Biochemical characterization of smoothened receptor antagonists by binding kinetics against drug-resistant mutant

Shimizu, Yūji; Ishii, Tsuyoshi; Ogawa, Kazumasa; Sasaki, Satoshi; Matsui, Hideki; Nakayama, Masaharu

doi:10.1016/j.ejphar.2015.05.062

Cited by 10 publications

(9 citation statements)

References 37 publications

(56 reference statements)

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“…Our data reject some previous observations on ear development using a less specific Smo antagonist, cyclopamine (Koebernick et al, ) but confirm observations on anterior translocation of hypaxial muscle fibers (Martin et al, ). Our data on ear size reduction and fusion of sensory epithelia are consistent with Shh function in proliferation regulation through n‐Myc (Yang et al, ; Kopecky et al, ) leading to tumor formation (Rudin, ; Morinello et al, ; Shimizu et al, ) and show that Shh affects the size of the ear.…”

Section: Discussionsupporting

confidence: 86%

“…Vismodegib (GDC‐0449) is an orally active Hedgehog pathway inhibitor with an IC 50 of 3 nM that also inhibits P‐gp and ABCG2 with IC 50 values of 3.0 and 1.4 μM, respectively. Vismodegib has been used in clinical trials and model organisms to antagonize the Shh/Smo induced hyperproliferation of tumors such as medulloblastoma (LoRusso et al, ; Morinello et al, ; Rudin, ; Shimizu et al, ). Given the conservation of Shh/Smo pathway in bilateria, this Smo antagonist will likely affect the Shh pathway in Xenopus , but details of drug interactions remain unknown.…”

Section: Introductionmentioning

confidence: 99%

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Sonic hedgehog antagonists reduce size and alter patterning of the frog inner ear

Zarei

Fritzsch

et al. 2017

Developmental Neurobiology

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Sonic Hedgehog (Shh) signaling plays a major role in vertebrate development, from regulation of proliferation to the patterning of various organs. In amniotes, Shh affects dorsoventral patterning in the inner ear but affects anteroposterior patterning in teleosts. Currently, it remains unknown the function of Shh in inner ear development in terms of how morphogenesis changes in the sarcopterygian/tetrapod lineage coincide with the evolution of limbs and novel auditory organs in the ear. In this study we used the tetrapod, Xenopus laevis, to test how increasing concentrations of the Shh signal pathway antagonist, Vismodegib, affects ear development. Vismodegib treatment dose dependently alters the development of the ear, hypaxial muscle, and indirectly the Mauthner cell through its interaction with the inner ear afferents. Together, these phenotypes have an effect on escape response. The altered Mauthner cell likely contributes to the increased time to respond to a stimulus. In addition, the increased hypaxial muscle in the trunk likely contributes to the subtle change in animal C-start flexion angle. In the ear, Vismodegib treatment results in decreasing segregation between the gravistatic sensory epithelia as the concentration of Vismodegib increases. Furthermore, at higher doses, there is a loss of the horizontal canal but no enantiomorphic transformation, as in bony fish lacking Shh. Like in amniotes, Shh signaling in frogs affects dorsoventral patterning in the ear, suggesting that auditory sensory evolution in sarcopterygians/tetrapods evolved with a shift of Shh axis specification of the ear.

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Section: Discussionsupporting

confidence: 86%

Section: Introductionmentioning

confidence: 99%

Sonic hedgehog antagonists reduce size and alter patterning of the frog inner ear

Zarei

Fritzsch

et al. 2017

Developmental Neurobiology

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“…One unfortunate side effect of SMO inhibition can be adaptive mutations in the SMO gene making them resistant to vismodegib or cyclopamine (170). An encouraging finding is that TAK-441 has been shown in preclinical studies to maintain activity in cells expressing these SMO adaptive mutants, suggesting it may be highly relevant to resistant patients (171,172). To date, there has only been clinical trial with TAK-441, a phase I trial that found it was well tolerated and showed preliminary antitumor activity in advanced solid tumors (173).…”

Section: Agents Targeting Smomentioning

confidence: 99%

Safety and Tolerability of Sonic Hedgehog Pathway Inhibitors in Cancer

Carpenter

Ray

2019

Drug Saf

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The hedgehog pathway, for which sonic hedgehog (Shh) is the most prominent ligand, is highly conserved and is tightly associated with embryonic development in a number of species. This pathway is also tightly associated with development of several types of cancer, including basal cell carcinoma and acute promyelocytic leukemia (APL) among many others. Inactivating mutations in Patched 1 (PTCH1), leading to ligand-independent pathway activation, are frequent in several cancer types but most prominent in basal cell carcinoma. This has led to the development of several compounds targeting this pathway as a cancer therapeutic. These compounds target the inducers of this pathway in Smoothened (SMO) and the GLI transcription factors, although targeting SMO has had the most success. Despite the many attempts at targeting this pathway, there are only three FDA-approved drugs for cancers that affect the Shh pathway. Two of these compounds, vismodegib and sonidegib, target SMO to suppress signaling from either PTCH1 or SMO mutations that lead to upregulation of the pathway. The other approved compound is arsenic trioxide (ATO), which can suppress this pathway at the level of the GLI proteins, although current evidence suggests it also has other targets. This review focuses on the efficacy and safety of these clinically-approved drugs targeting the Shh pathway along with a discussion on other Shh pathway inhibitors being developed.

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“…8 For simulation of actual data of [ 3 H]TAK-441 binding, k 1 and k 2 values were fixed to the reported values (1.5 × 10 6 M −1 min −1 and 9.7 × 10 −4 min −1 , respectively). 19…”

Section: Methodsmentioning

confidence: 99%

Characterization of Kinetic Binding Properties of Unlabeled Ligands via a Preincubation Endpoint Binding Approach

2016

Self Cite

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The dissociation rates of unlabeled drugs have been well studied by kinetic binding analyses. Since kinetic assays are laborious, we developed a simple method to determine the kinetic binding parameters of unlabeled competitors by a preincubation endpoint assay. The probe binding after preincubation of a competitor can be described by a single equation as a function of time. Simulations using the equation revealed the degree of IC50 change induced by preincubation of a competitor depended on the dissociation rate koff of the competitor but not on the association rate kon To validate the model, an in vitro binding assay was performed using a smoothened receptor (SMO) and [(3)H]TAK-441, a SMO antagonist. The equilibrium dissociation constants (KI) and koff of SMO antagonists determined by globally fitting the model to the concentration-response curves obtained with and without 24 h preincubation correlated well with those determined by other methods. This approach could be useful for early-stage optimization of drug candidates by enabling determination of binding kinetics in a high-throughput manner because it does not require kinetic measurements, an intermediate washout step during the reaction, or prior determination of competitors' KI values.

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Biochemical characterization of smoothened receptor antagonists by binding kinetics against drug-resistant mutant

Cited by 10 publications

References 37 publications

Sonic hedgehog antagonists reduce size and alter patterning of the frog inner ear

Sonic hedgehog antagonists reduce size and alter patterning of the frog inner ear

Safety and Tolerability of Sonic Hedgehog Pathway Inhibitors in Cancer

Characterization of Kinetic Binding Properties of Unlabeled Ligands via a Preincubation Endpoint Binding Approach

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