Chemical preparation, biological evaluation and 3D-QSAR of ethoxysulfuron derivatives as novel antifungal agents targeting acetohydroxyacid synthase

Article abstract of DOI:10.1016/j.ejmech.2018.11.005

Accetohydroxyacid synthase (AHAS) is the first enzyme involved in the biosynthetic pathway of branched-chain amino acids. Earlier gene mutation of Candida albicans in a mouse model suggested that this enzyme is a promising target of antifungals. Recent studies have demonstrated that some commercial AHAS-inhibiting sulfonylurea herbicides exerted desirable antifungal activity. In this study, we have designed and synthesized 68 novel ethoxysulfulron (ES) derivatives and evaluated their inhibition constants (K_i) against C. albicans AHAS and cell based minimum inhibitory concentration (MIC) values. The target compounds 5-1, 5-10, 5-22, 5-31 and 5-37 displayed stronger AHAS inhibitions than ES did. Compound 5-1 had the best K_i of 6.7 nM against fungal AHAS and MIC values of 2.5 mg/L against Candida albicans and Candica parapsilosis after 72 h. A suitable nematode model was established here and the antifungal activity of 5-1 was further evaluated in vivo. A possible binding mode was simulated via molecular docking and a comparative field analysis (CoMFA) model was constructed to understand the structure-activity relationship. The current study has indicated that some ES derivatives should be considered as promising hits to develop antifungal drugs with novel biological target.

Full text of DOI:10.1016/j.ejmech.2018.11.005

Accepted Manuscript
Chemical preparation, biological evaluation and 3D-QSAR of ethoxysulfuron
derivatives as novel antifungal agents targeting acetohydroxyacid synthase
Ren-Jun Wu, Tongtong Ren, Jie-Yu Gao, Li Wang, Qilin Yu, Zheng Yao, Guo-Qing
Song, Wei-Bin Ruan, Cong-Wei Niu, Fu-Hang Song, Li-Xin Zhang, Mingchun Li, Jian-
Guo Wang
PII:
S0223-5234(18)30959-0
DOI:
https://doi.org/10.1016/j.ejmech.2018.11.005
EJMECH 10862
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 8 October 2018
Revised Date: 1 November 2018
Accepted Date: 3 November 2018
Please cite this article as: R.-J. Wu, T. Ren, J.-Y. Gao, L. Wang, Q. Yu, Z. Yao, G.-Q. Song,
W.-B. Ruan, C.-W. Niu, F.-H. Song, L.-X. Zhang, M. Li, J.-G. Wang, Chemical preparation,
biological evaluation and 3D-QSAR of ethoxysulfuron derivatives as novel antifungal agents
targeting acetohydroxyacid synthase, European Journal of Medicinal Chemistry (2018), doi: https://
doi.org/10.1016/j.ejmech.2018.11.005.
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ACCEPTED MANUSCRIPT
Chemical preparation, biological evaluation and 3D-QSAR of ethoxysulfuron derivatives as novel antifungal agents
targeting acetohydroxyacid synthase
a 1
b 1
c,d 1
a
b
a
a
b
a
Ren-Jun Wu , Tongtong Ren , Jie-Yu Gao , Li Wang , Qilin Yu , Zheng Yao , Guo-Qing Song , Wei-Bin Ruan , Cong-Wei Niu , Fu-Hang
c
c,e**
b,***
a,*
Song , Li-Xin Zhang , Mingchun Li
, Jian-Guo Wang

ACCEPTED MANUSCRIPT
Chemical preparation, biological evaluation and 3D-QSAR of
ethoxysulfuron derivatives as novel antifungal agents targeting
acetohydroxyacid synthase
a 1
b 1
c,d 1
a
b
a
Ren-Jun Wu , Tongtong Ren , Jie-Yu Gao , Li Wang , Qilin Yu , Zheng Yao ,
a
b
a
c
c,e**
Guo-Qing Song , Wei-Bin Ruan , Cong-Wei Niu , Fu-Hang Song , Li-Xin Zhang
,
b,***
a,*
Mingchun Li
, Jian-Guo Wang
a. State-Key Laboratory and Research Institute of Elemento-Organic Chemistry,
College of Chemistry, Nankai University, Tianjin 300071, China
b. Key Laboratory of Molecular Microbiology and Technology, Ministry of
Education, College of Life Sciences, Nankai University, Tianjin, 300071, China.
c. State Key Laboratory of Microbial Resources and CAS Key Laboratory of
Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese
Academy of Sciences, Beijing, 100101, China.
d. School of Food and Biological Engineering, Hefei University of Technology, Hefei,
Anhui 230009, China
e. State Key Laboratory of Bioreactor Engineering, East China University of Science
and Technology, Shanghai 200237, China.
1
These three authors contributed equally to this work.
*
corresponding author. E-mail address: nkwjg@nankai.edu.cn (J.-G. Wang)
**
corresponding author. E-mail address: zhanglixin@im.ac.cn (L.-X. Zhang)
***
corresponding author. E-mail address: nklimingchun@163.com(M. Li)
Abstract: Accetohydroxyacid synthase (AHAS) is the first enzyme involved in the
biosynthetic pathway of branched-chain amino acids. Earlier gene mutation of
Candida albicans in a mouse model suggested that this enzyme is a promising target
of antifungals. Recent studies have demonstrated that some commercial
AHAS-inhibiting sulfonylurea herbicides exerted desirable antifungal activity. In this
study, we have designed and synthesized 68 novel ethoxysulfulron (ES) derivatives
and evaluated their inhibition constants (K ) against C. albicans AHAS and cell based
i
minimum inhibitory concentration (MIC) values. The target compounds 5-1, 5-10,
5
-22, 5-31 and 5-37 displayed stronger AHAS inhibitions than ES did. Compound 5-1
had the best K of 6.7 nM against fungal AHAS and MIC values of 2.5 mg/L against
i
Candida albicans and Candica parapsilosis after 72h. A suitable nematode model was

ACCEPTED MANUSCRIPT
established here and the antifungal activity of 5-1 was further evaluated in vivo. A
possible binding mode was simulated via molecular docking and a comparative field
analysis (CoMFA) model was constructed to understand the structure-activity
relationship. The current study has indicated that some ES derivatives should be
considered as promising hits to develop antifungal drugs with novel biological target.
Keywords: acetohydroxyacid synthase, ethoxysulfuron derivative, enzyme inhibition,
antifungal agents, nematode model.
1
. Introduction
Fungal infections such as candidiasis, aspergillosis and cryptococcosis have become a
serious threat towards the health of AIDS patients, cancer sufferers and organ
transplant recipients, which has caused increasing rates for both morbidity and
mortality [1-2]. It is estimated that invasive fungal infections affect approximately 1.2
billion individuals globally, associated with nearly 1.5 million annual deaths [3-4].
Presently, systemic therapy against such diseases mainly depends on a few classes of
antifungal agents, including azoles, echinocandins, polyenes, allylamines and
pyrimidine analogs [5-6], among which fluconazole (FCZ) and amphotericin B (AMB)
are the most representative examples. With the continuous use of these medicines,
resistance has emerged and evolved as a significant problem [7]. Currently, the
discovery of antifungal compounds mainly relies on a small number of targets such as
ergosterol synthesis, ergosterol disruptors, glucan synthesis, and nucleic acid
synthesis [8]. No new drugs have been introduced to the market for the treatment of
invasive fungal infections for years since the approval of anidulafungin. It is hence an
important mission to explore new target enzyme to develop different antifungal drugs.
Acetohydroxyacid synthase (AHAS, EC 2.2.1.6) is the first enzyme involved in the
biosynthetic pathway of the branched-chain amino acids (BCAAs), which is essential
in plants and microbes but absent in mammals [9]. Indeed, AHAS has been a
successful target in the research of some commercial herbicides for several decades
[
10-19]. Recently, some compounds have been identified for antituberculosis activity
targeting AHAS, suggesting that AHAS is a potential target for the research of
antimicrobial chemotherapy [20-24]. Earlier in a murine infection model, when the
gene for AHAS was mutated in Cryptococcus neoformans and Candida albicans,

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these fungi became avirulent or significantly attenuated in virulence due to the lack of
BCAAs, showing that AHAS is a promising target for antifungal agents [25, 26]. A
study in 2013 demonstrated that some sulfonylurea herbicides such as chlorimuron
ethyl (CE) and ethoxysulfuron (ES) are potent inhibitors of C. albicans AHAS [27].
Another report in the same year conducted by Novartis also showed that some AHAS
inhibitors have antifungal activity in cell based assay [28]. More lately, it has been
shown that CE had antifungal activity in a mouse model for the first time [29]. From
another point of view, a few sulfonylurea compounds were also reported to possess
desirable activity against some plant fungi as potential agricultural fungicides [30].
The sulfonylurea herbicides were developed in the 1970s, with ultralow dosage,
environmentally benign and non-toxicity advantages [31, 32]. For instance, the oral
LD for CE is >4000 mg/kg in rats, while its dermal LD is >2000 mg/kg, meaning
5
0
50
that this compound is fairly safe towards mammals [33]. Some special sulfonylurea
herbicide such as monosulfuron ester (MSE) was also invented by some academic
institution [10]. Although dozens of sulfonylurea compounds are excellent herbicides,
only a few of them are good antifungal inhibitors [27-29]. In fact, the sulfonylureas
had been famous as antidiabetic drugs before their agrochemical use as herbicides.
Early in the 1950s, carbutamide was developed as the first generation of sulfonylurea
antibiabetic drugs [34]. In the 1990s, glimepiride came to market as the third
generation for the medication of type II diabetes [35]. These examples imply that
herbicidal sulfonylureas might also have the potential of medicinal use. The chemical
structures of FCZ, AMB, anidulafungin, CE, ES, MSE, carbutamide and glimepiride
are illustrated in Fig. 1.
Herein, in an effort to discover novel antifungal compounds, we have designed and
synthesized a series of ES derivatives, the antifungal potencies of which were
validated against both fungal AHAS and fungal pathogens in cell based assay. 5 of the
total 68 target compounds displayed better inhibition constants against C. albicans
AHAS than that of ES, among which compound 5-1 has the best K value of 6.7 nM
i
against the enzyme and MIC values of 2.5~5 mg/L against different fungal strains.
Moreover, 5-1 showed desirable in vivo antifungal activity in a nematode model,
comparable to that of CE, ES and FCZ. Finally, the binding mode of 5-1 with fungal
AHAS was generated from molecular docking and a predictive comparative field

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analysis (CoMFA) model was also constructed to quantitatively understand the
structure-activity relationships. The present study will furnish valuable guidance for
the discovery of antifungal drugs to cure fungal infectious diseases.
2
. Results and Discussion
2
.1 Chemistry of the target compounds
As shown in Scheme 1, various substituted phenols 1 reacted with sulfuryl chloride
isocyanate in toluene using a refluxing process to give oil intermediate 2, which was
further reacted with different 4,6-substituted 2-aminopyrimidines 3 in acetonitrile at
room temperature. In most cases, product 4 was hard to precipitate out for easy
preparation, therefore saturated sodium bicarbonate was added to the mixture and the
ES derivatives were readily obtained in their corresponding sodium salts 5 as a simple
precipitate in desirable yields. Compound 4 can be prepared backwards by
acidification of compound 5 in water in a simple manner. For comparison, we also
synthesized the sodium salt of ES. Generally, other families of sulfonylureas
precipitate easily in acetonitrile without having to make them as salts. Nevertheless,
for these ES derivatives, they do not precipitate smoothly in their acidified products.
As it can be seen later, that the sodium salts of the ES derivatives exhibited equal
AHAS inhibitions and antifungal activities with the corresponding acidified
compounds 4, the target compounds in this study were therefore prepared in their
sodium salts 5 instead.
The chemical structures of all the target compounds (5-1 to 5-68) were fully
1
13
characterized and confirmed by H NMR, C NMR and HRMS (The original
spectrogram are provided in the supplementary data). The melting points of the title
compounds were also determined. Their molecular structures are listed in Table 1.
In order to further confirm their chemical structures, 5-57 and its according acidified
product 4-57 were successfully recrystallized in ethyl ether/acetone to give colorless
crystals suitable for single-crystal X-ray diffraction after we had attempted several
cases. These two crystal structures are shown in Fig. 2. The salt form of 5-57 is a
dimer in the crystal and the sodium ions can be seen clearly; In contrast, for the
acidified product 4-57 there is only one sulfonylurea molecule in the crystal. The
crystal structure of 4-57 is somewhat interesting in that the pyrimidine rings possess

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alternative poses as a duplex crystal.
2
.2 Inhibition of C. albicans AHAS
All the synthesized sulfonylureas were subjected to C. albicans AHAS inhibition
assay and the determined K vulaes are listed in Table 1. Although these compounds
i
are basically similar ES derivatives, their inhibition constants cover a large range
from 6.7 nM to >50000 nM. For comparison, sodium salt of ES and 4-1 (acidified
product of 5-1) were also measured their fungal AHAS inhibitory potency. As can be
seen, 4-1 has equal potency to 5-1 while ES has similar K values with its sodium salt,
i
which reveals that the ES derivative salts generally have same inhibitory potency
compared with their corresponding acidified products. Compounds 5-1, 5-10, 5-22,
5
-31 and 5-37 have better K values than that of ES (21.6 nM), among which 5-1 has
i
the best K of 6.7±0.9 nM. On the contrary, Compound 5-8, 5-64, 5-66, 5-67 and 5-68
i
are so weak that their K values could not be determined at 50000 nM. It should be
i
noted that another control MSE is a very weak inhibitor of C. albicans AHAS, with a
determined K value of 16820 nM, which is 778 times and 2510 times weaker than
i
those of ES and 5-1. This indicated again that not all good plant AHAS inhibitors are
good inhibitors of fungal AHAS. The inhibition curve of 5-1 is presented in Fig. 3.
2
.3 Possible mode of action and structure-activity relationships
The crystal structure of C. albicans AHAS in complex with CE is an ideal starting
point to predict the binding mode of 5-1 (also 4-1) [29]. From the molecular docking
result (Fig. 4a), compound 5-1 (green) superposes with CE (cyan) fairly well,
suggesting that the computational simulation is considerably reasonable. Because the
binding pocket of AHAS is in fact in the interface formed by two different subunits,
symbol A and B were used here to distinguish residues from different subunits.
Basically, the pyrimidine ring in the molecule forms a π-π stacking interaction with
W582(B), and several intermolecular hydrogen bonds were found between the
inhibitor and neighboring residues, namely K247(A) and R376(B). Besides W582(B),
M578(B), R376(B), D375(B), A191(A), P188(A) and K247(A), which have already
been described clearly [24], the surrounding residues also include G112(A), L115(A),
V187(A), A196(A), F197(A), Q198(A), H351(B), V579(B) and A653(B), via multiple
hydrophobic interactions with the inhibitor, as depicted from the Fig. 4a. Compound
5
-1 inhabits in a cavity formed by the nearby residues, as shown in Fig. 4b.

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With respect to the structure-activity relationships, for the AHAS inhibitory activity,
from a simple two-dimensional (2D) view, an ortho substituent in R position
1
generally provides desirable AHAS inhibition, yet if this substituent is at meta or para
position, the inhibition becomes much weaker. For example, 5-1 and 5-8 have same
formula and the only difference between them is the R position, K of 5-8 could not
1
i
be measured at 50000 nM while K of 5-1 is at nanomolar level. When R is an ester
i
1
group in the ortho position, a –COOC H group offers the best activity, as shown by
2
5
5
-1, 5-10, 5-22, 5-31, 5-37 and 5-57. For the variations of R and R , two –OCH
2 3 3
substituents provide the best inhibition, a halogen atom substitution of –OCH gives
3
slightly weaker activity, while two –CH groups at same position decrease the activity
3
greatly.
Comparative field analysis (CoMFA) is a tool to generate 3D contour models to
quantitatively analyze the structure-activity relationships (QSAR) of bioactive
compounds by steric and electrostatic contributions [36-39]. The conformation of 5-1
from molecular docking was used to construct and chemical structures of molecular
database. Besides 5-8, 5-64, 5-66, 5-67 and 5-68, whose K values were unavailable
i
here, compounds 5-38, 5-43 and 5-58 were also excluded because they were statistical
2
outliers in the training set. The remaining training set gave a leave-one-out q of 0.540
when the number of optimum components was 8. For the non-crossvalidated result as
2
this condition, r was 0.908 with a standard error of estimate of 0.315 and F values of
6
2.777 (The experimental activity versus the predicted activity of the compounds
were listed in the supplemental data). The steric contribution was 54.9% and the
electrostatic contribution was 45.1%. Compound 5-1 was used to depict the steric and
electrostatic contour maps, as shown in Fig. 5. For the steric contour map (Fig. 5a), a
bulky group is favorable for better AHAS inhibition in the green contour region and
such a group is likely to decrease the activity in the yellow contour space. This is in a
general agreement with the 2D structure-activity relationship explanation, that the two
–
OCH groups in the heterocycle offer better enzyme inhibition and a steric group at
3
the ortho position (R ) is favorable, however if the ortho group is too large, then the
1
activity will be weak. For the electrostatic contour map (Fig. 5b), in the blue contour
region, an increase in the positive charge will provide an increase of activity, whereas
in the red contour region, negative charge is likely to enhance the AHAS inhibitory

ACCEPTED MANUSCRIPT
activity. Either the ester group or the ester group in the ortho position fits the map in
view of electrostatic field.
2
.3 Cell based antifungal activity
The biological data are listed in Table 2, which were measured at 24 h against C.
albicans standard isolate sc5314. For assay using RPMI 1640 media, only compounds
5
-1, 5-4, 5-31, 5-34, 5-37 and 5-40 display moderate to weak activity. When switched
to YNB media, compounds 5-1, 5-4, 5-5, 5-6, 5-10, 5-31, 5-34, 5-35, 5-36, 5-37, 5-40,
-41, 5-42, 5-43 and 5-65 exhibit different levels of inhibition. Obviously, the
antifungal activity in YNB media enhanced significantly. The MICs for ES, 5-1 and
-4 change from 12.5 mg/L to 0.625 mg/L , from 25 mg/L to 1.25 mg/L and from 50
5
5
mg/L to 2.5 mg/L, respectively, 20-fold improvement for each of them. Similar result
had also been reported for CE, with 2.3 times change for MICs against C. albicans at
conditions with and without BCAAs for different fungi, and for other Candida strains,
there is a >20 times shift for CE [29]. Meanwhile, for FCZ and AMB, MICs remain
the same at both conditions, 0.25 mg/L for the former and 1.25 mg/L for the later. The
1
5 compounds with preliminary antifungal activity are general potent inhibitors of
fungal AHAS. Among these compounds, 5-4, 5-6, 5-35, 5-36, 5-42, 5-43 and 5-65
possess K values of 100~700 nM, while 5-1, 5-5, 5-10, 5-31, 5-34, 5-37, 5-40 and
i
5
-41 have K values of <100 nM. It should be pointed out that 5-1 is the best inhibitor
i
of C. albicans AHAS and also the most potent one for the cell based assay. On the
other hand, ES is not as active as 5-1 for AHAS inhibition, but its antifungal activity
is twofold of the potency for 5-1. Similarly, 5-4 is a much weaker inhibitor of C.
albicans AHAS compared with 5-1, yet its antifungal activity is just two times weaker
at this condition. This result means that besides AHAS inhibition, factors of
absorption, distribution, metabolism, and excretion (ADME) should also be taken into
account for further design of such compounds.
Based on these results, 5-1 and 5-4 was further investigated its antifungal
susceptibility using different fungal strains. 4-1, sodium salt of ES, FCZ and AMB
were also tested for comparison. It can be seen from Table 3 that 5-1 and 4-1 have
nearly the same activity, so do ES and its sodium salt, suggesting that the sodium salts
here are suitable for investigation of their biological activity in this study. 17# and g5
are two clinically isolated FCZ-resistant strains of C. albicans, and the sulfonylurea

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inhibitors also exhibit good activity towards these fungi. After 24h, FCZ lost it
activity, as reported previously [40]. 5-1 and ES became twofold weaker or remain the
same upon the five fungal strains when observed at 72h, the MICs of which are
1
.25~5.0 mg/L. However, the activity of 5-4 decreased dramatically at 72. As another
control, the activity of AMB does not change from 24h to 72h, with MIC of 1.25
mg/L against all the C. albicans strains and 2.5 mg/L against S. cerevisiae SC
XH1549 and C. parapsilosis ATCC22019 (Table 3).
2
.4 In vivo antifungal activity in nematode model
Although the ES derivatives exhibited promising antifungal activity in the cell based
assay, it is hard to tell the in vivo efficiency of these compounds. The recent study by
Guddat et al reported a mouse model to evaluate the antifungal activity of CE,
however this experiment is time-consuming and expensive [29]. Therefore a simple
but reliable nematode model is needed to determine the preliminary in vivo activity of
the sulfonylureas before a murine infection assay is carried out. In this study a
modified nematode model [41-43] was established to evaluate the in vivo antifungal
ability of the target compounds based on an elementary animal model. Since these
compounds take effect by inhibiting the biosynthesis of branched-chain amino acids,
YNB medium was used to avoid the influence of inner branched-china amino acids
that were available under other medium conditions. Without being infected, the
nematodes in other medium conditions can survive for at least six days; however, in
the absence of necessary amino acids, the nematodes die of starvation at 48h under
YNB condition. Obviously, the survival period for infected nematodes at this
condition is shorter than 48h. After several times of experiments, we found that there
was a significant difference between the infected nematodes with and without being
treated by inhibitors at 24h for YNB condition. Thereupon in this study the mortality
rate was measured at 24h when 20nM of 5-1, ES, CE and FCZ had been added into
the medium. Fig 6a and 6b show the in vivo efficiency of the tested compounds. For
the control, there was a mortality rate of 17% for the infected nematodes. Excitingly,
for 5-1, ES, CE and FCZ, the mortality rates were 7%, 3%, 8% and 5%, respectively,
all of which displayed drop compared with the control. The present data suggested
that commercial sulfonylurea herbicides ES and CE had therapeutic effects against C.
albicans at the nematode model, for which ES displayed better efficacy than CE. As a
newly synthesized compound, 5-1 was a bit weaker than ES but slightly better than

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CE, indicating its potential for further research. As a clinically used agent, FCZ
exhibited similar potency at this model. The nematode model here has provided a
cheap and quick technique for the determination of the in vivo antifungal activity of
the sulfonylurea compounds, which is simple but more meaningful than the cell based
assay.
Resistance is likely to occur and become a serious problem associated with the
overuse of drugs or herbicides. Strong resistance has been observed in multiple weed
species when commercial sulfonylurea herbicides have been overused [44]. In most
cases the resistance is due to residue mutation in the binding site, such as P197, A205,
W574, and S653 (A. thaliana AHAS numbering). Due to the similar binding modes of
sulfonylureas with either plant AHAS or fungal AHAS [29], the same problem is an
essential factor to keep in mind for the future design of antifungal sulfonylureas.
Basically, care should be taken to minimize interaction with these residues or
alternatively to provide structural flexibility for the sulfonylurea to adjust its
conformation or location so that potent inhibition toward a mutated form of the
enzyme can be maintained [27]. Some successful examples have been reported by
Yang et al [16-18], using conformationally flexible inhibitors to overcome this
problem. It is suggested that a similar protocol might be used for the design of new
antifungal ES derivatives.
3
. Conclusion
Due to the limited choices and the resistance problem of clinical antifungal agents, the
discovery and invention of novel inhibitors that act on a different target enzyme has
become an important and urgent task. However, it is a challenge and difficulty to
achieve this goal. In this paper, dozens of ES derivatives were newly designed and
synthesized, which were fully evaluated by fungal AHAS inhibition and cell based
assay. It was found that the sodium salts of ES derivatives have equal potent activities.
Compound 5-1 was discovered as a possible hit for further research, which has similar
activity with ES. 5-1 and ES also showed desirable activity at the nematode model,
indicating that they could be considered as candidates for further research. Molecular
simulations of CoMFA model and molecular docking have given valuable information
to understand the structure-activity relationships and probable binding action. Based
on the current results, fungal AHAS bound with 5-1 will be studied by

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co-crystallization and its antifungal capability will also be evaluated using a mouse
model soon, together with that of ES. The present research will provide valuable ideas
for the development of new family of antifungal medicines with a novel mode of
action.
4
. Experimental section
4
.1. General synthesis and instruments
The chemistry route for the target compounds is depicted in Scheme 1. Most of the
intermediates 1 and 3 were purchased from Alfa-Aesar, Sigma-Aldrich, TCI,
Apichemical and Chemieliva, which were all >95% purity grade. Two of the
intermediates 1 (R =OCH CH I for 1-a and R =COOCH CH I for 1-b) were not
1
2
2
1
2
2
commercially available and the synthesis for them are detailed below. Other related
starting materials were procured from J&K Chemical, Accela ChemBio, Xinzun
Chemical and some local chemical suppliers from Tianjin. All solvents and liquid
reagents were dried in advance using standard methods and distilled before use.
Melting points were determined using an RT-4 melting apparatus and were
1
13
uncorrected. H NMR spectra and C NMR were obtained using a Bruker Avance
00 MHz spectrometer. The chemical shift values (d) for the NMR spectra were
4
reported as parts per million (ppm), using deuterated chloroform (CDCl ) or dimethyl
3
sulfoxide (DMSO-d ) as the solvent and tetramethylsilane (TMS) as an internal
6
reference standard. Mass spectra were recorded on an Ionspect FT-MS 7.0T LC/mass
detector instrument. Single-crystal X-ray diffraction analyses were performed on a
Bruker Smart 1000 CCD diffractometer.
4
.2. Synthesis of intermediate 1-a and 1-b
1
5 mmol 1-a' (or 1-b') and 20 mmol sodium iodide were added to 50 mL acetone, the
reaction mixture was stirred for 24 h under reflux. After that the reaction was cooled
down to room temperature and the resulting sodium chloride was filtered out. The
filtrate was extracted by dichloromethane/water and organic phase was collected.
Then dichloromethane was removed and 1-a (or 1-b) was obtained.
4
.2.1. 2-iodoethyl 2-hydroxybenzoate (1-a)
1
Yield 70%; yellow oil; H NMR (400 MHz, DMSO-d ) δ 10.39 (s,1H,OH), 7.83 (d, J
6
=
=
7.8 Hz, 1H, ArH), 7.55 (t, J = 6.9 Hz, 1H, ArH), 7.03 – 6.96 (m, 2H, ArH), 4.57 (t, J
6.2 Hz, 2H, COOCH ), 3.55 (t, J = 6.2 Hz, 2H, CH I).
2
2

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4
.2.2. 2-(2-iodoethoxy)phenol
1
Yield 65%; m.p. 80-82 °C; yellow solid; H NMR (400 MHz, DMSO-d ) δ 9.03 (s,
6
1
H, OH), 6.93 (d, J = 7.7 Hz, 1H, ArH), 6.81 (d, J = 3.9 Hz, 2H, ArH), 6.73 (d, J =
.6 Hz, 1H, ArH), 4.22 (t, J = 6.4 Hz, 2H, COOCH ) , 3.48 (t, J = 6.3 Hz, 2H, CH I).
7
2
2
4
.3. Synthesis of the target ES derivatives
For the synthesis of 2, the solution of sulfurisocyanatidic chloride (7.2 mmol) in 20
mL toluene was added to the solution of 1 (6.0 mmol) in 20 mL toluene dropwise at
room temperature. The reactant was heated to 140 °C and then the reaction proceeded
for 18h under reflux. Subsequently, the mixture was cooled down to room
temperature and remaining sulfurisocyanatidic chloride was removed under reduced
pressure, together with the solvent. Without further purification, the resulting yellow
oil 2 was dissolved in 10 mL anhydrous acetonitrile and after that it was added slowly
to 5 mmol of 3, which was also dissolved in 10 mL anhydrous acetonitrile beforehead
in ice bath. After stirring for 24 h at room temperature, acetonitrile was removed
under reduced pressure and saturated sodium bicarbonate was added to product 4.
Product 5 precipitated easily and it was further purified by recrystallization from
petroleum ether/acetone in 1:1 ratio in high yields. 15% hydrochloric acid was added
to aqueous solution of 5 under stirring and corresponding acidified product 4
precipitated out easily in high yields.
4
.3.1. Sodium ((4,6-dimethoxypyrimidin-2-yl)carbamoyl)((2-(ethoxycarbonyl)phenoxy)
sulfonyl)amide (5-1) and ethyl 2-((N-((4,6-dimethoxypyrimidin-2-yl)carbamoyl)
sulfamoyl)oxy)benzoate (4-1)
1
5
8
7
-1 Yield 89%; m.p.: 220-222 °C; white solid; H NMR (400 MHz, DMSO-d ), δ
6
.40 (s, 1H, CONH) , 7.60 (d, J = 7.6 Hz, 1H, ArH) ,7.53 – 7.44 (m, 2H, ArH), 7.25 –
.19 (m, 1H, ArH), 5.68 (s, 1H, Prim-H), 4.23 (q, J = 7.1 Hz, 2H, OCH ), 3.82 (s, 6H,
2
13
OCH ), 1.28 (t, J = 7.1 Hz, 3H, CH CH ); C NMR (101 MHz, DMSO-d ), δ 171.9,
3
2
3
6
1
66.3, 158.4, 155.8, 150.6, 132.8, 130.4, 126.5, 124.9, 123.9, 82.1, 61.1, 54.2, 14.4;
+
+
HRMS (ESI) m/z: calculated for C H N NaO S (M-Na +2H ) 427.0919, found
1
6
17
4
8
4
27.0922.
-1 Yield 96%; m.p.: 187-189 °C; white solid; H NMR (400 MHz, CDCl ), δ 13.00
1
4
3
(
s, 1H, SO NH), 7.96 (dd, J = 7.7, 1.7 Hz, 1H, ArH), 7.60 – 7.50 (m, 1H, ArH), 7.48 –
2
7
.35 (m, 2H, ArH), 5.77 (s, 1H, Prim-H), 4.37 (q, J = 7.1 Hz, 2H, OCH ), 3.84 (s, 6H,
2
OCH ), 1.39 (t, J = 7.1 Hz, 3H, CH CH ).
3
2
3

ACCEPTED MANUSCRIPT
4
.3.2. Sodium ((4,6-dimethoxypyrimidin-2-yl)carbamoyl)((2-(propoxycarbonyl)
phenoxy)sulfonyl)amide (5-2)
Yield 80%; m.p.: 215-217 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.41 (s,
1
6
1
H, CONH), 7.60 (d, J = 8.6 Hz, 1H, ArH), 7.49 (t, J = 7.5 Hz, 2H, ArH), 7.24 – 7.20
(
m, 1H, ArH), 5.68 (s, 1H, Prim-H), 4.13 (t, J = 6.7 Hz, 2H, OCH ), 3.82 (s, 6H,
2
13
OCH ), 1.68 (m, 2H, OCH CH ), 0.94 (t, J = 7.4 Hz, 3H, CH CH ); C NMR (101
3
2
2
2
3
MHz, DMSO-d ), δ 171.9, 166.4, 158.4, 155.6, 150.7, 132.8, 130.4, 126.4, 124.8,
6
1
23.9, 82.2, 66.6, 54.1, 21.9, 10.9; HRMS (ESI) m/z: calculated for C H N NaO S
17 19 4 8
+
+
(
M-Na +2H ) 441.1075, found 441.1073.
4
.3.3. Sodium ((4,6-dimethoxypyrimidin-2-yl)carbamoyl)((2-((2-iodoethoxy)carbonyl)
phenoxy)sulfonyl)amide (5-3)
Yield 94%; m.p.: 191-193 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.46 (s,
1
6
1
H, CONH), 7.66 (d, J = 9.1 Hz, 1H, ArH), 7.54 (t, J = 7.7 Hz, 1H, ArH), 7.46 (d, J =
.8 Hz, 1H, ArH), 7.26 (t, J = 7.2 Hz, 1H, ArH), 5.69 (s, 1H, Prim-H), 4.44 (t, J = 6.9
7
13
Hz, 2H, OCH ), 3.83 (s, 6H, OCH ), 3.47 (t, J = 6.9 Hz, 2H, CH CH I); C NMR
2
3
2
2
(
101 MHz, DMSO-d ), δ 171.9, 165.5, 158.4, 155.5, 150.9, 133.2, 130.5, 125.9, 124.9,
6
1
24.1, 82.3, 65. 4, 54.2, 2.8; HRMS (ESI) m/z: calculated for C H IN NaO S
16 16 4 8
+
+
(
M-Na +2H ) 552.9885, found 552.9879.
4
.34. Sodium ((4,6-dimethoxypyrimidin-2-yl)carbamoyl)((2-(2-iodoethoxy)phenoxy)
sulfonyl)amide (5-4)
Yield 84%; m.p.:134-136 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.54 (s,
1
6
1
H, CONH), 7.44 (d, J = 8.2 Hz, 1H, ArH), 7.04 (m, 2H, ArH), 6.95 – 6.88 (m, 1H,
ArH), 5.68 (s, 1H, Prim-H), 4.24 (t, J = 6.9 Hz, 2H, OCH ), 3.82 (s, 6H, OCH ), 3.46
2
3
1
3
(
t, J = 6.5 Hz, 2H, CH CH I); C NMR (101 MHz, DMSO-d ), δ 171.9, 158.4, 155.8,
2 2 6
1
50.4, 142.3, 125.7, 123.7, 121.9, 116.6, 82.2, 70.7, 54.2, 4.0; HRMS (ESI) m/z:
+
+
calculated for C H IN NaO S (M-Na +2H ) 524.9936, found 524.9931.
1
5
16
4
7
4
.3.5. Sodium ((4,6-dimethoxypyrimidin-2-yl)carbamoyl)((2-((2,2,2-trifluoroethoxy)
carbonyl)phenoxy)sulfonyl)amide (5-5)
Yield 86%; m.p.: 173-174 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.43 (s,
1
6
1
H, CONH), 7.66 – 7.55 (m, 2H, ArH), 7.49 (d, J = 7.0 Hz, 1H, ArH), 7.28 (t, J = 7.4
Hz, 1H, ArH), 5.69 (s, 1H, Prim-H), 4.95 – 4.87 (m, 2H, CH CF ), 3.82 (s, 6H,
2
3
13
OCH ); C NMR (101 MHz, DMSO-d ), δ 172.0, 164.5, 158.4, 155.9, 150.9, 130.6,
3
6
1
25.2, 124.7, 124.4, 122.7, 82.5, 60.7, 54.2; HRMS (ESI) m/z: calculated for
+
+
C H F N NaO S (M-Na +2H ) 481.0636, found 481.0627.
16
14
3
4
8

ACCEPTED MANUSCRIPT
4
.3.6. Sodium ((4,6-dimethoxypyrimidin-2-yl)carbamoyl)((2-(isopropoxycarbonyl)
phenoxy)sulfonyl)amide (5-6)
Yield 85%; m.p.: 142-144 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.36 (s,
1
6
1
H, CONH), 7.57 (d, J = 9.0 Hz, 1H, ArH), 7.48 (q, J = 8.3 Hz, 2H, ArH), 7.24 – 7.18
(m, 1H, ArH), 5.68 (s, 1H, Prim-H), 5.06 (dt, J = 12.5, 6.2 Hz, 1H, OCH), 3.82 (s, 6H,
1
3
OCH ), 1.30 (d, J = 6.2 Hz, 6H, CH ); C NMR (101 MHz, DMSO-d ), δ 172.0,
3
3
6
1
65.8, 158.4, 156.1, 150.5, 132.7, 130.4, 126.8, 124.9, 123.8, 82.0, 68.6, 54.3, 22.0;
+
+
HRMS (ESI) m/z: calculated for C H N NaO S (M-Na +2H ) 441.1075, found
1
7
19
4
8
4
41.1080.
4
.3.7.
Sodium
((4,6-dimethoxypyrimidin-2-yl)carbamoyl)((2-(phenoxycarbonyl)
phenoxy)sulfonyl)amide (5-7)
Yield 78%; m.p.: 118-120 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.51 (s,
1
6
1
H, CONH), 7.79 (d, J = 7.7 Hz, 1H, ArH), 7.61 (t, J = 7.8 Hz, 1H, ArH), 7.49 (d, J =
8
.3 Hz, 1H, ArH), 7.47 – 7.24 (m, 6H, ArH), 5.70 (s, 1H, Prim-H), 3.82 (s, 6H, OCH3);
1
3
C NMR (101 MHz, DMSO-d ), δ 172.0, 165.1, 158.4, 156.3, 151.4, 150.8, 133.7,
6
1
31.0, 129.8, 126.3, 126.0, 125.4, 124.5, 122.6, 82.1, 54.3; HRMS (ESI) m/z:
+
+
calculated for C H N NaO S (M-Na +2H ) 475.0919, found 475.0922.
2
0
17
4
8
4
.3.8.
phenoxy)sulfonyl)amide (5-8)
Yield 71%; m.p.: 136-138 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.59 (s,
Sodium
((4,6-dimethoxypyrimidin-2-yl)carbamoyl)((4-(ethoxycarbonyl)
1
6
1
H, CONH), 7.92 (d, J = 8.7 Hz, 2H, ArH), 7.36 (d, J = 8.7 Hz, 2H, ArH), 5.69 (s, 1H,
Prim-H), 4.29 (q, J = 7.1 Hz, 2H, OCH ), 3.81 (s, 6H, OCH ), 1.30 (t, J = 7.1 Hz, 3H,
2
3
13
CH CH ); C NMR (101 MHz, DMSO-d ), δ 172.0, 165.7, 158.3, 156.6, 156.2, 130.9,
2
3
6
1
26.1, 121.6, 82.1, 61.1, 54.3, 14.6; HRMS (ESI) m/z: calculated for C H N NaO S
16 17 4 8
+
+
(
M-Na +2H ) 427.0919, found 427.0921.
4
.3.9. Sodium ((4-chloro-6-methoxypyrimidin-2-yl)carbamoyl)((2-(methoxycarbonyl)
phenoxy)sulfonyl)amide (5-9)
Yield 81%; m.p.: 226-228 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.02 (s,
1
6
1
H, CONH), 7.61 (d, J = 7.0 Hz, 1H, ArH), 7.55 – 7.44 (m, 2H, ArH), 7.23 (t, J = 7.4
13
Hz, 1H, ArH), 6.50 (s, 1H, Prim-H), 3.89 (s, 3H, COOCH ), 3.77 (s, 3H, OCH ); C
3
3
NMR (101 MHz, DMSO-d ), δ 171.1, 166.7, 160.2, 158.9, 155.6, 150.5, 133.0, 130.5,
6
1
26.2, 125.1, 124.0, 99.3, 54.7, 52.5; HRMS (ESI) m/z: calculated for
+
+
C H ClN NaO S (M-Na +2H ) 417.0267, found 417.0272.
14
13
4
7
4
.3.10. Sodium ((4-chloro-6-methoxypyrimidin-2-yl)carbamoyl)((2-(ethoxycarbonyl)

ACCEPTED MANUSCRIPT
phenoxy) sulfonyl)amide (5-10)
1
Yield 92%; m.p.: 212-214 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.99 (s,
6
1
H, CONH), 7.60 (d, J = 7.7 Hz, 1H, ArH), 7.49 (t, J = 15.8, 8.1 Hz, 2H, ArH), 7.23 (t,
J = 7.3 Hz, 1H, ArH), 6.50 (s, 1H, Prim-H), 4.23 (q, J = 7.1 Hz, 2H, OCH ), 3.89 (s,
2
13
3
H, OCH ), 1.28 (t, J = 7.1 Hz, 3H, CH ); C NMR (101 MHz, DMSO-d ), δ 171.0,
3 3 6
1
66.3, 160.1, 159.0, 155.3, 150.6, 132. 8, 130.4, 126.6, 124.9, 124.0, 99.2, 61.1, 54.6,
+
+
1
4.4; HRMS (ESI) m/z: calculated for C H ClN NaO S (M-Na +2H ) 431.0423,
15 14 4 7
found 431.0428.
4
.3.11. Sodium ((4-chloro-6-methoxypyrimidin-2-yl)carbamoyl)((2-(propoxycarbonyl)
phenoxy)sulfonyl) amide (5-11)
Yield 89%; m.p.: 199-201 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.98 (s,
1
6
1
H, CONH), 7.60 (d, J = 8.1 Hz, 1H, ArH), 7.49 (t, J = 7.0 Hz, 2H, ArH ), 7.22 (t, J =
7
.0 Hz, 1H, ArH), 6.49 (s, 1H, Prim-H), 4.13 (t, J = 6.7 Hz, 2H, OCH ), 3.88 (s, 3H,
2
13
OCH ), 1.68 (m, 2H, OCH CH ), 0.94 (t, J = 7.4 Hz, 3H, CH CH ); C NMR (101
3
2
2
2
3
MHz, DMSO-d ), δ 171.0, 166.4, 160.1, 158.9, 155.2, 150.7, 132.7, 130.4, 126.5,
6
1
24.8, 123.9, 99.2, 66.7, 54.6, 21.9, 10.9; HRMS (ESI) m/z: calculated for
+
+
C H ClN NaO S (M-Na +2H ) 445.0580, found 445.0582.
16
16
4
7
4
.3.12. Sodium ((4-chloro-6-methoxypyrimidin-2-yl)carbamoyl)((2-((2-iodoethoxy)
carbonyl)phenoxy) sulfonyl)amide (5-12)
1
Yield 79%; m.p.: 207-209°C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.03 (s,
6
1
H, CONH), 7.66 (d, J = 7.3 Hz, 1H, ArH), 7.58 – 7.51 (m, 1H, ArH), 7.46 (d, J = 8.1
Hz, 1H, ArH), 7.26 (t, J = 7.3 Hz, 1H, ArH), 6.50 (s, 1H, Prim-H), 4.44 (t, J = 6.9 Hz,
1
3
2
H, OCH ), 3.89 (s, 3H, OCH ), 3.47 (t, J = 6.8 Hz, 2H, CH I); C NMR (101 MHz,
2 3 2
DMSO-d ), δ 171.1, 165.5, 160.1, 158.9, 155.5, 150.6, 133.3, 130.6, 125.8, 125.1,
6
1
24.1, 99.4, 65.5, 54.7, 2.8; HRMS (ESI) m/z: calculated for C H ClIN NaO S
15 13 4 7
+
+
(
M-Na +2H ) 556.9390, found 556.9395.
4
.3.13. Sodium ((4-chloro-6-methoxypyrimidin-2-yl)carbamoyl)((2-(2-chloroethoxy)
phenoxy)sulfonyl)amide (5-13)
Yield 83%; m.p.: 123-125 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.10 (s,
1
6
1
9
3
1
H, CONH), 7.47 (d, J = 7.7 Hz, 1H, ArH), 7.05 (d, J = 5.4 Hz, 2H , ArH), 6.93 (t, J =
.4, 4.1 Hz, 1H, ArH), 6.49 (s, 1H, Prim-H), 4.24 (t, J = 5.6 Hz, 2H, OCH ), 3.95 –
2
1
3
.86 (m, 5H, CH Cl and OCH ); C NMR (101 MHz, DMSO-d ), δ 171.1, 160.1,
2
3
6
58.9, 155.5, 150.6, 142.3, 125.6, 123.6, 121.9, 116.5, 99.3, 70.2, 54.6, 43.3; HRMS
+
+
(
ESI) m/z: calculated for C H Cl N NaO S (M-Na +2H ) 437.0084, found
14 13 2 4 6

ACCEPTED MANUSCRIPT
4
37.0089.
.3.14. Sodium ((4-chloro-6-methoxypyrimidin-2-yl)carbamoyl)((2-(2-iodoethoxy)
4
phenoxy)sulfonyl)amide (5-14)
Yield 78%; m.p.: 113-115 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.11 (s,
1
6
1
1
3
1
H, CONH), 7.45 (d, J = 7.5 Hz, 1H, ArH), 7.09 – 7.03 (m, 2H, ArH), 6.95 – 6.90 (m,
H, ArH), 6.50 (s, 1H, Prim-H), 4.25 (t, J = 6.9 Hz, 2H, OCH ), 3.89 (s, 3H, OCH ),
2
3
1
3
.47 (t, J = 6.9 Hz, 2H, CH I); C NMR (101 MHz, DMSO-d ), δ 171.1, 160.2, 158.9,
2
6
55.7, 150.4, 142.2, 125.8, 123.9, 121.9, 116.6, 99.3, 70.7, 54.7, 3.9; HRMS (ESI)
+
+
m/z: calculated for C H ClIN NaO S (M-Na +2H ) 528.9441, found 528.9440.
1
4
13
4
6
4
.3.15.
Sodium
((4-chloro-6-methoxypyrimidin-2-yl)carbamoyl)((2-((2,2,2-
trifluoroethoxy)carbonyl)phenoxy)sulfonyl)amide (5-15)
1
Yield 83%; m.p.: 115-116°C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.04 (s,
6
1
H, CONH), 7.67 – 7.55 (m, 2H, ArH), 7.49 (d, J = 7.7 Hz, 1H, ArH), 7.28 (t, J = 7.1
Hz, 1H, ArH), 6.50 (s, 1H, Prim-H), 4.96 – 4.87 (m, 2H, CH CF ), 3.88 (s, 3H,
2
3
13
OCH ); C NMR (101 MHz, DMSO-d ), δ 171.1, 164.4, 160.2, 158.9, 155.7, 150.9,
3
6
1
33.8, 130.6, 128.7, 125.3, 124.7, 124.4, 99.3, 61.0, 54.6; HRMS (ESI) m/z:
+
+
calculated for C H ClF N NaO S (M-Na +2H ) 485.0141, found 485.0148.
1
5
11
3
4
7
4
.3.16. Sodium ((4-chloro-6-methoxypyrimidin-2-yl)carbamoyl)((2-
(isopropoxycarbonyl)phenoxy)sulfonyl)amide (5-16)
1
Yield 90%; m.p.: 164-166 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.96 (s,
6
1
H, CONH), 7.57 (d, J = 7.6 Hz, 1H, ArH), 7.52 – 7.44 (m, 2H, ArH), 7.24 – 7.19 (m,
1
H, ArH), 6.49 (s, 1H, Prim-H), 5.09 – 5.02 (m, 1H, OCH), 3.88 (s, 3H, OCH ), 1.29
3
1
3
(
d, J = 6.3 Hz, 6H, CH ); C NMR (101 MHz, DMSO-d ), δ 171.0, 165.9, 160.1,
3
6
1
59.0, 155.3, 150.6, 132.6, 130.3, 127.0, 124.8, 123.8, 99.2, 68.5, 54.6, 22.1; HRMS
+
+
(
ESI) m/z: calculated for C H ClN NaO S (M-Na +2H ) 445.0585, found 445.0587.
16 16 4 7
4
.3.17.
Sodium
((4-chloro-6-methoxypyrimidin-2-yl)carbamoyl)((2-
(phenoxycarbonyl)phenoxy)sulfonyl)amide (5-17)
1
Yield 97%; m.p.: 133-135 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.08 (s,
6
1
1
1
1
4
4
H, CONH), 7.79 (s, 1H, ArH), 7.61 (s, 1H, ArH), 7.54 – 7.25 (m, 7H, ArH), 6.50 (s,
1
3
H, Prim-H), 3.88 (s, 3H, OCH ); C NMR (101 MHz, DMSO-d ), δ 171.1, 165.1,
3
6
60.2, 159.0, 155.7, 151.3, 150.8, 133.7, 131.0, 129.7, 126.2, 126.0, 125.4, 124.5,
+
+
22.7, 99.3, 54.6; HRMS (ESI) m/z: C H ClN NaO S calculated for (M-Na +2H )
19
14
4
7
79.0421, found 479.0418.
.3.18. Sodium ((4-chloro-6-methoxypyrimidin-2-yl)carbamoyl)((2-nitrophenoxy)

ACCEPTED MANUSCRIPT
sulfonyl) amide (5-18)
1
Yield 75%; m.p.: 134-136 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.20 (s,
6
1
H, CONH), 7.84 (d, J = 7.4 Hz, 1H, ArH), 7.65 (d, J = 5.3 Hz, 2H, ArH), 7.33 (t, J =
13
1
2.6, 4.2 Hz, 1H, ArH), 6.52 (s, 1H, Prim-H), 3.89 (s, 3H, OCH ); C NMR (101
3
MHz, DMSO-d ), δ 170.4, 158.6, 155.2, 151.5, 144.7, 143.8, 134.0, 125.3, 124.9,
6
+
+
1
4
4
24.8, 103.4, 54.6; HRMS (ESI) m/z: calculated for C H ClN NaO S (M-Na +2H )
12 9 5 7
04.0063, found 404.0068.
.3.19. Sodium ((4-chloro-6-methoxypyrimidin-2-yl)carbamoyl)((4-(ethoxycarbonyl)
phenoxy)sulfonyl)amide (5-19)
Yield 73%; m.p.: 138-140 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.15 (s,
1
6
1
H, CONH), 7.91 (d, J = 8.6 Hz, 2H, ArH), 7.35 (d, J = 8.6 Hz, 2H, ArH), 6.49 (s, 1H,
Prim-H), 4.29 (t, J = 14.1, 7.0 Hz, 2H, OCH ), 3.87 (s, 3H, OCH ), 1.31 (t, J = 7.0 Hz,
2
3
1
3
3
1
H, OCH ); C NMR (101 MHz, DMSO-d ), δ 171.1, 165.7, 160.2, 158.8, 156.5,
3 6
56.0, 130.9, 126.2, 121.6, 99.4, 61.1, 54.7, 14.6; HRMS (ESI) m/z: calculated for
+
+
C H ClN NaO S (M-Na +2H ) 431.0423, found 431.0417.
15
14
4
7
4
.3.20. Sodium ((4-chloro-6-methoxypyrimidin-2-yl)carbamoyl)((4-nitrophenoxy)
sulfonyl)amide (5-20)
Yield 94%; m.p.: 157-159 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.23 (s,
1
6
1
H, CONH), 8.23 (d, J = 9.1 Hz, 2H, ArH), 7.47 (d, J = 9.0 Hz, 2H, ArH), 6.50 (s, 1H,
1
3
Prim-H), 3.87 (s, 3H, OCH ); C NMR (101 MHz, DMSO-d ), δ 171.0, 160.1, 158.8,
3
6
1
58.2, 155.6, 143.8, 125.6, 121.8, 99.5, 54.7; HRMS (ESI) m/z: calculated for
+
+
C H ClN NaO S (M-Na +2H ) 404.0063, found 404.0057.
12
9
5
7
4
.3.21. Sodium ((4-bromo-6-methoxypyrimidin-2-yl)carbamoyl)((2-(methoxycarbonyl)
phenoxy)sulfonyl)amide (5-21)
Yield 82%; m.p.: 220-222 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.99 (s,
1
6
1
H, CONH), 7.60 (d, J = 7.6 Hz, 1H, ArH), 7.54 – 7.44 (m, 2H, ArH), 7.23 (t, J = 7.0
13
Hz, 1H, ArH), 6.64 (s, 1H, Prim-H), 3.87 (s, 3H, OCH ), 3.76 (s, 3H, OCH ); C
3
3
NMR (101 MHz, DMSO-d ), δ 170.5, 166.8, 158.6, 155.5, 151.5, 150.6, 133.0, 130.5,
6
1
26.2, 125.0, 124.0, 103.2, 54.6, 52.5; HRMS (ESI) m/z: calculated for
+
+
C H BrN NaO S (M-Na +2H ) 460.9762, found 460.9759.
14
12
4
7
4
.3.22. Sodium ((4-bromo-6-methoxypyrimidin-2-yl)carbamoyl)((2-(ethoxycarbonyl)
phenoxy)sulfonyl)amide (5-22)
Yield 75%; m.p.: 199-201 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.02 (s,
1
6
1
H, CONH), 7.61 (d, J = 7.7 Hz, 1H, ArH), 7.55 – 7.44 (m, 2H, ArH), 7.23 (t, J = 7.9

ACCEPTED MANUSCRIPT
Hz, 1H, ArH), 6.65 (s, 1H, Prim-H), 4.24 (q, J = 7.1 Hz, 2H, OCH ), 3.88 (s, 3H,
2
13
OCH ), 1.28 (t, J = 7.1 Hz, 3H, CH CH ); C NMR (101 MHz, DMSO-d ), δ 170.5,
3
2
3
6
1
66.2, 158.6, 155.7, 151.5, 150.5, 132.9, 130.5, 126.4, 125.0, 123.9, 103.3, 61.2, 54.6,
+
+
1
4.4; HRMS (ESI) m/z: calculated for C H BrN NaO S (M-Na +2H ) 474.9918,
15 14 4 7
found 474.9919.
4
.3.23. Sodium ((4-bromo-6-methoxypyrimidin-2-yl)carbamoyl)((2-(propoxycarbonyl)
phenoxy)sulfonyl)amide (5-23)
Yield 81%; m.p.: 203-205 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.02 (s,
1
6
1
H, CONH), 7.61 (d, J = 7.8 Hz, 1H, ArH), 7.54 – 7.46 (m, 2H, ArH), 7.23 (t, J = 7.1
Hz, 1H, ArH), 6.65 (s, 1H, Prim-H), 4.14 (t, J = 6.6 Hz, 2H, OCH ), 3.87 (s, 3H,
2
13
OCH ), 1.69 (m, 2H, OCH CH ), 0.94 (t, J = 7.4 Hz, 3H, CH CH ); C NMR (101
3
2
2
2
3
MHz, DMSO-d ), δ 170.5, 166.3, 158.7, 155.4, 151.5, 150.6, 132.8, 130.4, 126.4,
6
1
24.9, 123.9, 103.2, 66.7, 54.6, 219, 10.9; HRMS (ESI) m/z: calculated for
+
+
C H BrN NaO S (M-Na +2H ) 489.0075, found 489.0067.
16
16
4
7
4
.3.24. sodium ((4-bromo-6-methoxypyrimidin-2-yl)carbamoyl)((2-((2-chloroethoxy)
carbonyl)phenoxy)sulfonyl)amide (5-24)
Yield 95%; m.p.: 178-180 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.04 (s,
1
6
1
H, CONH), 7.64 (d, J = 7.6 Hz, 1H, ArH), 7.55 (t, J = 7.1 Hz, 1H, ArH), 7.46 (d, J =
.0 Hz, 1H, ArH), 7.26 (t, J = 7.5 Hz, 1H, ArH), 6.65 (s, 1H, Prim-H), 4.45 (t, J = 5.7
8
13
Hz, 2H, OCH ), 3.93 (t, J = 5.7 Hz, 2H, CH Cl), 3.88 (s, 3H, OCH ); C NMR (101
2
2
3
MHz, DMSO-d ), δ 170.4, 165.8, 158.6, 155.4, 151.5, 150.7, 133.2, 130.5, 125.9,
6
1
25.1, 124.2, 103.2, 65.0, 54.6, 42.7; HRMS (ESI) m/z: calculated for
+
+
C H BrClN NaO S (M-Na +2H ) 508.9528, found 508.9524.
15
13
4
7
4
.3.25. Sodium ((4-bromo-6-methoxypyrimidin-2-yl)carbamoyl)((2-((2-iodoethoxy)
carbonyl)phenoxy)sulfonyl)amide (5-25)
Yield 77%; m.p.: 208-210 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.08 (s,
1
6
1
1
2
1
H, CONH), 7.46 (d, J = 7.5 Hz, 1H, ArH), 7.07 – 7.02 (m, 2H, ArH), 6.95 – 6.90 (m,
H, ArH), 6.64 (s, 1H, Prim-H), 4.24 (t, J = 5.6 Hz, 2H, OCH ), 3.90 (t, J = 5.6 Hz,
2
13
H, CH I), 3.87 (s, 3H, OCH ); C NMR (101 MHz, DMSO-d ), δ 170.4, 165.5,
2
3
6
58.6, 155.3, 151.5, 150.7, 133.2, 130.5, 125.9, 125.0, 124.1, 103.3, 65.4, 54.6, 2.8;
+
+
HRMS (ESI) m/z: calculated for C H BrIN NaO S (M-Na +2H ) 600.8885, found
1
5
13
4
7
6
00.8883.
.3.26. Sodium ((4-bromo-6-methoxypyrimidin-2-yl)carbamoyl)((2-(2-chloroethoxy)
4
phenoxy)sulfonyl)amide (5-26)

ACCEPTED MANUSCRIPT
1
Yield 81%; m.p.: 128-130 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.07 (s,
6
1
H, CONH), 7.47 (d, J = 7.4 Hz, 1H, ArH), 7.07 – 7.03 (m, 2H, ArH), 6.95 – 6.90 (m,
1
H, ArH), 6.63 (s, 1H, Prim-H), 4.24 (t, J = 5.6 Hz, 2H, OCH ), 3.92 – 3.89 (m, 2H,
2
13
CH Cl), 3.87 (s, 3H, OCH ); C NMR (101 MHz, DMSO-d ), δ 170.4, 158.7, 155.2,
2
3
6
1
51.5, 150.6, 142.5, 125.5, 123.6, 121.9, 116.6, 103.2, 70.2, 54.5, 43.3; HRMS (ESI)
+
+
m/z: calculated for C H BrClN NaO S (M-Na +2H ) 480.9579, found 480.9570.
1
4
13
4
6
4
.3.27. sodium ((4-bromo-6-methoxypyrimidin-2-yl)carbamoyl)((2-(2-iodoethoxy)
phenoxy)sulfonyl)amide (5-27)
Yield 78%; m.p.: 125-127 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.09 (s,
1
6
1
1
3
1
H，CONH), 7.45 (d, J = 7.5 Hz, 1H, ArH), 7.09 – 7.03 (m, 2H, ArH), 6.96 – 6.90 (m,
H, ArH), 6.65 (s, 1H, Prim-H), 4.25 (t, J = 6.9 Hz, 2H, OCH ), 3.88 (s, 3H, OCH ),
2
3
1
3
.47 (t, J = 6.9 Hz, 2H, CH I); C NMR (101 MHz, DMSO-d ), δ 170.5, 158.6, 155.4,
2
6
51.5, 150.4, 142.3, 125.7, 123.8, 121.9, 116.7, 103.3, 70.8, 54.6, 4.0; HRMS (ESI)
+
+
m/z: calculated for C H BrIN NaO S (M-Na +2H ) calcd. 572.8935, found
1
4
13
4
6
5
72.8930.
.3.28. Sodium ((4-bromo-6-methoxypyrimidin-2-yl)carbamoyl)((2-(isopropoxy-
4
carbonyl)phenoxy)sulfonyl)amide (5-28)
Yield 82%; m.p.: 162-164 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.97 (s,
1
6
1
H, CONH), 7.58 (d, J = 7.6 Hz, 1H, ArH), 7.53 – 7.44 (m, 2H, ArH), 7.22 (t, J = 7.3
Hz, 1H, ArH), 6.64 (s, 1H, Prim-H), 5.06 (dt, J = 12.5, 6.2 Hz, 1H, OCH), 3.88 (s, 3H,
1
3
OCH ), 1.30 (d, J = 6.3 Hz, 6H, CH ); C NMR (101 MHz, DMSO-d ), δ 170.4,
3
3
6
1
65.8, 158.7, 155.4, 151.5, 150.6, 132.7, 130.3, 126.9, 124.9, 123.8, 103.2, 68.4, 54.6,
+
+
2
2.1; HRMS (ESI) m/z: calculated for C H BrN NaO S (M-Na +2H ) 489.0075,
16 16 4 7
found 489.0085.
4
.3.29. Sodium ((4-bromo-6-methoxypyrimidin-2-yl)carbamoyl)((2-nitrophenoxy)
sulfonyl)amide (5-29)
Yield 85%; m.p.: 146-148 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.19 (s,
1
6
1
4
H, CONH), 7.84 (d, J = 7.6 Hz, 1H, ArH), 7.66 – 7.62 (m, 2H, ArH), 7.33 (t, J = 9.6,
1
3
.1 Hz, 1H, ArH), 6.66 (s, 1H, Prim-H), 3.87 (s, 3H, OCH ); C NMR (101 MHz,
3
DMSO-d ), δ 170.4, 158.6, 155.2, 151.5, 144.7, 143.8, 134.0, 125.3, 125.0, 124.8,
6
+
+
1
4
03.4, 54.6; HRMS (ESI) m/z: calculated for C H BrN NaO S (M-Na +2H )
12 9 5 7
47.9558, found 447.9529.

ACCEPTED MANUSCRIPT
4
.3.30. Sodium ((4-iodo-6-methoxypyrimidin-2-yl)carbamoyl)((2-(methoxycarbonyl)
phenoxy)sulfonyl)amide (5-30)
Yield 86%; m.p.: 206-208 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.95 (s,
1
6
1
H, CONH), 7.68 (d, J = 7.4 Hz, 1H, ArH), 7.61 – 7.52 (m, 2H, ArH), 7.30 (t, J = 7.0
13
Hz, 1H, ArH), 6.94 (s, 1H, Prim-H), 3.91 (s, 3H, OCH ), 3.84 (s, 3H, OCH ); C
3
3
NMR (101 MHz, DMSO-d ), δ 169.2, 166.8, 158.0, 155.6, 150.6, 133.0, 130.5, 129.5,
6
1
26.2, 125.0, 124.0, 110.6, 54.3, 52.5; HRMS (ESI) m/z: calculated for
+
C H IN NaO S (M+H ) 530.9443, found 530.9448.
14
12
4
7
4
.3.31. Sodium ((2-(ethoxycarbonyl)phenoxy)sulfonyl)((4-iodo-6-methoxypyrimidin-
2
-yl)carbamoyl)amide (5-31)
1
Yield 91%; m.p.: 196-198 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.84 (s,
6
1
H , CONH), 7.59 (d, J = 7.7 Hz, 1H, ArH), 7.53 – 7.44 (m, 2H, ArH), 7.22 (t, J = 7.4
Hz, 1H, ArH), 6.86 (s, 1H, Prim-H), 4.23 (q, J = 7.1 Hz, 2H, OCH ), 3.84 (s, 3H,
2
13
OCH ), 1.28 (t, J = 7.1 Hz, 3H, CH ); C NMR (101 MHz, DMSO-d ), δ 169.2,
3
3
6
1
66.3, 158.1, 155.4, 150.6, 132.8, 130.4, 129.5, 126.5, 124.9, 124.0, 110.5, 61.1, 54.3,
+
+
1
4.5; HRMS (ESI) m/z: calculated for C H IN NaO S (M-Na +2H ) 522.9779,
15 14 4 7
found 522.9773.
4
.3.32. Sodium ((4-iodo-6-methoxypyrimidin-2-yl)carbamoyl)((2-(propoxycarbonyl)
phenoxy)sulfonyl)amide (5-32)
Yield 89%; m.p.: 188-190 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.89 (s,
1
6
1
H，CONH), 7.61 (d, J = 7.4 Hz, 1H, ArH), 7.55 – 7.46 (m, 2H, ArH), 7.23 (t, J = 7.2
Hz, 1H, ArH), 6.88 (s, 1H, Prim-H), 4.14 (t, J = 6.6 Hz, 2H, OCH ), 3.84 (s, 3H,
2
13
OCH ), 1.69 (t, J = 14.2, 7.1 Hz, 2H, OCH CH ), 0.94 (t, J = 7.4 Hz, 3H, CH ); C
3
2
2
3
NMR (101 MHz, DMSO-d ), δ 169.2, 166.3, 158.0, 155.5, 150.6, 132.9, 130.4, 129.5,
6
1
26.4, 124.9, 123.9, 110.5, 66.7, 54.3, 21.9, 10.9; HRMS (ESI) m/z: calculated for
+
+
C H IN NaO S (M-Na +2H ) 536.9936, found 536.9928.
16
16
4
7
4
.3.33. Sodium ((2-((2-chloroethoxy)carbonyl)phenoxy)sulfonyl)((4-iodo-6-methoxy-
pyrimidin-2-yl)carbamoyl)amide (5-33)
Yield 77%; m.p.: 107-109 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.84 (s,
1
6
1
H, CONH), 7.64 (d, J = 4.2 Hz, 1H, ArH), 7.54 (t, J = 6.9 Hz, 1H, ArH), 7.46 (d, J =
.2 Hz, 1H, ArH), 7.25 (t, J = 7.5 Hz, 1H, ArH), 6.87 (s, 1H, Prim-H), 4.44 (t, J = 5.6
8
13
Hz, 2H, OCH ), 3.92 (t, J = 5.7 Hz, 2H, CH Cl), 3.84 (s, 3H, OCH ); C NMR (101
2
2
3
MHz, DMSO-d ), δ 169.2, 165.8, 158.1, 155.3, 150.7, 133.2, 130.5, 129.5, 125.9,
6

ACCEPTED MANUSCRIPT
1
25.0, 124.2, 110.5, 65.0, 54.2, 42.7; HRMS (ESI) m/z: calculated for
+
+
C H ClIN NaO S (M-Na +2H ) 556.9390, found 556.9382.
15
13
4
7
4
2
.3.34. Sodium ((2-(2-chloroethoxy)phenoxy)sulfonyl)((4-iodo-6-methoxypyrimidin-
-yl)carbamoyl)amide (5-34)
1
Yield 83%; m.p.: 128-130 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.98 (s,
6
1
1
2
1
H, CONH), 7.46 (d, J = 7.5 Hz, 1H, ArH), 7.07 – 7.03 (m, 2H, ArH), 6.96 – 6.90 (m,
H, ArH), 6.87 (s, 1H, Prim-H), 4.25 (t, J = 5.6 Hz, 2H, OCH ), 3.91 (t, J = 5.6 Hz,
2
1
3
H, CH Cl), 3.84 (s, 3H, OCH ); C NMR (101 MHz, DMSO-d ), δ 169.2, 158.1,
2
3
6
55.2, 150.6, 142.4, 129.5, 125.5, 123.6, 121.9, 116.6, 110.5, 70.2, 54.2, 43.3; HRMS
+
+
(
ESI) m/z: calculated for C H ClIN NaO S (M-Na +2H ) 528.9441, found
14 13 4 6
5
4
28.9435.
.3.35. Sodium ((4-iodo-6-methoxypyrimidin-2-yl)carbamoyl)((2-(2-iodoethoxy)
phenoxy)sulfonyl)amide (5-35)
Yield 88%; m.p.: 116-118 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.95 (s,
1
6
1
1
3
1
H, CONH), 7.44 (d, J = 8.3 Hz, 1H, ArH), 7.04 (m, 2H, ArH), 6.93 (d, J = 8.2 Hz,
H, ArH), 6.86 (s, 1H, Prim-H), 4.24 (t, J = 6.8 Hz, 2H, OCH ), 3.84 (s, 3H, OCH ),
2
3
1
3
.46 (t, J = 6.7 Hz, 2H, CH I); C NMR (101 MHz, DMSO-d ), δ 169.2, 158.0, 155.2,
2
6
50.4, 142.4, 129.5, 125.6, 123.8, 121.9, 116.7, 110.6, 70.8, 54.8, 4.1; HRMS (ESI)
+
+
m/z: calculated for C H I N O S (M-Na +2H ) 620.8797, found 620.8784.
1
4
14 2
4
6
4
.3.36.
Sodium
((4-iodo-6-methoxypyrimidin-2-yl)carbamoyl)((2-nitrophenoxy)
sulfonyl)amide (5-36)
1
Yield 78%; m.p.: 143-145°C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.06 (s,
6
1
H, CONH), 7.84 (d, J = 7.8 Hz, 1H, ArH), 7.65 (d, J = 4.9 Hz, 2H, ArH), 7.36 – 7.29
13
(
m, 1H, ArH), 6.89 (s, 1H, Prim-H), 3.84 (s, 3H, OCH ); C NMR (101 MHz,
3
DMSO-d ), δ 169.2, 158.0, 155.2, 143.8, 135.0, 134.0, 125.3, 124.9, 124.8, 110.7,
6
+
+
1
4
4
06.0, 54.3. HRMS (ESI) m/z: calculated for C H IN NaO S (M-Na +2H )
12 9 5 7
95.9419, found 495.9411.
.3.37. Sodium ((2-(ethoxycarbonyl)phenoxy)sulfonyl)((4-methoxy-6-methyl
pyrimidin-2-yl)carbamoyl)amide (5-37)
Yield 82%; m.p.: 223-225 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.46 (s,
1
6
1
H, CONH), 7.61 (d, J = 7.6 Hz, 1H, ArH), 7.55 – 7.44 (m, 2H, ArH), 7.23 (t, J = 7.3
Hz, 1H, ArH), 6.26 (s, 1H, Prim-H), 4.29 – 4.18 (m, 2H, OCH ), 3.84 (s, 3H, OCH ),
2
3
1
3
2
.25 (s, 3H, CH ), 1.28 (t, J = 7.1 Hz, 3H, CH CH ); C NMR (101 MHz, DMSO-d ),
3 2 3 6
δ 170.4, 168.6, 166.3, 158.8, 156.4, 150.5, 132.9, 130.5, 126.5, 125.0, 124.0, 99.5,

ACCEPTED MANUSCRIPT
+
+
6
4
4
1.2, 53.8, 23.9, 14.4; HRMS (ESI) m/z: calculated for C H N NaO S (M-Na +2H )
16 17 4 7
11.0969, found 411.0971.
.3.38. Sodium ((4-methoxy-6-methylpyrimidin-2-yl)carbamoyl)((2-(propoxycarbonyl)
phenoxy)sulfonyl)amide (5-38)
Yield 83%; m.p.: 200-202 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.37 (s,
1
6
1
H, CONH), 7.60 (d, J = 7.9 Hz, 1H, ArH), 7.50 (d, J = 6.2 Hz, 2H, ArH), 7.25 – 7.20
(
m, 1H, ArH), 6.25 (s, 1H, Prim-H), 4.13 (t, J = 6.7 Hz, 2H, OCH ), 3.84 (s, 3H,
2
OCH ), 2.24 (s, 3H, CH ), 1.68 (t, J = 14.2, 7.2 Hz, 2H, OCH CH ), 0.93 (t, J = 7.4
3
3
2
2
13
Hz, 3H, CH CH ); C NMR (101 MHz, DMSO-d ), δ 170.4, 168.4, 166.4, 158.9,
2
3
6
1
55.7, 150.7, 132.7, 130.4, 126.5, 124.8, 123.9, 99.3, 66.6, 53.6, 23.9, 21.9, 10.9;
+
+
HRMS (ESI) m/z: calculated for C H N NaO S (M-Na +2H ) 425.1126, found
1
7
19
4
7
4
25.1126.
.3.39. Sodium ((2-((2-iodoethoxy)carbonyl)phenoxy)sulfonyl)((4-methoxy-6-methyl
4
pyrimidin-2-yl)carbamoyl)amide (5-39)
Yield 90%; m.p.: 199-201 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.46 (s,
1
6
1
H, CONH), 7.66 (d, J = 7.7 Hz, 1H, ArH), 7.54 (t, J = 7.7 Hz, 1H, ArH), 7.46 (d, J =
.0 Hz, 1H, ArH), 7.26 (t, J = 7.5 Hz, 1H, ArH), 6.26 (s, 1H, Prim-H), 4.44 (t, J = 6.9
8
Hz, 2H, OCH ), 3.85 (s, 3H, OCH ), 3.47 (t, J = 6.9 Hz, 2H, CH I), 2.25 (s, 3H, CH );
2
3
2
3
1
3
C NMR (101 MHz, DMSO-d ), δ 170.4, 168.5, 165.5, 158.8, 156.3, 150.7, 133.3,
6
1
30.6, 125.8, 125.1, 124.1, 99.5, 65.5, 53.8, 23.9, 2.8; HRMS (ESI) m/z: calculated
+
+
for C H IN NaO S (M-Na +2H ) 536.9936, found 536.9932.
1
6
16
4
7
4
.3.40. Sodium ((2-(2-chloroethoxy)phenoxy)sulfonyl)((4-methoxy-6-methylpyrimidin
2-yl)carbamoyl)amide (5-40)
Yield 96%; m.p.: 121-123 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.58 (s,
-
1
6
1
H, CONH), 7.46 (d, J = 7.8 Hz, 1H, ArH), 7.05 (t, J = 5.4 Hz, 2H, ArH), 6.96 – 6.89
(
m, 1H, ArH), 6.26 (s, 1H, Prim-H), 4.24 (t, J = 5.6 Hz, 2H, OCH ), 3.91 (t, J = 5.6
2
1
3
Hz, 2H, CH Cl), 3.84 (s, 3H, OCH ), 2.24 (s, 3H, CH ); C NMR (101 MHz,
2
3
3
DMSO-d ), δ 170.4, 168.5, 158.9, 156.2, 150.6, 142.3, 125.6, 123.6, 121.8, 116.4,
6
9
9.5, 70.1, 53.7, 43.3, 23.9; HRMS (ESI) m/z: calculated for C H ClN NaO S
15 16 4 6
+
+
(
M-Na +2H ) calcd. 417.0631, found 417.0634.
4
.3.41. Sodium ((2-(2-iodoethoxy)phenoxy)sulfonyl)((4-methoxy-6-methylpyrimidin-
2
-yl)carbamoyl)amide (5-41)
1
Yield 87%; m.p.: 127-129 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.53 (s,
6
1
H, CONH), 7.44 (d, J = 8.4 Hz, 1H, ArH), 7.04 (d, J = 7.5 Hz, 2H, ArH), 6.95 – 6.89

ACCEPTED MANUSCRIPT
(
m, 1H, ArH), 6.24 (s, 1H, Prim-H), 4.23 (t, J = 6.9 Hz, 2H, OCH ), 3.84 (s, 3H,
2
13
OCH ), 3.46 (t, J = 6.9 Hz, 2H, CH I), 2.24 (s, 3H, CH ); C NMR (101 MHz,
3
2
3
DMSO-d ), δ 170.4, 168.5, 158.9, 155.8, 150.4, 142.4, 125.6, 123.8, 121.9, 116.6,
6
9
9.4, 70.7, 53.7, 623.9, 4.0. HRMS (ESI) m/z: calculated for C H IN NaO S
15 17 4 6
+
+
(
M-Na +2H ) 508.9987, found 508.9987.
.3.42. Sodium ((4-methoxy-6-methylpyrimidin-2-yl)carbamoyl)((2-((2,2,2-
trifluoroethoxy)carbonyl)phenoxy)sulfonyl)amide (5-42)
4
1
Yield 89%; m.p.: 121-122 °C; white solid; yield 89%;. H NMR (400 MHz,
DMSO-d ), δ 8.40 (s, 1H, CONH), 7.67 – 7.55 (m, 2H, ArH), 7.49 (d, J = 7.9 Hz, 1H,
6
ArH), 7.32 – 7.25 (m, 1H, ArH), 6.25 (s, 1H, Prim-H), 4.98 – 4.86 (m, 2H, CH CF ),
2
3
1
3
3
.84 (s, 3H, OCH ), 2.24 (s, 3H, CH ); C NMR (101 MHz, DMSO-d ), δ 170.4,
3 3 6
1
68.5, 164.5, 158.9, 156.0, 151.0, 143.3, 133.8, 125.2, 124.7, 124.4, 99.4, 61.0, 53.7,
+
+
2
3.8; HRMS (ESI) m/z: calculated for C H F N NaO S (M-Na +2H ) 465.0687,
1
6
14
3
4
7
found 465.0684.
4
.3.43. Sodium ((2-(isopropoxycarbonyl)phenoxy)sulfonyl)((4-methoxy-6-methyl
pyrimidin-2-yl)carbamoyl)amide (5-43)
1
Yield 72%; m.p.: 164-166 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.46 (s,
6
1
H, CONH), 7.65 (d, J = 9.1 Hz, 1H, ArH), 7.60 – 7.51 (m, 2H, ArH), 7.32 – 7.26 (m,
H, ArH), 6.32 (s, 1H, Prim-H), 5.14 (dt, J = 12.5, 6.3 Hz, 1H, OCH), 3.92 (s, 3H,
1
1
3
OCH ), 2.31 (s, 3H, CH ), 1.37 (d, J = 6.3 Hz, 6H, CH ); C NMR (101 MHz,
3
3
3
DMSO-d ), δ 170.4, 168.5, 165.9, 158.9, 155.9, 150.6, 132.6, 130.3, 126.9, 124.8,
6
1
23.8, 99.7, 68.5, 53.7, 23.9 22.1. HRMS (ESI) m/z: calculated for C H N NaO S
17 19 4 7
+
+
(
M-Na +2H ) 425.1126, found 425.1117.
4
.3.44. Sodium ((3-(ethoxycarbonyl)phenoxy)sulfonyl)((4-methoxy-6-methylpyrimidin
-2-yl)carbamoyl)amide (5-44)
1
Yield 83%; m.p.: 134-136 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.45 (s,
6
1
H, CONH), 7.80 (s, 1H, ArH), 7.73 (t, J = 7.0, 3.1 Hz, 1H, ArH), 7.53 – 7.45 (m, 2H,
ArH), 6.24 (s, 1H, Prim-H), 4.30 (q, J = 7.1 Hz, 2H, OCH ), 3.84 (s, 3H, OCH ), 2.23
2
3
1
3
(
s, 3H, CH ), 1.29 (t, J = 7.1 Hz, 3H, CH CH ); C NMR (101 MHz, DMSO-d ), δ
3 2 3 6
1
5
4
4
70.4, 168.5, 165.7, 158.9, 156.4, 152.7, 131.3, 130.0, 127.4, 125.8, 122.7, 99.6, 61.3,
+
+
3.7, 23.8, 14.6; HRMS (ESI) m/z: calculated for C H N NaO S (M-Na +2H )
1
6
17
4
7
11.0969, found 411.0971.
.3.45. Sodium ((4-methoxy-6-methylpyrimidin-2-yl)carbamoyl)((4-nitrophenoxy)
sulfonyl)amide (5-45)

ACCEPTED MANUSCRIPT
1
Yield 79%; m.p.: 148-150 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.60 (s,
6
1
H, CONH), 8.23 (d, J = 8.9 Hz, 2H, ArH), 7.48 (d, J = 8.9 Hz, 2H, ArH), 6.25 (s, 1H,
1
3
Prim-H), 3.83 (s, 3H, OCH ), 2.23 (s, 3H, CH ); C NMR (101 MHz, DMSO-d ), δ
3
3
6
1
70.4, 168.6, 158.8, 158.2, 156.3, 143.8, 125.6, 121.8, 99.6, 53.8, 23.8; HRMS (ESI)
+
+
m/z: calculated for C H N NaO S (M-Na +2H ) 384.0609, found 384.0613.
1
3
12
5
7
4
.3.46.
phenoxy)sulfonyl)amide (5-46)
Yield 87%; m.p.: 122-124 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.37 (s,
Sodium
((4,6-dimethylpyrimidin-2-yl)carbamoyl)((2-(ethoxycarbonyl)
1
6
1
H, CONH), 7.61 (d, J = 6.2 Hz, 1H, ArH), 7.54 – 7.45 (m, 2H, ArH), 7.23 (t, J = 7.3
Hz, 1H, ArH), 6.72 (s, 1H, Prim-H), 4.23 (q, J = 7.1 Hz, 2H, OCH ), 2.28 (s, 6H,
2
13
CH ), 1.28 (t, J = 7.1 Hz, 3H, CH ); C NMR (101 MHz, DMSO-d ), δ 167.7, 166.3,
3
3
6
1
59.0, 156.5, 150.5, 132.8, 130.4, 126.5, 125.0, 124.0, 114.0, 61.2, 23.9, 14.4; HRMS
+
+
(
ESI) m/z: calculated for C H N NaO S (M-Na +2H ) 395.1020, found 395.1026.
16 17 4 6
4
.3.47.
Sodium
((4,6-dimethylpyrimidin-2-yl)carbamoyl)((2-(propoxycarbonyl)
phenoxy)sulfonyl)amide (5-47)
Yield 92%; m.p.: 208-210 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.31 (s,
1
6
1
H, CONH), 7.60 (d, J = 8.1 Hz, 1H, ArH), 7.49 (d, J = 2.5 Hz, 2H, ArH), 7.25 – 7.19
(
m, 1H, ArH), 6.71 (s, 1H, Prim-H), 4.13 (t, J = 6.7 Hz, 2H, OCH ), 2.28 (s, 6H, CH ),
2
3
1
3
1
.68 (dd, J = 14.3, 7.1 Hz, 2H, OCH CH ), 0.94 (t, J = 7.4 Hz, 3H, CH CH ); C
2 2 2 3
NMR (101 MHz, DMSO-d ), δ 167.7, 166.4, 159.0, 156.4, 150.6, 132.8, 130.4, 126.4,
6
1
25.0, 124.0, 114.0, 66.7, 23.9, 21.9, 10.9; HRMS (ESI) m/z: calculated for
+
+
C H N NaO S (M-Na +2H ) 409.1177, found 409.1183.
17
19
4
6
4
.3.48.
pyrimidin-2-yl)carbamoyl)amide (5-48)
Yield 85%; m.p.: 190-192 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.41 (s,
Sodium((2-((2-chloroethoxy)carbonyl)phenoxy)sulfonyl)((4,6-dimethyl
1
6
1
H, CONH), 7.64 (d, J = 7.7 Hz, 1H, ArH), 7.55 (t, J = 7.8 Hz, 1H, ArH), 7.47 (d, J =
.2 Hz, 1H, ArH), 7.26 (t, J = 7.4 Hz, 1H, ArH), 6.73 (s, 1H, Prim-H), 4.44 (t, J = 5.7
8
1
3
Hz, 2H, OCH ), 3.92 (t, J = 5.7 Hz, 2H, CH Cl), 2.29 (s, 6H, CH ); C NMR (101
2
2
3
MHz, DMSO-d ), δ 167.7, 165.8, 158.9, 156.6, 150.6, 133.3, 130.5, 125.8, 125.2,
6
1
24.2, 114.2, 65.0, 42.6, 23.8; HRMS (ESI) m/z: calculated for C H IN O S
16 17 4 6
+
+
(
M-Na +2H ) 429.0631, found 429.0633.
4
.3.49. Sodium ((4,6-dimethylpyrimidin-2-yl)carbamoyl)((2-((2-iodoethoxy)carbonyl)
phenoxy)sulfonyl)amide (5-49)
Yield 93%; m.p.: 200-202 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.39 (s,
1
6

ACCEPTED MANUSCRIPT
1
H, CONH), 7.66 (d, J = 8.8 Hz, 1H, ArH), 7.54 (t, J = 7.0 Hz, 1H, ArH), 7.46 (d, J =
.0 Hz, 1H, ArH), 7.26 (t, J = 7.4 Hz, 1H, ArH), 6.73 (s, 1H, Prim-H), 4.44 (t, J = 6.9
8
1
3
Hz, 2H, OCH ), 3.47 (t, J = 6.9 Hz, 2H, CH I), 2.29 (s, 6H, CH ); C NMR (101
2
2
3
MHz, DMSO-d ), δ 167.8, 165.5, 159.0, 156.6, 150.7, 133.3, 130.6, 125.8, 125.2,
6
1
24.2, 114.1, 65.5, 23.9, 2.8; HRMS (ESI) m/z: calculated for C H IN NaO S
16 17 4 6
+
+
(
M-Na +2H ) 520.9987, found 520.9986.
4
.3.50. Sodium ((2-(2-chloroethoxy)phenoxy)sulfonyl)((4,6-dimethylpyrimidin-2-yl)
carbamoyl)amide (5-50)
Yield 97%; m.p.: 119-121 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.57 (s,
1
6
1
H, CONH), 7.46 (d, J = 8.8 Hz, 1H, ArH), 7.05 (t, J = 6.7 Hz, 2H, ArH), 6.95 – 6.89
(
m, 1H, ArH), 6.73 (s, 1H, Prim-H), 4.23 (t, J = 5.7 Hz, 2H, OCH ), 3.90 (t, J = 5.7
2
1
3
Hz, 2H, CH Cl), 2.28 (s, 6H, CH ); C NMR (101 MHz, DMSO-d ), δ 167.9, 159.0,
2
3
6
1
56.6, 150.7, 142.1, 125.8, 123.7, 121.8, 116.3, 114.1, 70.1, 43.3, 23.9; HRMS (ESI)
+
+
m/z: calculated for C H ClN NaO S (M-Na +2H ) 401.0681, found 401.0686.
1
5
16
4
5
4
.3.51. Sodium ((4,6-dimethylpyrimidin-2-yl)carbamoyl)((2-(2-iodoethoxy)phenoxy)
sulfonyl)amide (5-51)
Yield 78%; m.p.: 118-120 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.56 (s,
1
6
1
1
2
1
H, CONH), 7.44 (d, J = 8.4 Hz, 1H, ArH), 7.08 – 7.01 (m, 2H, ArH), 6.95 – 6.88 (m,
H, ArH), 6.72 (s, 1H, Prim-H), 4.23 (t, J = 6.9 Hz, 2H, OCH ), 3.46 (t, J = 6.9 Hz,
2
13
H, CH I), 2.29 (s, 6H, CH ); C NMR (101 MHz, DMSO-d ), δ 167.7, 159.0, 156.2,
2
3
6
50.5, 142.3, 125.7, 123.8, 121.9, 116. 5, 114.0, 70.6, 23.9, 4.0; HRMS (ESI) m/z:
+
+
calculated for C H IN NaO S (M-Na +2H ) 493.0038, found 493.0038.
1
5
16
4
5
4
.3.52. Sodium ((4,6-dimethylpyrimidin-2-yl)carbamoyl)((2-(isopropoxycarbonyl)
phenoxy)sulfonyl)amide (5-52)
Yield 90%; m.p.: 165-167 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.42 (s,
1
6
1
H, CONH), 7.65 (d, J = 7.4 Hz, 1H, ArH), 7.56 (q, J = 8.2 Hz, 2H, ArH), 7.32 – 7.27
(m, 1H, ArH), 6.80 (s, 1H, Prim-H), 5.14 (dt, J = 12.5, 6.2 Hz, 1H, OCH), 2.36 (s, 6H,
1
3
CH ), 1.37 (d, J = 6.3 Hz, 6H, CH ); C NMR (101 MHz, DMSO-d ), δ 167.7, 165.9,
3
3
6
1
59.1, 156.47, 150.6, 132.7, 130.3, 127.0, 124.9, 123.9, 114.0, 68.6, 23.9, 22.0;
+
+
HRMS (ESI) m/z: calculated for C H N NaO S (M-Na +2H ) 409.1177, found
1
7
19
4
6
4
09.1179.
4
.3.53.
Sodium
((4,6-dimethylpyrimidin-2-yl)carbamoyl)((2-(phenoxycarbonyl)
phenoxy)sulfonyl)amide (5-53)

ACCEPTED MANUSCRIPT
1
Yield 79%; m.p.: 137-139 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.37 (s,
6
1
8
1
1
1
4
4
H, CONH), 7.78 (d, J = 7.6 Hz, 1H, ArH), 7.60 (t, J = 7.8 Hz, 1H, ArH), 7.49 (d, J =
.3 Hz, 1H, ArH), 7.43 (t, J = 7.7 Hz, 2H, ArH), 7.39 – 7.25 (m, 4H, ArH), 6.72 (s,
1
3
H, Prim-H), 2.28 (s, 6H, CH ); C NMR (101 MHz, DMSO-d ), δ 167.7, 165.2,
3
6
59.1, 156.7, 151.3, 150.7, 133.7, 131.0, 129.8, 126.3, 126.1, 125.5, 124.6, 122.6,
+
+
14.1, 23.9; HRMS (ESI) m/z: calculated for C H N NaO S (M-Na +2H )
2
0
17
4
6
43.1020, found 443.1026.
.3.54. Sodium ((4,6-dimethylpyrimidin-2-yl)carbamoyl)((3-(ethoxycarbonyl)phenoxy)
sulfonyl)amide (5-54)
Yield 80%; m.p.: 126-128 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.44 (s,
1
6
1
2
1
1
H, CONH), 7.80 (m, 1H, ArH), 7.72 (d, J = 8.1 Hz, 1H, ArH), 7.47 (d, J = 6.3 Hz,
H, ArH), 6.72 (s, 1H, Prim-H), 4.29 (q, J = 7.1 Hz, 2H, CH ), 2.27 (s, 6H, CH ),
2
3
1
3
.29 (t, J = 7.1 Hz, 3H, CH CH ); C NMR (101 MHz, DMSO-d ), δ 167.7, 165.7,
2
3
6
59.0, 156.8, 152.7, 131.3, 130.0, 127.2, 125.8, 122.7, 114.1, 61.4, 23.8, 14.5; HRMS
+
+
(
ESI) m/z: calculated for C H N NaO S (M-Na +2H ) 395.1020, found 395.1015.
16 17 4 6
4
.3.55.
Sodium
((4,6-dimethylpyrimidin-2-yl)carbamoyl)((4-nitrophenoxy)
sulfonyl)amide (5-55)
1
Yield 89%; m.p.: 159-161 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.52 (s,
6
1
H, CONH), 8.22 (d, J = 9.2 Hz, 2H, ArH), 7.48 (d, J = 9.2 Hz, 2H, ArH), 6.72 (s, 1H,
13
Prim-H), 2.27 (s, 6H, CH ); C NMR (101 MHz, DMSO-d ), δ 167.7, 159.0, 158.3,
3
6
1
56.6, 143.9, 125.6, 121.9, 114.2, 23.8; HRMS (ESI) m/z: calculated for
+
+
C H N O S (M-Na +2H ) 368.0660, found 368.0657.
13
13
5
6
4
.3.56. Sodium ((4-chloro-6-methylpyrimidin-2-yl)carbamoyl)((2-(methoxycarbonyl)
phenoxy)sulfonyl)amide (5-56)
Yield 86%; m.p.: 130-131 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.99 (s,
1
6
1
H, CONH), 7.61 (d, J = 6.2 Hz, 1H, ArH), 7.50 (t, J = 13.5, 6.8 Hz, 2H, ArH), 7.23 (t,
J = 8.1 Hz, 1H, ArH), 6.98 (s, 1H, Prim-H), 3.77 (s, 3H, OCH ), 2.33 (s, 3H, CH );
3
3
1
3
C NMR (101 MHz, DMSO-d ), δ 170.5, 166.8, 160.3, 159.2, 156.0, 150.5, 133.0,
6
1
30.5, 126.2, 125.1, 124.1, 113.6, 52.5, 23.8; HRMS (ESI) m/z: calculated for
+
+
C H ClN NaO S (M-Na +2H ) 401.0318, found 401.0315.
14
12
4
5
4
.3.57. Sodium ((4-chloro-6-methylpyrimidin-2-yl)carbamoyl)((2-(ethoxycarbonyl)
phenoxy)sulfonyl)amide (5-57)
Yield 86%; m.p.: 120-122 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.95 (s,
1
6
1
H, CONH), 7.60 (d, J = 7.8 Hz, 1H, ArH), 7.53 – 7.45 (m, 2H, ArH), 7.25 – 7.20 (m,

ACCEPTED MANUSCRIPT
1
1
1
H, ArH), 6.98 (s, 1H, Prim-H), 4.23 (q, J = 7.1 Hz, 2H, OCH ), 2.33 (s, 3H, CH ),
2
3
1
3
.28 (t, J = 7.1 Hz, 3H, CH CH ); C NMR (101 MHz, DMSO-d ), δ 170.5, 166.3,
2
3
6
60.2, 159.3, 155.7, 150.6, 132.8, 130.4, 126.5, 125.0, 124.0, 113.5, 61.1, 23.8, 14.4;
+
+
HRMS (ESI) m/z: calculated for C H ClN NaO S (M-Na +2H ) 415.0474, found
1
5
14
4
6
4
15.0476.
.3.58. Sodium ((4-chloro-6-methylpyrimidin-2-yl)carbamoyl)((2-(propoxycarbonyl)
4
phenoxy)sulfonyl)amide (5-58)
1
Yield 80%; m.p.: 108-110 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.97 (d,
6
J = 6.9 Hz, 1H, CONH), 7.60 (d, J = 7.9 Hz, 1H, ArH), 7.50 (d, J = 5.7 Hz, 2H, ArH),
7
.22 (t, J = 6.8 Hz, 1H, ArH), 6.98 (s, 1H, Prim-H), 4.13 (t, J = 6.7 Hz, 2H, OCH2),
.32 (s, 3H, CH ), 1.68 (t, J = 14.2, 7.0 Hz, 2H, CH CH ), 0.94 (t, J = 7.4 Hz, 3H,
2
3
2
2
13
CH CH ); C NMR (101 MHz, DMSO-d ), δ 170.5, 166.3, 160.3, 159.2, 156.0, 150.6,
2
3
6
1
32.9, 130.5, 126.4, 125.0, 123.9, 113.6, 66.7, 23.8, 21.9, 10.9; HRMS (ESI) m/z:
+
+
calculated for C H ClN NaO S (M-Na +2H ) 429.0631, found 429.0632.
1
6
16
4
6
4
.3.59. Sodium ((4-chloro-6-methylpyrimidin-2-yl)carbamoyl)((2-((2-chloroethoxy)
carbonyl)phenoxy)sulfonyl)amide (5-59)
Yield 75%; m.p.: 143-145 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.98 (s,
1
6
1
H，CONH), 7.63 (d, J = 9.4 Hz, 1H, ArH), 7.57 – 7.51 (m, 1H, ArH), 7.46 (d, J = 8.2
Hz, 1H, ArH), 7.26 (t, J = 6.9 Hz, 1H, ArH), 6.98 (s, 1H, Prim-H), 4.45 (t, J = 5.7 Hz,
13
2
H, OCH ), 3.95 – 3.90 (m, 2H, CH Cl), 2.33 (s, 3H, OCH ); C NMR (101 MHz,
2 2 3
DMSO-d ), δ 170.5, 165.8, 160.3, 159.2, 155.8, 150.7, 133.3, 130.5, 125.9, 125.1,
6
1
24.2, 113.6, 65.0, 42.7, 23.9; HRMS (ESI) m/z: calculated for C H Cl N NaO S
15 13 2 4 6
+
+
(
M-Na +2H ) 449.0084, found 449.0087.
4
.3.60. Sodium ((4-chloro-6-methylpyrimidin-2-yl)carbamoyl)((2-((2-iodoethoxy)
carbonyl)phenoxy)sulfonyl)amide (5-60)
Yield 83%; m.p.: 199-201 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.98 (s,
1
6
1
H, CONH), 7.65 (d, J = 6.4 Hz, 1H, ArH), 7.54 (t, J = 7.7 Hz, 1H, ArH), 7.46 (d, J =
.1 Hz, 1H, ArH), 7.26 (t, J = 7.5 Hz, 1H, ArH), 6.99 (s, 1H, Prim-H), 4.44 (t, J = 6.9
8
1
3
Hz, 2H, OCH ), 3.47 (t, J = 6.9 Hz, 2H, CH I), 2.33 (s, 3H, CH ); C NMR (101
2
2
3
MHz, DMSO-d ), δ 170.5, 165.5, 160.3, 159.2, 155.8, 150.7, 133.2, 130.6, 125.9,
6
1
25.1, 124.2, 113.6, 65.4, 23.9, 2.8; HRMS (ESI) m/z: calculated for
+
+
C H ClIN NaO S (M-Na +2H ) 540.9441, found 540.9437.
15
13
4
6
4
.3.61. Sodium ((4-chloro-6-methylpyrimidin-2-yl)carbamoyl)((2-(2-chloroethoxy)

ACCEPTED MANUSCRIPT
phenoxy)sulfonyl)amide (5-61)
1
Yield 78%; m.p.: 119-121 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.09 (s,
6
1
1
H, CONH), 7.48 (d, J = 7.9 Hz, 1H, ArH), 7.06 (d, J = 5.9 Hz, 2H, ArH), 6.99 (m,
H, ArH), 6.95(s, 1H, Prim-H), 4.25 (t, J = 5.5 Hz, 2H, OCH ), 3.91 (t, J = 5.5 Hz, 2H,
2
13
CH Cl), 2.33 (s, 3H, CH ); C NMR (101 MHz, DMSO), δ 170.5, 160.3, 159.2,
2
3
1
55.8, 150.6, 142.3, 125.7, 123.7, 121.8, 116.4, 113.6, 70.1, 43.3, 23.8; HRMS (ESI)
+
+
m/z: calculated for C H Cl N NaO S (M-Na +2H ) 421.0135, found 421.0136.
1
4
13
2
4
5
4
.3.62. Sodium ((4-chloro-6-methylpyrimidin-2-yl)carbamoyl)((2-(2-iodoethoxy)
phenoxy)sulfonyl)amide (5-62)
Yield 93%; m.p.: 106-108 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.07 (s,
1
6
1
H, CONH), 7.45 (d, J = 8.0 Hz, 1H, ArH), 7.04 (s, 2H, ArH), 6.98 (s, 1H, ArH), 6.91
(
s, 1H, Prim-H), 4.24 (t, J = 6.7 Hz, 2H, OCH ), 3.46 (t, J = 6.7 Hz, 2H, CH I), 2.33
2
2
1
3
(
s, 3H, CH ); C NMR (101 MHz, DMSO-d ), δ 170.4, 160.3, 159.2, 155.5, 150.4,
3
6
1
42.3, 125.7, 123.8, 121.9, 116.6, 113.5, 70.7, 23.9, 4.0; HRMS (ESI) m/z: calculated
+
+
for C H ClIN NaO S (M-Na +2H ) 512.9491, found 512.9491.
1
4
13
4
5
4
.3.63.
Sodium
((4-chloro-6-methylpyrimidin-2-yl)carbamoyl)((2-(isopropoxy
carbonyl)phenoxy)sulfonyl)amide (5-63)
Yield 95%; m.p.: 160-162 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.99 (s,
1
6
1
H, CONH), 7.59 (d, J = 7.8 Hz, 1H, ArH), 7.49 (q, J = 6.6 Hz, 2H, ArH), 7.25 – 7.20
(m, 1H, ArH), 6.99 (s, 1H, Prim-H), 5.07 (dt, J = 12.5, 6.3 Hz, 1H, OCH), 2.33 (s, 3H,
1
3
CH ), 1.30 (d, J = 6.3 Hz, 6H, CH ); C NMR (101 MHz, DMSO-d ), δ 170.5, 165.8,
3
3
6
1
60.3, 159.2, 156.0, 150.6, 132.7, 130.4, 126.9, 125.0, 123.9, 113.5, 68.6, 23.8, 22.1;
+
+
HRMS (ESI) m/z: calculated for C H ClN NaO S (M-Na +2H ) 429.0631, found
1
6
16
4
6
4
29.0633.
.3.64. Sodium ((4-chloro-6-methylpyrimidin-2-yl)carbamoyl)((2-(phenoxycarbonyl)
4
phenoxy)sulfonyl)amide (5-64)
Yield 82%; m.p.: 125-127 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.03 (s,
1
6
1
H, CONH), 7.78 (d, J = 7.2 Hz, 1H, ArH), 7.61 (t, J = 7.8 Hz, 1H, ArH), 7.52 – 7.40
13
(
m, 3H, ArH), 7.39 – 7.25 (m, 4H, ArH), 6.99 (s, 1H, Prim-H), 2.32 (s, 3H, CH ); C
3
NMR (101 MHz, DMSO-d ), δ 167.7, 165.2, 159.1, 156.7, 151.3, 150.7, 133.7, 131.0,
6
1
29.8, 126.3, 126.1, 125.5, 124.6, 122.6, 114.1, 23.9; HRMS (ESI) m/z: calculated for
+
+
C H ClN NaO S (M-Na +2H ) 463.0474, found 463.0682.
19
14
4
6
4
.3.65.
Sodium
((4-chloro-6-methylpyrimidin-2-yl)carbamoyl)((2-nitrophenoxy)
sulfonyl)amide (5-65)

ACCEPTED MANUSCRIPT
1
Yield 77%; m.p.: 140-142 °C; White solid; H NMR (400 MHz, DMSO-d ), δ 9.16 (s,
6
1
H, CONH), 7.84 (d, J = 7.9 Hz, 1H, ArH), 7.65 (d, J = 3.9 Hz, 2H, ArH), 7.37 – 7.30
13
(
m, 1H, ArH), 7.00 (s, 1H, Prim-H), 2.33 (s, 3H, CH ); C NMR (101 MHz,
3
DMSO-d ), δ 170.4, 160.2, 159.2, 155.4, 144.7, 143.8, 134.0, 125.3, 124.9, 124.9,
6
+
+
1
3
4
13.6, 23.8; HRMS (ESI) m/z: calculated for C H ClN NaO S (M-Na +2H )
12 9 5 6
88.0114, found 388.0116.
.3.66. Sodium ((4-chloro-6-methylpyrimidin-2-yl)carbamoyl)((3-(ethoxycarbonyl)
phenoxy)sulfonyl)amide (5-66)
Yield 81%; m.p.: 137-139 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.12 (s,
1
6
1
6
7
1
H, CONH), 7.79 (m, 1H, ArH), 7.73 (d, J = 6.7 Hz, 1H, ArH), 7.48 (m, 2H, ArH),
.98 (s, 1H, Prim-H), 4.29 (q, J = 7.0 Hz, 2H, OCH ), 2.32 (s, 3H, CH ), 1.28 (t, J =
2
3
1
3
.1 Hz, 3H, CH CH ); C NMR (101 MHz, DMSO-d ), δ 170.5, 165.7, 160.3, 159.2,
2
3
6
56.3, 152.6, 131.4, 130.0, 127.2, 125.9, 122.8, 113.6, 61.4, 23.8, 14.5; HRMS (ESI)
+
+
m/z: calculated for C H ClN NaO S (M-Na +2H ) 415.0474, found 415.0476.
1
5
14
4
6
4
.3.67. Sodium ((4-chloro-6-methylpyrimidin-2-yl)carbamoyl)((4-(ethoxycarbonyl)
phenoxy)sulfonyl)amide (5-67)
Yield 92%; m.p.: 135-137 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 9.10 (s,
1
6
1
H, CONH), 7.91 (d, J = 8.8 Hz, 2H, ArH), 7.35 (d, J = 8.8 Hz, 2H, ArH), 6.97 (s, 1H,
Prim-H), 4.29 (q, J = 7.1 Hz, 2H, OCH ), 2.31 (s , 3H, CH ), 1.30 (t, J = 7.1 Hz, 3H,
2
3
13
CH CH ); C NMR (101 MHz, DMSO-d ), δ 166.0, 160.2, 159.3, 156.8, 155.7, 138.2,
2
3
6
1
31.0, 126.1, 121.6, 113.5, 61.1, 23.8 14.5; HRMS (ESI) m/z: calculated for
+
+
C H ClN NaO S (M-Na +2H ) 415.0474, found 415.0475.
15
14
4
6
4
.3.68.
Sodium
((4-chloro-6-methylpyrimidin-2-yl)carbamoyl)((4-nitrophenoxy)
sulfonyl)amide (5-68)
1
Yield 88%; m.p.: 158-160 °C; white solid; H NMR (400 MHz, DMSO-d ), δ 8.37 (s,
6
1
H, CONH), 8.26 (d, J = 9.1 Hz, 2H, ArH), 7.48 (d, J = 9.2 Hz, 2H, ArH), 6.06 (s, 1H,
13
Prim-H), 2.25 (s, 3H, CH ); C NMR (101 MHz, DMSO-d ), δ 170.5, 160.2, 159.2,
3
6
1
58.2, 155.8, 143.8, 125.6, 121.8, 113.7, 23.8; HRMS (ESI) m/z: calculated for
+
+
C H ClN NaO S (M-Na +2H ) 388.0114, found 388.0110.
12
9
5
6
4
.3.69.
4
.4 Enzyme assay of fungal AHAS inhibition
Catalytic subunit of C. albicans AHAS was expressed and purified as described