1. Protein Tyrosine Kinase/RTK
  2. Btk
  3. Spebrutinib

Spebrutinib  (Synonyms: AVL-292; CC-292)

Cat. No.: HY-18012 Purity: 99.22%
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Spebrutinib (AVL-292; CC-292) is a covalent, orally active, and highly selective with an IC50 of 0.5 nM.

For research use only. We do not sell to patients.

Spebrutinib

Spebrutinib Chemical Structure

CAS No. : 1202757-89-8

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Customer Review

Based on 17 publication(s) in Google Scholar

Other Forms of Spebrutinib:

Top Publications Citing Use of Products
  • Biological Activity

  • Protocol

  • Purity & Documentation

  • References

  • Customer Review

Description

Spebrutinib (AVL-292; CC-292) is a covalent, orally active, and highly selective with an IC50 of 0.5 nM.

IC50 & Target

IC50: <0.5 nM (Btk)[1]

Cellular Effect
Cell Line Type Value Description References
786-0 IC50
14.9 μM
Compound: 3
Cytotoxicity against human 786-O cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human 786-O cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
A549 IC50
1.98 μM
Compound: 3
Cytotoxicity against human A549 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human A549 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
B-cell IC50
3.28 nM
Compound: 2
Antiproliferative activity against B cells (unknown origin)
Antiproliferative activity against B cells (unknown origin)
[PMID: 26976214]
BJ IC50
27.6 μM
Compound: 3
Cytotoxicity against human BJ cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human BJ cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
HEF IC50
22.6 μM
Compound: 3
Cytotoxicity against human HEF cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human HEF cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
HEL IC50
5.3 μM
Compound: CC292
Antiproliferative activity against human HEL cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
Antiproliferative activity against human HEL cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
[PMID: 31843459]
HT-29 IC50
18.9 μM
Compound: 3
Cytotoxicity against human HT-29 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human HT-29 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
HeLa IC50
17.4 μM
Compound: 3
Cytotoxicity against human HeLa cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human HeLa cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
HepG2 IC50
33.5 μM
Compound: 3
Cytotoxicity against human HepG2 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human HepG2 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
JeKo-1 IC50
1.3 μM
Compound: CC292
Antiproliferative activity against human JeKo1 cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
Antiproliferative activity against human JeKo1 cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
[PMID: 31843459]
JeKo-1 IC50
1480 nM
Compound: 8; Spe
Antiproliferative activity against human JeKo-1 cells assessed as inhibition of cell growth incubated for 72 hrs by CCK8 assay
Antiproliferative activity against human JeKo-1 cells assessed as inhibition of cell growth incubated for 72 hrs by CCK8 assay
[PMID: 36399923]
Jurkat IC50
6.7 μM
Compound: 3
Cytotoxicity against human Jurkat cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human Jurkat cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
K562 IC50
12.6 μM
Compound: 3
Cytotoxicity against human K562 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human K562 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
K562 IC50
7400 nM
Compound: 8; Spe
Antiproliferative activity against human K562 cells assessed as inhibition of cell growth incubated for 72 hrs by CCK8 assay
Antiproliferative activity against human K562 cells assessed as inhibition of cell growth incubated for 72 hrs by CCK8 assay
[PMID: 36399923]
KARPAS-299 IC50
20.3 μM
Compound: 3
Cytotoxicity against human KARPAS299 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human KARPAS299 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
MCF-10A IC50
4.36 μM
Compound: 3
Cytotoxicity against human MCF10A cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human MCF10A cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
MCF7 IC50
29 μM
Compound: 3
Cytotoxicity against human MCF7 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human MCF7 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
MDA-MB-231 IC50
> 40 μM
Compound: 3
Cytotoxicity against human MDA-MB-231 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human MDA-MB-231 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
MOLT-4 IC50
1463 nM
Compound: 8; Spe
Antiproliferative activity against human MOLT-4 cells assessed as inhibition of cell growth incubated for 72 hrs by CCK8 assay
Antiproliferative activity against human MOLT-4 cells assessed as inhibition of cell growth incubated for 72 hrs by CCK8 assay
[PMID: 36399923]
MV4-11 IC50
245 nM
Compound: 8; Spe
Antiproliferative activity against human MV4-11 cells assessed as inhibition of cell growth incubated for 72 hrs by CCK8 assay
Antiproliferative activity against human MV4-11 cells assessed as inhibition of cell growth incubated for 72 hrs by CCK8 assay
[PMID: 36399923]
NAMALVA IC50
3.42 μM
Compound: Spebrutinib
Antiproliferative activity against human NAMALWA cells after 72 hrs by CCK-8 assay
Antiproliferative activity against human NAMALWA cells after 72 hrs by CCK-8 assay
[PMID: 29146136]
NCI-H1299 IC50
38.5 μM
Compound: 3
Cytotoxicity against human H1299 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human H1299 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
NCI-H3122 IC50
32.5 μM
Compound: 3
Cytotoxicity against human NCI-H3122 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human NCI-H3122 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
OCI-Ly10 IC50
> 10 μM
Compound: CC292
Antiproliferative activity against human OCI-LY10 cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
Antiproliferative activity against human OCI-LY10 cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
[PMID: 31843459]
PC-3 IC50
22.2 μM
Compound: 3
Cytotoxicity against human PC3 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
Cytotoxicity against human PC3 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
[PMID: 31395509]
Raji IC50
17.6 μM
Compound: CC292
Antiproliferative activity against human Raji cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
Antiproliferative activity against human Raji cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
[PMID: 31843459]
Raji IC50
21.6 μM
Compound: 3
Antiproliferative activity against human Raji cells assessed as reduction in cell growth incubated for 48 hrs by MTS assay
Antiproliferative activity against human Raji cells assessed as reduction in cell growth incubated for 48 hrs by MTS assay
[PMID: 31395509]
Raji IC50
25.3 μM
Compound: 4; AVL-292
Antiproliferative activity against human Raji cells after 48 hrs by CCK-8 assay
Antiproliferative activity against human Raji cells after 48 hrs by CCK-8 assay
[PMID: 27994736]
Raji IC50
25.3 μM
Compound: Spebrutinib
Antiproliferative activity against human Raji cells measured after 48 hrs by CCK-8 assay
Antiproliferative activity against human Raji cells measured after 48 hrs by CCK-8 assay
[PMID: 27956037]
Raji IC50
25.3 μM
Compound: Spebrutinib
Antiproliferative activity against human Raji cells after 72 hrs by CCK-8 assay
Antiproliferative activity against human Raji cells after 72 hrs by CCK-8 assay
[PMID: 29146136]
Raji IC50
29.4 μM
Compound: 4
Antiproliferative activity against human Raji cells after 48 hrs by CCK-8 assay
Antiproliferative activity against human Raji cells after 48 hrs by CCK-8 assay
[PMID: 27912175]
Raji IC50
29.4 μM
Compound: 4
Antiproliferative activity against human Raji cells over expressing BTK after 48 hrs by CCK-8 assay
Antiproliferative activity against human Raji cells over expressing BTK after 48 hrs by CCK-8 assay
[PMID: 28432946]
Ramos IC50
14.2 μM
Compound: Spebrutinib
Antiproliferative activity against human Ramos cells after 72 hrs by CCK-8 assay
Antiproliferative activity against human Ramos cells after 72 hrs by CCK-8 assay
[PMID: 29146136]
Ramos IC50
15.1 μM
Compound: 4; AVL-292
Antiproliferative activity against human Ramos cells after 48 hrs by CCK-8 assay
Antiproliferative activity against human Ramos cells after 48 hrs by CCK-8 assay
[PMID: 27994736]
Ramos IC50
15.1 μM
Compound: Spebrutinib
Antiproliferative activity against human Ramos cells measured after 48 hrs by CCK-8 assay
Antiproliferative activity against human Ramos cells measured after 48 hrs by CCK-8 assay
[PMID: 27956037]
Ramos IC50
18.8 μM
Compound: 4
Antiproliferative activity against human Ramos cells over expressing BTK after 48 hrs by CCK-8 assay
Antiproliferative activity against human Ramos cells over expressing BTK after 48 hrs by CCK-8 assay
[PMID: 28432946]
Ramos IC50
20.9 μM
Compound: 4
Antiproliferative activity against human Ramos cells after 48 hrs by CCK-8 assay
Antiproliferative activity against human Ramos cells after 48 hrs by CCK-8 assay
[PMID: 27912175]
Ramos IC50
3.9 μM
Compound: CC292
Antiproliferative activity against human Ramos cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
Antiproliferative activity against human Ramos cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
[PMID: 31843459]
Ramos IC50
5.44 μM
Compound: 3
Antiproliferative activity against human Ramos cells assessed as reduction in cell growth incubated for 48 hrs by MTS assay
Antiproliferative activity against human Ramos cells assessed as reduction in cell growth incubated for 48 hrs by MTS assay
[PMID: 31395509]
Sf9 IC50
0.6 nM
Compound: 4; AVL-292
Inhibition of recombinant human N-terminal His-tagged BTK expressed in baculovirus infected sf9 cells using poly(4:1 Glu,Tyr) as substrate by ADP-Glo kinase assay
Inhibition of recombinant human N-terminal His-tagged BTK expressed in baculovirus infected sf9 cells using poly(4:1 Glu,Tyr) as substrate by ADP-Glo kinase assay
[PMID: 27994736]
Sf9 IC50
0.72 nM
Compound: Spebrutinib
Inhibition of human recombinant full-length N-terminal His-tagged BTK expressed in baculovirus infected Sf9 insect cells measured after 60 mins by ADP-Glo kinase assay
Inhibition of human recombinant full-length N-terminal His-tagged BTK expressed in baculovirus infected Sf9 insect cells measured after 60 mins by ADP-Glo kinase assay
[PMID: 27956037]
Sf9 IC50
2.12 nM
Compound: 3
Inhibition of human recombinant full length BTK expressed in baculovirus in Sf9 insect cells using Poly (4:1 Glu, Tyr) as substrate incubated for 1 hr by ADP-Glo assay
Inhibition of human recombinant full length BTK expressed in baculovirus in Sf9 insect cells using Poly (4:1 Glu, Tyr) as substrate incubated for 1 hr by ADP-Glo assay
[PMID: 31395509]
Sf9 IC50
66.5 nM
Compound: Spebrutinib
Inhibition of recombinant human N-terminal GST-tagged JAK3 (781 to end residues) expressed in baculovirus infected Sf9 cells using poly (Glu,Tyr) 4:1 as substrate incubated for 60 mins in presence of ATP by ADP-Glo kinase assay
Inhibition of recombinant human N-terminal GST-tagged JAK3 (781 to end residues) expressed in baculovirus infected Sf9 cells using poly (Glu,Tyr) 4:1 as substrate incubated for 60 mins in presence of ATP by ADP-Glo kinase assay
[PMID: 31866272]
In Vitro

Spebrutinib (CC-292) is a covalent, highly selective, orally active inhibitor of Btk with IC50 value of 0.5 nM. Spebrutinib also less potently inhibits Yes, c-Src, Brk, Lyn, and Fyn with IC50s of 723 nM, 1.729 μM, 2.43 μM, 4.4 μM, and 7.15 μM, rspectively. Extensive analysis has revealed that the EC50 of Btk occupancy from a Spebrutinib dose-response in Ramos cells (EC50=6 nM) correlated directly with the cellular EC50 of Btk kinase inhibition with Spebrutinib (EC50=8 nM). Furthermore, the concentration at which Spebrutinib inhibits 90% of Btk activity in Ramos cells is 35 nM while the concentration of Spebrutinib required for 90% occupancy of Btk is 39 nM[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Molecular Weight

423.44

Formula

C22H22FN5O3

CAS No.
Appearance

Solid

Color

White to khaki

SMILES

COCCOC1=CC=C(NC2=NC=C(F)C(NC3=CC=CC(NC(C=C)=O)=C3)=N2)C=C1

Shipping

Room temperature in continental US; may vary elsewhere.

Storage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 2 years
-20°C 1 year
Solvent & Solubility
In Vitro: 

DMSO : ≥ 45 mg/mL (106.27 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

*"≥" means soluble, but saturation unknown.

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.3616 mL 11.8080 mL 23.6161 mL
5 mM 0.4723 mL 2.3616 mL 4.7232 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
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Volume
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Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

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V1

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C2

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In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.5 mg/mL (5.90 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% Corn Oil

    Solubility: ≥ 2.5 mg/mL (5.90 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.

In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

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mg/kg

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(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
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+
%
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%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation

Purity: 99.22%

References
Cell Assay
[1]

Cells are incubated in serum-free RPMI media for 1-1.5 hours. Isolated human B cells are incubated with Spebrutinib at a final concentration of 0.001, 0.01, 0.1 and 1 μM. Ramos cells are incubated with 0.1 nM-3 μM Spebrutinib. Cells are then incubated in the presence of compound for 1 hour at 37°C. Following incubation, cells are centrifuged and resuspended in 100 μL of serum-free RPMI and BCR is stimulated with addition of 5 μg/mL α-human IgM. Samples are centrifuged, washed in phosphate-buffered saline (PBS), and lysed in 100 μL of Cell Extraction Buffer plus 1:10 (v/v) PhosSTOP Phosphatase Inhibitor and 1:10 (v/v) Complete Protease Inhibitor. Antibodies used for immunoblot analysis include P-PLCγ2, PLCγ2 (3871; CST), Syk (2712; CST), P-Syk (2710; CST), Btk, P-Btk, and Tubulin. Membranes are scanned on a Li-Cor Odyssey scanner using infrared fluorescence detection[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 2.3616 mL 11.8080 mL 23.6161 mL 59.0402 mL
5 mM 0.4723 mL 2.3616 mL 4.7232 mL 11.8080 mL
10 mM 0.2362 mL 1.1808 mL 2.3616 mL 5.9040 mL
15 mM 0.1574 mL 0.7872 mL 1.5744 mL 3.9360 mL
20 mM 0.1181 mL 0.5904 mL 1.1808 mL 2.9520 mL
25 mM 0.0945 mL 0.4723 mL 0.9446 mL 2.3616 mL
30 mM 0.0787 mL 0.3936 mL 0.7872 mL 1.9680 mL
40 mM 0.0590 mL 0.2952 mL 0.5904 mL 1.4760 mL
50 mM 0.0472 mL 0.2362 mL 0.4723 mL 1.1808 mL
60 mM 0.0394 mL 0.1968 mL 0.3936 mL 0.9840 mL
80 mM 0.0295 mL 0.1476 mL 0.2952 mL 0.7380 mL
100 mM 0.0236 mL 0.1181 mL 0.2362 mL 0.5904 mL
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Spebrutinib
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