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

Name: Spebrutinib
Brand: MedChemExpress
SKU: HY-18012
Rating: 5.0 (13 reviews)

Cat. No.: HY-18012 Purity: 99.62%: FAQs
Data Sheet SDS COA Handling Instructions

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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 Chemical Structure

CAS No. : 1202757-89-8

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Solid + Solvent (Highly Recommended)
10 mM * 1 mL in DMSO ready for reconstitution	USD 73	In-stock	Estimated Time of Arrival: December 31
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10 mM * 1 mL in DMSO	USD 73	In-stock	Estimated Time of Arrival: December 31
Solid
5 mg	USD 66	In-stock	Estimated Time of Arrival: December 31
10 mg	USD 99	In-stock	Estimated Time of Arrival: December 31
50 mg	USD 165	In-stock	Estimated Time of Arrival: December 31
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Customer Review

Based on 13 publication(s) in Google Scholar

Other Forms of Spebrutinib:

Spebrutinib besylate Get quote

12 Publications Citing Use of MCE Spebrutinib

: Spebrutinib purchased from MedChemExpress. Usage Cited in: Blood. 2016 Jun 23;127(25):3237-52. [Abstract]; Ibrutinib binds to the ATP-binding pocketof HCK and blocks ATP binding. Results from kinase active-site inhibition assaysutilizing an ATP-BTN probe that is used to pull downactive kinases in the presence of Ibrutinib, CC-292, or A419259 in lysates from BCWM.1 WM cells.

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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]
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]
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]
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]
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]
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]
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]
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 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]
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]
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.

Clinical Trial

Molecular Weight

423.44

Formula

C₂₂H₂₂FN₅O₃

CAS No.

1202757-89-8

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
10 mM	0.2362 mL	1.1808 mL	2.3616 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)

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

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

Concentration _(start)

Volume _(start)

Concentration _(final)

Volume _(final)

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:

Dosage

mg/kg

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(per animal)

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(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:

DMSO +

Tween-80 +

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: 99.62%

Select Batch:

Data Sheet (273 KB) SDS (393 KB)

COA (246 KB) LCMS (265 KB)

Handling Instructions (2659 KB)

References

[1]. Evans EK, et al. Inhibition of Btk with CC-292 provides early pharmacodynamic assessment of activity in mice and humans. J Pharmacol Exp Ther. 2013 Aug;346(2):219-28. [Content Brief]

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

[1]. Evans EK, et al. Inhibition of Btk with CC-292 provides early pharmacodynamic assessment of activity in mice and humans. J Pharmacol Exp Ther. 2013 Aug;346(2):219-28. [Content Brief]

Complete Stock Solution Preparation Table

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

Spebrutinib Related Classifications

Help & FAQs

Do most proteins show cross-species activity?
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

Spebrutinib1202757-89-8AVL-292 CC-292AVL292AVL 292CC292CC 292CC-292BtkBruton tyrosine kinaseInhibitorinhibitorinhibit

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

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Other Forms of Spebrutinib:

Spebrutinib Related Antibodies

12 Publications Citing Use of MCE Spebrutinib

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Customer Review

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Complete Stock Solution Preparation Table

Spebrutinib Related Classifications

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