EP1843745A1 - Dosage forms of active ingredients containing hydroxystilbene for treating menopausal complaints - Google Patents

Abstract

The invention relates to a method for producing drug extracts that contain hydroxystilbene from a vegetable drug and to the use of various agents for pharmaceutical and non-pharmaceutical purposes.

Description

 METHOD OF DOSING HYDROXYSTYLENE-ACTIVE AGENTS FOR THE TREATMENT OF AIR CONDITIONERS COMPLAINTS

The invention relates to novel dosage forms containing hydroxystilbene, to the production of these dosage forms and their use for the treatment of climacteric complaints of the woman, juvenile oligomenorrhoea and dysmenorrhea, primary and secondary amenorrhoea or endometritis.

BACKGROUND OF THE INVENTION

Menopause is a disorder of ovarian function, accompanied by high levels of follicle stimulating hormone (FSH) and low levels of estradiol (E2). The time to last menstruation is also referred to as "perimenopause." During this phase, most women notice cyclic irregularities lasting from 1 to 5 years, and in most Western countries, about 80% of peri- and postmenopausal women suffer from hot flashes and at About 30% of them experience hot flushes so often and frequently that their quality of life is significantly impaired, and the exact physiological cause of hot flashes is unknown (Ginsburg, Obstetrics and Gynecology Clinics of North America 1994; 21 (2): 381- Hot flushes follow release of gonadotropin-releasing hormone (GnRH) from the hypothalamus and are not associated with changes in estradiol and estrone levels, as are luteinizing hormone (LH), corticotropin, growth hormone release, and beta - Lipotrophin from the pituitary and dehydroepiandrosterone and androstenedione from the Nebenni accompanied. FSH levels can vary between normal and high (> 20 IU / I).

Hormone replacement therapy (HRT) replaces the two female sexual steroids, estrogen and progesterone. For many years, HRT has been used to treat menopausal symptoms and to prevent cardiovascular diseases and osteoporosis in peri- and postmenopausal women. It was considered effective and safe. However, this has not been the case since large randomized controlled studies have been conducted that show the opposite (Beral et al., Lancet 2002; 360: 942-944, Collaborative Group, Lancet 1997; 359: 1047-1059). HRT has no positive effect on the incidence and progression of coronary heart disease. There are even indications that they reduce the risk of cardiovascular disease from peri-and postme-

M / 45333-PCT Climacteric increased nopausal women. In a randomized, double-blind, placebo-controlled, primary estrogen plus progestin study conducted by the Women's Health Initiative (WHI), 100% more thromboses, 41% more strokes, and 29% more myocardial infarctions were found after HRT compared to placebo ( Writing Group for WHI Investigators JAMA 2002; 288: 321-333, Chlebowsky et al., JAMA 2003; 289: 3243-3253). In addition, randomized controlled trials have found that HRT is associated with an increased risk of breast cancer (Million Women Study Collaborators Lancet 2003; 362: 419-427, Schairer et al., JAMA 2000; 283: 485-491 Ross et al J Natl Cancer Inst 2000; 92: 328-332, Colditz et al., N Engl J Med 1995; 332: 1589-1593, Magnusson et al., Int J Cancer 1999; 81: 339-344).

The estrogen receptor (ER) mediates the activity of estrogens in the regulation of a number of important physiological processes, including the development and function of the female reproductive system. While the stimulation of processes in these tissues has important health benefits, stimulation of other tissues, such as the breast and uterus, especially at the peri- and postmenopausal stage, may increase the risk of cancer. The ER family includes two subtypes, ERa and ERβ. Studies with ER-selective knock-out mice have shown that the typical undesirable estrogenic effects, such as endometrial hyperplasia and breast cancer, are mediated by ERa, while ERβ acts as a negative regulator of ERα and for protection against hyperproliferation and carcinogenesis in tissues, where both receptors are coexpressed (Hewitt et al Brest Cancer Res 2003; 2: 345-352, Frasor et al Endocrinology 2003; 144: 3159-3166, Lindberg et al Mol Endocrinol 2003; 17: 203-208 ).

Based on the results of clinical and experimental studies, more and more women refuse to use HRT for their menopausal symptoms. In addition, HRT is not indicated for the treatment of women who already had an endocrine-dependent tumor or who are at high risk for cancer, especially breast cancer and endometrial cancer. Therefore, new drugs for the treatment of menopausal symptoms, which are especially free in the long-term use of the potential risks associated with HRT, very desirable.

M / 45333-PCT Climacteric There are several so-called phytoestrogens, which claim to be effective in reducing hot flashes and sweating. For most of these plant preparations, the results of clinical trials, if any, are contradictory or have no efficacy compared to paceboebo. In addition, some ingredients of these preparations may have estrogenic activity despite their relatively weak binding affinity for ERα (eg, genistein, coumestrol, equol and zearalenone; Müller et al Toxicol, 2004: 80: 14-25) and thus a risk similar to HRT represent. In fact, long-term treatment with soybean phytoestrogens has been observed to result in endometrial hyperplasia (Unfer et al., Fertil Steril 2004; 82: 145-148). These observations challenge the long-term safety of phytoestrogens, particularly with respect to the endometrium.

There is thus a strong need for new drugs or drug combinations for the treatment of climacteric complaints that do not bind to the ERa, i. are not pro-carcinogenic, and do not promote the development of cardiovascular disease.

Since 1993 in Germany under the name extract Rheum rhaponticum (ERR 731 ^® ) (trade name Phytoestrol ^® N) a dry extract of roots of Rheum rhaponticum for follicle hormone replacement therapy, for example, for the treatment of women with climacteric symptoms, juvenile oligomenorrhea and dysmenorrhea, primary and secondary amenorrhea as well as endometritis on the market. The ingredients of the specific ERr 731 ^® extract are rhaponticin, deoxyrhaponticin, rhapontigenin and deoxyrhapontigenin (Table 1).

Table 1

M / 45333-PCT Climacteric All ingredients of ERr 731 ^® belong to the group of hydroxystilbenes. Several studies have shown that the number and position of the free hydroxy and methoxy groups strongly influence the biological activity of the hydroxystilbene. The pharmacological effect of hydroxystilbene is also dependent on the presence of a glucose group.

R ¹ R ² R ³

Resveratrol OH H OH

Rhaponticin OCH ₃ OH 0-Glc

Deoxyrhaponticin OCH ₃ H 0-Glc

Rhapontigenin OCH ₃ OH OH

Deoxyrhapontigenin OCH ₃ H OH

Astringin OH OH 0-Glc

Piceatannol (Astringenin) OH OH OH

Whether or to which metabolites the ingredients of ERr 731®, e.g. after oral administration in the body are degraded, is insufficiently studied. Thus, it is known only from studies on the antithrombotic and anti-allergic activity of rhizonticin-containing extracts of Rhei rhizoma that rhapontin is degraded by bacteria of the human intestinal tract to rhapontigenin (Park et al, Arch. Pharm. Res. 2002, 25 (4), 528 -533). Metabolism of rhaponticin to piceatannol or deoxyrhaponticin to resveratrol has not previously been described.

The preparation which is available commercially ERr 731 ^® (trade name Phytoestrol® ^® N) has certain disadvantages with respect to dosage form and manufacturing. In particular, the absolute active ingredient content in the commercial product is relatively low, so that the dosing

M / 45333-PCT Climacteric form for a drug dose of, for example, 4 mg is relatively large (400 mg tablet), which is sometimes found to be detrimental in oral administration.

The size of the dosage form also has a detrimental effect on the production costs, since a disproportionately high proportion of formulation auxiliaries is contained relative to the amount of active ingredient, which in turn unnecessarily increases the production costs. A reduction of the solid dosage form is not readily achievable, since reducing the size of the dosage form makes it even more difficult to comply with the limit for the still permissible variation in the active ingredient content. This applies in particular to active compounds of the present type which tend to segregate strongly during the preparation of the solid dosage form, in particular of the active ingredient-containing tablet core.

Since in the solid dosage forms used for the above indication a release of active ingredient in the intestine is partly desired, conventional formulations are provided with a gastro-resistant coating. Such coatings usually have a softening agent to avoid undesirable cracking, which often causes incompatibilities, e.g. Allergies leads, especially with prolonged intake of the drug. On the other hand, there is a risk of triggering stomach pain when cracks occur. Crack-resistant, but unplasticized, enteric-coated solid formulations would therefore also be desirable.

Disadvantages of the currently available on the market solid oral dosage forms are sometimes seen in the fact that the ingredients can not work immediately, since the release takes place with a certain time delay and thus treatment of acute complaints, such as sudden migraine or hot flashes, not optimal. In addition, conventional solid dosage forms are only systemically and not locally administrable, resulting in localized health disorders, e.g. the occurrence of vaginal problems and no other child-related complaints is a disadvantage. In addition, solid dosage forms, in particular enteric-coated tablets have only limited individual dosage, because not divisible, which may also be disadvantageous. Alternative, appropriate dosage forms of the above active ingredients would therefore be desirable.

M / 45333-PCT Climacteric Against the provision of dosage forms, which are not designed for oral administration, such as locally applied gels, also speaks that it has previously been assumed that the resveratrol and piceatannol precursors, such as rhaponticin and deoxyrhaponticin, are per se pharmacologically inactive, and Only by metabolism after oral administration to resveratrol and Piceatannol effect show.

FR 2 835 185 describes a complex rhubarb extract obtainable from rhizomes of Rheum rhaponticum which is said to contain at least 50% hydroxystilbene, at least 50% of these hydroxystilbenes being rhaponticin, deoxyrhaponticin, astrangine and piceatannol. A preferred extract contains 15-50% by weight of rhaponticin, 10-35% by weight of deoxyrhaponticin, 5-10% by weight of astrangine and 0.1-3% by weight of piceatannol. This extract is prepared by aqueous-alcoholic extraction of rhizomes of Rheum rhaponticum, as illustrated in the examples. The total amount of rhaponticin and deoxyrhaponticin that can be achieved is only 76% by weight.

Astrangin is present in an amount of 11% by weight, piceatannol in a proportion of 3% by weight, and anthracenosides in a proportion of 0.5% by weight. In addition, this extract seems to contain about 10 wt .-% further, unspecified ingredients. It is further claimed in FR 2 835 185 that due to the synergistic effects of the various ingredients of the extract, the special extract therein has biological properties which are considerably superior to the effect of the individual hydroxystilbenes, in particular those effects which the ingredients described therein are intended to have individually. The extract described there is said to have antioxidant, anti-tumor, anti-inflammatory and estrogenic properties. In fact, FR 2 835 185 does not provide a verifiable technical teaching for the alleged pharmacological applicability, let alone the alleged synergistic effect of the complex drug extract described therein. The experimental part only describes individual formulation examples for capsules, tablets or creams. In particular, any experimental data demonstrating the alleged utility for the treatment of those diseases associated with free radicals, such as accelerated aging, cancer, atherosclerosis, wrinkles, inflammatory phenomena, and the like, are lacking. Also, the alleged suitability of a combination of the rhubarb extract described therein with a prenylflavonoid-rich hops extract for the treatment of free radical diseases and / or for the treatment of hormonal imbalances.

M / 45333-PCT Climacteric balance, such as amenorrhoea, menopause, hot flashes, etc. is not proven by any data. Moreover, it remains completely unclear which of the components actually contained in the extract (rhaponticin, deoxyrhaponticin, astrangin, piceatannol, anthracenosides and non-analyzed constituents in a proportion of 10%) contribute to the claimed pharmacological activity or, if appropriate even for the claimed synergy are mandatory. The actual disclosure of FR 2 835 185 should therefore be limited to the preparation of a specific, complex rhubarb extract by aqueous-alcoholic extraction of rhizomes of Rheum rhaponticum and the production of specific hydroxystilbene derivatives as well as the preparation of various pharmaceutical formulations. In addition, there are doubts as to whether the complex extract according to FR 2 835 185 can be satisfactorily processed into tablets satisfying the quality standards of the pharmaceutical industry.

There is therefore a need for improved dosage forms for the ERr 731 extract and other agents having similar active ingredients for more efficiently treating various climacteric disorders which no longer have one or more of the disadvantages described above.

SUMMARY OF THE INVENTION

Surprisingly, this object has been achieved by providing simpler to produce, solid dosage forms of ERR 731 ^® and comparable active ingredients and drug combinations, which may also have a higher active ingredient content. The above object has also been achieved by providing for the first time semi-solid and liquid dosage forms of the above-described active compounds and combinations thereof.

Surprisingly, the following was stated:

a) Smaller solid dosage forms with higher relative active ingredient content can be produced, which show a surprisingly low percentage variation in the active ingredient content.

M / 45333-PCT Climacteric b) Smaller solid dosage forms with higher relative drug content and enteric coating are also more crack resistant than conventional tablets and can be formulated without plasticizer in the coating, resulting in fewer side effects such as stomach problems and allergies due to lack of plasticizers.

c) Due to the surprisingly proven effectiveness of resveratrol and picantannol precursors, such as rhaponticin and deoxyrhaponticin per se, there is the possibility of providing liquid or semisolid formulations which comprise these active ingredients and can be administered locally, are better and more precisely individually dosable, make the drug bioavailable faster and show fewer side effects such as stomach problems and allergies due to the lack of supplements such as emollients. Moreover, such liquid or semi-solid formulations are particularly easy to prepare.

DESCRIPTION OF THE FIGURES

Figure 1 shows the formation of piceatannol and resveratrol in vivo in male and female dogs 24 hours after administration of 100 mg ERr 731 / kg body weight.

Figure 2 shows the estrogenic activity of 17β-oestradiol compared to resveratrol, cis- and trans-rhapontigenin, deoxyrhapontigenin, piceatannol and ERr 731 in a recombinant yeast screen, stably expressing ER-α. The concentrations (X-axis) are molar concentrations with the exception of ERr 731 ^® concentration, in ug / ml of indicated.

Figure 3 shows the estrogenic activity of 17-beta estradiol compared to resveratrol, cis and trans rhapontigenin, deoxyrhapontigenin, piceatannol and ERR 731 ^®, expressed as activity of alkaline phosphatase in Ishikawa cells transiently transfected with ER-α. The concentrations (X-axis) are molar concentrations with the exception of ERr ^® 731 - concentration in ug / ml of indicated.

Figure 4 shows the results of a pharmacokinetic study Ingredient rhaponticin of ERr 731 ^® in the blood of a test subject after oral administration ERr 731 ^®. Rha-

M / 45333-PCT Climacteric ponticin could be detected in the blood but not rhapontigenin. Likewise, its metabolite piceatannol was undetectable under the experimental conditions in the blood.

Figure 5 shows the result of experiments for the activation of the estrogen receptor-beta (ERß) through the active ingredient combination ERr 731 ^® in the human endometrial carcinoma cell line HEC-1 B (Figure 5a); the aglycones trans-rhapontigenin (FIG. 5b) and deoxyrhapontigenin (FIG. 5c) are effective only at relatively high concentrations (E2 = estradiol, RLU = relative luciferase units); * = p <0.05; * ^* = p <0.01

DETAILED DESCRIPTION OF THE INVENTION

1. Preferred articles of the invention

The invention relates to a solid dosage form comprising a drug-containing solid core with a pharmaceutically acceptable carrier and an active ingredient content of about 1 to 20 wt .-%, based on the total weight of the core, wherein the active ingredient is a hydroxystilbene-containing active ingredient or a Hydroxystilbene-containing active ingredient combination, selected from resveratrol and piceatannol prodrugs (precursors), in particular rhaponticin, deoxyrhaponticin, cis- and trans-Rhapontigenin, deoxyrhapontigenin and astringin; as well as resveratrol and piceatannol; and their stereoisomeric forms, in each case in the form of their salts or in the phenol form and functional derivatives, or combinations of these compounds.

Resveratrol and piceatannol "prodrugs" within the meaning of the invention are, in particular, substances which can be partially or completely converted in vivo, eg in humans and / or another mammal, such as, for example, dogs, resveratrol and / or piceatannol Examples of sugar-containing precursors include rhaponticin, astringin and deoxyrhaponticin Typical examples of sugar-free precursors include rhapontigenin and deoxyrhapontigenin The terms "prodrug" are glycosides or sugar-free (aglycones) natural or synthetic precursors of resveratrol or piceatannol. However, in the context of the invention, "precursors" are not to be understood as a functional restriction As evidenced by the test results described below, in particular the "precursors" according to the invention per se develop advantageous pharmacological effects.

M / 45333-PCT Climacteric Preferably, the active ingredients are substantially in the trans form. Salts are especially the alkali and alkaline earth phenolates of the above compounds which have one or more free phenolic hydroxyl groups. If several hydroxyl groups are present, these may be present partially or completely in the salt form.

The active ingredient or combination of active substances to be used according to the invention is chemically synthesized or, in particular, an ingredient which can be isolated from natural or recombinant plants or an ingredient combination which can be isolated therefrom and which contains at least one of the abovementioned hydroxystilbene compounds. The resulting plant extracts or individual components thereof can also be subjected to derivatization reactions in order to obtain so-called functional derivatives. These are, in particular, those derivatives which can be returned to the non-derivatized starting compound in the human or animal body after administration. Particular mention should be made of ethers and ester derivatives of the compounds used according to the invention. In this case, individual or all etherifiable or esterifiable groups of a molecule (in particular the phenolic and glucosidic hydroxyl groups) may be derivatized. Examples of suitable derivatives and their preparation are described, for example, in FR 2 835 185, to which reference is hereby expressly made. For example: esters of saturated or unsaturated, aliphatic or aromatic carboxylic acids having up to 25 carbon atoms, such as 1 to 25 carbon atoms, for example saturated C ₆ -C ₂₂ -fatty acids (such as, for example, saturated unbranched fatty acids selected from capronic acid, enanthic acid , Caprylic acid, pelargonic acid, capric acid, undecanoic acid, lauric acid, tridecanoic acid, myristic acid, pentadecanoic acid, palmitic acid, margaric acid,

Stearic acid, nonadecanoic acid, arachidic acid, behenic acid); or silyl ethers, wherein the silicon atom carries three identical or different, straight-chain or branched, saturated or unsaturated hydrocarbon radicals having up to 20 carbon atoms, such as, for example, C ₁ -C ₂₀ -alkyl or C ₂ -C ₂₀ -alkenyl.

Preferably, an active ingredient combination of at least two of the above-mentioned compounds is used, such as e.g. 2, 3, 4, 5, 6, 7 or 8 individual compounds, wherein the group of resveratrol precursors (in particular deoxyrhaponticin and deoxyrpontigenin) and the piceatannol precursors (in particular rhaponticin and rhapontigenin) is represented in each case by one compound.

M / 45333-PCT Climacteric In a preferred embodiment, the active ingredient or combination of active substances is obtainable from plants which are selected from natural plants and from genetically modified plants modified by breeding or recombinant plants which have a higher content of at least one of the desired ingredients compared to the corresponding unmodified plant. In particular, these plants are selected from plants of the genus Rheum spp., Astragalus spp., Cassia spp. or Picea spp. or active ingredient-containing parts of plants. Nonlimiting examples of suitable species of these genera are Rheum undulatum, Rheum palmatum, Rheum tataricum, Rheum officinale, Rheum wittrockii, Rheum altaicum, Rheum reticulatum, Astragalus complanatus Cassia garrettiana and Picea sitchensis.

It will also be appreciated by those skilled in the art that genera / species of different abundance and age (i.e., harvest at different times of the growing season) may be used, which in turn may affect the type, amount, and composition of the active ingredients and mixtures isolatable therefrom. Likewise, various plant parts, such as roots, rhizomes, leaves and / or stems, are basically useful.

Particularly advantageous is the active ingredient or combination of active ingredients obtainable from the roots and / or other parts of plants, in particular of Rheum rhaponticum.

In a further embodiment, the active ingredient combination comprises substantially rhaponticin and deoxyrhaponticin, wherein the active ingredient combination in particular substantially rhaponticin and deoxyrhaponticin in a weight ratio of about 10: 1 to 1: 10, such as. in the range of about 5: 1 to 1: 5 or 4: 1 to 1: 4 or 3: 1 to 1: 3 or 2: 1 to 1: 2 or about 1: 1.

In a specific solid dosage form, the active ingredient combination of essentially rhaponticin and deoxyrhaponticin is contained in a weight ratio of about 2: 1 to 1: 2.

Another preferred active ingredient combination may include rhaponticin and deoxyrhaponticin, especially in proportions given above, and rhapontigenin and / or deoxyrhapontigenin. The quantitative share of Rhapontige

M / 45333-PCT Climacteric nin and / or deoxyrhapontigenin in the total active ingredient content can vary over a wide range, and is for example in the range of about 0.01 to 20 wt .-%, in particular 0.1 to 5 wt .-%, based on the total active ingredient content.

Furthermore, combinations of active substances which have a hydroxystilbene

Total content, in particular a total content of deoxyrhaponticin, deoxyrhapontigenin, rhaponticin and rhapontigenin, or a total content of rhaponticin and deoxyrhaponticin of more than 90% by weight, e.g. 91 to 100 wt .-%, or 92 to 99 or 93 to 98 or 94 to 97 wt .-%, have.

In a further preferred embodiment, a drug combination is used which is substantially free of aglycone derivatives of rhaponticin and deoxyrhaponticin, in particular resveratrol and piceatannol. "Substantially free" means an aglycone content of less than 5% by weight, in particular less than 2% by weight, for example less than 1% by weight or 0.1% by weight, such as 0 to 0, 05 wt .-%, each based on the total content of rhaponticin plus deoxyrhaponticin.

In a further preferred embodiment, the active substance combination used is a dry plant extract which has a high proportion of glycosides, in particular glycosides of the type described above. Glycosides are especially the glycosidic precursors of resveratrol and piceatannol described above. These are z, B in a proportion of 30 to 100 wt .-%, 50 to 100 wt .-%, but preferably in proportions of more than 76 wt .-%, such as 76 to 99 wt .-% or 80 to 98 Wt .-% or 85 to 96 wt .-%, each based on the total weight of the dry extract.

Further preferred are such drug combinations having a content of less than 0.5% by weight, e.g. 0 - 0.49 wt.% Or 0.001 to 0.3 or 0.01 to 0.2 or 0.01 to 0.1 wt .-% of anthraquinone and / or anthraquinones (in each case based on dry weight of the active ingredient combination). Anthrachinoids are to be understood in the broadest sense as substances with anthraquinone backbone.

A "dry extract" within the meaning of the invention is in particular present when the residual moisture content, that is to say the residual proportion of water and / or organic liquid (such as extractant) is less than about 5% by weight, in particular less than 2% by weight.

M / 45333-PCT Climacteric %, such as 0 to 1.5 wt .-% or 0.1 to 0.5 wt .-%, each based on the total weight of the obtained dry extract, is.

Nonlimiting examples of suitable solid dosage forms are those wherein the active ingredient combination consists essentially of about 60-66% by weight of rhaponticin 30-35% by weight of deoxyrhaponticin 0-2% by weight of rhapontigenin; 0-2 wt .-% deoxyrhapontigenin exists.

Preferred solid dosage forms have a total drug content of about 2 to 20 mg per unit dose.

In particular, such solid dosage forms are the subject of the invention containing a sugar-free, especially mono- or disaccharide-free, e.g. Having lactose-free core.

Suitable solid dosage forms may be in the form of a pill, tablet, extrudate or granules.

Solid dosage forms in the form of a dragee, optionally with an enteric coating are particularly preferred. Preferably, such coatings are free of plasticizers, such as phthalates, e.g. Diethyl phthalate. Coating compositions which are particularly suitable for producing enteric, plasticizer-free coatings are selected from known natural and synthetic coating agents (see, for example, Voigt, Pharmazeutische Technologie, 7th Edition 1993, Ullstein Mosby, Berlin). Particularly suitable coating agents include, but are not limited to, shellac and cellulosic derivatives such as hydroxypropylmethylcellulose derivatives, e.g. Hydroxypropyl methylcellulose acetate succinate, available under the tradename AQOAT.

In particular, a solid dosage form having a total weight in the range of about 150 mg ± 20 mg, a core weight of 84 mg ± 10 mg and a drug content of about 3 to 10 mg should be mentioned.

M / 45333-PCT ^' Climacteric Furthermore, solid dosage forms are preferred, these having a uniformity of the active ingredient content (averaged over 10 or 20 randomly selected individual dosage units) not exceeding ± 5% by weight, such as ± 0.1 to 4 or + 0.5 to 3 or ± 1 to 2 wt .-%, based on the total weight of the unit dose, have (eg determined according to Ph. Eur. 5th Edition 2005 (5.0 / 2.09.06.00)).

The invention further provides a process for the preparation of a solid dosage form, comprising: a) mixing the active ingredient or combination of active ingredients with the pharmaceutically acceptable carrier; and b) solidifying the mixture to the drug core.

Preferably, the active ingredient or combination of active ingredients is dissolved or dispersed in an inert liquid and mixed with the carrier and the solvent removed during or after solidification.

Advantageously, the active ingredient or combination of active substances used according to the invention is prepared by a) providing a drug-containing part of a drug plant, optionally in comminuted form, b) adding an aqueous extractant thereto, c) subjecting the extractant to a liquid extract phase the mixture is recovered and the extraction is optionally repeated several times, and d) the extractant is removed from the liquid extract phases thus obtained.

Preferably, an extraction with an aqueous extractant is carried out at a pH of the mixture in the alkaline range.

The extracted drug plant is selected in particular from plants of the genus Rheum spp, Astragalus spp. Cassia spp or Picea spp.

M / 45333-PCT Climacteric In a preferred variant of the preparation process, the total amount of the active substance or of the active ingredient combination is mixed in portions with the pharmaceutically acceptable carrier, such as Avicel or a comparable carrier based on cellulose, in particular microcrystalline cellulose, and repeats the mixing process after each carrier addition, but at least one or twice. In particular, it is mixed with a ball mill over a period of 30 minutes to 3 hours, such as 1 to 2 hours. For example, one can use conventional laboratory ball mills for mixing as described in the examples. This gives a homogeneous and stable distribution of the active ingredient in the carrier.

According to a further process variant, the active substance core is provided with an enteric-free, preferably plasticizer-free, coating.

According to another preferred variant, the core is coated with sugar.

The invention also relates to liquid dosage forms containing an active ingredient or a combination of active substances as defined above in a proportion of about 0.1 to 20 mg / ml, such as. 0.5 to 15 or 1 to 10 or 2 to 5 mg / ml, in a solvent mixture comprising water and a pharmaceutically acceptable alcohol, in particular ethanol. Preferably, the solvent mixture is a water / ethanol mixture having an ethanol content of 10 to 50 or 20 to 40 or 25 to 35% by volume, e.g. 30% by volume. These liquid dosage forms are especially formulated as drops for oral administration.

The invention also provides semisolid dosage forms containing an active ingredient or a combination of active substances as defined above in a proportion of about 1 to 12, or 2 to 6 mg of active ingredient or combination of active ingredients (per gram of formulation) in a conventional semi-solid carrier. Suitable gelling carriers are well known and e.g. selected from swellable cellulose derivatives, such as hydroxypropylmethylcellulose, or polyacrylates, e.g. Carbopol, or gelatin. Such dosage forms may find application, for example, as vaginal gel or vaginal balls.

The invention also provides an agent comprising a solid, semi-solid or liquid dosage forms as defined above. Means in the context of the invention

M / 45333-PCT Climacteric in particular pharmaceutical agents or drugs, such as homeopathics, as well as medicinal plant preparations.

Another object of the invention relates to the use of a solid, semi-solid or liquid dosage form as defined above or prepared by any of the methods described above for the preparation of an agent as defined above for the treatment of menopausal symptoms of the woman, in particular for the treatment of menopausal symptoms in the pre - or postmenopause, as well as menopausal symptoms due to natural or therapeutically induced menopause. Climatic complaints include hot flushes, sweats, sleep disorders, irritability, physical and mental fatigue, sexual problems and urinary tract symptoms. The invention furthermore relates to a use for the treatment of juvenile oligomenorrhoea and dysmenorrhea, primary and secondary amenorrhea or endometritis.

Due to the excellent compatibility of the above-described active compounds or active ingredient combinations, the invention is also the use in the context of long-term therapy, which can be indefinitely. The daily dose to be administered this may mg or in the range of 0.1 to 20 mg 0.5 to 15, 1 to 10 or 4 to 8 mg of active ingredient or active ingredient combination, such as for example, ERr 731 ^®.

Treatment according to the invention comprises both an acute and a preventive administration of agents according to the invention.

Finally, the present invention also relates to the use of the above-described active compounds or active ingredient combinations, e.g. formulated in any of the dosage forms described above, for the selective activation of ERβ, i. Activation of ERβ without concomitant substantial or detectable activation of ERα in vivo or in vitro.

The invention therefore also relates in particular to the use of an active substance or a combination of active substances as defined above (in particular ERr731 or similar rhaponticin and / or deoxyrhaponticin-containing combinations) for the treatment of one of the diseases defined above by selective activation of ERβ, ie activation of ERβ without concomitant substantial or detectable activation of ERα. M / 45333-PCT Climacteric 2. Further specific embodiments of formulations according to the invention

a) dosage forms

The teaching of the invention is directed primarily to the preparation of solid, semi-solid or liquid dosage forms for the treatment of a human. Thus, the active compounds to be used according to the invention are usually administered in the form of pharmaceutical compositions comprising a pharmaceutically acceptable excipient with at least one active ingredient according to the invention and optionally further active ingredients. These compositions are especially administrable by the oral route.

Examples of suitable pharmaceutical formulations are, in particular, solid pharmaceutical forms, such as granules, tablets, lozenges, sachets, cachets, dragees and capsules, such as hard and soft gelatin capsules, globules, suppositories or vaginal dosage forms, semi-solid dosage forms, such as ointments, creams, hydrogels, pastes or Plasters and liquid dosage forms, such as solutions, emulsions, in particular oil-in-water emulsions, suspensions, for example lotions, injections and infusion preparations, eye and ear drops, nose drops, nasal spray and tinctures. Implanted delivery devices may also be used to deliver inhibitors of the invention. Furthermore, liposomes, microspheres or polymer matrices can also be used.

In preparing the compositions, active ingredients of the invention are usually mixed or diluted with an excipient. Excipients may be solid or semi-solid materials which serve as a vehicle, carrier, adsorbent or medium for the active ingredient or drug combinations.

Suitable excipients include, unless otherwise specified, lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose and methyl cellulose. Furthermore, the formulations may contain conventional pharmaceutically acceptable excipients such as lubricants, for example, tallow, magnesium stearate and mineral oil; Wetting agents; emulsifying and suspending

M / 45333-PCT Climacteric Medium; preservatives such as methyl and propyl hydroxybenzoates; antioxidants; Antiirritatives; chelating agents; coating aids; Emulsion stabilizers, film formers; gelling agents; Odor masking agents; masking flavors; resins; Hydrocolloids; Solvents; Solubilizing agents; Neutralizing agents; permeation; Pigments; quaternary ammonium compounds; Refatting and superfatting agents; Ointment, cream or oil bases; Silicone derivatives; spreading aids; stabilizers; Sterilanzien; Tablet excipients such as binders, fillers, lubricants, disintegrants or coatings; Propellant; Desiccant; Opacifiers; Thickener; waxes; plasticizers; include. A related embodiment is based on expert knowledge, such as in Fiedler, HP, Lexicon of excipients for pharmacy, cosmetics and related fields, 4th edition, Aulendorf: ECV Editi Kantor Verlag, 1996, is shown. See also Hager's Handbook of Pharmaceutical Practice, Springer Verlag, Heidelberg.

Solvents suitable for the preparation of formulations according to the invention are, in particular, monohydric or polyhydric alcohols, in particular ethanol, glycerol and mixtures thereof with water, such as e.g. 1 to 50% by volume of ethanol in water.

The preparation of dosage forms or pharmaceutical agents according to the invention is carried out using generally known methods of pharmaceutical technology, such as e.g. described in Voigt, Pharmaceutical Technology, 7th Edition 1993, Ullstein Mosby, Berlin.

In a preferred embodiment, a pharmaceutical agent is provided which comprises a solid dosage form. This solid dosage form comprises in turn a drug-containing solid core with a pharmaceutically acceptable carrier and an active ingredient content of about 1 to 20 wt .-%, based on the total weight of the core, wherein the active ingredient is a hydroxystilbene-containing active ingredient or a hydroxystilbene-containing active ingredient combination which is a compound selected from resveratrol and piceatannol prodrugs such as rhaponticin, deoxyrhaponticin, rhapontigenin, deoxyrhapontigenin and astringin; as well as resveratrol and piceatannol; and their stereoisomeric forms, in each case in the form of their salts or in the phenol form, or combinations of these compounds. Preferred drug combinations are as defined above.

M / 45333-PCT Climacteric This solid dosage form has, for example, a total drug content of about 1 to 20 mg, such as. B. 2 to 10 mg, per unit dose and may be in the form of a pill, a tablet, an extrudate or granules and, for example, be coated. If desired, it may also have an enteric coating.

The solid dosage form is e.g. prepared by mixing the active ingredient or combination of active ingredients with the pharmaceutically acceptable carrier and solidifying the mixture to the active ingredient core. This dissolves or disperses the active ingredient or drug combination in an inert liquid, mixes him / her with the carrier and removes the solvent during or after solidification. If appropriate, the drug core can then be provided with an enteric coating before the sugar is coated in the usual way.

Liquid dosage forms according to the invention are prepared by, for example, by mixing the active ingredient or ingredients, such as a ERr-731 ^® dry extract, in a suitable solvent such as. For example, a water / alcohol mixture, optionally together with other conventional additives dissolves.

Inventive semi-solid dosage forms, such as gels are produced for example by mixing the active ingredient or ingredients, such as a ERr 731 ^® dry extract, in a suitable solvent such as a water / alcohol mixture, alcohol or glycerin dissolves and the solution in the already swollen gel former, if necessary, incorporated together with other common additives.

The type and duration of administration of the medicaments according to the invention are the decision of the attending physician. Depending on the chosen route of administration, on the efficacy of the specific active compound composition, on the type and severity of the disease to be treated, on the patient's condition and on his response to the therapy, this may determine a suitable dose and a corresponding dosing schedule. For example, a suitable single dose may be about 0.1 to 50 mg, e.g. 2 to 12 mg, drug or drug combination as defined above and administered 1 to 3 times daily until the desired treatment success is observed.

M / 45333-PCT Climacteric b) Preparation of a drug extract usable according to the invention

Drug extracts which can be used according to the invention are preferably prepared by

a) provides a hydroxystilbene-containing part of a drug plant, optionally in comminuted form, b) adding to it an aqueous, organic or aqueous-organic extractant, c) after the action of the extractant, extracting a liquid extract phase from the mixture and optionally repeated the extraction several times , and d) removing the extractant from the liquid extract phases thus obtained.

In particular, the extract thus obtained comprises at least one compound selected from rhaponticin, deoxyrhaponticin, rhapontigenin, deoxyrhapontigenin in salt or in phenolic form, in a stereoisomeric form thereof, such as cis or trans form, or as a mixture of such stereoisomeric forms.

Preferably, however, the extracted hydroxystilbenes are substantially in the trans form. Salts are especially the alkali and alkaline earth phenolates of the above compounds which have one or more free phenolic hydroxyl groups. If there are several hydroxyl groups, these may be present partially or completely in the salt form.

As already mentioned, the resulting plant extracts or individual components thereof can also be subjected to derivatization reactions in order to obtain so-called functional derivatives.

Preferably, a combination of active ingredients is obtained from at least two of the above compounds, e.g. 2, 3, 4, 5, 6, 7 or 8 individual compounds, wherein the group of resveratrol precursors (especially deoxyrhaponticin and deoxyrhapontigenin) and the piceatannol precursors (especially rhaponticin and rhapontigenin) is represented by one compound each

M / 45333-PCT Climacteric In a further preferred embodiment of the method according to the invention, an extract is provided which has a high proportion of glycosides, in particular glycosides of the type described above, such as, for example, a proportion of 30 to 100% by weight, 50 to 100% by weight, 60 to 99 wt .-% or 80 to 98 wt .-% or 85 to 96 wt .-%, each based on the total weight of the obtained dry extract. A "dry extract" in the sense of the invention is in particular present when the residual moisture content, ie the residual proportion of water and / or organic liquid (such as extraction agent) is less than about 5% by weight, in particular less than 2% by weight, such as 0 to 1, 5 wt .-% or 0.1 to 0.5 wt .-%, each based on the total weight of the obtained dry extract, is.

In a further preferred embodiment, an extract is provided which is substantially free of aglycone derivatives of rhaponticin and deoxyrhaponticin, in particular resveratrol and piceatannol. "Substantially free" means an aglycone content of less than 5% by weight, in particular less than 2% by weight, for example less than 1% by weight or 0.1% by weight, such as 0 to 0, 05 wt .-%, each based on the total content of rhaponticin and deoxyrhaponticin.

Furthermore, active ingredient combinations are preferably prepared which have a total hydroxystilbene content of more than 90% by weight, such as 91 to 100 wt .-%, or 92 to 99 or 93 to 98 or 94 to 97 wt .-%, have.

Furthermore, such drug combinations are preferably prepared that have a content of less than 0.5 wt .-%, such as. 0 - 0.49 wt.% Or 0.001 to 0.3 or 0.01 to 0.2 or 0.01 to 0.1 wt .-% of anthraquinone and / or anthraquinones (in each case based on dry weight of the active ingredient combination). Anthrachinoids are to be understood in the broadest sense as substances with anthraquinone skeleton.

In a preferred embodiment, the drug plant to be extracted is selected from natural plants as well as genetically modified plants modified by breeding or recombinant, plants which have a higher content of at least one of the desired ingredients compared to the corresponding unmodified plant. In particular, these plants are selected from plants of the genus Rheum spp., Astragalus spp., Cassia spp. or Picea spp. or active ingredient

M / 45333-PCT Climacteric gene parts of plants. Nonlimiting examples of suitable species of these genera are Rheum undulatum, Rheum palmatum, Rheum tataricum, Rheum officinale, Rheum wittrockii, Rheum altaicum, Rheum reticulatum, Astragalus complanatus, Cassia garrettiana and Picea sitchensis. Preference is also the use of varietal drug plants.

It will also be appreciated by those skilled in the art that genera / species of different abundance and age (i.e., harvest at different times of the growing season) may be used, which in turn may affect the type, amount, and composition of the hydroxystilbene and mixtures isolatable therefrom. Likewise, various plant parts, such as roots, rhizomes, leaves and / or stems, are basically useful.

The particular plant part or mixture of plant parts may, if appropriate, be mechanically treated prior to extraction, e.g. ground, chopped, shredded, tamped or cut. If appropriate, predrying, e.g. 2 hours to 2 days at 30 to 50 ° C to reduce the liquid content.

In particular, the hydroxystilbene-containing part of the drug plant used for extraction is the root of the drug plant, such as e.g. of Rheum rhaponticum.

The invention particularly relates to a process in which a hydroxystilbene-containing percolate is prepared from the drug. By "percolation" is meant a continuous extraction of soluble substances from a drug by continuous renewal of the solvent, which results in a constant concentration gradient, so that a large part of all soluble substances passes into the extract.

Alternatively, continuous or periodic mixing of the batch, such as e.g. by stirring or shaking, possible.

The temperature during the extraction according to the invention is usually in the range of 10 to 5O ⁰ C, such as 25 to 35 ° C. The pressure is usually at normal pressure. If an acceleration of the extraction rate or extract quality

M / 45333-PCT Cliraacteric can be achieved, the pressure during the extraction can also be varied, such as increased or decreased.

The extraction time may vary depending on the conditions chosen, such as the type of drug, batch size, extractant used and temperature, 1 hour to several days, e.g. Require 10 to 72 hours.

If necessary, the extraction process can be repeated several times in order to ensure complete isolation, in particular of the desired ingredients. The weight ratio of drug delivered to liquid extractant can vary over a wide range as well as from extraction step to extraction step. Typically, the weight ratio of drug to extractant ranges from 10: 1 to about 1: 200, or about 1: 2 to 1:50, or 1: 4 to 1:10.

According to a variant of the method, an extraction with an aqueous, substantially free of organic solvent extractant, such as in particular water, preferably purified water, at a pH of the mixture in the alkaline range, wherein the pH of the mixture in particular in the range of about 11 to 12, such as is about 11, 3 to 11, 8.

The pH of the mixture is determined, for example, by means of an inorganic base selected from alkali and alkaline earth metal hydroxides, e.g. Calcium hydroxide or calcium oxide. For this purpose, for example, a concentrated quenching solution can be prepared by dissolving 3 to 8 parts of CaO in 20 parts of purified water. This solution is highly alkaline and has a pH in the range of about 12 to 13, e.g. from about 12.4 to 12.6, on.

The ratio of drug delivered to base, e.g. Calcium hydroxide (calculated as calcium oxide) may be in the range of about 5: 1 to 20: 1, such as 8: 1 to 12: 1 or 9: 1 to 11: 1.

Preferably, the process is carried out by precipitating the desired hydroxystilbene from the obtained alkaline liquid extract phase, for example by bringing the pH of the extract to a value in the range of about 3 to 4, such as

M / 45333-PCT Climacteric For example, 3.2 to 3.8 or 3.4-3.6, and optionally then the precipitate separated, optionally washed and optionally dried.

For acidification, any inorganic or organic acid, e.g. Hydrochloric acid or sulfuric acid, but especially organic acids such as formic acid or acetic acid.

Before separating the precipitate, it may be expedient to let the mixture rest for a few hours or days in order to achieve the most quantitative possible precipitation of the desired extracted ingredients.

The washing of the precipitate may e.g. done with purified water and is used in particular for the removal of residual acid.

By drying, for example at 30 to 50 ^c C or 35 to 45 ° C, for example, over a period of 1 to 100 hours, residual liquid is removed from the extract until the residual moisture in the range specified above. The drying takes place in a conventional manner, for example in a drying oven. Freeze-drying is also possible.

The invention will now be further elucidated by the following nonlimiting examples.

EXPERIMENTAL PART

General methods:

Determination of stilbenes by high pressure liquid chromatography (HPLC) in dry extract from rhapontic rhubarb root

a) Sample preparation:

50 mg of extract are mixed with 40 ml of a mixture of acetone and water (1: 1) in a brown glass vessel, treated for 15 minutes in an ultrasound bath and made up to 50 ml with the solvent mixture and then diluted 1:10 with the solvent mixture.

M / 45333-PCT Climacteric b) Carrying out the chromatography:

With a part of the solution obtained above, a high pressure liquid chromatography (HPLC) is performed with the following system parameters:

Sample loop: 20μl

Column: Lichrospher 5 μ RP 18, 250 x 4 mm

Guard column: Lichrospher 5 μ RP 18, 5 x 4 mm

Column temperature: 25 ^° C.

Eluent A: acetonitrile / dist. Water / phosphoric acid 85%,

15/85 / 0.05 (parts by volume)

Plasticizer B: acetonitrile / dist. Water / phosphoric acid 85%

80/20 / 0.05 (parts by volume)

Flow: 1, 5 ml / min

Column rinse: 15 min with eluent 50% B; Equilibration time 15 min

Detection: 310 nm

HPLC: Kontron Kroma 2000

Gradient:

Time% B

O ₁ O 0 0.5 0 7.5 75 8.5 100 9.5 0 12.5 0

Under the system conditions given above, the following retention times result:

Rhaponticin: approx. 5,5 min

Desoxyrhaponticin: approx. 6,8 min Rhapontigenin: approx. 7,2 min

Deoxyrhapontigenin: approx. 9.0 min

M / 45333-PCT Climacteric For a quantitative determination, the respective peak areas are determined and compared with the corresponding peak areas of a standard extract of known composition.

Production Example 1: Preparation of the dry extract ERr 731 from rhapodrubber root with an aqueous calcium hydroxide solution

For the preparation of a dry extract of rhapontic rhubarb root are used:

Drug (Radix rheum rhaponticum) 50.0 kg

Calcium oxide 5.0 kg

Purified water 190.0 kg

Acetic acid (as needed to adjust to the required pH)

The recoverable yield is between 2 and 3 kg per 50 kg of drug.

The production takes place in the following steps:

a) 5 kg of calcium oxide are first filled into a plastic tub and slurried with 20 kg of purified water. The formation of calcium hydroxide (slaked lime) taking place under these conditions leads to a strong increase in the temperature of the solution. Therefore, the calcium hydroxide can be used only after cooling. The temperature of the solution is then at 30 ⁰ C to 35 ° C.

b) 50 kg of drug are filled into a mixer and mixed with the above-mentioned slaked lime. To remove the slaked lime as completely as possible from the plastic tub, this is rinsed with 10 kg of purified water. This washing liquid is also added to the mixer.

c) The homogeneously mixed with slaked lime drug is placed in a percolator and covered with 160 kg of purified water. The percolator remains closed for 48 hours. Subsequently, the percolate is collected in a suitable vessel at a flow rate of 50 ml / min. The percolation is continued until no more percolate escapes. The drug mass is not squeezed out after termination, but discarded.

M / 45333-PCT Climacteric d) Concentrated acetic acid is added to the percolate under permanent control until a pH in the range of 3.4 to 3.6 is reached. In order to achieve the most complete precipitation of the extract, the batch rests for 5 days.

e) The extraction of the dry extract is carried out by filtration through Büchner funnel under applied vacuum. Finally, the extract is washed with 10 to 20 kg of purified water.

f) The dry extract obtained after filtration is dried in a drying oven at 40 ° C until a residual moisture tolerance of max. 1% is reached.

Rhaponticin is well soluble in aqueous solutions with an alkaline pH range, while it is precipitated in the acidic pH range (pH 3.4 - 3.6) as a yellowish substance. This is useful for its isolation. Since the root, in addition to other organic acids, above all has a high content of oxalic acid (2/3 in water-soluble and 1/3 in bound form), it must be neutralized during the isolation in order to prevent drifting of the pH into the acidic region and thus to prevent an early precipitation of Rhaponticins. This is achieved through the use of calcium oxide. This is used as slaked lime solution with a pH of 12.4 - 12.6.

If the slaked lime is homogeneously mixed with the drug, the pH of the mixture changes. It is then in the range of 11, 3 to 11, 8, which prevents precipitation of Rhaponticin, since the phenolic form has been converted into a phenolate form. Despite the high content of oxalic acid, the pH can be kept in the alkaline range. This is because the calcium hydroxide reacts with oxalic acid to form insoluble and non-toxic calcium oxalate.

Subsequent percolation with purified water extracts rhaponticin from the root. After completion of the percolation, a pH of 3.4 to 3.6 is set by the addition of acetic acid. This pH shift from the alkaline to the acidic region leads to the precipitation of rhaponticin by recycling into the phenolic form. To achieve the most complete precipitation of Rhaponticins leaves

M / 45333-PCT Climacteric you stand the approach for 5 days. Thereafter, it is filtered off. Rhaponticin remains as a yellowish substance on the filter.

The above statements on rhaponticin apply correspondingly to the other hydroxystilbene active ingredients isolated according to the invention.

Production Example 2: Preparation of a dry extract of Rhapontikrhabarb root with various organic solvents

In the Rhapontikrhabarberwurzel used here as a drug, the mainly detectable ingredients belong to the group of hydroxystilbene. Rhaponticin (Rh) is found in the roots with a share of about 6% and desoxyrhaponticin (DRh) with a share of about 4%.

When the solvent systems listed below are used in 100 times the amount, at room temperature for 10 minutes with shaking or stirring, the following summarized proportions can be extracted:

Ethanol 86% Rh 100.8%

DRh 99.5%

EthanoM5% Rh 77.1%

DRh 75.5%

Acetone Rh 88.3%

DRh 96.6%

Water, alkaline Rh 75.5%

(pH 11, DRh 60.5% adjusted with CaO solution)

No useful results were achieved with heptane.

The respective yields of crude extracts in mass fractions (based on the drug used) are the following:

M / 45333-PCT Climacteric Ethanol 86% 35.5%

EthanoM5% 32.2%

Acetone 21, 4%

Heptane 0% water, alkaline 4.5%

The extraction of rhapontic rhubarb root with ethanol-water mixtures leads to an extract which, in addition to the main ingredients rhaponticin (about 30%) and desoxyrhaponticin (about 22%), another currently. contains not yet explored stilbene in about 20% share of the extract. In addition, the aglycones Rhapontigenin (about 8%) and deoxyrhapontigenin (about 2%) and 9 more compounds, which add up to about 20%.

The extraction with acetone gives basically the same results.

The extraction with alkalized water (see conditions according to Preparation Example 1) results in an extract of higher purity.

The main ingredients rhaponticin and deoxyrhaponticin are contained in an approximately 97% proportion in the dry extract. Rhapontigenin and deoxyrhapontigenin together make up 1.1% of the extract, while stilbene, which is still unexplored, contains only 0.2%. Another 3 compounds are contained in a 2.5% proportion.

Formulation Example 1: Preparation of a solid dosage form minitablet

1. Preparation of the tablet core:

A solid tablet core is prepared using the following active ingredients and auxiliaries in the proportions indicated (Tl = parts by weight). The ingredients are mixed and tableted in three different ways:

a) Formulation for tablet core:

Purified dry extract according to

M / 45333-PCT Climacteric Preparation Example 1 from Rhapontic Rhubarb Root (ERR 731 ^®) 3.6 Tl

Microcrystalline cellulose (e.g., Avicel 57, 0 Tl (± 40%)

Sorbitol 8,, 0 Tl

Talc 2.5 Tl

Makrogol 6000 (polyglycol) 1,, 6 Tl

Povidone (K-value of about 25, such as Kollidon ^® 25) 1, 6 Tl

Sodium dodecyl sulfate (for example Texapon ^® K 12) 0.5 Tl

Magnesium Stearate (vegetable) 0.8 Tl

75.6 parts (± 40%)

By varying the amount of ERr 731® weighed and / or varying the amount of microcrystalline cellulose, any ERr 731® contents of the crude core can be obtained (such as 2, 4, 6, 8, 10, 12 mg per tablet).

b) mixing of drug and carrier

- mixture variant a:

1, 2 Tl ERr 731® be proportionally triturated with Avicel ^® in the ball mill, followed by the remaining excipients is mixed as described below, and tableted after addition.

- mixture variant b:

ERr 731® (1 g / l solvent) is dissolved in a suitable solvent (eg ethanol / water mixture 86% v / v ethanol) and mounted on Avicel ^®, dried (at 4O ⁰ C for at least 48 hours) and after the addition of the remaining Adjuvants mixed and tableted as described below.

- mixture variant c:

The entire Avicel® amount is divided into three equal portions. The first portion is spiked with the total ERr731® and placed in a laboratory ball mill (e.g.

M / 45333-PCT Climacteric 1-25 LK ₁ Alpine, Augsburg) for at least 120 minutes. Then add the second portion of Avicel®, rub again for at least 120 minutes in the laboratory ball mill. After adding the third portion of Avicel®, briefly mix again. Subsequently, after addition of the remaining auxiliaries, mixing and tabletting are carried out as described below.

Surprisingly, with this mixture variant, it is possible to significantly reduce the tendency to separate and, even with small dosage units, a very uniform active substance content of not more than ± 5% by weight (determined according to Ph. Eur. 5th edition 2005 (5.0 / 2.09.06.00)) adjust.

c) tableting:

The mixture of Avicel® and active ingredient is sieved through a screening machine (sieve diameter 1.2 mm) into a suitable mixing container and, after addition of the indicated tabletting aids (without magnesium stearate), in a suitable mixer (eg Rhönrad mixer type Standard RR M 200, Engelsmann AG / Ludwigshafen) mixed for at least 30 minutes. After adding magnesium stearate, mix again for at least 5 minutes.

The pressing is carried out on a suitable tablet press (for example rotary type Perfecta Fette 2000, Fa. Fette / Schwarzenbeck): Core weight: 84 mg ± 4.2 mg maximum deviation punch: 7 mm diameter, drageegewölbt

The ERr-731 content per core is approximately 4 mg ± 5%.

2. Preparation of the enteric coated tablet

After dedusting the tablet cores with Eudragit, an enteric coating of cellulose acetate phthalate and diethyl phthalate, dissolved in isopropanol and ethyl acetate, is applied to the tablet cores by means of a coating system.

Macrogol is dissolved in purified water. The constituents sugar (sucrose or isomalt), calcium carbonate, talc, titanium dioxide and the two povidone

M / 45333-PCT Climacteric mixed and stirred into the liquid. The suspension is stirred for 20 minutes in a jet mixer (eg Rototron type RTA 70-50).

The coating suspension is applied to the isolated cores using a panning machine. The process is repeated until an average weight of 150 mg per replenished core is reached. Finally, apply the polishing wax and then let it roll to a high gloss.

Final weight of the enteric coated tablet: 150 mg ± 7.5 mg maximum deviation.

In this way, two different tablet forms - a sugar-containing and a sugar-free - produced, with the respective excipients were used in the following parts by weight.

a) enteric-coated mini-tablets - sugary - with plasticizer in the coating

Excipients:

Coating: Eudragit L12.5% dry matter 1.350 kg (± 40%)

Diethyl phthalate 1, 749 kg

Cellulose acetate phthalate 7.770 kg

Isopropyl alcohol 75,600 kg

Ethyl acetate 77.600 kg

Talcum 2,000 kg

Coating: talc 7,182 kg

Sugar 28.747 kg

Calcium carbonate 6,410 kg

Titanium dioxide E 171 0.635 kg

povidone

(K-value about 25, for example Kollidon ^ '25) 0.756 kg

Povidone (K value: approx. 90) 0.332 kg

Macrogol 35,000 0.635 kg

Water 10,500 kg

M / 45333-PCT - Climacteric Polish: 95% carnauba wax, 5% bleached wax (eg Capol 1295 PH) 0.108 kg

b) enteric-coated mini-tablets - sugar-free - with plasticizer in the coating

Excipients:

Coating: Eudragit L12.5% dry substance 1, 350 kg (± 40%) diethyl phthalate 1.749 kg

Cellulose acetate phthalate 7.770 kg

Isopropyl alcohol 75,600 kg

Ethyl acetate 77.600 kg

Coating: talc 7,482 kg

Sorbitol and / or isomalt 28.747 kg

Calcium carbonate 6,410 kg

Titanium dioxide E 171 0.635 kg

povidone

(K value about 25, for example Kollidon ^> 25) 0.756 kg

Povidone (K value: approx. 90) 0.332 kg

Macrogol 35,000 0.635 kg

Water 10,500 kg

Polish: 95% carnauba wax,

5% bleached wax

(e.g., Capol 1295 PH) 0.108 kg

Formulation Example 2: Preparation of a Solid Dosage Form - Minitablet sugary without plasticizer

a) Preparation of the tablet core

M / 45333-PCT Climacteric The preparation is carried out analogously to Formulation Example 1.

a} Preparation of the enteric coated tablet

The preparation is carried out analogously to Formulation Example 1, but using shellac (variant A) or acrylate (variant B) instead of celulose acetate phthalate / diethyl phthalate (plasticizer).

a) Variant A.

Supporting materials: kg (± 40%)

Coating: Eudragit L12.5% dry matter 0.400 CAPOL 5270 PH 8%

(Shellac solution) 60,000 = 4,8 kg dry matter (shellac)

Isopropyl alcohol 4,000

Ethanol 96% 3,200

Talc ..2,000

Coating: talc 7,182

Sugar 28,747

Calcium carbonate 6,410

Titanium Dioxide E 171 0.635 Polyvidone

(K value of about 25, eg Kollidon ^® 25) 0,756

Povidone (K value: approx. 90) 0.332

Macrogol 35,000 0.635

Water 10,500

Polish: 95% carnauba wax 5% bleached wax

(e.g., Capol 1295 PH) 0.108

a) Variant B

Excipients: kg (± 40%)

Coating: Eudragit L12.5% dry matter 0.400 Aqoat

M / 45333-PCT Climacteric Hydroxypropylmethylcellulose acetate succinate 5.420

Distilled water 12,500

Isopropyl alcohol 4,000

Ethanol 86% 55,000

Coating: talc 9,182

Sugar 28,747

Calcium carbonate 6,410

Titanium oxide E 171 0.635

povidone

(K value about 25, eg Kollidon ^® 25) 0,756

Povidone (K value: approx. 90) 0.332

Macrogol 35,000 0.635

Water 10,500

Polish: 95% carnauba wax

5% bleached wax

(e.g., Capol 1295 PH) 0.108

Formulation Example 3: Preparation of a Solid Dosage Form - Minitablet Sugar-Free Without Plasticizer

a) Preparation of the tablet core

The preparation is carried out analogously to Formulation Example 1, but Isomalt was used instead of Avicel.

a) Preparation of the enteric coated tablet

The preparation is carried out analogously to Formulation Example 2, but using isomalt instead of sugar.

a) Variant A.

Supporting materials: kg (± 40%)

Coating: Eudragit L.12.5% dry matter 0.400 CAPOL 5270 PH 8%

M / 45333-PCT Climacteric (Shellac solution) 60,000

= 4.8 kg dry matter

(Shellac)

Isopropyl alcohol 4,000

Ethanol 96% 3,200

Talcum 2,000

Coating: talc 7,182

Isomalt 28,747

Calcium carbonate 6,410

Titanium oxide E 171 0.635

povidone

(K value of about 25, eg Kollidon ^® 25) 0,756

Povidone (K value: approx. 90) 0.332

Macrogol 35,000 0.635

Water 10,500

Polish: 95% carnauba wax

5% bleached wax

(e.g., Capol 1295 PH) 0.108

b) Variant B

Excipients: kg (± 40%)

Coating: Eudragit L.12.5% dry matter 0.400

Aqoat 5,420

Distilled water 12,500

Isopropyl alcohol 4,000

Ethanol 86% 55,000

Talc ..2,000

Coating: talc 7,182

Isomalt 28,747

Calcium carbonate 6,410

Titanium oxide E 171 0.635

povidone

(K value of about 25, eg Kollidon ^® 25) 0,756

Povidone (K value: approx. 90) 0.332

Macrogol 35,000 0.635

Water 10,500

M / 45333-PCT Climacteric Polish: 95% carnauba wax

5% bleached wax (e.g., Capol 1295 PH) 0.108

Formulation Example 4: Preparation of Semisolid Dosage Form - Vaginal Gel

The preparation is carried out using conventional methods according to the following two variants:

a) Variant A:

Hydroxypropylmethylcellulose (Hypromellose USP) or another gelling agent is allowed to swell at 2-10% by weight in purified water. Subsequently, the dissolved in glycerol ERR 731 ^® (Preparation Example 1) is incorporated. The amount of glycerol may be up to 50% by weight of the gel. ERr 731 ^® can be dissolved in glycerol up to 0.5% by weight. If necessary, preservatives (eg, sorbic acid and its salts) may be added to the gel. Also a pH adjustment is possible. As an alternative to glycerol, ethanol 30-86% by volume can also be used.

b) Variant B:

Carbomer (Carbopol) is dissolved in purified water at 0.5-5% by weight and the desired pH (eg KOH, NaOH ₁ NH ₃ ) is set. If necessary, a preservative (eg sorbic acid and its salts) is added. After formation of a clear gel ERr 731 ^® (Preparation Example 1) up to 0.5 wt .-% in ethanol from 30 to 86 vol .-% dissolved and added. As an alternative to ethanol, glycerine can also be used.

Formulation Example 5: Preparation of Semisolid Dosage Form - Vaginal Balls

It spheres with a size of 1 to 15 g with a content of 1 to 12 mg ERR 731 ^® (Preparation Example 1) dissolved in glycerol (85% n 20 / D = 1, 45085) prepared by two different variants in a conventional manner.

M / 45333-PCT Climacteric a) Variant A:

Recipe: Gelatin 1 part

Purified water 2 parts

Glycerin 85% (+ ERr ^® 731) 5 parts

b) Variant B: Same recipe, but with a suitable preservative such. Sorbate, benzoate, PHB esters.

The gelatin is each introduced into purified water and allowed to swell until everything has become glassy. Subsequently, glycerol is added 85% with active ingredient and heated, but not above 65 ⁰ C. Then the globules are poured in the usual way

Formulation Example 6: Preparation of Liquid Dosage Form - Drops

a) Lösunqsversuche with ERr 731 ^® in ethanol and glycerol:

Content of the extract:

61, 9% rhaponticin

29.9% deoxyrphaponticin

Experiment A: 200 mg dry extract in 50 ml glycerol R:

55.1% rhaponticin (89.0% of theory)

27.1% deoxyrhaponticin (90.6% of theory)

Experiment B: 200 mg dry extract in 50 ml ethanol 30% R:

52.2% rhaponticin (84.3% of theory) 25.2% deoxyrhaponticin (84.2% of theory)

Experiment C: 200 mg dry extract in 50 ml ethanol 50% R:

58.8% rhaponticin (95.0% of theory)

M / 45333-PCT Climacteric 29.0% deoxyrhaponticin (97.0% of theory)

Experiment D: 200 mg dry extract in 50 ml ethanol 86% R:

59.8% rhaponticin (96.6% of theory)

29.5% deoxyrhaponticin (98.7% of theory)

b) Production of drops:

To prepare drops, dry extract was dissolved in ethanol 30% R according to Experiment B and filtered if necessary.

Test Example 1: In vivo metabolism of the dry extract ERr 731 ^®

To further elucidate the operation of ERr was 731 ^® 3 male and 3 female dogs (kg pure-bred beagles, weight 6-9, age 6-8 months) ERr administered in a dose of 100 mg ^® 731 / kg body weight orally per capsule. At various time intervals, the animals were bled and blood plasma collected. Plasma was assayed for ERr 731 ^® ingredients and metabolites. Surprisingly, significant amounts of the metabolite piceatannol and small amounts of the metabolite resveratrol were detected in both males and females. Maximum plasma levels of these metabolites were reached after approximately 24 h. Plasma levels of piceatannol were significantly higher than those of resveratrol. The test results at 24 h are shown in FIG.

Test Example 2: Study of the activation of the estrogen receptor by ERa ERr 731 ^® and its ingredients or metabolites

The question was whether ERR 731 ^® activates ERa. Therefore, the extract was ERr 731 ^® studied and its ingredients or metabolites (trans-rhapontigenin, Removal soxyrhapontigenin, resveratrol, piceatannol and cis-rhapontigenin) in comparison with 17-beta estradiol on the estrogenic activity in established Modeilsystemen.

Test A):

M / 45333-PCT Climacteric In a first series of experiments, a recombinant yeast screen was used (see E. Routledge and JP Sumpter, Estrogenic activity of surfactants and some of their degradation products assessed using a recombinant yeast screen, Envirom., Tox. Chem.1996).

Saccharomyces cerevisiae cells were stably transfected with human ERα and with a reporter gene consisting of the respective responsive promoter element fused to the LacZ gene which codes for β-galactosidase. By estrogen treatment (with estrogen or an estrogen-like acting substrate) of the cells, mediated by the estrogen receptor, ß-galactosidase is activated, resulting in a red coloration of the yeast cells, which can be measured at 565 nm.

The test results are summarized in FIG.

Test B):

In a second series of experiments, the data of the estrogenicity measurement were verified by determining the induction of alkaline phosphatase in Ishikawa cells (human endometrial adenocarcinoma cells) which had been transfected with an ERa-containing reporter gene construct. The activity of alkaline phosphatase, which is assessed via the chromogenic substrate 4-nitrophenyl phosphate, represents an ERα-mediated response.

The test is based on the description of Holinka CF, Hata H, Kuramoto H, Gurpide E (1986) Effects of Steroid hormones and anti-steroid on alkaline phosphatase activity in human endometrial cancer cells (Ishikawa Line). Cancer Res. 46: 2771-2774, as well as modifications according to Wober J, Weißwange I, Vollmer G (2002) Stimulation of alkaline phosphatase activity in Ishikawa cells induced by various phytoestrogens and synthetic estrogens. J. Steroid Biochem. Mol. Biol. 83: 227-233.

Table 2 presents the concentrations for the positive control (17β-oestradiol) and the test substances used in the assay.

M / 45333-PCT Climacteric Table 2

Exception: The concentration for extract ERr 731 is given in μg / ml

The results of test B) are shown in FIG.

Both tests are valid because all positive and negative controls show their predicted effects (see Figures 2 and 3). The results of both test systems described above show that neither ERr 731 ^® nor its ingredients and metal METABOLITES a significant influence on the activation of ERa comprise (see FIGS. 2 and 3). Thus, it can be assumed that the activity of ERr 731 ^{® is} based on a molecular mechanism independent of ERa.

Test Example 3: Pharmacokinetics of ERr 731 ^® ingredients in female subjects

After oral administration of ERr 731 ^{®, it} should be checked whether one of the components of this combination of active ingredients can be found in the blood to prove that at least one of the components of this combination or its metabolites is bioavailable.

One volunteer took 10 tablets ERr 731 ^® (dosage = 40 mg ERr 731 ^® ) in the morning (8:00 am) with liquid. Then, at various times (as indicated in FIG. 4), 10 ml of blood were taken and the plasma was collected by centrifugation.

M / 45333-PCT Climacteric These plasma samples were worked up as follows: 500 μl of plasma were admixed with 25 μl of an internal standard working solution (2.5 ng / μl rhaponticin or rhapontigenin in methanol) and treated with 500 μl isotonic NaCl solution and 2.5 ml diethyl ether / butanol (9/1; v / v) mixed. After shaking and centrifuging (10 minutes at 4600 rpm), about 2 ml of the supernatant were removed and dried under nitrogen (at 60 ⁰ C). The pellet was taken up in 50 μl of methanol. 200 μl distilled water was added again and 200 μl pipetted into autosampler tubes (light protected). 30 μl of the samples were injected into an LC-MS / MS system (PE Sciex API 3000) for analysis. The chromatographic separation of the analytes was carried out on a Phenomenex Polymer X column with a gradient of an ammonium buffer solution and an acetonitrile / methanol mixture as the mobile phase.

The analysis results are summarized in FIG. Rhaponticin was detected in the blood with a maximum at 3-4 hours (Figure 4), while rhapontigenin could not be found. Since rhaponticin is one of the main ingredients of ERr 731 ^® is assumed that rhaponticin is an activity ingredient of ERr 731 ^®. This is all the more surprising, since it was previously assumed that only the aglycones, but not the glycosylated hydroxystilbenes are effective (Park et al., Arch Pharm Res. 2002; 25: 528-533).

Test Example 4: Estrogen receptor beta (ERß) activation by ERr 731 ^® and its metabolites in the ERß expressing endometrial adenocarcinoma HEC-1B

HEC-1B cells, a human endometrial adenocarcinoma cell line, express neither ERa nor ERβ. Thus, they represent a possibility to investigate ligand-dependent effects of substances on the transactivation mechanisms with respect to different receptor subtypes and different estrogen-responsive promoters in a human and endometrial context.

An established ERβ / mC3 luciferase system was used. These were HEC-1 B

Cells were applied in 24-well plates at a density of 95,000 cells / well (in Duibeck's modified essential medium (DMEM / F12)). The next day these were with

M / 45333-PCT Climacteric an ERß-containing construct (hERβ / pSG2) and a triple-ERE-containing promoter / luciferase construct (mC3-Luc / pGL2) (Hillisch et al., Dissecting physiological roles of estrogen receptor alpha and beta with potent selective ligands from structure- based design. Mol Endocrinol., 2004 Jul; 18: 1599-609). The transient transfection was carried out by means of DOTAP (N- [1- (2,3-dioleoyloxy)] - N, N, N-trimethylammonium propane methyl sulfate; Roth) as described by the manufacturer.

After 24 hours, the cells were treated with appropriate concentrations of the substances or substance mixtures to be investigated. As a positive control served 17ß-estradiol (E2, 10 nM), as a solvent control, a comparable volume of dimethyl sulfoxide (DMSO) was used. The incubation period was 24 hours. Thereafter, the cells were lysed. A hand, by means of the Luciferase Assay ^® kit (Promega), the luciferase activity and on the other hand determined by the BGA ^® kit (Sigma), the protein content. The resulting specific luciferase activities of the substances to be tested were then compared against the DMSO control (100%).

For each test substance at least three transfection experiments are carried out. After calculation of the relative luciferase activity in relation to the negative control (DMSO) of each individual experiment (set as 100%), the corresponding mean values and standard deviations are formed. The presentation of the results is graphically in the form of a bar chart. The significance is calculated using Student's t-test, which was determined as follows: * p <0.05; ** p <0.01; *** p <0.001.

The results for ERr 731 ^® are shown in Figure 5a, for trans-rhapontigenin in Figure 5b and 5c for deoxyrhapontigenin in FIG. They show that the substances activate Dose-dependent ERβ and thus can be used according to the invention.

The experimental results described above demonstrate the surprising finding that, contrary to previous assumptions, the "precursors" of resveratrol and piceatannol, ie the glycosides rhaponticin and deoxyrhaponticin (as main constituents of ERr 731®), can be absorbed per se by human cells has hitherto assumed non-absorbability of these glycosides (compare Park et M / 45333-PCT Climacteric al, Arch. Pharm. Res. 2002, 25 (4), 528-533). Furthermore, the fact that the active ingredient combination ERr 731 ^{® has} higher efficacy than the corresponding aglycones rhapontigenin and deoxyrhapontigenin is surprising. In addition, the latter are not detectable under physiological conditions (experiments not shown).

Thus, by the present invention, the direct pharmacological activity of the glycosides rhaponticin and deoxyrhaponticin (as main components of ERr 731®) could be demonstrated for the first time.

M / 45333-PCT Climacteric

Claims

claims

A solid dosage form comprising a drug-containing solid core with a pharmaceutically acceptable carrier and an active ingredient content of about 1 to 20 wt .-%, based on the total weight of the core, wherein the

Active ingredient comprises a hydroxystilbene-containing active ingredient or a combination of active ingredients containing hydroxystilbene, selected from resveratrol and picacetannol precursors; as well as resveratrol and piceatannol; and their stereoisomeric forms and functional derivatives, in each case in the form of their salts or in the phenol form.

2. Solid dosage form according to claim 1, wherein the active ingredient or the active ingredient combination is obtainable from roots of Rheum rhaponticum.

3. The solid dosage form of claim 1 or 2 wherein the drug combination comprises substantially rhaponticin and deoxyrhaponticin in a weight ratio of about 2: 1 to 1: 2.

4. Solid dosage form according to one of the preceding claims, wherein the active ingredient combination consists essentially of approximately

60-66 wt% Rhaponticin 30-35 wt% Deoxyrhaponticin 0-2 wt% Rhapontigenin; and 0-2% by weight of deoxyrhapontigenin.

5. Solid dosage form according to one of the preceding claims, with a total active substance content of about 2 to 20 mg per dose unit.

A solid dosage form according to any one of the preceding claims, which has a lactose-free core.

A solid dosage form according to any one of the preceding claims in the form of a pill, a tablet, an extrudate or granules.

A solid dosage form according to any one of the preceding claims, which has an enteric coating.

The solid dosage form of claim 8, wherein the coating contains substantially no plasticizer.

M / 45333-PCT Cimacteric

10. A solid dosage form according to any one of the preceding claims having a total weight in the range of about 150 mg ± 20 mg, a core weight of 84 mg ± 10 mg and an active substance content of about 3 to 10 mg per dosage unit.

11. Solid dosage form according to one of the preceding claims, wherein this has a uniformity of the active ingredient content (averaged over 10 or 20 randomly selected individual dosage units) of not more than ± 5 wt .-% based on the total weight of the unit dose.

A process for the preparation of a solid dosage form according to any one of the preceding claims, comprising: a) mixing the active ingredient or combination of active ingredients with the pharmaceutically acceptable carrier; and b) solidifying the mixture to the drug core.

13. The method of claim 12, wherein the active ingredient or combination of active ingredients is dissolved or dispersed in an inert liquid and mixed with the carrier and the solvent removed during or after solidification.

14. The method according to any one of claims 12 and 13, wherein preparing the active ingredient or the active ingredient combination by a) providing a drug-containing part of a drug plant, optionally in comminuted form, b) this is mixed with an aqueous extractant, c ) extracts a liquid extract phase from the mixture after the action of the extractant and, if appropriate, repeatedly repeating the extraction, and d) removing the extractant from the liquid extract phases thus obtained.

15. The method of claim 14, wherein carrying out an extraction with an aqueous extractant at a pH of the mixture in the alkaline range.

16. The method according to claim 14 or 15, wherein the drug plant is selected from plants of the genus Rheum spp, Astragalus spp. Cassia spp or Picea spp.

M / 45333-PCT Cimacteric

17. The method according to any one of claims 12 to 16, wherein the total amount of the active ingredient or the drug combination is added in portions with the pharmaceutically acceptable carrier and mixed after each carrier addition.

18. The method of claim 17, wherein after each addition, mixing is effected with a ball mill over a period of 30 minutes to 3 hours.

19. The method according to any one of claims 12 to 18, wherein providing the drug core with an enteric coating

20. The method of claim 19, wherein the coating contains substantially no plasticizer.

21. The method according to any one of claims 12 to 20, wherein the core is coated.

22. A liquid dosage form comprising an active ingredient or a combination of active substances as defined in any one of claims 1 to 4 in a proportion of about 0.1 to 20 mg / ml in a solvent mixture comprising water and a pharmaceutically acceptable alcohol.

23. The liquid dosage form according to claim 22, wherein the solvent mixture is a water / ethanol mixture having an ethanol content of 10 to 50

Vol .-% is.

24. A liquid dosage form according to any one of claims 22 and 23, formulated as drops for oral administration.

25. Semi-solid dosage form comprising an active ingredient or a combination of active substances as defined in any one of claims 1 to 4 in a proportion of about 1 to 12 wt .-% in a conventional semi-solid carrier.

26. Semi-solid dosage form, formulated as a vaginal gel or vaginal balls.

27. A process for the preparation of a semi-solid or liquid dosage form according to any one of claims 22 to 26, wherein one produces an active ingredient or a combination of active ingredients as defined in claim 14 and formulated in a manner known per se to the dosage form.

M / 45333-PCT Cimacteric

28. A pharmaceutical composition comprising a solid dosage form according to any one of claims 1 to 11 or prepared according to any one of claims 12 to 21 or a liquid or semi-solid dosage form according to any one of claims 22 to 26 or prepared according to claim 27.

29. A solid dosage form according to any one of claims 1 to 11 or prepared according to any one of claims 12 to 21 or a liquid or semi-solid dosage form according to any one of claims 22 to 26 or prepared according to claim 27 for the treatment of climacteric complaints of the woman, juvenile oligomenorrhea and dysmenorrhoea, primary and secondary amenorrhea or endometritis.

30. Use of a solid dosage form according to any one of claims 1 to 11 or prepared according to any one of claims 12 to 21 or a liquid or semi-solid dosage form according to any one of claims 22 to 26 or prepared according to claim 27 for the manufacture of a pharmaceutical composition for the treatment of climacteric complaints of the woman, juvenile oligomenorrhoea and dysmenorrhea, primary and secondary amenorrhoea or endometritis.

31. A dosage form or use according to any one of claims 29 and 30, for the treatment of menopausal symptoms in the peri- or postmenopause.

32. A dosage form or use according to any one of claims 29 and 30, for the treatment of hot flashes, sweats, insomnia, irritability, physical and mental fatigue, sexual problems and urinary tract discomfort.

33. Dosage form or use according to any one of claims 29 and 30, for the treatment of menopausal symptoms due to natural or therapeutically caused menopause.

34. Use of an active substance or a combination of active ingredients according to the above

Definition for selective activation of ERβ in vivo or in vitro.

35. Use of an active substance or a combination of active ingredients according to the above

A definition for treating a disease as defined in any one of claims 30 to 33 by selective activation of ERβ.

M / 45333-PCT Cimacteric