December 2009
1433
Table 1. Relative Binding Affinity for VDR and Osteocalcin Promoter 10) Sawada D., Katayama T., Tsukuda Y., Saito N., Takano M., Saito H.,
Transactivation Activity in HOS Cells of 1, 14-epi-pre1, and New Com-
pounds
Takagi K., Ochiai E., Ishizuka S., Takenouchi K., Kittaka A., Bioorg.
Med. Chem. Lett., 19, 5397—5400 (2009), and references cited
therein.
Osteocalcin
transactivation activity
(ED50 (nM))
11) Shimizu M., Iwasaki Y., Yamada S., Tetrahedron Lett., 40, 1697—
1700 (1999).
12) Ohno A., Shimizu M., Yamada S., Chem. Pharm. Bull., 50, 475—483
(2002).
13) Saito N., Masuda M., Saito H., Takenouchi K., Ishizuka S., Namekawa
J., Takimoto-Kamimura M., Kittaka A., Synthesis, 2005, 2533—2543
(2005).
14) Ohtani I., Kusumi T., Kashman Y., Kakisawa H., J. Am. Chem. Soc.,
113, 4092—4096 (1991).
Compound
VDRa,b,c)
1
2aR
14-epi-pre1
3aR
3bR
100b,c)
2.9c)
0.03
0.22
0.46
20.5
40.5
0.5b)
0.1b)
0.95b)
15) Enynes 7a and 7b were used as diastereomixtures, since it was difficult
to separate each diastereomer at this stage without HPLC, respectively.
16) Trost B. M., Dumas J., Villa M., J. Am. Chem. Soc., 114, 9836—9845
(1992).
a) The potency of 1 is normalized to 100. b) Chick intesinal VDR was used. c)
Human VDR was used.
17) Deprotection of the TES group was essential for easy separation of the
coupled compound from the starting materials in the next reaction
(Chart 5).
18) Spectroscopic data of 14-epi-1a,25(OH)2preD3, see: Maynard D. F.,
Trankle W. G., Norman A. W., Okamura W. H., J. Med. Chem., 37,
2387—2393 (1994).
19) In 1H-NMR analyses, there were no 6-s-trans signals. Therefore, we
estimated the ratio of the two isomers 6-cis/6-s-trans as 95/ꢂ5. Spec-
troscopic data for 3aR: [a]D19 ꢁ7.8 (cꢀ0.023, CHCl3); UV (EtOH)
l
max 250.0 nm, lmin 228.0 nm; IR (neat) 3379, 1466, 1377, 1215 cmꢁ1
;
1H-NMR (400 MHz, CDCl3) d: 0.86—1.56 (m, 33H), 3.30—3.34 (m,
3H), 3.62—3.66 (m, 1H), 4.03—4.10 (m, 1H), 4.22 (d, Jꢀ5.6 Hz, 2H),
4.49 (d, Jꢀ2.4 Hz, 1H), 5.73 (t, Jꢀ3.7 Hz, 1H), 5.92 (d, Jꢀ5.9 Hz,
2H); 13C-NMR (100 MHz, CDCl3) d: 13.9, 19.5, 19.8, 20.5, 21.7,
29.0, 29.3, 29.4, 29.4, 29.8, 34.1, 44.3, 44.4, 45.6, 51.6, 53.4, 68.3,
69.5, 71.1, 71.6, 77.1, 77.2, 77.5, 100.5, 119.5, 126.0, 132.3; EI-HR-
MS Calcd for C27H44O4 [Mꢃ+Na]ꢃ 455.3132, Found 455.3139.
1a,25(OH)2preD3 (14-epi-pre1) for the first time. We evalu-
ated VDR binding affinity and osteocalcin promoter transac-
tivation activity in HOS cells. Unfortunately, the biological
activities were not improved by 4-substitution; however, the
information on new compounds will be useful to create novel
potential ligands for VDR.
20) In 1H-NMR analyses, there were no 6-s-trans signals. Therefore, we
estimated the ratio of the two isomers 6-cis/6-s-trans as 95/ꢂ5. Spec-
troscopic data for 3bR: [a]D19 ꢁ142.5 (cꢀ0.07, CHCl3); UV (EtOH)
lmax 250.5 nm, lmin 228.0 nm; IR (neat) 3387, 1466, 1377, 1240,
Acknowledgements We are grateful to Ms. Junko Shimode and Ms.
Ayako Kawaji (Teikyo University) for the spectroscopic measurements. This
work was supported in part by a Grant-in-Aid from the Ministry of Educa-
tion, Culture, Sports, Science and Technology, Japan (to D.S.) and by a
Grant-in-Aid from the Japan Society for the Promotion of Science (to A.K.).
1213 cmꢁ1 1H-NMR (400 MHz, CDCl3) d: 0.89 (s, 3H), 0.93 (d,
;
Jꢀ6.4 Hz, 3H), 1.04—1.08 (m, 1H), 1.18—2.08 (m, 28H), 2.16 (s,
2H), 3.47 (s, 3H), 3.67 (d, Jꢀ6.4 Hz, 1H), 4.03—4.12 (m, 2H), 5.69 (s,
1H), 5.86 (d, Jꢀ12.5 Hz, 1H), 5.98 (d, Jꢀ12.5 Hz, 1H); 13C-NMR
(100 MHz, CDCl3) d: 17.2, 19.6, 19.8, 21.6, 21.9, 23.1, 29.1, 29.6,
33.9, 34.1, 35.8, 40.9, 44.4, 45.8, 52.0, 54.8, 56.4, 59.9, 65.6, 66.5,
69.1, 69.9, 71.0, 126.1, 127.4, 133.2, 137.1, 138.0; EI-HR-MS Calcd
for C27H44O4 [MꢃNa]ꢃ 469.3288, Found 469.3274.
References and Notes
1) For new vitamin D analogs (drugs and drug candidates), see: Posner G.
H., Kahraman M., “Vitamin D,” 2nd ed., Elsevier Academic Press,
New York, 2005, pp. 1405—1422.
2) For regulation of immune responses, see: Adorini L., “Vitamin D,” 2nd
ed., Elsevier Academic Press, New York, 2005, pp. 631—648.
3) For mechanism of action, see: Kato S., Fujiki R., Kitagawa H., “Vita-
min D,” 2nd ed., Elsevier Academic Press, New York, 2005, pp. 305—
312.
4) DeLuca H. F., Nutrition Rev., 66 (Suppl. 2), S73—S87 (2008).
5) Brown A. J., Slatopolsky E., Mol. Aspects Med., 29, 433—452 (2008).
6) Bouillon R., Okamura W. H., Norman A. W., Endocr. Rev., 16, 200—
257 (1995).
7) Zhu G. D., Okamura W. H., Chem. Rev., 95, 1877—1952 (1995).
8) Ettinger R. A., DeLuca H. F., Adv. Drug Res., 28, 269—312 (1996).
9) Kubodera N., Heterocycles, in press, DOI: 10.3987/REV-09-SR(S)3.
21) Spectroscopic data for 2aR: [a]D21 ꢁ87.3 (cꢀ0.01, CHCl3); UV
(EtOH) lmax 266.0 nm, lmin 212.5 nm; IR (neat) 3383, 1215 cmꢁ1; 1H-
NMR (400 MHz, CDCl3) d: 0.93 (d, Jꢀ6.4 Hz, 3H), 1.19—2.17 (m,
31H), 2.87 (d, Jꢀ12.0 Hz, 1H), 3.72 (ddd, Jꢀ7.1, 7.1, 14.2 Hz, 2H),
4.19 (s, 1H), 4.27 (s, 1H), 4.42 (d, Jꢀ6.1 Hz, 1H), 5.0 (d, Jꢀ2.0 Hz,
1H), 5.4 (s, 1H), 6.02 (d, Jꢀ11.7 Hz, 1H), 6.66 (d, Jꢀ11.7 Hz, 1H);
13C-NMR (100 MHz, CDCl3) d: 17.5, 19.8, 20.0, 20.1, 20.8, 22.0,
23.3, 29.1, 29.3, 29.6, 29.7, 30.1, 34.3, 36.1, 38.2, 41.5, 44.7, 45.9,
51.8, 51.9, 52.6, 69.9, 71.5, 77.5, 126.0, 126.3, 134.7, 138.7; EI-HR-
MS Calcd for C27H44O4 [MꢃNa]ꢃ 455.3132, Found 455.3113.