Lipolytic and antiadipogenic coumarin derivatives

Abstract

The present invention relates to a composition comprising at least one compound represented by Formula (I) or a pharmaceutically acceptable salt thereof or a plant ex- tract comprising said at least one compound or said salt thereof for the use in the treatment of excess bodyweight in- cluding overweight and obesity and related metabolic and car- diovascular diseases, wherein R1 is -O-R4, R2 is -H or -OH and R3 is -O-R5, wherein R4 and R5 are independently an aliphatic C1 to C10 group, wherein the at least one compound represented by Formula (I) is administered to a subject in an amount of 200 µg / kg body weight to 8000 µg / kg body weight.

Description

[0001] LIPOLYTIC AND ANTIADIPOGENIC COUMARIN DERIVATIVES

[0002] TECHNICAL FIELD

[0003] The present invention relates to the field of coumarin and coumarin derivatives used in the treatment of diseases associated with lipolysis and adipogenesis .

[0004] BACKGROUND ART

[0005] Due to our modern li festyle that is characteri zed by overnutrition, physical inactivity, and stress , prevalence of obesity and obesity-induced diseases including diabetes , cardiovascular events , and cancer has been escalating world-wide within the last decades . These non-communicable diseases impose an enormous economic and health-political burden on society and necessitates the development of novel therapeutic concepts . In this context , the interest for dietary phytochemicals with preventive or therapeutic ef fects on obesity is enormous . From the customer ' s perspective , natural products are often preferred since they are received without medical prescription and are believed to exhibit lower risks of adverse side ef fects compared to synthetic drugs .

[0006] From a metabolic point of view, obesity originates from sustained energy imbalance between energy intake and energy expenditure . A positive balance over longer periods of time due to overnutrition, physical inactivity and genetic / epigenetic factors results in storage of excess energy into triglycerides ( TGs ) of white adipocytes . Development of mature lipid-loaden adipocytes from precursor cells is termed adipogenesis . Vice versa, under conditions of increasing energy demand due to excessive physical activity, fasting, and disease , TGs are enzymatically cleaved into free fatty acids ( FFAs ) and glycerol both serving as energy substrates .

[0007] The authors of Pan Y et al . ( J Food Biochem 46 ( 2022 ) : el 4200 ) investigated the influence of fermented and non- f ermented lemon peels on inflammatory processes in obese mice . It was found that the supernatant of fermented lemon peels may have positive ef fects on the regulation of blood lipids and liver function . The authors concluded that fermented lemon peel supernatants have anti-adipose and anti-inflammatory properties. IL-4, a cytokine that promotes lipolysis, is increased in mice treated with fermented lemon peel supernatants. The authors of Pay Y et al. found that the concentration of citropten in the supernatants is not increased by fermentation (it seems rather that fermentation reduces the concentration of citropten) , so that the observed effect with regard to IL-4 does not correlate with the citropten concentration.

[0008] Balwinder S et al. (Food Res Int 132 (2020) : 109114) discloses anti-obesity effects of citrus peel, which contains coumarins like limettin, bergamottin and 5-geranyloxy-7-methoxycoumarin .

[0009] WO 2012 / 094833 discloses extracts comprising 5,7- dimethoxycoumarin for treating glucose and lipid metabolism.

[0010] Peng S et al (Mediators Inflamm 2020 (2020) :8868107) discloses the beneficial effect of bergamot essential oil and juice against acne.

[0011] WO 03 / 059309 discloses cosmetic compositions comprising 5,7- dimethoxycoumarin . These compositions can be used as sunscreens .

[0012] It is an object of the present invention to provide methods and means to treat excess bodyweight including overweight and obesity and related metabolic and cardiovascular diseases.

[0013] SUMMARY OF THE INVENTION

[0014] The present invention relates to a composition comprising at least one compound represented by Formula (I) or a pharmaceutically acceptable salt thereof or a plant extract comprising said at least one compound or said salt thereof for the use in the treatment of excess bodyweight including overweight and obesity and related metabolic and cardiovascular diseases, wherein Ri is -O-R4, R2 is -H or - OH and Rs is -O-Rs, wherein R4 and Rs are independently an aliphatic Ci to C10 group, wherein the at least one compound represented by Formula ( I ) is administered to a subj ect in an amount of 200 pg / kg body weight to 8000 pg / kg body weight . The composition as defined above can also be used in the treatment of a disease or a disorder associated with decreased lipolysis and / or increased adipogenesis .

[0015] It turned surprisingly out that the compounds of the present invention are able to induce lipolysis . Lipolysis is the metabolic pathway through which lipid triglycerides are hydrolyzed into glycerol and free fatty acids . This pathway is useful to mobili ze stored energy within mammals from adipocytes , for instance , leading to a reduction of stored fat within these cells . The free fatty acids are typically released into the blood stream and made available for uptake to cells which require energy .

[0016] The increased lipolysis is beneficial in the treatment and / or prevention of excess bodyweight including overweight and obesity and related metabolic and cardiovascular diseases and of diseases and disorders associated with decreased lipolysis and / or increased adipogenesis , wherein adipogenesis involves the formation of adipocytes ( fat cells ) from stem cells .

[0017] It has been shown that the compound of the present invention and represented by Formula ( I ) when administered to a subj ect , in particular to a human subj ect , in the concentration range as defined herein, induces lipolysis . A too low concentration of the compound of the present invention has no or no signi ficant ef fect on lipolysis .

[0018] Another aspect of the present invention relates to a composition comprising at least one compound represented by Formula ( I )

[0019] ( I ) or a pharmaceutically acceptable salt thereof or a plant extract comprising said at least one compound or said salt thereof for the use in the treatment of excess bodyweight including overweight and obesity and related metabolic and cardiovascular diseases , wherein Ri is -O-R4 , R2 is -H or - OH and Rs is -O-R5, wherein R4 and Rs are independently an aliphatic Ci to C10 group .

[0020] Another aspect of the present invention relates to a method for treating excess bodyweight including overweight and obesity and related metabolic and cardiovascular diseases and / or a disease or a disorder associated with decreased lipolysis and / or increased adipogenesis and / or for inducing fat loss , resulting in weight loss , in a subj ect comprising the step of administering a composition as defined above to a subj ect in need thereof .

[0021] Yet another aspect of the present invention relates to a non-medical use of a composition as defined above for increasing lipolysis in a subj ect , preferably for inducing fat loss in a subj ect , wherein the at least one compound represented by Formula ( I ) is administered orally to a subj ect in an amount of 200 pg / kg body weight to 8000 pg / kg body weight . Non-medical compositions include in particular dietary supplements which are preferably administered orally to a subj ect , in particular to a human . Hence , another aspect of the present invention relates to dietary supplement comprising a composition as defined above .

[0022] Excess body weight may not only have adverse ef fects on the health of an individual or mammal but might also have an influence of the shape of the body . Hence , the compounds and extracts of the present invention can not only be used therapeutically but also cosmetically . Due to their properties to induce lipolysis , the compounds and extracts of the present invention can be contacted with tissue containing an increased amount of adipocytes leading to a reduction of adipocytes and / or the amount of fat accumulated in these cells .

[0023] Thus , the present invention relates also to a cosmetic method for improving the texture of the skin of a subj ect comprising the step of administering a composition as defined herein to said subject.

[0024] BRIEF DESCRIPTION OF THE FIGURES

[0025] Fig. 1 shows LC-PDA chromatograms of total lemon peel (A) , flavedo layer (B) and albedo layer (0) . Separation on Zorbax Eclipe Plus 018 column (1.8 pm, 100 x 2.1 mm) ; detection at UV 318 nm; 1 = limettin; 2 = bergamottin, 3 = 5-geranyloxy-7- methoxy coumarin.

[0026] Fig. 2 shows protein expression of master regulators PPARy (A) and C / EBPa (B) is downregulated by 5-geranyloxy-7-methoxy- coumarin and bergamottin in a concentration dependent manner. (0) Representative Western blots. Data represent mean values ± SEM of 3 individual experiments. *p<0.05 vs vehicle-treated cells (ANOVA with Dunnet's post hoc test) ; pre, preadipocytes; 0, citropten; B, bergamottin; G, 5-geranyloxy-7-methoxycouma- rin; DU, densitometric units.

[0027] Fig. 3 shows 5-Geranyloxy-7-methoxycoumarin and bergamottin (10 pg / ml each) did not affect protein expression of C / EBPp (isoforms LAP*, LAP, and LIP) and C / EBP5 in 3T3-L1 cells (A) . Citropten (25 pg / ml) significantly upregulated expression of LIP. Data represent mean valueslSEM of 3 individual experiments; *p<0.05 vs vehicle-treated cells (ANOVA) with Dunnet's post hoc test) . Transcriptional activities of C / EBPp and C / EBP5 were not affected by 5-geranyloxy-7-methoxy- coumarin and bergamottin (B) . Citropten significantly upregulated activity of C / EBPp. Data represent mean valueslSEM of 4 individual experiments performed in triplicate; *p<0.05 vs mock transfection (ANOVA with Dunnet's post hoc test) . (C) Assessment of transfection efficiency. C, citropten; B, bergamottin; G, 5-geranyloxy-7-methoxycoumarin; DU, densitometric units.

[0028] Fig. 4 shows effects of coumarins on expression and activity of AMPK and ACC in short-term experiments. Phosphorylation of (A) AMPK (Thrl72) and (C) ACC (Ser79) was significantly increased by bergamottin and 5-geranyloxy-7-methoxycoumarin (10 pg / ml each) . Total protein levels of AMPK (B) and ACC (D) were not affected. Data represent mean valueslSEM of 7-9 experiments; *p<0.05 vs vehicle-treated cells (ANOVA with Dunnet's post hoc test) C, citropten; B, bergamottin, G, 5- geranyloxy-7-methoxycoumarin; DU, densitometric units.

[0029] Fig. 5 shows (A) microscopic imaging of mature 3T3-L1 adipocytes (dlO; 10-fold magnification) . (B) Citropten (25 pg / ml) enhances f orskolin-stimulated lipolysis for up to 6 h. Data represent mean valueslSEM of 5-7 individual samples. (C) Concentration-dependent effect of citropten on f orskolin-stimulated lipolysis (3 h) . Data represent mean valueslSEM of 9 individual samples. *p<0,05 versus vehicle-treated control (ANOVA with Dunnet's post hoc test) . (D) Effect of citropten (25 pg / ml) on basal lipolysis (3 h) . Data represent mean valueslSEM of 5 experiments performed in duplicate or triplicate. *p<0,05 versus vehicle-treated control (Student's t-test for paired samples) ; C; citropten.

[0030] Fig. 6 shows effects of coumarin derivatives on forskolin- induced lipolysis of 3 hours (A, B) . Concentrations were 30 pM (10 pg / ml) for bergamottin (B) and 5-geranyloxy-7- methoxycoumarin (G) , 121 pM (25 pg / ml) for citropten (C) , 56 pM (10 pg / ml) for aesculetin (A) , 52 pM (10 pg / ml) for isoscopoletin (I) , 142 pM (25 pg / ml) for herniarin (H) , 81 pM (25 pg / ml) for toddalacton (Todd) , umckalin (U) and fraxinol (F) , respectively. Data represent mean valueslSEM of 6-10 individual samples. *p<0,05 versus vehicle-treated control (ANOVA with Dunnet's post hoc test) . (C) Structures of coumarin derivatives.

[0031] Fig. 7 shows the effect of citropten on diet-induced obesity in mice. Male mice (5 weeks of age) were fed a highfat diet (HFD) containing 45 kJ% fat and 35 kJ% carbohydrates for 16 weeks. After 8 weeks, animals were randomized into two groups. One group received citropten (25 mg / kg body weight (BW) ) in corn oil per os 3 x weekly for another 8 weeks. Control animals was treated with corn oil as vehicle. (A, B) Increase of body weight (BW) seems attenuated by application of citropten over the period of 8 weeks. (C, D) Abdominal white fat mass was significantly reduced by treatment with citropten. Data represent mean valueslSEM of 5 animals. (E) Schematic illustration of experimental set up. DESCRIPTION OF EMBODIMENTS

[0032] The compounds of Formula (I) of the present invention, which can also be considered as coumarin derivatives, can be synthesized chemically or can be extracted and / or isolated from natural sources. Methods for synthesizing these compounds are well known to a person skilled in the art (Ortiz-de-Elguea et al. (2021) , ACS Omega, 6(44), 29483-29494) .

[0033] The compounds of the present invention which are naturally occurring can be extracted and / or isolated from natural sources which typically comprise coumarin and derivatives thereof. Respective methods for extracting and / or isolating these compounds are well known to a person skilled in the art. Coumarin derivatives are exemplarily occurring in the peel of lemon fruits, preferably ripe lemon fruits (e.g. Citrus x limon L., Rutaceae) . Coumarin and coumarin derivatives can be extracted using extraction methods with appropriate organic and inorganic solvents— ( e . g . Stanley & Vannier (1957) , J. Am. Chem. Soo., 79 (13) , 3488-3491; Dugrand et al. (2013) , J. Agric. Food Chem., 61, 10677-10684; Jungen et al. (2021) , Food Chem. , 359, 129804) .

[0034] „Disease or a disorder associated with decreased lipolysis and / or increased adipogenesis", as used herein, refers to diseases and disorders which are caused or the result of a decreased lipolysis or increased adipogenesis.

[0035] The term "treatment", as used herein, refers to the medical management of a patient with the intent to cure, ameliorate, stabilize or prevent a disease or disorder. The term includes also palliative treatment, i.e., treatment designed for the relief of symptoms rather than the curing of the disease or disorder; preventative treatment, i.e., treatment directed to minimizing or partially or completely inhibiting the development of the associated disease or disorder; and supportive treatment, i.e., treatment employed to supplement another specific therapy directed toward the improvement of the associated disease or disorder. Thus, the term covers any treatment of a subject (human subject or human) and / or mammal including the prevention of the disease from occurring in a subject and / or mammal that can be predisposed to the disease but has not yet been diagnosed as having it ; the inhibition of the disease , i . e . , arresting its development ; and / or the relieve of the disease , i . e . , causing regression of the disease .

[0036] The at least one compound of the present invention represented by Formula ( I ) is preferably administered to a subj ect in an amount of 200 pg / kg body weight to 8000 pg / kg body weight . It turned out that a certain amount of the compound of the present invention is advantageous or even required in order to induce lipolysis in humans and / or mammals ( see examples below) .

[0037] According to a preferred embodiment of the present invention the compounds of the present invention, preferably the at least one compound of the present invention represented by Formula ( I ) , more preferably citropten and / or fraxinol , are administered to a sub ect / individual once , twice , three times or four times daily, every second day or every third day for at least 1 week, preferably for at least 2 weeks , more preferably for at least 3 weeks , more preferably for at least 4 weeks , more preferably for at least 8 weeks , more preferably for at least 6 months , more preferably for at least 12 months , more preferably for at least 24 months , more preferably for at least 36 months .

[0038] According to a preferred embodiment of the present invention R4 and R5 are independently an aliphatic Ci to C10 group, preferably an aliphatic Ci to Ce group, more preferably an aliphatic Ci to C5 group, more preferably an aliphatic Ci to C3 group .

[0039] According to another preferred embodiment of the present invention R4 and Rs are independently an alkane group, an alkene group or an alkyne group .

[0040] According to a further preferred embodiment of the present invention R4 and / or Rs are a methyl groups .

[0041] According to a particularly preferred embodiment of the present invention the at least one compound is citropten ( limettin) or fraxinol .

[0042] Citropten is preferably used as a pure substance and / or as part of an extract , preferably of a plant extract , more preferably of a lemon extract . Citropten-enriched citrus extracts or citropten as a pure substance can be used for the adjuvant treatment of lipometabolic diseases, lipid metabolism disorders, obesity and secondary diseases like type 2 diabetes and cardiovascular diseases. Fraxinol can be used for the same purpose as citropten and also as pure substance and / or as part of an extract, preferably of a plant extract, more preferably of an extract of Fraxinus excelsior, Elsholtzia bodinieri or Prunus prostrata, for instance.

[0043] In an particular preferred embodiment of the present invention citropten and fraxinol are administered in combination to a subject. The total amount of citropten and fraxinol administered may be 200 pg / kg body weight to 8000 pg / kg body weight. The weight ratio between citropten and fraxinol in the composition of the present invention may range from 1:500 and 500:1, preferably from 1:400 to 400:1, more preferably from 1:300 to 300:1 , even more preferably from 1:200 to 200:1, even more preferably from 1:100 to 100:1, even more preferably from 1:50 to 50:1, even more preferably from 1:20 to 20:1, even more preferably from 1:10 to 10:1, even more preferably from 1:5 to 5:1, even more preferably from 1:2 to 2:1. In a particularly preferred embodiment of the present invention the weight ratio between citropten and fraxinol in the composition of the present invention may be 1:1.

[0044] According to a preferred embodiment of the present invention the disease or a disorder associated with decreased lipolysis and / or increased adipogenesis is obesity.

[0045] According to another preferred embodiment of the present invention said composition comprises further coumarin or a further derivative thereof.

[0046] The composition of the present invention may comprise further active compounds, in particular coumarin or a further derivative thereof.

[0047] According to a preferred embodiment of the present invention the further coumarin derivative is selected from the group consisting of bergamottin, 5-geranyloxy-7-methoxycoumarin, herniarin, aesculetin and isoscopoletin .

[0048] 5-Geranyloxy-7-methoxycoumarin-enriched citrus extracts and 5-Geranyloxy-7-methoxycoumarin as a pure substance, bergamottin-enriched citrus extracts and bergamottin as a pure substance , aesculetin-enriched extracts and aesculetin as a pure substance , and isoscopoletin-enriched extracts and isoscopoletin as a pure substance can be used for the adj uvant treatment of lipid metabolism disorders , obesity and secondary diseases ( type 2 diabetes and cardiovascular diseases ) .

[0049] Hence , the composition may comprise at least one compound of Formula ( I ) , preferably citropten and / or fraxinol , in combination with coumarin and / or one of the aforementioned coumarin derivatives . The composition comprises preferably a compound of Formula ( I ) and coumarin, a compound of Formula ( I ) and bergamottin, a compound of Formula ( I ) and 5-geranyloxy-7- methoxycoumarin, a compound of Formula ( I ) and herniarin, a compound of Formula ( I ) and aesculetin, a compound of Formula ( I ) and isoscopoletin, a compound of Formula ( I ) , coumarin and one or more of the aforementioned coumarin derivatives .

[0050] Particularly preferred combinations comprise citropten and 5-geranyloxy-7-methoxycoumarin-enriched citrus extracts or citropten and 5-geranyloxy-7-methoxycoumarin as pure substances and can be used for the adj uvant treatment of lipid metabolism disorders , obesity and secondary diseases ( type 2 diabetes and cardiovascular diseases ) . Further preferred combinations comprise citropten and 5-geranyloxy-7-methoxycouma- rin for the adj uvant treatment of obesity and secondary diseases ( type 2 diabetes and cardiovascular diseases ) . Also , citrus extracts enriched with citropten, 5-geranyloxy-7-meth- oxycoumarin and bergamottin or citropten, 5-geranyloxy-7-meth- oxycoumarin and bergamottin as pure substances can be used in combination for the adj uvant treatment of lipid metabolism disorders , obesity and secondary diseases ( type 2 diabetes and cardiovascular diseases ) . Instead of citropten and / or citropten enriched extracts , fraxinol and / or fraxinol enriched plant extracts may be used in combination with the aforementioned coumarin derivatives for the same purpose .

[0051] According to a further preferred embodiment of the present invention the composition comprises at least one excipient and / or at least one physiologically acceptable carrier . According to another preferred embodiment of the present invention the composition is administered perorally, sublingual , percutaneously, topically, subcutaneously, rectally, vagi- nally, or intramuscularly to a subj ect in need thereof .

[0052] According to a preferred embodiment of the present invention the composition is provided in the form of an oral dosage form, a topical dosage form, a cutaneous dosage form, a trans- dermal dosage form, a sublingual and buccal dosage form, an intranasal dosage form, a rectal or vaginal dosage form .

[0053] According to a further preferred embodiment of the present invention the composition is provided in the form of a tablet , a capsule , a solution, an emulsion, a lotion, an ointment , a cream, a gel , a paste , a transdermal patch, a transdermal system, a cutaneous patch, a nasal spray and oromucosal solution, a spray, granules or drops , a suppositorium .

[0054] The composition of the present invention may include at least one excipient . Excipients are typically pharmacologically inactive substances used as a carrier for the active ingredients of the composition . Excipients can include substances that are used to bulk up formulations , allow for convenient and accurate dosage , stabili ze the active ingredients , and make the delivery system optically and / or organoleptically acceptable . Examples of pharmaceutical excipients include starch, glucose , lactose , sucrose, gelatin, malt , rice , flour, chalk, silica gel , sodium stearate , glycerol monostearate , talc, sodium chloride , dried skim milk, glycerol , propylene , glycol , water, ethanol and the like . Remington ' s Pharmaceutical Sciences , by E . W . Martin, Mack Publishing Co . , Easton, PA, 19th Edition ( 1995 ) , describes excipients and formulations suitable for pharmaceutical delivery of compositions . The use of pharmaceutically acceptable excipients does not imply that that product so made is useful only for pharmaceutical purposes . Rather it implies that the product is suitable for administration to or consumption by a subj ect or mammal , for example as a pharmaceutical or nutraceutical that is suitable for oral ingestion by a subj ect .

[0055] In general , the nature of the excipient will depend on the particular mode of administration being employed . For instance, solid compositions (like powder, pill, tablet or capsule forms) , conventional non-toxic solid vehicles can include, for example, pharmaceutical grades of mannitol, lactose, starch, or magnesium stearate.

[0056] The composition of the present invention may be provided in a lyophilized form and is preferably adapted for the desired mode of administration, including, but not limited to tablets, gel capsules, sustained-release capsules and the like for oral administration .

[0057] Oral compositions may include an inert diluent or an edible carrier (e.g. in the form of nanoparticles or liposomes) and may be compressed into tablets or enclosed in gelatin capsules. For the purpose of oral therapeutic administration, the composition of the present invention can be incorporated with excipients and used in the form of tablets, capsules or troches. Pharmaceutically compatible binding agents and adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients or compounds of a similar nature: a binder such as, but not limited to, gum tragacanth, acacia, corn starch, or gelatin; an excipient such as microcrystalline cellulose, starch, or lactose; a disintegrating agent such as, but not limited to, alginic acid and corn starch; a lubricant such as, but not limited to, magnesium stearate; a glidant, such as, but not limited to, colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; and a flavoring agent such as peppermint, methyl salicylate, or fruit flavoring .

[0058] When the dosage unit form is a capsule, it can contain, in addition to material of the above type, a liquid carrier such as a fatty oil. In addition, dosage unit forms can contain various other materials, which modify the physical form of the dosage unit, for example, coatings of sugar and other enteric agents .

[0059] Capsule shells are typically made of either animal based or plant-based components that readily dissolve or disintegrate after ingestion. Animal based components may include gelatin manufactured from the collagen of animal skin and / or bone. In certain embodiments, the capsule is made of gelatin. Other suitable matrix substances such as total synthetic polymer chemicals having gelatin-like properties may be used to manufacture the capsules. Plant-based components may include vegetable starch, cellulose, hypromellose (a polymer formulated from cellulose) or pullulan (polysaccharide polymer produced from tapioca starch) . In some embodiments, plant-based components may include carrageenan, potato starch, cassava starch, cornstarch, arrowroot or combinations thereof. The composition, manufacture and use of capsule shells are well known in the art .

[0060] The composition of the present invention is particularly preferred administered topically in the form of a topical dosage form, a cutaneous dosage form or a transdermal dosage form. Hence, the composition of the present invention is provided as a lotion, an ointment, a cream, a gel, a paste, a transdermal patch, a transdermal system and a cutaneous patch. The topical administration of the composition of the present invention allows, i.e., to reduce topical fat.

[0061] According to another preferred embodiment of the present invention the at least one compound represented by Formula (I) is administered to a subject in a therapeutically effective amount to treat said subject, preferably in an amount of 200 pg / kg body weight to 8000 pg / kg body weight, preferably 400 pg / kg body weight to 6000 pg / kg body weight, more preferably 500 pg / kg body weight to 5000 pg / kg body weight, even more preferably 800 pg / kg body weight to 4000 pg / kg body weight.

[0062] "Therapeutically effective amount" refers to an amount that is sufficient to achieve the desired result or to have an effect on an undesired condition. The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the age, body weight, general health, sex and diet of the patient; the time of administration; the route of administration; the duration of the treatment. For example, it is well within the skill of the art to start doses of a compound at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired ef fect is achieved . I f desired, the ef fective daily dose can be divided into multiple doses for purposes of administration . Consequently, single dose compositions can contain such amounts or submultiples thereof to make up the daily dose .

[0063] According to another preferred embodiment of the present invention the at least one further coumarin or a derivative thereof is administered to a subj ect in an amount of 800 pg / kg body weight to 4000 pg / kg body weight .

[0064] According to a further preferred embodiment of the present invention the composition comprises 10 to 500 mg, preferably 20 to 400 mg, more preferably 30 to 300 mg, even more preferably 50 to 250 mg of said at least one further coumarin or a derivative thereof .

[0065] Another aspect of the present invention relates to a method for treating a disease or a disorder associated with decreased lipolysis and / or increased adipogenesis comprising the step of administering a composition as defined herein to a subj ect in need thereof .

[0066] A further aspect of the present invention relates to a nonmedical use of a composition as defined herein for increasing lipolysis in a subj ect , preferably for inducing fat and / or topical fat loss in a subj ect , wherein the at least one compound represented by Formula ( I ) is preferably administered orally to a subj ect in an amount of 200 pg / kg body weight to 8000 pg / kg body weight .

[0067] Another aspect of the present invention relates to a cosmetic method for improving the texture of the skin of a subj ect comprising the step of administering a composition as defined herein to said subj ect .

[0068] According to a preferred embodiment of the present invention the composition used in the cosmetic method is topically applied to the skin to be cosmetically treated or orally applied to said subj ect .

[0069] According to a preferred embodiment of the present invention the composition used in the cosmetic method is applied to said subj ect using a patch or bandage . EXAMPLE

[0070] The present invention is further illustrated by the following example, however, without being restricted thereto. Example 1

[0071] In the present example the effects of the compounds of the present invention on an established in vitro model was examined. 3T3-L1 cells represent a well-established and widely used in vitro model to study different aspects of fat cell biology (Green & Meuth (1974) , Cell, 3 (2) , 127-133) . Experimental challenge of 3T3-L1 preadipocytes with mitotic and adi- pogenic inducers (insulin, glucocorticoids, cAMP elevating compounds, and growth hormones) induces the adipogenic process that is characterized by a sequence of temporarily confined events driven by distinct transcription factors.

[0072] Expression and activation of CCAAT / enhancer-binding protein subtypes 0 and 5 (C / EBP0 and C / EBP5) dominate the early phase of adipocyte differentiation. They play a crucial role in the induction of adipogenic factors i.e. peroxisome proliferator activated receptor y (PPARy) , C / EBPa, and sterol regulatory element binding protein 1 (SREBP-1) that are expressed in the intermediate phase of differentiation. PPARy is considered as one of the key regulators of adipogenesis and most of stimulators or suppressors of adipocyte differentiation interfere with its expression and / or activity either in a direct or indirect manner. In the late phase, these transcription factors synergistically promote the terminal events of adipogenesis, i.e. expression of a broad range of proteins and enzymes that are required for maintenance of the mature lipid-loaded adipogenic phenotype.

[0073] Under experimental fasting conditions (deprivation lipid of energy sources and growth factors) mature lipid-enriched 3T3- L1 adipocytes are forced towards lipolysis of TGs to release FFAs and glycerol. Quantification of these products allows conclusions about overall lipolytic activity of 3T3-L1 cells and its modulation by activators or inhibitors. The lipolytic process has been studied intensively within the last decades and is well-characterized nowadays. It basically comprises three consecutive steps that are driven by three lipases termed adipose triglyceride lipase (ATGL) , hormone-sensitive lipase (HSL) , and monoglyceride lipase (MGL) with each enzyme catalyzing the consecutive liberation of one molecule FFA.

[0074] In the present example the effect of the compounds of the present invention, also as part of a lemon peel extract, on adipogenesis and lipolysis was examined.

[0075] Materials and Methods

[0076] 1.1. Materials

[0077] 3T3-L1 cells (ATCC® CL-173™) were purchased from ATCC® via LGC Standards GmbH (Germany) . Human embryonic kidney cells 293 (HEK293) were kindly provided by Prof. Wolfgang Graier (Gottfried Schatz Research Center for Cell Signaling, Metabolism and Aging, Medical University Graz, Austria) . complete Protease Inhibitor™ Cocktail was from Roche Life Science (Austria) . Citropten (limettin; 5, 7-dimethoxy-2H-chromen-2-one) , berga- mottin (4- [ (2E) -3, 7-dimethylocta-2 , 6-dienoxy] furo [3, 2- g] chromen-7-one ) and 5-geranyloxy-7-methoxycoumarin (5-[ (2E)- 3 , 7-dimethylocta-2 , 6-dienoxy ] -7-methoxychromen-2-one ) were obtained from Extrasynthese (France) . Herniarin ( 7-methoxy-2H- chromen-2-one ) was obtained from Santa Cruz Biotechnology (Germany) . Isoscopoletin ( 6-hydroxy-7-methoxy-2H-chromen-2- one) and esculetin ( 6, 7-dihydroxy-2H-chromen-2-one) were purchased from Carl Roth (Germany) . Fraxinol, umckalin and toddalolactone were obtained from Phytolab (Germany) . Coumarin derivatives were dissolved in ethanol to yield 1000-fold concentrated ethanolic stock solutions. All other chemicals were purchased from Sigma, Austria.

[0078] 1.2. Preparation of extracts and fractionation

[0079] Ripe lemon fuits (Citrus x limon L., Rutaceae) either of a commercially available brand (SanLucar®, no cultivar recorded) or distinct cultivars ( ' Primof lore ' , 'Lisbon', and 'Eureka' ) were used for preparation of ethanolic peel extracts. Peels including flavedo and albedo layer were removed from the inner pulp by a household knife, for removing just the flavedo layer, a household peeler was used. Air-dried material was pulverized (M20 Universal mill, IKA, Germany) , mixed 1:10 with 95% ethanol and extracted for 30 min at 40 °C on a shaking water bath (GEL 1086, Germany) in falcon tubes. Thereafter, extracts were centrifuged at 3100 x g (Eppendorf Centrifuge 5810R, Hamburg, Germany) , supernatants collected and pellets mixed with fresh ethanol in order repeat extraction. This process was performed 3 times. Combined supernatants were concentrated to a small volume using a vacuum rotary evaporator (Btichi RE111, Flawil, Switzerland) . Concentrated extracts were transferred into a snap lid glass and finally dried under nitrogen. Separated extracts from flavedo and albedo layers were prepared in the same way.

[0080] 1.3. LC-ESI-MS analysis

[0081] Liquid chromatography coupled with mass spectrometry (LC- ESI-MS) of ethanolic lemon peel extracts was performed using a Dionex UltiMate 3000 RS System (ThermoFisher Scientific, Austria) coupled to a 3000 RS Diode Array Detector and a LTQ XL Mass Spectrometer following the method of Dugrand and coauthors (Dugrand et al., 2013, J Agric Food Chem, 61 (45) , 10677- 10684. https: / / doi.org / 10.1021 / jf402763t) . Extracts were dissolved to a concentration of 1 mg / ml and centrifuged at 2500 x g for 5 min (Eppendorf Centrifuge 5810R) at ambient temperature. Supernatants (5 pl) were injected onto an Agilent Zorbax column (Eclipse Plus C18, 1.8 pm, 100 x 2.1mm) . Elution was performed at 25°C at a flow rate of 0.25 ml / min using a mobile phase gradient of formic acid in water (0.1%; A) and acetonitrile (B) and as follows: from 90% A (v / v) at 0 min to 100% B at 15 min; maintenance of 100% B until 18 min., returning to 90% A at 18.5 min, equilibration until 23 min. UV / VIS absorbance was detected in a wavelength range of 190 nm to 500 nm, chromatograms for comparison of samples were extracted at 318 nm. Mass spectral detection was carried out in m / z ranges of 50 to 2000 amu. MS conditions were set to a source voltage of 3.5 kV (positive ESI) , capillary temperature of 300 °C, source temperature of 350 °C, sheath gas flow of 65 arbitrary units (arb) , and auxiliary gas flow of 15 arb.

[0082] 1.4. Cell culture

[0083] 3T3-L1 preadipocytes were cultured in DMEM (D5796, high glucose) supplemented with 10% FBS, penicillin, and streptomycin at 37 °C in 5% CO2 atmosphere and 80% humidity. For long-term experiments, cells were seeded onto 6-well plates and grown to confluence. Adipogenesis was induced 48 h post confluence (day 0) by addition of culture medium containing 10 pg / ml insulin, 0.4 pg / ml dexamethasone, and 500 pM 3-isobutyl-l-methylxan- thine (IBMX) . Experiments were performed in the absence and presence of increasing concentrations of ethanolic citrus extracts (total peel, flavedo and albedo layers) or commercially available citropten, bergamottin and 5-geranyloxy-7-methox- ycoumarin. Ethanol (0.1%) served as vehicle. At days 3 and 5, medium was replaced by fresh medium supplemented with 10 pg / ml and 0.2 pg / ml insulin, respectively, with and without compounds. At day 7, adipogenesis was terminated by harvest of cells. In a short-term approach, differentiation was performed for 24 h. Cells were harvested by treatment with RIPA buffer (#R0278, Sigma) containing EDTA (2 mM) and complete Protease Inhibitor Cocktail (total volume of 150 pl per well) and scraped off mechanically. Cell suspensions were transferred to Eppendorf vials, homogenized by repeated sonication, and kept on ice for 10 min. Aliquots were stored at 4 °C for measurement of triglycerides (TGs) and total cellular protein content. For Western blot experiments homogenates were stored at -80 °C.

[0084] 1.5. Quantification of cellular protein and TG content

[0085] Protein concentration was determined using the Pierce™ BCA Protein Assay Kit from ThermoFisher Scientific according to the manufacturer's instructions. Briefly, 25 pl of samples (cells lysates) and standards (bovine serum albumin; BSA) were incubated with 200 pl reagent for 30 min at 37 °C and thereafter absorbance was measured at 562 nm using a SPECTROstar® Nano microplate reader (BMG LABTECH GmbH, Germany) . Cellular TGs were measured using the Infinity™ Triglycerides Liquid Stable Reagent according to the manufacturer' s instructions using glycerol as standard. Results were expressed as nmol glycerol per mg protein.

[0086] 1.6. Lipolysis experiments

[0087] 3T3-L1 preadipocytes were seeded onto 6-well plates and cultured in DMEM (D5796; high glucose) supplemented with 10% fetal bovine serum, penicillin, and streptomycin at 37 °C in 5% CO2 atmosphere and 80% humidity. Differentiation of cells were performed for 10 days as described in 2.4. Mature adipocytes were washed twice in low glucose containing DMEM (D4947) . Lipolysis was induced by addition of the adenylyl cyclase activator forskolin (10 pM) and cells were incubated in medium containing 2% fatty acid-free BSA for up to 6 h in the absence and presence of coumarins. In basal lipolysis experiments, forskolin was excluded. Aliquots of the medium were collected and release of free fatty acids (FFAs) was measured using the NEFA Assay Kit from FUJIFILM Wako Chemicals Europe GmbH (Neuss, Germany) (ASC-ACOD method) according to the manufacturer's instructions.

[0088] 1.7. Western blot analysis

[0089] Lysates were denaturated by boiling with 5-fold Laemmli buffer for 10 min at 95 °C. Samples containing 15 pg of protein were separated by SDS-PAGE on 8-12% gels for 45 min at 180 V. Thereafter, proteins were transferred onto nitrocellulose membranes for 90 min (240 mA) . After blocking with Trisbuffered saline containing 0.1% (v / v) Tween-20 and 5% non-fat dry milk for 1 h (ambient temperature) membranes were incubated overnight at 4 °C with primary antibodies. After incubation of membranes with respective horseradish peroxidase-con- jugated IgGs immunoreactive bands were visualized using Western Bright™ ECL or Western Bright™ Quantum substrates (Biozym, Austria) and chemiluminescence was quantified with the Fusion SL Imaging System (VWR International GmbH) .

[0090] 1.8. Transcriptional activity of C / EBPP and C / EBP5

[0091] Transcriptional activities of C / EBPp and C / EBP6 were measured using the C / EBP Cignal™ Reporter Assay (CCS-001L; dual luc) from Qiagen (Germany) and the Dual-Luciferase® Reporter Assay System (E1910; Promega Corporation, Germany) as recently described (Mussbacher et al., 2019, Sci Rep, 9(1) , 15403. https: / / doi.org / 10.1038 / s41598-019-51579-x) . pcDNA-mC / EBPp (Addgene plasmid # 49198) and pcDNA-mC / EBP6 (Addgene plasmid #12559) were gifts from Jed Friedman and Peter Johnson, respectively. At 80-90% confluence, HEK293 cells were co-trans- fected with C / EBPp or C / EBP6 DNA (1 pg each) and Cignal Reporter (0,5 pg) in serum- and antibiotic-free medium using TurboFect™ Transfection Reagent (ThermoFisher Scientific) . Cells were incubated for 24 h in the absence and presence of citropten, bergamottin or 5-geranyloxy-7-methoxycoumarin . Thereafter, cells were harvested and lysed according to the manufacturer's instructions. Activities of firefly luciferase and Renilla luciferase were sequentially measured as bioluminescence using the GloMax Discover System GM3000 (Promega) as described. Luminescence of each sample was normalized to Renilla luciferase activity.

[0092] 1.9. Data analysis and statistics

[0093] Statistical analysis was performed using the KaleidaGraph® software version 4.1.3 from Synergy software (USA) . Statistical significance between two groups was analyzed using unpaired or paired t-test with equal variance. To judge for statistical significance between more than two groups, analysis of variance (ANOVA) was performed using Dunnett's post-hoc test. Data were expressed as mean valueslstandard error of the mean (SEM) .

[0094] 2. Results

[0095] 2.1. Phytochemical analysis of lemon peel extract

[0096] Ethanolic extracts of total lemon peel as well as of separated flavedo and albedo layers were analyzed by LC-PDA-ESI- MS . Representative UV chromatograms are shown in Fig. 1. In extracts of total peel and flavedo layer, the major compounds could be identified as the coumarins citropten (1, 8.83 min) , bergamottin (2, 15.20 min) and 5-geranyloxy-7-methoxycoumarin (3, 15.32 min) by comparison of retention behavior and spectral data according to Dugrand et al. (Dugrand et al., 2013) as well as authentic reference compounds. The extracts of the albedo layer showed coumarins only in low amounts.

[0097] 2.2. Effects of coumarins on key adipogenic transcription factors

[0098] Bergamottin and 5-geranyloxy-7-methoxycoumarin concentra- tion-dependently suppressed protein expression of late transcription factors PPARy (Fig. 2A) and C / EBPa (Fig. 2B) which are regarded as master regulators of the adipogenic process. By contrast, protein expression of early transcription factors C / EBPP (isoforms LAP*, LAP and LIP) and C / EBP6 were not affected by bergamottin and 5-geranyloxy-7-methoxycoumarin at 10 pg / ml each (Fig. 3A) . To test for potential effects of coumarins on transcriptional activities, dual luciferase reporter assays were performed in HEK293 cells. Co-transf ection of cells with plasmids encoding C / EBPp or C / EBP6 with a firefly luciferase construct under the control of a C / EBP response element was performed in the absence and presence of citropten (25 pg / ml) , bergamottin (10 pg / ml) and 5-geranyloxy-7-methox- ycoumarin (10 pg / ml) for 24 h. As illustrated in Fig. 3B, 5- geranyloxy-7-methoxycoumarin showed a tendency to decrease C / EBP6-dependent transcriptional activity, however, results did not reach statistical significance. Interestingly, citropten increased both protein expression and activity of C / EBP0. Overexpression of C / EBPP isoforms and C / EBP6 in HEK293 cells was confirmed by Western blot (Fig. 3C) .

[0099] 2.3. Effects of coumarins on AMPK signaling

[0100] Recently, Ko and colleagues proposed that bergamottin exerts its antiadipogenic effect via activation of AMPK (Ko et al., 2018) . Considering the similar inhibitory profile on adipocyte differentiation, we probed if 5-geranyloxy-7-methoxycoumarin comparably interferes with AMPK signalling. As shown in Fig. 4A, phosphorylation of AMPK at Thr172was more than doubled in the presence of 5-geranyloxy-7-methoxycoumarin (10 pg / ml) compared to controls. Moreover, phosphorylation of its downstream target acetyl-CoA carboxylase (ACC) at Ser79was increased about 3-fold (Fig. 4B) . Total protein levels of these targets were not affected by coumarins within 24 h (Figs. 4C and 4D) .

[0101] 2.4. Effects of coumarins on lipolysis

[0102] Taking into account the promising results obtained for adipogenesis in 3T3-L1 cells we extended our study and investigated potential effects of lemon peel extracts and purified coumarin derivatives on lipolysis of TGs . In mature 3T3-L1 cells (Fig. 5) , basal and f orskolin-stimulated lipolysis was measured by release of free fatty acids (FFAs) into the medium. Surprisingly, presence of citropten enhanced forskolin- stimulated lipolysis in a time- and concentration-dependent manner. Thus, in the presence of 25 pg / ml (» 120 pM) citropten, release of FFAs was significantly increased within 0.5 and 6 h (Fig. 5B) . Since the effect of citropten was maximal at 3-4 h after induction of lipolysis (185-190% of controls) a time span of 3 h was chosen for further experiments. In Fig. 5C, the effect of increasing citropten concentrations on for- skolin-induced lipolysis is shown. A significant effect was already observed at 10 pg / ml (» 48 pM) of the coumarin. Besides the effect on stimulated lipolysis, basal release of FFAs was also increased in the presence of citropten (Fig. 5D) . The pro-lipolytic effect was found selective for citropten and fraxinol since structural related coumarin derivatives such as bergamottin, esculetin, herniarin, isoscopoletin, umckalin and toddalolactone did not significantly enhance lipolysis (Fig. 6A, B) . The structures of these compounds are depicted in Fig. 6C.

[0103] Example 2

[0104] Based on the promising in vitro results with citropten and in view of the health economical relevance of anti-obesity preparations, the effects of citropten were tested in an in vivo mouse model of diet-induced obesity, measuring obesity- related parameters like body weight and weight of white adipose tissue. Based on these in vivo experiments citropten might be established as lead compound for new treatments of overweight or obesity.

[0105] Only a few in vivo models for citropten have been described in the literature. In a dextran sulf ate-colitis mouse model citropten showed after oral application for 7 days (10 mg / kg BW or 40 mg / kg BW) anti-inflammatory and colitis-protective effects (Lee HS et al., Molecules 27 (2022) :4633) . In a breast cancer model citropten was active at a dose of 35, 70 and 150 mg / kg BW (Prince M et al., Carcinogenesis 27 (2006) : 1204-1213) . In our in vivo study we decided to use the arithmetic mean dose between 10 - 40 mg / kg BW and hence selected a dose of 25 mg / kg BW. According to literature, the equivalent dose in humans can be calculated based on body surface area by dividing the dose in the mouse model (25 mg / kg BW, for instance) by a factor of 12.3, resulting in 2.03 mg / kg BW in humans (Nair AB, Jacob S, J Basic Clin Pharma 7 (2016) :27-31) . In the current study, in order to induce overweight and obesity, at 5 weeks of age, mice received a high-fat diet (HFD) with 45 kJ% fat und 35 kJ% carbohydrates which was maintained for 16 weeks. After 8 weeks, one group received citropten (25 mg / kg BW) in corn oil for another 8 weeks whereas the control animals were treated with corn oil as vehicle. Although the animals are known to show slower weight gain compared to a diet containing 60 kJ% fat, the HFD in our study is regarded as more similar to the "Western Life Style" with high consumption of sugar. Especially the carbohydrate and monosaccharide content is inadequately represented in an over proportionally fat enriched diet (60 kJ% fat) (Speakman JR, Int J Obes (Lond) , 43 (2019) : 1491-1492) .

[0106] 1. Materials and Methods

[0107] Male C57BL / 6 mice were purchased from Janvier Labs (Le Gen- est-Saint-Isle, France) . For diet-induced obesity (DIO) , D12451 (45 kJ% fat; 10 mm pellets, red) from ssniff Spezi- alitaten GmbH, Soest, Germany was used. Citropten was obtained from Cayman Chemical via SZABO-SCANDIC HandelsgmbH (Vienna, Austria) . Mice were housed at the local animal facility in approved conventional polycarbonate cages (Ehret, Emmerdingen, Germany) , 5 per cage, with dust-free laboratory bedding and enrichment (nesting material and rodent tunnels from Abedd GmbH, Vienna, Austria) . Before induction of obesity, animals were fed standard chow diet (Altromin 1324) from Altromin, Lage, Germany) and provided drinking water ad libitum. Animals were maintained at 23 ± 1 °C, with a relative humidity of 50- 70% and kept on a regular 12-h dark / light cycle. Animal care was in compliance with the Austrian law on experimentation with laboratory animals (last amendment, 2013) based on the European Union guidelines for the Care and the Use of Laboratory Animals (European Union Directive 2010 / 63 / EU) . The experimental protocols were approved by the Austrian Federal Ministry of Science and Research (BMWFW-2023-0.625.447 ) .

[0108] At 5 weeks of age, feeding with high-fat diet (HFD) was started and maintained for 16 weeks. After 8 weeks, animals were randomized into two groups. One group received citropten (25 mg / kg BW) in corn oil (0,05 ml) per os 3 times a week for another 8 weeks. Control animals were treated with corn oil as vehicle. During the whole intervention period, animals were monitored daily for their behavior and physical appearance. Body weight was measured weekly and food intake was monitored regularly. After 16 weeks, animals were euthanized, blood was collected and tissues (white adipose tissue, liver, heart, aorta) excised. White adipose tissue was weighted and finally serum and tissues were snap-frozen and stored at -80 °C for further biochemical analysis.

[0109] 2. Results

[0110] As shown in Fig. 7A and 7B, the time-dependent increase of body weight (BW) seems attenuated by application of citropten over the period of 8 weeks. However, due to the small number of animals per group, results did not reach statistical significance. By contrast, abdominal white fat mass was significantly reduced in the citropten group (Figure 7C) . In addition, the fat mass / body weight ratio was significantly reduced in animals treated with citropten (Figure 7D) . Data represent mean values ± SEM of 5 animals. Statistical analysis was performed with Student's unpaired T-test. * indicates p < 0.05 comparing vehicle to citropten treatment.

[0111] 3. Discussion

[0112] Our first set of in vivo experiments showed that citropten was well-tolerated at the applied dose (25 mg / kg BW; 3 times weekly, 8 weeks) . We observed a trend towards reduction of body weight and a statistically significant decrease of ab- dominal / gonadal fat mass in citropten-treated mice. Improve- ment / variation of the dosing regimen (e.g., daily oral administration via citropten-enriched chow) might lead to more pronounced effects at the same or even lower doses.

Claims

CLAIMS :1 . A composition comprising at least one compound represented by Formula ( I )( I ) or a pharmaceutically acceptable salt thereof or a plant extract comprising said at least one compound or said salt thereof for the use in the treatment of excess bodyweight including overweight and obesity and related metabolic and cardiovascular diseases , wherein Ri is -O-R4 , R2 is -H or -OH and Rs is -O-R5, wherein R4 and Rs are independently an aliphatic Ci to C10 group, wherein the at least one compound represented by Formula ( I ) is administered to a subj ect in an amount of 200 pg / kg body weight to 8000 pg / kg body weight .2 . The composition for the use according to claim 1 , wherein R4 and Rs are independently an aliphatic Ci to C10 group, preferably an aliphatic Ci to Ce group, more preferably an aliphatic Ci to Cs group, more preferably an aliphatic Ci to Cs group .3 . The composition for the use according to claim 1 or 2 , wherein R4 and Rs are independently an alkane group, an alkene group or an alkyne group .4 . The composition for the use according to any one of claims 1 to 3 , wherein R4 and / or Rs are a methyl group .5 . The composition for the use according to any one of claims 1 to 4 , wherein the at least one compound is citropten and / or f raxinol .6 . The composition for the use according to any one of claims 1 to 5 , wherein said composition comprises further coumarin ora further derivative thereof , wherein the further coumarin derivative is preferably selected from the group consisting of bergamottin, 5-geranyloxy-7-methoxycoumarin, herniarin, aes- culetin and isoscopoletin .7 . The composition for the use according to any one of claims 1 to 6 , wherein the composition comprises at least one excipient and / or at least one physiologically acceptable carrier .8 . The composition for the use according to any one of claims 1 to 7 , wherein the composition is administered perorally, sublingual , percutaneously, topically, subcutaneously, rectally, vaginally, or intramuscularly to a subj ect in need thereof .9 . The composition for the use according to any one of claims 1 to 8 , wherein the at least one compound represented by Formula ( I ) is administered to a subj ect in an amount of 400 pg / kg body weight to 6000 pg / kg body weight , preferably 500 pg / kg body weight to 5000 pg / kg body weight , more preferably 800 pg / kg body weight to 4000 pg / kg body weight .10 . The composition for the use according to any one of claims 1 to 9 , wherein the composition comprises 10 to 500 mg, preferably 20 to 400 mg, more preferably 30 to 300 mg, even more preferably 50 to 250 mg of said at least one compound represented by Formula ( I ) .11 . The composition for the use according to any one of claims 6 to 10 , wherein the at least one further coumarin or a derivative thereof is administered to a subj ect in an amount of 800 pg / kg body weight to 4000 pg / kg body weight .12 . The composition for the use according to any one of claims 6 to 11 , wherein the composition comprises 10 to 500 mg, preferably 20 to 400 mg, more preferably 30 to 300 mg, even more preferably 50 to 250 mg of said at least one further coumarin or a derivative thereof .

13. Non-medical use of a composition as defined in any one of claims 1 to 12 for increasing lipolysis in a subject, preferably for inducing fat loss in a subject, wherein the at least one compound represented by Formula (I) is administered orally to a subject in an amount of 200 pg / kg body weight to 8000 pg / kg body weight.

14. A cosmetic method for improving the texture of the skin of a subject comprising the step of administering a composition as defined in any one of claims 1 to 12 to said subject.

15. Cosmetic method according to claim 14, wherein the composition is topically applied to the skin to be cosmetically treated or orally applied to said subject.

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