Siponimod (BAF312)

Name: Siponimod (BAF312)
Brand: Selleck Chemicals
SKU: S7179
Rating: 5 (5 reviews)

For research use only.

Catalog No.S7179

5 publications

CAS No. 1230487-00-9

BAF312 (Siponimod) is a next-generation S1P receptor agonist, selective for S1P1 and S1P5 receptors with EC50 of 0.39 nM and 0.98 nM, exhibits >1000-fold selectivity over S1P2, S1P3 and S1P4 receptors. Phase 3.

Selleck's Siponimod (BAF312) has been cited by 5 publications

Front Immunol, 2020, 8;11:635

PubMed: 32322257 ( click the link to review the publication )

Int J Nanomedicine, 2020, 15:5561-5571

PubMed: 32801704 ( click the link to review the publication )

Leukemia, 2019, 10.1038/s41375-019-0478-9

PubMed: 31097785 ( click the link to review the publication )

Leukemia, 2019, 10.1038/s41375-019-0511-z

PubMed: 28878352 ( click the link to review the publication )

Int J Mol Sci, 2018, 19(5)

PubMed: 29772789 ( click the link to review the publication )

1 Customer Review

Immunoblot of KMH2 cells following 1 h pre-treatment with increasing concentrations of Ozanimod or Siponimod.

Leukemia, 2018, 32(1):214-223. Siponimod (BAF312) purchased from Selleck.

Purity & Quality Control

Choose Selective S1P Receptor Inhibitors

Biological Activity

Description

Targets

S1P1 receptor ^[1]	S1P5 receptor ^[1]
0.39 nM(EC50)	0.98 nM(EC50)

In vitro

BAF312 (Siponimod) is a potent and selective S1P receptor agonist, with EC50 of 0.39 nM and 0.98 nM for S1P₁ and S1P₅receptors, exhibits >1000-fold selectivity over S1P₂, S1P₃ and S1P₄ receptors. ^[1]BAF312 (1 h at 1 μM) promotes prominent internalization of S1P1 receptors by 91%.^[2]

Assay

Methods	Test Index	PMID
Western blot	p-ERK / ERK / p-AKT / AKT ; PubMed: 26856814 J Neuroinflammation Agonism of S1PR1/5 promotes pERK and pAkt signalling in mouse and human astrocytes. BAF312 at concentrations shown induced pERK after 10 min (n = 3) and pAKT after 30 min treatments (n = 4) a in mouse astrocytes and b in human astrocytes. Data presented as ±SEM (n = 3-4), one-way ANOVA and Newman-Keuls multiple comparison post-test. p < 0.05, p < 0.01, **p < 0.001.	26856814
Immunofluorescence	Vimentin / S1PR1 ; PubMed: 26856814 J Neuroinflammation BAF312 induces internalisation of the S1PR1. Mouse astrocytes were serum starved for 4 h and then treated for 1 h with BAF312 (10 nM, 100 nM, 1 μM) or pFTY (1 μM). BAF312 at 1 μM concentration induced internalisation of S1PR1.	26856814

In vivo

BAF312 effectively suppresses encephalomyelitis (EAE) in rats by internalizing S1P₁ receptors, rendering them insensitive to the egress signal from lymph nodes. ^[1] BAF312 significantly reduces clinical scores when dosed prophylactically or therapeutically in mice at 0.3 mg/kg. ^[3]

Protocol

Kinase Assay:^[1]	- Collapse GTPγ[³⁵S] binding assay: The cells are homogenized and centrifuged at 26900 × g for 30 min at 4°C. Membranes are re-suspended in 20 mM HEPES (pH 7.4), 100 mM NaCl, 10 mM MgCl₂, 1 mM EDTA and 0.1% fat-free BSA at 2–3 mg protein/mL. GTPγ[³⁵S] binding assay is performed with the membranes (75 mg protein /mL in 50 mM HEPES, 100 mM NaCl, 10 mM MgCl₂, 20 μg/mL saponin and 0.1% fat-free BSA (pH 7.4), 5 mg/mL with wheat-germ agglutinin-coated scintillation proximity assay-bead, and 10 μM GDP for 10–15 min. The GTPγ[³⁵S]-binding reaction is started by the addition of 200 pM GTPγ[³⁵S]. After 120 min at room temperature, the plates are centrifuged for 10 min at 300 × g and counted.
Cell Research: ^[1]	- Collapse Cell lines: CHO Concentrations: ~1 μM Incubation Time: 1 h Method: Agonist-mediated internalization of S1P₁ receptors in CHO cells analysed by flow cytometry Myc-tag hS1P₁ cells are incubated for 1 h with agonist at 37°C in standard culture medium followed by a PBS wash. An aliquot is kept on ice for 3 h, while another aliquot is left for 3 (or 12) h in culture medium (no agonist) at 37°C. The cells are then incubated either with 4 μg/mL monoclonal mouse anti C-myc IgG1 antibody or with isotype control mouse IgG1 for 60 min at 4°C, followed by an incubation with 1 μg/mL of Alexa488-labelled goat anti-mouse secondary conjugates. The cells are subjected to flow cytometry measurements using 10000 viable cells per sample. (Only for Reference)
Animal Research:^[1]	- Collapse Animal Models: encephalomyelitis (EAE) model rat Dosages: 0.03, 0.3 and 3 mg/kg Administration: oral gavage (Only for Reference)

References

Solubility (25°C)

In vitro	DMSO	100 mg/mL warmed (193.57 mM)
	Water	Insoluble
	Ethanol	''44 mg/mL warmed

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information Download Siponimod (BAF312) SDF

Molecular Weight	516.6
Formula	C₂₉H₃₅F₃N₂O₃
CAS No.	1230487-00-9
Storage	3 years-20°C powder 2 years-80°C in solvent
Synonyms	N/A
Smiles	CCC1=C(C=CC(=C1)C(=NOCC2=CC(=C(C=C2)C3CCCCC3)C(F)(F)F)C)CN4CC(C4)C(=O)O

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

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Step 2: Enter the in vivo formulation (Different batches have different solubility ratios, please contact Selleck to provide you with the correct ratio)

% DMSO+ % + % Tween 80+ % ddH₂O

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Calculation results:

Working concentration： mg/ml；

Method for preparing DMSO master liquid: ： mg drug pre-dissolved in μL DMSO (Master liquid concentration mg/mL， Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation：Take DMSO master liquid, next addμL PEG300， mix and clarify, next addμL Tween 80，mix and clarify, next add μL ddH₂O，mix and clarify.

Note：1.Please make sure the liquid is clear before adding the next solvent.
2.Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.

Bio Calculators

Molarity Calculator

Calculate the mass, volume or concentration required for a solution. The Selleck molarity calculator is based on the following equation:

Mass (mg) = Concentration (mM) × Volume (mL) × Molecular Weight (g/mol)

*When preparing stock solutions, please always use the batch-specific molecular weight of the product found on the via label and MSDS / COA (available on product pages).

Dilution Calculator

Calculate the dilution required to prepare a stock solution. The Selleck dilution calculator is based on the following equation:

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

This equation is commonly abbreviated as: C1V1 = C2V2 ( Input Output )

* When preparing stock solutions always use the batch-specific molecular weight of the product found on the vial label and MSDS / COA (available online).

The Serial Dilution Calculator Equation

Serial Dilutions
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Computed Result

C1=C0/X	C1:	LOG(C1):
C2=C1/X	C2:	LOG(C2):
C3=C2/X	C3:	LOG(C3):
C4=C3/X	C4:	LOG(C4):
C5=C4/X	C5:	LOG(C5):
C6=C5/X	C6:	LOG(C6):
C7=C6/X	C7:	LOG(C7):
C8=C7/X	C8:	LOG(C8):

Molecular Weight Calculator

Enter the chemical formula of a compound to calculate its molar mass and elemental composition:

Tip: Chemical formula is case sensitive. C10H16N2O2 c10h16n2o2

Instructions to calculate molar mass (molecular weight) of a chemical compound:

To calculate molar mass of a chemical compound, please enter its chemical formula and click 'Calculate'.

Definitions of molecular mass, molecular weight, molar mass and molar weight:

Molecular mass (molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.

Molarity Calculator

Clinical Trial Information (data from https://clinicaltrials.gov, updated on 2020-09-01)

NCT Number	Recruitment	interventions	Conditions	Sponsor/Collaborators	Start Date	Phases
NCT03623243	Recruiting	Drug: Siponimod	Multiple Sclerosis Relapsing MS Advancing Multiple Sclerosis	Novartis Pharmaceuticals\|Novartis	February 14 2019	Phase 3
NCT03338998	Suspended	Drug: BAF312\|Drug: Placebo	Hemorrhagic Stroke\|Intracerebral Hemorrhage (ICH)	Novartis Pharmaceuticals\|Novartis	December 24 2017	Phase 2
NCT02029274	Terminated	Drug: BAF312\|Drug: Placebo	Active Dermatomyositis	Novartis Pharmaceuticals\|Novartis	August 25 2013	Phase 2
NCT01904214	Completed	Drug: BAF312	Renal Impairment	Novartis Pharmaceuticals\|Novartis	July 2013	Phase 1
NCT01801917	Terminated	Drug: Placebo\|Drug: BAF312	Polymyositis	Novartis Pharmaceuticals\|Novartis	April 24 2013	Phase 2
NCT01665144	Active not recruiting	Drug: BAF312\|Drug: Placebo	Secondary Progressive Multiple Sclerosis	Novartis Pharmaceuticals\|Novartis	December 20 2012	Phase 3

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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Siponimod (BAF312)