PHYTOTHERAPY RESEARCH, VOL.

12, 346–349 (1998)

Analytical Study and Analgesic Activity of

Oripavine from Papaver somniferum L.

M. P. Gómez-Serranillos,* O. M. Palomino, E. Carretero and A. Villar

Departamento de Farmacologı́a, Facultad de Farmacia, Universidad Complutense de Madrid, 28040, Madrid, Spain

After the isolation of an alkaloid from Papaver somniferum L. var. nigrum, its structure was determined
by means of IR, UV and 1H and 13C-NMR spectroscopy, and MS, together with a selective O-methylation
assay. The alkaloid was identified as 6,7,8,14-tetrahydro-4,5-epoxy-6-methoxy-17-methyl morphinan-3-ol
(oripavine). The analgesic activity of this compound was studied in the mouse using the hot plate techni-
que after i.p. drug administration. The effect lasted 60–90 min. The percentage analgesic effect (% A)
and lethal doses (LD50) are presented. Oripavine appears to have a similar analgesic potency to mor-
phine, but a lower therapeutic index because of severe toxicity. Toxic signs of oripavine were clonic-tonic
convulsions followed by death. # 1998 John Wiley & Sons, Ltd.
Phytother. Res. 12, 346–349 (1998)

Keywords: Papaver somniferum; phenantrenic alkaloids; HPLC; oripavine; analgesic activity.

INTRODUCTION
Phytochemical studies of plants of the family Papaver-
aceae have been directed primarily towards the alkaloids
that have been intensively investigated during the past 25
years (Grundon, 1983; Santavy, 1979, Parker et al., 1972;
Phillipson et al., 1976). Oripavine has been detected in
five different chemiotypes from Papaver orientale L.
(Section Macrantha); in chemiotype A it is the only main
alkaloid, while chemiotypes B, C, D and E have other
main alkaloids apart from this compound (Shafiee et al.,
1977). Oripavine, together with thebaine, was isolated as
the main alkaloid from some hybrids of P. bracteatum
and P. orientale, while isothebaine and traces of
oripavine were obtained from hybrids of P. orientale
and P. pseudo-orientale (Sariyar and Phillipson, 1977;
Shafiee et al., 1975; Milo et al., 1988; Nielsen et al.,
1983; Kuppers et al., 1976; Meshulam and Lavie, 1980;
Theuns et al., 1977). It can be concluded, on the available
evidence, that the major alkaloid of P. bracteatum is
thebaine, whereas that of P. orientale is oripavine. The
section Papaver is probably the most heterogeneous of
the genus Papaver according to its chemical composi-
tion. The presence of the morphinic alkaloids thebaine,
codeine and morphine together with narceine, nornar-
ceine, narcotine and narcotoline is characteristic of
Papaver somniferum and Papaver setigerum DC (Kirby
et al., 1972). The study of these alkaloids is important as
they constitute a source of medicinal compounds
including morphine, codeine, thebaine, noscapine and
papaverine.
Nowadays, morphine is still the most potent analgesic
for the treatment of strong pain and codeine is used as an
* Correspondence to: M. P. Gómez-Serranillos, Departamento de Farma-
cologı́a, Facultad de Farmacia, Universidad Complutense de Madrid, 28040,
Madrid, Spain.
antitussive. The search for new compounds that have the
same analgesic properties as morphine is due to the
necessity of eliminating the undesirable effects of this
alkaloid. Oripavine analogues and derivates such as
heterocodeine (Eddy, 1935) and etorphine (Blane et al.,
1967), have been found to be potent analgesics. Never-
theless, little information is available about the pharma-
cology and toxicity of oripavine.
This paper is part of our ongoing studies on the
chemical and pharmacological properties of the phenan-
trenic alkaloids from Papaver somniferum L. var.
nigrum. After the study of its alkaloids content, we
identified another alkaloid that was characterized as
oripavine. Once this compound was isolated, we tested its
analgesic activity and toxicity in mice in comparison with
morphine and thebaine activities.
MATERIALS AND METHODS
Plant material. The poppy straw (the capsule and the
superior 10 cm of the stalk) was supplied by Alcaliber,
S.A., Toledo, Spain. The extraction was done as
described previously (Gómez-Serranillos et al., 1994).
Pure samples of morphine and thebaine were supplied by
the Ministry of Health. The analytical HPLC system was
as reported previously (Gómez-Serranillos et al., 1994).
The semipreparative Hypersil C18 column (5 mm,
25 cm  7 mm) was supplied by Shandon Stfc. Ltd.
(UK) and runs were performed under the same analytical
conditions. The flowrate was 3 mL/min and samples of
500 mL were injected. To check the peak purity, the
chromatographic separations were monitored by a
photodiode array detector (Varian Polychrom 9065).
The acquisition of the UV spectra was automatic (l 190–
400 nm). The three spectra corresponding to the upslope,
apex and downslope of each peak were normalized and

CCC 0951–418X/98/050346–04 $17.50

# 1998 John Wiley & Sons, Ltd. Accepted 16 March 1998
 ANALGESIC ACTIVITY OF ORIPAVINE
the plots superimposed. Peaks were considered pure
when there was exact coincidence between the three
spectra (match factor  990).
Isolation of oripavine. Several aliquots of the purified
sample solution were run on the semipreparative column
under the analytical conditions described above. The
eluted fractions were collected individually and the
solvent evaporated to dryness under reduced pressure.
Individual components were tested for purity on
analytical HPLC.
Physicochemical assays and characterization of or-
ipavine. Solubility was determined as indicated in the
USP (1975). The fusion point was evaluated in an open
capillary in a Büchi-Tottoli apparatus. Rotatory power
was determined in an AA-2 polarimeter (Optical
Activity, Ltd) after dissolving the compound in chloro-
form and methanol (0.91 and 1.20 p/v, respectively). IR
spectra were run as KBr discs on a Perkin-Elmer model
577 spectrophotometer and UV spectra were obtained
with a Beckman DU 40 in methanol. 1H-NMR and 13C-
NMR spectra were recorded on a Brucker AM 200
instrument operating at 200 MHz, and chemical shifts
values are expressed as ppm (d) with TMS as an internal
standard. MS were determined with a Hewlett-Packard
model 5995 spectrometer. The temperature of the ion
source was 200 °C and the accelerating and ionizing
potentials were 3 KV and 70 eV, respectively.
Trimethylphenylammonium trioxide was used for the
O-methylation assay as the methylating agent. Thebaine
obtained in this experiment was isolated and precipitated
as thebaine bitartrate. This assay was checked and
followed by HPLC.
Toxicity and analgesic activity
Methods. Male mice NMR (Iffa-Credo Laboratory,
L’Arbresle, Fr.) 20–23 g weight were used for testing
the analgesic effects. Ten animals per group were housed
in air conditioned quarters within a 12–12 h light-dark
cycle. All animals were kept for at least 4 days after
arrival before being used in an experiment. Each animal
was housed individually when an experiment was
conducted and each mouse was used only once.
Toxicity of oripavine. The toxicity of oripavine in mice
was studied by the i.p. injection of increasing doses of the
drug. The volume of injection was 0.2 mL/mouse. The
death of the animals was recorded at 0.5, 1, 2, 4 and 24 h
after the drug administration. The mean lethal dose of the
drug (LD50), was calculated according to Miller and
Tainter (1984).
Irwin’s multidimensional screening. Mice were placed
in single cages, and observed according to standardized
worksheets at 15, 30, 60, 120 and 240 min after the
material administration. The assays were performed
using intact conscious NMR mice (weight range 20–
22 g) following the experimental conditions described by
Irwin.
Measurement of analgesic activity. The analgesic
activity of oripavine, morphine and thebaine in the
mouse after i.p. administration of the drug was measured
# 1998 John Wiley & Sons, Ltd.
347
by hot-plate method (Yeh and Mitchell, 1971) model
Socrel DS 37 (T 53.5 °C). Each subject was its own
control; thus, before treatment, its reaction time was
determined twice in 10 min, the mean of these two values
constituting the reaction time before treatment (Tb).
Thirty minutes after treatment, the reaction time was
again evaluated, but only once, this value representing
the reaction time after treatment (Ta). For each group, the
average reactions times (T) were calculated, and the
variation percentages expressed by the following ratio:
(Ta - Tb)  100/Tb.
The compound was tested at 6.5, 8.7 and 13 mg/kg
(1/4, 1/3 and 1/2 of LD50, respectively). The volume of
drug injection was 0.2 mL/mouse. Morphine and the-
baine were both used at 10 mg/kg. Analgesic activity was
considered to be present at any measurement following
drug administration when the reaction time was greater
than that of the mean control (before drug administration)
plus two standard deviations.
Drug and statistical analysis. Student’s two-tailed t-test
was used for the comparison of two experimental groups
with values of p < 0.05 considered to be statistically
significant.
Oripavine hydrochloride was dissolved in saline.
Morphine and thebaine hydrochloride were obtained
from the Ministry of Health. All drug concentrations are
expressed as free base. All used drugs were assayed by
HPLC and shown to be 99.8% pure.
RESULTS
Oripavine
The compound was an amorphous white powder. MP
201°–204 °C. The UV, IR optical rotation, 1H-NMR and
13
C-NMR were identical to previously published spectra
(Shafiee et al., 1977). The structure of oripavine (Fig. 1)
was confirmed by methylation to produce thebaine as
determined by HPLC.
Toxicity of oripavine and analgesic activity
Between 5–10 min after i.p. injection of oripavine at a
concentration greater than 25 mg/kg, mice showed clonic-
Figure 1. Oripavine structure
Phytother. Res. 12, 346–349 (1998)
348 M. P. GÓMEZ-SERRANILLOS ET AL.

Table 1. Lethality of oripavine in mice

Dose (mg/kg i.p.) Number of dead/Number of tested Lethality percentage Ratioa
10 0/10 0 0/29
20 0/1 0 0/19
25 6/10 40.00 0/15
30 8/10 73.68 14/19
35 9/10 88.46 23/26
40 8/10 93.93 31/33
80 10/10 100 41/41
LD50 (mg/kg): 26.1
a
Ratio between the number of accumulated dead animals and the number of total animals.

Table 2. Effects of morphine, thebaine and oripavine (1, 2 and 3) on pain threshold of mice in hot plate test (morphine and
thebaine doses, 10 mg/kg; 1, oripavine 6 mg/kg; 2, oripavine 8.7 mg/kg; 3, oripavine 13 mg/kg, i.p.)
Reaction time (s)
0 min 30 min 60 min 90 min 180 min
Sample na mean  SD mean  SD mean  SD mean  SD mean  SD
Morphine 10 10.5  1.3 19.7  2.3 18.5  1.9 15.2  1.2 13.8  1.1
Thebaine 10 5.9  1.7 8.6  2.1 9.7  2.4 9.1  1.7 7.6  1.6
1 10 8.8  1.7 15.3  3.0 14.9  3.6 12.4  2.3 10.4  1.7
2 10 9.7  1.4 16.3  2.7 13.2  2.8 10.9  1.9 12.7  1.9
3 10 9.5  1.7 15.9  3.7 13.8  3.0 13.3  3.8 10.7  1.9
p < 0.05 compared with control group (n = 10)
a
Number of mice per group.

tonic convulsions, muscle-rigidity and, eventually, death.
The animals died within 2–3 min when large doses of
oripavine were employed. When the animals survived for
60 min, they lasted more than 4 days (Table 1).
Irwin’s test
In mice, all oripavine dosages assayed (6.5, 8.7 and
13 mg/kg) produced a decrease of the alert status,
spontaneous activity and reactivity. Oripavine also
caused a central nervous system excitation that showed
as a sacrococcigeo dorsal muscle contraction (Strauss
reaction); no motor incoordination was observed. A
decrease in the respiratory rhythm was observed after the
administration of all three doses.
Analgesic activity
The oripavine analgesic effects in mouse were observed
with a peak effect at 30 min and lasted about 90 min after
the drug administration. At all three doses, the compound
showed significant analgesic action in the hot-plate
method. It is known that centrally acting analgesic drugs
can elevate the pain threshold of mice towards heat and
pressure. From the above findings it is clear that
oripavine raised the pain threshold, which indicates that
is centrally acting.
DISCUSSION
The analgesic potency of oripavine was comparable to
# 1998 John Wiley & Sons, Ltd.
that of morphine and thebaine. When the doses were
larger than 15 mg/kg, convulsions were observed in some
animals. The analgesic effects of oripavine, morphine
and codeine are shown in Table 2. The results obtained
after the quantification of oripavine in P. somniferum
(Gómez-Serranillos et al., 1994; Gómez-Serranillos et
al., 1995) together with the previous analgesic studies
(Villar et al., 1992) justified the characterization
(physicochemical assays) and the pharmacological study
of this alkaloid. Its structure (free phenolic hydroxyl
group as occurs in morphine) led us to suspect an
equivalent analgesic activity to morphine.
From the pharmacological point of view, the intraper-
itoneal administration of oripavine, showed a high acute
toxicity in the mouse (LD50 26.2 mg/kg) which was much
higher than morphine (LD50 500 mg/kg). Concerning the
effects of oripavine on behavioural parameters, the three
assayed doses also produced an excitant effect. The
analgesic potency of oripavine appeared to be similar to
that of morphine in the mouse. Since the structure of
oripavine is similar to that of thebaine, a comparison
between the analgesic activity of these two compounds
became interesting; in this way, both compounds showed
a similar activity profile with a maximum at 30 min;
nevertheless, oripavine activity was much higher than
thebaine. The thermic analgesic test employed (hot-plate
test), as well as the response to pain, implies the
participation of m and d receptors in the central nervous
system at both spinal and supraspinal levels.
In conclusion, these results show that oripavine from
Papaver somniferum L. (poppy straw) presents important
excitant effects and analgesic properties.
With the aim of explaining the mechanism of action of
this product as an analgesic agent, further investigations
will be carried out in our laboratory.
Phytother. Res. 12, 346–349 (1998)
 ANALGESIC ACTIVITY OF ORIPAVINE
REFERENCES
Barton, D. H. R., and Cohen, T. (1957). Some biogenetic
aspects of phenol oxidation. Festschr Stoll. BirkhaÈuser
Basel, 2, 117±143.
Blane, G. F., Bourr, A. L. A., Fitzgerald, A. E., and Lister, R. E.
(1967). Actions of etorphine hydrochloride, (M99): A
potent morphine-like agent. Br. J. Pharmacol. Chemo-
ther. 30, 11±22.
Eddy, N. B. (1935). Pharmacology of metopon and other new
analgesic opium derivatives. J. Pharmacol. Exp. Ther., 55,
127±135.
GoÂmez-Serranillos, P., Carretero, E., and Villar, A. (1994).
Analysis of poppy straw and poppy straw concentrate by
reversed-phase high-performance liquid chromatograpy.
Phytochem. Anal. 5, 15±18.
GoÂmez-Serranillos, P., Carretero, E., and Villar, A. (1995). A
new method for the analysis of alkaloids in poppy straw.
Fitoterapia LXVI, 156±158.
Grundon, M. F. (1983). The Alkaloids, pp. 6±13. Chemical
Society London.
Kirby, G. W., Massey, S. R., and Steinreich, P. (1972).
Biosynthesis of unnatural morphine derivatives in P.
somniferum. J. Chem. Soc., Pekin Trans I, 1642±1647.
Kuppers, F. J. E. M., Salemink, C. A., Bastart, M., and Paris, M.
(1976). Alkaloids of Papaver bracteatum: presence of
codeine, neopine and alpinine. Phytochemistry, 15, 444±
445.
Meshulam, H., and Lavie, D. (1980). The alkaloidal constitu-
ents of P. bracteatum Arya II. Phytochemistry, 19, 2633±
2635.
Miller, L. C., and Tainter, M. L. (1984). Estimation of the DE50
and its error by means of logarithmic-probit graph paper.
Proc. Soc. Exp. Biol. Med. 57, 261±264.
Milo, J., Levy, A., Palevitch, D., and Ladizinsky, G. (1988).
High-performance liquid-chromatography analysis of the
alkaloid spectrum in the roots and capsules of the species
# 1998 John Wiley & Sons, Ltd.
349
and hybrids of Papaver section oxytona. J. Chromatogr.
452, 563±570.
Nielsen, B., Roe, J., and Brochmann-Hanssen, E. (1983).
OripavineÐa new alkaloid. Planta Med., 48, 205±206.
Parker, H. I., Blaschke, G., and Rapoport, H. (1972). Biosyn-
thetic conversion of thebaine to codeine. J. Chem. Am.
Soc., 94, 1276±1282.
Phillipson, J. A., Handa, S. S., and El-Dabbas, S. W. (1976). N-
oxides of morphine, codeine and thebaine and their
occurrence in Papaver species. Phytochemistry, 19,
1681±1684.
Santavy, F. (1979). Papaveraceae alkaloids. In The Alkaloids
ed. by R.H.F. Manske, 12, pp. 333±385. Academic Press,
London, New York.
Sariyar, G., and Phillipson, J. D. (1977). Macrantaline and
macrantolidine, new alkaloids from Turkish sample of
Papaver pseudo-orientale. Phytochemistry 16, 2009±
2013.
Sha®ee, A., Lalezari, I., Nasseri-Nouri, P., and Asgharian, R.
(1975). Alkaloids of Papaver orientale L. and Papaver
pseudo-orientale. J. Pharm. Sci. 64, 1570±1572.
Sha®ee, A., Lalezari, I., Assadi, F., and Khala®, F. (1977).
Alkaloids of Papaver orientale L. J. Pharm. Sci. 66(7),
1050±1052.
Theuns, H. G., Dam, J. E. G., van Luteijn, J. M., and Salemink,
C. A. (1977). Alkaloids of Papaver bracteatum: 14-b
hydroxicodeinona, 14-b hydrocodeine and N-methyl
corydaldine. Phytochemistry, 16, 753±755.
USP (1975). The United States Pharmacopeia, 19th rev., pp.
360±368. Mack Publishing Co., Easton, PA.
Villar, A., Carretero, E., and GoÂmez-Serranillos, M. P. (1992).
ARS Pharmaceutica. Pharmacia Mediterranea XIX, T. II,
607±613.
Yeh, S. Y., and Mitchell, C. L. (1971). Analgesic activity and
toxixity of oripavine and f-dihydrothebaine in the mouse
and rat. J. Pharmacol. Exp. Ther. 179, 642±651.
Phytother. Res. 12, 346–349 (1998)