DOI: 10.1126/science.276.5314.955. Structures of the Tyrosine Kinase Domain of Fibroblast Growth Factor Receptor in Complex with Inhibitors

Structures of the Tyrosine Kinase Domain of Fibroblast Growth Factor Receptor in Complex with Inhibitors

10.1126/science.276.5314.955

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

A new class of protein tyrosine kinase inhibitors was identified that is based on an oxindole core (indolinones). Two compounds from this class inhibited the kinase activity of fibroblast growth factor receptor 1 (FGFR1) and showed differential specificity toward other receptor tyrosine kinases. Crystal structures of the tyrosine kinase domain of FGFR1 in complex with the two compounds were determined. The oxindole occupies the site in which the adenine of adenosine triphosphate binds, whereas the moieties that extend from the oxindole contact residues in the hinge region between the two kinase lobes. The more specific inhibitor of FGFR1 induces a conformational change in the nucleotide-binding loop. This structural information will facilitate the design of new inhibitors for use in the treatment of cancer and other diseases in which cell signaling by tyrosine kinases plays a crucial role in disease pathogenesis.

Bibliography

Mohammadi, M., McMahon, G., Sun, L., Tang, C., Hirth, P., Yeh, B. K., Hubbard, S. R., & Schlessinger, J. (1997). Structures of the Tyrosine Kinase Domain of Fibroblast Growth Factor Receptor in Complex with Inhibitors. Science, 276(5314), 955â960.

Authors 8

Moosa Mohammadi (first)
Gerald McMahon (additional)
Li Sun (additional)
Cho Tang (additional)
Peter Hirth (additional)
Brian K. Yeh (additional)
Stevan R. Hubbard (additional)
Joseph Schlessinger (additional)

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Preparation of 3-[4-(1-formylpiperazin-4-yl)benzylidenyl]-2-indolinone (SU4984) was as follows: A reaction mixture of 133.15 mg (1.0 mmol) of oxindole 228.3 mg (1.2 mmol) of 4-(1-formylpiperazin-4- yl)benzaldehyde and three drops of piperidine in 2 ml of ethanol was stirred at 90°C for 5 hours. After cooling the precipitate was filtered washed with cold ethanol and dried to yield 199.5 mg (65%) of SU4984 as a yellow solid. Nuclear magnetic resonance (NMR) spectroscopy showed that SU4984 exists predominantly in the trans configuration (C-2 C-1′) although in the crystal structure SU4984 is observed in the cis configuration. Preparation of 3-[(3-(2-carboxyethyl)-4-methylpyrrol-2-yl)methylene]-2-indolinone (SU5402) was as follows: A reaction mixture of 134.0 mg (1.0 mmol) of oxindole 217.43 mg (1.2 mmol) of 3-(2-carboxyethyl)-4-methylpyrrol-2-carboxaldehyde and three drops of piperidine in 3 ml of ethanol was stirred at 90°C for 3 hours. After cooling the precipitate was filtered washed with cold ethanol and dried to yield 172.4 mg (58%) of SU5402 as a yellow solid. NMR spectroscopy showed that SU5402 exists predominantly in the cis configuration.
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Equipment in the structural biology program at the Skirball Institute is partially supported by a grant from the Kresge Foundation. Coordinates have been deposited in the Brookhaven Protein Data Bank entries 1FGI and 1AGW.

Dates

Type	When
Created	23 years, 1 month ago (July 27, 2002, 1:45 a.m.)
Deposited	1 year, 7 months ago (Jan. 12, 2024, 7:46 p.m.)
Indexed	1 day, 12 hours ago (Aug. 26, 2025, 2:30 a.m.)
Issued	28 years, 3 months ago (May 9, 1997)
Published	28 years, 3 months ago (May 9, 1997)
Published Print	28 years, 3 months ago (May 9, 1997)

Funders 0

None

BibTeX

@article{Mohammadi_1997, title={Structures of the Tyrosine Kinase Domain of Fibroblast Growth Factor Receptor in Complex with Inhibitors}, volume={276}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.276.5314.955}, DOI={10.1126/science.276.5314.955}, number={5314}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Mohammadi, Moosa and McMahon, Gerald and Sun, Li and Tang, Cho and Hirth, Peter and Yeh, Brian K. and Hubbard, Stevan R. and Schlessinger, Joseph}, year={1997}, month=may, pages={955–960} }

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      "author": "Webster M. K.",
      "year": "1996",
      "unstructured": "Webster M. K., et al., Mol. Cell. Biol. 16, 4081 (1996).",
      "journal-title": "Mol. Cell. Biol."
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      "doi-asserted-by": "crossref",
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      "DOI": "10.1161/01.ATV.9.3.269",
      "volume": "9",
      "author": "Klagsbrun M.",
      "year": "1989",
      "unstructured": "Klagsbrun M., Edelman E. R., Arteriosclerosis 9, 269 (1989);",
      "journal-title": "Arteriosclerosis"
    },
    {
      "key": "e_1_3_1_10_3",
      "doi-asserted-by": "crossref",
      "unstructured": "; H. Brem and M. Klagsbrun in Oncogenes and Tumor Suppressor Genes in Human Malignancies C. C. Benz and E. T. Liu Eds. (Kluwer Academic Boston 1993) p. 211; M. Klagsbrunand P. A. D\u2019Amore Ann. Rev. Physiol. 53 217 (1991).",
      "DOI": "10.1007/978-1-4615-3088-6_10"
    },
    {
      "key": "e_1_3_1_11_2",
      "first-page": "659",
      "volume": "6",
      "author": "Adnane J.",
      "year": "1991",
      "unstructured": "Adnane J., et al., Oncogene 6, 659 (1991);",
      "journal-title": "Oncogene"
    },
    {
      "key": "e_1_3_1_11_3",
      "doi-asserted-by": "crossref",
      "first-page": "170",
      "DOI": "10.1002/ijc.2910610205",
      "volume": "61",
      "author": "Penault-Llorca F.",
      "year": "1995",
      "unstructured": "Penault-Llorca F., et al., Int. J. Cancer 61, 170 (1995);",
      "journal-title": "Int. J. Cancer"
    },
    {
      "key": "e_1_3_1_11_4",
      "first-page": "373",
      "volume": "59",
      "author": "Jacquemier J.",
      "year": "1994",
      "unstructured": "Jacquemier J., et al., ibid. 59, 373 (1994);",
      "journal-title": "ibid."
    },
    {
      "key": "e_1_3_1_11_5",
      "first-page": "523",
      "volume": "54",
      "author": "McLeskey S. W.",
      "year": "1994",
      "unstructured": "McLeskey S. W., Ding I. Y., Lippman M. E., Kern F. G., Cancer Res. 54, 523 (1994);",
      "journal-title": "Cancer Res."
    },
    {
      "key": "e_1_3_1_11_6",
      "doi-asserted-by": "crossref",
      "first-page": "274",
      "DOI": "10.1002/ijc.2910640411",
      "volume": "64",
      "author": "Luqmani Y. A.",
      "year": "1995",
      "unstructured": "Luqmani Y. A., et al., Int. J. Cancer 64, 274 (1995);",
      "journal-title": "Int. J. Cancer"
    },
    {
      "key": "e_1_3_1_11_7",
      "first-page": "374",
      "volume": "54",
      "author": "Jaakkola S.",
      "year": "1993",
      "unstructured": "Jaakkola S., et al., ibid. 54, 374 (1993).",
      "journal-title": "ibid."
    },
    {
      "key": "e_1_3_1_12_2",
      "doi-asserted-by": "crossref",
      "first-page": "667",
      "DOI": "10.1002/ijc.2910590515",
      "volume": "59",
      "author": "Leung H. Y.",
      "year": "1994",
      "unstructured": "Leung H. Y., Gullick W. J., Lemoine N. R., Int. J. Cancer 59, 667 (1994).",
      "journal-title": "Int. J. Cancer"
    },
    {
      "key": "e_1_3_1_13_2",
      "doi-asserted-by": "crossref",
      "unstructured": "F. Yamaguchi H. Saya J. M. Bruner R. S. Morrison Proc. Natl. Acad. Sci. U.S.A. 91 484 (1994);",
      "DOI": "10.1073/pnas.91.2.484"
    },
    {
      "key": "e_1_3_1_13_3",
      "doi-asserted-by": "crossref",
      "first-page": "207",
      "DOI": "10.1007/BF01328955",
      "volume": "18",
      "author": "Morrison R. S.",
      "year": "1994",
      "unstructured": "Morrison R. S., et al., J. Neurooncol. 18, 207 (1994).",
      "journal-title": "J. Neurooncol."
    },
    {
      "key": "e_1_3_1_14_2",
      "doi-asserted-by": "crossref",
      "first-page": "650",
      "DOI": "10.1002/(SICI)1097-0215(19960301)65:5<650::AID-IJC15>3.0.CO;2-B",
      "volume": "65",
      "author": "Myoken Y.",
      "year": "1996",
      "unstructured": "Myoken Y., et al., Int. J. Cancer 65, 650 (1996).",
      "journal-title": "Int. J. Cancer"
    },
    {
      "key": "e_1_3_1_15_2",
      "doi-asserted-by": "crossref",
      "first-page": "2253",
      "DOI": "10.1002/1097-0142(19931001)72:7<2253::AID-CNCR2820720732>3.0.CO;2-4",
      "volume": "72",
      "author": "Li J. J.",
      "year": "1993",
      "unstructured": "Li J. J., et al., Cancer 72, 2253 (1993).",
      "journal-title": "Cancer"
    },
    {
      "key": "e_1_3_1_16_2",
      "doi-asserted-by": "crossref",
      "first-page": "219",
      "DOI": "10.1002/gcc.2870070407",
      "volume": "7",
      "author": "Theillet C.",
      "year": "1993",
      "unstructured": "Theillet C., et al., Genes Chromosomes Cancer 7, 219 (1993).",
      "journal-title": "Genes Chromosomes Cancer"
    },
    {
      "key": "e_1_3_1_17_2",
      "doi-asserted-by": "crossref",
      "first-page": "297",
      "DOI": "10.1007/BF00185973",
      "volume": "13",
      "author": "Story M. T.",
      "year": "1995",
      "unstructured": "Story M. T., World J. Urol. 13, 297 (1995);",
      "journal-title": "World J. Urol."
    },
    {
      "key": "e_1_3_1_17_3",
      "doi-asserted-by": "crossref",
      "first-page": "287",
      "DOI": "10.1007/BF00665959",
      "volume": "12",
      "author": "Ware J. L.",
      "year": "1993",
      "unstructured": "Ware J. L., Cancer Metastasis Rev. 12, 287 (1993).",
      "journal-title": "Cancer Metastasis Rev."
    },
    {
      "key": "e_1_3_1_18_2",
      "unstructured": "Preparation of 3-[4-(1-formylpiperazin-4-yl)benzylidenyl]-2-indolinone (SU4984) was as follows: A reaction mixture of 133.15 mg (1.0 mmol) of oxindole 228.3 mg (1.2 mmol) of 4-(1-formylpiperazin-4- yl)benzaldehyde and three drops of piperidine in 2 ml of ethanol was stirred at 90\u00b0C for 5 hours. After cooling the precipitate was filtered washed with cold ethanol and dried to yield 199.5 mg (65%) of SU4984 as a yellow solid. Nuclear magnetic resonance (NMR) spectroscopy showed that SU4984 exists predominantly in the trans configuration (C-2 C-1\u2032) although in the crystal structure SU4984 is observed in the cis configuration. Preparation of 3-[(3-(2-carboxyethyl)-4-methylpyrrol-2-yl)methylene]-2-indolinone (SU5402) was as follows: A reaction mixture of 134.0 mg (1.0 mmol) of oxindole 217.43 mg (1.2 mmol) of 3-(2-carboxyethyl)-4-methylpyrrol-2-carboxaldehyde and three drops of piperidine in 3 ml of ethanol was stirred at 90\u00b0C for 3 hours. After cooling the precipitate was filtered washed with cold ethanol and dried to yield 172.4 mg (58%) of SU5402 as a yellow solid. NMR spectroscopy showed that SU5402 exists predominantly in the cis configuration."
    },
    {
      "key": "e_1_3_1_19_2",
      "unstructured": "The compounds were dissolved in dimethylsulfoxide (DMSO) as 100 mM stock solutions and stored at 4\u00b0C. The compounds were then diluted in DMSO to a concentration 20\u00d7 the final concentration used in the in vitro kinase reactions. FGFR1K (9 \u03bcl) [2.2 mg/ml in 10 mM tris (pH 8) and 10 mM NaCl] was mixed with 1 \u03bcl of various concentrations of 20\u00d7 SU4984 or 20\u00d7 SU5402 or with 1 \u03bcl of DMSO (control). The reaction was started by adding 6 \u03bcl of the enzyme-compound mixture to 6 \u03bcl of 2\u00d7 kinase buffer {2 mM ATP/[\u03b3- 32 P]ATP (10 \u03bcCi/\u03bcl) 4 mM MgCl 2 in 10 mM tris (pH 8) and 10 mM NaCl} at room temperature. At various time points 2 \u03bcl of the reaction mixture was removed and added to 4 \u03bcl of 20 mM EDTA to stop the reaction. The reaction products were analyzed by SDS-PAGE (12% gel) and autoradiography. The radioactive bands were excised from the gel and 32 P incorporation was quantitated by Cerenkov counting with a \u03b2 counter."
    },
    {
      "key": "e_1_3_1_20_2",
      "unstructured": "NIH 3T3 cells expressing endogenous FGF receptors were used. Cell culture cell lysis immunoprecipitation and immunoblotting were carried out according to standard procedures. Anti-FGFR1 was described previously ["
    },
    {
      "key": "e_1_3_1_20_3",
      "doi-asserted-by": "crossref",
      "first-page": "977",
      "DOI": "10.1128/MCB.16.3.977",
      "volume": "16",
      "author": "Mohammadi M.",
      "year": "1996",
      "unstructured": "Mohammadi M., et al., Mol. Cell. Biol. 16, 977 (1996)].",
      "journal-title": "Mol. Cell. Biol."
    },
    {
      "key": "e_1_3_1_21_2",
      "unstructured": "[ 3 H]Thymidine incorporation was done as described by Mohammadi et al. (19)."
    },
    {
      "key": "e_1_3_1_22_2",
      "unstructured": "NIH 3T3 cells expressing endogenous PDGF receptors were used. Also used were HER14 cells (33) and NIHIR cells ["
    },
    {
      "key": "e_1_3_1_22_3",
      "doi-asserted-by": "crossref",
      "first-page": "3959",
      "DOI": "10.1074/jbc.271.8.3959",
      "volume": "271",
      "author": "Isakoff S. J.",
      "year": "1996",
      "unstructured": "Isakoff S. J., et al., J. Biol. Chem. 271, 3959 (1996);",
      "journal-title": "J. Biol. Chem."
    },
    {
      "key": "e_1_3_1_22_4",
      "unstructured": "]. The EGF receptor was immunoprecipitated with monoclonal antibody 108 (33) insulin receptor with monoclonal antibodies (BBE) ["
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      "doi-asserted-by": "crossref",
      "first-page": "23126",
      "DOI": "10.1074/jbc.270.39.23126",
      "volume": "270",
      "author": "Moller N. P.",
      "year": "1995",
      "unstructured": "Moller N. P., et al., J. Biol. Chem. 270, 23126 (1995);",
      "journal-title": "J. Biol. Chem."
    },
    {
      "key": "e_1_3_1_22_6",
      "unstructured": "] and PDGF receptor with polyclonal antibodies (PR4) ["
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    {
      "key": "e_1_3_1_22_7",
      "doi-asserted-by": "crossref",
      "first-page": "2257",
      "DOI": "10.1002/j.1460-2075.1993.tb05879.x",
      "volume": "12",
      "author": "Mori S.",
      "year": "1993",
      "unstructured": "Mori S., et al., EMBO J. 12, 2257 (1993)].",
      "journal-title": "EMBO J."
    },
    {
      "key": "e_1_3_1_23_2",
      "doi-asserted-by": "crossref",
      "first-page": "577",
      "DOI": "10.1016/S0092-8674(00)80131-2",
      "volume": "86",
      "author": "Mohammadi M.",
      "year": "1996",
      "unstructured": "Mohammadi M., Schlessinger J., Hubbard S. R., Cell 86, 577 (1996).",
      "journal-title": "Cell"
    },
    {
      "key": "e_1_3_1_24_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0969-2126(00)00036-8"
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      "doi-asserted-by": "crossref",
      "first-page": "4843",
      "DOI": "10.1073/pnas.79.16.4843",
      "volume": "79",
      "author": "Thomas K. A.",
      "year": "1982",
      "unstructured": "Thomas K. A., Smith G. M., Thomas T. B., Feldman R. J., Proc. Natl. Acad. Sci. U.S.A. 79, 4843 (1982).",
      "journal-title": "Proc. Natl. Acad. Sci. U.S.A."
    },
    {
      "key": "e_1_3_1_26_2",
      "unstructured": "G. McMahon unpublished results."
    },
    {
      "key": "e_1_3_1_27_2",
      "unstructured": "S. R. Hubbard in preparation."
    },
    {
      "key": "e_1_3_1_28_2",
      "doi-asserted-by": "crossref",
      "first-page": "746",
      "DOI": "10.1038/372746a0",
      "volume": "372",
      "author": "___",
      "year": "1994",
      "unstructured": "___, Wei L., Ellis L., Hendrickson W. A., Nature 372, 746 (1994).",
      "journal-title": "Nature"
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      "doi-asserted-by": "crossref",
      "first-page": "14503",
      "DOI": "10.1016/S0021-9258(18)71707-9",
      "volume": "264",
      "author": "Lyall R. M.",
      "year": "1989",
      "unstructured": "Lyall R. M., et al., J. Biol. Chem. 264, 14503 (1989);",
      "journal-title": "J. Biol. Chem."
    },
    {
      "key": "e_1_3_1_29_3",
      "doi-asserted-by": "crossref",
      "first-page": "1101",
      "DOI": "10.1016/0092-8674(89)90047-0",
      "volume": "57",
      "author": "Margolis B.",
      "year": "1989",
      "unstructured": "Margolis B., et al., Cell 57, 1101 (1989).",
      "journal-title": "Cell"
    },
    {
      "key": "e_1_3_1_30_2",
      "doi-asserted-by": "crossref",
      "first-page": "2735",
      "DOI": "10.1073/pnas.93.7.2735",
      "volume": "93",
      "author": "De Azevedo W. F.",
      "year": "1996",
      "unstructured": "De Azevedo W. F., et al., Proc. Natl. Acad. Sci. U.S.A. 93, 2735 (1996);",
      "journal-title": "Proc. Natl. Acad. Sci. U.S.A."
    },
    {
      "key": "e_1_3_1_30_3",
      "doi-asserted-by": "crossref",
      "first-page": "378",
      "DOI": "10.1002/prot.340220408",
      "volume": "22",
      "author": "Schulze-Gahmen U.",
      "year": "1995",
      "unstructured": "Schulze-Gahmen U., et al., Proteins Struct. Funct. Genet. 22, 378 (1995).",
      "journal-title": "Proteins Struct. Funct. Genet."
    },
    {
      "key": "e_1_3_1_31_2",
      "doi-asserted-by": "crossref",
      "first-page": "6308",
      "DOI": "10.1073/pnas.93.13.6308",
      "volume": "93",
      "author": "Xu R.-M.",
      "year": "1996",
      "unstructured": "Xu R.-M., Carmel G., Kuret J., Cheng X., Proc. Natl. Acad. Sci. U.S.A. 93, 6308 (1996).",
      "journal-title": "Proc. Natl. Acad. Sci. U.S.A."
    },
    {
      "key": "e_1_3_1_32_2",
      "doi-asserted-by": "crossref",
      "first-page": "26157",
      "DOI": "10.1074/jbc.271.42.26157",
      "volume": "271",
      "author": "Engh R. A.",
      "year": "1996",
      "unstructured": "Engh R. A., Girod A., Kinzel V., Huber R., Bossemeyer D., J. Biol. Chem. 271, 26157 (1996).",
      "journal-title": "J. Biol. Chem."
    },
    {
      "key": "e_1_3_1_33_2",
      "unstructured": "M. G. Malkin W. P. Mason F. S. Lieberman L. K. Shawver A. L. Hannah in Proceedings of the 32nd Annual Meeting of the American Society of Clinical Oncology Philadelphia PA 18 to 21 May 1996 (American Society of Clinical Oncology Philadelphia PA 1996) vol. 15 p. 1573."
    },
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      "doi-asserted-by": "crossref",
      "first-page": "199",
      "DOI": "10.1016/0092-8674(87)90147-4",
      "volume": "51",
      "author": "Honegger A. M.",
      "year": "1987",
      "unstructured": "Honegger A. M., et al., Cell 51, 199 (1987).",
      "journal-title": "Cell"
    },
    {
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      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0263-7855(93)87009-T"
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      "doi-asserted-by": "publisher",
      "DOI": "10.1002/prot.340110407"
    },
    {
      "key": "e_1_3_1_37_2",
      "unstructured": "Z. Otwinowski in Data Collection and Processing L. Sawyer N. Isaacs S. Bailey Eds. (SERC Daresbury Laboratory Warrington UK 1993) p. 56."
    },
    {
      "key": "e_1_3_1_38_2",
      "unstructured": "A. T. Br\u00fcnger X-PLOR Version 3.1: A System for X-ray and NMR (Yale Univ. Press New Haven CT 1992)."
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      "doi-asserted-by": "crossref",
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      "DOI": "10.1016/0076-6879(85)15014-7",
      "volume": "115",
      "author": "Jones T. A.",
      "year": "1985",
      "unstructured": "Jones T. A., Methods Enzymol. 115, 157 (1985).",
      "journal-title": "Methods Enzymol."
    },
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      "key": "e_1_3_1_40_2",
      "unstructured": "Equipment in the structural biology program at the Skirball Institute is partially supported by a grant from the Kresge Foundation. Coordinates have been deposited in the Brookhaven Protein Data Bank entries 1FGI and 1AGW."
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