Phymhemisrry, Vol. 26, No. i 1,pp.3021-3023, 1987. 0031~%22/87 $3.00+ 0.

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Printed in Great Britain. Pertpawn Journ& Ltd.

NIMBIDIOL, A MODIFIED DITERPENOID OF THE ROOT-BARK OF

AZADIRACHTA INDICA

P. L. MAJUMDER, D. C. MAITI, W. KRAUS* and M. YOKEL*

Dqwtment of Chemistry, Uaiversity Colkgc of S&ace, Calcutta Mooo9, Iwiia; *Institut fiir CheaGe 130, Lehrstuhl fiir Orgaakhe
Chcmie aa der Universitit Hohe&eim, Stuttgart, F.R.G.

(Receilted27 February 1987)

Key Word Iadex--ilzodiruchtn indica; Meliaceac; Indian ‘ncem’; modified diterpem; nimbidid.

Abstract-Nimbidiol, a modified diterpeooid, isolated from the root-bark of Azadirachta indicn (Indian ‘neem’), was
characterized by spectroscopic method.

I~ODU~ON hydroxyl functions in in~~ol~u~ hydrogen bonding.

The spectrum of aimbidiol exhibited signals for only two
Azudiraclttu in&a A. Juss, popularly known as &em’ in
aromatic protons at 66.85 and 7.39 (each lH, s) indicating
India, has been the subject of extensive chemical investi- the tetrasubstituted nature of its benxenoid ring. The
gation by several groups of workers [l-14] because of its
chemical shift of the proton cqesponding to the signal at
varied biological activities [S-20]. Of particular import-
67.39 was consistent with its being flanked by a carbonyl
ante is the isolation of the antifecdant azadirachtin [8]
and a hydroxyl group, and the multiplicity of the two
and several of its structural analogues [ 141 l] from the
aromatic proton signals suggested that the two aromatic
leaves and fruits of this plant. There are, however, few
protons in nimbidial were pore to each other and were
reports of the chemical investigation of its root. This has
each ortho to a hydroxyl group. This was borne out by the
prompted us to undertake a systematic chemical investi-
fact that the signals at 66.85 and 7.39 of nimbidiol were
gation on the root of A. in&a, the root-bark of which
shifted downfield to 67.19 and 7.80 respectively in the
has yielded a new modified diterpenoid (la), which we
‘HNMR spectrum of nimbidiol diacetate. In the
have named nimbidiol.
‘H NMR spectrum of the monoacetate only the signal at
67.39 was -&i&d downfield to a7.67, the &nal for the
RESULTS AND DISCU~ION other aromatic proton remaining essentially unchanged
Nimbidiol (la), C1,HZZ03 (EM’] = m/z 274),showed
UV absorptions [ASH 210, 238, 282 and 323 nm (log
84.19, 4.12, 3.91 and 3.78)] strikingly similar to those of
nimbi01 (le) [3] and sujiol (If) [S] indicating the presence
of a phydroxybenxoyl chromophore. Its phenolic nature
was indicated by its characteristic colour reactions, by the
alkali-induced bathchromic shift of its UV maxima
[AEHa.l NtioH 211, 256 and 353 nm (loge4.40, 3.99
and 4.23)] and by the presence of bands at 3500,342O and
3230 cm- l in its IR spectrum. The presence of these three
bands, particularly the last one, suggested that the hydro-
xyl groups were pa&lly hydro~-Andy. The IR
spectrum of nimbi~ol also revealed the presence of an R’ I R3 R”
aromatic keto-carbonyl function (v_ 16SOcm-‘). The
ia OH OH H 0
absorption of this carbonyl group at a relatively high
wavelength may be attributed to the presence of a lb OH OAC H 0
hydroxyl function pare to the carbonyl group. The Ic OAC OAc H 0
presence of two phenolic hydroxyl groups in nimbidiol ld or& OMe H 0
was confirmed by the formation of a monoacetate, le OH Me H 0
C,,H,,O, ([Ml’ -m/z 316), a diacetate, Cz1Hz60s If OH -CH(Me)~ H 0
(CM]” = m/z 358), and a dimethyl ether derivative, 0
II! H H -CH(Me)2
&Hz60J (CM]’ = m/z 302).
lb OMe H H HZ
The ‘HNMR spectrum of nimbidiol showed a two-
proton broad signal (disappeared on deuterium exchange) Ii O& ---cH(Me)a H HZ
at 69.22 corroborating the presence of two phenolic 11 -CfiWeh OMS H Hl
hydroxyl groupa in the compound. The downfield pos- IL OAC ---CH(Me)t H H2

ition of this signal also indicated the involvement of these II H OAC -CH(I& H¶
3022 P. L. MNUMDER et al.

(66.89). This suggested that in nimbi~ol monoacetate of C-9 by w 5-6 ppm compared to the ~~~~n~ng
only the hydroxyl group ortho to the aromatic proton carbon atoms of 1 h-l I having a methylene group at C-7.
corresponding to the signal at 67.39 of nimbidiol had The appearance of the methoxyl carbon resonance of Id at
undergone acetylation. The ‘HNMR spectrum of nim- S, 55.90 indicated that each methoxyl group had at least
bidiol also displayed three three-proton singlets at b0.93, one ortho hydrogen atom, and the upfield shifts of C-8, C-
1.00 and 1.21 for three methyl groups attached to sp3 9, C-11 and C-14 of ld compared to those of the
carbon atoms, and a two-proton multiplet centred at corresponding carbon atoms of 1~ conlirmed the place-
62.54 for the protons of a keto-methylene group of the ment of the two hydroxyl groups of nimbidiol at C-12 and
type>CHX!H&O-C~. The above spectral features were C-13. The difference in the G,values of C-8 and C-l 1 of the
also discernible in the ‘H NMR spectra of ail the deriva- mono and diacxtyl derivatives of nimbidio1 was consistent
tives of nimbidiol. Based on the foregoing spectral data with the pbment of the hydroxyl group in the mono
nimbidiol was assigned the structure la, and its mono and acetyl derivative at C-12.
diacetyl and dimethyl ether derivatives structures lb, lc Nimbidiol is thus a unique modified diterpenoid of the
and Id respectively. abiatane skeleton lacking an isopropyl group at C-13.
More compelling evidence in support of the above Biogeneti~lly it may be assumed to be formed from an
structure of nimbidiol was provided by the r3CNMR abiatane skeleton by an unknown oxidative process.
spectral analysis of its more soluble monoacetyl (lb),
diacetyl (lc) and dimethyl ether (Id) derivatives. The
degree of protonation of the carbon atoms in each
compound was determined by DEPT experiments and the
assignments of the carbon chemical shifts (Table 1) were Mps: uacorr; CC:silica gel (60-120 mesh); TLC: .siliorgel G;
made by comparison with the 6, values of sujiol (lf) UV: 95% sldehyde-free EtOH, IR: KBr; ‘H NMR: 80 MHz,
[21,23] and those of its structural analogues lg-11 CD&, TMS as un. star&r&, 13CNMR: 62.5 MHr, CDQ,
[21-231 taking into ~n~deration of the additive para- TMS; MS: direct inlet, 7OeV. AI1 the anaIytical samples were
meters of the functional groups. Thus the S, values of C-l, routinely dried over Pros for 24 hr in racuo and were tested for
C-7, C-10, C-18, C-19 and C-20 of lb, lc and Id were purity by TLC and mass spectrometry. Na#O* was used for
almost identical with those of the corresponding carbon drying organic solvents and the petrol used had bp 6@-80°.
atoms of sujiol (If) [21,23] and (lg) [21,23] confirming Isokztioo o~n~~io~ (la). Air-dried powdered root-bark (2 kg)
the structural identity of the rings A and B part of their was extracted with CHC13in a Soxhkt app&atus for 72 hr. The
molecules, The presence of a conjugated keto-carbonyl CHC& extract was cotmd (15OmI) and subjected to CC. The
function in all the compounds was indicated by the signals petroI-EtOAc (5: 1) ehrate afforded a gummy residue oontaitting
at S, 198.40,197.42 and 198.12 and its location at C-7 was mainly la. Repeated chromatography of the above material gave
corroborated by the downfield shift of C-6 of all the Pure la (0.2 g), crysM from a mixture of petrol, EtOAc and
compounds by N 17 ppm and a concomitant upfield shift C*H+ mp 226”, [aIn+ 3.4” (CHCl,), (Found: C., 74.39; H, 8.10;
Ci7Hr203 requires: C, 74.45; H, 8.03%). MS: m/z 274 [Ml+
(loo%), 259 (97), 245 (5), 217 (23), 203 (13), 191(70), 189 (71), 177
Table 1. %NMR spectral data of monoacetyl nimbidial (lb), (44), 163 (19), 151(6X 137 (6), 115 (8), 83 (6), 77 (a), 69 (27), 55 (11).
diacetyl nimbidiol (1~)and dimethyl ether of nimbidiof (ld) and 41 (20).
Nimbidiol nwnoacetate (lb), nimbidiol diucetate (lc) and nint-
(6 values) bidiol dimethyl ether (ld). Nimbidiol (0.15 g) was acetylated with
Ac10 and CsH,N to give a mixture of lb and lc. On CC of the
lb lc Id mixture the petrol-EtOAc (30: 1) eluate gave lc (0.1 g), crystal-
lized from petroi_EtOAc, mp 160°, [aJD + 2.2” (CHCI,), (Found:
1 37x0* 37.84 38.04 C, 70.41; H, 7.20; C2,Hr605 requires: C, 70.39; H, 7.26%).
2 18.75 18.68 18.84 UVI ,nm: 210, 253, 290 (log& 4.32, 4.09 and 3.46)
3 41.20 41.15 41.28 IRv,cm- ‘: 1775 and 1260 (OAc), 1680 (conjugated GO),
4 33.20 33.24 33.21 1600,8l!O, 860 and 820 (phenyl nucleus); ‘H NMR: S7.80 (lH, s,
5 49.29 49.13 49.90 H-14),7.19 (lH, s, H-11), 2.60 (2H, m, H,-6), 2.29 and 2.28 (each
6 35.70 35.86 35.85 3H, s, 2 x OAc), 1.25,0.99 and 0.94 (each 3H, s, 3 x C-Mek MS:
1 198.40 197.42 198.12 m/z 358 [M]* (18x), 316 (12), 274 (lOa), 259 (41), 217 (6), 191
8 124.45 129.50 124.20 (15),189(18),177(14~163(6~91(5~84(8~69(15),55(13~49(35)
9 152.85 154.9 147.34 and 43 (62).
10 38.03 38.14 38.04 The petrol-EtOAc (20: 1) eluate gave lb (0.045 g), crystallixed
11 112.40 119.16 105.64 from petrol-EtOAc, mp 220”. (Found: C, 71.99, H, 7.62.
12 156.50 146.42 153.84 C1sHz404 requires: C, 72.15; H, 7.59%). UV L, nm: 210,254
13 136.90 140.4 151.27 and 295 (log E 4.27,4.03 and 3.30); IR v, cm-‘: 3400 {OH), 1770
14 121.90 122.30 108.67 and 1265 (OAc), 1655 (conjugated c--O), 1590,885,870 and 820
18 32.45 32.44 32.40 (phenylnucleus); ‘H NMR: 67.67 (lH,s,H-14),6.89 (lH,s, H-11),
19 21.28 21.23 21.25 2.69 (2H,m, Hx-6), 2.31(3H,s, OAc), 1.20,0.97and0.92 (each 3H,
20 23.16 23.31 23.16 s, 3 x C-Me); MS: m/z 316 [Ml+ (lS”/,x 274 (lOO), 259(63), 217
-0COMe 169.35, 20.80 168.09, 167.63 - (13),203(P), 191(38), 189 (44), 177 (lo), 151(6), 137 (6), 113 (9x91
20.63, 20.49 (9 83 (lo), 77 (9). 69 (36), 55 (31), 49 (lo), 43 (64) and 41 (37).
- OMe - - 56.90 A soln of la (0.05 g) in MeOH (25 ml) was treated with an
excess of an ethereal soln. of CHZN2 in the cold. The reaction
* Values are in ppm downfield from TMS: B(r~s) = Btc~t,,) mixture was kept overnight and then the solvent was removed
+ 76.9 ppm. under red. prea. On CC of the residue, the petrol-EtOAc (30: 1)
 Nimbidiol, a modified diterpenoid of the root-bark of Azadirachta indica 3023

eluate 8ave Id (0.045 g), crystallized from petrol-EtOAc, mp 112” 9. Banerji, R., Misra, G. and N&am, S. K. (1977) Fitoterpia 48,
(Found: C, 75.61; H, 8.55. C19H2603 requires: C, 75.50, H, 8.61%). 166.
‘HNMR: 67.41 (lH, s, H-14), 6.73 (lH, s, H-11), 3.87 and 3.84 10. Kraus, W., Cramer, R. and Sawitzki, G. (1981) Phyto-
(each 3H, s, 2 x OMe), 2.58 (2H, m, H,-6), 1.20,0.% and 0.88 chemistry 20, 117.
(each 3H, s, 3 x C-Me) MS: m/z 302 [M]’ (17 %), 287 (69), 259 11. Bruhn, A., Bokel, M. and Kraus, W. (1984) Tetrahedron
(6), 245 (30), 231 (19), 219 (100x 217 (9i), 205 (48), 203 (16), 191 Letters 3691.
(33), 189 (15), 165 (14),163(6),145(11X 130(12), 128 (14), 115 (24), 12. Gar& H. S. and Bhakuni, D. S. (1985) Phytochemistry 24,866.
93(31),82(18),71(13),69(31),67 (47),55 (31),49(18)and43 (46). 13, Podder, G. and Mahato, S. B. (1985) Heterocycles 23,232l.
14. Devkumar, C. and Mukherjee, S. K. (1985) Indian J. Chem.
Acknowledgements-We thank Dr J. M. Wilson, Univerisity of 24B, 1105.
Manchester, for the mass spectra. The work was supported by 15. Ibrahim, A. and Dorak, H. (1984) Envier. Entamol. 13,803.
CSIR, New Delhi, India. 16. Kubo, I., Matsumoto, A., Matsumoto, T. and Klocke, J. A.
(1986) Tetrahedron 42,489.
17. Rajanopo, W., Suwanno, S., Somjaru, R., Glinsukon, T. and
Thebtaranont, Y. (1985) J. Sci. Sot. Thailand 11, 177.
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