Synthesis and biological evaluation of
novel bile acid-nucleoside conjugates
published results and work in progress
Maria Luisa NAVACCHIA
CNR ISOF
synthesis and characterization
of modified nucleosides
CNR-ISOF
Massimo Capobianco
Maria Luisa Navacchia
•synthesis and characterization of
modified bile acids
• click-chemistry
UNIFE-Dipartimento di Scienze Chimiche e Farmaceutiche
Daniela Perrone
Elena Marchesi
Lara Mari
biological evaluation
UNIFE-Dipartimento di Scienze Chimiche Mediche
Riccardo Gavioli
Fabio Sforza
Synthesis of: 3a-azido-bile acids and
8-alkynylic-2’-deoxyadenosines
Conjugate synthesis via click-chemistry
i: CuSO4 . 5 H2O, sodium ascorbate, THF–tBuOH–H2O (1.5 : 1 :1), 25 °C, 18 h, 70%; or
microwave 80 °C, 30 min, 73%.
Synthesis of 8-alkynylated-2’-deoxyadenosines
NH2
N
HO
O
N
N
Br
N
NH2
N
S
3
N
N
N
HO
H2O, TEA
+
3
SH
O
100 °C, 2 h
OH
OH
This reaction can be smoothly performed in water and leads to a simple purification of the thioalkylnylated
nucleoside that only requires the extraction of the compound with warm EtOAc from the aqueous crude mixture.
M. L. Capobianco and M. L. Navacchia PCT Int. Appl. WO 2012164484 A1 2012
NH2
N
N
N
O
n
+
OH
N
n
N
Br
HO
NH2
n=3
Pd(PPh3)2Cl2
CuI
DMF
50 °C
N
HO
N
N
O
OH
This reaction can be smoothly performed and leads to a simple purification of the alkylnylated
nucleoside that requires a filtration on florisil.
Biological evaluation
in vitro cytotoxicity toward human fibroblast cells
and anti-proliferative activity against four human
cancer cell lines: Leukemic T Jurkat and K562, colon
carcinoma HCT116 and ovarian cancer A2780
Cytotoxic activity of bile acid-based conjugates and
alkynyl deoxyadenosines dA-A, HdA-A, and SdA-A on
human cancer cell lines and human fibroblast cellsa
Anti-proliferative activity of conjugated
compounds against the K562 cell line
Percentage of apoptotic K562 cells determined after
24 h treatment with HdA-CDC, SdA-CDC and HM-CDC
(50–10 mM) by annexin V staining.
Conclusion
OMe
• Best activity was shown by CDCbased derivatives and could be
correlated to the lipophilicity and to
the 7a-OH group orientation;
O
OH
X
• furthermore, except dA-UDC and HM-bile acid series, all new
conjugates did not show any significant cytotoxicity towards
the human fibroblast cells whereas some of them strongly and
selectively inhibited cell proliferation and induced apoptosis in
leukemic K562 cells;
• therefore, these derivatives constitute a starting lot of
candidate drugs.
…however many questions are still open
Do other nucleobases work?
Does the ribo form work?
How much important is the triazole ring?
NH2
N
n
O
n
N
n=3
N
HO
n=3
N
N
N
NH
NH2
HO
O
OH
OH
H-A-A
O
n=3
NH
HO
O
OH
N
n
N
O
H (OH)
O
OH H (OH)
H-dU-A, H-U-A
H-dG-A, H-G-A
click
H
N
OMe
O
O
OH
OH
N3
N3
N3
N3-CDC 7aOH
N3-UDC7-OH
OH
OH
Nor-23-N3-CDC 7aOH
Nor-23-N3-UDC7-OH
N3-TUDCA
SO3
2
NH2
N
N
Br
N
HO
O
NH2
HO
N
SH
H2O, TEA
+
OH
OH
OH
100 °C, 2h
N
N
S
OH
N
HO
O
OH
Nor-23-SH-CDC 7aOH
Nor-23-SH-UDC7-OH
A new lot of 27 bioconjugates
is waiting for the biological assay
N
References
Adenosine Or Deoxyadenosine Derivatives Modified At Position 8 And A Method Of
Synthesis Thereof ;
Massimo L. Capobianco and Maria Luisa Navacchia PCT Int. Appl. WO 2012164484 A1 2012.
Labeling Deoxyadenosine for the Preparation of Functional Conjugated Oligonucleotides;
Massimo L. Capobianco,* Elena Marchesi, Daniela Perrone and Maria Luisa Navacchia
Bioconj., 2013, 24, 1398-1407.
Synthesis and in vitro cytotoxicity of deoxyadenosine-bile acid conjugates linked with 1,2,3triazole;
Daniela Perrone,* Olga Bortolini, Marco Fogagnolo, Elena Marchersi, Lara Mari, Chiara
Massarenti, Maria Luisa Navacchia,* Fabio Sforza, Katia Varani and Massimo Luigi
Capobianco New J. Chem., 2013, 37, 3557-3557.