562
A. Duran et al. / Il Farmaco 57 (2002) 559–564
Table 3
Tested panels and cell lines
benzoyl), 8.07 (1H, d, C5H of napht.), 8.13 (2H, d, C4H
and C8H of napht.), 8.42 (1H, s, C1H of napht.), 13.86
(1H, s, NH).
Panel
Cell line
Leukemia
CCRF-CEM, HL-60(TB), K-562. MOLT-4,
RPMI-8226, SR
3.1.4. General procedure for the preparation of
1,4-dihydro-1-(2,6-dichlorobenzoyl)-3-(3-hydroxy-
2-naphthyl)-4-ethyl-5H-1,2,4-triazoline-5-thione (3a2)
2,6-Dichlorobenzoyl chloride was added to a solution
of 2a in acelone and refluxed at 120–140 °C for 2 h.
The precipitated product was filtered and recrystallized
from ethanol [4]. C21H15Cl2N5O2S, 96%, m.p. 210 °C.
Elemental analysis, (Calc./Found%): 56.77/56.77 (C),
3.40/3.31 (H), 9.46/9.33 (N), 7.21/7.22 (S).1H NMR (l,
ppm): 1.15 (3H, t, CH3), 4.10 (2H q, CH2), 7.37 (2H, t,
m-protons to chlorines on benzoyl and C4H of napht.),
7.53 (1H, t, C7H of napht.), 7.64–7.73 (3H, m, o-pro-
tons to chlorines on benzoyl and CgH of napht.), 7.82
(1H, d, C5H of naphl.), 7.94 (1H; d, C8H of napht.),
8.06 (1H, s, C1H of naphl.), 10.93 (1H, s, OH).
Non-small cell
lung cancer
Colon cancer
EKVX, HOP-62, HOP-92, NCI-H226,
NCI-H23, NCI-H322M, NCI-H460, NCI-H522
HCC-2998, HCT-116, HCT-15, HT29, KM12,
SW-620
SF-268, SF-295. SF-539, U251
LOX fMVI, M14, SK-MEL-2, SK-MEL-28,
SK-MEL-5, UACC-257
IGROVI, OVCAR-3, OVCAR-4, OVCAR-5,
OVCAR-8, SK-OV-3
786-0, A498, ACHN, CAKI-1, RXF 393,
TK-10, UO-31
CNS cancer
Melanoma
Ovarian cancer
Renal cancer
Prostate cancer PC-3, DU-145
Breast cancer MCF7, NCI/ADR-RES; MDA-MB-231/ATCC,
HS 578T, MDA-MB-435, MDA-N
3.2. Anti-tumor acti6ity screen
The NCl’s in vitro anti-tumor screen consisled of 52
human tumor cell lines (Table 3) against which 3a was
tested at a minimum of five concentrations at 10-fold
dilutions. A 48 h continuous drug exposure protocol
was used, and a sulforhodamine B (SRB) protein assay
was used lo estimate cell viability or growth. The
measured effect of the 3a on a cell line was currently
calculated according lo one or the other of the follo-
wing two expressions:
Fig. 3. Dose–response curves of selected six cell lines.
If (Mean Odtest−Mean Odtzero)]0
PG=100×(Mean Odtest−Mean Odtzero
/(Mean Odctrl−Mean Odtzero
)
)
)
against six cell lines for anti-tumor activity screen. Out
of 52 cell lines studied, only six were presented here.
The first column describes the subpanel and cell line
involved. The second column lists the Mean Odtzero and
Mean Odctrl. The third column lists the mean optical
densities (Mean Odtest) for each of five different concen-
trations and their calculaled percent growth in
paranthesis. Each concentration was expressed as the
log10 (molar or mg/ml). The next column lists the res-
ponse parameters GI50 (%50 growth inhibition), TGI
(total growth inhibition) and LC50 (lethal concentra-
tion), these parameters were interpolated values repre-
senting the concentrations at which the PG was +50, 0
and −50, respectively. In the case which these response
parameters can not be obtained by interpolation, the
value given for each response parameter is the highest
concentration tested and is preceded by a ‘\’ sign. The
last column lists the log10 of the values in fourth
column and these values were calculated from the
curves in Fig. 3. The values extending to the smaller
If (Mean Odtest−Mean Odtzero)B0
PG=100×(Mean Odtest−Mean Odtzero
/Mean Odtzero
Mean Odtzero is the average of optical density measure-
ments SRB-derived color just before exposure of cells
to the test compound. Mean Odtest is the average of
optical density measurements SRB-derived color after
48 h exposure of cells to the test compound. MeanOdctrl
is the average of optical density measurements SRB-
derived color after 48 h with no exposure of cells to the
test compound. PG is the percentage growth, the calcu-
lated measurement of effect.
4. Results and discussion
Table 3 shows all tested human tumor cell lines.
Table 4 represents the experimental data collected