Thesis (MSc)--Stellenbosch University, 2002. / ENGLISH ABSTRACT: The aim of this thesis was to define the interactions of the androgen receptor
(AR) with an analog of a non-steroidal plant compound, Compound A (CpdA), as
well as two synthetic progestins, medroxyprogesterone acetate (MPA) and
norethindrone acetate (NET-A). The data presented indicates that CpdA has antiandrogenic
properties, as it represses androgen-induced activation of both specific and
non-specific androgen-responsive reporter constructs. It was found that CpdA exerts
these effects by a mechanism other than competition with androgen for binding to the
ligand-binding domain (LBD) of the receptor. On the other hand, it is demonstrated
that both MPA and NET-A compete with androgen for binding to the AR and induce
partial agonist activity via the receptor. Using mammalian two-hybrid assays it was
revealed that CpdA, similar to anti-androgenic compounds that are able to compete
with androgens for binding to the receptor, represses the androgen-induced interaction
between the NH2- and COOH-terminals of the AR (N/C-interaction) without
competing for binding to the LBD. Furthermore, it was shown that CpdA slightly
represses the androgen-dependent recruitment of steroid receptor co-activator 1
(SRC1) to the activation function (AF2) domain of the AR. When the effects of MPA
and NET-A on the N/C-interaction were studied, intriguing results were obtained.
NET-A, as expected, induced this AR agonist-induced interaction. MPA, however,
repressed this AR agonist-induced interaction, an effect previously associated with
anti-androgenic activity, despite displaying partial agonist activity in transctivation
experiments. On the other hand, both MPA and NET-A induced the interaction
between SRC1 and the AF2 domain. In additional experiments with CpdA, it was
found that CpdA did not affect the recruitment of SRC1 to the AF1 domain of the
receptor; neither did it influence the constitutive activity of the NH2-terminal domain.
The anti-androgenic activities of CpdA were confirmed by the toxic effect that this
compound had on the androgen-dependent lymph node carcinoma of the prostate
(LNCaP) cell-line as well as its ability to repress the androgen-induced expression of
the prostate specific antigen (PSA) protein. Taken together, the results presented in
this thesis, in combination with the knowledge available on AR function, contribute to
an improved understanding of AR function. Furthermore, the importance of defining
the precise mechanism by which individual compounds exert their effects is
highlighted. In this regard it is demonstrated that two compounds (MPA and NET-A)
that display partial agonist activity, can exert their effects via different mechanisms at
the molecular level. Detecting such differences in the molecular mechanisms of action
could facilitate the improved design of progestins as well as aid clinicians and their
patients in selecting the best method of contraception. Lastly, the insights gained into
the mechanisms of the anti-androgenic action of CpdA could be useful in therapeutic
drug design for diseases, such as prostate cancer, that have an androgen-dependent
etiology. / AFRIKAANSE OPSOMMING: Die doel van hierdie tesis was om die interaksies van die androgeen reseptor
(AR) met ‘n analoog van ‘n nie-steroiediese plant verbinding, Verbinding A (VbgA),
sowel as met twee sintetiese progestogene, medroksiprogesteroon asetaat (MPA) en
noretiendroon asetaat (NET-A), te definieer. Die data verskaf dui daarop dat VbgA
anti-androgeniese eienskappe besit deurdat dit androgeen-gei'nduseerde aktivering van
beide spesifieke- en nie-spesifieke androgeen-responsiewe rapporteerderkonstrukte
onderdruk. VbgA veroorsaak hierdie effekte deur ‘n meganisme wat nie kompetisie
met androgeen vir binding aan die ligand-bindingsdomein (LBD) van die reseptor
behels nie. In teenstelling hiermee word getoon dat beide MPA en NET-A kompeteer
met androgeen vir binding aan die AR en gedeeltelike agonistiese aktiwiteit induseer
via hierdie reseptor. Deur gebruik to maak van ‘n soogdier twee-hibried essai word
getoon dat VbgA, soos ander anti-androgeniese verbindings wat kompeteer met
androgeen vir binding aan die reseptor, die androgeen-gei'nduseerde interaksies tussen
die NH2- en COOH-terminale van die AR (N/C-interaksie) onderdruk, sonder om te
kompeteer vir binding aan die LBD. Daarby is dit bewys dat VbgA die androgeenafhanklike
werwing van steroied reseptor ko-aktiveerde 1 (SRC1) na die aktiverings
funksie (AF2) domein van die AR gedeeltelik onderdruk. Die studie van die effekte
van MPA en NET-A op die N/C-interaksie het interessante resultate opgelewer. NETA,
soos verwag, het hierdie AR agonis-gei'nduseerde interaksie geinduseer. MPA, aan
die ander kant, het hierdie AR agonis-gei'nduseerde interaksie onderdruk, ‘n effek wat
tevore met anti-androgeniese aktiwiteit geassosieer is, al het die transaktiveringseksperimente
daarop gedui dat MPA ‘n AR agonis is. Aan die ander kant, het beide
MPA en NET-A die interaksie tussen SRC1 en die AF2 domein geinduseer. In
addisionele eksperimente met VbgA is gevind dat VbgA geen effek het op die
werwing van SRC1 na die AF1 domein van die reseptor nie en ook geen invloed het
op die konstitutiewe aktiwiteit van die NHh-terminaal domein nie. VbgA se antiandrogeniese
eienskappe is bevestig deur die toksiese effekte op die androgeenafhanklike
limfknoop karsinoom van die prostaat (LNCaP) sellyn sowel as deur sy
vermoe om die androgen-gei'nduseerde uitdrukking van die prostaat spesifieke
antigeen (PSA) protei'en te onderdruk. Die resultate aangebied in hierdie tesis, in
kombinasie met die beskikbare kennis oor AR funksie, dra by tot ‘n verbeterde kennis
van AR funksionering. Verder word die belang van die definiering van die
meganisme waardeur individuele verbindings hulle effekte veroorsaak, getoon. In
hierdie verband is getoon dat twee verbindings (MPA en NET-A), wat gedeeltelike
agonistiese aktiwiteit besit, hulle effekte via verskillende meganismes op die
molekulere vlak veroorsaak. Deur hierdie verskille in die molekulere meganismes van
aksie uit te wys, kan beter progestogene ontwikkel word, en verder sal dit vir dokters
en hul pasiente help om die beste voorbehoedmiddel te kies. Laastens, die insig wat
verkry is ten opsigte van die meganismes van anti-androgeniese aktiwiteit van VbgA
mag nuttig wees in die ontwerp van terapeutiese middels vir die behandeling van
siektetoestande met androgeen-afhanklikke etiologie (bv. prostaatkanker).
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:sun/oai:scholar.sun.ac.za:10019.1/52683 |
| Date | 12 1900 |
| Creators | Tanner, T. M. |
| Contributors | Hapgood, J. P., Louw, A, Stellenbosch University. Faculty of Science. Dept. of Biochemistry. |
| Publisher | Stellenbosch : Stellenbosch University |
| Source Sets | South African National ETD Portal |
| Language | en_ZA |
| Detected Language | English |
| Type | Thesis |
| Format | 127 p. : ill. |
| Rights | Stellenbosch University |
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