Extract of algae of the genus ulva for the treatment of disorders caused by a metabolic syndrome

Abstract

The present invention relates to an extract of algae of the genus Ulva and the use thereof in human and animal nutrition for the treatment of inflammation, preferably inflammation associated with a metabolic syndrome, and for the treatment of cognitive and / or emotional disorders caused by a metabolic syndrome. The present invention also relates to dietary supplements and food compositions comprising an extract of algae of the genus Ulva.

Description

Description Title: Ulva algae extract for the treatment of metabolic syndrome-induced disorders Technical field

[0001] The present invention relates to an extract of algae of the genus Ulva and its use in human and animal nutrition, for the treatment of disorders induced by a metabolic syndrome, preferably inflammation, and cognitive and / or emotional disorders. The present invention also relates to food supplements and food compositions comprising an extract of algae of the genus Ulva. Prior art

[0002] Genetic predisposition, sedentary lifestyle, dietary habits, environmental factors, and metabolic alteration are the main and converging factors in the onset of obesity, a well-known complex and multifactorial pathology that has reached pandemic proportions. The incidence of obesity has increased very rapidly over the last 30 years, suggesting that behavioral and environmental factors, particularly greater availability and consumption of high-fat foods (HFD), have strongly fueled this condition. Obesity induced by a high-fat diet is linked to several pathological conditions such as diabetes, cardiovascular diseases, hypertension, liver diseases, and some forms of cancer (colon, gallbladder, breast, etc.).

[0003] The common feature of all obesity-induced metabolic diseases is inflammation, particularly chronic low-grade inflammation that is closely linked to the pathophysiology of adipose tissue.

[0004] Indeed, obesity induced by a high-fat diet is characterized by low-grade inflammation, as well as insulin resistance, oxidative stress, and mitochondrial dysfunction.

[0005] This low-grade inflammation is distinguished from inflammation, in that low-grade inflammation is a low-noise, asymptomatic, and chronic intracellular pathophysiological mechanism. This chronic phenomenon results in cellular, tissue, organ, and functional damage. In contrast, inflammation refers to the adaptive physiological changes that aim to halt the progression of tissue damage, isolate and eliminate infectious agents, and activate the repair processes necessary to restore the organism's initial state (Baumann and Gauldie, 1994).

[0006] Cognitive and / or emotional disturbances are also induced by metabolic syndrome.

[0007] Many compositions have been developed to prevent or treat these pathologies. However, existing products or methods to meet these needs are limited by: - either the complexity, and therefore the cost of formulating these compositions comprising ingredients of very varied purity levels, - either the lack of factual demonstration of the anti-inflammatory activity or the neuro-protective activity of the extracts.

[0008] There is thus a need to develop molecules targeting inflammation in order to treat inflammation, preferably low-grade inflammation, and also to treat cognitive and / or emotional disorders induced by metabolic syndrome, in order to treat disorders induced by metabolic syndrome.

[0009] In this context, algae are of growing interest to this market, which is demanding new products of natural origin. Indeed, while they have been valued for decades, mainly as a source of food, for the texturizing properties of the polysaccharides they produce or, more recently, for their dermo-cosmetic properties, marine macroalgae, in particular, present a wide diversity of molecules that have still been little studied.

[0010] Over the period 1999-2015, 99 purified compounds from marine macroalgae with neuroprotective activity were identified. Of these compounds, 57 originate from brown macroalgae, 28 originate from red macroalgae, and only 14 originate from green macroalgae.

[0011] From the perspective of green marine macroalgae, there is a substantial body of literature highlighting the properties of compounds isolated from green algae or complex green algae extracts in the nutraceutical or pharmaceutical fields. However, no data on anti-inflammatory activities against low-grade inflammation, let alone in the context of metabolic syndrome, of these same compounds or extracts have been found.

[0012] It therefore appears that marine macroalgae, and in particular green algae, can still be considered to date as an under-exploited reservoir of anti-inflammatory compounds against low-grade inflammation and more generally for the treatment of disorders induced by metabolic syndrome. Summary of the invention

[0013] The present invention relates to an extract of algae of the genus Ulva, characterized in that: - the protein content is between 5 and 60% by mass, preferably between 15 and 50% by mass, preferably between 25 and 35% by mass, relative to the total dry mass of the extract, - the content of total phenolic compounds is between 0.5 and 40% by mass, preferably between 1 and 20% by mass, preferably between 1 and 10% by mass, relative to the total dry mass of the extract, - the total sugar content is between 0.5 and 80% by mass, preferably between 1 and 50% by mass, preferably between 5 and 15% by mass, relative to the total dry mass of the extract.

[0014] In one embodiment, the algae extract of the genus Ulva according to the invention has a protein content of between 28 and 32% by mass relative to the total dry mass of the extract.

[0015] In one embodiment, the algae extract of the genus Ulva according to the invention has a content of total phenolic compounds of between 2 and 6% by mass relative to the total dry mass of the extract.

[0016] In one embodiment, the algae extract of the genus Ulva according to the invention has a total sugar content of between 5 and 15% by mass relative to the total dry mass of the extract.

[0017] In one embodiment, the algae extract of the genus Ulva according to the invention is an algae extract of the genus Ulva selected from Ulva lactuca, Ulva rigida, Ulva conglobata, Ulva ohnoi, Ulva reticulata, Ulva prolifera, Ulva flexuosa and Ulva intestinalis.

[0018] According to one embodiment, the present invention relates to an extract of algae of the genus Ulva according to the invention for the treatment of disorders induced by metabolic syndrome.

[0019] The inventors have in fact demonstrated that an extract of algae of the genus Ulva has anti-inflammatory properties, preferentially against low-grade inflammation and helps protect against a deterioration in cognitive abilities and emotional alteration induced by metabolic syndrome.

[0020] The in vitro anti-inflammatory effect of the Ulva algae extract according to the invention was evaluated on a murine macrophage cell line Raw 264.7 provided by ATCC Cell (“American Type Culture Collection”): code ATCC-TIB-71. These are macrophages immortalized mice used as reference biological study models in metabolic diseases, to measure anti / pro-inflammatory activity.

[0021] Advantageously, the inventors have demonstrated that after 6 hours of pre-incubation with the extract according to the invention, the production of nitric oxide (NO) induced by LPS is drastically reduced, compared to inflammatory control conditions involving LPS alone. These results indicate that the extract according to the invention is very strongly anti-inflammatory because it induces a reduction in NO secretion close to that induced by dexamethasone, at very low concentration. Indeed, nitric oxide (NO) is a central mediator of the inflammatory process, which regulates the activity, growth and death of many types of immune and inflammatory cells. A reduction in NO secretion reflects anti-inflammatory activity.

[0022] In addition, the modulation of protein signaling pathways involved in inflammation by the extract according to the invention was evaluated. Advantageously, the inventors demonstrated that the extract according to the invention induces a drastic reduction in the expression of NLRP3 and iNOS proteins, involved in the inflammatory process in Raw 264.7 macrophages in culture.

[0023] To complete these tests, the modulation by the extract according to the invention of the secretion of pro-inflammatory cytokines in the extracellular medium was evaluated. Advantageously, the inventors demonstrated that the algae extract according to the invention induces, after 6 hours of pre-incubation, a drastic reduction in the secretion of TNF-a and IL-6 induced by LPS, reaching a level similar to that obtained in the presence of dexamethasone, a reference anti-inflammatory compound, even at the very low concentration of 0.1 pg / ml.

[0024] These results demonstrate the anti-inflammatory effect against chronic low-grade inflammation of the extract according to the invention, in a context of metabolic syndrome, with respect to the production of NO, the expression of key proteins involved in the signaling cascades linked to the inflammatory process (NLRP-3 and iNOS), and the secretion of pro-inflammatory cytokines. Thus, the present invention also relates to an extract of algae of the genus Ulva according to the invention for its use in the treatment of inflammation.

[0025] In one embodiment, the inflammation is inflammation induced by metabolic syndrome.

[0026] The inventors also evaluated the effect of the extract according to the invention on metabolic and behavioral alterations by evaluating the parameters metabolic (weight, fat mass and lean mass) and behavioral (anxiety-like behavior, memory capacities) in a model of metabolic syndrome.

[0027] The animals' anxiety-like behavior was assessed using the elevated plus maze test. The inventors demonstrated that supplementation with the extract according to the invention restores the animals' exploration time in the open arms (F (2, 20) = 7.376, p < 0.01). Supplementation with the extract according to the invention thus protects against the onset of anxiety-like disorders induced by the HFD diet.

[0028] The mice were also tested in the Morris water maze to assess the chronic effects of the treatment on memory. Interestingly, supplementation with the extract according to the invention restored the cognitive abilities of animals on the HFD diet. Indeed, they discriminated the target quadrant in a statistically significant manner compared to chance (p<0.05).

[0029] The inventors have thus demonstrated that the extract according to the invention protects against a deterioration in cognitive abilities and emotional alteration induced by metabolic syndrome.

[0030] Thus, the present invention also relates to an extract of algae of the genus Ulva according to the invention for its use in the treatment of cognitive and / or emotional disorders induced by metabolic syndrome.

[0031] According to one embodiment, the cognitive and emotional disorders are selected from hippocampal-dependent long-term memory disorders and anxiety-type disorders.

[0032] The present invention also relates to a food supplement or a food composition for humans or pets comprising an extract of algae of the genus Ulva according to the invention.

[0033] Indeed, the algae extract according to the invention can be used in human nutrition but also in animal nutrition. Brief description of the drawings

[0034] Other features, details and advantages will become apparent upon reading the detailed description below, and upon analyzing the attached drawings, in which: Fig. 1

[0035] [Fig. 1] shows the effect of the P fraction on the viability of murine Raw 264.7 macrophages in culture, after 24 h of incubation (A) or after 6 h of pre-incubation followed by 18 h of incubation in the presence of LPS (1 pg / ml) (B). The results express the percentage Relative mean of viable macrophages compared to control conditions (100%) ± SEM. Significant differences obtained between samples and control conditions (A) or between samples and conditions with LPS alone (B) are indicated by (n = 4): ns (not significant: p>0.05) (One-WAY ANOVA). Fig. 2

[0036] [Fig. 2] shows the effect of the P fraction on nitric oxide (NO) secretion by murine Raw 264.7 macrophages in culture, after 24 h of incubation (A) or after 6 h of pre-incubation followed by 18 h of incubation in the presence of LPS (1 pg / ml) (B). The results express the concentration of NO secreted in the culture medium ± SEM. The significant differences obtained between samples and control conditions (A) or between samples and conditions with LPS alone (B) are indicated by (n = 4): ns (not significant: p>0.05), *** (p<0.001) and **** (p<0.0001) (One-WAY ANOVA). Fig. 3

[0037] [Fig. 3] shows the effect of the P fraction on the respective expression of NLRP3 (A) and iNOS (B) proteins, by Raw 264.7 murine macrophages in culture, after 6 h of pre-incubation and then 18 h of incubation in the presence of LPS. The results express the level of expression of the target proteins ± SEM, reported to the level of expression of p-actin, as a housekeeping protein. The significant differences obtained between the samples and the conditions with LPS alone are indicated by (n = 6): ns (not significant: p>0.05), * (p<0.05), ** (p<0.01 ) and *** (p<0.001 ) (One-WAY ANOVA). Fig. 4

[0038] [Fig. 4] shows the effect of the P fraction on the secretion of TNF-a and IL-6 by murine Raw 264.7 macrophages in culture, after 24 h of incubation (A, C) or after 6 h of pre-incubation followed by 18 h of incubation in the presence of LPS (B, D). The results express the concentration of TNF-a and IL-6 secreted in the culture medium ± SEM. The significant differences obtained between samples and control conditions (A, C) or between samples and conditions with LPS alone (B, D) are indicated by (n = 4): ns (not significant: p>0.05), * (p<0.05), ** (p<0.01) and *** (p<0.001) (One-WAY ANOVA). Fig. 5

[0039] [Fig. 5] shows the effect of P fraction supplementation on metabolic parameters of obese and diabetic animals fed with HFD diet: body weight (A), lean mass (B) and fat mass (C) of C57BL / 6J mice fed with HFD or standard A04 diet for 16 weeks and supplemented for the last 4 weeks with the P fraction solution at 1 mg / ml in drinking water or with water (controls). Significant differences are indicated by (n = 12 / group): ns (not significant: p>0.05), ** (p<0.01), *** (p<0.001) and **** (p<0.0001) (One-WAY ANOVA + Tuckey post-hoc test). Fig. 6

[0040] [Fig. 6] shows the effect of P fraction supplementation on the exploration time of the anxiety-inducing open arms (OA) of the elevated plus maze, in obese and diabetic animals fed with a HFD diet: C57BL / 6J mice fed with a HFD or standard A04 diet for 16 weeks and supplemented for the last 4 weeks with the P fraction solution at 1 mg / ml in drinking water or with water (controls). Significant differences are indicated by (n = 12 / group): * (p<0.05) and ** (p<0.01) (One-WAY ANOVA + Tuckey post-hoc test). Fig. 7

[0041] [Fig. 7] shows the schematic representation of the Morris water maze behavioral test protocol. Fig. 8

[0042] [Fig. 8] shows the effect of P fraction supplementation on memory in obese and diabetic animals fed with HFD diet. Memory is evaluated according to the time spent in the quadrant opposite the platform (Opp (E)), the quadrants adjacent to the platform (Adj (N or S)) or in the target quadrant where the platform is located (Target (W)) by C57BL / 6J mice fed with HFD or standard A04 diet for 16 weeks, and supplemented during the last 4 weeks with P fraction at 1 mg / ml in drinking water or with water (controls). Significant differences between the values ​​obtained and chance (indicative value of 25%) are indicated by (n = 12 / group): * (p<0.05) and *** (p<0.001) (One-WAY ANOVA + Tuckey post-hoc test). Detailed Description Extract of algae of the genus Ulva

[0043] The present invention relates to an extract of algae of the genus Ulva characterized in that: - the protein content is between 5 and 60% by mass, preferably between 15 and 50% by mass, preferably between 25 and 35% by mass, relative to the total dry mass of the extract, - the content of total phenolic compounds is between 0.5 and 40% by mass, preferably between 1 and 20% by mass, preferably between 1 and 10% by mass, relative to the total dry mass of the extract, - the total sugar content is between 0.5 and 80% by mass, preferably between 1 and 50% by mass, preferably between 5 and 15% by mass, relative to the total dry mass of the extract. Content in

[0044] According to one embodiment, the algae extract of the genus Ulva is characterized by a protein content of between 5 and 60% by mass relative to the total dry mass of the extract, preferably between 25 and 35% by mass relative to the total dry mass of the extract and preferably, the protein content is between 28 and 32% by mass, more preferably approximately 30% by mass relative to the total dry mass of the extract.

[0045] Protein content may be measured by any method known to those skilled in the art. Illustrative methods include the methods of Lowry (Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265-75), Smith (Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, et al. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76-85) and Kjeldahl (Kirk PL. Kjeldahl Method for Total Nitrogen. Anal Chem. 1950 Feb 15;22(2):354-8). These protocols may be used independently and alternatively to measure protein content. Total phenolic compound content

[0046] According to one embodiment, the algae extract of the genus Ulva is characterized by a content of total phenolic compounds of between 0.5 and 40% by mass relative to the total dry mass of the extract, preferably between 1 and 10% by mass relative to the total dry mass of the extract and preferably, the content of total phenolic compounds is between 2 and 6% by mass, more preferably approximately 4% by mass relative to the total dry mass of the extract.

[0047] The term "total phenolic compounds" refers to all secondary metabolites characterized by the presence of an aromatic cycle carrying free hydroxyl groups or those bound to a carbohydrate, present in all parts of higher plants, the most represented being anthocyanins, flavonoids and tannins.

[0048] The content of total phenolic compounds may be measured by any method known to those skilled in the art. For illustration purposes, the method of Singleton and Rossi (Singleton VL, Rossi JA. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am J Enol Vitic. 1965 Jan 1;16(3):144) may be cited, with some modifications. Total sugar content

[0049] According to one embodiment, the algae extract of the genus Ulva is characterized by a total sugar content of between 0.5 and 80% by mass relative to the total dry mass of the extract, preferably between 5 and 15% by mass relative to the total dry mass of the extract and preferably, the total sugar content is approximately 10% by mass, relative to the total dry mass of the extract.

[0050] “Total sugars” means all carbohydrates: mono-, oligo-, polysaccharides and their derivatives.

[0051] The total sugar content may be measured by any method known to those skilled in the art. For example, the phenol / sulfuric acid mixture method introduced by Dubois (Dubois M, Gilles K, Hamilton JK, Rebers PA, Smith F. A Colorimetric Method for the Determination of Sugars. Nature. 1951 Jul;168(4265):167-167) is an example.

[0052] According to a preferred embodiment, the algae extract of the genus Ulva is characterized in that: - The protein content is approximately 30% by mass relative to the total dry mass of the extract, - The content of total phenolic compounds is approximately 4% by mass relative to the total dry mass of the extract, - The total sugar content is approximately 10% by mass relative to the total dry mass of the extract.

[0053] As used herein, the term "about," when referring to a number or numerical range, means that the number or numerical range in question is an approximation within the limits of experimental variability (or within the limits of statistical experimental error), and thus that the number or numerical range may vary between 1% and 15% of the stated number or numerical range. For example, the use of "about X" encompasses + / -1 percent, 2 percent, 3 percent, 4 percent, 5 percent, 6 percent, 7 percent, 8 percent, 9 percent, 10 percent, 11 percent, 12 percent, 13 percent, 14 percent, and 15 percent of the value X. Algae of the genus Ulva

[0054] The algae of the genus Ulva are multicellular green algae distributed in the oceans with about 400 species.

[0055] According to one embodiment, the algae of the genus Ulva is chosen from all the species of the genus Ulva, preferably from the group consisting of the species Ulva lactuca, Ulva rigida, Ulva fasciata, Ulva pertusa, Ulva californica, Ulva gigantea, Ulva pseudocurvata, Ulva conglobata, Ulva armoricana, Ulva fasciculata, Ulva ohnoi, Ulva pseudolinza, Ulva reticulata, Ulva rotundata, Ulva elegans, Ulva prolifera, Ulva scandinavica, Ulva umbilicata, Ulva flexuosa, Ulva clathrata, Ulva compressa and Ulva intestinalis.

[0056] According to a preferred embodiment, the algae of the genus Ulva is chosen from Ulva lactuca, Ulva rigida, Ulva conglobata, Ulva ohnoi, Ulva reticulata, Ulva prolifera, Ulva flexuosa and Ulva intestinalis.

[0057] According to one embodiment, the algae of the genus Ulva is Ulva lactuca or Ulva rigida.

[0058] According to one embodiment, the algae is a mixture of algae of the genus Ulva lactuca and Ulva rigida.

[0059] According to one embodiment, the algae extract of the genus Ulva according to the invention can be obtained by the process described below. Disorders induced by metabolic syndrome

[0060] The present invention also relates to an extract of algae of the genus Ulva according to the invention for its use in the treatment of disorders induced by metabolic syndrome.

[0061] The term "metabolic syndrome" usually refers to energy metabolism disorders (fasting hyperglycemia, insulin resistance, hypertension, hypertriglyceridemia, visceral fat accumulation, low plasma HDL cholesterol levels) associated with obesity or overweight. Metabolic syndrome is defined as a combination of three or more of the abnormalities listed above.

[0062] Typically, metabolic syndrome induces low-grade inflammation and also results in cognitive impairment and / or emotional impairment.

[0063] Thus, according to one embodiment, the disorders induced by a metabolic syndrome are chosen from inflammation, preferably low-grade inflammation or inflammation induced by a metabolic syndrome and cognitive and / or emotional disorders.

[0064] The present invention also relates to a method for the treatment of disorders induced by metabolic syndrome, comprising administering an algal extract according to the invention to a mammal in need thereof.

[0065] The present invention also relates to the use of an algae extract according to the invention for obtaining a medicament intended for the treatment of disorders induced by metabolic syndrome.

[0066] The present invention also relates to a pharmaceutical or veterinary composition comprising an algae extract according to the invention for its use in the treatment of disorders induced by metabolic syndrome. Inflammation

[0067] Thus, the present invention relates to an extract of algae of the genus Ulva according to the invention for its use in the treatment of inflammation.

[0068] The inventors have advantageously demonstrated, on reference models in metabolic diseases, the anti-inflammatory effect of the extract according to the invention with regard to the production of NO, the expression of key proteins involved in the signaling cascades linked to the inflammatory process (NLRP-3 and iNOS), and the secretion of pro-inflammatory cytokines.

[0069] This anti-inflammatory effect is an effect against the inflammation induced by metabolic syndrome.

[0070] Thus, according to one embodiment, the present invention relates to an extract of algae of the genus Ulva according to the invention for its use in the treatment of inflammation induced by metabolic syndrome.

[0071] In the context of metabolic syndrome, we then speak of low-grade inflammation and not inflammation.

[0072] Inflammation is described as the succession of changes occurring in living tissue when it is injured (Punchard et al., 2004). The inflammatory response corresponds to the local component of a set of host defense mechanisms grouped under the name of acute phase reaction. This term refers to the adaptive physiological changes that aim to stop the progression of tissue damage, isolate and eliminate infectious agents and activate the repair processes necessary to restore the initial state of the organism (Baumann and Gauldie, 1994).

[0073] However, there is also low-grade inflammation corresponding to a low-level, asymptomatic, and chronic intracellular pathophysiological mechanism. This chronic phenomenon causes cellular, tissue, organic, and functional damage. This low-grade inflammation is present in many pathologies, particularly in the context of metabolic syndrome, where it contributes to the establishment of alterations induced by the syndrome.

[0074] Low-grade inflammation is therefore distinguished from inflammation.

[0075] According to one embodiment, the present invention relates to an extract of algae of the genus Ulva according to the invention for its use in the treatment of low-grade inflammation.

[0076] According to one embodiment, the present invention relates to an extract of algae of the genus Ulva according to the invention for its use in the treatment of low-grade inflammation induced by metabolic syndrome.

[0077] According to one embodiment, the present invention relates to an extract of algae of the genus Ulva according to the invention for its use for the improvement of at least one symptom or disorder associated with a metabolic syndrome.

[0078] According to one embodiment, the present invention relates to an extract of algae of the genus Ulva according to the invention for its use in the treatment of inflammation induced by obesity or overweight.

[0079] According to one embodiment, the present invention relates to an extract of algae of the genus Ulva according to the invention for its use in the treatment of low-grade inflammation induced by obesity or overweight.

[0080] Obesity and overweight can be determined, among other things, by calculating the body mass index (BMI). BMI is obtained by calculating the ratio between weight and height squared. For example, a person measuring 1.62 m and weighing 55 kg has a BMI equal to 20 (i.e.: 55 / 1.62 2 ). Generally speaking, a person is said to have a normal weight when their BMI is between 18 and 25 kg / m 2 , is overweight when their BMI is between 25 and 30 kg / m 2 , and is obese if their BMI exceeds 30 kg / m 2 The present invention also relates to a method for the treatment of inflammation, comprising administering an algal extract according to the invention to a mammal in need thereof.

[0081] The present invention also relates to a method for the treatment of inflammation induced by metabolic syndrome, comprising administering an algal extract according to the invention to a mammal in need thereof.

[0082] The present invention also relates to a method for the treatment of low-grade inflammation induced by metabolic syndrome, comprising administering an algal extract according to the invention to a mammal in need thereof.

[0083] The present invention also relates to a method for improving at least one symptom or disorder associated with metabolic syndrome, comprising administering an algal extract according to the invention to a mammal in need thereof.

[0084] The present invention also relates to a method for the treatment of inflammation induced by obesity or overweight, comprising administering an algal extract according to the invention to a mammal in need thereof.

[0085] The present invention also relates to a method for the treatment of low-grade inflammation induced by obesity or overweight, comprising administering an algal extract according to the invention to a mammal in need thereof.

[0086] The present invention also relates to the use of an algae extract according to the invention for obtaining a medicament intended for the treatment of inflammation.

[0087] The present invention also relates to the use of an algae extract according to the invention for obtaining a medicament intended for the treatment of inflammation induced by metabolic syndrome.

[0088] The present invention also relates to the use of an algae extract according to the invention for obtaining a medicament intended to improve at least one symptom or disorder associated with metabolic syndrome.

[0089] The present invention also relates to the use of an algae extract according to the invention for obtaining a medicament intended for the treatment of inflammation induced by obesity or overweight.

[0090] The present invention also relates to the use of an algae extract according to the invention for obtaining a medicament intended for the treatment of low-grade inflammation induced by obesity or overweight,

[0091] The present invention also relates to a pharmaceutical or veterinary composition comprising an algae extract according to the invention for its use in the treatment of inflammation.

[0092] The present invention also relates to a pharmaceutical or veterinary composition comprising an algae extract according to the invention for its use in the treatment of inflammation induced by metabolic syndrome.

[0093] The present invention also relates to a pharmaceutical or veterinary composition comprising an algae extract according to the invention for its use in improving at least one symptom or disorder associated with a metabolic syndrome.

[0094] The present invention also relates to a pharmaceutical or veterinary composition comprising an algae extract according to the invention for its use in the treatment of inflammation induced by obesity or overweight.

[0095] The present invention also relates to a pharmaceutical or veterinary composition comprising an algae extract according to the invention for its use in the treatment of low-grade inflammation induced by obesity or overweight. Treatment of cognitive and / or emotional disorders induced by metabolic syndrome.

[0096] The inventors have in fact demonstrated that the extract according to the invention protects against a deterioration in cognitive abilities and / or emotional alteration induced by metabolic syndrome.

[0097] Indeed, it has been demonstrated that the extract according to the invention protects against the onset of anxiety-type disorders induced by the HFD diet and that it allows the cognitive abilities of animals fed with an HFD diet to be restored.

[0098] Thus, the present invention also relates to an algae extract according to the invention for the treatment of cognitive and / or emotional disorders induced by metabolic syndrome.

[0099] According to one embodiment, the present invention also relates to an algae extract according to the invention for the treatment of cognitive and / or emotional disorders induced by obesity or overweight.

[0100] Cognitive and emotional disorders include hippocampal-dependent long-term memory disorders and anxiety-type disorders.

[0101] According to one embodiment, the present invention relates to an algae extract according to the invention for the treatment of hippocampo-dependent long-term memory disorders and / or anxiety-type disorders.

[0102] According to one embodiment, the present invention relates to an algae extract according to the invention for the treatment of hippocampo-dependent long-term memory disorders and / or anxiety-type disorders, induced by metabolic syndrome.

[0103] According to one embodiment, the present invention relates to an algae extract according to the invention for the treatment of hippocampo-dependent long-term memory disorders and / or anxiety-type disorders, induced by obesity or overweight.

[0104] Advantageously, the extract according to the invention restores episodic memory, hippocampus-dependent spatial working memory, affected during the development of metabolic syndrome, this metabolic disease characterized by the establishment of inflammation and peripheral and central insulin resistance.

[0105] Thus, according to one embodiment, the present invention relates to an algae extract according to the invention for the restoration of episodic memory, hippocampus-dependent spatial working memory.

[0106] The present invention also relates to a method for treating cognitive and / or emotional disorders induced by metabolic syndrome comprising the administration of an algae extract according to the invention to a mammal in need thereof.

[0107] The present invention also relates to the use of an algae extract according to the invention for obtaining a medicament intended for the treatment of cognitive and / or emotional disorders induced by metabolic syndrome.

[0108] The present invention also relates to a pharmaceutical or veterinary composition comprising an algae extract according to the invention for its use in the treatment of cognitive and / or emotional disorders induced by metabolic syndrome.

[0109] The extract according to the invention may be present in the form of a pharmaceutical or veterinary composition, a food supplement, a food composition for humans or animals as defined below.

[0110] Thus, the present invention also relates to an algae extract according to the invention characterized in that the algae extract is present in the form of a pharmaceutical composition, a veterinary composition, a food supplement or a food composition for humans or for pets. Treatment

[0111] The terms "treatment" or "treatment method" are not absolute terms and, when applied to disorders induced by metabolic syndrome in preference to inflammation or cognitive and / or emotional disorders induced by metabolic syndrome, designate a procedure or course of action designed, even with a low probability of success, to induce an overall beneficial effect such as delaying the onset of the pathology, or reducing the severity of one or more symptoms. Typically, in the case of inflammation induced by metabolic syndrome, treatment may be understood to mean decreasing the low-grade inflammatory response. Typically, in the case of inflammation induced by metabolic syndrome, treatment may be understood to mean decreasing cognitive and / or emotional disorders.Typically, in the case of inflammation induced by metabolic syndrome, treatment can be understood as improving memory capacity and / or reducing anxiety.

[0112] The terms "metabolic syndrome-induced" and "metabolic syndrome-associated" can be used interchangeably.

[0113] According to one embodiment, the invention relates to the use of an algae extract according to the invention for the improvement of at least one symptom associated with a metabolic syndrome, in which said improvement comprises the reduction of low-grade inflammation, the reduction of cognitive and / or emotional disorders, the improvement of memory capacities and / or the reduction of anxiety.

[0114] Whether for the treatment of inflammation or the treatment of cognitive and / or emotional disorders induced by metabolic syndrome, the mammal is chosen from humans and pets.

[0115] The algae extract according to the invention can in fact be used in human nutrition but also in animal nutrition.

[0116] According to one embodiment, the mammal is a human.

[0117] According to another embodiment, the mammal is a pet such as a dog or a cat.

[0118] According to one embodiment, the algae extract according to the invention may be administered in the form of a pharmaceutical or veterinary composition comprising, in addition to the algae extract, a pharmaceutically acceptable excipient.

[0119] “Pharmaceutically acceptable” means a substance that is not biologically or otherwise undesirable, i.e., can be incorporated into a pharmaceutical composition administered to a patient or animal without causing undesirable biological effects or without interacting deleteriously with any of the other components of the composition in which it is contained, for example by inhibiting or diminishing the anti-inflammatory properties of the seaweed extract.

[0120] Typically, the pharmaceutically acceptable excipient may be selected from a diluent, a disintegrant, a binder, a glidant, a lubricating agent, a wetting agent, a buffering agent, a suspending agent, an adjuvant, an emulsifier, an absorbent, a preservative, a surfactant, a sweetening agent, an antioxidant, or a mixture thereof. These excipients are for example described in "The Science and Practice of Pharmacy 1995, edited by EW Martin, Mack Publishing Company, 19th edition, Easton, Pa."

[0121] The amount of the algae extract according to the invention in the compositions may vary so as to administer an effective amount of the algae extract to obtain the desired therapeutic response for a particular mammal.

[0122] An "effective amount" or "therapeutically effective amount" of a compound means a non-toxic but sufficient amount of the compound to provide the desired effect.

[0123] Typically, the amount administered or dose depends on the activity of the algae extract according to the invention, the route of administration, the severity of the pathology, as well as the state of health and medical history of the mammal treated, various factors such as body weight, diet, and the possible combination with other therapeutic agents. However, it is within the skill of the person skilled in the art to determine the appropriate dosage and to initiate treatment at a dosage lower than that required to obtain the desired therapeutic effect and then to gradually increase the dose until the desired effect is obtained.

[0124] The algae extract can be administered orally. Typically, the pharmaceutical or veterinary compositions can be presented in the form of a food supplement for humans or pets or food compositions for humans or pets.

[0125] The present invention also relates to the use of a pharmaceutical or veterinary composition for its use in the therapeutic applications as described above. Process for obtaining the algae extract of the genus Ulva according to the invention

[0126] Advantageously, the algae extract according to the invention can be obtained according to the steps of the process described in patent FR 2 998 894 from the company SEPROSYS, which describes a process for treating algae comprising: - a grinding stage, - diffusion of the algae to be treated into the water, - filtration of the pulps recovered after the diffusion stage so as to obtain a pressing juice on the one hand and pressing pulps on the other hand, - ultrafiltration of the pressing juice in order to obtain a retentate on the one hand and a permeate on the other.

[0127] The inventors have advantageously discovered that the permeate obtained by implementing such a process on algae of the genus Ulva exhibited anti-inflammatory properties.

[0128] Preferably, the permeate presents anti-inflammatory properties in a context of metabolic syndrome.

[0129] Thus, the present invention also relates to a method for obtaining an extract of algae of the genus Ulva comprising the steps: - crushing algae - algae extraction - filtration in order to obtain an algae extract with anti-inflammatory properties.

[0130] The present invention also relates to a method for obtaining an extract of algae of the genus Ulva comprising the steps: - crushing algae - algae extraction - filtration in order to obtain an algae extract with anti-inflammatory properties in the context of metabolic syndrome.

[0131] The term “algae extract with anti-inflammatory properties” means an algae extract that can: - reduce the production of NO, or - reduce the expression of NLRP3 and iNOS proteins, or - reduce the secretion of TNF-a and IL-6.

[0132] Preferably, the anti-inflammatory properties are anti-inflammatory properties in a context of metabolic syndrome. Typically, these parameters can be measured on reference models in metabolic diseases, such as a murine macrophage cell line Raw 264.7 provided by ATCC Cell (“American Type Culture Collection”): code ATCC-TIB-71.

[0133] According to one embodiment, the algae grinding step makes it possible to obtain algae particles with a diameter of less than 10 mm, preferably less than 5 mm, preferably less than 2 mm.

[0134] Typically, the algae can be ground in demineralized water, at a temperature between 25 and 100°C, preferably between 40 and 90°C, preferably between 55 and 75°C, and preferably at a temperature of approximately 65°C.

[0135] Any utensil known to those skilled in the art can be used to grind the algae, typically a knife grinder.

[0136] According to one embodiment, the method may comprise a step prior to grinding, of washing the algae, typically in raw water for 10 min at room temperature, followed by spinning.

[0137] According to one embodiment, the algae extraction or diffusion step is carried out at a temperature between 25 and 100°C, preferably between 40 and 90°C and preferably at a temperature of approximately 80°C.

[0138] According to one embodiment, the duration of the algae extraction or diffusion step is between 15 min and 5 h, preferably between 1 h and 4 h, preferably between 1 h and 3 h. According to a preferred embodiment, the duration of the extraction step is approximately 2 h.

[0139] According to one embodiment, the extraction step is carried out in a reduced volume of water.

[0140] According to one embodiment, the extraction is carried out under constant stirring. By way of illustration, the extraction is carried out under constant stirring using a paddle stirrer, at a rotation speed of 10 rpm.

[0141] The extraction stage produces an aqueous extract and seaweed pulp.

[0142] According to one embodiment, the aqueous extract obtained following the extraction is filtered.

[0143] According to one embodiment, the filtration steps may be carried out according to the method described in patent FR 2 998 894 from the company SEPROSYS. Thus, the method comprises the steps: - filtration of the pulps recovered after the diffusion stage, so as to obtain a pressing juice on the one hand and pressing pulps on the other hand, - ultrafiltration of the pressing juice in order to obtain a retentate on the one hand and a permeate on the other.

[0144] Alternatively, the following filtration steps can be carried out after the extraction step.

[0145] Typically, the aqueous extract obtained after the extraction step can undergo a first filtration step and be filtered using a vibrating sieve with a cut-off threshold of 50 μm. This filtration step of the aqueous extract makes it possible to eliminate algae residues.

[0146] According to one embodiment, the pulps obtained after the extraction step are pressed.

[0147] This step advantageously makes it possible to obtain a pressed juice on the one hand and pressed pulp on the other hand, and to extract as much juice as possible. According to one embodiment, the pressed juice is added to the aqueous extract.

[0148] According to one embodiment, the method comprises, following the first filtration step, three additional filtration steps.

[0149] According to one embodiment, the method according to the invention comprises a first microfiltration step.

[0150] As an illustration, a plate filter type microfiltration unit equipped with two filters of 1.2 pm then 0.8 pm (3M, 40*40, pore size 15 HN and 50 HN) can be used.

[0151] The microfiltration stage makes it possible to obtain, from the pressing juice, a retentate on the one hand and a permeate on the other.

[0152] The permeate obtained following the microfiltration step can be filtered via an ultrafiltration unit.

[0153] Thus, according to one embodiment, the method according to the invention comprises an ultrafiltration step.

[0154] According to one embodiment, the method according to the invention comprises a step of ultrafiltration of the permeate obtained following microfiltration so as to obtain a retentate on the one hand and a permeate on the other hand.

[0155] According to one embodiment, the ultrafiltration step is carried out between 60 and 100°C, preferably at approximately 80°C, at a pressure of between 2 and 10 bars, preferably at approximately 5 bars and at a circulation flow rate of between 300 l / h and 600 l / h, preferably at 450 l / h.

[0156] According to one embodiment, the ultrafiltration step is carried out until a retentate of approximately 4 Bx is obtained.

[0157] 4 Bx or Brix corresponds to approximately 4% of dry matter measured by the refractive index and expressed in degrees brix sucrose scale.

[0158] The retentate can be withdrawn from the unit and the permeate can be recovered and then osmosed in a reverse osmosis unit.

[0159] For illustration purposes, ultrafiltration can be carried out on an ultrafiltration unit equipped with a Kerasep KBW 15 kDa membrane (Novasep Process).

[0160] According to one embodiment, the method according to the invention comprises a reverse osmosis step.

[0161] According to one embodiment, the method according to the invention comprises a step of reverse osmosis of the ultrafiltration permeate.

[0162] According to one embodiment, the reverse osmosis step is carried out between 20 and 50°C, preferably between 25 and 40°C and preferably, the reverse osmosis step is carried out at approximately 30°C, at a pressure of between 10 and 50 bars, preferably between 20 and 30 bars and at a circulation flow rate of between 80 l / h and 150 l / h, preferably 120 l / h.

[0163] For illustration purposes, the reverse osmosis stage can be carried out on a reverse osmosis unit equipped with a SW 30 HR type membrane (Dow).

[0164] According to one embodiment, the reverse osmosis step is continued until a retentate of approximately 13 Bx corresponding to the osmosed ultrafiltration permeate is obtained.

[0165] 13 Bx or Brix corresponds to approximately 13% dry matter measured by the reduction index and expressed in degrees brix sucrose scale.

[0166] According to one embodiment, the obtained osmosis ultrafiltration permeate will undergo a drying step, for example by freeze-drying.

[0167] The fraction obtained of the osmosis ultrafiltration permeate type will hereinafter be referred to as the Ulva genus algae extract according to the invention or permeate or “fraction P”.

[0168] Thus, according to one embodiment, the method for obtaining an extract of algae of the genus Ulva comprises the following steps: a) Washing the algae to be treated b) Grinding the washed algae c) Extraction with water of the washed algae in a reduced volume of water d) Filtration of the pulps recovered after step c) then pressing of the pulps so as to obtain a pressing juice on the one hand and pressing pulps on the other hand e) Microfiltration of the pressing juice obtained in step d) so as to obtain a retentate on the one hand and a permeate on the other hand f) Ultrafiltration of the microfiltration permeate obtained in step e) so as to obtain a retentate on the one hand and a permeate on the other hand g) Reverse osmosis of the ultrafiltration permeate obtained in step f) h) Freeze-drying of the ultrafiltration permeate osmosed obtained in step g).

[0169] According to one embodiment, the algae extract obtained by the process and having anti-inflammatory properties has the following characteristics: a protein content of approximately 30%, an average content of total phenolic compounds of approximately 4% and approximately 10% of total sugars. Food supplement

[0170] According to one embodiment, the present invention also relates to a food supplement comprising an extract of algae of the genus Ulva according to the invention.

[0171] The algae extract of the genus Ulva is as defined above.

[0172] A "food supplement" means a food product intended to supplement a normal diet and which constitutes a concentrated source of nutrients or other substances having a nutritional or physiological effect.

[0173] According to one embodiment, the food supplement comprises, in addition to the algae extract according to the invention, at least one ingredient chosen from nutrients such as vitamins and minerals, plants and / or plant extracts, substances for nutritional or physiological purposes, adjuvants or food additives such as colorings, flavorings, preservatives.

[0174] The food supplement may also include all the pharmaceutically acceptable excipients as previously defined.

[0175] The food supplement according to the invention may be in any form compatible with oral absorption in one or more daily doses. It may be in particular in the form of capsules, tablets, lozenges or loose powder, the latter preferably being packaged in unit sachets.

[0176] It is particularly interesting to use the food supplement comprising an extract of the Ulva genus according to the invention in a food supplement, intended for overweight or obese patients, but also for overweight or obese pets.

[0177] According to one embodiment, the food supplement intended for humans is presented in the form of unit doses, for example in the form of capsules, capsules, tablets, lozenges or loose powder packaged in unit sachets, each unit dose comprising between 0.05 g and 2 g of algae extract according to the invention, preferably between 0.1 and 1 g, and more preferably between 0.2 and 0.8 g.

[0178] According to another embodiment, the food supplement intended for pets is presented in the form of unit doses, for example in the form of loose powder packaged in unit sachets, each unit dose comprising between 0.01 g and 2 g of algae extract according to the invention, preferably between 0.025 and 1.5 g, and more preferably between 0.03 and 1 g.

[0179] Surprisingly, the extract according to the invention shows significant activity at very low concentrations.

[0180] Without wishing to be bound by any theory, it is the inventors' opinion that concentration-dependent induction and inhibition mechanisms intervene or cancel each other out below a threshold concentration of the extract according to the invention given that this extract is a complex mixture of different molecules which can logically combine synergistic and / or antagonistic effects.

[0181] The present invention also relates to the use of a food supplement for its use in the therapeutic applications as described above. Food composition for humans

[0182] The algae extract according to the invention may be incorporated into any food composition intended for human consumption. Also, the algae extract according to the invention may be incorporated into drinks, meal replacements, biscuits, cereals, desserts, milk preparations, etc.

[0183] The present invention also relates to an algae extract in the form of a food composition for humans, characterized in that the food composition for humans is in a form chosen from drinks, meal replacements, biscuits, cereals, desserts, milk preparations.

[0184] It will be particularly interesting to use the food composition comprising an extract of the Ulva genus according to the invention in a food composition, intended for overweight or obese patients. Pet food composition

[0185] The present invention also relates to a food composition for pets comprising an extract of algae of the genus Ulva according to the invention.

[0186] These food compositions are not intended to be limited to a specific list of ingredients as these ingredients will depend on factors such as, for example, the desired nutritional balance for the specific type of pet, and the availability of the ingredients. In addition to the seaweed extract of the invention, the pet food composition may generally include vitamins, minerals and other additives such as flavorings, preservatives, emulsifiers and humectants. The nutritional balance, including the relative proportions of the seaweed extract of the invention and the other ingredients, is determined according to dietary standards known in the veterinary art. For example, the nutritional balance of a cat food composition is determined based on the known dietary needs of cats.

[0187] The present invention also relates to an algae extract in the form of a food composition for pets, characterized in that the food composition further comprises vitamins, minerals and other additives such as flavorings, preservatives, emulsifiers and humectants.

[0188] It will be particularly interesting to use the food composition comprising an extract of the Ulva genus according to the invention in a food composition intended for overweight or obese pets.

[0189] The present invention also relates to the use of a food composition for its use in therapeutic applications as described above. Examples

[0190] Example 1: Production and characterization of an algae extract of the genus Ulva

[0191] A. Production of the extract

[0192] This method corresponds to an adaptation of the extraction and fractionation process described in patent FR2998894B1 from the company Seprosys, allowing the purification of protein and polysaccharide fractions from green marine macroalgae.

[0193] The process comprises the following steps: a) Washing of the algae to be treated b) Crushing of the washed algae c) Diffusion of the washed algae into water in a reduced volume of water d) Filtration of the pulps recovered after step c) then pressing of the pulps so as to obtain a pressing juice on the one hand and pressing pulps on the other hand e) Microfiltration of the pressing juice obtained in step d) so as to obtain a retentate on the one hand and a permeate on the other hand f) Ultrafiltration of the microfiltration permeate obtained in step e) so as to obtain a retentate on the one hand and a permeate on the other hand g) Reverse osmosis of the ultrafiltration permeate obtained in step f) h) Freeze-drying of the osmosed ultrafiltration permeate obtained in step g).

[0194] 1 kg of dried seaweed is soaked in 30 L of raw water for 10 min at room temperature. The seaweed is then drained and pressed using a fabric cone to remove as much water as possible. The washed seaweed is then ground in 5 L of demineralized water at 65°C, using a knife mill, until particles less than 2 mm in diameter are obtained. The 5 L of ground material is transferred to a tank thermostatically controlled at 80°C containing 10 L of demineralized water at room temperature. The extraction is carried out under constant stirring using a paddle stirrer, at a rotation speed of 10 rpm for 2 h. The aqueous extract is then filtered using a vibrating sieve with a cut-off threshold of 50 μm (stainless steel cloth, NEGOFILTRE), in order to remove seaweed residues. The seaweed pulp is then drawn from the tank and pressed using a press (60 hl / h) to extract as much juice as possible.The juice from the press is then combined with the initial filtered aqueous extract and approximately 15 l of total extraction juice is recovered. then filtered via a plate filter type microfiltration unit equipped with two filters of 1.2 pm then 0.8 pm (3M, 40*40, pore size 15 HN and 50 HN). Approximately 15 I of permeate are recovered, then filtered again via an ultrafiltration unit equipped with a Kerasep KBW 15 kDa membrane (Novasep Process). Filtration is carried out at 80°C, 5 bars and at a circulation flow rate of 450 l / h, i.e. a circulation speed of 5 m / s. Filtration is continued until a retentate of approximately 4 Bx is obtained, which is then withdrawn from the unit. The permeate is recovered and then osmosed on a reverse osmosis unit equipped with a SW 30 HR membrane (Dow), allowing the fraction to be concentrated. Reverse osmosis is carried out at 30°C, between 20 and 30 bars and at a circulation flow rate of 120 l / h, i.e. a circulation speed of 1.3 m / s. Reverse osmosis is continued until a retentate of approximately 13 Bx is obtained, which is then withdrawn from the unit and then freeze-dried.This fraction obtained of the osmosis ultrafiltration permeate type will hereinafter be referred to as Ulva genus algae extract of the permeate type or “fraction P”.

[0195] Characterization of the extract F01961 a) Solubility

[0197] Several different solvents are used to evaluate the solubility of the permeate type Ulva algae extract (fraction P): water, acetonitrile (ACN), methanol, ethanol, acetone, hexane, water / ACN mixture (1:1, v / v), and methanol / acetone mixture (1:1, v / v). Different mass quantities of solid extract are mixed under vortex with the different solvents and then the absorbance of the solution or suspension obtained is measured at 600 nm using a FLUOstar Omega microplate spectrophotometer (BMG LABTECH). After subtraction of the blank obtained from each pure solvent, the solution is considered clear and the extract soluble if the absorbance obtained is less than 0.02.

[0198] The P fraction is soluble only in water and the water / ACN mixture (1:1, v / v) (Table 1), its solubility being maximum in water (1 g / l), and minimum in the water / ACN mixture (1:1, v / v).

[0199] [Table 1]

[0200] Table 1: Solubility of fraction P in water and water / acetonitrile mixture (1:1, v / v).

[0201] b) Biochemical characterization

[0202] Two solvents, water and water / ACN mixture (1:1, v / v), are used to prepare two solutions of fraction P at 20 mg / ml. These are then centrifuged at 15000 rpm for 2 min and then filtered through 0.22 µm to obtain a final solution containing only the soluble compounds. Due to the influence of the color of these two solutions on the absorbance measurement, they are then diluted 1 / 18 using the same solvent, before analysis.

[0203] All analyses are carried out in triplicate.

[0204] Protein levels

[0205] Protein content was assessed using three biochemical protocols, derived respectively from the Lowry (Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov; 193(1 ):265-75), Smith (Smith PK, Krohn RL, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, et al. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1 ):76-85) and Kjeldahl (Kirk PL. Kjeldahl Method for Total Nitrogen. Anal Chem. 1950 Feb 15;22(2):354-8) methods.

[0206] - Lowry method:

[0207] This involves the use of two reagents: Lowry's reagent, which consists of a mixture of 100 μl of 2% (m / v) potassium sodium tartrate solution, 100 μl of 1% (m / v) copper sulfate solution, and 10 ml of 2% (m / v) sodium carbonate solution in 0.1 M NaOH; and Folin's reagent, which is a 1 / 20 dilution of Folin-Ciocalteu's phenol reagent in ultrapure water.

[0208] In a 96-well microplate (NUNC 96 A / S, Denmark), 25 μl of the solution to be assayed is added to 75 μl of Lowry reagent before incubation for 20 min at room temperature and in the dark. Then 125 μl of Folin reagent is added to the reaction mixture. A second incubation of 30 min, at room temperature and in the dark, is applied before measuring the absorbance at 750 nm using a FLUOstar Omega microplate spectrophotometer (BMG LABTECH). A standard range of bovine serum albumin (BSA) from 50 to 600 pg.ml-1 is carried out simultaneously under the same conditions.

[0209] - Smith method, with bicinchoninic acid (BCA):

[0210] In a 96-well microplate (NUNC 96 A / S, Denmark), 200 μl of BCA reagent (bicinchoninic acid / 4% (m / v) copper sulfate (50:1, v / v)) and 10 μl of the solution to be measured are added. After incubation at 37°C in an oven for 30 min, the absorbance at 562 nm is measured using a FLUOstar Omega microplate spectrophotometer. (BMG LABTECH). A standard range of ASB ranging from 50 to 600 mg / l is carried out simultaneously under the same conditions.

[0211] - Kjeldahl method:

[0212] This method involves two successive steps, a first one of mineralization and a second one of distillation. For mineralization, 1 g of sample is added to a matra, with 2 catalyst tablets. A matra containing only the tablets is used as a blank and another matra containing a known quantity of urea as well as the 2 catalyst tablets is used as a control. 10 ml of 96% sulfuric acid are added to the matra. The solution is then heated for 2 h at approximately 400 ° C. After mineralization, distillation is carried out for 6 min after adding 50 ml of 30% (m / v) NaOH and 50 ml of ultrapure water. The liquid is then collected in a boric acid solution (4% (m / v), pH 4.65) and titrated with 1 M HCl.

[0213] Total sugar content

[0214] The determination of total sugars is carried out by the phenol / sulfuric acid mixture method introduced by Dubois (Dubois M, Gilles K, Hamilton JK, Rebers PA, Smith F. A Colorimetric Method for the Determination of Sugars. Nature. 1951 Jul;168(4265):167—167). In glass vials, 40 μl of the solution to be assayed is added to 40 μl of 5% (m / v) phenol and 200 μl of 96% sulfuric acid. The reaction mixture is incubated for 30 min at 90°C in the dark. The samples are then cooled in ice. The absorbance is then measured at 485 nm in a 96-well microplate using a FLUOstar Omega microplate spectrophotometer (BMG LABTECH). A standard range of rhamnose, a representative monomer, ranging from 0.025 to 0.6 mg / ml, is carried out simultaneously under the same conditions.

[0215] Total phenolic compound content

[0216] The determination of total phenolic compounds is carried out using the method of Singleton and Rossi (Singleton VL, Rossi JA. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am J Enol Vitic. 1965 Jan 1; 16(3): 144), with some modifications.

[0217] 10 μl of sample to be assayed and 790 μl of ultrapure water are introduced into centrifuge tubes. After mixing, 20 μl of 20% (m / v) Na2CO3 and then 50 μl of 2 N Folin-Ciocalteu reagent are added. The tubes are incubated at room temperature, in the dark, for 2 h. 200 μl of reaction medium are then transferred into a 96-well microplate and the absorbance at 730 nm is measured using a FLUOstar Omega microplate spectrophotometer (BMG LABTECH). A gallic acid standard range, composed reference phenolic, ranging from 10 to 150 pg / ml, is carried out simultaneously under the same conditions.

[0218] The biochemical characterization of the P fraction shows that this fraction has a significant protein content, ranging from 28.6% to 31.6% depending on the dosage method applied and the nature of the solvent used for its solubilization. These very close results make it possible to average this total protein content at approximately 30%. In the same way, the total phenolic compounds are quantified at very similar values, regardless of the solvent used, giving an average content of approximately 4%. The results obtained in terms of total sugar content are, however, very different, the content measured in the water / ACN mixture (1:1, v / v) being much lower (3.3%) than that measured in water (10.0%). This is consistent, given the lower solubility of oligo- and / or polysaccharides resuspended in a mixture composed of water and ACN, which is a less polar solvent, whereas they were initially extracted in pure water.In conclusion, the extract contains 10% total sugars.

[0219] [Table 2]

[0220] Example 2: Production and characterization of an algae extract of the genus Ulva

[0221] To evaluate the in vitro anti-inflammatory effect of the Ulva algae extract of permeate type (fraction P), the latter was tested on a murine macrophage cell line Raw 264.7 provided by ATCC Cell ("American Type Culture Collection"): code ATCC-TIB-71. These are immortalized mouse macrophages used as reference biological study models in metabolic diseases, to measure anti / pro-inflammatory activity (Jones E, Adcock IM, Ahmed BY, Punchard NA. Modulation of LPS stimulated NF-kappaB mediated Nitric Oxide production by PKCc and JAK2 in RAW macrophages. Journal of Inflammation. 2007 Nov 24;4(1):23).

[0222] For all of these evaluations, a one-way ANOVA statistical test was performed using Origin 6 software (OriginLab Corporation).

[0223] A. Effect of an Ulva algae extract (permeate) on the cell viability of Raw 264.7 macrophages in culture:

[0224] Cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) (PAN Biotech, Dutscher) supplemented with 10% (v / v) fetal bovine serum (PAN Biotech, Dutscher) and 1% (v / v) antibiotic solution (10,000 U / ml penicillin, 10 mg / ml streptomycin) (PAN Biotech, Dutscher). This medium, hereinafter referred to as "complete medium", was sterilized by 0.22 µm filtration. Cells were cultured in a temperature-controlled incubator at 37°C, with 5% CO2, under a humid atmosphere, in 75 cm ventilated flasks. 2(BD Biosciences), and maintained by mechanical removal with a scraper (Sarstedt). The culture medium is renewed every 2 or 3 days. All manipulations are carried out under a class 2 microbiological safety cabinet. For experiments, the cells are used between n ème and I6 ème passages.

[0225] The effect of Ulva algae extract of permeate type (fraction P) on cell viability is determined via the enzymatic conversion of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983 Dec 16;65(1-2):55-63). This assay is based on the reduction of tetrazolium sei by mitochondrial succinate dehydrogenase to Formazan crystals. Cells are seeded in sterile 96-well microplates (BD Biosciences), at a concentration of 5x104 cells / ml in a volume of 100 µl of complete medium. After 24 h of incubation, the complete medium was replaced with 100 μl of complete medium containing the P fraction at different concentrations (between 0.01 and 500 pg / ml), or 1 μM dexamethasone (anti-inflammatory drug compound, used as a positive control. A negative control containing only complete medium was also used. After 24 h of exposure, 25 μl of a 5 mg / ml MTT solution in PBS was added to each well, which were then incubated for 4 h in the culture incubator. The medium was then removed and 200 μl of DMSO (dimethyl sulfoxide) was added to each well to solubilize the formed Formazan crystals. After 10 min of incubation at room temperature, the absorbance at 550 nm was measured using a Fluostar Omega microplate reader (BMG LABTECH).

[0226] Cell viability is determined according to the following formula:

[0227] [Math.1]

[0228] Figure 1A shows the impact of the P fraction on the viability of Raw 264.7 macrophages, compared to control conditions, after 24 hours of exposure. Figure 1B shows the same experiment in the presence of Salmonella lipopolysaccharide (LPS) at 1 pg / ml (LPS 0111, reference L2630, Merck), after 6 hours of pre-incubation with the P fraction at different concentrations. In both cases, the effect of a reference anti-inflammatory drug used as a positive control, dexamethasone (1 pM), is also compared. The results obtained demonstrate that neither the P fraction, whatever its concentration, nor dexamethasone at 1 pM have a significant effect on the cell viability of Raw 264.7 macrophages, whether they are cultured in the presence of LPS or not.

[0229] B, Effect of an Ulva algae extract (permeate) on nitric oxide (NO) secretion by Raw 264.7 macrophages in culture, under basal or proinflammatory conditions

[0230] Nitric oxide (NO) is a central mediator of the inflammatory process, regulating the activity, growth, and death of many types of immune and inflammatory cells (Grisham MB, Jourd'Heuil D, Wink DA. Nitric oxide. I. Physiological chemistry of nitric oxide and its metabolites: implications in inflammation. Am J Physiol. 1999 Feb;276(2):G315-321 , Lee M, Rey K, Besler K, Wang C, Choy J. Immunobiology of Nitric Oxide and Regulation of Inducible Nitric Oxide Synthase. Results Probl Cell Differ. 2017;62:181-207). NO assay can be performed from culture supernatants of RAW 264.7 macrophages according to the protocol described by Li and Shah (Li S, Shah NP. Characterization, Anti-Inflammatory and Antiproliferative Activities of Natural and Sulfonated Exo-Polysaccharides from Streptococcus thermophilus ASCC 1275. Journal of Food Science.2016;81(5):M1167-76), which involves the reaction of Griess reagent with NO, resulting in the formation of a stable chromophore absorbing at 540 nm.

[0231] The cells and culture conditions are the same as those described in Example 2, Part A. The cells are seeded in sterile 96-well microplates (BD Biosciences), at a rate of 1.5.10 6cells / ml, in complete DMEM medium. After 24 h of incubation, the initial medium is replaced by 100 μl of complete medium containing the P fraction at different concentrations (between 0.01 and 100 pg / ml), or dexamethasone at 1 pM (reference anti-inflammatory compound used as a positive control), before another 6 h of incubation. A negative control containing only complete medium is also carried out. To evaluate the anti-inflammatory effects of the permeate-type Ulva algae extract (P fraction), the same experiment is carried out in parallel, this time with 6 h of pretreatment in the presence of permeate at various concentrations. concentrations, or dexamethasone at 1 pM, followed by incubation in the presence of salmonella lipopolysaccharide (LPS) at 1 pg / ml (LPS 0111, reference L2630, Merck) for 18 h.

[0232] The Griess Reagent Kit for Nitrite Determination (G-7921, Invitrogen™, Thermo Fischer Scientific) is used to measure the NO concentration in cell culture supernatants, according to the protocol provided by the supplier. 150 μl of cell supernatant is collected for determination using an equivalent volume of Griess reagent. After incubation at room temperature for 30 min, the absorbance is measured at 540 nm using a Fluostar Omega microplate reader (BMG LABTECH).

[0233] Figure 2 shows the effect of the P fraction on NO secretion in the culture medium under basal (A) or inflammatory (B) conditions. An increase in NO secretion compared to control conditions is observed in the presence of LPS (positive control), as well as for the two highest concentrations of P fraction (10 and 100 pg / ml) (Figure 2A).

[0234] In contrast, the P fraction induced a very significant decrease in NO production in the presence of LPS (Figure 2B), with the NO concentration in the culture supernatant being halved at a concentration of 1 pg / ml and by approximately 1.5 at concentrations of 0.01 and 0.1 g / ml, compared to inflammatory control conditions involving LPS alone. These results indicate that the P fraction is very strongly anti-inflammatory because it induces a reduction in NO secretion close to that induced by dexamethasone, at very low concentrations.

[0235] C. Effect of an Ulva algae extract (permeate) on the expression of key proteins involved in signaling pathways related to LPS-induced inflammation, by cultured Raw 264.7 macrophages

[0236] The evaluation of the modulation by the permeate-type Ulva algae extract (fraction P) of protein signaling pathways involved in inflammation, complements and clarifies that of nitric oxide secretion by Raw 264.7 macrophages in culture. This is carried out by Western Blot analysis of protein biomarkers involved in the inflammatory process: NLRP-3 and iNOS. NLRP-3, also called inflammasome, belongs to a class of cytoplasmic protein complexes (NLRP) that modulate the activation of inflammatory mediators. NLRP-3 plays a pivotal role among the NLRP class, and is activated after stimulation by microbial and non-microbial factors such as bacterial toxins, micro-particles or LPS. It promotes the expression, maturation and secretion of a multitude of pro-inflammatory cytokines (Wang Z, Zhang S, Xiao Y, Zhang W, Wu S, Qin T, et al. NLRP3 Inflammasome and Inflammatory Diseases. Oxid Med Cell Longev. 2020 Feb 17;2020:4063562). Inducible nitric oxide synthase (iNOS) catalyzes the synthesis of nitric oxide (NO), which is a key mediator of inflammation. LPS, interferon gamma (IFN-γ), TNF-α, and IL-1β can induce iNOS expression. (Lee M, Rey K, Besler K, Wang C, Choy J. Immunobiology of Nitric Oxide and Regulation of Inducible Nitric Oxide Synthase. Results Probl Cell Differ. 2017;62:181-207).

[0237] The Western Blot method is a semi-quantitative analysis of protein expression in cells. The cells and culture conditions are the same as those described in Example 2, Part A. Raw 264.7 cells are seeded in sterile 6-well microplates (BD Biosciences), at 1x10 6cells / ml, in 1 ml of complete DMEM medium. After 24 h of incubation, the initial medium is replaced by 1 ml of complete medium containing the P fraction at different concentrations or dexamethasone at 1 pM (reference anti-inflammatory compound used as a positive control), before another 6 h of incubation. A negative control containing only complete medium is also prepared. LPS at 1 pg / ml is then added to all wells, except the one corresponding to the control conditions, then incubation is continued for 18 h. The cell supernatant is removed and then a cell lysate is prepared in the lysis buffer (RIPA, Thermo Fisher Scientific) supplemented with a phosphatase inhibitor (Cell Signaling). The cell lysate is centrifuged for 5 min at 10,000 g, at 4°C. The protein concentration in the supernatant is then obtained by applying the Smith method, to bicinchoninic acid (BCA), according to the protocol given in example 1.An equivalent amount of protein for each sample (25 pg) is mixed with loading buffer (Bio-Rad) and p-mercaptoethanol (Merck) to reach a final volume of 50 μl. The samples are then heated at 95 °C for 5 min and then 10 μl are deposited, after returning to room temperature, in gels with different concentrations of polyacrylamide depending on the molecular weight of the protein studied. Size markers are also added to the gel (Seeblue® plus 2, Thermo Fisher Scientific). Migration is carried out at 150 V for 45 min in migration buffer. The proteins are then transferred from the gel to a nitrocellulose membrane (Bio-Rad), at 100 V for 7 min in transfer buffer using the semi-liquid transfer method (Bio-Rad). The membrane is then blocked for 1.5 hours at room temperature, using a blocking solution containing 5% bovine serum albumin (BSA) in TBS-Tween 20 (Merck).The membrane is incubated in the presence of the primary antibody, overnight at 4°C, then in the presence of the secondary antibody coupled to horseradish peroxidase, for 1.5 hours at room temperature (15).

[0238] [Table 3]

[0239] Table 3: Primary and secondary antibodies used for Western blot analysis of signaling pathways involved in LPS-induced inflammation.

[0240] The development solution (SuperSignal™ West Femto Maximum Sensitivity Substrate, Thermo Fischer Scientific) is applied to the membrane for 1 min, then acquisition is performed by a Chemidoc system (Bio-Rad). Analysis of band densities is performed by ImageLab software (Bio-Rad) and the results are expressed relative to the amount of O-actin protein present in the samples, chosen as a housekeeping protein.

[0241] This Western blot analysis proves that the P fraction induces a drastic reduction in the expression of NLRP3 and iNOS proteins, involved in the inflammatory process in Raw 264.7 macrophages in culture (Figure 3). This effect is obtained in a dose-dependent manner for NLRP-3, the P fraction inhibiting its expression from the concentration of 1 pg / ml.

[0242] Conversely, iNOS expression is reduced only at low concentrations (0.1 and 1 pg / ml), with the P fraction having no significant activity at 10 pg / ml. This reversed dose-response effect may have two main causes. Either the intervention of concentration-dependent induction and inhibition mechanisms, which intervene or cancel each other out below a threshold concentration, the P fraction being in fact a very complex mixture of different molecules that can logically associate synergistic and / or antagonistic effects. Or the possible involvement of a hormesis phenomenon, a toxicology term designating a two-phase biological response to a compound, characterized by stimulation at low doses associated with a beneficial effect, and by inhibition at high doses associated with a toxic effect.The second hypothesis is very unlikely, given that the P fraction is not cytotoxic at the concentrations studied, as proven by the commonly used MTT cell viability test, presented in example 2 (part A), in the presence or absence of LPS.

[0243] Taken together, these results fully corroborate the anti-inflammatory effect demonstrated by the P fraction against NO production by Raw 264.7 macrophages in culture, presented in Example 1 (Part B).

[0244] D. Effect of an Ulva algae extract (permeate) on the secretion of pro-inflammatory cytokines by Raw 264.7 macrophages in culture, in the presence of LPS:

[0245] The evaluation of the modulation by the P fraction of the secretion of proinflammatory cytokines in the extracellular medium also complements and clarifies that of the secretion of nitric oxide by Raw 264.7 macrophages in culture, as well as the protein signaling pathways involved in inflammation. This is carried out by ELISA method, for the quantitative analysis of two pro-inflammatory cytokines secreted by cells and present in the extracellular medium: TNF-g and IL-6.

[0246] The cells and culture conditions are the same as those described in Example 2, Part A. Raw 264.7 cells are seeded in sterile 6-well microplates (BD Biosciences), at 1x10 6cells / ml, in 1 ml of complete DMEM medium. After 24 h of incubation, the initial medium is replaced by 1 ml of complete medium containing the P fraction at different concentrations (between 0.01 and 100 pg / ml), or dexamethasone at 1 pM (reference anti-inflammatory compound used as a positive control) before another 6 h of incubation. A negative control containing only complete medium is also prepared. LPS at 1 pg / ml is then added to all wells, except the one corresponding to the control conditions, and then incubation is continued for 18 h. The culture supernatant is then removed and frozen at -80°C. The concentrations of TNF-β and IL-6 in this supernatant are then determined according to the method indicated by the kit supplier (Peprotech), involving two final absorbance measurements, one for evaluation, at 405 nm, and the other for correction, at 650 nm, carried out using a Fluostar Omega microplate reader (BMG LABTECH).

[0247] The secretion of both pro-inflammatory cytokines is significantly increased in the presence of LPS, compared to control conditions (Figure 4). The P fraction also exhibits this pro-inflammatory effect at high concentrations: from 1 pg / ml for TNF-β secretion and 10 pg / ml for IL-6 secretion. Below these threshold concentrations, no significant difference is observed compared to control conditions.

[0248] After 6 h of pre-incubation with the P fraction and in the presence of LPS, the secretion of TNF-o and IL-6 is drastically reduced, reaching a level similar to that obtained in the presence of dexamethasone, the reference anti-inflammatory compound, even at the very low concentration of 0.1 pg / ml.

[0249] Taken together, these results demonstrate the anti-inflammatory effect of the P fraction with respect to NO production, expression of key proteins involved in signaling cascades linked to the inflammatory process (NLRP-3 and iNOS), and the production of pro-inflammatory cytokines.

[0250] Example 3: Demonstration of in vivo efficacy of an Ulva algae extract (permeate) on a model of diet-induced behavioral alterations

[0251] Adult male C57BI / 6J mice, supplied by Janvier and housed in the NutriNeuro laboratory animal facility, were fed either a nutritionally balanced diet (A04 diet) or a high-calorie diet enriched in fat and sugar (HFD diet) for 12 weeks to induce weight gain associated with metabolic syndrome and behavioral alterations according to a protocol classically described in the literature and used in the NutriNeuro laboratory (Zemdegs J, Quesseveur G, Jarriault D, Pénicaud L, Fioramonti X, Guiard BP. High-fat diet-induced metabolic disorders impairs 5-HT function and anxiety-like behavior in mice. Br J Pharmacol. 2016 Jul; 173(13):2095—110, Zemdegs J, Martin H, Pintana H, Bullich S, Manta S, Marqués MA, et al. Metformin Promotes Anxiolytic and Antidepressant-Like Responses in Insulin-Resistant Mice by Decreasing Circulating Branched-Chain Amino Acids. J Neurosci. 2019 Jul 24;39(30):5935-48).

[0252] To evaluate the effect of the permeate-type Ulva algae extract (fraction P) on metabolic and behavioral alterations, a 1 mg / ml P fraction supplement was provided to the animals via drinking water for 4 weeks following 12 weeks of HFD diet. At the end of this supplementation, metabolic (weight, fat mass and lean mass) and behavioral (anxiety-like behavior, memory capacity) parameters were evaluated.

[0253] For all these evaluations, a one-way ANOVA statistical test associated with a post-hoc Tuckey test was carried out using GraphPad Prism 7 software (Graph Pad Software).

[0254] A. Impact of Ulva algae extract supplementation (permeate) on metabolic parameters

[0255] After supplementation, the animals were weighed to assess the impact of supplementation on weight gain. As expected, mice fed a HFD diet had a significantly higher weight than mice fed the control diet (AO4) (F (2, 22) = 12.34, p<0.001) (Figure 5). In addition, P fraction supplementation induced a statistically significant weight loss in supplemented HFD mice compared to mice not receiving the supplement (p<0.01). The weight of HFD mice supplemented with P fraction is not statistically different from that of control mice.

[0256] Analysis using a 3T MRI device with spin echo sequences allowed the quantification of lean mass and fat mass of mice. As expected, mice fed with a HFD diet have a significantly lower amount of lean mass than control mice (F (2, 22) = 35.11, p < 0.001), with no effect of supplementation. However, unsupplemented HFD mice have a significantly higher amount of fat mass than control mice (F (2, 22) = 36.59, p < 0.001). But supplementation with P fraction significantly reduces the amounts of fat mass in mice fed with a HFD diet (F (2, 22) = 36.59, p < 0.01).

[0257] Supplementation with P fraction thus protects against fat mass gain induced by the HFD diet.

[0258] B. Impact of supplementation with Ulva algae extract (permeate) on the >

[0259] a) Assessment of anxiety disorders

[0260] The animals' anxiety-like behavior was assessed in the elevated plus maze test. Two opposite arms were without walls (open) and the other two had 10 cm high walls (closed). In the center was a platform open to all four arms. Mice were placed on the central platform at the beginning of the test and were free to explore the maze for 5 min. The time spent in the different areas was measured by the SMART video-tracking system (Bioseb). This device allows the determination of the anxiety state of the animals, which will spontaneously avoid the naturally anxiety-inducing open arms and prefer the closed arms. It has been shown using this device that pharmacological treatments with anxiolytics were effective in counteracting the natural aversion caused by open arms.

[0261] As expected, HFD-fed and unsupplemented animals showed a significant decrease in time spent in the open arms of the elevated plus maze (F(2, 20) = 7.376, p<0.05) and thus exhibited anxiety-like disorders (Figure 5). Fraction P supplementation restored the animals' exploration time in the open arms (F(2, 20) = 7.376, p<0.01). Fraction P supplementation thus protected against the onset of anxiety-like disorders induced by the HFD diet.

[0262] Evaluation of memory impairments

[0263] To assess the chronic effects of treatment on memory, mice are tested in the Morris water maze.

[0264] The water maze test developed by Morris (Morris R. Developments of a water-maze procedure for studying spatial learning in the rat. J Neurosci Methods. 1984 May;11(1):47-60.), allows the measurement of learning and spatial memory capacities in animals. In this experimental protocol, this test specifically involves the hippocampus-dependent spatial reference memory. The animal is placed in a swimming pool and must learn to locate a refuge platform using visual cues distributed on the walls of the room.

[0265] To carry out this test, we have a circular swimming pool (150 cm in diameter), filled with water (35 cm deep, temperature: 22±2°C), made opaque by the addition of white paint. The platform (10 cm in diameter) is submerged 5 cm below the water surface and is located 30 cm from the edge of the pool. The chosen protocol takes place in 4 stages (figure 7): 1 - Habituation: allows the mice to become familiar with swimming and climbing onto a platform. This stage takes place in a circular basin (65 cm in diameter) filled with clear water. 2-cued learning: allows the presence of motor or visual deficits to be assessed and accentuates familiarization. The animals must find the submerged platform indicated by a clue. 3-reference spatial learning: animals must find a submerged and invisible platform using distal visual cues. 4- the restitution test: allows spatial memory to be assessed. The mice are placed in the pool in the absence of a platform and must search for 1 minute for the place where the platform was initially positioned.

[0266] Mouse movements are recorded by a video-tracking system (SMART), which allows for each trial to obtain the latency to reach the platform as well as the total distance traveled in the pool. The swimming speed of the mice is deduced from these values. The time spent in the different quadrants as well as the distance traveled and the number of entries in the target quadrant are calculated.

[0267] In our protocol, control animals fed a balanced diet do not show cognitive impairment and significantly discriminate the target quadrant compared to random exploration (p<0.001). As expected, animals on a HFD diet are not able to recognize the target quadrant and therefore show impairments in their long-term spatial memory (Figure 8). Interestingly, the Supplementation with fraction P restores the cognitive abilities of the animals since they statistically significantly differentiate the target quadrant from chance (p<0.05). There is no difference in learning between the groups.

Claims

Claims

1. Extract of algae of the genus Ulva characterized in that: - the protein content is between 5 and 60% by mass, preferably between 15 and 50% by mass, preferably between 25 and 35% by mass, relative to the total dry mass of the extract, - the content of total phenolic compounds is between 0.5 and 40% by mass, preferably between 1 and 20% by mass, preferably between 1 and 10% by mass, relative to the total dry mass of the extract, - the total sugar content is between 0.5 and 80% by mass, preferably between 1 and 50% by mass, preferably between 5 and 15% by mass, relative to the total dry mass of the extract, for its use in the treatment of disorders induced by metabolic syndrome.

2. Algae extract for its use according to claim 1 characterized in that the disorders induced by a metabolic syndrome are chosen from inflammation induced by a metabolic syndrome and cognitive and / or emotional disorders induced by a metabolic syndrome.

3. Algae extract for use according to claim 1 or 2, characterized in that the inflammation or cognitive and / or emotional disorders are induced by obesity or overweight.

4. Algae extract for its use according to claim 2 or 3 characterized in that the cognitive and emotional disorders are chosen from hippocampo-dependent long-term memory disorders and anxiety-type disorders.

5. Algae extract for use according to any one of claims 1 to 4, characterized in that the protein content is between 28 and 32% by mass relative to the total dry mass of the extract.

6. Algae extract for use according to any one of claims 1 to 4, characterized in that the content of total phenolic compounds is between 2 and 6% by mass relative to the total dry mass of the extract.

7. Algae extract for use according to any one of claims 1 to 4, characterized in that the total sugar content is between 5 and 15% by mass relative to the total dry mass of the extract.

8. Algae extract for use according to any one of claims 1 to 7, characterized in that the alga of the genus Ulva is chosen from Ulva lactuca, Ulva rigida, Ulva conglobata, Ulva ohnoi, Ulva reticulata, Ulva prolifera, Ulva flexuosa and Ulva intestinalis.

9. Algae extract for use according to any one of claims 1 to 7, characterized in that the alga of the genus Ulva is chosen from Ulva lactuca and Ulva rigida.

10. Algae extract for use according to any one of the preceding claims, characterized in that the alga extract is present in the form of a pharmaceutical composition, a veterinary composition, a food supplement or a food composition for humans or pets.

11. Algae extract for its use according to claim 10 characterized in that the pharmaceutical or veterinary composition further comprises a pharmaceutically acceptable excipient.

12. Algae extract for its use according to claim 10 characterized in that the food supplement further comprises at least one ingredient chosen from nutrients such as vitamins and minerals, plants and / or plant extracts, substances for nutritional or physiological purposes, adjuvants or food additives such as colorings, flavorings, preservatives, and a pharmaceutically acceptable excipient.

13. Algae extract for its use according to claim 10 characterized in that the food composition for humans is in a form chosen from drinks, meal replacements, biscuits, cereals, desserts, milk preparations.

14. Seaweed extract for use according to claim 10 characterized in that the food composition further comprises vitamins, minerals and other additives such as flavorings, preservatives, emulsifiers and humectants.

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