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. 2018 Jan 29;9(3):227-232.
doi: 10.1021/acsmedchemlett.7b00500. eCollection 2018 Mar 8.

In Search of the Optimal Macrocyclization Site for Neurotensin

Marc Sousbie  1 , Élie Besserer-Offroy  1 , Rebecca L Brouillette  1 , Jean-Michel Longpré  1 , Richard Leduc  1 , Philippe Sarret  1 , Éric Marsault  1
Affiliations

In Search of the Optimal Macrocyclization Site for Neurotensin

Marc Sousbie et al. ACS Med Chem Lett. .

Abstract

Neurotensin exerts potent analgesic effects following activation of its cognate GPCRs. In this study, we describe a systematic exploration, using structure-based design, of conformationally constraining neurotensin (8-13) with the help of macrocyclization and the resulting impacts on binding affinity, signaling, and proteolytic stability. This exploratory study led to a macrocyclic scaffold with submicromolar binding affinity, agonist activity, and greatly improved plasma stability.

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Conflict of interest statement

The authors declare no competing financial interest.

Figures

Scheme 1
Scheme 1. Macrocyclization Reaction
Reagents and conditions: (a) Hoveyda–Grubbs second generation catalyst, benzoquinone, DCE, 50 °C, microwave, 1 h; (b) TFA/DCM/TIS, 1 h.
Figure 1
Figure 1
Structures of neurotensin (8–13) and macrocyclic analogues explored in this study.
Figure 2
Figure 2
Custom linkers A and B.
Figure 3
Figure 3
Top row: Macrocycles 1, 2, and 6 (A, B, and C, respectively) aligned with the conformation of NT (8–13) within the crystal structure of NTS1 (green). Bottom row: Macrocycles 7, 8, and 9 (D, E, and F, respectively) docked into NTS1. NT (8–13) (green) is shown for comparison. Molecular graphics were performed with the UCSF Chimera package.
Figure 4
Figure 4
Comparison of the crystallized NT (8–13) conformation with 10. (A) NTS1 crystal. Blue arrow indicates available space between Tyr11 and Arg8. (B) Superimposition of NT (8–13) from the crystal and macrocycle 10 (docked). (C) Macrocycle 10 docked into a homology model of human NTS1 based on the crystal. Molecular graphics were performed with the UCSF Chimera package.
Figure 5
Figure 5
Plasma stability of 10 compared to NT (8–13).
See this image and copyright information in PMC

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