Extract from Combretum micranthum G. Don

Abstract

The present invention relates to an extract of Combretum micranthum G. Don, for use in human or animal subjects in the treatment and / or prevention of pain. The present invention further relates to a pharmaceutical or non-pharmaceutical composition containing an extract of Combretum micranthum G. Don according to the invention, for use in human or animal subjects in the treatment and / or prevention of pain.

Description

Title of the invention: Extract of Combretum micranthum G. Don Technical field of the invention

[0001] The present invention relates to the technical field of plant extracts, for their use in human or animal subjects, in the treatment and / or prevention of pain. State of the art

[0002] Currently, the drug management of chronic pain conditions has limitations.

[0003] The misuse of pain-related medications is a very significant problem. This misuse concerns the most commonly used painkillers such as paracetamol, aspirin, anti-inflammatories or morphine and its derivatives.

[0004] Analgesics are considered active and effective against acute pain but have significant side effects if used for a long time: gastric and renal disorders, dependence, habituation, etc.

[0005] Thus these drugs are not without side effects and are therefore not suitable for chronic use or for the management of chronic pain.

[0006] For the latter, particularly neuropathic pain, the treatments are antidepressants and antiepileptics, which have fewer side effects but with observable efficacy in only 50% of patients.

[0007] There is therefore a need to develop effective, safe and well-tolerated treatments for patients.

[0008] In this context, many consumers are turning to natural approaches based on plants or plant extracts to improve their quality of life.

[0009] Plant extracts indeed offer potential to meet this need due to their effectiveness, low toxicity and availability.

[0010] Accordingly, there is continued interest in developing novel plant extracts suitable for the treatment and prevention of pain. Presentation of the invention

[0011] In order to overcome the aforementioned drawback of the state of the art, the present invention provides an extract of Combretum micranthum G. Don, for its use in human or animal subjects, in the treatment and / or prevention of pain.

[0012] Indeed, surprisingly, the present invention provides a plant ingredient, derived from Combretum micranthum G Don, for treating or preventing pain in human or animal subjects, with an interesting efficacy profile.

[0013] Furthermore, this plant ingredient according to the invention is advantageously standardized in terms of phytochemical composition and biological activity.

[0014] Such a plant ingredient also constitutes a natural resource composed of a mixture of active molecules (versus a single synthetic molecule), suitable for chronic intake.

[0015] Other non-limiting and advantageous characteristics of the product / process according to the invention, taken individually or in all technically possible combinations, are the following:

[0016] - said pain is chosen from joint, visceral, pelvic pain, post-operative, inflammatory, neuropathic, arthritic;

[0017] - the treatment and / or prevention of pain is obtained by means of a anti-inflammatory effect;

[0018] - the treatment and / or prevention of pain includes a reduction of its intensity and / or a reduction in its duration;

[0019] - the pain is chosen from mild pain, moderate pain, and severe pain;

[0020] - said extract comes from an extraction from the leaves;

[0021] - said extract contains kinkeloids, namely advantageously chosen from among the kinkeloids A, kinkeloids B, kinkeloids C and kinkeloids D.

[0022] Still generally, the extract of Combretum micranthum G. Don according to the invention preferably consists of an extract resulting from a process chosen from the following processes:

[0023] - a methanolic extraction process comprising the following successive steps:

[0024] — a step of grinding the plant material,

[0025] — a step of macerating the crushed plant material in an alcohol, for example methanol,

[0026] — a stage of filtration of the macerates,

[0027] — a step of collecting the filtrates and evaporating them to remove the alcohol and water,

[0028] — a freeze-drying step;

[0029] - a process for preparing a plant extract, from plant material, including:

[0030] — a step of contact between the plant material and a solvent,

[0031] — at least one step of liquid phase extraction of the active ingredients, carried out under heating and under reduced pressure, to obtain an extraction fraction containing said active ingredients and residual plant material, and

[0032] — a step of drying said residual plant material in the presence of said extraction fraction, in order to allow the fixation of the active ingredients on said material residual plant,

[0033] - a process for preparing a plant extract comprising:

[0034] — a step of soaking a ground plant material, in the presence of a solvent al alcoholic, for example ethanol,

[0035] — a percolation step by adding an alcoholic solvent, for example ethanol,

[0036] — a re-soaking step by adding an alcoholic solvent, for example ethanol,

[0037] — a step of filtration of the re-soaking solvent,

[0038] — a step of concentration of the filtrate and lyophilization,

[0039] - a process for preparing a plant extract comprising:

[0040] — a step of soaking the ground plant material, in the presence of a solvent al alcoholic, for example ethanol,

[0041] — a percolation step by adding an alcoholic solvent, for example ethanol,

[0042] — a re-soaking step by adding an alcoholic solvent, for example ethanol,

[0043] — a step of filtration of the re-soaking solvent,

[0044] — a step of concentrating the filtrate by evaporation,

[0045] — a step of stabilizing the dry extract with alcohol,

[0046] — a step of concentrating the plant extract, and

[0047] — a step of stabilization with glycerin of the extract.

[0048] The present invention also encompasses the aforementioned methods for obtaining the extract of Combretum micranthum G. Don according to the invention.

[0049] The present invention also relates to a composition containing an extract of Combretum micranthum G. Don according to the invention, for its use in human or animal subjects, in the treatment and / or prevention of pain.

[0050] Said composition advantageously further contains at least one of the following ingredients: an algae extract, a plant extract and probiotics, vitamins, minerals or any other nutritional ingredient.

[0051] This composition is advantageously suitable for use in the pharmaceutical, cosmetic, agri-food, nutrition, food supplement and animal nutrition industries.

[0052] The present invention also relates to a non-therapeutic use of an extract of Combretum micranthum G. Don according to the invention, for its use in human or animal subjects, in the treatment and / or prevention of pain.

[0053] Of course, the various features, variants and embodiments of the invention can be combined with each other in various combinations. to the extent that they are not incompatible or mutually exclusive. Detailed description of the invention

[0054] In addition, various other features of the invention emerge from the attached description.

[0055] The present invention therefore relates, in general, to extracts of Combretum micranthum G. Don, for their use in human or animal subjects, in the treatment and / or prevention of pain.

[0056] By "human subjects" is meant in particular patients suffering from chronic or acute pain. Human subjects may also include healthy individuals who wish to prevent pain related to their work, lifestyle, age, or individuals who have undergone medical intervention (e.g., surgery or dental work) and suffer from post-operative pain.

[0057] By "animal subjects" is meant in particular companion animals such as dogs and cats, which may suffer from chronic pain related to joint diseases or other conditions, as well as livestock such as cattle, pigs, sheep and equines.

[0058] The term "pain treatment" refers to use to reduce, relieve, or eliminate pain.

[0059] The term "pain prevention" refers to use to prevent or reduce pain before it occurs. Combretum micranthum G. Don

[0060] Combretum micranthum or Combretum micranthum G. Don is an undomesticated shrub species found in the jungles of West Africa.

[0061] It is also designated Bureava crotonoides, Combretum altum, Combretum flo-ribundum, Combretum parviflorum, Combretum raimbaultii or even kinkéliba (vernacular name).

[0062] It is a dense shrub or vine, commonly found in cultivated and fallow fields, mainly located in sub-Saharan Africa, with higher yields in Senegal, Mali and Burkina Faso.

[0063] In several countries of the tropical savannah of West Africa, the leaves of Combretum micranthum are used as a popular traditional herbal tea, particularly for its diuretic properties.

[0064] Still generally, Combretum micranthum G. Don is still defined in the Monograph of the French Pharmacopoeia, 1993 edition.

[0065] Extract of Combretum micranthum G. Don and its extraction processes

[0066] Preferably, the extract of Combretum micranthum G. Don is obtained from an extraction process from the leaves of Combretum micranthum G. Don.

[0067] The leaves of Combretum micranthum G. Don are advantageously fresh leaves or dried leaves.

[0068] Generally speaking, different extraction methods are suitable for obtaining an extract of Combretum micranthum G. Don according to the invention.

[0069] According to a first embodiment, the extraction process consists of a methanolic extraction process comprising the following successive steps:

[0070] - a step of grinding the plant material,

[0071] - a step of maceration of the crushed plant material in an alcohol, for example methanol,

[0072] - a stage of filtration of the macerates,

[0073] - a step of collecting the filtrates and evaporating them to remove the alcohol and water,

[0074] - a freeze-drying step.

[0075] Such a process is further developed in the following document: M. Gainche et al., Anti-inflammatory and cytotoxic potential of new phenanthrenoids from Luzula sylvatica, Molecules, 2020, 25 (10), 2372).

[0076] According to a second embodiment, the extraction method consists of a method for preparing a plant extract, from plant material, comprising:

[0077] - a contact step between the plant material and a solvent,

[0078] - at least one step of liquid phase extraction of the active ingredients, carried out under heating and under reduced pressure, to obtain an extraction fraction containing said active ingredients and residual plant material, and

[0079] - a step of drying said residual plant material in the presence of said extraction fraction, in order to allow the fixation of the active ingredients on said residual plant material.

[0080] Such a method is for example described in document EP-2 080 436.

[0081] According to a third embodiment, the extraction method consists of a method for preparing a plant extract comprising:

[0082] - a step of soaking a ground plant material, in the presence of a solvent al alcoholic, for example ethanol,

[0083] - a percolation step by adding an alcoholic solvent, for example ethanol,

[0084] - a re-soaking step by adding an alcoholic solvent, for example ethanol,

[0085] - a step of filtration of the re-soaking solvent,

[0086] - a step of concentration of the filtrate and lyophilization.

[0087] According to a fourth embodiment, the extraction method consists of a method for preparing a plant extract comprising:

[0088] - a step of soaking the ground plant material, in the presence of a solvent al alcoholic, for example ethanol,

[0089] - a percolation step by adding an alcoholic solvent, for example ethanol,

[0090] - a re-soaking step by adding an alcoholic solvent, for example ethanol,

[0091] - a step of filtration of the re-soaking solvent,

[0092] - a step of concentrating the filtrate by evaporation,

[0093] - a step of stabilizing the dry extract with alcohol,

[0094] - a step of concentrating the plant extract, and

[0095] - a step of stabilization with glycerin of the extract.

[0096] Still generally, the extract of Combretum micranthum G. Don according to the invention contains kinkeloids, compounds of the piperidine-flavan alkaloid family.

[0097] Kinkeloids are for example described in the document Jing Zhen et al., Total Synthesis of Novel Skeleton Flavan-Alkaloids, Molecules 2020, 25(19), 4491.

[0098] By “kinkeloids” we mean in particular kinkeloids A, kinkeloids B, kinkeloids C and kinkeloids D.

[0099] By kinkeloids A is meant the chemical compounds having the following chemical formula 1:

[0100] [Chem.l] ÔIH

[0101] in which

[0102] is H,

[0103] R2estH,

[0104] R3estH,

[0105] and in which:

[0106] R4cst H and R5 is piperidine, or

[0107] R4 is piperidine and R5 is H.

[0108] By kinkeloids B is meant the chemical compounds having the following chemical formula 2:

[0109] [Chem. 2] OH

[0110] [YES]

[0112]

[0113]

[0114]

[0115]

[0116]

[0117]

[0118] in which Ri is OH, R2 is H, R3 is H, and in which R4 is H and R5 is piperidine, or R4 is piperidine and R5 is H. Kinkeloids C are chemical compounds with the following chemical formula 3: [Chem. 3] OH

[0119]

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127] in which Ri is OH R2 is OH R3 is H, in which R4 is H and R5 is piperidine, or R4 is piperidine and R5 is H. Kinkeloids D are chemical compounds with the following chemical formula 4: [Chem. 4] R,

[0128] in which

[0129] R! is OH

[0130] R2 is OH

[0131] R3 is OH,

[0132] in which

[0133] R4 is H and R5 is piperidine, or

[0134] R4 is piperidine and R5 is H.

[0135] Preferably, the extract according to the invention contains kinkeloids B which represent from 70 to 90% of the total kinkeloids.

[0136] Without being limiting, a preferred dosage could be 20 mg to 5 g of extract according to the invention, per day, in one or more doses. Treatment and / or prevention of pain

[0137] By “pain” we mean in particular joint, visceral, pelvic, post-operative, inflammatory, neuropathic and arthritic pain.

[0138] The term "joint pain" refers to pain or discomfort felt in one or more joints of the body. More particularly, the invention aims to provide a solution for the treatment and / or prevention of joint pain, which can be caused by a variety of factors such as injuries, inflammation, diseases, or normal age-related wear and tear. Joint pain can affect any part of the body, including the hands, wrists, elbows, shoulders, hips, knees, ankles, and feet. Symptoms may include constant or intermittent pain, stiffness, swelling, redness, or warmth around the affected joint, as well as loss of mobility and flexibility.

[0139] The term "visceral pain" or "pelvic pain" refers to pain or discomfort felt in the internal organs of the body, such as the stomach, intestines, gallbladder, liver, pancreas, kidneys, or lower abdomen. Visceral or pelvic pain can be caused by conditions such as ulcers, infections, inflammations, stones, or functional disorders.

[0140] The term "postoperative pain" refers to the pain and discomfort that occurs after a surgical procedure. Postoperative pain is a common phenomenon that can vary in intensity and / or duration depending on the type of procedure, individual sensitivity, and other factors. Postoperative pain may be localized to the incision site or may be felt in other parts of the body.

[0141] Inflammatory pain, associated with inflammation phenomena that persist abnormally, particularly includes joint pain. In this case, chronic activation of pain fibers leads to their sensitization, which then generalizes to the entire pain system. Also, even when treating the cause peripherally, the system can remain hyper-reactive.

[0142] Neuropathic pain is generally linked to damage to the central or peripheral nervous system (nerve damage, injury, etc.), the spinal cord, linked to amputations or a stroke. These lesions directly affect the pain detection system: they make the alarm system faulty and uncontrollable by conventional analgesics.

[0143] The term "arthritic pain" refers to the pain and inflammation that occurs in joints affected by arthritis. Arthritis is a disease characterized by inflammation of the joints, which can lead to pain, stiffness, reduced joint mobility, and other symptoms. Arthritis pain can vary in intensity, duration, and frequency depending on the severity of the disease and individual sensitivity.

[0144] Preferably, the extract of Combretum micranthum G. Don provides treatment and / or prevention of pain through an anti-inflammatory effect.

[0145] Generally, by "treatment and / or prevention of pain", the present invention encompasses a reduction in its intensity and / or duration.

[0146] By "intensity" of pain, we include pain measured by tools to characterize and evaluate it.

[0147] Generally speaking, questionnaires and pain scales make it possible to describe the manifestations of pain, and to measure the intensity as well as the impact on quality of life.

[0148] For adults, the most commonly used scale is the numeric scale or visual analogue scale, graduated from 0 for no pain, to 10 for the maximum pain imaginable.

[0149] For children, doctors often use a scale with faces.

[0150] Concerning neuropathic pain, two scales allow respectively to diagnose this type of pain (DN4) and to evaluate its intensity (NPSI).

[0151] Generally, the present invention encompasses a decrease in the intensity and / or duration for pain measured in a subject receiving the extract of Combretum micranthum G. Don according to the invention compared to the intensity and / or duration for pain measured in a subject not receiving the extract of Combretum micranthum G. Don according to the invention.

[0152] Pain includes in particular:

[0153] - mild pain,

[0154] - moderate pain, and

[0155] - severe pain.

[0156] Mild pain is pain of little intensity that can be easily supported by the subject and do not affect or only slightly affect his daily activity.

[0157] Moderate pain is more intense and can affect the subject's daily life, for example by limiting their movements or their ability to work.

[0158] Severe pain is the most intense pain, which can be disabling and prevent the subject from leading a normal life. Composition

[0159] The present invention also relates to compositions containing an extract of Combretum micranthum G. Don according to the invention, for its use in human or animal subjects, in the treatment and / or prevention of pain.

[0160] Such a composition advantageously also contains at least one of the following ingredients:

[0161] - an algae extract,

[0162] - a plant extract and

[0163] - probiotics, including postbiotics.

[0164] The algae extract can be chosen from different species of algae, such as porphyra, a red algae also known as nori, spirulina, chlorella or a brown algae.

[0165] The plant extract can come from different plants, such as boswellia, turmeric, blackcurrant leaves, willow, meadowsweet, horsetail, devil's claw, figwort, Para cress (Acmella oleracea).

[0166] Generally speaking, by "probiotic strain" we include in particular living microorganisms which, when ingested in sufficient quantity, exert positive effects on health, beyond traditional nutritional effects.

[0167] The probiotic strain suitable for the invention is physiologically acceptable. In other words, this probiotic strain can be administered without risk to animals or humans.

[0168] Probiotic strains include in particular the genera lactobacilli (bacteria of the genus Lactobacillus), bifidobacteria (bacteria of the genus Bifidobacterium), streptococci (bacteria of the genus Streptococcus) or lactococci.

[0169] Again in general, specific examples of probiotic microorganisms are Bifidobacterium adolescentis, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium pseudocatenulatum, Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus casei (Shirota), Lactobacillus rhamnosus (GG strain), Lactobacillus brevis, Lactobacillus crispatus, Lactobacillus delbrueckii (subsp bulgaricus, lactis), Lactobacillus fermentum, Lactobacillus helveticus, Lactobacillus gallinarum, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus salivarius, Lactobacillus alimentarius, Lactobacillus curvatus, Lactobacillus casei subsp. casei, Lactobacillus sake Lactococcus lactis, Enterococcus (faecalis, faecium), Lactococcus lactis (subspp lactis or cremoris), Leuconstoc mesenteroid.es subsp dex-tranicum, Pediococcus acidilactici, Sporolactobacillus inulinus, Streptococcus salvarius subsp. Thermophilus, Streptococcus thermophilus, Staphylococccus camosus , Staphylococcus xylosus, Saccharomyces (cerevisiae or boulardii), Bacillus (cereus var toyo or subtilis), Bacillus coagulons, Bacillus licheniformis, Escherichia coli strain nissle, Propionibacterium freudenreichii and their mixtures.

[0170] More particularly, these are probiotic microorganisms from the group of lactic acid bacteria, such as Lactobacillus and / or Bifidobacterium. Illustrative of these lactic acid bacteria, mention may be made more particularly of Lactobacillus johnsonii, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus casei or Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium animalis, Bifidobacterium lactis, Bifidobacterium infantis, Bifidobacterium adolescentis or Bifidobacterium pseudoca-tenulatum and mixtures thereof.

[0171] Generally, said at least one probiotic strain is advantageously chosen from live probiotic strains or inactivated probiotic strains.

[0172] By “living” form, we mean a form with the capacity to multiply provided that it is placed in an environment conducive to the recovery of this capacity.

[0173] Thus, within the meaning of the present invention, the term living covers the so-called dormant state in which the microorganisms can be placed following a physicochemical treatment such as, for example, freeze-drying.

[0174] Generally speaking, the probiotic strain is advantageously used in a lyophilized form.

[0175] This type of formulation has the advantages of being easily accessible, easy to implement and not raising any difficulties and / or constraints in terms of storage. Furthermore, it is compatible with the conditioning of microorganisms in a dormant state.

[0176] This freeze-drying can be carried out according to conventional methods or according to a method described for example in document FR-3 103 827.

[0177] Furthermore, the term "inactivated" refers to strains that have undergone treatment aimed at killing them. Such treatments may consist, by way of non-limiting example, of autoclave treatment, ultrasound, high-pressure homogenization or even osmotic shock.

[0178] The inactivated strains are advantageously intact.

[0179] The probiotic strain is advantageously inactivated by heat, also called “Tyndallized probiotic strain”.

[0180] The concentration of probiotic strain in the composition is, for example, 107 to 1012 CFU per daily dose.

[0181] The concentration of probiotic strain in the composition may generally be an effective amount to obtain the desired effect. This amount may be determined by the person skilled in the art using his general knowledge and by standard tests. This amount may, for example, for topical application, in daily dose expressed in CFU, from approximately 106 to approximately 1012, preferably at least 106 to approximately 1010, preferably from approximately 108 to approximately 109. It may alternatively be from approximately 105 to approximately 1012 organisms / g of product.

[0182] The probiotic strain may further be included at a level of 0.01% to 50% in the composition, for example between 0.01% and 40%, or between 0.01% and 30%, or between 0.01% and 10%, or between 0.01% and 5%, or between 0.01% and 1%, for example about 0.3% (weight / weight).

[0183] The composition may also contain postbiotics, corresponding to preparations of microorganisms, advantageously chosen from the aforementioned probiotic strains, inanimate and / or their components conferring a benefit for the health of the host.

[0184] The composition may further contain other types of ingredients, for example vitamins, minerals or any other nutritional ingredient.

[0185] More particularly, the composition may contain nutraceutical ingredients such as vitamin C, glucosamine, chondroitin, palmitoylethanolamide or fatty acids.

[0186] The composition according to the invention advantageously contains a combination of ingredients suitable for use in the pharmaceutical, cosmetic, agri-food, nutrition (including foodstuffs intended for special medical purposes - DADFMS), food supplement and animal nutrition industries.

[0187] The composition according to the invention thus advantageously comprises:

[0188] - an extract of Combretum micranthum G. Don according to the invention, for its use in human or animal subjects, suitable for the treatment and / or prevention of pain, and

[0189] - at least one carrier or excipient, acceptable depending on the use considered.

[0190] The Combretum micranthum G. Don extract of the present invention may be prepared in the form of a pharmaceutical composition, which may be a formulation such as tablets, capsules, powders, granules, solutions, lozenges, jellies, cream preparations, syrups, suspensions, tinctures, aerosols and the like.

[0191] The Combretum micranthum G. Don extract of the present invention may also be prepared in the form of a non-pharmaceutical composition.

[0192] The non-pharmaceutical composition includes cosmetic compositions, agri-food compositions, nutritional compositions, food supplements, compositions for animal nutrition.

[0193] The non-pharmaceutical composition may be prepared by generally known preparation techniques, and acceptable additives may be added to the non-pharmaceutical composition.

[0194] For example, the non-pharmaceutical composition according to the invention advantageously consists of a food supplement.

[0195] By "food supplement" we advantageously mean food supplements which are subject to all the general provisions of food law but also to the specific rules defined by Directive 2002 / 46 / EC of the European Parliament and of the Council of 10 June 2002 on the approximation of the laws of the Member States concerning food supplements, transposed into French law by Decree No. 2006-352.

[0196] By "food supplements" is advantageously included foodstuffs whose purpose is to supplement the normal diet and which constitute a concentrated source of nutrients or other substances having a nutritional or physiological effect alone or in combination, marketed in the form of doses, namely presentation forms such as ampoules of liquid, bottles with a dropper and other similar forms of liquid preparations.

[0197] Generally, the composition according to the invention is suitable for oral, per os or topical administration. Non-therapeutic use

[0198] The present invention also encompasses a non-therapeutic use of an extract of Combretum micranthum G. Don according to the invention, for its use in human or animal subjects, in the treatment and / or prevention of pain.

[0199] Of course, various other modifications may be made to the invention within the scope of the appended claims. Material Examples

[0200] - Latin name: Combretum micranthum G.Don

[0201] - Plant part: leaves

[0202] - State of the plant: dry

[0203] Example 1 - extraction method according to the first embodiment

[0204] 1. Extraction protocol

[0205] - crush the plant material

[0206] - macerate the samples in methanol (10% w / v) 3 x 24h with stirring constant mechanics

[0207] - filter samples every 24 hours

[0208] - collect the filtrates after each extraction, evaporate the filtrates to remove the methanol

[0209] - freeze-dry.

[0210] Extraction yield in dry matter: 20 to 35%

[0211] - fractionate the methanol extract according to the protocol developed by Gainche et al. (M. Gainche et al., Anti-inflammatory and cytotoxic potential of new phenanthrenoids from Luzula sylvatica, Molecules, 2020, 25 (10), 2372).

[0212] - purify the fraction containing the kinkeloids.

[0213] This method made it possible to determine the quantity of kinkeloids in the methanolic extract: between 40 and 140 mg of kinkeloids per gram of methanolic extract.

[0214] 2. Writhing tests

[0215] The cramp test is a chemical method used to induce pain of peripheral origin by injecting irritants such as acetic acid into mice. The analgesic activity of the test compound is deduced from the reduction in the frequency of cramps.

[0216] Oral administration of the methanolic extract according to the invention, in a single dose of 300 mg / kg, 30 min before the injection of acetic acid, reduced the number of abdominal cramps. This result confirms the analgesic effect of this extract.

[0217] 3. AlgoGram (registered trademark)

[0218] The Algogram is a screening model proposed by ANS biotech to evaluate the analgesic activity of pharmaceutical molecules. It involves 11 animal models of pain to cover different areas of pain. It is useful for predicting on which type of pain a molecule / extract may be active.

[0219] The results show that the methanolic extract, administered per os at 300 mg / kg 30 minutes before pain assessment, confers an interesting effect on visceral, post-operative, inflammatory pain and in a model of neuropathic pain (hypersensitivity to cold induced by oxaliplatin).

[0220] 4. Anti-inflammatory activity in vitro

[0221] To investigate the mechanisms involved in the analgesic effects, the evaluation of the anti-inflammatory effects of the methanolic extract at different concentrations was carried out in an in vitro model of pro-inflammatory polarized macrophages. The plant extract was tested at 4 concentrations, and its effects were evaluated by ELISA tests to measure the production / decrease of pro-inflammatory cytokines on IL-1 [3, IL-6 and TNF-a, 3 major cytokines involved in inflammatory responses.

[0222] It is observed that the production of TNF-a was significantly reduced in the presence of the lowest concentration of plant extract, showing an anti-inflammatory effect of the plant extract.

[0223] 5. Conclusion for the methanolic extract

[0224] Overall, an analgesic activity of the methanolic extract according to the invention administered orally as a single dose (300 mg / Kg) could be demonstrated in two different screening models: the Writhing test (3 series) and the AlgoGram™. The latter shows a potential effect in different types of pain: visceral, post-operative, inflammatory and neuropathic pain. The data obtained in vitro on pro-inflammatory macrophages show that the methanolic extract according to the invention could exert its effects at least in part by an inhibition of the production of TNF-alpha.

[0225] Example 2 - extraction method according to the second embodiment

[0226] 1. Extraction protocol

[0227] - mixture of plant and water, heated for 30 minutes at 80°C;

[0228] - Concentration of the extract under vacuum;

[0229] - Spraying the extract on crushed Combretum leaves.

[0230] 2. AlgoGram

[0231] The present extract exhibits inhibition of visceral pain and pain related to kaolin-induced arthritis.

[0232] 3. Conclusion

[0233] This extract makes it possible to retain part of the analgesic activity demonstrated for the methanolic extract.

[0234] Example 3 - extraction method according to the third embodiment

[0235] 1. Protocol

[0236] - a step of soaking a ground plant material, in the presence of ethanol, before extensively heated,

[0237] - a percolation step by adding ethanol,

[0238] - a re-soaking step by adding ethanol,

[0239] - a step of filtration of the re-soaking solvent,

[0240] - a step of concentration of the filtrate and lyophilization, for example by evaporation.

[0241] 2. Writing tests

[0242] In the Writhing test, the extract according to the invention exhibits analgesic activity comparable to that of the methanolic extract with a reduction in the number of abdominal cramps.

[0243] 3. Algogram

[0244] The extract according to the invention administered orally as a single dose at a dose of 300 mg / Kg presents in the AlgoGram an analgesic profile similar to that of the methanolic extract with a lower amplitude of effect.

[0245] Inhibition is observed in the visceral pain model (TNBS) and in both inflammatory pain models (Kaolin and carrageenan, respectively).

[0246] 4. Conclusion

[0247] An analgesic activity of the extract according to the invention, administered orally as a single dose at a dose of 300 mg / kg, could be demonstrated in two different screening models.

[0248] The analgesic profile highlighted by the latter is similar to that of the methanolic extract although the amplitude of effect is lower.

[0249] Example 4 - extraction method according to the fourth embodiment

[0250] 1. Protocol

[0251] - a step of soaking the ground plant material, in the presence of ethanol,

[0252] - a percolation step by adding ethanol,

[0253] - a re-soaking step by adding ethanol,

[0254] - a step of filtration of the re-soaking solvent,

[0255] - a step of concentrating the filtrate by evaporation,

[0256] - a step of stabilizing the dry extract with alcohol,

[0257] - a step of concentrating the plant extract, and

[0258] - a step of stabilization with glycerin of the extract.

[0259] 2. AlgoGram

[0260] The extract according to the invention, administered orally as a single dose at a dose of 2000 mg / Kg, has an analgesic profile at AlgoGram similar to that of the methanolic extract with a less significant amplitude of effect.

[0261] An inhibition of visceral pain (TNBS model), and of inflammatory pain (Kaolin and carrageenan model), could be demonstrated.

Claims

Claims

1. Extract of Combretum micranthum G. Don, for use in human or animal subjects, in the treatment and / or prevention of pain.

2. Extract of Combretum micranthum G. Don, according to claim 1, characterized in that said pain is chosen from joint, visceral, pelvic, post-operative, inflammatory, neuropathic, arthritic pain.

3. Extract of Combretum micranthum G. Don, according to any one of claims 1 or 2, characterized in that the treatment and / or prevention of pain is obtained via an anti-inflammatory effect.

4. Extract of Combretum micranthum G. Don, according to any one of claims 1 to 3, characterized in that the treatment and / or prevention of pain includes a reduction in its intensity and / or its duration.

5. Extract of Combretum micranthum G. Don, according to any one of claims 1 to 4, characterized in that the pain is chosen from: - mild pain, - moderate pain, and - severe pain.

6. Extract of Combretum micranthum G. Don, according to any one of claims 1 to 5, characterized in that said extract comes from an extraction from the leaves.

7. Extract of Combretum micranthum G. Don, according to any one of claims 1 to 6, characterized in that said extract is obtained from a process chosen from the following processes: - a methanolic extraction process comprising the following successive steps: - a step of grinding the plant material, - a step of maceration of the ground plant material in an alcohol, for example methanol, - a step of filtration of the macerates, - a step of collection of the filtrates and evaporation to remove the alcohol and the water, - a freeze-drying step; - a process for preparing a plant extract, from plant material, comprising: — a step of contact between the plant material and a solvent, — at least one step of liquid phase extraction of the active ingredients, carried out under heating and under reduced pressure, to obtain an extraction fraction containing said active ingredients and a residual plant material, and — a step of drying said residual plant material in the presence of said extraction fraction, in order to allow the fixing of the active ingredients on said residual plant material, - a process for preparing a plant extract comprising: — a step of soaking a ground plant material, in the presence of an alcoholic solvent, for example ethanol, — a step of percolation by the addition of an alcoholic solvent, for example ethanol, — a step of re-soaking by adding an alcoholic solvent, for example ethanol, — a step of filtration of the re-soaking solvent, — a step of concentration of the filtrate and lyophilization,- a process for preparing a plant extract comprising: — a step of soaking the ground plant material, in the presence of an alcoholic solvent, for example ethanol, — a percolation step by adding an alcoholic solvent, for example ethanol, — a re-soaking step by adding an alcoholic solvent, for example ethanol, — a step of filtering the re-soaking solvent, — a step of concentrating the filtrate by evaporation, — a step of stabilizing the dry extract with alcohol, — a step of concentrating the plant extract, and — a step of stabilizing the extract with glycerin.,

8. Extract of Combretum micranthum G. Don, according to any one of claims 1 to 7, characterized in that said extract contains kinkeloids, advantageously chosen from kinkeloids A, kinkeloids B, kinkeloids C and kinkeloids D.

9. Pharmaceutical or non-pharmaceutical composition, containing an extract of Combretum micranthum G. Don according to any one of claims 1 to 8, for its use in human or animal subjects, in the treatment and / or prevention of pain.

10. A composition according to claim 9, further containing at least one of the following ingredients: an algae extract, a plant extract, probiotics, vitamins, minerals or any other nutritional ingredient.

11. Composition according to any one of claims 9 or 10, suitable for use in the pharmaceutical, cosmetic, agri-food, nutrition, food supplement and animal nutrition industries.

12. Composition according to claim 11, suitable for use in the cosmetic, agri-food, nutrition, food supplement, animal nutrition industry, in a non-therapeutic use of an extract of Combretum micranthum G. Don, for its use in human or animal subjects, in the treatment and / or prevention of pain.