Sulfur-Fluoride Exchange (SuFEx)kanai/seminar/pdf/Lit_K_Maruyama_M2.pdfSuFEx Reaction 5 General...
Transcript of Sulfur-Fluoride Exchange (SuFEx)kanai/seminar/pdf/Lit_K_Maruyama_M2.pdfSuFEx Reaction 5 General...
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1
Literature Seminar
2019/11/28
M2 Katsuya Maruyama
Sulfur-Fluoride Exchange
(SuFEx)
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Contents
2
- Introduction
- Synthesis of S(VI)-F unit
- Applications of SuFEx
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K. B. Sharpless
3
Nobel Laureate K. Barry Sharpless became W. M. Keck Professor
of Chemistry at The Scripps Research Institute and The Skaggs
Institute of Chemical Biology in 1990. Previously a professor at
MIT and Stanford, he was educated at Dartmouth College (BA
1963), Stanford (PhD 1968 with E. E. van Tamelen; postdoc 1969
with J. P. Collman), and Harvard (postdoc 1970, K. E. Bloch).
The Nobel Prize in Chemistry (2001):
"for his work on chirally catalyzed oxidation reactions."
Asymmetric epoxidation Asymmetric dihydroxylation
Asymmetric aminohydroxylation
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Click Chemistry
4
- Robust connection
- High functional group tolerance
- Fast kinetics
- Easy operation
Nat. Rev. Chem. 2018, 2, 202.
2000 2001 2002 2007 2008 2014
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SuFEx Reaction
5
General scheme
Strong S
Fluoride radical is energetically inaccessible
two partners offer versatile ways to make and activate SO2F bonds
Stable S-F bond (necessity of activation)
Metal free
diversity of linkage
“Context dependent” Click reaction
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Property of Sulfonyl Fluoride
Sharpless, K. et al. Angew. Chem. Int. Ed. 2014, 53, 9430.
Substitution at S(VI) center
(no fluorination)
Heating
Oxidation/reduction
Nucleophilic attack
Activation with water
BDE
SO2F2 90.5 kcal/mol
SO2Cl2 46 kcal/mol
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7
SuFEx Reaction
Activation of S(VI)-F
- Hydrogen bond donor (acid, HX)
- Silylated nucleophile
Reactivity: Et3N < DBU < BEMP
より)HB of HF2
-: 40 kcal/mol
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SuFEx Reaction
Activation with Ca(NTf)2 (Lewis acid) Proposed mode of activation
Am Ende, C. W.; Ball, N. D. et al. Org. Lett. 2018, 20, 3943.
Effect of Ca salt Selectivity of nucleophile
JOC 2017, 82, 2294.
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9
Synthesis of SuFEx unit
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SuFEx Unit
10Chinthakindi, P. K.; Arvidsson, P. I. Eur. J. Org. Chem. 2018, 3648.
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Sulfonylfluoride (RSO2F)
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Oldest conditions: via RSO2Cl
Harsh conditions
Aryl
Aliphatic
Destabilized and nucleophilic F-
Sharpless, K. et al. Angew. Chem. Int. Ed. 2014, 53, 9430.
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12
Sulfonylfluoride (RSO2F)
Via sulfonate salt
Sun, B.; Qin, H. L. et al. RSC Adv. 2019, 9, 13863.
Pd catalyzed SO2 insertion
Bagley, S. W.; Willis, M. C. et al. Chem. Sci. 2017, 8, 1233.
Ball, N. D. et al. J. Org. Chem. 2017, 82, 2294.
- transition metal free
- one-pot
via:
sulfinate ammonium
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Sulfonylfluoride (RSO2F)
Electrochemical synthesis
No external oxidants
Mild conditions
Safe and accessible F
Noël, T. et al. J. Am. Chem. Soc. 2019, 141, 11832.
Mechanism
Investigation of active species
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SuFEx Hubs
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ESF: ethene sulfonyl fluoride
- Michael addition
- Heck-Matsuda reaction: access to β-aryl ESF
Wu, P.; Sharpless, K. B. et al. Angew. Chem. Int. Ed. 2016, 55, 14155.
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15
SuFEx Hubs- Oxidative Heck reaction
Qin, H. L.; Shalpless, K. B. et al. Angew. Chem., Int. Ed., 2017, 56, 4849.
Arvidsson, P. I. et al. Org. Lett., 2017, 19, 480.
Qin, H. L. et al. Adv. Synth. Catal., 2017, 359, 3237.
- Pd catalyzed coupling with Ar-I
- C-H activation
Org. Chem. Front., 2018, 5, 1411.
Eur. J. Org. Chem., 2018, 4407.
Org. Lett., 2018, 20, 4699.
Organometallics, 2019, 38, 76.
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16Liao, S. et al. Nat. Commun. 2019, 10, 1.
via:
Selected scopes
Radical addition to ESF
Eosin Y-Na2
SuFEx Hubs
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BESF: 1-bromoethene-1-sulfonyl fluoride
Construction of heterocycles
SuFEx Hubs
- Suzuki-Miyaura cross coupling: access to α-aryl ESF
Qin, H. L. et al. Eur. J. Org. Chem. 2019, 6101.
Moses, J. E. et al. Chem. Commun., 2018, 54, 6020.
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Synthesis of Fluorosulfate (ROSO2F)
18Sharpless, K. et al. Angew. Chem. Int. Ed. 2014, 53, 9430.
Reaction with alcohol/phenol
Aryl fluorosulfate is useful in terms of stability.
fluorosulfate
stable for
- months (neutral buffer)
- 2 weeks (buffer pH 10)
Selected scopes
:for simple molecules
:for complex molecules
Sulfuryl fluoride (SO2F2): gas
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Ex Situ Generation of SO2F2 Gas
19De Borggraeve, W. M. et al. Org. Lett. 2017, 19, 5244.
Precursor of SO2F2
27 examples, 85-99%
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20
SO2F2 Equivalents
Sharpless, K. B.; Dong, J. et al. Angew. Chem. Int. Ed. 2018, 57, 2605.
selectivity:
amine, aniline > phenol
sulfamoyl
fluoride
fluorosulfate
Fluorosulfuryl imidazolium salt
Am Ende, C. W. et al. Org. Lett. 2018, 20, 812.
AISF: [(acetylamino)phenyl]imidodisulfuryl difluoride
- bench-stable solid
- higher reactivity than SO2F2
- commercially available from Aldrich- applicable to 1° amine
- no need of SO2F2 for preparation
- bench-stable solid
- higher reactivity than SO2F2
- rapid kinetics
selectivity:
phenol > aniline
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21
Sulfonimidoyl Fluoride (R-SONHF)
Thionyl tetrafluoride (SOF4) gas
orthogonal reactivity
SO2F2: phenol > amine
SOF4: amine > phenol
multidimensional hub
1st addition
Selectivity of nucleophile
Selected scopes
Sharpless, K. B. et al. Angew. Chem. Int. Ed. 2017, 56, 2903.
- 1° amine
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22
Sulfonimidoyl Fluoride (R-SONHF)
2nd addition
- 2° amine
- Amino acid: hydrolytic loss of second S-F
- Aryl silyl ether
sulfonimidoyl fluoride
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23
Sulfonimidoyl Fluoride (R-N=S(O)F2)
Comparison of reactivity
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24
Sulfonimidoyl Fluoride (R-SONHF)
3rd addition
- 2° amine
- Aryl silyl ether
- One-pot procedure
BEMP as potent base to activate inert S-F bond
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25
ApplicationMedicinal chemistry
Chemical biology
New synthetic methodology
Material science
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RSO2F in Biological Context
26
- Activation of S-F in special environment
- Proximity driven reaction
- Ser, Tyr, Lys, Thr, His, Cys
Examples
- Covalent drugs
- Target identification and validation
- Mapping of enzyme binding sites, substrates and protein–protein interactions.
- Late-stage functionalization (LSF) of bioactive molecules
H+ mediated switch-on
-> specificity to microenvironment
electrophilic probe to modify proteins
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Inertness of Sulfonyl Fluoride
27
Conditions: electrophiles (1.0 mmol), aniline 1.3 mL (neat), 184 °C
Nucleophilic stability of electrophiles in chemical biology
Sharpless, K. B. et al. Proc. Natl. Acad. Sci. U. S. A. 2019, 116, 18808.
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Application to Bioconjugation
Modification of BSA with SO2F2 gas
Averick, S. et al. Polymer 2016, 99, 7.
BSA
~4 modifications on Lys
Modification of BSA with fluorosulfate
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SuFEx with Protein in Vivo
Fluorosulfate-L-tyrosine (FSY)
Wang, L. et al. J. Am. Chem. Soc. 2018, 140, 4995.
- FSY was successfully introduced in the protein of E. coli and HELA
- No toxicity to cells during the expression of FSY incorporated proteins.
Incorporation of FSY
Pyrrolysyl-tRNA synthetase (PylRS) mutated library
(mutation at A302, L305, Y306, L309, I322, N346,
C348, Y384, V401, W417)
selection
6 hits PylRS mutant specific for FSY
(mutation 302I, 346T, 348I, 384L, 417K)
-> FSYRS (fluorosulfate tyrosyl-tRNA synthetase)
Coexpression of target protein gene
containing TAG codon with tRNAPylCUA/FSYRS
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30
Afb-Z protein
59% 53% 35%
SuFEx with Protein in Vivo
Western
blot
His tag
purification
-> SDS-PAGE
yieldMBP-Z24FSY, Afb-7X were coexpressed in E. coli.
LC-MS/MS
Croslinking between FSY and Lys/His/Thr
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31
SuFEx with Protein in Vivo
Calmodulin Trx1-PAPS reductase
intramolecular crosslinking crosslinking with native Tyr
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SuFEx Cross-linking for Mass Spectrometry
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Cross-linking mass spectrometry (CXMS)
Identification of protein interaction partners
“plant-and-cast” strategy
Wang, L. et al. Proc. Natl. Acad. Sci. U. S. A. 2018, 115, 11162.
identification of interacting
proteins
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SuFEx Cross-linking for Mass Spectrometry
33
GST
15 crosslinkings
9 crosslinkings
Reaction with peptide
13 crosslinkings
18 crosslinkings
CXMS analysis of BSA
Non-specific cross-linking is observed with BS2G.
Cα-Cα distance (in the crystal structure)
> side chain length*2 + linker length ~ 20 Å
Cross-linking occurred at different structure
from crystal structure.
(in crystal structure)
=
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SuFEx Cross-linking for Mass Spectrometry
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Analysis of Enzyme-Substrate Interactiongenetically encode chemical crosslinkers (GECX)
Additionaly identified Trx-interacting peptides
thioredoxin (Trx)
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Tyr Bioconjugation
35Lee, Y.; Kim, B. M. et al. Chem. Eur. J. 2018, 24, 10948.
(Fs=SO2F)
SuFExMechanism of SuFEx proposed by authors
Deprotection of aryl silyl ether
Use of stoichiometric amount of base avoids the silyl ether?
protein
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36
Tyr Bioconjugation
Residue selectivity Recombinant human erythropoietin (rhEPO)
Screening of base
(5 eq)
PEG
Mn=2372
Δm/z=1980
no silyl activation
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37
Tyr-Reactive Probe
Sulfur-Triazole EXchange (SuTEx)
Hsu, K. et al. Nat. Chem. Biol. doi:10.1038/s41589-019-0404-5
Probes
Proteome analysis
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Tyr-Reactive Probe
Tuning of LG for tyrosine selectivity
Residue selectivity
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Covalent Inhibitor by SuFEx
39
Inhibitor of intracellular lipid binding protein (iLBP)
Sulfonyl fluoride: highly reactivive, resulting in non-specific reactivity
HeLa cell + blank/1/ 2 or S1 (40 uM, 37 °C, 16 h),
CuAAC with cell lysate to attach fluolophore,
SDS-PAGE
Sharpless, K. B.; Kelly, J. W. et al. J. Am. Chem. Soc. 2016, 138, 7353.
Evaluation of proteome reactivity of probes
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40
Stable isotope labeling by amino acids in cell culture (SILAC)
Probe1:
fatty acid binding protein (a family of iLBPs)
FABP 3, FABP 5,
(CRABP2) selective labeling
Probe2:
non-specific labeling
Covalent Inhibitor by SuFEx
HeLa cells13C, 15N12C, 14N
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41
apo CRABP2 (green), FABP5 (magenta), FABP4 (cyan)
Conserved Arg~Arg~Tyr module in iLBPs
Covalent Inhibitor by SuFEx
LC-MS/MS to determine reactive residue
All reacted at Tyr in
Arg~Arg~Tyr module.
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Introduction of biphenyl substructure
Improved reactivity for CRABP2
No reactivity with FABP3, 4, 5
Covalent Inhibitor by SuFEx
Mechanism analysis of selectivity for CRABP2
- Buffer pH dependence
Tyr134 pKa ~ 7.6
effect of Arg111, Arg132?
- Mutation experiment
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crystal structure
large hydrophobic binding
pocket to accommodate
Phenyl ring/ trimethyl
cyclohexene ring
(selectivity of 4 to CRABP2)
retinoic acid (RA)-CRABP2
(PDB: 2FR3)4-CRABP2
Covalent Inhibitor by SuFEx
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PNAS 2019 116 18808との関係が重要。
Covalent inhibitors of human neutrophil elastase (hNE).
Sharpless, K. B. et al. Proc. Natl. Acad. Sci. U. S. A. 2019, 116, 18808.
Covalent Inhibitor by SuFEx
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105 compounds
Covalent Inhibitor by SuFExCategolized SuFExable library
28.5
1
14.1
4.1
relative rate
with ArOTBS
Incubation with enzyme for 10 min
Addition of substrate peptide
(MeOSuc-AAPV-AMC)
Measure increase in fluorescence
1st screen
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Covalent Inhibitor by SuFEx
Crystal structure hNE+1
Hit compounds 2nd screen
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Covalent Inhibitor by SuFEx
Selectivity between serine proteases Effect of denaturation
MALDI-TOF-MS
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Summary
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- Advocated by Sharpless in 2014.
- Stable S(VI)-F bond
- Context dependent activation
Challenges
Mild/sharp activation of S-F bond
Tuning of leaving group instead of F-
Elucidation of mechanism
New synthetic methodology
Proteome analysis
Analysis of PPI
Medicinal chemistry
Polymer synthesis
SuFEx
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Appendix
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Unstable Sulfonyl Fluoride
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PMSF: serine protease inhibitor
short half-life under aqueous conditions
pH 7: 110 min
pH 7.5: 55 min
pH 8: 35 min
unstable under basic conditions
(forms sulfene via E1cB elimination)
Aliphatic sulfonyl fluoride encounter the risk of hydrolysis
under basic conditions.
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Stability under Electrolytic Conditions
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Electrochemical instability of 1,2,3-triazole is demonstrated with CV.
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Radical Addition to ESF
52Liao, S. et al. Nat. Commun. 2019, 10, 1.
Entire mechanism
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Sulfonylfluoride (RSO2F)
Cyclic alkenylsulfonyl fluoride
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RSO2F as Synthetic Reagent
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PyFluor: deoxyfluorination Coupling reagent
JACS 2015, 137, 9571
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RSO2F as Synthetic Reagent
ArOSO2F as cross-coupling partner
✔ high reactivity
-OSO2F ~ -OTf, -Br > -OTs, -OMsby Hanley
-Br ≥ -OTf > -OSO2F > -Clby Sharpless
toxitic
Additive free Suzuki-Miyaura coupling
SO2F2, AISF, imidazolium salt
can be alternative.
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RSO2F as Synthetic Reagent
1,1-Dihydrofluoroalkylation of Amines
- no reaction with –I, -OTs, -OMs
Synthesis of sulfonyl azideSynlett 2016, 27, 1840
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RSO2F as Synthetic Reagent
Diazotizing reagent: primary amine/sulfonamide to azide/sulfonylazide
diazotransfer is reversible
> 40 examples
orthogonal to most of functional groups(including 2°/3° amines)
Sharpless, K. B.; Dong, J. et al. Nature 2019, 574, 86.
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RSO2F as Synthetic Reagent
Sharpless, K. B.; Dong, J. et al. Nature 2019, 574, 86.
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Synthesis of Sulfotyrosine
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Tyr O-sulfation: seen in PTMs
ACIE 2016, 55, 1835
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stable at pH 6-8, 37 °C, 48 h,
hydrolyzed in other range of pH
ACIE 2019, 58, 8029.
iminosulfur oxidifluoride
80–98%
88-97%
95-99% for aryl R1
19% for aliphatic R1
stable at pH 2-14, 37 °C, 96 h
Derivatization under aqueous conditions
Iminosulfur Oxidifluoride under Aqueous Conditions
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DNA modification
Protein modification
Iminosulfur Oxidifluoride under Aqueous Conditions
~30%