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Structures and evaluation of biologically active constituents of "Cussonia Zimmermannii" harms

Senn, Martin W.. Structures and evaluation of biologically active constituents of "Cussonia Zimmermannii" harms. 2006, Doctoral Thesis, University of Basel, Faculty of Science.

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Official URL: http://edoc.unibas.ch/diss/DissB_7526

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Abstract

For the present thesis 22 samples of seven African medicinal plants, the extracts of
which had previously proved to possess an in vitro activity against T. b. rhodesiense
and/or P. falciparum and 13 samples of six randomly selected plant species based on
availability were collected in Tanzania.
Of these 35 samples, 140 crude extracts were produced with four solvents of
different polarity, and the extracts were tested for in vitro antitrypanosomal and
antiplasmodial activity and for cytotoxicity. In addition the extracts were tested for the
affinity to the GABAA receptor performed by radioligand binding experiments.
Based on the considerable antitrypanosomal and antiplasmodial activity and the
affinity to the GABAA receptor, two extracts were selected for bioassay-guided
fractionation : the petroleum ether extract of the stembark of Commiphora
fulvotomentosa Engl. because of its promising IC50 value of 2.1 mg/ml against T. b.
rhodesiense and the high selectivity index of 21.4, and the petroleum ether extract of
the rootbark of Cussonia zimmermannii Harms because of its IC50 value of 4.8 mg/ml
against T. b. rhodesiense, its IC50 value of 3.3 mg/ml against P. falciparum, and the
potent modulatory effect at the GABAA receptor of 151 %.
After three fractionation steps the fractions of the petroleum ether extract of the
stembark of Commiphora fulvotomentosa Engl. did not show any inhibitory activity
against T. b. rhodesiense and were therefore not investigated further.
Bioassay-guided fractionation of the petroleum ether extract of the rootbark of
Cussonia zimmermannii Harms lead to the isolation of four polyacetylenes. By the
application of MS, HR-MS, UV/VIS and IR methods and NMR experiments (1H-NMR,
13C-NMR, DEPT135, 1H-1H COSY, HMQC and HMBC) the structures of the four
novel diynes were established : 8-Hydroxyheptadeca-4,6-diyn-3-yl acetate (MS-1
(25)), 8-Hydroxyheptadeca-1-ene-4,6-diyn-3-yl acetate (MS-2 (26)), 16-Acetoxy-11-
hydroxyoctadeca-17-ene-12,14-diynyl acetate (MS-4 (27)), and 11,16-Diacetoxyoctadeca-
17-ene-12,14-diynyl acetate (MS-5 (28)).
Additionally, stigmasterol (42) was isolated and identified by comparison of its
spectroscopic data with those of an authentic sample.
For the determination of the absolute configuration of MS-4 (27) at C(11) the Mosher
method was used. The negative �d values for the neighbouring protons suggested
the S-configuration. But this result could not be confirmed by the 13C-NMR data since
both positive and negative �d values were obtained for the neighbouring carbons.
Due to the fact, that the results for the protons and the carbons were not consistent
the S-configuration for C(11) of MS-4 (27) could not be assigned with certainty.
The isolated compounds MS-1 (25), MS-2 (26), and MS-4 (27) were tested for in vitro
inhibitory activity against additional parasites like T. cruzi and L. donovani (axenic
and in infected macrophages). 42 was not tested because it was a well known
phytosterol which is ubiquitous in plants. Neither was MS-5 (28) tested, since the
isolated amount was not sufficient.
It was found that MS-1 (25) showed in all antiparasitic in vitro assays no higher
inhibitory activity than the crude extract. MS-2 (26) showed promising activities in the
T. cruzi and L. donovani (axenic and in infected macrophages) assay with IC50 values
of 0.2, 0.039, and 0.098 mg/ml, respectively. The respective IC50 values of the
standard drugs were 0.62, 0.18, and 0.29. The cytotoxicity was relatively high;
therefore the selectivity with SI values of 18.0 and 36.7, respectively (T. cruzi, L.
donovani in inf. mac.), was in a moderate range. MS-4 (27) showed also interesting
activities in the T. cruzi and L. donovani (axenic) assays with IC50 values of 0.15 and
0.054 mg/ml. The SI value of 145.3 (T. cruzi) was high. Compared with the standard
drugs the activities of MS-2 (26) and MS-4 (27) against T. b. rhodesiense and P.
falciparum were in a moderate range.
MS-1 (25), MS-2 (26), and MS-4 (27) were also tested in the GABAA receptor binding
assay. Here MS-4 (27) showed the highest relative specific binding of 158 % at a
concentration of 20 mg/ml, followed by MS-2 (26) with 152 % and MS-1 (25) with
138 %.
In order to investigate whether the modulatory effect at the GABAA receptor leads to
a chanel opening, the isolated compounds were investigated electrophysiologically. It
was found that MS-1 (25), MS-2 (26), and MS-4 (27) act as potent positive allosteric
modulators at GABAA receptors with a half maximal stimulation at a concentration of
0.6-3.5 mM and a maximal stimulation of 110-450 %. The compounds also showed
unique subunit selectivity profiles, the stimulation was independend from the
presence of the g subunit and resistant to the benzodiazepine antagonist Ro15-1788.
These in vitro data suggested that MS-1 (25), MS-2 (26), and MS-4 (27) may be used
to treat diseases of the central nervous system, specifically they may be used to treat
states of anxiety, as sedatives/hypnotics, as muscle relaxants and as anticonvulsives
(e.g. in epilepsy), and in the treatment of drug addiction. Therefore it was decided to
patent the compounds for these indications [174].
All data are on the in vitro level. For a pharmaceutical application in vivo data are
needed. Therefore MS-1 (25), MS-2 (26), and MS-4 (27) need to be synthesized in
order to get more material for subsequent in vivo experiments.
Advisors:Séquin, Urs Marcel
Committee Members:Brun, Reto and Hamburger, Matthias Otto
Faculties and Departments:05 Faculty of Science > Departement Chemie > Former Organization Units Chemistry > Physikalische Chemie (Maier)
UniBasel Contributors:Brun, Reto
Item Type:Thesis
Thesis Subtype:Doctoral Thesis
Thesis no:7526
Thesis status:Complete
Number of Pages:148
Language:English
Identification Number:
edoc DOI:
Last Modified:22 Jan 2018 15:50
Deposited On:13 Feb 2009 15:36

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