Human milk oligosaccharides for use in improving executive function

Abstract

Use of human milk oligosaccharides, or a nutritional composition comprising human milk oligosaccharides, for improving executive function in non-infant infants, for preventing and / or reducing the risk of suboptimal executive function in non-infant infants, for maintaining suboptimal executive function in non-infant infants, and / or for improving myelination to mature the prefrontal cortex region of the brain in non-infant infants, as well as a pack comprising a synthetic composition of HMO and at least 14 individual daily doses of an effective amount of at least one human milk oligosaccharide (HMO) for such use.

Description

[Technical field]

[0001] The present invention relates to the use of human milk oligosaccharides (HMOs) or compositions comprising HMOs for improving executive function in non-infant infants. The present invention further relates to HMOs or compositions comprising HMOs for use in the prevention or management of suboptimal executive function in non-infant infants. [Background technology]

[0002] Executive functions are the abilities that coordinate and integrate cognitive-perceptual processes related to time and space, and determine how well a subject recognizes, evaluates, and selects between various alternative options and strategies. They govern goal-directed behavior and have a fundamental role in the control of higher-order cognitive processes such as problem solving, reasoning, flexible thinking, and decision-making. They are central to cognitive development and learning (especially of new skills) and are strongly associated with social and intellectual / academic success and / or achievement.

[0003] In humans, the abilities associated with executive function do not develop all at once. Instead, they develop sequentially, with each ability building on previous skills. All executive functions interact with each other and influence how humans control their behavior to produce positive future outcomes. Typically, executive functions begin to develop by age 2 and are fully developed by age 30. Executive functions typically begin to decline after age 30, most commonly related to memory.

[0004] Executive functions are thought to be mediated by the prefrontal cortex. Generally, information is stored in the back of the brain, and humans use the prefrontal cortex to process that information so they can be socially effective and succeed in life. Executive functions require four main control circuits: Task-oriented circuits that control working memory, enabling us to execute plans, take the specific steps required to complete a task, and achieve our goals Timing-oriented circuits that organize the order in which tasks are completed and allow for processing of time series Emotion-oriented circuits that control emotions and moods · Consciousness-directed circuits that control emotions and self-awareness of experience.

[0005] Non-infant humans with poor executive function (or executive dysfunction - EFD) struggle to organize and control their behavior in ways that help them achieve long-term goals. In general, non-infant humans with EFD are more likely to be motivated to act by short-term goals rather than long-term goals. The result of poor executive function is usually less success in life.

[0006] Despite the very real impact of EFD, the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) does not recognize EFD as a specific mental medical condition. Instead, problems with executive function are a symptom of other neurological, mental health, and behavioral disorders. Thus, while there is no diagnosis of EFD per se, there are diagnoses of neurological, mental health, or behavioral disorders with symptoms of higher-order dysfunction. Typical symptoms are emotional or impulse control problems, problems with initiating, organizing, planning, or completing tasks, listening or attention problems, short-term memory problems, inability to multitask or balance tasks, socially inappropriate behavior, inability to learn from past outcomes, difficulties with problem solving, and difficulties learning or processing new information.

[0007] When symptoms become evident, various tests can be used to help evaluate a person's executive function, such as the Stroop task test to assess a person's mental control and selective attention, the trail making test, the clock drawing test, the verbal fluency test, and the card sorting test. Alternatively, if a specific psychiatric disorder is suspected, medical professionals may skip the executive function tests and instead compare the person's symptoms to standard diagnostic criteria for that disorder, or use an MRI scan to detect physical causes, such as stroke damage or a brain tumor.

[0008] Diagnosable conditions that can cause problems with executive functioning include attention-deficit hyperactivity disorder (ADHD), depression and anxiety disorders, bipolar disorder, schizophrenia, obsessive-compulsive disorder (OCD), autism, Alzheimer's disease, frontotemporal dementia, Lewy body dementia, Tourette's syndrome, post-traumatic stress disorder (PTSD), and traumatic brain injury. However, there can also be temporary causes of problems with executive functioning, such as fatigue, severe pain, stress, distracting environments, drug use, alcohol use, and severe boredom. In addition, individuals born prematurely or small for gestational age (SGA) appear to be at higher risk for problems with executive functioning. When problems or impairments in executive functioning are present, the type of treatment used depends on the diagnosed condition believed to be causing the problem or impairment. Treatment options include stimulant medications, antidepressants, antipsychotics, psychoeducation, occupational or speech therapy, and cognitive behavioral therapy (CBT). First-line treatment is usually occupational or speech therapy, psychotherapy, reading instruction, or CBT. When combined with medication to treat the diagnosed disorder, CBT can be very effective in treating executive function disorders such as inhibition, emotion control, time management, and planning problems in adults. However, CBT is less effective in children. Also, there are not always effective medications for the diagnosed disorder, or the medications have serious side effects that reduce patient compliance. Furthermore, there are very limited approaches to prevent executive function problems or disorders in non-infant humans at risk.

[0009] Thus, there remains a need for safe and well-tolerated means to prevent or manage executive function problems or disorders in non-infant humans. Summary of the Invention

[0010] Surprisingly, it has been discovered that human milk oligosaccharides may prevent executive function problems or disorders, improve executive function and allow better management of executive function problems or disorders in non-infant humans.

[0011] Thus, in a first aspect, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis, comprising: a) Improving executive function in non-infants; b) preventing and / or reducing the risk of suboptimal executive function in non-infant children; c) Managing suboptimal executive function in non-infants; and / or d) Improved myelination for maturation of the prefrontal cortex region of the brain in non-infant infants The present invention relates to human milk oligosaccharides (HMOs) or synthetic compositions containing HMOs for use in

[0012] Preferably, the synthetic composition comprises HMO in an amount of 0.5 g to 15 g; more preferably 1 g to 10 g. For example, the synthetic composition may comprise HMO in an amount of 2 g to 7 g.

[0013] The synthetic composition may include bifidobacteria; for example, Bifidobacterium longum and / or Bifidobacterium bifidum.

[0014] A second aspect of the present invention is a) Improving executive function in non-infants; b) preventing and / or reducing the risk of suboptimal executive function in non-infant children; c) Managing suboptimal executive function in non-infants; and / or d) Enhanced myelination for maturation of the prefrontal cortex region of the non-infant brain The present invention relates to a pack for use in a dietary supplement comprising at least 14 individual daily doses of an effective amount of at least one human milk oligosaccharide (HMO).

[0015] Preferably, each dose in the pack contains from about 0.5g to 15g, more preferably from 1g to 10g, even more preferably from 2g to 7g of human milk oligosaccharides.

[0016] The pack preferably contains at least about 21 days' doses; for example about 28 days' doses.

[0017] The pack may contain bifidobacteria; for example, Bifidobacterium longum and / or Bifidobacterium bifidum.

[0018] A third aspect of the invention provides a method for improving executive function in a non-infant, the method comprising administering to the non-infant an effective amount of at least one human milk oligosaccharide (HMO).

[0019] Preferably, the method is effective to improve the non-infant's brain power to perceive, evaluate, and / or select from various alternative options and / or strategies. Preferably, the method further comprises providing cognitive behavioral therapy to the non-infant.

[0020] A fourth aspect of the present invention provides a method for preventing and / or reducing the risk of suboptimal executive function in a non-infant, the method comprising administering to the non-infant an effective amount of at least one human milk oligosaccharide (HMO).

[0021] The HMO may be administered to a non-infant at risk of developing non-optimal executive function later in life. In such cases, the method further comprises identifying a non-infant at risk of developing non-optimal executive function later in life. The non-infant may be an older human who was born prematurely or small for gestational age, or a child aged 3 years or older. The HMO may also be administered to a non-infant who previously suffered from non-optimal executive function to prevent and / or reduce the risk of recurrence or the severity of recurrence.

[0022] A fifth aspect of the invention provides a method for managing sub-optimal executive function in a non-infant, comprising administering to the non-infant an effective amount of at least one human milk oligosaccharide (HMO).

[0023] Preferably, the method is effective in managing a decline in the ability of a non-infant to recognize, evaluate, and / or select between various alternative options and / or strategies, and is particularly effective in improving short-term and long-term memory, and thereby executive function, in a non-infant.

[0024] Preferably, the method further comprises providing cognitive behavioral therapy to the non-infant. In some embodiments, the HMO is administered to a non-infant prior to adulthood (i.e., before reaching age 18).

[0025] Preferably, a non-infant is administered about 0.5 g to 15 g of human milk oligosaccharides per day, more preferably 1 g to 10 g per day, and even more preferably 2 g to 7 g per day.

[0026] The invention also provides HMOs or compositions comprising HMOs for use in the manufacture of a composition for use in the prevention and / or treatment of sub-optimal executive function in a non-infant.

[0027] The non-infant may have one or more of attention deficit hyperactivity disorder (ADHD), Alzheimer's disease, depression, anxiety, bipolar disorder, schizophrenia, obsessive-compulsive disorder (OCD), autism, Alzheimer's disease, frontotemporal dementia, dementia with Lewy bodies, Tourette's syndrome, post-traumatic stress disorder (PTSD), traumatic brain injury, or vascular dementia.

[0028] The HMO can be a fucosylated neutral HMO, a non-fucosylated neutral HMO, and / or a sialylated HMO.

[0029] In certain embodiments, the fucosylated neutral HMO can be 2'-fucosyllactose (2'-FL) and / or difucosyllactose (3,2'-difucosyllactose, DFL).

[0030] In certain embodiments, the nonfucosylated neutral HMO can be LNT (lacto-N-tetraose), LNnT (lacto-N-neotetraose), or a combination thereof.

[0031] In certain embodiments, the sialylated oligosaccharides can be 3'-sialyllactose (3'-SL) and / or 6'-sialyllactose (6'-SL).

[0032] In the context of the present invention, particularly effective HMO combinations may be: -2'-FL, DFL, LNT, and LNnT, -3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL), or -2'-FL, DFL, LNT, LNnT, 3'-sialyllactose (3'-SL), and 6'-sialyllactose (6'-SL).

[0033] In a sixth aspect, the present invention provides a method for enhancing myelination for maturation of the prefrontal cortex region of the non-infant brain, comprising administering at least one sialylated human milk oligosaccharide (HMO) to the non-infant. The sialylated HMO is 3'-SL, 6'-SL, or a combination thereof. Additional HMOs may be administered to the non-infant.

[0034] Preferably, the non-infant is administered about 0.5 g to 15 g per day; more preferably 1 g to 10 g per day of HMO. For example, the non-infant may be administered 2 g to 7 g per day. Preferably, the non-infant is administered the HMO for a period of at least one week; more preferably at least two weeks.

[0035] The non-infant may be administered a high dose in the initial phase and a low dose in the maintenance phase. Preferably, the non-infant is administered the HMO for a period of at least 1 week during the initial phase; more preferably, for a period of at least 2 weeks. The non-infant may be administered the HMO for a period of at least 4 weeks during the maintenance phase; more preferably, for a period of at least 8 weeks. The dose administered in the initial phase is preferably about 3g to about 10g per day (e.g., about 4g to about 7.5g per day), and the dose administered in the maintenance phase is preferably about 2g to about 7.5g per day (e.g., about 2g to about 5g per day). [Brief description of the drawings]

[0036] [Figure 1] Performance scores of piglets exposed to preweaning diets supplemented with milks A, B, C, control milk, or fed by sows, calculated to assess general working memory. Different letters indicate significant differences (p<0.05). [Diagram 2] Performance scores of piglets exposed to preweaning diets supplemented with milks A, B, C, control milk, or fed by sows, calculated to assess reference memory. Different letters indicate significant differences (p<0.05). [Diagram 3] Performance scores of piglets exposed to preweaning diets supplemented with milks A, B, C, control milk, or fed by sows, calculated to assess working memory. Different letters indicate significant differences (p<0.05). [Figure 4] Myelin gene expression in prefrontal and hippocampal brain samples from pups and adult mice fed milk without 6'-SL, normalized to expression levels in mice fed milk containing 6'-SL. Asterisks indicate significant differences from controls (p<0.05). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0037] Detailed Description of the Invention Surprisingly, it has been discovered that oral or enteral administration of one or more human milk oligosaccharides (HMOs) to non-infant humans a) improves executive function in non-infant humans, b) prevents and / or reduces the risk of suboptimal executive function in non-infant humans, c) manages suboptimal executive function in non-infant humans, and / or d) improves myelination for maturation of the prefrontal cortex of the non-infant brain.

[0038] As used herein, the following terms have the following meanings:

[0039] "Effective amount" means that amount of an HMO sufficient to produce a desired result in a human. An effective amount can be administered in one or more doses sufficient to produce the desired result.

[0040] "Enteral administration" means any conventional form for delivering a composition to a patient that results in deposition of the composition in the digestive tract (such as the stomach). Methods of enteral administration include nasogastric or jejunal tubes, oral, sublingual, and rectal feeding.

[0041] "Executive function" refers to the ability to perceive, evaluate, and select from various alternative options and strategies. The term includes goal-directed behavior, planning, and / or cognitive flexibility.

[0042] "Non-infant human" or "non-infant" means a human over the age of 3. A non-infant human can be a child, a teenager, an adult, or a senior (over the age of 65).

[0043] "Human milk oligosaccharides" or "HMOs" refer to complex carbohydrates present in human breast milk (Urashima et al.: Milk Oligosaccharides. Nova Science Publisher (2011); Chen Adv. Carbohydr. Chem. Biochem. 72, 113 (2015)). HMOs have a core structure containing lactose at the reducing end, which can be extended by one or more β-N-acetyl-lactosaminyl and / or one or more β-lacto-N-biosyl units, and this core structure can be substituted by α-L-fucopyranosyl and / or α-N-acetyl-neuraminyl (sialyl) moieties. In this regard, non-acidic (or neutral) HMOs lack sialyl residues and acidic HMOs have at least one sialyl residue in their structure. Non-acidic (or neutral) HMOs can be fucosylated or non-fucosylated. Examples of such neutral nonfucosylated HMOs include lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), lacto-N-neohexaose (LNnH), para-lacto-N-neohexaose (pLNnH), para-lacto-N-hexaose (pLNH), and lacto-N-hexaose (LNH).Examples of neutral fucosylated HMOs include 2'-fucosyllactose (2'-FL), lacto-N-fucopentaose I (LNFP-I), lacto-N-difucohexaose I (LNDFH-I), 3-fucosyllactose (3-FL), difucosyllactose (DFL), lacto-N-fucopentaose II (LNFP-II), lacto-N-fucopentaose III (LNFP-III), lacto-N-difucohexaose III (LNDFH-III), fucosyl-lacto-N-hexaose II (FLNH-II), lacto-N-fucopentaose V (LNFP-V), lacto-N-fucopentaose VI (LNFP-VI), lacto-N-difucohexaose Fucosyl-lacto-N-hexaose I (LNDFH-II), fucosyl-lacto-N-hexaose I (FLNH-I), fucosyl-para-lacto-N-hexaose I (FpLNH-I), fucosyl-para-lacto-N-neohexaose II (F-pLNnH II), and fucosyl-lacto-N-neohexaose (FLNnH). Examples of acidic HMOs include 3'-sialyllactose (3'-SL), 6'-sialyllactose (6'-SL), 3-fucosyl-3'-sialyllactose (FSL), LST a, fucosyl-LST a (FLST a), LST b, fucosyl-LST b (FLST b), LST c, fucosyl-LST c (FLST c), sialyl-LNH (SLNH), sialyl-lacto-N-hexaose (SLNH), sialyl-lacto-N-neohexaose I (SLNH-I), sialyl-lacto-N-neohexaose II (SLNH-II), and disialyl-lacto-N-tetraose (DSLNT).

[0044] "Oral administration" refers to any conventional form for delivering a composition to a human via the mouth. Oral administration is thus a form of enteral administration.

[0045] "Synthetic composition" refers to an artificially produced composition, preferably a composition that contains at least one compound that is chemically and / or biologically produced ex vivo, for example, by chemical reaction, enzymatic reaction, or recombinant. In some embodiments, the synthetic composition may contain one or more compounds or ingredients other than HMO. Also, in some embodiments, the synthetic composition may contain one or more nutritionally or pharmacologic active ingredients that do not adversely affect the efficacy of HMO. Some non-limiting embodiments of the synthetic composition of the present invention are also described below.

[0046] "Small for gestational age (SGA)" refers to a child aged 3 years or older who is smaller than normal for gestational age at birth, most commonly defined as weight below the 10th percentile for gestational age. In some embodiments, SGA may be associated with intrauterine growth restriction (IUGR), a condition in which a fetus fails to reach its potential size.

[0047] "Low birth weight" means weight at birth less than 2500g. It therefore includes: - Children who weigh between 1,800 and 2,500 grams at birth (commonly referred to as "low birth weight" or LBW), - Children who weighed between 1000 and 1800g at birth (called "very low birth weight" or VLBW), - Babies who weighed less than 1000g at birth (called "extremely low birth weight" or ELBW).

[0048] "Prophylactic treatment" or "prevention" means a treatment performed or measures taken to reduce the risk of onset or recurrence of a disease, or its severity after onset.

[0049] "Prophylactically reducing the severity and / or recurrence of a symptom" means reducing the severity and / or recurrence of a symptom at a later time.

[0050] "Secondary prevention" refers to preventing the onset of a condition in a high-risk patient or preventing the recurrence of symptoms in a patient already suffering from a condition. A "high-risk" patient is an individual who is predisposed to developing a condition, such as, for example, someone with a family history of a condition.

[0051] "Manage" means addressing a medical condition or disease with the goal of improving or stabilizing outcome or addressing underlying nutritional needs in a non-infant. Thus, management includes dietary or nutritional management of a medical condition or disease by addressing the nutritional needs of a non-infant. "Manage" and "management" have grammatically corresponding meanings. Management includes the treatment of a non-infant.

[0052] "Treatment" means a remedy administered or action taken to reduce or eliminate the symptoms of a disease or condition.

[0053] In a first aspect of the invention there is provided an HMO, or a composition comprising an HMO, for use in enhancing executive function in a non-infant.

[0054] The HMOs may be fucosylated neutral HMOs, non-fucosylated neutral HMOs, sialylated HMOs, or any combination of any of the above.

[0055] In certain embodiments, the HMO is a fucosylated neutral HMO. In certain embodiments, the HMO is a non-fucosylated neutral HMO. In certain embodiments, the HMO is a sialylated HMO.

[0056] In certain embodiments, the HMOs are a mixture comprising, consisting essentially of, or consisting of one or more fucosylated neutral HMOs and one or more non-fucosylated neutral HMOs.

[0057] In certain embodiments, the HMOs are a mixture comprising, consisting essentially of, or consisting of one or more fucosylated neutral HMOs, one or more non-fucosylated neutral HMOs, and one or more sialylated HMOs.

[0058] In particular, the one or more fucosylated neutral HMOs are selected from 2'-FL, 3-FL, DFL, LNFP-I, LNFP-II, LNFP-III, LNFP-V, LNFP-VI, LNDFH-I, LNDFH-II, LNDFH-III, FLNH-I, FLNH-II, FLNnH, FpLNH-I, and F-pLNnH II, preferably 2'-FL, 3-FL, and DFL. Particularly useful fucosylated oligosaccharides can be 2'-fucosyllactose (2'-FL) and difucosyllactose (DFL).

[0059] Preferably, the one or more nonfucosylated neutral HMOs are selected from LNT, LNnT, LNH, LNnH, pLNH, and pLNnH, preferably LNnT and LNT.

[0060] The one or more sialylated HMOs are preferably selected from 3'-SL, 6'-SL, LST a, LST b, LST c, DS-LNT, S-LNH, and S-LNnH I, more preferably 3'-SL and 6'-SL. HMOs that contain both fucosylated and sialylated moieties, such as FSL, may also be used.

[0061] In certain embodiments, the HMO is selected from the group consisting of 2'-fucosyllactose (2'-FL), DFL, LNT, LNnT, sialyllactose, and any combination of the foregoing.

[0062] In certain embodiments, the HMO is a combination of 2'-FL, DFL, LNT, and LNnT, ie, comprises, consists essentially of, or consists of 2'-FL, DFL, LNT, and LNnT.

[0063] In one embodiment, the HMO is a combination of 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL), i.e., it comprises, consists essentially of, or consists of 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL).

[0064] In one embodiment, the HMO is a combination of 2'-FL, DFL, LNT, LNnT, 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL), i.e., comprises, consists essentially of, or consists of 2'-FL, DFL, LNT, LNnT, 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL).

[0065] HMOs can be isolated or concentrated by well-known methods from milk secreted by mammals, such as, but not limited to, human, bovine, ovine, porcine, or caprine species. HMOs can also be produced by well-known methods using microbial fermentation, enzymatic processing, chemical synthesis, or a combination of these techniques. By way of example, using chemistry, LNnT can be prepared as described in WO2011 / 100980 and WO2013 / 044928, LNT can be synthesized as described in WO2012 / 155916 and WO2013 / 044928, mixtures of LNT and LNnT can be prepared as described in WO2013 / 091660, 2'-FL can be prepared as described in WO2010 / 115934 and WO2010 / 115935, 3-FL can be prepared as described in WO2013 / 139344, 6'-SL and salts thereof can be prepared as described in WO2010 / 100979, sialylated oligosaccharides can be prepared as described in WO2012 / 113404, and mixtures of human milk oligosaccharides can be prepared as described in WO2012 / 113405. As examples of enzymatic production, sialylated oligosaccharides can be produced as described in WO2012 / 007588, fucosylated oligosaccharides can be produced as described in WO2012 / 127410 and advantageously a diverse mixture of human milk oligosaccharides can be produced as described in WO2012 / 156897 and WO2012 / 156898. Furthermore, WO01 / 04341 and WO2007 / 101862 describe methods for producing core human milk oligosaccharides, optionally substituted by fucose or sialic acid, using genetically modified E. coli.

[0066] In any aspect of the present invention, HMO can be used as it is (neat) without carrier and / or diluent.In another embodiment, HMO is used in synthetic composition, which contains one or more inert carrier / diluent acceptable in nutritional or pharmaceutical composition, such as solvent (e.g. water, water / ethanol, oil, water / oil), dispersant, coating agent, absorption enhancer, controlled release agent, inert excipient (e.g. starch, polyol, granulating agent, microcrystalline cellulose, diluent, lubricant, binder, and disintegrant).These compositions do not contain prebiotics and / or probiotics.In another embodiment, HMO is used in synthetic pharmaceutical or nutritional composition, which may not contain prebiotics and / or probiotics.

[0067] The non-infant may be a healthy non-infant not suffering from sub-optimal executive function.

[0068] Executive functions can be measured by methods well known to those skilled in the art, for example, by assessing different cognitive abilities known to constitute executive functions, such as inhibition, working memory, cognitive flexibility, pattern recognition tests, etc., possibly comprehensively. For children, such methods are, for example, the Dimensional Change Card Sort (DCCS), and for adults, the Wisconsin Card Sorting Task. Participants are presented with a target card and asked to sort a series of bivalent test cards according to one dimension (e.g., color). In a post-switch phase, they are instructed to sort the same type of test cards according to the other dimension (e.g., shape).

[0069] A non-infant who is not suffering from suboptimal executive function will have a test score within the range considered to be normal (non-pathological), depending, for example, on the type and age of the non-infant.

[0070] As will be apparent to one of skill in the art, the HMOs or compositions comprising HMOs disclosed herein for use in improving executive function may also be used to prevent and / or treat suboptimal executive function in non-infant infants.

[0071] Thus, in another aspect of the invention, there is provided herein an HMO or a composition comprising an HMO for use in the prevention and / or treatment of suboptimal executive function.

[0072] In some embodiments, the non-infant can be a non-infant suffering from non-optimal executive function and therefore in need of enhanced executive function.

[0073] A human suffering from non-optimal executive function may be, for example, a non-infant who does not have a test score within the range considered normal (non-pathological) (on standard tests used to assess executive function), depending on the type and age of the non-infant. One skilled in the art will be able to determine when a non-infant suffers from non-optimal executive function.

[0074] Suboptimal executive function may be associated with various cognitive disorders, such as Attention Deficit Hyperactivity Disorder (ADHD), Alzheimer's disease, and vascular dementia. It is also thought to be more prevalent in non-infant infants who were born preterm or small for gestational age (SGA), and it is known that executive function may decline with age. Therefore, there may be a particular need to treat and / or prevent suboptimal executive function in these patient groups.

[0075] Thus, in a more specific embodiment, the non-infant in need of improved executive function is a non-infant, older adult or child who was born prematurely or small for gestational age (SGA) suffering from ADHD, Alzheimer's disease, or vascular dementia.

[0076] In another aspect of the invention, there is provided a method for preventing and / or treating non-optimal executive function in a non-infant comprising administering to said non-infant an HMO or a composition comprising an HMO as disclosed herein. The method may also optionally comprise identifying the non-infant suffering from non-optimal executive function.

[0077] In another aspect of the present invention, there is provided a method for improving executive function in a non-infant comprising the step of administering to said non-infant an HMO or a composition comprising an HMO as disclosed herein, and which may optionally include the step of identifying the non-infant suffering from suboptimal executive function.

[0078] In any aspect of the invention described above, the composition may be a pharmaceutical composition. The pharmaceutical composition may include pharmaceutically acceptable carriers, such as phosphate buffered saline, mixtures of ethanol in water, water, and emulsions such as oil / water, or water / oil emulsions, as well as various wetting agents, or excipients. The pharmaceutical composition may also include other substances that do not produce adverse, allergic, or other undesirable reactions when administered to non-infants. Carriers and other substances include solvents, dispersants, coating agents, absorption enhancers, controlled release agents, and one or more inert excipients such as starch, polyols, granulating agents, microcrystalline cellulose, diluents, lubricants, binders, and disintegrants. If necessary, tablet dosages of the composition may be coated by standard aqueous or nonaqueous techniques.

[0079] Pharmaceutical compositions can be orally administered, for example, as tablets, capsules, or pellets containing a predetermined amount, or as powders or granules containing a predetermined concentration, or as gels, pastes, solutions, suspensions, emulsions, syrups, boluses, electuaries, or slurries in aqueous or non-aqueous liquids containing a predetermined concentration. Orally administered compositions can include binders, lubricants, inert diluents, flavoring agents, and wetting agents. Orally administered compositions, such as tablets, can be appropriately coated and formulated to provide sustained, delayed, or controlled release of the mixture therein.

[0080] The pharmaceutical compositions may also be administered by rectal suppository, aerosol tube, nasogastric tube, or direct injection into the digestive tract.

[0081] The pharmaceutical composition may also include therapeutic agents such as antivirals, antibiotics, probiotics, analgesics, and anti-inflammatory agents. The appropriate dosage of these compositions for non-infant humans can be determined by conventional methods based on factors such as immune status, weight, and age. In some cases, the dosage will be at a concentration similar to that of HMOs present in human breast milk. The required dosage will generally range from about 1 g to about 20 g per day, and in some embodiments, from about 2 to about 15 g per day, for example, from about 3 g to about 7.5 g per day. The appropriate dosage regimen can be determined by conventional methods.

[0082] In any aspect of the present invention described above, the synthetic composition can also be a nutritional composition. It may contain protein, lipid and / or digestible carbohydrate sources and can be in powder or liquid form. The composition can be designed to be the sole source of nutrition or a dietary supplement.

[0083] Suitable protein sources include milk protein, soy protein, rice protein, pea protein, and oat protein, or mixtures thereof. Milk protein can be in the form of milk protein concentrate, milk protein isolate, whey protein, or casein, or mixtures thereof. Protein can be whole protein or hydrolyzed protein, either partially hydrolyzed or extensively hydrolyzed. Hydrolyzed protein offers the advantage of easy digestion, which can be important for non-infants with inflamed digestive tracts. Protein can also be provided in the form of free amino acids. Protein can comprise about 5% to about 30%, usually about 10% to 20%, of the energy of the nutritional composition.

[0084] The protein source may be a source of glutamine, threonine, cysteine, serine, proline, or combinations of these amino acids. The glutamine source may be glutamine dipeptide and / or glutamine-enriched protein. Glutamine may be included because glutamine is used by enterocytes as an energy source. Threonine, serine, and proline are important amino acids for the production of mucin, which lines the digestive tract and may improve mucosal healing. Cysteine ​​is the primary precursor of glutathione, which is important for the body's antioxidant defenses.

[0085] Suitable digestible carbohydrates include maltodextrin, hydrolyzed or modified starch or corn starch, glucose polymers, corn syrup, corn syrup solids, isomerized sugar, carbohydrates from rice, carbohydrates from peas, carbohydrates from potatoes, tapioca, sucrose, glucose, fructose, sucrose, lactose, honey, sugar alcohols (e.g., maltitol, erythritol, sorbitol), or mixtures thereof. Preferably, the composition does not contain lactose. In general, digestible carbohydrates provide about 35% to about 55% of the energy of the nutritional composition. Preferably, the nutritional composition does not contain lactose. A particularly suitable digestible carbohydrate is low dextrol equivalent (DE) maltodextrin.

[0086] Suitable carbohydrates include medium chain triglycerides (MCT) and long chain triglycerides (LCT). Preferably, the lipid is a mixture of MCT and LCT. For example, MCT may comprise about 30% to about 70% by weight of lipid, more specifically about 50% to about 60% by weight. MCT offers the advantage of being easier to digest, which may be important for non-infants with inflamed digestive tracts. Generally, lipid provides about 35% to about 50% of the energy of the nutritional composition. Lipid may include essential fatty acids (omega-3 and omega-6 fatty acids). Preferably, these polyunsaturated fatty acids provide less than about 30% of the total energy of the lipid source. It is particularly preferred that the composition includes an omega-3 polyunsaturated fatty acid source; for example, fish oil, docosahexaenoic acid, alpha-linolenic acid, and / or eicosapentaenoic acid. Polyunsaturated fatty acid may also be dihomo-gamma-linolenic acid (DGLA).

[0087] Suitable sources of long-chain triglycerides are rapeseed oil, sunflower seed oil, palm oil, soybean oil, milk fat, corn oil, high oleic oil, and soy lecithin. Fractionated coconut oil is a suitable source of medium-chain triglycerides. The lipid profile of the nutritional composition is preferably designed to have a ratio of polyunsaturated fatty acids omega-6 (n-6) to omega-3 (n-3) of about 4:1 to about 10:1. For example, the ratio of n-6 to n-3 fatty acids can be about 6:1 to about 9:1.

[0088] Preferably, the nutritional composition also contains vitamins and minerals. If the nutritional composition is intended as a sole source of nutrition, it preferably contains a complete vitamin and mineral profile. Examples of vitamins include vitamin A, B complex (such as B1, B2, B6, and B12), C, D, E, and K, niacin, and acidic vitamins such as pantothenic acid, folic acid, and biotin. Examples of minerals include calcium, iron, zinc, magnesium, iodine, copper, phosphorus, manganese, potassium, chromium, molybdenum, selenium, nickel, tin, silicon, vanadium, and boron. Vitamin D sources are particularly preferred.

[0089] The nutritional composition may also contain carotenoids such as lutein, lycopene, zeaxanthin, and beta-carotene. The total amount of carotenoids contained may vary from about 0.001 μg / ml to about 10 μg / ml. Lutein may be contained in an amount of about 0.001 μg / ml to about 10 μg / ml, preferably about 0.044 μg / ml to about 5 g / ml of lutein. Lycopene may be contained in an amount of about 0.001 μg / ml to about 10 μg / ml, preferably about 0.0185 mg / ml to about 5 g / ml of lycopene. Beta-carotene may be contained in an amount of about 0.001 μg / ml to about 10 mg / ml, for example about 0.034 μg / ml to about 5 μg / ml of beta-carotene.

[0090] The nutritional composition also preferably contains a reduced sodium concentration; for example, from about 300 mg / l to about 400 mg / l. The remaining electrolytes can be present at concentrations designed to meet the needs without placing an excessive renal solute load on renal function. For example, potassium is preferably present in the range of about 1180 to about 1300 mg / l; chloride is preferably present in the range of about 680 to about 800 mg / l.

[0091] The nutritional composition may also contain preservatives, emulsifiers, thickeners, buffers, fiber, and prebiotics (e.g., fructooligosaccharides, galactooligosaccharides), probiotics (e.g., B. animalis subsp. lactis BB-12, B. lactis HN019, B. lactis Bi07, B. infantis ATCC 15697, L. rhamnosus GG, L. rhamnosus HNOOl, L. acidophilus LA-5, L. acidophilus NCFM, L. fermentum CECT5716, B. longum BB536, B. longum AH1205, B. longum AH1206, B. breve M-16V, L. reuteri ATCC 55730, L. reuteri ATCC PTA-6485, L. reuteri DSM 17938), antioxidants / anti-inflammatory compounds such as tocopherols, carotenoids, ascorbic acid / vitamin C, ascorbyl palmitate, polyphenols, glutathione, and superoxide dismutase (melon), other biologically active factors (e.g., growth hormone, cytokines, TGF-β), colorants, flavorings, and other conventional ingredients such as stabilizers, lubricants, etc.

[0092] The nutritional composition may be in the form of a soluble powder, liquid concentrate, or ready-to-use formulation. The composition may be fed to non-infants via a nasogastric tube or orally. Various flavoring agents, fiber, and other additives may be present.

[0093] The nutritional composition can be produced by any manufacturing technique commonly used to prepare nutritional compositions in solid or liquid form. For example, the composition can be produced by combining various feeding solutions. A protein-in-lipid feeding solution can be produced by heating and mixing a lipid source, then adding an emulsifier (e.g., lecithin), fat-soluble vitamins, and at least a portion of the protein source with heating and stirring. A carbohydrate feeding solution is then produced by adding minerals, trace and ultratrace minerals, thickeners or suspending agents to water with heating and stirring. The resulting solution is held for 10 minutes with continued heating and stirring, after which carbohydrates (e.g., HMOs, and a digestible carbohydrate source) are added. The resulting feeding solutions are then mixed together with heating and stirring, the pH is adjusted to 6.6-7.0, and the composition is then subjected to a short period of high temperature treatment during which the composition is high temperature treated, emulsified, and homogenized, and then cooled. Water-soluble vitamins and ascorbic acid are added, the pH is adjusted to the desired range if necessary, flavorings are added, and water is added to reach the desired total solids.

[0094] For liquid products, the resulting solution can then be aseptically filled to produce a sterile packaged nutritional composition. In this form, the nutritional composition can be in the form of a ready-to-use liquid or a concentrated liquid. Alternatively, the composition can be spray dried and processed and packaged as a reconstitutable powder.

[0095] When the nutritional product is a ready-to-use nutritional liquid, the total concentration of HMOs in the liquid is about 0.0001% to about 2.0%, such as about 0.001% to about 1.5%, for example about 0.01% to about 1.0%, by weight of the liquid.When the nutritional product is a concentrated nutritional liquid, the total concentration of HMOs in the liquid is about 0.0002% to about 4.0%, such as about 0.002% to about 3.0%, for example about 0.02% to about 2.0%, by weight of the liquid.

[0096] The compound composition, preferably the nutritional composition, may also be in unit dosage form, such as a capsule, tablet, or sachet / stick package. For example, the compound composition, preferably the nutritional composition, may be in tablet or powder form, containing the HMO and one or more additional ingredients that aid in formulation and administration, such as diluents, excipients, antioxidants, lubricants, colorants, binders, disintegrants, etc.

[0097] Suitable diluents, excipients, lubricants, colorants, binders, and disintegrants include polyethylene, polyvinyl chloride, ethyl cellulose, polyarylate polymers and copolymers thereof, hydroxyethyl-cellulose, hydroxypropylmethyl-cellulose (HPMC), sodium carboxymethylcellulose, polyhydroxyethyl methacrylate (PHEMA), polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polyethylene oxide (PEO), or polyacrylamide (PA), carrageenan, sodium alginate, polycarbophil, polyacrylic acid, tragacanth, methylcellulose, pectin, natural gums, xanthan gum, guar gum, karaya gum, hypromellose, magnesium stearate, microcrystalline cellulose, and colloidal silicon dioxide.Suitable antioxidants are vitamin A, carotenoids, vitamin C, vitamin E, selenium, flavonoids, polyphenols, lycopene, lutein, lignans, coenzyme Q10 ("CoQIO"), and glutathione.

[0098] The unit dosage form may also contain various nutrients, such as macronutrients, particularly in the form of sachets or stick packages. Particularly preferred nutrients are the aforementioned vitamin D, omega-3 polyunsaturated fatty acids, and sources of probiotics.

[0099] The appropriate dosage of HMO or synthetic composition can be determined in a conventional manner based on factors such as the severity of symptoms, immune status, body weight and age.In some cases, the dosage will be at a concentration similar to that present in HMO present in human breast milk.The required amount will generally be in the range of about 1g to about 20g per day, in some embodiments, about 1g to about 15g per day, for example, about 2g to about 10g per day, in some embodiments, about 3g to about 7.5g per day.The appropriate dosage regimen can be determined by conventional methods.Ideally, a dosage of about 2g to about 10g per day is administered, with the exact amount being selected depending on whether symptoms are being treated (generally high dosage) or whether secondary prevention is intended (low dosage).

[0100] The HMO or synthetic composition may be presented in the form of a pack containing at least 14 individual daily doses of effective amounts of human milk oligosaccharides. The daily doses are preferably in the form of sachets / stick packaging, but may be in any suitable form. Each dose preferably contains about 1 g to about 20 g, more preferably about 2 g to about 15 g, for example about 2 g to about 10 g of human milk oligosaccharides. Preferably, the pack contains at least 21 days' doses, more preferably at least 28 days' doses. The most suitable packs contain a sufficient amount for 4 weeks or one month. The pack may include instructions for use.

[0101] Appropriate dosage can be determined based on several factors, such as body weight and / or disease state, severity of symptoms, incidence and / or severity of side effects, and administration method.Effective amount can be any amount that improves executive function in non-infants.Improvement of executive function can be measured by well-known tests, such as those detailed above.Improvement of executive function may only be detectable after 6 months, 1 year, for example, after 5 years, after 10 years, after 20 years.Based on the information in this specification and knowledge in the art, it is well within the scope of a person skilled in the art to determine effective amount.

[0102] The appropriate dose range can be determined by methods known to those skilled in the art.During the initial treatment phase, the dose can be higher (e.g., 3g to 15g per day, preferably 4g to 10g per day, more preferably 4g to 7.5g per day).During the maintenance phase, the dose can be reduced (e.g., 1 to 10g per day, preferably 2mg to 5g per day, more preferably 2g to 5g per day). When the composition comprising HMO comprises 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL), it may be particularly beneficial when the 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL) are contained in the composition in a weight ratio of between 10:1 and 1:10, for example between 10:1 and 2:1, between 8:1 and 3:1, between 6:1 and 3:1, between 5:1 and 3:1, between 5:1 and 4:1, or between 4.7:1 and 4.1:1.

[0103] The duration of treatment can be determined based on several factors, such as, for example, body weight and / or disease state, severity of symptoms, incidence and / or severity of side effects, etc. Preferably, the duration of treatment is at least 14 days, more preferably at least 3 weeks; even more preferably 4 weeks or more. The synthetic composition or HMO can also be taken chronically.

[0104] Some aspects of the present invention relate to the management of dietary intake of non-infant humans required for improving executive function. - one or more human milk oligosaccharides (HMOs), - a synthetic composition comprising one or more human milk oligosaccharides (HMOs), or - A pack containing at least 14 individual daily doses of an effective amount of one or more human milk oligosaccharides, The HMO, the synthetic composition and the pack are disclosed above.

[0105] It should be understood that all features of the invention disclosed herein can be freely combined, and changes and modifications can be made without departing from the scope of the invention as defined in the claims. Moreover, where certain features have known equivalents, such equivalents are incorporated as if specifically referred to herein.

[0106] As used in this disclosure and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly indicates otherwise. So, for example, reference to "an ingredient" or "the ingredient" includes two or more ingredients. The term "and / or" used in the context of "X and / or Y" should be interpreted as "X," "Y," or "X and Y." As used herein, the term "for example," especially when followed by a list of terms, is merely exemplary and explanatory and should not be considered exclusive or inclusive.

[0107] As used herein, "about" should be understood to refer to a number within a range of values, for example, within -10% to +10% of the referenced number, preferably within -5% to +5% of the referenced number, more preferably within -1% to +1% of the referenced number, and most preferably within -0.1% to +0.1% of the referenced number. A range "between" two values ​​includes those two values. Furthermore, all numerical ranges herein should be understood to include all integers, wholes, or fractions within the range. Furthermore, these numerical ranges should be understood to provide support for claims regarding any number or subset of numbers within the range. For example, a disclosure of 1 to 10 should be understood to support ranges such as 1 to 8, 3 to 7, 1 to 9, 3.6 to 4.6, 3.5 to 9.9, etc.

[0108] All percentages expressed herein are by weight of the total weight of the composition unless otherwise stated. When pH is mentioned, the value corresponds to the pH measured at 25°C by standard equipment.

[0109] The relative terms "improvement" and "reduction" refer to the effect of the HMO or composition comprising HMO disclosed herein on executive function (e.g., effect on different cognitive abilities known to constitute executive function, such as attention or impulsivity, working memory, cognitive flexibility). This would also need to be considered overall in non-infants in comparison with non-infants not administered HMO or composition comprising HMO. It is well within the scope of the skilled artisan to evaluate improvement, increase, or enhancement. The improvement of executive function (different cognitive abilities known to constitute executive function, such as cognitive flexibility, working memory, attention and / or reduction of impulsivity) may only be detectable after 1 year, for example, after 5 years, after 10 years, after 20 years.

[0110] The compositions disclosed herein may lack any element not specifically disclosed herein. Thus, the disclosure of an embodiment using the term "comprising" includes the disclosure of an embodiment "consisting essentially of" and "consisting of" the specified components. Similarly, the methods disclosed herein may lack any step not specifically disclosed herein. Thus, the disclosure of an embodiment using the term "comprising" includes the disclosure of an embodiment "consisting essentially of" and "consisting of" the specified steps. Furthermore, the description of some steps as "optional" does not imply that other steps not expressly described as optional are necessarily required. If the sentence simply refers to a composition, it may be a nutritional composition.

[0111] Below is a series of non-limiting examples that serve to illustrate the invention. EXAMPLES

[0112] Example 1 Methods - Behavioral research Fifty-three female piglets (Gottingen minipigs, Ellegaard, Denmark) were subjected to the hole-board test after weaning. At 1 week of age, the formula-fed (N=45) piglets were separated from the sows and mixed groups of two piglets were housed in pens (2.5mx1m) with increased shredded straw as bedding, squeeze bowls and dog beds. In addition, eight piglets were mated and kept with three sows until 10 weeks of age (naturally fed piglets). After weaning from mother's milk, at 10 weeks of age, the naturally fed piglets were kept in similar conditions to the formula fed piglets. Two metal chains were added to the pens as further enrichment material.

[0113] Treatment.Milk interventions ranged from 1 to 10 weeks of age. At 1 week of age, formula-fed piglets were randomly assigned to receive one of four mixed milks enriched with different prebiotic mixtures (Milk A: 3'-SL and 6'-SL, Milk B: 2'-FL+DFL+LNT+LNnT, Milk C: 2'-FL+DFL+LNT+LNnT+3'-SL+6'-SL) or without prebiotics (control milk). Naturally-fed piglets were breast-fed for the entire duration of the nutritional intervention. After weaning, 10-week-old piglets were fed a high-energy obesogenic diet. This experimental setup resulted in five treatment groups: Milk A (N=12), Milk B (N=12), Milk C (N=10), Control Milk (N=11), and Naturally-fed (N=8).

[0114] Both pigs in each pen were tested individually in a spatial hole-board task to assess the performance of spatial cognition (memory and learning). The hole-board arena (3mx3m) had black wooden 80cm high walls and four entrances with guillotine doors. In the arena, 16 grey metal buckets (φ12cm-H12cm) were embedded in the floor in a 4x4 matrix, four of which contained apple pieces (~12x12x20mm). The piglets in the arena could see the walls and ceiling of the room by means of ventilation ducts and light tubes. The piglets were food deprived overnight for the entire duration of the hole-board test.

[0115] From approximately 16.5 to 19 weeks of age, each piglet was subjected to two group trials per day (conducted several minutes apart) for 12 consecutive days, i.e., 24 acquisition trials. A different entrance was used each day, with two different entrances per test day (i.e., one entrance per test). A test started when the piglet placed all four paws on the hole-board arena and ended when the piglet found all four rewards or after 180 s. A clicking sound was produced each time the piglet went to the food bucket for the first time to promote learning. When the piglet completed the task (i.e., found all four rewards within 180 s), the exit (south) guillotine door was opened and the piglet received half a white grape. If the piglet did not complete the task within 180 s, a police car siren sound was produced; the piglet did not receive any reward. After two trials with two piglets per pen, animals from all pens were returned to their original home pens and given their morning milk ration.

[0116] After completing the acquisition phase, each piglet was subjected to 16 reversal trials, two group trials per day for eight consecutive days. The procedure was similar to that of the acquisition phase, except that piglets were assigned different configurations of food buckets.

[0117] The following parameters were scored live using The Observer XT 10 (Noldus Information Technology, Wageningen, The Netherlands): all visits and revisits to all buckets during the test, latency to visit all buckets, trial duration, total number of defecation, urination, and escape attempts. From the parameters recorded during the test, variables were calculated post hoc according to van der Staay et al. Neurosci. Biobehav. Rev. 36, 379 (2012) (Table 1). [Table 1]

[0118] During the acquisition phase, all pigs showed an increase in executive function performance, which could be visualized by WM, RM, and total WM scores that increased linearly over time, and by the number of WM, RM, and total WM errors that decreased linearly over time.

[0119] During the reversal phase, all piglets showed an increase in executive function, visualized by WM, RM, and total WM scores that increased linearly with time, and by the number of WM, RM, and total WM errors that decreased linearly with time. This improvement in RM, WM, and total WM performance was reduced in the control milk group compared to naturally fed piglets (significantly for total WM and RM), and surprisingly, these deficits were restored in the presence of HMO in the milk to levels similar to those observed in the naturally fed group (see Figures 1-3).

[0120] Example 2 Animals and housing conditions Adult wild-type (WT) B6.129 and heterozygous (HZ) B6.129-St6gal1 tm2JxmBreeding pairs (four males and four females, and three males and four females, respectively) were purchased from a commercial breeder (The Jackson Laboratory). On arrival, same-sex mice were housed in same-sex groups of two to three in type 1 polycarbonate cages (33.0x13.0x14.0cm) with sawdust bedding, enrichment bags (Mucedola, Settimo Milanese, Italy), metal surfaces and ad libitum water and food pellets. Mice were kept in an air-conditioned room (temperature 21 ± 1°C and relative humidity 60 ± 10%) on a 12-h reversed light-dark cycle (lights on at 7:00 PM). Two weeks after arrival, breeding pairs of three (one male, two females) were formed. After 2 weeks of mating, male mice were removed and females were housed individually in standard type 1 cages. Females were checked daily for birth and the day of birth was designated postnatal day (PND) 0. Dams and pups were left alone until weaning (PND25), except for weekly cage cleaning. At weaning, male and female mice were separated and housed in same-sex littermates; male mice were further marked with ear clips, and ear tissue removed by this procedure was used for genotyping. Homozygous knockout (KO) and WT mice were then used for experiments.

[0121] Breeding procedures and husbandry Fourteen wild-type (WT) and fourteen St6Gal1 homozygous (hereafter KO) female mice were mated with seven WT and seven KO male mice, respectively. Of this batch, ten WT and ten KO dams gave birth to viable pups. The day of birth was designated postnatal day (PND) 0. The rearing procedure, which took place between 10:00 and 13:00 (see Fig. 1 for details), required the use of four dams (two WT and two KO) simultaneously. Therefore, to minimize the number of subjects abandoned due to the absence of a rearing dam, the rearing procedure was performed between 24 and 60 hours after birth. On the rearing day, we first removed the dams from their cages, then sexed them and marked the pups by puncturing their toes with tattoo ink. After the sexing and marking procedures were completed, the pups were transferred to the cage with the rearing dam and covered with sawdust. Each pup was given to a breeding dam to ensure that all experimental subjects were placed under the same conditions. Each dam raised mixed litters consisting of WT and KO male and female pups (1:1 ratios for all variables whenever possible). At eye opening and adulthood, prefrontal cortex and hippocampal brain samples were collected for gene expression analysis.

[0122] The following experimental groups were constructed: -WT offspring bred to WT dams (WT vs WT) -WT offspring bred to KO dams (WT vs. KO) -KO offspring bred to WT dams (KO vs. WT) -KO offspring bred by KO dams (KO vs KO)

[0123] Total RNA extraction and QC Total RNA was extracted using the Agencourt RNAdvance Tissue Kit (Beckman Coulter): lysis was performed in 450 μl. 400 μl of lysate was extracted. Elution volume 50 μl. Quantification is performed using the Quant It Ribogreen assay (Life Technologies). QC assessment is performed using the Standard sensitivity RNA kit (Agilent) on a Fragment Analyzer 96.

[0124] Sample library preparation Libraries are generated using Lexogen's QuantSeq 3' mRNA-Seq Library Prep Kit (FWD) HT for Illumina, which is designed to generate Illumina-compatible sequences close to the 3' end of polyadenylated RNA.

[0125] As the kit uses total RNA as input, no prior poly(A) enrichment or rRNA depletion is required. Library generation is initiated by oligo-dT priming, which contains the Illumina-specific Read2 linker sequence. After first strand synthesis, the RNA is removed. Second strand synthesis is initiated by random priming and DNA polymerase. The random primer contains the Illumina-specific Read1 linker sequence. No purification is required between first and second strand synthesis. Second strand synthesis is followed by a magnetic bead purification step. Sequences required for cluster formation for sequencing are introduced during the library amplification step. Double-stranded cDNA is amplified by PCR. In this step, individual barcode indexes are introduced and samples are multiplexed. NGS reads are directed towards the poly(A) tail and correspond directly to the mRNA. See Figure 1 for details. Libraries were quantified by Quant it Picogreen (Life Technologies). Size patterns are controlled using the High Sensitivity NGS Fragment Analysis kit (Agilent) in a Fragment Analyzer. Libraries are pooled in equimolar ratios (i.e. equal amounts of each sample library) and clustered at a concentration of 9 pM on a single read (SR) sequencing flow cell (Illumina). Sequencing is performed for 65 cycles on a HiSeq 2500 (Illumina) using a HiSeq SR Cluster Kit v4 cBot, HiSeq SBS Kit V4 50 cycle kit (Sequencing by Synthesis). Primary data quality control is performed during the sequencing run to ensure optimal flow cell loading (cluster density) and to confirm the quality metrics of the sequencing run (QC30). The optimal density numbers of clusters detected by image analysis are 850 and 1000K / mm 2Running at an optimal cluster density involves finding a balance between under-clustering, which has high quality data but low data output, and over-clustering, which may result in poor running performance. > The Q30 percentage means the percentage of bases with a quality Phred score of 30 or higher. It is a measure of the quality of the generated nucleobase identifications. The Phred score is logarithmically related to the error probability. A Phred score equal to 30 means a probability of 1 in 1000 of incorrect base assignment, therefore the accuracy of the base readout is 99.9%. Regarding Illumina specifications, the score must be at least 80%.

[0126] 4. Data Analysis Genes with very low counts are unlikely to be differentially expressed between groups. Low-expressing genes were filtered by selecting only genes with at least 5 reads in at least 8 samples. The threshold number of samples ensures that genes are expressed in the smallest group (containing 8 samples). The filtering procedure is performed at a CPM value that takes into account the library size. In this case, the CPM value corresponds to a threshold of 2.427. Also, unannotated genes were discarded. These filtering criteria retained 11523 features. Normalization was performed using the TMM method.

[0127] result The expression of multiple genes related to myelination was reduced in mice fed 6'-SL-free milk only in the prefrontal cortex during childhood, but not in the hippocampus during adolescence or at either of the two stages. Figure 4 shows that the expression of two key proteins of myelin, myelin basic protein (MBP) and myelin-associated glycoprotein (MAG), was reduced by 50% in mice fed 6'-SL-free milk (gene expression was quantified in fold change compared to control mice). The impact of the absence of 6'-SL during childhood was investigated in the Kyoto Encyclopedia of Genes and Genomes pathways, identifying that both myelination and myelination pathways were downregulated in mice fed 6'-SL-free milk. This was also visualized in the prefrontal cortex and not in the hippocampus (either during adolescence or childhood). These results clearly indicate that the presence of 6'-SL in breast milk is necessary to observe optimal myelination of the prefrontal cortex, especially during childhood. Interestingly, the prefrontal cortex is known to be a key region of the brain mediating executive functions.

[0128] Example 3 - Nutritional Composition The ready-to-serve nutritional composition is made from water, maltodextrin, corn syrup, sugar, milk protein concentrate, vegetable oils (canola, high oleic sunflower, and corn), soy protein isolate, gum acacia, flavors, 2'-FL and DFL, potassium citrate, magnesium phosphate, cellulose gel and gums, calcium carbonate, sodium ascorbate, soy lecithin, choline bitartrate, calcium phosphate, alpha-tocopheryl acetate, ascorbic acid, carrageenan gum, ferric pyrophosphate, flavors, sweetener (stevia), vitamin A palmitate, niacinamide, vitamin D3, calcium pantothenate, manganese sulfate, copper sulfate, pyridoxine hydrochloride, thiamine hydrochloride, beta carotene, riboflavin, chromium chloride, folic acid, biotin, potassium iodide, phytonadione, sodium selenite, sodium molybdate, and vitamin B12.

[0129] The composition provides a dietary supplement that contains a good source of protein, low fat, vitamins, minerals, and antioxidants, and meets the FODMAP criteria. Additionally, the composition contains 2'-FL and DFL, which can promote the growth of beneficial gut bacteria and improve intestinal barrier function.

[0130] Example 4 - Capsule Composition Approximately 1 g of 2'-FL and DFL are filled into 000 gelatin capsules using a filling machine to produce the capsules. The capsules are then closed. The 2'-FL and DFL are in the form of a free-flowing powder.

Claims

1. In non-infants, - A synthetic composition for use in improving executive function by improving, increasing or enhancing working memory, comprising at least one human milk oligosaccharide (HMO), said human milk oligosaccharide comprising a combination of 2'-fucosyllactose (2'-FL), 3,2'-difucosyllactose (DFL), lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT) or a combination of 2'-FL, DFL, LNT and LNnT, 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL).

2. In non-infants, - a pack for use in improving executive functions by improving, increasing or enhancing working memory, comprising at least 14 individual daily doses of an effective amount of at least one human milk oligosaccharide (HMO), said human milk oligosaccharide comprising a combination of 2'-fucosyllactose (2'-FL), 3,2'-difucosyllactose (DFL), lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT) or a combination of 2'-FL, DFL, LNT and LNnT, 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL).

3. The human milk oligosaccharides are -2'-FL, DFL, LNT, and LNnT, or -2'-FL, DFL, LNT, LNnT, 3'-SL and 6'-SL 3. A synthetic composition or pack according to claim 1 or 2, which consists or essentially consists of:

4. The non-infant, - have Attention Deficit Hyperactivity Disorder (ADHD), Alzheimer's disease, depression, anxiety, bipolar disorder, schizophrenia, obsessive-compulsive disorder (OCD), autism, Alzheimer's disease, frontotemporal dementia, dementia with Lewy bodies, Tourette's syndrome, post-traumatic stress disorder (PTSD), traumatic brain injury, or vascular dementia, - an elderly person, or - is or has been a preterm or small for gestational age baby, A synthetic composition or pack according to any one of claims 1 to 3.

5. A composition or pack according to any one of claims 1 to 4, wherein said composition or said daily dose is administered in combination with cognitive behavioral therapy.

6. A composition or pack according to any one of claims 1 to 5, wherein said composition or said daily dose comprises HMO in an amount between 0.5g and 15g.

7. A composition or pack according to any one of claims 1 to 6, wherein said composition or said daily dose is administered to said non-infant for a period of at least one week.

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