INFANT FORMULA WITH MILK FAT FOR PROMOTING HEALTHY GROWTH

Abstract

The present invention relates to formulae for infants comprising milk fat for rendering the growth trajectory or body development during the first year of life more similar to that observed for human milk fed infants.

Claims

1.-23. (canceled)

24. A method of promoting postnatal growth trajectory or body development in an infant towards a growth trajectory or body development which is similar to the growth trajectory or body development observed in human milk fed infants, the method comprising administering to the infant an infant formula and/or a follow on formula comprising: (a) 1.25 to 5 g protein/100 kcal: (b) 6 to 18 g digestible carbohydrate/100 kcal; and (c) 3 to 7 g lipid/100 kcal, wherein the lipid comprises (i) 30 to 90 wt. % vegetable fat based on total lipid, and (ii) 10 to 70 wt. %, based on total lipid, of mammalian milk lipid derived from the group consisting of butter, butter fat, butter oil, and anhydrous milk fat, and (iii) 5 to 35 wt. % poly-unsaturated fatty acids (PUFA), based on total fatty acids, comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in a weight ratio of 2 to 20.

25. The method according to claim 24, wherein the growth trajectory or body development is over the first 12 months of life of the infant.

26. The method according to claim 24, wherein the growth trajectory or body development is selected from the group consisting of the trajectory or development for body weight, weight for length and/or body mass index (BMI).

27. The method according to claim 24, wherein at 12 months the infant has a weight and/or BMI and/or weight for length that is approximate to the weight and/or BMI and/or weight for length at 12 months of human milk fed infants.

28. The method according to claim 24, wherein at 12 months the infant has a weight and/or BMI and/or weight for length that is approximate to the weight and/or BMI and/or weight for length at 12 months according to the WHO Child Growth Standards of human milk fed infants.

29. The method according to claim 24, wherein the growth trajectory or body development is the trajectory or development for head circumference.

30. The method according to claim 29, wherein at 12 months the infant has a head circumference that is approximate to the head circumference at 12 months of human milk fed infants.

31. The method according to claim 29, wherein at 12 months the infant has a head circumference that is approximate to the head circumference at 12 months according to the WHO Child Growth Standards of human milk fed infants

32. The method according to claim 24, wherein the growth trajectory or body development is the trajectory or development for skinfold thickness.

33. The method according to claim 32, wherein the skinfold thickness is subscapular skinfold thickness and/or triceps skinfold thickness.

34. The method according to claim 24, wherein at 12 months the infant has a skinfold thickness, at 12 months of human milk fed infants.

35. The method according to claim 34, wherein the skinfold thickness is subscapular skinfold thickness and/or triceps.

36. The method according to claim 24, wherein at 12 months the infant has a subscapular skinfold thickness and/or triceps skinfold thickness that is approximate to the subscapular skinfold thickness and/or triceps skinfold thickness at 12 months according to the WHO Child Growth Standards of human milk fed infants.

37. The method according to claim 24, wherein the infant is a term born infant.

38. The method according to claim 24, wherein the infant is a male infant.

39. The method according to claim 24, wherein the nutritional composition comprises lipid globules that have a mode diameter, based on volume of at least 1.0 ?m and/or a diameter of 2 to 12 ?m in an amount of at least 45 volume % based on total lipid.

40. The method according to claim 24, wherein the nutritional composition comprises lipid globules that have on the surface at least partly a layer of phospholipids.

41. The method according to claim 24, wherein the nutritional composition comprises lipid globules that have (i) a mode diameter, based on volume of at least 1.0 ?m and/or a diameter of 2 to 12 ?m in an amount of at least 45 volume % based on total lipid and (ii) on the surface at least partly a layer of phospholipids.

42. The method according to claim 24, which comprises at least 0.5 wt. % phospholipids based on total lipid.

43. The method according to claim 24, wherein the phospholipids comprise at least 15 wt. % sphingomyelin based on total phospholipids.

44. The method according to claim 24, wherein formula comprises phospholipids that are derived from or form part of the milk fat globule membrane (MFGM).

45. The method according to claim 24, wherein the formula is a powder, suitable to reconstitute with water to a ready to drink formula.

46. A method of (i) promoting a balanced growth trajectory or body development in an infant and/or (ii) preventing or reducing the risk of an unbalanced growth trajectory or body development in an infant, the method comprising administering to the infant an infant formula and/or a follow on formula comprising: (a) 1.25 to 5 g protein/100 kcal: (b) 6 to 18 g digestible carbohydrate/100 kcal; and (c) 3 to 7 g lipid/100 kcal, wherein the lipid comprises (i) 30 to 90 wt. % vegetable fat based on total lipid, and (ii) 10 to 70 wt. %, based on total lipid, of mammalian milk lipid derived from the group consisting of butter, butter fat, butter oil, and anhydrous milk fat, and (iii) 5 to 35 wt. % poly-unsaturated fatty acids (PUFA), based on total fatty acids, comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in a weight ratio of 2 to 20.

47. A method for promoting postnatal growth trajectory or body development in an infant towards a growth trajectory or body development which is similar to the growth trajectory or body development observed in human milk fed infants, the method comprising administering to the infant an infant formula and/or follow on formula comprising: (a) 1.25 to 5 g protein/100 kcal; (b) 6 to 18 g digestible carbohydrate/100 kcal; and (c) 3 to 7 g lipid/100 kcal, wherein the lipid comprises (i) 5 to 35 wt. % poly-unsaturated fatty acids (PUFA), based on total fatty acids, comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in a weight ratio of 2 to 20, and (ii) triglycerides comprising at least 10 wt. % palmitic acid based on total fatty acids, and wherein at least 15% of the palmitic acid is present at the sn-2 position of the triglycerides.

Description

DESCRIPTION OF THE FIGURES

[0103] In the graphs in FIGS. 1-3, the x-as represents the number of weeks after birth in the life of an infant. The y-axis represents the z-score compared to the WHO Child Growth Standard of breastfed infants. Data points at 0, 17 weeks and 52 weeks are represented slightly shifted to one another so that the confidence intervals become visible.

[0104] FIG. 1 shows mean and 95% confidence interval (CI) of weight-for-age-z-score, by age category, ITT group.

[0105] FIG. 2 shows mean and 95% CI of BMI-for-age z-score, by age category, ITT group.

[0106] FIG. 3 shows mean and 95% CI of weight-for-length z-score, by age category, ITT group. In the examples data is collected at 17 weeks and at 52 weeks of age. This is interpreted as corresponding to 4 months and 12 months of age respectively. In other words, in the context of the present invention, 4 months of age is considered the same as 17 weeks of age and 12 months of age is considered the same as 52 weeks of age.

EXAMPLES

Example 1: Experimental and Control Formula

Diet 1: Standard Nutrilon 1

[0107] An infant formula with per 100 ml ready to drink formula 66 kcal, 1.3 g protein (whey protein and casein in a 6/4 w/w ratio), 7.3 g digestible carbohydrates (mainly lactose), 3.4 g fat and 0.8 g short chain galacto-oligosaccharides (source VivinalGOS) and long chain fructo-oligosaccharides (source RaftilinHP) in a 9/1 w/w ratio, and minerals, vitamins trace elements and other micronutrients as known in the art and in compliance with directives for infant formula. The formula is provided as a powder with the instruction to reconstitute with water. About 13.6 g powder was reconstituted to 100 ml ready to drink infant formula.

[0108] The fat component comprised mainly vegetable fat (blend of palm oil, low erucic acid rape seed oil, coconut oil, high oleic sunflower oil, sunflower oil, a small amount of soy lecithin (0.13 wt. %) and about 1.5 wt. % of an LC-PUFA premix (fish oil and microbial oil). The lipid globules had a mode diameter, based on volume, of about 0.5 ?m, and the volume % of lipid globules with a mode between 2 and 12 ?m was below 10.

Diet 2: Experimental Infant Formula

[0109] An infant formula similar as diet 1, except for the following:

[0110] The fat component consisted of about 51 wt. % vegetable fat (blend of low erucic acid rape seed oil, coconut oil, high oleic sunflower oil, sunflower oil), about 44 wt. % bovine anhydrous milk fat, 1.5 wt. % LC-PUFA containing oil (fish oil and microbial oil), 0.13 wt. % soy lecithin, about 3.6 wt. % milk fat derived from buttermilk rich in milk phospholipids or milk fat globule membranes (milk phospholipids are about 1.5 wt. % based on total lipid).

[0111] The lipid globules had a mode diameter, based on volume, of about 5.6 ?m, and the volume % of lipid globules with a mode between 2 and 12 ?m was above 45.

[0112] The fatty acid composition is very similar between diet 1 and 2, in saturated, mono unsaturated and poly unsaturated acids, and in n3 and n6 PUFA content. The amount of palmitic acid was 18.4 wt. % and 17.7 wt. % (based on total fatty acids) for diet 1 and 2, respectively. For diet 2 about 36 wt. % of the palmitic acid residues was in the sn2 position, for diet 1 this was about 13 wt. %. The amount of C4:0 (butyric acid) was 0.10 wt. % in diet 1 and 1.39 wt. % in diet 2, C6:0 (caproic acid) was 0.24 wt. % in diet 1 and 0.98 wt. % in diet 2. The wt. % are based on total lipid in the infant formula.

Example 2: Study Protocol and Study Population

[0113] After parent(s)/legal guardian(s) have signed informed consent, exclusively formula fed infants eligible for participation were randomised to receive either the experimental product or the control product for a double-blind period of maximally 17 weeks (depending on their age at study entry). Exclusively breastfed infants participated in the reference group and had the same visit schedule and study assessments as the randomised infants. At the first visit, baseline and birth data were collected, and the study product and diaries were provided to the parent(s). Further study visits were conducted at 5, 8, 13 and 17 weeks of age. Information and anthropometrical measurements were collected during the visits. During the visit at 52 weeks (an optional extension of the study), anthropometrical measurements were collected. 4 countries with 17 sites participated, and in total 313 subjects were enrolled; 6 sites in the Netherlands (121 subjects), 3 sites in France (13 subjects), 7 sites in Belgium (158 subjects), and 1 site in Singapore (21 subjects). Of the total of 313 enrolled subjects, 223 were randomised and 88 were included in the breastfed reference group, 2 subjects were screen failures and were consequently not randomised.

[0114] The All-Subjects-Treated (AST) data set consisted of all subjects randomised (ASR, n=223) who received at least some study product. Subjects (n=8) with sufficient evidence that no study product was consumed were considered as non-treated, and were not included in the AST group (n=215).

[0115] The ITT data set consisted of all subjects from the ASR group (ITT=ASR). Results from the ITT analysis reflect the effects on the targeted population in a real clinical situation/estimates the effect (effectiveness) of the treatment policy. Subjects' data were analysed as randomised.

[0116] The Per-Protocol (PP) analysis restricts the analysis to the subjects who fulfil the protocol in the terms of the eligibility, interventions, instructions/restrictions and outcome assessment. The PP data set consisted of all subjects and/or subjects' visits from the ITT data set without any major protocol deviations. Thus, the PP dataset was not limited to subjects who completed the study, and the number of subjects per visits varies. Results from the PP analysis estimate the effect (efficacy) of the treatment. Subjects' data were analysed as treated. The following rules have been applied for exclusion of subjects from the PP data set: Age at baseline (=visit 1)>35 days, birth weight missing or is <9.96th or >90.04th percentile (based on WHO Child growth standard references), Head circumference at inclusion is outside?2.04 SD percentile (based on WHO Child growth standard references), not having at least one valid post-baseline visit. Study product consumption started ?6 days after baseline, having received a different study product as his/her twin sibling, no study product was consumed, relevant medical history, i.e. illnesses/conditions as identified by the Medical Monitor. The following rules have been applied for exclusion of distinct visits from the PP data set: Any visit >3 days after stop of study product intake, regardless if stop was temporarily or not, any visit >3 days after start of other formula feeding, any visit >3 days after start of solid feeding. 49 randomised subjects plus certain visits were excluded.

[0117] For the non-randomised breastfed reference group data sets corresponding to the ITT and PP populations of the randomised infants have been defined, too. Correspondingly to the ITT data set, a full breast fed group (FBF) has been defined, no breast fed subjects were excluded. Correspondingly to the PP dataset, a Protocol Compliant Breastfed Reference (PCBF*) data set has been defined, applying the relevant rules as defined for the PP dataset. The following rules have been applied for exclusion of subjects from the PCBF data set: Age at baseline>35 days, birth weight missing or is <9.96th or >90.04th percentile (based on WHO Child growth standard references), Head circumference at inclusion is outside ?2.04 SD (based on WHO Child growth standard references), not having at least one valid post-baseline visit, or relevant medical history, i.e. illnesses/conditions as identified by the Medical Monitor. The following rules have been applied for exclusion of distinct visits from the PCBF data set: Any visit>3 days after stop of breastfeeding, in case stop of breastfeeding occurred before 13 weeks of age, any visit>3 days after start of other formula feeding, in case start of other formula feeding occurred before 13 weeks of age, any visit>3 days after start of solid feeding, in case start of other solid feeding occurred before visit 4. 11 breastfed subjects plus certain visits were excluded.

[0118] Subjects (either randomised or breastfed) who were included in PP/PCBF dataset up and including visit at 17 weeks of age and participated in the optional extension, were included in PP dataset at visit at 52 weeks.

[0119] There were no statistical significant differences between the intervention groups within different dataset (PP, ITT) on the stratification factors sex, age at baseline (<14 days/>14 days), regions (Europe vs. Asia). There was no difference in the duration in the study between the intervention groups.

[0120] The analysis of growth (weight and BMI at 52 weeks) was performed using parametric growth curves (PGC), correcting for the stratification factors described above. This approach assumes a parametric function of time (i.e. age of subject) and thus, describes the development of growth parameters (i.e. weight, BMI) over time by a second order polynomial function. It does not require the study subjects to be measured at the same set of time points. The resulting parameters are compared to assess differences between the curves. Sensitivity analyses to confirm model suitability were performed using General Linear Modelling (ANCOVA) and Arbitrary Mean Models, where time modelled as a categorical variable.

[0121] For the analysis of z-scores, each anthropometric measurement of subjects was normalized by using WHO Child growth standard z-score references which are age and gender dependent standardised values. A SAS macro (provided by WHO, http://www.who.int/childgrowth/software/en/) was used while normalizing the absolute anthropometric measurements of subjects, no correction for the stratification factors was performed. The analysis of z-scores was performed by age-category, selecting anthropometrical data collected within a window of ?10 days around a visit, taking the real age of the subjects into account. Age-categories are: Birth, 10?10 days at study entry, 35?10 days at week 5, 56?10 days at week 8, 91?10 days at week 13, 119?10 days at week 17, and 365?10 days at 52 weeks.

[0122] To conclude equivalence of weight gain from baseline visit (randomisation) until the age of 17 weeks in infants receiving the experimental product compared to infants receiving the control product, the two-sided 90% confidence intervals for the differences in mean weight gain should lie entirely between ?0.5 SD and +0.5 SD margins, with a minimum of 3 g/d and a maximum of 5 g/d. The equivalence analysis was performed using parametric growth curves, correcting for the stratification factors described above. This approach assumes a parametric function of time (i.e. age of subject) and thus, describes the development of growth parameters (i.e. weight) over time by a second order polynomial function. It does not require the study subjects to be measured at the same set of time points. The resulting parameters are compared to assess differences between the curves. Sensitivity analyses to confirm model suitability were performed using General Linear Modelling (ANCOVA) and Arbitrary Mean Models, where time modelled as a categorical variable.

Example 3: Results Growth Trajectories and Body Development

Weight

[0123] The median weight (kg) of boys and of girls (included in the three study groups) was shown to be well within the z-score of ?1 and +1 of the WHO Child growth standard z-scores, for the ITT and for the PP dataset (data not shown).

[0124] The weight gain per day in the experimental group was equivalent compared to the breastfed reference group (post hoc analysis) from visit at baseline until the age of 17 weeks, in the PP as well in the ITT datasets, using an equivalence margin of ?3 g/day. Equivalence of weight gain per day was demonstrated for the experimental formula compared to standard formula group (PP and ITT population), even when only selecting infants enrolled before 14 days of age.

[0125] The mean weight-for-age z-scores per visit and the corresponding 95% Confidence intervals (CI), showed that the growth in weight of the breastfed reference group was not different from the WHO Child growth standards. Between baseline and 13 weeks of age, the control group and experimental group showed a lower weight-for age z score than the WHO child growth standards for breastfed infants. For the ITT group the z scores of the experimental group were closer to the breast fed group, but the no statistical significant difference was suggested between the two groups (data not shown). At 17 weeks of age there were no statistically significant differences between the experimental or the control group and the WHO Child growth standards. In contrast, at 52 weeks of age the control group showed a significantly higher weight-for-age z-score compared to the WHO Child growth standards for breast fed infants. The experimental group was in particular at 52 weeks more similar to the breastfed reference group as indicated by the overlapping CI and for both groups there was no suggestion for a difference from the WHO Child growth standards. FIG. 1 shows the results obtained for the ITT dataset per age category (at birth, 17 and 52 weeks of age). The PP dataset showed similar results (data not shown).

[0126] Interestingly, although the effects were present both for girls and boys, the highest effect was observed for boys, both for PP and ITT group (data not shown).

[0127] At 52 weeks of age, the difference in weight between the study groups was compared using a PGC analysis, considering the overall study period (baseline until 52 weeks) and correcting for stratification factors. The results (in gram, for the overall period) as shown in Table 1 reveal that both for ITT and PP datasets the group that received the control formula has a higher weight, but no statistically significant different weight compared to the group receiving the experimental formula. However, the comparison of the control group with the breastfed reference group reveals a statistically significant higher weight in the control group for ITT and PP datasets, respectively (estimate of weight difference control vs. breastfed 551.09 g (p=0.0009) for ITT and 593.03 g (p=0.0009) for PP). The comparison of the experimental group with the breastfeed reference group showed higher weight in the experimental group, which was not statistically different for the PP dataset (estimate of weight difference 254.89 (p=0.1027). Compared to the control group, the experimental group was closer to the breast fed group.

TABLE-US-00001 TABLE 1 Differences in weight (grams) at 52 weeks between the study groups as by PGC analysis. ITT group PP group estimates of estimates of the difference in the difference in weight (grams) weight (grams) at 52 weeks at 52 weeks (p-value) (p-value) Experimental vs Control ?190.11 (0.2524) ?350.50 (0.0580) Experimental vs Breastfed 357.53 (0.0254) 254.89 (0.1027) Control vs Breastfed 551.09 (0.0009) 593.03 (0.0009) PGC analysis, considering the overall study period (baseline until 52 weeks) and correcting for stratification factors.

BMI

[0128] The median BMI for boys and for girls included in the three study groups was well within the z-score of ?1 and +1 of the WHO Child growth standard z-scores, for the ITT and the PP dataset (data not shown).

[0129] The mean BMI-for-age z scores and the corresponding 95% Confidence intervals, showed that the growth in BMI of the breastfed reference group was not different from the WHO Child growth standards. For the ITT group at 17 weeks of age there wase no statistical significant difference between the control group and the WHO Child growth standards, whereas the BMI z-score of the experimental group was lower and statistically significantly different compared to the WHO Child growth standards. In contrast, at 52 weeks of age the control group showed a significantly higher BMI-for-age z score compared to the WHO Child growth standards off breastfed infants The experimental group on the other hand was more similar to the breastfed reference group as indicated by the overlapping CI and for both groups there was no significant difference from the WHO Child growth standards. FIG. 2 shows the results obtained for the ITT dataset. The PP dataset showed similar results (data not shown). Interestingly, although the effects were present both for girls and boys, the highest effect was observed for boys, both for PP and ITT group (data not shown).

[0130] At 52 weeks of age, the difference in BMI between the study groups was compared using a PGC analysis, considering the overall study period (baseline until 12 months) and correcting for stratification factors. The results as shown in Table 2 reveal that both for ITT and PP datasets the group that received the control formula had the highest BMI, which was statistically significant different compared to the group receiving the experimental formula and compared to the breastfed reference group. The comparison of the experimental group with the breastfeed reference group showed higher, but not statistically significant different BMI in the experimental group, neither for the ITT nor the PP dataset.

TABLE-US-00002 TABLE 2 Differences in BMI (kg/m.sup.2) at 52 weeks between the study groups as by PGC analysis ITT group PP group estimates of estimates of the difference the difference in BMI at in BMI at 52 weeks (p- 52 weeks (p- value) value) Experimental vs. Control ?0.80 (0.0006) ?0.89 (0.0014) Experimental vs. Breastfed 0.41 (0.0682) 0.42 (0.0727) Control vs. Breastfed 1.22 (<.0001) 1.30 (<.0001) PGC analysis, considering the overall study period (baseline until 12 months) and correcting for stratification factors

Weight-for-Length

[0131] The median weight-for-length of boys and girls (included in the three study groups was well within the z-score of ?1 and +1 of the WHO Child growth standard z-scores, for the ITT and the PP dataset (data not shown).

[0132] In line with BMI-for age z-scores, none of the study group reveals a weight-for-length z-score at birth which was significantly different from the WHO Child growth standards. The mean weight-for-length z-scores of the experimental group was slightly lower and significantly different from the WHO Child growth at 17 weeks of age.

[0133] At 52 weeks of age there were not differences between the experimental group and the WHO Child growth standards anymore. On the contrary the mean weight-for-length z-scores of the control group at 52 weeks age was higher and significantly different from the WHO Child growth standards. FIG. 3 shows the results obtained for the ITT dataset. The PP dataset showed similar results (data not shown).

[0134] Interestingly, although the effects were present both for girls and boys, the highest effect was observed for boys, both for PP and ITT group (data not shown).

[0135] It should be noted that the difference between the control formula and the breastfed reference group are in line with the observations found in in the scientific literature when comparing breast fed infants with formula fed infants (Dewey et al., 1993, Am J Clin Nutr 57: 140-145). All mean values (for weight, BMI, weight for length, head circumference and skin folds) are well within the range of normal, healthy infants who are not at risk of obesity.

Skinfold Thickness and Head Circumference

[0136] Skinfold thickness is a non invasive measurement of body fat in infants and is an indication of the body development. At 52 weeks the biceps, triceps, suprailiac and subscapular skinfolds were determined, as well as the sum of these skinfolds. The skinfolds of the breast fed reference group (ITT) were lower in case of the biceps, triceps, suprailiac and sum of skinfolds, and were higher for the subscapular skinfold when compared with the control group. In all cases the skinfolds of the experimental group was intermediate, and closer to the breast fed reference group than the control group. The triceps and subscapular skinfold z score of the breast fed reference group was closest to the WHO Child growth standard of human milk infants, and also all the skinfolds in the experimental group were closer to the WHO Child growth standard than the control group.

[0137] Also the head circumference-by-age z-score of the experimental group (ITT) was very similar to that of the breastfed reference group, and similar to the WHO Child growth standard of breast fed infants. The z score of the control group (ITT) on the other hand was higher at 12 months of age than the other two groups and was significantly higher that the WHO Child growth standard.

[0138] These results are indicative for an effect of formula for infants on promoting a postnatal growth trajectory or body development in an infant towards a growth trajectory or body development which is similar to the growth trajectory or body development observed in human milk fed infants and/or on promoting a postnatal growth trajectory or body development in an infant towards a growth trajectory or body development which is closer to the optimal growth trajectory or body development of the WHO Child Growth Standards of human milk fed infants. Also these results are indicative for an effect of formula for infants on improving the postnatal growth trajectory or body development in an infant towards the growth trajectory or body development observed in human milk fed infants, in particular when compared to the growth trajectory or body development in infants fed infant formula of follow on formula not comprising milk fat or comprising between 15 to 20 wt. % palmitic acid based on total fatty acids, yet with less than 15% of the palmitic acid residues being in the sn-2 position of triglycerides. Further these results are indicative for an effect of formula for infants on promoting a balanced growth trajectory or body development in an infant and/or preventing or reducing the risk of an unbalanced growth trajectory or body development in an infant.