WO2019243887A1

WO2019243887A1 - Method for restoration of the natural capacity of the body to end inflammation by personalization of the food supplementation and objectification of its effect

Info

Publication number: WO2019243887A1
Application number: PCT/IB2019/000649
Authority: WO
Inventors: Marc DUBOURDEAU
Original assignee: Ambiotis
Priority date: 2018-06-22
Filing date: 2019-06-24
Publication date: 2019-12-26

Abstract

The present invention is drawn to a method for restoration of the natural capacity of the body of an individual to end inflammation by personalization of the food supplementation and objectification of its effect through a metabolo-lipodomic based blood test of said individual. The present invention also relates to dietary supplement and kits for the implementation of the method.

Description

Method for restoration of the natural capacity of the body to end inflammation by personalization of the food supplementation and objectification of its effect

Inflammation is a silent killer that participates in many disorders that impaired quality of life of many persons. It shortens lifespan by amplifying some diseases such as diabetes, cancer or neurodegenerative disease.

Until now it was thought that stopping inflammation was a passive process due to the catabasis of pro-inflammatory molecules. So far, companies sell dietary supplements that are based on anti-inflammatory compounds (like curcumin, resveratrol, harpagophytum...).

Indeed, the paradigm currently employed in the treatment of inflammatory conditions is to inhibit pro-inflammatory components. But the intake of anti-inflammatory compounds that are not evaluated on their pro-resolving capacities can be toxic for resolution and lead to adverse effects or even delay the end of inflammation.

In France the INCA 3 study (ANSES June 2017) revealed that the French diet lacks fats. This was likely due to the former advice to reduce fat intake, and it now appears that fat intake is not sufficient to provide enough omega-3 fatty acids especially EPA and DHA. EPA and DHA are involved in the inflammation regulation, and it has been demonstrated that their intake is important to stay in good health and to prevent the development of a chronic inflammation. There is thus a need for dietary supplements to complement diet in EPA and DHA.

It has been identified that novel molecules produced from long chain omega-3 and -6 fatty acids promote the termination of inflammation. These mediators are termed specialized pro-resolving mediators (SPMs). Pr Charles N. Serhan at Harvard Medical School initially discovered them. Over the last 15 years, many academic groups worldwide have largely demonstrated their beneficial biological actions. It has been demonstrated that many inflammatory conditions were associated with a defect in resolution of the inflammation.

For example, the document WO2018098244A1 (Sheran C. et al.) can be cited which describes a method of providing a metabolo-lipidomic profile and SPM signature on the progress of the innate host defense response following blood clotting or coagulation and wherein pro -thrombotic and pro-inflammatory mediators and SPMs are measured in a patient's blood sample in order to make medical or therapeutic decisions.

W02013170006A2 (Bannenberg G. et al.) can be also cited which describes a method of treating an inflammatory condition comprising the administration of an effective amount of oils having anti-inflammatory or resolution-stimulating activity and characterized in that they contain or are enriched with SPMs or SPM precursor originates from oil obtained from organisms containing long chain omega-3 polyunsaturated fatty acids. Acting positively on the biological pathways to produce SPMs is now considered as a novel strategy to fight chronic inflammatory conditions but also what we called low-grade or “silent” inflammation, often identified in age-related disorders. This branch is called the pharmacology of resolution and aims at restoring the resolution.

Few treatment or food supplementation deal with the resolution of inflammation. As instance, the Navamepent drug has been developed to improve resolution and has shown positive effect on dry eye syndrome in a phase 2 clinical study. Moreover, some companies use food supplementation that are enriched with SPM to boost resolution. However none claims for personalization of food supplementation and objectification of its effects on the resolution of inflammation.

Recently there is a trend to move from Evidence-Based Medicine to the 4P medicine, a new approach which is Predictive, Preventive, Personalised and Participatory because people are more and more aware that a drug or a dietary supplement does not fit to everyone (different health status, different microbiota, different lifestyles).

The evolution of our post-industrial society, lifestyle (sedentary, diet, overmedication...), the environmental causes (pollution...) and aging let to discover new disorders that are partly issued from an imbalance in the inflammatory response. Today, consumers ask for product claims to be proven, for products with proven benefits. With the development of « civilization diseases », people are more and more proactive about their health and want personalized solutions with proven effects that is lacking in the dietary supplements market, there is a need to provide with food dietary supplement, with proven efficacy on promotion of the resolution of inflammation, and their use to fight chronic inflammatory conditions but also what we called low-grade or“silent” inflammation, often identified in age-related disorders. The inventor propose with the present invention a solution to enhance the inflammation resolution capacity of the organism without adverse effects, without negative impact on the environment and with benefits for the society⁴ s Health management.

This is the object of the present invention that consists in a personalized nutritional approach objectified by the evaluation of plasma or serum biomarkers of inflammation resolution.

It has been discovered that blocking of pro -inflammatory molecules is not sufficient and that the body has to produce new molecules that are called SPMs to actively end inflammation. In other terms, without secretion of SPMs inflammation is not able to correctly stop and a defect of resolution is associated with all uncontrolled inflammations.

These dietary supplements will participate in the prevention of uncontrolled inflammations and in the relief of the minor daily health troubles.

In a first aspect, the present invention is directed to a method for determining or to predict whether resolution in an individual, suffering of an uncontrolled inflammation, can be improved or if said uncontrolled inflammation is likely to last longer or increase, preferably whether the resolution of inflammation can be improved by the ingestion of dietary supplement containing SPMs or selected compounds that have been tested to improve SPM production during uncontrolled inflammation, said method comprises the following steps :

a) from body fluids sample of said individual, preferably blood sample, evaluating the plasma or serum biomarkers of inflammation resolution, said plasma or serum biomarkers comprising at least a set of pro-inflammatory biomarkers and a set of pro-resolving biomarkers, b) determining the resolution of inflammation index of said individual thanks to comparison of the balance between pro-inflammatory and pro-resolving biomarkers, and

c) determine whether the inflammation will stop or if it is likely to last longer or increase, wherein:

- pro-resolving mediators are produced in higher amount versus pro-inflammatory mediators that are produced in lower amount is significant of inflammation which can be stopped, preferably is significant inflammation which can be stopped by the ingestion of dietary supplement containing SPMs;

- no or less production of pro-resolving mediators versus same or higher level of pro- inflammatory mediators is significant of inflammation which is likely to last longer or increase.

In step c), in a preferred embodiment, by higher amount for pro-resolving mediators, versus pro-inflammatory mediators, it is intended to designate at least a resolution of inflammation index of at least 2, prefably 2.5 , 3 , 3.5 before SPM and theis precursors supplemention by ingestion of the dietary supplement.

In another preferred embodiment, by higher amount for pro-resolving mediators, versus pro-inflammatory mediators, it is intended to designate at least a resolution of inflammation index of at least 6, preferably 8, 9, 15, 25 after SPM and theis precursors supplemention by ingestion of the dietary supplement.

In a preferred embodiment, the present invention is directed to a method for determining or to predict whether resolution in an individual, suffering of an uncontrolled inflammation, can be improved or if said uncontrolled inflammation is likely to last longer or increase, preferably whether the resolution of inflammation can be improved by the ingestion of dietary supplement containing SPMs or selected compounds that have been tested to improve SPM production during uncontrolled inflammation, said method comprises the following steps :

- pro-resolving mediators are produced in higher amount between two measurements versus pro-inflammatory mediators that are produced in lower amount between two measurements is significant of inflammation which can be stopped, preferably is significant inflammation which can be stopped by the ingestion of dietary supplement containing SPMs;

- no or less production of pro-resolving mediators versus same or higher level of pro- inflammatory mediators is significant of inflammation which is likely to last longer or increase; - each measurement is separated by a minimum of at least 3 weeks , preferably 1 month, more preferably 1 ,5 month, and preferably,

- wherein, during these two separated measurements, a first dietary supplement containing SPMs and/or their precursors issued from omega 3 (EPA, DHA) rich oil, is taken by said individual.

In a more preferred embodiment, said first dietary supplement containing SPMs or their precursors issued from omega 3 (EPA, DHA) rich oil, taken by said individual during the period of the two measurements includes at least a quantity od about 250 mg to 1000 mg of said first dietary supplement containing SPMs or their precursors, preferably 350 to 750 mg, more preferably 400 to 600 mg, 500 mg being the most preferred, said first dietary supplement comprising at least level of 18-HEPE, 14-HDOHE and 17-HDOHE egal or comprised between the intermediate and the high value as indicated in the following Table 1. Table 1 : Example of high level of SPMs or their precursors contained in omega 3 (EPA,

DHA) rich oil, compared with intermediate level.

Thus, according to the present invention, the first measurement allows to obtain the basis level of the resolution of inflammation index and the second its evolution, preferably following treatment with supplement dietary containing SPMs or their precursors issued from omega 3 (EPA, DHA) rich oil, by calculating whether higher, no or less production of pro-resolving mediators have been produced by the individual. Uncontrolled inflammation or inflammation which can be correctly stopped with secretion of SPMs can be defined herein as following. As disclosed by Chen et al. (Onconflammation is the immune system's response to harmful stimuli, such as pathogens, damaged cells, toxic compounds, or irradiation [1, and acts by removing injurious stimuli and initiating the healing process . Inflammation is therefore a defense mechanism that is vital to health. Usually, during acute inflammatory responses, cellular and molecular events and- interactions efficiently minimize impending injury or infection. This mitigation process contributes to restoration of tissue homeostasis and resolution of the acute inflammation. However, uncontrolled acute inflammation may become chronic, contributing to a variety of chronic inflammatory diseases. Among other reasons for chronic inflammation pro- inflammatory diets and ageing process (inflammaging) highly participate in uncontrolled inflammation.

By pro-resolving mediators, it is intended herein to designate the compound selected from the group consisting of:

- precursors of pro-resolving mediators 17HDoHE (17-Hydroxy-4,7,10,13,15,19- docosahexaenoic acid), MHDoHE (14-Hydroxy-4Z,7Z,10Z,12E,16Z,19Z-docosahexaenoic acid), 18HEPE (18-hydroxy-5Z,8Z,l lZ,l4Z,16E-eicosapentaenoic acid), 12HETE, 15HETE (12-, 15-Hydroxyeicosatetraenoic acid); and

- SPMs selected from the group consisting of: LXA4 (lipoxin A4), LXB4 (lipoxin B4),

RVE1, RVE2, RVE3 (resolvins El, E2, E3), RVD1, RVD2, RVD3, RVD4, RYD5, RVD6 (Resolvins Dl, D2, D3, D4, D5, D6), PD1 (protectin Dl), MaRl and/or MaR2 (maresin 1 and/or 2). By pro-inflammatory mediators, it is intended herein to designate the compound selected from the group consisting of:

- pro inflammatory PGE2 (prostaglandin E2), LTB4 (leukotriene B4), TXB2 (thromboxane B2) biomarkers; and

- the precursor 5HETE (5-Hydroxyeicosatetraenoic acid).

In another aspect, the present invention is directed to a method to increase the resolution of inflammation of an individual, comprising the following steps of:

a) from body fluids sample of said individual, preferably blood sample, evaluating the plasma or serum biomarkers of inflammation resolution, said plasma or serum biomarkers comprising at least a set of pro-inflammatory biomarkers and a set of pro-resolving biomarkers, b) determine the resolution of inflammation index of said individual thanks to comparison of the balance between pro-inflammatory and pro-resolving biomarkers c) define the composition of a personalized dietary supplement necessary to increase said resolution of inflammation index of said individual, preferably based on the value of the resolution of inflammation index obtained in step B) and, more preferably, further on the nature and the concentration of the biomarkers identified in step a), and

d) ingestion by said individual of said personalized dietary supplement.

In a preferred embodiment, the present invention is related to a method to increase the resolution of inflammation of an individual, comprising the following steps of:

a) from body fluids sample of said individual, preferably blood sample, evaluating the plasma or serum biomarkers of inflammation resolution, said plasma or serum biomarkers comprising at least a set of pro-inflammatory biomarkers and a set of pro-resolving biomarkers, b) determine the resolution of inflammation index of said individual thanks to comparison of the balance between pro-inflammatory and pro-resolving biomarkers between two measurements, wherein each measurement is separated by a minimum of at least 3 weeks , preferably 1 month, more preferably 1,5 month, and preferably, and wherein a first dietary supplement containing SPMs and/or their precursors issued from omega 3 (EPA, DHA) rich oil, is taken by said individual during these two separated measurements,

c) define a composition of a second personalized dietary supplement necessary to increase said resolution of inflammation index of said individual, preferably based on the value of the production of pro-resolving mediators versus pro-inflammatory mediators obtained in step b), and

d) ingestion by said individual of said second personalized dietary supplement.

In the methods according of the present invention, said resolution index is calculated as followed : [selected SPMs and/or precursors of pro-resolving mediators] / [selected pro- inflammatory mediators].

As instance it has been evaluated on two individuals after food high level SPM food supplementation and given the following results (see table 2): Table 2: resolution of inflammation index evaluated on two individuals after food high level SPM food supplementation

In a preferred embodiment of the methods of the present invention, the evaluation of a set of pro-resolving mediators biomarkers of inflammation resolution in the plasma or serum comprises the determination of the plasma or serum levels of at least 5, preferably 7, 9, or all of the SPMs and their precursors selected from the group consisting of:

LXA4 (lipoxin A4), LXB4 (lipoxin B4), RVE1, RVE2, RVE3 (resolvins El, E2, E3), RVD1, RVD2, RVD3, RVD4, RVD5, RYD6 (Resolvins Dl , D2, D3, D4, D5, D6), PD1 (protectin Dl), MaRl and/or MaR2 (maresin 1 and/or 2).

In a further preferred embodiment of the method of the present invention, the evaluation of a set of pro-inflammatory and a set of pro-resolving mediators biomarkers of inflammation resolution in the plasma or serum comprises the determination of the plasma or serum level of at least the following 9 molecules:

PGE2, LTB4, TXB2, 5HETE, 12HETE, 15HETE, l7HDoHE, MHDoHE and 18HEPE.

In a further preferred embodiment of the method of the present invention the evaluation of a set of pro-inflammatory and a set of pro-resolving mediators biomarkers of inflammation resolution in the plasma or serum comprises the determination of the plasma or serum level of all the following pro-resolving mediators l7HDoHE, 14HDoHE, 18HEPE, 12HETE, 15HETE, LXA4, LXB4, RVE1, RVE2, RVE3, RVD1, RVD2, RVD3, RVD4, RVD5, RVD6, PD 1, MaRl and MaR2. In a preferred embodiment, the sample taken from the patient may be selected from body fluid sample, such as blood, urine or sweat sample, or tissue sample of a patient.

In a more preferred embodiment, said patient sample is a blood sample, particularly selected from whole blood, serum or plasma sample. Plasma is the most preferred.

In another embodiment, in step a) of the method according to the present invention, the patient's blood sample may be treated prior to the measurements being taken. For example, step a) is carried out on blood sample, particularly in plasma sample obtained after activating cells with a stimulus as instance immune system ligands as toll-like receptors (TLR) ligands or bacteria/fungus cell wall or inductors of signal transduction such as phorbol 12-myristate 13- acetate (PM A) or calcium ionophore.

Step a) is metabolo-lipidomic technic for the quantification of biomarkers of resolution to establish in step b) a personal resolution index.

In a preferred manner, the method according to the present invention is characterized in that it further comprises the following step:

e) repeat steps a) to d) until said resolution of inflammation index of said individual is minimally improved by 50%. s

This new solution consists in a selection of food ingredients that will improve the level of circulating SPMs in the blood when they will be eaten and measuring circulating SPMs to enter into a positive loop of amelioration and personalisation of the food supplementation. We have called this concept an Objectified Personalized Nutrition solution (OPeN).

This method will give the proof to its customers (i.e. said individual) that the intake of ingredients containing SPMs induces higher amounts of SPMs in the blood. Based on the idea that uncontrolled and low-grade inflammation are at least in part due to a defect of resolution, higher amount of SPMs will restore the natural capacity of the body to prevent inflammation.

To our knowledge no company worldwide integrates both a food supplementation and a blood evaluation of inflammation or its resolution to improve the supplementation.

None company worldwide using dietary supplements measure that the body has effectively incorporated their products. The association of our metabolo-lipidomic test with the individual supplementation will permit this. Our strategy of OPeN is unique at the moment and will contribute to the knowledge within inflammation especially in the arrest of that response (resolution).

In the method of the present invention, the step a) comprises the evaluation in the patient sample of the concentrations of the selected pro-inflammatory mediators, their precursors, SPMs and their precursors for determining the resolution of inflammation index of said individual thanks to comparison of the balance between pro-inflammatory and pro-resolving biomarkers.

To measure the resolution inflammation index, it has been implemented, in a preferred manner, a technique that allows the measurement of those metabolites and which requires a high expertise: the liquid chromatography coupled to mass spectrometry in tandem (LC- MS/MS). The identification of the network of lipid biomarkers thanks to this approach is termed metabolo-lipidomics. Other techniques known from the man state in the art, allowing the same results, can be used according to the present invention.

In a preferred manner, the method according to the present invention is characterized in that step a) said concentrations evaluation of each selected biomarker is performed by liquid chromatography coupled to mass spectrometry in tandem (LC-MS/MS) or gas chromatography.

In an also preferred embodiment, the method according to the present invention is characterized in that step a) said concentrations evaluation is performed by specific immunoassays, particularly enzyme immunoassays.

Antibody reagents specifically directed against individual lipid mediator are generally known to those skilled in the art. These antibodies can be labelled with enzymes, radioisotopes, and fluorescent, luminescent and chromogenic substances, including coloured particles such as colloidal gold or latex beads. Suitable immunoassays include enzyme-linked immunosorbent assays (ELISA). Various types of labels and methods of conjugating the labels to the antibodies are well known to those skilled in the art.

In a second aspect, the present invention is directed to a dietary supplement characterized in that it comprises from intermediate to high level of SPMs issued from omega 3 (EPA, DHA) rich oil or that the dietary supplement is able to induce the production of high levels of SPMs by tissues during inflammation.

In a preferred embodiment, by high level of SPMs or their precursors issued from omega 3 (EPA, DHA) rich oil, it is intended to designate dietary complement contained at least one, preferably 2, more preferably the 3 following SPMs precursors (see table 1).

14-HDOHE and 17-HDOHE are the most preferred.

SPMs according to the present invention are lipoxins A4 and B4 (LXA4 and LXB4), resolvines of type E (RVE1; RVE2 and RVE3), resolvines of type D (RVD1 ; RVD2; RVD3; RVD4; RVD5; RVD6), protectin Dl (PD1) and maresins (MaRl and MaR2), plus their precursors issued from EPA (18-HEPE) or DHA (17-HDOHE, 17-HDOHE) or arachidonic acid (15-HETE, 12-HETE).

As instance if a defect of resolvins of type E is analysed then food supplementation would preferably contain intermediate or high levels of 18-HEPE. If, type D resolvins, and/or PD1 and or Marl and/or Mar2 are analysed to be defective then food supplementation would preferably contain intermediate or high level of 14-HDOHE and/or 17-HD0HE.

By the term high level of SPMs in a dietary supplement, it is intended to designate as instance levels of 18-HEPE over 38 563 pg/mg of crude oil; of 14-HDQHE over 32 568 pg/mg of crude oil; of 17-HDOHE over 48 524 pg/mg of crude oil.

Embodiments of the dietary supplement include various combinations of the SPMs instances described above. The described instances should thus not be construed as being mutually exclusive, and the inclusion of any one of the above-described instances will not preclude the inclusion of any other of the above-described instances.

In particular, said dietary supplement may comprise food raw ingredients containing high level of SPMs that can be easily introduce into capsules.

Preferably, said dietary supplement comprises fish oil containing SPMs. These oils can be incorporated into capsules to supplement a diet and provide the lacking mediators during inflammation.

In a preferred manner, said dietary supplement comprises micro-algae or krill, containing SPMs. Indeed, there are new emerging and natural sources of omega-3 with a reduced impact on the environment than fish oils (micro-algae, krill...). Microalgae may contain naturally SPMs or may be genetically engineered to improve this capacity.

In a third aspect, the present invention is directed to a kit for the resolution of inflammation in an individual, said kit comprising:

a) instructions to apply the method to increase the resolution of inflammation of an individual according to the present invention, in order to obtain the resolution of inflammation index of said individual

b) the dietary supplement according to the present invention. In a preferred embodiment, the dietary supplement of the kit according to the present invention is constitued of gelules or containers, each of them containing only one type of SPMs or of their precursors, or , in a further preferred embodiment, only 2 or 3 types of SPMs or their precursors at the maximum.

In a preferred embodiment, the dietary supplement of the kit according to the present invention comprised at least 2 compositions wherein the SPMs, or the combination of SPMs, contained in each composition are qualitatively and/or quantitatively different.

Indeed, the design of the dietary supplement according to the invention will be depending on the missing critical SPMs target identified by the biomarkers analysis.

The present invention is also directed to a kit useful for increasing the resolution of inflammation in an individual, said kit comprising the analysis reagents needed to obtain or measure the resolution of inflammation index of said individual and an algorithm which can be residing in a computer for correlating the index obtained with known reproductive inflammation events.

The correlating is performed by an algorithm that is trained to identify patterns in individual with the levels of said plasma or serum biomarkers of inflammation resolution in patient exhibiting uncontrolled inflammation and/or exhibiting inflammation which can be correctly stopped with secretion of SPMs.

In a preferred embodiment, the algorithm further correlates the measured resolution of inflammation index to the composition in SPMs of a personalized dietary supplement necessary to increase said resolution of inflammation index of said individual.

Finally, in a fourth aspect, the present invention is directed to the use of the dietary supplement according to the present invention, the method according to the present invention, or the kit of the present invention for the prevention of uncontrolled inflammation and/or for relief of the minor daily health trouble linked with inflammation.

Multiple benefits for the society and the environment can be identified for our products. We first believe that this OPeN will contribute to live older and in better health: this will thus decrease the cost on social system. A second benefit will be the reduction of the use of OTC drugs: this will decrease the pollution linked to anti-inflammatory drugs in the rivers that have been identified for many years.

Another important type of individuals targeted by the present invention is people suffering from chronic illnesses. Indeed, they used to take dietary supplements during long periods, generally advised by a therapist, in support of allopathic treatments.

Finally, an additional important trend to take in account is the preference for natural products instead of synthetic alternatives. Key points of the present invention will be the following:

- Restoration of the natural capacity of the body to end inflammation

- Personalization of the food supplementation and objectification of its effects.

The present invention consists in a blood-based test (as instance by metabolo-lipidomics analysis) that will deliver an individual index of resolution. In function of the resolution index, we will advise an adapted dietary supplement. Each advised supplementation will consist in capsules created by Ambiotis with an adapted dosage of SPMs issued from omega 3 (EPA, DHA) - rich oil. The same test should then be performed regularly to measure the benefit of the food supplementation. In fine, this results in an Objectified Personalized Nutrition (OPeN).

The present invention is based on:

1) The capacity to distinguish a good inflammation (that will solve correctly) from a bad one (which stopping is delayed or abolished) thanks to the measurement of the balance between pro-inflammatory and pro-resolving biomarkers (resolution index). Thanks to this balance we can determine whether the inflammation will stop (pro-resolving mediators high/proinflammatory mediators low) or if it is likely to last longer or increase (pro-resolving mediators low/proinflammatory mediators high);

2) the capacity to evaluate in blood samples this resolution index: we thus offer to evaluate the status of this balance in individual’s blood in order to identify a potential imbalance and thus define the supplementation strategy to restore the healthy state. The present invention is also directed to a method for determining the effect and/or the efficacy of nutrient dietary supplements in inflammation in an individual, said method comprising:

a) determining the missing critical SPMs target identified by the method according to the present invention suspected to allow to increase the resolution of inflammation of said individual,

b) adding a nutrient dietary supplement containing said missing critical SPMs to the patient diet,

c) performing the resolution of inflammation index of said individual after ingestion of said nutrient dietary supplement

d) assessing and/or comparing the resolution of inflammation index obtained in step c) and determining the effect and/or the efficacy of said supplement on the increase of the index.

The following examples and the figures and the legends hereinafter have been chosen to provide those skilled in the art with a complete description in order to be able to implement and use the present invention These examples are not intended to limit the scope of what the inventor considers to be its invention, nor are they intended to show that only the experiments hereinafter were carried out.

Other characteristics and advantages of the invention will emerge in the remainder of the description with the Examples and Figures, for which the legends are given hereinbelow.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1: Figure 1 shows a schematic representation of an acute inflammation, which is an auto-resolutive inflammatory process that starts, amplifies and finally end. Those three steps are deciphered by the plain curve. The resolution process (which allows the ending of the inflammatory response) is a complex process that involves the sequential and chronological engagement of cellular (eg granulocytes or macrophages) and chemical (eg cytokines or specialized pro-solving mediators) effectors.

In terms of bioactive lipid mediators, pro-inflammatory mediators are synthesized mainly during the beginning and the amplification steps of inflammation (dotted curve). Their decrease is due to the destruction (catabasis) of those mediators by the tissue that is the site of the inflammatory process. Prototypic inflammatory mediators are PGE2 and LTB4, issued from the arachidonic acid and some intermediates such as 5-HETE or PGG2.

To counterbalance the pro-inflammatory mediators, Omega3 fatty acids (as instance DHA or EPA) are mobilized to actively end inflammation and starts the resolution program of inflammation. This is possible thanks to the production of Specialized Proresolving Mediators or SPMs (dashed curve) such as Protecin, Maresins, Type D resolvins, type E resolvins, or their intermediates such as 14-HODHE, 17-HDOHE or 18-HEPE. Figure 2: Figure 2 shows the different steps implemented in the method to increase the resolution of inflammation of an individual according to the present invention.

A first blood collection will be done to evaluate the resolution index of the individual thanks to a metabololipidomics analysis of pro-inflammatory mediators and Specialized Proresolving mediators. A personalized strategy will be proposed to the consumer that corresponds to dietary supplementation with SPMs inside. The individual will then enter in an amelioration loop consisting in measurements of resolution index along the treatment, in order to adapt and/or personalize the quantity and/or the family of SPMs that will be delivered to the individual by pills. EXAMPLES

Two subjects have been involved in an Objectified and Personalized Nutritional intervention (OPeN). They have received 2 000 mg/day during 30 days of rich oil containing high level of SPM (18-HEPE : 45 636 pg/mL; 14-HDOHE : 35 689 pg/mL; 17-HDOHE : 56 897 pg/mL). Before and after starting the food supplementation SPMs and proinflammatory mediators have been measured in the plasma in order to calculate their respective resolution indices. More precisely, SPMs and proinflammatory mediators were analysed by mixing the plasma with methanol and freezing at -80°C. After thawing, samples were taken to solid phase extraction in the presence of deuterated internal standards and lipid mediators (LM) eluted in HCOOMe. After solvent evaporation, samples were dissolved in MeOH and injected into an Agilent 1290 Infinity high performance liquid chromatography (HPLC) system equipped with a Kinetex Biphenyl column (2.1 mm, 50 mm, 1.8 pm) (Phenomenex). LM were eluted with a binary gradient of water/formic acid 0.1 % and acetonitrile/formic acid 0.1 % and taken to MS/MS analysis on a triple quadrupole Agilent 6490 instrument. LM were identified based on matching of retention time to authentic standards. Calibration curves were obtained using authentic LM mixtures and quantification was carried out based on peak areas from multiple reaction monitoring (MRM) transitions.

Following Tables 2 and 3 present the contain of plasma of the said subjects before and after food supplementation :

The resolution index was calculated as followed : [selected SPMs and precursors of pro-resolving mediators] / [selected pro-inflammatory mediators].

Claims

1) Method to increase the resolution of inflammation of an individual, comprising the following steps of:

a) from body fluids sample of said individual, preferably blood sample, evaluating the plasma or serum biomarkers of inflammation resolution, said plasma or serum biomarkers comprising at least a set of pro-inflammatory biomarkers and a set of pro-resolving biomarkers,

b) determine the resolution of inflammation index of said individual thanks to comparison of the balance between pro-inflammatory and pro-resolving biomarkers, c) define the composition in specialized pro-resolving mediators (SPMs), of a personalized dietary supplement necessary to increase said resolution of inflammation index of said individual, and

d) ingestion by said individual of said personalized dietary supplement.

2) Method according to claim 1, characterized in that it further comprises the following step:

e) repeat steps a) to d) until said resolution of inflammation index of said individual is satisfying.

3) Method for determining or to predict whether an inflammation in an individual can be stopped or if said inflammation is likely to last longer or increase, said method comprises the following steps:

b) determining the resolution of inflammation index of said individual thanks to comparison of the balance between pro-inflammatory and pro-resolving biomarkers, and c) determine whether the inflammation will stop or if it is likely to last longer or increase, wherein:

- high level of pro-resolving mediators versus low level of pro-inflammatory mediators is significant of inflammation which can be stopped, and

- low level of pro-resolving mediators versus high level of pro-inflammatory mediators is significant of inflammation which is likely to last longer or increase.

4) Method for determining or to predict whether an inflammation in an individual is susceptible to be stopped by the ingestion of dietary supplement containing SPMs, said method comprises the following steps:

b) determining the resolution of inflammation index of said individual thanks to comparison of the balance between pro-inflammatory and pro-resolving biomarkers, and

c) determine whether the inflammation is susceptible to be stopped by the ingestion of dietary supplement containing SPMs, wherein:

- high level of pro-resolving mediators versus low level of pro-inflammatory mediators is significant of inflammation which is susceptible to be stopped by the ingestion of dietary supplement containing SPMs, and

- low level of pro-resolving mediators versus high level of pro-inflammatory mediators is significant of inflammation which is not susceptible to be stopped by the ingestion of dietary supplement containing SPMs.

5) Method for determining or to predict whether resolution in an individual, suffering of an uncontrolled inflammation, can be improved or if said uncontrolled inflammation is likely to last longer or increase, preferably whether the resolution of inflammation can be improved by the ingestion of dietary supplement containing SPMs or selected compounds that have been tested to improve SPM production during uncontrolled inflammation, said method comprises the following steps : a) from body fluids sample of said individual, preferably blood sample, evaluating the plasma or serum biomarkers of inflammation resolution, said plasma or serum biomarkers comprising at least a set of pro-inflammatory biomarkers and a set of pro-resolving biomarkers,

b) determining the resolution of inflammation index of said individual thanks to comparison of the balance between pro -inflammatory and pro-resolving biomarkers, and

- no or less production of pro-resolving mediators versus same or higher level of pro- inflammatory mediators is significant of inflammation which is likely to last longer or increase;

- each measurement is separated by a minimum of at least 3 weeks , preferably 1 month, more preferably 1,5 month, and preferably,

6) The method according to one of claims 1 to 5, wherein in step a), the plasma or serum level or concentration of at least one biomarker of inflammation resolution is evaluated in each of these following 3 biomarkers groups, these 3 groups being consisting of

- pro inflammatory PGE2 (prostaglandin E2), LTB4 (leukotriene B4), TXB2 (thromboxane B2) biomarkers,

- their precursors 5HETE, 12HETE, 15HETE (5-, 12-, 15-Hydroxyeicosatetraenoic acid),

- precursors of pro-resolving mediators 17HDoHE (l7-Hydroxy-4,7,l0,l3,l5,19- docosahexaenoic acid), !4HDoHE (!4-Hydroxy-4Z,7Z,l0Z,l2E,l6Z,19Z- docosahexaenoic acid), 18HEPE (l8-hydroxy-5Z,8Z,HZ,l4Z,l6E-eicosapentaenoic acid).

7) The method according to one of claims 1 to 6, wherein in step a), the plasma or serum level or concentration of the following 9 biomarkers is determined:

PGE2, LTB4, TXB2, 5HETE, 12HETE, 15HETE, l7HDoHE, MHDoHE and 18HEPE.

8) Method according to one of claims 1 to 7, characterized in that step a) is performed by a method selected from the group consisting of liquid chromatography coupled to mass spectrometry in tandem (LC-MS/MS), gas chromatography and immunoassays.

9) Method for determining the effect and/or the efficacy of nutrient dietary supplements in inflammation in an individual, said method comprising:

a) determining the missing critical SPMs target identified or defining the composition in SPMs of a personalized dietary supplement necessary to increase said resolution of inflammation index of said individual by the method according to one of claims 3 to 7,

b) adding a nutrient dietary supplement containing said missing critical SMPs or the personalized dietary supplement to the patient diet,

c) performing the resolution of inflammation index of said individual after ingestion of said nutrient dietary supplement, and

10) Dietary supplement characterized in that it comprises high level of SPMs issued from omega 3 rich oil, particularly from eicosapentaenoi^'c acid (EPA) and/or docosahexano^'ic acid (DELA) rich oil. 11) Kit for the resolution of inflammation in an individual, said kit comprising:

a) instructions to apply the method according to one of claims 1 to 9, in order to obtain the resolution of inflammation index of said individual b) the dietary supplement according to claim 9.

12) Kit useful for increasing the resolution of inflammation index in an individual, said kit comprising:

- the analysis reagents needed to obtain or measure the resolution of inflammation index of said individual, and

- an algorithm which can be residing in a computer for correlating the index obtained with known reproductive inflammation events. 13) Kit according to claim 12, wherein the algorithm further correlates the measure of the resolution of inflammation index to the composition in SPMs of a personalized dietary supplement necessary to increase said resolution of inflammation index of said individual.

14) Use of the method according to one of claims 1 to 9, the dietary supplement according to claim 10, or the kit according to one of claims 11 to 13, for the prevention of uncontrolled inflammation.

15) Use of the method according to one of claims 1 to 9, the dietary supplement according to claim 10, or the kit according to one of claims 11 to 13, for the prevention of uncontrolled inflammation for relief of the minor daily health trouble linked with inflammation.