ARTICLE IN PRESS

Applied Radiation and Isotopes 67 (2009) 2019–2024

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Applied Radiation and Isotopes

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Synthesis of carbon-11 labeled celecoxib derivatives as new candidate PET

radioligands for imaging of inflammation
Mingzhang Gao a, Min Wang a, Kathy D. Miller b, Gary D. Hutchins a, Qi-Huang Zheng a,
a
Department of Radiology, Indiana University School of Medicine, 1345 West 16th Street, L3-208, Indianapolis, IN 46202, USA
b
Department of Medicine, Indiana University School of Medicine, Indianapolis, IN 46202, USA

a r t i c l e in fo abstract

Article history: Cyclooxygenase (prostaglandin endoperoxide synthase or COX) enzyme represents a particularly
Received 6 April 2009 attractive target in inflammation processes for the development of both therapeutic agents and imaging
Accepted 15 July 2009 agents. This study was designed to develop new radioligands for imaging of inflammation using the
biomedical imaging technique positron emission tomography (PET). Carbon-11 labeled celecoxib
Keywords: derivatives, [11C]methyl 2-(4-(5-p-tolyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenylsulfonamidooxy)
Positron emission tomography acetate ([11C]6e), [11C]methyl 2-methyl-2-(4-(5-p-tolyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenylsul-
Cyclooxygenase fonamidooxy)propanoate ([11C]6f), [11C]methyl 2-(4-(5-(4-methoxyphenyl)-3-(trifluoromethyl)-1H-
Celecoxib derivatives pyrazol-1-yl)phenylsulfonamidooxy)acetate ([11C]6g), and [11C]methyl 2-methyl-2-(4-(5-(4-methoxy-
Radioligands
phenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenylsulfonamidooxy)propanoate ([11C]6h), were prepared
Inflammation
by O-[11C]methylation of their corresponding precursors using [11C]CH3OTf under basic condition and
Imaging
isolated by a simplified solid-phase extraction (SPE) method in 50–60% radiochemical yields based on
[11C]CO2 and decay corrected to end of bombardment (EOB). The overall synthesis time from EOB was
15–20 min, the radiochemical purity was 499%, and the specific activity at end of synthesis (EOS) was
111–185 GBq/mmol.
& 2009 Elsevier Ltd. All rights reserved.

1. Introduction
The enzyme cyclooxygenase (prostaglandin endoperoxide
synthase or COX) catalyses the biosynthesis of prostaglandin,
prostacyclin and thromboxane from arachidonic acid (Anzini
et al., 2008). Three COX isozymes including COX-1, COX-2 and
COX-3 are actually described (Prabhakaran et al., 2005). COX-1 is
present in healthy tissues and responsible for the thrombogensis
and the homeostasis. COX-2 is almost undetectable under physio-
logic conditions and mainly induced by inflammatory stimuli. COX-
3 is located in the central nervous system and could be the
pharmacological target of acetaminophen. COX-2 over expression
is associated with inflammation processes and cancer progression
and contributes to the pathogenesis of several types of cancers
such as breast cancer and prostate cancer and neurodegenerative
diseases such as Alzheimer’s disease and Parkinson’s disease
(Kuge et al., 2006; Prabhakaran et al., 2005). COX-2 provides an
attractive target for the development of therapeutic agents. It was
proposed that a selective inhibitor of COX-2 would be a better
approach to treat inflammatory conditions, since the traditional
non-steroidal anti-inflammatory drugs (NSAIDs) are non-selective
 Corresponding author. Tel.: +1 317 278 4671; fax: +1 317 278 9711.
E-mail address: qzheng@iupui.edu (Q.-H. Zheng).
COX inhibitors that inhibit all COX enzymes. Two major classes of
compounds have been developed as COX-2 selective inhibitors:
the first one is the methanesulfonamide compounds like NS-398
and the second is ‘‘coxib’’ family such as celecoxib (Julemont et al.,
2004) (Fig. 1). However, it is important to note that some COX-2
selective inhibitors have been associated with significant
gastrointestinal side effects (Szabo et al., 2008). For example,
rofecoxib (Vioxx) has been withdrawn from the market due to the
enhanced risk for cardiovascular diseases. To improve the therapy
with fewer side effects, recently a novel series of celecoxib
derivatives have been designed and synthesized as more potent
anti-inflammatory agents (Szabo et al., 2008). COX-2 also provides
a particularly attractive target for the development of imaging
agents, and several papers have reported the synthesis and
preliminary evaluation of radiolabeled COX-2 inhibitors for
imaging of COX-2 using the biomedical imaging technique
positron emission tomography (PET) (de Vries et al., 2003;
McCarthy et al., 2002; Prabhakaran et al., 2005, 2007; Wust
et al., 2005). However, the potential of these radioligands for
imaging COX-2 expression remains unclear. In addition, most of
these COX-2 radioligands are labeled with positron emitting
radionuclide fluorine-18 (de Vries et al., 2003; McCarthy et al.,
2002; Prabhakaran et al., 2007; Wust et al., 2005). Recent efforts
have turned to develop selective COX-2 radioligands labeled with
another positron emitting radionuclide carbon-11, because some

0969-8043/$ - see front matter & 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.apradiso.2009.07.022
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2020 M. Gao et al. / Applied Radiation and Isotopes 67 (2009) 2019–2024

of fluorine-18 radioligands resulted in de[18F]fluorination in vivo
(Prabhakaran et al., 2005, 2007). We are interested in the
development of enzyme-based and/or receptor-based PET
imaging agents. In our previous works, we have described the
synthesis of carbon-11 labeled sulfonanilide analogs of NS-398 as
potential PET cancer imaging agents (Wang et al., 2007). In the
present study, we report the design, synthesis and labeling of a
series of celecoxib derivatives, [11C]methyl 2-(4-(5-p-tolyl-3-
(trifluoromethyl)-1H-pyrazol-1-yl)phenylsulfonamidooxy)acetate
([11C]6e), [11C]methyl 2-methyl-2-(4-(5-p-tolyl-3-(trifluoromethyl)-
1H-pyrazol-1-yl)phenylsulfonamidooxy)propanoate ([11C]6f), [11C]
methyl 2-(4-(5-(4-methoxyphenyl)-3-(trifluoromethyl)-1H-pyra-
zol-1-yl)phenylsulfonamidooxy)acetate ([11C]6g), and [11C]methyl
2-methyl-2-(4-(5-(4-methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazol-
1-yl)phenylsulfonamidooxy)propanoate ([11C]6h), as new candi-
date PET radioligands for imaging of inflammation.
2. Results and discussion
2.1. Chemistry
The chemistry is straightforward. Celecoxib was used as a
model compound for the design of a novel series of celecoxib
derivatives (Ahlstrom et al., 2007; Szabo et al., 2008). Celecoxib
O the transformation. The yield can be improved by adding some
H 2N
R3 S water in the reaction. The intermediate 4 was transferred to
O sulfonyl chloride 5 using phosphorus pentachloride in
O 2N O N
N CF3 R2
NH
O S O derivative 6 in 40–60% yield. The literature procedure (Szabo
CH3
R1 H 3C
Celecoxib
NS-398
Fig. 1. COX-2 inhibitors NS-398 and celecoxib.
contains a fairly rigid core structure with three apparent
interaction points: R1, R2, and R3 (Fig. 1). We designed new
celecoxib derivatives with slight chemical modifications in the R1
and R3 positions and without any change in the R2 position. These
newly designed target compounds were expected to be potent
anti-inflammatory agents with comparable anti-inflammatory
activity to their parent compounds and improved metabolic
properties, favorable lipophilicity to penetrate the blood–brain
barrier (BBB) and O-methyl position amenable to labeling with
carbon-11 (Ahlstrom et al., 2007; Gao et al., 2008; Liu et al., 2005;
Szabo et al., 2008). Moreover, we and other groups have reported a
methyl ester prodrug of matrix metalloproteinase (MMP) inhi-
bitor has better uptake in tumor than its acid parent drug
(Furumoto et al., 2003; Zheng et al., 2003). Synthesis of celecoxib
derivative acid precursors and methyl ester standards was
accomplished using a modification of the previously reported
procedures (Ahlstrom et al., 2007; Szabo et al., 2008). As depicted
in Scheme 1, Claisen condensation of the acetophenone 1 with
trifluoroacetic acid ethyl ester under strong base provided
diketone 2 in 92–93% yield. Two different bases NaH (Szabo
et al., 2008) and NaOMe (Ahlstrom et al., 2007) were tested in
the reaction, and NaH was proved to be a better base for the
reaction, which will give 2 at reliable high yield. Compound 2 was
reacted with p-hydrazino-benzene-sulfonic acid 3 through a
regioselective transformation to form 1,5-diarylpyrazole 4 in
81–86% yield. The solubility of compound 3 affected the yield of
dichloromethane with catalytic amount of DMF in 86–90% yield.
Compound 5 was reacted with aminooxy-carboxylic acid
or methyl ester in dichloromethane or DMF with Et3N and
4-dimethylaminopyridine (DMAP) to give its corresponding
et al., 2008) for the preparation of 6 gave poor yield (o30%) in our
hands. Therefore, we modified the literature procedure (two-
phase reaction under dioxane/water) to a single phase reaction by
using DMF as a solvent, Et3N as a base and DMAP as a catalyst.

O
HO
NHNH2 S
O
CF3CO2Et N
O NaH/THF, 0 oC O N CF3
HO3S 3
X X O
EtOH, H2O/HCl
CH3
1a, X=Me reflux
2a, X=Me, 93% CF3
1b, X=OMe 2b, X=OMe, 92%
X 4a, X=Me, 86%
4b, X=OMe, 81%

PCl5, CH2Cl2
O O reflux
RONH Cl
S S
O O
N N
N CF3 6a, X=Me, R=CH 2CO2H CF3
RONH2 HCl, DMF N
6b, X=Me, R=C(CH 3)2CO2H Et3N, DMAP, rt
6c, X=OMe, R=CH 2CO2H
6d, X=OMe, R=C(CH 3)2CO2H
6e, X=Me, R=CH 2CO2CH3
X 6f, X=Me, R=C(CH 3)2CO2CH3 X 5a, X=Me, 90%
6g, X=OMe, R=CH 2CO2CH3
5b, X=OMe, 86%
6h, X=OMe, R=C(CH 3)2CO2CH3
40-60%

Scheme 1. Synthesis of celecoxib derivative precursors and standards.

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M. Gao et al. / Applied Radiation and Isotopes 67 (2009) 2019–2024 2021

O O
RONH RONH
S S
O O
N
11
CH3OTf, CH3CN N
CF3 N CF3
N
2 N NaOH, 80 oC

X X
6a, X=Me, R=CH 2CO2H [11C]6e, X=Me, R=CH 2CO211CH3
6b, X=Me, R=C(CH 3)2CO2H [11C]6f, X=Me, R=C(CH 3)2CO211CH3
6c, X=OMe, R=CH 2CO2H [11C]6g, X=OMe, R=CH 2CO211CH3
6d, X=OMe, R=C(CH 3)2CO2H [11C]6h, X=OMe, R=C(CH 3)2CO211CH3
50-60%

Scheme 2. Synthesis of carbon-11 labeled celecoxib derivatives.

This modification significantly improved the yield. Compounds
6a–d are acid precursors for carbon-11 labeling, and compound
6e–h are methyl ester reference standards.
2.2. Radiochemistry
497%. The radiochemical purity of the target tracers was 499%
determined by radio-HPLC through g-ray (PIN diode) flow
detector, and the chemical purity of the target tracers was
495% determined by reversed-phase HPLC through UV flow
detector.

Synthesis of carbon-11 labeled celecoxib derivatives [11C]6e–h

is indicated in Scheme 2. Acid precursors 6a–d were labeled by a
reactive [11C]methylating agent, [11C]methyl triflate ([11C]CH3OTf)
(Jewett, 1992; Mock et al., 1999) prepared from [11C]CO2,
under basic conditions (2 N NaOH) in acetonitrile through the
O-[11C]methylation and isolated by a simplified solid-phase
extraction (SPE) method (Yoder et al., 2009; Zheng and
Mulholland, 1996) to provide target tracers [11C]6e–h in 50–60%
radiochemical yields, decay corrected to end of bombardment
(EOB), based on [11C]CO2. The large polarity difference between
the acid precursor and the labeled O-methylated ester product
permitted the use of SPE technique for purification of the labeled
product from the radiolabeling reaction mixture. Either a C-18
Plus Sep-Pak cartridge (disposable) or a semi-prep C-18 guard
cartridge column (repeat use) was used in SPE purification
technique. The crude reaction mixture was treated with sodium
bicarbonate and loaded onto the cartridge by gas pressure. Any
non-reacted acid precursor was actually converted to the
corresponding sodium salt, and any non-reacted [11C]CH3OTf
was actually hydrolyzed to [11C]CH3OH, which would not stick to
the C-18 Sep-Pak. The cartridge was washed with water to remove
non-reacted [11C]CH3OTf, acid precursor and reaction solvent, and
then the final labeled product was eluted with ethanol. Overall
synthesis time was 15–20 min from EOB. SPE technique is fast,
efficient and convenient and works very well for the O-methylated
ester tracer purification using the acid precursor for radiolabel-
ing (Yoder et al., 2009; Zheng and Mulholland, 1996). The
radiosynthesis was performed in an automated multi-purpose
11
C-radiosynthesis module, allowing measurement of specific
activity during synthesis (Mock et al., 2005a, b). The specific
activity was estimated in a range of 111–185 GBq/mmol at the end
of synthesis (EOS) based on other compounds produced using the
same targetry conditions which have been measured by the on-
the-fly technique (Mock et al., 2005a; Zheng et al., 2007). The
actual measurement of specific activity at EOS by analytical HPLC
method (Zheng and Mock, 2005) is in agreement with this
estimation. Chemical purity and radiochemical purity were
determined by analytical HPLC method (Zheng and Mock, 2005).
The chemical purity of the precursors and reference standards was
3. Materials and methods
3.1. General
All commercial reagents and solvents from Aldrich and Sigma
were used without further purification. [11C]CH3OTf was prepared
according to a literature procedure (Mock et al., 1999). Melting
points were determined on a MEL-TEMP II capillary tube
apparatus and were uncorrected. 1H NMR spectra were recorded
on Varian Gemini 2000 200 MHz FT-NMR and Bruker Avance II
500 MHz NMR spectrometers using tetramethylsilane (TMS) as an
internal standard. Chemical shift data for the proton resonances
were reported in parts per million (ppm, d scale) relative to
internal standard TMS (d 0.0), and coupling constants (J) were
reported in hertz (Hz). Low resolution mass spectra (LRMS) were
obtained using a Bruker Biflex III MALDI-Tof mass spectrometer,
and high resolution mass spectra (HRMS) were obtained using a
Thermo MAT 95XP-Trap spectrometer. Chromatographic solvent
proportions are indicated as volume: volume ratio. Thin-layer
chromatography (TLC) was run using Analtech silica gel GF
uniplates (5  10 cm2). Plates were visualized under UV light.
Preparative TLC was run using Analtech silica gel UV 254 plates
(20  20 cm2). Normal phase flash column chromatography was
carried out on EM Science silica gel 60 (230–400 mesh) with a
forced flow of the indicated solvent system in the proportions
described below. All moisture- and/or air-sensitive reactions were
performed under a positive pressure of nitrogen maintained by a
direct line from a nitrogen source.
Analytical HPLC was performed using a Prodigy (Phenomenex)
5 mm C-18 column, 4.6  250 mm; 3:1:1 CH3CN:MeOH:20 mM, pH
6.7 phosphate (buffer solution) mobile phase; flow rate 1.5 mL/
min; and UV (254 nm) and g-ray (PIN diode) flow detectors. C-18
Plus Sep-Pak cartridges were obtained from Waters Corporate
Headquarters, Milford, MA. Semi-prep C-18 guard cartridge
column 1 1 cm was obtained from E.S. Industries, Berlin, NJ,
and part number 300121-C18-BD 10 mm. Sterile Millex-GS
0.22 mm vented filter unit was obtained from Millipore Corpora-
tion, Bedford, MA.
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3.2. 4,4,4-Trifluoro-1-(4-methylphenyl)butane-1,3-dione (2a)
4-Methyl-acetophenone 1a (20.0 g, 149 mmol) was dissolved in
dry THF (300 mL) under nitrogen atmosphere and NaH (7.15 g,
298 mmol) was added in portions maintaining the temperature
between 5 and 0 1C. After stirring at this temperature for 30 min,
ethyl trifluoroacetate (31.75 g, 224 mmol) was added and the
reaction mixture was allowed to stir at room temperature (rt) for
6 h. The reaction mixture was evaporated, added with ice-water,
acidified with HCl (1 N) to pH 6, and extracted with ethyl acetate
(3  100 mL). The combined organic layer was washed with water
(50 mL), dried over MgSO4, and concentrated under reduced
pressure. The solid residue was washed with hexane and dried
under high vacuum to provide a solid mass, which was re-
dissolved in dichloromethane and dried under high vacuum to
give 2a (31.87 g, 93%) as a light brown solid, Rf ¼ 0.25 (25% EtOAc/
hexanes), mp 40–42 1C. 1H NMR (CDCl3): d 2.44 (s, 3H, CH3), 6.54
(s, 1H, CH ¼ ), 7.28 (d, J ¼ 8.0 Hz, 2H, Ph-H), 7.83 (d, J ¼ 8.0 Hz, 2H,
Ph-H). MS (EI): 229 ([M–H]+, 91%), 159 (100%).
3.3. 4,4,4-Trifluoro-1-(4-methoxyphenyl)butane-1,3-dione (2b)
Compound 2b was prepared from 4-methoxy-acetophenone
1b using the procedure described for 2a as a light brown solid in
92% yield, Rf ¼ 0.24 (25% EtOAc/hexanes), mp 48–49 1C. 1H NMR
(CDCl3): d 3.90 (s, 3H, OCH3), 6.50 (s, 1H, CH ¼ ), 6.96
(d, J ¼ 8.8 Hz, 2H, Ph-H), 7.91 (d, J ¼ 9.0 Hz, 2H, Ph-H). MS (ESI):
247 ([M+H]+, 100%).
3.4. 4-(5-p-Tolyl-3-(trifluoromethyl)-1H-pyrazol-1-
yl)benzenesulfonic acid (4a)
Compound 2a (6.00 g, 26 mmol) was added into a solution of p-
hydrazino-benzenesulfonic acid 3 (5.64 g, 30 mmol) in ethanol
(320 mL) and HCl (6 N, 8.6 mL) and water (40 mL). The mixture
was heated to reflux and stirred for 20 h. Then the reaction
mixture was evaporated under reduced pressure, extracted with
ethyl acetate (4  120 mL), washed with brine, dried over MgSO4,
filtered, and evaporated to dryness. The residue was purified by
column chromatography on silica gel (12% MeOH/CH2Cl2) to
afford 4a (8.54 g, 86%) as a white solid, Rf ¼ 0.14 (10% MeOH/
CH2Cl2), mp4280 1C. 1H NMR (DMSO-d6): d 2.30 (s, 3H, CH3), 7.14
(s, 1H, CH ¼ ), 7.18–7.19 (m, 4H, Ph-H), 7.26 (d, J ¼ 8.8 Hz, 2H, Ph-
H), 7.62 (d, J ¼ 8.8 Hz, 2H, Ph-H). MS (ESI): 383 ([M+H]+, 100%).
3.5. 4-(5-(4-Methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-
yl)benzenesulfonic acid (4b)
Compound 4b was prepared from 2b using the procedure
described for 4a as a white solid in 81% yield, Rf ¼ 0.12 (10%
MeOH/CH2Cl2), mp 298–300 1C. 1H NMR (DMSO-d6): d 3.76 (s, 3H,
CH3), 6.94 (d, J ¼ 8.8 Hz, 2H, Ph-H), 7.10 (s, 1H, CH ¼ ), 7.20
(d, J ¼ 9.0 Hz, 2H, Ph-H), 7.24 (d, J ¼ 8.6 Hz, 2H, Ph-H), 7.63
(d, J ¼ 8.4 Hz, 2H, Ph-H). MS (ESI): 399 ([M+H]+, 100%).
3.6. 4-(5-p-Tolyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzene-1-
sulfonyl chloride (5a)
Phosphorus pentachloride (2.2 g, 10.5 mmol) was added part-
wise to a suspension of compound 4a (2.52 g, 6.6 mmol) in
dichloromethane (60 mL). Then DMF (1 mL) was added to the
reaction solution. The resulting solution was refluxed for 8 h,
and the mixture was concentrated under vacuum. Ice-water
was added to the residue and extracted with ethyl acetate
(2  120 mL), dried over MgSO4, and evaporated to dryness. The
residue was crystallized from hexane and ethyl acetate mixture to
give 5a (2.38 g, 90%) as a white solid, mp 85–87 1C. 1H NMR
(CDCl3): d 2.40 (s, 3H, CH3), 6.75 (s, 1H, CH ¼ ), 7.10–7.25 (m, 4H,
Ph-H), 7.56 (d, J ¼ 8.8 Hz, 2H, Ph-H), 7.99 (d, J ¼ 8.8 Hz, 2H, Ph-H).
3.7. 4-(5-(4-Methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-
yl)benzene-1-sulfonyl chloride (5b)
Compound 5b was prepared from 4b using the procedure
described for 5a as a white solid in 86% yield, mp 109–110 1C. 1H
NMR (CDCl3): d 3.85 (s, 3H, CH3), 6.73 (s, 1H, CH ¼ ), 6.90
(d, J ¼ 8.8 Hz, 2H, Ph-H), 7.15 (d, J ¼ 8.6 Hz, 2H, Ph-H), 7.56
(d, J ¼ 8.8 Hz, 2H, Ph-H), 7.99 (d, J ¼ 8.8 Hz, 2H, Ph-H).
3.8. General procedure for preparation of celecoxib derivative 6
The benzenesulfonyl chloride compound 5 (0.5 mmol) was
added to a solution of aminooxy-carboxylic acid hydrochloride or
aminooxy-carboxylic acid methyl ester (0.6 mmol) and Et3N
(101 mg, 1.0 mmol) in dichloromethane (40 mL). DMAP was
added to this solution (61 mg, 0.5 mmol) and stirred at rt for 5 h.
Then the reaction mixture was concentrated under vacuum. The
residue was added with water and extracted with ethyl acetate
(2  80 mL), dried over MgSO4, evaporated, and purified by
column chromatography on silica gel (5% MeOH/CH2Cl2) or (10%
EtOAc/hexanes) to afford 6 as a white solid in 40–60% yield.
2-(4-(5-p-Tolyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenylsu-
lfonamidooxy)acetic acid (6a), Rf ¼ 0.73 (20% MeOH/CH2Cl2), mp
221–223 1C. 1H NMR (CDCl3): d 2.31 (s, 3H, CH3), 4.37 (s, 2H, CH2),
6.96 (s, 1H, CH ¼ ), 7.20–7.24 (m, 4H, Ph-H), 7.56–7.57 (m, 2H,
Ph-H), 7.95–7.96 (m, 2H, Ph-H), 9.9 (s, 1H). MS (ESI): 456 ([M+H]+,
100%).
2-Methyl-2-(4-(5-p-tolyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)
phenylsulfonamidooxy)propanoic acid (6b), Rf ¼ 0.80 (20% MeOH/
CH2Cl2), mp 93–95 1C. 1H NMR (CDCl3): d 1.33 (s, 6H, 2xCH3),
2.35 (s, 3H, CH3), 6.72 (s, 1H, CH ¼ ), 7.06 (d, J ¼ 7.5 Hz, 2H, Ph-H),
7.14 (d, J ¼ 7.5 Hz, 2H, Ph-H), 7.43 (d, J ¼ 7.5 Hz, 2H, Ph-H), 7.85
(d, J ¼ 8.0 Hz, 2H, Ph-H), 8.01 (s, 1H). MS (ESI): 484 ([M+H]+,
100%).
2-(4-(5-(4-Methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-
yl)phenylsulfonamidooxy)acetic acid (6c), Rf ¼ 0.68 (20% MeOH/
CH2Cl2), mp 108–110 1C. 1H NMR (CDCl3): d 3.75 (s, 3H, CH3), 4.30
(s, 2H, CH2), 4.39 (s, 1H), 6.87 (s, 1H, CH ¼ ), 6.91 (d, J ¼ 7.2 Hz, 2H,
Ph-H), 7.20 (d, J ¼ 8.2 Hz, 2H, Ph-H), 7.53 (d, J ¼ 8.2 Hz, 2H, Ph-H),
7.89 (d, J ¼ 7.2 Hz, 2H, Ph-H). MS (ESI): 472 ([M+H]+, 100%). HRMS
(CI): calcd for C19H17O6N3F3S1 ([M+H]+), 472.0785; found
472.0767.
2-(4-(5-(4-Methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-
yl)phenylsulfonamidooxy)-2-methylpropanoic acid (6d), Rf ¼ 0.77
(20% MeOH/CH2Cl2), mp 97–99 1C. 1H NMR (DMSO-d6): d 1.47
(s, 6H, 2  CH3), 3.83 (s, 3H, CH3), 6.78 (s, 1H, CH ¼ ), 6.90
(d, J ¼ 8.0 Hz, 2H, Ph-H), 7.19 (d, J ¼ 8.5 Hz, 2H, Ph-H), 7.51
(d, J ¼ 8.0 Hz, 2H, Ph-H), 7.65 (s, 1H), 7.94 (d, J ¼ 8.0 Hz, 2H, Ph-H),
8.82 (s, 1H). MS (ESI): 499 (M+, 100%). HRMS (ESI): calcd for
C21H21O6N3F3S1 ([M+H]+), 500.1103; found 500.1081.
Methyl 2-(4-(5-p-tolyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)-
phenylsulfonamidooxy)acetate (6e), Rf ¼ 0.67 (34% EtOAc/hex-
anes), mp 137–139 1C. 1H NMR (CDCl3): d 2.39 (s, 3H, CH3), 3.81
(s, 3H, CH3), 4.51 (s, 2H, CH2), 6.74 (s, 1H, CH ¼ ), 7.11 (d, J ¼ 8.0 Hz,
2H, Ph-H), 7.18 (d, J ¼ 8.0 Hz, 2H, Ph-H), 7.48 (d, J ¼ 9.0 Hz, 2H,
Ph-H), 7.79 (s, 1H), 7.89 (d, J ¼ 9.0 Hz, 2H, Ph-H). MS (ESI): 470
([M+H]+, 100%). HRMS (ESI): calcd for C20H19O5N3F3S1 ([M+H]+),
470.0998; found 470.1003.
Methyl 2-methyl-2-(4-(5-p-tolyl-3-(trifluoromethyl)-1H-pyra-
zol-1-yl)phenylsulfonamidooxy)propanoate (6f), Rf ¼ 0.71 (34%
 ARTICLE IN PRESS
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EtOAc/hexanes), mp 53–55 1C. 1H NMR (CDCl3): d 1.46 (s, 6H,
2  CH3), 2.38 (s, 3H, CH3), 3.72 (s, 3H, CH3), 6.74 (s, 1H, CH ¼ ),
7.09 (d, J ¼ 8.0 Hz, 2H, Ph-H), 7.16 (d, J ¼ 8.4 Hz, 2H, Ph-H), 7.33
(s, 1H), 7.48 (d, J ¼ 8.8 Hz, 2H, Ph-H), 7.89 (d, J ¼ 8.8 Hz, 2H, Ph-H).
MS (ESI): 498 ([M+H]+, 100%). HRMS (ESI): calcd for C22H22O5N3-
F3S1Na ([M+Na]+), 520.1130; found 520.1104.
Methyl 2-(4-(5-(4-methoxyphenyl)-3-(trifluoromethyl)-1H-
pyrazol-1-yl)phenylsulfonamidooxy)acetate (6g), Rf ¼ 0.53 (34%
EtOAc/hexanes), mp 106–108 1C. 1H NMR (CDCl3): d 3.76 (s, 3H,
CH3), 3.83 (s, 3H, CH3), 4.51 (s, 2H, CH2), 6.71 (s, 1H, CH ¼ ), 6.87
(d, J ¼ 8.8 Hz, 2H, Ph-H), 7.13 (d, J ¼ 8.4 Hz, 2H, Ph-H), 7.49
(d, J ¼ 8.8 Hz, 2H, Ph-H), 7.80 (s, 1H), 7.89 (d, J ¼ 8.6 Hz, 2H, Ph-H).
MS (ESI): 486 ([M+H]+, 100%). HRMS (ESI): calcd for C20H19O6N3
F3S1 ([M+H]+), 486.0947; found 486.0957.
Methyl 2-(4-(5-(4-methoxyphenyl)-3-(trifluoromethyl)-1H-
pyrazol-1-yl)phenylsulfonamidooxy)-2-methylpropanoate (6h),
Rf ¼ 0.60 (34% EtOAc/hexanes), mp 80–82 1C. 1H NMR (CDCl3): d
1.46 (s, 6H, 2  CH3), 3.72 (s, 3H, CH3), 3.83 (s, 3H, CH3), 6.72
(s, 1H, CH ¼ ), 6.87 (d, J ¼ 8.8 Hz, 2H, Ph-H), 7.13 (d, J ¼ 8.8 Hz,
2H, Ph-H), 7.33 (s, 1H), 7.49 (d, J ¼ 8.8 Hz, 2H, Ph-H), 7.88
(d, J ¼ 8.6 Hz, 2H, Ph-H). MS (ESI): 514 ([M+H]+, 100%). HRMS
(ESI): calcd for C22H23O6N3F3S1 ([M+H]+), 514.1260; found
514.1254.
3.9. General procedure for preparation of target tracers, [11C]methyl
2-(4-(5-p-tolyl-3-(trifluoromethyl)-1H-pyrazol-1-
yl)phenylsulfonamidooxy)acetate ð½11 C6eÞ, [11C]methyl 2-methyl-2-
(4-(5-p-tolyl-3-(trifluoromethyl)-1H-pyrazol-1-
yl)phenylsulfonamidooxy)propanoate ð½11 C6fÞ, [11C]methyl 2-(4-(5-
(4-methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-
yl)phenylsulfonamidooxy)acetate ð½11 C6gÞ, and [11C]methyl 2-
methyl-2-(4-(5-(4-methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazol-
1-yl)phenylsulfonamidooxy)propanoate ð½11 C6hÞ
11 14 11
[ C]CO2 was produced by the N(p,a) C nuclear reaction in
small volume (9.5 cm3) aluminum gas target (CTI) from 11 MeV
proton cyclotron on research purity nitrogen (+1% O2) in a
Siemens radionuclide delivery system (Eclipse RDS-111). The acid
precursor 6a, 6b, 6c, or 6d (0.1–0.3 mg) was dissolved in CH3CN
(300 mL). To this solution was added 2 N NaOH (2 mL). The mixture
was transferred to a small reaction vial. No-carrier-added (high
specific activity) [11C]CH3OTf that was produced by the gas-phase
production method (Mock et al., 1999) from [11C]CO2 through
[11C]CH4 and [11C]CH3Br with silver triflate (AgOTf) column was
passed into the reaction vial at rt until radioactivity reached a
maximum (2 min), and then the reaction vial was isolated and
heated at 80 1C for 3 min. The contents of the reaction vial were
diluted with NaHCO3 (0.1 M, 1 mL). The reaction tube was
connected to either a C-18 Plus Sep-Pak cartridge or a semi-prep
C-18 guard cartridge column. The labeled product mixture
solution was passed onto the cartridge for SPE purification by
gas pressure. The cartridge was washed with H2O (2  3 mL), and
the aqueous washing was discarded. The product was eluted from
the column with EtOH (2  3 mL), and then passed onto a rotatory
evaporator. The solvent was removed by evaporation under
vacuum. The labeled product [11C]6e, [11C]6f, [11C]6g, or [11C]6h
was formulated with saline, sterile-filtered through a sterile
vented Millex-GS 0.22 mm cellulose acetate membrane and
collected into a sterile vial. Total radioactivity was assayed and
the total volume was noted for tracer dose dispensing. The overall
synthesis time including SPE purification and formulation was
15–20 min. The radiochemical yields decay corrected to EOB, from
[11C]CO2, were 50–60%. Retention times in the analytical HPLC
system were: tR 6a ¼ 2.83 min, tR 6e ¼ 4.68 min, tR [11C]6e ¼ 4.68
min; tR 6b ¼ 2.75 min, tR 6f ¼ 4.95 min, tR [11C]6f ¼ 4.95 min; tR
2023
6c ¼ 3.14 min, tR 6g ¼ 5.56 min, tR [11C]6g ¼ 5.56 min; and tR
6d ¼ 3.25 min, tR 6h ¼ 5.47 min, tR [11C]6h ¼ 5.47 min.
4. Conclusions
An efficient and convenient synthesis of new radioligands,
carbon-11 labeled celecoxib derivatives, has been well developed.
The synthetic methodology employed classical organic chemistry
such as Claisen condensation, regioselective transformation,
sulfonylation and amidating reaction to prepare a series of new
celecoxib derivative precursors and standard compounds. The
target radioligands were prepared by O-[11C]methylation of their
corresponding acid precursors using a reactive [11C]methylating
agent, [11C]CH3OTf, and isolated by a simplified SPE purification
procedure in high radiochemical yields, short overall synthesis
time, and great specific radioactivities. These chemistry results
combined with the reported in vitro and in vivo biological data
(Ahlstrom et al., 2007; Prabhakaran et al., 2005, 2007; Szabo et al.,
2008) encourage further in vivo biological evaluation of new
carbon-11 labeled celecoxib derivatives, as candidate radioligands
to image inflammation.
Acknowledgments
This work was partially supported by the Susan G. Komen for
the Cure, Breast Cancer Research Foundation and Indiana
Genomics Initiative (INGEN) of Indiana University, which is
supported in part by Lilly Endowment Inc. The authors would
like to thank Dr. Bruce H. Mock and Barbara E. Glick-Wilson for
their assistance in production of [11C]CH3OTf.
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