Note: Descriptions are shown in the official language in which they were submitted.
13~?3~19
RS CAS_147
~ he invontlon relata~ to GinX~olid~ d~rivAtlv ~, to
met~o~ for their prep~r~tlon, and to phAr~ceu~l~al
~ompo~ltion~ oontalnlng t~em.
The lnvention provldes l-~lkoxy derivatlve~ of
Glnkgollde~ A,~,C,J and M and further provl~-s 10-~lkoxy
aerl~Atives o~ the ~ald GinXgol~des. Th~ pre~errod al~oxy
groups are ~e~hoxy and othoxy groups.
rhe ln~entlon al~o provid~ a ~ethod ~or the
preparation o~ l-alXoxy derlvative~ of Gin~olld-~ A,B,C,J
lo and M and o~ 10-alkoxy derivativoe o~ the ~ald Glnkgollds~,
the method co~prl-ing reactlng, ln ~olution, Glnkgolide
A,B,C, J or M w~tb an exc~ss o~ d$~zoalkAne ~nd -parating
the re~ultant ~lYtur- o~ l-alkoxy-Glnkgol~de and
10-alkoxy-Glnkgolla~.
In a pre~err-a proceduro, the ~oloet-d ~lnXgollde
i6 dl~solved ln dloxan, ~uita~ly at ~ concontratlon Or lg
per 100 ml, an~ the sel-ote~ dlazo~lkan~ 1~ di-~ol~ed in
die~hyl other. ~he olutlon ~ontalnlng th~ al~oalkAn~ iB
slowly a~d-d to that contalnlng the Glnkgolld~, allowing
ten sgu~val~nto o~ ala~oal~an~ per ~qul~ nt of
Glnkqolld~. Iho ~lxed olution i8 toad at a~b~ont
t-mperA~ur~ ~or ~ro~ 3 to B ~ours, yl~lding ~ mlxture o~
l-alkoxy-GlnXgollde and 10~ oxy-Glnkgolldo. The ~opara-
tlon o~ thQ two proaucts ~rom the r~lnlng non-r-~ct-d
Glnkgollde ~ay u~t~bly be achle~ed by ~porAtlng o~ the
~olvent~ and ~lut~ng the rocldue e~rou~ a ~llioa g~l
column u~ing thyl acet~te : hex~ne 1:1 hy volu~e ~
elu-nt.~he resultlng oolutlon 1~ ovapor~t~d o~ and tronted
.
RS CAS 1~7 13~3
- 2 -
~y ohloro~orm which di-~olve~ lO-alkoxy d-r~v~tlv-. The
~aid lO-al~oxy ~oriv~t~ve i~ r-cov-r-d ~om thl0
chlororor~lo olutlon and the r-malnln~ ~olutlon 1O the~
treateid wlth dlethyl ether w~lch glvo~ the l-alkoxy
dorlv~tlve.
~ he ~lkoxy-Glnk~oll~o~ o~ th- lnvontlon ar- o~
~ntere~t ln th~ treat~ont ? PAF-Aooth-r lnduced ~ladie~,
and the lnvent~on accordln~ly al80 provlde- a
phaxmaaeutlaal composlt~on comprl~ln~ a l-~lkoxy dcrlvAtlv~
of Ginkgolldo A,B,C,J or M or lO-~lkoxy dorl~atlv- o~ one
of the ald Glnkgolld-- or ~ ~lxturo o~ two o~ ~ore o~ ~ld
l-Alkoxy ~nd/or ~d lO-alkoxy derlvatlve- ln admlxture
wit~ a phar~acoutlcally ~oceptable dllu-nt or c~rrior.
ThB lnvent~on i~ lllu~trate~ by the ~ollowin~
15 exa~ple~ :
Exam~le 1
l-MethoxY-G~n~soll~e B and lO-M t~oxY-Glnkaolld- B
~ o ~ olution of Glnkgollde a ln dloxan ~lOg~l) WAB
slowly add-d lO oqul~al-nt- of a olutlon of diazo~ thane
ln di-thyl eth-r. I~ ~lxtur wa- tood at a~bi-nt
to~poraturo for ~ hour~, and t~ n -parAt d followlng the
preferr~d -paration procedux4 d -crlb~d ~bove.
l-methoxy-aln~g~lldo ~, t~o otructur o~ whlch W~E
conflrmed by HPLC, wa- obtain d in 66.1 % yiel~ and
2S lo-~Qthoxy-Gin~gollae B wa- obteinoa ln Z4.4 % yl-ld.
r~
RS CAS 1~7 ~ 3~3~
-- 3 --
Example 2
l-E~hoxy-Gink~ollde B ~nd 10-E~hoxY-Cln~qolld- ~
~ollDw~n~ the proc~dure de~crlb~d ln Ex~pl~ 1, but
using dlazo4thane ln pl~ce Or ~l~zon than~
(duration ~ houx~), l--t~oxy-Gln~golld- ~ wa- Dbtaln~d ln
63 2 % yleld and ~o-othoxy-alnkgolid~ ~ wa~ obtaine~ ln
25,7 ~ ylel~
Proceedlng a~ ~bov~, but wlth Glnkgollde~ A ~md c,
th~ ~ollowlng yi-ld~ w-re obt~ln-~:
=~
¦ Ginbgolld ~ ¦ 72 - ~ ¦ 20 1
¦ Ginbqol~de C 59 2 % 30 4 %
TW~ICI~f
The toxlcity of th~ compounds of tho lnventlon has b--n
lS ~sured on ~1¢- by oral rout~
No death W~B not~ce~ ~t ma~lmu~ ad~ niotrat~on do~ to
mlc~
PNM~CO~aY
-
A proof of the ph~r~aooutlcal lnt-r--t of th~
2 0 compounds of tho lnv-ntion ha~ been o-tabll-hod by tho
following pbarma¢-utlcal experl~ont~tlon~
~3tJ~
1) - Inhibition of the platelets aqqreqation on N w Zealand
rabbits.
The experimentation was conducted on platelets with plasma
of New Zealand rabbits.
Blood samples were taken from auricular artery and placed
in a citrate buffer (3.8 % ; pH 7.4) ; blood was further
centrifugated for 15 mn at 1200 RPM.
The tested sample was prepared in DMSO, then poured on
platelets rich plasma for 1 mn, then a dose of 2.5 nM of
PAF was added.
The determination is made on a Cronolog Coultronics
apparatus which determines the transmission percentage
corresponding to the maximum height of the peak before the
desaggregation.
The percentage of variation of the inhibition with respect
to the transmission percentage is calculated (control :
pure DMSO).
This method was described in detail in LABORATORY
INVESTIGATIONS, Vol. 41, No. 3, p. 275, 1979, JEAN-PIERRE
CAZENAVE, Dr. MED., JACQUES BENVENISTE, Dr. MED., AND
J. FRASER MUSTARD, M. D., "Aggregation of Rabbits Platelets
by Platelet-Activating Factor is independent of the Release
Reaction and the Arachidonate Pathway and inhibited by
Membrane-Active Drugs".
The results demonstrate that the compounds inhibit
the aggregation induced by 2.5 nN of PAF. Five tests made
on 5 different rabbits allowed us to calculate the ICso of
the various compounds using the linear regression test.
The values for ICso on platelets have been found as
follows :
13~
Ginkgolide type
and substitution - OCHJ - OC2H5
position
, ~
B 1- 6.6 10-7 1.1 1o~6
B 10- 2.9 10-7 7.2 1o~6
C 1- 4.2 1o~6 8.5 1o~6
C 10- 3 0 10-6 9.~ 10-6
A 1- 4.6 10-6 8.7 1o~6
A 10- 1.3 10-5 6.2 10-4
2) - AnaPhYlactic bronchoconstriction of a Passively
~ sensitized nuinea-piq
Passive heterolog sensitizing
________________ ___________
Male Hartley guinea-pigs (400-500g) were sensitized by an
intravenous injection (IV) of an antiovalbumin immune-serum
rabbit (Cooper Biomédical, U.S.A.). To obtain a
satisfactory anaphylactic response, 24 hours later, the
following conditions of use were fixed : injection into the
penis of a diluted serum (to half concentration
0.05 ml/100 g).
Bronchoconstriction measure
Guinea-pigs were anesthetized with urethan (2 g/kg IP),
then tracheotomized and ventilated by mean of a respiratory
pump (UG0 BASILE) : stroke volume 1 ml/100 g,
60 strokes/mn.
~,
~ , . . .
` ~3~36~3
- 6 -
A pneumothorax wa- don- to abollsh spontaneous r-splratlon
Th- initlal reslstanc- was kept constant at 10 c~ watsr
pressur- accordlng to the method o~ Konzett and Ro~lcr and
th- cxcess o~ alr volume wa~ measur-d with a bronchospa~
tranduc~r ~UG0 BASI~E) connected to a UC0 ~ASI~ r-cord-r
"Gemlnin~ Th- ~ugular veln was cathet-rlzed for lntravenous
ln~ections Th- anaphylactlc shock was lnduced by an
intravenou- ln~ection of 0 7S mg/kg ot heterolog pa~slve of
ovalbu~ine Products wer- given by oral rout-, 1 hour
before the antiqenlc stimulation ln the fora of a qu~my
water suspenslon, at th- dose of 25 mq/kg
~e~ult~
_______
Th- bronchoconstriction induced by ovalbumin was
xpressed ln p-rcentag- of maxlual bronchoconstriction
lS given by clamping of the trachea Tbe results are reported
in th- following table
Glnkgollde type
and sub~t~tutlon - OSH, - OC,H,
po~itlon
_ .
B 1- - 49 7 - 38 3 ~*
B 10- - 54 9 - 36 2
C 1- - 40 1 ~ - 39 8 *~
C 10- - 30 6 * - 23 1 *
A 1- - 32 0 - 15 1 NS
2S A 10- - 25 2 - 12 2 NS
NS Non Significative
Slqnificative
~ *~ Very Slgnificativs
114~ **~ ~ighly Signi~lcative
. .
.
.
~ .
.
~3~3~1~
POSOLOGY
In human therapy, usual doses for per as
administration are 0.5 to 1 g per diem, in tablets or
gelatine capsules for one month.
In I.V. administration, three weekly injections at
0.05 to 0.2 g in isotonic solution, for one month are
recommended.
~,
