United States Patent (19) 3,920,663

Ferrari (45) Nov. 18, 1975

54) METHOD FOR THE EXTRACTION OF 3,224.945 12/1965 Tyler................................ 260/285.5
LYSERGOL, AND ERGOTALKALODS OTHER PUBLICATIONS
FROM PLANTS OF THE POMOEA GENUS
(75) inventor: Giorgio Ferrari, Milan, Italy Merck Index, 8th Edition, p. 228 (1968).
Erge ct al., Chem. Abstr. Vol. 79, Col. 396006
73) Assignce: Simes S.p.A., Milan, Italy (1973).
22 Filcd: Aug. 14, 1972 Voigt et al., Dic Pharmazic, Vol. 26, pp. 494-503 503
(97).
21) Appl. No.: 280,212
Primary Examiner-Donald G. Daus
(30) Foreign Application Priority Data Attorney, Agent, or Firn-Shlesinger, Fitzsimmons &
Shlesinger
Aug. 17. 97 Belgium............................... (.738
Jin. - 17 legium............................... SS 57 ASTRAT
52 U.S. Cl............................ 260/285.5: 260/313. A method is discloscd for recovcring ergolic alkaloids
51) int. Cl.'........................................ C07DS 19/02 from the seeds of the Kaladana plant, the latter being
58) Field of Search....................... 260/285.5, 313. a plant of the lpomoea section of Convolvulaceae,
Calonyction genus (Choisy) Hallier f. species nova.
(56) References Cited The method is a selective extraction with appropriate
UNITED STATES PATENTS solvents, in order to recover both lysergol and chamo
2, 4,306 4,938 Kucssmen........................ 260/285.5
clavine from the Kaladana seeds. Temperatures and
2.SS, 24 97.94 Moore............................. 260/283.5 times of treatmcnt are critical and thc pre?crred sol
2,809,920 Of 9S7 Stoll.........., ......... 260/285.5 vents are chlorinated aliphatic hydrocarbons.
issis iii.56s Rutschmannet al. 260/285.5 13 Claims, 3 Drawing Figures
U.S. Patent Nov. 18, 1975 3,920,663
 3.92 () (63
Sting i? the sects. is licitly tiscil ?tir its cricini :1.
Mt. Et ()) F(). R T E EXRAC)N (). YSER((). it in. whereas the (wler is tiscil its in antipyretic.
AN1) ERGOT Al.K.A.OI)S FR()\1 IAN'S OF TE the 1ccompanying drawings, which show Kala
'()M()A (ENIS dan:1, there are shown:
this inventity in relates to the extraction i? alk:il its s in lic. : twic' i? 11 it; it is tir is tssist will
living it in erg lic strict irc frin it l; it i? the put in c. leaves inl (lowers;
genus. (Convolvuliuccine finily). in G. 2 the sced-carrying capsulae whicn ripcined.
More detailcully, thc prescnt invention rc.latics to lic and
extriction and isolation in thc purc condition of lyser in Fl(S. and 4 the front aid sidic views of a sccd.
gol ind chanoclavine from thc sects of Kaladana, a () respectively.
tlant identific d hy the scientific nanc Calonyction lip it has now heen found that this plant, and more par.
moea (Choisy) Hallicr f..., nava spccics. ticularly its seculs, are particularly rich with crgolic al
Kilatl:tina is thc native namchy which is indicated in kiloids and these can the usefully cxtracticci hy a method
sincintaneous plant (if the subtropical mountainous ter which is comparatively simple and allows, intcralia, to
ritories, more particularly the pre-Himalayan belt (in htain them in a pure condition. .
cli: ). whilst scicintific classification his heen contriver Accytling to the inventi in the Kil: it in seeds tre
sial anti in concrlant heret fire. it is 1 till (viii.; fly it. It grant r is is \ily.
As it in later v? fict in the ultivation ("thiri's :::::it. stricitly exhaustel is the 'll with sist file light
c 1Y is thrus v? tilli:1. 1.N. iliuir in Sivus "rivitt it. it. htt. iii viller retiv ( ;all till (; 11 it is, the 'i'
ited. Calcutta, 1958, page 51, Kaladana is indicated as ci:tivi is repeated until civinplete exhaustini is ill
c irresponding to the scientific tern portica federacca tained, that which is achieved by repeating thc opera
( . ). in threur tyr fur times.
Contrary.wisc, in J. Pharm. Sc., Vol. 56. pagc 77. The fut-stricci rug is then subjected t an extrac
1967, it the context of an article by Cupta and co. it with appropriate silvents for withdrawing the ac
work.crs. it is affirmcd that Kaladana is thc name by 25 tive principles. Especially useful to this end are the
which non can uricata is commonly known. tastly, in halogen substitutcd aliphatic hydrocarbons such as
the "British 'harmaccutical Code 949," page 459, it chloroforni. carbcn to richloridc. methylcnic chloride
is affirmcd convcrsely, that Kaladana is conposcd by and so forth. in order to facilitatic the cxtraction, the
dricci sceds of porcea liederaceu. halogen substitutcd aliphatic hydrocarbon can bc sup
30 plenientcd by a small percentage of a low molccular
In ordcr to dissipatic any possible doubt, it is dccmcd weight aliphatic
?itting to describe it more comprehcnsively. alcohol and a snal announ (if an hy
Kaladana is of intercst in thc prescnt invention is : droxide. Atch is such is nethnol, ethi ncil. iscipropa
plant which certainly belongs to the Convolvulaceae suitect in act hyrexitics such is an incini: ;re very well
family, section ponocac and genus Calonyction, S can attai to this purpose. The annu its (if alcohol alticul
which cxhibits thc following ?catures: 15% whereas thc alkali netal bases can
reach
A lianous pcrennial plant having voluble twines drocarbqn. The 5% by volunc with respect to thc chlorinated hy
which arc very long, branch-like, herbaccous but extractions of the matcrial arc carried
wood-like hardcncd at the base, furrow.cdhy two longi lit. preferably. at a temperature compriscd between
O'C and 50'C. Thc nun her cf cxtractions which arc
tudinal grooves which arc abundantly fitted with short 4t required for a complctic cxhaustion of thc alkaloids of
hcrbaccous thorns, thcsc bcing slightly curved down the drug is from three to five.
wards and having an obtusc apex. non-stinging. The extracts as obtained arc subsequently cvapo
Akin to C.ntarica ftern (L) G. Don, it difficrs thcrefrom r:tcd to a volumc which is approximatcly cqual to one
for thc caves which are alternatic and chordatc-incumi tenth of thc (riginal one at a temperature of 30'C and
natc. incit her lobatic nor sagittatc. having slighly sinua 45 under stihatmospherical pressurcs and allowed to stand
cundulatc edges and a greatcr sizc:lcngth up to about a ?cw days at O'C-4C. Thc nass which separatcd after
25 cms. and width up to 22 cms, the petiolc being long this stay is removcd by filtration. The precipitatic cakc,
from 4 to $ thc length of thc ca?, shortly expandcd at in turn, is sturried in ten times its weight of watcr and
thc basic and scarcely hairy there. thc portion which is insoluble is dissolved in methanol
Thc flowers are in number of 2-3 on an axillar pe and combined with the filtratic. The filtratic is washed
duncle; the coro is clongate conical tubular, whitish, 50 with watcr.
having a length shortcr than that of C.m. uricattent From the organic phasc. which is washcd with water.
(about 7 cms.), funnel-likc rotated edge, which is obu thc cxtraction of ergolic alkaloids is procccded with hy
uscly ential hate antt has a diameter i? about 8 cms. using at quick is solutions of acids. it. has hecn ?ound that
lic colour heing lilitic or risy, intre (nr less if k which the tiquettis diluted stilitions (if phosphoric acict litici
it is a night time; st: it in a tail stylus lt in it if it fitte ticiselves very well title task. The extrict it in f the
frt it the civil title, the calix his incun inite it litti. it: itic phase with attick is solutions in?: cils is contin
cite it thes, which towever hect: ct; recal intil hi:- tied until the Ethrlich alkaloid test lectivities negative.
conting ovatc-acuminatc and slightly divergent as the From the combincd acidic extracts, which are made
capsule ripcins. “t weakly alkaline with a hase (preferably aqueous ammo.
The capsule is pendulous, ovatc-apiculate laving ini: ), the alkaloids are extracted again with a mixture of
throc ( from two to four) black scc.ds in thc form of a an aliphatic chlorina cd hydrocarbon and an aliphatic
clovic (outer fice voidal and the int crnn fic cs pla alcohol. A nixt urc of chloroform and mc thanol in the
nar), minutcly tomcn to us-granulous, long 7 mms, as an ratio of 7:3 by volumc proved to be particularly satis
averagc and wide 6 mm s. 65 ?actory.
As to its habitat, thc plant is spontancous in the Thc extractions arc rcpcatcd until the Ehrlich tcst is'
misuntainous pre-Himalayan helt of India and Pakistan negative, The combinca cxtracts arc washcd once with
(East), up to an altitude of 500 meters. Thc drug, con water, tric diver in hydrous sodium sulphatic and then
 3 3,920,663 4.
evilpritted to tryiness at a temperature below 30°C 3. The precipitate of chanoclavinc diaccitatic (raw) is
under subat mospherical pressures. The residue is separated by filtration upon repeated washings with
formed by the alkaloid fraction of Kaladana, that is po water and then purified.
in thea Calonyction (Choisy) Hallier f. . nova species and, The lysergol obtaincd according to the present inven
when examined in thin-layer chromatography, hy using 5
tion, it has becn found, exhibits intc.csting phsiological
as thc solvent substance mc thylene chloridc-mic thanol. properties which manifest themsclves on thc test ani.
benzenc (25:5:5) and as the detcctor a 3% solution in mals on different systems and especially on the ncrvous
alcohol of vanillin and 0.5% by volumc of concentrated system and thc circulatory system. The substance, in
sulphuric acid, after activation at 10°C - 120°C during very low dosages, acts as a hypot.cnsive which is partic.
5 minutes, exhibits thc presence of scveral crgulic alka ()
ularly active its to its effect on the periphcral circula.
loids, limong which lysergol predominates, and, se tory system. Thc substance exhibits considerable psy.
condly, chanoclavine. chotropic properties with antiserotonin effects. The
it has bc.cn found that lysergol can he isolated by thc substanec exhibits pronounccu analgesic properties.
total alkaloids by merely washing thic mass with a low Lastly, the stimulating effect on isolated organs, such as
molecular weight aliphatic alcohol such as methanol, ut crus and intestic, is considerable. Lyscrgol. () in ac
ind. likes up the resite which is insoluble in the coll, count if these features can finil a useful lic; it in in
Such a residuc. upon filtration, can be further purifiel the thrietti's fir lifferent in; thinlogical sit at it is.
by crystallization from an appropriate organic solvent. TY this in it (vull his firin it; tur in in rurite
or also from a mixture of dimethylsulphoxide and wa 2) minner, as such. Yr in the firin if still ille silts with :-
ter. l.ysergol is especially well purificd by repeated propriac excipients suitable for both parenthcral and
crystallizations from dinncthylsulphoxide-watcr in thc oral use.
ratio of 1:1; the product thus obtaincd exhibits the fol Examples are now given by way of examplc only and
lowing properties: the percentage analysis corresponds without any linitation.
to the raw formula CHON.
C% Calcd, 75.55; H% Calcd. 7.4 N. Calcd. 11.02. 25 EXAMP.E
Found: 75.39 Found: 7.22; Found: 0.96. 40 kgs. of Kaladana secds arc ground in a mill so as
mol.wt. 2543; m.p. (crystallised from alcohol).253 to obtain a flour having a size of from 40 to 60 meshcs
°C-255°C. (dccomp.). (a) = +54 (c-0.3 in pyri per cnn.
cline).
According to another aspect, the present invention 30 The ground drug is exhausted in the cold condition in
an extrictor equipped with a stirrer, five times with gas
provides for the further isolation of chanclavine frin line having a biling point of 80'C - 90°C. 150 liters of
the residuc which contains the total crgolic alkaloids, solvent are cmployed each time. Every extraction step
upon isolation of lysergol. To this end the present in. takes three hours with stirring. The lefat text trug, up in
vention provides for the further steps of: 35 comple turcinoval of the gastline, is extracted in the
a. evaporation to dryncss, in a vacuo and at a temper same apparatus twice with a mixture of chloroform
ature below 60°C, of thc mother liquors which arc left methanol-immoria in the ratios 9:().9:0.1. 160 liters of
after the extraction of lysergol; the mixture arc employed.
h. (lissolution of the residue in an excess of pyridine Subsequently, the mass is extracted three additional
in ind clition facetic acid anhydrile in an annotint hy () times with chloroform only ( 100 liters).
weight equal to that of the dried residue, allowing the . The con hind extracts are evaporated under subi t
mixture to stand during 24 hours; mospherical pressures at a temperature below 30°C
c, precipitation of thc chanoclavine diacetate by until obtaining a volune of 25 liters,
pouring the reaction product obtainca in an excess of The concentrate is allowed to stand in a refrigerator
water an ice, incl 45 it ('C' - 4"(. ltring two lays. The solid which has been
d. purification of the chanoclavine diacciatic (raw) separated is collected by filtration with a suction pump
a?ter drying by recrystallization from ethyl acctate. and the cake is dric under witcuum. The dricd cakc
Thc thusly obtained chanoclavine diacctate has the (300 to 400 grams) is slurrica cold in 10 times its
following properties: . .
mciting point 72°C-173°C - (a)-534 (c-0.95 so iswcight of watcrand stirred during onc hour. Filtration
repeated and thc watcr insoluble residuc is dissolved
in pyridine): (a) -58 (ca 1% in chloroform). in thrice its weight of methanol; thc mc thanol solution
I.R: spectrum in CHCl characteristic absorption bands being added to the main filtrate. The filtratic is now
at 625 cm (N-COCH); 730 cm (O-COCH) washcd with 6 liters of water in a separator. The or
- U.V. spectrum in McOH + 1% CHCl: A max 283 ganic phase is theti further evaporated until a volume of
nm ( lip 3.81); 2 i? nm (log e 4.76); 29t inn (log e 55 cight liters is attained, in a vactic and at a temperature
78), .
below 30'('. This concentrate is exhausted by extract.
Analysis for CHON, (340.4) ing it four times with a 5% solution of phosphoric acid.
Caled. C# 70.6; H% 7. I; N% 8.2. There is employed, in total, eight liters of thc acidic so.
Found: 70.3; 7.5; 8,3. lution. Thc combined acidic extracts are made slighly
As regards the stages of
it should hc recalcd that:
the mcthod outlined above, 60 alkaline with aqueous anmonia until reaching a pH of
8 and are exhaustcd by extracting them with a chlorch
. Thc dry residuc as obtainca upon cvaporation of furn-mchantil mixture in thc ratio 7:3, Four extric.
the muther liquors combined of the lyscrgol cxtraction, tions are carried out by enploying, as a total, 8 liters of
is weighed and dissolved, pre?criably in about three thc mixture.
times its weight of anhydrous pyridine; e Thc organic extracts are washed once with water,
2. The excess of watcr and ice for the precipitation of then they are dried over anhydrous sodium sulphate
chanoclavinc diacetatc is about ten times the volume of and evaporated to dryncss, under subatmosphcrical
thc reaction product of the stage(b); pressures at a temperature below 30°C.
 S 3,920,663
The residuc (200 to 270 grans) as formed by the 6
total ergolic alkaloids of the drug has thc composition 5. A method according to claim 3, whicrein thc ex.
indicated in thc introductory part herco? and is now tricts are cumbinct and concentrated to one tenth of
stirred in the cold with 500 grams of mcthanol during their original volume at a temperature below 30°C
hour. The insoluble portion is collcctcd on a filtcr and s unulcr sub-atmospheric pressure, arc allowed to stand a
dried in a vacuo. few days at 0°C - 4°C and subscquently filtercd, thc fil
The further purification of the insoluble fraction tratic is washcd with watcr, thc filtrate is made alkalinic,
(lyscrgol) is carried out by dissolving the raw product and is extracted with an aqueous acidic solution, to scp
in an equal amount by weight of dimethylsulphoxide. a rate the ergotic alkaloids from the combined acidic
ly heating gently on a water bath. The solution is () extracts, said latter extraction bcing continued until the
treated with decolorizing charcoal, filtcred carcifully Ehrlich alkaloid test is negative, the alkaloids are ex
and the filtrate is supplcment.cd with an cqual volume tracticd tigain with a mixture of a chlorinated aliphatic
of distilled watcr, Then it is allowed to stand for crystal. hydrocarbon and an aliphatic alcohol, thc ncw cxtrac.
lizing. The crystallizatc is scparatcd by filtration and tion being continued until said Ehrlich tcst is negative
dried in a vacuo to constant weight. 15 again, and thc combinca extracts, washcd once only
The opcration is repeated until obtaining a product with watcr and dried ovcr anhydrous sodium sulphatic
having the properties as indicated in the introduction are evaporated to dryncss at a temperature below 30°C
herconf. untler subatuspheric pressure, furning : resilic
composed if the alkalid fraction if the extracted
EXAMPLE 2 lant, in which lysergol reliminates.
Thc nic thanolic mothcr liquors as obtained from thc 6. A mc thod according to claim 5, wherein said aque
washing of the ergolic total alkaloids of Examplc l are ous acidic solution is a solution of phosphoric acid.
placcd in a rotary evaporator and the solvent is contin 7. A mcthod according to claim 5, wherein said mix
uously removcd in a vacuo to dryness at a temperaturc ture for the sccond extraction stage is a mixture of
below 60°C. The rcsiduc (70-90 grams) is takcn up 25 chloroform and mc thanol in the respective volumc
with anhydrous pyridinc (210 - 270 mls.) and the solu ratio of 7:3.
tion
mils.).is supplcmcnted with acetic acid anhydride (70-90 8. A method according to claim 5, wherein the acidic
extracts are made alkalinc. with aqueous ammonia be.
Thc mass, sheltercd from moist air, is allowed to forc thc second extraction stage.
stand during 24 hours. Subsequently is poured over 30 9. A mcthod according to claim 5, whercin lyscrgol is
watcr and ice (2000 - 2200 mls.). The product which isolated from said residuc by washing with a low molec
scparatics is allowed to stand and is repeatedly washcd ular weight aliphatic alcohol, such as methanol, fil
with water. tered, and further purifical by crystallization with a sol
it is thcnfiltcred under vacumn, dricd and recrystal vent selected from the group consisting of organic sol
lizcd from cthyl accitatc. (30 - 45 grs.). Thc thusly ob 35 vents
taincd chanoclavinc diaccitatic is pure when cxamined a 1:1 and ratio.a mixture of dimethylsulphoxide and watcr in
in thin layer chromatography on a Kicsclgcl GF layer 10. A method for thc cxtraction of chanoclavine.
(Type 60) by using methylcne chloridc-benzcnc wherein the mother liquors, which arc left from the
methanol 25/515 as the solvent and vanillin and sul lyscrgol isolation stcp as claimed in claim 7, are evapo
phuric acid as thc reagent. 40 rated to dryness under vacuum and at a temperature
What is claimed is: P
1. A method for thc preparation of lyscrgol and er cess below 60'C, thc dry residue being dissolved in an ex
golic alkaloids comprising the stcps of extracting thesc addedofthcreto anhydrous pyridinc. and acctic acid anhydride is
alkaloids with a solvent from thc sccds of the plant Kal the residue, thcinreaction an amount by weight equal to that of
mixturc being allowcd to stand
adana, that is, a plant of the family Convolvulaccac, during 24 hours, the chanoclavinc
section of Ipomocac, genus Calonyction (Choisy) Hal cipitated by pouring the mixture asdiaccitate obtained
being pre
from the
licr f... nova specics, and concentrating the extract.
2. A method according to claim 1, whcrcin the secds clavine diacetate being purifica after drying it, chano
reaction in an excess of water and ice, the raw
by re
of thc plant arc ground to a flour and fat-stripped and
then extractcd with a halogen-substituted aliphatic hy SO crystallization from ethyl acetatc.
drocarbonaccous solvent sclectcd from the group con ccss 11. A method according to claim 0, wherein the ex
sisting of chloroform, carbon tetrachloride, and methy weight of anhydrous pyridine is about three times thc
lenc chloride, and at a temperature from 10°C to 50°C. of the dry residue.
the dix traction step being rcreatcc from 3 to 5 timcs. 12. A method according to claim 10, wherein the ex
3. A method accortling to claim 2, wherein the Not 55 cess of watcrproduct. and ice is about ten times the volume of
vent is supplemented by an amount, not exceeding t5% the retion
by volume with respcct to thc solvent, of a low molecu tion .ofAthe method certing to claim 1 wherein a frac.
plant is dissolved in a halogen substituted
lar weight aliphatic alcohol selected from the group
consisting of mc thanol, ethanol and isopropanol, and aliphatic hydrocarbon solvent, the extract is evapo
rated in filtercd, the filtrate is then extracted with an
an alkali metal in an amount, not excccding 5% by vol 6) aqueous acidic solution, thc acidic cxtract is then made
umc with respcct to thc solvcnt.
4. A nethod according to claim 2, where in the kil extracted slightly alkaline, and thc alkaloids so formed arc again
vent is supplemented by a liquid sclected from the drupcir honwith a mixture upf an aliphatic chlorina cd hy.
group consisting of mc thanol, ethanol and isopropanol and dried. an aliphatic alcohol, and then wushcd
and
and annonium hydroxidc. g s t