CA1050427A - Pharmaceutical preparation containing the principal helleborus sapogenin - Google Patents
Pharmaceutical preparation containing the principal helleborus sapogeninInfo
- Publication number
- CA1050427A CA1050427A CA224,029A CA224029A CA1050427A CA 1050427 A CA1050427 A CA 1050427A CA 224029 A CA224029 A CA 224029A CA 1050427 A CA1050427 A CA 1050427A
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- roots
- rhizomes
- principal
- helleborus
- sapogenin
- Prior art date
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Abstract
ABSTRACT OF THE DISCLOSURE
A pharmaceutical preparation for the treatment of ulcers includes the principal sapogenin of the roots and rhizomes of the Helleborus species and a pharmaceutically acceptable diluent or carrier. Such principal sapogenin is extracted from the roots and rhizomes by treating the roots and rhizomes with an enzyme selected from the group consisting of cellulase, hemicellulase and .beta.-glucosidase.
A pharmaceutical preparation for the treatment of ulcers includes the principal sapogenin of the roots and rhizomes of the Helleborus species and a pharmaceutically acceptable diluent or carrier. Such principal sapogenin is extracted from the roots and rhizomes by treating the roots and rhizomes with an enzyme selected from the group consisting of cellulase, hemicellulase and .beta.-glucosidase.
Description
lOS042~
This invention relates to a pharmaceutical preparation including sapogenin from roots of the l~elleborus species and a pharmaceutically acceptable carrier.
The roots and rootstocks of Helleborus species contain a mixture of saponins the principal sapogenin of which has the structure of a spirost-5,25~27)-diene-1~, 3~ -triol ~formula I) , 3 CH~ ; ~` ~ CH ( I ) El ", .
According to Helv. Chim. Acta 54, 1707 (1971), pulverized rhizomes (rhizomes are underground, more or less thickened root-ing stems, which can be clearly distinguished from roots by the presence of scale-like lower leaves and by their structure) and ¦ -roots of Helleborus species are coarsely pulverized, taken up in water and left to autoferment for 22 days in order to obtain the above sapogenin. The residue is subsequently filtered and extracted with aqueous alcohol. The residue of the alcoholic extract is chromatographed on silica gel. The sapogenin frac-tions are recrystallized from methanol-acetone. The pure sapo-i , ,.
;~ yenin (m.p. 236-240C) is finally obtained by preparatory thin-layer chromatography in the system diisopropyl ether ethanol ; (92:8).
It has been found that this principal genin of the sapo- F
~;~ genin mixture - which is contained in the roots and rhizomes of Helleborus species - can be obtained in a simpler and faster way : if the roots and rhizomes of Helleborus species or drugs or ~-extracts obtained therefrom are treated with an enzyme or enzyme .
mixture, the important active components of which are cellulase (3.2.1.4), hemicellulase or ~-glucosidase (3.2.1.21). Accordiny - A~
~o J.-E. Courtois alld Mme. Bui Khac Diep ~Annales pharmaceuti~ues francaise 23, 1965, no. 9-10, pp.533-459) hemmicellulase is a mixture of ~-galactosidase (3.2.1.22) g-mannanase (3.2.1.25) and ~-mannosidase (3.2.1.25) , ' ~
`: ;
" `, .
, 20 '.
.. _ ', ,, ~.
. ' , . ~ .
~ 30 :
r . , ~.
~ - la - .
':' , 10504~
The treatment with an enzyme is carried out in the usual manner. The temperature is suitablY kept between 20 and 50~C.
A temperature between 30 and ~QC i9 particular]y favourable.
In the above process, the comminuted roots and rhizomes of ~elleborus species can be used directl~ or an extract obtained in the usual manner or a drug obtained from Helleborus species, treated and pulverized in the usual manner, can be used. The treatment can be carried out in solution, in suspension or in the form of a mash. Thus, for example, the enzyme can be added to the autolyzate of roots and rhizomes of Helleborus species.
The autolysis or autofermentation can be effected by comminuting parts of fresh plants and, if necessary, after adding water allow-ing the material thus obtained to stand for a lengthy period (e.g., 2 to 25 days) at a temperature of 18 to 60C, while stirr-ing occasionally. Parts of dried plants can also be mixed with water and subjected to autofermentation as long as their ferment-ative activity is maintained. The enzyme can be added at the start of the autofermentation process or even during said process.
However, it is also possible first to produce an extract in a known manner from the roots and rhizomes or from a drug produced therefrom and treat it with the enzyme, if required, after the usual preliminary purification. For this purpose, the roots and rhyzomes or the drugs are-extracted with alcohol or ., .". ".
~ alcohol-water mixtures containing a maximum of 50~ water. The - extract thus obtained is subse~uently shaken with an organic solvent such as an aromatic hydrocarbon, halohydrocarbon or a mixture of a halohydrocarbon and alcohol such as for example, chloroform-ethanol. If mixtures of a halohydrocarbon and alcohol are used, then the ratio of the mixtures is preferably 2:1. It is favourable to pre~purify the organic extract thus obtained.
This type of preliminary purification is carried out in
This invention relates to a pharmaceutical preparation including sapogenin from roots of the l~elleborus species and a pharmaceutically acceptable carrier.
The roots and rootstocks of Helleborus species contain a mixture of saponins the principal sapogenin of which has the structure of a spirost-5,25~27)-diene-1~, 3~ -triol ~formula I) , 3 CH~ ; ~` ~ CH ( I ) El ", .
According to Helv. Chim. Acta 54, 1707 (1971), pulverized rhizomes (rhizomes are underground, more or less thickened root-ing stems, which can be clearly distinguished from roots by the presence of scale-like lower leaves and by their structure) and ¦ -roots of Helleborus species are coarsely pulverized, taken up in water and left to autoferment for 22 days in order to obtain the above sapogenin. The residue is subsequently filtered and extracted with aqueous alcohol. The residue of the alcoholic extract is chromatographed on silica gel. The sapogenin frac-tions are recrystallized from methanol-acetone. The pure sapo-i , ,.
;~ yenin (m.p. 236-240C) is finally obtained by preparatory thin-layer chromatography in the system diisopropyl ether ethanol ; (92:8).
It has been found that this principal genin of the sapo- F
~;~ genin mixture - which is contained in the roots and rhizomes of Helleborus species - can be obtained in a simpler and faster way : if the roots and rhizomes of Helleborus species or drugs or ~-extracts obtained therefrom are treated with an enzyme or enzyme .
mixture, the important active components of which are cellulase (3.2.1.4), hemicellulase or ~-glucosidase (3.2.1.21). Accordiny - A~
~o J.-E. Courtois alld Mme. Bui Khac Diep ~Annales pharmaceuti~ues francaise 23, 1965, no. 9-10, pp.533-459) hemmicellulase is a mixture of ~-galactosidase (3.2.1.22) g-mannanase (3.2.1.25) and ~-mannosidase (3.2.1.25) , ' ~
`: ;
" `, .
, 20 '.
.. _ ', ,, ~.
. ' , . ~ .
~ 30 :
r . , ~.
~ - la - .
':' , 10504~
The treatment with an enzyme is carried out in the usual manner. The temperature is suitablY kept between 20 and 50~C.
A temperature between 30 and ~QC i9 particular]y favourable.
In the above process, the comminuted roots and rhizomes of ~elleborus species can be used directl~ or an extract obtained in the usual manner or a drug obtained from Helleborus species, treated and pulverized in the usual manner, can be used. The treatment can be carried out in solution, in suspension or in the form of a mash. Thus, for example, the enzyme can be added to the autolyzate of roots and rhizomes of Helleborus species.
The autolysis or autofermentation can be effected by comminuting parts of fresh plants and, if necessary, after adding water allow-ing the material thus obtained to stand for a lengthy period (e.g., 2 to 25 days) at a temperature of 18 to 60C, while stirr-ing occasionally. Parts of dried plants can also be mixed with water and subjected to autofermentation as long as their ferment-ative activity is maintained. The enzyme can be added at the start of the autofermentation process or even during said process.
However, it is also possible first to produce an extract in a known manner from the roots and rhizomes or from a drug produced therefrom and treat it with the enzyme, if required, after the usual preliminary purification. For this purpose, the roots and rhyzomes or the drugs are-extracted with alcohol or ., .". ".
~ alcohol-water mixtures containing a maximum of 50~ water. The - extract thus obtained is subse~uently shaken with an organic solvent such as an aromatic hydrocarbon, halohydrocarbon or a mixture of a halohydrocarbon and alcohol such as for example, chloroform-ethanol. If mixtures of a halohydrocarbon and alcohol are used, then the ratio of the mixtures is preferably 2:1. It is favourable to pre~purify the organic extract thus obtained.
This type of preliminary purification is carried out in
- 2 :
~OS042'7 the usual manner. However, it may~also be carried out by chroma-tography on a silica geI, the silica gel being a synthetically produced, highIy porous a~orphous silica, in the form of hard particles having a granular size of 0.15 to 10 mm. A granular size of 0.15 to 0.30 mm is particularl~ favourable. The water content of this silica can be up to 10~. The specific surface area can he up to 650 sq m per gram. It usually is approximately 4000 sq m per gram. The powder density can be up to 650 g per litre. A powder density of 450 to 500 g per litre is favourable.
Suitable eluants for the silica-gel chromatography of the i organic extract of the roots and rhizomes of ~elleborus species include aliphatic halohydrocarbons, mixtures of aliphatic halo-hydrocarbons with aliphatic alcohols, esters o~ aliphatic acids with aliphatic alcohols, ester-alcohol mixtures, ester-alcohol-water mixtures, benzene, halogen benzenes, alkyl benzenes, alkyl benzene-alcohol mixtures, ester-pyridine mixtures, halohydrocar-bon-pyridine mixtures, halohydrocarbon-alcohol-pyr dine mixtures, ester-pyridine-water mixtures, mixtures of benzene, halogen benzenes and alkyl benzenes with pyridine, aliphatic ketones, ketone-water mixtures, ketone-benzene mixtures, ketone-benzene-acetic acid mixtures, etc. Of course, other mixtures with the ` above components are also possible. The optimum mixing propor-tion of the components must always be determined in a separate preliminary test.
The aliphatic li~uids mentioned above are the usual lower liquid agents used as solvents. In this connection, halogen means fluorine, chlorine or bromine, preferably fluorine or chlorine.
By alkyl benzenes are meant the liquid agents with lower alkyl radicals. Examples are toluene, ethyl henzene, xylenes.
The pre-purified saponin fraction, which is completely free from solvents, is then incubated in water with the enzyme.
.
~ .
~05(~4'~
Toluene can be a~ed (usually in small amounts) to the incubate as a protection against bacterial decomposition.
For the process, commercial cellulase, hemicellulase and ~-glucosidase preparations can be used. Likewise, it is also possible to use preparations of the enzymes produced from fungi, microorganisms (Trichoderma viride), protozoa, bacterial insects, plants, intervertebrates such as snails (edible snails~ and worms by means of the processes known for this purpose [see The Enzymes 1, part 2 (1951) page 729 ff; Ullmann's Enzyklopadie der technis-chen Chemie, 3rd edition (1956) page 391 ff].
The enzymes such as, for example, the cellulase, are obtained by precipitating the culture medium or the aqueous Mycelium extract of the fungi with alcohol, acetone or salt. The crude enzyme can then be further purified, for example, on alu-minium oxide. L
The preparations to be used must be as fresh as possible t without being stored for a lengthy period. Stored preparations must be kept cool and dry. Enzyme preparations or enzyme con-centrates produced from fungi, for example, Aspergillus species such as Aspergillus niger, ~spergillus flavus, Aspergillus oryzae, Aspergillus fumigatus, Aspergillus nidulans are parti-- culary suitable. Preparations or enzyme concentrates produced from Aspergillus niger are particularly favourable.
-; The activity of the enzymes must be determined by pre-liminary tests and the optimum conditions for the reaction and k quantitative relations must also be determined. Enzyme concen-trates which, for example, consist primarily of cellulase, hemi-cellulase or e-glucosidase or of mixtures of such enzymes are particularly favourable. Frequently, such preparations also con- r 30 tain related enzymes such as pectinase (3.2.1.15), amylase (3.2.1.1), acid protoase (3.4.23.6), xylanase (3.2.1.32), cello-biase (3.2.1.21), glucoamylase (3.2.1.3), endopeptidase (3.4.21.16), 5., ' lipase (3.1.1.3), and pectin ~ . . . . . . .
exopolygalacturonase. It is also favourable if the enzvme pre-parations according to the invention also contain enæymes having a macerating effect (for example, ~ectin glvcosidase~. Examples are highIy purified products obtained from Aspergillus species and containing primarily cellulase as hemicellulase as well as pectinase, amylase and acid protease (cellulosin preparations).
The enzymatic reaction isusuallycompleted after two days. If the activity of the enzymes is high, then the reaction is comple-ted earlier, but, if the activity of the enzymes is low, then the treatment is likely to require a much longer period.
The enzyme can be mixed with the substrate either as such or in aqueous solution.
The optimum pH value for the treatment with the enzymes is from approximately 4.5 to 4.7. It is favourable to stir the incubate or to ~eep it in motion in some other manner.
Acid proteases contain cellulosin preparations.
The amount of the enzyme added depends on the activity of the enzyme and on the substrate used. The enzyme can be added, for example, in substantial excess. For example, if a crude saponin fraction which is pre-puri~ied as described hereinbefore is used, then an addition of 5 to 100~ of the enzyme preparation concerned is required depending on the degree of purity, which is determined by thin-layer chromato~raphy. For example, if the drug or the roots and rhizomes of Helleborus species are reacted directly, then an amount from 1 to 10~, relative to the drug or to the roots and rhizomes should be sufficient.
The crude sapogenin obtained after the enzyme treatment can be further processed by means of the usual process describedin -~ ~heliterature~ For example, the process described in Helv. Chim.
~,~
Acta 54, page 1707 (chromatography on SiO2 with a granular size of 0.05 to 0.2 mm) is suitable for this purpose.
_ 5 ~, .
1~)504Z~
Howe~er! a purification (~or example, by chromatography) of the crude sapoqenin with the aid of a silica gel having a grain size of 0~05 to 0,2 mm is particularly favourable. Lower halogenated hydrocarbons such as dichloromethane and chloroform, to which a lower aliphatic alcohol such as methanol or ethanol can be added, are suitable as eluants, the amount of alcohol increasing from 1%.
The sapogenin thus obtained can be recrystallized once more from propanol/water.
10By means of the above process it is possible to isolate the Helleborus sapogenin in substantially larger amounts than by way of the preparatory thin-layer chromatography.
The known Helleborus species, for example, Hellevorus foetidus L., Helleborus multifidus Vis., Helleborus niger L., Helleborus adorus Waldst. et Kit., Helleborus orientalis Lam., Helleboris purpurascens Waldst. et Kit., Helleborus viridis, are suitable starting materials.
Nothir~was previously known concerning the physiological effect of the principal sapogenin from the roots and rhizomes of Helleborus species. It has now been discovered that the princi-pal genin produces from Helleborus saponin has a curative effect on ulcers. Moreover, a muscle-relaxing effect and an effect influencing the central nervous system have also been detected.
The ulcer-protective effect is evident, for example, from the table hereafter:
Daily Dose Ulcer-Protective Effect :
50 mg per kg 58 100 mg per kg 78 200 mg per kg 80 30These tests were carried out on the indomethacin-induced - ulcer of rats using the method of WilheImi which was modified by .. . .. . . : . . :
~504'~7 Jahn and Adrian [see Arzneimittelfor~chung 19 (1969) page 45 ff].
The determination was carried out in the ~ollowing manner:
Albino rats of the strain SIV 50 (S. Ivanovas, Kissle~g/
Allgau) which had an initial weight of 250 to 300 g were kept in wire cages (Ebeco, type 3, double width) in rooms, the temperature of which was kept between 20 and 22C, on a standard diet (Altromin ~). After withholding food for 48 hours, the animals are given 20 mg of indometh~cin in 1.5% tragacanth (l ml of tragacanth solution per 100 g of rat) per kg of rat, applied intragastrally. One hour thereafter the animals are given the test substance orally. The rats continue to stay sober (water ad libitum).
; 24 hours after administration if indomet~a-cin, the animals are killed by means of CO2. The stomachs are resected, opened along the large curvature and rinsed under running water.
The ulcerative changes appear as dark punctiform or ~-striated spots on the mucosa. The evaluation is carried Ollt macroscopically, using the method of MUNCHOW [Arzneimittelfor-schung 4 (1954) page 341].
Therefore, the present invention provides a pharmaceu-tical preparation containing compound I as the active ingredient, if required, mixed with other pharmacologically or pharmaceuti-cally active substances. The medicines can be produce using `; conventional pharmaceutical adjuvant substances.
~ For example, substances which are recommended and/or ,:
listed in the following references from the literature as adju-vant substances for pharmacy, cosmetics and related fields are .; .
;~ suitable as this kind of fillers and adjuvent substances:
Ullmann's Encyklop~die der technischen Chemie, Vol. 4 (1953), - 30 page 1 to 39; Journal of Pharmaceutical Sciences, Vol. 52 ~1963), - page 918 ff, H`.v. Czetsch-Lindenwald, Hilfsstoffe f~r Pharmazie ,~
; - 7 ,~ .
.
... ~ . , 105042~
und angrenzende Gebiete; Pharm. Ind ~ No. 2, 1961, page 72 ff;
Dr. H.P. Fiedler, Lexikon der Hilfsstoffe f~r Pharmazie, Kosmetik and angrenzende Gebiete~ Cantor RG., ~ulendorf i. W~rtt.
1971.
Examples are gelatin, natural sugar such as sucrose or lactose, lecithin, pectin, starch (for example, cornstarch), tylose, talc lycopodium, silica ~for example, colloidal silica), cellulose, cellulose derivatives (for example, cellulose ethers in which the cellulose-hydroxy groups are partially etherified with lower saturated aliphatic alcohols and/or lower saturated aliphatic oxy-alcohols, for example, methyl-oxy-propyl cellulose), stearates, magnesium and calcium salts of fatty acids, containing 12 to 22 carbon atoms, particularly of the saturated ones (for example, stearates), emulsifiers, oils and fats, particularly vegetable oils (for example, peanut oil, caster oil, olive oil, sesame oil, cottonseed oil, corn oll, mono-, di- and trigl~cer-ides of saturated fatty acids C12H24O2 to Cl8H36o2 mixtures), pharmaceutically compatible monohydric or polyhydric alcohols and polyglycols such as polyethylene glycols as well as derivatives thereof, esters o~ aliphatic saturated or unsaturated fatty acids (2 to 22 carbon atoms, particularly 10 to 18 carbon atoms) with monohydric aliphatic alcohols (1 to 20 carbon atoms) or polyhydric alcohols such as glycols, glycerin, diethylene glycol, petnaerythrite, sorbite, mannite, etc., which, if required, can also be etherified, benzyl benzoate, dioxolanes, glycerin formals, glycol furoles, dimethyl acetamide, lactamides, lactates, ethyl carbonates, silicones (particularly medium-viscosity dimethyl polysiloxanes) and the like.
For example, water or physiologically compatible organic . 30 solvents are suitable for the production of solutions such as, for example, ethanol, 1,2-propylene glycol, polyglycols and their ~;
, ' . .
.
. ., .~ . . - . :
derivatives, dimethyl sulphoxide, fatty alcohols, triglycerides, partial esters of glycerin, para~fin and the like.
For the production of the preparations, Known and conven-tional dissolving intermediaries can be used~ Suitable dissolving intermediaries are polyoxy-ethylated fats, polyoxy-ethylated oleotriglycerides, linolized oleotriglycerides~
Examples of oleotriglycerides are olive oil, peanut oil, castor oil, sesame oil, cottonseed oil, corn oil [see Dr. H.P.
Fiedler "Lexikon der Hilfsstoffe f~r Pharmazie, Kosmetik und - 10 angrenzende Gebiete" (1971), page 191 to 195].
In this connection, "poly-oxylated" means that the sub-stances concerned contain polyoxy-ethylene chains the degree of polymerization of which is usually between 2 and 40 and particu-larly between 10 and 20. Such polyoxy-ethylated substances can be obtained, for example, by reaction of the corresponding glycerides with ethylene oxide (for example, ~0 moles of ethylene oxide per mole of glyceride)~
;,, Moreover, the addition of preservatives, stabilizers, buf~er substances, taste corrigents, antioxidants and complexing agents (for example, ethylenediaminetetraacetic acid) and the like is possible. If required, the pH may be adjusted, for the sta-bilization of the active substance molecule, to approximately
~OS042'7 the usual manner. However, it may~also be carried out by chroma-tography on a silica geI, the silica gel being a synthetically produced, highIy porous a~orphous silica, in the form of hard particles having a granular size of 0.15 to 10 mm. A granular size of 0.15 to 0.30 mm is particularl~ favourable. The water content of this silica can be up to 10~. The specific surface area can he up to 650 sq m per gram. It usually is approximately 4000 sq m per gram. The powder density can be up to 650 g per litre. A powder density of 450 to 500 g per litre is favourable.
Suitable eluants for the silica-gel chromatography of the i organic extract of the roots and rhizomes of ~elleborus species include aliphatic halohydrocarbons, mixtures of aliphatic halo-hydrocarbons with aliphatic alcohols, esters o~ aliphatic acids with aliphatic alcohols, ester-alcohol mixtures, ester-alcohol-water mixtures, benzene, halogen benzenes, alkyl benzenes, alkyl benzene-alcohol mixtures, ester-pyridine mixtures, halohydrocar-bon-pyridine mixtures, halohydrocarbon-alcohol-pyr dine mixtures, ester-pyridine-water mixtures, mixtures of benzene, halogen benzenes and alkyl benzenes with pyridine, aliphatic ketones, ketone-water mixtures, ketone-benzene mixtures, ketone-benzene-acetic acid mixtures, etc. Of course, other mixtures with the ` above components are also possible. The optimum mixing propor-tion of the components must always be determined in a separate preliminary test.
The aliphatic li~uids mentioned above are the usual lower liquid agents used as solvents. In this connection, halogen means fluorine, chlorine or bromine, preferably fluorine or chlorine.
By alkyl benzenes are meant the liquid agents with lower alkyl radicals. Examples are toluene, ethyl henzene, xylenes.
The pre-purified saponin fraction, which is completely free from solvents, is then incubated in water with the enzyme.
.
~ .
~05(~4'~
Toluene can be a~ed (usually in small amounts) to the incubate as a protection against bacterial decomposition.
For the process, commercial cellulase, hemicellulase and ~-glucosidase preparations can be used. Likewise, it is also possible to use preparations of the enzymes produced from fungi, microorganisms (Trichoderma viride), protozoa, bacterial insects, plants, intervertebrates such as snails (edible snails~ and worms by means of the processes known for this purpose [see The Enzymes 1, part 2 (1951) page 729 ff; Ullmann's Enzyklopadie der technis-chen Chemie, 3rd edition (1956) page 391 ff].
The enzymes such as, for example, the cellulase, are obtained by precipitating the culture medium or the aqueous Mycelium extract of the fungi with alcohol, acetone or salt. The crude enzyme can then be further purified, for example, on alu-minium oxide. L
The preparations to be used must be as fresh as possible t without being stored for a lengthy period. Stored preparations must be kept cool and dry. Enzyme preparations or enzyme con-centrates produced from fungi, for example, Aspergillus species such as Aspergillus niger, ~spergillus flavus, Aspergillus oryzae, Aspergillus fumigatus, Aspergillus nidulans are parti-- culary suitable. Preparations or enzyme concentrates produced from Aspergillus niger are particularly favourable.
-; The activity of the enzymes must be determined by pre-liminary tests and the optimum conditions for the reaction and k quantitative relations must also be determined. Enzyme concen-trates which, for example, consist primarily of cellulase, hemi-cellulase or e-glucosidase or of mixtures of such enzymes are particularly favourable. Frequently, such preparations also con- r 30 tain related enzymes such as pectinase (3.2.1.15), amylase (3.2.1.1), acid protoase (3.4.23.6), xylanase (3.2.1.32), cello-biase (3.2.1.21), glucoamylase (3.2.1.3), endopeptidase (3.4.21.16), 5., ' lipase (3.1.1.3), and pectin ~ . . . . . . .
exopolygalacturonase. It is also favourable if the enzvme pre-parations according to the invention also contain enæymes having a macerating effect (for example, ~ectin glvcosidase~. Examples are highIy purified products obtained from Aspergillus species and containing primarily cellulase as hemicellulase as well as pectinase, amylase and acid protease (cellulosin preparations).
The enzymatic reaction isusuallycompleted after two days. If the activity of the enzymes is high, then the reaction is comple-ted earlier, but, if the activity of the enzymes is low, then the treatment is likely to require a much longer period.
The enzyme can be mixed with the substrate either as such or in aqueous solution.
The optimum pH value for the treatment with the enzymes is from approximately 4.5 to 4.7. It is favourable to stir the incubate or to ~eep it in motion in some other manner.
Acid proteases contain cellulosin preparations.
The amount of the enzyme added depends on the activity of the enzyme and on the substrate used. The enzyme can be added, for example, in substantial excess. For example, if a crude saponin fraction which is pre-puri~ied as described hereinbefore is used, then an addition of 5 to 100~ of the enzyme preparation concerned is required depending on the degree of purity, which is determined by thin-layer chromato~raphy. For example, if the drug or the roots and rhizomes of Helleborus species are reacted directly, then an amount from 1 to 10~, relative to the drug or to the roots and rhizomes should be sufficient.
The crude sapogenin obtained after the enzyme treatment can be further processed by means of the usual process describedin -~ ~heliterature~ For example, the process described in Helv. Chim.
~,~
Acta 54, page 1707 (chromatography on SiO2 with a granular size of 0.05 to 0.2 mm) is suitable for this purpose.
_ 5 ~, .
1~)504Z~
Howe~er! a purification (~or example, by chromatography) of the crude sapoqenin with the aid of a silica gel having a grain size of 0~05 to 0,2 mm is particularly favourable. Lower halogenated hydrocarbons such as dichloromethane and chloroform, to which a lower aliphatic alcohol such as methanol or ethanol can be added, are suitable as eluants, the amount of alcohol increasing from 1%.
The sapogenin thus obtained can be recrystallized once more from propanol/water.
10By means of the above process it is possible to isolate the Helleborus sapogenin in substantially larger amounts than by way of the preparatory thin-layer chromatography.
The known Helleborus species, for example, Hellevorus foetidus L., Helleborus multifidus Vis., Helleborus niger L., Helleborus adorus Waldst. et Kit., Helleborus orientalis Lam., Helleboris purpurascens Waldst. et Kit., Helleborus viridis, are suitable starting materials.
Nothir~was previously known concerning the physiological effect of the principal sapogenin from the roots and rhizomes of Helleborus species. It has now been discovered that the princi-pal genin produces from Helleborus saponin has a curative effect on ulcers. Moreover, a muscle-relaxing effect and an effect influencing the central nervous system have also been detected.
The ulcer-protective effect is evident, for example, from the table hereafter:
Daily Dose Ulcer-Protective Effect :
50 mg per kg 58 100 mg per kg 78 200 mg per kg 80 30These tests were carried out on the indomethacin-induced - ulcer of rats using the method of WilheImi which was modified by .. . .. . . : . . :
~504'~7 Jahn and Adrian [see Arzneimittelfor~chung 19 (1969) page 45 ff].
The determination was carried out in the ~ollowing manner:
Albino rats of the strain SIV 50 (S. Ivanovas, Kissle~g/
Allgau) which had an initial weight of 250 to 300 g were kept in wire cages (Ebeco, type 3, double width) in rooms, the temperature of which was kept between 20 and 22C, on a standard diet (Altromin ~). After withholding food for 48 hours, the animals are given 20 mg of indometh~cin in 1.5% tragacanth (l ml of tragacanth solution per 100 g of rat) per kg of rat, applied intragastrally. One hour thereafter the animals are given the test substance orally. The rats continue to stay sober (water ad libitum).
; 24 hours after administration if indomet~a-cin, the animals are killed by means of CO2. The stomachs are resected, opened along the large curvature and rinsed under running water.
The ulcerative changes appear as dark punctiform or ~-striated spots on the mucosa. The evaluation is carried Ollt macroscopically, using the method of MUNCHOW [Arzneimittelfor-schung 4 (1954) page 341].
Therefore, the present invention provides a pharmaceu-tical preparation containing compound I as the active ingredient, if required, mixed with other pharmacologically or pharmaceuti-cally active substances. The medicines can be produce using `; conventional pharmaceutical adjuvant substances.
~ For example, substances which are recommended and/or ,:
listed in the following references from the literature as adju-vant substances for pharmacy, cosmetics and related fields are .; .
;~ suitable as this kind of fillers and adjuvent substances:
Ullmann's Encyklop~die der technischen Chemie, Vol. 4 (1953), - 30 page 1 to 39; Journal of Pharmaceutical Sciences, Vol. 52 ~1963), - page 918 ff, H`.v. Czetsch-Lindenwald, Hilfsstoffe f~r Pharmazie ,~
; - 7 ,~ .
.
... ~ . , 105042~
und angrenzende Gebiete; Pharm. Ind ~ No. 2, 1961, page 72 ff;
Dr. H.P. Fiedler, Lexikon der Hilfsstoffe f~r Pharmazie, Kosmetik and angrenzende Gebiete~ Cantor RG., ~ulendorf i. W~rtt.
1971.
Examples are gelatin, natural sugar such as sucrose or lactose, lecithin, pectin, starch (for example, cornstarch), tylose, talc lycopodium, silica ~for example, colloidal silica), cellulose, cellulose derivatives (for example, cellulose ethers in which the cellulose-hydroxy groups are partially etherified with lower saturated aliphatic alcohols and/or lower saturated aliphatic oxy-alcohols, for example, methyl-oxy-propyl cellulose), stearates, magnesium and calcium salts of fatty acids, containing 12 to 22 carbon atoms, particularly of the saturated ones (for example, stearates), emulsifiers, oils and fats, particularly vegetable oils (for example, peanut oil, caster oil, olive oil, sesame oil, cottonseed oil, corn oll, mono-, di- and trigl~cer-ides of saturated fatty acids C12H24O2 to Cl8H36o2 mixtures), pharmaceutically compatible monohydric or polyhydric alcohols and polyglycols such as polyethylene glycols as well as derivatives thereof, esters o~ aliphatic saturated or unsaturated fatty acids (2 to 22 carbon atoms, particularly 10 to 18 carbon atoms) with monohydric aliphatic alcohols (1 to 20 carbon atoms) or polyhydric alcohols such as glycols, glycerin, diethylene glycol, petnaerythrite, sorbite, mannite, etc., which, if required, can also be etherified, benzyl benzoate, dioxolanes, glycerin formals, glycol furoles, dimethyl acetamide, lactamides, lactates, ethyl carbonates, silicones (particularly medium-viscosity dimethyl polysiloxanes) and the like.
For example, water or physiologically compatible organic . 30 solvents are suitable for the production of solutions such as, for example, ethanol, 1,2-propylene glycol, polyglycols and their ~;
, ' . .
.
. ., .~ . . - . :
derivatives, dimethyl sulphoxide, fatty alcohols, triglycerides, partial esters of glycerin, para~fin and the like.
For the production of the preparations, Known and conven-tional dissolving intermediaries can be used~ Suitable dissolving intermediaries are polyoxy-ethylated fats, polyoxy-ethylated oleotriglycerides, linolized oleotriglycerides~
Examples of oleotriglycerides are olive oil, peanut oil, castor oil, sesame oil, cottonseed oil, corn oil [see Dr. H.P.
Fiedler "Lexikon der Hilfsstoffe f~r Pharmazie, Kosmetik und - 10 angrenzende Gebiete" (1971), page 191 to 195].
In this connection, "poly-oxylated" means that the sub-stances concerned contain polyoxy-ethylene chains the degree of polymerization of which is usually between 2 and 40 and particu-larly between 10 and 20. Such polyoxy-ethylated substances can be obtained, for example, by reaction of the corresponding glycerides with ethylene oxide (for example, ~0 moles of ethylene oxide per mole of glyceride)~
;,, Moreover, the addition of preservatives, stabilizers, buf~er substances, taste corrigents, antioxidants and complexing agents (for example, ethylenediaminetetraacetic acid) and the like is possible. If required, the pH may be adjusted, for the sta-bilization of the active substance molecule, to approximately
3 to 7 with physiologically compatible acids or buffers. In general, a pH as neutral as possible to weakly acid (to pH 5) is preferred.
~ or example, sodium metabisulphite, ascorbic acid, gallic acid, gallic alkyl ester, butyl hydroxy anisole, nor-dihydro guaiaric acid, tocopherols as ~ell as ~ocopherols + synergists , .:,.
(substances which bind heavy metals by complexing action, for example, lecithin, ascorbic acid, phosphoric acid) are used as antioxidants~ The addition of the synergists substantially ,.:' .
, - g _ :
~OS04~7 ~ ~
increases the antioxygenating action of the tocopherols.
Suitable preservatives include sorbic acid, p-hydroxy benzoic esters (for example, lo~er alk~1 ester~, benzoic acid~
sodium benzoate, trichloro isobutyl alcohol, phenol, cresol, benzethonium chloride and formalin derivatives.
The pharmacological and galenic handling of the compounds according to the invention is carried out according to conven-tional standard methods. For example, the active ingredient and adjuvant substances or fillers are properly mixed by stirring or homogenizing (~or example, by means of colloid mills, ~all mills), usuall~ at a temperature of 20 to 80C, prefera~ly from 20 to 50C.
The medicines can be applied orally, parenterally, rect-ally, vaginally, perlingually or locally. The addition of other active medical substances is also possible.
The compound shows a good antiulcerogenic effect on the indomethacin ulcer of rats and on the histamine-induced ulcer of guinnea pigs.The antiulcerogenic effect is comparable to the effect of the known drug biogastrone.
The testing was carried out on histamine-induced ulcers of guineapigs by means of the method of Eagleton and Watt (Peptic Ulcer, Pfeiffer, Munksgaard, Copenhagen, printed in Denmar~ of Aarhus Stiftsbogtrykkerie A/S ~rhus ISBN 8716003225). In this test, the test substances are given orally to male guinae pigs ' (weighing 300 to 400 g each) after withholding food for 24 hours and drinking water for 12 hours. One hour after administering i the substance, the animals are injected with histamine diphos-;~ phate (dose: 5 mg/kg). The histamine diphosphate is applied intraperitoneally. Three hours after the histamine injection the animals are killed by means of CO2. The stomachs are resected and cut open along the small curvature and inverted over the , - 10 ~
testing finger, The ulcers axe evaluated according to M~nchow (Arzneimittelforschung 4, 341, 1954~
The computed control index must be o~ the order of 5 to 15.
The lowest effective dose in the animal test mentioned hereinbefore is, for example, 50 mg per kg orally. For example, 50 to 500 mg per kg orally is suitable as a general dose range for the above effect (animal test as above).
The compound according to the invention is indicated for Ulcus ventriculi, Ulcus duodeni, gastritis and duodenitis.
The pharmaceutical preparations usually contain between 1 and 50% of the active component according to the invention.
They can be given, for example, in the form of tablets, capsules, dragees or in a liquid form. Oily or alcoholic solu-tions and emulsions are suitable as li~uid forms of application.
Tablets containing between 50 and 500 mg of the active substance or solutions containing between 0.5 and 10% thereof are preferred forms of application. For oral medicines, the single dose of the active component according to the invention can be between 50 and 500 mg. For example, three times daily 1 to 5 tablets containing from 50 to 500 mg of active substance can be recommended.
In mice, the acute toxicity of the compound according to the invention [expressed by the LD 50 mg/kg; method according to Miller and Tainter; Proc. Soc. Exper. Biol. a. Med. 57 (1944) Z61] is above 6000 mg per kg for oral application. ~-The invention will now be described in greater detail by means of the following examples, which describe pharmaceutical preparations containing the principal sapogenir. of roots and rhizomes of the Helleborus species, and a process for extracting such sapogenin.
:
;
~5V4Z7 Example 1 (Tablets) _ 50 g of the principal sapogenin of Helleborus are mixed with 50 g of highIy dispersed silica, 10 g of cornstarch and 120 g of lactose. The powder is granulated with a solution of 5 g of methyl-oxy-propyl cellulose in approxi~atel~ 160 ml of a 30% ethanol. The dried granulate is mixed with 21 g of corn-starch, 18 g of talc, 125 g of microcrystalline cellulase and 1 g of magnesium stearate, whereupon the mixture is pelletized in a known manner. One tablet, which weighs 400 mg, contains 50 mg of sapogenin. `
Example 2 (Gelatin Capsules) 500 g of dimethyl pol~siloxane having a viscosity of 360 to 380 cp at 20C (silicone oil AK 350) are triturated with 20 g of highly dispersed silica. The triturate is processed on a water bath with 130 g of isopropyl myristinate, 150 g of a melted mixture of mono-, di- and tri-ylycerides of the saturated ' 12 242 C18H36O2, a commercial product known as hard fat (Deutsches ~rzneibuch 7th edition) and 200 g of prin-cipal sapogenin of ~elleborus. A homo~eneous paste is thus obtained.
The paste is put into gelatin capsules in single doses . . .
of 500 mg. One gelatin capsule contains 100 mg of sapogenin.
Example 3 (Coated Tablets, Dragees) Tablets or dragee cores, which are produced corresponding to Example 1, can be provided in a known manner with a film coat-ing, which is soluble in the stomach or in the small intestine, ~` and which consists of suitable cellulose derivatives or other polymeric film forming agents and suitable additives such as `~ plasticizers, dyes, fla~oring substances, etc., or with a suitable dragee coating.
, ,~
~, .
1051~4Z7 Example 4 (Alcoholic Solution) 7.5 g o~ principal sapogenin of Helleborus are dissolved with 0.5 g of cherry aroma in a 96% ethanol and the solution is diluted with ethanol to lO0 ml. For the peroral application, 1 ml of the solution is mixed with approximately 50 to 100 ml of liquid. l ml of solution contains 75 mg of sapogenin.
Example 5 100 kg of roots and rhizomes of Helleborus viridis are comminuted, degreased with petroleum ether and then thoroughly extracted with an 80% aqueous ethanol. The residue of the ethanol extract is taken up in water, and the solution is shaken with chloroform/ethanol (2:1). The chloroform-soluble components are chromatographed with chloroform/ethanol (9:1) on a silica-gel column, and the fractions are examined by means of thin-layer chromatography:
adsorbent; silica gel for thin-la~er chromatography (silica gel G of the firm ~f Merck sRD) solvent: chloroform/methanol/water (35:25:10) detection: anise aldehyde/sulphuric acid/acetic acid tl:l:100).
The fraction containin~ primarily a substance which can be colored brown, has an average Rf value (approximately O.S0) and is localized between des-glucohellebrin and helle~rin in the chromatogram.
The residue of the combined fractions ~2621 g) is diss-olved in 2.4 litres of methanol and 2.4 litres of warer with reflux. Upon adding 24 litres of hot water, 4.8 litres of sol-vent are distilled from the solution.
Upon cooling, the solution is treated with 240 g of a ; ~ commercial cellulase (R~hm and Haas) and lO0 ml of toluene, and stored at 40C while shaking occasionally. The reaction is completed after approximately 48 hours. The precipitate is .
. ~
_ 13 ~504'~7 filtered with suction, washed with hot water and dried (1100 g).
The dried precipitate is dissolved in methanol and ; dichloromethane, and the solution is chromatographed with dichIoromethane~methanol (increasing methanol concentration) on silica gel for the column chromatography with a granular size of 0.2 to 0.5 mm.
The sapogenin fraction (736 g) is dissolved in 5 litres of propanol-(l), and the solution is treated with 30 litres of water. After 24 hours, the separated crystals are washed with propanol/water and dried.
m.p. = 238 to 240C
; [a] D = - 86.93C (c=l.l; pyridine) ,. .
.
, . "
., ' .
. :
.~
':
t`' ~ 30 " .
' "' ~ . . .
~ or example, sodium metabisulphite, ascorbic acid, gallic acid, gallic alkyl ester, butyl hydroxy anisole, nor-dihydro guaiaric acid, tocopherols as ~ell as ~ocopherols + synergists , .:,.
(substances which bind heavy metals by complexing action, for example, lecithin, ascorbic acid, phosphoric acid) are used as antioxidants~ The addition of the synergists substantially ,.:' .
, - g _ :
~OS04~7 ~ ~
increases the antioxygenating action of the tocopherols.
Suitable preservatives include sorbic acid, p-hydroxy benzoic esters (for example, lo~er alk~1 ester~, benzoic acid~
sodium benzoate, trichloro isobutyl alcohol, phenol, cresol, benzethonium chloride and formalin derivatives.
The pharmacological and galenic handling of the compounds according to the invention is carried out according to conven-tional standard methods. For example, the active ingredient and adjuvant substances or fillers are properly mixed by stirring or homogenizing (~or example, by means of colloid mills, ~all mills), usuall~ at a temperature of 20 to 80C, prefera~ly from 20 to 50C.
The medicines can be applied orally, parenterally, rect-ally, vaginally, perlingually or locally. The addition of other active medical substances is also possible.
The compound shows a good antiulcerogenic effect on the indomethacin ulcer of rats and on the histamine-induced ulcer of guinnea pigs.The antiulcerogenic effect is comparable to the effect of the known drug biogastrone.
The testing was carried out on histamine-induced ulcers of guineapigs by means of the method of Eagleton and Watt (Peptic Ulcer, Pfeiffer, Munksgaard, Copenhagen, printed in Denmar~ of Aarhus Stiftsbogtrykkerie A/S ~rhus ISBN 8716003225). In this test, the test substances are given orally to male guinae pigs ' (weighing 300 to 400 g each) after withholding food for 24 hours and drinking water for 12 hours. One hour after administering i the substance, the animals are injected with histamine diphos-;~ phate (dose: 5 mg/kg). The histamine diphosphate is applied intraperitoneally. Three hours after the histamine injection the animals are killed by means of CO2. The stomachs are resected and cut open along the small curvature and inverted over the , - 10 ~
testing finger, The ulcers axe evaluated according to M~nchow (Arzneimittelforschung 4, 341, 1954~
The computed control index must be o~ the order of 5 to 15.
The lowest effective dose in the animal test mentioned hereinbefore is, for example, 50 mg per kg orally. For example, 50 to 500 mg per kg orally is suitable as a general dose range for the above effect (animal test as above).
The compound according to the invention is indicated for Ulcus ventriculi, Ulcus duodeni, gastritis and duodenitis.
The pharmaceutical preparations usually contain between 1 and 50% of the active component according to the invention.
They can be given, for example, in the form of tablets, capsules, dragees or in a liquid form. Oily or alcoholic solu-tions and emulsions are suitable as li~uid forms of application.
Tablets containing between 50 and 500 mg of the active substance or solutions containing between 0.5 and 10% thereof are preferred forms of application. For oral medicines, the single dose of the active component according to the invention can be between 50 and 500 mg. For example, three times daily 1 to 5 tablets containing from 50 to 500 mg of active substance can be recommended.
In mice, the acute toxicity of the compound according to the invention [expressed by the LD 50 mg/kg; method according to Miller and Tainter; Proc. Soc. Exper. Biol. a. Med. 57 (1944) Z61] is above 6000 mg per kg for oral application. ~-The invention will now be described in greater detail by means of the following examples, which describe pharmaceutical preparations containing the principal sapogenir. of roots and rhizomes of the Helleborus species, and a process for extracting such sapogenin.
:
;
~5V4Z7 Example 1 (Tablets) _ 50 g of the principal sapogenin of Helleborus are mixed with 50 g of highIy dispersed silica, 10 g of cornstarch and 120 g of lactose. The powder is granulated with a solution of 5 g of methyl-oxy-propyl cellulose in approxi~atel~ 160 ml of a 30% ethanol. The dried granulate is mixed with 21 g of corn-starch, 18 g of talc, 125 g of microcrystalline cellulase and 1 g of magnesium stearate, whereupon the mixture is pelletized in a known manner. One tablet, which weighs 400 mg, contains 50 mg of sapogenin. `
Example 2 (Gelatin Capsules) 500 g of dimethyl pol~siloxane having a viscosity of 360 to 380 cp at 20C (silicone oil AK 350) are triturated with 20 g of highly dispersed silica. The triturate is processed on a water bath with 130 g of isopropyl myristinate, 150 g of a melted mixture of mono-, di- and tri-ylycerides of the saturated ' 12 242 C18H36O2, a commercial product known as hard fat (Deutsches ~rzneibuch 7th edition) and 200 g of prin-cipal sapogenin of ~elleborus. A homo~eneous paste is thus obtained.
The paste is put into gelatin capsules in single doses . . .
of 500 mg. One gelatin capsule contains 100 mg of sapogenin.
Example 3 (Coated Tablets, Dragees) Tablets or dragee cores, which are produced corresponding to Example 1, can be provided in a known manner with a film coat-ing, which is soluble in the stomach or in the small intestine, ~` and which consists of suitable cellulose derivatives or other polymeric film forming agents and suitable additives such as `~ plasticizers, dyes, fla~oring substances, etc., or with a suitable dragee coating.
, ,~
~, .
1051~4Z7 Example 4 (Alcoholic Solution) 7.5 g o~ principal sapogenin of Helleborus are dissolved with 0.5 g of cherry aroma in a 96% ethanol and the solution is diluted with ethanol to lO0 ml. For the peroral application, 1 ml of the solution is mixed with approximately 50 to 100 ml of liquid. l ml of solution contains 75 mg of sapogenin.
Example 5 100 kg of roots and rhizomes of Helleborus viridis are comminuted, degreased with petroleum ether and then thoroughly extracted with an 80% aqueous ethanol. The residue of the ethanol extract is taken up in water, and the solution is shaken with chloroform/ethanol (2:1). The chloroform-soluble components are chromatographed with chloroform/ethanol (9:1) on a silica-gel column, and the fractions are examined by means of thin-layer chromatography:
adsorbent; silica gel for thin-la~er chromatography (silica gel G of the firm ~f Merck sRD) solvent: chloroform/methanol/water (35:25:10) detection: anise aldehyde/sulphuric acid/acetic acid tl:l:100).
The fraction containin~ primarily a substance which can be colored brown, has an average Rf value (approximately O.S0) and is localized between des-glucohellebrin and helle~rin in the chromatogram.
The residue of the combined fractions ~2621 g) is diss-olved in 2.4 litres of methanol and 2.4 litres of warer with reflux. Upon adding 24 litres of hot water, 4.8 litres of sol-vent are distilled from the solution.
Upon cooling, the solution is treated with 240 g of a ; ~ commercial cellulase (R~hm and Haas) and lO0 ml of toluene, and stored at 40C while shaking occasionally. The reaction is completed after approximately 48 hours. The precipitate is .
. ~
_ 13 ~504'~7 filtered with suction, washed with hot water and dried (1100 g).
The dried precipitate is dissolved in methanol and ; dichloromethane, and the solution is chromatographed with dichIoromethane~methanol (increasing methanol concentration) on silica gel for the column chromatography with a granular size of 0.2 to 0.5 mm.
The sapogenin fraction (736 g) is dissolved in 5 litres of propanol-(l), and the solution is treated with 30 litres of water. After 24 hours, the separated crystals are washed with propanol/water and dried.
m.p. = 238 to 240C
; [a] D = - 86.93C (c=l.l; pyridine) ,. .
.
, . "
., ' .
. :
.~
':
t`' ~ 30 " .
' "' ~ . . .
Claims (10)
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A pharmaceutical preparation comprising 50 to 500 mg of the principal sapogenin of the roots and rhizomes of Helleborus species, and a pharmaceutically acceptable carrier.
2. A preparation according to claim 1, wherein said carrier is selected from the group consisting of gelatin, sucrose, lactose, lecithin, pectin, cornstarch, tylose, talc lycopodium, silica, cellulose, cellulose ethers in which the cellulose-hydroxy groups are partially etherified with at least one of lower saturated aliphatic alcohols and lower saturated aliphatic-oxy-alcohols, stearates, magnesium and calcium salts of fatty acids, containing 12 to 22 carbon atoms, emulsifiers, peanut oil, castor oil, olive oil, sesame oil, cottonseed oil, corn oil, mono-, di- and triglycerides of saturated fatty acids C12H24O2 to C18H36O2 and mixtures thereof), monohydric or polyhydric alcohols and polyglycols and derivatives thereof, esters of aliphatic saturated or unsaturated fatty acids containing 2 to 22 carbon atoms with monohydric aliphatic alcohols containing 1 to 20 carbon atoms or polyhydric alcohols, benzyl benzoate, dioxolanes, glycerin formals, glycol furoles, dimethyl acetamide, lactamides, lactates, ethyl carbonates, silicones, ethanol, 1,2-propylene glycol, polyglycols and their derivatives, dimethyl sulphoxide, fatty alcohols, triglycerides, partial esters of glycerin and parrafin.
3. A preparation according to claim 1, including at least one of a preservative, stabilizer, buffer, taste corrigent, antioxidant and complexing agent.
4. A preparation according to claim 1 or 2, including, as a preservative, sorbic acid, a p-hydroxy benzoic ester, benzoic acid, sodium benzoate, trichloro isobutyl alcohol, phenol, cresol, benzethonium chloride or a formalin derivative.
5. A preparation according to claim 1, in tablet form, including the principal sapogenin of the roots and rhizomes of the Holleborus species, silica, cornstarch, lactose, methyl-oxy-propyl cellulose, talc, microcrystalline cellulase and magnesium stearate.
6. A preparation according to claim 1, in paste form for capsules, including dimethyl polysiloxane, silica, isopropyl myristinate, a mixture of mono-, di- and triglycerides of the saturated fatty acids C12H24O2-C18H36O2 and the principal sapo-genin of the roots and rhizomes of the Helleborus species.
7. A preparation according to claim 1, in solution form, including the principal sapogenin of the roots and rhizomes of the Helleborus species, cherry aroma and ethanol.
8. A process for extracting the principal genin con-tained in the roots and rhizomes of Helleborus species comprising treating at least one of the roots and the rhizomes of Helleborus species with an enzyme selected from the group consis-ting of cellulase, hemicellulase and .beta.-glucosidase.
9. A process according to claim 8, wherein the reaction mixture obtained after the treatment with the enzyme is purified using silica gel.
10. A process according to claim 9, wherein the silica gel is granular with a particle size 0.2 to 0.5 mm.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE2416979A DE2416979C3 (en) | 1974-04-08 | 1974-04-08 | Process for obtaining the main sapogenin contained in the roots and rhizomes of Helleborus species |
| DE2416978A DE2416978A1 (en) | 1974-04-08 | 1974-04-08 | MEDICINAL PRODUCTS WITH THE MAIN APOGNANT OF THE HELLEBORUS SPECIES AS THE ACTIVE SUBSTANCE |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1050427A true CA1050427A (en) | 1979-03-13 |
Family
ID=25766942
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA224,029A Expired CA1050427A (en) | 1974-04-08 | 1975-04-08 | Pharmaceutical preparation containing the principal helleborus sapogenin |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA1050427A (en) |
-
1975
- 1975-04-08 CA CA224,029A patent/CA1050427A/en not_active Expired
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