Cilliobrevin D, AAA+ ATPase Motor Cytoplasmic Dynein Inhibitor

Ciliobrevin D is the first specific small molecule antagonist of cytoplasmic dynein, and can inhibit both cytoplasmic dynein 1 and 2. The conversion of chemical energy into mechanical force by AAA+ ATPases is integral to cellular processes, including DNA replication, protein unfolding, cargo transport and membrane fusion. The AAA+ ATPase motor cytoplasmic dynein regulates ciliary trafficking, mitotic spindle formation and organelle transport. Ciliobrevin D perturbs protein trafficking within the primary cilium, leading to their malformation and Hedgehog signaling blockade. It also prevents spindle pole focusing, kinetochore–microtubule attachment, melanosome aggregation and peroxisomemotility in cultured cells. Ciliobrevin D can block dynein-dependent microtubule gliding and ATPase activity in vitro. Therefore it serves as a useful probe for dynein-dependent processes and studying cellular processes that require this microtubule motor.

Product information

CAS Number: 1370554-01-0

Molecular Weight: 392.62

Formula: C17H8Cl3N3O2

Chemical Name: 2-(7-chloro-4-oxo-3,4-dihydroquinazolin-2(1H)-ylidene)-3-(2,4-dichlorophenyl)-3-oxopropanenitrile

Smiles: N#CC(=C1NC(=O)C2=CC=C(Cl)C=C2N1)C(=O)C1=CC=C(Cl)C=C1Cl

InChiKey: JKKSCGHDRHRWOQ-VBKFSLOCSA-N

InChi: InChI=1S/C17H8Cl3N3O2/c18-8-1-3-10(13(20)5-8)15(24)12(7-21)16-22-14-6-9(19)2-4-11(14)17(25)23-16/h1-6,22H,(H,23,25)/b16-12-

Technical Data

Appearance: Solid Power.

Purity: ≥98% (or refer to the Certificate of Analysis)

Solubility: DMSO : 5 mg/mL (12.73 mM; ultrasonic and warming and heat to 80°C)

Shipping Condition: Shipped under ambient temperature as non-hazardous chemical or refer to Certificate of Analysis

Storage Condition: Dry, dark and -20 ^oC for 1 year or refer to the Certificate of Analysis.

Shelf Life: ≥360 days if stored properly.

Stock Solution Storage: 0 - 4 ^oC for 1 month or refer to the Certificate of Analysis.

Drug Formulation: To be determined.

HS Tariff Code: 382200

How to use

In Vitro:

Cells treated with Ciliobrevin D exhibits abnormal (unfocused, multipolar, or collapsed) spindles with disrupted γ-tubulin localization in NIH-3T3 cells. Similar Ciliobrevin-induced spindle defects are observed in HeLa cells, although to a lesser extent. Ciliobrevin D addition also reversibly disrupts the pre-formed spindles of metaphase-arrested cells and reduces overall microtubule levels. . Ciliobrevin D reversibly inhibits melanosome aggregation, but the non-cilia-disrupting derivative had no discernible effect at comparable doses. Ciliobrevin D similarly abrogates the movement of peroxisomes in Drosophila S2 cells.

In Vivo:

Knockdown of Dync1h1 or inactivation of dynein 1 by Ciliobrevin D in the testis in vivo perturbs spermatogenesis. Knockdown of Dync1h1 or the use of Ciliobrevin D to inactivate dynein 1 in the testis in vivo perturbs MT organization through changes in the spatial expression of EB1, perturbs F-actin organization, and perturbs distribution of adhesion protein complexes at the BTB, leading to a loss of BTB integrity. F-actin disorganization in the seminiferous epithelium following Dync1h1 knockdown or dynein 1 inactivation by Ciliobrevin D is mediated by changes in the spatiotemporal expression of actin regulatory proteins Arp3 and Eps8.

References:

1. Firestone AJ, et al. Small-molecule inhibitors of the AAA+ ATPase motor cytoplasmic dynein. (2012) Nature. 484(7392):125-9.
2. Miao Y, et al. Dynein promotes porcine oocyte meiotic progression by maintaining cytoskeletal structures and cortical granule arrangement. Cell Cycle. 2017;16(21):2139-2145.
3. Wen Q, et al. Dynein 1 supports spermatid transport and spermiation during spermatogenesis in the rat testis. Am J Physiol Endocrinol Metab. 2018 Nov 1;315(5):E924-E948.

Products are for research use only. Not for human use.