Y. Suhara et al. / Bioorg. Med. Chem. Lett. 12 (2002) 3255–3258
3257
Next, we examined ability of 2 and 2a–c to induce dif-
ferentiation of HL-60 cells, which are human promy-
elocytic leukemia cells, by NBT reduction method.22 As
shown in Table 1, the potencies of these compounds in
inducingHL-60 cell differentiation were markedly
higher than that of 1 (ca. 11ꢁ38 times more potent than
1), and the rank order of the potency was
2a>2c>2b>2. These data indicate that the 2a-sub-
stituted ‘double side chain’ 1a,25-dihydroxyvitamin D3
analogues (2a–c) have a considerably potent ability to
activate VDR like 2 as compared to the natural
hormone 1.23
10. Binderup, L.; Latini, S.; Binderup, E.; Bretting, C.; Cal-
verley, M.; Hansen, K. Biochem. Pharmacol. 1991, 42, 1569.
11. Fujishima, T.; Liu, Z.; Miura, D.; Chokki, M.; Ishizuka,
S.; Konno, K.; Takayama, H. Bioorg. Med. Chem. Lett. 1998,
8, 2145.
12. Suhara, Y.; Kittaka, A.; Nihei, K.; Fujishima, T.; Konno,
K.; Takayama, H. Abstracts of Papers, 53th Meetingof the
Vitamin Society of Japan, Awajishima, Hyogo, May 24–25,
2001, Vitamins 2001, 75, 197.
13. (a) Norman, A. W.; Manchand, P. S.; Uskokovic, M. R.;
Okamura, W. H.; Takeuchi, J. A.; Bishop, J. E.; Hisatake, J.;
Koeffler, H. P.; Peleg, S. J. Med. Chem. 2000, 43, 2719. (b)
Uskokovic, M. R.; Manchand, P. S.; Peleg, S.; Norman, A. W.
In Vitamin D: Chemistry, Biology, and Clinical Applications of
the Steroid Hormone; Norman, A. W., Bouillon, R., Tho-
masset, M. Eds.; University of California: Riverside, 1997; p
19. (c) Kurek-Tyrlik, A.; Makaev, F. Z.; Wicha, J.; Zha-
binskii, V.; Calverley, M. J. In Vitamin D: Chemistry, Biology,
and Clinical Applications of the Steroid Hormone; Norman, A.
W., Bouillon, R., Thomasset, M. Eds.; University of Cali-
fornia: Riverside, 1997; p 30.
In conclusion, we have introduced the positive motifs,
2a-methyl, 2a-(3-hydroxypropyl), and 2a-(3-hydro-
xypropoxy) substituents, into the biologically active
vitamin D3 analogue of ‘double side chain’, 2. The A-
ringprecursor of the 2 a-methyl derivative 2a5 was ste-
reoselectively synthesized from chiral template 3. Bind-
ingaffinity for bovine thymus VDR was evaluated and
found that 2a–c possess higher potency of binding than
that of the parent analogue 2. All three analogues 2a–c
have shown considerably higher differentiation activity
in HL-60 cells than the natural hormone 1. Further
detailed studies of biological activities of these
analogues are currently in progress in our laboratories.
14. Trost, B. M.; Dumas, J.; Villa, M. J. Am. Chem. Soc.
1992, 114, 9836.
15. Wiggins, L. S. Methods Carbohydr. Chem. 1963, 2, 188.
16. Pougny, J.-R.; Sinay, P. J. Chem. Res., Miniprint 1982,
0186.
17. To
a solution of bromomethyltriphenylphosphonium
bromide (596 mg, 1.37 mmol) in THF (15 mL) was added
1.0 M NaHMDS in THF solution (1.28 mL) at À78 ꢀC under
Ar. The mixture was stirred at À78 ꢀC for 1 h, and then the
ketone13 with the ‘double side chain’ (100 mg, 0.273 mmol) in
THF (5 mL) was added dropwise at À78 ꢀC. After the reaction
mixture was stirred at room temperature, the solution was
diluted with hexane (100 mL). The resultingsolution was fil-
tered through Celite followed by concentration of the filtrate.
The resin was purified with silica gel column chromatography
(hexane/EtOAc 5:1) to give bromoolefin 12 (82 mg, 68% yield)
as a colorless oil. Data for bromoolefin 12: [a]2D0 +41.4 (c 0.77,
CHCl3); 1H NMR (400 MHz, CDCl3) d 0.56 (3H, s), 1.22
(12H, s), 1.81–1.87 (1H, m), 1.92–1.94 (1H, m), 1.96–2.01 (1H,
m), 2.86–2.89 (1H, m), 5.65 (1H, t, J=1.6 Hz) ; 13C NMR
(100 MHz, CDCl3) d 12.0, 19.8, 20.1, 21.9, 22.6, 27.0, 29.25,
29.36, 29.39, 30.9, 31.1, 31.3, 39.3, 39.6, 44.4, 44.5, 45.6, 52.3,
55.9, 71.0, 97.5, 145.1; HREIMS calcd for C24H43O279Br
(M+) 442.2447, found 442.2447.
Acknowledgements
The authors thank Misses J. Shimode and M. Kitsu-
kawa (Teikyo University) for spectroscopic measure-
ments. This work has been supported in parts by
Grants-in-Aid from the Ministry of Education, Science,
Sports and Culture of Japan.
References and Notes
18. Enyne 11 (20.0 mg, 54.3 mmol) and vinyl bromide 12 (25.0
mg, 56.5 mmol) were dissolved in TEA/toluene (3:1, 2.0 mL),
and tetrakis(triphenylphosphine)-palladium (0) (100 mg, 86.5
mmol) was added. After 15 min of stirringat room tempera-
ture, the resultant yellow solution was refluxed for 2 h. The
reaction mixture was filtered through a silica gel pad. Con-
centration followed by preparative thin-layer chromatography
on silica gel (20% EtOAc in hexane) gave crude 13 as white
solids, which was used in the next step without further pur-
ification. To a cooled (0 ꢀC) and stirred solution of crude 13 in
THF (2.0 mL) was added 1.0 M tetrabutylammonium fluoride
in THF (0.5 mL). After 1 h stirringat 0 ꢀC, the reaction mix-
ture was allowed to warm to room temperature and stirred for
further 12 h. The resultant solution was diluted with EtOAc
(10 mL), and washed with brine (3Â1 mL). The aqueous layer
was extracted with EtOAc (3Â2 mL), and the combined
organic layer was dried over Na2SO4. Filtration and con-
centration followed by preparative thin-layer chromatography
on silica gel (10% MeOH in CH2Cl2) gave (2.2 mg, 12% yield
in two steps) of 2a-methyl analogue 2a as a white solid. Data
for 2a-methyl analogue 2a: [a]2D0 +32.4 (c, 0.034, CHCl3); UV
(EtOH) lmax 267 nm, lmin 227 nm; 1H NMR (400 MHz,
CDCl3) d 0.53 (3H, s), 1.22 (12H, s), 1.90–1.94 (2H, m), 2.23
(1H, dd, J=8.0, 13.7 Hz), 2.67 (1H, dd, J=4.1, 13.7 Hz), 2.84
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