WO2016043583A1

WO2016043583A1 - Reducing anxiety in progeny

Info

Publication number: WO2016043583A1
Application number: PCT/NL2015/050636
Authority: WO
Inventors: Johan Garssen; Aletta Desiree Kraneveld
Original assignee: Nutricia NV
Current assignee: Nutricia NV
Priority date: 2014-09-15
Filing date: 2015-09-15
Publication date: 2016-03-24
Anticipated expiration: 2017-03-15
Also published as: EP3193886A1; CN107106582A

Abstract

Anxiety is reduced in an infant that receives breast milk from a mother that has ingested non-digestible oligosaccharide.

Description

Reducing anxiety in progeny

Field of the invention

The present invention relates to the field of nutrition or nutritional supplements for lactating women, in particular nutrition that provides health benefits for the progeny of the pregnant women after birth thereof.

Background of the invention

The start of life is a stressful phase and from the moment it is born, an infant can be exposed to all kinds of factors that can influence the infant's state of mind. It can be expected that the mother of a newborn does anything to positively influence the way her baby feels.

With the increasing awareness of mothers that the diet has an influence on the quality of breast milk, there is also an increasing willingness of mothers that breastfeed their babies to rely more and more on nutritional support that would translate into beneficial effects in their babies.

Summary of the invention

In searching for factors that could be of help as a nutritional support for a breastfeeding or lactating mother to positively influence her baby, the present inventors now surprisingly found that by administering prebiotics to lactating mothers, their progeny displayed reduced anxiety. This was evidenced in a mouse model by a decrease in the vocalisations that were emitted by the progeny in the early stage of life when they were only breast-fed by their mothers, and where the mothers received a diet that was supplemented with prebiotics compared to progeny that were breast-fed by mothers that received a control diet without the prebiotics.

Detailed description of the invention

Thus, the present invention relates to a method for reducing anxiety in a breast-fed infant by administering a non-digestible oligosaccharide to the breastfeeding mother of the infant. In one embodiment according to the invention, the method for reducing anxiety is a non-medical method.

The invention can also be worded as the use of a non-digestible oligosaccharide in the manufacture of a composition for reducing anxiety in a breast-fed infant wherein the composition comprising a non-digestible oligosaccharide is administered to the breastfeeding mother of the infant.

The invention can also be worded as a non-digestible oligosaccharide for use in reducing anxiety in a breast-fed infant by administering non-digestible oligosaccharide to the breastfeeding mother of the infant.

Non-digestible oligosaccharides

The present method or use involves the administration of a non-digestible oligosaccharide. The non-digestible oligosaccharide in the method or use according to the present invention is preferably (i) not or only partially digested in the intestine by the action of acids or digestive enzymes present in the human upper digestive tract (small intestine and stomach) and (ii) fermented by the human intestinal flora, preferably fermentable into organic acids, preferably into lactic acid, butyrate, propionate and/or acetate. For example, sucrose, lactose, maltose and the maltodextrins are considered digestible.

Preferably, in the method or use according to the present invention, the non-digestible oligosaccharide has a DP from 2 to 250, more preferably from 2 to 60. The non- digestible oligosaccharide is at least one, preferably at least two selected from the group consisting of fructo-oligosaccharides, galacto-oligosaccharides, gluco- oligosaccharides, arabino-oligosaccharides, mannan-oligosaccharides, xylo- oligosaccharides, fuco-oligosaccharides, arabinogalacto-oligosaccharides, glucomanno- oligosaccharides, galactomanno-oligosaccharides sialic acid comprising oligosaccharides and uronic acid oligosaccharides. Preferably the present composition comprises fructo-oligosaccharides, galacto-oligosaccharides and/or galacturonic acid oligosaccharides, more preferably galacto-oligosaccharides, most preferably beta- galacto-oligosaccharides. The group of fructo-oligosaccharides includes inulin, the group of galacto-oligosaccharides includes transgalacto- oligosaccharides or beta- galacto-oligosaccharides, the group of gluco-oligosaccharides includes gentio-, nigero- and cyclodextrin-oligosaccharides and polydextrose, the group of arabinogalacto- oligosaccharides includes gum acacia, the group of galactomanno-oligosaccharides includes partially hydrolysed guar gum.

Preferably the non-digestible oligosaccharide comprises galacto-oligosaccharide. The galacto-oligosaccharide is preferably selected from the group consisting of beta- galacto-oligosaccharides, lacto-N-tetraose (LNT), lacto-N-neotetraose (neo-LNT), fucosyl-lactose, fucosylated LNT and fucosylated neo-LNT. In a particularly preferred embodiment the non-digestible oligosaccharide comprises beta-galacto- oligosaccharides. Beta-galacto-oligosaccharides as used in the present invention refers to oligosaccharides composed of over 50%, preferably over 65% galactose units based on monomeric subunits, with a degree of polymerization (DP) of 2 to 20, in which at least 50%, more preferably at least 75%, even more preferably at least 90%, of the galactose units are linked together via a beta-glycosidic linkage, preferably a beta- 1,4 glycosidic linkage. The average DP is preferably of 3 to 6. A glucose unit may be present at the reducing end of the chain of galactose units. Beta-galacto- oligosaccharides are sometimes also referred to as transgalacto-oligosacchariodes (TOS). A suitable source of beta-galacto-ologosaccharides is Vivinal®GOS (commercially available from Domo Ingredients, Netherlands). Other suitable sources are Oligomate® (Yakult), Cupoligo®, (Nissin) and Bimuno® (Classado).

On one embodiment, the non-digestible oligosaccharide comprises fructo- oligosaccharide. Fructo-oligosaccharide as used in the present invention refers to oligosaccharides composed of over 50%, preferably over 65 % fructose units based on monomeric subunits, in which at least 50%, more preferably at least 75%, even more preferably at least 90%, of the fructose units are linked together via a beta-glycosidic linkage, preferably a beta-2, 1 glycosidic linkage. A glucose unit may be present at the reducing end of the chain of fructose units. Preferably the fructo-oligosaccharide has a DP or average DP of 2 to 250, more preferably 2 to 100, even more preferably 10 to 60. Fructo-oligosaccaride comprises levan, hydrolysed levan, inulin, hydrolysed inulin, and synthesised fructo-oligosaccharides. In one embodiment, the non-digestible oligosaccharide comprises short chain fructo-oligosaccharides with an average degree of polymerization (DP) of 3 to 6, more preferably hydrolysed inulin or synthetic fructo- oligosaccharide. In one embodiment, preferably the non-digestible oligosaccharide comprises long chain fructo-oligosaccharides with an average DP above 20, such as Orafti HP® (Beneo, Belgium). Preferably the composition comprises both short chain and long chain fructo-oligosaccharides. Fructo-oligosaccharide suitable for use in the compositions is also readily commercially available, e.g. Orafti Synergy 1 ® (Beneo, Belgium). The non-digestible oligosaccharide used according to the present invention preferably comprises uronic acid oligosaccharides, more preferably galacturonic acid oligosaccharides. The term galacturonic acid oligosaccharide as used in the present invention refers to an oligosaccharide wherein at least 50% of the monosaccharide units present in the oligosaccharide is galacturonic acid. The galacturonic acid oligosaccharides used in the invention are preferably prepared from degradation of pectin, pectate, and/or polygalacturonic acid. Preferably the degraded pectin is prepared by hydrolysis and/or beta-elimination of fruit and/or vegetable pectins, more preferably apple, citrus and/or sugar beet pectin, even more preferably apple, citrus and/or sugar beet pectin degraded by at least one lyase. In a preferred embodiment, at least one of the terminal galacturonic acid units of the galacturonic acid oligosaccharide has a double bond. The double bond effectively protects against attachment of pathogenic bacteria to intestinal epithelial cells. Preferably one of the terminal galacturonic acid units comprises a C4-C5 double bond. The galacturonic acid oligosaccharide can be derivatised. The galacturonic acid oligosaccharide may be methoxylated and/or amidated. In one embodiment the galacturonic acid oligosaccharides are characterised by a degree of methoxylation above 20%, preferably above 50% even more preferably above 70%.

Thus, in one embodiment the non-digestible oligosaccharide in the method or use according to the present invention comprises at least beta-galacto-oligosaccharides. In one embodiment the non-digestible oligosaccharide in the method or use according to the present invention comprises at least short chain fructo-oligosaccharides and/or long chain fructo-oligosaccharides, preferably long chain fructo-oligosaccharides. In one embodiment the non-digestible oligosaccharide in the method or use according to the present invention comprises at least uronic acid oligosaccharides. In one embodiment the non-digestible oligosaccharide in the method or use according to the present invention comprises at least beta-galacto-oligosaccharides and at least short chain fructo-oligosaccharides or long chain fructo-oligosaccharides or both. In one embodiment the non-digestible oligosaccharide in the method or use according to the present invention comprises at least beta-galacto-oligosaccharides and at least uronic acid oligosaccharides. In one embodiment the non-digestible oligosaccharide in the method or use according to the present invention comprises at least short chain fructo- oligosaccharides and uronic acid oligosaccharides or long chain fructo-oligosaccharides and uronic acid oligosaccharides. In one embodiment the non-digestible oligosaccharide in the method or use according to the present invention comprises at least beta-galacto-oligosaccharides and short chain fructo-oligosaccharides and uronic acid oligosaccharides or at least beta-galacto-oligosaccharides and long chain fructo- oligosaccharides and uronic acid oligosaccharides.

In a preferred embodiment the non-digestible oligosaccharide in the method or use according to the present invention comprises a mixture of inulin and fructo- oligosaccharides. In a preferred embodiment the non-digestible oligosaccharide in the method or use according to the present invention comprises a mixture of galacto- oligosaccharides and fructo-oligosaccharides selected from the group consisting of short chain fructo-oligosaccharides and inulin, more preferably inulin.

In case a mixture of two different non-digestible oligosaccharides is used, preferably a mixture of galacto-oligosaccharides and fructo-oligosaccharide, preferably the weight ratio of the mixture of the two different non-digestible oligosaccharides is between 20 and 0.05, more preferably between 20 and 1. Preferably the non-digestible oligosaccharide in the method or use according to the present invention comprises galacto-oligosaccharides with a DP of 2-10 and fructo-oligosaccharides with a DP of 2- 60. In one embodiment, the non-digestible oligosaccharide comprises beta-galacto- oligosaccharide, fructo-oligosaccharide and a pectin degradation product. The weight ratio beta-galacto-oligosaccharide : fructo-oligosaccharide : pectin degradation product is preferably (20 to 2) : 1 : (1 to 20), more preferably (12 to 7) : 1 : (1 to 3). Preferably, the non-digestible oligosaccharide is administered to the lactating mother in a serving comprising from 0.05 to 50 grams non-digestible oligosaccharide, preferably from 0.1 to 25 grams, preferably from 0.5 to 15 grams, preferably from 1 to 10 grams, preferably from 1 to 5 grams. The present method or use preferably comprises the administration of a serving comprising from 0.05 to 50 grams galacto-oligosaccharide plus fructo-oligosaccharide , preferably from 0.1 to 25 grams, preferably from 0.5 to 15 grams, preferably from 1 to 10 grams, preferably from 1 to 5 grams galacto- oligosaccharide plus fructo-oligosaccharide. The present method or use preferably comprises the administration of a serving comprising from 0.05 to 50 grams galacto- oligosaccharide, preferably from 0.1 to 25 grams, preferably from 0.5 to 15 grams, preferably from 1 to 10 grams, preferably from 1 to 5 grams galacto-oligosaccharide.

The present method or use preferably comprises the administration of 0.05 to 50 grams non-digestible saccharide per day, preferably from 0.1 to 25 grams, preferably from 0.5 to 15 grams, preferably from 1 to 10 grams, preferably from 1 to 5 grams per day. The present method or use preferably comprises the administration of 0.05 to 50 grams galacto-oligosaccharide plus fructo-oligosaccharide per day, preferably from 0.1 to 25 grams, preferably from 0.5 to 15 grams, preferably from 1 to 10 grams, preferably from 1 to 5 grams galacto-oligosaccharide plus fructo-oligosaccharide per day. The present method or use preferably comprises the administration of 0.05 to 50 grams galacto- oligosaccharide per day, preferably from 0.1 to 25 grams, preferably from 0.5 to 15 grams, preferably from 1 to 10 gram, preferably from 1 to 5 grams galacto- oligosaccharide per day.

Breastfeeding women

In the context of the present invention a breastfeeding mother includes a woman who taps breast milk and feeds the tapped breast milk via a bottle to the infant. According to the method or use of the present invention, non-digestible oligosaccharide is administered to the breastfeeding mother of the infant. Preferably non-digestible oligosaccharide is administered to the breastfeeding mother from the day the infant is born. Preferably administration of non-digestible oligosaccharide to the breastfeeding mother is continued until the day the mother stops breastfeeding completely.

Infant

The present method or use is of benefit for healthy term born infants. In a preferred embodiment the present method or use is of benefit for infants that are prone to develop anxiety.

Composition

The non-digestible oligosaccharide may be administered to the lactating women in the form of a bar, a capsule, a tablet, a liquid, or a powder. Suitably a powder can be mixed with any type of food or beverage prior to intake thereof.

Preferably non-digestible oligosaccharide is in the form of a nutritional supplement for the lactating women. In one embodiment, the non-digestible oligosaccharide is a powder, packed in sachet comprising 1 to 10 g, more preferably 1.5 to 7 g, most preferably 2 to 5 g non-digestible oligosaccharide.

In the present method or use, preferably the non-digestible oligosaccharide is administered to a lactating woman in an amount from 0.5 to 50 g per day, preferably from 0.1 to 25 g per day, preferably from 0.5 to 15 g per day, preferably from 1 to 10 g per day, preferably from 1 to 5 g per day, preferably from 3 to 25 g per day, preferably from 6 to 12 g per day. Preferably, this daily dose is administered in one portion per day. Preferably, this daily dose is divided over 2 or 3 or 4 portions, which are consumed 2, 3 or 4 times per day, respectively.

Application

According to the present invention anxiety is reduced in an infant that receives breast milk from a mother that has ingested non-digestible oligosaccharide.

In this document and in its claims, the verb "to comprise" and its conjugations is used in its non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. In addition, reference to an element by the indefinite article "a" or "an" does not exclude the possibility that more than one of the element is present, unless the context clearly requires that there be one and only one of the elements. The indefinite article "a" or "an" thus usually means "at least one". Examples - Prebiotics in diet of lactating dam induces reduced anxiety in offspring

The effects of postnatal dietary intervention with short chain GOS and long chain FOS on, anxiety-like behavior was examined in healthy Balb/cByJ mice.

Pregnant Balb/cByJ mice (Charles River Laboratories, Maastricht, The Netherlands) were housed solitary in plastic cages containing standard chip bedding and free access to food and water. Balb/c mice are known as anti-social mice that display increased anxiety-like behavior compared to other murine models.

A 12h light dark cycle was followed (7 AM - 7 PM). The day of birth of the pups is postnatal day 0 (P0). From P0, n=6 dams were fed a diet enriched with 3% scGOS/lcFOS, and n=9 dams were fed the control diet (AIN93G).

On P21, male offspring was weaned. Two cages of five males per cage were randomly formed from the dams that were fed the GOS/FOS diet and two cages were randomly formed of 5 males per cage from the dams on the AIN93G diet. Furthermore, one cage of six males from dams on the control diet was formed, these mice functioned as interaction mice during the social interaction test. Offspring was provided with the same diet as the dam it came from, forming n=10 male mice on 3% GOS/FOS diet and a total of n=16 male mice on AIN93G diet.

Measurement of separation-induced ultrasonic vocalization

Ultrasonic distress vocalization of male offspring was measured on P10 to determine anxiety-like behavior. An ultrasonic vocalization testing cylinder with a lid in which a microphone (UltraSound Advice, London, United Kingdom) was placed, was connected to an amplifier and a computer with registration software (SonoTrack, Metris B.V.; Hoofddorp, The Netherlands). The aluminum bottom of the cylinder was connected to a Julabo F12-ED heating circulator (Sigma-Aldrich; Zwijndrecht, The Netherlands) to maintain the temperature at 19 °C. A pup was separated from the dam and its littermates and placed in the cylinder for three minutes, during which the number of calls, average time of calls and average wavelength of calls were measured. Dam and littermates were housed in the same room during this test. Statistical methods

Differences between groups were statistically analyzed with an unpaired two-tailed Students t-test. When not normally distributed, data was transformed using the natural logarithm before statistical analysis. Data are shown as average ± SEM, using GraphPad Prism, version 6. Results were considered statistically significant when P < 0.05.

Results

Pups lactated by a dam fed with scGOS/lcFOS diet show a reduction in ultrasonic vocalizations

On P10, ultrasonic distress vocalization was measured to assess anxiety-like behavior. Male pups were individually separated from the dam for 3 minutes, during which the amount of UV calls, average time of a call and the average frequency of a call were measured. The number of calls was significantly reduced in the pups that were lactated by dams on a scGOS/lcFOS diet, compared to mice that were lactated by dams on the control diet, (P = 0.040).

Furthermore, there is a trend that shows a reduction in the average time a call lasts in pups that were lactated by a dam on scGOS/lcFOS diet compared to control animals (P = 0.065).

No significant difference can be seen in the average frequency (Hz) of a call emitted by the different pups.

The results are summarized in table 1 Table 1 : Pups lactated by a dam fed with scGOS/lcFOS diet show a reduction in ultrasonic vocalizations

Control diet: AIN93G These results are indicative that a prebiotic diet during the lactation phase reduces anxiety in the offspring.

Claims

1. Use of a non-digestible oligosaccharide in the manufacture of a composition for reducing anxiety in a breast-fed infant wherein the composition comprising a non- digestible oligosaccharide is administered to the breastfeeding mother of the infant.

2. The use according to claim 1 wherein the non-digestible oligosaccharide comprises galacto-oligosaccharides, preferably with an average degree of polymerisation from 3 to 6.

3. The use according to any one of the preceding claims wherein the non-digestible oligosaccharide comprises fructo-oligosaccharides, preferably with an average degree of polymerisation above 10, preferably above 20.

4. The use according to any one of the preceding claims wherein the non-digestible oligosaccharide comprises galacto-oligosaccharides and fructo-oligosaccharides, preferably in a weight ratio from 20 : 0.5

5. The use according to any one of the preceding claims, wherein the infant is prone to develop anxiety.

6. The use according to any one of the preceding claims wherein the composition is in the form of a nutritional supplement.

7. The use according to any one of the preceding claims wherein the non-digestible oligosaccharide is administered to the breastfeeding mother from the day the infant is born.

8. The use according to any one of the preceding claims wherein administration of the non-digestible oligosaccharide is continued until the day the mother stops breastfeeding completely. The use according to any one of the preceding claims, wherein the non-digestible oligosaccharide is administered to the breastfeeding mother in an amount from 1 to 12 g per day.

Non medical method for reducing anxiety in a breast-fed infant by administering a non-digestible oligosaccharide to the breastfeeding mother of the infant.