activity,
µM
discovered five new 5-halogeno-heteroarylpyrimidines analogs
that displayed anti-HBV activity in the low micromolar range.
Despite some toxicity observed in certain cell lines, further
modifications are currently being investigated and will be subject
of future publications.
EC50
>10
>10
>10
HepG2
>100
>100
14
PBM
65
CEM
15
Vero
43
21a
21b
21c
44
23
>100
>100
Acknowledgments
20
3
21d
21e
21f
5.3 ±
3.2
This work was supported in part by NIH grant 5P30-AI-50409
(CFAR), and the National Center for Advancing Translational
Sciences of the National Institutes of Health UL1TR000454. Dr.
Schinazi is the Chairman and a major shareholder of Cocrystal
Pharma, Inc. Emory received no funding from Cocrystal Pharma,
Inc. to perform this work and vice versa.
>100
>100
>100
>100
>100
39
56
>10
>100
>10
>100
67
47
≥ 100
HAP-12
GLS4
-
-
-
0.45
0.3
>100
>100
>10
>100
28
>100
69
>100
18
3TC
0.04 ±
0.03
>100
>100
>100
References and notes
ND: not determined
In our last series of compounds (Table 3), various
modifications at the 6-position were explored. Unfortunately,
replacement of the methyl or ethyl ester of HAP-12 or GLS-4
with a thioester (26c), a trifluoroethyl ester (26e), a N-(N,N-
dimethylsulfamoyl)carboxamide (26f) or an acetamide (28a-b)
was detrimental to the anti-HBV potency of these compounds
and none of them showed activity at concentration up to 10 µM.
Interestingly, introduction of an iodine or a bromine atom at this
position lead to the discovery of compounds 26a-b and 26g-i
which displayed EC50 values between 4.0 and 5.7 µM. Chlorine,
however, was not a suitable modification and 26c was found to
be inactive at 10 µM.
1 World Health Organization website. Hepatitis B.
2016,
Accessed
10/31/2016.
2
Hepatitis B foundation. Drug watch, compound in development for chronic
hepatitis B.
Accessed 10/31/2016
2016,
3 Nassal, M. Future Vir. 2009, 4, 55.
4 Qiu, L. P.; Chen, L.; Chen, K. P. Fundam. Clin. Pharmacol. 2014, 28, 364.
5
Stoltefuss, J.; Goldmann, S.; Stolting, J.; Kramer, T.; Schlemmer, K.-H.;
Niewohner, U.; Graef, E.; Lottmann, S.; Deres, K.; Weber, O. PCT patent
WO99/54326, 1999.
6
Bourne, C.; Lee, S.; Venkataiah, B.; Lee, A.; Korba, B.; Finn, M. G.;
Zlotnick, A. J. Virol. 2008, 82, 10262.
7
Wu, G.; Liu, B.; Zhang, Y.; Li, J.; Arzumanyan, A.; Clayton, M. M.;
Table 3. HBV inhibition and cytotoxicity of series IV: compounds 26a-i and
28a-b.
Schinazi, R. F.; Wang, Z.; Goldmann, S.; Ren, Q.; Zhang, F.; Feitelson, M.
A. Antimicrob. Agents Chemother. 2013, 57, 5344.
8
Zhu, X.; Zhao, G.; Zhou, X.; Xu, X.; Xia, G.; Zheng, Z.; Wang, L.; Yang,
X.; Li, S. Bioorg. Med. Chem. Lett. 2010, 20, 299.
9 Han, J.; Pan, X.; Gao, Y.; Weil, L. Virus Res. 2010, 150, 129.
10 Bourne, C. R.; Finn, M. G.; Zlotnick, A. J. Virol. 2006, 80, 11055.
11
Bassit L.; Ozturk T.; Tao S.; Boucle S. R.; Ehteshami M.; Zlotnik A.;
Feitelson M. A.; Schinazi R. F. Abstract of paper, 48th Annual meeting of the
European Association for the Study of the Liver, Amsterdam, Netherland,
2013, Abstract 736.
Anti-HBV
activity,
µM
Cytotoxicity, CC50 (µM)
Cmpd
R1
R2
R3
R4
12
Billioud, G.; Pichoud, C.; Puerstinger, G.; Neyts, J.; Zoulim, F. Antivir.
EC50
4.1
HepG2
>100
PBM
80
CEM
74
Vero
29
Res. 2011, 92, 271.
13
Lam, A.; Ren, S.; Vogel, R.; Espiritu, C.; Kelly, M.; Lau, V.; Hartman, G.
Cl
Cl
Cl
Cl
H
H
H
H
I
26a
26b
26c
26d
D.; Flores, L.; Klumpp, K. AASLD Liver Meeting 2015. San Francisco,
November 13–17, 2015. Abstract 33.
14
Br
Cl
5.2 ± 1.0
>10
79
98
>100
23
44
13
16
13
40
Boucle, S.; Bassit, L.; Ehteshami, M.; Schinazi, R. F. Clin. Liv. Dis. 2016,
20, 737.
>100
31
15
Guo, L.; Lin, X.; Liu, H.; Qiu, Z.; Shen, H.; Tang, G.; Wu, G.; Zhang, W.;
Zhu, W. PCT patent WO2013/144129, 2013.
16 Kappe, O. Tetrahedron 1993, 49, 6937.
>10
>100
17
Representative procedure for the decarboxyhalogenation - Synthesis of
Cl
H
>10
>100
> 100
4
65
26e
26a: Carboxylic acid 25a (346 mg, 1.0 mmol), sodium iodide (750 mg, 5.0
mmol) and sodium carbonate (106 mg, 1.0 mmol) were combined in a
solution of water (8 mL) and methanol (8 mL). Stirring and sonication gave a
clear, colorless solution to which was added Oxone (431 mg, 0.7 mmol). The
solution was protected from light and stirred at room temperature for 20 min.
After addition of a saturated solution of sodium thiosulfate (50 mL), the
mixture was extracted with ethyl acetate (3 x 25 mL); the combined organic
phases were dried over sodium sulfate and concentrated under vacuum.
Purification by flash chromatography on silica gel (hexanes/ethyl acetate:
8/2) gave 26a (290 mg, 68% yield) as a yellow solid. 1H NMR (400 MHz,
CDCl3) δ 8.50 (d, J = 3.8 Hz, 1H, ArH), 8.21 (d, J = 7.2 Hz, 1H, ArH), 7.74
(t, J = 7.6 Hz, 1H, ArH), 7.51 – 7.40 (m, 1H, ArH), 7.40 – 7.30 (m, 1H, ArH),
7.20 – 7.08 (m, 1H, ArH), 6.99 (t, J = 8.2 Hz, 1H, ArH), 6.05 (s, 1H, CH),
2.26 (s, 3H, CH3). 13C NMR (101 MHz, CDCl3) δ 163.1, 160.6, 149.8, 147.8,
136.9, 133.5, 131.4, 131.3, 125.4, 121.3, 117.1, 116.8, 114.8, 114.6, 63.4,
24.7. HRMS (ESI) calcd. for C16H12ClFIN3 [M+H]+ : 427.9827; Found:
427.9814.
Cl
Br
Cl
Br
Cl
H
H
>10
7.1
>100
>100
>100
>100
80
>100
80
>100
33
≥100
>100
45
26f
26g
26h
26i
I
I
I
4.0 ± 1.1
5.7
>100
75
11
33
>100
>100
H
>10
>100
>100
28a
Cl
>10
>100
28
69
18
28b
HAP-
12
-
-
-
-
-
-
-
-
-
-
-
-
0.5 ± 0.3
0.3 ± 0.02
0.04 ± 0.03
>100
>100
>10
>100
28
>100
69
>100
18
GLS4
3TC
>100
>100
>100
18 Zych, A. J.; Wang, H.-J.; Sakwa, S. A. Tetrahedron Lett. 2010, 51, 5103.
ND: not determined
19
Representative procedures for the Buchwald–Hartwig amidation
–
Synthesis of compound 28a: Protected-5-iodo-dihydropyrimidines 27a (56
mg, 0.10 mmol), acetamide (122 mg, 0.50 mmol), tris(dibenzylideneacetone)
dipalladium (10 mg, 0.01 mmol), Xantphos (17 mg, 0.03 mmol), potassium
In conclusion, we have synthesized and evaluated more than
thirty CAE analogs of HAP-12 and GLS4. Among them, we