Alkaloids of the genus Poecilanthe (Leguminosae: Papilionoideae)

B&hem&J Mtjcs

Pergamon

andEco/ogy,Vol.23, Na 6, pp.547-663.1986 Cq&ht 0 1985 ElsevierScienceLtd Printedin GreatBritainAll rightsreserved 030%1978/96 S950+0.00

o3os-l878(86)oooso-x

Alkaloids of the Genus Poecilanthe (Leguminosae: Papilionoideae) ROLAND GREINWALD,’ PETER BACHMANN,’

GWILYM LEWIS,t LUDGER WITTE,S and FRANZ-CHRISTIAN CZYGAN”

‘Lehntuhl far Pharmazeutische Biologiq Universit8tWOrzburg, Mittlerer Dallenbergweg 64, 97082 WOrzburg, Germany; tRoyal Botanic Gardens, Kew, Richmond, SurreyTW9 3AB, U.K.; Slnstitut firr Pharmazeutische Biologie derTechnischen Universitat Braunschweig, Mendelssohnstr.1, 38023 Braunschweig, Germany

Key Word Index--Poeci/antie; Leguminosae: Papilionoideae; Dalbergieae; quinolizidine alkaloids; bipiperidyl alkaloids; chemotaxonomy. Abstract-The presence of alkaloids in the genus Poecilanthe is reported for the first time. Epilupinine, 4fl-hydroxyepilupinine, dihydrolusitanine, dihydroammodendrine, lusitanine, N-methylcytisine, cytisine, lupanine, N-formylcytisine, anagyrine, baptifoline, camoensidine and four novel alkaloids were found as major alkaloids in leaves and seeds of the genus Poecilanthe. Tashiromine, acetylepilupinine, ammodendrine, a-isolupanine, 5,6_dehydrolupanine, aphyllidine, aphylline, thermopsine, epibaptifoline, rhombifoline, tinctorine, 11 -allylcytisine, acetylbaptifoline and at least seven unknown compounds were found as minor alkaloids in the extracts studied. The co-occurence of different types of quinoliridine (bicyclic lupinine-type, lupanine-type, leontidine-type and a-pyridone-type) and bipiperidyl alkaloids appears to be a good chemotaxonomic marker for the genus. The presence of alkaloids in Poeci/anthe suggests a relationship of this genus with the tribes Sophoreae or Brongniartieae rather than with the Dalbergieae. The alkaloidal data indicate that the genus can be divided into two phytochemical groups: one group of species accumulating mainly a-pyridone alkaloids and a second group accumulating bicyclic quinolizidine alkaloids, with a-pyridones being totally absent.

Introduction The genus Poecilanthe Benth. belongs to the family Leguminosae, subfamily Papilionoideae, tribe Dalbergieae (Lavin, 1987). It comprises 10 species, which occur in tropical South America. The exact taxonomic position of this genus is not yet well established and numerous problems in assessing its relationships have been encountered by taxonomists. Poecilanthe was originally placed in the Dalbergieae by Bentham (1860). Geesink referred it to theTephrosieae (now the Millettieae) in 1981. This classification was supported by investigations on the wood anatomy, which exhibits features that are rather typical for the Millettieae and absent from Dalbergieae (Baretta-Kuipers, 1981). Also in 1981, Geesink stated that the genus is apparently transitional with the tribe Robinieae, as axillary racemes seem to occur as well as panicles. Based mainly on this characteristic the genus was transferred to the Robinieae 4 years later (Geesink, 1984). Most recently Lavin (1987) referred it back to the Dalbergieae, but he pointed out that the genus could be phylogenetically isolated. The alkaloid pattern and its variation within a given genus can be used to complement morphological data in studies of taxonomy and phylogeny (Greinwald et a/., 1989; Kinghorn et a/., 1982; Salatino and Gottlieb, 1980; van Wyk et a/., 1989). Since no report on the alkaloids of this genus was found in the literature, we consider the present study to be the first for Poecilanthe. (Received 14 September 1994; accepted 12 April 1995)

548

R. GREINWALDETAL.

Materials and Methods Plant materials. A comprehensive list of voucher specimens of all the species studied is given below, with sample numbers as in the tables. Poecilanthe arnazonica (Ducke) Ducke, sample 1 : leaves, Brazil, Zarucchi eta/. 2968 (K); sample 2: leaves, seeds, Manaus---Porto Velho, Amazona state, Brazil, Lleras et al., 19638 (K). Poecilanthe effusa (Huber) Ducke, sample 1 : leaves, Par~, Brazil, Silva & Souza 2648 (K); sample 2: seeds, territory of Amap~, Brazil, Daly et aL, 3947. Poeci/anthe falcata (Veil.) Heringer, leaves, Brazil, de Lima eta/. 2852 (K). Poeci/anthe grandif/ora Benth., leaves, Rio de Janeiro, Brazil, Kuhlmann s.n. (K). Poeci/anthe hostmannii (Benth.) Amshoff, sample 1: leaves, Surinam, Lanjouw & Lindeman 2688 (K); sample 2: leaves, French Guyana, Granville 6453 (K). Poecilanthe itapuana G. P. lewis, sample 1 : leaves, Bahia, Brazil, Ribeiro eta/. 347 (K); sample 2: leaves, Bahia, Salvador, Lewis & Andrade 2018 (K). Poeci/anthe ovalifo/ia Kleinh., leaves, Surinam, Tapanahoni-river, Gonggrijp 4132 (Herb. Boschwezen). Poeci/anthe parvif/ora Benth., sample 1 : leaves, Paraguay, Hassler 11438 (K); sample 2: leaves, seeds, Rio de Janeiro exhort., Lewis s.n. (K). Poecilanthe subcordata Benth., leaves, seeds, Bahia, Brazil, Harley et a/. 21805 (K). Poeci/anthe u/ei (Harms) Arroyo & Rudd, sample 1: leaves, seeds, Bahia, Brazil, Lewis eta/. 835

(K); sample 2: leaves, Tucano, Bahia, Brazil, Goncalves 91 (K); sample 3: seeds, Bahia, Brazil, Hage 956 (K). Extraction procedure. The dry plant material was homogenized in 0.1 N H2S04 and left standing at room temp. for 20 min. The filtrated homogenate was made alkaline with 25% ammonium hydroxide and applied to Chemelut (ICT, Frankfurt) columns (0.6 g Chemelut m1-1 extract). The alkaloids were eluted with 1 O0 ml dichloromethane and the extract was evaporated to dryness. The extract was redissolved in 100 111 MeOH before GC injection. Capillary GC The extracts were analysed by GC according to published conditions (Greinwald eta/., 1 991 ). Retention indices (RI) were calculated using co-chromatographed standard hydrocarbons. Capillary GC-MS. Measurements were performed on a J & W DB1 (0.32 mm × 30 m) column coupled directly to a quadrupole MS. Carrier gas: He; split 1:20; injector: 250°C; oven: 150-300°C at 6°C rain-l; electron energy 45 eV. Reference alkaloids. /upinine, epilupinine, sparteine, ammodendrine, lusitanine, cytisine, N-methylcytisine, lupanine, anagyrine, thermopsine and baptifoline were available to us as reference samples. All these reference samples were fully authenticated by spectroscopic methods (IR, MS, NMR). The identity of dihydrolusitanine, dihydroammodendrine and acetylepilupinine was confirmed by chemical synthesis (Bachmann, unpubl.). Alkaloid identification. The alkaloids were identified by co-chromatography with the reference alkaloids, by their specific retention indices and by their mass spectral fragmentation data. The following extracts were investigated by GC-MS: (A) P. amazonica, sample 1 : leaves; (B) P. effusa, sample 1 : leaves; (C) P. falcata, leaves; (D) P. hostmannii, sample 2: leaves; (E) P. itapuana, sample 2: leaves; (F) P. effusa, sample 2: seeds; (G) P. parviflora, sample 2: seeds; (H) P. subcordata, seeds. Piperidyl alkaloids. Ammodendrine (A, C, D, E, F, G, H), RI: 1865; MS: m/z 208 (51) [M] ÷, 43 (100) (Fitch and Djerassi, 1974; Wink and Witte, 1987). Dihydroammodendrine (A, D), RI: 1850; MS: rn/z 210 (2) [M] +, 84 (100). Bicyclic quinolizidine alkaloids. Tashiromine (A, C, D, G), RI: 1280; MS: m/z 155 (61) [M] ÷, 1 54 (62), 138 (74), 124 (60), 110 (18), 97 (53), 96 (100), 84 (44), 83 (45), 69 (40), 55 (16) (Ohmiya et aL, 1990). Epilupinine (A, B, C, D, E, G, H), RI: 1415; MS: m/z 169 (43) [M] +, 168 (34), 152 (63), 1 38 (56), 124 (25), 110 (38), 98 (37), 97 (65), 83 (100), 55 (69) (Asres et al., 1986). Acetyl-epilupinine (A), RI: 1513; MS: m/z 211 (9) [M] +, 168 (2), 152 (100), 97 (28), 83 (21). 4~-Hydroxyepilupinine (A, C, D), RI: 1605; MS: m/z 185 (38) [M] +, 168 (73), 154 (27), 136 (13), 110 (28), 98 (46), 97 (100), 83 (55), 69 (10), 55 (23) (Veen et al., 1991 ). Dihydrolusitanine (A, D), RI: 1780; MS: m/z 210 (10) [M] +, 195 (1), 181 (4), 152 (22), 138 (100), 124 (12), 110 (41), 97 (29), 83 (90), 55 (39). Lusitanine (A, B, D, E), RI: 1880; MS: m/z 208 (53) [M] +, 179 (37), 166 (100), 136 (98), 123 (26), 110 (78), 82 (31), 43 (82) (Hatfield et al., 1985; Steinegger et aL, 1965). Sparteine- and lupanine-type quinolizidine alkaloids. Aphyllidine (D), RI: 2127; MS: m/z 246 (64) [M] + , 245 (33), 218 (9), 203 (6), 189 (11 ), 175 (4), 163 (17), 148 (10), 134 (17), 120 (6), 98 (100), 97 (56), 96 (30), 84 (8) (Schumann et al., 1968). c~-Isolupanine (E, G, I), RI: 2100; MS: m/z 248 (57) [M] +, 136 (100) (Wink et al., 1983). 5,6-Dehydrolupanine (E, G, H), RI: 2133; MS: m/z 246 (34) [M] +, 98 (100) (Wink et aL, 1983). Lupanine (A, D, E, F, G), RI: 2160; MS: m/z 248 (47) [M] +, 136 (100) (Wink et al., 1983). Aphylline (D, E), RI: 2180; MS: m / z 2 4 8 (35) [M] +, 247 (34), 220 (23), 205 (5), 191 (20), 149 (18), 137 (47), 136 (100), 123 (23), 110 (22), 97 (34), 84 (22), 68 (11 ), 55 (18) (Wink et al., 1983). Leontidine-type alkaloids. Camoensidine (E, G, H), RI: 2080; MS: m/z 234 (42) [M] +, 233 (37), 219 (3), 205 (3), 149 (14), 136 (55), 135 (54), 122 (100), 110 (18), 96 (37), 84 (46) (Ohmiya et aL, 1991 ). c~-Pyridone alkaloids. N-methylcytisine (C, E, G, H), RI: 1950; MS: m/z 204 (24) [M] +, 58 (100) (Wink et aL, 1983). Cytisine (C, E, G, H), RI: 1990; MS: m/z 190 (67) [M] +, 146 (100) (Wink et aL, 1983). Rhombifoline (H), RI: 2155; MS: m/z 244 (1), 203 (100) (Wink et al., 1983). Tinctorine (E), RI: 2235; MS: m/z 244 (1), 203 (100) (Wink et aL, 1983). 11 -Allylcytisine (E), RI: 2245; MS: m/z 230 (4), 189 (100) (Wink et aL, 1983). Thermopsine (C, E), RI: 2305; MS: m/z 244 (35) [M] +, 160 (13), 146 (15), 136 (14), 122 (12), 98 (100), 84 (16) (Wink etal., 1983). N-formylcytisine (C, E, G, H), RI: 2315; MS:

ALKALOIDSOF GENUSPOECILANTHE

549

m/z 218 (54) [M]+, 146 (100) (Wink et al., 1983). N-acetylcytisine (C, E, G, H), R/: 2320; MS: m/z 232 (58) [M]+, 146 (100) (Ohmiya etal., 1974). Anagyrine (E, G, H), R/: 2380; MS: m / z 2 4 4 (43) [M]+, 98 (100) ((Wink et al., 1983). Baptifoline (C, E, G, H), R/: 2635; MS: m/z 260 (32) [M]+, 114 (100) (Wink

et al., 1983). Epibaptifoline (E, G, H), RI: 2650; MS: m/z 260 (11) [M]+, 114 (100) (Greinwald et al., 1990). Unknown alkaloid~ X1 (piperidyl alkaloid ?) (D), RI: 1765; MS: m/z 196 (3) [M]+, 126 (7), 112 (8), 96 (8), 83 (13), 70 (100), 43 (36). X2 (c¢-pyridonealkaloid 7) (C, H), RI: 1890; MS: m/z 204 (23) [M] ÷, 174 (1), 160 (19), 146 (9), 132 (7), 118 (6), 106 (3), 95 (7), 82 (50), 58 (46), 57 (100). X3 (alkaloid with a lupinine skeleton ?) (B), RI: 2020; MS: m/z 250 (6) [M]+, 249 (7), 235 (1), 221 (3), 209 (2), 195 (7), 181 (10), 168 (3), 152 (37), 138 (100), 124 (26), 110 (31), 98 (42), 96 (47), 84 (48), 83 (60), 69 (12), 55 (41). X4 (E, G), RI: 2040; MS: m/z220 (31) [M] +, 219 (29), 205 (2), 192 (13), 177 (3), 163 (6), 150 (8), 136 (11 ), 122 (100), 110 (19), 96 (23), 84 (33), 70 (22). X5 (Camoensidine isomer?) (E), RI: 2098, MS: m/z234 (18) [M] +, 233 (27), 206 (26), 191 (8), 177 (17), 164 (4), 149 (12), 134 (8), 123 (74), 122 (100), 110 (24), 96 (13), 84 (57), 83 (33), 67 (9), 55 (23). X6 (D), RI: 2220; MS: m/z 278 (23), 98 (100), 96 (54), 84 (20). X7 (E), RI: 2220; 276 (27) [M] +, 259 (6), 245 (22), 231 (7), 218 (44), 203 (7), 136 (72), 135 (100), 134 (65), 127 (21), 110 (19), 96 (17), 83 (27). X8 (D), RI: 2273; MS: m/z 262 (72), 245 (16), 233 (5), 219 (2), 205 (3), 191 (3), 179 (8), 154 (8), 150 (5), 136 (23), 134 (11), 122 (13), 110 (10), 98 (100), 97 (56), 96 (29), 84 (20). X9 (Hydroxyaphylline ?) (D), RI: 2425; MS: m/z 264 (45) [M] +, 247 (9), 236 (19), 219 (4), 136 (100), 122 (17), 110 (12), 97 (52). X10 (B, F),/71. 2655; MS: m/z 327 (14) [M] +, 287 (20), 286 (100), 151 (17), 134 (5), 120 (6), 109 (20), 94 (7), 82 (8). X11 (B, F), RI: 2810; MS: m/z 341 (18) [M] +, 311 (21), 300 (100), 257 (17), 203 (5), 190 (10), 175 (3), 148 (7), 138 (9), 122 (12), 96 (11), 84 (10). Results

In leaf and seed extracts of the genus Poecilanthe, we identified 25 alkaloids belonging to the quinolizidine and biperidyl types. The alkaloids dihydrolusitanine and dihydroammodendrine were detected as new plant constituents. Both were identified by comparison of their mass spectra with those of synthetic reference samples and by co-chromatography with synthetic reference samples. The mass spectrum of dihydrolusitanine additionally agrees well with the mass spectral data given for the synthetic acetamidomethylquinolizidine (= dihydrolusitanine) (Saito et aL, 1987). The molecular ion m/z 210 as well as the fragment ions m/z 168 (M---COCH2), m/z 152 (M---NHCOCH3) and m/z 138 (M~CH2NHCOCH3) support this tentative structural assignment. The typical fragmentation pattern of the bicyclic quinolizidine ring system is represented by the fragment ions m/z 124, m/z 110, m/z 97 and m/z 83 (Sastry, 1972). The mass spectrum of dihydroammodendrine displayed a M + at m/z 210 together with a base peak at m/z 84 and a fragment ion at m/z 126, which is consistent with the fragmentation of this bipiperidyl alkaloid into two piperidyl rings. Besides the 25 identified alkaloids we detected 11 unknown compounds in the various extracts of Poecilanthe. Unfortunately, only very limited plant material was at our disposal, which was insufficient to permit the isolation and identification of the unknown compounds by standard phytochemical procedures. On the other hand, most of these unknowns represented minor constituents and their identification was not crucial for an interpretation of the alkaloidal data in terms of the relationships of the genus Poecilanthe. Table 1 shows the total yield and the distribution of alkaloids in 15 leaf extracts of 10 species of Poecilanthe. Poecilanthe amazonica, P. falcata and P. ulei accumulated the highest amounts of alkaloids, whereas P. grandiflora and P. parviflora had extremely low yield figures, so that relative yields are not given for these t w o species. Obviously the total content of alkaloid in the leaves is subject to considerable fluctuations within some species (e.g.P. hostmannii or P. itapuana; Table 1). ¢-Pyridone alkaloids were found as major constituents in leaves of P. falcata, P. itapuana, P. ovalifolia, P. subcordata, P. uleL The bicyclic quinolizidine alkaloid epilupinine was present as main compound in P. amazonica, P. falcata and P. hostmanniL Lusitanine and dihydrolusitanine, also bicyclic quinolizidine alkaloids, were found in P. arnazonica, P. effusa, P. grandiflora and P. hostmanniL Bipiperidyl alkaloids (ammodendrine, dihydroammodendrine) occur mainly in P. amazonica and

550

R. GREINWALD E T A L .

TABLE 1. DISTRIBUTION OF M A J O R ALKALOIDS IN LEAF EXTRACTS FROM 10 SPECIES OF POECILANTHE. Alkaloid distributions are given as percentages of total yield as estimated from GC results using peak area and 100 ng p1-1 sparteine as external standard. ( - - = n o t detected; + =present in trace amounts only) Distribution of alkaloids (% of total alkaloid yield) Total yield (l~gg - 1 )

1

sample 1

726

59

1

13

6

15

sample 2

415

48

42

3

2

2

P. hostmannii sample 1

28

64

+

5

7

18

sample 2

356

38

9

11

20

--

53 +

Species

2

3

4

5

6

7

8

9

10

11

X3

X5

X10

21

--

13

--

36

--

P. amazonica

P. effusa P. grandiflora

61

6

5

< 10

+

--

55

+

P. subcordata

464

+

P. falcata

556

48

3

24 114

2 2

÷

P. ovalifolia

+

88 55

12

+

10

2

11

20

17

+

+

+

9

2

28

+

4

16

5

5

6

+

2

24

1

5

+

P. itapuana

sample 1 sample 2

....

+

52

P. parviflora

sample 1 sample 2

< 10 < 10

+

+

+

P. ulei

sample 1

187

5

69

1

7

3

5

sample 2

490

3

16

10

2

41

15

1 = epilupinine;

2 = 4~-hydroxylupinine;

3 = dihydrolusitanine;

4 = dihydroammodendrine;

5 = lusitanine;

6 =N-methyl-

cytisine; 7 = cytisine; 8 = lupanine; 9 = N-formylcytisine; 10 = anagyrina; 11 = baptifoline.

P. hostmanniL The leontidine-type alkaloid camoensidine was discovered as minor constituent in both leaf samples of P. itapuana. The distribution of major alkaloids and the total yield of seed extracts from five species of Poecilanthe are shown inTable 2. The ¢-pyridone alkaloids cytisine and Nmethylcytisine were present as main constituents in seed extracts of P. parviflora, P. subcordata and R u/eL In seeds of P. parviflora we found additionally high amounts of camoensidine and of an unknown alkaloid X4, and the unknown ¢-pyridone X2 was another main alkaloid in P. subcordata. In contrast to these species no ¢-pyridone alkaloids could be detected in seeds of P. amazonica and P. effusa. Epilupinine occurred as a major compound in seeds of P. amazonica. In seeds of P. effusa we TABLE 2. DISTRIBUTION OF MAJOR ALKALOIDS IN SEED EXTRACTS FROM FIVE SPECIES OF POECILANTHE. Alkaloid distributions are given as pementages of total yield as estimated from GC results using peak area and 100 ng i~1-1 sperteine, as external standard. ( - - = Not detected; + = present in trace amounts only) Distribution of alkaloids (% of total alkaloid yield) Total yield Species P, amazonica

(p.g g--I ) 849

1

3

89

6

5

.

. .

. .

13

.

< 10

.

P, subcordata

3518 2140

+

---

20 1

57 54

1 25

5886 9660

---

----

1 1

92 91

. --

1

.

12

P. effusa P. parviflora

.

7

.

.

X2

X4

19 --

+ 11

.

.

.

-5

P. u/ei

sample 1 sample 2

.

. --

. --

1 = Epilupinine; 3 = dihydrolusitanine; 6 = N- methylcytisine; 7 = cytisina; 12 = camoensidine; 13 = teshiromine.

ALKALOIDS OF GENUS POECILANTHE

551

found only a very low yield with lupanine and the unknowns Xl0 and Xll as trace compounds. The ¢-pyridone accumulating species had clearly higher yields in the seeds than P. effusa and P. amazonica (Table 2). Discussion The presence of alkaloids in the genus Poeci/anthe was demonstrated for the first time. At least 36 different alkaloids were detected in leaves and seeds of the 10 examined species. The major constituents belong to the quinolizidine alkaloids and the bipiperidyl alkaloids. Our present data reveal a considerable variation in the alkaloids within the genus Poecilanthe. Leaves of the species P. falcata, P. itapuana, P. ovalifolia, P. parviflora, P. subcordata and R ulei accumulate the ¢-pyridone alkaloids cytisine, N-methylcytisine, N-formylcytisine, anagyrine and baptifoline as main constituents. In contrast to this, R amazonica, P. effusa, P. grandiflora and R hostmannii are characterized by the presence of bicyclic quinolizidine alkaloids (epUupinine, lusitanine, dihydrolusitanine), while ¢-pyridone alkaloids are totally absent. The bicyclic quinolizidine alkaloid epilupinine is present in both groups of species. It co-occurs with cytisine in leaves of P. falcata and P. itapuana. The total yield of alkaloids in leaf extracts varied between <0.001% dry wt. and 0.073% dry wt. (P. amazonica). Presently it is not clear if the observed variation in the alkaloid yield of the 10 species is due to environmental factors (soil, temperature, humidity) or to the growth state of the leaves (young growing leaves or old senescing leaves). Considerable intraspecific variability plays a role at least in R hostmanniL The results of the seed extracts agree well with the data on the leaf extracts contributing evidence that there are two groups of species with different alkaloid patterns within the genus Poecilanthe. Seeds of P. parviflora, P. subcordata and P. ulei revealed an ¢-pyridone profile, whereas seeds of R amazonica and P. effusa were totally devoid of ¢-pyridone alkaloids. Both groups of species are additionally separated by a striking difference in the total alkaloid content of the seeds. The species accumulating c¢-pyridone alkaloids exceeded clearly P. amazonica and P. effusa. At least from these preliminary investigations the alkaloid profile of P. effusa seemingly has some distinct features, which deserve attention in further studies. The alkaloid data confirm Lavin's findings that Poecilanthe should not be placed in the tribe Robinieae, as was suggested by Geesink (1984). So far there is no report on the presence of quinolizidine alkaloids in the Robinieae. Several important phytochemical and morphological features (no accumulation of canavanine in the seeds, anthers dimorphic, and others (Lavin, 1989); structure of root nodules (Sprent et al., 1989)) indicate a closer affinity of the genus Poecilanthe to some genera in the Dalbergieae than to any in Robinieae. The presence of alkaloids within Poecilanthe does not suggest a closer affinity to the Dalbergieae. Alkaloids are apparently not wide-spread within this tribe. There is one report on the occurrence of quinolizidine alkaloids in the species Dalbergia monetaria L. fil. (Kinghorn et aL, 1982). Attention should rather be paid to Lavin's remark, that the dehiscent legumes possessed by some species of Poecilanthe with a spongiose inner epidermis and dimorphic anthers are reminiscent of Brongniartieae. Dimorphic anthers are also present in the Templetonia group of the former tribe Bossiaeeae (Crisp and Weston, 1987). In Harpalyce (Kinghorn et aL, 1982) and Lamprolobium (Greinwald et al., 1993) as well as in Templetonia biloba (Greinwald et aL, submitted), all members of the Brongniartieae, the co-occurence of epilupinine and cytisine has been reported. The co-occurrence of these alkaloids was also found for Poecilanthe. Concerning the alkaloid profile, a connection of Poecilanthe can also be shown to the tribe Sophoreae. Especially the genera Dicraeopetalum and Sakoanala (van Wyk et aL, 1993) and Maackia (Ohmiya et al., 1991) exhibited distinct similarities with Poecilanthe.

552

R. GREINWALD ETAL.

The alkaloidal data for Poecilanthe conflict with the present taxonomic position of this genus and should initiate a thorough re-examination of its generic relationships. More detailed studies are necessary, to get further information on the alkaloid distribution in different plant parts (seeds, pods, flowers) and to elucidate the structure of the unidentified alkaloids. Acknowledgements---We thank Prof. W. Keller (School of Pharmacy, Northeast Louisiana University, Monroe) for providing us with an authentic sample of lusitanine, Prof. B.-E. van Wyk (Dept. of Botany, Rand Afrikaans University, Johannesburg) for reference samples of anagyrine and thermopsine and the Prof. P. A. Grieco (Department of Chemistry, Bloomington, Indiana, U.S.A.) for reference samples of lupinine and epilupinine.

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