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					(12)                                       EUROPEAN PATENT SPECIFICATION
(45)       Date of publication and announcement of the              (51)    Int CI.:
                                                                                        (200601)
           advice on the granting of a patent:                              A61K 36/185
           13.06.2007 Patent Bulletin 2007/24
                                                                    (86)    International application number:
(21)       Application number: 02777223.5                                   PCT/EP2002/010813

(22)       Date of application: 26.09.2002                          (87)    International publication number:
                                                                            WO 2003/028746 (10.04.2003 Gazette 2003/15)


(54)         METHOD FOR PRODUCING EXTRACTS OF PELARGONIUM SIDOIDES AND/OR
             PELARGONIUM RENIFORME

             VERFAHREN ZUR HERSTELLUNG VON EXTRAKTEN AUS PELARGONIUM SIDOIDES
             UND/ ODER PELARGONIUM RENIFORME

             PROCEDE D'OBTENTION D'EXTRAITS                                DE      PELARGONIUM            SIDOIDES         ET/OU
             PELARGONIUM RENIFORME


(84)        Designated contracting states:                                        • KOCH, Egon
            AT BE BG CH CY CZ DE DK EE ES Fl FR GB GR                               76229 Karlsruhe (DE)
            IE IT LI LU MC NL PT SE SK TR
                                                                           (74)     Representative: Adam, Holger et al
(30)        Priority: 27.09.2001      DE 10147644                                   Kraus & Weisert,
                                                                                                       attorneys
                                                                                    Patent lawyers and attorneys,
(43)        Date of publication of the application:                                 Thomas-Wimmer-Ring 15
            23.06.2004 Patent Bulletin 2004/26                                      80539 Munich (DE)

(73)        Proprietor of the patent: Dr. Willmar Schwabe                  (56)     Opposition:
            GmbH & Co. KG                                                                                    PELARGONIUM
                                                                                  • KOLODZIEJ H. ET AL: "PELARGONIUM
            76227 Karlsruhe (DE)                                                    SIDOIDES DC." ZEITSCHRIFT FÜR
                                                                                    PHYTOTHERAPIE , Vol. 19, no. 3,1998,
                                                                                                151,
                                                                                    Pages 141-151, XP008011448
(72)        Inventor:                                                               HAIDVOGL M. ET AL: "AKUTE
    •       ERDELMEIER, Clemens                                                     BRONCHITIS IM KINDESALTER."
            76139 Karlsruhe (DE)                                                    ZEITSCHRIFT FÜR PHYTOTHERAPIE,
       •    HAUER, Hermann                                                                                       (1996
                                                                                    Vol. 17, no. 5, October 1996 (1996-10),
            76228 Karlsruhe (DE)                                                                313,
                                                                                    Pages 300-313, XP008011447




             Note: Within nine months from the publication of the mention of the grant of the European patent, any person may
             give notice to the European Patent Office of opposition to the European patent granted. Notice of opposition shall be
             filed in a written reasoned statement. It shall not be deemed to have been filed until the opposition fee has been paid
             (Art. 99(1) European Patent Convention).




                                                 Printed by Jouve, 75001 PARIS (FR)
                                                   EP 1 429 795 B1

     Description

     [0001] This invention concerns a process for the production of Pelargonium sidoides and/or Pelargonium reniforme
     extracts.
5    [0002] Pelargonium sidoides is a plant that has traditionally been used in southern Africa for a long time, as
     medication for gastrointestinal complaints and respiratory diseases, including tuberculosis. The Englishman
     Stevens discovered the medicinal drug in 1897 and brought it to England. The observed treatment successes led to
     the drug being introduced in Switzerland by Sechehaye, for therapeutic purposes. It was used especially in the form
10   of homeopathic preparations for multi-week regimens for tuberculosis prevention in predisposed children and
     adults (Kolodziej, H., Kayser, O., Z. Phytother. 19, 141 - 151, (1998)).
     [0003] Other pharmacological properties of Pelargonium sidoides include general antibacterial effects against
     gram-positive (Staphylococcus aureus, Streptococcus pneumoniae, ß-hemolysing streptococci) and
     gram-negative germs (E. coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeroginosa, Haemophilus
     influenzae), (Kayser, O., Kolodziej, H., Planta Med. 63, 508 - 510 (1997)).
15   [0004] The Pelargonium sidoides species had for a long time been classified as a natural variety of Pelargonium
     reniforme with respect to its systematic botany. A clear separation between Pelargonium sidoides and Pelargonium
     reniforme has for this reason never been made in the older literature (Kolodziej, H., Kayser, O., Z. Phytother. 19,
     141 -151, (1998)). Based on the great botanical similarity between Pelargonium sidoides and the related species
     Pelargonium reniforme, it must be assumed that the roots of Pelargonium sidoides, which generally originate from
20   collections in the wild, contain varying proportions of Pelargonium reniforme.
     [0005] Proanthocyanidins are the main ingredients in Pelargonium sidoides roots, constructed mainly of Catechin
     and Gallocatechin units (Kayser, O., dissertation, Fachb. Pharm. Freie Univ. (FU) Berlin, pp. 23 - 24 and 51 - 55
     (1997)). Simple cumarines are furthermore present, with a generally high degree of oxygenation of the cumarin
     base structure. This appears to be a relatively typical structural characteristic of Pelargonium (Kayser, O.,
25   Kolodziej, H., Phytochemistry 39, 1181 -1185 (1995); Latte, K. P., Kayser, O., Tan, N., Kaloga, M., Kolodziej, H., Z.
     Naturforsch. 55c, 528 - 533 (2000)).
     [0006] Extracts from the Pelargonium sidoides and/or reniforme roots were generally produced, for instance, by
     simple maceration, using acetone / water 4/1 (Kayser, O., Kolodziej, H., Phytochemistry 39, 1181-1185 (1995)) or
     pure methanol using the Soxhlet method (Bladt, S.; Wagner, H. Dtsch. Apoth.-Ztg. 128,292 - 296 (1988).
30   Pure aqueous extracts were also used for therapeutic purposes (Kolodziej, H., Kayser, O., Z. Phytother.
     19,141-151 (1998)). An ethanolic extract of Pelargonium reniforme/sidoides, diluted with glycerine, is available in
     the trade (Red List 2001).
     [0007] The extraction methods used up to now have disadvantages of relatively low yields or serious temperature
     stress (Soxhlet).
35   [0008] It is the purpose of this invention to provide an improved, gentle method for the production of Pelargonium
     sidoides and Pelargonium reniforme extracts, with a higher yield of extracts and with simultaneously also
     enhancing the effect of the extracts.
     [0009] To solve this problem, this invention discloses a process for the production of a Pelargonium sidoides and/or
     Pelargonium reniforme extract, characterised in that the Pelargonium sidoides and/or reniforme roots are subjected
40   either to percolation or two-step maceration.
     The percolation or two-step maceration is preferably performed using aqueous ethanolic solvents. A preferred
     solvent is 10-92 % by weight ethanol and in particular 10-60 % by weight ethanol. Percolation may also be
     achieved by mashing the roots with the solvent, whereby mashing and the subsequent actual percolation may be
     by the same or a different concentration of aqueous ethanol.
45   In the latter case, mashing is preferably with a relatively higher concentration of aqueous ethanol, such as 35 % by
     weight ethanol and the subsequent percolation with a relatively lower concentration of aqueous ethanol, such as
     5 % by weight ethanol, for instance in a ratio of 2:8, to ensure that the average concentration of the ethanol used is
     in the specified ranges (10-92 % by weight, preferably 10-60 % by weight). In the above example, the weighted
     average concentration is 11 % by weight ethanol ([35 x 2 + 5 x 8]/10 % by weight).
50   The extracts obtained in accordance with the invention have a total phenol content of at least 15 %, preferably at
     least 18 %. Such extracts are particularly well suited for the treatment of acute and chronic inflammatory diseases
     and infections.
     [0010] The dry extract yields with percolation in accordance with the invention (examples 1 to 4) and with the
     two-step maceration in accordance with the invention with 11 to 60 % by weight ethanol (examples 5 to 8), are
     about twice as high compared to simple maceration in accordance with the stipulation of the DAB [German
55   Pharmacopoeia] (comparative example 1).
     [0011] In extraction tests with dried Pelargonium sidoides roots and using water-ethanol mixtures, the surprising
     result was that, as the ethanol content in the extraction medium increased, the total phenolic content in the resultant
     extract increased steadily without, as expected, reaching a maximum at medium ethanol concentrations.




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                                                  EP 1 429 795 B1


     The total phenolic content was increased as compared to simple maceration (comparative example 1), both with
     the percolation (examples 1 to 4) and with the two-step maceration (examples 5 to 9) by a factor of up to 3,
     depending on the extraction solvent medium.
     [0012] The anti-oxidative potential of the extracts increases in parallel with the total phenolic content.
5    In this respect, the extracts in accordance with the invention are superior to the comparative extracts in accordance
     with the comparative examples 1 and 2. Since the total cumarin content in the extracts remains virtually constant
     irrespective of the process and the solvent, it must be assumed that the contribution the cumarins are making to the
     anti-oxidative effect is insignificant.
     [0013] When comparing examples 1 (mashing with 35 % by weight ethanol and percolating with 5 % by weight
     ethanol), 2 (mashing with 11 % by weight ethanol and percolating with 11 % by weight ethanol) and 4 (percolating
10   with 11 % by weight ethanol without prior mashing), we find the highest yield for the extract in accordance with
     example 1 and the highest total phenolic content for the extract in accordance with example 4. For the extracts in
     accordance with examples 1 and 4, the anti-oxidative potential is somewhat higher than that of the extracts in
     accordance with example 2. The IC50 values for elastase inhibition are also virtually the same for the extracts in
     accordance with examples 1 and 4, but the IC50 value for the extract in accordance with example 2 is comparatively
15   high at 8.4 µg/ml. The processes in accordance with examples 1 and 4 are therefore preferable to the process in
     accordance with example 2. Percolation without mashing and percolation with mashing using higher
     concentrations of ethanol and percolation with less concentrated ethanol are therefore preferable to a process in
     which mashing and percolation are done with the same concentrations of ethanol.
     [0014] The commercially available preparation of Pelargonium reniforme/sidoides is administered especially for the
     treatment of acute and chronic infections of especially the respiratory tracts and the ear, nose and throat regions.
20   Anti-microbial action and/or the stimulation of unspecific immune responses are considered the main
     pharmacological effects of the product. Neutrophile granulocytes are first in the line of defence against pathogens
     and especially bacteria. The stimulation of these leucocytes, e.g. in the course of phagocytosis of microorganisms,
     is accompanied by increased oxygen consumption and the formation of highly reactive oxides (e.g. superoxide,
25   hydrogen peroxide) which assist these cells in killing infectious agents. If the defensive reaction overshoots and in
     the course of chronic infections, the reactive oxygen species set free by the phagocytosing cells can, however, also
     cause tissue damage. Substances with anti-oxidative properties can prevent these undesirable side effects,
     thereby improving and accelerating the healing process (Cuzzocrea et al., Pharmacol. Rev. 53, 135 (2001)). Since
     the anti-oxidative properties are dependent on the total phenolic content, the extracts in accordance with the
30   invention are therefore clearly superior to comparative extracts.
     [0015] Apart from reactive oxygen species, the phagocytosing cells also make use of proteolytic enzymes to kill
     bacteria. Elastase is one of the most important and potent proteases formed and released by the leucocytes. Due
     to its destructive effect on the host tissue, this enzyme has pathophysiological significance and elastase inhibitors
     therefore have broad therapeutic applications in cases of acute and chronic inflammatory diseases such as
35   bronchitis and other respiratory tract diseases, lung emphysema, cystic fibrosis, rheumatoid arthritis, myocarditis,
     etc. (Fujie et al., Eur. J. Pharmacol., 374, 117 (1999)).
     [0016] In a surprising development, it has now been observed that extracts of Pelargonium sidoides and/or
     reniforme possess potent inhibitory qualities with respect to human leucocyte elastase (HLE) and that this effect is
     also correlated with the quality of the total phenolic extracts.
40   The dual effective mechanism of anti-oxidative properties and inhibition of the HLE is considered particularly
     advantageous since it actively affects two of the most important pathogenetic factors contributing to body tissue
     damage as a result of chronic inflammatory processes (Portevin et al., J. Med. Chem. 40, 1906 (1997)).
     [0017] Together with the conventional pharmaceutical excipients, the obtained extracts can be processed into
     pharmaceutical solid forms such as capsules, tablets, coated tablets or chewing tablets.
45   The pharmaceutical excipients used are conventional fillers, binders, disintegrants and lubricants as well as
     scenting and flavouring agents and coating substances for coated tablets. Conventional oils and fats are used as
     fillers for soft gelatine capsules as pharmaceutical excipients in the manufacture of soft gelatine capsules. The
     extracts obtained can be processed into pharmaceutical liquid preparations such as solutions, sprays and
50   suspensions together with conventional pharmaceutical excipients. The pharmaceutical excipients are
     conventional solvents, solubilisers, stabilisers and scenting and flavouring agents.
     [0018] The invention is explained with reference to the following non exclusive examples. To obtain comparable
     analytical and pharmacological data, all the examples were dried to obtain the dry extract. The dry extracts do not,
     however, represent the underlying extracted solutions, which are also a subject matter of the invention.

55




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                                                  EP 1 429 795 B1


     Examples 1 to 4 – Percolation with different extraction solvents

     Example 1

5    [0019] 1 kg dried Pelargonium sidoides roots were soaked with 2 kg 35 % by weight EtOH for 24 hours to swell and
     then transferred to a glass column. Percolation was then performed for 10 h with 8 kg 5 % by weight EtOH and the
     drug residue thereafter slightly squeezed, the raw extract filtered through Seitz Supra 1500 and concentrated to
     dryness.

     Examples 2 and 3
10
     [0020] 40 g dried Pelargonium sidoides roots were soaked with 80 g extraction solvent (11 % and 35 % by weight
     ethanol resp.) for 22 hours to swell and then transferred to a glass column. Percolation was then performed for 3 to
     4 h with 320 g of the above extraction solvent and thereafter the drug residue was slightly squeezed and the clear
     raw extract concentrated to dryness.
15
     Example 4

     [0021] 40 g dried Pelargonium sidoides roots were transferred into a glass column and percolated for 8 h using
     400 g 11 % by weight ethanol. The drug residue was thereafter slightly squeezed, the raw extract filtered through
20   Seitz Supra 1500 and concentrated to dryness.

       Example no.       Extraction  Yield         Total phenolic    Total cumarins Antioxidative      Elastase
                         solvent     (%)           content           (%)            potential µg       inhibition
25                       % by weight               (%)                              Trolox / mg        IC50 (µg/ml)
                         EtOH
                         11 (35%/
       1                               18.4             17.47               3.18      279                    5.7
                         5%)
       2                 11            15.3             17.09               3.14      237                    8.4
30     3                 35               17.2          24.16               2.84      311                    6.2
       4                 11               17.8          21.79               2.69      273                    5.9

     Comparative example 1 – Maceration in acc. with DAB [German Pharmacopoeia]
35
     [0022] 20 g dried Pelargonium sidoides roots were extracted using 140 ml 11 % by weight ethanol at room
     temperature for 5 days with occasional shaking and subsequent storage for 5 days at a cool temperature, in
     accordance with the maceration instructions for extracts in DAB 10. Thereafter, filtration was carried out and the
     extract solution was dried.
40
     Examples 5 to 9 and comparative example 2 – Maceration with different extraction solvents

     [0023] 20 g dried Pelargonium sidoides roots were macerated using 140 g extraction solvent (ethanol / water in the
     ratio shown below or in water (comparative example 2)) at 50 °C for 30 minutes. The extract solution is thereafter
45   filtrated off and the drug residue macerated a second time in the same way. The extract solutions were combined
     and dried after the solid / liquid separation (filtration).

        Example no.      Extraction  Yield         Total phenolic       Total cumarins Antioxidative   Elastase
50                       solvent     (%)           content              (%)            potential µg    inhibition
                         % by weight               (%)                                 Trolox / mg     IC50 (µg/ml)
                         EtOH
        Comparison 1     11          8.0                 12.37               3.14      211             >10
55      Comparison 2     0 (water)      13.8             14.07               2.69      234             9.0
        5                11             16.2             18.72               2.65      271             6.0




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                                                   EP 1 429 795 B1


                                                       (continued)

         Example no.      Extraction  Yield         Total phenolic    Total cumarins Antioxidative      Elastase
                          solvent     (%)           content           (%)            potential µg       inhibition
5                         % by weight               (%)                              Trolox / mg        IC50 (µg/ml)
                          EtOH
         6                20             16.5             20.10             2.42       304              5.0
         7                35             17.6             24.13             2.43       353              4.2
10       8                60             15.7             27.53             2.63       366              5.9
         9                90             7.6              36.18             2.93       508              4.5

     Analytical and pharmacological methods
15

     Determining the total phenolic content

     [0024] The total phenolic content is determined photometrically after conversion using molybdate tungstate
     reagent analogous to the Pharmacopoeia method for tanning agents (DAB 2000). The extract is, for this
20   purpose, dissolved in aqueous ethanol, alkalised with sodium carbonate solution and added with molybdate
     tungstate reagent. After centrifuging, the extinction of the supernatant solution is measured at 720 nm against
     water. The calculation is on epicatechin.

     Determining the total cumarin content

25   [0025] The total cumarin content is determined with HPLC via a RP-18 column. An acetonitrile / water / phosphoric
     acid gradient (10:990:4 → 205:795:4) is used as the mobile phase. Detection is at 330 nm UV. The individual
     cumarin peaks are calculated as scopoletin.

     Determining the anti-oxidative potential
30
     [0026] The Shi, H. and Niki, E. (Lipids 33, 365 (1998)) method used is based on the photometric measurement of
     the extinction change associated with the reduction of a stable radical (galvinoxyl). For this purpose, 2.69 mg
     galvinoxyl are dissolved in 500 ml ethanol (12.75 µmol/l). 8 ml of this are added with 200 µl sample or calibration
     solution (see below) and incubated in the dark for 20 min. at room temperature. Thereafter, measure at 428 nm
35   with the photometer.
     Calibration solution: 0.954 mg Trolox (6-Hydroxy-2,5,7,8 Tetramethylchroman-2-Carboxylic Acid) are dissolved in
     25 ml ethanol / water 1/1. This master solution is used to make 1/25,1/20,1/10,1/5 and 1/2 dilutions in
     ethanol / water 1/1 and the absorption measured at 428 nm.

40   Determining the elastase inhibition

     [0027] The effect of extracts on the peptidolytic activity of HLE (E.C. 3.4.21.37; Sigma) was determined at room
     temperature in 50 mM Tris-HCI (pH 8) with 50 mM NaCI and 0.01 % Brij 35. HLE (50 µl, 200 mU/ml) was incubated
     for 15 minutes with 50 µl of the extracts in a microtitre plate. The enzyme reaction was thereafter started by addition
45   of 50 µl substrate (Methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide; 1 mM; Sigma) and measured in a microplate
     photometer (Thermomax, Molecular Devices) at 405 nm over a 10 min. period. The inhibition of the enzyme activity
     was calculated by comparison with an identical solvent control (DMSO).

50   Claims

     1   Method for producing an extract from Pelargonium sidoides and/or Pelargonium reniforme, characterised in that
         the roots of Pelargonium sidoides and/or reniforme are either

             a)   subjected to percolation using an aqueous ethanolic solvent, the drug residue is optionally slightly
                  squeezed and the raw extract is optionally filtered, or
             b)   subjected to a two-step maceration using an aqueous ethanolic solvent, wherein the extract solution is
                  filtered after the first maceration and the drug residue is macerated a second time and the extract
                  solutions are combined after a solid/liquid separation.




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                                              EP 1 429 795 B1



2   Method according to claim 1, wherein the liquid extract obtained in step a) or b) is dried to obtain a dry extract.

3   Method according to claim 1 or 2, wherein an initial mashing using an aqueous ethanolic solvent is carried out
    before the actual percolation step in the case of the percolation.

4   Method according to claim 3, wherein the initial mashing and the percolation are carried out using aqueous
    ethanol having different concentrations.

5   Method according to any one of Claims 1 to 3, wherein the aqueous ethanolic solvent is 10-92% by weight
    ethanol.

6   Method according to any one of Claims 1 to 3, wherein the aqueous ethanolic solvent is 10-60% by weight
    ethanol.

7   Method according to claim 4, wherein the weighted average concentration of the aqueous ethanol used for
    initially mashing and percolating is within a range of 10-92% by weight, preferably 10-60% by weight.

8   Method according to claim 4, wherein 35% by weight ethanol is used for initially mashing and 5% by weight
    ethanol is used for percolating in a ratio of 2:8.

Patentansprüche

1   Verfahren zur Herstellung eines Extraktes aus Pelargonium sidoides und/oder Pelargonium reniforme, dadurch
    gekennzeichnet, dass die Wurzeln aus Pelargonium sidoides und/oder reniforme entweder

       a)   einer Perkolation mit einem wässrig-ethanolischen Lösungsmittel unterworfen werden, der
            Drogenrückstand gegebenenfalls leicht ausgepresst und der rohe Extrakt gegebenenfalls filtriert wird,
            oder
       b)   einer zweistufigen Mazeration mit einem wässrig-ethanolischen Lösungsmittel unterworfen werden,
            wobei nach der ersten Mazeration die Extraktlösung abfiltriert und der Drogenrückstand ein zweites
            Mal mazeriert wird und die Extraktlösungen nach der Fest/Flüssigtrennung vereinigt werden.

2   Verfahren nach Anspruch 1, wobei der in Schritt a) oder b) erhaltene flüssige Extrakt getrocknet wird, um
    einenTrockenextrakt zu erhalten.

3   Verfahren nach Anspruch 1 oder 2, wobei bei der Perkolation vor dem eigentlichen Perkolieren ein Anmaischen
    mit einem wässrig-ethanolischen Lösungsmittel erfolgt.

4   Verfahren nach Anspruch 3, wobei das Anmaischen und das Perkolieren mit unterschiedlich konzentriertem
    wässrigen Ethanol erfolgt.

5   Verfahren nach einem der Ansprüche 1 bis 3, wobei das wässrig-ethanolische Lösungsmittel 10-92 Gew.-%
    Ethanol ist.

6   Verfahren nach einem der Ansprüche 1 bis 3, wobei das wässrig-ethanolische Lösungsmittel 10-60 Gew.-%
    Ethanol ist.

7   Verfahren nach Anspruch 4, wobei die gewichtete durchschnittliche Konzentration des beim Anmaischen und
    beim Perkolieren verwendeten wässrigen Ethanols im Bereich von 10-92 Gew.-%, vorzugsweise 10-60
    Gew.-% liegt.

8   Verfahren nach Anspruch 4, wobei für das Anmaischen 35 Gew.-% Ethanol und das Perkolieren 5 Gew.-%
    Ethanol im Verhältnis 2:8 verwendet wird.


Revendications

1   Procede de production d'un extrait de Pelargonium sidoides et/ou de Pelargonium reniforme, caracterise en ce
    que les racines de Pelargonium sidoides et ou reniforme




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                                              EP 1 429 795 B1


       a)   asont soumises ä une percolation avec un solvant aqueux ethanolique, le residu medicamenteux est
            even-tuellement legerement presse et l'extrait brut est eventuellement filtre, ou
       b)   asont soumises ä une maceration en deux etapes avec un solvant aqueux etanolique, la Solution
            d'extraction etant filtree apres la premiere maceration et on fait macerer une deuxieme fois le residu
            medicamenteux et les Solutions d'extraction sont reunies apres la Separation en phases solide/liquide.

2   Procede selon la revendication 1, l'extrait fluide obtenu ä l'etape a) ou ä l'etape b) etant sechepourobtenirun
    extrait sec.

3   Procede selon la revendication 1 ou 2, un foulage avec un solvant aqueux ethanolique avant la percolation
    effective etant effectue lors de la percolation.

4   Procede selon la revendication 3, le foulage et la percolation s'effectuant avec de l'ethanol aqueux ä differentes
    concentrations.

5   Procede selon l'une quelconque des revendications 1 ä 3, le solvant aqueux ethanolique etant de l'ethanol de ä
    92 % en poids.

6   Procede selon l'une quelconque des revendications 1 ä 3, le solvant aqueux ethanolique etant de l'ethanol de ä
    60 % en poids.

7   Procede selon la revendication 4, la concentration moyenne ponderee de l'ethanol aqueux utilise lors du
    foulage et de la percolation se situant dans une plage allant de 10 ä 92 % en poids, de preference, de 10 ä 60 %
    en poids.

8   Procede selon la revendication 4, sachant que l'on utilise pour le foulage, de l'ethanol ä 35 % en poids et pour la
    percolation, de l'ethanol ä 5 % en poids en un rapport de 2 : 8.




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                                            EP 1 429 795 B1


DOCUMENTS REFERENCED IN THE DESCRIPTION

This list of documents referenced by the applicant was compiled exclusively to serve as information for the reader
and is not a component of the European patent document. The list was compiled with great care; however, the EPO
accepts no liability for any errors or omissions.

Non-patent literature referenced in the description

•    KOLODZIEJ, H ; KAYSER, 0., Z. Phytother, 1998, •           KAYSER, 0; KOLODZIEJ, H. Phytochemistry,
     vol. 19, 141-151 [0002] [0004] [0006]                      1995, vol. 39, 1181-1185 [0006]
•    KAYSER, 0 ; KOLODZIEJ, H. Planta Med., 1997, vol. •        BLADT, S ; WAGNER, H. Dtsch. Apoth.-Ztg,
     63, 508-510 [0003]                                         1988, vol. 128, 292-296 [0006]
•    KAYSER, O. Dissertation, Fachb. Pharm. Freie Univ, •       CUZZOCREA et al. Pharmacol. Rev, 2001, vol.
     1997, 23-2451-55 [0005]                                    53, 135 [0014]
•    KAYSER, O; KOLODZIEJ, H. Phytochemistry, 1995, •           FUJIE et al. Eur. J. Pharmacol., 1999, vol.
     vol. 39, 1181-1185 [0005]                                  374,117 [0015]
•    LATTE, K. P ; KAYSER, O ; TAN, N ; KALOGA, M ; •           PORTEVIN et al. J. Med. Chem., 1997, vol.
     KOLODZIEJ, H., Z. Naturforsch., 2000, vol. 55c,            40,1906 [0016]
     528-533 [0005]                                         •   SHI, H ; NIKI, E. Lipids, 1998, vol. 33, 365 [0026]




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