(Chlorosulfonyl)benzenesulfonyl FluoridesâVersatile Building Blocks

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(Chlorosulfonyl)benzenesulfonyl fluorides – versatile building blocks for combinatorial chemistry. Design, synthesis and evaluation of a covalent inhibitor library Kateryna Tolmachova, Yurii S. Moroz, Angelika Konovets, Maxim Platonov, Oleksandr Vasylchenko, Petro Borysko, Sergey Zozulya, Anastasia Gryniukova, Andrey V. Bogolubsky, Sergey Pipko, Pavel K. Mykhailiuk, Volodymyr Brovarets, and Oleksandr O. Grygorenko ACS Comb. Sci., Just Accepted Manuscript • DOI: 10.1021/acscombsci.8b00130 • Publication Date (Web): 24 Oct 2018 Downloaded from http://pubs.acs.org on October 27, 2018

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ACS Combinatorial Science

(Chlorosulfonyl)benzenesulfonyl fluorides – versatile building blocks for combinatorial chemistry. Design, synthesis and evaluation of a covalent inhibitor library Kateryna A. Tolmachova,a,b Yurii S. Moroz,a,e Angelika Konovets,a,c Maxim O. Platonov,c Oleksandr V. Vasylchenko,c Petro Borysko,d Sergey Zozulya,d Anastasia Gryniukova,d Andrey V. Bogolubsky,c Sergey Pipko,c Pavel K. Mykhailiuk,a Volodymyr S. Brovarets,b Oleksandr O. Grygorenkoa,c* a National

Taras Shevchenko University of Kyiv, Volodymyrska Street 60, Kyiv 01601, Ukraine

b Institute

of Bioorganic Chemistry & Petrochemistry, NAS of Ukraine Murmanska Street 1, Kyiv 02660, Ukraine

c Enamine

Ltd., Chervonotkatska Street 78, Kyiv 02094, Ukraine, www.enamine.net

d Bienta/Enamine

Ltd., Chervonotkatska Street 78, Kyiv 02094, Ukraine, www.bienta.net

ACS Paragon Plus Environment

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ACS Combinatorial Science 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

eChemspace,

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Ilukstes iela 38‐5, Riga, LV‐1082, Latvia, www.chem-space.com

E-mail: [email protected]

Keywords: Sulfonyl halides; Parallel synthesis; Chemoselectivity; Covalent fragments; Sulfonamides; Serine protease inhibitors

O Cl S O

1 2

R R NH

R1 2 O N R S O O S F O

O S F O R1 = hetaryl, EWGCH(R3) R2, R3 = H, alkyl

Abstract.

Multigram

synthesis

of

O N S S O N N O S F O Trypsin inhibitor IC50 = 84 M

(chlorosulfonyl)benzenesulfonyl

fluorides

is

described. Selective modification of these building blocks at the sulfonyl chloride function under parallel synthesis conditions is achieved. It is shown that the reaction scope includes the use of (hetero)aromatic and electron-poor aliphatic amines (e.g. amino nitriles). Utility of the method is demonstrated by preparation of the sulfonyl


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fluoride library for potential use as covalent fragments, which is demonstrated by a combination of in silico and in vitro screening against trypsin as a model enzyme. As a result, several inhibitors were identified with activity on the par to that of the known inhibitor.


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Introduction.

Sulfonyl fluorides have gained much interest in recent years primarily due to their reasonable reactivity towards nucleophilic attack. In organic synthesis, the sulfur (VI) – fluoride exchange (SufFEx) reaction was named as “another good reaction for click chemistry” due to its high efficiency and tolerance to a range of functional groups.1 In medicinal chemistry, sulfonyl fluorides have been considered as “privileged warheads” for design of covalent modifiers, which is again due to their controllable reactivity in the biological systems.2 The best known representatives of this class are benzyl sulfonyl fluoride (PMSF) and 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF), widely used in biochemical studies to inhibit serine proteases in an irreversible manner (Figure 1).3 These compounds are also examples of covalent fragments, which have attracted much attention in recent years.4

SO2F

Cl

SO2F +

H3N

PMSF

AEBSF

Figure 1. Known sulfonyl fluorides – covalent serine protease inhibitors


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Either the SufFEx methodology or a design of covalent inhibitors can benefit from functionalized sulfonyl fluorides, especially if the use of a parallel synthesis methodology is intended. Recent examples of building blocks illustrating this idea include sulfonyl fluorides bearing the moieties of heteroaliphatic amines,5 chalcones,6 aldehydes,7 carboxylic acids,8 organoboronates,9 as well as benzyl10,11 aryl,9 or vinyl12 bromides. Meanwhile, it is widely accepted that sulfonyl fluorides and sulfonyl chlorides differ significantly in their reactivity.13 Therefore, chlorosulfonyl-substituted sulfonyl fluorides might be envisaged as useful building blocks for the parallel synthesis of libraries of sulfonyl fluorides bearing a sulfonamide moiety.

Although examples of such building blocks are known in the literature for more than nine decades (compounds 1–7, Figure 2),14–17 only a few isolated examples that demonstrate their selective modification at the sulfonyl chloride moiety have been reported to date.15–20 In this work, we describe synthesis of all three isomeric (chlorosulfonyl)benzenesulfonyl fluorides 1–3, as well as demonstrate their utility for the preparation of a library of sulfonamide-containing sulfonyl fluorides 8 (Scheme 1). In


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addition to that, their potential for the covalent inhibition of serine proteases was evaluated using the classical target of this class – trypsin – as a model enzyme.

SO2F

ClO2S

SO2Cl

X 2, X = H 4, X = Me 5, X = OH

1

SO2F

SO2F ClO2S 3

SO2F

SO2F

ClO2S

ClO2S 6

7

Figure 2. Known examples of chlorosulfonyl-substituted sulfonyl fluorides

SO2F ClO2S 1, o-isomer 2, m-isomer 3, p-isomer

R1

H N 2 R R2 O 9 N S R1 O

SO2F

8

Scheme 1.

Results and discussion.

It should be noted that although all three sulfonyl fluorides 1–3 have been mentioned in the literature, their syntheses were not described in the corresponding papers; preparation of the compound 2 was mentioned only in a patent.21 We have prepared all


6

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three building blocks 1–3 using a modification of the above-mentioned method (Scheme 2).21 The synthesis commenced from the commercially available nitrobenzenesulfonyl chlorides 10–12, which reacted with KF – Et3N to give the corresponding sulfonyl fluorides 13–15 (82–93% yield). Catalytic hydrogenation of 13–15 led to the formation of anilines 16–18 (40–95% yield), which were subjected to the diazotation, followed by the Sandmeyer reaction with in situ generated SO2 to give the target compounds 1–3 in 60– 91% yield.

SO2Cl

O2N 10, o11, m12, p-

SO2F

SO2F 1. NaNO2, HCl

KF

H2

Et3N

Pd-C

O2N 13, o-, 82% 14, m-, 89% 15, p-, 93%

SO2F

2. SOCl2, H2O, CuCl ClO2S H2N 16, o-, 40% 1, o-, 60% 17, m-, 79% 2, m-, 85% 18, p-, 95% 3, p-, 91%

Scheme 2. Synthesis of the building blocks 1–3

The next part of our work included design and synthesis of the library 8{1–3,9} based on the building blocks 1–3. Initially, validation of the common sulfonamide synthesis protocol was performed for the sulfonyl halides 1–3. A sulfonamide set of 150 compounds was enumerated from 1–3 and nucleophiles 9{1–120}, followed by random selection (with at least one product per reagent included). It was found that reaction of


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1–3 with most primary and secondary (hetero)aromatic amines 9{1–102} (R1 = (het)aryl; R2 = H, alkyl, Figure 3) in the presence of pyridine as a base and CH2Cl2 as the solvent occurred slowly but more or less cleanly at rt; typically, the conversion was completed after 48 h. The products 8{1–3,1–102} were obtained with 61% synthesis success rate and 45% average yield (Table 1). Increasing the temperature to 40 C led to the loss of chemoselectivity, so that mixtures containing mono- and bis-sulfonamides, as well as unreacted starting material were formed.


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N

N N H

N

H2N 9{1}

N

N NH2

CN 9{2}

9{3}

N 9{9}

NH2

9{17}

H2N

O

HN

S

NH2

S

9{26}

O

9{35}

HCl H2N

H2N

N

9{43}

9{44}

S

N

O

9{58}

HN

N

9{65}

CN 9{66}

N

O

H N

O

HCl

N N N

HCl

NH2

NH2

9{40}

9{39}

O

9{67}

N 9{45}

9{68}

NH2

9{48}

O O

H2N

H2N

9{54}

S

O

N

S 9{55}

9{56}

O

O

H N

S

NH2

N H

NH2

9{62}

9{63} O

NH2 HN N

CN

9{64}

NH

N

N

HN N CF3 9{69}

N

O

9{61}

Cl

N N

9{47}

O S

NH2

NH2

N N

NH2

NH2

NH2

9{46}

F

9{60}

CF3

N N

O

F

N

H2N

N N

NH2 N

H2N

9{59}

O

N

HCl N

N H 9{32}

9{31}

9{38}

NH2

NH2

NH2

NH2

O

NH2

9{53}

O

O

9{57}

9{52}

N HN

H2N

N

H2N

9{51}

N

O

N N N

N

N N

S

O N

N

OPh

HN

NH

NH

N

9{24}

H2N

NH2

9{37}

H2N

N

NH

9{50}

N

9{30}

O

9{42}

NH

N N

H2N

NH

9{41}

9{29}

9{36}

HO

O

N N

HCl

O

N H

9{49}

S H2N

N N 9{23}

N

N

2HCl

H2N

O N

N

H2N

HCl

H2N 9{34}

NH2

9{16}

O H2N

9{22}

N N

O

O

9{28}

HCl

HCl

S

HCl

Cl

O 9{15}

N

9{21}

NH2

9{27}

H2N

9{33}

H2N

NH2

N

H N N

N

N

9{8}

O

9{14}

N

N N

N

N

HCl

H2N

NH2

HCl N

N H

H2N

9{20}

H2N

NH2

N

N

NH2

9{7}

9{13}

NH2

O N

9{6}

H2N

N 9{12}

9{19}

9{25}

N O

N

N H2N

O

O

N

9{18}

H N

N

9{5}

9{11}

O

N

N

H N

NH2

N

H N

O

H2N 9{4}

9{10}

NH2

N

H2N

CN N H

N

H2N

F 9{70}

NH

H2N

H2N

O 9{71}

9{72}


9


NH2

NH2

NH2

N Cl

NH2

N

Cl

NH2

9{74}

N 9{76}

F3C

9{77}

NH2

NH2

N N

H2N

Br

9{82}

N

N

9{83}

NH2

9{84} H N

NH2

N

N

N

9{90}

9{91}

N

9{85}

9{86}

9{87}

Br

NH2 9{92}

NH2

NH2

S

N

N N

9{98}

9{99}

NH2

N H2N

N

O

NH2

9{100}

HCl

N

N H

NC

NH2

NH2

NC

N

9{101}

9{102}

CN

9{107} O

9{108}

9{115}

N

9{116}

H2N

O NH2

9{131}

9{110}

N 9{104}

NH2 9{105}

NH2 HN O

H2N NH O

9{112}

9{113}

O N 9{132}

CN

9{118}

9{119}

H2N

9{125}

9{133}

N 9{126}

H N

NH2 O

NH2

O

N

CF3

NH2

H2N

CN

HCl

H2N

O

NH2

9{111}

H N

HCl H N CN

9{117}

HN N 9{124}

N H

9{109}

N H

NH2

S

2HCl N

HO

H2N

HCl

NC

HO

9{123}

F

HCl NH2

N H

NH2

9{97}

NH2 NH

O

F

9{106}

N N

HCl

F

NH2

NH2 N

NH2

9{96}

HCl

NH2

N

F

NH2

HCl

N

9{103}

F

9{89}

Cl

N

N O

H2N

9{88}

9{95}

NH2

N

HN N

N

N H

9{94}

HCl

HCl H2N

HCl

H2N

N S

NH2

NH2

N

NH2

N

9{93}

9{81}

NH2 N

N

Cl S

NH2

N

N NH

9{80}

F

H2N N

N

N

9{79}

Br

N N

HO

N

9{78}

NH2

NH2

NH2 F3C

N 9{75}

NH2

NH2 Br

F

9{73}

C l

NH2

Page 10 of 43

N N

O N

9{134}

9{135}

Br

O

N N

S N

9{121}

NH2

H2N

CN

O NH2

9{130}

9{129} H2N

F

N

9{136}

N H

9{122}

N N

N

H2N

9{128}

NH2

N

N

O

NH2

N

N

O

S 9{127}

N

CN

9{120}

H N

9{114}

O 9{137}

9{138}

Figure 3. Structures of nucleophilic reagents 9{1–138}


10

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Table 1. Parallel synthesis of sulfonyl fluorides 8

#

Product

Yield

#

Product

(%) 1

8{1,8}

0

Yield

#

Product

(%) 4

8{2,46}

99

Yield

8{1,19}

0

4

91

8{3,23}

8

8{1,24}

2

4

8{2,48}

32

92

8{3,25}

0

8{1,40}

0

4

8{2,62}

43

93

8{3,26}

0

8{1,45}

0

5

8{2,66}

61

94

8{3,27}

0

8{1,46}

24

5

8{2,70}

25

95

8{3,28}

28

8{1,50}

0

5

8{2,71}

99

96

8{3,29}

33

8{1,51}

11

5

8{2,72}

0

97

8{3,31}

43

8{1,51}

11

5

8{2,80}

0

98

8{3,32}

30

8{1,52}

0

0 1

8{2,82}

6

8{1,53}

0

2

5

8{2,85}

99

99

8{3,33}

99

0

5 7

10

8{3,34}

17

0 8{2,87}

0

6 8{1,56}

13

8{3,86}

0

13

8{3,86}

9

14

8{3,87}

75

14

8{3,88}

10

14

8{3,89}

0

14

8{3,90}

10

14

8{3,91}

9

8{3,92}

0

8{3,95}

0

8{3,96}

26

4

5

1 1

5

99

3

4 1

8{3,85}

2

3 9

13

1

2 8

99

0

1 7

8{3,85}

9

0 6

13

8

9 5

(%)

7

8 4

Yield

6

7 3

Product

(%)

6 2

#

10

5 8{3,35}

0

1 8{2,91}

7

10

14

14 6

8{3,36}

2

56

14 7


11


1

8{1,57}

0

3 1

8{1,73}

0

8{1,76}

0

8{1,77}

0

8{1,85}

0

8{1,98}

0

8{1,99}

0

8{1,102}

0

8{1,116}

0

8{1,118}

0

8{1,122}

13

8{1,123}

60

6

6

6

6

6

6

6

6

8{1,133}

74

7

8{2,108}

43

10

7

8{3,39}

19

10

10

10

8{2,109}

16

10

8{3,41}

0

43

10

8{3,44}

0

99

11

8{3,45}

99

54

11

8{3,46}

99

44

11

8{3,47}

99

37

11

8{3,48}

26

0

11

8{3,49}

0

5

11

8{3,54}

0

0

11

15

8{3,99}

21

15

8{3,100}

33

15

8{3,101}

0

15

8{3,104}

24

15

8{3,105}

0

15

8{3,107}

32

15

8{3,108}

36

8{3,108}

38

8{3,109}

30

8{3,110}

20

8{3,111}

38

7 8{3,55}

0

15 8

8{3,56}

35

15 9

8{3,58}

0

5 8{3,1}

99

6

4 8{2,134}

8{3,98}

5

3 8{2,119}

15

4

2 8{2,115}

99

3

1 8{2,114}

8{3,98}

2

0 8{2,113}

14

1

9 8{2,112}

99

0

8 8{2,110}

8{3,97}

9

7

0 8{1,137}

10

14 8

6

9

5 2

23

8

4 2

8{2,106}

7

3 2

6

23

5

6

2 2

99

5

1 2

8{2,97}

4

0 2

6

8{3,37}

4

3

9 2

0

2

8 1

8{2,94}

1

7 1

5

10 3

0

6 1

23

9

5 1

8{2,93}

8

4 1

5

Page 12 of 43

16 0

8{3,59}

0

16


12

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6 2

1 8{1,138}

37

7 2

8{1,140}

38

8{2,3}

59

8{2,4}

58

8{2,8}

0

8{2,15}

35

8{2,18}

48

8{2,19}

56

8{2,22}

12

8{2,23}

8

8{2,28}

15

9

26

7

7

7

7

8

8

8

8{2,29}

27

8

8{3,6}

0

57

8 4

8{3,61}

0

11

12

12

8{3,7}

14

12

8{3,63}

0

46

12

8{3,64}

0

0

12

8{3,65}

0

12

12

8{3,67}

0

0

12

8{3,68}

99

99

12

8{3,69}

28

99

12

8{3,74}

46

23

12

0

16

8{3,119}

0

16

8{3,120}

0

16

8{3,121}

75

16

8{3,124}

99

16

8{3,125}

99

17

8{3,126}

8

8{3,127}

42

8{3,128}

72

8{3,129}

99

8{3,130}

61

0 8{3,75}

52

17 1

8{3,76}

55

17 2

8{3,76}

55

8 8{3,15}

8{3,117}

9

7 8{3,14}

16

8

6 8{3,13}

31

7

5 8{3,12}

8{3,114}

6

4 8{3,10}

16

5

3 8{3,9}

53

4

2 8{3,8}

8{3,112}

3

1

3 8{2,30}

11

16 2

0

2

8 3

8{3,5}

1

7 3

7

43

9

0

6 3

68

9

5 3

8{3,4}

8

4 3

7

8{3,60}

8

7

3 3

66

6

2 3

8{3,3}

5

1 3

7

11

1

7

4

0 3

0

3

9 3

8{3,2}

2

8 2

7

6

17 3

8{3,77}

9

99

17 4


13


4

8{2,32}

30

0 4

8{2,34}

35

8{2,36}

59

8{2,38}

97

5

8{3,17}

23

8

8

8{2,42}

51

8

8{3,18}

30

99

9 0

3

13

13

8{3,19}

22

13

8{3,79}

38

0

13

8{3,81}

34

30

13

23

17

8{3,132}

7

17

8{3,135}

10

8{3,136}

74

8{3,137}

46

7 8{3,82}

0

17 8

8{3,83}

0

4 8{3,21}

8{3,131}

6

3 8{3,20}

17 5

2

9 8{2,43}

8{3,78}

1

8

4 4

8

13 0

7

3 4

0

6

2 4

8{3,16}

5

1 4

8

Page 14 of 43

17 9

8{3,84}

27

5

Low chemoselectivity was also observed with primary amines 9{103–106}, including sterically hindered tert-butylamine 9{103}, even if the reaction was performed at room temperature (although we managed to isolate the library members 8{2,104} and 8{2,106}, both in 23% yield, after chromatographic purification). Nevertheless, the method could be extended to less nucleophilic aliphatic amines bearing electronwithdrawing groups (e.g. -alkyl -amino nitriles 9{107–109} or fluorinated amines 9{110,111}, as well as acyl hydrazides 9{112–114} and phenols 9{115}, although in these cases, isolated yields were slightly lower (average value is 38%). The procedure


14

Page 15 of 43 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


did not work with the parent N-(cyanomethyl)amines 9{116–118} and ,-dialkyl amino nitriles 9{119,120}: mixtures of unidentified products were obtained in these experiments.

Therefore, the developed procedure for the synthesis of SO2F-containing sulfonamides 8{1–3,9} can be effective for (hetero)aromatic, as well as aliphatic amines with low nucleophilicity, hydrazides, and phenols. The method tolerates azole, azinone, and carboxamide N–H bonds, hydroxyl, nitrile, and aromatic bromide functions, as well as 2- or 4-chloropyridine moieties. 92 of 150 library members 8{1–3,9} were prepared in the parallel synthesis during the validation step (61% synthesis success rate, 43% average yield), although in most cases, chromatographic purification of the products was necessary. Analysis of unsuccessful results obtained with aromatic amines did not reveal any additional regularities; in our opinion, the reasons behind these failures might be related to isolation and/or purification issues and not chemical problems.

The following order of efficiency in the synthesis of the library 8 was observed for the sulfonyl halides 1–3 according to the synthesis success rate: 2 (87%) > 3 (61%) > 1


15


Page 16 of 43

(14%). Poor results obtained with o-isomer 1 might be attributed to the limited stability of the final products 8{1,9} towards hydrolysis (possibly due to the participation of the osulfonamide moiety) since in most cases, the corresponding sulfonic acid was observed as the main component in the crude products according to LCMS data. These conclusions are supported by the results obtained from stability of the compounds 8 in DMSO solutions: whereas in the case of the products obtained from the sulfonyl halides 2 and 3, the compounds were stable at rt for several days, the compounds 8{1,9} showed considerable degradation at these conditions after overnight storage. Therefore, it is preferable to use their freshly prepared solutions.

Further extension of the library 8 was aimed at demonstrating its potential to inhibit serine proteases using the classical target of this class – trypsin – as a model enzyme. The library enumeration followed the principles of the REAL concept published elsewhere.22 In particular, virtual coupling of sulfonyl halides 1–3 and 1224 (hetero)aromatic or fluorinated aliphatic amines, or -aminonitriles 9 with in-house validated reactivity in the sulfonamide bond formation, afforded 31224 = 3672 synthesizable library members. These were subjected to covalent docking into the


16

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trypsin S1 pocket that is quite deep and planar so that comfortable accommodation of aromatic cores is possible.23 The catalytically active Ser195 was responsible for the covalent bond formation; in addition to that, several hydrogen bond donors/acceptors were also present in the binding site, including side chains of Asp189, Ser190, Gln192, as well as backbone NH groups of Ser195, Gly193 and Gly216.

As a result of the docking procedure, 100 compounds were selected as virtual hits according to the following criteria: the built-in QXP scoring function,24 the number of hydrogen bonds (was exposed to a minimum of four hydrogen bonds), the proteinligand contact surface area and the distances from ligand to the key points of the corresponding pharmacophore model (Figure 4). The model was based on the limited ligand mobility due to the covalent bond with Ser195 and the structural features of the entire series of compounds. Formation of hydrogen bonds between oxygen atoms of the two sulfo groups and Gly193, as well as Gln192 and/or Gly216, respectively, was desirable. An additional hydrogen bond with Gly219 might be also present for the compounds bearing a sulfonamide NH group. This interaction fixed the (het)aryl fragment L2 in the S1 sub-pocket; filling the sub-pocket and approaching a part of the


17


Page 18 of 43

ligand to Asp 189 was considered as one of the selection criteria. A separate filter removed ligands strongly exposed to the solution. Further processing of the complexes was carried out by visual inspection according to the assumed pharmacophore interaction model.

Figure 4. Pharmacophore model of ligand binding in the S1 pocket of trypsin (red and blue spheres show hydrogen bond donors and acceptors, respectively; L1, L2 denote aromatic/hydrophobic moieties

62 of 100 sulfonyl fluorides from the resulting set were synthesized using the developed procedure (Table S1). These products were subjected to in vitro screening against trypsin, including protein thermal shift assay (TSA),25–27 followed by BApNA (N-


18

Page 19 of 43 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


benzoyl-DL-arginine p-nitroanilide) enzymatic digestion assay28 (Table 2). It is interesting to note that most of the identified hits demonstrated negative thermal shift. This indicates destabilization of the folded protein in the presence of the ligand,29 and is sometimes perceived as an undesirable feature of the TSA hits.30,31 Nevertheless, three of these compounds had IC50 values lower than the standard (PMSF) in the enzymatic digestion assay, the compound 8{1,122} being five-fold more active than PMSF (Figure 5); other showed activity comparable to that of PMSF. It should be noted that all the identified hits were rather polar (cLogP = 0.77–2.6032), which might be useful to satisfy the hydrogen-bonding potential of the compounds in the protein binding site. In particular, the ligand-lipophilicity efficiency (defined as LLE = pIC50 – cLogP33) for the most active compound 8{1,122} (3.28) was improved as compared to that of PMSF (1.93). These data demonstrate that the library 8{1–3,9} can be a promising tool for discovery of covalent modifiers.

Table 2. Results of in vitro testing of the sulfonyl fluorides 8 against trypsin

Entry No.

Compound

Tm, C IC50 (M)

cLogP32

LLE


19


O F S O

Page 20 of 43

–9.5

338

2.23

1.24

–10.6

224

2.60

1.05

–7.8

648

2.09

1.10

0.16

795

1.37

1.73

5

O N S O O S F O Z1737943418 8{1,51}

–3.0

1721

2.34

0.42

6

O N S S O N N O S F O Z2216296226 8{1,122}

–3.6

84

0.80

3.28

7

O N S N O N O S F O Z2216296225 8{1,123}

–12.0

988

0.77

2.24

1

N

O S N H O Z1672278953 8{3,74}

O

2

N

O S N H O

S

O

O F S O

Z1672278926 8{3,56}

3

N S

O F S O

O S N H O Z1672278918 8{3,98}

O

4

F S

O O S N O

N N

Z2216296236 8{1,140}


20

Page 21 of 43 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


8

O S F O

–0.11

480

1.39

1.93

PMSF

Figure 5. Dose-response curves for the most active compounds 8{1,122} and 8{3,56} compared to those for PMSF (red curves are the same)

Analysis of the binding mode for the two most active compounds 8{1,122} and 8{3,56} showed that in the case of 8{3,56}, the ligand fit well with the pharmacophore model shown in Figure 4. In particular, the sulfo groups forms hydrogen bonds with Gln192, Gly216, and Gly193; and the hydrogen bond was also present between the sulfonamide NH and Gly219 (Figure 6). A different binding mode was observed for 8{1,122}: while the key hydrogen bonds with Gln192 and Gly193 were retained, the sulfo group attached to Ser195 also formed hydrogen bond with catalytic His57. In


21


Page 22 of 43

addition to that, the N-4 atom of the thiadiazole ring formed a hydrogen bond with Ser214 buried in the S1 sub-pocket. In both cases, the corresponding heteroaromatic moieties filled the S1 sub-pocket of trypsin, as it was suggested by the pharmacophore model.

8{1,122}

8{3,56}

Figure 6. Binding poses of compounds 8{1,122} and 8{3,56} in the S1 pocket of trypsin according to the docking results

Conclusions.

The difference in reactivity of sulfonyl halides can be used to selectively modify the sulfonyl chloride moiety in the presence of the sulfonyl fluoride one within the same molecule.

We

demonstrated

this

feature

for

three

isomeric


22

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(chlorosulfonyl)benzenesulfonyl fluorides. Therefore, these bifunctional building blocks are convenient starting points to generate the libraries of sulfonyl fluorides. The scope of the developed parallel synthesis procedure includes various (hetero)aromatic, as well as electron-poor aliphatic amines (i.e. -amino nitriles or fluorinated amines). The library of the obtained sulfonyl fluorides is a promising tool for design of covalent modifiers targeting serine/threonine and lysine residues in the protein molecules, which is validated by discovery of several trypsin inhibitors having two- and five-times higher activity compared with the known protease inhibitor PMSF.

Experimental part.

General. All chemicals and solvents were obtained from Enamine Ltd. and used without further purification. 1H and

13C

NMR spectra were acquired on Bruker Advance

DRX 400 and Bruker Avance DRX 500 spectrometers using DMSO-d6 as a solvent and tetramethylsilane as an internal standard. Melting points were determined on a Buchi melting point apparatus. LC-MS data were recorded on Agilent 1100 HPLC equipped with diode-matrix and mass-selective detector Agilent LC/MSD SL, column: Zorbax SB-


23


Page 24 of 43

C18, 4.6 mm × 15 mm; eluent: A, acetonitrile – H2O with 0.1% of TFA (95:5), B, H2O with 0.1% of TFA; flow rate: 1.8 mL/min. Elemental analyses were performed at the Laboratory of Organic Analysis, Department of Chemistry, Kyiv National Taras Shevchenko University. Preparative flash chromatography was performed on a Combiflash Companion chromarograph (12 g RediSep columns, gradient Hexanes – i-PrOH as eluent). Synthesis of sulfonyl fluorides 13–15. To a solution of sulfonyl chloride 10–12 (103 g, 0.467 mol) in CH2Cl2 (1000 mL), solutions of KF (81.3 g, 1.40 mol) in H2O (400 mL) and Et3N (78.1 mL, 0.560 mol) in CH2Cl2 (400 mL) were added. The mixture was stirred at room temperature for 4 h, then the organic phase was separated, washed with H2O (400 mL), saturated aq NaHCO3 (400 mL) and evaporated in vacuo. 2-Nitrobenzenesulfonyl fluoride (13). Yield 78.6 g, 82%. For spectral and physical data, see ref.8,34 3-Nitrobenzenesulfonyl fluoride (14). Yield 85.3 g, 89%. For spectral and physical data, see ref.35 4-Nitrobenzenesulfonyl fluoride (15). Yield 89.1 g, 93%. For spectral and physical data, see ref.34,35 Synthesis of sulfonyl fluorides 16–18. Nitrobenzene derivative 16–18 (78.2 g, 0.381 mol) was dissolved in MeOH (750 mL), and 5% Pd-C (12.5 g) was added. The resulting mixture was hydrogenated in an autoclave at 50 C for 72 h. The catalyst was filtered off, and the combined filtrates were evaporated in vacuo. The residue was triturated with t-BuOMe (750 mL). The


24

Page 25 of 43 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


solution was decanted from the residue and dried over Na2SO4. The solvent was removed under reduced pressure, and the residue was dried in vacuo. 2-Aminobenzenesulfonyl fluoride (16). Yield 26.7 g, 40%. For spectral and physical data, see ref.36 3-Aminobenzenesulfonyl fluoride (17). Yield 52.7 g, 79%. Colorless liquid. 1H NMR (400 MHz, CDCl3) δ 7.45 – 7.31 (m, 2H), 7.22 (s, 1H), 6.98 (d, J = 6.9 Hz, 1H), 4.03 (s, 2H).

13C

NMR (125 MHz, CDCl3) δ 147.6, 133.7 (d, J = 23.2 Hz), 130.5, 121.4, 117.6, 113.3. 19F NMR (376 MHz, CDCl3) δ 64.6. LC/MS (EI): m/z = 176 [M+H]+. Anal. calcd. for C6H6FNO2S: C, 41.14; H, 3.45; N, 8.00; S, 18.30. Found: C, 41.03; H, 3.49; N, 7.97; S, 17.95. 4-Aminobenzenesulfonyl fluoride (18). Yield 63.4 g, 95%. For spectral and physical data, see ref.37 Synthesis of sulfonyl fluorides 1–3. Thionyl chloride (41.0 mL, 0.564 mol) was added dropwise to ice (250 g). The mixture was left for 48 h, then cooled to –5 C, and CuCl (0.574 g, 5.80 mmol) was added to give the solution A. Sulfonyl fluoride 16–18 (25.6 g, 0.146 mol) was dissolved in 20% aq HCl (250 mL), the solution was cooled to –10 C, and a solution of NaNO2 (12.1 g, 0.175 mol) in H2O (29.0 mL) was added dropwise. The resulting solution was stirred at room temperature for 20 min, cooled to –5 C, and added dropwise to a solution A. The mixture was stirred at –5 C for 3 h and then filtered. The filtrates were extracted with CH2Cl2 (3250 mL), the combined extracts were dried over Na2SO4 and evaporated in vacuo. The residue was recrystallized from hexanes. 2-(Chlorosulfonyl)benzenesulfonyl fluoride (1). Yield 22.6 g, 60%. Orange crystals; mp = 88–90 C. 1H NMR (400 MHz, CDCl3) δ 8.46 (d, J = 7.4 Hz, 1H), 8.41 (d, J = 7.7 Hz, 1H), 8.03 (quint, J = 2.7 Hz, 2H).

13C

NMR (125 MHz, CDCl3) δ 142.5, 136.3, 135.7, 133.3 (d, J = 1.9

Hz), 132.3, 131.3 (d, J = 28.6 Hz). 19F NMR (376 MHz, CDCl3) δ 66.0. GC/MS (EI): m/z = 258


25


Page 26 of 43

[M]+. Anal. calcd. for C6H4ClFO4S2: C, 27.86; H, 1.56; S, 24.79; Cl, 13.71. Found: C, 28.24; H, 1.65; S, 24.67; Cl, 14.02. 3-(Chlorosulfonyl)benzenesulfonyl fluoride (2). Yield 32.1 g, 85%. Yellowish crystals; mp = 27–29 C. 1H NMR (400 MHz, CDCl3) δ 8.64 (s, 1H), 8.43 (d, J = 8.6 Hz, 1H), 8.36 (d, J = 8.1 Hz, 1H), 7.96 (t, J = 8.1 Hz, 1H). 13C NMR (125 MHz, CDCl3) δ 145.8, 135.2 (d, J = 28.1 Hz), 134.3, 133.4, 131.7, 127.1.

19F

NMR (470 MHz, DMSO-d6) δ 66.2. LC/MS (EI): m/z = 258

[M]+. Anal. calcd. for C6H4ClFO4S2: C, 27.86; H, 1.56; S, 24.79; Cl, 13.71. Found: C, 27.96; H, 1.78; S, 24.82; Cl, 13.47. 4-(Chlorosulfonyl)benzenesulfonyl fluoride (3). Yield 34.4 g, 91%. Yellowish powder; mp = 109–111 C. 1H NMR (400 MHz, CDCl3) δ 8.33 (d, J = 8.4 Hz, 2H), 8.29 (d, J = 8.4 Hz, 2H). 13C

NMR (100 MHz, CDCl3) δ 149.6, 138.9 (d, J = 27.4 Hz), 130.0, 128.3. 19F NMR (376 MHz,

CDCl3) δ 65.6. LC/MS (EI): m/z = 258 [M]+. Anal. calcd. for C6H4ClFO4S2: C, 27.86; H, 1.56; S, 24.79; Cl, 13.71. Found: C, 27.95; H, 1.75; S, 24.48; Cl, 13.69. Parallel synthesis of sulfonyl fluorides 8. Amine 9 (1 mmol), pyridine (0.5 mL), CHCl3 (1 mL), and sulfonyl chloride 1–3 (258 mg, 1 mmol) were placed into a vial, and the mixture was shaken at room temperature for 48 h. Then it was evaporated in vacuo, CHCl3 (3 mL) was added, the solution was washed with H2O (1 mL), and evaporated in vacuo. The residue was purified by preparative flash chromatography. 3-(N-(Pyridin-4-yl)sulfamoyl)benzene-1-sulfonyl fluoride (9{2,85}). Yield 86 mg, quant. Yellowish solid; mp = 207–209 C. 1H NMR (400 MHz, DMSO-d6) δ 12.98 (br s, 1H), 8.41 – 8.20 (m, 3H), 8.11 – 8.02 (m, 2H), 7.93 (t, J = 7.8 Hz, 1H), 7.02 (d, J = 7.2 Hz, 2H). 13C NMR (125 MHz, DMSO-d6) δ 162.5, 146.5, 139.8, 134.1, 132.6 (d, J = 24.2 Hz), 132.2, 131.4, 125.6, 115.5. 19F NMR (376 MHz, DMSO-d6) δ 66.1. LC/MS (CI): m/z = 317 [M+H]+. Anal. calcd. for C11H9FN2O4S2: C, 41.77; H, 2.87; N, 8.86; S, 20.27. Found: C, 41.69; H, 2.92; N, 9.14; S, 20.56.


26

Page 27 of 43 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


Virtual screening. Molecular docking was performed using a flexible ligand and a fixed receptor model. We used an algorithm of MCDOCK (Monte Carlo search) implemented in QXP docking software, which had shown high reproducing ability of ligand conformation with minimum RMSD in comparison to the crystallographic data, but more time-consuming than standard methods.38 The binding site model was created on the basis of PDB 2PLX.23 When converting the structure of the protein and creating a binding site model, the center was exposed on Ser 190. The sulfonyl fluorides database was prepared for the dock combinatorial library. The maximum number of MCDOCK routine steps was set to 500, and the 10 best structures (based on built-in QXP scoring function)24 were retained for each compound. The resulting protein-ligand complex structures had been filtered by intrinsic Flo+ filters and multiRMSD software package.39

In vitro screening. Reference compounds – PMSF (phenylmethyl sulfonyl fluoride) and benzamidine – were obtained from Enamine Ltd. (Kyiv, Ukraine). Stock solutions of the tested compounds were prepared at 20 mM in DMSO and were stored at –20 °C until use. All thermal shift assay (TSA) experiments with trypsin (Sigma, Cat. T8003) were performed using ViiA™7 real-time PCR System equipped with 384-well heat block


27


Page 28 of 43

(Applied Biosystems, USA). General TSA methodology was adopted from the literature24-26,31 and experimentally modified in order to optimize conditions for measuring trypsin melting temperature shifts upon interaction with small molecules. To define the optimal buffer composition for the TSA procedure, a matrix of common biological buffers combined on a 96-well microplate, including phosphate, acetate, TRIS, HEPES, MES at different pH and inorganic salts content, was tested in thermal melt experiment on a 384-well microplate (each buffer composition in quadruplicate). In addition, each buffer composition was tested at two different buffering component concentrations, resulting in two 384-well plate buffer screening experiments and the total of 192 different buffer compositions tested. Selection of the optimal buffer composition was based on balancing two criteria including maximized melting temperature (thermal stability) of trypsin and maximized melting temperature shift induced by the benzamidine at the concentration of 500 µM. As a result, the buffer consisting of 20 mM Tris-HCl pH 7.0 and 1 mM CaCl2 was selected for the screening. Trypsin was pre-mixed with SYPRO Orange dye (Thermo Fischer Scientific, Cat. S6650, 5000 stock) to prepare a master mix at 345 µg/mL


28

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protein and 10 dye concentration. PMSF and Benzamidine were used as reference compounds at the final concentrations of 200 uM and 500 uM, respectively. Tested compounds were added to the protein-dye master mix at 200 uM (1% DMSO concentration) and incubated at room temperature for 15 min in MicroAmp® optical 384well reaction plates (ThermoFisher, Cat. 4309849) sealed with optical sealing film (ThermalSeal RT2, Excel Scientific, Cat. TS-RT2). The volumes of all reaction mixtures were 10 µL (3.45 µg of trypsin per well). Thermal scanning was performed by raising temperature to 34 °C at 2.5 °C/min without signal detection followed by 34 °C to 75 °C temperature ramp at 0.1 °C/s with constant fluorescence intensity reading at 1 sec intervals using EX470/EM623 nm filter set. Screening of compounds was carried out in quadruplicate (n = 4). The raw data of dye fluorescence intensity change upon protein melt

were

exported

using

the

ViiA7

RUO

software

1.2

(Applied

Biosystems/ThermoFischer Scientific). Further data visualization, curve fitting, melting temperature calculations on the raw fluorescence data were performed using custommade Microsoft Excel scripts. The peak of the first derivative for the fluorescence curve was used to define melting temperature (Tm). Averaged Tm values for the control wells


29


Page 30 of 43

(n = 64 per plate), were used as a reference point to determine melting temperature shifts of compounds (ΔTm). Enzymatic trypsin assay using chromogenic BApNA substrate (Sigma-Aldrich, Cat. B4875) was

performed

using

the

procedures

reported

previously28

and

recommended by the manufacturer.

Supporting Information Available: Table S1, and copies of NMR spectra. This material is available free of charge via the Internet at http://pubs.acs.org.

Acknowledgments. The authors thank Prof. Andrey A. Tolmachev for his encouragement and support, Mr. Bohdan Vashchenko and Ms. Yuliya Kuchkovska for their help with manuscript preparation.

References and footnotes.

(1)

Dong, J.; Krasnova, L.; Finn, M. G.; Sharpless, K. B. Sulfur(VI) Fluoride Exchange (SuFEx): Another Good Reaction for Click Chemistry. Angew. Chem. Int. Ed. 2014, 53 (36), 9430–9448.


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(2)

Narayanan, A.; Jones, L. H. Sulfonyl Fluorides as Privileged Warheads in Chemical Biology. Chem. Sci. 2015, 6 (5), 2650–2659.

(3)

Powers, J. C.; Asgian, J. L.; Ekici, Ö. D.; James, K. E. Irreversible Inhibitors of Serine, Cysteine, and Threonine Proteases. Chem. Rev. 2002, 102 (12), 4639– 4750.

(4)

Kathman, S. G.; Statsyuk, A. V. Covalent Tethering of Fragments for Covalent Probe Discovery. Med. Chem. Commun. 2016, 7 (4), 576–585.

(5)

Zhersh, S. A.; Blahun, O. P.; Sadkova, I. V.; Tolmachev, A. A.; Moroz, Y. S.; Mykhailiuk, P. K. Saturated Heterocyclic Aminosulfonyl Fluorides: New Scaffolds for Protecting-Group-Free Synthesis of Sulfonamides. Chem. Eur. J. 2018, 24 (33), 8343–8349.

(6)

Semenok, D.; Kletskov, A.; Dikusar, E.; Potkin, V.; Lukin, O. Efficient Synthesis of Chalcone-4′-Sulfonyl Chlorides and Fluorides. Tetrahedron Lett. 2018, 59 (4), 372–374.

(7)

Han, L.-C.; Stanley, P. A.; Wood, P. J.; Sharma, P.; Kuruppu, A. I.; Bradshaw, T. D.; Moses, J. E. Horner–Wadsworth–Emmons Approach to Piperlongumine


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Analogues with Potent Anti-Cancer Activity. Org. Biomol. Chem. 2016, 14 (31), 7585–7593. (8)

Mukherjee, H.; Debreczeni, J.; Breed, J.; Tentarelli, S.; Aquila, B.; Dowling, J. E.; Whitty, A.; Grimster, N. P. A Study of the Reactivity of S

(VI)

–F Containing

Warheads with Nucleophilic Amino-Acid Side Chains under Physiological Conditions. Org. Biomol. Chem. 2017, 15 (45), 9685–9695. (9)

Fadeyi, O.; Parikh, M. D.; Chen, M. Z.; Kyne, R. E.; Taylor, A. P.; O’Doherty, I.; Kaiser, S. E.; Barbas, S.; Niessen, S.; Shi, M.; Weinrich, S. L.; Kath, J. C.; Jones, L. H.; Robinson, R. P. Chemoselective Preparation of Clickable Aryl Sulfonyl Fluoride Monomers: A Toolbox of Highly Functionalized Intermediates for Chemical Biology Probe Synthesis. ChemBioChem 2016, 17 (20), 1925–1930.

(10) Wang, P.; Dong, Y.; Lu, X.; Wu, Z.; Chen, H. Combining Click Sulfur(VI)-Fluoride Exchange with Photoiniferters: A Facile, Fast, and Efficient Strategy for Postpolymerization Modification. Macromol. Rapid Commun. 2018, 39 (3), 1700523. (11) Zhao, Q.; Ouyang, X.; Wan, X.; Gajiwala, K. S.; Kath, J. C.; Jones, L. H.;


32

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Burlingame, A. L.; Taunton, J. Broad-Spectrum Kinase Profiling in Live Cells with Lysine-Targeted Sulfonyl Fluoride Probes. J. Am. Chem. Soc. 2017, 139 (2), 680– 685. (12) Leng, J.; Qin, H.-L. 1-Bromoethene-1-Sulfonyl Fluoride (1-Br-ESF), a New SuFEx Clickable Reagent, and Its Application for Regioselective Construction of 5Sulfonylfluoro Isoxazoles. Chem. Commun. 2018, 54 (35), 4477–4480. (13) Bogolubsky, A. V.; Moroz, Y. S.; Mykhailiuk, P. K.; Pipko, S. E.; Konovets, A. I.; Sadkova, I. V.; Tolmachev, A. Sulfonyl Fluorides as Alternative to Sulfonyl Chlorides in Parallel Synthesis of Aliphatic Sulfonamides. ACS Comb. Sci. 2014,

16 (4), 192–197. (14) Hart, R. M.; Whitehead, M. A. 35Cl n.q.r. of Fifty-Nine Molecules Containing the S–Cl Bond. Trans. Faraday Soc. 1971, 67 (0), 3451–3464. (15) Petrow, V.; Stephenson, O.; Wild, A. M. Studies In The Field Of Diuretic Agents.

J. Pharm. Pharmacol. 1960, 12 (1), 705–719. (16) Steinkopf, W. Über Aromatische Sulfofluoride. J. Prakt. Chem. 1927, 117 (1), 1– 82.


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Page 34 of 43

(17) Baker, B. R.; Janson, E. E. Irreversible Enzyme Inhibitors. CLV. Active-SiteDirected Irreversible Inhibitors of Dihydrofolic Reductase Derived from 1-[4.Omega.-Aminoalkoxy)-3-Chlorophenyl]-4,6-Diamino-1,2-Dihydro-2,2-Dimethyl-sTriazines Bearing a Terminal Sulfonyl Fluoride. J. Med. Chem. 1969, 12 (4), 672– 676. (18) Corriu, R. J. P.; Lanneau, G. F.; Perrot-Petta, M.; Nehta, V. D. A New and Mild Access to N-Functionalized Formamido and Thioformamido Compounds Using Hypervalent Silicon Hydrides. Tetrahedron Lett. 1990, 31 (18), 2585–2588. (19) Lee, C.; Field, L. Sulfinic Acids and Related Compounds 23. Preparation of Sulfinic Acids by the Reaction of Sulfonyl Halides with Thiols. Synthesis 1990,

1990 (05), 391–397. (20) Van der Puy, M. Potassium Fluoride Catalyzed Fluorodesulfonylations of Aryl Sulfonyl Fluorides. J. Org. Chem. 1988, 53 (18), 4398–4401. (21) Photographic Color Couplers. Belg. Pat. BE634665, 1964. Chem. Abstr. 1965, 62:3485. (22) Shivanyuk, A.; Ryabukhin, S. V.; Bogolubsky, A. V.; Tolmachev, A. Enamine Real


34

Page 35 of 43 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


Database: Making Chemical Diversity Real. Chem. Today 2007, 25 (6), 58–59. (23) Conners, R.; Konarev, A. V; Forsyth, J.; Lovegrove, A.; Marsh, J.; Joseph-Horne, T.; Shewry, P.; Brady, R. L. An Unusual Helix-Turn-Helix Protease Inhibitory Motif in a Novel Trypsin Inhibitor from Seeds of Veronica (Veronica Hederifolia L.). J.

Biol. Chem. 2007, 282 (38), 27760–27768. (24) McMartin, C.; Bohacek, R. S. QXP: Powerful, Rapid Computer Algorithms for Structure-Based Drug Design. J. Comput. Aided. Mol. Des. 1997, 11 (4), 333– 344. (25) Lo, M.-C.; Aulabaugh, A.; Jin, G.; Cowling, R.; Bard, J.; Malamas, M.; Ellestad, G. Evaluation of Fluorescence-Based Thermal Shift Assays for Hit Identification in Drug Discovery. Anal. Biochem. 2004, 332 (1), 153–159. (26) Matulis, D.; James K. Kranz; F. Raymond Salemme, A.; Todd, M. J. Thermodynamic Stability of Carbonic Anhydrase: Measurements of Binding Affinity and Stoichiometry Using ThermoFluor. Biochemistry 2005, 44, 5258– 5266. (27) Niesen, F. H.; Berglund, H.; Vedadi, M. The Use of Differential Scanning


35


Page 36 of 43

Fluorimetry to Detect Ligand Interactions That Promote Protein Stability. Nat.

Protoc. 2007, 2 (9), 2212–2221. (28) Erlanger, B. F.; Kokowsky, N.; Cohen, W. The Preparation and Properties of Two New Chromogenic Substrates of Trypsin. Arch. Biochem. Biophys. 1961, 95 (2), 271–278. (29) Cimmperman, P.; Baranauskienė, L.; Jachimovičiūtė, S.; Jachno, J.; Torresan, J.; Michailovienė, V.; Matulienė, J.; Sereikaitė, J.; Bumelis, V.; Matulis, D. A Quantitative Model of Thermal Stabilization and Destabilization of Proteins by Ligands. Biophys. J. 2008, 95 (7), 3222–3231. (30) Scott, D. E.; Ehebauer, M. T.; Pukala, T.; Marsh, M.; Blundell, T. L.; Venkitaraman, A. R.; Abell, C.; Hyvönen, M. Using a Fragment-Based Approach To Target Protein–Protein Interactions. ChemBioChem 2013, 14 (3), 332–342. (31) Holvey, R. S.; Valkov, E.; Neal, D.; Stewart, M.; Abell, C. Selective Targeting of the

TPX2

Site

of

Importin-α

Using

Fragment-Based

Ligand

Design.

ChemMedChem 2015, 10 (7), 1232–1239. (32) Instant JChem, Version 17.2.27.0. ChemAxon ((http://www.chemaxon.com),


36

Page 37 of 43 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60


2018. (33) Cavalluzzi, M. M.; Mangiatordi, G. F.; Nicolotti, O.; Lentini, G. Ligand Efficiency Metrics in Drug Discovery: The Pros and Cons from a Practical Perspective.

Expert Opin. Drug Discov. 2017, 12 (11), 1087–1104. (34) Tang, L.; Yang, Y.; Wen, L.; Yang, X.; Wang, Z. Catalyst-Free Radical Fluorination of Sulfonyl Hydrazides in Water. Green Chem. 2016, 18 (5), 1224– 1228. (35) Hsu, Y. L.; Yang, C. C.; Chou, T. C.; Tai, C. H.; Chen, L. Y.; Fu, S. L.; Lin, J. J.; Lo, L. C. Design, Synthesis, and Evaluation of Cell Permeable Probes for Protein Kinases. Tetrahedron 2016, 72 (1), 58–68. (36) Schaefer, T.; Parr, W. J. E. Long-Range ’H, 19F Spin-Spin Coupling Constants in Benzenesulfonyl Fluoride and Some of Its Derivatives. Insensitivity of 19F Shifts to Ortho Substituents. J. Can. Chim. 1978, 56 (13), 1717–1720. (37) Barrow, A. S.; Moses, J. E. Synthesis of Sulfonyl Azides via Lewis Base Activation of Sulfonyl Fluorides and Trimethylsilyl Azide. Synlett 2016, 27 (12), 1840–1843.


37


Page 38 of 43

(38) Warren, G. L.; Andrews, C. W.; Capelli, A.-M.; Clarke, B.; LaLonde, J.; Lambert, M. H.; Lindvall, M.; Nevins, N.; Semus, S. F.; Senger, S.; Tedesco, G.; Wall, I. D.; Woolven, J. M.; Peishoff, C. E.; Head, M. S. A Critical Assessment of Docking Programs and Scoring Functions. J. Med. Chem. 2006, 49 (20), 5912–5931. (39) Sudakov, O. O.; Balinskyi, O. M.; Platonov, M. O.; Kovalskyy, D. B. Geometric Filters for Protein–ligand Complexes Based on Phenomenological Molecular Models. Biopolym. Cell 2013, 29, 395–401.


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Figure 4 279x209mm (96 x 96 DPI)



Figure 5a 50x31mm (300 x 300 DPI)


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Figure 5b 49x31mm (300 x 300 DPI)



Figure 6a 244x183mm (300 x 300 DPI)


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Figure 6b 244x183mm (300 x 300 DPI)


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