Human milk oligosaccharides that can enhance bioavailability of the mineral zinc and uses thereof

Abstract

The present application provides human milk oligosaccharides capable of improving the bioavailability of mineral zinc and applications thereof. Specifically, the present application provides the use of lacto-N-neotetraose (LNnT) in the preparation of products capable of improving the bioavailability of mineral zinc. In the present application, lacto-N-neotetraose is used to improve the bioavailability of mineral zinc, especially for individuals lacking zinc, which can significantly improve the retention of zinc in the body and promote the digestion and absorption of zinc by individuals.

Description

Technical Field

[0001] This invention relates to a human milk oligosaccharide that can improve the bioavailability of the mineral zinc and its application, specifically the application of lactose-N-neotetrasaccharide (LNnT) in improving the bioavailability of the mineral zinc. Background Technology

[0002] The immune system is the body's natural defense mechanism, playing a vital role in fighting disease and infection, and regulating inflammation. Nutritional status and immune function are closely related and mutually influential. Malnutrition leads to weakened immune function, increasing susceptibility to infections and even tumors, while severe or chronic infections can trigger or worsen malnutrition. Proper nutrition is fundamental for maintaining normal immune function; nutrients are the material basis for the normal operation of the human immune system, playing a crucial role in both non-specific and specific immunity. Current research on nutrition and immunity mainly focuses on proteins, lipids, vitamins A, E, C, and D, iron, zinc, and selenium, especially the key role of micronutrients in the immune system.

[0003] Zinc is the second most abundant trace element in the human body, crucial for protein metabolism and synthesis, nucleic acid metabolism, and cell membrane stability. As a cofactor for over 300 coenzymes, zinc participates in enzyme activity and is essential for various cellular metabolic functions. Therefore, zinc plays a vital physiological role in growth and development, and the development and function of the immune system. Zinc deficiency is common in children, and their bioavailability is low. Zinc deficiency in humans can cause systemic symptoms such as short stature, underdeveloped genitals, iron-deficiency anemia, hepatosplenomegaly, and lethargy. Animal studies have shown that in zinc-deficient animal models during growth, immune organs atrophy, lymphocytes decrease, and immune function declines.

[0004] Zinc, a mineral that plays an important role in infant growth and development, cognitive development, and maintaining immune function, has been widely added to infant formula. However, the bioavailability of zinc added to infant formula is much lower than that of breast milk.

[0005] Therefore, solutions for zinc deficiency are needed, particularly in the area of ​​healthy eating for children, and efforts are needed to improve the bioavailability of zinc to prevent and / or improve immune reduction caused by zinc deficiency. Summary of the Invention

[0006] One object of the present invention is to provide a human milk oligosaccharide that can improve the bioavailability of zinc.

[0007] Another object of the present invention is to provide the application of human milk oligosaccharides in improving the bioavailability of the mineral zinc.

[0008] The inventors of this case discovered in their research that lactose-N-neotetrasaccharide (LNnT) can enhance the bioavailability of the mineral zinc, especially for zinc-deficient individuals, significantly increasing the retention rate of zinc in the body and promoting the digestion and absorption of zinc.

[0009] On the one hand, the present invention provides the application of lactose-N-neotetrasaccharide (LNnT) in the preparation of products that enhance the bioavailability of the mineral zinc.

[0010] Lacto-N-neotetraose (LNnT) is an important and core type 2 oligosaccharide in breast milk, composed of galactose (Gal), N-acetylglucosamine (GlcNAc), and glucose (Glc). LNnT possesses numerous physiological functions, including acting as a prebiotic to improve gut microbiota structure and achieving immunomodulatory functions by activating Th2 immunity. Currently, both the U.S. Food and Drug Administration (FDA) and the European Food Safety Authority (EFSA) have approved LNnT as a nutritional fortifier for infant formula. In 2023, LNnT was also approved for use as a food fortifier in infant formula in my country.

[0011] According to a specific embodiment of the present invention, the raw material for providing the lactose-N-neotetrasaccharide in the present invention can be a commercially available human milk oligosaccharide raw material with high purity. Unless otherwise specified, the amount or proportion of lactose-N-neotetrasaccharide in the composition of the present invention is based on the content of human milk oligosaccharide LNnT in the raw material.

[0012] According to a specific embodiment of the present invention, the application of lactose-N-neotetrasaccharide in the preparation of products that enhance the bioavailability of the mineral zinc is described. The present invention does not impose special requirements on the specific form of the product; for example, the product may be a nutritional supplement, dietary supplement, etc., and may be in the form of oral liquid, powder, capsule, etc. The product may be a general food, health food, or food for special medical purposes.

[0013] In some specific embodiments of the present invention, the product described herein is a dairy product, such as flavored milk or milk beverages, fermented milk or milk beverages, frozen drinks, milk powder, milk tablets, or cheese. The dairy product may be a food product intended for infants, children, adults, or the elderly.

[0014] In some specific embodiments of the present invention, the product described herein is infant formula, toddler formula, or infant formula for special medical purposes.

[0015] According to a specific embodiment of the present invention, the product contains 1.3-12.0 mg / 100g of zinc dry matter or 2-14 mg / L.

[0016] According to a specific embodiment of the present invention, the raw material for providing zinc in the product of the present invention includes one or more of zinc sulfate, zinc gluconate, zinc glycinate, zinc oxide, zinc lactate, zinc citrate, zinc chloride, zinc acetate, zinc carbonate, and zinc citrate trihydrate (zinc citrate (trihydrate)).

[0017] According to a specific embodiment of the present invention, the product of the present invention contains 130-500 mg / 100g of product dry matter or 0.2-0.6 g / L.

[0018] According to a specific embodiment of the present invention, the mass ratio of zinc to LNnT in the product of the present invention is 1:14 to 1:300.

[0019] On the other hand, the present invention also provides a method for improving the bioavailability of zinc in zinc-containing foods, the method comprising:

[0020] Control the content of zinc and LNnT in food so that the mass ratio of zinc to LNnT is 1:14 to 1:300.

[0021] According to a specific embodiment of the present invention, in the method for improving the bioavailability of zinc in zinc-containing foods, the food contains 1.3-12.0 mg / 100g of dry matter or 2-14 mg / L of zinc. Preferably, the food contains 2.6-12.0 mg / 100g of dry matter of zinc.

[0022] This invention can improve the bioavailability of zinc and can be used to prevent and / or improve the reduced immunity caused by zinc deficiency or insufficient zinc intake. Attached Figure Description

[0023] Figure 1 This shows the apparent mineral absorption rate of different groups in the first stage in specific embodiments of the present invention.

[0024] Figure 2 This shows the apparent mineral absorption rate of different groups in the second stage in specific embodiments of the present invention.

[0025] Figure 3 This shows the mineral retention rate of different groups in the first stage in specific embodiments of the present invention.

[0026] Figure 4 This shows the mineral retention rates of different groups in the second stage in specific embodiments of the present invention. Detailed Implementation

[0027] Before further describing the specific embodiments of the present invention, it should be understood that the protection scope of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the embodiments of the present invention are for describing specific embodiments and not for limiting the protection scope of the present invention.

[0028] When the embodiments give a numerical range, it should be understood that unless otherwise specified in the present invention, either of the two endpoints of each numerical range and any value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those skilled in the art of this technology. In addition to the specific methods, devices, and materials used in the embodiments, according to the knowledge of those skilled in the art of this technology and the description of the present invention, any methods, devices, and materials similar or equivalent to the methods, devices, and materials described in the embodiments of the present invention in the prior art can also be used to implement the present invention.

[0029] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt the conventional techniques in this technical field.

[0030] In each embodiment and experiment, the zinc and LNnT used are commercially available food-grade raw materials. Unless otherwise specified, the zinc and LNnT mentioned in the present invention are calculated based on the effective content of the active ingredient zinc element and LNnT in the raw materials.

[0031] Effect experiment of LNnT on improving the bioavailability of zinc

[0032] Grouping of experimental animals

[0033] In this study, through animal experiments, 3-week-old male Wistar rats were divided into a zinc-deficient group, a zinc-deficient + low-concentration LNnT group, and a zinc-deficient + high-concentration LNnT group.

[0034] The LNnT and ZnSO4·H2O used in the experiment are commercially available products. The SPF-grade male 3-week-old Wistar rats for animal experiments were purchased from Zhejiang Vital River Laboratory Animal Technology Co., Ltd., and the use license number is SCXK (Gan) 2023-0001.

[0035] Sixty male 3-week-old Wistar rats, weighing (131.0±11.6) g, were randomly divided into three groups of 20 rats each: Group 1 (control group), 30% zinc group (1.02 mg / kg per day zinc, approximately 30% of the normal daily zinc requirement); Group 2 (experimental group 1), 30% zinc + low-dose LNnT group (1.02 mg / kg per day zinc + 87.5 mg / kg per day LNnT); Group 3 (experimental group 2), 30% zinc + high-dose LNnT group (1.02 mg / kg per day zinc + 262.4 mg / kg per day LNnT). LNnT. Wistar rats were first acclimatized for one week, then divided into groups and fed a customized basal diet for growing rats (AIN-93G) for 28 days. The first group was fed a diet containing 10.2 ppm zinc, the second group a diet containing 10.2 ppm zinc and 875 ppm LNnT, and the third group a diet containing 10.2 ppm zinc and 2624 ppm LNnT. Apart from the diet, all groups of rats were kept under the same conditions: a temperature of 23–26°C, free access to water, daily weight recording, and weekly body length measurement. Wistar rats were housed at the Experimental Animal Science and Technology Center of Jiangxi University of Traditional Chinese Medicine.

[0036] Customized AIN-93G rat feed was purchased from Beijing Xiaoshu Youtai Biotechnology Co., Ltd.

[0037] Each group of rats was divided into two subgroups of 10 rats each. Each subgroup entered the metabolic cage to participate in the metabolic experiment on days 14-16 and 25-27. During this period, the food intake of each rat was recorded daily, and urine and feces were collected daily and stored at -80℃ for subsequent experiments.

[0038] Effect of LNnT on Zinc Bioavailability

[0039] Rat feces and urine obtained in the metabolic experiment were thawed at room temperature, weighed in a crucible, dehydrated and dried in a vacuum drying oven to constant weight, and weighed again. According to GB 5009.14-2017 "National Food Safety Standard - Determination of Zinc in Food", the samples were digested by dry ashing, and then the zinc content in the rat feces and urine of each group was determined by flame atomic absorption spectrometry.

[0040] Zinc bioavailability is characterized by apparent mineral uptake and mineral retention efficiency, and the formulas for calculating these two indicators are shown below:

[0041] Apparent mineral absorption rate (%) = (mineral intake - fecal mineral content) / mineral intake × 100.

[0042] Mineral retention efficiency (%) = (mineral intake - fecal mineral content - urinary mineral content) / mineral intake × 100.

[0043] Apparent mineral uptake rate refers to the percentage of minerals ingested by an animal that is absorbed. This indicator is primarily used to assess mineral absorption capacity, but it may be affected by factors such as endogenous mineral excretion.

[0044] Mineral retention efficiency refers to the ratio of the amount of minerals deposited in an animal's body to the amount ingested, usually expressed as a percentage. This indicator reflects the actual utilization efficiency of minerals in an animal's body and is an important parameter for measuring mineral bioavailability.

[0045] Data Analysis: All results were analyzed using WPS Office. Results are expressed as mean ± standard deviation (Mean ± SD). Two-way ANOVA and Tukey multiple comparison test were used to examine the differences between the three groups in the two stages. p < 0.05 was considered statistically significant. Graphs were plotted using GraphPad Prism 8.

[0046] Table 1 and Figure 1 Table 2 shows the apparent mineral absorption rate in the first stage for different groups. Figure 2 The apparent mineral absorption rates in the second phase were shown for different groups. In the zinc-deficient group, LNnT significantly improved the mineral absorption rate of zinc (p < 0.05).

[0047]

[0048]

[0049] Table 3 and Figure 3 Table 4 shows the mineral retention rates in the first stage for different groups. Figure 4 The second-stage mineral retention rates were shown for different groups. In the zinc-deficient group, LNnT significantly improved zinc retention (p < 0.05).

[0050]

[0051]

[0052] In summary, LNnT can improve the bioavailability of zinc, and this effect is more pronounced in the zinc-deficient group.

[0053] In addition, the effects of LNnT on the body length and weight of the test rats in this experiment are shown in Tables 5 and 6.

[0054]

[0055]

[0056] The above description of the embodiments is provided to enable those skilled in the art to understand and use the invention. It will be apparent to those skilled in the art that various modifications can be made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present invention is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the invention should be within the protection scope of the present invention.

Claims

1. Application of lactose-N-neotetrasaccharide in the preparation of products that enhance the bioavailability of the mineral zinc, wherein the product contains lactose-N-neotetrasaccharide in an amount of 130-500 mg / 100g of product dry matter or 0.2-0.6 g / L.

2. The application according to claim 1, characterized in that, The product is a nutritional supplement.

3. The application according to claim 1, characterized in that, The product is a general food, health food, or food for special medical purposes.

4. The application according to claim 1, characterized in that, The product in question is a dairy product.

5. The application according to claim 4, characterized in that, The dairy products mentioned are modified milk or modified milk beverages, fermented milk or fermented milk beverages, frozen drinks, milk powder, milk tablets or cheese.

6. The application according to claim 1, characterized in that, The product is infant formula or toddler formula.

7. The application according to claim 1, characterized in that, The product in question is an infant formula for special medical purposes.

8. The application according to any one of claims 1-7, characterized in that, The product contains 1.3-12.0 mg of zinc per 100g of dry matter or 2-14 mg / L.

9. The application according to claim 8, characterized in that, The zinc raw materials provided in the product include one or more of zinc sulfate, zinc gluconate, zinc glycinate, zinc oxide, zinc lactate, zinc citrate, zinc chloride, zinc acetate, zinc carbonate, and zinc citrate trihydrate.

10. The application according to claim 8, characterized in that, The mass ratio of zinc to lactose-N-neotetrasaccharide in the product is 1:14 to 1:

300.

11. A method for improving the bioavailability of zinc in zinc-containing foods, characterized in that, The method includes: The content of zinc and lactose-N-neotetrasaccharide in the food is controlled such that the mass ratio of zinc to lactose-N-neotetrasaccharide is 1:14 to 1:300, and the amount of lactose-N-neotetrasaccharide in the food is 130-500 mg / 100g of dry matter or 0.2-0.6 g / L.

12. The method according to claim 11, characterized in that, The food contains 1.3-12.0 mg of zinc per 100g of dry matter or 2-14 mg / L.

13. The method according to claim 12, characterized in that, The food contains 2.6-12.0 mg of zinc per 100g of dry matter.

Images