2 '-fucosyllactose and lactose-n-neotetraose for prevention and treatment of overweight

Abstract

The present invention relates to the use of 2 '-fucosyllactose and lactose-N-neotetraose for the treatment and / or weight reduction in a subject in need thereof. The invention also relates to pharmaceutical and food compositions containing these compounds.

Description

Technical Field

[0001] The present invention relates to the field of weight loss. In particular, the present invention relates to the use of two compounds, namely 2'-fucosyllactose and lacto-N-neotetraose, for reducing weight in a subject in need thereof. Background Art

[0002] Being overweight is an integral aspect of society in industrialized countries, which is often seen as not meeting 21st century standards of beauty. In 2019, about 65% of the adult population in the United States was considered overweight, while this rating applied to about 53% of the EU population. Meanwhile in China, about 50% of the adult population is considered above normal weight. What all these countries have in common, respectively, is that in recent years more and more children have become overweight and at risk of obesity.

[0003] Obesity is a risk factor associated with many different diseases that may lead to higher morbidity and ultimately higher mortality. Diseases commonly observed in obese individuals are diabetes (type 2), insulin resistance syndrome, cardiovascular disease, hypertension, atherosclerosis, congestive heart failure, stroke, gallbladder disease, osteoarthritis, sleep apnea, reproductive disorders (such as polycystic ovary syndrome). In addition, it is assumed that the higher incidence of cancers such as breast cancer and prostate cancer is associated with obesity.

[0004] Particularly in view of these health problems, it is generally desirable to reduce body weight. Attempts to achieve this goal by the individual themselves include adopting a diet that does not provide the body with sufficient calories, radically changing the nature of the food consumed, for example, omitting any meat or eating only after an extended period of time (e.g., only after 16 hours). All of these measures are cumbersome and unpopular with the individual, who must change his or her lifestyle.

[0005] In addition, the food industry has found this problem to be a promising new market and has proposed many different ingredients that help individuals maintain weight or even lose weight. These ingredients are agents that inhibit the deposition of fat in cells or inhibit the deposition of fat in cells by increasing the rate of lipid oxidation. Another area of ​​interest involves agents that reduce appetite.

[0006] In addition, the pharmaceutical industry has paid attention to this problem and provided drugs that block the intake of calories from food. A famous representative of the drugs is An agent that blocks lipase in the intestine. However, There are side effects, which in some cases lead to fluidization of stool. In addition, as a synthetic molecule, It has other side effects because it is not naturally designed to exist in the body.

[0007] There remains a need to provide new agents that are safe for consumption or administration and that are effective in reducing weight in overweight or obese individuals. Summary of the invention

[0008] In order to solve the above problems, the present invention provides 2'-fucosyllactose and / or lactose-N-neotetraose for treating or preventing overweight, obesity and / or any disease related thereto.

[0009] 2'-fucosyllactose and lactose-N-neotetraose are members of the polysaccharide family present in human milk, and are referred to as human milk oligosaccharides (HMO). 2'-fucosyllactose is the most common HMO, and may account for 30% of all HMO in breast milk. In general, 2'-fucosyllactose has a protective effect against infectious diseases by preventing toxins or pathogens from adhering to the intestinal epithelium, and is related to promoting infant growth. On the other hand, lactose-N-tetraose as another member of HMO is a complex sugar enzymatically synthesized by substrate lactose. It is biologically relevant in the early development of infant intestinal flora, and is currently regarded as a prebiotic, promoting the growth of healthy bacteria (such as Bifidobacterium longum subspecies infantis) in the intestinal microbiome.

[0010] In the course of extensive research leading to the present invention, the inventors investigated the ability of the nematode worm C. elegans to accumulate lipids upon exposure to different agents. Although this may seem simple, C. elegans is a well-established model for research, particularly in the functional characterization of novel gene functions and drug targets identified using proteomic and genomic techniques. The common inheritance of cellular complexity and disease pathways of C. elegans and higher organisms, along with the simplicity and cost-effectiveness of culture, undeniably puts this organism center stage as an effective in vivo model that is amenable to whole-organism screening and target validation.

[0011] It has now surprisingly been found that specific concentrations of 2'-fucosyllactose or lacto-N-neotetraose are effective in reducing lipid levels in such organisms. In fact, it has been found that the effectiveness of 2'-fucosyllactose is comparable to that of (2S)-1-{[(3S,4S)-3-hexyl-2-oxo-4-oxetanyl]methyl}dodecyl (2S)-2-formamido-4-methylpentanoate (also known by the trade name The effectiveness of lactose-N-neotetraose is about 70-90% of that of 1,2-diol (a compound sold and recognized as an effective drug for the treatment of obesity). In addition, it has been found that lactose-N-neotetraose is about 1,2-diol (a compound sold and recognized as an effective drug for the treatment of obesity). As a reference the efficiency is about 40-60%.

[0012] The present invention also relates to a composition containing an effective amount of 2'-fucosyllactose, lactose-N-neotetraose or a combination thereof for treating and / or preventing overweight, obesity and / or any related disorders in a subject in need thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 Shown is the fat-reducing effect of 2'-fucosyllactose at different concentrations in C. elegans.

[0014] Figure 2 Shown is the fat-reducing effect of lactose-N-neotetraose at different concentrations in Caenorhabditis elegans. DETAILED DESCRIPTION OF THE INVENTION

[0016] The present invention relates to 2'-fucosyllactose and / or lactose-N-neotetraose for use in treating or preventing overweight, obesity and / or related diseases in a subject. 2'-fucosyllactose was first found in human milk and isolated from said human milk in the 1950s, and methods for its isolation are well known in the art. Since human milk is not available in large quantities and its separation from complex mixed milk is also cumbersome, the compound can also be chemically synthesized, or can even be produced in recombinant Escherichia coli (E. coli) by known means.

[0017] The subject to which the compound is to be administered is a mammal, particularly a human or a pet.

[0018] The present invention further provides a composition comprising an effective amount of 2'-fucosyllactose and / or lactose-N-neotetraose, which is used to treat overweight, obesity and / or any related diseases in a subject. In the context of the present invention, the phrase effective amount should refer to the amount that brings the desired effect (i.e., interferes with lipid metabolism) when applied to a subject, so that an individual weight loss can be observed. In principle, it is envisioned that the daily dose of a person is about 10mg / kg body weight to about 90mg / kg body weight, preferably, about 15mg / kg body weight to about 65mg / kg body weight of 2'-fucosyllactose, more preferably, about 25mg / kg body weight to about 40mg / kg body weight. For lactose-N-neotetraose, it is envisioned that the daily dose is about 5mg / kg to about 50mg / kg, preferably, about 8mg / kg body weight to about 40mg / kg body weight, more preferably, about 12mg / kg body weight to about 20mg / kg body weight.

[0019] The composition proposed in the present invention may contain any of the corresponding compounds alone or in combination of both (2'-fucosyllactose and / or lacto-N-neotetraose).

[0020] The composition proposed in the present invention can be provided as a pharmaceutical composition or as a food composition.

[0021] When provided as a pharmaceutical composition, the composition may further comprise conventional agents designed to produce or shape the product, provide targeted release, or to protect the compound from environmental conditions to which it is exposed as it moves through the gastrointestinal tract until reabsorption. Non-limiting examples of these agents are fillers, solvents, emulsifiers, buffers, thickeners, coatings, disintegrants, lubricants, glidants, preservatives, reabsorption accelerators, and the like.

[0022] The pharmaceutical composition may be suitable for oral administration and may be in a form selected from the group comprising tablets, dragees, capsules, granules, powders, syrups, aqueous suspensions or aqueous solutions. Alternatively, the composition may be in the form of a depot.

[0023] The pharmaceutical composition can be combined with other known agents for reducing and / or preventing overweight and / or obesity and related diseases (such as 5-HTP, Lorcaserin and Bupropion / Naltrexone. Additional agents may be provided separately or may be included in the compositions described herein.

[0024] In the context of the present invention, the phrase related diseases includes any disease or condition for which overweight or obesity is a known risk factor. Non-limiting examples are metabolic syndrome, glycemia, type 2 diabetes, hypercholesterolemia, cardiovascular disease, hypertension, inflammation, hormonal changes, infertility, etc.

[0025] The composition proposed herein can also be in the form of food composition. In the context of the present invention, the term food composition includes any substance, additive or supplement of the meal itself, etc. Due to the properties of compound 2'-fucosyllactose and / or lactose-N-neotetraose, these two natural milk oligosaccharides, compounds or compounds can be mixed into any food material taken by an individual respectively, without producing the expected substantial side effect. Therefore, these compounds can be added to any conventional food product, such as juice, dairy products (such as milk, yogurt, kefir yogurt (kefir), cheese, ice cream, soft drinks, cakes) etc. from fruit or vegetable sources. The compound can also be formulated as a nutritional supplement containing concentrated form oligosaccharides, and therefore high doses allow it to be more highly absorbed. Similarly, food composition can also be designed for animals, such as pets, wherein the compound as shown herein is mixed into conventional pet food in any amount, provided that the physical appearance and taste of the pet food are not deteriorated.

[0026] According to one embodiment, the food composition as shown above can also be used together or separately with other known weight loss supplements or agents (such as or microcrystalline cellulose) supplement.

[0027] In addition, the present invention provides use of 2'-fucosyllactose and / or lacto-N-neotetraose and / or a composition containing these compounds for reducing the weight of non-obese people. Example

[0028] Example 1: Effect of 2'-fucosyllactose on fat reduction in Caenorhabditis elegans

[0029] Plates with a diameter of 100 x 15 mm were filled with NGM (nematode growth medium, as standard medium, hereinafter referred to as NG medium (Fisher Scientific)). Nile red (Sigma, St. Louis, MO, USA) was added to the solid surface of the NGM plate seeded at a final concentration of 0.05 μg / mL.

[0030] For each experiment, two NG medium plates were used, which had been supplemented with different amounts of 2'-fucosyllactose (i.e., 0.01 mg / mL, 0.1 mg / mL, 1 mg / mL, 10 mg / mL, and 30 mg / mL). Two plates with NG medium were used as negative controls. Two plates with The NG medium plate was used as a positive control.

[0031] Sixty synchronized worms at the young stage were added to each plate with E. coli strain OP50 as food source and cultured at 20 °C under aerobic conditions.

[0032] All worms were subjected to lipid staining following a previously described protocol (Martorell et al., 2016).

[0033] All nematodes from each plate were transferred to M9 buffer and fluorescence was measured using an FP-8300 system (JASCO Analytical Instrument, Easton, MD, USA). Fat deposition in worms was determined by one-way ANOVA test using Tukey's multiple comparison post-test.

[0034] Results are given as mean ± standard deviation. Statistical analysis was performed using GraphPad Prism 9 software. Statistical significance was indicated at 5%.

[0035] The results are shown in Table 1 and Figure 1 middle.

[0036] Table 1: Percentage of fat reduction at different doses of 2'-fucosyllactose (2'-FL) relative to the control feed (NG medium)

[0037] condition Fat loss relative to control (%) statistics Orlistat (6 μg / mL) 30.40 P<0.001 2'-FL (0.01 mg / L) -2.38 Not significant 2'-FL (0.1 mg / L) 11.37 P<0.05 2'-FL(1mg / L) 23.61 P<0.001 2'-FL(10mg / L) 25.35 P<0.001 2'-FL(30mg / L) 5.28 Not significant

[0038] The results showed that fat reduction was dose-dependent with the applied 2'-FL. The lower and higher doses did not produce functional effects, while the intermediate doses (0.1 mg / mL, 1 mg / mL, and 10 mg / mL, see Figure 1 , purple and dark green bars) significantly reduced the fat content of the nematodes, providing a 23.6% and 25.3% fat reduction compared to the negative control condition (NG medium). This effect was similar to that of the positive control orlistat (see Figure 1 , red bars), the positive control orlistat caused a 30.4% fat reduction compared to the negative control. Therefore, the most effective doses of 2'-FL, 1 mg / mL and 10 mg / mL, made their effectiveness about 78% and 83% of the effectiveness of the positive control orlistat.

[0039] Example 2: Effect of lactose-N-neotetraose on fat reduction in Caenorhabditis elegans

[0040] The assay of Example 1 was repeated, provided that lactose-N-neotetraose (doses 0.01 mg / mL, 0.1 mg / mL, 1 mg / mL, 10 mg / mL and 30 mg / mL) was used instead of 2-fucosyllactose.

[0041] The results are shown in Table 2 and Figure 2 middle.

[0042] Table 2: Percentage of fat reduction at different doses of lactose-N-neotetraose (LNnT) relative to the control feed (NG medium)

[0043] condition Fat loss relative to control (%) statistics Orlistate (6 μg / mL) 31.14 P<0.001 LNnT (0.01mg / L) -5.01 Not significant LNnT (0.1mg / L) -0.08 Not significant LNnT (1mg / L) 3.08 Not significant LNnT (10mg / L) 14.05 P<0.01 LNnT (30mg / L) -2.39 Not significant

[0044] The results showed that in the presence of 10 mg / mL lactose-N-neotetraose (see Figure 2 , dark green bars) had 14.05% less fat content than worms under negative control conditions (NGM). Orlistat again served as a positive control in this method, providing a 31.14% fat reduction effect (see Figure 2 , red bars). 10 mg / mL lactose-N-neotetraose provided an effect that was approximately 45% of that provided by the positive control.

Claims

1. 2'-fucosyllactose and / or lactose-N-neotetraose for use in preventing and / or treating overweight and / or obesity in a subject.

2. A composition comprising an effective amount of 2'-fucosyllactose and / or lacto-N-neotetraose for use in treating obesity and / or overweight in a subject.

3. A composition for use according to claim 2, comprising 2'-fucosyllactose and lacto-N-neotetraose.

4. A composition for use according to claim 2 or 3, wherein the composition is suitable for oral administration.

5. The composition for use according to claims 2 to 4, which is a pharmaceutical composition.

6. The composition for use according to claim 5, further comprising at least one carrier and / or pharmaceutically acceptable excipient.

7. A composition for use according to claim 3 or 4, in the form of tablets, capsules, powders, granules, syrups, aqueous suspensions or solutions.

8. A composition for use according to any one of claims 3 to 5, comprising an additional agent for reducing obesity and / or weight in a subject.

9. The composition for use according to claim 6, wherein the additional agent is 5-HTP, lorcaserin, and bupropion / naltrexone.

10. The composition for use according to claim 2, which is a food composition.

11. A composition for use according to claim 10, selected from the group comprising: fruit or vegetable juice, ice cream, infant formula, milk, yoghurt, cheese, fermented milk, milk powder, cereals, bakery products, products based on milk and / or cereals, nutritional supplements, soft drinks, dietary supplements or pet food.

12. The composition for use according to claim 10, wherein the composition is in the form of a food additive for a supplement.

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