WO2014086480A1

WO2014086480A1 - Lignan compositions

Info

Publication number: WO2014086480A1
Application number: PCT/EP2013/003646
Authority: WO
Inventors: Jianying Yam; Matthias H. Kreuter
Original assignee: Alpinia Laudanum Institute Of Phytopharmaceutical Sciences Ag
Priority date: 2012-12-03
Filing date: 2013-12-03
Publication date: 2014-06-12
Also published as: WO2014086379A1; WO2014086480A8

Abstract

The present invention provides pharmaceutical compositions comprising lignans, which are useful in the treatment or prevention of tumour and/or cancer diseases, in particular for the treatment or prevention of hormone-dependent tumour and/or cancer diseases, such as prostate cancer. Furthermore, the present invention relates to a process for obtaining a lignan composition from lignan-containing plant material, such as from Piper cubeba L., as well as to the use of the lignan composition obtainable by such process as a medicament.

Description

LIGNAN COMPOSITIONS

FIELD OF THE INVENTION

The present invention relates to secondary plant metabolites, in particular to lignan compounds and compositions in the medical field. In particular, the present invention relates to lignan compounds and compositions which are useful in tumour and cancer therapy.

BACKGROUND OF THE INVENTION

Lignan compounds are secondary plant metabolites, which are produced from shikimic acid via the phenylpropanoid pathway. They develop from flavonoid precursors and are responsible for conferring resistance to plants against pathogens and predators. Since lignan compounds are natural products found in various plants, said compounds may be purified from lignan- containing plant material. Furthermore, synthesis processes for the preparation of synthetic lignans are also known for a variety of lignan compounds.

Lignan compounds belong to the group of phytohormones, which have attracted much attention in recent years by the pharmaceutical industry. Phytochemical and biological investigations have been carried out in order to explore the use of lignan compounds in medicine.

The use of medicinal plants for therapeutic purposes is well known throughout many cultures and dates back to the beginning of civilization. A medicinal plant, which is rich in lignans, is Piper cubeba L. of the genus Piperaceae, which is distributed throughout Indonesia in particular the Greater Sunda islands including Borneo, Java and Sumatra. Cultivations of the plant are found in Indonesia, Sri Lanka and India (Assam, Mysore) but also in some African countries such as Sierra Leone and Kongo. To date, twenty four lignans have been reported for Piper cubeba. The berries of Piper cubeba are commonly known as cubeb and used in Indonesian traditional medicine to treat a variety of conditions, such as gonorrhoea, dysentery, syphilis, abdominal pain, diarrhoea, enteritis and asthma.

Cubebs consist of sugars (20 %), proteins (9 %), free amino acids (26.5 %), oils (1_.8.2 %) and several inorganic compounds, above all sulphur (275.9 ppm) and manganese (1 1 6.6 ppm). With regard to secondary metabolites, cubebs contain essential oil compounds, various lignans and fatty oil with palmitinic, oleic, linolic and linolenic acid being the major components. Furthermore, cubebic acid as well as acidic and neutral resins (3.0 to 3.7 %) were found in cubebs.

Only three groups of secondary metabolites have been reported from the berries of Piper cubeba, i.e. alkaloids, lignans and terpenoids (essential oil). The lignans and the essential oil have been more intensively investigated, since Piper cubeba accumulates both groups of compounds in relatively high amounts.

Elfami et al. showed that the leaves and stalks of P. cubeba contain more than 20 different lignans, with yatein, hinokinin and cubebin as the major lignans in the berries, leaves and stalks of P. cubeba (Elfami et al., Biochemical Systematics and Ecology 35 (2007) 397-402).

Of the lignan compounds found in Piper cubeba, yatein, hinokinin, cubebin and dihydrocubebin have been reported to have antifeedant activity against a number of stored product insects. This activity is comparable to that of podophyllotoxin. Moreover, several lignan compounds have been reported to possess therapeutic activity. Thus, e.g. hinokinin has been reported to have an anti-inflammatory and analgesic effect. Lignans have also been reported to inhibit the activity of cytochrome P450-class enzymes that are involved in the metabolism of a wide range currently used drugs and chemotherapeutics.

Cubebin and cubebin derivatives have been described to be useful in Chagas' disease prophylaxis (cf. WO 03/080600 A1 ) and for the treatment and prevention of liver disease (cf. JP 1 180824A). Furthermore, analgesic and anti-inflammatory activities and their usefulness for the treatment of pain and inflammatory diseases, such as arthritis, have been suggested for cubebin derivatives and hinokinin (cf. WO 2006/1 13981 A2). Pusztai et al. (Pusztai et al., 2010, Anticancer Research 30: 451 -454) investigated a range of lignans as potential cancer chemopreventive agents by evaluating their ability to inhibit HCMV immediate early (IE) antigen expression in A549 cells. While hinokinin was able to reduce IE- antigen expression, dihydrocubebin even resulted in an increase in IE-antigen expression, which renders dihydrocubebin unsuitable as chemopreventive agent in this model system.

Mansoor et al. (Mansoor et al., 2012, Phytotherapy Res. 26: 292-696) evaluated seven lignans obtained from Pycnanthus angolensis or by derivatisation for their ability to induce apoptosis in human HuH-7 hepatoma cells. The results indicated that among others hinokinin appeared to be an inducer of apoptosis in HuH-7 cells. Furthermore, hinokinin appeared to possess cytotoxic activity against the P-388 and HT-29 cell lines in the study of Lin et al. (cf. Lin et al., 2004, Planta Med 70: 234-238), however, according to a study of Chang et al. (Chang et al., 2000, Phytochemistry 55: 227-232) who consider compounds with ED₅₀ values of less than 4 pg/ml active in this respect, hinokinin does not show any significant cytotoxic effects on the A- 549, MCF-7 and HT-29 cell lines.

The antiproliferative activities of yatein and yatein derivatives on the P-388, A549, MCF-7 and HT-29 cell lines was assessed by Medarde et al. (Medarde et al., 1995, Arch. Pharm. (Weinheim) 328: 403-407 and 640-644). Furthermore, Novelo et al. (Novelo et al., 1993, Journal of Natural Plant Products 56: 1 728-1 736) evaluated the cytotoxic potential of several lignans, among them yatein, with a number of human cell lines, in particular with the A431 , BC1 , Col-2, HT, KB, KB-V1 , LNCaP, Lu-1 , U373 and ZR-75-1 cell lines. According to Novelo et al., a group of lignan compounds, including yatein, demonstrated a non-specific cytotoxic activity.

The usefulness of medicinal plant extracts for inhibition of tumour cell growth has also been assessed. For example, WO 2009/021347 A1 and the accompanying publication of Yam et al. (Yam et al., 2008, Planta Med.: 74: 33-38) describe the use of extracts from Piper cubeba L. as an effective component in a drug for the treatment of cancer diseases. It was also shown that cubebin, a component of the Piper cubeba extract, possesses some anti-proliferative activity on the LNCaP and PC-3 cell lines.

Although several studies on the therapeutic value of lignans have been performed to date, therapeutically useful and effective lignan compositions for use in medicine are still sparsely found in the pharmaceutical sector. Thus, there is a need for novel pharmaceutical compositions based on secondary plant metabolites, such as lignans, which are therapeutically effective and, thus, useful in medicine.

Accordingly, it is an object of the present invention to provide therapeutically effective compositions based on secondary plant metabolites, in particular based on lignan compounds. Such compositions are preferably useful in the prevention or treatment of tumour diseases, preferably in the prevention or treatment of hormone-dependent tumour diseases, e.g. in the prevention or treatment of prostate cancer. Furthermore, it is an object of the present invention to provide pharmaceutical compositions based on secondary plant metabolites, in particular based on lignan compounds, which exhibit good bioavailability, in particular good oral bioavailability.

The objects of the present invention are solved by the subject-matter of the appended claims.

SUMMARY OF THE INVENTION

The present inventors were able to show that a particular subset of lignan compounds, e.g. present in Piper cubeba, i.e. clusin, dihydrocubebin, hinokinin, yatein and cubebin, exhibit, each individually, a surprisingly effective therapeutic activity, in particular in the prevention and treatment of tumour diseases, such as prostate cancer, breast cancer and pancreatic cancer, which is even superior if the lignan compounds are used in a combination of at least two of the lignans of the identified subset of lignan compounds. The inventors also surprisingly found that the therapeutic activity of the inventive lignan compounds on prostate cancer types is even further enhanced when said lignan compounds are used in a combination of at least three, e.g. three, four, or five of the lignans of the identified subset of lignan compounds.

Furthermore, the present inventors found that the combined application of at least two of the lignan compounds of the identified subset of lignan compounds results in unexpectedly improved bioavailability of the single lignan compounds, in particular in an improved oral bioavailability.

Accordingly, the above objects of the present invention are solved by a pharmaceutical composition comprising at least one of the compounds, preferably at least two, more preferred at least three of the compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, preferably for use as a medicament. Accordingly, pharmaceutical compositions according to the present invention comprise yatein and clusin and at least one further compound selected from the group consisting of dihydrocubebin, hinokinin and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, preferably for use as a medicament.

Furthermore, the present invention provides pharmaceutical compositions comprising at least three compounds selected from the group consisting of clusin, dihydrocubebin, yatein and cubebin, optionally comprising one or two further lignans from said group, such that the composition according to the invention comprises at least three, optionally 4 or all of the lignans from said group and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, preferably for use as a medicament.

The present invention furthermore provides pharmaceutical compositions comprising clusin, yatein and dihydrocubebin and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, preferably for use as a medicament.

Further, the present invention provides pharmaceutical compositions comprising at least four compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, preferably for use as a medicament.

The present invention also provides pharmaceutical compositions comprising at least two, but not more than four compounds selected from the group comprising clusin, dihydrocubebin, hinokinin, yatein and cubebin, and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, preferably for use as a medicament.

While the compositions according to the present invention are useful for the treatment or prevention of a variety of conditions and diseases, the pharmaceutical compositions according to the present invention are particularly useful in the treatment of tumour diseases, such as tumour diseases and cancers, which are influenced by hormones in their growth and phenotype, such as, e.g. hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone- dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer. Thus, in a preferred embodiment, the pharmaceutical compositions according to the present invention are for use in the treatment or prevention of tumour diseases, preferably for use in the treatment or prevention of cancer diseases, more preferably for use in the treatment or prevention of hormone-dependent cancer diseases, such as in the treatment or prevention of prostate cancer.

Accordingly, the present invention provides clusin, yatein and/or dihydrocubein for use in the treatment or prevention of a tumour disease, such as, e.g. prostate cancer. Clusin and yatein and/or dihydrocubein were shown to exhibit a particularly effective therapeutic activity. Accordingly, the present invention provides clusin, yatein and/or dihydrocubein for use as a medicament, in particular for use in the treatment or prevention of tumour diseases, preferably in the treatment or prevention of cancer diseases, more preferably for use in the treatment or prevention of hormone-dependent cancer diseases, such as in the treatment or prevention of prostate cancer.

Furthermore, the present invention provides clusin, dihydrocubebin and/or hinokinin for use in the treatment or prevention of a tumour disease, such as prostate cancer. Dihydrocubebin and clusin were shown by the present inventors to exhibit a particularly effective therapeutic activity. Accordingly, the present invention provides dihydrocubebin and/or clusin for use as a medicament, in particular for use in the treatment or prevention of tumour diseases, preferably in the treatment or prevention of cancer diseases, more preferably for use in the treatment or prevention of hormone-dependent cancer diseases, such as in the treatment or prevention of prostate cancer.

The present invention further provides clusin, dihydrocubein and/or cubebin for use in the treatment or prevention of a tumour disease, such as prostate cancer. Clusin, dihydrocubein and/or cubebin were shown to exhibit a particularly effective therapeutic activity. Accordingly, the present invention provides clusin, dihydrocubein and/or cubebin for use as a medicament, in particular for use in the treatment or prevention of tumour diseases, preferably in the treatment or prevention of cancer diseases, more preferably for use in the treatment or prevention of hormone-dependent cancer diseases, such as in the treatment or prevention of prostate cancer.

The present invention further provides clusin, dihydrocubebin, yatein, and/or hinokinin for use in the treatment or prevention of a tumour disease, such as prostate cancer. Clusin, yatein, dihydrocubebin and/or hinokinin were shown to exhibit a particularly effective therapeutic activity. Accordingly, the present invention provides clusin, yatein, dihydrocubebin or hinokinin, or clusin, yatein, dihydrocubebin and hinokinin for use as a medicament, in particular for use in the treatment or prevention of tumour diseases, preferably in the treatment or prevention of cancer diseases, more preferably for use in the treatment or prevention of hormone^dependent cancer diseases, such as in the treatment or prevention of prostate cancer.

Furthermore, the present invention provides dihydrocubebin, yatein and/or hinokinin for use in the treatment or prevention of a tumour disease, such as prostate cancer. Dihydrocubebin, yatein and/or hinokinin were shown to exhibit a particularly effective therapeutic activity. Accordingly, the present invention provides clusin, yatein, dihydrocubebin and/or hinokinin for use as a medicament, in particular for use in the treatment or prevention of tumour diseases, preferably in the treatment or prevention of cancer diseases, more preferably for use in the treatment or prevention of hormone-dependent cancer diseases, such as in the treatment or prevention of prostate cancer.

The present invention further provides clusin, dihydrocubebin, yatein and hinokinin for use in the treatment or prevention of a tumour disease, such as prostate cancer. Clusin, dihydrocubebin, yatein and hinokinin were shown to exhibit a particularly effective therapeutic activity. Accordingly, the present invention provides clusin, dihydrocubebin, yatein and hinokinin for use as a medicament, in particular for use in the treatment or prevention of tumour diseases, preferably in the treatment or prevention of cancer diseases, more preferably for use in the treatment or prevention of hormone-dependent cancer diseases, such as in the treatment or prevention of prostate cancer.

The present invention further provides clusin, dihydrocubebin, yatein, hinokinin and cubebin for use in the treatment or prevention of a tumour disease, such as prostate cancer. Clusin, dihydrocubebin, yatein and hinokinin were shown to exhibit a particularly effective therapeutic activity. Accordingly, the present invention provides . Clusin, dihydrocubebin, yatein and hinokinin for use as a medicament, in particular for use in the treatment or prevention of tumour diseases, preferably in the treatment or prevention of cancer diseases, more preferably for use in the treatment or prevention of hormone-dependent cancer diseases, such as in the treatment or prevention of prostate cancer.

Furthermore, the present invention provides compositions comprising at least three, preferably at least four, most preferred at least five lignans selected from the group comprising clusin, yatein, dihydrocubebin, hinokinin and cubebin for use in aiding in the prophylaxis or prevention of cancer, more preferably for use in aiding in the prophylaxis or prevention of hormone-dependent cancer diseases, such as in aiding in the prophylaxsis or prevention of prostate cancer. The present invention also provides a process for preparing lignan compositions, in which the concentration of lignans is at least 65% (w/w), more preferably at least 70% (w/w) from lignan- containing plant material.

BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1 : Anti-proliferative effects of cubebin, clusin, dihydrocubebin, hinokinin and yatein were assessed on the human prostate cancer cell line LNCaP. A solvent control was used at 1 % (v/v) ethanol final concentration.

Fig. 2: Anti-proliferative effects of 10pg/ml of the lignan fraction (Pip.cub.40p2.001 .E03.S04; cf. Example 1 1 ) and the five individual lignans on cells of the LNCaP cell line were assessed. A solvent control was used at 1 % (v/v) ethanol final concentration. The dose response curves and the IC₅₀ values of the lignans are presented in the graph.

Fig. 3: Anti-proliferative effects of lignan-spiked compositions according to Example 20 were assessed on cells of the human prostate cancer cell line LNCaP. 1 % (v/v) ethanol final concentration was used as solvent control; 3A: cubebin and dihydrocubebin, 3B: clusin and hinokinin, 3C: yatein.

Fig. 4: Effect of 10pg/ml of the indicated lignan compositions on LNCaP cell growth. The individual lignans and the lignan fraction (Pip.cub.40p2.001 .E03.S04; cf. Example 1 1 ) were tested alone and in combination with various lignans (lignan-spiked fractions) as indicated. All data points are expressed as % of the solvent controls.

Fig. 5: Pharmacokinetics of hinokinin and lignan fraction pip.cub.40p2.001 .E03.S04 (Example 1 1 ) in the plasma of male SCID-beige mice. Fig. 6: Tissue distribution of lignans in murine organs of male SCID-beige mice analyzed by HPLC following oral administration.

Fig. 7: Anti-proliferative effects of 10pg/ml of the lignan fraction (Pip.cub.40p2.001 .E03.S04; cf. Example 1 1 ) and the five individual lignans on cells of the HPAFII cell line were assessed. A solvent control was used at 1 % (v/v) ethanol final concentration. The dose response curves and the IC₅₀ values of the lignans are presented in the graph. Fig. 8: Anti-proliferative effects of different lignan combinations on the prostate cancer cell lines LNCaP, PC-3M cell. LNCap and PC-3 cells represent hormone-dependent (LNCaP), i.e. androgen-sensitive and hormone-insensitive (PC-3M) cell cancer types, respectively. Lignan combinations and lignan fraction Pip.cub.40p2.001 .E03.S04 (cf. Example 1 1 ) were applied as indicated to (A) LNCaP and (B) PC-3M cells at a final concentration of 10pg/ml total lignans, whereby the individual lignan concentration was 5pg/ml, 3,3pg/ml and 2,5pg/ml if two, three or four lignans were used in the lignan combination. DETAILED DESCRIPTION OF THE INVENTION

Although the present invention is described in detail below, it is to be understood that this invention is not limited to the particular methodologies, protocols and reagents described herein as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims. Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art.

In the following, the elements of the present invention will be described. These elements are listed with specific embodiments, however, it should be understood that they may be combined in any manner and in any number to create additional embodiments. The variously described examples and preferred embodiments should not be construed to limit the present invention to only the explicitly described embodiments. This description should be understood to support and encompass embodiments which combine the explicitly described embodiments with any number of the disclosed and/or preferred elements. Furthermore, any permutations and combinations of all described elements in this application should be considered disclosed by the description of the present application unless the context indicates otherwise.

Throughout this specification and the claims which follow, unless the context requires otherwise, the term "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated member, integer or step and the optional inclusion of any other non-stated member, integer or step. The term "consist of" is a particular embodiment of the term "comprise", wherein any other non-stated member, integer or step is excluded. In the context of the present invention, the term "comprise" encompasses the term "consist of". The terms "a" and "an" and "the" and similar references used in the context of describing the invention (especially in the context of the claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention. Several documents are cited throughout the text of this specification. Each of the documents cited herein (including all patents, patent applications, scientific publications, manufacturer's specifications, instructions, etc.), whether supra or infra, are hereby incorporated by reference in their entirety. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

The present invention is based on the surprising finding that the lignan compounds of a particular subset of lignan compounds, e.g. obtainable from lignan-containing plant material, such as from the fruits of Piper cubeba, i.e. the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin, provide individually excellent therapeutic potential, in particular for the treatment or prevention of tumour diseases, and that the combination of two or more, preferably three or moreof said lignan compounds provides even superior therapeutic efficiency, e.g. in tumour therapy unexpectedly high bioavailability of individual lignan compounds when administered in combination, in particular excellent oral bioavailability The lignan compounds of the subset of lignan compounds according to the present invention preferably exhibit chemical structures according to Formulas I to V below:

(V) Yatein

In the context of the present invention, any stereochemical variant or a mixture of stereochemical variants of the lignan compounds used in the present invention are encompassed. For example, lignan compound "clusin" includes (-)-clusin, (+)-clusin, and mixtures of (-)- and (+)-clusin, lignan compound "dihydrocubebin" includes (-)-dihydrocubebin, (+)-dihydrocubebin, and mixtures of (-)- and (+)-dihydrocubebin, lignan compound "hinokinin" includes (-)-hinokinin, (+)-hinokinin, and mixtures of (-)- and (+)-hinokinin, lignan compound "yatein" includes (-)-yatein, (+)-yatein, and mixtures of (-)- and (+)-yatein, lignan compound "cubebin" includes (-)-cubebin, (+)-cubebin, and mixtures of (-)- and (+)-cubebin. In a preferred embodiment, the lignan compound is the (-)-lignan, i.e. (-)-clusin, (-)-dihydrocubebin, (-)- hinokinin, (-)-yatein, (-)-cubebin.

The lignan compounds used in the present invention may be obtained by isolation and purification from natural sources or by de novo synthesis or semi synthesis from suitable starting materials. For example, the lignan compounds may be obtained by purification from plant extracts, e.g. extracts of defatted powder of Piper cubeba L. or Piper clusii Cass., by filtration and direct application of ethanolic extracts on preparative reversed phase chromatography employing RP-C18 silica and elution under isocratic conditions using 70% methanol aq. as eluent and subsequent further purification by high pressure liquid chromatography (HPLC). A detailed description of methods for obtaining clusin, dihydrocubebin, cubebin, hinokinin and yatein from lignan-containing plant material is provided in the examples of the present disclosure.

Alternatively, the lignan compounds used in the present invention are obtainable by synthesis. For example, hinokinin or dihydrocubebin are obtainable by oxidative homocoupling of chiral 3-arylpropanoic acid derivates as, e.g. described by Kise et al. (Kise et al., 2000, J. Org. Chem. 65: 464-468). The synthesis of cubebin and yatein is also described in the literature (Batterbee et al., 1969, J. Chem. Soc. (C): 2470-2477; Neidigh et al., 1 994, Journal of Natural Products 57: 791 - 800). A detailed report on the asymmetric synthesis of dibenzylbutane- and dibenzylbutyrolactone-type lignans has been published by Enders and Milovanovic (Enders and Milovanovic, 2007, Z. Naturforsch. 62b: 1 1 7-120; Ph.D. thesis: Mile Milovanovic, Rheinisch- Westfalischen Technischen Hochschule, Aachen, Germany 2007 „Asymmetrische Synthese von Dibenzylbutan- und Dibenzylbutyrolacton-Lignanen", ISBN 3-86130-559-3). Thus, the feasibility of synthesis and, thus, accessibility of the individual compounds used in the present invention has also been demonstrated in the above cited publications. The lignan compounds used in the present invention are individually suitable for use in medicine, in particular for use in the treatment or prevention of tumour diseases. Accordingly, preferably the lignan compounds used in the present invention as well as the pharmaceutical compositions according to the present invention and the lignan compositions according to the present invention are preferably for use in tumour therapy, i.e. for use in the treatment or prevention of tumour diseases, wherein "treatment" refers to a therapy of an existing tumour disease and "prevention" refers to preventive therapy for prevention of the occurrence of a tumour disease.

According to the invention, the term "tumour" or "tumour disease" refers to a swelling or lesion formed by an abnormal growth of cells (called neoplastic cells or tumour cells). By "tumour cell" is meant an abnormal cell that grows by a rapid, uncontrolled cellular proliferation and continues to grow after the stimuli that initiated the new growth cease. Tumours show partial or complete lack of structural organization and functional coordination with the normal tissue, and usually form a distinct mass of tissue, which may be benign, pre-malignant, or malignant.

Preferably, a tumour disease in the context of the invention is a cancer disease, i.e. a malignant disease, and a tumour cell is a cancer cell. Preferably, a tumour disease according to the invention is cancer, wherein the term "cancer" according to the invention preferably comprises leukaemias, seminomas, melanomas, teratomas, lymphomas, neuroblastomas, glioblastomas, rectal cancer, endometrial cancer, kidney cancer, adrenal cancer, thyroid cancer, blood cancer, skin cancer, cancer of the brain, cervical cancer, intestinal cancer, liver cancer, colon cancer, stomach cancer, head and neck cancer, gastrointestinal cancer, lymph node cancer, esophagus cancer, colorectal cancer, pancreatic cancer, ear, nose and throat (ENT) cancer, breast cancer, prostate cancer, cancer of the uterus, ovarian cancer and lung cancer and the metastases thereof. Examples thereof are breast cancer, such as mamma carcinomas, prostate cancer, such as prostate carcinomas, and pancreatic cancers or metastases of the cancer types or tumours described above. Preferably, the tumour disease in the context of the present invention is selected from the group consisting of prostate cancer, breast cancer, and pancreatic cancer, most preferably the tumour disease in the context of the present invention is prostate cancer, such as hormone-dependent prostate cancer.

The "tumour disease" and "cancer" in the context of the present invention may be hormone- dependent or hormone-independent. Preferably, "tumour disease" and "cancer" in the context of the present invention is hormone-dependent. The term "hormone-dependent" tumour diseases and cancers relates to tumour diseases and cancer types, which are influenced by hormones in their growth and phenotype, such as, e.g. hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone- dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer. Thus, said term refers to any tumour disease or cancer, in which cellular functions, such as cell division of the respective cancer or tumour cells, are hormone-dependent.

The term "hormone" in the context of the invention preferably refers to sex hormones, such as e.g. androgens and estrogens, but also includes steroid hormones and peptide hormones. Thus, preferably, the term "hormone-dependent" refers to androgen- or estrogen-dependency. A particularly preferred hormone-dependent cancer in the context of the present invention is hormone-dependent prostate cancer.

The term "androgen" in the context of the invention refers, for example, to dehydroepiandrosterone, androstenedione, androstenediol, dihydrotestosterone and teststosterone. The term "estrogen" in the context of the invention refers, for example, to estrone, estradiol and estriol.

The term "hormone-independent" tumour disease or cancer in the context of the present invention refers to any tumour disease or cancer, which is not hormone-dependent for its growth or respective phenotype, such as e.g. retinoblastoma etc..

The lignan compounds used in the present invention are particularly superior in their therapeutic activity, e.g. for treating or preventing tumour diseases, if at least two of the lignan compounds according to the present invention, i.e. two, three, four or five lignan compounds according to the present invention are used in combination. Furthermore, it was surprisingly found that the bioavailability of the single lignans is improved if at least two lignans of the subset of lignans used in the present invention, i.e. clusin, dihydrocubebin, hinokinin, yatein and cubebin, are used in combination.

Thus, for example, combinations of clusin and dihydrocubebin, clusin and hinokinin, clusin and yatein, clusin and cubebin, dihydrocubebin and hinokinin, dihydrocubebin and yatein, dihydrocubebin and cubebin, hinokinin and yatein, hinokinin and cubebin, and yatein and cubebin are combinations of two lignan compounds according to the present invention, combinations of clusin, dihydrocubebin and hinokinin, clusin, dihydrocubebin and yatein, clusin, dihydrocubebin and cubebin, clusin, hinokinin and yatein, clusin, hinokinin and cubebin, clusin, yatein and cubebin, dihydrocubebin, hinokinin and yatein, dihydrocubebin, hinokinin and cubebin, dihydrocubebin, yatein and cubebin, and hinokinin, yatein and cubebin are combinations of three lignan compounds according to the present invention, combinations of clusin, dihydrocubebin, hinokinin and yatein, clusin dihydrocubebin, hinokinin and cubebin, clusin, hinokinin, yatein and cubebin, clusin, dihydrocubebin, yatein and cubebin, and dihydrocubebin, hinokinin, yatein and cubebin are combinations of four lignan compounds according to the present invention. Particularly preferred combinations according to the present invention comprise clusin and/or dihydrocubebin, or clusin and/or yatein. Particularly preferred combinations of three lignans according to the present invention comprise clusin, dihydrocubebin and yatein, or clusin, dihydrocubebin and cubebin. Particularly preferred combinations of four lignans lignans according to the present invention comprise clusin, dihydrocubebin, yatein and hinokinin.

In one aspect, the present invention provides a pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents. The at least one compound is preferably selected from the group consisting of clusin, dihydrocubebin and hinokinin, more preferably from the group consisting of clusin and dihydrocubebin, more preferably clusin and yatein.

The pharmaceutical composition according to the present invention is preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein. The tumour disease is preferably cancer, which may be hormone-dependent or hormone-independent. Preferably, the cancer or tumour disease is hormone-dependent, such as estrogen- or androgen-dependent. For example, the cancer or tumour disease is selected from the group consisting of hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer.

In preferred embodiments, the pharmaceutical composition according to the present invention is for use in the treatment or prevention of prostate cancer, breast cancer or pancreatic cancer, preferably for use in the treatment of prostate cancer, such as hormone-dependent prostate cancer. Thus, for example, the present invention provides a pharmaceutical composition comprising clusin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising clusin according to the present invention further comprises at least one compound selected from the group consisting of dihydrocubebin, hinokinin, yatein and cubebin. In one embodiment, the pharmaceutical composition according to the present invention comprises clusin as the only pharmaceutically active ingredient. In one embodiment, the pharmaceutical composition consists of clusin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Clusin is preferably present in the pharmaceutical composition according to the present invention in a concentration of at least 10% (w/w), preferably at least 20% (w/w), more preferably at least 30% (w/w), such as at least 40% (w/w), at least 50% (w/w), or at least 60% (w/w), and may be present in a concentration of up to 100%, such as in a concentration of 90% (w/w), 95% (w/w), 99% (w/w) or 100%. Thus, clusin is preferably present in the pharmaceutical composition according to the present invention in a concentration of between about 10% and 100% (w/w), such as in a concentration of between about 10% and 99% (w/w), 10% and 95% (w/w), 10% and 90% (w/w), 10% and 60% (w/w), or 10% and 30% (w/w).

The present invention further provides a pharmaceutical composition comprising dihydrocubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising dihydrocubebin according to the present invention further comprises at least one compound selected from the group consisting of clusin, hinokinin, yatein and cubebin. In one embodiment, the pharmaceutical composition comprises dihydrocubebin as the only pharmaceutically active ingredient. In one embodiment, the pharmaceutical composition according the present invention consists of dihydrocubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Dihydrocubebin is preferably present in the pharmaceutical composition according to the present invention in a concentration of at least 5% (w/w), preferably at least 10% (w/w), more preferably at least 15% (w/w), such as at least 20% (w/w), at least 25% (w/w), or at least 30% (w/w), and may be present in a concentration of up to 100%, such as in a concentration of 90% (w/w), 95% (w/w), 99% (w/w) or 100%. Thus, dihydrocubebin is preferably present in the pharmaceutical composition according to the present invention in a concentration of between about 5% and 100% (w/w), such as in a concentration of between about 5% and 99% (w/w), 5% and 95% (w/w), 5% and 90% (w/w), 5% and 50% (w/w), or 5% and 25% (w/w).

The present invention further provides a pharmaceutical composition comprising hinokinin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising hinokinin according to the present invention further comprises at least one compound selected from the group consisting of clusin, dihydrocubebin, yatein and cubebin. In one embodiment, the pharmaceutical composition comprises hinokinin as the only pharmaceutically active ingredient. In one embodiment, the pharmaceutical composition according to the present invention consists of hinokinin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Hinokinin is preferably present in the pharmaceutical composition according to the present invention in a concentration of at least 1 5% (w/w), preferably at least 20% (w/w), more preferably at least 30% (w/w), such as at least 40% (w/w), at least 50% (w/w), or at least 60% (w/w), and may be present in a concentration of up to 100%, such as in a concentration of 90% (w/w), 95% (w/w), 99% (w/w) or 100%. Thus, hinokinin is preferably present in the pharmaceutical composition according to the present invention in a concentration of between about 1 5% and 100% (w/w), such as in a concentration of between about 15% and 99% (w/w), 15% and 95% (w/w), 15% and 90% (w/w), 15% and 70% (w/w), or 15% and 50% (w/w).

The present invention further provides a pharmaceutical composition comprising yatein and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising yatein according to the present invention further comprises at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin and cubebin. In one embodiment, the pharmaceutical composition comprises yatein as the only pharmaceutically active ingredient. In one embodiment, the pharmaceutical composition according the present invention consists of yatein and one or more pharmaceutically acceptable excipients, carriers, and/or diluents.

Yatein is preferably present in the pharmaceutical composition according to the present invention in a concentration of at least 2% (w/w), preferably at least 4% (w/w), more preferably at least 8% (w/w), such as at least 1 5% (w/w), at least 30% (w/w), or at least 50% (w/w), and may be present in a concentration of up to 100%, such as in a concentration of 90% (w/w), 95% (w/w), 99% (w/w) or 100%. Thus, yatein is preferably present in the pharmaceutical composition according to the present invention in a concentration of between 2% and 100% (w/w), such as in a concentration of between about 2% and 99% (w/w), 2% and 95% (w/w), 2% and 90% (w/w), 2% and 50% (w/w), or 2% and 15% (w/w).

The present invention further provides a pharmaceutical composition comprising cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising cubebin according to the present invention further comprises at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin and yatein. In one embodiment, the pharmaceutical composition comprises cubebin as the only pharmaceutically active ingredient. In one embodiment, the pharmaceutical composition according the present invention consists of cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Cubebin is preferably present in the pharmaceutical composition according to the present invention in a concentration of at least 25% (w/w), preferably at least 35% (w/w), more preferably at least 45% (w/w), such as at least 55% (w/w), at least 65% (w/w), or at least 75% (w/w), and may be present in a concentration of up to 100%, such as in a concentration of 90% (w/w), 95% (w/w), 99% (w/w) or 100%. Thus, cubebin is preferably present in the pharmaceutical composition according to the present invention in a concentration of between 25% and 100% (w/w), such as in a concentration of between about 25% and 99% (w/w), 25% and 95% (w/w), 25% and 90% (w/w), 25% and 80% (w/w), or 25% and 60% (w/w).

Particularly preferred pharmaceutical compositions according to the present invention are those comprising clusin and/or dihydrocubebin, preferably for use as a medicament, more preferably for use in the treatment of a tumour disease as disclosed herein.

In a preferred embodiment, the pharmaceutical composition according to the present invention comprises at least two compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, preferably from the group consisting of clusin, dihydrocubebin and hinokinin.

Furthermore, in a preferred embodiment, the pharmaceutical composition according to the present invention comprises at least two compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein, preferably from the group consisting of clusin, dihydrocubebin and yatein.

Preferred pharmaceutical compositions according to the present invention are those comprising clusin, hinokinin and/or yatein, preferably for use as a medicament, more preferably for use in the treatment of a tumour disease as disclosed herein.

Particularly preferred pharmaceutical compositions according to the present invention are those comprising three lignans selected from the group consisiting of clusin, dihydrocubebin, yatein and cubebin, preferably for use as a medicament, more preferably for use in the treatment of a tumour disease as disclosed herein. Accordingly, the particularly preferred pharmaceutical compositions of the present invention may comprise clusin, dihydrocubebin and yatein, or dihydrocubebin, yatein and cubebin, or clusin, yatein and cubebin, or clusin, dihydrocubebin and cubebin, preferably for use as a medikament, , more preferably for use in the treatment of a tumour disease as disclosed herein.

Particularly preferred pharmaceutical compositions according to the present invention are those comprising yatein and clusin and one further compound selected from the group consisiting of dihydrocubebin, hinokinin and cubebin, preferably for use as a medicament, more preferably for use in the treatment of a tumour disease as disclosed herein. Accordingly, particularly preferred pharmaceutical compositions according to the present invention comprise yatein, clusin and dihydrocubebin, or yatein, clusin and hinokinin, or yatein, clusin and cubebin, preferably for use as a medikament, , more preferably for use in the treatment of a tumour disease as disclosed herein.

Particularly preferred pharmaceutical compositions according to the present invention are those comprising dihydrocubebin and hinokinin and/or yatein, preferably for use as a medicament, more preferably for use in the treatment of a tumour disease as disclosed herein.

Particularly preferred pharmaceutical compositions according to the present invention are those comprising clusin, dihydrocubebin and yatein, preferably for use as a medicament, more preferably for use in the treatment of a tumour disease as disclosed herein

Particularly preferred pharmaceutical compositions according to the present invention are those comprising clusin, dihydrocubebin, hinokinin and/or yatein, preferably for use as a medicament, more preferably for use in the treatment of a tumour disease as disclosed herein. Particularly preferred pharmaceutical compositions according to the present invention are those comprising clusin, dihydrocubebin, yatein and cubebin for use as a medicament; more preferably for use in the treatment of a tumour disease, more preferably for use in the treatment of a hormone-independent tumour as disclosed herein. Any combination of the lignan compounds as disclosed herein may be comprised in the pharmaceutical composition according to the present invention or in a composition for use in aiding in the prophylaxsis or prevention of cancer according to the present invention.

Thus, the present invention provides, for example, a pharmaceutical composition comprising clusin and dihydrocubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising clusin and dihydrocubebin further comprises a compound selected from the group consisting of hinokinin, yatein and cubebin. In one embodiment, the pharmaceutical composition comprises clusin and dihydrocubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, dihydrocubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. One or .both of clusin and dihydrocubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or dihydrocubebin may be present in a concentration of, e.g. at least 5% (w/w), such as in a concentration of between 5% and 20% (w/w), in the pharmaceutical composition according to the present invention. Furthermore, the present invention provides a pharmaceutical composition comprising clusin and hinokinin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising clusin and hinokinin further comprises a compound selected from the group consisting of dihydrocubebin, yatein and cubebin. In one embodiment, the pharmaceutical composition comprises clusin and hinokinin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, hinokinin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. One or both of clusin and hinokinin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or hinokinin may be present in a concentration of, e.g. at least 1 5% (w/w), such as in a concentration of between 15% and 50% (w/w), in the pharmaceutical composition according to the present invention. Furthermore, the present invention provides a pharmaceutical composition comprising clusin and yatein and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising clusin and yatein further comprises a compound selected from the group consisting of dihydrocubebin, hinokinin and cubebin. In one embodiment, the pharmaceutical composition comprises clusin and yatein as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, yatein and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. One or both of clusin and yatein may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2% (w/w), such as in a concentration of between 2% and 1 5% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising clusin and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising clusin and cubebin further comprises a compound selected from the group consisting of dihydrocubebin, hinokinin and yatein. In one embodiment, the pharmaceutical composition comprises clusin and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. One or both of clusin and cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25% (w/w), such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising dihydrocubebin and hinokinin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising dihydrocubebin and hinokinin further comprises a compound selected from the group consisting of clusin, yatein and cubebin. In one embodiment, the pharmaceutical composition comprises dihydrocubebin and hinokinin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of dihydrocubebin, hinokinin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. One or both of dihydrocubebin and hinokinin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, dihydrocubebin may be present in a concentration of at least 5% (w/w), e.g. in a concentration of between 5% and 20% (w/w), and/or hinokinin may be present in a concentration of, e.g. at least 15% (w/w), such as in a concentration of between 15% and 50% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising dihydrocubebin and yatein and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising dihydrocubebin and yatein further comprises a compound selected from the group consisting of clusin, hinokinin and cubebin. In one embodiment, the pharmaceutical composition comprises dihydrocubebin and yatein as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of dihydrocubebin, yatein and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. One or both of dihydrocubebin and yatein may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, dihydrocubebin may be present in a concentration of at least 5% (w/w), e.g. in a concentration of between 5% and 20% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2% (w/w), such as in a concentration of between 2% and 15% (w/w), in the pharmaceutical composition according to the present invention. Furthermore, the present invention provides a pharmaceutical composition comprising dihydrocubebin and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising dihydrocubebin and cubebin further comprises a compound selected from the group consisting of clusin, hinokinin and yatein. In one embodiment, the pharmaceutical composition comprises dihydrocubebin and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of dihydrocubebin, cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. One or both of dihydrocubebin and cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, dihydrocubebin may be present in a concentration of at least 5% (w/w), e.g. in a concentration of between 5% and 20% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25% (w/w), such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention. Furthermore, the present invention provides a pharmaceutical composition comprising hinokinin and yatein and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising hinokinin and yatein further comprises a compound selected from the group consisting of clusin, dihydrocubebin and cubebin. In one embodiment, the pharmaceutical composition comprises hinokinin and yatein as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of hinokinin, yatein and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. One or both of hinokinin and yatein may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, hinokinin may be present in a concentration of at least 1 5% (w/w), e.g. in a concentration of between 1 5% and 50% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2% (w/w), such as in a concentration of between 2% and 15% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising hinokinin and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising hinokinin and cubebin further comprises a compound selected from the group consisting of clusin, dihydrocubebin and yatein. In one embodiment, the pharmaceutical composition comprises hinokinin and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of hinokinin, cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. One or both of hinokinin and cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, hinokinin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 15% and 50% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25% (w/w), such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising yatein and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a preferred embodiment, the pharmaceutical composition comprising yatein and cubebin further comprises a compound selected from the group consisting of clusin, dihydrocubebin and hinokinin. In one embodiment, the pharmaceutical composition comprises yatein and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of yatein, cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. One or both of yatein and cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, yatein may be present in a concentration of at least 2% (w/w), e.g. in a concentration of between 2% and 15% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25% (w/w), such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention.

In a further preferred embodiment, the pharmaceutical composition according to the present invention comprises at least three compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, preferably the pharmaceutical composition comprises clusin, dihydrocubebin and hinokinin.

In a further preferred embodiment, the pharmaceutical composition according to the present invention comprises at least three compounds selected from the group consisting of clusin, dihydrocubebin, yatein and cubebin, preferably the pharmaceutical composition comprises clusin, dihydrocubebin and yatein. Any combination of the lignan compounds as disclosed herein may be comprised in the pharmaceutical composition according to the present invention. Thus, for example, the present invention provides a pharmaceutical composition comprising clusin, dihydrocubebin and hinokinin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises clusin, dihydrocubebin and hinokinin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, dihydrocubebin, hinokinin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Clusin, dihydrocubebin and/or hinokinin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or dihydrocubebin may be present in a concentration of, e.g. at least 5% (w/w), such as in a concentration of between 5% and 20% (w/w), and/or hinokinin may be present in a concentration of, e.g. at least 1 5%, such as in a concentration of between 15% and 50% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising clusin, dihydrocubebin and yatein and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises clusin, dihydrocubebin and yatein as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, dihydrocubebin, yatein and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Clusin, dihydrocubebin and/or yatein may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or dihydrocubebin may be present in a concentration of, e.g. at least 5% (w/w), such as in a concentration of between 5% and 20% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2%, such as in a concentration of between 2% and 1 5% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising clusin, dihydrocubebin and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises clusin, dihydrocubebin and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, dihydrocubebin, cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Clusin, dihydrocubebin and/or cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or dihydrocubebin may be present in a concentration of, e.g. at least 5% (w/w), such as in a concentration of between 5% and 20% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25%, such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising clusin, hinokinin and yatein and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises clusin, hinokinin and yatein as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, hinokinin, yatein and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Clusin, hinokinin and/or yatein may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or hinokinin may be present in a concentration of at least 15%, such as in a concentration of between 1 5% and 50% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2% (w/w), such as in a concentration of between 2% and 15% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising clusin, hinokinin and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises clusin, hinokinin and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, hinokinin, cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Clusin, hinokinin and/or cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or hinokinin may be present in a concentration of at least 15%, e.g. in a concentration of between 1 5% and 50% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25% (w/w), such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention. Furthermore, the present invention provides a pharmaceutical composition comprising clusin, yatein and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises clusin, yatein and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, yatein and cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Clusin, yatein and/or cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2% (w/w), such as in a concentration of between 2% and 1 5% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25%, such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising dihydrocubebin, hinokinin and yatein and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises dihydrocubebin, hinokinin and yatein as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of dihydrocubebin, hinokinin, yatein and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Dihydrocubebin, hinokinin and/or yatein may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, dihydrocubebin may be present in a concentration of at least 5% (w/w), e.g. in a concentration of between 5% and 20% (w/w), and/or hinokinin may be present in a concentration of, e.g. at least 15% (w/w), such as in a concentration of between 15% and 50% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2%, such as in a concentration of between 2% and 15% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising dihydrocubebin, hinokinin and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises dihydrocubebin, hinokinin and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of dihydrocubebin, hinokinin, cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Dihydrocubebin, hinokinin and/or cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, dihydrocubebin may be present in a concentration of at least 5% (w/w), e.g. in a concentration of between 5% and 20% (w/w), and/or hinokinin may be present in a concentration of, e.g. at least 15% (w/w), such as in a concentration of between 15% and 50% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25%, such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising dihydrocubebin, yatein and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises dihydrocubebin, yatein and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of dihydrocubebin, yatein, cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Dihydrocubebin, yatein and/or cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, dihydrocubebin may be present in a concentration of at least 5% (w/w), e.g. in a concentration of between 5% and 20% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2% (w/w), such as in a concentration of between 2% and 15% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25%, such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising hinokinin, yatein and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises hinokinin, yatein and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of hinokinin, yatein, cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Hinokinin, yatein and/or cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, hinokinin may be present in a concentration of at least 15% (w/w), e.g. in a concentration of between 1 5% and 50% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2%, such as in a concentration of between 2% and 1 5% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25% (w/w), such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention. Furthermore, the present invention provides a pharmaceutical composition comprising at least three compounds selected from the group consiting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, optionally one or two additional compounds selecetd from that group, such that the pharmaceutical composition according to the present invention comprises at least 3, optionally 4 or all of the compounds from the above group. Accordingly, the pharmaceutical composition of the present invention comprises clusin, dihydrocubebin and hinokinin, or dihydrokinin, hinokinin and yatein, or hinokinin, yatein and cubein, or yatein, cubebin and clusin, or cubebin, clusin and dihydrocubebin, or cubebin, dihydrocubebin and hinokinin, or yatein, cubebin, dihydrocubebin, or optionally the pharmacutical composition according to the present invention comprises one further compound. Thus, the pharmaceutical composition of the present invention may comprise clusin, dihydrocubebin, hinokinin and cubebin, or clusin, dihydrocubebin, hinokinin and yatein, or dihydrocubebin, hinokinin, yatein and cubebin, or dihydrocubebin, hinokinin, yatein and clusin, or hinokinin, yatein, cubein and clusin, or hinokinin, yatein, cubein and dihydrocubebin, or all of the compounds selected from the group, i.e. clusin, dihydrocubebin, hinokinin, yatein and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Clusin, dihydrocubebin, hinokinin, yatein and cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or dihydrocubebin may be present in a concentration of, e.g. at least 5% (w/w), such as in a concentration of between 5% and 20% (w/w), and/or hinokinin may be present in a concentration of, e.g. at least 1 5%, such as in a concentration of between 15% and 50% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2% (w/w), such as in a concentration of between 2% and 1 5% (w/w), ), and/or cubebin may be present in a concentration of, e.g. at least 25%, such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention.

In a further preferred embodiment, the pharmaceutical composition according to the present invention comprises at least four compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Any combination of the lignan compounds as disclosed herein may be comprised in the pharmaceutical composition according to the present invention.

Thus, for example, the present invention provides a pharmaceutical composition comprising clusin, dihydrocubebin, hinokinin and yatein and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises clusin, dihydrocubebin, hinokinin and yatein as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, dihydrocubebin, hinokinin, yatein and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Clusin, dihydrocubebin, hinokinin and/or yatein may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or dihydrocubebin may be present in a concentration of, e.g. at least 5% (w/w), such as in a concentration of between 5% and 20% (w/w), and/or hinokinin may be present in a concentration of, e.g. at least 1 5%, such as in a concentration of between 15% and 50% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2% (w/w), such as in a concentration of between 2% and 15% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising clusin, dihydrocubebin, hinokinin and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises clusin, dihydrocubebin, hinokinin and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, dihydrocubebin, hinokinin, cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Clusin, dihydrocubebin, hinokinin and/or cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or dihydrocubebin may be present in a concentration of, e.g. at least 5% (w/w), such as in a concentration of between 5% and 20% (w/w), and/or hinokinin may be present in a concentration of, e.g. at least 1 5%, such as in a concentration of between 15% and 50% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25% (w/w), such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising clusin, hinokinin, yatein and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises clusin, hinokinin, yatein and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, hinokinin, yatein, cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Clusin, hinokinin, yatein and/or cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or hinokinin may be present in a concentration of, e.g. at least 15%, such as in a concentration of between 1 5% and 50% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2% (w/w), such as in a concentration of between 2% and 15% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25% (w/w), such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention. Furthermore, the present invention provides a pharmaceutical composition comprising clusin, dihydrocubebin, yatein and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises clusin, dihydrocubebin, yatein and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of clusin, dihydrocubebin, yatein, cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Clusin, dihydrocubebin, yatein and/or cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or dihydrocubebin may be present in a concentration of at least 5% (w/w), e.g. in a concentration of between 5% and 20% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2%, such as in a concentration of between 2% and 1 5% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25% (w/w), such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention.

Furthermore, the present invention provides a pharmaceutical composition comprising dihydrocubebin, hinokinin, yatein and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. In one embodiment, the pharmaceutical composition comprises dihydrocubebin, hinokinin, yatein and cubebin as the only pharmaceutically active ingredients. In one embodiment, the pharmaceutical composition according the present invention consists of dihydrocubebin, hinokinin, yatein, cubebin and one or more pharmaceutically acceptable excipients, carriers, and/or diluents. Dihydrocubebin, hinokinin, yatein and/or cubebin may be present in the pharmaceutical composition according to the present invention, e.g. in a concentration as disclosed above for the pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, dihydrocubebin may be present in a concentration of at least 5% (w/w), e.g. in a concentration of between 5% and 20% (w/w), and/or hinokinin may be present in a concentration of, e.g. at least 1 5% (w/w), such as in a concentration of between 1 5% and 50% (w/w), and/or may be present in a concentration of, e.g. at least 2%, such as in a concentration of between 2% and 15% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25% (w/w), such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention.

In one embodiment, the pharmaceutical composition according to the present invention comprises at least one, preferably at least two, but not more than four, i.e. two, three or four, compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, the present invention provides, a pharmaceutical composition comprising at least two, but not more than four, i.e. two, three or four, compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer.

For example, clusin may be absent from the pharmaceutical composition according to the present invention, or dihydrocubebin may be absent from the pharmaceutical composition according to the present invention, or hinokinin may be absent from the pharmaceutical composition according to the present invention, or yatein may be absent from the pharmaceutical composition according to the present invention, or cubebin may be absent from the pharmaceutical composition according to the present invention. Any combination of the lignan compounds as disclosed herein may be comprised in the pharmaceutical composition according to this embodiment of the present invention, except of the combination clusin, dihydrocubebin, hinokinin, yatein and cubebin.

For example, hinokinin and cubebin may be absent from the pharmaceutical composition according to the present invention, or clusin and hinokinin may be absent from the pharmaceutical composition according to the present invention, or yatein and cubebin may be absent from the pharmaceutical composition according to the present invention, dihydrocubebin and yatein may be absent from the pharmaceutical composition according to the present invention, or hinokinin and yatein may be absent from the pharmaceutical composition according to the present invention, or dihydrokinin and hinokinin may be absent from the pharmaceutical composition according to the present invention, or clusin and yatein may be absent from the pharmaceutical composition according to the present invention, or clusin and dihydrocubebin may be absent from the pharmaceutical composition according to the present invention, or clusin and cubebin may be absent from the pharmaceutical composition according to the present invention, or diydrocubebin and cubebin may be absent from the pharmaceutical composition according to the present invention.

Any combination of the lignan compounds as disclosed herein may be comprised in the pharmaceutical composition according to this embodiment of the present invention, except of the combination clusin, dihydrocubebin, hinokinin, yatein and cubebin. For example, the pharmaceutical composition comprising at least two, but not more than four compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin may comprise or consist of the combinations of lignan compounds, including the preferred concentrations of the lignan compounds, as defined above for pharmaceutical compositions comprising or consisting of two, three or four lignan compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin.

Accordingly, in a preferred embodiment the pharmaceutical composition of the present invention provides a pharmaceutical composition which preferably comprises yatein and dihydrocubebin and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, such as hormone-dependent cancer, e.g. hormone- dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone- dependent prostate cancer, wherein

yatein is present in a concentration of at least 2% (w/w), such as at least 2% (w/w), at least 4% (w/w), at least 8% (w/w), at least 15% (w/w), at least 30% (w/w), or at least 50% (w/w), e.g. in a concentration of between about 2% and 50% (w/w), such as in a concentration of between about 2% and 15% (w/w), and

dihydrocubebin is present in a concentration of at least 5% (w/w), such as at least 5% (w/w), at least 10% (w/w), at least 1 5% (w/w), at least 20% (w/w), at least 25% (w/w), or at least 30% (w/w), more preferably in a concentration of between about 5% and 50% (w/w), most preferably in a concentration of between about 5% and 25% (w/w).

In a particularly preferred embodiment the pharmaceutical composition of the present invention provides a pharmaceutical composition which preferably comprises clusin and yatein and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, such as hormone-dependent cancer, e.g. hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone- dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer, wherein

clusin is present in a concentration of at least 10% (w/w), such as at least 10% (w/w), at least 20% (w/w), at least 30% (w/w), at least 40% (w/w), at least 50% (w/w), or at least 60% (w/w), e.g. in a concentration of between about 10% and 60% (w/w), such as in a concentration of between about 10% and 30% (w/w),

yatein is present in a concentration of at least 2% (w/w), such as at least 2% (w/w), at least 4% (w/w), at least 8% (w/w), at least 15% (w/w), at least 30% (w/w), or at least 50% (w/w), e.g. in a concentration of between about 2% and 50% (w/w), such as in a concentration of between about 2% and 15% (w/w).

Thus, in one particularly preferred embodiment, the pharmaceutical composition of the present invention provides a pharmaceutical composition which preferably comprises clusin, dihydrocubebin and yatein, or clusin, dihydrocubebin and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, such as hormone-dependent cancer, e.g. hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer, wherein

dihydrocubebin is present in a concentration of at least 5% (w/w), such as at least 5% (w/w), at least 10% (w/w), at least 15% (w/w), at least 20% (w/w), at least 25% (w/w), or at least 30% (w/w), more preferably in a concentration of between about 5% and 50% (w/w), most preferably in a concentration of between about 5% and 25% (w/w),

cubebin is present in a concentration of at least 25% (w/w), such as at least 25% (w/w), at least 35% (w/w), at least 45% (w/w), at least 55% (w/w), at least 65% (w/w), or at least 75% (w/w), e.g. in a concentration of between about 25% and 80% (w/w), such as in a concentration of between about 25% and 60% (w/w).

Thus, in one preferred embodiment, the pharmaceutical composition of the present invention provides a pharmaceutical composition which preferably comprises clusin, dihydrocubebin, yatein and hinokinin and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, such as hormone-dependent cancer, e.g. hormone- dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone- dependent prostate cancer, wherein

yatein is present in a concentration of at least 2% (w/w), such as at least 2% (w/w), at least 4% (w/w), at least 8% (w/w), at least 15% (w/w), at least 30% (w/w), or at least 50% (w/w), e.g. in a concentration of between about 2% and 50% (w/w), such as in a concentration of between about 2% and 1 5% (w/w), and

hinokinin is present in a concentration of at least 15% (w/w), such as at least 15% (w/w), at least 20% (w/w), at least 30% (w/w), at least 40% (w/w), at least 50% (w/w), or at least 60% (w/w), more preferably in a concentration of between about 15% and 70% (w/w), most preferably in a concentration of between about 15% and 50% (w/w).

In the above pharmaceutical compositions comprising two, three or four of the lignan compounds according to the present invention, the ratios of the lignans within the pharmaceutical compositions may differ from the ratios of the Piper cubeba extracts according to the present invention, e.g. to the Piper cubeba fraction according to Example 1 1 (Pip.cub.40p2.001 .E03.S04). Accordingly, the compositions may comprise more or less of any one of the compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin than the Piper cubeba fraction according to the present invention, i.e. Pip.cub.40p2.001 .E03.S04. However, the ratios of the lignans in the pharmaceutical composition according to the invention may correspond to the ratios of the compounds in the inventive Piper cubeba fraction Pip.cub.40p2.001 .E03.S04.

In one embodiment, the pharmaceutical composition according the present invention comprises clusin, dihydrocubebin, hinokinin, yatein and cubebin, and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, such as hormone-dependent cancer, e.g. hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Preferred concentrations of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin are concentrations, e.g. as defined above for the pharmaceutical compositions comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w) as described above, e.g. in a concentration of between 10% and 30% (w/w), and/or dihydrocubebin may be present in a concentration of, e.g. at least 5% (w/w) as described above, e.g. in a concentration of between 5% and 20% (w/w), and/or hinokinin may be present in a concentration of, e.g. at least 1 5% (w/w) as described above, e.g. in a concentration of between 15% and 50% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2% (w/w) as described above, e.g. in a concentration of between 2% and 1 5% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25% (w/w) as described above, e.g. in a concentration of between 25% and 60% (w/w).

Thus, in a particularly preferred embodiment, the present invention provides a pharmaceutical composition comprising clusin, dihydrocubebin, hinokinin, yatein and cubebin, and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer, wherein

hinokinin is present in a concentration of at least 15% (w/w), such as at least 1 5% (w/w), at least 20% (w/w), at least 30% (w/w), at least 40% (w/w), at least 50% (w/w), or at least 60% (w/w), more preferably in a concentration of between about 15% and 70% (w/w), most preferably in a concentration of between about 15% and 50% (w/w),

In the above pharmaceutical compositions comprising clusin, dihydrocubebin, hinokinin, yatein and cubebin the ratios of the lignans within the pharmaceutical compositions may differ from the ratios of the Piper cubeba extracts according to the present invention, e.g. to the Piper cubeba extract according to Example 1 1 (Pip. Cub. 40p2.001 .E03.S04). Accordingly, the compositions may comprise more or less of any one of the compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin than the Piper cubeba extract according to the present invention, i.e. Pip. Cub. 40p2.001 .E03.S04. However, the ratios of the lignans in the pharmaceutical composition according to the invention may correspond to the ratios of the compounds in the inventive Piper cubeba extract Pip. Cub. 40p2.001 .E03.S04.

In a preferred embodiment of the pharmaceutical composition according to the present invention, the concentration of lignan compounds together, preferably the concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together is at least 60% (w/w), preferably at least 65% (w/w), more preferably at least 70% (w/w), even more preferably at least 75% (w/w), and even more preferably at least 80%, and may be up to 100%, such as 90% (w/w), 95% (w/w), 99% (w/w) or 100%. Thus, the present invention provides a pharmaceutical composition comprising at least one, preferably at least two, such as two, three, four or five compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, preferably selected from the group consisting of clusin, dihydrocubebin and hinokinin, more preferably selected from the group consisting of clusin and dihydrocubebin, and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents as disclosed herein, wherein the concentration of lignan compounds together, preferably the concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together is at least 60% (w/w), preferably at least 65% (w/w), more preferably at least 70% (w/w), even more preferably at least 75% (w/w), and even more preferably at least 80% or higher, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, such as hormone-dependent cancer, e.g. hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone- dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer.

Furthermore, the present invention provides a pharmaceutical composition comprising at least one, preferably at least two, but not more than four, i.e. two, three or four compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents as disclosed above, wherein preferably the concentration of lignan compounds together, preferably the concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together is at least 60% (w/w), preferably at least 65% (w/w), more preferably at least 70% (w/w), and even more preferably at least 75% (w/w) and may be up to 100%, preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, such as hormone-dependent cancer, e.g. hormone- dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone- dependent prostate cancer. For example, the pharmaceutical composition according to the present invention may comprise clusin in a concentration of at least 10% as described above and may comprise a total lignan concentration, preferably a lignan concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together, of at least 60%, preferably at least 65% (w/w), more preferably at least 70% (w/w), and even more preferably at least 75% (w/w), and/or the pharmaceutical composition according to the present invention may comprise dihydrocubebin in a concentration of at least 5% as described above and may comprise a total lignan concentration, preferably a lignan concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together, of at least 60%, preferably at least 65% (w/w), more preferably at least 70% (w/w), and even more preferably at least 75% (w/w), and/or the pharmaceutical composition according to the present invention may comprise hinokinin in a concentration of at least 15% as described above and may comprise a total lignan concentration, preferably a lignan concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together, of at least 60%, preferably at least 65% (w/w), more preferably at least 70% (w/w), and even more preferably at least 75% (w/w), and/or the pharmaceutical composition according to the present invention may comprise yatein in a concentration of at least 2% as described above and may comprise a total lignan concentration, preferably a lignan concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together, of at least 60%, preferably at least 65% (w/w), more preferably at least 70% (w/w), and even more preferably at least 75% (w/w), and/or the pharmaceutical composition according to the present invention may comprise cubebin in a concentration of at least 25% as described above and may comprise a total lignan concentration, preferably a lignan concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together, of at least 60%, preferably at least 65% (w/w), more preferably at least 70% (w/w), and even more preferably at least 75% (w/w). In a particularly preferred embodiment, the present invention provides a pharmaceutical composition comprising clusin and dihydrocubebin, preferably comprising clusin, dihydrocubebin and hinokinin, more preferably comprising clusin, dihydrocubebin, hinokinin, yatein and cubebin, and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents, wherein the concentration of lignan compounds together, preferably the concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together, is at least 60% (w/w), preferably at least 65% (w/w), more preferably at least 70% (w/w), and even more preferably at least 75% (w/w), preferably for use as a medicament, more preferably for use in the treatment or prevention of a tumour disease as described herein, even more preferably for use in the treatment or prevention of cancer, such as hormone- dependent cancer, e.g. hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone- dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer, such as hormone-dependent prostate cancer.

In one embodiment, the pharmaceutical composition according to the present invention comprises at least two compounds selected from the group consisting of clusin, cubebin, dihydrocubebin, hinokinin and yatein, wherein at least one of the compounds is present in the following concentrations: clusin in a concentration of at least 10% (w/w), such as at least 10% (w/w), at least 20% (w/w), at least 30% (w/w), at least 40% (w/w), at least 50% (w/w), or at least 60% (w/w), e.g. in a concentration of between about 10% and 60% (w/w), such as in a concentration of between about 10% and 30% (w/w),

dihydrocubebin in a concentration of at least 5% (w/w), such as at least 5% (w/w), at least 10% (w/w), at least 15% (w/w), at least 20% (w/w), at least 25% (w/w), or at least 30% (w/w), e.g. in a concentration of between about 5% and 50% (w/w), such as in a concentration of between about 5% and 25% (w/w),

- hinokinin in a concentration of at least 15% (w/w), such as at least 15% (w/w), at least 20% (w/w), at least 30% (w/w), at least 40% (w/w), at least 50% (w/w), or at least 60% (w/w), e.g. in a concentration of between about 15% and 70% (w/w), such as in a concentration of between about 15% and 50% (w/w),

yatein in a concentration of at least 2% (w/w), such as at least 2% (w/w), at least 4% (w/w), at least 8% (w/w), at least 1 5% (w/w), at least 30% (w/w), or at least 50% (w/w), e.g. in a concentration of between about 2% and 50% (w/w), such as in a concentration of between about 2% and 15% (w/w),

cubebin in a concentration of at least 25% (w/w), such as at least 25% (w/w), at least 35% (w/w), at least 45% (w/w), at least 55% (w/w), at least 65% (w/w), or at least 75% (w/w), more preferably in a concentration of between about 25% and 80% (w/w), most preferably in a concentration of between about 25% and 60% (w/w).

Thus, for example, in one embodiment, the pharmaceutical composition according to the present invention comprises clusin, wherein clusin is present in a concentration of at least 10% (w/w) as described above, preferably for the use as described above. Preferably, the pharmaceutical composition comprising clusin, wherein clusin is present in a concentration of at least 10% (w/w), further comprises at least one further compound selected from the group consisting of dihydrocubebin, cubebin, hinokinin and yatein, preferably at least two further compounds selected from the group consisting of dihydrocubebin, cubebin, hinokinin and yatein, preferably at least three further compounds selected from the group consisting of dihydrocubebin, cubebin, hinokinin and yatein. Any of the above defined combinations of lignan compounds comprising clusin are encompassed for this embodiment of the present invention, wherein clusin is present in a concentration of at least 10% (w/w). Preferably, the pharmaceutical composition comprising clusin further comprises dihydrocubebin, preferably in a concentration of at least 5% (w/w), more preferably in a concentration of between 5% and 20% (w/w), and/or hinokinin, preferably in a concentration of at least 1 5% (w/w), more preferably in a concentration of between 15% and 50% (w/w), and/or yatein, preferably in a concentration of at least 2% (w/w), more preferably in a concentration of between 2% and 1 5% (w/w), and/or cubebin, preferably in a concentration of at least 25% (w/w), more preferably in a concentration of between 25% and 60% (w/w).

Furthermore, in another exemplary embodiment, the pharmaceutical composition according to the present invention comprises dihydrocubebin, wherein dihydrocubebin is present in a concentration of at least 5% (w/w) as described above, preferably for the use as described above. Preferably, the pharmaceutical composition comprising dihydrocubebin, wherein dihydrocubebin is present in a concentration of at least 5% (w/w), further comprises at least one further compound selected from the group consisting of clusin, cubebin, hinokinin and yatein, preferably at least two further compounds selected from the group consisting of clusin, cubebin, hinokinin and yatein, preferably at least three further compounds selected from the group consisting of clusin, cubebin, hinokinin and yatein. Any of the above defined combinations of lignan compounds comprising dihydrocubebin are encompassed for this embodiment of the present invention, wherein dihydrocubebin is present in a concentration of at least 5% (w/w). Preferably, the pharmaceutical composition comprising dihydrocubebin further comprises clusin, preferably in a concentration of at least 10% (w/w), more preferably in a concentration of between 10% and 30% (w/w), and/or hinokinin, preferably in a concentration of at least 15% (w/w), more preferably in a concentration of between 15% and 50% (w/w), and/or yatein, preferably in a concentration of at least 2% (w/w), more preferably in a concentration of between 2% and 15% (w/w), and/or cubebin, preferably in a concentration of at least 25% (w/w), more preferably in a concentration of between 25% and 60% (w/w). Furthermore, in another exemplary embodiment, the pharmaceutical composition according to the present invention comprises hinokinin, wherein hinokinin is present in a concentration of at least 15% (w/w) as described above, preferably for the use as described above. Preferably, the pharmaceutical composition comprising hinokinin, wherein hinokinin is present in a concentration of at least 1 5% (w/w), further comprises at least one further compound selected from the group consisting of clusin, cubebin, dihydrocubebin and yatein, preferably at least two further compounds selected from the group consisting of clusin, cubebin, dihydrocubebin and yatein, preferably at least three further compounds selected from the group consisting of clusin, cubebin, dihydrocubebin and yatein. Any of the above defined combinations of lignan compounds comprising hinokinin are encompassed for this embodiment of the present invention, wherein hinokinin is present in a concentration of at least 15% (w/w). Preferably, the pharmaceutical composition comprising hinokinin further comprises clusin, preferably in a concentration of at least 10% (w/w), more preferably in a concentration of between 10% and 30% (w/w), and/or dihydrocubebin, preferably in a concentration of at least 5% (w/w), more preferably in a concentration of between 5% and 20% (w/w), and/or yatein, preferably in a concentration of at least 2% (w/w), more preferably in a concentration of between 2% and 15% (w/w), and/or cubebin, preferably in a concentration of at least 25% (w/w), more preferably in a concentration of between 25% and 60% (w/w).

Furthermore, in another exemplary embodiment, the pharmaceutical composition according to the present invention comprises yatein, wherein yatein is present in a concentration of at least 2% (w/w) as described above, preferably for the use as described above. Preferably, the pharmaceutical composition comprising yatein, wherein yatein is present in a concentration of at least 2% (w/w), further comprises at least one further compound selected from the group consisting of clusin, cubebin, hinokinin and dihydrocubebin, preferably at least two further compounds selected from the group consisting of clusin, cubebin, hinokinin and dihydrocubebin, preferably at least three further compounds selected from the group consisting of clusin, cubebin, hinokinin and dihydrocubebin. Any of the above defined combinations of lignan compounds comprising yatein are encompassed for this embodiment of the present invention, wherein yatein is present in a concentration of at least 2% (w/w). Preferably, the pharmaceutical composition comprising yatein further comprises clusin, preferably in a concentration of at least 10% (w/w), more preferably in a concentration of between 10% and 30% (w/w), and/or hinokinin, preferably in a concentration of at least 15% (w/w), more preferably in a concentration of between 15% and 50% (w/w), and/or dihydrocubebin, preferably in a concentration of at least 5% (w/w), more preferably in a concentration of between 5% and 20% (w/w), and/or cubebin, preferably in a concentration of at least 25% (w/w), more preferably in a concentration of between 25% and 60% (w/w).

Furthermore, in another exemplary embodiment, the pharmaceutical composition according to the present invention comprises cubebin, wherein cubebin is present in a concentration of at least 25% (w/w) as described above, preferably for the use as described above. Preferably, the pharmaceutical composition comprising cubebin, wherein cubebin is present in a concentration of at least 25% (w/w), further comprises at least one further compound selected from the group consisting of clusin, dihydrocubebin, hinokinin and yatein, preferably at least two further compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin and yatein, preferably at least three further compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin and yatein. Any of the above defined combinations of lignan compounds comprising cubebin are encompassed for this embodiment of the present invention, wherein cubebin is present in a concentration of at least 25% (w/w). Preferably, the pharmaceutical composition comprising cubebin further comprises clusin, preferably in a concentration of at least 10% (w/w), more preferably in a concentration of between 10% and 30% (w/w), and/or hinokinin, preferably in a concentration of at least 1 5% (w/w), more preferably in a concentration of between 1 5% and 50% (w/w), and/or yatein, preferably in a concentration of at least 2% (w/w), more preferably in a concentration of between 2% and 15% (w/w), and/or dihydrocubebin, preferably in a concentration of at least 5% (w/w), more preferably in a concentration of between 5% and 20% (w/w).

In a preferred embodiment, the pharmaceutical composition according to the present invention does not comprise proteins in a significant amount, preferably the pharmaceutical composition according to the present invention is free of proteins. In another preferred embodiment, the pharmaceutical composition according to the present invention does not comprise amino acids in a significant amount, preferably the pharmaceutical composition according to the present invention is free of amino acids. Thus, for example, the pharmaceutical composition according to the present invention is essentially free from proteins and/or amino acids, preferably the pharmaceutical composition according to the present invention does not comprise proteins and/or amino acids. The term "significant amount" preferably means in the context of the present invention an amount of more than 0.5% (w/w). Thus, for example, the pharmaceutical composition according to the present invention, preferably does not comprise more than 0.5% (w/w) proteins and/or preferably does not comprise more than 0.5% (w/w) amino acids. In the context of the present invention, the term "essentially free" preferably refers to an amount of at most 0.5% (w/w), more preferably of at most 0.3% (w/w), even more preferably of at most 0.1 % (w/w), and most preferably to an amount of below the detection limit (using state of the art detection methods for the respective compounds). Furthermore, in one embodiment, the pharmaceutical composition according to the present invention is not an extract type of Piper cubeba as disclosed in WO 2009/021347 or as disclosed by Yam et al., Planta Med. 2008, 74:33-38. Thus, for example, the pharmaceutical composition according to the present invention is not an extract from Piper cubeba prepared by ethanol extraction of CO₂ defatted powder of Piper cubeba using ethanol absolute or a dilution of ethanol, such as 90% ethanol aq. (w/w), stirring for 2 hours at room temperature, and removing solid plant material from the obtained extract, such as by filtration, e.g. using a cellulose filter AF6, without any further purification steps.

Furthermore, in one embodiment, the pharmaceutical composition according to the invention is not an extract of Piper cubeba as disclosed by Elfami et al., Biochemical Systematics and Ecology 35 (2007) 397 - 402. The pharmaceutical composition according to the present invention is thus not an extract of dried Piper cubeba ground with quarz, extracted under sonification for 1 h using 2ml of 80% methanol, followed by fractionation using 4,0ml of CH₂CI₂ and 4ml of water and then subsequently shaken for 5sec and centrifuged at 1500g for 5 minutes, followed by, e.g. evaporation and preparative thin layer chromatography (TLC).

Furthermore, in one embodiment, the pharmaceutical composition according to the present invention does not comprise a dichloromethane extract of Pycanthus angolensis bark, such as e.g. hinokinin or a compound obtained by reduction of said extract as disclosed by Pusztai et al., Anticancer Research 30: 451 -454 (2010), such as e.g. (-)-dihydrocubebin.

Further, in one embodiment of the invention, the pharmaceutical composition according to the invention is not an extract type of Piper cubeba as disclosed in Bharathi et al., Phytochemistry, Vol. 24, No. 2, pp. 329 - 331 , 1985. Thus, for example, the pharmaceutical composition according to the present invention is not an extract from Piper cubeba prepared by petrol extraction, e.g. from ground P. cubeba fruits extracted with petrol for 56h and subsequent solvent removal, e.g. under reduced pressure, followed by e.g. trituration with ethanol, reduction in vacuo, followed by, e.g. repeated column chromatography over neutral Al₂0₃ using solvents with increasing polarity from petrol to ethyl acetate, followed by e.g. preparative thin layer chromatography.

In a further aspect, the present invention provides clusin, dihydrocubebin, or clusin, yatein, or clusin, dihydrocubebin and yatein for use in the treatment or prevention of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, such as hormone- dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone- dependent prostate cancer. Preferred tumour diseases are selected from prostate cancer, breast cancer and pancreatic cancer. The compounds clusin, dihydrocubebin and/or yatein may be used alone or in combination. If used in combination, the compounds may be used together in a pharmaceutical composition as disclosed above or separately, for example, in separate pharmaceutical compositions, e.g. for concurrent or sequential administration and/or for the same or different administration routes.

Thus, in one embodiment, the present invention provides clusin for use in the treatment or prevention of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone- dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides dihydrocubebin for the treatment of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides yatein for use in the treatment of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. For this aspect of the present invention, clusin, dihydrocubebin and yatein are as described throughout the specification of the present invention. Furthermore, the disclosure provided for the pharmaceutical composition according to the present invention, e.g. the concentrations and combinations of two or three lignan compounds selected from clusin, dihydrocubebin and yatein also applies to this aspect of the present invention.

In a further aspect, the present invention provides clusin, dihydrocubebin and/or hinokinin for use in the treatment or prevention of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, such as hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Preferred tumour diseases are selected from prostate cancer, breast cancer and pancreatic cancer. The compounds clusin, dihydrocubebin and/or hinokinin may be used alone or in combination. If used in combination, the compounds may be used together in a pharmaceutical composition as disclosed above or separately, for example, in separate pharmaceutical compositions, e.g. for concurrent or sequential administration and/or for the same or different administration routes.

Thus, in one embodiment, the present invention provides clusin for use in the treatment or prevention of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone- dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides dihydrocubebin for the treatment of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides hinokinin for use in the treatment of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer.

For this aspect of the present invention, clusin, dihydrocubebin and hinokinin are as described throughout the specification of the present invention. Furthermore, the disclosure provided for the pharmaceutical composition according to the present invention, e.g. the concentrations and combinations of two or three lignan compounds selected from clusin, dihydrocubebin and hinokinin also applies to this aspect of the present invention.

In a further aspect, the present invention provides clusin, dihydrocubebin, or clusin, cubebin, or clusin, dihydrocubebin and cubebin for use in the treatment or prevention of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, such as hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone- dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone- dependent prostate cancer. Preferred tumour diseases are selected from prostate cancer, breast cancer and pancreatic cancer. The compounds clusin, dihydrocubebin and/or cubebin may be used alone or in combination. If used in combination, the compounds may be used together in a pharmaceutical composition as disclosed above or separately, for example, in separate pharmaceutical compositions, e.g. for concurrent or sequential administration and/or for the same or different administration routes.

Thus, in the above embodiment, the present invention provides clusin for use in the treatment or prevention of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone- dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides dihydrocubebin for the treatment of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides cubebin for use in the treatment of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer.

For this aspect of the present invention, clusin, dihydrocubebin and cubebin are as described throughout the specification of the present invention. Furthermore, the disclosure provided for the pharmaceutical composition according to the present invention, e.g. the concentrations and combinations of two or three lignan compounds selected from clusin, dihydrocubebin and cubebin also applies to this aspect of the present invention.

In a further aspect, the present invention provides clusin, dihydrocubebin, yatein and hinokinin for use in the treatment or prevention of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, such as hormone-dependent breast cancer, hormone- dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Preferred tumour diseases are selected from prostate cancer, breast cancer and pancreatic cancer. The compounds clusin, dihydrocubebin and/or yatein may be used alone or in combination. If used in combination, the compounds may be used together in a pharmaceutical composition as disclosed above or separately, for example, in separate pharmaceutical compositions, e.g. for concurrent or sequential administration and/or for the same or different administration routes. Thus, in one embodiment, the present invention provides clusin for use in the treatment or prevention of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone- dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides dihydrocubebin for the treatment of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides yatein for use in the treatment of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides hinokinin for use in the treatment of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. For this aspect of the present invention, clusin, dihydrocubebin, yatein and hinokin are as described throughout the specification of the present invention. Furthermore, the disclosure provided for the pharmaceutical composition according to the present invention, e.g. the concentrations and combinations of two, three or four lignan compounds selected from clusin, dihydrocubebin, yatein and hinokinin also applies to this aspect of the present invention.

In a further aspect, the present invention provides clusin, dihydrocubebin, yatein, hinokinin and cubein for use in the treatment or prevention of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, such as hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone- dependent colorectal cancer, and hormone-dependent bladder cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Preferred tumour diseases are selected from prostate cancer, breast cancer and pancreatic cancer. The compounds clusin, dihydrocubebin and/or yatein may be used alone or in combination. If used in combination, the compounds may be used together in a pharmaceutical composition as disclosed above or separately, for example, in separate pharmaceutical compositions, e.g. for concurrent or sequential administration and/or for the same or different administration routes. Thus, in one embodiment, the present invention provides clusin for use in the treatment or prevention of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone- dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides dihydrocubebin for the treatment of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides yatein for use in the treatment of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides hinokinin for use in the treatment of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides cubebin for the treatment of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer.

For this aspect of the present invention, clusin, dihydrocubebin, yatein, hinokinin and cubebin are as described throughout the specification of the present invention. Furthermore, the disclosure provided for the pharmaceutical composition according to the present invention, e.g. the concentrations and combinations of two, three, four or five lignan compounds selected from clusin, dihydrocubebin, yatein, hinokinin and cubebin also applies to this aspect of the present invention.

In a further aspect, the present invention provides a process for preparing a lignan composition comprising a lignan concentration of at least 60% (w/w), preferably of at least 65% (w/w), more preferably of at least 70% (w/w), even more preferably at least 75% (w/w), from lignan- containing plant material. The lignan composition prepared according to the process according to the present invention may have a lignan concentration of up to 100%, such as about 90% (w/w), about 95% (w/w), about 99% (w/w) or 100%. Preferably, the lignan composition obtainable according to the process according to the present invention comprises clusin, dihydrocubebin, hinokinin, yatein and cubebin. Preferably, the lignan concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together is at least 60% (w/w), preferably at least 65% (w/w), more preferably at least 70% (w/w), even more preferably at least 75% (w/w) and may be up to 100%, such as about 90% (w/w), about 95% (w/w), about 99% (w/w) or 100%, in the lignan composition obtainable by the process according to the present invention. The lignan concentration in the lignan composition may be determined by HPLC technology.

The concentrations of the single lignan compounds of the lignan composition obtainable according to the process according to the present invention are preferably as follows:

clusin is preferably present in a concentration of at least 10% (w/w), preferably between 10% and 30% (w/w),

dihydrocubebin is preferably present in a concentration of at least 5% (w/w), preferably between 5% and 20% (w/w),

hinokinin is preferably present in a concentration of at least 15% (w/w), preferably between 1 5% and 50% (w/w),

yatein is preferably present in a concentration of at least 2% (w/w), preferably between 2% and 15% (w/w), and

cubebin is preferably present in a concentration of at least 25% (w/w), preferably between 25% and 60% (w/w).

The lignan concentrations of the single lignan compounds in the lignan composition may be determined by HPLC technology.

The process according to the present invention comprises the steps of:

(a) contacting lignan-containing plant material with an extraction solvent to obtain an extraction solution,

(b) separating the plant material solids from said extraction solution,

(c) optionally concentrating said extraction solution,

(d) fractionating the extraction solution obtained in step b) or c),

(e) pooling lignan containing fractions, and (f) optionally concentrating the pooled lignan-containing fractions

to obtain a lignan composition, wherein the lignan concentration, preferably the lignan concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together, in the lignan composition is at least 60% (w/w), preferably at least 70% (w/w), more preferably at least 75% (w/w). Steps (d) to (f) may optionally be repeated for further purification.

The term "contacting" with an extraction solvent in the context of the process according to the present invention preferably refers to wetting, submerging, bathing or any other type of incubation of the plant material with an extraction solution.

The term "extraction solvent" in the context of the present invention refers to any solvent or liquid, which is suitable for the process disclosed herein. Suitable extraction solvents are known to a person skilled in the art and may include, but are not limited to water, supercritical gases, such as supercritical C0₂, organic solvents, alcohols, such as e.g. methanol, ethanol, isopropanol, aromatic alcohols and their aqueous dilutions, wherein the alcohol is present in an amount of e.g. 20% to 95,58% (v/v), preferably 30% to 99% (v/v), preferably 40 to 99% (v/v), preferably 50% to 99% (v/v), preferably 60% to 99% (v/v), preferably 70% to 99% (v/v), most preferably 80% to 99% (v/v). The extraction solvent may also be any of the organic solvents, such as for example diethyl ether, chloroform, 1 ,4-dioxane, toluene, benzene, cyclohexane, hexane, pentane, dichloromethane, tetrahydrofuran, ethyl acetate, acetone, dimethylformamide, acetonitrile, dimethyl sulfoxide, or any combination thereof. It will be apparent for the skilled person that the extraction solvent can also be a mixture of alcohol and another organic solvent, or a water-saturated alcohol/organic solvent solution. In a preferred embodiment of the process according to the present invention, the extraction solvent is ethanol or an aqueous dilution of ethanol, such as absolute ethanol or e.g. 80% to 99% ethanol aq. (v/v), such as 90% ethanol aq. (v/v). Furthermore, supercritical gases, such as supercritical C0₂, are possible and preferred extraction solvents.

The term "lignan composition" as used herein refers to a composition comprising one or more lignan compounds, such as clusin, dihydrocubebin, hinokinin, yatein and/or cubebin. The lignan composition obtainable by the process according to the present invention comprises lignan compounds, preferably the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin, in a total concentration of at least 60% (w/w), preferably in a total concentration of at least 65% (w/w), more preferably in a total concentration of at least 70% (w/w), even more preferably in a total concentration of at least 75% (w/w) and may comprise a total lignan concentration of up to 1 00%, such as about 90% (w/w), about 95% (w/w), about 99% (w/w) or 1 00%.

Preferably, the separation step (b) of the process according to the present invention may comprise filtration or centrifugation. Suitable filtration procedures or means of centrifugation are known to a person skilled in the art.

The term "fi ltration" as used herein refers to the mechanical or physical operation which is used for the separation of solids from fluids by interposing a medium through which only the fluid can pass. The term "centrifugation" as used herein also refers to differential centrifugation, isopycnic centrifugation, and ultracentrifugation. Various methods of centrifugation are known in prior art and are readily available.

The optional concentration step (c) of the process according to the invention preferably comprises evaporation or precipitation, more preferably evaporation. In a preferred embodiment of the present invention, step (c) involves or comprises evaporation. It wil l be evident to the skilled person that evaporation can be facilitated by elevated temperatures of, e.g. about 20°C to 200°C, preferably of about 20° to 1 00°C, preferably of about 30°C to 90°C, more preferably of about 40°C to 80°C, even more preferably of about 40°C to 70°C, such as of about 50°C to 60°C. The evaporation process may also be facilitated when carried out under reduced pressure of, e.g. I mbar to 500mbar, preferably l Ombar to 250mbar, preferably 20 to l OOmbar, preferably 24 to 70mbar, preferably 25mbar to 80mbar, preferably 30mbar to 90mbar, preferably at 10 to 10Ombar, preferably at 1 5mbar to 70mbar, preferably at 1 5mbar to 50mbar. The degree of concentration may be, for example, a concentration to 1/5, preferably to 1 /1 0 of the volume of the extraction solution obtained in step (a) or (b). The concentration in step (c) may also be, for example, up to dryness. Methods for concentrating extraction solutions are known to the skilled person and include evaporation on a rotary evaporator, thin film evaporator, vaccum centrifugal evaporator, vacuum band evaporator etc.. Fractionation step (d) preferably comprises filtration, centrifugation or chromatography, preferably chromatography. It will be evident to the skilled person that chromatography comprises techniques such as, e.g. column chromatography, planar chromatography, paper chromatography, thin layer chromatography, affinity chromatography, adsorption chromatography, liquid/liquid extraction etc.. In a preferred embodiment according to the invention, chromatography involves column chromatography, e.g. using polymeric resins, such as polystyrene, silica gels, aluminium oxides, dextrans etc., most preferably medium pressure liquid chromatography. A particularly preferred chromatography material is RP-C1 8 or RP-C8 silica (50 pm), e.g. obtainable from Macherey & Nagel. Preferably, chromatography is performed under reverse phase conditions, preferably using isocratic conditions for elution. Preferably, isocratic elution is effected using an alcohol/water solution, such as a methanol/water solution at a ratio of e.g. 80:20 (v/v), 70:30 (v/v) or 60:40 (v/v), preferably at a ratio of 70:30 (v/v).

The term "pooling" in the context of the present invention means "combining". Thus, in step (e) of the process according to the present invention, fractions containing lignan compounds are combined. Such fractions containing lignan compounds may be readily identified by the skilled person, for example, by applying diode array detection at 21 8 nm and/or HPLC methodology using, for example, a Phenomenex R Luna RP-C18 stationary phase column.

The optional concentration step (f) of the process for preparing a lignan composition according to the present invention may, for example, comprise freeze-drying, vacuum evaporation, vacuum band drying, spray drying etc. of the pooled lignan-containing fractions obtained in step (e). A particularly preferred concentration step comprises freeze drying. The degree of concentration in concentration step (f) is preferably at least 1/10, more preferably at least 1/20 of the volume of the pooled lignan fractions. The concentration may be, for example, up to dryness.

In a preferred embodiment, the process according to the present invention comprises the steps of:

(a) contacting lignan-containing plant material with an extraction solvent to obtain an extraction solution, wherein the extraction solvent is preferably a supercritical gas, such as supercritical C0₂, an organic solvent or an aqueous solution of an organic solvent, preferably alcohol or an aqueous solution of an alcohol, more preferably ethanol or an aqueous solution of ethanol, such as 90% ethanol aq. (v/v) or ethanol absolute,

(b) separating the plant material solids from said extraction solution, preferably comprising filtration or centrifugation, preferably filtration,

(c) optionally concentrating said extraction solution, preferably by evaporation or precipitation, preferably evaporation, e.g. to at least 1/5, preferably to at least 1/10 of the volume of the extraction solution obtained in step (b),

(d) fractionating the extraction solution, preferably by chromatography, more preferably by reversed phase chromatography, using a silica material,

(e) pooling lignan-containing fractions, and (f) optionally concentrating the pooled lignan-containing fractions

to obtain a lignan composition, wherein the lignan concentration in the lignan composition is at least 60% (w/w), preferably at least 65% (w/w), more preferably at least 70% (w/w), even more preferably at least 75% (w/w).

The lignan-containing plant material used in the process according to the present invention comprises at least one lignan compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, preferably comprises at least two lignan compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, more preferably comprises at least three lignan compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, even more preferably comprises at least four lignan compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, and most preferably comprises clusin, dihydrocubebin, hinokinin, yatein and cubebin.

The lignan-containing plant material used in the process according to the present invention is preferably derived from plant species selected from the group consisting of Piper cubeba, such as Piper cubeba L., Tarenna madagascariensis, Piper ribesioides, Piper c/usii, Juniperus phoenicea, such as Moroccan Juniperus phoenicea, Juniperus thurifera, such as Juniperus thurifera var. Africana, Chamaecyparis obtuse, and Lychnophora ericoides. Preferably, the plant material, which is derived from Piper cubeba, is Piper cubeba L. fruits, preferably unripe Piper cubeba L. fruits, most preferably Fructus Cubebae as defined in " Deutsches Arzneibuch 6" (DAB6). The term "plant material derived from plant species" preferably means that the plant material may be any part of the plant or a mixture of plant parts of the particular plant species it is derived from, such as the leaves, the flowers, the seeds, the fruits, unripe fruits, buds, stems, stalks, roots etc., or the entire plant. Particularly preferred plant parts may vary between the different applicable plants. For example, particularly preferred plant parts for Piper cubeba are the fruits, particularly preferred plant parts for Piper ribesioides are the fruits and/or stems, particularly preferred plant parts for Chamaecyparis obtusa are the leaves, particularly preferred plant parts for Lychnophora ericoides are the roots and/or stems. The skilled person is readily able to determine which plant parts are particularly useful for the process according the to the present invention, i.e. such plant parts comprising lignan compounds, by routine methods known in the art, such as by HPLC technology. Different plant species provide for different lignan compounds. Thus, Piper cubeba comprises clusin, dihydrocubebin, hinokinin, yatein and cubebin, Tarenna madagascariensis comprises hinokinin, cubebin and dihydrocubebin, Piper ribesioides comprises cubebin and hinokinin, Piper clusii comprises clusin, Juniperus phoenicea, such as Moroccan Juniperus phoenicea, comprises yatein, Juniperus thurifera, such as Juniperus thurifera var. Africana, comprises yatein, Chamaecyparis obtusa comprises yatein, and Lychnophora ericoides comprises hinokinin, cubebin and dihydrocubebin.

The plant material in the context of the present invention may be derived from one or more plant species. For example, the plant material may be derived from a mixture of plant species, preferably a mixture of plant species comprising clusin, dihydrocubebin, hinokinin, yatein and cubebin.

The process for preparing a lignan composition according to the present invention preferably results in a lignan composition comprising clusin, dihydrocubebin, hinokinin, yatein and cubebin as described herein. Thus, preferably, the plant material is selected such that all five lignan compounds are present in the plant material. For example, the plant material is preferably derived from:

Piper cubeba,

a mixture of Piper cubeba with any of the other plant species mentioned herein, such as a mixture of Piper cubeba and Piper clusii ox Tarenna madagascariensis etc.,

a mixture of Tarenna madagascariensis, Piper clusii and Juniperus phoenicea, such as Moroccan Juniperus phoenicea, Juniperus thurifera, such as Juniperus thurifera var. Africana, or Chamaecyparis obtuse,

a mixture of Lychnophora ericoides, Piper clusii and Juniperus phoenicea, such as Moroccan Juniperus phoenicea, Juniperus thurifera, such as Juniperus thurifera var. Africana, or Chamaecyparis obtuse, etc..

The process according to the present invention may comprise the step of adding lignan compound(s), e.g. as purified or synthesized lignan compounds, to the lignan composition, e.g. obtained in step (e) or (f). For example, in the event that the plant material does not comprise all of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin, the missing lignan may be added, e.g. as purified or synthesized lignan. In a particular preferred embodiment of the process according to the present invention, the lignan-containing plant material for preparing the lignan composition are fruits, preferably unripe fruits, from a Piper cubeba L. plant, and the process comprises the steps of:

(a) extracting Piper cubeba powder, e.g. C0₂-defatted Piper cubeba powder, with 90%- 100% (v/v) ethanol aq. in a drug solvent ratio of 1 :5 (w/w) by agitating the mixture for 1 - 4 hours, preferably 2 hours, at between about 1 5-30°C, preferably at between about 20- 25°C,

(b) separating plant solids by filtering the extract, preferably twice, preferably using a deep layer cellulose filter,

(c) concentrating the extract by evaporation, preferably on a rotary evaporator, e.g. to at least 1/10 of the volume of the extraction solution obtained from step (b),

(d) fractionating the extraction solution comprising the steps:

(d1 ) dissolving the concentrated extract in alcohol, preferably in ethanol, preferably in absolute ethanol, preferably in a ratio of 1 :4 (weight of extract : volume of solvent)

(d2) subjecting the solution obtained in step (d1 ) to medium pressure liquid chromatography using reverse phase conditions, e.g. using polymeric resins, such as polystyrene, silica gels, aluminium oxides, dextrans etc, preferably using silica material, such as RP-C18 silica (e.g. 50 μηι),

(d3) eluting under isocratic conditions, preferably using a methanol/water solution, preferably at a ratio of methanol : water of 70:30 (v/v),

(d4) collecting fractions of eluted material, preferably at a fraction size of between about 10-100 ml, preferably of about 50 ml,

(d5) detecting lignan-containing factions, preferably by applying diode array detection at 218 nm and/or HPLC technology,

(e) pooling lignan-containing factions, and

(f) concentrating the pooled lignan-containing fractions, preferably by evaporation, more preferably by freeze drying, preferably to at least 1/10 of the volume of the pooled lignan-containing fractions, most preferably to dryness

to obtain a lignan composition, wherein the lignan concentration in the lignan composition, preferably the lignan concentration of the lignans clusin, dihydrocubebin, hinokinin, yatein and cubebin together, is at least 60% (w/w), preferably at least 65% (w/w), more preferably at least 70% (w/w), even more preferably at least 75% (w/w). The lignan concentration may, for example, be determined using HPLC technology.

Preferably, the process according to the present invention further comprises the steps of: (g) adding one or more pharmaceutically acceptable excipients, diluents and/or carriers to the lignan composition obtained in step (e) or (0,

(h) optionally formulating the lignan composition obtained in step (f) or step (g) to obtain a pharmaceutical formulation, preferably a parenteral or non-parenteral formulation.

In a further aspect, the present invention provides a lignan composition obtainable by the process according to the present invention.

Thus, the present invention provides, for example, a lignan composition obtainable by a process comprising the steps of:

(b) separating the plant material solids from said extraction solution,

(c) optionally concentrating said extraction solution,

(d) fractionating the extraction solution obtained in step b) or c),

(e) pooling lignan containing fractions,

(f) optionally concentration the pooled lignan-containing fractions,

to obtain a lignan composition, wherein the lignan concentration in the lignan composition, preferably the lignan concentration of the lignans clusin, dihydrocubebin, hinokinin, yatein and cubebin together, is at least 60% (w/w), preferably at least 65% (w/w), more preferably at least 70% (w/w), even more preferably at least 75% (w/w),

(g) optionally adding one or more pharmaceutically acceptable excipients, diluents and/or carriers to the lignan composition obtained in step(e) or (0, and

(h) optionally formulating the lignan composition obtained in step (e), (f) or (g) to obtain a pharmaceutical formulation, preferably a parenteral or non-parenteral formulation. as described above.

In a preferred embodiment, the present invention provides a lignan composition obtainable by a process comprising the steps of:

(a) contacting lignan-containing plant material with an extraction solvent to obtain an extraction solution, wherein the extraction solvent is preferably a supercritical gas, such as supercritical C0₂, an organic solvent or an aqueous solution of an organic solvent, preferably alcohol or an aqueous solution of an alcohol, more preferably ethanol or an aqueous solution of ethanol, (b) separating the plant material solids from said extraction solution, preferably comprising filtration or centrifugation, preferably by filtration,

(c) optionally concentrating said extraction solution, preferably by evaporation or precipitation, preferably by evaporation, e.g. to at least 1/5, preferably to at least 1/10 of the volume of the extraction solution obtained in step (b),

(d) fractionating the extraction solution, preferably by chromatography, more preferably by reversed phase chromatography, e.g. using a silica material,

(e) pooling I ignan-containing fractions,

(f) optionally concentrating the pooled lignan-containing fractions,

(g) optionally adding one or more pharmaceutically acceptable excipients, diluents and/or carriers to the lignan composition obtained in step (e) or (f), and

(h) optionally formulating the lignan composition obtained in step (e), (f) or (g) to obtain a pharmaceutical formulation, preferably a parenteral or non-parenteral formulation as described above. In a particularly preferred embodiment, the present invention provides a lignan composition obtainable by a process comprising the steps of:

(a) extracting Piper cubeba powder, e.g. C0₂-defatted Piper cubeba powder, with 90%- 100% (v/v) ethanol aq. in a drug solvent ratio of 1 :5 (w/w) by agitating the mixture for 1 - 4 hours, preferably 2 hours, at between about 15-30°C, preferably at between about 20-

25°C,

(d) fractionating the extraction solution comprising the steps:

(d2) subjecting the solution obtained in step (d1 ) to medium pressure liquid chromatography using reverse phase conditions, e.g. using polymeric resins, such as polystyrene, silica gels, aluminium oxides, dextrans etc, preferably using silica material, such as RP-C1 8 silica (e.g. 50 pm),

(d5) detecting lignan-containing factions, preferably by applying diode array detection at 21 8 nm and/or HPLC technology,

(e) pooling lignan-containing factions, and

(h) optionally formulating the lignan composition obtained in step (f) or step (g) to obtain a pharmaceutical formulation, preferably a parenteral or non-parenteral formulation as described above.

Thus, the present invention provides, for example, a lignan composition obtainable by a process comprising or consisting of the method steps of Example 1 or 2 in combination with the method steps of Example 1 1 .

The present invention also provides the lignan composition obtainable by a process according to the present invention for use as a medicament, for example for use in the treatment or prevention of a tumour disease as defined herein, such as for use in the treatment or prevention of a hormone-dependent cancer, e.g. hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer.

Thus, for example, the present invention provides a lignan composition obtainable by the process according to the present invention for use in the treatment or prevention of prostate cancer, breast cancer, pancreatic cancer, preferably for use in the treatment of prostate cancer, such as hormone-dependent prostate cancer.

Furthermore, the present invention provides a pharmaceutical composition, a pharmaceutical formulation, and a medicament comprising the lignan composition according to the present invention and optionally one or more pharmaceutically acceptable excipients, carriers and/or diluents.

In a further aspect the present invention provides a composition for aiding in the prophylaxis or prevention of a tumor disease, preferably for aiding in the prophylaxis or prevention of cancer, more preferably for aiding in the prophylaxis or prevention of a hormone-dependent cancer, most preferably for aiding in the prophylaxis or prevention of prostate cancer.

The term "prophylaxis" as used within the contect of the compositions according to the present invention refers to the prevention of the occurrence of a tumour disease, such as cancer, e.g. hormone-dependent cancer types, e.g. androgen or estrogen-dependent cancer types, such as, e.g. prostate cancer.

The compositions according to the present invention comprise at least three, preferably four, most preferably five lignans selected from the group comprising clusin, yatein, dihydrocubebin, hinokinin and cubebein. Accordingly, the composition according to the present invention comprises at least clusin, dihydrocubebin and hinokinin, or at least dihydrocubebin, hinokinin and yatein, or at least hinokinin, yatein and cubein, or at least yatein, cubebin and clusin, or at least cubebin, clusin and dihydrocubebin, or at least cubebin, dihydrocubebin and hinokinin, or at least yatein, cubebin, dihydrocubebin, or preferably at least clusin, hinokinin, yatein and dihydrocubebin, or at least hinokinin, yatein, dihydrocubebin and cubebin, or at least clusin, yatein, dihydrocubebin and cubebin, or at least clusin, hinokinin, dihydrocubebin and cubein, or at least clusin, hinokinin, yatein and cubebin, most preferably clusin, hinokinin, yatein, dihydrocubebin and cubebin, wherein clusin is present in a concentration of at least 10% (w/w), preferably in a concentration of between 10% and 30% (w/w), and/or dihydrocubebin is present in a concentration of at least 5% (w/w), preferably in a concentration of between 5% and 20% (w/w), and/or hinokinin is present in a concentration of at least 15% (w/w), preferably in a concentration of between 15% and 50% (w/w), and/or yatein is present in a concentration of at least 2% (w/w), preferably in a concentration of between 2% and 15% (w/w), and/or cubebin is present in a concentration of at least 25% (w/w), preferably in a concentration of between 25% and 60% (w/w). Thus, in one embodiment, the present invention provides clusin for use in aiding in the prophylaxis or prevention of a tumour disease as disclosed above, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer, such as hormone-dependent prostate cancer.

Furthermore, the present invention provides dihydrocubebin for use in aiding in the prophylaxis or prevention of a tumour disease as disclosed above, preferably for aiding in the prophylaxis or prevention of cancer, more preferably for use in aiding in the prophylaxis or prevention of a hormone-dependent cancer, most preferably for use in aiding in the prophylaxis or prevention of prostate cancer, such as hormone-dependent prostate cancer.

Furthermore, the present invention provides hinokinin for use in aiding in the prophylaxis or prevention of a tumour disease as disclosed above, preferably for use in aiding in the prophylaxis or prevention of cancer, more preferably for use in aiding in the prophylaxis or prevention of a hormone-dependent cancer, most preferably for use in aiding in the prophylaxis or prevention of prostate cancer, such as hormone-dependent prostate cancer.

Furthermore, the present invention provides dihydrocubebin for use in aiding in the prophylaxis or prevention of a tumour disease as disclosed above, preferably for in aiding in the prophylaxis or prevention of cancer, more preferably for use in aiding in the prophylaxis or prevention of a hormone-dependent cancer, most preferably for use in aiding in the prophylaxis or prevention of prostate cancer, such as hormone-dependent prostate cancer. Furthermore, the present invention provides yatein for use in aiding in the prophylaxis or prevention of a tumour disease as disclosed above, preferably for aiding in the prophylaxis or prevention of cancer, more preferably for use in aiding in the prophylaxis or prevention of a hormone-dependent cancer, most preferably for use in aiding in the prophylaxis or prevention of prostate cancer, such as hormone-dependent prostate cancer.

Furthermore, the present invention provides cubebin for use in aiding in the prophylaxis or prevention of a tumour disease as disclosed above, preferably for aiding in the prophylaxis or prevention of cancer, more preferably for use in aiding in the prophylaxis or prevention of a hormone-dependent cancer, most preferably for use in aiding in the prophylaxis or prevention of prostate cancer, such as hormone-dependent prostate cancer. The compositions according to the present invention comprise at least three, preferably four, most preferably five lignans selected from the group comprising clusin, yatein, dihydrocubebin, hinokinin and cubebein as main constituents, i.e. the concentration of the lignan compounds together, preferably the concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together is at least 60% (w/w), preferably at least 65% (w/w), more preferably at least 70% (w/w) of the composition according to the invention.

In one embodiment, the compositions according to the present invention may be obtained by the inventive process for obtaining a lignan composition as disclosed above, e.g. the process for preparing a lignan composition according to the present invention, which preferably results in a lignan composition comprising at least clusin, dihydrocubebin, hinokinin, yatein and cubebin as described herein, e.g. the concentration of the lignan compounds is at least 60% (w/w), e.g. preferably at least 65% (w/w), e.g. more preferably 70% (w/w). The inventive compositions may be used e.g. in aiding in the prophylaxis or prevention of a tumour disease as disclosed above, e.g. preferably for use in aiding in the prophylaxis or prevention of cancer, e.g. more preferably for use in aiding in the prophylaxis or prevention of a hormone-dependent cancer, e.g. most preferably for use in aiding in the prophylaxis or prevention of prostate cancer, such as hormone-dependent prostate cancer.

Clusin, dihydrocubebin, hinokinin, yatein and cubebin may be present in the composition according to the present invention, e.g. in a concentration as disclosed above for the composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin. Thus, for example, clusin may be present in a concentration of at least 10% (w/w), e.g. in a concentration of between 10% and 30% (w/w), and/or dihydrocubebin may be present in a concentration of, e.g. at least 5% (w/w), such as in a concentration of between 5% and 20% (w/w), and/or hinokinin may be present in a concentration of, e.g. at least 1 5%, such as in a concentration of between 15% and 50% (w/w), and/or yatein may be present in a concentration of, e.g. at least 2% (w/w), such as in a concentration of between 2% and 15% (w/w), and/or cubebin may be present in a concentration of, e.g. at least 25%, such as in a concentration of between 25% and 60% (w/w), in the pharmaceutical composition according to the present invention.

In the composition according to the invention comprising at least three, or at least four, or at least five of the lignan compounds according to the present invention, the ratios of the lignans within the compositions may differ from the ratios of the Piper cubeba extracts according to the present invention, i.e. to the Piper cubeba fraction according to Example 1 1 (Pip.cub.40p2.001 .E03.S04). Accordingly, the compositions may comprise more or less of any one of the compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin than the Piper cubeba fraction according to the present invention, e.g. Pip.cub.40p2.001 .E03.S04. However, the ratios of the lignans in the pharmaceutical composition according to the invention may correspond to the ratios of the compounds in the inventive Piper cubeba fraction Pip.cub.40p2.001 .E03.S04.

The compositions according to the present invention may further comprise one or more pharmaceutically acceptable carriers. The term "pharmaceutically acceptable carrier" in the context of the present invention relates to one or more compatible solid or liquid fil lers or diluents, which are suitable for an administration to an animal, such as a mammal, e.g. a human. The term "carrier" relates to a natural or synthetic organic or inorganic component which is combined with a pharmaceutically active component in order to faci litate the application of the pharmaceutically active component. Preferably, carrier components are sterile liquids such as water or oils, including those which are derived from mineral oil, animals or plants, such as peanut oil, soy bean oil, sesame oil, sunflower oil, etc. Salt solutions, aqueous dextrose and glycerin solutions, as well as a Ringer's solution may also be used as aqueous carrier compounds. Particularly useful pharmaceutically acceptable carriers for use in the composition in the context of the present invention are emulsifying or solubilising substances, such as transcutol and/or Span20. Further useful pharmaceutically acceptable carriers are, for example, polyglycerins, polyoxyethylene glycols and their fatty acid esters like Macrogol glycerol hydroxystearate.

"Pharmaceutically acceptable diluents" as used in the context of the compositions of the present invention may, for example, be lactose, mannitol, sucrose, dextrose, cellulose, such as microcrystalline cellulose or powdered cellulose, starches, sorbitol, dibasic calcium phosphate, calcium carbonate and mixtures thereof.

The composositions according to the invention may also be used as a dietary or health supplement. Dietary or health supplement as used in the present invention refers to all forms of supplying an effective amount of the compositions according to the invention for consumption or administration. The compositions according to the invention, which are to be used as a dietary or health supplement may be provided, for example, as tablets, powders, chewable tablets, capsules, food additives, oral suspensions, enteral feeds, parenteral nutrition (for example, intravenous feeds), nutraceuticals and functional foods, preferably, the dietary or health supplement is in a form selected from but not limited to the group consisting of capsule, tablet and powder, e.g. for use in aiding in the prophylaxis or prevention of a tumour disease as disclosed above, e.g. preferably for use in aiding in the prophylaxis or prevention of cancer, e.g. more preferably for use in aiding in the prophylaxis or prevention of a hormone-dependent cancer, most preferably for use in aiding in the prophylaxis or prevention of prostate cancer, such as hormone-dependent prostate cancer.

Furthermore, the present invention provides a composition for use as a health or dietary supplement as disclosed above. Accordingly, the present invention provides for a composition comprising e.g. at least three, e.g. preferably at least four, e.g. most preferred at least five of the lignans clusin, yatein, hinokinin, dihydrocubebin and cubebin as disclosed above, or e.g. comprising a lignan composition obtainable by a process as disclosed above, or e.g. any one of the pharmaceutical compositions as disclosed above.

Furthermore the present invention provides for the use of a composition as health or dietary supplement as disclosed above. Accordingly, the present invention provides for the use of a composition comprising e.g. at least three, e.g. preferably at least four, e.g. most preferred at least five of the lignans clusin, yatein, hinokinin, dihydrocubebin and cubebin as disclosed above, or e.g. comprising a lignan composition obtainable by a process as disclosed above, or e.g. any one of the pharmaceutical compositions as disclosed above.

The present invention further provides for the use of a composition as disclosed above, in particular for the preparation of a nutritional product, which comprises the composition as disclosed obove, e.g. the lignan composition comprising at least yatein, clusin, dihydrocubebin, cubebin and hinokinin as disclosed above, or e.g. the composition which comprises the lignan composition obtainable by a process as disclosed above. Accordingly, any one of the pharmaceutical compositions as disclosed above may be used for the preparation of the nutritional product, or e.g. for use as a health or dietary supplement. The nutritional product according to the present invention may be e.g. foodstuffs, such as e.g. ganola bars, cookies, chewable tablets, capsules, food additives, such as powders, or gels, oral suspensions, enteral feeds, parenteral nutrition (for example, intravenous feeds), nutraceuticals, or beverage, such as e.g. milk, tea, coffee, emulsions, or liquid energy drinks, shakes, or smoothies.

Accordingly, the present invention provides a nutritional product comprising the composition as disclosed above, or any one of the pharmaceutical compositions as disclosed above, e.g. for use in aiding in the prophylaxis or prevention of a tumour disease as disclosed above, e.g. preferably for use in aiding in the prophylaxis or prevention of cancer, e.g. more preferably for use in aiding in the prophylaxis or prevention of a hormone-dependent cancer, e.g. most preferably for use in aiding in the prophylaxis or prevention of prostate cancer, such as hormone-dependent prostate cancer. In a further aspect, the present invention provides a method of treating or preventing a tumour disease as described above comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition according to the present invention, a therapeutically effective amount of a lignan composition according to the present invention, a therapeutically effective amount of a pharmaceutical composition, a pharmaceutical formulation or a medicament comprising the lignan composition according to the present invention, or a therapeutically effective amount of one or more compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, preferably selected from the group consisting of clusin, dihydrocubebin and hinokinin, more preferably selected from the group consisting of clusin and dihydrocubebin.

The tumour disease is preferably cancer, such as a hormone-dependent or hormone- independent cancer, preferably a hormone-dependent cancer, such as an androgen- or estrogen-dependent cancer as described herein. In a particularly preferred embodiment the method is for treating or preventing prostate cancer, such as hormone-dependent prostate cancer, breast cancer, such as hormone-dependent breast cancer, or pancreatic cancer.

Thus, the present invention provides, for example, a method of treating or preventing a tumour disease as described herein, preferably cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, more preferably prostate cancer, such as hormone-dependent prostate cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition comprising at least one, preferably at least two, such as two, three, or four compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin and optionally one or more pharmaceutically active excipients, carriers and/or diluents, e.g. in a combination and/or in concentrations as disclosed herein.

Furthermore, the present invention provides a method of treating or preventing a tumour disease as described herein, preferably cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, more preferably prostate cancer, such as hormone-dependent prostate cancer, comprising administering to a subject in need thereof a therapeutically effective amount of one or more, preferably of at least two, such as two, three, or four compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, preferably selected from the group consisting of clusin, dihydrocubebin and hinokinin, more preferably selected from the group consisting of clusin and dihydrocubebin, e.g. in any combination and/or in any concentration as defined herein. The compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin may be used alone or in combination. If used in combination, the compounds may be used together in a pharmaceutical composition as disclosed above or separately, for example, in separate pharmaceutical compositions, e.g. for concurrent or sequential administration and/or for the same or different administration routes.

Furthermore, the present invention provides a method of treating or preventing a tumour disease as described herein, preferably cancer, preferably selected from the group consisting of prostate cancer, breast cancer and pancreatic cancer, more preferably prostate cancer, such as hormone-dependent prostate cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a lignan composition obtainable by a process according to the present invention as described above, a pharmaceutical composition comprising a lignan composition according to the present invention, a pharmaceutical formulation comprising a lignan composition according to the present invention, or a medicament comprising a lignan composition according to the present invention.

In the context of the present invention, the term "subject" preferably relates to a "patient", an "individual", or an "animal", which terms preferably relate to a mammal. For example, mammals in the context of the present invention are humans, non-human primates, domesticated animals such as dogs, cats, sheep, cattle, goats, pigs, horses etc., laboratory animals such as mice, rats, rabbits, guinea pigs, etc. as well as animals in captivity such as animals of zoos. The term "animal" as used herein also includes humans. A preferred "subject" in the context of the present invention is a human.

The term "excipient" when used herein is intended to indicate all substances in a pharmaceutical formulation or pharmaceutical composition which are not pharmaceutically active ingredients such as, e.g. carriers, binders, lubricants, thickeners, disintegrants, surface active agents, preservatives, emulsifiers, buffers, flavouring agents, colorants, glidants, coatings or protective matrices etc. Binders may be, for example, acacia, alginic acid, carbomer, carboxymethyl cellulose sodium, ethyl cellulose, guar gum, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, hydroxyethyl cellulose, magnesium aluminium silicate, methyl cellulose, povidone, pregelatinized starch, sodium alginate, starch, dextrin, gelatine, hydrogenated vegetable oils, polymethacrylates, zein and the like. A non-limiting list of commonly used and accepted excipients is provided in "VOLUME 3B Guidelines Medicinal products for human use Safety, environment and information Excipients in the label and package leaflet of medicinal products for human use, July 2003" issued by the European Commission, which is hereby incorporated in its entirety into the disclosure of this invention.

The pharmaceutical compositions according to the present invention may comprise one or more pharmaceutically acceptable carriers. The term "pharmaceutically acceptable carrier" in the context of the present invention relates to one or more compatible solid or liquid fillers or diluents, which are suitable for an administration to an animal, such as a mammal, e.g. a human. The term "carrier" relates to a natural or synthetic organic or inorganic component which is combined with a pharmaceutically active component in order to facilitate the application of the pharmaceutically active component. Preferably, carrier components are sterile liquids such as water or oils, including those which are derived from mineral oil, animals or plants, such as peanut oil, soy bean oil, sesame oil, sunflower oil, etc. Salt solutions, aqueous dextrose and glycerin solutions, as well as a Ringer's solution may also be used as aqueous carrier compounds. Particularly useful pharmaceutically acceptable carriers in the context of the present invention are emulsifying or solubilising substances, such as transcutol and/or Span20. Further useful pharmaceutically acceptable carriers are, for example, polyglycerins, polyoxyethylene glycols and their fatty acid esters like Macrogol glycerol hydroxystearate.

"Pharmaceutically acceptable diluents" may, for example, be lactose, mannitol, sucrose, dextrose, cellulose, such as microcrystalline cellulose or powdered cellulose, starches, sorbitol, dibasic calcium phosphate, calcium carbonate and mixtures thereof.

The pharmaceutical compositions according to the present invention may, in some embodiments, comprise further pharmaceutically active agents, for example, a therapeutically active component such as an anti-tumour and/or anti-cancer agent. Examples for such optional further active agents are gemcitabine, irinotecan, 5-flurouracil, cisplatin etc. In addition or alternatively, the pharmaceutical composition may further comprise one or more agents which facilitate the uptake of the compounds used according to the present invention, e.g. which improve the bioavailability of the compounds, for example, if the compound or the composition is applied. Such agents are for example PGP inhibitors like e.g. Macrogol glycerol hydrostearate. A "pharmaceutically active agent" in the context of the present invention refers to a "pharmaceutically active ingredient", i.e. a compound which possesses therapeutic or preventive potential. For example, in the context of the present invention said therapeutic or preventive potential preferably relates to therapeutic or preventive activity against a tumour disease as described herein, e.g. against prostate cancer, such as against hormone-dependent prostate cancer. In the context of the present invention, clusin, dihydrocubebin, hinokinin, yatein and cubebin are pharmaceutically active agents.

The lignan compounds used in the present invention, the pharmaceutical composition according to the present invention, and the lignan composition according to the present invention may be formulated in any suitable way, e.g. depending on the intended administration route. For example, the lignan compounds used in the present invention, the pharmaceutical composition, and the lignan composition according to the present invention may be formulated as tablet, pill, capsule, suppository, powder, granules, solution, suspension etc. In one embodiment, the pharmaceutical composition is formulated in a solid formulation form, such as a tablet, pill, powder, granule, such as encapsulated granules etc. In another embodiment, the pharmaceutical composition is formulated in liquid form, such as in a solution or suspension. For preparing solid compositions such as tablets, the pharmaceutically active ingredient(s) is/are mixed with a pharmaceutical excipient or carrier, e.g. conventional tableting ingredients such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate or gums, and optionally pharmaceutical diluents to form a solid preformulation composition containing a homogenous mixture of the compound according to the present invention. The solid preformulation composition may then be subdivided into unit dosage forms. Tablets or pills may be coated or otherwise compounded to provide a dosage form affording the advantage of, e.g. prolonged action or improved delivery.

The liquid forms of the pharmaceutical compositions according to the present invention, e.g. for oral administration or administration by injection, include, for example, aqueous solutions, liposomal preparations, micro- or nano-emulsions, alcoholic solutions, optionally flavoured syrups, aqueous, alcoholic or oil suspensions, and optionally flavoured emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil etc.. The lignan compounds used in the present invention, the pharmaceutical compositions according to the present invention, or the lignan composition according to the present invention may be administered to an individual in need thereof, for example, a patient, by any suitable administration route, such as by oral, topical, rectal, vaginal, dermal, intra-tumoural, nasal, lingual, parenteral administration or administration by inhalation, insufflation, injection, infusion or by enema. Thus, the lignan compound used in the present invention, the pharmaceutical composition according to the present invention, or the lignan composition according to the present invention may be adapted, for example, for oral, topical, rectal, vaginal, dermal, intra-tumoural, nasal, lingual, parenteral administration or administration by inhalation, insufflation, injection, infusion or by enema. Furthermore, the method according to the present invention may comprise oral, topical, rectal, vaginal, dermal, intra-tumoural, nasal, lingual, parenteral administration or administration by inhalation, insufflation, injection, infusion or by enema. Preferred administration routes are oral and parenteral administration, such as intravenous, intramuscular, subcutaneous, intranodal, intralymphatic, intra-tumoural or intraperitoneal injection, most preferable is oral administration. The lignan compound used in the present invention, the pharmaceutical composition according to the present invention, or the lignan composition according to the present invention may also be directly applied to the tumour cells, e.g. the tumour tissue, for example, by injection into the tumour or cancer tissue. Thus, in a particularly preferred embodiment, the lignan compound used in the present invention, the pharmaceutical composition according to the present invention, or the lignan composition according to the present invention is adapted for parenteral administration, such as for intra-tumoural administration. Furthermore, preferably, the method according to the present invention comprises administering the lignan compound used in the present invention, the pharmaceutical composition according to the present invention, or the lignan composition according to the present invention by a parenteral route, e.g. by intravenous or intraperitoneal injection or by injection into the tumour or cancer tissue. In another embodiment, the method according to the present invention comprises oral administration of the lignan compound used in the present invention, of the pharmaceutical composition according to the present invention, or the lignan composition according to the present invention.

According to the present invention, the compounds and compositions according to the present invention are administered in a therapeutically effective amount. A "therapeutically effective amount" relates to an amount which - alone or in combination with further dosages - results in a desired reaction or a desired effect. In the case of the therapy of a particular disease or a particular condition, the desired reaction relates to the inhibition of the progress of the disease and/or the cure from the disease. This comprises the deceleration of the progress of the disease, in particular a disruption of the progression of the disease. An effective amount of the composition according to the present invention is dependent on the condition or disease, the severity of the disease, the individual parameters of the patient, including age, physiological condition, height and weight, the duration of the treatment, the type of an optionally accompanying therapy, the specific administration route, and similar factors. In case the reaction of a patient is insufficient with an initial dosage, higher dosages (or higher effective dosages which may be achieved by a more localized administration route) may be applied.

In particularly preferred embodiments of the present invention, the lignan compound, the pharmaceutical composition, or the lignan composition is administered repeatedly. Thus, the present invention provides the lignan compound used in the present invention, the pharmaceutical composition according to the present invention, or the lignan composition according to the present invention for use in the treatment or prevention of a tumour disease and the method of treating or preventing a tumour disease according to the present invention, wherein the compound or composition is administered repeatedly, for example, two times, three times, four times, five times, etc. For example, the compound or the compositions according to the present invention may be administered two times, three times, four times, five times, etc., preferably three times, at time intervals of between about 24 hours to 48 hours. The present invention is described in detail by the figures and examples below, which are used only for illustration purposes and are not meant to be limiting. Owing to the description and the examples, further embodiments which are likewise included in the invention are accessible to the skilled person.

EXAMPLES

Example 1 : Preparation of an ethanolic liquid extract Pip.cub.40p2.001.E03.F01 according to EP21 82966A1

1 758.3 g of CO2 defatted powder (< 2 mm) of Piper Cubeba was extracted in a stirred glass vessel with 8775.4 g ethanol absolute, corresponding to a drug solvent ration of 1/5 (w/w). The mixture was stirred on a magnetic stirrer at 500 rpm for 2 hours at room temperature before being filtered twice under vacuum using a Buechner funnel with a deep layer cellulose filter AF6 to give a clear brownish liquid extract. The extract Pip.cub.40p2.001 .E03.F01 was obtained in 8202.1 g with a solid content of 1 .89 % (DWC = 14.9 mg/mL) corresponding to a yield of native extract of 1 55 g (DER = 1 1 )

Example 2: Preparation of an ethanolic liquid extract Pip.cub.40p2.E02.F01 according to EP21 82966A1

1030 g of C02 defatted powder (< 2 mm) of Piper Cubeba was extracted in a stirred glass vessel with 51 50 g of 90 % ethanol (aq., w/w), corresponding to a drug solvent ration of 1 /5 (w/w). The mixture was stirred on a magnetic stirrer at 250 rpm for 2 hours at room temperature before being filtered twice times under vacuum using a Buechner funnel with a deep layer cellulose filter AF6 to give a clear brownish liquid extract. The extract Pip.cub.40p2.E02.F01 was obtained in 4778 g with a solid content of 1 .57 % (DWC = 12.8 mg mL) corresponding to a yield of native extract of 61 .3 g (DER = 1 7). Example 3: Preparation of a hydrophilic fraction Pip.cub.40p2.001 .E03.S01

A liquid extract, prepared as described in example 1 , was concentrated on a rotary evaporator at 40 °C, 70 mbar - 24 mbar to yield a brown, viscous oil. A portion of 5 g of this concentrated extract was dissolved in ethanol absolute to a total volume of ca. 20 mL and was then subjected to purification by medium pressure liquid chromatography using a hand packed glass column 36 x 460 mm filled with C8 reversed phase silica of 30 pm particle size (Macherey Nagel). Elution was achieved using a mixture of methanol/water 50:50 (v/v) as eluent (-500 ml), applying a flow rate of 40 ml / min and detecting with diode array at 21 8 nm. The collected hydrophilic fraction was concentrated on a rotary evaporator at 80 mbar and 40 °C and the remaining aqueous and cloudy solution was further dried on a freeze dryer affording 1 .01 g (20%) of the polar fraction Pip.cub.40p2.001 .E03.S01 as a highly viscous pale yellow oil. The total Lignan content of this material is < 5%.

Example 4: Preparation of a lipophilic fraction Pip.cub.40p2.001 .E03.S03

A liquid extract, prepared as described in example 1 , was concentrated on a rotary evaporator at 40 °C, 70 mbar - 24 mbar to yield a brown, viscous oil. A portion of 5 g of this concentrated extract was dissolved in ethanol absolute to a total volume of ca. 20 mL and was then subjected to purification by medium pressure liquid chromatography using a hand packed glass column 36 x 460 mm filled with C8 reversed phase silica of 30 pm particle size (Macherey Nagel). Elution was achieved using a mixture of methanol/water 50:50 (v/v) as initial eluent (-500 ml), applying a flow rate of 40 ml / min and detecting with diode array at 21 8 nm. Subsequently the mobile phase was changed to methanol/water 73:27 (v/v) and elution was continued for another ~1 .200 ml at which point the lignan fraction has been completely eluted. The solvent was then changed to ethanol absolute and elution was continued (-700 ml) thereby affording the unpolar extract constituents. This ethanolic fraction was evaporated to dryness on a rotary evaporator at 80 - 25 mbar and 40 °C giving 3.04 grams (60%) of Pip.cub.40p2.001 .E03.S03 as a brown oil. The total Lignan content of this material is < 5%.

Example 5: Preparation of Cubebin

500 g of C0₂ extracted powder from Piper cubeba fruits were mixed with 2.5 kg of ethanol absolute. The mixture was stirred for 2 hours at 40 °C and was then filtered using a deep layer cellulose filter AF15 from Filtrox followed by an AF40 filter. The filtrate was kept at 4 °C over night. The filtrate was concentrated on the rotary evaporator at 40 °C, 70 mbar - 24 mbar yielding 57.8 g of a brown oil. A preparative column 800G (92 x 378 mm) Silica HC, 50 pm was preconditioned with toluene/ethyl acetate 90:10 (v/v). 1 5 g of the sample was dissolved in 5 mL of mobile phase (total volume -20 ml) and was eluted using the equilibration condition at a flow rate of 80 ml/min applying diode array detection at = 280 nm and a fraction size of 50 ml during collection. This elution was continued until the first peak (hinokinin) was eluted and the signal was again lower than 7 % response. Then the mobile phase was changed to 70:30 (v/v) toluene/ethyl acetate and the following peak (yatein and cubebin) was collected. The relevant fractions of this section were analysed for cubebin content by HPLC methodology and were then pooled and dried on a rotary evaporator applying 40 °C and 1 mbar to give a yellow powder. Approximately 2 grams of this crude cubebin were heated to 65 °C in a water bath. Aceton/hexan 1 :1 was added slowly under reflux until the cubebin was dissolved (ca. 23 mL). Then the mixture was cooled down to 1 °C in an ice bath. The precipitate was filtered through a paper filter and washed with cold aceton/hexan 1 :1 until the precipitate a colourless crystalline solid. The residual solution was concentrated. The recrystallisation step was repeated twice. The obtained powders were kept in a desiccator over night to give 1 .85 gram of cubebin of a purity of 95.6 %, as determined by HPLC. The material was shown to be identical to an authentic reference standard of cubebin (de Pascoli et al., 2006, Phytochemistry 67: 735- 742).

Example 6: Preparation of Clusin 500 g of C0₂ extracted powder from Piper cubeba fruits were mixed with 2.5 kg of ethanol absolute. The mixture was stirred for 2 hours at 40 °C and was then filtered using a deep layer cellulose filter AF1 5 from Filtrox followed by an AF40 filter. The filtrate was kept at 4 °C over night. The filtrate was concentrated on the rotary evaporator at 40 °C, 70 mbar - 24 mbar yielding 57.8 g of a brown oil. A preparative column 800G (92 x 378 mm) Silica HC, 50 pm was preconditioned with toluene/ethyl acetate 90:10 (v/v). 15 g of the sample was dissolved in 5 mL of mobile phase (total volume -20 ml) and was eluted using the equilibration condition at a flow rate of 80 ml/min applying diode array detection at λ= 280 nm and a fraction size of 50 ml during collection. This elution was continued until the first peak (hinokinin) was eluted and the signal was again lower than 7 % response. Then the mobile phase was changed to 70:30 (v/v) toluene/ethyl acetate and the following peak (yatein and cubebin) was collected. Then the mobile phase was changed to 60:40 (v/v) toluene/ethyl acetate until the next peak (clusin and dihydrocubebin) was collected. The relevant fractions of this section were analysed for clusin content by HPLC methodology and were then pooled and dried on a rotary evaporator applying 40 °C and 1 mbar to give a yellow oil. 3.41 gram of this material was dissolved in 8 ml of dichloromethane and 1 ml (340 mg) of this solution was applied to a normal phase silica gel column (15 mm x 460 mm, 55 g of M60 Silica gel, 50 pm MN) and eluted using dichloromethane/methanol 97.5/2.5 (v/v) as eluent. Fractions were collected at 12 ml volume each and were analysed using HPTLC methodology (dichloromethane/methanol 95/5, detection reagent: sulphuric acid). The relevant fractions were pooled and concentrated on the rotary evaporator giving 730 mg of clusin of a purity 94.7 % (combined yield after processing the entire 3.41 g starting material).

The analytical data of the material were in full agreement with literature reported data for clusin (Koul et al., 1983 Phytochemistry 22: 999-1000).

Example 7: Preparation of Dihydrocubebin

500 g of C0₂ extracted powder from Piper cubeba fruits were mixed with 2.5 kg of ethanol absolute. The mixture was stirred for 2 hours at 40 °C and was then filtered using a deep layer cellulose filter AF1 5 from Filtrox followed by an AF40 filter. The filtrate was kept at 4 °C over night. The filtrate was concentrated on the rotary evaporator at 40 °C, 70 mbar - 24 mbar yielding 57.8 g of a brown oil. A preparative column 800G (92 x 378 mm) Silica HC, 50 pm was preconditioned with toluene/ethyl acetate 90:10 (v/v). 15 g of the sample was dissolved in 5 mL of mobile phase (total volume -20 ml) and was eluted using the equilibration condition at a flow rate of 80 ml/min applying diode array detection at = 280 nm and a fraction size of 50 ml during collection. This elution was continued until the first peak (hinokinin) was eluted and the signal was again lower than 7 % response. Then the mobile phase was changed to 70:30 (v/v) toluene/ethyl acetate and the following peak (yatein and cubebin) was collected. Then the mobile phase was changed to 60:40 (v/v) toluene/ethyl acetate until the next peak (clusin and dihydrocubebin) was collected. The relevant fractions of this section were analysed for dihydrocubebin content by HPLC methodology and were then pooled and dried on a rotary evaporator applying 40 °C and 1 mbar to give a yellow oil. 3.41 gram of this material was dissolved in 8 ml of dichloromethane and 1 ml (340 mg) of this solution was applied to a normal phase silica gel column (15 mm x 460 mm, 55 g of M60 Silica gel, 50 pm MN) and eluted using dichloromethane/methanol 97.5/2.5 (v/v) as eluent. Fractions were collected at 12 ml volume each and were analysed using HPTLC methodology (dichloromethane/methanol 95/5, detection reagent: sulphuric acid). The relevant fractions were pooled and concentrated on the rotary evaporator giving 410 mg of dihydrocubebin of a purity >98 % (combined yield after processing the entire 3.41 g starting material).

The analytical data of the material were in full agreement with literature reported data for Dihydrocubebin (Koul et al., 1983, Phytochemistry 22, 999-1000).

Example 8: Preparation of Hinokinin

500 g of C0₂ extracted powder from Piper cubeba fruits were mixed with 2.5 kg of ethanol absolute. The mixture was stirred for 2 hours at 40 °C and was then filtered using a deep layer cellulose filter AF15 from Filtrox followed by an AF40 filter. The filtrate was kept at 4 °C over night. The filtrate was concentrated on the rotary evaporator at 40 °C, 70 mbar - 24 mbar yielding 57.8 g of a brown oil. A preparative column 800G (92 x 378 mm) Silica HC, 50 pm was preconditioned with toluene/ethyl acetate 90:10 (v/v). 15 g of the sample was dissolved in 5 mL of mobile phase (total volume -20 ml) and was eluted using the equilibration condition at a flow rate of 80 ml/min applying diode array detection at = 280 nm and a fraction size of 50 ml during collection. This elution was continued until the first peak (hinokinin) was eluted and the signal was again lower than 7 % response. The relevant fractions of this section were analysed for hinokinin content by HPLC methodology and were then pooled and dried on a rotary evaporator applying 40 °C and 1 mbar to give a yellow oil.

4.70 gram of this material was dissolved in 20 ml of ethanol this solution was applied to a reversed phase silica gel column (49 mm x 460 mm, 500 g of RP-C18 silica, 50 pm MN) and eluted at 40 ml /min using a step gradient of 40/60 water/methanol (v/v) and 35/65 water/methanol (v/v) applying diode array detection at 280 nm and a fraction size of 50 ml during collection. Fractions were analysed using HPLC methodology and the relevant fractions were pooled and concentrated on the rotary evaporator (40°C, 70 mbar) to give a milky aqueous solution which was further freeze dried to afford the target compound in a yield of 600 mg and a purity of >98 %. The material was shown to be identical to an authentic reference standard of Hinokinin (Lopes et al., 1983, Phytochemistry 22: 1 51 6-1 518).

Example 9: Preparation of Yatein 500 g of C0₂ extracted powder from Piper cubeba fruits were mixed with 2.5 kg of EtOH. The mixture was stirred during 2 hours at 40 °C and then filtered on a deep layer cellulose filter AF1 5 from Filtrox followed by a AF40 filter. The filtrate was kept at 4 °C over night. The filtrate was concentrated on the rotary evaporator at 40 °C, 70 mbar - 24 mbar. Yielding 57.8 g of a brown oil. A preparative column 800G (92 x 378 mm) Silica HC, 50 pm was preconditioned with toluene/ethyl acetate 90:10 (V/V). The fraction size was 50 ml. The sample 15 g dissolved with 5 mL of mobile phase (V~20 ml) was applied. Flow rate 80 ml/min., λ = 280 nm until the first peak (hinokinin) was eluted and the signal lower than 7 %. Then the mobile phase was changed to toluene/ethyl acetate 70:30 (v/v) and the second peak (yatein and cubebin) was collected. The relevant fractions of this section were analysed for yatein content by HPLC methodology and were then pooled and dried on a rotary evaporator applying 40 °C and 1 mbar to give a yellow oil.

1 .07 g of this material was dissolved in 10 ml of methanol and 3 ml (325 mg) of this solution was applied to a reversed phase silica gel column (1 5 mm x 460 mm, 50 g of RP-C1 8 silica gel, 50 pm MN) and eluted using gradient from 90/100 water/methanol (v/v) to 40/60 water/methanol (v/v) applying diode array detection at 218 nm and a fraction size of 12 ml during collection. The relevant fractions were pooled and concentrated on the rotary evaporator giving 120 mg of yatein of a purity of 96 % (combined yield after processing the entire 1 .07 g starting material).

The analytical data of the material were in full agreement with literature reported data for yatein (Tanoguchi et al., 1987, Chem. Pharm. Bull. 35: 41 62-4168).

Example 10: Preparation of a lignan fraction Pip.cub.40p2.E07.S02 A liquid extract, prepared as described in example 1 , was concentrated on a rotary evaporator at 40 °C, 70 mbar - 24 mbar to yield a brown, viscous oil. A portion of 5 g of this concentrated extract was dissolved in ethanol absolute to a total volume of ca. 20 mL and was then subjected to purification by medium pressure liquid chromatography using a hand packed glass column 36 x 460 mm filled with C8 reversed phase silica of 30 pm particle size (Macherey Nagel). Elution was achieved using a mixture of methanol/water 50:50 (v/v) as initial eluent (-500 ml), applying a flow rate of 40 ml / min and detecting with diode array at 218 nm. Subsequently the mobile phase was changed to methanol/water 70:30 (v/v) elution was continued for another -1 .800 ml at which point the lignans have been completely eluted. Suitable, lignan containing, fractions of the collected eluent were concentrated on a rotary evaporator at 80 mbar and 40 °C. The remaining aqueous and cloudy solution was further dried on a freeze dryer affording 1 .27 g (25%) of the Lignan fraction Pip.cub.40p2.E07.S02 as a highly viscous pale yellow oil. The total Lignan content of this material was determined as >70%.

Example 1 1 : Preparation of a lignan fraction Pip.cub.40p2.001 .E03.S04

A liquid extract, prepared as described in example 1 , was concentrated on a rotary evaporator at 40 °C, 70 mbar - 24 mbar to yield a brown, viscous oil. A portion of 5 g of this concentrated extract was dissolved in ethanol absolute to a total volume of ca. 20 mL and was then subjected to purification by medium pressure liquid chromatography using a reversed phase conditions (49 mm x 460 mm, 500 g of RP-C18 silica, 50 pm MN) and eluted at 40 ml /min using isocratic conditions of methanol/water 70:30 (v/v) applying diode array detection at 218 nm and a fraction size of 50 ml during collection. The run was repeated using identical conditions for a total of 1 runs corresponding to a total of 55 g processed native extract. Fractions were analysed using HPLC methodology and the relevant fractions were pooled and concentrated on the rotary evaporator (40°C, 70 mbar) to give a milky aqueous solution. The remaining aqueous and cloudy solution was further dried on a freeze dryer affording 13.09 g (24%) of the Lignan fraction Pip.cub.40p2.001 .E03.S04 as a highly viscous pale yellow oil. The total Lignan content of this material was determined as >70%.

Example 12: Preparation of a lignan fraction Pip. cub.05.1 .S1

This Lignan fraction was obtained by supercritical extraction of Piper cubeba fruits with carbon dioxide recycle and two step separation. Prior to processing, the dried raw material was carefully ground (and heat generation avoided) and placed into a high pressure extractor. The system was subsequently pressurized and C02 passed through the material at 300 bar and 85°C. The extracts were collected by two-step partial depressurisation in two separator (separator 1 with waxy/actives, separator 2 with volatile oil). From 431 g of powdered fruits 41 g of volatile oils have been removed and 39 g of the Lignan Fraction Pip. cub.05.1 .S1 has been obtained while resulting in a remaining 329 g of residue. Analysis showed a Lignan content of 24.4% in Pip. cub.05.1 .S1 and -90% of the absolute Lignan content of the fruits has been extracted into the target extract Pip.cub.05.1 .S1 .

Example 1 3: Preparation of a lignan fraction Pip. cub.05.2. S1

This Lignan fraction was obtained by supercritical extraction of Piper cubeba fruits with carbon dioxide recycle and two step separation. Prior to processing, the dried raw material was carefully ground (and heat generation avoided) and placed into a high pressure extractor. The system was subsequently pressurized and C02 passed through the material at 300 bar and 90°C. The extracts were collected by two-step partial depressurisation in two separator (separator 1 with waxy/actives, separator 2 with volati le oil). From 410 g of powdered fruits 46 g of volatile oils have been removed and 38 g of the Lignan Fraction Pip. cub.05.2. SI has been obtained while resulting in a remaining 314 g of residue. Analysis showed a Lignan content of 28.3% in Pip. cub.05.2. S1 and -90% of the absolute Lignan content of the fruits has been extracted into the target extract Pip. cub.05.2. S1 .

Example 14: Preparation of a lignan fraction Pip. cub.05.3. S1

This Lignan fraction was obtained by supercritical extraction of Piper cubeba fruits with carbon dioxide recycle and two step separation. Prior to processing, the dried raw material was carefully ground (and heat generation avoided) and placed into a high pressure extractor. The system was subsequently pressurized and C02 passed through the material at 800 bar and 85 °C. The extracts were collected by two-step partial depressurisation in two separator (separator 1 with waxy/actives, separator 2 with volatile oil). From 434 g of powdered fruits 52 g of volatile oils have been removed and 46 g of the Lignan Fraction Pip. cub.05.3. SI has been obtained while resulting in a remaining 329 g of residue. Analysis showed a Lignan content of 27.7% in Pip. cub.05.3. S1 and -90% of the absolute Lignan content of the fruits has been extracted into the target extract Pip. cub.05.3. S1 . Example 15: Preparation of a lignan fraction Pip. cub.05.4. S1

This Lignan fraction was obtained by supercritical extraction of Piper cubeba fruits with carbon dioxide recycle and two step separation. Prior to processing, the dried raw material was carefully ground (and heat generation avoided) and placed into a high pressure extractor. The system was subsequently pressurized and C02 passed through the material at 800 bar and 85°C. The extracts were collected by two-step partial depressurisation in two separator (separator 1 with waxy/actives, separator 2 with volatile oil). From 41 Og of powdered fruits 64g of volatile oils have been removed and 28 g of the Lignan Fraction Pip. cub.05.4. S1 has been obtained while resulting in a remaining 304g of residue. Analysis showed a Lignan content of 44.0% in Pip. cub.05.4. S1 and -90% of the absolute Lignan content of the fruits has been extracted into the target extract Pip.cub.05.4.S1 .

Example 16: Preparation of a pre-formulation Pip.cub.40p2.001 .E03.C03-01

To 2304.7g of the fluid piper extract (prepared as in example 1 and containing 43.56g native extract) were added 87.06g Span 20 and 8.72 g Transcutol HP corresponding to a ratio of 1/2/0.2. The ethanol was evaporated under reduced pressure (80 to 25 mbar) and a bath temperature of 40°C. 1 56.22g of a brownish encapsulatable mass was obtained.

The 51 .57g native extract obtained was adjusted with 1 6.08g Span 20 and 1 .62g Transcutol HP in respect of the 1/2/0.2 ratio. The mixture was mixed under reduced pressure (25 mbar) and a bath temperature of 40°C for 90 min. 1 63.78 g of a brownish encapsulatable mass was obtained. The content of native extract was found to be 30.7% (w/w).

PHARMACOLOGICAL EXAMPLES IN CELL CULTURE

Example 1 7: Effect of the individual lignan on LNCaP cellular growth The human prostate cancer cell line LNCaP was cultivated in culture medium containing 1 0% FBS and kept in a humidified incubator at 37°C and 5% C0₂. Cultures used in subsequent experiments were passaged less than 20 times. The cells were seeded at 10,000 cells in 96- well plate and left to adhere overnight. The individual lignans, prepared according to examples 5 to 9 respectively, were serially diluted in ethanol to the corresponding concentrations before being added to the cells for a total of 48 hours. The cells were pulsed with a tracer, bromoxyuridine, BrdU 18 hours before the end of the treatment. The plates were then processed with the Cell Proliferation ELISA, BrdU (colorimetric) assay kit (Roche; Mannheim, Germany) according to manufacturer's instructions. Actively proliferating cells incorporated the BrdU could then be detected. The anti-proliferation effect observed on the treated cells was taken as a percentage to the solvent control treated cells.

Results

According to Fig.1 , the results indicate that all the lignans were able to inhibit cell proliferation of the human prostate cancer cell line LNCaP at relatively low concentrations. The most potent lignan in reducing cellular growth of the LNCaP cell line was dihydrocubebin, followed by clusin, yatein, hinokinin and cubebin.

Example 1 8: Effects of lignans versus the lignan fraction on LNCaP and HPAFII cellular growth

The anti-proliferative assay was performed similar to the above example 1 7. The lignan fraction (Pip cub.40p2.001 .E03.S04) and lignan compounds prepared according to examples 5,6,7,8,9 and 1 1 were tested on the LNCap cell line and the human pancreatic cell line HPAFII.

Results

The lignan fraction appeared to inhibit LNCaP (Fig.2) and HPAFII (Fig.7) cellular growth similar or more effective than the individual lignans at the same dose of 10 g/ml. The IC₅₀ values derived from the LNCaP and HPAFII anti-proliferative studies demonstrated that the single lignan compounds were inferior compared to the lignan fraction, with the exception of dihydrocubebin, which appeared to result in a greater inhibition of LNCaP cell growth than the lignan fraction. It must be noted that the corresponding concentration of the lignans in the lignan fraction were considerably lower than the IC₅₀ values of the individual lignans. (E.g. There is 0.49 g/ml of dihydrocubebin in 5.227 g/ml of lignan fraction). This indicates that there are synergistic effects of the 5 lignans.

Example 1 9: Lignan composition in various lignan fraction preparations

Several fractionation techniques were explored in order to determine the possible ranges of the lignans within the lignan fraction composition. The samples were prepared according to examples 10-14, the sample composition is shown in the table below. The spiked fractions were prepared by spiking with the corresponding percentage of the lignan for every one of them (e.g. the cubebin content in 100pg/ml Pip.cub.40p2.001 .E03.S04 is 38.9pg/ml, the spiked fraction comprises 77.8pg/ml of cubebin in 1 38.9pg/ml, resulting in the cubebin percentage of 56% in the spiked solution).

Example 20: Additive method of testing potential additive effect of combinations of lignans on LNCaP's cellular growth

The anti-proliferative assay was performed similar to the above example 1 7. The lignan fraction (Pip.cub.40p2.001 .E03.S04) and lignans prepared according to examples 5 to 1 1 were tested.

Results

The addition of the individual lignans did not dramatically increase the potency of the lignan fraction indicating that the efficacy of the lignan fraction is not attributed to a certain concentration ratio of the 5 lignans in the lignan fraction. The variations in the relative content of individual lignans in the lignan fraction did not significantly affect the anti-proliferative effect of the lignan fraction. Example 21 : Effects of 3 different fractions versus the plant extract on cancer cel ls' growth)

The anti-prol iferative assay was performed simi lar to the above example 1 7. The samples were prepared accordi ng to examples 1 , 3, 4 and 1 1 were tested.

Results

The extract was fractionated into three main components; hydrophi lic, lignan and l ipophi l ic fractions. As the data suggested, the lignan fraction appeared to retain most of the bioactivity whereas the other two fractions contribute minimally to the anti-proliferative effects. This clearly indicates that the lignans are the mai n active moieties in the extract. The potent effects of the l ignan fraction were more significant in other cancer cel l types (e.g. breast and pancreatic cancer) when compared to the extract.

Example 22 : Effects of l ignan fraction versus the plant extract on cancer cells colony formation The samples prepared according to examples 2 and 1 1 were tested agai nst cel l suspensions of 5 human tumour xenografts of prostate cancer i.e. 22 Rv1 , DU-1 45, MRI-1 579, PC-3 and PC-3M to investigate their potential to i nhibit ex vivo colony formation of cel ls with the abi lity to grow anchorage independently in semi-solid medium. The clonogenic assay was performed in a 24-well format according to a modified two-layer soft agar assay introduced by Hamburger & Salmon (Hamburger & Salomon, Science 197, 461 -463 (1977)). Each test well contained three layers of equal volume: 2 layers of semi-solid medium (bottom and top layer), and one layer of medium supernatant, with or without test compound. The bottom layer consisted of 0.2 ml/well IMDM (supplemented with 20% (v/v) fetal calf serum, 0.01 % (w/v) gentamicin) and 0.75% (w/v) agar. 8,000 to 20,000 cells were added to 0.2 ml of the same culture medium supplemented with 0.4% (w/v) agar and plated in 24-well plates onto the bottom layer. The test compounds were applied by continuous exposure (drug overlay) in 0.2 ml culture medium. The drug overlay was added to the culture as 3-fold concentrated solution repeatedly, i.e. 24 hours after seeding the cells for first time, and again on day 3 by replacement of the culture supernatant. Every plate included six untreated control wells and drug-treated groups in triplicate at 6 concentrations. Cultures were incubated at 37°C and 7.5% C02 in a humidified atmosphere for up to 20 days and monitored closely for colony growth using an inverted microscope. Within this period, ex vivo tumour growth led to the formation of colonies with a diameter of > 50 pm. At the time of maximum colony formation (based on internal historical data, range 4 - 21 days), counts were performed with an automatic image analysis system (BIOREADER 5000-W , Biosys GmbH). 24 hours prior to evaluation, vital colonies were stained with a sterile aqueous solution of 2-(4-iodophenyl)-3-(4- nitrophenyl)-5-phenyltetrazolium chloride (1 mg/ml, 100 μΙ/well) (Alley MC et al.. Life Sci. 31 , 3071 -3078 (1982)).

Results

Both PIPER extract (Example 2) and lignan fraction (Example 1 1 ) were characterized for their ability to inhibit anchorage-independent growth and in vitro colony formation of tumour cells in semi-solid medium. The fraction inhibited colony formation of several tumour models in a concentration-dependent manner whereas the Piper extract only inhibited the PC-3M cell line. Thus, lignan fraction was identified to be more potent than the original extract PC40p2E2F1 (described in EP2182966A1 ) and the active principle, which shows anti-tumour activity, seems to be enriched. Example 23: The pharmacokinetics of the lignan fraction versus hinokinin, a single lignan

The purpose of this study was to determine the pharmacokinetic behaviour of the lignan fraction and hinokinin in the plasma over time after oral administration in mice. Hinokinin was also applied intravenously as a control.

Male SCID-beige mice bearing the LNCaP tumours were randomized into 3 groups. Group 1 received Hinokinin (2.5mg/kg) administrated intravenously, Group 2 received Hinokinin 35.5 mg/kg orally and Group 3 was treated orally with the lignan fraction, Pip.cub.40p2.001 .E03.C03; prepared according to example 1 1 (156.7mg/kg). Group 2 and 3 were dosed with identical amounts of hinokinin. Hinokinin and the lignan fraction were dissolved in vehicle (Span20 Transcutol HP mix) before being orally given to Group 2 and 3, respectively. In order to monitor the plasma concentrations of hinokinin overtime, animals were sacrificed after 5,10,30,60,120 and 240 minutes. All plasma samples (calibration standards, QCs and unknown samples) were supplemented with 100 μΙ of internal standard solution (Diazepam: 1 50 ng/ml in acetonitrile) and shaken again for 10 seconds. After a centrifugation step (10 minutes at 5600g, 20°C) an aliquot of the supernatant was diluted with 1 volume of water, transferred to 200 μΙ sampler vials and subsequently subjected to LC-MS analysis

After the iv application of 2.5 mg/kg Hinokinin, the initial concentration was extrapolated to 141 1 ng/ml in plasma. The terminal elimination half life was calculated to 1 .3 hours. The oral application of 35.5 mg/kg Hinokinin led to maximal concentrations in plasma of 1982 ng/ml 2 hours post dose. The half -life, T1/2, was estimated to 2.2 hours and, whether area under curve (AUC) (0-tz) or AUC(O-oo) was considered, the bioavailability calculated to 108 and 1 70%, respectively The oral application of a lignan fraction corresponding to a dose of 35.5 mg/kg active substance was followed by a Cmax of 9834 ng/ml, reached 2 hours post dose. T1/2 was estimated to 1.8 hours and, whether AUC(O-tz) or AUC(O-oo) was considered, the bioavailability was calculated to 456 and 676%, respectively. The results as shown in Fig. 5 indicate that the oral administration of the lignan fraction resulted in substantially elevated levels of Hinokinin in the plasma as compared to oral intake of Hinokinin alone. This suggests a better bio-availability of Hinokinin (and any of the other 4 lignans individually) when administering the lignan in the form of a lignan fraction of at least two different lignans from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, if compared to administering a single lignan.

Example 24: Tissue distribution of lignans The purpose of this experiment was to determine the levels of lignans in various target organs when mice were orally treated with the extract (450mg/kg) versus a lignan content equivalence lignan fraction dose (157mg/kg); Pip.cub.40p2.001 .E03.C03; prepared according to example 1 6. Male SCID-Beige mice were treated with the test samples at the above mentioned dosages 3 times daily for 5 days before sacrifice. The prostates, adrenal glands and brains were harvested, processed and subsequently subjected to LC-MS analysis. The results are shown in Fig. 6.

Despite the reduced administered dose, the lignan fraction was able to deliver similar if not higher levels of the lignans especially hinokinin and yatein compared to the extract, PIPER VKM (Example 16). It appeared that the lignans were selectively accumulated in the prostate and adrenal glands, which are the target organs for the treatment of prostate cancer.

Example 25: Anti-proliferative effect of different lignan combinations on cell lines.

The human prostate cancer cell line LNCaP and PC-3M were cultivated in cell culture medium containing 10% FBS and kept in a humidified incubator at 37°C and 5% C02. Cells were seeded in 96-well plates (10000 LNCaP cells/well, 5000 PC-3M cells/well) and incubated for 24h prior to the addition of lignan combinations. Different lignan combinations as indicated in Fig. 8 (A), (B) were serially diluted in ethanol to the corresponding concentration. For experiments in which the lignan combinations were used (as indicated) the concentrations of the single compounds was adapted to 5pg ml, 3.3pg/ml and 2.5pg/ml if 2, 3 or 4 lignans were used, such that a final concentration of 10μg/ml total lignans per well was reached. The diluted samples were then added to the cells and incubated for 48h. The cells were then pulsed with bromoxyuridine and incubated for additional 18 hours. For the quantification of proliferating cells the amount of cells incorporating the BrdU was measured with the Cell proliferation ELISA (BrdU, colorimetric) assay kit (Roche) according to the manufacturers instructions. Proliferation is expressed as percentage of the solvent control treated cells (solvent control treated cells =100%). Results:

According to Fig.8 (A), (B) all combinations of lignans were able to decrease the proliferation of LNCaP cells. Combinations of three, four lignans were most effective. The extent of inhibition was similar to the lignan fraction Pip.cub.40p2.001 .E03.S04 comprising clusin, yatein, cubebin, dihydrocubebin and hinokinin as main constituents. In the PC-3M cell line the combination of three or four lignans was most effective and comparable to the lignan fraction, but the effect was not as pronounced as in the LNCaP cells.

On comparison of the relative extent of the growth inhibition at the given concentrations of 10ug/ml of total lignans it becomes evident that the incubation with three to four lignans results in overall better effects than incubation with a combination of two lignans in the prostate cancer cell lines LNCaP and PC-3M. The effects were similar with respect to the inhibition found with the lignan fraction Pip.cub.40p2.001 .E03.S04. The result indicates a synergistic effect on cell proliferation as the total amount of lignans was identical in all experiments. The experiments also show that the hormone-dependent prostate cancer cell line LNCaP was more sensitive to lignans than the cell line PC3-M, a sub line derived from the hormone refractory PC-3 prostate cancer cell line.

Claims

1 . Pharmaceutical composition comprising at least one compound selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, and optionally one or more pharmaceutically acceptable excipients, carriers, and/or diluents.

2. The pharmaceutical composition according to claim 1 , wherein the at least one compound is selected from the group consisting of clusin, dihydrocubebin and hinokinin.

3. The pharmaceutical composition according to claim 1 or 2, wherein the at least one compound is selected from the group consisting of clusin and dihydrocubebin.

4. The pharmaceutical composition according to any one of claims 1 -3 for use as a medicament.

5. The pharmaceutical composition according to any one of claims 1 -4 for use in the treatment or prevention of a tumour disease.

6. The pharmaceutical composition according to claim 5, wherein the tumour disease is cancer.

7. The pharmaceutical composition according to claim 5 or 6, wherein the tumour disease or cancer is hormone-dependent, preferably estrogen- or androgen-dependent.

8. The pharmaceutical composition according to any one of claims 1 -7 for use in the treatment or prevention of prostate cancer, breast cancer or pancreatic cancer, preferably for use in the treatment of prostate cancer, most preferably for the treatment or prevention of hormone-dependent prostate cancer.

9. The pharmaceutical composition according to any one of claims 1 -8 comprising at least two compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin

10. The pharmaceutical composition according to any one of claims 1 -9 comprising yatein and clusin and at least one further compound selected from the group consisting of dihydrocubebin, hinokinin and cubebin.

1 1 . The pharmaceutical composition according to any one of claims 1 -1 Ocomprising at least three compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin.

12. The pharmaceutical composition according to any one of claims 1 -1 1 comprising at least three compounds selected from the group consisting of clusin, dihydrocubebin, yatein and cubebin.

13. The pharmaceutical composition according to any one of claims 1 -12 comprising three compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, optionally comprising one or two further lignans from said group, such that the composition comprises at least three, optionally 4 or all of the lignans from said group.

14. The pharmaceutical composition according to any one of claims 1 - 1 3, wherein the composition comprises clusin, yatein and dihydrocubebin.

15. The pharmaceutical composition according to any one of claims 1 -12 comprising at least four compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin.

1 6. The pharmaceutical composition according to any one of claims 1 -1 5 comprising at least two, but not more than four compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin.

1 7. The pharmaceutical composition according to any one of claims 1 -1 5 comprising clusin, dihydrocubebin, hinokinin, yatein and cubebin.

18. The pharmaceutical composition according to any one of claims 1 -1 7, wherein clusin is present in a concentration of at least 10% (w/w), preferably in a concentration of between 10% and 30% (w/w).

19. The pharmaceutical composition according to any one of claims 1 -18, wherein dihydrocubebin is present in a concentration of at least 5% (w/w), preferably in a concentration of between 5% and 20% (w/w).

20. The pharmaceutical composition according to any one of claims 1 -1 9, wherein hinokinin is present in a concentration of at least 1 5% (w/w), preferably in a concentration of between 15% and 50% (w/w).

21 . The pharmaceutical composition according to any one of claims 1 -20, wherein yatein is present in a concentration of at least 2% (w/w), preferably in a concentration of between 2% and 15% (w/w).

22. The pharmaceutical composition according to any one of claims 1 -21 , wherein cubebin is present in a concentration of at least 25% (w/w), preferably in a concentration of between 25% and 60% (w/w).

23. The pharmaceutical composition according to any one of claims 1 -22, wherein the concentration of lignan compounds together, preferably the concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together is at least 60% (w/w), preferably at least 65% (w/w), more preferably at least 70% (w/w).

24. The pharmaceutical composition according to any one of claims 1 -23, which is essentially free from proteins and/or amino acids.

25. Clusin and yatein and/or dihydrocubebin for use in the treatment or prevention of a tumour disease, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer

26. Clusin, dihydrocubebin and/or hinokinin for use in the treatment or prevention of a tumour disease, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer.

27. Clusin, dihydrocubebin and/or cubebin for use in the treatment or prevention of a tumour disease, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer.

28. Clusin, dihydrocubebin, yatein and/or hinokinin for use in the treatment or prevention of a tumour disease, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-dependent cancer, most preferably for use in the treatment or prevention of prostate cancer.

29. Dihydrocubebin, yatein and/ or hinokinin for use in the treatment or prevention of a tumour disease, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-independent cancer, most preferably for use in the treatment or prevention of prostate cancer.

30. Clusin, dihydrocubebin, yatein and hinokinin for use in the treatment or prevention of a tumour disease, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-independent cancer, most preferably for use in the treatment or prevention of prostate cancer

31 . Clusin, dihydrocubebin, yatein, hinokinin and cubein for use in the treatment or prevention of a tumour disease, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-independent cancer, most preferably for use in the treatment or prevention of prostate cancer.

32. Process for preparing a lignan composition comprising a lignan concentration of at least 60% (w/w), preferably at least 65% (w/w), more preferably at least 70% (w/w) from lignan- containing plant material comprising the steps of:

(a) contacting lignan-containing plant material with an extraction solvent to obtain an extraction solution, wherein the extraction solvent is preferably a supercritical gas or an organic solvent, preferably alcohol, more preferably ethanol,

(b) separating the plant material solids from said extraction solution, preferably comprising filtration or centrifugation,

(c) optionally concentrating said extraction solution, preferably by evaporation or precipitation,

(d) fractionating the extraction solution, preferably by chromatography, more preferably by reversed phase chromatography and/or liquid/liquid fractionation and/or adsorption chromatography,

(e) pooling lignan-containing fractions, and

(f) optionally concentrating the pooled lignan-containing fractions to obtain a lignan composition, wherein the lignan concentration in the lignan composition, preferably the lignan concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin, is at least 60% (w/w), preferably at least 65% (w/w), more preferably at least 70% (w/w).

33. The process according to claim 32, wherein isocratic conditions are used for elution in fractionation step (d) using chromatography, preferably using an alcohol/water solution.

34. The process according to claim 32 or 33, wherein the lignan-containing plant material is derived from plan species selected from the group consisting of Piper cubeba, such as Piper cubeba L., Tarenna madagascariensis, Piper ribesioides, Piper Clusii, Juniperus phoenicea, such as Moroccan Juniperus phoenicea, Juniperus thurifera, such as Juniperus thurifera var. Africana, Chamaecyparis obtuse, and Lychnophora ericoides.

35. The process according to any one of claims 32-34, wherein the plant material derived from Piper cubeba is Piper cubeba L. fruits, preferably unripe Piper cubeba L. fruits as defined in " Deutsches Arzneibuch 6' (DAB6).

36. The process according to any one of claims 32-35, wherein the lignan-containing plant material for preparing the lignan composition are fruits, preferably unripe fruits, from a Piper cubeba L. plant, and wherein the process comprises the steps of:

(a) extracting Piper cubeba powder, preferably C0₂-defatted Piper cubeba powder, with 90%-100% (v/v) ethanol in a drug solvent ratio of 1 :5 (w/w) by agitating the mixture for 1 -4 hours, preferably 2 hours, at between about 15-30°C, preferably at between about 20-25°C,

(c) concentrating the extract by evaporation, preferably on a rotary evaporator, preferably to at least 1/10 to the volume of the extraction solution obtained in step (b),

(d) fractionating the extraction solution comprising the steps:

(d1 ) dissolving the concentrated extract in alcohol, preferably in ethanol, preferably in absolute ethanol,

(d2) subjecting the solution obtained in step (d1 ) to medium pressure liquid chromatography using reverse phase conditions, preferably using polystyrene, silica gels, aluminium oxides, dextrans etc., preferably using RP-C18 silica (e.g. 50 pm), (d3) eluting under isocratic conditions, preferably using a methanol/water solution, preferably at a ratio of methanol : water of 70/30 (v/v),

(d4) collecting fractions of eluted material, preferably at a fraction size of between about 10-100 ml, preferably at about 50 ml,

(e) pooling lignan-containing factions, and

(f) concentrating the pooled lignan-containing fractions, preferably by evaporation, more preferably by freeze drying

to yield the lignan composition.

37. The process according to any one of claims 32-36 further comprising the steps of:

(g) adding one or more pharmaceutically acceptable excipients, diluents and/or carriers to the lignan composition obtained in step (e) or (f),

(h) optionally formulating the lignan composition obtained in step (e), (f) or (g) to obtain a pharmaceutical formulation, preferably a parenteral or non-parenteral formulation.

38. Lignan composition obtainable by the method according to any one of claims 32-37.

39. Pharmaceutical composition comprising the lignan composition according to claim 38.

40. The lignan composition according to claim 38 or the pharmaceutical composition according to claim 39 for use as a medicament.

41 . The lignan composition or the pharmaceutical composition according to any one of claims 38-40 for use in the treatment or prevention of a tumour disease.

42. The lignan composition or the pharmaceutical composition according to claim 41 , wherein the tumour disease is cancer.

43. The lignan composition or the pharmaceutical composition according to claim 41 or 42, wherein the tumour disease or cancer is hormone-dependent, preferably estrogen- or androgen-dependent, such as hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone-dependent colorectal cancer, and hormone-dependent bladder cancer.

44. The lignan composition or the pharmaceutical composition according to any one of claims 38-43 for use in the treatment or prevention of a tumour disease, preferably for use in the treatment or prevention of cancer, more preferably for use in the treatment or prevention of a hormone-independent cancer, most preferably for use in the treatment or prevention of prostate cancer.

45. Composition comprising at least three, preferably at least four, most preferred at least five lignans selected from the group comprising clusin, yatein, hinokinin, dihydrocubebin and cubebin for use in aiding in the prophylaxis or prevention of a tumour disease, preferably for use in aiding in the prophylaxis or prevention of cancer, more preferably for use in aiding in the prophylaxis or prevention of a hormone-independent cancer, most preferably for use in aiding in the prophylaxis or prevention of prostate cancer, wherein

clusin is present in a concentration of at least 10% (w/w), preferably in a concentration of between 10% and 30% (w/w), and/or

dihydrocubebin is present in a concentration of at least 5% (w/w), preferably in a concentration of between 5% and 20% (w/w), and/or

hinokinin is present in a concentration of at least 15% (w/w), preferably in a concentration of between 15% and 50% (w/w), and/or

yatein is present in a concentration of at least 2% (w/w), preferably in a concentration of between 2% and 15% (w/w), and/or

cubebin is present in a concentration of at least 25% (w/w), preferably in a concentration of between 25% and 60% (w/w).

46. Composition according to claim 45, wherein the concentration of lignan compounds together, preferably the concentration of the lignan compounds clusin, dihydrocubebin, hinokinin, yatein and cubebin together is at least 60% (w/w), preferably at least 65% (w/w), more preferably at least 70% (w/w).

47. Composition comprising the lignan composition obtainable according to any one of claims 32 - 37 for use in aiding in the prophylaxis or prevention of a tumour disease, preferably for use in aiding in the prophylaxis or prevention of cancer, more preferably for use in aiding in the prophylaxis or prevention of a hormone-independent cancer, most preferably for use in aiding in the prophylaxis or prevention of prostate cancer.

48. Composition according to any one of claims 45-47, wherein wherein clusin is present in the composition in a concentration of at least 10% (w/w), preferably in a concentration of between 10% and 30% (w/w).

49. Composition according to any one of claims 45-48, wherein yatein is present in a concentration of at least 2%, preferably in a concentration of between 2% (w/w) and 1 5% (w/w).

50. Composition according to any one of claims 45-49, wherein hinokinin is present in a concentration of at least 15% (w/w), preferably in a concentration of between 1 5% (w/w) and 50% (w/w).

51 . Composition according to any of claims 45-50, wherein dihydrocubebin is present in a concentration of at least 5% (w/w), preferably in a concentration of between 5% (w/w) and 20% (w/w).

52. Composition according to any one of claims 45-51 , wherein cubebin is present in a concentration of 25% (w/w) / preferably in a concentration of between 25% (w/w) and 60% (w/w).

53. Composition according to any of claims 45-52, optionally comprising one or more pharmaceutically acceptable excipients, carriers, and/or diluents.

54. Method of treating or preventing a tumour disease comprising administering to a subject in need thereof

(i) a therapeutically effective amount of a pharmaceutical composition according to any one of claims 1 -3, 9-24 and 39,

(ii) a therapeutically effective amount of the lignan composition according to claim 38, or

(iii) a therapeutically effective amount of one or more compounds selected from the group consisting of clusin, dihydrocubebin, hinokinin, yatein and cubebin, preferably selected from the group consisting of clusin, dihydrocubebin and hinokinin, more preferably selected from the group consisting of clusin and dihydrocubebin.

55. The method according to claim 54, wherein the tumour disease is cancer.

56. The method according to claim 54 or 55, wherein the tumour disease or cancer is hormone-dependent, preferably estrogen- or androgen-dependent, such as hormone-dependent breast cancer, hormone-dependent endometrial cancer, hormone-dependent prostate cancer, hormone-dependent testicular cancer, hormone-dependent ovarian cancer, hormone- dependent colorectal cancer, and hormone-dependent bladder cancer.

57. The method according to any one of claims 54-56 for treating or preventing prostate cancer, breast cancer or pancreatic cancer, preferably for treating or preventing prostate cancer, more preferably for treating or preventing hormone-dependent prostate cancer.

58. The pharmaceutical composition according to any one of claims 1 -24 and 39-44, the compound(s) for use according to any one of claims 25-31 the lignan composition according to any one of claims 38 and 40-44, or the method according to any one of claims 54-57, wherein the pharmaceutical composition, the lignan composition or the compound(s) are formulated in a pharmaceutical formulation, preferably in a parenteral or non-parenteral formulation, preferably for oral or intra-tumoural administration.

59. Composition according to any one of claims 38 and 45-53 for use as a health or dietary supplement.

60. Use of a composition according to any of claims 38 and 45-53 for the preparation of a nutritional product.