New Cyclic Arg-Gly-Asp Pseudopentapeptide Containing the β-Turn Mimetic GPTM

Maria Salvati; Franca M. Cordero; Federica Pisaneschi; Nicoletta Cini; Anna Bottoncetti; Alberto Brandi

doi:10.1055/s-2006-949610

LETTER

New Cyclic Arg-Gly-Asp Pseudopentapeptide Containing the β-Turn Mimetic GPTM

Maria Salvati^a, Franca M. Cordero*^a, Federica Pisaneschi^a, Nicoletta Cini^b, Anna Bottoncetti^b, Alberto Brandi*^a
^a Department of Organic Chemistry ‘Ugo Schiff’, Laboratory of Design, Synthesis and Study of Biologically Active Heterocycles, University of Firenze, via della Lastruccia 13, 50019 Sesto Fiorentino (FI), Italy
Fax: +39(055)4573531; e-Mail: franca.cordero@unifi.it; e-Mail: alberto.brandi@unifi.it;
^b Department of Clinical Physiopathology, Nuclear Medicine Unit, University of Firenze, viale G. Pieraccini 6, 50134 Firenze, Italy

Abstract

All articles of this category

Abstract

The solid-phase synthesis of cyclic pseudopeptides containing the Arg-Gly-Asp recognition sequence and the dipeptide isostere 2-amino-3-oxotetrahydro-1H-pyrrolizine-7a(5H)-carboxylic acid (GPTM) was accomplished. N-Fmoc-Asp-OAll was anchored to Wang resin through its side chain and sequentially coupled with N-Fmoc-Gly-OH, N-Fmoc-Arg(Pbf)-OH and N-Fmoc-GPTM-OH. The supported linear pentapeptide smoothly underwent head-to-tail cyclization by activation with TBTU/DIPEA. Finally, TFA treatment released the completely deprotected cyclic pseudopentapeptide. Competition binding assays to purified α_Vβ₃ and α_Vβ₅ integrins showed a high inhibitory activity of cyclo[Arg-Gly-Asp-(2S,7aS)-GPTM].

Key words

cyclic peptides - solid-phase synthesis - heterocycles - integrin antagonists - dipeptide isosteres

Full Text

References

References and Notes

For recent reviews, see:

<A NAME="RG16706ST-1A">1a</A> Arnaout MA. Goodman SL. Xiong JP. Curr. Opin. Cell Biol. 2002, 14: 641

<A NAME="RG16706ST-1B">1b</A> Hynes RO. Cell 2002, 110: 673

<A NAME="RG16706ST-1C">1c</A> Plow EF. Haas TA. Zhang L. Loftus J. Smith JW. J. Biol. Chem. 2000, 275: 21785

<A NAME="RG16706ST-1D">1d</A> Humphries MJ. Biochem. Soc. Trans. 2000, 28: 311

<A NAME="RG16706ST-2A">2a</A> Eliceiri BP. Cheresh DA. Cancer J. 2000, 13: 245

<A NAME="RG16706ST-2B">2b</A> Burke PA. De Nardo SJ. Miers LA. Lamborn KL. Matzku S. De Nardo GL. Cancer Res. 2002, 62: 4263

<A NAME="RG16706ST-2C">2c</A> Haubner R. Finsinger D. Kessler H. Angew. Chem., Int. Ed. Engl. 1997, 36: 1374

<A NAME="RG16706ST-3A">3a</A> Marinelli L. Gottschalk KE. Meyer A. Novellino E. Kessler H. J. Med. Chem. 2004, 47: 4166

<A NAME="RG16706ST-3B">3b</A> Friedlander M. Theesfeld CL. Sugita M. Fruttiger M. Thomas MA. Chang S. Cheresh DA. Proc. Natl. Acad. Sci. U.S.A. 1996, 93: 9764

<A NAME="RG16706ST-3C">3c</A> Friedlander M. Brooks PC. Shaffer RW. Kincaid CM. Varner JA. Cheresh DA. Science 1995, 270: 1550

<A NAME="RG16706ST-4">4</A> Ruoslathi E. Pierschbacher MD. Science 1987, 238: 491

<A NAME="RG16706ST-5">5</A> Haubner R. Gratias R. Diefenbach B. Goodman SL. Jonczyk A. Kessler H. J. Am. Chem. Soc. 1996, 118: 7461

<A NAME="RG16706ST-6A">6a</A> Bach AC. Espina JR. Jackson SA. Stouten PFW. Duke JL. Mousa SA. DeGrado WF. J. Am. Chem. Soc. 1996, 118: 293

<A NAME="RG16706ST-6B">6b</A> Dechantsreiter M. Plnaker E. Mathä B. Lohof E. Hölzemann G. Jonczyk A. Goodman SL. Kessler H. J. Med. Chem. 1999, 42: 3033

<A NAME="RG16706ST-6C">6c</A> Lohof E. Plnaker E. Mang C. Burkhart F. Dechantsreiter MA. Haubner R. Wester HJ. Schwaiger M. Hölzemann G. Goodman SL. Kessler H. Angew. Chem. Int. Ed. 2000, 39: 2761

<A NAME="RG16706ST-6D">6d</A> Belvisi L. Bernardi A. Checchia A. Manzoni L. Potenza D. Scolastico C. Castorina M. Capelli A. Giannini G. Carminati P. Pisano C. Org. Lett. 2001, 3: 1001

<A NAME="RG16706ST-6E">6e</A> Belvisi L. Bernardi A. Colombo M. Manzoni L. Potenza D. Scolastico C. Giannini G. Marcellini M. Riccioni T. Castorina M. LoGiudice P. Carminati P. Pisano C. Bioorg. Med. Chem. 2006, 14: 169

<A NAME="RG16706ST-7A">7a</A> Pitts WJ. Wityak J. Smallheer JM. Tobin AE. Jetter JW. Buynitsky JS. Harlow PP. Solomon KA. Corjay MH. Mousa SA. Wexler RR. Jadhav PK. J. Med. Chem. 2000, 43: 27

<A NAME="RG16706ST-7B">7b</A> Batt DG. Petraitis J. Houghton GC. Modi DP. Cain GA. Corjay MH. Mousa SA. Bouchard PJ. Forsythe MS. Harlow PP. Barbera FA. Spitz SM. Wexler RR. Jadhav PK. J. Med. Chem. 2000, 43: 41

<A NAME="RG16706ST-8A">8a</A> Pisaneschi F. Gensini M. Salvati M. Cordero FM. Brandi A. Heterocycles 2006, 67: 413

<A NAME="RG16706ST-8B">8b</A> Salvati M. Cordero FM. Pisaneschi F. Bucelli F. Brandi A. Tetrahedron 2005, 61: 8836

<A NAME="RG16706ST-8C">8c</A> Cordero FM. Pisaneschi F. Salvati M. Valenza S. Faggi C. Brandi A. Chirality 2005, 17: 149

<A NAME="RG16706ST-8D">8d</A> Cordero FM. Pisaneschi F. Meschini Batista K. Valenza S. Machetti F. Brandi A. J. Org. Chem. 2005, 70: 856

<A NAME="RG16706ST-8E">8e</A> Cordero FM. Valenza S. Machetti F. Brandi A. Chem. Commun. 2001, 1590

<A NAME="RG16706ST-9">9</A> For a recent review on azabicyclic dipeptide mimetics, see: Maison W. Prenzel AHGP. Synthesis 2005, 1031

For recent reviews, see:

<A NAME="RG16706ST-10A">10a</A> Davies JS. J. Pept. Sci. 2003, 9: 471

<A NAME="RG16706ST-10B">10b</A> Lambert JN. Mitchell P. Roberts KD. J. Chem. Soc., Perkin Trans. 1 2001, 5: 471

<A NAME="RG16706ST-10C">10c</A> Romanovskis P. Spatola AF. J. Pept. Res. 1998, 52: 356

<A NAME="RG16706ST-11A">11a</A> Mazur S. Jayalekshmy P. J. Am. Chem. Soc. 1979, 101: 677

<A NAME="RG16706ST-11B">11b</A> Albericio F. Hammer RP. García-Echeverría C. Molins MA. Cheng JL. Munson MC. Pons M. Giralt E. Barany G. Int. J. Pept. Protein Res. 1991, 37: 402

<A NAME="RG16706ST-12A">12a</A> Alcaro MC. Sabatino G. Uziel J. Chelli M. Ginanneschi M. Rovero P. Papini AM. J. Pept. Sci. 2004, 10: 218

<A NAME="RG16706ST-12B">12b</A> Chaleix V. Sol V. Guilloton M. Granet R. Krausz P. Tetrahedron Lett. 2004, 45: 5295

<A NAME="RG16706ST-12C">12c</A> Royo M. Farrera-Sinfreu J. Solé L. Albericio F. Tetrahedron Lett. 2002, 43: 2029

<A NAME="RG16706ST-12D">12d</A> McCusker CF. Kocienski PJ. Botle FT. Schätzlein AG. Bioorg. Med. Chem. 2002, 12: 547

<A NAME="RG16706ST-12E">12e</A> Akaji K. Teruya K. Akaji M. Aimoto S. Tetrahedron 2001, 57: 2293

<A NAME="RG16706ST-12F">12f</A> Delforge D. Art M. Gillon B. Die M. Delaive E. Raes M. Remacle J. Anal. Biochem. 1996, 242: 180

<A NAME="RG16706ST-12G">12g</A> Kates SA. Daniels SB. Albericio F. Anal. Biochem. 1993, 212: 303

<A NAME="RG16706ST-13">13</A> Kaiser E. Colescott RL. Bossinger CD. Cook PI. Anal. Biochem. 1970, 34: 595

<A NAME="RG16706ST-14">14</A> Krchnak V. Vagner J. Safar P. Lebl M. Collect. Czech. Chem. Commun. 1988, 53: 2542

Analytical Data for Cyclo[Arg-Gly-Asp-(2S,7aS)-GPTM] (3). Yield of 14% with respect to N-Fmoc-Asp(Wang-resin)-OAll loading; [α]_D ²⁰ +44.0 (c 0.3, H₂O). ¹H NMR (400 MHz, D₂O): δ = 4.10 (t, J = 7.7 Hz, 1 H, H-C_α Arg), 4.05 (d, J = 14.2 Hz, 1 H, H-C_α Gly), 3.42 (d, J = 14.1 Hz, 1 H, H-C_α Gly), 3.34 (dt, J = 11.7, 8.2 Hz, 1 H, 5-H_a GPTM), 3.16-3.02 (m, 3 H, 5-H_b GPTM, H-C_δ Arg), 2.78 (dd, J = 7.6, 6.0 Hz, 2 H, H-C_β Asp), 2.47 (dd, J = 14.3, 8.2 Hz, 1 H, 1-H_a GPTM), 2.29 (dd, J = 12.3, 5.7 Hz, 1 H, 7-H_a GPTM), 2.13 (d, J = 14.3 Hz, 1 H, 1-H_b GPTM), 2.12-2.08 (m, 1 H, 6-H_a GPTM), 1.84-1.77 (m, 1 H, 7-H_b GPTM), 1.70 (q, J = 7.7 Hz, 2 H, H-C_β Arg), 1.62-1.43 (m, 3 H, 6-H_b GPTM, H-C_γ Arg). ¹H NMR (400 MHz, DMSO-d ₆): δ = 12.20 (br s, 1 H, CO₂H), 8.66 (br t, J = 6.1 Hz, 1 H, NH Gly), 8.41 (d, J = 6.8 Hz, 1 H, NH Arg), 8.28 (d, J = 9.0 Hz, 1 H, NH GPTM), 7.44 (br s, 2 H, NH₂ Arg), 7.40-7.38 (m, 1 H, C=NH Arg), 7.13 (d, J = 9.2 Hz, 1 H, NH Asp), 4.56 (dt, J = 8.4, 5.1 Hz, 1 H, 2-H GPTM), 4.47 (dt, J = 8.5, 4.3 Hz, 1 H, H-C_α Asp), 4.07 (q, J = 7.2 Hz, 1 H, H-C_α Arg), 4.00 (dd, J = 13.9, 7.8 Hz, 1 H, H-C_α Gly), 3.48-3.45 (m, 1 H, 5-H_a GPTM), 3.19 (dd, J = 13.9, 4.4 Hz, 1 H, H-C_α Gly), 3.12-3.07 (m, 2 H, H-C_δ Arg), 2.99-2.92 (m, 1 H, 5-H_b GPTM), 2.69 (dd, J = 16.7, 8.4 Hz, 1 H, 1-H_a GPTM), 2.41 (dd, J = 16.7, 4.8 Hz, 1 H, 1-H_b GPTM), 2.34-2.29 (m, 3 H, 7-H_a GPTM, H-C_β Asp), 2.09 (d, J = 14.0 Hz, 1 H, 7-H_b GPTM), 1.85-1.77 (m, 1 H, 6-H_a GPTM), 1.73-1.42 (m, 5 H, 6-H_b GPTM, H-C_β Arg, H-C_γ Arg). ¹³C NMR (100 MHz, D₂O): δ = 179.8, 177.41, 177.36, 174.4, 173.7, 173.5 (s, CO), 159.3 (s, CN), 77.3 (s, C-7a GPTM), 58.9 (d, C-2 GPTM), 57.7 (d, C_α Arg), 51.6 (d, C_α Asp), 46.5 (t, C_α Gly), 43.5 (t, C-5 GPTM), 43.0 (t, C_δ Arg), 41.7 (t, C-1 GPTM), 37.1 (t, C_β Asp), 36.8 (t, C-7 GPTM), 28.7 (t, C_β Arg), 28.5 (t, C_γ Arg), 27.0 (t, C-6 GPTM). MS (ESI): 495 [MH⁺].

<A NAME="RG16706ST-16">16</A> Kumar CC. Nie H. Rogers CP. Malkowski M. Maxwell E. Catino JJ. Armstrong L. J. Pharm. Exp. Ther. 1997, 283: 843

The reproducibility of the IC₅₀ values was proved by performing the same tests on the antagonist cyclo(-Temp8-Arg-Gly-Asp-) reported by Belvisi et al. in ref. 6e.