Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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1
POI~YTHIOPHENE ANTI-TUMOR AGENTS
Field of the Invention
. The present invention relates to compositions and a
method for treating a patient having a tumor. -More
specifically, the present invention relates to the
treatment of such patients with an effective amount of a
polythiophene derivative.
Government Rights
This invention was made with United States Government
support under Grant No. 5 UO1 CA50743-02, awarded by the--
National Cancer Institute. The United States Government
has certain rights in the invention. This invention was
made with the support of the Republic of China under
National Science Council Project numbers 160061 45202,
45208 and 30602 and Ministry of Economic Affairs Project
Numbers 31X5110, 33A5100, 34B3300, 35M3100, 37A2100 and
13B12200. The Republic of China has certain rights in the
invention.
Backctround and Summarv of the Invention _
The control and cure of cancer represents one of our
most challenging health problems.-Tie treatment of cancer
can be approached by several modes of therapy including
surgery, radiation, chemotherapy.or a combination of any of
these treatments. Chemotherapy continues to be an
indispensable therapy for inoperable or metastatic forms of
the disease.
The selection of natural compounds, or=the synthesis
of new compounds having effective anticancer activity is
" complicated by the still limited knowledge of cancer cell
biology,and biochemistry. Therefore, development of new
effective anti-tumor agents will remain heavily dependent
- 35 on screening compounds to discover novel compounds having
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cytotoxic activity. Preferably , such compounds exhibit
enhanced cytotoxicity against tumor cells relative to their
cytotoxicity to normal cells.
Natural products have a history of providing,novel,
clinically useful anticancer drugs. Many active natural
products have also served as prototypes for the development
of new analogs of clinical and preclinical importance. Some
specific examples are-the Vinca alkaloids (vincristine,
vinblastine, vindesine and vinorelbine), podophyllotoxins
etoposide and teniposide), taxanes (taxol, taxotere),
camptothecins (10-hydroxycamptothecin-, 9-
dimethylaminomethylcamptothecin, 9-aminocamptothecin and
CPT-li), homoharringtonine, adriamycin, daunomycin,
bleomycin, mitomycin, idamycin, plicamycin and
dactiri~nycin. It is clear that natural products will
continue to be important sources of novel anticancer
agents. However, key to the success of novel antitumor
drug development programs is the initial identification of
potential antitumor agents.
The mouse L1210 leukemia cell line was initially the
preferred model system used for screening natural compounds
for antitumor activity. However, the P388 murine leukemia ,,
system was found to be more sensitive and predictive than
L1210 leukemia system, and has been used as primary screen
during the past decade. Systematic screening for compounds
exhibiting toxicity to these two leukemia cell lines has
resulted in the isolation of a large number of active
natural products. However, the anticancer activities of
these compounds were predominantly in leukemia, lymphoma
and a few rare tumors. Low clinical efficacy, or the lack
of clinical efficacy of known chemotherapeutics against
slower growing solid tumors, is a serious concern.
It has been recognized that the use of a single
antileukemia screening system could bias the end results -
and lead to the isolation of compounds only active in the
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3
treatment of fast growing tumors. In addition, the use of
a single antileukemia screening system may not detect novel
compounds with high specificities for particular cell
lines. It is also likely that many novel compounds with
possible anti tumor activity have remained undetected by
the less sensitive in vivo models due to the low
concentrations at which many active natural products occur.
Considering the diversity of tumors in terms of cell
type, morphology, growth rate and other cellular
l0 characteristics, the U.S. National Cancer Institute (NCI)
has developed a "disease-oriented" approach to antitumor
activity screening (M.R. Boyd, in "Principle of Practice of
Oncology" J.T. Devita, S. Hellman, S.A. Rosenberg (Eds.)
Vol. 3, PPO Update, No. 10, 1989). This in vitro
- prescreening system is based on the measurement of
antitumor cytotoxicity against human tumor cell line panels
consisting of approximately 60 cell lines of major human
- tumors (including leukemia and slower growing tumor cells
such as lung, colon, breast, skin, kidney, etc.). The most
important advantage of the new in vitro screening panels is
the opportunity to identify compounds that are selectively
more cytotoxic to cells of slowly growing solid tumors than
to-rapidly growing leukemia cells.
Thiophenes are sulfur containing heterocyclic
compounds that are distributed widely among the species of
the Asteraceae (Compositaie) family, including many species
with known_medicinal uses. The natural thiophene compounds
are thought to play an important role in the chemical
defense of plants against herbivorous insects and other
pests. Natural-thiophenes have been previously described
as having cytotoxic activities upon exposure to long
" wavelength ultraviolet light. Photochemical studies
suggest that thiophene phototoxicity is based primarily on
the production of toxic singlet oxygen by a type II
photodynamic process. However, polythiophene compounds
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4 "
also exhibit cy~otoxic activity in the absence of light
activation. More particularly, we have demonstrated that
the polythiophene derivatives of the present invention are
effective antitumor agents.
In accordance with this invention there is-provided a
method for the treatment of cancer which utilizes.
polythiophene compounds of the general formula:
1o R1
_ 2 I ( 3
_ S Jn
wherein n is 0, 1 or 2, Rl is H, CHO, CH20H or CH2NH2, and
R2 and R3 are independently optionally substituted 2- or 3-
thienyl. Further in accordance with this invention there
are provided novel cytotoxic compounds of the above formula
and chemotherapeutic pharmaceutical compositions containing
said compounds in anti-tumor effective amounts.
Additional objects, features, and advantages of the
invention will become apparent to those skilled in the art
upon consideration of the followingwdetailed description of
preferred embodiments exemplifying the best mode of the
invention as presently perceived.
_ Detailed Description of the Invention
The present invention is directed to polythiophene
compounds, their pharmaceutical compositions_and methods
utilizing such compounds/co~npositions for treating patients
having tumors. The polythiophene compounds are effective
antitumor.-agents against slow growing tumors. Generally---
they have been found to exhibit high selective cytotoxicity
3 5 for individual tumor cell lines. -
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The compounds of the present invention are
polythiophene compounds of the formula:
5 _ R1 - -
R2 ~ ~ R3
~n
- -
wherein w is 0, 1 or 2,
R1 is H, CH20H, CHO, CH2NH2,
R2 and R3 are independently selected from the group
consisting of 2-thienyl, 3-thienyl, mono- or di-
substituted 2-thienyl, or mono- or di- substituted 3-
thienyl, wherein the thienyl substituents are selected from
the group consisting of cyano, chloro, bromo, iodo, C1-C7
alkyl or haloalkyl, C1-C~ alkenyl or haloalkenyl, C1-C4
alkanoyloxy methyl, CH20R4, CORS, CH2NR6R7, CH(OR4)R8, _
CH=CRgRlo, CH=NR11, CH2SC(NH)NH2 and C=CR12 wherein
R4 is H, CO(CH2)2C02H, (CH2)20CH3, Cl-C4 alkyl, COC1-C17
alkyl, or tetrahydropyranyl;
RS is H or C1-C7 alkyl;
R6 and R7 are independently H, C1-C4 alkyl, or mono- or
di- hydroxyC2-C4 alkyl;
R8 is C1-C7 alkyl, or C1-C7 alkenyl;
R9 and Rlo are independently H, C1-C7 alkyl, COOR5, CN,
CH(OR4)COORS, Br, CO-thienyl, COC6H40H(p);
R11 is NHR4 flr ORS;
3 0 Rl2 is COORS , CH ( OR4 ) CH20R13 or CH ( OCOC 1-C4
alkyl) CH20R5; -
R13 is H, COCH2CH2COOH, or COCl-C1~ alkyl;
cyclodextrin complexes of such compound and
when--R2 or R3 is thienyl substituted with CH2NR6R~, the
pharmaceutically acceptable salt of the compound
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6
represented thereby; with the proviso, that
when R1 is H, R2 is selected from the group
consisting of 3-thienyl, di- substituted 2-thienyl,
hydroxymethyl- or aminomethyl- substituted 2-thienyl,_ 3-
_ 5 formyl-2-thienyl and mono- or di- substituted 3-thienyl,
and R3 is selected from the group consisting of 3-thienyl,
di- substituted 2-thienyl, hydroxymethyl- or aminomethyl-
substituted 2-thienyl, mono- or di- substituted 3-thienyl
and formyl substituted 2-thienyl.
In one preferred embodiment of this invention there is
provided anti-tumor polythiophenes of the above formula
wherein n is 1, R1 is H, R2 is 3-thienyl or substituted 3-
thienyl and R3 is 2-thienyl or substituted 2-thienyl. Such
polythiophene compounds, especially those wherein R2 is 3-
thienyl and R3 is 2-thienyl optionally substituted with
CH2NH2, CH20H or CHO, exhibit cytotoxic selectivity against
transformed human cells.
Other preferred compounds in accordance with this
invention are those polythiophenes of the above formula
wherein n is 1, R1 is hydrogen, R2 is hydroxymethyl 2-
thienyl and R3 is 2-thienyl optionally substituted with
CH2NH2, CH20H or CHO.
Still another preferred group of polythiophene
compounds of this invention are those of the above formula
wherein R2 is optionally substituted 2-thienyl and R3 is 3-
or 4-substituted-2-thienyl wherein the substitutes are
selected from CH20H, CHO and CH2NH2.
Another group of polythiophene compounds within the
scope of this invention are compounds of the formula:
R1
R2 I S I R3
n
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7 -
wherein n is 0, 1 or 2,
R1 is H, CH20H, CHO, CH2NH2,
R2 is selected from the group consisting of 2-thienyl,
- 3-thienyl, mono- or di- substituted 2-thienyl, or mono- or
di- substituted 3-thienyl, and R3 is selected from the
group consisting of 3-thienyl, mono- or di- substituted 3-
thienyl and di- substituted 2-thienyl, wherein the thienyl
substituents are selected from the group consisting of
cyano,-chloro, bromo, iodo, C1-C7 alkyl or haloalkyl, C1-C7
alkenyl or haloalkenyl, C1-C4 alkanoyloxy methyl, CH20R4; -
CORS , CH2NR6R7 , CH ( OR4 ) Re , CH=CRgRlO , .._,CH=NR11, CH2 SC ( NH ) NH2
and C---CR12 wherein
R4 is H, CO(CH2)2C02H, (CH2)20CH3, C1-C4 alkyl, or COC1-
C17 alkyl ;
R5 is H or C1-C7 alkyl; _
R6 and R7 are independently H, Cl-C4 alkyl, or mono-.or
di- hydroxyC2-C4 alkyl;
R8 is C1-C7 alkyl, or C1-C~ alkenyl; __
R9 and R10 are independently H, ~1-C~ alkyl, COORS, CN,
CH(OR4)COORS, Br, CO-thienyl, COC6H40H(p); _
R11-is NHR4 or ORS;
R12 is COORS, CH(OR4}CH20R13 or CH(OCOC1-C4
_ __ _. alkyl) CH2QR5;
Rl3 is H, COCH2CH2COOH, or COC1-C17 alkyl;
_ 25 cyclodextrin complexes of such~compound and
when R2 or R3 is thienyl-substituted with CH2NR6R~, the _
pharmaceutically acceptable salt of the compound
represented thereby.
The polythiophene compounds of this invention are
readily formulated into pharmaceutical compositions, also
within the scope of this invention, for use in the
' presently described method for treatment of patients having
tumors. In one preferred embodiment of this invention, the
- pharmaceutical composition comprises an anti-tumor effec
_ 35 tive amount of a polythiophene compound of the formula:
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R1
2 ~ ( ~
S~n
wherein n is 0, 1 or 2,
R1 is H, CH20H, CHO, CH2NH2,
R2 and R3 are independently selected from the group
consisting of 2-thienyl, 3-thienyl, mono- or di-
substituted 2-thienyl, or mono- or di- substituted 3=
thienyl, wherein the thienyl substituents are selected from
the group consisting of cyano, chloro, bromo, iodo, C1-C7
alkyl or haloalkyl, C1-C7 alkenyl or haloalkenyl, C1-C4
alkanoyloxy methyl, CH20R4, CORS, CH2NR6R~, CH(OR4)R8,
CH=CRgRlo, CH=NR11, CH2SC(NH)NH2 and C=CR12 wherein
R4 is H, CO(CH2)2C02H, (CH2)20CH3, or COC1-C1~ alkyl;
RS is H or C1-C7 alkyl;
R6 and R7 are independently H, C1-C4 alkyl, or mono- or
di- hydroxyC2-C4 alkyl;
R8 is C1-C7 alkyl, or C1-C7 alkenyl;
R9 and R1~ are independently H, C1-C7 alkyl, COORS, CN,
CH(OR4)COORS, Br, CO-thienyl, or COC6H40H(p);
R11 is NHR4 or ORS;
R12 is COORS, CH(OR4)CH20R13 or CH(OCOC1-C4
alkyl)CH20R5;
R13 is H, COCH2CH2COOH,~ or COC1-Cl~ alkyl;
cyclodextrin complexes of such compound and
when R2 or R3 is thienyl substituted with CH2NR6R~, the
pharmaceutically acceptable salt of the compound
represented thereby; with the proviso, that
when R1-is H, R2 is selected from the group
consisting of 2-thienyl, 3-thienyl, mono- or di-
substituted 2-thienyl, or mono- or di- substituted 3-
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thienyl, amd R3 is selected from the group consisting of 3-
thienyl, mono- or di- substituted 2-thienyl, or mono- or
di- substituted 3-thienyl,
and a pharmaceutically acceptable carrier.
Another pharmaceutical composition within the scope of
this invention comprises an anti-tumor effective amount of
compound of the formula:
R
io ( ~ - . .
R2_ R3
n
- 15
wherein n is 0, 1 or 2,
Rl is H; CH20H, CHO, CH2NH2,
R2 is selected from the group consisting of~2-thienyl,
3-thienyl, mono- or di- substituted 2-thienyl, or mono- or
20 di- substituted 3-thienyl, and R3 is selected from the
group consisting of 3-thienyl, mono--or di- substituted 3-
thienyl, di- substituted 2-thienyl wherein the thier~yl
substituents are selected from the group consisting of
cyano, chloro, bromo, iodo, Cl-C~ alkyl or haloalkyl, C1-C7
25 alkenyl or haloalkenyl, C1-C4 alkanoyloxy methyl, CH20R4,
CORS , CH2NR6R7 , CH ( OR4 ) R8 , CH=CR9Rl~ , CH=NRl1, CH2 SC ( NH ) NH2
and C~CRZ2 wherein
R4 is H, CO(CH2)2C02H, (CH2)20CH3, C~-C4 alkyl or COCl- _
_ CIA alkyl;
30 R5 is H or C1-C7 alkyl; --
R6 and R7 are independently H, C1-C~ alkyl, or mono- or
- _ di- hydroxyC2-C4 alkyl; -
R8 is C1-C7 alkyl, or Cl-C~ alkenyl;
- R9 and R1o are independently H, C1-C~ alkyl; COOR5, CN,
- 35. CH(ORQ)COORg, Br, CO-thienyl, or COC6H40H(p); _ -
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R11 is NHR4 or ORS;
R12 is COORS, CH(OR4)CH20R13 or CH(OCOC1-C4
alkyl)CH20R5;
- R13 is H, COCH2CH2COOH, or COC1-C17 alkyl;
5 cyclodextrin complexes of such compound and
when~R2 or R3 is thienyl substituted with CH2NR6R7, the
pharmaceutically acceptabl-a salt of the compound
represented thereby, and a pharmaceutically acceptable
carrier-:
ZO The present compounds are readily prepared using ar-t-- --
recognized chemical-synthesis procedures as exemplified
hereinbelow. The art is replete with descriptions of the
chemistry and synthesis of thiophene and polythiophene
compounds.
Tables 1, 2, 3 and 4 hereinbelow show several
polythiophene compounds that have been tested at the
National Cancer Institute. The growth inhibition profiles
of several polythiophenes are shown inlTables 5, 6 and 7.
The unusually strong activities against solid tumors are
predictive of significant therapeutic potency for the
treatment of cancer.
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Table -
~1
Compounds registered tested
and by
NCI
n = o
~R~
s L ~S J
~s
n
to
NSC Code R' -
R"
645279 CH20H H
649652 -- - - CH20Ts H
15 2fl-3017 COCH3 COCH3
649653 I I
649654 CH=CBr2 H
649656 CH=CHCOTh W--
~.
649657 CH=CHCOC6H40H(p) H
20 649658 CH2NHC6H5 H
649659 CH=NCH2CH(OH)CH20H H
647070 CH=C(CN)COOH H
645276 C=CCH(OH)CH20H H
645277 C=CCH(OAc)CH20H H
~
25 646271 CH=N-OH H
(syn)
646655 CH=N-OH H
(anti)
647073 CHO CH20H
647074 CHO C7H15
647450 CH20H C7H15
30 647451 COC6H13 H-
647452 _ CH20H CH20H
647453 CHO CH20Ac
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I2
Table 1 Continued
n = 0
R ~I ~ ~ . ~ _ ~ R
r S S~S
n
NSC Code R' R"
652869 CH20C0(CH2)2C02H CHO
542870 _,CH~OCO(CH2)2C02H CH20C0(CH2)2COZH
l5
THP: tetrahydropyranyl Ac: actyl
TS: tosyl _ Et: ethyl
TH: thienyl
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13 _.
_ _. Table 2
Compounds registered tested
and by
NCI
n = 1 .
.
R, I I C I ~R~
i
s La s mss.
~
n
to
NSC Code R' R~~
637393 CHO H
637394 CHO CHO
637388 CH20H H
645278 CH20Ac H
_.
639392 CH20Et H
659566 CH(OH)CH3 H
659567 COCH3 H
637389 CN H
637390 CN CN
637391 CH=CHCOOH H
-
637392 CH=C(COOH)2 H
64 62 67 CH=NNHC6H5 --
---- H
646268 CH=N-OH H
660642 CH2SC(NH2) H
- NH
646269 C=CC02Et H
646270 CH20H CH20H
646272 CH20(CH2)20CH3 H
647072 CH20H CH3
647454 CH3 - H
- 647455 CHO CH20H
64-9662 COC2Hg H
649663 CH(OH)C2-H5 H
-35 651690 CH20C0(CH2)2C02H H
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14
Table 2 Continued -
n = 1
R ~ ~ ~ ~ ~ ~ R.
S~S~S
n
NSC Code R' R"
652866 CH2NH2 H
660645 - CH2NH3 tartarate H
6898 CH2NH2 CHzOH
658110 CH2NH2 CH2NH2
659562 CH2NHCH3 H
658466 CH2NHCH2CH(OH)CH20H I
659563 CH2N(CH3)2 H
659561 CH2N(C2H5)3C1 H
659564 CH2N(CH3)2 CH2N(CH3)2
659565 CH2N(CH3)2 CH20H
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Table 3
Compounds registered and tested by NCI
n = 2
5
s L ~s J ~s
i o --
NSC Code R' R"
645273 -- CHO H
645274 CH20H H
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16
- Table
4
Compounds registered
and
tested
by
NCI
.r,
.,.,..
CHO
HOHZC
663565 I
S
S
- CH=OH
HoH2c
I
!
I
_
I
_
55356 S
S
CHO
I
(
I
S
65811I .
S
S
53~ S
S
S
CH=OH
I I ~ (
~ ~ _-
658112 S
g
S
...-_
CH=OH
I
S
S
S
658113 -
~O
S
68880
CHO
O~C-,
I1
~
I~
~
\
~
55887 S
S
S
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1'
Table 4 Continued
_ CHsOH _
OHC
5S8879 S S S
CHIOH
OHC
658876 - - S S S
- CH=OH
_- _ _____. OOH=C-~
658878 S S S
CH=OH
__ 15
638875 . HOH=C-~S ~S I I S
_ . CHO
_ I I I ~CHO
_ ~0 _ 663560 S S 5
CHO
i I I I I
S S S CH=OH
663562
CHZOH
25 I I I I (
S S g CH=OH
663561 -
._ CHZOH
OHC ~ - _ I I ~ I ~ ~O
3 0 666164 S S S
- cH,oH -
HOHzC--~ I I ~ ~ ~ ~=OH
666165 S S S
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Table 4 Continued
5~8~I--CHO
rJ
660643
_ .~ i 8 ~ I g I CHiOH _
6606;.e S
20
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Table 5: The growth f the human cancer cell
inhibition
o
lines
by polythiophene
Logio Glso~M~
Compd.
Cell Line
Lung Cancer
A549/ATCC -5.05 -5.18 -4.4-9
EKVX -4.61 -4.26
HOP-18 -4.98 -4.65 -5.63
HOP-62 -5.01 -4.62
NCI-H23 -4.15 -4.71 -4.26
NCI-H226 - -6.50 -6.93 -6.49 -7.61-
NCI-H322M -4.52 -4.53 -4.71 -4.37
NCI-H460 -5.87 -6.3Z -5.82- -4.86
NCI-H522 -4.02 -4.88 -4.-49
LXPL-529 -6.54 <-8.00 <-8.~ <-8.00
Colon Cancer
COLD-205 -5.44 -6.04 -6.36 -5.27
DLD-1 -6.50 -5.35 <-8.0Q -5.10
HCC-2998 -4.44 -4.61- -4.75
HCT-116 -5.34 -6.53 -7.20
HCT-15 -4.45 -4.30 -4.62
HT29 -5.07 -4.52 -4.61
KM20L2 -5.80 -6.6l -6.88 -6.31
SW-620 --5.33 -6.27
CNS Cancer
SF-268 -4.61 -4.06
SF-295 _-_4.42 -4.47 -4.74 '4.16
SNB-75 -6.41 <-8.00 -4.50
SNB-19 -4.73
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Table
5 Continued
Compd.
_ 5 Cell Line
CNS Cancer
U251 -6.34 -6.83 <-8.00
' XP-498 -5.43
10
Melanoma
- LOX IMVI -4.62
MALME-3M -4.49 - -4.l0
M-14 -4.40 -4.20
15 SK-MEL-2 -4.14 -4.51
SK-MEL-28 -4.05
UACC-257 -6.03 -6.99 -7.03 -7.33
UACC-62 < -8.00 -4.77 -8.00
20 Ovarian Cancer
IGROV-1 -7.37 -5.30 -7.22
OVCAR-3 -5.63 -7.59 -7.52
OVCAR-4 -4.51
OVCAR-5 -4.89 -6.65 -4.58 -6.86
OVCAR-8 -4.28 -4.27
SK-OV-3 -4.04 -4.06
Renal Cancer
786-0 -4--.49 -4.73 -4.36
A-498 < -8.00 -7.36 -7.36
w
ACHN -4.75 -4.29
CAKI-1 -6.65 -7.56 -7.?4 -7.86
RXF-393 -4.24 -7.50 -4.74 -4.78
RXF-63l -6.09 -7.55 <-8.00
SN12C -4.54 -4.32 -
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zl
- Table 5 Continued -
Compd.
Cell Line
Renal Cancer ---
TK-10 -6.63 -6.98 -6.92r6.28
UO-31 -4.41 -4.73 -4.38
__. GISO: molar concentration required for _-.
50% growth inhibition
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2 2 ._
Table 6: The growth inhibition the human cancer cell
of
lines by chemically modified a-polythiophene
derivatives
Logio GI50 M
Compd.
Cell Line
Lung Cancer
A54 9 /ATCC -5 . -----~-----
91
HOP-18 -5.51
HOP-62 -5.52
HOP-92 -4.09
NCI-H226 -7.60 -7.88 -5.76 -5.23
NCI-H322M -5.77
NCI-H460 -6.29 -7.80 -5.62
LXPL-529 -6.33 < -8.00 -5.95 -5.98
Colon Cancer
COLD-205 -6.22 -6.81
DLD-1 -4.74
HCT-116 -5.27 -7.82 -4.74
HCT-15 -4.19
HT29 -5.04
KM20L2 -6.15 -6.84 -4.39
KM12 -4.35 , -
SW-620 -5.04 -7.17
HCC-2998 -6.40 -5.74
CNS Cancer -
SNB-78 -4.44
SNB-75 -4.43 r 4:14
SNB-19 -4.72 -4.97
U251 -7.73 -6.09
XP-498 -7.52 -4.96
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WO 98I23269 PCT/US96/1.9118
23
- Table nued
6 Conti
Compd.
Cell Line
CNS Cancer
SF-295 . -4.43
Melanoma
UACC-257 -6.00 -7.85.72 __-4.97
UACC-62 -6.36 <-8.i.46
Ovarian Cancer-
IGROV-__1 -6.57 <-8.9.15 -5.95
OVCAR-3 -5.70 <-8.00
OVCAR-4 -4.46
OVCAR-5 -6 . -7 . y4 . -
06 68
OVCAR-8 -5.23 -4.l0
SK-OV-3 -5.80 -5.38
Renal Cancer
A-498 -7.71 <-8.i.93
CAKI-1 -6.63 <-8.e.88 - -6.21
-
RXF-631 <-8.00 r4.03-4.96
TK-1025p -6.29 -7.85.29 -6.05
RXF-393 -7.05
GISO: Concentration required for 50% growth inhibition
CA 02272275 1999-OS-18
WO 98I23269 - " PCT/US96I19118
24
~,.
Table 7: growth inhibition of the human solid cancer cell
lines by polythiophenes
LOGlO GIS~ (M) -
Compd. -
Cell Line -
Lung Cancer
HOP-62 -4.69 -4.70
___ HOP-92 -4.41 -5.03
NCI-H23 -4.62 > 5.00 -4.4l
NCI-H226 -7.-85 -6.59
NCI-H322M -4.79 -5.37
NCI-H460 -4.60 -7.41 -5.25
NCI-H522 -4.47
Colon Cancer
COLO-205 -6.04 -6.72 -5.23
HCC-2998 -6.01 -5.23 -6.50 -
HCT-116 -6.44 -6.9?
HCT-15 -4.23 > -5.00 -4.55
HT29 =- -4.86.63 -4.73 ----
KM12 -4~ 085. -4 . 78
00
SW-620 -4.32 -4.39
CNS Cancer
SF-268 -4.27 > -5.00
SF-295 -4.45 > -5.00 -4.47
SF-539 > -5.00 - -
SNB-19 -4.48 > -5.00
SNB-75- -4.51 -5:70
U251 -4.95.25 -4.16
CA 02272275 1999-OS-18
WO 98/23269 PCT/US96/19118
2 5 --
Table
7 Continued
- Compd.
Cell Line
Melanoma
MALME-3M -4.41 > -5.00 r 4.53
M14 -4~ 3~5. -4 . 29
00
IO SK-MEL-28 -4.44 > -5.00 -4.40
SK-MEL-5 -4.38 > -5.00 > -5.00
UACC-257 -6.72 -7.42 -.
UACC-62 -6.74
LOX1MV1 - > -5.00
SK-MEL-2 > -5.00
ovarian Cancer
IGROV1 -6.66 -6.84 -6.64
OVCAR-3 -6.80 -7.16
OVCAR-4 -4.60 -6.52
OVCAR-5 -6.38 -6.90
OVCAR-8 -4.24 > -5.00 -4.30
Renal Cancer - -.-_
786r0 -4.24 -4.28
A498 -6.97.80
CAKI-1 -7.64 -7.60
RXF-393 =4.69 > -5.00 -5.06
TK-10 -6.69 -7.10 -4.68
UO-31 ~ -4.19
SN12C - >.-5.00 -4.33
ACHN > -5:00 -4.25
_ Prostate Cancer
PC-3 -4295.00 -4.31 -
CA 02272275 1999-OS-18
WO 98l23269 PCT/US96119118
26
Table 7 Continued -
Cell Line -
Prostate Cancer
DU-145 - -4....18 > -5.00 -4.26
l0 Breast Cancer
MCF7 -6. y6. 86 - -6. 75
MCF7/ADR- > -5.00 -4.-2-4-
RES
MDA-MB- -- -4.55 > -5.00
231/ATC-C
MDA-N -6.30 -5.02 -5.27
T-47D -6.47 -6.65 -6.37
HS 578T -5.11 - . -4.2,8
MDA-MB- -5.36 -4.82
435
BT-549 > -5.00 -4.08
CA 02272275 1999-OS-18
WO 98/23269 PCT/US96/19118
27
The cytotoxic activity of the present polythiophene
compounds have been measured utilizing three different
assays or screens. The first screen measures the
cytotoxicity against a panel of sixty different human tumor
cell lines) This assay provides data regarding the general
cytotoxicity of an individual compound. In particular this
type-of assay is useful in identifying compounds which have
enhanced cytotoxic activity against slow growing tumors as
1o compared to fastergrowing tumor cells such as leukemia
tumor cell lines. The identification of such compounds is
critical since previously identified antitumor agents have
low cytotoxic activity against slower growing tumors. The
specificity of a compound for a limited number of tumor
cell lines also indicates that such a compound will likely
be less cytotoxic to normal cells. The specificity of a
cytotoxic compound for tumor cell lines relative to normal
- cells is an important characteristic of an effective
antitumor agent.
Antitumor cytotoxicity data for the National Cancer
Institute human tumor cell panels can also be expressed in
a graphic pattern (mean graph) to display differential cell
growth inhibition (K.D. Paull, R.H. Shoemaker, L. Hodes, A.
Monks, D.A. Scudiero, L. Rubinstein, J. Plowman and M.R.
Boyd,- J= Natl. Cancer Inst., 81, 1088, i989.) In the mean
graph, the arithmetic mean of the logarithm of the GISO (50%
growth inhibition), TGI (total growth inhibition) or LC5o
(50% lethal concentration) values is used as an anchor
point. Relative cytotoxicity is displayed by projecting
bars to the right or left of the mean, depending on whether
cell sensitivity to a test compound is more or less than
average. The length of mbar is indicative of differential
cytotoxicity against a specific type of tumor cells or
- tumor panels.
In a second assay, the cytotoxic selectivity is. assessed
- CA 02272275 1999-OS-18
WO 98/23269 PCT/US96/19118
28
by comparing-.cs~npound cytotoxicity against transformed
cells and normal cells. ICSO values were compared between
- treated TBE cells (ras-transformed human bronchial
epithelial cells) and NHBE cells (Normal human bronchial
epithelial cells). Cytotoxic effects on normal human
bronchial epithelial cells (NMBE) and ras-transformed human
bronchial epithelial cells (TBE) were measured by the cell
count (cell number) using Coulter Z.F. counter (Hialeah,
FL) carried out at Purdue University. The result is
expressed as GISO, concentration of drug at which cell
numbers are reduced to 50% of control cell culture (T.C.K.
Chan, C.-j. Chang, N.M. Koonchanok and R.L. Geahlen,
Biochem. Biophys. Res. Commun., 193, 1l52, 1993). The data
presented in Table 8 illustrates that polythiophenes
generally exhibit greater cytotoxicity for transformed
human cells in comparison to the normal human cells.
The antitumor cytotoxicity of the thiophene compounds
tested in the first two in vitro assays was measured by a
microculture assay using either
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide (MTT) or sulforhodamine B (SRB). [M.R. Boyd in
"Principles and_Practices of Oncology," V.T. DeVita, Jr.,
- CA 02272275 1999-OS-18
WO 98I23269 w - PCTIUS96/19118
29
Table 8: Selective cytotoxicity
against ras-ol7cogene
transformed human bronchial cells
epithelial
- GIso
(ug/ml)
I
I
I
I
I
I
Ci30
637393 S 0.03 O.Z
S
S
I I I
I i ~--cx,oH
s373as S 0.02 3.0
s
s
_
1 I I
f ( t
~_
_ _ 6s~a66 S o.ool o:ol
s
s
-
Q9561 ~ ~ 2
I ~
~
~
~2,o;;~s~~
s
S
5
_ ~ ~ I I
~ I
o ~O
C S
S
S
~~g4 0.01 OZ
-----._ _ I __
HOH:C I
I I
I
I
O=OH
_ S s 0.02 1.0
~0 5
2 0
HOHZC- ~ I
I ~C2io
_ Q70; 3 S 0.05 0.08 . ..
HOCI-1:I ( ~=oH
I I
2 5 6:7452 S S 0.03 0.6
c--~ I I ~ cxo
I I
Hors
=
6d7455 S S 0.Q020.1
S
H I ~ (
Ov_C~ S - ~ S C~i~NH~
S.
.02 .0
3 0 Q6898 . 0 3
H='~=y.. I S ~ y 5 ~ ' S I cH=NHi
658110 4.0 10
a3=oH - .
o~c ~ I I _ _
65EE79 S S 5 0.00: 6.2
CA 02272275 1999-OS-18
WO 98/23269 PCT/US96/19118
Table 8 Continued
CH=OH
HOHZC _
6S8878 S S S Q.002 8.0
CHO
OHC
65EE77 S S S 0.03 0.6
r ~_ CH=OH _
658876 OHC---.~S S . S 0.02 0.4
CH=OH'
658E75 HOH:C--~ ~ 0.002 9.7
S S S
1 _ _ 1 I _ t cHo
1 . f S S.
2 0 ~3 5 0.005 1.4
S I I S j~-CH~pH
IJ
6 " ' S - 0.Q02 - 3.6
30
CA 02272275 1999-OS-18
WO 98/23269 PCT/I1S96/19118
S. Hellman and S.A. Rosenberg (Eds.), Vol. 3, PPO Updates,
Number 10, 1989.] This assay has an advantage over in vivo
assay in that results are obtained within a week as opposed
to several months. The assay was carried out in 96-well
microtiter plates. The MTT assay is based on the
production of a dark blue formazan product by dehydrogenase
in the mitochondria of live tumor cells after exposure to
drug for 6 days [M. C. Alley, D.A. Scudiero, A. Monks, M.L.
Hursey, M.J. Czerwinski, D.L. Fine, B.J. Abbott, J.G. Mayo,
R.H. Shoemaker and M.R. Boyd, Cancer Res., 48, 589, 1988.]
Thus, only live cells are stained and can be measured at
570 nm. The SRB assay is based on the binding of the
anionic group to the basic amino acid residues of cellular
proteins after exposure of tumor cells to drug for 2 days
[P. Skehan, R. Storeng, D. Scudiero, A. Monks, J. McMahon,
D. Vistica, J.T. Warren, H. Bohesch, S. Kenney and M.R.
Boyd, J. Nat. Cancer Inst., 82, 1107, 1990.) Thus, the
total protein (viability) can be measured at 564 nm.-
Antitumor cytotoxicity is reported as GISO,~ effect drug dose
at which cell growth is retarded to 50% of control culture
of tumor cells. The active compounds are defined as those
compounds having GISO values that are less than 10-4 M or 10
I~g/ml.
Antitumoral activity of the present polythiophene
compounds has been confirmed by in vivo animal test data.
The in vivo data was derived from experiments in which
human tumors are transplanted into immune deficient mice
and allowed to grow for two~days prior to treatment_with a
polythiophene composition of this invention. See Example
48 and accompanying Tables 9 and 10. Data obtained from
studies using heterotransplanted tumors in immune deficient
- mice are recognized as well-correlated with the
effectiveness of.-these agents in clinical studies
- (Giovanella, B.C, et al. Cancer 52(7): 1146 (1983). -
CA 02272275 1999-OS-18
WO 98I23269 PCT/US96/19118
32
The present invention further provides pharmaceutical
formulations comprising an effective amount of a
polythiophene compound for treating a patient having a
tumor. As used herein, an effective amount-of the
polythiophene compound is defined as the amount of the
compound which, upon administration to a patient, inhibits
growth of tumor cells, kills malignant cells, reduces the
volume or size of the tumors or eliminates the tumor
entirely in the treated patient.
The effective amount to be administered to a patient is
typically based on body surface area, patient weight, and
patient condition. The interrelationship of dosages for
animals and humans (based on milligrams per meter squared
of body surface) is described by Freireich, E.J., et al.,
Cancer Chemother. Reb., 50 (4): 219 (1966). Body surface
area may be approximately determined from patient height
and weight (see e.g., Scientific Tables, Geigy
Pharmaceuticals, Ardley, New York, pages 537-538 (1970)).
An effective amount of the polythiophene compounds in the
present invention can range from about 5 mg/kg. to about
500 mg/kg, more preferably from about 5 mg/kg to about 250
mg/kg, and most preferably about 5 to about 150 mg/kg.
_ Effective doses will also vary, as recognized by those
skilled in the art, dependant on route of administration,
excipient usage and the possibility of co-usage with other
therapeutic treatments including other anti-tumor agents,
and radiation therapy.
The pharmaceutical formulation may be administered via
the parenteral route, including subcutaneously,
intraperitoneally, intramuscularly--and intravenously.
Examples of parenteral dosage forms include aqueous
solutions of the active agent, in a isotonic saline, 5%
glucose or other well-known pharmaceutically acceptable
liquid carrier.' In one preferred aspect of the present
embodiment, the polythiophene compound is dissolved in a
CA 02272275 1999-OS-18
WO 98/23269 - PCT/US96/19118
33 _
saline solution containing 5% of dimethyl sulfoxide and 10~
Cremphor EL (Sigma Chemical Company). Additional
solubi.lizing agents such as cyclodextrins, which form
_ J- specific, more soluble complexes with the present
polythiophene compounds, or other solubilizing agents well-
known to those familiar with the art, can be utilized as
pharmaceutical excipients for delivery of the polythiophene
compounds.
The present compound can also be formulated into dosage
l0 forms for other routes of administration utilizing well -~~-----
known methods. The pharmaceutical compositions can be
formulated, for example, in dosage forms for oral
administration in a capsule, a gel seal or a tablet.
Capsules may comprise any well-known pharmaceutically
acceptable material such as gelatin or cellulose
derivatives. Tablets may be formulated in accordance with.
conventional procedure by compressing mixtures of the
active polythiopene and solid carriers, and lubricants
well-known to those familiar with the art. Examples of
solid carriers include starch, sugar, bentonite. The
compounds of the present invention can also be administered
in a form of a hard shell tablet or capsule containing, for
- _ _, example, lactose or mannitol as a binder and a conventional
fillers and tableting agents.
The following examples are provided to illustrate various
embodiments of Applicants' invention, and are not intended
to in any way limit the scope of the invention as set forth
in this specification and appended claims.
Example 1 -
Synthesis of 5-formyl-5'-heptyl-2.2'-bithiophene
- (NSC code # 647074)
Dimethyl formamide (DMF) (8 ml) was stirred at 0~C for 10
- minutes, POC13 (1 ml) was added and then stirred for 1
hours. 5-Heptyl-2,2'-bithiophene (0.5 g) dissolvedin
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WO 98I23269 PCT/US96/19118
dimethyl formamide (5 ml) was added and stirred at room
temperature for half an hour at room temperature, then the
temperature was raised to 60~C and was further stirred for
4 hours. The reaction solution was then extracted with
dichloromethane, neutralized with sodium acetate, washed
with water to neutrality, dried, concentrated and separated
by silica gel chromatography eluted with ethyl
acetate/n-hexane (1/15). Light yellowish oily product
(0.86 g, 86%) was obtained.
Spectral Data:
1H NMR (CDC13), 8 value
9.81 (s, iH, -CHO), 6.71-7.62 (m, 4H, protons of
thiophene)
2.79 (t, 2H, -CH2), 0.86 (t, 3H, CH3)
IR (neat) : c~ 1
2910 (CH), 1665 (C=O)
Mass spectrum,.m/e (relative intensity)
264 (M+, 44), 206 (100)
Pret~aration of 5-heptyl-2 2'-bithiophene
To a solution of 5-(heptan-1-one)-2,2'-bithiophene (2.0
g, 7.19 mmole) in dioxane (20m1), a mixture of hydrochloric
acid, dioxane and glacial acetic acid (15:20:15) was added.
Excess of freshly prepared zinc amalgam was then added and
stirred at room temperature for 2 hours. The solution
mixture was then extracted with ether, neutralized with 10%
sodium hydroxide aqueous solution, washed with water to
neutrality, dried, concentrated and separated by silica gel
chromatography eluted with n-hexane/ethyl acetate (1:20).
Oily product 0.91g (48%) was obtained.
Spectral Data:
1H NMR (CDClg), d value
6.65-7.24 (m,-5H, protons of thiophene), 1.67-123 (m,
2H, C_H2 ) ,
0.87 (t, 3H, CH_g) _
CA 02272275 1999-OS-18
WO 98I23269 - -- ' PCT/US96/19118
IR (neat): cm 1
2910 (CH)
Mass spectrum, (m/e) (relative intensity)
- 264 (M+, 24) , 179 (100)
Preuaration of 5-(heptan-1-one)-2.2'-bithiophene
To a solution of 2,2'-bithiophene (0.57 g, 3.43 mmole)
(Aldrich Chem. Co., Milwaukee, WI) in benzene (10 ml) was
added proper amount of phosphorous pentaoxide and was
to stirred at room temperature until the solution was
--. homogeneous. Heptanoic_acid (0.7 g, 5.35 mmole) was
dissolved in benzene (10 ml) and added slowly to the
reaction mixture. The solution was heated to 70~C for 2
hours. The solu~iQn was extracted with ethyl acetate,
15 washed with sodium bicarbonate aqueous solution several
times, then washed with water until neutrality. The
extract was dried-and concentrated. The residue was
separated by silica gel chromatography, eluted first with
hexane to recover starting material and then with
20 n-hexane/ethyl acetate (10:1). Light yellowish-crystalline
product 0.28g (30%-) was obtained, melting point 85~C.
Spectral Data:
1H NMR.(CDC13), a value ._-.
7.55-?.O1 (m, 5H, protons of thiophene), 1.75-1.20 (m,
25 8H, CH2) ,
0.85 (t, 3H, CH3)
IR (KBr): cm 1
1645 (C=O)
- Mass spectrum: m/e (relative intensity)
30 278 (M+, 20), 208 (40), 193 (51), 179 (100) --
- _ Example 2
Synthesis of l5-heptyl-5'-by oxvmethvl)-2.2'-bithiophene
- {NSC code # 647450) -
- 35 5-_Formyl-5'-heptyl-2,2'-bithiophene (0.4 g) was dissolved -
CA 02272275 1999-OS-18
WO 98/23269- PCT/US96/19118
36
in tetrahydrofuran (THF) (15 ml) and stirred at 0'C for 10
minutes. NaBH4 (0.1 g) was added at room temperature and
stirred for 2 hours. Then the solution was extracted with
ethyl acetate (100 ml). The extract was washed with water,
then dried and concentrated. The solid product was thus
obtained and further recrystallized from ethyl
acetate/n-hexane mixture. Light yellowish solid product
(0.21 g, 52%) was obtained and the melting point of the
product--was 59~C.
to Spectral Data: -~..~.._~-
1H NMR (CDC13), d value
.. 6.64-6.96 (m, 4H, protons of thiophene)
4.77-4.78 (m, 2H, OCH_2), 2.75 (t, 2H, CH2)
1.23-1.76 (m, 2H, CH2), 0.87 (t, 3H, CH_3) -
IR (KBr) : c~ 1
3250 (OH), 1070 (C-OH)
Mass spectrum, m/e (relative intensity)
294 (M+, 59), 209 (100)
Preparation of 5-formyl-2.2'-bithiophene
To dimethyl formamide (250 ml) was added phosphoryl
chloride (50.2 ml) with quick stirring. 2,2'-Bithiophene
. ( __ (83 g) in dimethyl formamide (200 ml) was then added and
stirred at- -10~C for 30 minutes. Then the temperature was
raised to 40~C and was further stirred far 20 hours. The
reaction mixture was then poured into crushed ice and
stirred for 30 minutes. Sodium hydroxide aqueous solution
(10%, 600 ml) was added and the solution was extracted with
chloroform. The organic layer was washed with water, dried
over anhydrous magnesium sulfate, concentrated and
separated by silica gel chromatography. Product 90 g was
obtained, melting point 56-57~C.
Spectral Data:
1H NMR (CDC13), 6 value
35_ 9.70 (s, iH), 7.60-7.47 (m, 1H), 7.30-7.10 (m, 3H),
- CA 02272275 1999-OS-18
WO 98/23269 PCT/US96/19118
37
'7~. 06-6.9.0 __0 1H) _
Example 3
Synthesis of 5-(1-hydroxypropyl)-2 2'-bithiophe~e
5-Formyl-2,2'-bithiophene (3 g) was dissolved in THF (50
ml) and ethyl-magnesium bromide Grignard reagent _(9.3 ml)
was added dropwisely under nitrogen gas atmosphere in ice
bath and stirred for 1 hour. The mixture was stirred for 1
hr. and then continuously stirred at room temperature. The
l0 reaction was monitored by thin layer chromatography. After
the reaction was completed, water (30 ml) and ethyl acetate
(200 ml) were added to the mixture and extracted and
purified by column chromatography, eluted with ethyl
acetate/n-hexane (1/9). Light yellowish product was thus
obtained.
Spectral Data: _.. _
IH NI~t (CDC13) , 8 value
7.16-6.63 (m, 5H, protons of thiophene), 4.78-4.73 (t,
1H, -CH(OH)CH2CH3)
1.92-1.79 (m, 2H, -CH2CH3), 0.98-0.90 (t, 3H, -C~-I3)
IR (NaCl) : cm-1
3400 (OH), 29,50 (CH) -
Example 4-....__
Svnthesis of 5-(3,4-dihydroxy-1-butenvl)-2 2'-bithiophene
5-(3-Oxo-but-1-en-4-al)-bithiophene (0.5 g) and NaBH4 (0.2
g) were dissolved in tetrahydrofuran (10 ml) and stirred at
room temperature for 2 hours. The reaction solution was
added with ethyl acetate (20o ml) and distilled water. The
30,extract was washed with water and dried over'anhydrous
magnesium sulfate: After concentration, the product was
separated by column chromatography, eluted with ethyl
acetate~n-hexane (3/7). Light yellowish crystals were thus
obtained and melting point thereof was 96-98~C.
CA 02272275 1999-OS-18
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38 -
Spectral Data:
1H NMR (400 MHz, CDC13), 8 value
3.60-3.74 (dd, 2H, -CH20H), 4.40 (m, iH, -CH(OH)),
5.98-6.06 (dd, 1H, C=CH-), 7.20-7.26 (d, 1H, -CH=C) :-
6.86-7.17 (m, 5H, protons of thiophene)
IR (KBr) : c~ 1
3326, 3076, 1462, 1423, 1122, 960, 790, 688
Mass spectrum (12~, m/e (relative intensity)
252 (M+, 91), 233(30), 221(100), 204(24), 176(15)
Pre~~aration of 5-(3-oxo-but-1-en-4.-al)-2 2'
-bithiophene -_
5-(4,4-Dimethoxy-3-oxo-1-butenyl)-2,2'-bithiophene was
dissolved in-methanol (40 ml). Hydrochloric acid (3 N, 20
ml) was_ added and refluxed for 4 hours. The reaction
mixture was extracted with ethyl acetate (400 ml). The
extract was washed with water and sodium bicarbonate
aqueous solution, dried, concentrated and separated by
silica gel chromatography. Dark orange oily product was
obtained.
Preparation of 5-(4,4-dimethoxy-3-hvdroxy-1-butenvl)-
2,2'-bithiophene
To a solution of 5-(4,4-dimethoxy-3-oxo-1-butenyl)-
2,2'-bithiophene (50 mg) in tetrahydrofuran (2.5 ml) was
added sodium borohydride (10 mg). The reaction mixture was
stirred at room temperature for 1 hour. Ethyl acetate and
water was added for partition. The organic layer was dried
over anhydrous magnesium sulfate, concentrated and
separated by silica gel chromatography. Light greenish
oily product was obtained.
Spectral Data:
13C ~ ( CDC13 ) , 8 value
140.91, 137.49, 136.25, 127.87, 127.00, 126.42, l25.04,
124.46,
CA 02272275 1999-OS-18
WO 98I23269 PCT/US96/19118
39
123.88 , 123.71, 71.83, 55.73, 55.11
IR ( neat ) : cta 1
3400 (O-H}, 2950 (C-H), 1070 (C-O)
Preparation of 5-(4.4-dimethoxv-3-oxo-1-butenyl)-
2 . 2' -l~ithiophene
To a solution of 5-formyl-2,2'-bithiophene (4.O g} in
ethanol (120 m1) was added dimethoxymethyl methyl ketone (3
ml). Potassium hydroxide aqueous solution (50%) was added
to dropwisely into the solution mixture at 16~C. Color
changed from yellowish brown into dark green, then dark
brown. lYellow solid was formed during the reaction. Water
was added. Solid residue was filtered and washed with
water, redissolved in acetone and separated by silica gel
chromatography eluted with ethyl acetate/n-hexane (1:9).
Yellowish crystals 1.88 g were obtained, melting point
74~C.
_ Spectral Data:
1H NMR (CDC13), 8 value
7.82 (d, lH, -CH=CH-), 7.27-7.03 (m, 5H, protons of
thiophene),
6.77 (d, 1H, -CH=CH-), 3.43 (s, 6H, (OCH3)2)
Mass-spectrum, m/e
294 (M+) , 219 (.M+-CH(OCH3) 2)
, - _ _
Example 5
Synthesis of 5-hydroxymethyl-5"-methyl-a-terthiophene
(NSC code # 647072)
5-Formyl-5"-methyl-a-terthiophene (0.2 g) was dissolved
in ethanol (50 ml). NaBH4 (0.1 g) was added at room
temperature and stirred for 30 minutes. The solution was
monitored by thin layer chromatography.._. After the reaction
was completed, 150 ml of ethyl acetate and 50 ml of water
were added therein. The ethyl acetate solution was washed
with water, dried and then purified by silica gel.column
CA 02272275 1999-OS-18
WO 98I23269 PCT/US96/19118
40 "
chromatographr~. Light yellowish crystals were thus
obtained and melting point thereof was 126-128~C. The
yield was almost quantitative.
Spectral Data:
1H NMR (CDC13), S value
7.09-6.72 (m, 6H, protons of thiophene)
4.87-4.85 (d, 2H, -CH20H), 2.54 (s, 3H, -CH3)
IR (KBr): cm 1
3300 (-OH), 2900(C-H), 1450 (conjugative C=C)
Mass spectrum, m/e (relative intensity)
292 (M+, 100), 275 (79), 258 (11)
Preparation of 5-formYl-5"-methyl-a-terthiophene
Phosphoryl chloride (1.0 ml) was added slowly to dimethyl
formamide (30 ml) at 0~C under nitrogen atmosphere. The
mixture was further stirred for 1 hour at 0~C._.. 5-Methyl-a-
terthiophene (0.5 g) in dimethyl formamide (5 ml) was added
dropwisely into the reaction mixture and stirred at room
temperature for 30 minutes. Temperature was raised to 60~C
and was further stirred for 2 hours. The reaction was
monitored by thin layer chromatography until no starting
material was left. The reaction mixture was then poured
iBto potassium carbonate aqueous solution with crushed ice
and extracted with ethyl acetate ~29~8 ml). The extract was
washed with water, dried, concentrated and separated by
silica gel chromatography, eluted with ethyl
acetate/n-hexane (1:4). The product was recrystallized
from ethanol to give orange crystals (90% yield), melting
point 158-159~C.
Spectral Data:
1H NMR (CDC13), v value
9.82 (s, 1H, CHO), 7.64-6.66 (m, 6H, protons of
thiophene),
2.47 (s, 3H, CH3) -
_35 IR (KBr): cm-1 -
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1670 (conjugated C=O), 1450 (conjugated C=C)
Mass spectrum: m/e (relative intensity)
290 (M+, I00) , 257 (9) , 217 (7)
Preparation of 5-methyl-a-terthioohene
5-Formyl-a-terthiophene (0:5 g) was dissolved in a
solution mixture of hydrochloric acid, dioxane and glacial
acetic acid (1:2:1) (50 ml). Zinc amalgam (5 g) was added
and stirred at room temperature for 3 hours. The reaction
was monitored with thin layer chromatography until no
starting material was detected, and then extracted with
- ethyl acetate (200 ml). The extract was washed with
potassium carbonate aqueous solution, dried,_concentrated
-- and separated by silica gel chromatography eluted with
n-hexane. The product was recrystallized from n-hexane to
give light yellowish crystals, melting point 93-94~C, yield
._ 90%. -
Spectral Data: _
1H NMR (CDC13), 8 value
7.19 (m, 7H, protons of thiophene), 2.46 (s, 3H, CH3)
IR (KBr): cm-1
2900 (O-H), 1430 (conjugated C=C)
Mass spectrum: m/e (relative intensity)
262 (M+, 100), 239 (17), 131 (12)
Preparation of 5-formyl-a-terthiophene and
5.5"-diformyl-a-terthiophene
(1) Phosphoryl chloride (1.0 ml) was added to dimethyl
formamide (15 ml)-and stirred for few minutes under
nitrogen atmosphere. a-Terthiophene (2.48 g) (Aldrich
Chem. Co., Milwaukee, WI) in dimethyl formamide was added
slowly and then heated to 70~C. The temperature was then
raised to 110~C and further stirred for 2.5 hours. Cooled
to room temperature and extracted with chloroform (10o ml). _ .
The extract was dried, concentrated and separated by silica .
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gel chromatography eluted with chloroform/n-hexane (1:4) to
give 5-formyl-a-terthiophene (1.94 g, 74.2%), melting point
141-142~C. Further elution with chloroform/n-hexane/ethyl
acetate (38:1:1) gave 5,5"-diformyl-a-terthiophene (0.13 g,
4.3%), melting point 219-220~C. a-Terthiophene (0.13 g)
was recovered.
Spectral Data:
5-formyl-a-terthiophene
1H NMR (CDC13), 8 value
9.86 (s, IH, CHO), 7.67(d, 1H; J=4Hz), 7.25 (d, 2H,
J=4Hz ) ,
7.21 (d, 2H, J=4Hz), 7.10 (d, 1H, J=4Hz), 7.02 (t, 1H,
J=4Hz)
Wmax '
Amax' 400rim
IR (KBr): cm-1
1649 (C=O), 2930 (C-H)
Mass spectrum: m/e
276 (M+)
5,5"-diformyl-a-terthiophene
1H NMR (CDC13), d value
9.86 (s, 2H, CHO), 7.67 (d, 2H, J=4Hz), 7.30 (s, 2H),
'7.27 (d, 2H, J=4Hz)
Wmax v
J~m~:_400nm -
IR (KBr): cm-1
1649 (C=O)
Mass spectrum: m/e
304 (M+)
(2) To a solution of 5-iodo-2-formylthiophene (1.0 g) in
acetonitrile (200 ml) was added 2,2'-bithiophene (0.7 g)
under nitrogen and irradiated with 100W.mercury lamp for 12
hours. The reaction was monitored with thin layer
chromatography until no further product was produced. The
solvent was removed and extracted with dichloromethane.
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The extract was dried, concentrated and separated by silica
gel chromatography eluted first with ethyl acetate/n-hexane
(1:9) to recover starting material, and then with ethyl
_ acetate/n-hexane (3:7) to obtained 5-formyl-a-terthiophene.
_
Example 6
Synthesis of 5-(pronan-1-ol)-a-terthiophene
(NSC code # 649663)
(1) 5-(Propan-1-one)-a-terthiophene (0.6 g) was dissolved
in ethanol (150 ml) and heated to dissolve completely.
NaBH4 (0.1-g) was added at room temperature and stirred for
3 hours. The solution was monitored by thin-layer _-
chromatography. After the reaction was completed, 50 ml of
water was added and the ethanol was removed under reduced
pressure. Yellowish green solid was thus obtained and
melted at 89-90~C. The yield was almost quantitative.
Spectral Data:
1H NMR (CDC13), d value -
7.20-6.84 (m, 7H, protons of thiophene)~
4.80-4.77 (t, 1H, -CH_(OH)-CH2CH3), 1.93-1.80 (m, 2H, _
-CH2CH3), 0.99-0.94 (t, 3H, -CH2CH3)
IR ( KBr ) : cm-1
3400G-OH), 2900(-CH), 1450 (conjugative C=C)
Mass spectrum, m/e (relative intensity)
304 (M+, 58), 277 (l00)
(2) 5-Formyl-a-terthiophene (3.5 g) was dissolved in
anhydrous THF (200 ml) under nitrogen stream. Ethyl
- Grignard reagent (7.9 ml) was dropped in slowly in ice bath
and stirred at 0~C--for 1 hour: The reaction mixture was
then heated to 70~C in oil bath for 4 hours. The solution
was monitored by thin layer chromatography. After the
reaction was completed, 50 ml of water and 300 ml of ethyl
acetate were added-.to extract the crude product, which was
then purified by column chromatography, eluted with ethyl
acetate/n-hexane (1/9). Yellow solid was then obtained and
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recrystallized to give granulate crystals. The melting
point of the product was 89-90~C and the yield was about
70%.
Preparation of 5-(propan-1-one)-a-terthiophene
To a solution of a-terthiaphene (5.0g) in benzene (250
ml) was added phosphorous pentaoxide (3.1g). Propanoic
acid (1.6 g) was then added slowly at room temperature.
The reaction mixture was heated to reflux and the reaction
was monitored with thin layer chromatography. A_little
more phosphorous pentaoxide was added to complete the
- reaction. Potassium carbonate aqueous solution was added
and extracted with ethyl acetate. The extract was dried,
concentrated and separated by silica gel chromatography,
eluted with ethyl acetate/n-hexane (1:19) to give yellow
solid product, which was recrystallized from ethyl
acetate/n-hexane (1:19) to give yellowish crystals (20%
yield), melting point 136-137C.
- Spectral Data:
1H NMR (CDC13), d value
7.58-7.00 (m, 7H, protons of thiophene), 2.89 (q, 2H,
-- _ CH2).
1.22 (t, 3H, CH3)
IR (KBr): c~ 1 -
2900 (C-H), 1650 (conjugated C=O), 1440 (conjugated C=C)
Mass spectrum: m/e (relative intensity)
304 (M+,73), 275 (100), 247 (21), 203 (59)
Euample 7
Svnthesis of 5-formyl-5"-~prgp-1~-enylZ-a-terthiophene
POC13 (2 ml) was added into dimethyl formamide (30 ml)
slowly under nitrogen gas atmosphere in ice bath and
stirred for 1 hour. Dimethyl formamide (20 ml) solution of
5-(prop-I-enyl)-terthiophene (0.8 g) was dropped in slowly. --
The mixture was stirred for half an hour at room
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temperature, then the temperature was raised to 60~C and
was further stirred for 2 hours. The solution was
monitored by thin layer chromatography. After the reaction
was completed, the reaction solution was poured into sodium
5 carbonate ice water solution. Then the solution was
extracted with 300 ml of ethyl acetate. After drying and
evaporation of the solvent residual solid was purified by
column chromatography, eluted with ethyl acetate/n-hexane
(1/9). _Orange solid product was obtained and the melting
10 point of the product was 146-148~C. The yield was about---~.----
. 7 0 % . _A._
-Spectral Data: -
1H NMR (CDC13), d value
9.50 (s, 1H, -CHO), 7.65-6.74 (m, 6H, protons of
15 thiophene)
6.47-6r43-(dd, 1H, -CH=CHCH3), 6a11-6.02 (m, 1H, __.
-CH=CHCH3)
1.86-1.84 (d, 3H, C~i3) ,
IR (KBr) : cia 1 _ _
20 2900(CH)., 1660(conjugative C=O), 1440 (conjugative C=C)
Mass spectrum, m/e (relative intensity)
3l6 (M+, 100), 149 (58)
Preparation of 5-(prop-1-envl)-a-terthiophene
25 5-(Propan-1-ol)-a-terthiophene (0.8 g) was dissolved in a
mixture of benzene/methanol _(1:1) (100 ml). Hydrochloric _
acid (2 N, 5 ml) was added at room temperature. Heated to
50~C for 1 hour and monitored with thin layer
chromatography until no starting material was left: Ethyl
30 acetate (250 ml) and water was added for partition. The
organic layer was dried, concentrated and separated by
silica gel chromatography eluted with n-hexane to give
desired product.
Spectral Data:
35 1H NMR (CDC13), 6 value
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7.19-6.72 (m, 7H, protons of thiophene), 6.45 (d, iH,
CH=CHCH3),
6.08-5.99 (m, iH, CH=CI_iCH3), 1.85 (d, 3H, CH=CHCH3)
IR (KBr): cm-1
2850 (C-H), 1430 (conjugated C=C)
Euample 8
Synthesis of 5-hydroxvmethyl-5~~-(proa-1-en~l1
-a-terthiophene
5-(Prop-1-enyl)-5~~-formyl-a-terthiophene (0.4 g) was
dissolved in ethanol (50 ml). NaBH4 (0.1 g) was added at
room temperature and stirred for 2 hours. The solution was
monitored by thin layer chromatography. After the reaction
was completed, water (5o ml) was added and the ethanol was
removed under reduced pressure. Then the solid precipitate
a was extracted with 200 ml of ethyl acetate. The
concentrated residual solid was purified by column
chromatography, eluted with ethyl acetate/n-hexane (3/7).
Yellowish solid product was obtained and melting point
thereof was 132-134~C. The yield was nearly 100%.
Spectral Data:
1H NMR (CDC13), 6 value
7.20-6.60 (m, 6H, protons of thiophene)
6.46-6.42 (dd, 1H, -CH=CH-CH3), 6.06-6.~1 (m, 1H,
-CH--CH_-CHg)
4.79-4.78 (d, 2H, -CH20H), 1.85-1.84 (d, 3H, -CH3)
IR (KBr): cm-1
- 3400 (OH), 2900 (CH), 1440 (conjugative C=C)
Mass spectrum, m/e (relative intensity)
3T8 (M+, 100), 301 (26), 261 (40)
Example 9
Synthesis of 5-ethyl-a-terthienylmethyl ether --
(1) 5-Formyl-a-terthiophene (0.3 g) was dissolved in
ethanol-(20 ml) by stirring at room temperature. To the
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solution, 0.04 g of NaBH4 was slowly added. After the
solution became clear in about 20 minutes, diluted
hydrochloric acid was slowly added until bubbling stopped.
The stirring was continued for about 12 hours, followed by
chloroform extraction and silica gel column chromatography
eluted with ethyl acetate/n-hexane (1/19). The product was
recrystalli2ed from chloroform/ethyl acetate mixture to
give light yellowish platelet crystals (melting point
76-77~C). The yield was about 41%.
(2) The yield could be increased to 85% or higher by
-.- substituting absolute ethanol for alcohol and concentrated
hydrochloric acid for diluted hydrochloric acid.
(3) 5-Formylterthiophene (2:9 g) was dissolved in
abso lute ethano l.,N.( 7_..5 ml ) . NaBH4 ( 0 . 5 g ) was added and
stirred for 10 minutes. The solution became clear-yellow.
Phosphorus oxychloride (2.5 ml) was added to an absolute _
ethanol and was dropped into aforementioned mixture and
stirred under the nitrogen atmosphere overnight. Then the
solution was extracted with ethyl acetate. The extract was
washed with water and dried over anhydrous magnesium
sulfate. The solvent was removed under reduced pressure.
The residual solid was purified by column chromatography.
Light yellowish crystals were obtained and the-in~lting
point thereof was 76-77~C. The yield was 85%.
Spectral Data:
1H NMR (CDC13), 8 value
7.20-6.87 (m, 7H, protons of thiophene), 4.62 (s, 2H,
-C_H20C2H5)
- 3.55 (q, 2H, -CH20C~i2CH3), 1.25 (t, 3H, -OCH2CH3)
3 0 IR ( KBr ) : ciri 1 _ _
3050 (aromatic CH), 2971, 2852 (saturated CH), 1091
_ ~C=O)
Mass spectrum, m/e (relative intensity)
- 306 (M+, 100)a 261 (M+-OC2H5, ~33)
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4 8 -
8aample 10
Synthesis of ethyl 5-(2,2~-bithienvl)-a-cyanoacrylate
5-Formyl-2,2~-bithiophene (3.9 g), cyano-ethyl-acetate
- (2.4 ml), pyridine (15 ml) and piperidine (2.4- ml) were
mixed together at 80-85~C for 2 hours. After cooling, the
mixture was acidified by diluted HC1 and then filtered.
Orange solid was thus obtained and recrystallized from
ethanol to give crystalline product (5.02 g). The melting
point of--the product was 131~C. The yield was 86%.
l0 Spectral Data: -- ,-~.---
IR (KBr) : cm-1
2218, 1687 (-CN)
1699, 1687 (C=O)
1630, 839, 796, 729 (thiophene) -
Mass spectrum, m/e (relative intensity)
289 (M+)
Example 11
Synthesis of methyl 4-(2.2~-bithiophen-5-yl)-2-oxo- _
but-3-enoate
4-(Bithiophen-5-yl)-2-oxo-but-3-enoic acid (0.62 g) was
dissolved in a mixture of methanol (25 ml) and benzene (15
ml). A trace amount of p-toulenesulfonic acid was added
- __ and heated for 6 hours. The solution was monitored by thin
layer chromatography. After the reaction was completed,
- 25 the solution was extracted with ethyl acetate and the
extract was washed with water and NaHC03 solution. The --
residual solid after evaporation was purified by column
chromatography, eluted with ethyl acetate/n-hexane (1/9).
Light yellowish crystals (0.63 g) were obtained and the
melting point of the product was 78~C. - The yield was 97%.
Spectral Data:
1H NMR (CDClg), d value
7.90-7.94 (d, 1H, -CH=CH-)
6.91-7.31 (m, 6H, -CH=CH-, protons of thiophene)
35. 3.89 (s, 3H, -COOCH3)
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IR (KBr) : cm-1 w _
1730, 1690 (C=O)
1650, 1600, 1580 (conjugative unsaturated bonding)
- Mass spectrum (75 ev), m/e (relative intensity)
278(M+, 16), 265(100), 219(93)
Euample i2
Svnthesis of 5.-a-terthienylmethy~idene malonic acid
5-Formyl-a-terthiophene (1.30 g), malonic acid (l.04 g),
pyridine (20 ml) and of piperidine (0.1 ml) were mixed
together at 50-60~C for 2 hours. Water (100 ml) was added
after reaction-was completed. The mixture was-acidified by
diluted HC1 and then filtered. Red solid thus obtained was
recrystallized-from ethanol to give 1.43 g of the product
and the=melting point thereof-was 202-203~C. The yield was
84%.
Euample 13
Svnthesis of 5-~(3-hydroxy-1-propynyl)-2,2~-bithiopphene
Hydroxy-1-propyne (48 mg) was dissolved in benzene (1
ml). 5-Iodo-2,2~-bithiophene (0.1 g) was added immediately
and stirred. A mixture of CuI G0.05 g),
benzyltriethylammonium chloride (0.05 g), catalyst Pd(PPh3)4
(0.1 g) and 3 ml of NaOH solution G2.5 N) were mixed
together at room temperature and stirred for 2 hours.
NH4C1 solution (4 ml) was then added. The reaction
solution was extracted with ethyl acetate. The extract was
washed with 10 ml of HC1 (10%) for 3 times, 50 ml of water
twice and dried over anhydrous magnesium sulfate. The
residual solid was purified by silica gel column
chromatography, eluted with ethyl acetate/n-hexane (3/1) to
yield 56 mg of the product (74%).
Spectral Data:
- 1H NMR 400 MHz (CDC13), d value
2.3,(b, 1H, -OH), 4.5 (s, 2H, -CH2) -
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6.9-7.3- (m, 5H, protons of thiophene)
13C-~ (CDC13) , d value
51.50, 78.89, 92.19, l21.11, 123.48, l28.06, 133.4,
136.70, 139.25
5 IR (neat): cm 1
3350 (OH}, 2250 (C=C)
Preparation of 5-iodo-2.2'-bithiophene and 5.5'-diiodo-
2.2'-biothiophene
10 To a solution of 2,2'-biothiophene (33.2g) in ethanol (50
-.- ml) was added iodine (20.3 g) in ethanol (200 ml). -When
iodine ( V ) oxide ( 6 . 7 g) in water ( 3 0 ml ) was -added
dropwisely. The reaction mixture was further stirred- at
room temperature_.~o,~.5 hours. Ethanol was removed and the
15 residue was dissolved in dichloromethane. The solution was
washed with sodium bicarbonate aqueous solution (10%, 150
ml x 2) then water-(200 ml x 2). Dried over anhydrous
magnesium sulfate and concentrated to give crude, products.
Distilled under vacuum (105-110~C/0.1 mm Hg) to give 5-
20 iodo-2,2'-bithiophene (36.8 g, 63%). The residue was then
separated by silica gel chromatography eluted with n-hexane
to give 5,5'-diiodo-2,2'-bithiophene (5.36 g, 6.4%),-
melting point 170~C. - ---
Spectral Data:
25 5-iodo-2,2'-biothiophene
1H NMR (CDC13), d value
7.20 (m, protons of thiophene)
Mass spectrum, m/e (relative intensity)
292 (M+, 100) , 165 (M+-I, 14)
30 5,5'-diiodo-2,2'-biothiophene - -
1H NMR (CDC13), d value
_ 7.05 (d), 6.7 (d)
Mass spectrum, m/e (relative intensity)
418 (M+, 100) , 291 (M+-I, 9)
- 35
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51
_ _ _ _ ~~ple 1~
- Synthesis of 5-(2,2'-bithiophen-5 mil) pron-2-
- yn-1-ol-acetate
5-(3-Hydroxy-1-propyne)-2,2'-bithiophene (0.8 g) was
dissolved in acetic anhydride (0.5 ml) and pyridine (1.5
ml) at room temperature and stirred for 2 hours. The
reaction solution was extracted with ethyl acetate. The
extract was washed with 10 ml of HCl (10%) for 3 times, 10
ml of NaOH solution for 3 times, 50 ml of water twice and
dried over anhydrous magnesium sulfate. The residual solid
after evaporation was purified by silica gel column
chromatography eluted with ethyl acetate/n-hexane (1/10)-r
Light brownish-yellow oily product (0.71 g, 73%) was
obtained.
Spectral Data:
1H NMR (CDClg), 8 value _ _
2.1l (s, 3H, -CH3), 4.90 (s, 2H, -CH2)
6.97-7.22 (m, 5H, protons of thiophene)
IR (neat): cm-1
2250 (C=C), 1745 (C=O), l225
Mass spectrum, m/e (relative intensity)
262 (M+, 100) , 202 (65) __
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Example 15
Synthesis of ethyl-5-a-terthien~rlprogiolate
Ethyl propiolate (78 mg) was dissolved in benzene (3 ml).
5-Iodo-a-terthiophene (0.2 g) was added immediately and
stirred. A mixture of CuI (0.05 g), benzyltriethylammonium ---
chloride (0.05 g), catalyst Pd(PPh3)4 (0.1 g) and 5 ml of
NaOH solution (2.5 N) were mixed together. The temperature
of the mixture was-t~.~.en raised to 40~C and stirred for 5
more hours. Then 5 ml of NH4C1 solution was added. The
reaction solution was extracted with ethyl acetate. The
- extract was washed with 10 ml of HC1--(10%) 3 times, 50 ml
of water twice and dried over anhydrous magnesium sulfate.
The residual solid after evaporation was purified by silica
gel column chromatography, eluted with ethyl
acetatefn-hexane (1/15). Light brown product (78 mg, 42%)
was then obtained and the melting point thereof was 93~C.
Spectral Data:
1H NMR (CDC13), d value - ..
1.35 (t, 3H, -CH3), 4.31 (q, 2H, -CH2)
6.90-7.41 (m, 5H, protons of thiophene)
IR (KBr): cm-1
2200 (C=C), 1705 (C=O)
Mass spectrum, m/e (relative intensity)
344 (M+, 17), 248(100)
Preparation of 5-iodo-a-terthiophene
a-Terthiophene (2.33 g) was dissolved in ethanol (70 ml).
Iodine (1.10 g) in ethanol (25 ml) was added and then iodic
acid (0.47 g) in water (1.0 ml) was added dropwisely and
the temperature was kept below 31~C. The reaction mixture
was further stirred for 4 hours. Precipitate from the
reaction was filtered and recrystallized to give product
(97.3% yield), melting point 146-148~C.
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53
Euample 16
Synthesis of 5-a-terthienylacrvlic acid
(NSC code # 637391)
. 5-Formyl-a-terthiophene (2?6 mg), malonic-acid (0.212 g),
pyridine (10 ml), and piperidine (1 ml) were reacted
together in water bath for 2 hours and refluxed for 30
minutes in oil bath and then cooled down to room
temperature. The reaction solution was then poured into
Water. The mixture was acidified by diluted HC1. After
standing at room temperature for 3 hours, the reddish-brown
solid was filtered, washed with water and recrystallized
from 95% of ethanol to give copper red needle-like crystals
and the_melting point thereof was 237-238~C._ The yield was
_ 78 . 6% . _.___
Spectral Data
UV: hmax: 395 nm
_ IR (KBr) : c~ 1
3200-2300 (OH)., 1672 (C=O),_ l616 (CH=CH)
1H NMR (d6 -DMSO): d value __
6.13 (1H,-CH=CH-COON, d, J=16), 7.70 (1H, -CH=CH-COOH,
d, J=16)
7.1-7.6 (m, 7H, protons of thiophene)
Mass spectrum, m/e (relative intensity)
318 (M+) . _
Example 17
Svn-thesis of methyl 4-(a-terthiophene-5-yl)-2-oxo-
but-3-enoate
4-(a-Terthiophene~5-yl)-2-oxo-but-3-enoic acid (0.2 g)
was dissolved in ethanol (20 ml):---A trace amount of
p-toluenesulfonic acid was added and refluxed overnight.
Then the solution was-extracted with ethyl acetate. The
extract was purified by column chromatography, eluted with
ethyl acetate/n-hexane (1/9). Dark reddish crystal product - -
was then obtained. -
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Spectral Data:
1H NMR (CDC13), d value
7.9-7.94 (d, 1H, -CH=CH-)
7.01-7.32 (m, 6H, -CH=CH-, protons of thiophene)
3.91 (5, 3h, -COOCH3)
Mass spectrum (75 ev): m/e (relative intensity)
360 (M+, 53), 301 (l00), 282 (23) _
Preparation of 4-(a-terthiophene-5-yl~~-2-oxo-but-3-enoic
to acid --
To a solution of 5-for~riyl-a-terthiophene (0.5 g) in
ethanol (200 ml) was added a solution of sodium pyruvate
(0.3 g) in water (3 ml) at room temperature. Then 50%
sodium hydroxide solution (1 ml) was added and stirred at
room temperature for over night. After acidification with
10% hydrochloric acid, the resultant was extracted with
ethyl acetate (200 ml x 2). The extract was condensed
under reduced pressure and the darkish solid obtained was
used for methyl esterification.
Example is
Synthesis of 5-formyl-a-tetrathiophene
tNSC code # 645273)
Dimethyl formamide (30 ml) was stirred and cooled in ice
bath for 10 minutes. POC13 (0.2 ml) was added into the
solution at 0'C and stirred for i hour. Dimethyl formamide
(20 ml) solution of a-tetrathiophene (0.5 g) was dropped in
slowly. The mixture was stirred for 1 hour at 0'C, then
the temperature was raised to room temperature, and further
stirred for 8 hours at 70'C. The cold reaction solution
was poured into NaOH ice water solution, and extracted with
dichloromethane. The extract was washed with 50 ml of
water for 3 times, dried over anhydrous magnesium sulfate
and concentrated. The residual solid was further purified
by silica gel column chromatography, eluted with__
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n-hexane/tetrahydrofuran (1/1). Dark yellow solid (0.22 g,
41%) product was obtained after recrystallization from
n-hexane and tetrahydrofuran. The melting point of the
product was 215'C.
5 Spectral Data:
1H NMR (CDC13), d value
7.00-7.66 (m, 9H, protons of thiophene)
9.84 (s,: 1H, -CHO)
IR ( KBr ) : cm-1
10 1665 (C=O)
Mass spectrum, m/e (relative intensity)
358 (M+, 100) _-
Preparation of a-tetrathiophene
15 5-Iodo-2,2'-bithiophene (10 g, 34.2 mmole) and copper-
powder (2.8 g, 44.0 mmole) was mixed together. Dimethyl
formamide (20 ml) was added and heated to reflux for 6
hours. The reaction mixture was extracted with
tetrahydrofuran and decanted to separate the unreacted
20 copper powder. The solvent was removed and the residue was
separated by silica gel chromatography eluted first with
n-hexane to recover starting material and then with
n-hexane/tetrahydrofuran (2:1) to give product. Light
yellowish crystals were obtained after recrystallization
25 (34.8% yield), melting point 208~C.
Euample 19
w Synthesis of 5-hydroxymethyl-a-tetrathioohene
(NSC code # 645274)
30 5-Formyl-a-tetrathiophene (180 mg) was dissolved in
dimethyl formamide (10 ml) solution . NaBH4 (0.1 g)was then
added. The mixture was stirred for 2 hours at room
temperature. Then.the solution was extracted with
dichloromethane and washed with 10 ml of water twice and
35 dried over anhydrous magnesium sulfate. After evaporation
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the residual solid was purified by silica gel column
chromatography and further recrystallized from THF and
n-hexane. Light orange solid (150 mg, 82.4%) was obtained
and the melting point of the product was 216~C.
- 5 Spectral Data:
IR (KBr): cm 1 _ _
3300 (OH)
Mass spectrum, m/e (relative intensity)
360 (M+, 100)
344 (21)
- Example 20
S~rnthesis of 5- ( 4-hvdroxy-1-butynyll -2 , 2' -bi-thiophene
(1) Hydroxy-1-butyne (0a23 g) was dissolved in benzene (1
ml). 5-Iodo-2,2'-bithiophene (0.3 g) was added immediately
and stirred. CuI (0.05 g), benzyltriethylammonium chloride
(0.05 g), Pd(PPh3)4 (0.1 g) and NaOH solution (2.5 N) (4 ml}
were mixed together at room temperature and stirred for 2
hours. NH4C1 solution (5 ml) was then added. The reaction
solution-was extracted with ethyl acetate. The extract was
washed with l0 ml of HC1 (10%) for 3 times, 50 ml of water
twice and dried over anhydrous magnesium sulfate. The
residual solid after evaporation was purified by silica~gel
column chromatography, eluted with ethyl acetate/n-hexane
(1/3). Light yellowish product (0.21 g, 75%) was obtained
and the melting point thereof was 67~C.
(2) In a brown three-necked round bottom flask equipped
with condenser, thermometer and nitrogen inlet was added
5-iodo-2,2'-bithioghene (17.07 g) and pyridine (100 ml).
3-Butyn-1-of cuperous salt was qui-ckly added and refluxed
under nitrogen atmosphere for 3.5 hours. Pyridine was then
distilled and the reaction mixture was extracted with
dichloromethane (100 ml x 2). The extract was washed with
water (150 ml x 2}, 10% sodium bicarbonate aqueous solution --
(100 ml x 2) and again water (150 ml x 2) to remove water
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soluble compounds. The organic layer was dried over
anhydrous magnesium sulfate, concentrated and separated by
silica gel chromatography eluted with n-hexane to recover
- starting material (3.56 g) and then with n-hexane/ethyl
acetate (6:1) to give light yellowish crystalline product
(8.49 g, 62%).
Spectral Data:
1H NMR (CDC13), d value
7.20--6.75 (m, 5H, protons of thiophene), 3.7 (t, 2H,
CH20H) , __ ~_
. . 2.7 (t, 2H, CH2CH20H) v,..
IR:- _ cm-1
3300 (br, O-H), 1032 (C-O)
Mass spectrum: m/e (relative intensity)
234 (M+, 17), 203 (M+-CH20H,_ 10), 84 (100)
Example 21
Synthesis of 5-(4-acetoxy-1-butynyl)-2.2'-bithioghene _
4-(Hydroxy-1-butynyl)-2,2'-bithiophene (1.23 g) was
dissolved in pyridine (6 ml). Acetic anhydride (1 ml) was
added at room temperature, stirred for few minutes and
further kept overnight. Water (20 ml) was added oily
product was_-_then extracted with 30 ml of ethyl acetate.
The extract was washed with 10 ml of HC1 (1N) for 3 times,
25_. 10 ml of water once and 10 ml of- diluted KHC03 solution
once. Ethyl acetate was then removed under reduced -
pressure. The residual solid was purified by silica gel
column chromatography, eluted with ethyl acetate/n-hexane
(1/19). Light yellowish oily product (1.3 g) was obtained
after removal of the solvent. -
Spectral Data:
1H NMR: d value
7.3-6.8 (m, 5H,-protons of thiophene), 4.16 (t, 2H,
-CH2-CH2-O-)
_ 35._ 2.7 (t,--2H, -CH2-CHz-O-), 2.0 (t, 3H, -OCOCH3) _
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IR: cm 1
3035 (aromatic C-H)
2960, 2900 (adipose C-H)
1737, 1232, 1038 -(ester)
Mass spectrum, m/e (relative intensity)
276 (M+, 46), 2l6 (M+-AcOH, 100)
Euample 22
Synthesis of 5-(4-isovaleryloxy-1-butynyl)-2,2'-
to bithiophene
5-(4-Hydroxy-1-butynyl)-2,2'-bithiophene (1 g) was
dissolved in pyridine (10 ml). Isovaleroyl chloride (l ml)
was added at room temperature, stirred for few minutes and
kept overnight. The reaction mixture was treated as in
example 21. Light yellowish oily product (1.06 g) was then
obtained.
Spectral Data:
1H NMR: 8 value
6.95-7.1 (m, 5H, protons of thiophene), 4.2 (t, 2H,
J=7Hz, -CH2-CH2-O-) _.
2.72 (t, 2H, J=7Hz, -CH2-CH2-O-), 2.18 (br.s. 2H,
-O-CO-CH2-.)
2.2-1.9 (m,'1H, -CH2-CH(CH3)2), 0.96 (1, 6H, J=6Hz,
CH(CH3)2) __
IR: cm'1
3100_, 3070 (aromatic C-H), 2960-2870 (aliphatic C-H)
1730, 1250, 1150 (ester)
1460, 1380, l360 (-CH(CHg)2)
834, 795, 692-(2,2'-bithiophene)
Mass spectrum, m/e (relative intensity)
318 (M+, 15), 216 (M+-(CH3)2CHCH2COOH, 100)
- EBample 23
Synthesis of 5-(4-benzoxy-1-butynvl)-2,2'- bithiouhene
5-(4-Hydroxy-1-butynyl)-2,2'-bithiophene (0.8 g) was _
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dissolved in pyridine (8 ml). Benzoyl chloride (i ml) was
added at room temperature, stirred for few minutes and kept
overnight. The reaction mixture was treated as in example
_ 21. Light yellowish crystals were then obtained and the
_ 5 melting--point thereof was 61~C.
Spectral Data:
IH NMFt: 6 value _
6.86-8.28 (m, 10H, aromatic H) -
4.42 (t, 2H, -CH2-CH2-O-), 2.85 (t, 2H, -CH2-CH2=O-)
IR: cm 1
3090, 3040 (aromatic C-H)
1695, 1268, l109 (aromatic ester)
Mass spectrum, m/e (relative intensity)
388 (M+, 16) , _21f....(M+-C6HSCOOH, 100)
- 15
Example 24
Synthesis of 5-(4-palmityloxy-1-butynyl)-2.2'- bithiophene
5-(4-Hydroxy-1-butynyl)-2,2'-bithiophene (0.6 g') was
dissolved in pyridine (10 ml). Palmitoyl chloride (1 ml)
was added at room temperature, stirred for few mixiutes and
kept overnight. The reaction mixture was treated as in
example 21, and 1.2 g of light yellowish crystals were then
obtained and the melting point thereof was 68-69-~-C.
Spectral Data:
1H NMR: 8 value
7.26-6.84 (m, 5H, protons of thiophene)
4.25 (t, 2H, -CHZ-C~i2-O-), 2.67 (t, 2H, -CH2-CH2-O-)
2.28 (t, 2H, -OCOCH2C14H29),_ 1.22 broad (m, 29H, _
_ - -OCOCHZC14H29)
IR: Cai 1
3030 (aromatic C-H), 2950, 2840 (aliphatic C-H)
- 1730, i170 (ester)
Mass spectrum, m/e (relative intensity) -
- 472 (M+, 25), 216 (M+-C15H31COOH, 100) -
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Example 25
Svnthesis of 5-(3-hydroxy-4-pyranyl~l-1-but~nyl-2.2'-
bithiophene
4-(Tetrahydropyranyloxy)-3-hydroxy-butyne (6.7 g) was
5 dissolved in benzene (20 ml). 5-Iodo-2.2'-bithiophene
(5.75 g) was added immediately and stirred. A mixture of
CuI (0.15 g) and benzyltriethylammonium chloride (0.14 g,
0.63 mmole~ was added. Pd(PPh3)4 (0.46 g, 0.398 mmole) was
then added. NaOH solution (30 ml, 2.5 N) was added slowly
10 into the mixture at room temperature in water bath for 2---~----
hours. NH4C1 solution (lo ml) was added. The reaction
solution was extracted with ethyl acetate. The extract was
washed with 10 ml of HC1 (10%) for 3 times, 50 ml of water
for 3 times and dried over anhydrous magnesium sulfate.
15 The residual solid after evaporation was purified by silica
gel column chromatography, eluted with ethyl
acetate/n-hexane (1/3). Reddish oily product (6.25 g, 95%)
was then obtained.
Spectral Data:
20 1H NMR (CDC13), d value
1.4-1..9 (m, 6H, 3CH3), 3.3-4.0 (m, 4H, 2CH2)
4.5-4.7 (m, 1H, CH), 6.9-7.2 (m, 5H, protons of
thiophene)
_.. IR ( neat ) : c~ 1
_ 25 3400 (OH), 2950(C-H), 2250(C=-C)
Mass spectrum, m/e (relative intensity)
334(80, M+), 304(41), 234(49), 190(51), 86(100)
30 Preparation of 3-hydroxy-4-(tetrahydro-2-
pyranyloxy)-butvne
Acetylene gas was bubbled into tetrahydrofuran (30 ml)
for 30 minutes. Ethyl magnesium bromide (25 ml) was added.
2-(Tetrahydro-2-pyranyloxy)-acetaldehyde (5.0 g, 0.035
35 mole) in tetrahydrofuran (25 ml) was then added. The
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61
reaction mixture was warmed to room temperature and stirred
overnight. Saturated ammonium chloride aqueous solution
(30 ml) was added to quench the unreacted acetylide and
extracted with ethyl acetate. The extract was dried over
anhydrous magnesium sulfate, concentrated and separated by -
silica gel chromatography eluted with n-hexane/ethyl
acetate (5:2) to give colorless oil (4.5 g, 76%).
Spectral Data:
1H Nl~t ( CDC13 ) , 6 value
4.6-4.4 (m, 2H, 2CH), 3.8-3.6 (m, 2H, CH2), 3.5-3.4 (m,
2H, CH2 ) ,
2.4-{s, 1H, C=CH), 1.9-1.4 (m, 4H, 2CH2) _ -
IR (neat): cm-1
3600-3000-{O-H), 2150 (C=C)
Mass s~aectrum: m/e (relativeintensity)
l69 (M+-1), 115 (13), 97 (19), $5 (l00), 56 (48)
Preparation of 2-(tetrahydro-2-ovranyloxy~-- r
- acetaldehyde
2-(Tetrahydro-2-pyranyloxy)-propene (20 g, 0.14 mole) was
dissolved in dichloro-methane (250 ml). Ozone was bubbled
into the solution at -78~C for about 3 hours until the
solution turned blue. Warmed to room temperature and zinc
powder (20 g) was added. Acetic acid (20 ml) and water (3
ml) were added slowly with cooling in ice bath. The
reaction mixture was further stirred for 2 hours at room
temperature and then extracted with dichloromethane. The
extract was washed with sodium bicarbonate aqueous solution
(10 ml x 2), dried over anhydrous magnesium sulfate,
3o concentrated. The crude product was distilled under vacuum
- (62-64~C/_2 mmHg) gave colorless oil (13.72 g, 68%).
- 2-(Tetra-hydro-2-pyranyloxy)-propene was directly prepared
from allyl alcohol and dihydropyran in the presence of a
small amount of p-toluenesulfonic acid.
Spectral Data:
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a
1H Nl~t (CDC13) , 6 value
9.7 (s, 1H, CHO), 4.6 (t, 1H, O-CH-O), 1.9-1.3 (m, 6H,
CH2)
13C ~ (CDC13) , 6 value
200._78, l00.89, 74.29, 63.89, 31.39, 26.35, 20.35
IR ( neat ) : cai 1
2950 (C-H), 1735 (C=O)
Example 26
l0 Synthesis of 5-f3-acetoxy-4-tetrahvdropyranyloxyy-1-
_.. butynyl-2.2'-bithiophene _
3-Acetoxy-4-tetrahydropyranyloxy-1-butyne (4-.73) was
dissolved in benzene (15 ml). 5-Iodo-2,2'-bithiophene
(5.15 g) was added immediately and stirred. A mixture of
CuI (0.l3 g), benzyltriethylammonium chloride (0.12 g) and
catalyst Pd(PPh3)4 (0.4 g) was then added. NaOH solution
(20 ml, 2.5 N) was-added slowly into the mixture at room
temperature for 2 hours. Saturated NH,~C1 solution (8 ml)
was added. The reaction mixture was extracted with ethyl
acetate. The extract was washed with 10 ml of HCl (10%)
twice,50 ml of water twice and dried over anhydrous
magnesium sulfate. The residual solid after evaporation
was purified by silica gel column chromatograph~,eluted
with ethyl acetate/n-hexane (1/3). Light orange product
(4.1 g, 67%) was obtained.
Spectral Data:
1H NMR (CDC13), d value
1.4-1.9 (m, 6H, 3CH2), 2.1 (s, 3H, CH3)
3.7-4.0 (m, 4H, 2CH2), 4.7-4.8 (s, 1H, OCHO) -
5.?-5.9 (m, 1H, CH), 7.0-7.3 (m, 5H, protons of
thiophene) __
IR (neat): cm 1
2950 (C-H), 2225 (C=C), 1750 (C=O)
_35 Mass_spectrum, m/e (relative intensity)
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376 (M+,_6~ ~ 333(21), 275(30), 234(100)
- Preparation of 3-acetoxv~,-(tetrahydro-2-
. _ v gYranvl, oxy~ -butyne
3-Hydroxy-4-(tetrahydro-2-pyranyloxy)-butyne -(8.0 g,
0.047 mole) was mixed with acetic anhydride (10 g, 0.098
mole) and pyridine'(8 g, o.098 mole). The solution was
stirred at room temperature for 1 hour and then extracted
with ethyl acetate. The extract was sequentially washed
with 10% hydrochloric acid (10 ml x 5), saturated sodium
bicarbonate aqueous solution (10 ml x 5) and water (50 ml x
3). The extract was dried over anhydrous magnesium -
sulfate, concentrated and separated by silica gel
chromatography, eluted with n-hexane/ethyl acetate (5:1) to
give colorless oil (9.0 g, 90%).
Example Z7
Synthesis of 5-(3-acetoxv-4-hvdroxy-1-butynyl)-2.2'-
bithiophene
(NSC code # 645277)
5-(3-Acetoxy-4-tetrahydropyranyloxy-1-butynyl)-2,2'-
bithiophene (2.8 g) was dissolved in methanol.~40 ml).
HZS04 solution (3 ml, 2N) was added slowly in ice bath and
stirred at room temperature for 2 ho~rrs. The reaction
solution was extracted with ethyl acetate. The extract was
thus washed with 10 ml of NaHC03 three time, 50 ml of water
twice and dried over anhydrous magnesium sulfate. After
removal of solvent residual solid was purified by silica
gel column chromatography, eluted with ethyl
acetate/n-hexane (1/3). Light brown product~(1.83 g, 84%)
was obtained.
Spectral Data:
1H NMR ~CDC13), 8 value
- 2.15 (s, 3H, -CH3), 3.9-0 (d, 2H, -CH2)
~5 5.71 (t, 1H, CH), 7.0-7.3 (m, 5H, protons of thiophene)
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IR (neat): cm 1
3450 (OH), 2250 (C=C), 1730 (C=O)
Ezampls 28
Svnthesis of 5-f3.4-diacetoxv-1-butyr~yl)~-2,2'- bithionhene --
5-(3-Acetoxy-4-hydroxy)-butynyl-2,2'-bithiophene (1.83 g)
was dissolved in pyridine (1.5 g) and acetic anhydride
(1.27 g).at room- temperature for 2 hours. The reaction
solution was extracted with ethyl acetate. The extract was
washed with 10 ml of HC1 (10%) for 3 times, 10 ml of KHC03
solution for three times and 50 ml of._water twice. The
solution was then dried over anhydrous magnesium sulfate.
After removal of solvent residual solid was purified by
silica gel column chromatography, eluted with ethyl
acetate~n-hexane (1/5). Greenish yellow oily product (1.91
g, 91%) was then obtained.
Spectral Data:
1H NMR (CDC13}, 8 value _- e,,
2.07 (s, 3H, -CH3), 2.I2 (s, 3H, -CH3)
4.25-4.45 (m, 2H, CH2), 5.80-5.87 (m, 1H, CH)
7.0-7.3 (m, 5H, protons of thiophene)
IR ( neat ) : cap 1
2250 (C=C), 1755 (C=O), 1735 (C=O)
Mass spectrum, m/e (relative intensity)
334 (M+, 75), 274(l00), 232(55)
Euample 29
Synthesis of 5.5'-dihydroxymethyl-2-2'-bithiophene
(NSC code # 647452)
(1) 5-Hydroxymethyl-5'-formyl-2,2'-bithiophene (0.2 g)
was dissolved in ethanol (50 ml). NaBH4 (0.1 g) was added
at room temperature and stirred for 1 hour. The reaction
was monitored by thin layer chromatography. After the
reaction was completed, H20 (50 ml) was added and the
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ethanol was removed under reduced pressure. Light
yellowish solid product was obtained. The yield was almost
quantitative and the melting point of the product was
l58-160~C.
5 Spectral Data:
1H NMR (d6-acetone)
7.03-6.87 (m, 4H, protons of thiophene)._
4.73 (s,-4H, -CH20H)
IR (KBr) : cm-1
10 3500-3300 (OH), 3050 (aromatic-CH)
2909, 2850 (aliphatic CH)
Mass spectrum, m/e (relative intensity) .
226 (M+, 100.), 209 (M+-OH, 73)
(2) 5-Hydroxymethyl-5'-formyl-2,2'-bithiophene (0.6 g)
15 was dissolved in tetra-hydrofuran (30-ml),-NaBH4 (0.16 g)
was added, and the solution was stirred for 2 hours at room
temperature. The solvent was removed under reduced
pressure. White solid obtained was washed with water and
dried under reduced pressure. The yield was quantitative.
20 The melting point of the product was 155-156~C.
(3) 5-Hydroxymethyl-5'-formyl-2,2'-bithiophene (0.5 g)
was reduced in ethanol (75 ml) with NaBH4 (0.3 g). The
mixture was stirred for 3 hours atroom temperature. The
solution was concentrated and n-hexane was added to obtain
25 white_pr~duct. The-product was washed with water, dried
under reduced pressure and the-yield was quantitative. The
melting point of the product was 155-156~C.
(4) 2-Hydroxymethyl-5-iodothiophene was refluxed with Cu
powder in dimethyl formamide. The Ullmann condensation
30 also produced 5,5'-dihydroxy-methyl-2,2'- bithiophene in
low yield.
(5) The Ullmann condensation of 2-acetoxymethyl-5-
iodothiophene gave 5,5'-diacetoxymethyl-2,2'-bithiophene.
The 5,5'-dihydroxymethyl-2,2'-bithiophene was obtained by
35 alkaline hydrolysis of diacetoxy compound and,purified by _
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6 6 --
column chromatography. The yield was about 20%.
- E$ample 30
Synthesis of 5.5'-diacetoxvmethvl-2-2'-bithiophene
5,5'-Dihydroxymethyl-2,2'-bithiophene (0.23g), pyridine
(1.2 ml) and acetic anhydride (0.3 ml) were mixed, stirred
and kept overnight. Then the mixture was extracted with
ethylacetate. The pyridine and acetic acid were removed by
washing with weak base, weak acid and water, respectively.
Silica gel powder was added into the ethyl acetate solution
and the solvent was removed under reduced pressure. Coated
silica gel powder was added to the silic gel column and
chromatographed. The eluant was ethyl acetate/n-hexane
(7/3). The white platelet crystal thus obtained was
further recrystallized with ethyl acetate/n-hexane mixture.
The melting point of the product was 60~C.
Spectral Data:
IR: cm 1
l725 (C=O)
Mass spectrum, m/e (relative intensity)
310 (M+, 37)
251 (M+-CH3C02, 100) __
192 (M+-2CH3C02, 34)
Example 31
Synthesis of 5-k~ydroxymethyl-5'-formyl-2-2'-bithiophene
(NSC code # 647073)
Phosphorus oxychloride ("POC13") (1 ml) was added into
dimethyl formamide (20 m1) slowly under nitrogen gas
atmosphere in ice bath and stirred for 1 hour. The
dimethyl formamide solution- (5 ml) of 5-hydroxymethyl-
2,2'-bithiophene (0.5 g) was dropped in slowly. The
mixture was stirred for half an hour at room temperatures--
then the temperature was raised to 50~C and was further
~ 5 stirred for 3 hours. The reaction solution was poured into
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potassium carbonate ice water solution. Then the solution
was extracted with 100 ml of ethyl acetate. The extract
was washed with water and dried over anhydrous magnesium
- sulfate. The solvent was removed under reduced pressure,
and the residual.solid was purified by column _
chromatography, eluted with ethyl acetate/n-hexane (3/7).
Light yellowish product was recrystallized from ethyl
acetate/n-hexane mixture. The melting point of the product
was 123-124~C. The yield was 85%.
Spectral Data: _
1H NMR 400 MHz (CD~13), d value
9.91 (s, 1H, -CHO), 7.73-7.02 (m, 4H, protons of
thiophene) -
4.91-4.90 (d, ZH, -CH20H)
IR ( KBr ) : c~ 1
3300 (OH), 1640 (C=O)
Mass spectrum, m/e (relative intensity)
224 (M+, 100), _207 (M+ -OH, 57), l95 (M+ -OH- -CHO, 22)
Preparation of 5-hvdroxymethyl-2,2~-bithiophene
To a solution of 5-formyl-2,2'-bithiophene (20 g) in
methanol (50 ml) was added sodium borohydride. The
ruction mixture was stirred at room temperature and
monitored with thin layer chromatography until the reaction
completed. After removal of methanol, water was added to
dissolve inorganic salts and extracted with
dichloromethane. The extract was washed with brine, dried
over anhydrous magnesium sulfate, concentrated to give
_ product (20.1 g, 99%~), melting point 52-53~C.
Spectral Data: -
-- 1H NMIt ( DMSO-D6 ) ( d va lue
' 7.4-6.6 (m, 5H), 5.51---(t, 2H), 4.5 (d, 2H)
Wmax v
- 320-300nm
IR: c~a 1 _ -
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3250 (O-H)
Mass spectrum: m/e
196 (M+)
68
Example 32
Synthesis of 5-acetoxymethyl-5'-formyl-2-2'-bithiophene
(NSC code # 647453) _
- 5-Hydroxymethyl-5'-formyl-2,2'-bithiophene (0.2 g) and
pyridine (1 ml) were mixed together. Acetic anhydride (1
l0 ml) was added slowly into the miX-ture with stirring. Ethyl
acetate (200 ml) and water (50 ml) were added 2 hours
later-. _ The ethyl acetate layer was washed with weak base,
weak acid and water. The product was concentrated and
purified by column chromatography, eluted with ethyl
acetate/n-hexane (1/9). Light yellowish crystals were
obtained. The melting point of-the crystal was 89-91~C.
The yield was 95%.
Spectral Data:
1H NMR 400 MHz (CDC13), d value
9.83 (s, 1H, -CHO), 7.64-7.01 (m, 4H, protons of
thiophene)
5.20 (s, 2H, -CH20Ac), 2.08 (s, 3H, -COCI_i3)
IR _ ( KBr ) : cm 1
1740, 1660 (C=O)
2 5 _ __ - -
Example 33
Synthesis of 5-hvdroxymethyl-5"-formyl-a-terthiophene
(NSC code # 647455)
POC13 (1 ml) was added to dimethyl formamide (30 ml)
slowly under nitrogen stream in ice bath. The solution was
stirred for 0.5 hour and then dimethyl formamide solution
(20 ml) of 5-hydroxymethyl-a-terthiophene (0.3 g) was
dropped in slowly. The mixture was stirred for an hour at
room temperature and then the temperature was raised to
60~C and stirred for 2 more hours. The reaction-.solution
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69
was poured into ice aqueous potassium carbonate solution.
The solution was extracted with 300 m1 of ethyl acetate and
the extract was washed with water. The solvent was removed
. J- under reduced pressure and the residual solid--was purified
by column chromatography, eluted with ethyl
acetate/n-hexane (3/7). Orange crystals were obtained and
the melting point of the pxoduct was 176-177~C. The yield
was 80%.
Spectral Data:
1H NMR ( CDC13 ) , d value __ ..
9.86 (s, 1H, -CI-~O), 7.65-6.91 (m, .bH, protons of
~thiophene)
4.80 (s, 2H, -C~-I20H)
IR (KBr): cm 1
3400 (OH), 1660 (C=O)
Mass spectrum, m/e (relative intensity)
306 (M+, 100), 289 (M+,-OH, 56)
Preparation of 5-hydrox<rmethyl-a-terthiophene
To a solution of 5-formyl-a-bithiophene (0.5 g) in
tetrahydrofuran (20 ml) was added sodium borohydride (0.034
g). The reaction mixture was stirred at room temperature
_. _._ ., for 2 hours and monitored with thin layer chromatography
until the reaction completed. Water (50 ml) was added
slowly and extracted with chloroform. The extract was
dried over anhydrous magnesium sulfate, concentrated to
give yellowish powder, melting point 151-152~C, yield 97%.
spectral Data:
1H NMR (DMSO-d6), d value
7.51 (d, 1H, J=4Hz), 7.31 (dd, 1H, J1=1, 4Hz), 7.24 (d,
1H, J=4H2), 7.20 (d, 1H, J=4H2), 7.15 (d, 1H, J=4Hz),
7.09 (dd, 1H, J=3.5, 4HZ), 6.91 (d, 1H, J=4HZ), 5.52 (d,
1H, J=6H2), 4.60 (d, 2H, J=6H2)
Wmax
35Jlm~: 355nm
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IR (KBr) : cm 1
3300 (O-H), 3061 (=C-H), 2950 (-C-H), 1060 (C-O)
Mass spectrum: m/e
278 (M+)
5 ~ -
Ezample 34
Synthesis of 5,5"-dihydroxymeth~rl-a-terthiophene
(NSC code a 646270)
5,5"-Diformyl-a-terthiophene (1 g) was added into
10 tetrahydrofuran (150 ml). The temperature was raised to
50~C until the solid was completely dissolved, then NaBH4
(0.25 g) was added and stirred for 3 hours at-50~C. The_
solvent was removed under reduced pressure. Ethyl acetate
and water were added to dissolve the residual solid. The
15 ethyl acetate layer was washed with water and dried over
anhydrous magnesium sulfate. The ethyl acetate layer was
filtered and concentrated to obtain light yellowish
crystals (0.95 g) and the melting point of the product was
182-183~C.
20 Spectral Data:
1H NMR 400 MHz (CDC13), 6 value
7.04-6.89 (m, 6H, protons of thiophene)
4.79 (d, 4H, CH20H), 1.51 (br. s., OH)
Mass spectrum, m/e (relative intensity)
25 308 (M+, 58), 306 (M+-2H, 100)
Example 35
- Synthesis of 5-hydroxymethyh-5"-(1-hydroxypropyl)-
a-terthiophene
30 (1) POC13 (0.5 ml) was added to dimethyl formamide (3~
ml) slowly under nitrogen stream in ice bath condition.
The solution was stirred for 1 hour and dimethyl formamide
solution (200 ml)_~f 5-hydroxymethyl-a-terthiophene -(0.8 g)
was dropped in slowly. Then heated to 70~C with oil bath
35 for 3 hours. The reaction solution was thus placed into 50
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ml of aqueous K2C03 solution at 0~C. The solution was
extracted with 500 ml of ethyl acetate. The residual solid
was purified by column chromatography, eluted with ethyl
acetate/n-hexane (3/7). After removal of solvent, the
residue-was dissolved in 50m1 of tetrahydrofuran under
nitrogen stream and dropped in 1 ml of Ethyl Grignard
reagent and stirred at room temperature for 3 hours. The
solution was monitored by thin layer chromatography. After
the reaction was completed, then the solution was extracted
with 300 ml of ethyl acetate and 50 ml of water. After
__. removal of solvent, the residual solid was purified by-
column chromatography, eluted with ethyl acetate/n-hexane
(3/7). Orange powder was thus-obtained and the melting
point of the product4was 131~C.
(2) 5-Hydroxymethyl-5"-formyl-a-terthiophene (0.5 g) was
dissolved into anhydrous tetrahydrofuran (50 ml). A
slightly excessive amount of ethyl magnesium bromide (2.0
M) was added to the solution under nitrogen atmosphere.
The solution was stirred for 3 hours at room temperature.
Aqueous ammonium chloride solution was added to hydrolyze
the above reaction solution to obtain-the crude product.
The crude product was collected and purified by column
chromatography, elute-with ethyl acetate/n-hexa-ire (3/7).
The eluate was concentrated to obtain orange powder (0.3
g). The melting point was 131-132~C.
Spectral Data:
1H NI~t (CDC13) , 6 value
7.03-6.85 (m, 6H, protons of thiophene) _
4.79-4.78 (m, 3H, -CH20H and -CHC2H5)
1.91-1.77 (m, 2H, -CHZCH3), 0.99-0.9 5 (t, 3H, -CH3) --
IR (KBr): c~ 1
_ 3400 (OH), 2900 (saturated CH)
Exstnple 36 _
_35_ Synthesis of 5-succinoylmeth~yl-2,2~-bithiophene
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5-Hydroxymethyl-2,2'-bithiophene (2.5 g), pyridine (20
ml) and succinic anhydride (1.2 g) were mixed together and
stirred at 40~C. Thin layer chromatography was applied to
monitor the reaction. After the reaction was--completed,
ethyl acetate and diluted hydrochloric acid were added.
The reaction solution was concentrated and then mixed with
n-hexane for crystallization. White crystals (2.43 g) were
then obtained and melting point thereof was 112~C.
Spectral. Data:
1H NI~t ( CDC13 ) : d value --~---~
7.20-6.90 (m, 5H, protons of thioghene), 5.23 (s, 2H,
_ ~ -CI_i20- )
2.69-2.61 (m, 4H, -COCH2CH2C0-), 2.40 (br, OH)
IR (KBr) : c~ri 1 -
3200-2500 (OH)
1720, 1690 (C=O)
Example 37
Synthesis of 5.5~-disuccinovlmethyl-2,2~-bithiophene
5,5~-Dihydroxymethyl-2,2~-bithiophene (0.5 g), pyridine
(10 ml) and succinic anhydride (2 g) were mixed together.
The mixture was stirred at 40~C. Thin layer chromatography
- - ' was applied to monitor the reaction. After the reaction
was completed, ethyl acetate was added to extract the
-, 25 product. The ethyl acetate layer was washed with diluted
hydrochloric acid and water in order to remove pyridine -
completely. The product was filtered through silica gel
and recrystallized from ethyl acetate/n-hexane. White
crystals (0.45 g) were obtained. The melting point of the
crystal was 137~C. _
Spectral Data:
1H Nl~t (CDC13) , 8 value
7.00-6.90 (m, 4H, protons of thiophene), 5.28-5.23 (m,
4H, -0I20-)
_ 35_ 4.78-4.75 (m, 4H, -CH20-), 2.69-2.64 (m, 8H,
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-CO-CH_2 CI_i2-CO- ) -
IR (KBr): cm 1
3600-2500 (OH)
- 17I8, 1688 (C=O)
Example 38
Synthesis of 5-succinovlmethvl-5'-formvl-2.2'-bithioohene
5-Hydroxymethyl-~--formyl-2,2'-bithiophene (0.63 g),
pyridine (10 ml) and succinyl anhydride (0.12 g) were mixed
together. The mixture was stirred at 40~C. Thin layer
_ chromatography was applied to monitor--the reaction. After
the reaction was completed, diluted hydrochlar-ic-acid and
ethyl acetate were added. The ethyl acetate solution was
washed with water to remove pyridine completely. Then the
ethyl aZ:etate layer was dehydrated with anhydrous magnesium
sulfate and filtered through silica gel layer. After
removal of the solvent, the product was recrystallized from
ethyl acetate/n-hexane to give a light yellowish crystals
(0.6 g). The melting point was 127~C.
Spectral Data:
1H Nl~t ( CDC13 ) , S value
9.84 (s, 1H, -CH_O), 7.65-7.02 (m, 4H, protons of ._
thiophene)
5.25 (s, 2H, -CH20-), 2.72-2.64 (m, 4H, -COCIi2C~2C0- )
2.40 (br, OH)
IR ( KBr ) : cap 1
3200-2500 (OH)
1730, 1705, 1650 (C=O)
Example 39
Synthesis of 5-formyl-2.2':5',3"-terthiophene
(NSC code # 660643)
In a two-necked round bottomed flask was added
5-dimethoxymethyl-5'-tributyl-stannyl-2,2'-bithiophene (5.6
g), bis(triphenylphosphine)palladium (II) chloride (0.32
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g), 3-bromo-thiophene (1.5 g) (Aldrich Chem. Co.,
Milwaukee, WI) and tetrahydrofuran (20 ml). The reaction
mixture was refluxed for 16 hours. To the reaction mixture
was then added hydrochloric acid (1 N, 3 ml) and was
further-refluxed for 3 hours. Saturated sodium bicarbonate
aqueous solution was added to neutrality and extracted with
ethyl acetate. The extract was washed with brine, dried
over anhydrous magnesium sulfate and concentrated. The
residue was separated by silica gel chromatography, eluted
with n-hexane/ethyl acetate (9:1 -~ 1:1 gradient) to give
-- brownish solid product (0.7 g, 40%), melting point -
155-157~C.
Spectral Data:
1H NMR ( CDC13 ) , d ..ua.lue
9.86 (s, 1H), 7.67 (d, 1H, J=3.8H2), 7.44-7.23 (m, 5H),
7.15 (d, 1H, J=3.8Hz)
IR (CH2C12) : cm-1 -
1695 (C=O) --
Mass spectrum, m/e (relative intensity)
276 (M+, 100), 270 (12), 247 (8), 203 (24), Y2'7 (6)
Preparation of 5-dimethoxvmethyl-5'-tributvlstannvl-
2.2'-bithiophene -
To a solution of 5-dimethoxymethyl-2,2'-bithiophene (1.0
g) in tetrahydrofuran (20 ml) with ice bath cooling was
added n-butyllithium (1.6 M in hexane, 3.0 ml). The ice
bath was removed and the reaction mixture was stirred at
room temperature for 1 hour. Then cooled with ice bath
again and tributyltin chloride (1.14 ml) was added._ The
reaction mixture was further stirred at room temperature
for 4 hours. Solvent was removed and the residue was
- quickly filtered through aluminum oxide eluted with
n-hexane to give desired product (1.6 g).
Spectral Data:
-35 1H NMR (CDC13), a value -
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7.28-7.26--(~_1H), 7.06-7.04 (m, 2H), 6.97-6.94 (m,
1H), 5.61 (s, lH)-
- 3.38 (s, 6H), l.62-0.87 (m, 27H) .
5 Preparation of 5-dimethoxvmethyl-2.2'-bithioBhene
Trimethyl orthoformate (7.5 ml) and montmarillonite K-10
(5.0 g) (Aldrich Chem. Co., Milwaukee, WI) were mixed and
stirred at room temperature for l0 minutes. A solution of
_ 5-formyl-2,2'-bithiophene (5.0 g) (Aldrich Chem. Co.,
10 Milwaukee, WI) in n-hexane (10 ml) was added and was
stirred at room temperature. The reaction was monitored
with thin layer chromatography until the reaction was
completed. Montmorillonite K-10 was filtered. The
filtrate was added to a saturated sodium bicarbonate
15 aqueous solution and extracted with ethyl acetate (15 ml x
3). The extract was washed with brine, dried over
anhydrous magnesium sulfate and concentrated to give light
yellowish oily product (6.4 g).
Spectral Data:
20 1H NMR (CDC13), d value
7.19-7.13 (m, 2H), 7.05 (d, iH, J=4.OHz), 7.00-6.94 (m,
2H) , _ _
5.59 (d, 1H, J=0.5H2), 3..37 (s, 6H)
25 Example 40
Synthesis of 5-hydroxymethyl-2.2':5',3"-terthiophene
(NSC code # 660644)
To a solution of 5-formyl-2,2':5',3"-terthiophene (0.5 g)
in methanol (15 ml) was added excess amount of sodium
30 borohydride. The reaction mixture was stirred at room
temperature for 40 minutes. Methanol was removed and water
was added. The solution was extracted with ethyl.acetate;
washed with brine, dried over anhydrous magnesium sulfate-
- and concentrated to give-brownish solid product (0.5 g),
35 melting point 158-160~C.
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Spectral Data: ' -
1H NMR (CDC13), 8 value
7.40-7.27 (m, 3H), 7.11-7.03 (m, 2H), 6.92 (d, 1H,
J=3.49Hz), 4.81 (s, 2H), ,,
1.62 (brs, OH_)
IR (CHC13) : cm 1
3650 (O-H)
Mass spectrum, m/~e-relative intensity)
278 (M+, 100), 261 (55), 245 (9),_ 216 (8), 203 (15), 127
(5)
Example 41
Synthesis of 4-formvl-5-hydroxymethyl-2.2':5',2"-
terthiophene -- -
(NSC code # 663562)
In a two-necked round bottomed flask was added
5-tributylstannyl-2,2'-bithiophene (2.04 g),
bis(triphenylphosphine)palladium (II) chloride (0=.19 g),
3-formyl-2-hydroxymethyl-5-iodo-thiophene (1.0 g) and
tetrahydrofuran (20 ml). The reaction mixture was refluxed
for 16 hours. Extracted with ethyl acetate, washed with
brine, dried over anhydrous magnesium sulfate and
concentrated. The residue was separated by silica gel
chromatography, eluted with n-hexane/ethyl acetate (9:1 -.
1:1 gradient) to give brownish solid product (0.43 g, 38%),
melting point 108-110~C. -
Spectral Data:
1H NMR (CDC13), d value
10.01 (s, 1H), 7.56 (s, 1H), 7.45 (dd, 1H, J=1.0,
5.2Hz),
7.32 (dd,.lH, J=1.06, 4.7H2), 7.25 (q, 2H), 7.09 (dd,
iH, J=5.04, 5.2Hz), 5:21-5.15 (m, 3H)
IR ( CH2C12 ) : c~a 1
w - 3630 (O-H), 1690 (C=O)
Mass spectrum, m/e (relative intensity)
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306 (M+, 100), 277 (22), 249 (29), 216 (21), 171 (13),
127 (13) , 108 (10)
_ Preparatior~ of 3-form~rl-2-hvdrox~nethyl-5-
iodothiophene
To a solution of 3-dimethoxymethyl-2-formylthiophene (8.8
g) in methanol was added excess amount of sodium
borohydride and stirred at room temperature for 2 hours.
Methanol was removed and residue was dissolved in ethyl
acetate. The solution was washed with water, dried over
anhydrous magnesium sulfate and concentrated. The crude
intermediate was then dissolved in methanol (40 ml).
Iodine (4.5 g) was added and then iodic acid (2.2 g) in
water (10 ml)was added dropwisely. The solution mixture
was stirred at room temperature for 4 hours. Methanol was
removed and sodium thiosulfate (10%) was added to quench
the unreacted iodine. Extracted with ethyl acetate, washed
- with brine, dried over anhydrous magnesium sulfate,
concentrated and separated by silica gel chromatography to
give yellowish oily product (10.4 g, 70%).
Spectral Data:
1H NMR (CDC13), 6 value
9.87 (s, 1H), 7.61 (s, 1H), 4.98 (s, 2H), 3.49 (s, 1H)
Preoar-ation of 3=dimethoxymethyl-2-formylthiophene
To a solution of 3-dimethoxymethylthiophene (14 g) in
tetrahydrofuran (8o ml) was slowly added n-butyllithium
(1.6 M in n-hexane, 47.8 ml) at -10~C with stirring. The
_ reaction mixture was further stirred at same temperature
for l hour. N,N-Dimethyl-formamide (11.9 g) in
tetrahydrofuran (40 ml) was then added. The reaction
mixture was warmed to room temperature-and stirred for
overnight. Ice water was added and extracted with ethyl
acetate. The extract was dried over anhydrous magnesium
sulfate and concentrated to give brownish oily product (13 _ --
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g, 80%). _.
Spectral Data:
1H NMR (CDC13), d value
10.25 (d, iH, J=1:16H2), 7.65 (dd, 1H, J=1.05, 4.03H2),
7.24 (d, 1H, J=5.12H2), 5.82 (s, 1H), 3.38 (s, 6H)
Euample 42
Synthesis of 4,5-dihydroxymethyl-2.2':5'.2"-terthiophene
(NSC code # 663561)
To a solution of 4-formyl-5-hydroxymethyl-2,2':5',2"-
terthiophene (0.1 g) in methanol (10 ml)i was added excess
amount of sodium borohydride. The reaction mixture was
stirred at room temperature for 40 minutes. Methanol was
removed and water was added. The solution was extracted
with ethyl acetate, washed with brine, dried over anhydrous
magnesium sulfate and concentrated to give brownish solid
product (0.1 g); melting point 99-101~C.
Spectral Data:
1H NMR (CDC13), d value
7.42 (dd, 1H, J=6.29H2), 7.28 (dd, 1H, J=1.13, 3.61H2),
7.20-7.13 (m, 3H),
7.07 (dd, lH, J=4.92, 5.29H2), 4.77 (d, 2H, J=5.64H2),
4.57 (.d, 2H, J=5.70H2)
4.28 (t, OH), 3.68 (t, OH)
IR (CHC13) : Cm 1
3630 (O-H)
Mass spectrum, m/e (relative intensity)
- 308 (M+, 100), 291 (34), 262 (24), 261 (23), 217 (18),
2I6 (12), 171 (10),
127 (11)
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Example 43
- ( Synthesis o~ 5-formvl-4"-hvdroxvmethyl-
2.2':5'.2"-terthiophene
- (NSC code # 658879)
In a two-necked round bottomed flask was added -
5-dimethoxymethyl-5'-tributyl-stannyl-2,2'-bithiophene
(2.12 g, 4.0 mmole), bis(triphenylphosphine)-palladium (II)
chloride (140 mg, 0.2 mmole), 2-iodo-4-
hydroxymethylthiophene (0.96 g, 4~0 mmole) [prepared from
l0 sodium borohydride reduction of 2-iodo-4-formyl-thiophene
(R. Guilard, P. E~urnari, and M. Person, Bulletin De La
Societe Chimique De France, 11, 4121, 1967) in
tetrahydr~furan, 95% yield] and tetrahydrofuran (25 ml).
The reaction mixture was refluxed for 16 hours. To the
reaction mixture was then added hydrochloric acid (1 N, 3
ml) and was further refluxed for 3 hours. Saturated sodium
bicarbonate aqueous solution was added to neutralize the
solution and extracted with ethyl acetate. The extract was
washed with brine, dried over anhydrous magnesium sulfate
and concentrated. The residue was separated by silica gel
chromatography, eluted with n-hexane/ethyl acetate (2:1) to
give yellowish solid product (420 mg, 34%), melting point
133-135~C.
Spectral Data:
1H NMR (CDC13), d value
9.86 (s, 1H), 7.67 (d, 1H, J=8Hz), 7.26-7.22 (m, 3H),
7.16-7.11 (m, 2H),
- 4.68 (s, 2H)
IR (CH2C12) : cm-1
3620 (O-H), 1680 (C=O)
- Mass spectrum, m/e (relative intensity)
306 (M+, l00) , 233 (9) ,- 69 (8) , 28 (15)
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EZa:ple 44
Synthesis of 5.4"-dihvdroxvmethvl-2,2':5'.2"-terthiophene
(NSC code # 658878) __
To a solution of 5-formyl-4"-hydroxymethyl-2,2':5',2"-
5 terthiophene (140 mg) in methanol (10 ml) was added sodium
borohydride (0.1 g). The reaction mixture was stirred at
room temperature for 40 minutes. Methanol-was removed and
water was added. The solution was extracted with ethyl
acetate, washed with brine, dried over anhydrous magnesium
10 sulfate and concentrated to give light yellowish solid
product (140 mg), melting point 111-113~C. _
Spectral Data:
1H NMR (CDC13), d value
7.18-7.02 (m, 5H), 6.96-6.89 (m, 1H), 5.30 (s, 1H), 4.81
15 (brs, 2H), 4.67 (s, 3H)
IR ( CHC13 ) : cm 1
3650 (O-H)
Mass spectrum, m/e (relative intensity)
308 (M+, l00) , 292 (14) , 29l (60) , 275 (10)
Facample 45
Synthesis of 5-formyl-3"-hydroxymethyl-2.2':5'.2"-
terthiophene
(NSC code # 658876)
In a two-necked round bottomed flask was added
5-dimethoxymethyl-5'-tributyl-stannyl-2,2'-bithiophene
(0.77 g), bis(triphenylphosphine)palladium (II) chloride
(0.06 g), 3-hydroxymethyl-2-iodothiophene (0.38 g) and
- tetrahydrofuran (20 ml). The reaction mixture was refluxed
for 16 hours. To the reaction mixture was then added
hydrochloric acid (1 N, 10 ml) and was further refluxed for
3 hours. Saturated sodium bicarbonate-aqueous solution was
added to neutrality and extracted with ethyl acetate. The
extract was washed with brine, dried over anhydrous
magnesium sulfate and concentrated. The residue--was _
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separated by silica gel chromatography, eluted with
n-hexane/ethyl acetate (9:1 -~ 1:1 gradient) to give
brownish solid product (0.24 g, 50%), melting point
' 85-86~C. _ _
Spectral Data:
. 1H NMZt (CDC13) , S value
9.86 (s, 1H), 7.68 (dd, 1H, J=1.46, 3.96Hz), 7.32-7.24
(m, 3H) ,~
7.18-7.16 (m, 2H), 4.80 (s, 2H), 1.64 (bs, OH)
IR (CH2C12) : cm 1
3630 (O-H), 1680 (C=O)
Mass spectrum, m/e (relative intensity)
306 (M+, 100), 289 (23), 273 (13), 227 (11), 203
(11), 171 (18),-121 (15)
Prepa~n of 3-hydroxymethyl-2-iodothiophene
3-Hydroxymethylthiophene (2.3 g, 20 mmole) (Aldrich Chem.
Co., Milwaukee, WI) and iodine (2.39 g, 9.4 mmole) were
dissolved in ethanol (15 ml). Iodic acid (l.09 g, 6.2
mmole) in water (2 ml) was added dropwisely at 0~C and was
further stirred for 1 hour at the same temperature. The
reaction was. monitored with thin layer chromatography until
the reaction was completed. Sodium thiosulfate aqueous
solution was added and extracted with dichloromethane. T he
extract was washed with brine, dried over anhydrous
magnesiu;n sulfate, concentrated and separated by silica gel
chromatography, eluted with n-hexane/ethyl acetate (3:1) to
give light yellowish oily product (3.9 g, 81%).
Spectral Data:
1H NMR (CDC13), d value
7.43 (d, 1H, J=5.2Hz), 6.96 (d, 1H, J=5.2Hz), 4.54 (s, __
' 2H)
Example 46
~,ynthesis of 5.3"-dihydroxyx~ethy~-2.2':5',2"-terthiophene .
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(NSC code # 658875)
To a solution of 5-formyl-3"-hydroxymethyl-2,2:5',2"-
terthiophene (60 mg) in tetrahydrofuran (5 ml) was added
sodium borohydride (15 mg). The reaction mixture was_
stirred at room temperature for 40 minutes.
Tetrahydrofuran was removed and water was added. The
solution was extracted with ethyl acetate, washed with
brine, dried over anhydrous magnesium sulfate and
concentrated to give yellowish solid product (60 mg),
melting point 93-95~C.
Spectral Data:
- 1H NMR (CDC13), S value
7.38 (d, 1H, J=5.3Hz), 7.21-7.13 (m, 3H),_6.92 (d, 1H,
_ J=2. 65Hz) , -._.
4.76 (d, 2H, J=6.OHz), 4.71 (d, 2H, J=5.53Hz), 4.59 (t,
1H, J=5.5Hz, OH),
4.35 (t, 1H, J=5.43Hz, OH)
IR (CHC13) : cm 1
3640 (O-H)
w~- - 20 Mass spectrum, m/e (relative intensity)
308 (M+, 100) , 291 (44) , 277 (13) , 227 (11) , 171 (13)-,
l21 (13)
Example 47
The Preparation of Tartaric Acid Salt of
Aminomethylterthiophene:
To an ethanolic solution (20 mL) containing
aminomethylterthiophene (300 mg, 1.08 mmol) was dropwisely
added, with stirring, aqueous L-tartaric acid (2.160 mL, 05
M solution prepared from L-tartaric acid (600 mg, 4.0 mmol)
in 8 mL of H20). Ethanol was removed under vacuum and the
residue was freeze-dried to give the salt (460 mg).
The Preparation of (2:1) Hydroxypropyl-~-cyclodeutrin _
Compleu of the Tartaric Acid Salt of -
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Aminomethylterthiophene:
To a solution containing hydroxypropyl-p-cyclodextrin
(3.60-g) in H20 (50 mL) was added tartaric acid salt of
' ~ aminomethylterthiophene (460 mg, 1.08 mmol) in one portion
and s.~nicated for 10 minutes. The heterogeneous mixture
was heated to near boiling with stirring until a clear-
_ solution was resulted. The solution was cooled to room
temperature, gravity filtered, and freeze-dried to give
(2:1) hydroxypropyl-p-cyclodextrin complex of the tartaric
acid salt of aminomethylterthiophene.
The Quantitative Calculation of the Amount of
Aminomethylterthiophene in (2:1) Hydrouypropyl-/9-
cyclodextrin Complex of the Tartaric Acid Salt of
Aminomethylterthiophene: -
To 25.21 mg of the (2:1) cyclodextrin complex was added -
1.10 mg (0.0079 mmol) of nitrophenol and was dissolved in
D20 ( 1 mL) for 1H Nl~. The ration of
aminomethylterthiophene to nitrophenol calculated from the
1H NIA integration_ was 89%. - _
0.0079 mmol (nitrophenol) x 0.89 = 0.00704 mmol
(aminomethylterthiophene)
- w 0.00704 mm~l x 277g/mol (m. wt. of aminomethylterthiophene)
- 1.95 mg
- 25 Since 1.95-;ng of aminomethylterthiophene is in 25.21 mg
of the cyclodextrin complex,~l.0 mg of
aminomethylterthiophene is in 12.93 mg of the cyclodextrin
complex.
The solubility of cyclodextrin complex in H20 was tested
by dropwisely adding H2o to 30 mg of the cyclodextrin _
complex, with stirring. The result showed that the 30 mg
of the cyclodextrin complex, containing 2.32 mg of
aminomethylterthiophene, can be dissolved in 30 ~tL of H20
- to give clear syrup. The result suggests that 1 mL of H2o
- 35- can dissolve up to_77 mg of aminomethylterthiophene (1000 -
CA 02272275 1999-OS-18
WO 98l23269 PCT/C1S96/19118
84 _
mg of the cyclodextrin complex) to give clear syrup.
Ezample 48
Other polythiophene-compounds prepared in accordance with
this invention are those represented by the following
structural formulas.
_ CA
02272275
1999-OS-18
WO 98/23269 -
PCT/L1S96/19118
85
I ( ~~~H~H
S I
I
S
_ ( I I CH=CHCOOCH3
I
S S
1 o I I I CHaCHHCOOEt
I
S S
I I I I ~CH=CHCOOH
I
__ S S
5.
I I I cooH
~
~Hs~\
S S ~pH
__
.
_.
2 o I I
I (
CHO
S S
I I I
( I I
S S ~HCH2CHCH:OH
$
~
_
25 OH
_
CHCH3
S S S
i
OH
3 ~ I I I I _
I
t
_ S~ $ ~~H3
$
ii
_ f I I I I
I CHrS-C-NH2
S S S
CA 02272275 1999-OS-18
WO 98/23269 - - PCT/US96/19118
86
S S
- OH
CHZOH
I I
-.. HOH2C S~S _ . .
CEiO
HOH2C ~ ~ I I
S S
oHC ~- ~ ~ I ~o _
S S
CHO
OHC I I ~ ~O
- S S S
-
HOH2C ~ I ( ~ ~CHZOH
S S S
CA 02272275 1999-OS-18
WO 98/23269 PCT/US96/19118
- I I I . .-.
I cH2pH
_ S _
S
I i I I . cH off
I ~ =
'
s s
s
I i I I cH=pcocx,
I I
$ S
$
I I I I ~
I I
S S
S
I I ~cHzpTs
I
S S _.. _
C f
I
~-cocH3
s s
CH~OC ! ~ CpCH3
I
I
S
S
1 I cN
I
I
I
I
S S
S
_ _
I I II
S S
S
I
I.
COOH
S
S _
_. .. I , _
I
I
!
I
I I
I
I
cH~Bri
__
- CA 02272275 1999-OS-18
WO 98l23269 ~ PCT/US96119118-
88
~CHZI~'H
S S
-~
.-
I I
I I
S S
CHaN-CH=CFiCHZOH
OH
I I
l p I I
~=hTHCHZCHC-Fi=OH
S -S
_
H
I I
~c=~=~
i I
S S
II S
O
I I
I I
~_~~-~'
S S
~~
O
I I
I I
~_~~
I I
_
S S
~~
S
~ .
i I
I ~
cH=cH-c
H
S S
O
i ~
~ I
CH,
CH=GH
C
CH
-
~-
y
S S
O OH
I i ~3 _
I
S S
CH=CH-C-CH
O OCH~
I I
i cH,
CH=CH-CH--C-CH3
- S
S
OH OH
CA 02272275 1999-05-18
WO 98/Z3269 - PCT/US96/19118
89
S I I S I CMG-CH-CH=THP
_ 5
I I I S ( ~_~~I
S OH OH
i o _ , _ ~, I I I I ~-o.~-~=oH _
s s ~
- I ( ~c~c-cH-CH=
S S p
I I I ~C--CH--CH=TaP
S. S bH
I ( I ~c~.C oN
_ S S COiH
__ ( _ _) _ I I ~~C CN
S S 'CN
I I I ~ C00&
CH=C'~
S S 'COOEc
I I I I oHsG'Cr
S S COOEc --.
_ I ( I I I I CO:H _.
CH=C~
S S S 'CO=H - _ .
_ CA 02272275 1999-OS-18
WO 98/23269 - - PCT/US96i19118
I ~ ~C$c-CH-CH=OH
s s
OH
i S I I S ~ ~C-CHCH=OCOCITHss
OCOCI~H3s
1~ I I i l I ~ I ~ _ _ ._
Q-iZOH
S S S S
I ~ ~ I I I ~ r.~0 _ .
S S S S
I S I i S ~ ~-~~soH
OCOCI~H3s
i
2 0 __
I ~--~c-cHCH=ococl7H,s
S S ~ _ .__
H
_. ( I ~ ( ~C ~CH20COC1~Hss
~coc H
11 73
I I I C=C--COiEc .. _
S
l I ~C=C-CH-CHs
- S S OH
~I I I ~~C-Cri:OH . _
~~g .
-CA 02272275 1999-OS-18
WO 98I23269 - PCT/L1S96/19118
91
I I I I I I CH=CHCOOH
S -
S S
I I I I I I
S S ~=N~
O
_
_
S
' I I I I I I
S S oH~~H
S -
I I I I
S I GCCOZCH~CH3
S S
-
HOH2C I I I I
S I
I
CHIOH
_.
_
S S
_ I I I _ ( off=OOH
_
S S -
_ ,
--
2~
l I
I I CH=NOH
-
S S
_ I I I I CH=CH--CH-CH=OAc
S S
I CH=CH~F-i--CFi20H
I I
I
S OH
S
I I I
cx=CH-CH-cH~oH
s S OAc
- I I I r o~,c
-
-cH=CH-cH-cH
S S =
~
pH
_
.
CA 02272275 1999-OS-18
WO 98l23269 PCT/US96/19118
92
OHC ~ $ ~ ! I I ~ ; CHCH2CH3
$ $ b
H
Hoxlc I S ! I S ( I S ! cxcHIcH3
bx
- - T. _ $ S $
I - I I I ~ CHCH2CH3
$ $ $
H
l ~ ~ ~ C(CH~sp
S S II
N-OH
2 0 ~ ( I I ~ pOCH CH OH
$ $ 2 2 2
_ _- p p
CHZOCCHz-C"FiI-C-0N~(C2Hs)3
S S
I I I I I ( CH OCCH CH flH
$ $ $ 2 2 2
3 0 OHC I I ~ I CH OOCH CH OH
$ $ 2 2 2
O O O _ O _
HOCHZCHZCCOHZC ! S ( I $ I CH20CCHZCH2COH
CA 02272275 1999-05-18
WO 98/Z3269 - PCT/US96/19118
93
- ~ 'S~--~sj~-'S~--CHIOCH=CH=OCH3
H3C I I ~ I I ( ~O
S S S
H3C ( S I I S I I S I CHIOH
HOH=C ~ I I ~--CHO
S S
_. CH3(CFi~~6 I I I I CHO __
S S
CH3(CH=)6 I S ~ I S ( ~ZOH
I I i I ~Cx~~sCx,
Sr' 'S'
_ O
__ __..HOCHz-~ I ~ CH=OH
S S
OHC ~ ( __ . ( I CH=OCOCH3
so ~ I I 1 I I
S S S ~~ - .
HOH2C I . I ( I I ~--C~iO
S S S
CA 02272275 1999-OS-18
WO 98I23269 PCT/US96/19118
94
CHZOH
HOHIC I ( I - I I I
S 'S ~~S ~~CH:OH
20H
I I I I I ~ _
to S S ~S~~CHZOH
._ _ ___ CH20H -
HOHZC I I I I I I CH20H _
S S S
CHO
__
_ S~S~S~~O
CHO
S S S . CH=OH
20H
so ~ ~ ' I ~ I
OHC S S S ~O
~C--cH20A~
CA 02272275 1999-OS-18
WO 98/23269 - - PCT/US96/19118
Example 49
Additional National Cancer Institute data demonstrating
polythiophene growth inhibition of human cancer cell lines
-- is represented in the following tables. The compound must
5 exhibit a LoglO GI50 value of <-4.00 to be considered active
against the tested cell line.
N8C: 659567-F/0-l/34
10 _
s s s II _
.
Panel/Cel1 Line Loglo GI50 LogIOTGI Loglo
LC50
Leukemia _
CCRF-CEM -4.84 -4.47 - -4.09
HL~O(TB) -5.03 -4.47 >-4.00
2 0 K=562 -S.70 >-0.00 >-4.00
MOLT-4 -4.58 >-4.00 >-4.00
_ .- . RPMI-8226 -4.67 >-4.00 >-4.00
SR >-4.00 >-4.00
CA 02272275 1999-OS-18 _.
WO 98/23269 PCT/US96/19118
96
Panel/Cell Line Loglo GI50 LogIOTGI Loglo LC50
Ian-Small Cell -
Lung
Cancer -5.82 -4.83 >-4.00
A549/ATCC -4.02 >-4.00 >-4.00
EKVX -6.76 -4.42 >-4.00
HOP~2 i.34 >-4.00 >-4.00
HOP-92 <8.00 -7.23 >-4.00
NCI-H226 -4.41 >-4.00 >-4,00
NCI-H23 -5.42 -4.67 >-4.00
NCI-H322M -6.89 -4.69 >-4.00
NCI-H460 -4.84 -4.21 >-4.00
NCI-H522
Colon Cancer
COLD 20S -5.88 -4.80 -4.35
-
HCC-2998 -7.14 -6.47 -5.58
HCT-116 -6.99 >-4.00 >-4.00
HCT-15 >-4.00 >-4.00 >-4.00
HT29 -6.72 >i.00 >-4.00
KM12 >-4.00 >-4.00 >-4.00
SW-620 >-4.00 >-4.00 >i.00
2 0 CNS Cancer
SF-268 -6.85 -5.05 >-4.00
SF-295 -4.58 >-4.00 >-4.00
SF-S39 -4.07 >-4.00 >i.00
SNB-19 >-4.00 >-4.00 >-4.00
2 5 SNB-75 -4.56 -4.15 >-4.00
U251 -7.13 -4.79 -4.20
WO 98/23269
- -
Panel/Cell Line Loglo GI50LogIOTGI Loglo LC50
Melanoma
MALME-3M -4.35 >-4.00 >-4.00 -
M 14 -4.46 >-4.00 >-4.00
SK-MEL-2 -5:23 -4.61 -4.11
SK-MEL-28 >-4.00 >-4.00 >-4.00 . .
SK-MEL-5 -4.19 >-4.00 >-4.00
UACC-257 -7.56 -6.92 >-4.00
__. . UACC-62 -7.82 -7.29
Ovarian Cancer
IGROV 1 -7.4S i.62 -4.61
OVCAR-3 . _ ..~ -6.19 -4.48 >i.00
_-,
OVCAR-4 i.62 -4.19 >-4.00
OVCAR-5 -7.48 -6.69 -4.56 --
OVCAR-8 _ -4.08 >-4.00 >-4.00.
SK-OV-3 -5.04 >-4.00 >-4.00
Renal Cancer
786-0 _ -4.22 >-4.00 ->-4:00
A498 <-8.00 <-8.00 -7.52
ACHN - -4.28 >-4.00 =.i.00
2 0 RXF-393 >-4.00 >-4.00 >-4.00
SN12C >-4.00 >-4.00 >-4.00
T'K-10 -7.71 r7.12 >-400
UO-31 -4.00 >-4.00 >-4.00
Prostate Cancer
-2 5 - PC-3 -4.26 >-4.00 >-4:00
DU-145 - -4.11 >-4.00 >-4.00
CA 02272275 1999-OS-18
PCT/U896/19118
97
CA 02272275 1999-OS-18
WO 98I23269- - PCT/(TS96/19118
98 '
Panet/Cell Line Loglp GI50 LogIpTGI Loglp LC50
Breast Cancer
MCF7 -5.96 >-4.00 >-4.00
MCF7/ADR-RES -4.34 >-4.00 >-4.00
MDA-MB-231/ATCC -4.51 -4.16 >-4.00
HS 578T >-4.00 >-4.00 >-4.00
IvIDA-IvIB-435 -4.26 >-4.00 >-4.00
MDA-N -4.21 >-4.00 >i.00 - ~~~~-
. . . BT-549 >~.~ ,..~ >.i.00
T-47D -6.59 >-4.00 >-4.00
MG MID -S.30 -4.S6 -4.12
Delta 2.70 3.44 3.39
Rage 4.00 4.00 3.52
_ CA 02272275 1999-OS-18
W0 98/23269 PCT/US96/19118
NSC: 660642-W/0-1/ 11 -
S S S
PanelICel1 Line Loglp GI50L9glp TGI Loglp LC50
Leukemia
CCRF-CEM -5.73 -5.35
HL~O(TB) -5.9l -5.52 -5.12
K-562 -6.19 -5.65 -
MOLT-4 -5.77 -5.45
RPMI-8226 -S.60 -5.16 >-4.00
SR -5.76 -5.44 ~ -5.12
Non-Small Cell Lung
Cancer -5.80 -5.21 -4.44
2 0 A549/ATCC -4.73 -4.39 -4.04
EKVX -5.46 -4.94 -4.22
HOP-62 -5.15 - -4.63 -4.13
HOP-92 -5.46 -4.80 -4.25
NCI-H23 -5.S8 -5.03 -4.52
2 5 NCI-H322M -5.49 -4.72 >-4.00
NCI-H460
CA 02272275 1999-OS-18
WO 98/23269 - - PCT/US96/19118
l00
Panel/Cell Line LoglO GI50Loglo TGI Loglo LC50
Colon Cancer
COLD 205 -5.76 -5.30 -4.12
HCC-2998 -5.60 -4.93 -4.25
HCT-116 -6.37 -S.78 -5.37
- HCT-15 -5.74 -5.40 -5.07
HT29 -5.88 -5.46 -5.04
KM 12 -5.27 -4.67 -4.16
SW-b20 -5.82 -5.14 -4.29
-- - CNS Cancer __
1 0 SF-268 -5.53 -5.05 -4.18
SF-295 -5.07 -4.59 -4.15
SF-539 ~ '-- ~' -4.86 -4.56 -4.27
SNB-19 -5.21 -4.72 -4.36
SNB-75 -5.04 -4.62 -4.21
U251 -5.89 -5.30 - -4.66
- Melanoma
MALME-3M -4.79 -4.44 - -- -4.08
M14 -S.59 -5.13 -4.15
SK-MEL-28 -4.93 -4.58 -4.23
SK-MEL-5 - -5.47 -4.83 .--.i,34
UACC-257 -6.63 -6.29 -5.52
UACC-62 -6.26 -5.12 i.21
Ovarian Cancer
IGROV 1 i.57 -5.91 >-4.00
OVCAR-3 -6.60 -6.17 -5.11
OVCAR-4 -5.72 -5.25 -4:25
OVCAR-5 -6.07 -5.41 -4.74
OVCAR-8 -5.35 -4.59 >-4.00
CA 02272275 1999-OS-18 .
WO 98/23269 PCT/US96/19118
10l
Panel/Cell Line Loglo GI50L.oglo LoBlo LC50
TGI
Renal Cancer
- 786-0 -5.01 -4.S8 -4.16
A498 -6.69 -6.35
CAKI-1 -6.67 -0.25 -5.27
RXF-393 -5.59
i_10 -6.31 -5.27 ~.~
UO-31 -5.27 -4.74 -4.37
Prostate Cancer
PC>3 -5.20 -4.72 -4.34 r
DU-145 -5.61 -5.13 -4.54
Breast Cancer
MCF7 -6.22 -5.45 >-4.00
MCF71ADR-RES -5.55 -5.08 -4.11
MDA-MB-231/ATCC -5.66 -5.19 -4.61
. 15 MDA-N -5.69 -5.34 -5.00
T-47D -5.51 -4.58 >-4.00
_ MG MID -5.67 -5.15 -4.43
Delta 1.02 1.20 1.09
. Range 1.96 1.96 1.52
_ a ~ ~ ~ ~
- CA 02272275 1999-OS-18
WO 98/23269 PCT/LTS96/19118
102
- NSC: 647452-L/1
xoc~ I s I I S ~ c~ox
PaneI/Cell Line LoglO GI50Loglp Loglp LC50
TGI
Leukemia . _.
CCRF-CEM >-4.00 >-4.00 >-4.00
HL-60(TB) _ >-4.00 >-4.00 >-4.00
K-562 -5.22 >-4.00 >-4.00
MOLT-4 >-4.00 >-4.00 >-4.00
RPMI-8226 >-4.00 >i.00 _. >~.~
SR >-4.00 >-4.00 >-4.00
Non-Small Cell
Lung
Cancer -5.91 -5.05 >-4.00
A549/ATCC >-4.00 >i.00 >-4.00
EKVX -5.S1 -4.67 _ >-4.00
2 0 HOP-18 -5.32 -4.21 >-4.00
_. HOPfi2 -4.09 _.-_ .00 >~.~ -
HOP-92 -7.60 -7.13 -b.2I
_ NCI-H226 >-4.00 >-4.00 >-4.00
NCI-H23 -5.77 -5.10 >-4.00
2 5 NCI-H322M i.29 >-4.00 >-4.00
NCI-H460 >-4.00 >-4.00 >-4.00
NCI-H522 -6.33 - -5.66 >-4.00
~_
LXFL 529
Small Cell Lung
Cancer
3 0 DMS 114 >-4.00 >-4.00 >-4.00
DMS 273 -5.22 >-4.00 >-4.00
- -~CA 02272275 1999-OS-18
WO 98I23269 PCT/US96/19118
103
Panel/Cell Line Loglo GI50Loglo TGI Loglo LC50
Colon Cancer
COLD 205 -6. Z2 -5.61-
_ DLD-1 -4.74 >-4.00 >-4.00
HCC-2998 -6.40 -5.45 -4.52
HCT-116 -5.27 >-4.00 >-4,00
HCT-15 -4.19 >-4.00 >-4.00
HT29 -5.04 >-4.00 >-4.00
KM12 -4.35 >-4.00 >-4.00
KM2012 -6.15 -5.4I >-4.00
_ 10 SW-620 -5.04 >-Q1.00 >-4.00
- - CNS- Cancer _____.
SF-268 >-4.00 >-4.00 >-4.00
SF-295 >-4.00 >-4.00 >.4.00 - . ._
. _ SF-539 _ >-4.00 >-4.00 >-4.00
SNB-19 >-4.00 >-4.00 >-4.00
_ SNB-75 _ >:4.00 >-4.00 >-4.00 _
---_.._ _
SNB-78 >-4.00 >-4.00 >-4.00
U25-1- -4.72 >-4.00 >-4.00
XF 498
2 0 Melanoma
LOX IMVI >-4.00 >-4.00 >~.oo
MALME-3M >-4.00 >-4.00 >-4.00 _
M14 >-4.00 >-4.00 >-4.00
M19-MEL >-4.00 >-4.00 >-4.00
2 5 SK-MEL-2 >-4.00 >-4.00 >-4,00
- SK_MEL-28 - - >-4.00 >-4.00 >-4.00
SK-MEL-5 >-4.00- >-4.00 >-4.00
-
UACC-257 -6.00 -5.51
UACC-62 -6.36 -5.60 -4.48
CA 02272275 1999-OS-18
WO 98I23269 PCT/LTS96119118
104
P~el/Cell Line Loglo GI50~8to TGI I-Slo
LC50
Ovarian Cancer
IGROV I -6.57 -5.73 >-4.00
OVCAR-3 -5.70 -5.19 -4.01
OVCAR-4 -4.46 >-4.00 >-4.00
OVCAR-5 -6.06 -5.26 >-4.00
OVCAR-8 -5.23 -4.32 >-4.00
SK-OV-3 -5.80 -S.08 >-4.00
Renal Cancer
786-0 >-4.00 >-4.00 >-4.00
A498 -7.71 -7.36 -7.00
ACHN >-4.00 >-4.00 >-4.00
CAKI-I -0.63 -5.82 >-4.00
~-393 -7.05 >-4.00
RXF-031 >-4.00 >-4.00 >-4.00
SN12C >-4.00 >-4.00 >-4.00
TK-10 -6.29 >-4.00
UO-31 .00 >-4.00 >-4.U0
- MG MID -4.94 -4.46 -4.11
Delta 2.78 2.90 2.89
2 0 _ Range 3.71 3.36 3.00
CA 02272275 1999-OS-18
WO 98/23269 - PCT/US96119118
105 _
NSC: 660644-Y/0-2/12
-
_ ( S ~ ~ s~---CH2~H
y
S
Pane11Ce11 Line Loglo GI50Loglo TGI Loglp LC50
~~e~a _
CCRF-CEM -4.61 > 4.00 > 4.00
HL-60(TB) -4.64 > 4.00 > 4.00
- K-562 -6. 37 > 4.00 > 4: ~ ... .
MOLT-4 -4.49 > 4.00 . _ > 4.00
.
RPMI-8226 -4.22 > 4.00 > 4.00
SR -4.S2 > 4.00 > 4.00
Non-Small Cell Lung
Cancer -4.73 > 4.00 > -4.00 -
2 0 A549/ATCC -4.19 > 4.00 > -4.00
EKVX~ -4.75 -4.27 > ~.~
HOP~2 -4.63 -4. lr > -4.00
HOP-92 -7.85 -6.86 -6.15
__ NCI-H226 -4.43 > -4.00 > ..4.00
2 5 NCI-H23 -4.49 > -4.00 > -4.00
- NCI-H322M ' -4. 87 > -4.00 > -4.00
NCI-H460 . -4.54 > -4.00 > -4.00
NCI-H522
CA 02272275 1999-OS-18
WO 98123269 ~ PCT/US96/19118
106
Panel/CeII Line 81o C'~0 I-81o TGI 81o I-C50
Colon Cancer
COLD 205 -5.93 > -4.00 > -4.00
HCC-2998 -4.88 >-4.00 - >.4.00 _
HCT-116 -4.03 >-4.00
HCT-15 -4.26 > -4.00 > -4.00
HT29 > -4.00 > -4.00
KM12 i.10-- >-4.00 >-4.00
SW-620 -6.S2 > -4.00 > -4.00
CNS Cancer
SF-268 -4.08 >-.4.00 _ >i.00
__ SF_295 __. i.57 -4.09 .00
SF-539 -4.41 >-4.00 >i.00
SNB-19 -4.33 > -4.00 > -4.00
_ _ SNg-75 -4. 83 -4.41 > -4.00
U251 -6.68 -4.59 -4.06
Melanoma
__
LOX-.IMVI -4.42 >-4.00 >-4.00
MALME-3M -4.52 >-4.00 >-4.00
M 14 -4. S0 > -4.00 > -4.00 .
2 0 - SK-MEL-2 -4.69 -4.15 > -4.00 -
SK-MEL-28 -4.67 -4.07 > -4.00
SK-MEL-S -4.54 -4.06 > -4.00
UACC-257 -?.88 -7.39 -6.76
UACC-62 > 4.00 > -4.00
2 5 Ovarian Cancer _
OVCAR-3 i.60. -4.45 >-4.00
OVCAR-4 Q1.48 > -4.00 > -4.00
OVCAR-5 -6.53 -6.09 >-4.00
OVCAR-8 -4.61 >4.00 >-4.00
3 0 SK-OV-3 i.73 -4.27 _
CA 02272275 1999-OS-18
WO 98/2326Q - PCT/US96/19118
107 -
Panel/Cel1 Line Loglo GI50 Loglo TGI Loglo LC50
Renal Cancer
786-0 -4.65 -4.29 _. > .4.00
A498 -7.87 -7.42 -6.93
ACHN -4.14 > 4.00 > -4.00
' 5 CAKI-1 -7.53 i.90 -4.11
_ i_393 -4.49 -4.04 > -4.00
SN 12C > -4.00 > -4.00 > i.00
TK-10 -6.84 >-4.00 _. __
UO-31 >'4.~ ~... >4.00>-4.00
Prostate Caacer
PC-3 -4.5S > -4.00 > -4.00
DU-14S -4.28 >-4.00 >-4.00
Breast Cancer
MCF7 -6.94 ' -4.52 > -4.00
MCF7/ADR-RES -4.39 > 4.00 >
MDA-MB-231 /ATCC -d.60 ~.OS > -4.00
gS 578T - _ -4.54 -4.06 > -4.00
MDA-MB-435 -4.45 > -4.00 > i.00
MDA-N -4.54 >-4.00 >-4.00
BT 549 > -4.00 > -4.00 > -4.00
T-47D -6.67 >-4.0p
MG MID ~ -
Delta -5.05 -4.32 -4.14
Range 2.84 3.10 2.80
3.88 3.42 2.93
CA 02272275 1999-OS-18 -.
WO 98I23269 PCT/US96/19118
108
NSC: 663561-U/0-1/17
CHZOH
j
~ CH2~
Panel/Cel1 Line LoglO GI50LoglO TGI Loglp LC50
Leukemia
CCRF-CEM -5.S3 >-5.00 >-5.00
HLfiO(TB) -5.51 >-5.00 >-5.00
K-562 -7.07 >-5.00 >-5.00 . _
MOLT-4 -5.61 >-5.00 >-5.00
SR -5.29 >-5.00 >-5.00
Non-Small Cell Lung __
Cancer -6.33 >-5.00 >-5.00
A549/ATCC >-5.00 >-5.00 >-5.00 -
EKVX >-5.00 >-5.00
2 0 HOP-62 -5.23 >-5.00 >-5.00
_ HOP-92 -7.75 -7.29 -6.59
NCI-H226 -5.l0 >-5.00 >-S.00
NCI-H23 >-5.00 >-5.00 >-5.00
-- NCI-H322M -7.28 >-5.00 >-5.00
2 5 NCI-H460 -5.03 >-5.00 >-5.00
NCI-H522
CA 02272275 1999-OS-18
WO 98I23269 - - " - PCT/US96/19118
109
Panel/Cell Line Loglo GI50Loglo TGI Loglo LC50
Colon Cancer
' HCC-2998 -S.63 >-5.00 >-5.00
_ HCT-116 -7.35 >-5.00 >-5.00
HCT-15 -5.11 >-5.00 >-5.00
-6.57 >_S.00 >-5.00
KM12 -5.17 >-5.00 >-5.00
SW-b20 -7.53 >-5.00 >-5.00
CNS Cancer
SF-268 >-5.00 >-5.00 >-5.00
SF-29S > S.00 >-S.00 >-5.00
SF-539 >-5.00 >-5.00 >-5.00
SNB-i9 - .~ ~-~ >_5.00 >-5.00 >-5.00
SNB-75 -5.26 >-5.00 >-5.00
U251 -5.60 >-5.00 >-5.00
Melanoma -
LOX IMVI -5.47 >-5.00 >-5.00
MALME-3M >-5.00 >-5.00 ' >-5.00
M14 >-5.00 >-5.00 >-5.00
SK-MEL-2 -5.64 >-S.00 >-5.00
2 0 SK-MEL-28 - >-5.00 >-5.00 >=5.00
SK-MEL-5 -5.31 >-5.00 >-5.00
UACC~2 -7.68 >-5.00 >-5.00
Ovarian Cancer
IGROV 1 -8.05 -7.06 >-5.00
2 5 OVCAR-3 -b.74 >-5.00 >-S.00
OVCAR-4 i.27 >-5.00 >-S.00
OVCAR-5 -7.11 -6.45 >-5.00
_ OVCAR-8 - >.5.00 >-5.00 >-5.00
CA 02272275 1999-OS-18
WO 98I23269_ - PCT/US96/19118
l10
Panel/Cell Line I-81o GI50I-a8lo L81o L~
TGI
Renal Cancer
786-0 >-5.00 >-5.00 >_5.00
A498 -7.85 -7.50 -7.16
ACHN >-5.00 >-5.00 >-5.00
CAKI-1 -7.63 -7.05 >-5.00
RXF-393 -5.57 >-5.00 >-5.00
~SN12C -5.46 >-5.00 >-5.00
TK-10 -7.49 -6.78 >-5.00 - '
UO-31 >-5.00 _._>_5.00 >-5.00
Prostate Cancer
PC-3 -5.06 >-5.00 >-5.00
DU-145 >-5.00 >-5.00 >-5.00
Breast Cancer
MCF7 -7.00 -6.05 >-5.00
MCF7/ADR-RES >-S.00 >-5.00 >-5.00
MDA-MB-435 -5.25 >-5.00 >-5.00
-
MDA-N -5.47 >-5.00 >-5.00
BT-549
MG MID -5.84 -S.25 -5.07
2 0 Delta 2.21 2.25 2.09
Range 3.05 2.50 2.16
CA 02272275 1999-OS-18
WO 98/Z3269 - PCT/US96I19118
111
NSC: 663562-V/O-1/18
Pane1lCell Q810
Line Loglo GI50Loglo TGI LC50
_ 10 Leukemia __
CCRF-CEM -6.08 -5.14 >-5.00
HL~O(TB) -5.36 >-S.00 >-5.00
K-562 -6.84 -5.86 -S.03
MOLT-4 -6.11 -5.14 >-5:00
15 SR -5.61 >-5.00 >-5.00
Non-Small Cell Lung
C~~r -6.34 -5.69 _5.25
A5491ATCC -5.89 -5.57 -5.25
EKVX -6.68 -5.65 -5.10
2 0 HOP-62 -5.92 . -5.53 -5.14
HOP-92 -7.87 -7.28 -6.34
NCI-H226 -5.86 - -5.48 -5.10
NCI-H23 -5.74 -5.42 -5.11
NCI-H322M -7.14 -5.92 -5.39
2 5 NCI-H460 -5.76 -5.49 -S.22
NCI-H522
Colon Cancer
- HCC-2998 -5.74 -5.42 -5.10
- HCT-116 -7.33 -5.87 -5.43
3 0 HCT-15 -5.80 -5.37 >-5.00
HT29 -6.54 -5.89 -5.41
KM12 -5.84 -5.52 -5.19
SW~20 -7.07 -5.79 _ >.5 00 _
CA 02272275 1999-OS-18
WO 98/23269 - l PCTIUS96/19118
1l2
Panel/Cell I-g1o
Line Loglp GISOLoglp TGI LC50
FNS Cancer
SF-268 -5.76 -5.42 -5.09
SF-295 -5.83 -5.49 -5.16
SF-539 -5.76 -5.48 -5.20
SNB-19 -S.70 -5.31 >-5.00
SNB-75 -5.90 -5.39 >-5.00
U251 -S.96 -5.58 -5.20
Melanoma
LOX IMVI -5.88 -S.49 -5.10
MALME-3M -5.75 -S.40 -S.06
M14 -5.69 -5.40 -5.11
SK-MEL-2 -5.81 5.19 >-5.00
SK-MEL-28 -5.75 -5.45 -5.15
SK-MEL-5 -5.88 -S.58 ' -5.28
UACC-62 -7.82 -6.59 -5.10
Ovarian Cancer
IGROV 1 - -7.68 -6.68 -5.61
OVCAR-3 -6.85 -6.07 -5.08
OVCAR-4 - -6.2S -5.59 __-_5.01
2 0 OVCAR-5 -6.93 i.30 -5.59
OVCAR-8 -5.77 -5.49 -5.21
Renal Cancer
786-0 -5.94 -5.61 -5.29
A498 --7.76 -7.37 -6.92
2 5 ACHN -5.77 -5.51 -5.25
CAKI-1 -7.41 -6.50 >-5.00
_ RXF-393 -6.26 -5.64 -5.18
SN12C -5.92 -S.52 -S.11
TK-10 -6.93 -6.27 _ >-5.00
3 0 UO-31 -5.72 -5.48 -5.23
- CA 02272275
1999-OS-18
WO 98/23269 PCT/US96/19118
113
Panel/Cell L810
.. Line Loglo GI50 Loglo LC50
TGI
Prostate Caacer_
PC-3 -S.63 >-5.00 >-S.00
DU-145 -S.66 -S.18 >-S.00
Breast Cancer
MCF7 -6.61 -6.00 -5.32
MCF7/ADR-RES -S.43 -S.OS >-S.00
~
MDA-MB-43S -S.77 -S.47 -S.18
_ MDA-N - _5.77 - "'-S':45-5.13 -
BT-S49 -5.62 -5.25 >-S.00
1 0 MG MID -6.20 -S.65 -S.20
Delta 1.67 1.72 1.72
Range 2.S2 2.37 --- 1.92
CA 02272275 1999-OS-18
WO 98I23269 PCT/US96/19118
114
NSC: 666165-Z f 0-1/30 -
os
Panel/Cel1 Line Loglp GI50Loglo TGI 81o L~0
Leukemia
ccRF-c~M -4.ss -4.16 >.~.~
=HL~O(TB) -4. 36 > -4.00 > .d.00 -
K-562 -5.23 > -4.00 > -4.00
MOLT-4 -4.63 -4.18 >-4.00
RPMI-8226 -4.63 -4.11 >-4.00
SR -4.64 -4.01 > -4.00
Non-Small Cell Lung
Cancer -5.58 >-4.00 >-4.00
2 0 A549lATCC -4.74 -4.02 > -4.00 -
EKVX -4.65 -4.20 >-4.00
HOP-62 -5.37 -4.40 > -4.00
HOP-92 -6.64 -5.42
NCI-H226 -4.67 -4.27 > -4.00
2 5 NCI-H23 -5.71 -5.24
NCI-H322M -6.32 -5.69 -5.17
NCI-H460 -4.91 -4.58 -4.24
NCI-H522
CA 02272275 1999-OS-18
WO 98/23269 PCT/US96/19118
115
Panei/Cell Line Q810 r'~0 I-810 TGI I-81o I-C50
Colon Cancer
COLO 205 -5.78 - -5.17 > -4.00
HCC-2998 -4.53 - -4.03 > i.00
HCT-1 I6 - -4.91 -4.43 > -4.00
HCT-15 -4.59 -4.04 >-4.00
HT29 -4. 88 -4.33 > -4.00
- KM 12 -4.75 -4.46 -4.17
SW-620 -4.~h > -4.00 > -4.00
CNS Cancer
' F SF-268 i.55 -4.04 > 4.00
SF-295 -
SF-S39 -4.69 i.39 -4.09
SNB-19 - -4.44 > 4.00 > -4.00
SNB-75 -4.64 -4.15 > -4.00
U251 -6.23 -4.30 > -4.00
Melanoma
LOC IMVI. -4.60 -4.11 >-4.00
- MALME-3M -4.71 -4.32 >-4.00 -
M 14 -4.76 -4.50 -4.23
2 0 SK-MEL-2 -4.69 =4.31 > 4.00
SK-MEL-28
SK-MEL-5 _ . -4.82 -4.53 - -4.24
UACC-257 -7.66 -7.15 -6.54
UACC-62 -7.48 i.15
2 5 Ovarian Cancer
IGROV1 -7.07 -5.64 >-4.00
OVCAR-3 i.61 -4.39 >-4.00
OVCAR-4 -4.74 -4.17 >4.00
OVCAR-5 -6.44 __. -6.05 >-4.00
3 0 OVCAR-8 -5:2 T -4.72 -4.34
SK-OV-3 -5.70 -4.80 >-4.00
I I I I I
CA 02272275 1999-OS-18
WO 98I23269 PCT/US96/19118
116
PanellCell Line Loglo GI50Logla Loglp LC50
TGI
Renal Cancer . . _
786-0 - -4.60 > -4.00 > -4.00
ACHN -4.55 > -4.00 > -4.00
CAKI-1 -7.61 > -4.00
RXF-393 -4.72 -4.28 > -4. 00
-
SN 12C -4.58 -4.03 > -4.00
-- TK-10 -6.42 -4.63 >-4.00
,
- UO-31 -4.71 -4.39 -4.07
_ prostate Cancer _ - -
PC-3 -4.72 -4.24 > -4.00
DU-145 -4.88 -4.47 -4.06
Breast Cancer
MCF7 -7.47 -5.38 -4.28
MCF71ADR-RES -5.04 >-4.00 >-4.00
MDA-MB-231/ATCC -4.58 -4.07 >-4.00
HS 578T -4.42 > -4.00 > -4.00
MDA-MB-435 -4.62 -4.25 > -4.00
MDA-N - -4.73 -4.38 - -4.03
.BT-549 -4.66 -4.30 > -4.00
2 0 T-47D -4. 85 -4.43 -4.01
- MG MID
Delta -5.21 -4.46 -4.10
Range 2.45 2.69 2.43
3.30 3.15 2.54
CA 02272275 1999-OS-18
WO 98/23269 - PCT/I1S96/19118
117
NSC: 65811Z-W/0-1/20
_ ~ZOH __ .
s s s
Panel/Cell Line Loglp GI50 Loglp TGI Loglp
LC50
Leukemia
CCRF-CEM -4.02 > -4.00 > -4.00
_
HL-60(TB) -4.70 > -4.00 > -4.00
K-562 -4.61 > -4.00 > -4.00
MOLT-4 > -4.00 > -4.00 > -4.00
RPMI-8226 > -4.00 > -4. 00 > -4.00
Non-Small Cell Lung
Cancer -4.60 > -4.00 > -4.00
A549/ATCC -4.29 > -4.00 > i.00
HOP-62 > -4.00 > -4.00 > -4.00
HOP-92 -4.16 > -4.00 > -4.00
2 0 NCI-H226 -4.39 > -4.00 > -4.00
NCI-H23 -4.41 >-4.00 >-4.00
NCI-H322M -4.56 > -4.00 > -4.00
NCI-H460 -4.10 > i.00 > -4.00
__. NCI-H522
2 5 Colon Cancer
COLO 205 -4.72 -4.48 -4.24
- HCC-2998 -4.75 -4.35 >-4.00
HCT-116 -4.41 > ~.~ > i.00
HCT-15 -4.60 > -4.00 > i.00 _
3 0 HT29 -4.25 > -4.00 >_i.00
KM12 -4.90 -4.23 > -4.00
_ SW~20 -4.45 > -4.00 > -4.00
CA 02272275 1999-OS-18
WO 98/23269 PCT/US96/19118
118
Panel/Cell Line Loglo GISOLogy TGI Logy LC50
CNS Cancer -
SF-268 -4. 36 > -4.00 > -4.00
SF-295 -4.50 > -4.00 _ > -4.00
SF-539 -4.63 -4.17 > -4.00
SNB-19 > i.00 > 4.00 > -4.00
SNB-75 -4.56 > 4.00 > -4.00
U251 -4. 39r- > 4.00 > -4.00
Melanoma
- LOX IMVI -4.60 >-4.00 .00
MALME-3M -4.55 >-4.00 >-4.00
_ . M 14 --_. -4.43 > -4.00 > -4.00
SK-MEL-2 -4.40 > i.00 > i.00
SK-MEL-28 -4.39 > -4.00 > -4.00
SK-MEL-5 -4.47 > -4. 00 > -4.00
UACC-257 -4.51 > -4.00 > -4.00
- UACC-62 -4.71 > -4.00 > -4.00
Ovarian Cancer
IGROV 1 -4.18 > -4.00 > -4.00
OV CAR-3 -4.44 > -4.00 > -4. 00
2 0 - OVCAR-4 > -4.00 > -4.00 > -4.00
OVCAR-5 -4.38 >-4.00 > i.00
OVCAR-8 > 4.00 > -4.00 > -4.00
SK-OV-3 -4.07 > -4.00 > -4.00
Renal Cancer -
2 5 786-0 > -4.00 > -4.00 > -4.00
A498 -4.61 .- -4.05 > -4.00 -
ACIiN -4.43 > -4.00 > -4.00
CAKI-1 -4.19 >-4.00 >-4.00
RXF-393 > -4.00 > -4.00 > -4.00
3 0 TK-10 -4. 39 > -4.00 > -4.00
UO-31 > 4.00 > -4.00 > ~. ~
CA 02272275 1999-OS-18
WO 98I23269 - PCT/US96/19118
119 -
Panel/Cell Line Loglo GI50 Loglo TGI 81o LC50
Prostate Cancer
pC_3 -4.39 > -4.00 > -4.00
DU-145 -4.13 >-4.00 >-4.00
Breast Cancer
- 5 MCF7 -5.45 -4.43 > -4.00
MCF7/ADR-RES -4.39 >-4.00 >-4,00
V
MDA-MB-2311ATCC i.49 > -4.00 > -4.00
HS 578T -4.53 -4.41 > -4.00
-. . .. _ MDA - .. .. i.41> -4.00
MB-435 .i.85
MDA-N -5.09 -4. S 1 > -4.00
BT-549 -4.52 i.04 > -4.00
T-47D -5.17 -4.40 > -4.00
MG MID
Delta -4.42 -4.05 -4.00
- Range 1.02 0.46 0.24
1.45 0.51 0.24
CA 02272275 1999-OS-18
WO 98I23269 PCT/US96/19118
120
NSC: 658878-G/0-1/16
c&oH
s s s
Panel/Cell Line ~ Loglo GI50Loglp TGI Loglo
LC50
Leukemia
I 0 CCRF-CEM -4.45 > -4.00 > -4.00
L.u,~O~B) -4.78 >-4.00 >-4.00
K-S62 -6.44 > -4.00 > -4.00
MOLT-4 -4.9I > -4.00 > i:00 .. _
RPMI-8226 -4.50 > -4.00 > -4.00
15 Non-Small Cell Lung _
Cancer -4.44 >-400 >-4.00
A549/ATCC -4.24 > -4.00 > -4.00
EKVX -4. 70 > -4.00 > -4.00
HOP~2 > -4.00 > -4.00 - > -4.00
2 0 HOP-92 -6.59 i.09 > i.00
NCI-H226 -4.41 > -4~0 > -4.00
NCI-H23 -5.25 -4.32 >-4.00
NCI-H460 -4.47 >-4.00 >-4.00
NCI-H522
2 5 Colon Cancer
COLD 205 -5.23 -4.55 -4.00
HCC-2998 -6.50 -4.?2 -4.11
HCT-15 -4.55 > -4.00 > -4.00
HT29 -4.73 > i.00 > -4.00
3 0 KM 12 ~- -4.78 -4.10 > -4.00
SW~20 -4.39 >-4.00 >-4.00
CA 02272275 1999-OS-18
WO 98/23269 - " PCT/U896/19118
121
Panel/Cel1 Line 81o GI50 Loglo TGI 81o LC50
CNS Cancer
SF-268 > -4.00 > -4.00 > -d.00
SF-295 -4.47 > -4.00 > -4.00
SF-539 > -4.00 > -4.00 > -4.00
SNB-19 > -4.00 > -4.00 > -4.00
U251 -4.16 > .i.00 > -4.00
Melanoma
MALME-3M -4.53 > -4.00 > -4.00 -
.. M 14 -4.29 > -4.00 > -4. Q0
-
SK-MEL-28 -4.40 > -4.00 > -4.00
SK-MEL-5 -4:46 >-4.00 >-4.00.
UACC-62 > -4.00 > -4.00
Ovarian Cancer
IGROV 1 -6.64 > -4.00 > -4.00
OVCAR-4 _ > -4.00 > -4.00 > -4.00
OVCAR-S >-4.00 >-4.00
OV CAR-8 -4.30 > -4.00 > -4.00
Renal Cancer
2 0 786-0 - -4.28 > -4.00 _a -4.00
ACHN -4.25 > -4.00 > -4.00
RXF-393 -5.06 > -4.00 > -4.00
SN12C -4.33 >-4.00 .00
- TK-10 -4.68 > -4.00 > -4.00
2 5 UO-31 -4.19 > -4.00 > -4.00
Prostate Cancer
PC-3 -4.31 > -4.00 > -4.00
DU-145 -4.26 > -4.00 > 4.00
CA 02272275 1999-OS-18
WO 98l23269- PCT/US96/19118
122
Panel/Cell Line Loglo GI50Loglo TGI Loglo LC50
Breast Cancer
MCF7 -6.75 > -4.00 > i.00
MCF7/ADR-RES -4.24 >-4.00 >-4.00
HS 578T -4.28 > -4.00 > -4.00
- 5 MDA-MB-433 ~ ~. 82 -4.15 > -4.00
MDA-N -5.27 -4.71 -4.24
BT-549 -4.08 > -4.00 > -4.00
~
T-47D i.37 > -4.00 > ~.~ .m- -_
MG MID
Delta -4.73 -4.I0 -4.01
Range 2.01 2.00 0.23
2.75 2.09 0.24
_ CA 02272275 1999-OS-18
WO 98/23269 PCT/US96/19118
123
NSC: 652866-U/1
I I I 4
s s s
Panel/Cell Line Loglo GI50Logio TGI Loglo LC50
Leukemia
CCRF-CEM -5.71 -- -5.36 -5.01
HL-60(TB) -5.84 -5.49 - -5.13
K-562 -5.67 -5.24 -4.66
MOLT=4- - -5.38 -4.73 -4.23
- RPMI-8226 - -5.52 -4.99 -4.45
SR -5.64 -5.13 -4.52
Non-Small Cell Lung -- ---
Cancer -4.74 -4.28 > -4.p0 -
A549/ATCC -4.67 - -4.28 >-4.00
EKVX
2 0 HOP-18 -4.90 -4.56 -4.22
HOP-62
HOP-92 -6.91 _ -6.46 -b.00
NCI-H226 -4.78 -4.51 -4:24
N CI-H23 -5.15 -4.7 I -4.36
2 5 NCI-H322M -6.19 -4.82 -4.34
NCI-H460 -4.86 -4.55 -4.25
NCI-H522 -7.26 -6.61
LXFL 529
Small Cell Lung Cancer
3 0 DMS 114 -5.4I -4.67 -4.29
DMS 273 -4. 80 > -4.00 > -4.00
CA 02272275 1999-OS-18
WO 98I23269 - - ~ PCT/US96/19118
-' 12 4
PanellCetl Line Loglo GI50~.oglo LoglO LC50
TGI
Colon Cancer
COLD 205 -5.39 -4.77 -4.38
_ . DLD-1 -5.33 -4.77 -4.33
HCC-2998 -5.89 -4.99 -4.49
HCT-116 -6.19 -4.70 -4.29
HCT-15 -4.88 -4.54 -4.20
HT29 -5.26 -4.63 i.11
KM 12 -4.78 -4.45 -4.13
KM20L2 -5.79 -4.92 -4.4~
SW-620 -6.40 -4.94 > -4.00
CNS Cancer
/
SF-268 -4.76 -4.46 4.15
SF-295 -4.70 -4.34 > 4.00
SF-539 -4.76 -4.47 -4.18
SNB-19 -4.85 -4.52 ~ -4.19
SNB-75
SNB-78
U251 _ -6.64 -5.79 - -5.15
XF 498 -6.52 -6.05 -5.38
2 0 Melanoma - _ _.
LOX IMVI -4.14 > -4.00 > -4.00
MALME-3M -5.22 -4.72 -4.36
M 14 > -S.00 > -S.00 > -5.00
M 19-MEL -4.82 -4.55 -4.27
2 5 SK-MEL-2 -
- SK-MEL-28 -4.86 -4.S7 -4.29
SK-MEL-5 i.24 -4.56 -4.28
UACC-257 i.60 -6.17 > 5.00
- UACC-62 -7.28 -6.41 -451 _
- CA 02272275
1999-OS-18
WO 98/23269 PCT/US96119118
125 -
Panel/Cell Line, Lo8lo GI50 ~8to TGI I-81o L~0
Ovarian Cancer . _...
' IGROV1 - -6.59 -6.05 -4.38
OVCAR-3 i.54 -4.74 -4.15
OVCAR-4
OVCAR-5 -b.74 i.09 -- -5.12
OVCAR-8 > -S.00 > -5.00 > -5.00
- -- SK-OV-3 -4.76 -4.49 -4.23
:
- Renal Cancer _ _. -- -
786-0 -5.01 -4.67 -4.33
A498 -0.43 -6.24 -6.05
ACHN -4.88 -4.59 i.29
CAKI-1 -6.63 -6.21 -4.75
RXF-393 -5.24 -4.68 -4.29
RXF-631
SN12C -4.83 -4.54 -4.26
TK-10 -6.S8 -6.12 -4.62
UO-31 -4.91 -4.60 -4.29
MG MID --
_ - Delta- -5.52 -4.99 -4.47
2 0 Range 1.76 1.62 1.58
3.14 2.61 2.05
CA 02272275 1999-OS-18
WO 98/Z3269 ~ PCT/US96/19118
126
NSC: 647455-O/1
I I I I i ~o __
s s s
Panel/Cell Line Loglo GI50Loglo TGI LoglQ LCSO
Leukemia
CCRF-CEM > ~.~ _ > ~.~ > i.00
HL-60(TB) > -4.00 > -4.00 - > -4.00
K-562 -5.68 > -4.00 > -4.00
MOLT-4 .. > ~.~ > i.00 > -4.00
RPMI-8226 > -4.00 > -4.00 >-4.00
SR > -4.00 > -4.00 > -4.00
Non-Small Cell Lung - a
Cancer > i.00 > -4.00 > i.00
A549/ATCC > -4.00 > -4.00 > -4.00
EKVX > ~.~ > i.00 >-4.00
2 0 HOP-18 > -4.00 > -4.00 > -4.00
HOP-62 > -4.00 > -4.00 > -4.00
HOP-92 -7.62 - -7.02 -6.28
NCI-H226 > -4.00 > -4.00 > -4.00
NCI-H23 -4.21 > -4.00 > -4.00
2 5 NCI-H322M i.88 > -4.00 > -4.00
NCI-H460 -4.66 -4.34 -4.02
NCI-H522 -7.44 -6.78
LXFL S29
Small Cell Lung Cancer
30 DMS 114 >-4.00 >-4.00 >-4.00
DMS 273 > -4.00 > -4.00 > -4.00
CA 02272275 1999-OS-18
WO 98/23269 PCT/U596/19118
127
Panel/Cell Line Logy GI50 Loglo TGI Loglo
LC50
Colon Cancer
COLD 205 -5.30 -> -4.00 > -4.00
DLD-1 > -4.00 ~ -4.00 > -4.00
HCC-2998 =4. 31 > -4.00 > -4.00
_ 5 HCT-116 >-4,00 >-.4,00
HCT-15 -4.21 > -4.00 > -4.00
HT29 > -4.00 > -4.00 > -4.00
KM 12 > -4.00 - > -4.00 > -4.00
KM20L2 -6.53 >-4.00 >-4;00
- -- SW-620
CNS Cancer
SF-268 > -4.00 > -4.00 > -4.00
SF-295 ._ ~ > -4.00 > -4.00 > -4.00
SF-539 > -4.00 > -4.00 > -4.00
SNB-19 > -4.00 > -4.00 > -4.00
SNB-75 -4.18 >-4.00 >i.00
SNB-78 > -4.00 > -4.00 > -4.00
U251 . > -4.00 > -4.00
XF 498
2 0 Melanoma
LOX IM V I > -4. 00 > -4.00 > -4.
00
MALME-3M _ >-4.00 >-4.00 ->-4.00
__ M 14 > -4:00 > -4.00 > -4.00
M 19-MEL > -4.00 > -4.00 > -4.00
2 5 SK-MEL-2 > -4.00 > -4.00 > -4.00
SK-MEL-28 > -4.00 > -4.00 > -4.00
SK-MEL-5 > -4.00 > -4.00 > -4.00
UACC-257 -b.73 i.39 -6.04
UACC-62 -7.24 > -4.00
- CA 02272275
1999-OS-18
WO 98I23269 PCTIUS96/19118
l28
Panel/Cell Line Loglo GI50 Loglo Loglo LC50
TGI
Ovarian Cancer . _
IGROV 1 i.83 > -4.00
OV CAR-3 -6.67 > -4.00
OVCAR-4 > -4.00 > -4.00 > -4.00
OV CAR-5 ~. 35 > -4.00
OVCAR-8 > -4.00 > 4.00 > -4.00
SK-OV-3 -4.39 > 4.00 > -4.00
.
Renal Cancer
_ 786-0 >-4.00 >-4.00 .~ -
A498
ACHN > -4.00 > -4.00 > -4.00
C~-1 -7.21 -6.44 > -4.00
RXF-393 < -8.00 > -4.00
RXF-031 -4.02 > -4.00 > -4.00
SN 12C > -4.00 > -4.00 > -4.00
TK-10 -6. 86 > -4.00
UO-31 > -4.00 > -4.00 > -4.00
MG MID -
Delta -4.75 -4.21 -4.08
2 0 Range 3.25 2. 81 2.21
4.00 3.02 2.28
CA 02272275 1999-OS-18
WO 98/Z3Z69 PCT/US96/19118
129
NSC: 637388-V/1
~ ~ ~ ~ ~~20H _ _
S S S
Panel/Cell Line Loglo GISOLoglo TGI Loglo
LCSO
Leukemia
CCRF-CEM > -4.00 > -4.00 > -4.00
HL~O(TB) _ > -4.00 > -4.00 > -4.00
K-562 -6.06 > -4.00 > -4.00
__ MOLT-4 > -4.00 > -4.00 > ~. pp
RPMI-8226 > -4.00 > -4.00 > i.00
SR > -4.00 > -4.00 > ~. ~ - .
Non-Small Cell Lung
~~r > ~.p0 > i.00 >.d.00
A549/ATCC > -4.00 > -4.00 > -4.00
EKV?~ -4.65 > -4.00 > -4.00
2 0 _ _ HOP-18
HOP-92 -6.93 -6.40 -5.S5
_ NCI-H226 -4.15 >-4.00 >-4.00
NCI-H23 -4.53 > -4.00 > -4.00
NCI-H322M -6.31 > -4.00 > -4.00
2 5 NCI-H460 < -8.00 < -8.00
LXFL-529L
Small Cell Lung Cancer
DMS 114 > 4.00 - > -4.00 > -4.00
DMS 273 > -4.00 > -4.00
CA 02272275 1999-OS-18
WO 98I23269 PCT/US96/19118
__ _ 130
Pane11Ce11 Line Loglp GI50Loglp TGI Loglp
LC50
Colon Cancer _
COLD 205 -6.04 > -4.00 > -4.00
DLD-1 -5.0S > -4.00 > -4.00
HCC-2998 i.44 > -4.00 > -4.00
HCT-116 >-4.00 >-4.00
HC'T-15 - > -4.00 > -4.00 > -4.00
HT29 > -4.00 > -4.00 > -4.00
KM 12 > -4.00 > -4.00 > -4.00
KM20L2 > -4.00 > -4.00
SW-620 > i.00 > -4.00
CNS Cancer
SF-268-- - > -4.00 > -4.00 > -4.00
SF-295 > -4.00 > -4.00 > -4.00
SF-539 > -4.00 > -4.00 > -4.00
SNB-19 > -4.00 > -4.00 > -4.00 -
SNB-75 -4. 47 > -4.00 > -4.00
SNB-78 ~ > i.00 > -4.00 > -4.00
U251 > -4.00 > -0.00 > -4.00
XF 498 > -4.00
2 0 Melanoma
LOX IMVI >-4.00 >-4.00 >-4.00
_- MIME-3M > -4.00 > -4.00 > -4.00
M 14 > -4.00 > -4.00 > -4.00
M i-9-MEL > -4.00 > -4.00 > -4.00
2 5 SK-MEL-2 > -4.00 > -4.00 > -4.00
- SK-MEL-28 > -4.00 > -4.00 > -4.00
SK-MEL-5 > -4.00 > -4.00 > -4:00
UACC-257 i.99
UACC~2 <-8.00 __ _2.85
CA 02272275 1999-OS-18 _
WO 98/Z3269 - PCT/U596/19118
l31
P~el/Cell Line Loglo GI50 Q810 TGI ~Slo L~0
Ovarian Cancer
IGROV 1 -7.37 > -4.00
- OVCAR-3 _ > -4.00 > -4.00
OVCAR-4 > -4.00 > -4.00 > -4.00
OVCAR-5 -6.65 >-4.00 >-4.00
OVCAR-8 > -4.00 > -4.00 > -4.00
SK-OV-3 > -4.00 > -4.00 > -4.00
Renal Cancer
786-0 > -4.00 > -4.00 > -4.00
A498 < -8.00 -7.49 -6: 80
ACHN > -4.00 > i.00 > -4.0o _
RXF-393 -7.56 > -4.00 > -4.00
RXF-631 -7.50 ~. i l > -4.00 . _
SN 12C -7.55 > -4.00
TK-10 > -4.00 > -4.00 > -4.00
_
UO-31 -6.98 > -4.00
> -4.00 >=4:00 > -4.00
MG MID
Delta -4.39 -4.30 -4.08
Range 3.07 3.70 2.72
4.00 4.00 2.80
CA 02272275 1999-OS-18
WO 98I23269 PCT/US96I19118
I32
NSC: 637393-A/1
I _
CH4
S S S
Panel/Cel1 Line Loglo GISWLoglo TGI Loglo LC50
Leukemia
CCRF-CEM -5.50 -4.68 > -4.00
- _ HL-60(TB) -S.82 -5.25 - > -4.00
K-562 -6.09 -4.45 > -4.00
MOLT-4 -S.53 > -4.00 > -4.00
RPMI-8226 -5.31 -4.09 > -4.00
I5 SR -5.58 -4.62 - >-4.00
- Non-Small Cell Lung
_ _ Cancer -4.73 -4.36 > -4.00
A549/ATCC -4.71 -4.25 > -4.00
- - - EKVX -5.05 -4.32 >-4.00
2 0 HOP-18 i.03 -4.65 ~> -4.00
HOP-92 -6.69 -6.14 -5.23
NCI-H226 -4.69 -4.21 > -4.00
NCI-H23 -4.78 -4.36 > -4.00
NCI-H322M -5.70 -4.65 -4.12
2 5 NCI-H460 - < -8.00 < -8.00
LXPL-529L
Small Cell Lung Cancer
DMS 114 -4.79 -4.24 >-4.00
DMS 273 -4.94 -4.55 -4.16
CA 02272275 1999-OS-18
WO 98/23269 - - - - PCT/US96/19118
133
I'anel/Cell Line Loglo GI50 Loglo Loglo LC50
TGI
Colon Cancer
COLO 205 -5.37 -4.69 -4.Z5
- DLD-1 -5.13 -4.47 > -4.00
HCC-2998 -4.86 -4.49 -4.12
HCT-116 -7.04 -4.S3 >-4.00
H_15 -4.87 -4.45 -4.03
HT29 -4. 72 > -4.00 > -4.00
KM12 -4.75 -4.28 > -4.00
KM20L2 i.28 > -4.00 > -4.00
SW~20 - -6.37 >-4.00 >-4.D0
CNS Cancer
SF-268 -4.59 > -4.00 > -4.00
SF-295 _ ~-- -.~ ~.~ .4.07 > ~.~
-
SF-S39 -4.76 -4.34 >-4.00
SNB-19 -4.62 >-4.00 >-4.00
SNB-75 -5.30 -4.53 ~ > -4.00
SNB-78 -4.82 -4.28 >-4.00
U251 -4.88 > -4.00 , ._ >
-4.00
XF 498 _ ~.~ >-4.00
2 0 Melanoma
LOX IMVI - -4.60 -4.04 __-> ~.~
MALME-3M -4.66 -4.16 > -4.00
M 14 i.67 > -4.00 > -4.00
M 19-MEL -4.44 > -4.00 > -4.00
2 5 SK-MEL-2 -4.70 -4.35 > -4.00
SK-MEL-28 -4.72 -4.20 > -4.00 -
- SK-MEL-5 -4.60 > -4.00 > -4. 00
UACC-257 -7.28 > -4.0p -
UACC-62 < -8.00 < -8.00
CA 02272275 1999-OS-18
WO 98/23269_ PCT/US96/19118
13 4 .._
Panel/Cell Line I-Q810 Logio TGI Lo810 L~0
GI50
Ovarian Caacer
- IGROV 1 -7.38 -4.80 - > -4.00
OVCAR-3 -7.21 >-4.00 >-4.00
OVCAR-4 -4.74 -4.06 >-4.00
OVCAR-5 -6.66 -4.30 >-4.00
OVCAR-8 -4.62 -4.01 > -4.00
.SK-OV-3 -4.64 -4.33 -4.02
. . . Renal Cancer
786-0 -4.83 -4.44 -4.05 -
1 0 A498 -7.22 -6.72 -6.29
ACHN -4.6I -4.23 > -4.00
CAKI-I =7.10 -4.72 > -4.00
RXF-393 < -8.00 -4.69 -4.16
RXF-631 -7.68 l -4.47
SN 12C -4.64 -4.10 > -4.00 -
TK-10 -6.53 -4.81 -4.20
UO=31 -4.94 -4.34 > i.00
MG MID -5.53 -4.53 -4.09
Delta 2.47 3.47 2.20
2 0 Range 3.56 4.00 2.29
CA 02272275 1999-OS-18
W098/23269 PCT/I1S96/19i18
- 135
NSC: 647455-O/1
Hoa2c I I I I I I ~
. s s s
Panel/Cell Line ~ Loglo Loglo TGI Loglo LC50
GI50
Leukemia _
CCRF-CEM > -4.00 > i.00 - > i.00
HL~O(TB) > -4.00 > -4.00 > -4.00
K-562 - _ - -5.68 > a1.00 > -4.00
-MOLT-4 > -4.00 > -4.00 > -4.00
RPM I-8226 > -4.00 > -4.00 > -4.00
SR > -4.00 > -4.00 > -4.00
Non-Small Cell
Lung
Cancer > -4.00 -> -4:00 > -4.00
A549/ATCC >~.po >..4.00
EKVX > -4.00 > -4.00 > -4.00
2 0 HOP-18 > -4.00 > -4.00 > ..4.0o
HOP-62 > -4.00 _ > -4.00 > -4.00
HOP-92 -7.62 -7.02 -6.28
NCI-H226 > -4.00 > -4.00 > .,4.00
NCI-H23 -4.21 > -4.00 > -4.00
2 5 NCI-H322M -6.88 > -4.00 > -4.00
NCI-H460 -4.66 -4.34 r4.02
NCI-H522 -7.44 -6.78
- LXFL 529
- Small Cell Lung
Cancer
3 0 DMS 114 > -4.00 > -4.00 > -4.00
_ DMS Z73 > -4.00 > i.00 > -4.00
CA 02272275 1999-OS-18
WO 98I23269 - l PCT/LTS96/19118
136
Psael/Cel1 Line Loglo GI5081o TGI I-81o I-C50
Colon Cancer
COLO 205 -5.30 > -4.00 > -4.00
DLD-1 > -4.00 > -4.00 > -4.00
HCC-2998 -4.31 >-4.00 >-4.00
HCT-116 > i.00 > -4.00
HCT-15 -4.21 > -4.00 > -4.00 _
HT29 > -4.00 > -4.00 > -4.00
KM 12 > -4.00 > -4.00 > -4.00
._. KM20L2 -6.53 > -4.00 > -4.0Q
-.
SW-620 > -4.00 > -4.00
CNS Cancer
SF-268 ~ ,_ ..., > i.00 >-4.00 > i.00
SF-295 > -4.00 > -4.00 > -4.00
SF-539 > -4.00 > -4.00 > -4. 00 -.
SNB-19 _ > -4.00 > -4.00 > -4.00
SNB-75 -4.18 > -4.00 > i.00
SNB-? 8 > -4.00 > -4.00 > -4.00
U251 > -4.00 f > -4.00
XF 498 > -4.00
2 0 Melanoma -
LOX IMVI >-4.00 >i.00 >-4.00
MALME-3M > -4.00 > -4.00 > -4.00
M 14 > -4.00 > -4.00 > -4.00
M 19-MEL > -4.00 > -4.00 > -4.00
2 5 SK-MEL-2 > -4.00 > -4.00 > -4.00
SK-MEL-28 > -4.00 > -4.00 > -4.00
SK-MEL-5 > -4.00 > -4.00 > -4.00 - _
UACC-257 -6.73 -6.39 -6.04
- UACC-62 -7.24 > -4.00
- CA 02272275
1999-OS-18
WO 98/23269 PCT/US96/19118
137
_ _ P~el/Cell Line I-81o GI50I-a810 I-o8lo
TGI I-C50
Ovarian Cancer
IGROV 1 -0.83 > -4.00
OVCAR-3 -6.67 > -4.00
OV CAR-4 > -4.00 > -4.00 > -4.00
OVCAR-5 -6.35 > -4.00
OV CAR-8 > -4.00 > -4.00 > -4.00
SK-OV-3 -4.39 > -4.00 > -i.00
Renal Cancer
786-0 > -4.00 > -4.00 > -4.00
1 ~ A498 _. - -
ACHN > -4.00 > -4.00 > -4.00
CAKI-1 _ -7.21 fi.44 > -4.00
RXF-393 < -8:00 > -4.00
RFX-631 -4.02 > -4.00 > -4.00 -
SN 12C > -4.00 > i.00 > -4.00
TK-10 -6.86 > -4.00
UO-31 > -4.00 > -4.00 > -4.00
MG MID -4.75 -4.21 -4.08
Delta 3.25 2.81 2.21
2 0 Range 4.00 3.02 2.28
CA 02272275 1999-OS-18
WO 98/23269 ~ PCT/US96/19118
138
NSC: G46270=Z/1
eo>~c
s
PaneI/Cell Line Loglo GI50Loglo TGI Loglo LC50
Leukemia
CCRF-CEM > ~.~ _ > ~.~ > i.00
HL-60(TB) > -4.00 > -4.00 ~ i.00
K-562 > -4.00 > -4.00
MOLT-4 > -4.00 > -4.00 > -4.00
RPMI-8226 > -4.00 > -4.00 > -4.00
SR > -4.00 > -4.00 > -4.00
Non-Small Cell Lung :-
_ Cancer > -4.00 > -4.00 > i.00
A549/ATCC > -4.00 > -4.00 > -4.00
EKVX
2 0 HOP-I 8 > -4.00 > -4.00
HOP-62 > -4.00 > -4.00 > ~. ~ _
HOP-92 -7.88 - -7.44 -7.00
NCI-H226 > -4.00 > -4.00 > i.00
NCI-H23 -5.28 -4.60 > -4.00
2 5 NCI-H322M -7.80 > 4.00 > -4.00
NCI-H460 < -8.00 < -8.00 -7.96
LXFL 529
Small Cell Lung Cancer
DMS 114 > -4.00 > -4.00 > -4.00
3 0 DMS 273
CA 02272275
1999-OS-18
WO 98I23269 PCT/L1S96/1911$
- - __ 139
Panel/Cell Line Loglo GI50 Loglo Loglo LC50
TGI
Colon Cancer
COLO 205 -6.81 -6.34 -S.66
DLD-I > -4.00 > -4.00 > -4.00
HCC-2998 -5.74 -4.69 > -4.00
HCT-116 -7.82 >-4a00 >-4.00
HCT-15 > -4.00 > -4.00 > -4.00
HT29 > -4.00 > -4.00
KM 12 > -4.00 > -4. > -4.00
00
KM20L2 -6. 84 > -4.00 > -4.00 -
SW-620 -7.17 > -4.00 > i.00
CNS Cancer
SF-268 . - > ..4.00 > -4.00 > i.00
SF-295 -4.43 > -4.00 > -4.00
SF-539 >-4.00 >-4.00 >-4,00
SNB-19 -4.47 >-4.00 >-4.00
SNB-75 -4.43 > -4.00 > -4.00 -
SNB-78 ' -4.44 > -4.00 > -4.00
- U251 -7. 73 > -4.00 > -4.00
XF 498 -7.52 > -4.00
2 0 Melanoma
_ - LOX IM > -4.00 > -4.00 > -4.00
V I
MALME-3M >-4.00 >-4.00 >-4.00
M 14 > i.00 > -4.00 > -4.00
M 19-MEL > -4.00 > -4.00 > -4.00
2-5 SK-MEL-2 > -4.00 > -4.00 > -4.00
SK-MEL-28 >..4.00 >-4.00 >-4.00
SK-MEL-5 > -4.00 > -4.00 > -4.00
' UACC-257 -7.87 --w -?.48-7.09
UACC-62 > -4.00 -7.91
- CA 02272275
1999-OS-18
WO 98l23269 PCT/US96/19118
140
_ P~el/Cell Line I-81o GI50 81o TGI Loglo LC50
Ovarian . _
Cancer
IGROV 1 < -8.00 -7.20 -5.31
OVCAR-3 <-8.00 -7.28 >-4.00
OV CAR-4 > -4.00 > 4.00 > -4.00
OV CAR-5 -7.52 -7.01 > -4. 00
-
OVCAR-8 -4.10 >-4.00 >-4.00
SK-OV-3 -5.38 -4.27. >-4.00
Renal Cancer
7 86-0 > -4.00 > -4.00 > -4.00 -
A498 < -8.00 < -8.00 -7.46
ACHN - > -4.00 > -4.00 > -4.00
CAKI- I < -8.00 < -8.00 < -8.00
RXF-393 > -4.00 > -4.00 > ~. 00
RXF-631 < -8.00 -7. 80
SN 12C > -4.00 > -4.00 > -4.00
TK-10 -7.84 -6.96 >-4.00
UO-31 > -4.00 > -4:00 > -4.00
MG MID -5.35 -4.77 -4.37
Delta 2.65 3.23 3.63
2 0 Range 4.00 4.00 4.00
CA 02272275 1999-OS-18
WO 98/23269 ~ PCT/US96/19118
141
NSC: 666710-S/0-1/9
~ _
O
HO 8 S
Paael/Cell Line LoglQ GI50Loglo TGI Loglo LC50
~~emia _--_.
CCRF-CEM -4.55 > -4.00 > -4.00
HL-60(TB) -4.83 -4.43 -4.04
w --15 K-562 ~ -6.36 > -4.00 > -4.00
MOLT-4 -4.83 -4.14 > -4.00
- RPMI-8226 -4.93 -4.17 > -4.00
- - ._.._ . SR -4.96 -4.49 > -4.00
-4.02
Non-Small Cell Lung .
2 0 - Cancer -5.13 -4.60 -4.18
A549/ATCC -4.87 -4.57 -4.27
EKVX -4.87 -4.53 -4.18
HOP-62 -4:98 -6.50 -6.02
NCI-H226 -4.85 -4.44 -4.04
2 5 NCI-H23 -4.97 -4.48 > -4.00
NCI-H460 -4.85 -4.49 -4.10
NCI-H522
CA 02272275 1999-OS-18
WO 98I23269 PCT/US96119118
142
Panel/Cell Line h810 GI50 L810 TGI I-n810
L~0
Colon Cancer
COLO 205 -6.07 ---4.56 -4.21
HCC-2998 -4.39 > -4.00
HCT-116 -4.33 >-4.00
HCT-15 -4.61 -4.18 >-4.00
Ii'I'29 -4.51 -4.87 _,
SW-620 -4.34 >-4.00
CNS Cancer
SF-268 -4.82 -4.49 -4.17
~ SF-295 -4.74 -4.45 -4.21
SF-539 -4.72 -4.46 -4.20
SNB-19 -4.79 -4.44 -4.09
SNB-75 -5.21 -4:72 -4.36
U251 -594 -4.68 -4.33
Melanoma
- LOX IMVI -4.85 -4.11 >-4.00
MALME-3M -4.64 -4.35 -4.64
M 14 i.84 -4:53 -4.23
SK-MEL-2 -4.69 -4.32 > -4.00
2 0 SK-MEL-28 _ -4.78 -4.50 -4.22
SK-MEL-S -4.94 -4.6Z -4.31
UACC-257 -7.45
UACC-62 < -8.00 -4.83 -4.37
Ovarian Cancer
2 5 IGROV 1 -7.77 -6.68
OVCAR-4 -4.79 -4.43 -4.08
OVCAR-5 -7.19 __.:-6.44 -4.40
OVCAR-8 -4.59 >-4.00 >-4.00
SK-OV-3 ' -5.12 -4.67 -4.32
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PanelJCel1 Line Loglo GI50Loglo -TGILoglo LC50
Renal Caacer
- 786-0 . -4.73 -4.47 - -4.21
ACHN -4.80 -4.51 -4.23
CAKI-1 -7.84 ~. ~
RXF-393 -6.74 -4.80 -4.37
SN12C -4.66 -4.26 >-4.00
TK-10 -6.96 -4.48
UO-31 -4.83 -4.56 -4.25
Prostate Cancer
PC-3 -4.72 -4.29 > -4.00
DU-145 -4.79 -4.48 -4.18 ..
Breast Cancer
MCF7 -7.35 -4.28
MCF7/ADR-RES -4.48 >-4.00 >-4.00
MDA-MB-231/ATCC -4.64 -4.21 >-4.00
HS 578T -4.75 -4.27 > -4.00
MDA-N . -4.87 -4.54 -4.21
BT-549 -4.85 -4.39 > -4.00
T-47D -5.64 -4.57 > -4.00
2 0 MG MID -5.37 -4.55 -4.17
Delta w 2.63 2.32 1.82
Range 3.52 2.88 2.02
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NSC: 666711-T/0-1/10
-
"o i I I I I I
~o .
Panel/Cell Line Loglp GI50Loglp TGI Loglp LC50
~~e~a _
CCRF-CEM -5.16 > -4.00 > -4.00
HL-60(TB)
K-S62 -6.71 > -4.00 > -4.00
MOLT-4 i.74 -4.16 > -4.00
_ RPMI-8226 =5.26 -4. 36 > -4.00
SR -4.87 -4.4S -4.04
Non-Small Cell Lung
Cancer -4. 83 > -4.00 > -4.00
2 0 A549/ATCC -4.64 > -4.00 > -4.00
EKVX -4.46 > -4.00 > -4.00
HOP-62 -7.88 -7.35 -b.68
NCI-H226 -4.55 > -4.00 > -4.00
NCI-H23 fi.96 > -4.00 > -4.00
2 5 NCI-H460 -4.31 > -4.00 > -4.00
NCI-H522
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Panel/Cel1 Line Loglo GI50Loglo TGI Loglo LC50
Colon Cancer
COLD 205 i.28 -4.11 > -4.00
HCC-2998 ..6.72 -4.42 > -4.00
HCT-116 -7.24 >-4.00 >-4.00
" 5 HCT-15 -6.30 > -4.00 > -4.00
KM 12 -6.64 > -4.00 > -4.00
- SW-620 > -4.00 > -4.00
- CNS Cancer
- . . SF - _4.57 > ~.~ > i.00
_ 268
SF-295 -4.58 > -4.00 > -4,00 -
SF-539 -4.32 > -4.00 > -4.00.
-
SNB-19 > -4.00 > ..4.00 > -4.00
SNB-75 -4. S4 > -4.00 > -4.00
U251 -4.62 > -4.00 > ~.~
Melanoma
LOX IMVI -5.15 >-4.00 >-4.00
MALME-3M -6.63 >-4.00 >-4.00
M 14 ~ -4.51 > -4.00 > -4.00
SK-MEL-2 > -4.00 > -4.00 > -4.00
_. _ _ ~ 0 SK-MEL-28 -4.14 > -4.00 > -4.00
SK-MEL-5 -4.82 -4.47 -4.33
_ - _. UACC-257 -7.32 -6.58 -4.04
UACC-62 _ -7.54 -4.72 -4.23 -
Ovarian Cancer -
2 5 IGROV 1 -7.79 -6.68 > -4.00
OVCAR-4 > -4.00 > -4.00 > -4:00
OVCAR-5 -7.11 -6.43 >-4.00
OV CAR-8 -4.63 > -4.00 > -4.00
_ SK-OV-3 -5.78 -4.57 >-4.00
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Panel/Cell Line Loglo GI50Loglo TGI Loglo
LC50
Renal Cancer
786-0 > -4.00 > -4.00 > -4.00
ACHN -4.45 > -4:00 > -4.00
CAKI-1 -7.93 > -4.00
RXF-393 -7.34 > -4.00 > -4.00
SN 12C > -4.00 > i.00 > -4.00
TK-10 -7.41 -6.53 > -4.00
UO-31 -4.13 > -4.00 > -4.00-
Prostate Cancer
PC-3 -4.54 > -4.00 > ~.~
DU-145 -4.07 > -4.00 > -4.00
Breast Cancer
MCF7 -7.88 -7.2i > -4.00
MCF7/ADR-RES > -4.00 > -4.00 > -4.00
i 5 MDA-MB-231 /ATCC > -4.00 > ~.DO > -4.00
HS 578T -4.64 >-4.00 >-4.00
MDA-N -4.41 > -4.00 > -4.00
BT-549 -4.56 -4.12 - > -4.00
T-47D -6.24 > -4.00 > -4.00
2 0 MG MID -5.3 i -4.39 4.06
Delta 2.62 2.96 2.62
Range 3.95 3.35 2.68
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NSC: 665150.W/0-1/15
~ ( _
s ' s I ~o
Panel/Cell Line Logo GI50 Loglo TGI Loglo LC50
Leukemia
CCRF-CEM -4:S9 -4.20 >-4.00
HL-60(TB) . _ .~_. -4.36 -4.15 >-4.00
_:. ..
K-562 -4.99 -4.12 > -4.00
MOLT-4 -4.54 -4.08 >-4.00
RPMI-8226 - -4.70 -4.17 > -4.00
SR -4.52 -- -4.13 > -4.00
Non-Small Cell Lung -
Cancer -4. 88 i.30 > -4.00
A549/ATCC -4.05 > -4.00 > -4.00
2 0 EKVX -5.76 -4.49 > -4.00
HOP-62 -4.29 > -4.00
HOP-92 -6.96 -6.43 -5.75
NCI-H226 -4. 80 -4.14 > -4.00
NCI-H23 -4.89 -4.56 -4.23
2 5 NCI-H322M -6.31 -5.71 -5.24
NCI-H460 -4.29 > -4.00 > -4.00
NCI-H522
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PanellCell Line Loglo GI50Loglo TGI Loglo LC50
Colon Cancer
COLO 205 -5.26 -4.72 -4.25
HCC-2998 -4. 83 -4.18 > -4.00
HCT-116 -6.39 -4.79 -4.39
HCT-15 -4.29 >-4.00 >-4.00
HT29 -5.79 -4.89 > -4.00
KM 12 -4.11 > -4.00 > -4.00
.- SW-620 -5. Z5 > -4.00 > -4.00
CNS Cancer
. ~ SF-268 -4.57 i.07 >-4.00
SF-295 -4.02 > -4.00 > -4.00
SF-S39 -4.62 -4.28 > -4.00
SNB-19 -4.47 > -4.00 > -4.00
SNB-75 -4.33 - > i.00 > -4.00
U251 -4.66 -4.08 > -4.00
M~~~
MALME-3M -4.10 >-4.00 >-4.00
M 14 -4.42 > -4.00 > -4.00
SK-MEL-2 -4.60 -4.08 > -4.00
2 0 SK-MEL-28 -4.55 -4.09 > -4.00
SK-MEL-5 -4.97 -4.59 -4.22
UACC-257 -7.90 i.82 i.36
UACC-62 -7.17 -6.49 -5.57
Ovarian Cancer
2 5 IGROV 1 -6.16 -5.50 -4.64
OVCAR-3 -6.22 -5.08 -4.11
OVCAR-4 -5.27 > -4.00 > i.00
OVCAR-5 -7.83 -6.73
- OVCAR-8 -4.84 -4.41 >-4.00
30 SK-OV-3 -4.69 -4.14 >-4.00
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Panel/Gell _ Line 81o GI50 81o TGI Logio LC50
Renal Cancer
786-0 -4.64 > -4.00 > 4.00 ~ . _. -..
A498 i.68 -6.39 -6.10
ACHN -4. 30 > -4.00 > -4.00
CAKI-1 -5.46 -S.02 > -4.00
RXF-393 -4.61 ~.~ .. > ~.~
SN12C -4.S3 >-4.00 >-4.00
TK-10 -6.65 -b.23 >-4.00
- UO-31 -4.28 > -4.00 > -4.00
Prostate Cancer
PC-3 -4. 32 > -4.00 > -4.00
DU-145 -5.12 -4.49 >-4.00
Breast Cancer -
MCF7 -7.11 -4.94 ' -4.01
MCF7/ADR-RES -4.77 -4.08 >-4.00
MDA-MB-231/ATCC -4.66 -4.19 >-4.00
HS 578T -4.61 -4.16 > -4.00
MDA-MB-435 -4.60 -4.23 > -4.00
MDA-N -4.44 > -4.00 > -.4.00
2 0 T-47D _. -4.41 > -4:00 > -4.00
MG MID -5.09 -- -- -4.5I -4.19
Delta 2.81 2.32 2.17
Range 3.87 2.82 2.36
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Example 50
In Vivo Antitumor Activity
The following experiments were carried out at Purdue
University athymic mouse laboratory. Athymic mice-were
purchased from and maintained at the athymic mouse
laboratory under pathogenic conditions. Tumor cells (TBE)
were obtained from normal human bronchial epithelial cells
transfected~with p2~smic H1 carrying v-Harvey-ras (H-ras)
oncogene via protoplast fusion (G. H. Yoakum, J.F. Lechner,
E.W. Gabrielson, B.E. Korba, L. Malan-Shibley, J.C. Willey,
M.G. Valerio, A.M. Shamsuddin, B.F. Trump and C.C. Harris,
Science 227, 1174, 1985.. T.C.K. Chan, C.-j. Chang, N.M.
Koonchanok and R.L. Geahlen, Biochem. Biophys. Res.
Commun., 193, 1152, 1993. The TBE cells were implanted
subcutaneously on day 0 of tha experiment. Each test group
consists of five mice. The compounds tested were dissolved
in a saline solution or a saline solution containing 5% of
dimethylsulfoxide and 10% cremphor EL (Sigma Cheltiical'
Company, St. Louis, MO), and were administered
intraperitoneously.starting on day 2. The solution was
administered every four days for a total of four
treatments. Tumor volumes were measured every week. The -
antitumor activity was measured as-% of growth inhibition,
which is defined by the percentage~of the median tumor
volume reduction per week of the treated mice divided by
the median tumor volume per-week of the controlled mice
(Tables 9 and 10).
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Table 9.---- Ant,itumor Efficacy of 5-Aminomethyl-a-
terthiopheae Tartarate-Hydroxypropyl-~-cyclodeztrin Complex
(N8C Code ~6606~1)
Tumor cells: ras-transformed human bronchial epithelial
cells (2.5 x 106 cells/mouse)
Formulation: saline solution
1. control
Number of Tumor Volume Slope Tumor Growth
week Animals with (mm3) (mm3/week)
Inhibition Tumor l%)
1 1 1.3
2 1 1.3
3 3 14.7 4.41
4 3 16.0 4.54
5 3 19.3 4.40
6 3 24.3 4.41
7 3 34:7 4.92
8 3 34.3 4.80
- 20
2. 12 mg/~cg
Number of Tumor Volume
Slope
Tumor-Growth
_ (mm3) (mm3/week)
Week Animals with
Inhibition Tumor (%)
1 0 0:0-
2 0 0.0
- 3- 1 - 1.2 0.36 92
4 2 4.8 1.08 76
5 2 4.8 1.13 74
6 2 4.8 1.03 77
_ 7 4 8.2 1.18 76
8 5- 24.0 2.34 51
* Tumor Growth Inhibition = Slope ntrol-1,- Slopeeated) x100
(%) (co (tr
Slope (control)
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Table i0. Antitumor Efficacy of 5-HydroBpmethyl-5~~-
.formyl-a-terth~o~hene (NBC Code ~6~t7~55) -
Tumor cells: ras-transformed human bronchial epithelial
cells (4.0 x 106 cells/mouse)
Formulation: 5% DMSO and 10% cremphor in saline solution
i. control
Number of Tumor Volume Slope Tumor Growth
Week Animals with (mm3) (mm3/week)
Inhibition Tumor (%)
1 2 2.4
_ 1.5 3 6.2
2 4 10.8
3 4 14.6 4.97
4 4 15.8 4.37
5 4 15.0 3.45
6 4 16.6 2.91
8.6 5 23.6 2.63 -- -
2. 25 mg/kg
Number of Tumor Volume Slope Tumor Growth
Week Animals with (mm3) (mm3/week)
Inhibition Tumor (%)
1 0 0.0
1.5 1 1.4
2 ~ 3 5 , 2 _.. -.-_
3 4 5.8 2.16 57
- 4 4 5.6 1.75 60
5 4 5.4 1.34 61
6 5 - 10.6 1.60 45
8.6 5 18.8 2.07 21
* Tumor Growth Inhibition ($) = Slope (control) - Slope (treated) x 100
3 5 Slope (control)
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Fxample 51
Human tumor cells of various origins (kidney, skin,
ovary and lung cell lines) were implanted into athymic mice
and injected intraperitoneally with polythiophene compounds -
of the present claims. The tumor cells were-implanted at
various sites: subcutaneous, intraperitoneal and subrenal.
At predetermined times after tumor implantation,
l0 polythiophene compositions were administered
intraperitoneally to the test group mice according to the
drug schedule indicated by code in table il. The first
numbe-r of the cods indicates the interval between
treatments; the secorid number indicates the Total number of
times the drug was administered and the superscript letter
indicates the first day of treatment. Thus the designation ---
Q4d x 3d indicates the treatments were initiated 15 days
after implantation and a total of three treatments were
administered at intervals of four days. Tumor growth
inhibition was determined by comparing the wolume/weight of
---~_ _
the tumors o~ the treated mice in the test group with that
of-the tumors in the control group mice. The data are
presented as the percent of treated tumor volume/weight to
control tumor volume/weight (T/C %) with the day the
measurement was taken indicated in parenthesis.
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- - ..._ 154
Table lI. Evaluation of IR iw~o Mtitumor Efficacy of Polythiopha~es
Cod Implant' _ Drug' Drug T/C 96t
NSC,t Tumor (Origin) Site Dosage(mg/lcg) Schedule (Day)
660641 A-~98(kidney) SC 173- Q4D x 3' 46(Z9)
CAKI-1(kidney) SC 157 QD x5' 44(36)
UACC-257(skin) SC 17S Q7D x3' 53(33)
- ~ -' TBE(lung) SC 155 Q4D x4' 24(49)
P-388(Iymphocyte)IP 225 QDxS~ Inactive
647455 TBE (lung) SC ZS Q4Dx 4~ 39(35)
637388 CAKI-1 (kidney) SR 50 Q4Dx3' 16(13)
658878 TBE pang) SC 40 Q4D x4 5(70)
- 666710 A~98(kidney) SC 60 Q4Dx4j 45(38)
3 0 a. 660641: hydroxypmpyl-B-cyclodextrin inclusion complex of 2-aminohydroxy-
a-
~~ph~e tartarated (2:1 molar ratio; FW:3428) (Estimated 5096 lethal
dosage: 1550 mg/kg.)
647455: 2-formyl-5"~ydroxymethyl-a-to thiophene
637388: Z-hydroxymethyl-or-terthiophene
3 5 658878: 2,4"-dihydroxymethyl-oc-terthiopheae
666710: 2-hydroxymethyl-a-terthiophene ~S-D-glucoside
b. SC: subcutaneous; IP:intraperitoneal; SR: subreaal
c. Drug was administered intraperitoneally.
d-j. First day of drug treamient, d: Day 15; e: Day 12; f:Day 3; g: Day 1; h:
Day 2; i: Day
40 14; j: DaylO.
k. T/C: tumor volume or weight of treated miceltumor volume or weight of
control
(untreated) mice. The number shown in the parenthesis is the day when the
tumor
volume or weight is measured.
