fish oil, and/or medium chain
triglycerides, in which the ratios between LC-PUFA and
phospholipids is similar to those of human milk and Mediterranean
diet.
[0016] Most of the sphingomyelin in milk fat is a building block of
the milk fat globule membrane. Sphingomyelin is also an important
building block required by the infant for the development of the
brain and other tissues, as well as being important in several
biochemical pathways.
[0017] Myelin is the white matter coating nerve cells, enabling
them to conduct impulses between the brain and other parts of the
body. It consists of a layer of proteins, packed between two layers
of lipids. Myelin is produced by specialized cells:
oligodendrocytes in the central nervous system, and Schwann cells
in the peripheral nervous system. Myelin sheaths wrap themselves
around axons, the threadlike extensions of neurons that make up
nerve fibers. Each oligodendrocyte can myelinate several axons.
Myelin is comprised of 80 percent lipids and 20 percent proteins. A
major lipid of this important tissue is sphingomyelin. This fatty
substance protects the fiber-like axons and speeds electrical
signals as they travel along nerve pathways to carry out vital
functions such as movement.
[0018] Different diseases and syndromes are related to disorders in
myelin damage. Multiple sclerosis, for example, causes myelin to
disintegrate, causing an obstruction of signal flow, which
progressively leads to the loss of motor coordination and other
functions.
[0019] The development of myelin sheaths and neuronal network in
infants is thus crucial and depends on the availability of
appropriate lipid building blocks, either from self biosynthesis or
from dietary sources, i.e. human breast milk.
[0020] Sphingomyelin (SM) is composed of phosphocholine as the
polar head group and sphingosine as the backbone of the molecule,
and it is therefore classified as a sphingolipid. These molecules
are involved in the regulation of cell growth, cell
differentiation, and various other functions, including
cell-substratum interactions and intracellular signal transduction.
Although many foods contain a small amount of SM, its nutritional
and physiologic roles have not been fully examined.
[0021] The myelin of the Central Nervous System (CNS myelin) has a
higher lipid content (65-80%) than that of general cell membranes.
SM and sphingolipid metabolites, such as cerebrosides and
sulfatides, are prominent components of the myelin sheath. This
sheath acts as an insulator for nerve impulses and controls the
salutatory mode of conduction via the nodes of Ranvier. Myelination
of the human CNS begins from 12 to 14 wk of gestation in the spinal
cord and continues into the third decade of life in the
intracortical fibers of the cerebral cortex, but the most rapid and
dramatic changes occur between midgestation and the end of the
second postnatal year [Brody B A et al. (1987) J. Neuropathol. Exp.
Neurol. 46: 283-301; Kinney H C et al. (1988) J. Neuropathol. Exp.
Neurol. 47: 217-234]. Myelination accounts for a large part of the
more than tripling of brain weight that occurs during this
period.
[0022] A metabolic pathway for sphingolipids has been reported
[Luberto C. and Hannun Y A (1999) Lipids 34 (suppl): S5-S11]. SPT
(EC 2.3.1.50) is the first step and the rate-limiting enzyme in
sphingolipid biosynthesis, catalyzing the synthesis of
3-ketosphinganine from L-serine and palmitoyl-CoA. This enzyme is
located in the endoplasmic reticulum or Golgi apparatus. A recent
study showed that SPT activity gradually increases from the third
prenatal to the third postnatal week in the hypothalamus of rats.
As myelination begins at the same period in these animals, it is
conceivable that an increment of SPT activity may be one of the
major factors involved in myelinogenesis.
[0023] CNS myelin has a high cerebroside content when compared to
its level in other tissues. Cerebroside is generated from ceramide
by ceramide UDP-galactosyltransferase, which is the key enzyme in
the biosynthesis of cerebrosides and catalyzes the transfer of
galactose from UDP-galactose to ceramide. In rats, cerebroside is
hardly detectable in the brain before 10 d after birth, but the
cerebroside content increases markedly from the second to the third
postnatal weeks, especially between day 14 and 23 of life. Because
the period of maximum cerebroside biosynthesis corresponds with the
time of most active myelination, cerebroside is generally
recognized as a universal marker of CNS myelination.
[0024] Ceramides can be generated from L-serine and palmitoyl-CoA
through de novo synthesis by the enzyme SPT, and from SM by
sphingomyelinase. Therefore, it was hypothesized by Oshida et al.
(Pediatric Research 2003, 53:589-593) that during the period of low
SPT activity, cerebroside in CNS myelin of developing rats may be
mainly derived from dietary SM ingested in milk that is transformed
to ceramide and then to cerebroside. Oshida et al. have shown that
when the activity of SPT was inhibited through the administration
of an appropriate inhibitor, causing a decrease of cerebrosides in
rat CNS myelin, the normal maturation and weight of the myelin
tissue could be maintained through the administration of dietary
SM. Dietary supplementation of SM restored the brain weight and
myelin dry weight that were decreased by the SPT inhibitory
treatment. Furthermore, electron microscopy showed that the axon
diameter of the inhibited group was restored following the
introduction of dietary SM. These findings suggest that orally
ingested SM is transformed to ceramide or other metabolites in the
intestinal tract, which are absorbed from the bowel and entered the
circulation to reach the CNS across the blood-brain barrier.
[0025] Although sphingomyelin has been thought to be an inert
constituent of cell membranes, current studies suggest that
metabolites of sphingomyelin are involved in signal transduction
pathways. SM plays a key role in the regulation of cellular
processes. Dietary SM can contribute to myelination of developing
CNS and protect against toxic and inhibitory conditions. SM is also
a building block to other lipids, such as ceramides.
[0026] Breast-feeding seems to contribute to rapid growth of brain
weight, which is mostly the result of myelination. At birth there
is very little myelin, but by 3 years, most axons have myelin
coatings.
[0027] Since the human milk fat globule membrane resembles the
structure and/or function of cell membranes, it is curious that its
level of SM is so high, suggesting that the presence of SM can be
attributed not just to its role of membrane building block, but
also as a dietary source for SM. This role is of great importance
at early stages of gestation in order to provide SM for myelin
build-up prior to the completion of the biosynthetic route of
cerebrosides, as described above. Indeed, the higher levels of
phospholipids in general, and of SM in particular, at the early
stages of gestation may not be coincidental but rather required to
supply dietary SM to compensate the just developing biosynthetic
machinery of cerebrosides.
[0028] Sphingomyelin is usually not produced on commercial levels,
and it is produced only from animal sources, such as bovine milk,
eggs, or animal brain. Animal sources, especially those related to
brain tissues, are of course avoided in infant nutrients due to the
risk of prion disease. In most cases, sphingomyelin of animal
sources is produced at high levels of purity, mainly for purposes
of analytical standards and for scientific research. These high
purity sphingomyelin preparations are characterized by their
extremely high cost and scarce availability, and thus are also not
feasible for general popular consumption.
[0029] Recently, the dairy industry has started to utilize dairy
waste to produce nutritional preparations which contain milk
proteins, carbohydrates and small amounts of lipids. The latter
include neutral lipids as well as polar lipids, including
glycerophospholipids as well as sphingolipids, among them
sphingomyelin. These preparations contain extremely low levels of
sphingomyelin and phosphatidylserine, making them incompatible as
an industrial source for these nutrients. A typical preparation
(e.g. SM3 Powder, produced by S. A. Corman of Belgium), contains
about 4% w/w of PC, 3.2% w/w of PE, 1.6% w/w of PS, 0.9% w/w of PI
and about 2.6% w/w of SM. Such low levels would entail use of very
large quantities of such preparations in order to deliver even
small amount of PS and sphingomyelin. Furthermore, such large
quantities would result in the delivery of non-required and
non-desired proteins and carbohydrates, the latter mainly in the
form of lactose.
[0030] The above described commercial milk lipids preparations,
although having some similarity to HMF polar lipids, still differ
from the latter. The ratio between the polar lipids in the above
commercial milk-derived preparations is PC>PE>SM>PS>PI,
while in HMF, the ratio between the polar lipids is
SM>PC>PE>PS>PI. Particularly, in HMF the level of
sphingomyelin is always higher than that of PC, the ratio being of
about 1.3, while in the above commercial milk-derived preparations,
this ratio is about 0.65.
[0031] Rombaut and colleagues provide phospholipid compositions of
several dairy products [Rombaut et al. (2005) J. Dairy Sci.
482:488]. None of the tested dairy products gave a polar lipids
ratio that is corresponding to the SM>PC>PE>PS>PI ratio
of found in HMF.
[0032] As can be seen in Table 1, the lipid composition of a
typical commercial milk-derived preparation differs from HMF mainly
in the level of sphingomyelin, which is lower than the level of PC,
and in the higher level of PE.
TABLE-US-00001 TABLE 1 Commercial milk lipids HMF lipids Lipid
class (% from total polar lipids) (% from total polar lipids) SM
21.1 37.5 PC 32.5 28 PE 26.0 19.5 PS 13.0 9 PI 7.4 6 Total 100
100
[0033] In WO 2005/051091, the present inventors described a
composition that mimics the phospholipid composition of human
breast milk. The present invention concerns the polar lipids of
human breast milk and the importance of their supplementation by
other sources in infant as well as in adult nutrition. Thus, it is
an aim of the present invention to provide lipid preparations,
particularly cost effective preparations, with high levels of
cerebral-like lipids for advanced infant nutrition and for use in
dietary supplements, functional foods and pharmaceutical
compositions for promoting brain health.
[0034] WO 2005/051091 describes lipid preparations mimicking the
polar lipid composition of human breast milk fat (HMF), which
includes glycerophospholipids such as PC, PE, PS, and PI as well as
other polar lipids, such as sphingomyelin. These lipid preparations
are essentially obtained from mixtures of vegetable-derived
phospholipids, preferably soybean, as well as structured
phospholipids, such as trans-phosphatidylated lecithins. Other
lipid preparations mimicking the polar lipids of HMF described in
said publication comprise bovine milk-derived sphingomyelin. In
that earlier application, the inventors used pure bovine milk
sphingomyelin, obtainable as an analytical standard or research
chemical, which is not particularly suitable for use in infant
nutrition or dietary supplements due to its high cost and extremely
low availability, as mentioned previously.
[0035] Thus, it is a purpose of the current invention to provide
polar lipid preparations mimicking the polar lipids of HMF,
optionally comprising SM, wherein the source of said polar lipids
is a natural non-brain lipid source.
[0036] It is another object of the present invention to provide a
dietary supplement which guarantees the sufficient and recommended
intake of phospholipids, especially of PS and sphingomyelin, in the
form of a mimetic substitute of the phospholipids from human breast
milk lipid, aimed especially for infants and young children
consumption, as well as pregnant women. Other uses and objects of
the invention will become apparent as the description proceeds.
SUMMARY OF THE INVENTION
[0037] The present invention relates to a lipid preparation which
comprises a mixture of polar lipids, particularly
glycerophospholipids being phosphatidyl-choline (PC),
phosphatidylethanolamine (PE), phosphatidylserine (PS) and
phosphatidyl-inositol (PI), and optionally or non-optionally
comprising sphingolipids or a precursor or metabolite thereof,
preferably sphingomyelin, wherein the level of each of said PC, PE,
PS and PI is at least 1% w/w, and wherein said polar lipids are
derived from a natural non-brain animal lipid source.
[0038] The lipid preparation of any one of claims 1 and 2, wherein
the quantitative ratio between said glycerophospholipids is
substantially equivalent to said ratio in naturally occurring human
milk fat (HMF), at any desired stage of lactation.
[0039] In one embodiment, the lipid preparation of the invention
comprises a sphingolipid or a precursor or a metabolite thereof,
preferably sphingomyelin, wherein the quantitative ratio between
the glycerophospholipids and the sphingolipid in said preparation
is essentially equivalent to their corresponding ratio in said
naturally occurring HMF, at any desired stage of lactation.
[0040] Thus, the ratio between the polar lipids in the lipid
preparation of the invention is PC>PE>PS>PI, or
SM>PC>PE>PS>PI, or even SM=PC>PE>PS>PI.
[0041] In another embodiment said polar lipids are obtained from
said natural non-brain animal lipid source by fractionation and/or
extraction, or by a method that does not involve
transphosphatidylation.
[0042] Advantageously, the polar lipids comprising in said lipid
mixture are obtained from said natural non-brain animal lipid
source by fractionation and/or extraction, particularly by methods
which do not involve transphosphatidylation.
[0043] The quantitative ratio between said glycerophospholipids may
essentially mimic their corresponding ratio in naturally occurring
human milk fat (HMF), at any desired stage of lactation. Likewise,
when the preparation contains sphingomyelin (SM) or a precursor or
a metabolite thereof, the quantitative ratio between the
glycerophospholipids and the sphingomyelin in said preparation may
essentially mimic their corresponding ratio in said naturally
occurring HMF, at any desired stage of lactation.
[0044] In a particular embodiment, the invention relates to a lipid
preparation which is rich in PS, containing about 5-99% w/w PS,
preferably about 5-90% w/w, more preferably about 7-60% w/w, most
preferably about 7-25% w/w, wherein the PS is obtained from a
natural non-brain animal lipid source by methods which do not
involve transphosphatidylation, and wherein said lipid source is
preferably a marine source, milk or eggs, more preferably bovine
milk. This preparation may comprise additional polar lipids,
preferably PC, more preferably a mixture of PC, PE, PI, most
preferably a mixture of PC, PE, PI, and SM. The PS may be
characterized by a fatty acid profile characteristic of the animal
source from which it is derived, preferably the fatty acid profile
of bovine milk PS.
[0045] In another particular embodiment, the invention relates to a
lipid preparation which is rich in SM, containing about 5-99% w/w
SM, preferably about 5-90% w/w, more preferably about 10-60% w/w,
most preferably about 10-50% w/w, wherein the SM is derived from a
natural non-brain animal lipid source. The SM may be characterized
by a fatty acid profile characteristic of said animal source,
preferably the fatty acid profile of bovine milk SM.
[0046] Still further, the invention relates to a lipid preparation
which is rich in both PS and SM, containing about 5-99% w/w PS and
SM, preferably about 5-90% w/w, more preferably about 7-60% w/w,
most preferably about 7-50% w/w, wherein the PS and SM are derived
from a natural non-brain animal lipid source by a method which does
not involve transphosphatidylation, said lipid source being
preferably milk or eggs, more preferably bovine milk.
[0047] In a further aspect, the invention relates to a process for
the preparation of a lipid preparation which comprises a mixture of
polar lipids, particularly glycerophospholipids being
phosphatidyl-choline (PC), phosphatidyl-ethanolamine (PE),
phosphatidylserine (PS) and phosphatidyl-inositol (PI), and
optionally or not optionally comprising sphingolipids or a
precursor or metabolite thereof, preferably sphingomyelin, from a
natural non-brain animal lipid source, comprising isolating said
lipid mixture from said natural non-brain animal lipid source by
methods which do not involve transphosphatidylation.
[0048] A specific process for the preparation of the lipid
preparation of the invention, as described herein, comprises the
steps of:
[0049] (a) providing a natural non-brain animal lipid source which
has a substantially low content of polar lipids;
[0050] (b) removing non-lipid material from said lipid source,
dispersing the lipids, preferably with agitation, in a suitable
organic solvent or a mixture of organic solvents;
[0051] (c) separating the dissolved lipid fraction obtained in step
(b) and removing the organic solvent therefrom to give a lipid
fraction;
[0052] (d) de-oiling the lipid fraction obtained in step (c) at
least once to remove any non-polar lipids; and
[0053] (e) filtering and drying the polar lipids obtained in step
(d).
[0054] This process may further optionally comprise a step of
treatment of the lipid source with an aqueous medium, either before
or after said step (b).
[0055] Suitable organic solvents may be mixtures of a polar organic
solvent, preferably a primary alcohol, particularly ethanol, and a
non-polar solvent, preferably a hydrocarbon, particularly n-hexane.
The organic solvent may optionally contain water.
[0056] The process of the invention may be carried out at a
temperature of 15-100.degree. C., preferably 25-80.degree. C., more
preferably 30-60.degree. C.
[0057] The natural non-brain animal lipid source comprises at least
one lipid source, derived preferably from a marine source, more
preferably from an animal origin, most preferably from bovine milk
fat or from poultry eggs. The lipid source may be a mixture of
lipid sources.
[0058] The natural lipid source is preferably derived from bovine
milk, and may contain up to 5% w/w, preferably 10% w/w, more
preferably 25% w/w, even more preferably 35% w/w of total lipids,
in addition to other constituents including proteins and
carbohydrates, and wherein about 20%, preferably 30%, more
preferably 50%, most preferably 70% of said total lipids are polar
lipids.
[0059] In a specific embodiment the natural lipid source contains
about 0.1-10% w/w PC, about 0.1-5% w/w PE, about 0.1-5% w/w PS,
about 0.1-5% w/w PI and optionally contains sphingomyelin,
preferably at a level of about 0.1-5% w/w. More specifically, the
lipid source comprises about 2.6% w/w SM, about 4% w/w PC, about
3.2% w/w PE, about 1.6% w/w PS and about 0.9% w/w PI.
[0060] The lipid preparation resulting of any of the processes
described herein is characterized by a ratio of polar lipids of
PC>PE>PS>PI, or SM>PC>PE>PS>PI or
SM=PC>PE>PS>PI.
[0061] The present invention further provides a method for the
enrichment of SM in the lipid preparation obtained by any one of
the processes described herein, said method comprising further
subjecting said preparation to a step of any one of alkaline
hydrolysis, enzymatic hydrolysis, preparative chromatography or
polar lipid extraction.
[0062] In another embodiment the invention relates to a dietary
supplement or nutrient which comprises a lipid preparation of the
invention. The lipid preparation may be comprised in emulsified or
dispersed form, preferably in the form of an essentially aqueous
emulsion or dispersion or in dry form.
[0063] The invention also relates to a method for preparing a
dietary supplement of the invention, by admixing a lipid
preparation of the invention with at least one of additives,
emulsifiers or carriers. This method may further comprises admixing
the lipid preparation with an aqueous liquid medium, said dietary
supplement essentially being in an aqueous liquid form. The method
may further comprise dispersing, preferably dissolving the lipid
preparation in an organic medium, preferably an oil conventionally
used in infant formulas, particularly an oil which mimics HMF.
Still further, the method may comprise spray-drying the liquid
dietary supplement, to provide the dietary supplement in powder
form.
[0064] The lipid mixtures and preparations of the invention and the
dietary supplements or nutrients comprising them may be used as an
ingredient of a lipid constituent of infant formulas or as an
ingredient of infant formulas.
[0065] Still further, the lipid mixtures and preparations' of the
invention and the dietary supplements, nutrients or food articles
comprising them may be used in the enhancement of infants and/or
children development, particularly cognitive development and/or in
the enhancement of vision development.
[0066] A specific food article in accordance with the invention is
an infant formula, comprising the lipid preparation or mixture or a
dietary supplement comprising the same.
[0067] The invention also relates to a process for the preparation
of PS derived from a natural non-brain animal lipid source,
comprising the step of isolating PS from the polar lipid fraction
obtained by the process of the invention. The said natural
non-brain animal lipid source preferably comprises at least one
lipid source derived preferably from a marine source, more
preferably from an animal origin, most preferably from bovine fat
or from poultry eggs.
[0068] The invention further relates to a process for the
preparation of SM derived from a natural non-brain animal lipid
source, comprising the step of isolating SM from the polar lipid
fraction obtained by the process of the invention. The said natural
non-brain animal lipid source preferably comprises at least one
lipid source derived preferably from a marine source, more
preferably from an animal origin, most preferably from bovine milk
fat or from poultry eggs.
[0069] In a further aspect, the invention relates to the use of the
said PS, SM or PS/SM-rich preparations of the invention in the
improvement of cognitive functions, particularly memory,
concentration, attention and learning capabilities.
[0070] Furthermore, the invention relates to use of the said PS, SM
or PS/SM-rich preparations of the invention in the treatment of
brain-related illnesses or disorders or for improving cognitive
functions, e.g. mood-, memory-, stress-, or age-related disorders
and diseases such as dementia and Alzheimer's disease, as well as
memory loss and problems of concentration and attention and
learning capabilities.
[0071] In yet a further embodiment, the invention relates to use of
the said SM or PS/SM-rich preparations of the invention for
treating, preventing or ameliorating myelin-related disorders or
diseases, particularly de-myelination related disorders, such as
for example MS. These lipid preparations may also be used for
supporting and enhancing in a subject, particularly infants and
children, the normal or improved development of myelin sheath and
other sphingomyelin-related tissues.
DETAILED DESCRIPTION OF THE INVENTION
[0072] In a first aspect, the present invention provides a lipid
preparation comprising a mixture of polar lipids, particularly
glycerophospholipids being phosphatidylcholine (PC),
phosphatidylethanolamine (PE), phosphatidylserine (PS) and
phosphatidyl-inositol (PI), and optionally comprising a
sphingolipid or a precursor or metabolite thereof, particularly
sphingomyelin, wherein the level of each of said PC, PE, PS and PI
is at least 1% w/w, and wherein said polar lipids are derived from
a natural non-brain animal lipid source.
[0073] In said preparations, the ratio between polar lipids is:
SM>PC>PE>PS>PI or SM-PC>PE>PS>PI. Thus, the
ratio between SM and PC is either >1 or 1:1, preferably 1.1,
more preferably 1.3. Alternatively, the ratio between SM and PC is
1.5, preferably 2.
[0074] The lipid preparation of the invention is obtained by the
treatment, processing and fractionation of natural extracted polar
lipids mixtures, preferably containing glycerophospholipids, most
preferably also containing sphingomyelin. In a preferred embodiment
said natural lipids are animal sourced, preferably from milk of
farm animals, particularly bovine, and/or from poultry eggs. In a
further embodiment the natural lipid mixture may be derived from a
vegetal source, preferably containing sphingomyelin or a precursor
or metabolite of sphingomyelin.
[0075] In a particularly preferred embodiment the lipid preparation
of the invention may be prepared from a commercial bovine milk
preparation which contains low, and even extremely low levels of
the polar lipids (e.g. the mixture described in Table 2) and where
the lipids differ in their internal ratio from the
SM>PC>PE>PS>PI ratio that is found in HMF (Table 1
above).
TABLE-US-00002 TABLE 2 Commercial milk lipids Commercial milk
lipids Lipid class (weight %) (% from total polar lipids) SM 2.6
21.1 PC 4 32.5 PE 3.2 26.0 PS 1.6 13.0 PI 0.9 7.4 Total 12.3
100.00
[0076] In accordance with the present invention, the inventors have
processed and selectively extracted the polar lipids of the raw
bovine milk-derived starting mixture, thereby increasing the levels
of the individual lipids, and obtaining a preparation which is
suitable for use as a dietary supplement or as an infant nutrition
supplement. Importantly, the relative level of sphingomyelin has
been increased, up to a level similar or even higher than the level
of PC, thus making the lipid preparation of the invention even more
similar to HMF polar lipids. Additionally, the level of PE was
lowered, again making it comparable to the relative level of PE
found in HMF. Table 3 gives an example of a polar lipid preparation
of the invention, and details the relative levels of the polar
lipids, namely SM, PC, PE, PS, and PI, as compared to HMF lipids.
The SM>PC>PE>PS>PI ratio that is found in HMF, was
obtained in the lipid preparation of the invention (table 3), even
though the raw bovine milk-derived starting mixture has a different
ratio (PC>PE>SM>PS>PI).
TABLE-US-00003 TABLE 3 HMF lipids Lipid preparation of invention (%
from total SM, Lipid class (% from total SM, PC, PE, PS, PI) PC,
PE, PS, PI) SM 29.7 37.5 PC 27.9 28 PE 19.6 19.5 PS 13.3 9 PI 9.5 6
Total 100 100
[0077] The glycerophospholipids composition of the lipid
preparation of the invention is also comparable and mimetic to HMF
glycerophospholipids, as can be seen in the example presented in
Table 4.
TABLE-US-00004 TABLE 4 Lipid Lipid preparation of invention Human
Milk Fat class (% from total PC, PE, PS, PI) (% from total PC, PE,
PS, PI) PC 39.7 45.4 PE 27.8 30.8 PS 18.9 14.1 PI 13.5 9.7 Total
100 100
[0078] The composition of a specific lipid preparation of the
invention is described in Table 5. As can be seen, the preparation
consists mainly of high levels of HMF-like polar lipids, in
contrast to the low levels of polar lipids found in the starting
mixture. Thus, the preparation of the invention achieves levels of
polar lipids which are equivalent to the levels found in HMF. These
high levels permit the use of the preparations of the invention in
a variety of infant nutritional articles, dietary supplements,
functional foods, and pharmaceutical compositions.
[0079] It is important to note that the preparation of the
invention is man-made, and its constituents, even when derived from
natural sources, undergo processing/structuring before use in the
preparation of the invention. Alternatively, these constituents are
chemically or enzymatically synthesized. Therefore, the
constituents that are comprised in the preparation of the invention
contain at least traces of substances which are endogenous to the
lipid sources from which they are derived (such traces being edible
and physiologically compatible). Traces of this kind are absent
from human milk (and from HMF), which makes the preparation of the
invention similar, but not identical, to HMF.
[0080] In addition, HMF is the nomenclature used for the full lipid
fraction of human milk. It definitely contains the constituents
(the phospholipids) specifically mentioned herein, while also
containing other lipid constituents which are not present in the
lipid preparation of the invention, such as e.g. fat-soluble
vitamins. Thus, the most preferred embodiment of the lipid
preparation of the present invention comprises a combination of
phospholipids whose ratio is comparable to that of HMF
(SM>PC>PE>PS>PI or SM=PC>PE>PS>PI), and
further comprises traces of substances from lipid sources from
which said phospholipids are derived, while lacking other lipid
constituents which are present in HMF.
TABLE-US-00005 TABLE 5 Lipid class Lipid preparation of invention
(% w/w) SM 22.7 PC 21.4 PE 15.0 PS 10.2 PI 7.3 Total 76.6
[0081] The lipid preparation of the invention can be further
manipulated, in order to become even more similar to HMF, by
increasing the level of SM relative to the level of the
glycerophospholipids. This can be achieved by using one of the
following methods:
[0082] 1. Taking advantage of the known stability of SM to alkaline
hydrolysis, the lipid preparation of, for example, Table 5, is
subjected to alkaline aqueous conditions. The glycerophospholipids
undergo hydrolysis, resulting in the cleavage of their fatty acids
and the production of free fatty acids and partially or fully
hydrolyzed glycerophospholipids which are highly water soluble.
Thus, the non-hydrolyzed, relatively hydrophobic SM can be easily
separated and obtained by extraction to an organic or fatty media.
The obtained SM can be further combined with the preparation of
Table 5 (or other preparation of the invention), to obtain a lipid
preparation in which the relative level of SM is higher and closer
to that of HMF (around 38% of total polar lipids). Optionally, the
alkaline hydrolysis is conducted partially, hydrolyzing only part
of the glycerophospholipids. Thus, the hydrolysis process can be
controlled in a way that the non-hydrolyzed glycerophospholipids
and SM yield a lipid preparation which resembles the polar lipids
of HMF even more closely that of the preparation of Table 5;
[0083] 2. Utilizing selective enzymatic hydrolysis of
glycerophospholipids, by applying a glycerophospholipid selective
lipase to the polar lipid mixture of the invention (the preparation
of table 5 for example). By using one or more of the following
enzymes: 1:3 lipase, phospholipase A1 (PLA1) or phospholipase A2
(PLA2), the glycerophospholipids undergo hydrolysis of one or two
of the fatty acids. The partially or fully hydrolyzed fatty acids
can be separated from the SM as described following alkaline
treatment. The obtained SM can be further combined with the
preparation of Table 5 (or similar preparation). Alternatively, the
selective enzymatic hydrolysis can be conducted partially, by
applying less enzyme and/or having a shorter reaction time. This
way, the remaining glycerophospholipids and SM yield a lipid
preparation which resembles the polar lipids of HMF even more
closely than the original preparation;
[0084] 3. Using the different polarities between the different
components of the preparation of the invention
(SM>PC>PS>PI>PE) and conducting purification by means
of normal phase preparative chromatography. The obtained SM can be
further combined with the preparation of Table 5 (or similar
preparation), to obtain a lipid preparation in which the relative
level of SM is higher and closer to that of HMF;
[0085] 4. Conducting a polar lipid extraction from the raw material
(composed of proteins/sugars/lipids) through a selective solvent
system that increases the content of SM in either the extract or
the fraction of fat left and not extracted from raw material.
Examples of extraction solvents are any alcohol or a combination of
an alcohol and water. In this way, the SM, as the most polar
component, is enriched relative to its concentration in the raw
material. As previously described, the obtained SM may be combined
with the original preparation (of table 5 for example) in order to
obtain a lipid preparation richer in SM and more similar to
HMF.
[0086] Thus, the present invention provides a lipid preparation
which is obtained without mixing several lipid sources, but rather
by the fractionation and treatment of natural polar lipids
preparations, containing glycerophospholipids, preferably PC, PE,
PS, and PI, and optionally comprising sphingomyelin (or precursors
and/or metabolites of sphingomyelin). In accordance with the
present invention, the polar lipids mixtures are obtained from raw
lipid preparations extracted from natural sources, preferably
marine source, more preferably animal source, and even more
preferably bovine milk or poultry eggs. Milk of other mammals or
eggs of other domesticated or wild birds are also suitable as a
source. Furthermore, vegetal sources can be used whenever they
contain glycerophospholipids at a relative ratio that permits their
use without further mixing with other lipid sources, and/or the
lipids of vegetal sources include PS and/or sphingomyelin (or
precursors or metabolites of sphingomyelin). Examples or vegetal
sources are soybeans, sweet potatoes and peanuts.
[0087] In preferred embodiments, the lipid preparation in
accordance with the invention comprises above 1% w/w of each of PC,
PE, PS and PI, and optionally above 1% w/w of sphingomyelin,
preferably above 3% w/w of each of PC, PE, PS and PI, and
optionally above 3% w/w of sphingomyelin, more preferably above 5%
w/w of each of PC, PE, PS and PI, and optionally above, 5% w/w of
sphingomyelin, most preferably above 7% w/w of each of PC, PE, PS
and PI, and optionally above 7% w/w of sphingomyelin.
[0088] In a further preferred embodiment the level of PC is above
5% w/w, more preferably above 10% w/w, most preferably above 15%
w/w. The level of PC is preferably about 5-50% w/w, more preferably
about 8-40% w/w, most preferably 10-35% w/w.
[0089] In a preferred embodiment the level of PS is above 2% w/w,
more preferably above 4% w/w, most preferably above 7% w/w. The
level of PS is preferably about 2-50% w/w, more preferably about
4-40% w/w, most preferably about 5-25% w/w.
[0090] In a preferred embodiment the level of PE is above 4% w/w,
more preferably above 7% w/w, most preferably above 10% w/w. The
level of PE is preferably about 3-50% w/w, more preferably about
5-40% w/w, most preferably about 7-30% w/w.
[0091] In a preferred embodiment the level of PI is about 2-50%
w/w, more preferably about 3-40% w/w, most preferably about 4-20%
w/w.
[0092] In a preferred embodiment the level of SM is above 3% w/w,
more preferably above 5% w/w, more preferably above 10% w/w, most
preferably above 15% w/w. The level of SM is preferably about 5-50%
w/w, more preferably about 10-40% w/w, most preferably about 15-35%
w/w.
[0093] In a specifically preferred embodiment the relative levels
of the polar lipids (SM, PC, PE, PS, PI) mimic the relative levels
of these lipids in HMF in which SM>PC>PE>PS>PI.
Alternatively, said relative levels are SM=PC>PE>PS>PI. In
more preferred embodiments the relative level of SM of total polar
lipids (SM, PC, PE, PS, PI) is between 5-60%, more preferably
10-50%, even more preferably 20-40%, most preferably 30-35%.
[0094] In other specifically preferred embodiments the relative
level of PC of total polar lipids (SM, PC, PE, PS, PI) is between
5-60%, more preferably 10-50%, more preferably 20-40%, even most
preferably 25-35%.
[0095] In other specifically preferred embodiments the relative
level of PE of total polar lipids (SM, PC, PE, PS, PI) is between
5-50%, more preferably 10-40%, most preferably 23-30%.
[0096] In other preferred embodiments the relative level of PS of
total polar lipids (SM, PC, PE, PS, PI) is between 3-40%, more
preferably 5-35%, even more preferably 7-25%, most preferably
10-20%.
[0097] In other preferred embodiments the relative level of PI of
total polar lipids (SM, PC, PE, PS, PI) is between 2-40%, more
preferably 3-35%, most preferably 5-20%.
[0098] In another preferred embodiment the relative levels of the
glycero-polar lipids (PC, PE, PS, PI) mimic the relative levels of
these lipids in HMF in which PC>PE>PS>PI. In a more
preferred embodiments the relative level of PC out of total
glycero-polar lipids (PC, PE, PS, PI) is between 5-60%, more
preferably 10-55%, even more preferably 20-50%, most preferably
35-45%.
[0099] In another preferred embodiments the relative level of PE
out of total glycerol polar lipids (PC, PE, PS, PI) is between
5-60%, more preferably 10-50%, even more preferably 15-45%, most
preferably 20-35%.
[0100] In another preferred embodiment the relative level of PS out
of total glycero-polar lipids (PC, PE, PS, PI) is between 3-50%,
more preferably 5-45%, even more preferably 7-35%, most preferably
15-25%.
[0101] In yet another preferred embodiment the relative level of PI
out of total glycero-polar lipids (PC, PE, PS, PI) is between
3-50%, more preferably 5-40%, most preferably 7-30%.
[0102] The preparation of the invention comprises high levels of
PS, preferably above 2% w/w, preferably above 5% w/w, more
preferably above 8% w/w, most preferably above 10% w/w. The said PS
is of natural non-brain sources, preferably from milk or eggs,
preferably bovine milk or poultry eggs. It is to be noted that the
present invention can utilize a natural source of PS, comparable to
soybean transphosphatidylated PS, but from a safe natural source.
The preparation of the invention can serve as a good source of PS
obtainable by extraction and not by transphosphatidylation. This PS
can be used for the same purposes as commercial soybean
transphosphatidylated PS, particularly as a dietary supplement or
in functional foods or pharmaceutical preparations, for obtaining
the health benefits associated with PS supplementation.
[0103] The preparation of the invention is also characterized by
comprising high levels of SM, preferably above 3% w/w, more
preferably above 5% w/w more preferably above 10% w/w, and most
preferably above 15% w/w. The SM is of natural non-brain sources,
preferably from milk or eggs, more preferably bovine milk or
poultry eggs. Importantly, the preparation of the invention serves
as an affordable source of SM, which is not from animal brain, and
in the form, cost and grades applicable to dietary supplements,
functional foods and/or pharmaceutical compositions, having
brain-related health benefits.
[0104] Still further, the invention relates to a combination of PS
and SM, for use in the treatment of brain related illnesses or
disorders or for improving cognitive functions (mood deterioration,
depression, stress, age-related disorders and diseases such as
dementia, Alzheimer's disease, memory loss, problems of
concentration and attention, learning capabilities, etc.).
[0105] The invention further relates to lipid preparations
comprising PS and SM, optionally together with other polar lipids,
which exhibit a synergistic beneficiary effect between the PS and
SM. Both PS and SM are structural components of different tissues
or parts of the human brain. PS is an important building block of
cell membranes of grey matter and of nerve cells while SM is
essential component of myelin sheaths. Thus the supplementation of
these two important lipid-based brain building blocks has an
increased positive effect on cognitive functions, by treating and
maintaining the structural integrity of two brain systems
simultaneously, in a synergistic, not merely additive manner. These
compositions of PS and SM may contain above 1%, preferably 5-90%
w/w, more preferably 5-60% w/w, most preferably 5-40% w/w of each
of the PS and SM.
[0106] The invention specifically relates to a process of preparing
the lipid preparation of the invention. This process comprises the
steps of providing a suitable raw lipid source, removing from the
raw lipid source non-lipid material by selectively dissolving the
lipids in appropriate organic solvent or mixture of organic
solvents.
[0107] The raw lipid source can be a mixture of different lipid
sources, for example bovine milk combined with egg lecithin.
Alternatively, each of the different sources may be treated
independently by the process of the invention, and the final lipid
mixtures obtained can be mixed to give a lipid preparation
comprising polar lipids from varied sources.
[0108] In a preferred embodiment the lipid source is dispersed in a
mixture of non-polar and a polar organic solvent/s or polar solvent
alone. Preferred polar organic solvents are alcohols, preferably
primary alcohols, more preferably ethanol, and preferred non-polar
solvents are hydrocarbons, most preferably hexane. The solvent or
solvent system optionally includes water, or alternatively the
treatment with the organic solvents is preceded or followed by a
treatment with an aqueous medium. The above mentioned treatments
include dispersing the source of lipids in said media using
agitation. The treatment is carried out at room temperature, or
alternatively under cooling or heating conditions. Preferably the
treatment, removal of non-lipid material, is carried out at
temperatures where this removal is optimal and minimal amounts of
lipids are removed. Preferably the process occurs at elevated
temperatures, preferably 15-100.degree. C., more preferably
25-80.degree. C., most preferably 30-60.degree. C. The non-lipid
material is filtered off and is optionally further treated with
said solvent systems to ensure minimal amounts of polar lipids are
removed.
[0109] The organic solvents containing the lipids are removed by
conventional methods, preferably by evaporation under reduced
pressure. The lipid fraction is further de-oiled (removal of
non-polar lipids, mainly triglycerides) by conventional methods,
preferably, acetone or supercritical CO.sub.2. In case of acetone,
for example, the de-oiling step is repeated several times and the
resulting polar lipids are filtered and dried by conventional
methods, preferably under reduced pressure.
[0110] The preparation of the invention is suitable as a supplement
or additive for infant formulas, as a source of HMF-like polar
lipids, making said infant formulas more similar to human breast
milk, as well as in baby or toddler's foods, enriching their
nutritional content with HMF-like polar lipids.
[0111] The term "infant formula" as used herein encompasses infant
formulas (for newborn to 6 months old infants), follow-up formulas
(for 6-12 months old babies) and growing-up formulas (for 1-3 years
old children).
[0112] The term "infant formula" as used herein encompasses infant
formulas (for newborn to 6 months old infants), follow-up formulas
(for 6-12 months old babies) and growing-up formulas (for 1-3 years
old children).
[0113] The preparation of the invention can be further used to
mimic or create fat globules which mimic human milk fat
globules.
[0114] The preparation of the invention is also suitable in
supplementing the nutrition of pregnant women, either as a dietary
supplement, as an additive in functional foods, or as a
pharmaceutical preparation.
[0115] The preparation of the invention can be used for the
nutrition of pre-term infants as an additive in their nutrition, or
as a pharmaceutical preparation.
[0116] The preparation of the invention can be further used as an
active ingredient in dietary supplements, functional foods or
pharmaceutical preparations for improving the health of all human
beings, children, juveniles and young adults, adults, and
especially the elderly.
[0117] A specific aspect of the present invention is the use of the
preparation of the invention in improving the brain functions of
any of the above populations, specifically by improving their
cognitive functions, memory, alleviating stress, improving mood,
improving age-associated memory deterioration, fighting dementia,
preventing or treating Alzheimer's disease, etc.
[0118] The preparation of the invention may also be used for
treating, preventing or improving myelin-related disorders or
diseases, particularly de-myelination related disorders, such as
for example MS, upon its administration (or consumption) to
subjects in need.
[0119] The preparation of the invention is suitable for enabling
normal brain and cognitive development of infants, toddlers and
young children, through ensuring the normal or improved development
of the myelin sheath and other sphingomyelin-related tissues.
[0120] The preparation of pharmaceutical compositions is well known
in the art and has been described in many articles and textbooks,
see e.g., Gennaro A. R. ed. (1990) Remington's Pharmaceutical
Sciences, Mack Publishing Company, Easton, Pa., and especially
pages 1521-1712 therein.
[0121] Lastly, the present invention also provides methods of
treatment and/or improvement of cognitive functions, or
brain-related illnesses or disorders, for a subject in need, said
method comprising administering a therapeutically effective amount
of the lipid preparations provided in the present invention to said
subject. Said cognitive functions are memory, concentration,
attention and learning capabilities. Said disorders are mood-,
memory-, stress- or age-related neurological disorders, and
diseases such as dementia, Alzheimer's disease, memory loss,
problems of concentration and attention and learning
capabilities.
[0122] In addition, wherein said method of treatment utilizes the
lipid preparations of the invention comprising SM, said method is
suitable for subjects suffering from myelin-related disorders or
diseases, particularly de-myelination related disorders, such as
multiple sclerosis (MS) and leukodystrophies.
[0123] Usually, a "therapeutically effective amount" is determined
by the severity of the disease in conjunction with the preventive
or therapeutic objectives, the route of administration and the
patient's general condition (age, sex, weight and other
considerations known to the attending physician). The decision as
to the particular dosage to be employed (and the number of times to
be administered per day) is within the discretion of the physician,
and may be varied by titration of the dosage to the particular
circumstances of this invention to produce the desired therapeutic
effect.
[0124] The present invention further provides the use of the lipid
preparations described in the invention in the preparation of
pharmaceutical compositions for use in the treatment of the
above-described conditions.
[0125] Disclosed and described, it is to be understood that this
invention is not limited to the particular examples, process steps,
and materials disclosed herein as such process steps and materials
may vary somewhat. It is also to be understood that the terminology
used herein is used for the purpose of describing particular
embodiments only and not intended to be limiting since the scope of
the present invention will be limited only by the appended claims
and equivalents thereof.
[0126] It must be noted that, as used in this specification and the
appended claims, the singular forms "a", "an" and "the" include
plural referents unless the content clearly dictates otherwise.
[0127] Throughout this specification and the claims which follow,
unless the context requires otherwise, the word "comprise", and
variations such as "comprises" and "comprising", will be understood
to imply the inclusion of a stated integer or step or group of
integers or steps but not the exclusion of any other integer or
step or group of integers or steps.
[0128] The following Examples are representative of techniques
employed by the inventors in carrying out aspects of the present
invention. It should be appreciated that while these techniques are
exemplary of preferred embodiments for the practice of the
invention, those of skill in the art, in light of the present
disclosure, will recognize that numerous modifications can be made
without departing from the spirit and intended scope of the
invention.
EXAMPLES
Example 1
Preparation of a SM-Containing Polar Lipid Mixture
[0129] 50 gr of bovine milk preparation (Table 2) were added to 400
ml of hexane:ethanol (80:20) and mixed together at 40-45.degree. C.
After 2 hrs, the slurry was vacuum filtered and the cake
re-slurried under similar conditions and filtered again. The
combined solvent fractions were evaporated under reduced pressure
(<10 mbar). The lipid fraction obtained was further de-oiled
twice with acetone in a ration of 1:6 (w/volume) and the powder
obtained as sediments was filtered and dried under reduced pressure
(<10 mbar) in a vacuum dryer. 7 gr of dry powder were obtained
and analyzed to give the polar lipids distribution of Table 3
above.
* * * * *