Note: Descriptions are shown in the official language in which they were submitted.
The invention relates to medicaments with prostaglandin-
synthetase (PGS) inhibiting activity.
Prostaglandins (PGs) are widely present in all mammalian
organisms. For some years scientific research has been concentrated
in intensive efforts for extracting and acknowledging the bio-
logical activity of prostaglandins. Arcording to present
knowledge multiple PGS or precursors exist which vary somewhat
in structure and of which the biological importances are wide
spread occurrence,high activities and differences in metabolic
actionO These different actions are based on the fact that the
intracellular PG synthesis can be induced by irritation or damage
of cellular membranes during which in th~ first phase phospho-
lipases release PG-precursors from membrane lipids, or that on
the other hand several hormones e.g. bradykinines, acetylchlorine
or histamine increase the synthesis and release of PGs and that
moreover PGs not only stimulate the adenyl-cyclase-system but also
the guanyl--cyclase-system and therefore can cause an increase of
intracellular APM- and GPM- concentrations.
It had already been known that the PG e~ects vary
depending on PG types used and tested organs, e.g. the
adenyl-cyclase is stimulated in endocrine organs by PGEl and
PGE2 but inhibited in fat tissue. This fact explains why PGs
are able to increase or decrease the APM level in a target organ
and display an adrenalin- and glucagon antagonistic effect in fat
tissue. With smooth muscles PGs in part effect contractions
e.g~ in uterus or intestine or dilatation e.g. in blood vessels.
Cl
PGs E2 and A2 increase secretion of sodium and potassium in the
~idneys. Furthermore, it had been established already that an
increase of PGs E2 and F2 level in the tissue can initiate and
maintain inflammatory reactions.
The multiple metabolic effects of PGs are basis for
several therapeutic uses. Thus, PGs are used in the treatment
of asthma and circulatory diseases because PGs of the E-type
have vessel-dilatating activity. On the other hand, PGs induce
labor and initiate parturition so that it is possible to use them
for inducing abortion.
It was not known until recently that the activity of some
medicaments with analgetic and anti-inflammatory action which had
already been used for decades is based on an inhibition of the
prostaglandin synthetases. This applies for e.g. acetylsalicylic
acid, indomethacine or ibuprofen. The strong inhibitory action
of these compounds explains the activity against inflammations on
one hand and on the other hand the presence of several side e~fects
of which only the induction of stomach hemorrhages is referred to.
The biosynthesis of the PGs starts from membrane
phospholipides which are converted into arachidonic acid and are
reacted into endoperoxide - PGs by oxygen radicals. By further
reactions the relatively stable PGs, thromboxanes and the
relatively unstable prosta-cyclin are formed from the endoperoxide -
PGs.
The synthesis of PGE2 and PGF2~ from arachidonic acid
can be shown in a shortened way in the following formula:
f~ I
- ~ , ' :
' ' ' ' '
\ COOH
OH
COOH
~ ",.
OH
~1 ~ .
COQH
OH
OH
The biological residence time of PGs and their precursors
is only miniscule. The decomposition starts from oxidation at
C-5 and continues via the ~-oxidation of fatty acids.
It had already been established that certain chemical
compounds are strong PGS inhibitors. These compounds e.g.
indometacine or acetylsalicylic had been found to be PGE2 -
synthetase inhibitors and are used correspondingly in the
treatment of rheumatic and arthritic and similar conditions for
instance. The strong inhibitor action which is not necessarily
restricted to PGE2 - synthetase leads to unwanted side effects
~J~
'~
based on this action e.g. initiation of bleeding in the stomach
and intestine, other scattered bleedings,occUrrenCe of allergies
or possibilities of influencing gravidity.
Therefore, it is the object of the invention to develop
a new medicament with an activity as PGS-inhibitor which does
not possess the known disadvantages.
The invention accordingly provides a pharmaceutical
preparation in unit dosage form, active as a PGS inhibitor,
comprising from about 0.01 to 10 mg. per dosage unit of a glycoside
10 selected from sterol glycosides and esters thereof and spiroketal-
steroid glycosides and esters thereof, in a pharmaceutically
acceptable carrier.
Surprisingly and completely unexpected it has now been
established that sterolglycosides, spiroketal-steroidglycosides
and esters thereof are active inhibitors for PGE2 - and PGF2~
synthetases without producing side effects which normally occur
in influencing the PG level. Sterolins are compounds occurring
frequently though only in small amounts in nature~in plants
and microorganisms. Sterolins are chemically glycosides of
phyto-sterols including cholesterol and sterol type tetracyclic
triterpenes as for example lanosterol and cycloartenol. Some
of these compounds occur in various plants in relati~ely larger
amounts, e.g., campesterol and stigmasterol and particularly
sitosterol. The phytosterols correspond to the following
general formula:
, j j . .
:
,
" , '
' ~ :
R4
., .
HO ,' \
R4 R2
in which R , R2 and R3 are hydrogen atoms or methyl groups and
in which R4 can be a hydrogen atom or a methyl, ethyl, methylene
or ethylidene groupO Furthermore, there can be present double
bonds at various places of the basic structure. This is true
also for the side chain.
Phytosterols are present in most plants partially as
steroglycosides i.e. therefore as sterolins and sometimes as
their esters. The most common sterolins in nature are
monoglycosides though a few diglycosides had been described as
well. Besides the predominantly occurring D-glucose which is
joined to the s-terol by the 3-~-hydroxy group, mostly by
equatorial or ~-glycoside bonds, mannose, galactose, arabinose
and xylose has been found in the naturally occurring compounds.
As far as esters which occur in nature, these have been identified
as esters of monobasic monocarboxylic acids.
C,
Spiroketal-steroidglycosides are those steroidsaponins
which carry at the steroid basic structure - which is the aglycone -
at carbon atoms 16 and 17 a spiroketal group. The aglycones can
be classified in being 5-ene-steroid sapogenins or 5~-steroid
sapogenins. 5-ene-compounds are for example diosgenin, yamogenin,
botogenin and correlogin while representatives of 5-~-compounds
are for example tigogenin, neotigonin, hecogenin and sisalagenin.
In nature the aglycones of the saponins exist as glyco-
sides which carry in general 3 or more monosaccharide units in
the sugar moity.
sased on this fact these compounds which are relatively
rich in sugar dissolve rather readily in water and frequently
give rise to a soap like froth.
The spiroketal-steroidglycosides correspond to the
following general formula
, ~\0~
Z--O
~,
.
.:, ~ .
where in position 5 a double bond or an ~-hydrogen atom can be
present and where Z is a mono- or disaccharide, especially glucose,
which can be esterified.
The sterolins and spiroketal-steroidglycosides used
according to the invention and their esters can be dispensed as
extracts o-f plant material, enriched extracts or compounds
produced synthetically.
The synthesis can be carried out in a manner known per se
as for example the known Kornigs-Knorr-synthesis for producing
glycosides using the corresponding aglyconest a C-l brominated
sugar acetate and silver oxide or silver carbonate.
In case of treatment with sterolins or their esters it is
to be noted that these compounds are extremely insoluble in water.
Therefore, for administration, sterolins must have an appropriately
small particle size which enables resorption. It is absolutely
necessary that the sterolins used in the invention are produced
and/or prepared and/or incorporated in pharmaceutical preparations
in such a manner that liquid or solid solutions, emulsions or solid
dispersions are formed which can be made in a known way by
adsorption, absorption or grinding with or without auxiliary
material. All these methods lead to a reduction of particle size
and crystallinity so that the compounds no longer exist as
crystalline micro particles but as miniscule amorphous mono- or
multilayer aggregates.
The compounds o-f the invention generally are used with a
particle size of about 0.1 mm and preferably 0.06 mm and smaller.
The same applies to the spiroketal-steroidglycosides
`' ~ 1
s ~
not-withstanding their better solubility in water. Ihese compounds
too ar~ used with a particle size of 0.1 mm and preferably 0.06 mm
or smaller~
The compounds of the invention are administered in daily
doses of about 0.03 to 10 mg. The usual curati~e or protective
dosage is about 0.1 to 0.45 mg daily. The doses can be given in 3
single doses or in a single dose with slow release of active
material.
According to present knowledge the compounds are suited
for treatment of diseases in which a reduction of the PGE2 or PGF2
level is necessary. Among these diseases are the following ones:
1.) Ulcers, especially of stomach and intestine,
2.) endocrine disregulations,
3.) urogenital diseases especially benign prostata
hypertrophy and accompanying diseases,
4.) heart illnesses and blood pressure disturbances,
5.) edematose conditions,
6.) blood vessel diseases, thrombosis varicose veins
and hemorrhoids,
7.3 dermatitis and reactions based on excess of
histamine,
8.) inflammatory reactions,
9.) arthritic and rheumatoid diseases, ;~
10.) allergies including asthma.
The compounds of the invention can be used in corresponding
manner to treat animal diseases. The dosage used in animal
diseases can be calculated in a known manner by taking 75 kg
as the average human weight.
--8--
'' ' ~'
The compounds can be prepared in usual way known to the
expert as pharmaceutical compositions, e.g. powders, pills,
tablets, capsules, dragées, emulsions, solutions, injection - or
infusion solutions, ointments or creams.
The invention will be further explained in connection
with the examples.
Example 1
Production of Sitosterol-~-D-Glucoside.
A mixture of 41,4 grams of sitosterol and 55,2 grams of
silver carbonate in toluene were distilled with stirring until the
distillate passed over free of water. Then there was added to the
stirred, boiling mixture dropwise a solution of 82,2 grams of
acetobromoglucose in 100 ml of toluene. The toluene was
continuously further distilled so that the total amount of water
formed in the reaction was removed azeotropically. The reaction
vessel was protected from light during this time. In case it
is necessary, the volume of the reaction mixture is held constant
by addition of dry toluene. After the addition of the bromo-
acetoglucose solution it was maintained at the boiling point until
the distillate was water free. Subsequently the reaction mixture
was filtered and the residue washed with fresh hot toluene. The
united filtrate and washing liquids were then evaporated under
reduced pressure. The residue was recrystallized from ethanol
or hexane. The yield of sitosterol glucoside tetraacetate was
22~4 grams, corresponding to 30%.
A solution of 1 gram of sodium in 100 ml of ethanol
:
under stirring was added quickly to a solution of 10 grams
sitosterol glucoside tetraacetate in 600 ml ethanol at a
temperature of 45C. The mixture was stirred for one hour before
2 liters of water were added and the mixture was then stirred for
another hour. The precipitated sitosterol glucoside was filtered
off and washed until neutral with water prior to drying it for
12 hours in a vacuum. The yield was 6,0 grams, corresponding to
95%.
By selection of suitable starting compounds all the
remaining mentioned sterolins also can be produced according to
the above given process.
_xample 2
S~nthesis of diosgenin-3-~-D-glucoside.
A well stirred mixture of 55,2 g silver carbonate and a
solution of 41,4 g diosgenin in 600 ml toluene was distilled unti~
the collecting distillate was free of water. A solution of 82,2
g bromoacetylglucose in 100 ml toluene was then added dropwise to
the stirred mixture, boiling continuing all the time with
distillation in order to remove the water formed during the
glucosidation reaction. At this stage the reaction vessel is
protected from light and the volume of the reaction mixture is
kept constant at about 500 ml by adding extra dry toluene whenever
necessary. After the acetobromoglucose addition has been completed
distillation is continued until no further water separates from the
condensate. The reaction mixture is then filtered hot and the
residue is washed with fresh, hot toluene~ The combined filtrate
and wash solutions are evaporated to dryness under vacuum and the
reSldUe lS
--10--
,
.
crystallised from ethanol or hexane. The yield of
diosgenin-3-~-D~$1ucoside tetraacetate l~a~ 25,5 g
or 3~.3%
To a stirred solution of lo g diosgenîn tetraacetate
in 600 ml ethanol At ~5 C i9 rapidly added 15 ml
of an ethanolic solution of 1 g sodium in loo ml
abs. ethanol~ The stirred mixture is allowed $o
react for 1 hour bsfore 2 1 water is added and this
mixture i~ then stirred for a further hour. The
precipitated diosgenin glucoside i~ then filtered
off and washed to neutrality ~ith water before it is
dried under vacu~ at looC for 12 hours. The yield
was 7 g or 90%.
Similarly the further spiroketal-steroid-glycGsides
can be prepared.
. .
~xample 3
The_preparation of .pharmaceutical products.
~a) The preparation of lact:ose-corn starch powders
incorpor~ting diosgenirL-~-D-glucoside:
A boiling ho* solut~8.n of 15 g di~sgenin glucoside
in 2,25 1 chloroforn~ and 750 ml ethanol i~ mixed
with 1 ~g lactose powder of a particle si~e no*
exceedin~ o,15 mm. The resulting slurry is dried
~ith constant stirring and the impregnated lactose
is ~educed to its original particle ~ize before it
is mixed ~ith 9 kg corn starch and 50 g maglles.ium
stearate. Capæules are rcadil~ filled with this
mixture. ~hus a capsule containing l~o mg of the
~ixt-lre l~ill ca.rry approximately o,15 mg diosgenin~
D-glucoæi~le, lo ms of lactose, 90 mg of corll starch
a~ well as o,5 mg of magneæium stearate.
- 12 -
(b3 The preparation of lacto~e granu~¢es containing
diosgenln-~3-D-glucoside:
A boi].ing hot solution of 5 g diosgenin glucoside
in 5 1 ethanol is mixed with 3,32 k~ of a particle
~ize not exceeding o,15 mm. The agi.tated slurry
i5 dried and the impregnated lactose i5 then r~duced
to the original particle size before it is converted
into granules of o,7-1-2 mm particle sizes. This
granula~ed product i~ al~o suitable ~or filling
into capsules, where for example a capsule contain~ng
loo mg granulate carries o,15 mg diogenin-~-D-glucoside.
Produc*s as de3cribed under a) and b) can also be
prepared using the fol~owing: ~
'
i) Glyeo~ide~ of the mentioned ~ydrox~spiroke~al-~teroids
and especially the ~-D-gluco.sides of ~`igogenin and
Hecosenin,
il) gluco~e, ascorbic acid or t~lc as carrier~ for the
glycosides or any other inert and pharmace~.ically
acceptable carrier,
iii) the contents of actiYe glycoside in each cap~ule can
be adju~ted to any ~alue between o,ol and more,~
i~) the auxilliary sub~tances described in a) and b)
can be altered to accepted pharmaceutical prace~ces,
. v) at each stage of the production process described
under a3 and b3 other pharmaceutically actiYe
~ubstan^es can be added and incorporated into the
iinal product.
, .
.
.
. .
- 13 ~ 3~ ~
(c) The preparation of tablets containing diosgenin
B-D-glucoside:
A slurry, prepared by mixing 9co g lactose with a
hot solutioll of 1,25 g diosgenin glucoside in 1 1
chloroform, i9 dried at room temperature and under
a vacuum with constant agitation. The powdered
product is then thoroughly mixed with 2,1 kg
potato starchO The impregnated lactose-starGh mixt~re
i~ granulated by treating it with 2,5 1 of an
aqueous solution containing 250 g geiatine and
5 g glycerine. The granulate, dried under reduced
pressure at room temperature is converted by kno~n
methods into ~oo mg *ablets~ Each table* then
con~ainq o,15 mg diosgenin ~-D-glucoside, llo,56 mg
lac-tose, 257~97 mg potato starch, 30,31 mg gelatine
~nd o,61 mg glycerine.
(d) The preparation of hecog~enin ~-D_~lucoside con~aining
dragee~.
A slurry prepared by mixing 1850 S lactose, 300 g
~ucrose and a hot ~olution of 450 mg hecogenin gluooside
in 2 1 chloroform is dricd under a vacuum at 30C.
The powdered product is granulated by knol~n me*hods
by adding 1,6 1 of an aqueous solution of 40 g
gelatineO The granulate, dried under reduced pressure
at ~C and thoroughly l~ixed with lo g magnesium
; stearate, is con~erted into 3000 pressed Xernels
which are finally coated to produce drasees.
Every dragée th0n contains o,l5 mg hecogenin
~-D-$1ucoside, 616,67 mg lactose, loo,oo mg sucrose,
13,33 mg gelatine and 3,33 mg magneslwn stearate.
(e) Preparation o~ an ointment containing hecogenin
~-D glucoside:
~ ~ .
'
,
To a mixture of 1 g hecogenin glucoside worked into
emulsifying 90 g cetyl alcohol is added 105 g viscous
paraffin and 105 g white Vaseline* before melting the
complete mi~ture on a 60C waterbath. Into this melt is
stirred 699 g water in small portions at a time. The
final mixture is stirred until cold to provide an ointment
containing 0,1% glucoside.
(f) Preparation of a cream containing tigogenin ~-D-glucoside:
500 g woolwax alcohol into which 1 g tigogenin glucoside
has been incorporated is heated to about 50C on a water
bath. To the resulting melt 499 g water is added in small
portions with constant stirring. The final cream is
stirred until it reaches room temperature; during this
process sufficient water is added to replace evaporation
losses. This cream contains 0,1% glucoside.
Similarly the other compounds mentioned may be incorporated
into pharmaceutical preparations.
E~ample 4
Production of pharmaceutically acceptable solutions
(a) Solution having a content of semisynthetic Soya Sterol-~ D-
Glucoside
There was added to a boiling solution of 600 mg of semi-
synthetic soya sterol-~-D-~lucoside in 6 liters of absolute ethanol
a solution of 10 grams o~ polyvinyl pyrrolidone in 4 liters of
distilled water having a temperature of 65C. The cooled 60%
ethanolic solution was filled with 250 ml flasks. The patients were
directed
.
*Trade mark
-14-
~ Cl
.
.
,
.~ . .
.
- 15 -
3~
to receive one-half teaspoon corresponding to 2,5 ml
of this mixture 3 times a day.
The total solution yielded 40 250 ml flasks, each
of ~hich held about lvo dose~ of 2,5 ~1 and therefore
w~s sufficient fo~ a treatment of about 33 day~.
Each teaspoon having a content of 2,5 ml had a content
of o,15 mg sterolins, 2,5 mg PYP and 1,5 ml ethanol.
It should be obser~ed that concentrations of over
o,o75 ~g of sterolins and 1 mg of PVP for each loo ~1
of 60% aqueous ethanol should not be exceeded if
clear sslutions are decired~ i.e., that about o,l875 mg
of sterolins per 2,5 ml of aqueou~ 60fi ethanol is the
maximum dose for a clear solution.
According to the described proce~s the~e c~. also
be produced solutions of sterol monoglycosides or
~onoglu~osides. Of course, all of these compounds
ha~e low ~olubilities so that these solu*ions
according to the described method~ should n~t be
expoced to low temperature~ to pre~ent turbidity.
(b) Solutions containing semisynthetic sit~s~erol-¢-U-
Maltoside.
800 mS of sitocterol-~-D-maltoside uere dissol~ed in a
mi~ture of 3 liters of ethanol and 7 liters of water
at gentle reflllx. The cooled 30% aqueous ethanvl
solu~ion was then filled into 250 ml flasks. The
patients were directed to receiYe 2,5 ml o~ this
solution 3 times ~ail~ (accordin$ to the size thereof
of one-half or an entire teaspoon full).
. , .
':
' . ': ,. ': , : '
.: :
. . . : :
. .
:
-~ - 16
The total solution yielded 40 250 ml flasks, each of
~ic~ held about loo doses of 2, 5 ml, so that ths
total amount wa~3 sufficien* for a trea*m~nt of about
33 days. 2,5 ml of the s<,lution contained o,2 mg
sterolins and o,75 ml of ethanol.
According to the de~cribed process there can also be
procluced solutions of other ~terol dis~ccharides.
The water solubility of the ~-D-maltoside, the
~-D-lactoside, and the ~-D-celIobioside of sitosterol
are respectively o,38 mg, o~21 mg and o, 75 mg psr ml
of ~ater at a temperatuxe of 24 C. This soIubility is
a~ove the preferred individual do~es employed of the
compounds.
p~ e~, recA,
The ~IY~ Y~i~ i~dividual dose employed for st~rol
disaccharides is o,2 mg, therefore o,'~ m~ per day.
It-is therefore indicated that o*her pharmaceutically
e~fecti~e co~pounds can ~lso be worked into the solution~
together with the ~terol disaccharides. Furthermore, the
; alcohol content of these sterolin solutions can be
changed; in a given case alqo other pharmaceutically
unobjectionable solvent~ can be used. Furthermore, ~
there can also be used pure water as the sole solvent.
.
According to the described method the spiroketal-
steroidglycosides can also be wor~ed up into pharma
cautical'y acceptabl~ solutions.
_xample_5
; Pharmacological testin~ o~ sterolins and spiroketal-
stero d5lycosides.
(a) Toxicit,~ testin~_of s~erolins.
In the testi11$ of the ac~te toxicity with rats, mice,
,
, . . . : , . : :
, : .. . . : ~ :
:
:
rabbits, dogs and primates by oral administration of e.g.,
sitosterol-~-3-glucoside even in doses of 1 to 2 grams/kg of body
weight no toxic effects were ascertained. Also, in administration
over a long period of time of daily doses of 100 to 200 mg/kg of
body weight with these animal species there could not be detected
toxicity and also no gout-like appearances so that the tolerance
can be designated as good.
Example 6
Proof of activity of PGS inhibitor
The activity of the compounds as PGS inhibitor was proved
according to the method described by A.L. Willis. Conditions of
trial are described for example in Proceedings of a Workshop held
during the VIIIth European Dermatology Congress Helsinki 1975.
Siliconized cuvettes of the aggregometer are used at a
temperature of 37C as incubation vessels in which an arachidonate
solution was rapidly stirred with a PG synthetase enzyme system,
usually from sheep vesicular gland. To this solution in the
cuvette there is added anticoagulated platelet-rich plasma,
previously warmed to 37C. Light trarlsmission through the cuvet-te
was recorded immediately after addition. The comparison sample
showed a significant peak in the platelet aggregation after 45
seconds of incubation time which proved the formation of PGE2
and PGF2 and the corresponding platelet aggregation.
In the test samples containing 0,00001~ sterolglycoside
-17-
,
, .
. ~
~ ~ .
;.' . , , ~:
'
:
- 18 -
~ $
or spiroketal-steroidglycoside no platelet aggre$ation
occurred. This clearly shows that the formation of
PGs via endo-peroxide compotmds from arachidonate
was inhibited.
Example 7
It had previously been established that PGs, PGE2 and
PG~2~ as t~ell as the endoperoxide precursors play an
important role in the initiating of rheumatoid arthritis.
Therefore, the activity of the compoullds of the
invention as PGS inhibitors had been tested in
pharmacological trials.
When comparing arthritis based on experim~n~al erysipeloid
of for example r~ts with human rheun~ati~ arthr:itis an
extensi~e correspondence of the morphological c~anges
can be observedO The tests had been ~ade according
to the publication of Schulz et al. BeitrOPath. 154,
1-26, 27-51 (1975). The a~t'hritis of rats caused by
erysipeloid can be reproducl~d with nearly loo ~ercent
; effectiveness ~ith a single injection. In case
of arthritis caused by erysipeloid always more than
6 joints of the limbs are ~ransforlned and namely big
and small joints of the limbs in the sam manner.
~ith experimental erysipeloid all animals exhibit
extensively proliferati~e transfo~mations eYen after
3 months.
. ,
In the test of the compounds the following parameters
of the mani~`estation~hase had been ~sed for
estimating the actlvity oi the compound:
.~ ;d
'
.
'' '''
' ~ '
.' ' '' " ,' ' ' ' .
. ~ , .
''.
:
- 19 -
$
aw volume
The arthriti~ of rats i~ characterized in animals
with a body weight of 150 g after the third day,
with a body weight of 200 g after the 5 h day by
an extensi~e periarticular edema.
Xidney (protein secrection)
Nephro~es caused by r.licrothrombes can occur in
30 to 40 percent of the animals during 7 to
8 day.
(
Inflammations of the cornea ~turbidity~ about
8th day
;
xternal genital~, tip o~ tail
Necro~e-~ cau~ed by thrombes about 6th to 8th day.
Aorta
With rats fibrine-rich thrombes occur about 6 h to
11 day at the intima of the aorta. The largest
spac~ covered i~ about 8~h day.
: .,
... . .
: : .. .. ., .. : . .
:
,. . :,:
Male wistar rats with a body weight of 150 to 180 g were
used for the tests. The animals had been kept in single cages and
received a rat standard diet (Ssniff R) and water ad libitum.
The room temperature was constantly 22C, the relative
air humidity 50-60%. The daily illumination time was 12 hours.
Prior to the beginning of the tests the animals had an
acclimatization period of 10 days.
The infection was carried out with the erysipeloid strain
T 28. The dosage was 2 ml s.c. (about 100 200 millions germs).
The compounds to be tested had been suspended in sterile saline
solution and had been injected i.p. in a dosage of 5 mg/kg body
; weight. The treatment was 5 times per week from infection day
to end of test or death of animal.
The evaluation of the characterizing aorta thrombes
showed the following results
sitosterolglucoside: 2,00
hecogeninglucoside : 1,38
diosgeninglucoside : 1,33.
.
-20-
. , ' : ` '
