Compositions for treating fat deposits in humans

Abstract: Pharmaceutical and cosmetic compositions for the treatment of superfluous fat deposits contain (a) vegetable active principles having adenylate cyclase agonistic activity or (b) vegetable active principles having antiphosphodiesterase activity or a combination of the two kinds of active principles (a) and (b).

Description: The present invention concerns compositions for treating fat deposits in humans. More particularly, the invention relates to compositions containing (a) vegetable active principles having adenylate cyclase agonistic activity optionally in combination with (b) vegetable active principles having antiphosphodiesterase activity. Obesity is a problem affecting a major and increasing part of western population. The weight reduction in obese subjects may be achieved either by pharmacological means or by selected therapeutic diets: however, in both cases, the more or less dramatic weight reduction does not always give satisfactory results from the aesthetic point of view since the fat deposits in some body areas (particulary thighs, gluteus, hips, knees) remain unchanged, causing unpleasant physical disproportion with consequent psychological distress of the patient. The problem of fat deposits effects also some part of the adult female population having normal weight. Compositions for treating fat deposits containing a mixture of essential oils from many different aromatic plants, among which Rosmary is cited, are described in FR 2576212. Compositions for treating paradontopathic complaints containing propolis extract combined with garden sage and camomille extract are disclosed in RO 80364. In most cases, the pharmaceutical or cosmetic compositions presently available for the treatment of fat deposits do not give the expected results. It has now been surprisingly found that compositions containing plant extracts or their selected constituents having agonist action on adenylate cyclase optionally in combination with compounds possessing anti-phosphodiesterase activity, accelerate and make the reduction of fatty deposits located in the above cited areas easier thus obtaining significant advantages both in the elimination of physiological aesthetic defects and in the consequence of the forced body weight reduction. According to the invention, suitable adenylate cyclase agonists are extracts of plants belonging to the Convolvulaceae family, such as Ipomoea hederacea , Ipomoea parassitica , Ipomoea batatas etc. or belonging to Labiatae family such as Salvia miltiorrhiza . As far as the antiphosphodiesterase activity is concerned, useful plants are those containing dimeric flavons derived from apigenine skeleton; among these, Ginkgo biloba , Sequoia sempervirens , Taxus baccata , Selaginella rupestris etc. can be used as a source of active material. The extracts of these plants or their particular fractions are used in a mixture in different ratios with adenylate cyclase agonists, and optionally with further active principles already used for the same purposes, such as yohimibine isomers. It is particularly preferred the use of apigenine dimers or corresponding derivatives such as amentoflavone, bilobetine, sciadopitisine, ginkgonetine etc, or flavone dimers having similar structure. It is known that amentoflavone is a powerful inhibitor of phosphodiesterase. It has now been found that the combination of these flavones in pure form or as extracts containing them with adenylate cyclase agonists can dramatically decrease fat deposits when topically applied on areas where fats accumulate. The previously cited Ipomoea or Labiatae species extracts are particularly active as adenylate cyclase agonists. The adenylate cyclase agonistic activity of Ipomoea hederacea , Ipomoea batatas , Ipomoea parassitica , Salvia miltiorrhiza is shown in Table 1. Adenylate cyclase activity of various crude extracts on bovine heart homogenates. Substances Conc. mg/100ul Specific Activity Control // 21.8 + 3.9 Ipomoea hederacea 0.2 51.3 + 4.1 0.4 58.6 + 0.8 Ipomoea parassitica 0.2 32.8 + 2.6 0.4 33.7 + 1.5 Ipomoea batatas 0.2 44.6 + 3.9 0.4 56.3 + 4.6 Salvia miltiorrh. 0.2 50.3 + 5.4 0.4 57.2 + 4.8 The determination of the agonist action of these extracts on adenylate-cyclase was determined according to the method of Kurokawa et al . (Biochim. Biophys. Acta 583,467,1979) using brain, heart homogenates or tumoral cells. Adenylate cyclase activity was assayed on the basis of formation of (3H) cyclic AMP formation from the (3H) ATP. Activity ratio was expressed as a percentage of the activity with the sample against the basal activity. As a reference substance, Forskoline, a well known adenylate-cyclase agonist, has been used. The determination of the phosphodiesterase activity was carried out according to the method of Beretz et al. (Experientia, 34,1054,1977). The metod consists in measuring (3H) products resulting from incubation of (3H)-c-AMP with the phosphodiesterase preparation and with an excess of nucleotides. An anion exchange resin was used in a batch process to remove the residual substrate. The inibition of PDE by flavonoids was studied in experiments in which the concentration of c-AMP was held constant at 1 µM. The I-50 (concentration of flavonoid resulting in a 50% inhibition of c-AMP breakdown) was calculated by interpolating 2 values of inhibition ranging from 35 to 75% against the logarithm of the dose of the flavonoid added. The I-50 values of the most important dimeric flavonoids in this test are reported in Table II. I₅₀ Substances Cyclic AMP Cyclic GMP Amentoflavone 0.65 0.58 Ginkgonetine 0.88 0.49 Ginkgo/Flav. 2.46 1.85 Papaverine 5.00 12.44 Cromoglycate Na 2000 235 Assays were performed with 1 µM substrate, I₅₀ was calculated as described above. In the treatment of undesiderable aestethical body areas the combinations of the hydroalcoholic extracts of Ipomoea hederacea with the extracts of Ginkgo biloba enriched in dimeric flavons or the extracts of Salvia miltiorrhiza with the extracts of Ginkgo biloba or Selaginella rupestris were particularly interesting. To reduce fat deposits, the treatment with the above cited extracts must be continued for a few days or weeks depending on the seriousness of the unattractiveness. The dosage of the extracts or of the pure active constituents of the plants having adenylate-cyclase agonistic or antiphosphodiesterase action or alfa-2-adrenergic receptor blocking action such as α-yohimbine, may vary between 0.1 mg to 0.2 mg; these products in form of extracts or pure consituents may be incorporated in formulations containing the usual excipients, such as those reported on the Remington's Pharmaceutical Sciences Handbook, Mack Pub. Co., N.Y., U.S.A.. Particularly preferred excipients are both natural or synthetic phospholipids which increase the absorption of the active principles through the skin. The daily posology of the active ingredients in the formulations depends on the patient's conditions and on the seriousness of the pathology and can be divided in two or more administrations per day. Normally a complete cycle of therapy takes, in combination with a proper diet, a treatment of 10 to 180 days. The active principles of the compositions of the invention are prepared by extracting the vegetable material with alcohols or aliphatic ketones either alone or in admixture with water or, when the active principles are lipophilic, with aromatic and/or aliphatic hydrocarbons or with ethyl acetate. The obtained extracts may be purified from inert or undesired substances by partitioning the active principles between an aqueous phase and selective solvents. An alternative process for the purification of said extracts comprises the absorption on resins when the extracts are obtained by acqueous extraction or with water miscible solvents. The absorbed active principles may be selectively released from the resin using solvents with different polarity or high dielectric constant. The resins used for these separations are usually those having polystyrene matrix, such as Amberlites (R) and Duolites (R) . Duolite S-730 R and Amberlite XAD2 R are preferably used. According to the latter process, the active principles of Salvia miltiorrhiza and of other plants belonging to the Labiatae family may be selectively eluted using toluene/ethyl acetate mixtures preferably in the 8:2 ratio; the active principles of Ipomoea species are preferably extracted from aqueous solutions after acidification at pH around 1.5 with n-butanol or mixture of n-butanol and toluene; the extracts obtained according to the above cited procedures can be directly incorporated in cosmetic or pharmaceutical formulations and used in human medication. As far as the anti-phosphodiesterase principles are concerned, the methods employed for their preparation are those normally used for dimeric flavons. The following examples are given by way of illustration of the present invention. EXAMPLE 1 Preparation of Ipomoea hederacea extract. 100 g of Ipomoea hederacea seeds are ground and extracted several times with 400 ml of a mixture of ethanol/water 6:4 (v/v); to eliminate drug impurities and flocculated materials, the combined extracts is filtered and then concentrated under vacuum to dryness; 14 g of brown solid were obtained which are ready for biological application. EXAMPLE 2 Preparation of Salvia miltiorrhiza purified extract. 500 g of Salvia miltiorrhiza roots are ground and extracted 5 times with 2 l of a mixture of ethanol/water 1:1; to eliminate drug impurities the combined extracts are filtered and then concentrated under vacuum to 500 ml (extract/drug ratio 1:1 v/w); the cloudy suspension is extracted four times with 500 ml of ethylacetate; the organic phase after filtration on Na₂SO₄ is concentrated under vacuum to dryness. 2 g of a red extract enriched in tanshinone like substance were obtained; this purified extract, poorly soluble in water can be incorporated in oily emulsions alone for the treatment of fatty unattractiveness. EXAMPLE 3 Preparation of dimeric flavonoids mixture from Ginkgo biloba leaves. 500 g of green Ginkgo biloba leaves are extracted with methanol until exhaustion; the solvent is concentrated to small volume and the residue diluted with 100 ml of a mixture of methanol/water 1:1, this phase is extracted twice with 100 ml of chloroform. The chloroform solution is evaporated under vacuum to dryness and the residue, solubilized in 100 ml of methanol/water 60% is extracted with n-hexane. The hexane extract is eliminated while the acqueous methanol is concentrated to dryness. This residue contains about 40% of dimeric flavonoids with amentoflavone skeleton and can be used as an active ingredient in formulations useful in fat deposits reduction. EXAMPLE 4 Preparation of a cream containing as active ingredients Salvia metiorrhiza extract, Ginkgo biloba dimeric flavonoids fraction and alfa-Yohimbine. Each 100 g gream contains: Salvia miltiorrhiza extract (example 2) 0.2 g Ginkgo biloba dimeric flavonoids (example 4) 0.2 g Alfa-Yohimbine 0.05 g Soybean Phospholipids 2.00 g Carboxyvinylpolymer (Carbomer 934) 1.2 g Sodium laurylsarcosinate 0.5 g Polysorbate 80 3 g Hydrogenated lanoline 5 g Spermaceti 5 g Polyisoprene 5 g Weat Oil 2 g Dimethylsilicone oil 0.5 g Triton 80 0.1 g Parfum 0.3 g 10% Sodium hydroxide sol 2 g Water q.s.100 g

Claims: Claims for the following Contracting States : BE, CH, DE, FR, GB, IT, LI, LU, NL, SE, AT, DK Topical pharmaceutical and cosmetic compositions containing: (a) vegetable active principles having adenylate cyclase agonistic activity optionally in combination with (b) vegetable active principles having antiphosphodiesterase activity with the proviso that said principles (a) are not from Salvia officinalis and Rosmarinus officinalis. Compositions according to claim 1 wherein the active principles of a) type comprise Ipomoea hederacea , Ipomoea parassitica , Ipomoea batatas or Salvia miltiorrhiza extracts. Compositions according to claim 1 wherein the active principles of b) type comprise Ginkgo biloba , Sequoia sempervirens , Taxus baccata , Selaginella rupestris extracts. Compositions according to claim 1 wherein the active principles of b) type comprise apigenine, amentoflavone, bilobetine, sciadopitisine, ginkgonetine. Compositions according to any one of the previous claims further containing alpha-yohimbine. Compositions according to claims 1 or 5 containing Salvia miltiorrhiza extracts. Compositions according to any one of the previous claims containing natural or synthetic phospholipid mixtures as excipients. The use of (a) vegetable active principles having adenylate cyclase agonistic activity optionally in combination with (b) vegetable active principles having antiphosphodiesterase activity with the proviso that said principles (a) are not from Salvia officinalis and Rosmarinus officinalis for the preparation of a pharmaceutical composition for treating fat deposits. A method of cosmetically treating fat deposits characterised by the use of (a) vegetable active principles having adenylate cyclase agonistic activity optionally in combination with (b) vegetable active principles having antiphosphodiesterase activity with the proviso that said principles (a) are not from Salvia Officinalis and Rosmarinus Officinalis. Claim for the following Contracting States : GR, ES A process for the preparation of topical pharmaceutical and cosmetic compositions characterized in that (a) vegetable active principles having adenylate cyclase agonistic activity optionally in combination with (b) vegetable active principle having antiphosphodiesterase activity, with the proviso that said principles (a) are not from Salvia officinalis and Rosmarinus officinalis , are formulated into suitable topical administration forms.

Inventors: BOMBARDELLI EZIO, BOMBARDELLI, EZIO
Applicants: INDENA SPA, INDENA S.P.A.
Application year: 1990
Main classification: A61K35
IPC codes: A61K31, A61K8, A61K36, A61Q19, A61P3, A61P43