全 文 ：Incarvilleae Alkaloids and Antinociceptive Substances
CHI Yu-ming1＊ ,NAKAMURA Motoyuki1 ,ZHAO Xi-ying1 ,
YOSHIZAWA Toyokichi1 ,YAN Wen-mei2 ,HASHIMOTO Fumio3 ,NOHARA Toshihiro3 ,SAKURADA Shinobu4
　　　　 　　天然产物研究与开发　 　　　　　
　　　　　　　　　　 NATURAL PRODUCT RESEARCH AND DEVELOPMENT 2005　Vol.17　No.3
　
　
　
　
　　Received February 7 , 2005;Accepted March 22 , 2005
　＊Corresponding author Tel:81-293-427148;E-mail:chi-59@unimatec.
co.jp
(1.Seiwa Pharmaceutical.Ltd , Ibaraki 319-1535 , Japan;
2.Beijing University of Traditional Chinese Medicine and Pharmacy , Beijing 100029 , China;
3.Kumamoto University , Kumamoto 862-0973 , Japan;4.Tohoku College of Pharmacy , Sendai 981-0905 , Japan)
Abstract:Incarvillae sinensis LAM., a traditional Chinese medicine , has been used to treat rheumatism and to relieve pain.
From the whole plants , a number of novel monoterpene alkaloids and macrocyclic spermine alkaloids have been identified.One
of the monoterpene alkaloids , incarvillateine , demonstrated higher antinociceptive effect than morphine in the formalin test , and
the mechanism of antinociception was different from that of morphine.Incarvillateine has become an important lead compound
for developing new natural medicinal drugs.In this paper , the chemical investigation as well as the biological activity and struc-
ture-activity relationship were reviewed.
Key words:Incarvillea sinensis;monoterpene alkaloid;incarvillateine;antinociceptive effect
角蒿生物碱及镇痛活性物质
迟玉明1＊ ,中村基之1 ,赵唏瑛1 ,吉泽丰吉1 ,阎文玫2 ,桥本文雄3 ,野原稔弘3 ,樱田忍4
(1.日本正和药品株式会社 , 茨城 319-1535;2.北京中医药大学 ,北京 100029;
3.熊本大学 , 熊本 862-0973;4.东北药科大学 ,仙台 981-0905)
摘　要:角蒿(Incarvillea sinensis)为透骨草的主要来源之一 ,称为“羊角透骨草” , 具有祛风除湿 、消肿止痛之功效。
从其全草分离得到了多种新的单萜生物碱和大环精胺类生物碱 , 其中单萜生物碱之一角蒿酯碱(incarvillateine),
具有很强的镇痛活性 ,且作用机理不同于吗啡。 角蒿酯碱已成为开发新型非麻醉性镇痛新药的重要先导化合
物 ,本文对角蒿的化学成分 、镇痛活性 、作用机理和构效关系的研究作一综述。
关键词:角蒿;单萜生物碱;角蒿酯碱;镇痛活性
中图分类号:O629.3　　　
Incarvillea sinensis (Bignoniaceae) is one of the main
sources of the Chinese crude drug “Tougucao” , and it is
mainly used in the north and northeast areas of China
[ 1] .
We had designed to assign the pharmacological active
substance(s)from this plant.So far , a number of novel al-
kaloids have been identified and one of the monoterpene
alkaloids , incarvillateine , was found to possess higher
antinociceptive effect than morphine in the formalin test
with a different mechanism.
Novel Alkaloids from I .sinensis
From the whole plants of I.sinensis , seventeen novel com-
pounds with one known compound have been isolated.Ac-
cording to their structures , these compounds could be di-
vided into two groups:monoterpene alkaloids and macro-
cyclic spermine alkaloids.
The absolute configuration of incarvilline(1), a core com-
pound of monoterpene alkaloid type ,was determined by X-
ray crystallography[ 2 , 3] .Oxyincarviline (2)was 5-hydrox-
ylated incarvilline[ 3] .Incarvine A (3)[ 4] and incarvine B
(4)[ 5] were the esters of Hildebrant s acid and incar-
villine.Incarvine C (5)[ 6] was a deoxytetrahydro-deriva-
tive of incarvine B , but the incarvilline esterified by
Hildebrant s acid in a head position.Incarvine A N-oxide
(6)[ 6] , an N-oxidative product of incarvine A ,was found
to be a mixture of N-oxide and N -oxide.Incarvine D
(7)[ 5] was an ester of ferulic acid and incarvilline , and
this compound is an important intermediate of incarvil-
lateine (8).Incarvillateine had a dimeric structure of in-
carvine D , and the structure was determined by X-ray
362
DOI :10.16333/j.1001-6880.2005.03.030
analysis[ 7] .Methoxyincarvillateine (9)was 5″or 5 -
methoxylated incarvillateine[ 6] , Incarvillateine C(10)and
Incarvillateine D(11)were bis-demethoxy and demethoxy
derivatives of incarvillateine , respectively[ 8] .Incarvil-
lateine N-oxide(12)was also found to be a mixture of N-
oxide andN′-oxide of incarvillateine[ 6] .Five novel macro-
cyclic spermine alkaloids named incasines A , A′, B , B′
and C (14 ～ 18)were isolated together with a known al-
kaloid verballocine (13)[ 9] , and the absolute structures
were determined by CD spectra of their saturated prod-
ucts
[ 10] .
Fig.1　The alkaloids from Incarvillea sinensis
Antinociceptive Effect
In parallel with the chemical investigation , the antinoci-
ceptive effects have been examined by formalin test in
mice.The 60%methanol eluent(120 mg/kg , i.p.)from
Diaion HP-20 column chromatography of the MeOH ex-
tract showed significant antinociceptive effect in both early
and late phases of the formalin-induced pain response.
Further isolation of this fraction gave the active principle ,
incarvillateine.Incarvillateine was found to produce grad-
ed inhibition of both the neurogenic(early phase)and in-
flammatory (late phase)phases in a dose-related man-
ner[ 11](Fig.2).Comparing the antinociceptive effects of
different doses of incarvillateine and morphine , the ED50
values of incarvillateine were about 1.06 (early phase)
and 1.33 (late phase)times lower than those of mor-
phine[ 11](Table 1).
Incarvillateine was a representative of the novel monoter-
pene alkaloidal derivatives from I .sinensis.It presented
an unique feature of having a dimeric structure in the
molecule.
3632005　Vol.17　No.3 迟玉明等:角蒿生物碱及镇痛活性物质 　
Fig.2　The effects of incarvillateine on the early and late phases of formalin-induced(1%, 20 μL)pain response in mice.Each
value representsmean ± SE(n=10).Significant differences between control and drug treated groups are indicated by＊＊
<0.01
Table 1　Comparing effect of incarvillateine(i.p.)and mor-
phine(i.p.)on formalin- induced(1%, 20 μL)pain
response in mice(n=10)
Morphine(ED50) Incarvil lateine (ED50) I/M
Early phase 18.4μmol/ kg 17.4μmol/ kg 1.06
Late phase 10.4μmol/ kg 17.8μmol/ kg 1.33
Mechanism of Antinociception
In order to investigate the antinociceptive mechanism of
incarvillateine(INCA), some selective opioid-receptor an-
tagonists , such as nor-binaltorphimine(nor-BNI),β-funal-
trxamine (β-FNA)and naltrindole(NTI)were pretreated
prior to the injection of INCA in the formalin test[ 11] .The
antinociceptive effects of INCA in both early and late
phases were markedly reduced by s.c.pretreatment with
β-FNA , a selective μ-opioid receptor antagonist , and nor-
BNI , a selective κ-opioid receptor antagonist.On the other
hand , these antinociceptions were not antagonized by s.c.
pretreatment with NTI , a selective δ-opioid receptor antag-
onist.Furthermore , the effects of INCA could also be
blocked by theophylline (THEO), an adenosine-receptor
antagonist (Fig.3).These results suggested that the
antinociceptive effect of INCA might not only correspond
to μ- andκ-opioid receptors ,but also to adenosine-recep-
tor.Incarvillateine might become an important lead com-
pound for developing a new antinociceptive medicine hav-
ing a mechanism of action different from that of morphine.
Fig.3　Effects of incarvillateine(20 mg/ kg , i.p.)and reversed effects of β-FNA(40 mg/ kg , s.c.), NTI(0.5 mg/kg , s.c.), nor-BNI
(20 mg/ kg , s.c.)and theophylline(5 mg/ kg , s.c.)on formalin-induced(1%, 20 μL)pain response in mice.Each value rep-
resents mean±SE(n=10).Significant differences between control and drug treated groups are indicated by＊＊<0.01
Structure-Activity Relationship of Incar-
villateine
The relationship between the structure and the antinoci-
ceptive effect of incarvillateine was examined
[ 12] .The es-
sential factors for the appearance of the antinociceptive ef-
fects were shown to be the presence of a monoterpene al-
kaloid moiety , incarvilline , and a dimeric structure carry-
ing a cyclobutane ring in its component[ 12] .This sugges-
tion was supported by the result that the antinociceptive
intensity of 3 , 3′-demethoxy-4 , 4′-dehydroxyincar-
villateine , a synthesized new incarvillateine derivative ,was
parallel to that of incarvillateine.Additionally , the
diphenylcyclobutane dicarboxylic acids , such asα-truxillic
364 天然产物研究与开发　　　　　　　　　　　　　 　　　2005　Vol.17　No.3
acid and 4 ,4′-dihydroxy-α-truxillic acid , exhibited signifi-
cant antinociceptive effects in the late phase of formalin-
induced pain model in mice , suggested that the cyclobu-
tane moiety of dimeric structure might play an important
role of expressing antinociceptive action on inflammatory
reaction and the diphenylcyclobutane dicarboxylic acid
derivatives might become a new type of anti-inflammatory
drugs.
In conclusion , incarvillateine , a new monoterpene alkaloid
carrying a characteristic cyclobutane ring ,has been found
to produce more potent antinociceptive activity than mor-
phine in a formalin-induced pain response.Further inves-
tigation for incarvillateine is required to elucidate the ex-
act mechanism of action and to develop a new antinocicep-
tive medicine.
Fig.4　Antinociceptive effects of various compounds on formalin- induced(1%, 20 μL)pain response in mice.1:incarvilline , 3:in-
carvine A , 7:incarvine D , 8:incarvillateine, 19:trans-ferulic acid , 20:3 , 3′-demethoxy-4 , 4′-dehydroxyincarvillateine.Each
value representsmean ± SE(n=10).Significant differences between control and drug treated groups are indicated by＊＊
<0.01
References
1　 Chi YM , Yan WM , Li JS.Original botanical statistics and com-
mercial investigation of Chinese crude drug “ Tougucao” .Chi-
na Jounal of Chinese Materia Medica , 1990 , 15:262-265
2　 Chi YM , Yan WM , Chen DC , et al.An alkaloid from Incarvil-
lea sinensis.Phytochemistry , 1992 , 31:2930-2932
3　 Chi YM , Hashimoto F , Yan WM , et al.Monoterpene alkaloids
from Incarvillea sinensis.Absolute stereochemistry of incar-
villine and structure of a new alkaloid , hydroxyincarvilline.
Chem Pharm Bull , 1995 , 45:495-498
4　 Chi YM , Hashimoto F , Yan WM , et al.Incarvine A , a
monoterpene alkaloid from Incarvillea sinensis.Phytochemistry ,
1995 , 40:353-354
5　 Chi YM ,Hashimoto F , YanWM , et al.Two alkaloids from In-
carvillea sinensis.Phytochemistry , 1995 , 39:1485-1487
6　 Chi YM ,Hashimoto F , Yan WM , et al.Four novel monoter-
pene alkaloidal derivatives from Incarvillea sinensis.Phyto-
chemistry , 1997 , 46:763-769
7　 Chi YM , Yan WM , Li JS.An alkaloid from Incarvillea sinensis.
Phytochemistry , 1990 , 29:2376-2378
8　 Nakamura M , Chi YM , Kinjo J , et al.Two monoterpene alka-
loidal derivatives from Incarvillea sinensis.Phytochemistry ,
1999 , 51:595-597
9　 Drandarov K.Verbacine and verballocine , novel macrocyclic
spermine alkaloids from Verbascum pseudonobile (Scrophulari-
aceae).Tetrahedron Letters , 1995 , 36:617-620
10　Chi YM ,Hashimoto F , Yan WM , et al.Five novel macrocyclic
spermine alkaloids from Incarvillea sinensis.Tetrahedron Let-
ters , 1997 , 38:2713-2716
11　Nakamura M , Chi YM , Yan WM.et al.Strong antinociceptive
effect of incarvillateine , a novel monoterpene alkaloid from In-
carvillea sinensis.Journal of Natural Product , 1999 , 62:1293-
1294
12　Nakamura M , Chi YM , Yan WM.et al.Structure-antinocicep-
tive activity studies of incarvillateine , a monoterpene alkaloid
from Incarvillea sinensis.Planta Medica , 2001 , 67:114-117
3652005　Vol.17　No.3 迟玉明等:角蒿生物碱及镇痛活性物质