Targeted Detection of Cyclooxygenase-1 in Ovarian Cancer

Article abstract of DOI:10.1021/acsmedchemlett.9b00280

Overexpression of cyclooxygenase-1 (COX-1) is associated with the initiation and progression of ovarian cancer, and targeted imaging of COX-1 is a promising strategy for early detection of this disease. We report the discovery of N-[(5-carboxy-X-rhodaminy

Full text of DOI:10.1021/acsmedchemlett.9b00280

Letter
pubs.acs.org/acsmedchemlett
Cite This: ACS Med. Chem. Lett. XXXX, XXX, XXX−XXX
Targeted Detection of Cyclooxygenase‑1 in Ovarian Cancer
Paola Malerba,†,‡ Brenda C. Crews,† Kebreab Ghebreselasie,† Cristina K. Daniel,† Elma Jashim,†
Ansari M. Aleem,† Redoan A. Salam,†,∥ Lawrence J. Marnett,† and Md. Jashim Uddin*,†
†A. B. Hancock, Jr. Memorial Laboratory for Cancer Research, Departments of Biochemistry, Chemistry, and Pharmacology,
Vanderbilt Institute of Chemical Biology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville,
Tennessee 37232 United States
‡Department of Pharmacy and Pharmaceutical Sciences, University of Bari “A. Moro”, Via Orabona 4, 70125 Bari, Italy
∥Department of Biology, Washington University in St. Louis, St. Louis, Missouri 63130 United States
S
* Supporting Information
ABSTRACT: Overexpression of cyclooxygenase-1 (COX-1) is associated
with the initiation and progression of ovarian cancer, and targeted imaging
of COX-1 is a promising strategy for early detection of this disease. We
report the discovery of N-[(5-carboxy-X-rhodaminyl)but-4-yl]-3-(1-(4-
methoxyphenyl)-5-(p-tolyl)-1H-pyrazol-3-yl)propenamide (CMP) as the
first COX-1-targeted optical agent for imaging of ovarian cancer. CMP
exhibits light emission at 604 nm (λmax), thereby minimizing tissue
autofluorescence interference. In both purified enzyme and COX-1-
expressing human ovarian adenocarcinoma (OVCAR-3) cells, CMP inhibits
COX-1 at low nanomolar potencies (IC50 = 94 and 44 nM, respectively). CMP’s selective binding to COX-1 in OVCAR-3 cells
was visualized microscopically as intense intracellular fluorescence. In vivo optical imaging of xenografts in athymic nude mice
revealed COX-1-dependent accumulation of CMP in COX-1-expressing mouse ovarian surface epithelial carcinoma (ID8-
NGL) and OVCAR-3 cells. These results establish proof-of-principle for the feasibility of targeting COX-1 in the development
of new imaging and therapeutic strategies for ovarian cancer.
KEYWORDS: Optical imaging, cyclooxygenase-1, ovarian cancer, COX-1-selective inhibitor, CMP probe
diffuses from ovarian cancers and cooperates with vascular
endothelial growth factor to induce release of Fas ligand from
vascular endothelial cells. Fas ligand acts on CD8+ T cells to
induce apoptosis, thereby preventing their cytotoxic action on
ovarian cancer cells. Thus, the combination of a vascular
endothelial growth factor receptor kinase inhibitor with aspirin
(a nonselective COX inhibitor) dramatically inhibited the
growth of ovarian cancers in vivo.9 Recently, a large
consortium that pooled analyses of 12 studies demonstrated
that daily low-dose aspirin confers a 20 to 34% reduction in the
risk of developing ovarian cancer.10 Furthermore, large
observational studies in patients show that regular use of
aspirin, even at low doses, reduces the long-term risk of distant
metastases, particularly in the case of adenocarcinomas.11
These results support the functional significance of COX-1 in
ovarian cancer and the potential value of COX-1-targeted
imaging and therapeutic agents in the management of this
neoplastic disease.
he cyclooxygenase enzymes (COX-1 and COX-2)
Tcatalyze the initial, rate-limiting step in the conversion
of arachidonic acid to prostaglandins (PGs) and thromboxane.
Expression patterns of the two enzyme isoforms support the
hypothesis that COX-1 plays a predominantly homeostatic
role, whereas COX-2 is a key modulator of inflammation,
cellular proliferation, and tumorigenesis.1,2 In fact, COX-2
overexpression is a well-documented phenomenon in many
forms of cancer.3,4 An exception to these generalities, however,
is seen in the case of ovarian cancer, in which COX-1 is
frequently overexpressed and believed to contribute signifi-
cantly to disease initiation and progression.5 For example, Li
and co-workers reported COX-1 expression in seven out of
nine ovarian carcinoma cell lines, whereas COX-2 expression
was not detectable.6 In addition, elevated COX-1 but not
COX-2 expression was detected in genetically engineered
murine ovarian cancer cell lines that lack p53 and overexpress
either myc and K-ras or myc and akt. Tumors derived by
orthotopic implantation of these cell lines in nude mice also
expressed high levels of COX-1, and growth of one of them in
vivo was suppressed by the COX-1-selective inhibitor SC-560,
but not the COX-2-selective inhibitor celecoxib.7 Studies
suggest that PGs derived from COX-1-dependent oxygenation
of arachidonic acid play an important role in multiple events
associated with growth, survival, and metastasis of ovarian
cancer.8 Motz et al. reported that COX-1-derived PGE2
Clinical diagnosis of ovarian cancer in its early stages is a
major unmet medical need. Existing strategies for diagnosis of
Special Issue: Medicinal Chemistry: From Targets to Therapies
Received: June 21, 2019
Accepted: July 24, 2019
Published: July 24, 2019
© XXXX American Chemical Society
A
DOI: 10.1021/acsmedchemlett.9b00280
ACS Med. Chem. Lett. XXXX, XXX, XXX−XXX

ACS Medicinal Chemistry Letters
Letter
Figure 1. (a) Structures of SC-560, celecoxib, and fluorocoxib A, and (b) chemical synthesis of CMP.
ovarian cancer include ultrasound, computed tomography,
magnetic resonance imaging, position emission tomography
(PET), and laparoscopy. Among these methods, only PET
employs contrast agents, such as [18F]fluorodeoxyglucose
([18F]FDG), for visualization. In fact, [18F]FDG-PET has
been evaluated for the diagnosis of ovarian cancer in a number
of clinical studies,12 but it has not demonstrated adequate
specificity or sensitivity in these settings.13,14 Other imaging
agents that have been developed so far have shown some
promise in animal models of ovarian cancer, but none has
progressed into clinical trials. Our laboratory has made major
contributions to the development of targeted imaging15−19 and
therapeutic agents20,21 for cancer. We discovered the first
COX-2-targeted optical imaging agent (fluorocoxib A)15 and
cancer chemotherapeutic agent (chemocoxib A),20 and we
have also discovered COX-2-targeted PET and single-photon
emission computed tomography agents for imaging of cancer
and inflammation.22,23 Herein, we report the first COX-1-
targeted optical imaging agent N-[(5-carboxy-X-rhodaminyl)-
but-4-yl]-3-(1-(4-methoxyphenyl)-5-(p-tolyl)-1H-pyrazol-3-
yl)propenamide (CMP) and demonstrate its feasibility in the
COX-1-dependent in vivo visualization of human ovarian
tumor xenografts.
The discovery of the COX-2-targeted imaging agent
fluorocoxib A (Figure 1a) was accomplished through the
conjugation of a fluorescent moiety to the carboxyl group of
the nonselective COX inhibitor indomethacin via a hydro-
carbon tether. The resulting compound exhibited COX-2-
selective inhibitory activity, and we found that we could, in
fact, generate a wide variety of fluorescent inhibitors in this
fashion.19 The X-ray crystal structures of fluorescent
indomethacin−dansyl conjugates in complex with COX-2
revealed that the indomethacin moiety adopts a binding pose
in the enzyme’s active site very similar to that of the parent
compound and that the tether extends through a constriction
at the opening of the active site, placing the dansyl moiety in
the lobby, an alcove within the protein’s membrane binding
domain.24 The relatively spacious lobby allows it to
accommodate conjugates as large as 5-carboxy-X-rhodamine
as is found in fluorocoxib A. We hypothesize that we could
apply the same general strategy to develop a COX-1-targeted
optical imaging agent, starting with the COX-1-selective
inhibitor SC-560 as a scaffold. An X-ray crystal structure of
mefozolac, a COX-1-selective diarylheterocycle similar to SC-
560, in complex with COX-1 revealed a binding orientation
that placed the inhibitor’s five-membered heterocyclic ring
close to the constriction, suggesting that attaching the tether to
this ring would most likely lead to success.25
To test this hypothesis, we synthesized CMP using a
multistep reaction scheme (Figure 1b; full compound names
are provided in the Supporting Information). Briefly, the
Claisen condensation of compound 1 with succinic anhydride
in the presence of lithium diisopropylamide proceeded
smoothly to afford compound 2. Condensation of 2 with (4-
methoxyphenyl)hydrazine hydrochloride afforded compound
3. After the 1,5-regioisomer was separated from the minor 1,3-
regioisomer by flash chromatography, 3 was then reacted with
tert-butyl (4-aminobutyl)carbamate in the presence of ethyl-1-
[3-(dimethylamino)propyl]-3-ethylcarbodiamide to give a
carbamate intermediate, which was then treated with hydrogen
chloride gas to afford compound 4. 5-Carboxy-X-rhodamine-
acid was activated using N,N,N′,N′-tetramethyl-O-(N-
succinimidyl)uronium tetrafluoroborate in the presence of
triethylamine and coupled with 4 to yield the target fluorescent
conjugate CMP (compound 5). CMP emitted bright red-
shifted fluorescence with excitation and emission maxima of λex
B
DOI: 10.1021/acsmedchemlett.9b00280
ACS Med. Chem. Lett. XXXX, XXX, XXX−XXX

ACS Medicinal Chemistry Letters
Letter
Figure 2. (a) Photophysical properties of CMP; (b) purified COX-1 and COX-2 inhibition profiles of CMP; (c) cellular COX-1 inhibition profile
of CMP in OVCAR-3 cells; (d) immunoblot showing the expression of COX-1 and COX-2 in the ID8-NGL and OVCAR-3 cell lines; (e) optical
imaging of COX-1 in OVCAR-3 cells; (f) imaging of COX-1 in OVCAR-3 cells pretreated with indomethacin (5 μM).
= 580 nm and λem = 606 nm, respectively, and a quantum yield
of 0.95 at 604 nm (Figure 2a). The compound’s molar
extinction coefficient was 35/mM/cm at 580 nm. These
optical properties are desirable for an in vivo optical imaging
agent, as they help to minimize the problem of autofluor-
escence interference from live tissues.
The ability of CMP to inhibit purified COX-1 or COX-2 was
evaluated in a thin layer chromatography assay using
[1-14C]arachidonic acid as substrate.15 Briefly, purified ovine
COX-1 or murine COX-2 was preincubated with vehicle or
CMP at various concentrations for 20 min followed by
substrate addition and incubation for an additional 30 s.
C
DOI: 10.1021/acsmedchemlett.9b00280
ACS Med. Chem. Lett. XXXX, XXX, XXX−XXX

ACS Medicinal Chemistry Letters
Letter
Termination solution was added, and the products were
extracted, spotted, developed, and scanned for [1-14C]-
arachidonic acid remaining. CMP displayed selective and
potent COX-1 inhibitory activity (COX-1 IC50 = 94 nM;
COX-2 IC50 > 1000 nM) (Figure 2b). Inhibition of COX-1 in
intact cells by CMP was also evaluated using a previously
described method.15 COX-1-expressing OVCAR-3 cells were
treated with vehicle or CMP at different concentrations for 30
min followed by addition of [1-14C]arachidonic acid and
incubation for 30 min more. Aliquots of the culture medium
were transferred to termination solution, extracted, spotted,
developed, and scanned for [1-14C]arachidonic acid remaining.
The IC50 value for inhibition of human COX-1 by CMP in
OVCAR 3 cells was 44 nM (Figure 2c).
Using commercially available specific antibodies, we tested
the levels of COX-1 and COX-2 in human OVCAR-3 ovarian
cancer cells and ID8-NGL murine ovarian surface epithelial
cancer cell lines by immmunoblot analysis. We used purified
COX-1 and COX-2 as standards and β-actin as a loading
control. We detected high levels of COX-1 expression in
OVCAR3 and ID8-NGL cells as compared to minimal COX-2
expressions in these cell lines (Figure 2d). ImageJ software
version 1.52K (National Institutes of Health, USA) was used
to analyze the intensity of obtained bands. The immmunoblot
data suggest these cell lines will be suitable for evaluating and
validating the feasibility of CMP in vitro and in vivo.
We evaluated the ability of CMP to fluorescently label COX-
1 in OVCAR-3 cells in culture. Following pretreatment of the
cells with vehicle or indomethacin for 30 min, the cells were
incubated with CMP for an additional 30 min, washed briefly,
and then cultured in serum-containing medium for 45 min.
The purpose of indomethacin pretreatment was to block the
active site of COX-1, thereby preventing uptake and retention
of CMP. Optical imaging under a fluorescence microscope
revealed that vehicle-pretreated cells were reproducibly labeled
by CMP (Figure 2e), whereas minimal fluorescence was
observed in indomethacin-pretreated cells (Figure 2f). The
results suggest that CMP’s fluorescence labeling in vehicle-
treated cells was COX-1-dependent. Notably, the dispersed
cytoplasmic fluorescence signals denote a diffuse expression of
COX-1 in these cells.
We validated CMP as a COX-1-targeted optical imaging
agent in two different mouse tumor models of ovarian cancer
using a protocol approved by the Vanderbilt Institutional
Animal Care and Use Committee. First, we evaluated the
ability of CMP to detect COX-1 in xenografts derived from
OVACR-3 cells. In each female athymic nude mouse, OVCAR-
3 cells were implanted subcutaneously on the left hind flanks.
The xenografts were allowed to grow to approximately 750−
1000 mm3 to ensure that a vascular bed was established. Mice
were then administered CMP intraperitoneally (i.p., 1 mg/kg
in dimethyl sulfoxide (DMSO)), and at 15 min, 30 min, 1 h, 2
h, 3 h, and 4 h postinjection, they were lightly anesthetized and
placed in an IVIS imaging system. No detectable level of CMP
uptake was observed at any time points before 2 h
postinjection. However, at 3 to 4 h postinjection, we detected
reproducible, high tumor fluorescence as compared to leg
muscle signals (Figure 3a). To further check the biodistribu-
tion, after in vivo imaging, we collected tumor, muscle, kidney,
and liver from OVCAR-3 tumor-bearing animals and imaged
them ex vivo (Figure 3b). The ex vivo quantification showed
high CMP accumulation in the OVCAR-3 tumors (Figure 3c)
compared to the other tissues (n = 4, p < 0.004).
Figure 3. (a) In vivo imaging of COX-1 in an OVACR-3 tumor by
CMP; (b) ex vivo imaging of an OVACR-3 tumor compared with leg
muscle, kidney, and liver; (c) region of interest measurements of light
emission from OVACR-3 tumor, leg muscle, kidney, and liver (CMP,
t = 3.5 h) (RFU, relative fluorescence units).
We evaluated the utility of CMP in COX-1-targeted imaging
of ovarian tumors derived from ID8-NGL cells implanted into
the left hind flanks of nude mice (n = 6 animals) using a
procedure similar to that described above. For imaging, we
administered CMP i.p. (2 mg/kg in DMSO) to mice bearing
full grown tumors (750 to 1000 mm3). The animals were
imaged under light anesthesia at 3 to 4 h postinjection of
CMP. The ID8-NGL tumors exhibited strong fluorescence
signals (Figure 4a and Supplemental Figure S1). After in vivo
imaging, we euthanized the animals and collected the ID8-
D
DOI: 10.1021/acsmedchemlett.9b00280
ACS Med. Chem. Lett. XXXX, XXX, XXX−XXX

ACS Medicinal Chemistry Letters
Letter
604 nm and inhibits COX-1 at low nanomolar potencies in
both purified enzyme preparations and ovarian cancer cell
lines. In vivo optical imaging shows selective uptake of CMP in
COX-1-overexpressing mouse tumors derived from human
ovarian cancer (OVCAR-3) cells and mouse ovarian surface
epithelial carcinoma (ID8-NGL) cells. Uptake of CMP is
COX-1-dependent as confirmed by failure of CMP uptake in
indomethacin-pretreated mouse tumors derived from COX-1-
overexpressing ID8-NGL cells. These data provide support for
the feasibility of targeting COX-1 in pathological tissues. CMP
represents the first COX-1-targeted in vivo optical imaging
agent validated for visualization of tissues, such as the
neoplastic tissues in the ovary, overexpressing the enzyme.
ASSOCIATED CONTENT
* Supporting Information
■
S
The Supporting Information is available free of charge on the
ACS Publications website at DOI: 10.1021/acsmedchem-
lett.9b00280.
Chemical synthesis and spectroscopic characterization of
compounds 2 through 5, methods for animal experi-
ments, and Figure S1 (PDF)
AUTHOR INFORMATION
Corresponding Author
*Phone: 615-484-8674. E-mail: jashim.uddin@vanderbilt.edu.
■
ORCID
Lawrence J. Marnett: 0000-0002-7834-6285
Md. Jashim Uddin: 0000-0003-0020-4866
Author Contributions
P.M. synthesized and characterized all the raw materials and
the final compound CMP; B.C.C. performed animal and cell
imaging experiments; K.G. biochemically evaluated the
compounds; C.K.D. carried out cell-based testing of com-
pounds; E.J. purified a sample of CMP for testing and
deposited CMP for institutional registration; A.M.A. quantified
protein expression by immunoblot analysis, R.A.S. synthesized
a batch of CMP, L.J.M. provided laboratory space and financial
support, and M.J.U. conceived the idea, designed the
experiments, interpreted the data, wrote the manuscript, and
supervised the overall conduct of the research program. All
authors have given approval to the final version of the
manuscript.
Figure 4. (a) In vivo optical imaging of COX-1 in ID8-NGL tumor
(left mouse) and indomethacin-pretreated ID8-NGL tumor (right
mouse); (b) ex vivo imaging of ID8-NGL (left mouse) and
indomethacin-pretreated ID8-NGL (right mouse) tumors compared
with leg muscle; (c) region of interest measurements of light emission
from ID8-NGL tumor and indomethacin-pretreated ID8-NGL tumor
at 3.5 h postinjection of CMP (RFU, relative fluorescence units).
Funding
This program was financially supported by research grants
from the National Institutes of Health CA128323-4, -5 (to
M.J.U.), CA182850-01A1 (to M.J.U.), CA136465 (to L.J.M.),
and CA89450 (to L.J.M.).
NGL tumors and surrounding muscle for ex vivo imaging
(Figure 4b). Based on the measurements of light emission, the
uptake of CMP in the ID8-NGL tumors was ∼8-fold higher
than that in the muscle (Figure 4c). We elucidated the
mechanism of the COX-1-targeted delivery and binding of
CMP in ID8-NGL tumor allografts by administering a high
dose (10 mg/kg in DMSO) of indomethacin i.p. to tumor-
bearing mice 1 h prior to CMP (i.p., 2 mg/kg in DMSO). At 3
to 4 h post-CMP injection, optical imagining revealed
extremely low levels of fluorescence in the tumors, suggesting
effective COX-1 blockade by indomethacin (Figure 4a−c).
These findings support the hypothesis that uptake and
retention of CMP in the tumor was COX-1-dependent.
In conclusion, we describe the discovery of CMP, a COX-1-
targeted optical imaging agent. CMP exhibits light emission at
Notes
The authors declare no competing financial interest.
ACKNOWLEDGMENTS
■
We are grateful to the Vanderbilt Fluorescence Spectroscopy
Core Lab for the use of the PTI photon counting fluorometer,
the Vanderbilt University Institute of Imaging Sciences for the
use of the Xenogen IVIS 200 imaging system, the Vanderbilt
Small Molecule NMR Facility for the use of the Bruker AV-I at
600 MHz instrument, and the Vanderbilt Mass Spectroscopy
Research Center for use of the Quantum Triple Quadrupole
instrument. In addition, we are grateful to Dr. Carol A. Rouzer
E
DOI: 10.1021/acsmedchemlett.9b00280
ACS Med. Chem. Lett. XXXX, XXX, XXX−XXX

ACS Medicinal Chemistry Letters
Letter
(14) Tawakol, A.; Abdelhafez, Y. G.; Osama, A.; Hamada, E.; El
Refaei, S. Diagnostic performance of 18F-FDG PET/contrast-
enhanced CT versus contrast-enhanced CT alone for post-treatment
detection of ovarian malignancy. Nucl. Med. Commun. 2016, 37, 453−
460.
(15) Uddin, M. J.; Crews, B. C.; Blobaum, A. L.; Kingsley, P. J.;
Gorden, D. L.; McIntyre, J. O.; Matrisian, L. M.; Subbaramaiah, K.;
Dannenberg, A. J.; Piston, D. W.; Marnett, L. J. Selective visualization
of cyclooxygenase-2 in inflammation and cancer by targeted
fluorescent imaging agents. Cancer Res. 2010, 70, 3618−3627.
(16) Uddin, M. J.; Werfel, T. A.; Crews, B. C.; Gupta, M. K.;
Kavanaugh, T. E.; Kingsley, P. J.; Boyd, K.; Marnett, L. J.; Duvall, C.
L. Fluorocoxib A loaded nanoparticles enable targeted visualization of
cyclooxygenase-2 in inflammation and cancer. Biomaterials 2016, 92,
71−80.
(17) Uddin, M. J.; Crews, B. C.; Ghebreselasie, K.; Daniel, C. K.;
Kingsley, P. J.; Xu, S.; Marnett, L. J. Targeted imaging of cancer by
fluorocoxib C, a near-infrared cyclooxygenase-2 probe. J. Biomed. Opt.
2015, 20, 50502.
(18) Uddin, M. J.; Crews, B. C.; Huda, I.; Ghebreselasie, K.; Daniel,
C. K.; Marnett, L. J. Trifluoromethyl fluorocoxib a detects
cyclooxygenase-2 expression in inflammatory tissues and human
tumor xenografts. ACS Med. Chem. Lett. 2014, 5, 446−450.
(19) Uddin, M. J.; Crews, B. C.; Ghebreselasie, K.; Marnett, L. J.
Design, synthesis, and structure-activity relationship studies of
fluorescent inhibitors of cycloxygenase-2 as targeted optical imaging
agents. Bioconjugate Chem. 2013, 24, 712−723.
(20) Uddin, M. J.; Crews, B. C.; Xu, S.; Ghebreselasie, K.; Daniel, C.
K.; Kingsley, P. J.; Banerjee, S.; Marnett, L. J. Antitumor Activity of
Cytotoxic Cyclooxygenase-2 Inhibitors. ACS Chem. Biol. 2016, 11,
3052−3060.
(21) Uddin, M. J.; Smithson, D. C.; Brown, K. M.; Crews, B. C.;
Connelly, M.; Zhu, F.; Marnett, L. J.; Guy, R. K. Podophyllotoxin
analogues active versus Trypanosoma brucei. Bioorg. Med. Chem. Lett.
2010, 20, 1787−1791.
(22) Uddin, M. J.; Crews, B. C.; Ghebreselasie, K.; Huda, I.;
Kingsley, P. J.; Ansari, M. S.; Tantawy, M. N.; Reese, J.; Marnett, L. J.
Fluorinated COX-2 inhibitors as agents in PET imaging of
inflammation and cancer. Cancer Prev. Res. 2011, 4, 1536−1545.
(23) Uddin, M. J.; Crews, B. C.; Ghebreselasie, K.; Tantawy, M. N.;
Marnett, L. J. [I]-Celecoxib Analogues as SPECT Tracers of
Cyclooxygenase-2 in Inflammation. ACS Med. Chem. Lett. 2011, 2,
160−164.
(24) Xu, S.; Uddin, M. J.; Banerjee, S.; Duggan, K.; Musee, J.; Kiefer,
J. R.; Ghebreselasie, K.; Rouzer, C. A.; Marnett, L. J. Fluorescent
indomethacin-dansyl conjugates utilize the membrane-binding
domain of cyclooxygenase-2 to block the opening to the active site.
J. Biol. Chem. 2019, 294, 8690−8698.
of the A. B. Hancock, Jr. Memorial Laboratory for Cancer
Research for critical reading and editing of the manuscript.
ABBREVIATIONS
■
CMP, N-[(5-carboxy-X-rhodaminyl)but-4-yl]-3-(1-(4-methox-
yphenyl)-5-(p-tolyl)-1H-pyrazol-3-yl)propenamide; COX, cy-
clooxygenase; PG, prostaglandin; PET, position emission
tomography; [18F]FDG, [18F]fluorodeoxyglucose; DMSO,
dimethyl sulfoxide
REFERENCES
■
(1) Rouzer, C. A.; Marnett, L. J. Non-redundant functions of
cyclooxygenases: oxygenation of endocannabinoids. J. Biol. Chem.
2008, 283, 8065−8069.
(2) Smith, W. L.; Garavito, R. M.; DeWitt, D. L. Prostaglandin
endoperoxide H synthases (cyclooxygenases)-1 and −2. J. Biol. Chem.
1996, 271, 33157−33160.
(3) Harris, R. E. Cyclooxygenase-2 (cox-2) and the inflammogenesis
of cancer. Subcell. Biochem. 2007, 42, 93−126.
(4) Hashemi Goradel, N.; Najafi, M.; Salehi, E.; Farhood, B.;
Mortezaee, K. Cyclooxygenase-2 in cancer: A review. J. Cell. Physiol.
2019, 234, 5683−5699.
(5) Wilson, A. J.; Fadare, O.; Beeghly-Fadiel, A.; Son, D. S.; Liu, Q.;
Zhao, S.; Saskowski, J.; Uddin, M. J.; Daniel, C.; Crews, B.; Lehmann,
B. D.; Pietenpol, J. A.; Crispens, M. A.; Marnett, L. J.; Khabele, D.
Aberrant over-expression of COX-1 intersects multiple pro-tumori-
genic pathways in high-grade serous ovarian cancer. Oncotarget 2015,
6, 21353−21368.
(6) Li, S.; Miner, K.; Fannin, R.; Carl Barrett, J.; Davis, B. J.
Cyclooxygenase-1 and 2 in normal and malignant human ovarian
epithelium. Gynecol. Oncol. 2004, 92, 622−627.
(7) Daikoku, T.; Wang, D.; Tranguch, S.; Morrow, J. D.; Orsulic, S.;
DuBois, R. N.; Dey, S. K. Cyclooxygenase-1 is a potential target for
prevention and treatment of ovarian epithelial cancer. Cancer Res.
2005, 65, 3735−3744.
(8) Gupta, R. A.; Tejada, L. V.; Tong, B. J.; Das, S. K.; Morrow, J.
D.; Dey, S. K.; DuBois, R. N. Cyclooxygenase-1 is overexpressed and
promotes angiogenic growth factor production in ovarian cancer.
Cancer Res. 2003, 63, 906−911.
(9) Motz, G. T.; Santoro, S. P.; Wang, L. P.; Garrabrant, T.; Lastra,
R. R.; Hagemann, I. S.; Lal, P.; Feldman, M. D.; Benencia, F.; Coukos,
G. Tumor endothelium FasL establishes a selective immune barrier
promoting tolerance in tumors. Nat. Med. 2014, 20, 607−615.
(10) Trabert, B.; Ness, R. B.; Lo-Ciganic, W. H.; Murphy, M. A.;
Goode, E. L.; Poole, E. M.; Brinton, L. A.; Webb, P. M.; Nagle, C. M.;
Jordan, S. J.; Risch, H. A.; Rossing, M. A.; Doherty, J. A.; Goodman,
M. T.; Lurie, G.; Kjaer, S. K.; Hogdall, E.; Jensen, A.; Cramer, D. W.;
Terry, K. L.; Vitonis, A.; Bandera, E. V.; Olson, S.; King, M. G.;
Chandran, U.; Anton-Culver, H.; Ziogas, A.; Menon, U.; Gayther, S.
A.; Ramus, S. J.; Gentry-Maharaj, A.; Wu, A. H.; Pearce, C. L.; Pike,
M. C.; Berchuck, A.; Schildkraut, J. M.; Wentzensen, N. Aspirin,
nonaspirin nonsteroidal anti-inflammatory drug, and acetaminophen
use and risk of invasive epithelial ovarian cancer: a pooled analysis in
the Ovarian Cancer Association Consortium. J. Natl. Cancer Inst.
2014, 106, djt431.
(25) Cingolani, G.; Panella, A.; Perrone, M. G.; Vitale, P.; Di Mauro,
G.; Fortuna, C. G.; Armen, R. S.; Ferorelli, S.; Smith, W. L.; Scilimati,
A. Structural basis for selective inhibition of Cyclooxygenase-1 (COX-
1) by diarylisoxazoles mofezolac and 3-(5-chlorofuran-2-yl)-5-methyl-
4-phenylisoxazole (P6). Eur. J. Med. Chem. 2017, 138, 661−668.
(11) Rothwell, P. M.; Wilson, M.; Price, J. F.; Belch, J. F.; Meade, T.
W.; Mehta, Z. Effect of daily aspirin on risk of cancer metastasis: a
study of incident cancers during randomised controlled trials. Lancet
2012, 379, 1591−1601.
(12) Rusu, D.; Carlier, T.; Colombie, M.; Goulon, D.; Fleury, V.;
Rousseau, N.; Berton-Rigaud, D.; Jaffre, I.; Kraeber-Bodere, F.;
Campion, L.; Rousseau, C. Clinical and Survival Impact of FDG PET
in Patients with Suspicion of Recurrent Ovarian Cancer: A 6-Year
Follow-Up. Front. Med. 2015, 2, 46.
(13) Lee, S. I.; Catalano, O. A.; Dehdashti, F. Evaluation of
gynecologic cancer with MR imaging, 18F-FDG PET/CT, and PET/
MR imaging. J. Nucl. Med. 2015, 56, 436−43.
F
DOI: 10.1021/acsmedchemlett.9b00280
ACS Med. Chem. Lett. XXXX, XXX, XXX−XXX