• A Corrigendum to this article was published on 18 July 2012

This article has been updated

Abstract

We devised a high-throughput chemoproteomics method that enabled multiplexed screening of 16,000 compounds against native protein and lipid kinases in cell extracts. Optimization of one chemical series resulted in CZC24832, which is to our knowledge the first selective inhibitor of phosphoinositide 3-kinase γ (PI3Kγ) with efficacy in in vitro and in vivo models of inflammation. Extensive target- and cell-based profiling of CZC24832 revealed regulation of interleukin-17–producing T helper cell (TH17) differentiation by PI3Kγ, thus reinforcing selective inhibition of PI3Kγ as a potential treatment for inflammatory and autoimmune diseases.

  • Compound C19H16N6O3S

    N-(6-(5-(Phenylsulfonamido)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C19H16N6O3S

    N-(6-(5-(N-Phenylsulfamoyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C17H20N6O3S

    N-(6-(5-(Butylsulfonamido)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C17H20N6O3S

    N-(6-(5-(N-Butylsulfamoyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C20H15F3N6O4S

    N-(6-(5-(4-(Trifluoromethoxy)phenylsulfonamido)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C20H15F3N6O4S

    N-(6-(5-(N-(4-(Trifluoromethoxy)phenyl)sulfamoyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C19H15ClN6O3S

    N-(6-(5-(4-Chlorophenylsulfonamido)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C19H15ClN6O3S

    N-(6-(5-(N-(4-Chlorophenyl)sulfamoyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C18H13F3N6O2S

    N-(5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridin-3-yl)-3-(trifluoromethyl)benzenesulfonamide

  • Compound C18H13F3N6O2S

    5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-(3-(trifluoromethyl)phenyl)pyridine-3-sulfonamide

  • Compound C18H13F3N6O2S

    N-(5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridin-3-yl)-2-(trifluoromethyl)benzenesulfonamide

  • Compound C18H13F3N6O2S

    5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-(2-(trifluoromethyl)phenyl)pyridine-3-sulfonamide

  • Compound C18H13F3N6O2S

    N-(5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridin-3-yl)-4-(trifluoromethyl)benzenesulfonamide

  • Compound C18H13F3N6O2S

    5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-(4-(trifluoromethyl)phenyl)pyridine-3-sulfonamide

  • Compound C20H20N6O2S

    N-(5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridin-3-yl)-4-isopropylbenzenesulfonamide

  • Compound C20H20N6O2S

    5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-(4-isopropylphenyl)pyridine-3-sulfonamide

  • Compound C17H13ClN6O2S

    N-(5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridin-3-yl)-4-chlorobenzenesulfonamide

  • Compound C17H13ClN6O2S

    5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-(4-chlorophenyl)pyridine-3-sulfonamide

  • Compound C19H12F6N6O2S

    N-(5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridin-3-yl)-3,5-bis(trifluoromethyl)benzenesulfonamide

  • Compound C19H12F6N6O2S

    5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-(3,5-bis(trifluoromethyl)phenyl)pyridine-3-sulfonamide

  • Compound C14H14N4O2

    6-(3,4-Dimethoxyphenyl)-[1,2,4]triazolo[1,5-a]pyridin-2-amine

  • Compound C14H13FN4O2

    6-(3,4-Dimethoxyphenyl)-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-2-amine

  • Compound C15H18N6O2S

    5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-tert-butylpyridine-3-sulfonamide

  • Compound C15H17FN6O2S

    5-(2-Amino-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-tert-butylpyridine-3-sulfonamide

  • Compound C12H12N6O2S

    5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-methylpyridine-3-sulfonamide

  • Compound C12H11FN6O2S

    5-(2-Amino-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-methylpyridine-3-sulfonamide

  • Compound C14H16N6O2S

    5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-isopropylpyridine-3-sulfonamide

  • Compound C14H15FN6O2S

    5-(2-Amino-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-isopropylpyridine-3-sulfonamide

  • Compound C14H16N6O2S

    5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-propylpyridine-3-sulfonamide

  • Compound C14H15FN6O2S

    5-(2-Amino-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-propylpyridine-3-sulfonamide

  • Compound C14H14N6O2S

    6-(5-(Azetidin-1-ylsulfonyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-amine

  • Compound C14H13FN6O2S

    6-(5-(Azetidin-1-ylsulfonyl)pyridin-3-yl)-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-2-amine

  • Compound C13H11F3N6O2S

    5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-(2,2,2-trifluoroethyl)pyridine-3-sulfonamide

  • Compound C13H10F4N6O2S

    5-(2-Amino-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-(2,2,2-trifluoroethyl)pyridine-3-sulfonamide

  • Compound C14H16N6O2S

    5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-ethyl-N-methylpyridine-3-sulfonamide

  • Compound C14H15FN6O2S

    5-(2-Amino-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-ethyl-N-methylpyridine-3-sulfonamide

  • Compound C12H8F3N5

    6-(5-(Trifluoromethyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-amine

  • Compound C12H7F4N5

    8-Fluoro-6-(5-(trifluoromethyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-amine

  • Compound C13H14N6O2S

    5-(2-Amino-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-ethylpyridine-3-sulfonamide

  • Compound C13H13FN6O2S

    5-(2-Amino-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-ethylpyridine-3-sulfonamide

  • Compound C16H16N4O3

    N-(6-(3,4-Dimethoxyphenyl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C16H15FN4O3

    N-(6-(3,4-Dimethoxyphenyl)-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C13H11N5O

    N-(6-(Pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C13H10FN5O

    N-(8-Fluoro-6-(pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C14H13N5O3S

    N-(6-(5-(Methylsulfonyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C14H12FN5O3S

    N-(8-Fluoro-6-(5-(methylsulfonyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide

  • Compound C23H27N9O4S

    N-tert-Butyl-5-(2-(3-(2-(5-cyclopropyl-1,3,4-oxadiazol-2-yl)ethyl)ureido)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridine-3-sulfonamide

  • Compound C23H26FN9O4S

    N-tert-Butyl-5-(2-(3-(2-(5-cyclopropyl-1,3,4-oxadiazol-2-yl)ethyl)ureido)-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridine-3-sulfonamide

  • Compound C20H26N8O4S

    2-(3-(6-(5-(N-tert-Butylsulfamoyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)ureido)-N,N-dimethylacetamide

  • Compound C20H25FN8O4S

    2-(3-(6-(5-(N-tert-Butylsulfamoyl)pyridin-3-yl)-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-2-yl)ureido)-N,N-dimethylacetamide

  • Compound C22H28N8O4S

    N-tert-Butyl-5-(2-(3-(2-oxo-2-(pyrrolidin-1-yl)ethyl)ureido)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridine-3-sulfonamide

  • Compound C22H27FN8O4S

    N-tert-Butyl-5-(8-fluoro-2-(3-(2-oxo-2-(pyrrolidin-1-yl)ethyl)ureido)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridine-3-sulfonamide

  • Compound C22H28N8O5S

    N-tert-Butyl-5-(2-(3-(2-morpholino-2-oxoethyl)ureido)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridine-3-sulfonamide

  • Compound C22H27FN8O5S

    N-tert-Butyl-5-(8-fluoro-2-(3-(2-morpholino-2-oxoethyl)ureido)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridine-3-sulfonamide

  • Compound C23H29N11O3S

    N-tert-Butyl-5-(2-(3-(3-(5-cyclopropyl-2H-tetrazol-2-yl)propyl)ureido)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridine-3-sulfonamide

  • Compound C23H28FN11O3S

    N-tert-Butyl-5-(2-(3-(3-(5-cyclopropyl-2H-tetrazol-2-yl)propyl)ureido)-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridine-3-sulfonamide

  • Compound C20H24N10O3S

    5-(2-(3-(2-(1H-1,2,4-Triazol-1-yl)ethyl)ureido)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-tert-butylpyridine-3-sulfonamide

  • Compound C20H23FN10O3S

    5-(2-(3-(2-(1H-1,2,4-Triazol-1-yl)ethyl)ureido)-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-tert-butylpyridine-3-sulfonamide

  • Compound C20H23N9O4S

    N-tert-Butyl-5-(2-(3-((5-methyl-1,3,4-oxadiazol-2-yl)methyl)ureido)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridine-3-sulfonamide

  • Compound C20H22FN9O4S

    N-tert-Butyl-5-(8-fluoro-2-(3-((5-methyl-1,3,4-oxadiazol-2-yl)methyl)ureido)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyridine-3-sulfonamide

  • Compound C21H27N7O5S

    3-(Dimethylamino)-3-oxopropyl 6-(5-(N-tert-butylsulfamoyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-ylcarbamate

  • Compound C21H26FN7O5S

    3-(Dimethylamino)-3-oxopropyl 6-(5-(N-tert-butylsulfamoyl)pyridin-3-yl)-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-2-ylcarbamate

  • Compound C22H29N7O5S

    2-Morpholinoethyl 6-(5-(N-tert-butylsulfamoyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-ylcarbamate

  • Compound C22H28FN7O5S

    2-Morpholinoethyl 6-(5-(N-tert-butylsulfamoyl)pyridin-3-yl)-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-2-ylcarbamate

  • Compound C19H22N10O4S

    2-(2H-Tetrazol-5-yl)ethyl 6-(5-(N-tert-butylsulfamoyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-ylcarbamate

  • Compound C19H21FN10O4S

    2-(2H-Tetrazol-5-yl)ethyl 6-(5-(N-tert-butylsulfamoyl)pyridin-3-yl)-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-2-ylcarbamate

  • Compound C23H32N8O4S

    2-(4-Methylpiperazin-1-yl)ethyl 6-(5-(N-tert-butylsulfamoyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-ylcarbamate

  • Compound C23H31FN8O4S

    2-(4-Methylpiperazin-1-yl)ethyl 6-(5-(N-tert-butylsulfamoyl)pyridin-3-yl)-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-2-ylcarbamate

  • Compound C22H29N7O4S

    1-Methylpiperidin-4-yl 6-(5-(N-tert-butylsulfamoyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-ylcarbamate

  • Compound C22H28FN7O4S

    1-Methylpiperidin-4-yl 6-(5-(N-tert-butylsulfamoyl)pyridin-3-yl)-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-2-ylcarbamate

  • Compound C17H20N6O4S

    Methyl 6-(5-(N-tert-butylsulfamoyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-ylcarbamate

  • Compound C17H19FN6O4S

    Methyl 6-(5-(N-tert-butylsulfamoyl)pyridin-3-yl)-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-2-ylcarbamate

  • Compound C12H11N5O2S

    6-(5-(Methylsulfonyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-amine

  • Compound C17H19N7O4S

    N,N-Dimethyl-2-(3-(6-(5-(methylsulfonyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)ureido)acetamide

  • Compound C20H20N8O4S

    1-(2-(5-Cyclopropyl-1,3,4-oxadiazol-2-yl)ethyl)-3-(6-(5-(methylsulfonyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)urea

  • Compound C19H22N6O4

    2-(3-(6-(3,4-Dimethoxyphenyl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)ureido)-N,N-dimethylacetamide

  • Compound C16H16N4O4

    Methyl 6-(3,4-dimethoxyphenyl)-[1,2,4]triazolo[1,5-a]pyridin-2-ylcarbamate

  • Compound C19H24N8O4S

    2-(3-(6-(5-(N-Isopropylsulfamoyl)pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)ureido)-N,N-dimethylacetamide

  • Compound C22H32ClN3O7S2

    1-Amino-N-(5-(4-chloro-3-(methylsulfonyl)phenyl)-4-methylthiazol-2-yl)-3,6,9,12-tetraoxapentadecan-15-amide

  • Compound C20H20N2O3

    8-(4-(Aminomethyl)phenyl)-2-morpholino-4H-chromen-4-one

  • Compound C27H26N8O3

    6-((2-Aminoethyl)amino)-N-(3-(4-morpholinopyrido[3',2':4,5]furo[3,2-d]pyrimidin-2-yl)phenyl)nicotinamide

  • Compound C19H17NO3

    2-Morpholino-8-phenyl-4H-chromen-4-one

  • Compound C12H7N3O2S

    (Z)-5-(Quinoxalin-6-ylmethylene)thiazolidine-2,4-dione

  • Compound C14H16ClN3O4S2

    N-(5-(4-Chloro-3-(N-(2-hydroxyethyl)sulfamoyl)phenyl)-4-methylthiazol-2-yl)acetamide

  • Compound C19H16N4O3

    3-(4-Morpholinopyrido[3',2':4,5]furo[3,2-d]pyrimidin-2-yl)phenol

  • Compound C30H23N5O

    2-Methyl-2-(4-(3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)phenyl)propanenitrile

  • Compound C14H8FNO4S

    (Z)-5-((5-(4-Fluoro-2-hydroxyphenyl)furan-2-yl)methylene)thiazolidine-2,4-dione

  • Compound C21H24N4O2

    7-Methyl-2-morpholino-9-(1-(phenylamino)ethyl)-4H-pyrido[1,2-a]pyrimidin-4-one

  • Compound C22H19N7O

    2-((6-Amino-9H-purin-9-yl)methyl)-5-methyl-3-o-tolylquinazolin-4(3H)-one

  • Compound C23H27N7O3S2

    4-(2-(1H-Indazol-4-yl)-6-((4-(methylsulfonyl)piperazin-1-yl)methyl)thieno[3,2-d]pyrimidin-4-yl)morpholine

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  • 16 May 2012

    In the version of this article initially published, the name M. Sunose was misspelled in the Acknowledgements. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We are grateful to M. Sunhose and W. Miller for chemistry and cheminformatics support; MS, information technology, biochemistry and biology groups for technical support; G. Creighton-Gutteridge for experimental support; F. Weisbrodt for assistance with graphics; G. Bennett (Cellzome Ltd.) and O. Azzolino (University of Torino) for animal data; Y. Abraham for generation of the cladogram; R. Hale and T. Edwards for general advice; and M. Wymann, D. Simmons and A. Watt for stimulating discussions and valuable comments.

Author information

Affiliations

  1. Cellzome AG, Heidelberg, Germany.

    • Giovanna Bergamini
    • , Satoko Shimamura
    • , Thilo Werner
    • , Katrin Müller
    • , Jessica Perrin
    • , Christina Rau
    • , Carsten Hopf
    • , Carola Doce
    • , Toby Mathieson
    • , Faiza Rharbaoui
    • , Friedrich Reinhard
    • , Mikhail M Savitski
    • , Gerard Drewes
    • , Marcus Bantscheff
    •  & Gitte Neubauer
  2. Cellzome Ltd., Chesterford Research Park, Cambridge, UK.

    • Kathryn Bell
    • , Andrew Cansfield
    • , Katie Ellard
    • , Daniel Leggate
    • , Raffaella Mangano
    • , Nigel Ramsden
    •  & Oliver Rausch
  3. BioSeek LLC, San Francisco, California, USA.

    • Alison O'Mahony
    •  & Ivan Plavec
  4. Department of Genetics, Biology and Biochemistry, Molecular Biotechnology Center, University of Torino, Torino, Italy.

    • Emilio Hirsch

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Contributions

G.B., S.S., K.M. and J.P. developed and performed assays; K.B., A.C. and K.E. designed and synthesized compounds; C.R., F. Reinhard, D.L. and R.M. performed screening; F. Rharbaoui contributed to animal studies; M.M.S., T.M. and M.B. developed MS technology; C.D. supported data analysis and display; A.O. and I.P. contributed cellular activity profiles; T.W. designed and performed MS experiments; C.H. supervised biochemistry work; G.D. contributed to the manuscript and gave advice; E.H. gave general advice and contributed animal data; N.R. led the chemistry efforts; O.R. contributed to data analysis and manuscript preparation; and G.B., M.B. and G.N. designed and supervised the study, analyzed data and wrote the manuscript.

Competing interests

G.B., S.S., T.W., K.M., J.P., C.R., A.H., C.D., T.M., F. Rharbaoui, F. Reinhard, M.M.S., G.D., M.B. and G.N. are employees of Cellzome AG; K.B., A.C., K.E., D.L., R.M., N.R. and O.R. are employees of Cellzome Ltd.; and A.M. and I.P. are employees of BioSeek Inc.

Corresponding authors

Correspondence to Marcus Bantscheff or Gitte Neubauer.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Methods and Supplementary Results

Excel files

  1. 1.

    Supplementary Data Set 1

    Complementary kinase capturing of CZC15098 and CZC15292 derived bead matrices in different cell extracts

  2. 2.

    Supplementary Data Set 2

    Concentration-inhibition profiles for all reference compounds and CZC19945 and CZC24832 in different cell extracts

  3. 3.

    Supplementary Data Set 3

    Correction factors for calculation of apparent dissociation constants from IC50s obtained in profiling experiments using kinobeads and HL60 cell extracts

  4. 4.

    Supplementary Data Set 4

    Correction factors for calculation of apparent dissociation constants from IC50s obtained in profiling experiments using LK beads and HL60 cell extracts

  5. 5.

    Supplementary Data Set 5

    Proteomic binding profiles using an immobilized linkable analogue of CZC19945/CZC24832 as probe matrix and HeLa, HL60, and PBMC cell extracts

  6. 6.

    Supplementary Data Set 6

    Correction factors for calculation of apparent dissociation constants from IC50s obtained in profiling experiments using LK beads and mixed raw264.7, HL60 cell extracts

  7. 7.

    Supplementary Data Set 7

    Correction factors for calculation of apparent dissociation constants from IC50s obtained in profiling experiments using LK beads and mixed RBL-1, HL60 cell extracts

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DOI

https://doi.org/10.1038/nchembio.957

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