Note: Descriptions are shown in the official language in which they were submitted.
CASE Y ~I P
~his invention relates to the prepara-tio:n o-~ verbe-none~
myrtenal and pinocarveol starting from oxidized terpene mixtures~
and their use in the therapeutics of bronchopneumonial d.i.seases.
The preparation of verbenone and myrtenal starting from
alpha-pinene by oxidizing self-catalysis in the presence of salts
of chromiumg cobalt, and o-thers has been repeatedly described with
yield of from 15% to 20%o One of the reasons for -the low reaction
yields is the formation of side-products of the homolytic reaction
(radicalic reaction) such as verbenols and myrtenolsg these latter
being alcohols which cannot be further oxidized to the ketones in
the specific reaction medi~mO
In the Swiss Pa-tent Specification N 542 163 of the same
applican-ts hereof~ a method is described for the preparation of ter-
pene frac~ions which are useful in -the treatment of bronchopneumo-
nial diseases~ said method being based on the oxidation o mixtures
which predominantly contain alpha-pinene~ More particularly, with
the method according to the above mentioned Swiss patent9 two
terpene fractions are ob-tainedg~which~ dis-till.grespectivelyg in the
temperature ranges of from 40C to 600C and of from 650C to 103C.
It has now been found9 and this is the subject-matter of the
present inventiong that such ox1dation mixtures can be subjected
to an additional treatment whichg in the case of the preparation
of verbenone and myrtenalg is a fur-ther oxidation -treatment to be
carried out with chromium trioxide in sulfl1ric acidg whereas~ in
the case of pinocarveOl , it consists in opening the oxyrane ring
of the alpha-pinene epoxide and subseqllent -transposition of the
resultant compound~
It has been found, in Çac-tg that in the case of verbenone
in
and myrtenal~ the oxidizing treatment with chromium trioxide in
sulfuric acid of the oxidation mlx-tures of the Swiss patent afore-
mentioned, the oxidative method goes on with a totc~l disappearance of
'~
9~
the alcohols and a formation of ea~bonyl. compounds (ketolles and
aldehydes)0 Upon treatment with chromiLLm -trlo~i.de in sulfuric
acld an :isolati.on of -the as-obtained ketone bo~'L,es is accomplish~
ed 'by a bis~Llfite complex and -ractional distillation.
In the case of pinocarveolthe opening of the oxyrane ring
is preferab].y carried out with alu.mi.n~ isopropoxide~ whereafter
the removal of the carbonyl compounds and the fractional. distilla-
tion are proceedecl with in o:rcler -to isolate pinocarveolO
As regards the therapeuti.cal u.se of the compounds obtained
with the me-thod according to the present inven-tion7 the thera-
peutical use of the terpene fractions di.sclosed .in the Swiss patent
542 163 is already known9 ancl more particularly the use of the
fra,tion which clisti~s in -the 65C-103C range (t~ be indicated
hereinafter as the "frac-tion 2"~o
The balsamic ancl analept.ic actlons of such fraction 2 are
already known9 the therapeutical use of such rac-tion be.ing just
due to such actionsO
- In addi.tion -to the uncertainty from a scienti.fic standpoint
as to the exact identification o th.e acti-ve principles which are
responsible for the therepeutical aetivity of pharmaceu-tical compo-
sitions based on such fraction 29 the drawback is apparent9 from
an industrial viewpoint~ of working on a mixture of compounds having
a eomposition whieh is ehangeable9 even within a certain rangeO In
addition~ the fact is worth noting that the pharmaceutical composi-
tion containing the fraction 2 has a general. indication as a 'balsa-
mic~ along with a few side acti~i.ti.es9 without any possibility being
afforded of-exal.ting bhe one of these acti.vities p~eferably over
the others3 or of imp-roving the general activi.ty~
It has now heen found~ and -th:iæ :is anot.ller as~)ects of the
present .inventlon9 tha-t -the te:rpene compoLmds in question po~ssessg
in additi.on to a~ome alrea~y known actionæ~ su.ch as th.e 'balsamic
and the analeptlc ac-tion.s~ also apprec~.i.a'b:l.e bronchodilati.ng~ antl-
z~ ~
phlOgisticg artiexudat.ive and a:ntiaggregative~ ac-tionsO
The demonstration of -the pharmacological ac-tivity of ver-
benoneg pi~ocarveol a~d myr-ter~al is based o-~. the fol,lowing para -
meters :
1. Acute toxicity
Act.ion o~ t:h.e bronehial. muscles
Ac-tion on the in-flamrnatory process
4 0 Action on haemolysis
Ac-tion on th.e thrombocyte aggregation
Ant.ibacterial action~
ln all these -teOEts5 -the acti~ity of' the diferent compounds
has been compared,wi-th that of the -raction 2 as def'ined above
and with that of therc~peutical preparatïons based on terpene alcohols~
more particularly sobrer~la
The individual compounds the subject of the present invention
will ~e now consi.dered :
A - Verbenone
1~ Acute Toxicit~ O ~
TABLE 1 reports the LD$o (and the -fiducial limits 95~ as
determined accord.ing to the Lltchfield and W:ilcoxon method (JO Pharma-
col. Expo TherO~ 96~ 999 1949)g by admin~ster~7ng verbenone both
intraperitoneally (i~po ) and orally (p~O) to groups of 10 mice and
6 rabbits per dose~
T A B L E
COMPOUND Animals Sex Administra- LD Gleason classi-
tion route mi~igrams fication
~" W o
Mice mal.e iopo 361 (337~-385~ Poor
fema:L _ ~,,_~ ~"_ ~,_ __~
VERBENONE Mice male pOo~ 1410 (l.'L6$~1706) 51:i.gh.t
3 female 1,530 (1124-1912)
~ O
Rabbit~ ma~Le i~po 3l~ (250-384) Poor
female 232 (l97-274)
Rabbi-ts rnale pOOD 790 ~608-1027) Poor
female 65.5 (57~-/533
On the basls O:e -these data -the verbenone appears a well
tolera-ted eompound~ since its toxlcity is of the slight or poor
magnitude aecording to the Gleason classi.-fieation~ as related to the
anim.al spee.i.es and the administration route~
2~ Bronchoclilat,i~LfL~
The verbenone has exhibitedg both in vitro and l vivo a
pronounced bronchodilating ac-tîvity9 by vir-tue of which it distin-
guishes in a statistically significallt way both o~er the other medi-
einaL subs-tances present in the fraction 2 o:E the SWi3S patent
542 163 ancL over the referenoe terpene3 .ioso sobrerol~
a - in vit_
On the Guinea pig traehea~ isola-ted aceordlng to the techni.que
by Costantine (J~Pharm.O PharmaeolO9 l7~ 384~ 1965)g the verbenone
hrings about the relaxation oE the smooth tracheal muscles a-t the
eoneentration of 125 to ~000 mierograms per milli.li-ter with an
20 intensity whieh is sig~ificantly higher than that of the terpenes
of the fraetion 2 and also than th.at of sobrerol (TABLE 2)~ In
add.itiong at coneentrations in t.he range of from lV 3M to 5010
it inhibits the histamine i.nduced eontractions o~ the isolated
Guinea-pig trachea (Histamine 10 M~ see TABLE 3 3 o
T A B L E
PereeIItages of re.Laxat.ion (a~erage -; standard de~iat.ion of 4
preparations3 of the .isolated Guinea-p.i.g traehea as brought about
by eoncentrat:ions of L25-.500 and 2~0~0 mierogrrarrls per m:illiliterO
~Og~2~
COMPOUND :P~eparation3125 lug/ m3500 jLg /ml 27000 ~g/~ml
VERBENONE aveO 26025 61~25 71037
stdOdev~2~39 2039 1015
F`RACTlON 2 ave~ 1305 33~75 52025
4 stcl"devO 1070 11,4C) 2~.95
SOBREROL aveO1087 5062 20~75
stdDdevO 1019 2013 1,49
_A B L E
Verbenone percentages of inhib.ition of the histaminic spasm
due to rela~a-tion o-E the smooth tracheal muscles ~average ~
standard deviation of 4 preparations~ :
CONCENTRATION 10-3~ 501~ 3M
.Av.erage 2902 9608
Standard devia-tion 501 1696
. _
bo in vivo
The verbenone injected intravenously in anes~thesi~ed dogs at
dosages of -from oO6 milligrams per kilogram body wei.ght to 408
mgs /kg/how~ produces a pronounced recluction of the lung resistances
as determinecl accorc~ng to the techniql~e by Diamond ~ArchO Int~
PharmacodynO~ 168g 239~ 19673 ~see TABLE 4). In addition7 verbe-
none perfused intravenously in rabbits at the dose unit of 2~5
mls per kilogram bowo per minllte~ inhibit,s lih~ histamine
induced experlmental bronc,hospam (~:Lstamine lOO mLcrog:rams per k.ilo~
gram bJw~ intravenously) to a de~ree ~:hi.ch is sta-t.is-ticall.y hi~he.r
~g2~
than that of the terpenes o~ t~le fract.ion 2 ancl also -t:han -tha-t
o~ sobrerol (TABLE 5)~0
T A ~ L E ~
VERBENONE ~- Percentage red~ct:ions of the l.urlg Resist,ance by
intravenous admini,stratlon to dogs (Average l Standard
devi,ati.on o:r 4 an.imals ,)
Do3age
mgs /kg bow~ o~6 1~ 2~4 408
Ave.rage7~175 lOr85 220225 3006
Standard
deviatlon60786 7~;7 180636 130507
l o T ~ B L E _5
Percentages of inhibîti.on of the h:istamine-lnduced broncho-
spasm by intravenous perf-usion i.n rabbi-ts (ave,rage of 4 animals '
stand.ard deviation)O
Per~usion times
COMPOUND 15 30 . 6 0
minutes m:inutes minutes
~____ _ _ _
VERBENONE average 36 ~ 2.54& 0 27 7 3 ~ 3
stdOdev~, lo65 9000 7017
. ~
FRACTION 2 average 16~,30 24.~20 38~92
s-tdOdevo 3~62 4039 3~67
., , . _ , _ . _ ~ n . . . _ ., v . . . ~
SOBRERO:L a-verage 2ql5 L ol5 3~97
~ stdqdevO 403C) 603~ 8tl73
3o An~i~inflammator;y ~ctivity ~ -
In albine rats of the COBS (Charles R.i.ver) stoek~ verbenone
as administered intraperito:nea,l.1y at the dosage o~' 30 mi.ll.igrams
per k;logram bow J ~ in'hibits t'he experimen-tal oedem~ in th.e paw as
~ ~,
~39~
induced by carrageenin (Winter~ C~AO et al~ ProcO SocO Exp~
Biol. MedOg 111~ 544g 1962~)both in normal rats and in su~rarenal-
ectomized rats to a degree which iS9 Erom the sta-tistical stand-
point, significantly improved over those O-e the rac-tion 2 and of
sobrerol (TABLE 6)o
When adminis-tered intraperi-toneally in rats at the dosages o
36 and 120 milligrams per kilogram b.w~ verbenone displays an
intense antiexudative activity in the experime:ntalJ t~rpentine -~-~
i~duced pleuritis (Hurley~ JO VO et alOg J~ PathOg 91~ 57531966) to
a degree which can be compared to that of aspirin (TABLE 7)0
T A B ~ E_ _6
Anti~inflamma-tory activity on the carrageenin-induced
oedema in ratsO ~
CO~POUND Dosage milligrams/kg bow~ % inhibition of plantar
intraperitoneally oedema~ as after 4 hrs~
from adm.inistration
(ave~+ S-tdOdevOon 6
ani.mals
..... .~ ... __._ _ _ . . . __. ~ ___
VERBENONE 30 29
FR~CTION 2 3 21
SOBREROL 30 11
., .... ... _ . . ~ .. ...... .
Haemol~ytic activit~ in vitro O -
Verbenone protects in vitro the red blood cells o e rats -rom
the haemolysis as induced by capillary-active-agents (Tween 80)
with an effective concentration 50% (EC 50) of 63905 micrograms
per kg bowo (fiducial limits 95~ : 51800-760011)~
Antia~r~ative activity~ in vitro ~ - ~
Verbenoneg at conc~3ntrations of from 160 to 1.280 m.icrograms/
milli3.iter inhibits the thrombocyte aggregation Erom ADP in vi-trog
evaluated according to the me-thod by Born and Cross (J~ Physi.ol~ 7
Londong 168~ 178~ 1963) to a degree which is higher t:han that Oe
the terpenes of -the Fraction 2 and of sobrerol (TABLE 8)o
T A B L E _ 7
Activity of Verbe:none on the Turpentine-Induced
Pleuri-tis in Rats O -
Intraperitoneal Exudate~ + StdODevO Inhlbition Signifi-
administration mls 3 ~0 cativity
Controls 1097 0024
(solvent)
Aspirin 1030 0013 34 % ~ 0~01
(100 mgs/kg)
Verbenone 1023 0024 3706% ~ OoOl
(36 mgs /kg)
Verbenone 0097 0022 50~1% .~OoOl
(120 mgs ~'kg~
T A B L E __8
Thrombocyte aggregation inhibition percen-tage (average
of repeated -tests)O-
FINAL CONCENTRATIONS % INHIBITION
.
CONTROL o
-: VERB~NONE 1280 ~1g/ml 100
'` " 640 " 44
" 320 " 6
" 160 ll 0
____~
CONTROL
SOBREROL4000 " 32
" 200~ " 2~
" 1500 " 12
Il 1000 ~' 4
CONTROL O
FRACTION 2 1280 " 70
" 1016 " :l8
" 806 " 14
" 640 " O
9~
z~
6 D Antibacterial activit~
Yerbenone possesses a poor antibacterial activity on the
Gram-positive and Gram-nega-tî.ve germs~ w.i-th a MIC (Mirnimum
Irlhibitlng concen-tratioll ) of 800 micrograms/ml on
_~eus and Escherichla coli (TABLE 9) and is as active as
Fraction 2 ancl more active than sobrerolO
T A B L E 9
Minimum Inhibiting Concentrations (MIC) in micrograms per
milliter . -
Mieroorganism Staphylococcus Escherichia coli
Compound aureus
VERBENONE 800 800
F`R~CTION 2 ~00 800
SOBREROL ~
Summing up~ the compo-und has proven -to possess a pro:nounced
bronchodilating action~ by virtue of whic:h it distinguishes in a s-ta-
tistically significant manner both over the other m.edicaments pre~
sen-t in ~raction 2 of Swiss Patent 542 163 and -the re-ference ter-
pene (sobrerol)O
On aecount of these partieular properties~ ~e~benone is efect
ive as the active ingredien-t oE a pharmaceutical preparation which
is especially adapted for the trleatmen-t of bronchopneumonial
diseases which are accompanied by an obstructiorl of -the respira-tory
ehannels due -to in-flammation and i.nfectious causati.ve agentsO The
dosage envisaged for therapeu.t:i-cal appliea-tions is irom 10 -to 100
milligrams dailyO
B - MYRTENAL
lo Acute to icit~ O ~
The I.D5n as determined accordi:ng -to the methocl by I.i-tchfield
and Wilcoxo~J PharmacolD Expo TherO~ 969 99~ 1949) by aclm:inistering
10"
23.~
the myr-tenal întravenously -to groups of 10 mice was 170 milli-
grams per kilogram bow~
2~ Bronchodilato~ etivit~
___
Myrtenal has shswng both l vitro and i.n vi-vo a fair broncho-
di.l.ating activity~
a - in v _ro
On the isolated Guinea pig trachea according to t:he tech-
ni~lue by Costantine (J~ ParmO Pharmacol~ 17~ 384, 1965 )3 myrtenal
determines the relaxation of the -tracheal smooth muscles a-t t:he
concen-tration of 125~200 micrograms/ml. with an in-tensi-ty which is
s-tatistically higher than -that of -the F:raction 2 and that o-f
sobrerol (TABLE lO)o
T A B L E lO
Percentage of relaxation (average ~ standard deviat:ion of
4 prepara-tions) o-f -the isolated Guinea~pig trachea as determined
by concentrations of 125~500 and 2~000 micrograms/milliliterO
Compound Preparations 125 500 2~000
Nmicrogr/ml microgr/mlm.i.crogr/ml
MYRTENAL 4 aveO 7~.S 4'?o5 ~2~5
stdOdeva 500 1500 2007
aveO13~5 33075 52025
FRACTION 2 4
stdOdev~1~70 1.49 2095
aveO 1087 So62 201~75
SOBR~ROL 4
stdOdeY~ 1019 2~13 1~49
b _- iD VlVO
~ yrtenal perfused intravenou.sly in rabbits at the dosage o-f.
205 millil~t~rs per kg bow~ per m:inute inhibi.ts -the exper-Lmental
hi.stamirle~indu.ced bronchospasm (hi.stam.ine 100 m:ic:rograms/kg b.w.
3 intra-veno-usly~ in a manner wh.i.ch is statis-tically ~ ,rher than tha-l;
of sobrerol and slightly lower -than -that of Fraction 2 (TABLE 11)o
T A B L ~ 11
Percentages of inhibition of the ~i.stamine induced broncho-
spasm by intravenous perfusion in rabbi-ts (average on 4 ani.mals
+ standard deviation)O
Perfusion times 15 mins, 30 mins~ 69 minsO
Compound
,, . ~
MYRTENAL ave~ 25~4 2004 28,8
stdf~devo lOr4 1602 13~5
10 FRACTION 2 aveO 16030 24~0 38~92
std,devO 3062 403~ 3067
SOBREROI, ave~ 2~15 1~l5 3~97
s-td~devO ~0~O ~o39 8~73
3 Anti-inflammat~ ac-tivity O -
In albino rats of the CO~S ~Char1.es River) s-tock9 myrtenal
as administered intraperitoneally at the closage o-f 30 milligrams
per kg bowo inhibits the experimental paw oedema from injec-tion
of carrageenin (Win-ter9 A~CO et alOg Proc~SocOExpO BiolOMedO 111~
544g 1962) to a degree which is more intense than that of Frac-tion
2 and also than that of sobrerol (TABLE 12) 0
T A B L E_ 12
Anti-inflammatory activity on the carrageenin oedema in rats~
Compound Dosage % inh:ibiti.or. on the plantar
mg /kg bowo oe.dema aft~r 4 hou:rs as from
intraperito~ administration (average on 6
neal~.y animals + stdo deviat,ion)
MYRTENAL 30 30
FRACTION 2 30 ~1
SOBRE,ROL 30 11
120
92~
Antihaemolytic acti~ n -vitro
r
Myrtenal at concentrations o~ from 80 to 5~0 micrograms/ml.
protects the red. blood cells of rats from the :h~e~olys.~s as i.n-
duced by capillary active agents (Twee:n 80) wi.th a.n Effecti~e Co.n-
centration 50% eq~al -to 157009 microgr~ms/ml (fiducial limits
95% -- 11 2o3 ~ 18~o9)~
Antiae~re~atlve activi-ty . ~
Myrtcnal at concentration~ from 160 t,o 1~280 m.icrograms/ml
inhibi.ts i vitro the thrombvcyte aggregation -from ADPg e~alua-ted
according to the method by Born and Cross (J. Physiol~g Lond.ong
168~ 1789 1963)9 to ~ degree which i3 h:igher than those of F'raction
2 and sobrerol~
T A B L ~
Thrombocyte aggregation o % inhibi~,lon (average o-f repeated
tests)
FINAL CONCENTRl~TIONS % INHIBITION
CONTROL
MYRTENAL 1280 jug/ml 75
~' 6~o ll 20
" 320 1~ 12
11 160
CONTROL O
SOBREROL 4000 ll 32
2000 ~ 2
~' 15 oo 11 1 2
Il looo 4
.. -CONTROL o
FRACTION 2 1280 " 7
1016 ll 1
130
2~9L
FR~CTION 2 806 ' ~g /ml 14
" 640 " O
~_
Myrtenal possesses a pronounced antibac-terial activity on
Gram-positive germs~ less,intense on Gram-negative ones with a
Mi.nimum Inhibiting Concentration (MIC) of 200 micrograms/ml on
Sta~hylococcus aureus~ and of 800 micrograms/ml. on Escherichi.a
coli and is more active both than Fraction 2 and ,sobrerol.
(TABLE 14)o
T A B L E~
M:inimum Inh:ibiting Concen-trations (MIC) in micrograms/milli-
liter O
Microorganism ~ ylococcus Escherichia
Compound aureus coli
.. _ . ",_ _ .
Myrtenal 200 800
Fraction 2 800 800
Sobrerol ~ v~
. -- ~
This compoundg myrtenal~ has shown~ by way of conclufiiong that
it possesses a marked antibacterial action by virtue of which it
20 distinguishes in a statistically significant way from both the
terper~mixtures of Fraction 2 of Swiss patent and the other medi-
cinal substances o-f referenceg Myrtenal has an anti in.Elammatory
action of poor magnitudea it displays an antihaemolytic activity
andg fi.nally~ it has also a bronchodilating type acti,on and an anti-
aggrega-tive action as well~ ,
On account of' these particu1ar features there~ofj myrtenal is
indica-ted as an active ingre~.i.ent of' medicinal compositions which
are particular'ly sui-table for the therapy of 'bronchopne~lmonial
diseases when an importarlt bacte:rlal. component is presen-t~ or wheng
1~
at any rate., an anti.bio-tic~-based therapy is requiredO In con-
nection with such i.ndications~ -the a~linistrat.i.on of a daily
dosage of' from 10 -to 100 milligrams of myrtenal ls suggestedO
C - PINOCARVEOL
Acute toxici~
~_ ~
The ~D50 as determined according to the method by Litchfield
and Wilcoxon (J'O PharmO Exp~ Ther. 96, 98g 1949) by adminîstering
p.inocarveQl intravenously to groups of 10 rats per dosa~e was
equa.l -to 140 millig:rams per k:ilogramO
20 Bronshodilating act vity
Pinocarveol. has exhibited~ both in_vi-tro and in vivo~ a fair
bronchodilating actlon~
a ~ in vitro
On the G~inea-pig trachea isolated according to the technique
by Costantlne (J~Pharm. Pharmacol.0~17~ 384~ 1965)~ pinocarveol
determines the relaxation of the smooth tracheal muscles at the
concentrat.ion of 125-~oOOO micrograms/ml wi.th an intensity which is
statistlcally hîghe.r than that of sobrerol and which can be compared
to that of Fraction 20
b - in vivo
Pinocarveol perfwsed in-travenously i.n ra'bbits at the dosage
of 205 milligrams per kilogram and per m:inute ~ the
experimen-tal bronchospasm induced by histamine (100 micrograms
per kilogramg intravenously) in a manner which is statistical-
l.y hi~her than those of FRACTION 2 and of sobrerolO
3. Anti- ~
In albi.no rats of the COBS (Charles R.i~er) stockg pi.nocarveol
adminis-tered intravenously at the dosage of 30 milligrams per kg
inhibits the experLmental paw oedema from injection of carra
3o geenin (Winter9 CoA~ et a~ Proc. Soc~:ExpOB,Lol~ Med~ l'L'L, 544~
1962) to a degree whîch is more intense than those of Fraction 2
and of sobrero.LO
Z~
ctivi..t~ ~vitro~-
Pinocarveol at concentrations of from 20 -to 200 micro-
grams/ml protects the red blood cells of rats from the haemolysis
cau.sed by cap.il.Lary-ac-tive agen-ts (Tween 80) wi-th an Effective Con~
centra-tion 50% (EC 50) of 132~29 micrograms/rnl (F'iducial limits
95% - 111~5~153~0~)o
s,~g~.ive ac-t:Lvity ~ -
Pinocarveol at concentrations of' from 160 to 10280 micro~grams/ml inhibit.s9 l _ .itroJ the thrombocyte aggrega-tion due to
10. ADP~ evaluated according to the method by Born and Cross (J~ Physiol..
London~ 168a 1783 1963) to a degree which exceeds t:hat of the ter-
penes of the Frac-tion 2 and that of sobrerol~
Anti-bacterial activlty O -
Pinocarveol i..s endowed with a poor an-tibacterial ac:tivity
both on Gram~posit~ve and Gram-negative germs with a MIC (Minimum
Inhibiting Concentration) of 800 micrograms/ml on Staphyloooccus
aureus and Escher-ichia coli and is thus more active than sobrerol
and as active as Fraction 20
Summing up, th.e compound pinocarveol has proven to possess a
pronounced anti inflammatory aCtiOn by virtue of whi.ch it distin-
guishes in a statistically signifLcant manner from the other medici-
nal subs-tances such as the mixture of Fraction 2 of the Swiss Patent
aforementioned and the reference terpene(sobrerol~)~
P:inocarveol possesses a poor bronchodilating action and is also
endowed with a pronounced antihaemolytic activity9 by virtue of
wh.ich it is possible -to attribute to such compound a stabilizing
action at -the cel.lular membrane level~
Pinocarveol displays a poor antibacterial action and also
exhibits a consp:icuous anti.aggregative ac-ti,onO
On account of its particular properkiesa pinocarveol can. be
the active ingredien-t of a medicament wh:i.ch i.s particularly su:it~
able for the treatment of bronchopneumonial diseases wh.ich are ac-
16~
%~
companied by a strong :inflamma-tory pa-ttern of -the respiratory
channelsO
The dosages to be recvmmended for this therapeutic appli-
cation are from 10 to 100 mi.ll.igrams dailyO
The pha~maceutical compositions according to the present
inventi.on can be presented in the form of preparations f`or oral
a~ninistrati.on9 normal or delayed~actiong such as soft capsules~
or injectable ampoules, suppoæitories~ sprays in various solution
forms~ ointments and balms with the usu.al media~ fillers and so on
as conventionally used in the pharmaceutical. art~ both as individual
components and iIl association with medicaments indicated in the
diseases referred to above9 that is~ antibiotics, antibacterial
subs-tancesa chen~therapeu-tic substancesg sulfonamide drugsg anti-
inflan~Ia-tory d:rugs, cortisone preparations and analgesicsO
The following examples are intended to illustrate without limi-
tation the preparation of the compounds according to the present
in~entionO
EX~MPLE
____
a - Alcohol oxidation
lUO grams of the terpene mixture of Fraction 2 of the Swiss
Patent 542 163~20~30% oÇ whi.ch comprise~ compound.s havlng a car-
bonyl moiety (predominantly verbenone and myrtenal)whereas 50-60~
comprises compounds having an alcoholic moiety (mainly verbenol and
myrtenol) are dissolved in 2~000 mls of dry acetoneO
Separately~ the oxidizing sol-ution is prepared by admixing
cautiously 37~41 grams of CrO3 wi-th 32~2 mls of concO II2S04 in -the
quantity of water which is sufficient to make up a final volume
exactly equal to 140 mlsO
The chromic acid is then added slowly and dropwise to the ace-
tone solution with stirring~ coo:Llng on an ice bath so as to main-
tain the temperature constantly below 30~Co On completi.on of the
addition~ filtra-tion is carr;i~d out on a Celite (Trade Mark) a:nd
2~
the filtrate is taken up by evaporating off acetone under reduced
pressure at 40C~ The residue i.s then dilu.ted with wat,cr9 neutra-
lized :i.Il cold condl.tions wi-th 10% :NaOH a:nd extracted wi-th CHC-L3
until. exhausting the mo-ther liquors. The combinecl organic extrac-ts
are dried. over anhydrous Na2S04 and evaporated to dryness undcr re~
~uced pressure at 40Co
By so doing~ there are obtained about 100 grams o-f a raw
prod~ct which i3 composed.~ for so-60% by compounds having a carbonyl
moiety (prevailingly verbenone and myrtenal)~
b_ - Se~ ~ m th~ raw materlal as
obtained F:rom the oxidatio~l run 0
An app:ropriate vessel is charged with lOO grams of a raw mixture
as obtained from the stag~, a of this methodg dissolved in 500 mls of
ether and a soluti.on Eormed by 120 grams of NaHS03 and 72 grams of
NaHC03 in 2~000 mls of waterO T:he mixture is vigorously shaken
during abou.t 20 hours at room tempera-ture9 and -the~ -transferred -to
a separatory -fun-.nel and the organic phase is discarded. The aqueous
phase is washed twi.~e with 500 mls of ether each time~ whereafter
the aqueous bisulfite soluti.on is taken up and subjected to stea~
dîsti.llation~ Thc processing of the distil.late gives 50--55 grams
o a mixture whlch is virtually composed by pure verbenone and
myrtenalO
c - Obtention o~ pure verbenone a:nd myrtenal O -
50 grams of the product coming from the previo-us s-tage are
subjected to fractionation in a vacuo at 20 millimeters of mercury~
using a reflu~ column ~illed with nickel shavings9 T:he distillation
is carr.ied out slowly (5 -to 8 m:Ls ~ hou.r~ by maintaining a refl.ux
ratio of about 40 to 1 the whole run throughoutO The -fract:ion
which disti~ at 90C-9.~C is composed by myr-tenal (19 gratns) arld
has the -eollowing physico~chemical speci-~i.ca-t:ions :
n2o -- 1050 d a oO98
D
180
z~g~
Inf:rared Analysi~ : bands at 1684 cm y C=O (conju~ated)
1623 cm. ' y C C (conjura-ted~
14~1 cm y C~H
1423--1387 1372
UltraViolet Analysis ~ (E-tOH) = 246 nm ( ~ about 8~00)
The semicarba~one mel.ts at 210~C-215C~
The fraction whlch d.ist~ls sub~equently at 110C~113~C at 20 ml.lli-
meters of mercury is composed by ~rerbenone (28~5 g:rams~ and has
th.e following physico-chemlcal specl-fications :
~0
10 n = 1~49 d - 0~97
Inrared band~ at 1670 cm 1 ~ C=O (co.njugated)
1615 cm C=C (¢onjugated)
1650-1.435-13~0
IJltraviolet ~ (EtQ~ - 2SO nm ~ = 79300
Semicarbazone : mO poi.nt 1~8C~190C
EX~MPLE 2
rsi.on of al ~ ~ ~
60 grams of the terpene mix-ture of Fraction 2 Oe the Swiss
Patent N S42 163~10-i5~ of which compri.ses alpha-pinene epoxide~
where~s.2,0-30% con~ists of compounds havin~ a car'honyl moiety9 and 50~
60% consi3~ ~e compounds having an alcoholic mo.iety (verbeno1.~pino~
carveol and. myrtenol)9 are dissolved in 60 mls o anhydrous tolueneO
To the solu~ion is added an excess of aluminum isopropoxide (about
2-3 grams) and the mixture is boiled during 10 minutesO The mix-ture
is carefl1lly cooled and slowly aoidifled by cautiously adding dilu-
ted sulfuric acidg while still keeping a-t a tempera-ture of 0C or
belowO The phases are separatedJ the liquors are ex:hausted with
toluene~ the organi.c solutions oosnbined~ wassled w~.th water and
dried o~er anhydrous Na2SO~O Filtration i$ carri.ed out and evapo:ra-
tion at 60~C under reduced pressu:re is e.e*ected untiJ.. a solid re-
sidue is obta.ined~ There are obta.ined about 60 grams of a raw
1~30
;4
product which is exempt f~om epoxide and is enriched with com-
pounds having an alcoholic function (pinocarveol~ verbenol and
myrte:nol ) O
ration of the car'bonyl com~ounds
~,
The residue which has been obtained from the pre~ious stage
is takerl up with 300 mls of diethyl. ether and the resultant solu-
'tiOIl is subjected to vigorous stirring wi-th a mixture ormed by
60 g~ams of NatlS039 36 grams of NaHC03 and 1~000 mls o~ water~
such treatment heing carried out during 20 hours at room -temperature~
The mixture is -transferred into a 3eparatory funnel and the phases
are separated. l'he aqueous phase contains myrtenal and verbenone
.in the form of soluble bisu,l.fi-te adducts~ from which they can be
reco~e~ed in the form of pure products by working accorcli.ng to what
has been .indicated -or the previous stages~ The organic phase~
which contains the alcoho~Lic compoundsg is washed to neu-trality with
water~ dried over Na2S049 filtered and evaporated to dryness under
recluced pressure ~ There are obtained 40-50 grams of a residue
which is predominantly composed by pinocarveol~ verbenol and myr-
tenol.
c - 0btention_of pu ~ O ~
The r~sidue coming rom the prev.ious stage is subjected to a
seri.es of fractional dist.illation under VaCUIlm~ at 20 millimeters of
mercury, using a reflux column filled with nickel shavings~ The
distillatiorl is carried out slowly (5~6 mls / hour) by maintaining
a reflux ratio o-f about 40 1. during the entire runO The fraction
whlch di.s-tî~s at 100C-106C under 20 milli.meters of mercury and
w.hich is strvngly enriched with pinocarveolJ is collected and sub
jected -to redistillation ~mder the same condi-tionsO There are obtain-
ed t5-18 grams approxO of 95%-98~ pure pinocarveol which has the
following physico c'hemica'L ,,pecificati~ns : b~point .103-104C under
nD = 104988 ; d20 = oO98 20 nlm tlg
Infrared analysis : characteristic bancls of terminal double hond at
6Ooo micirons and at 11O20 micronsO
20.,
