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5J-4

Alias: 5J 4 5J4 5J-4
Cat No.:V9662 Purity: ≥98%
5J-4 is a potent CRAC inhibitor.
5J-4
5J-4 Chemical Structure CAS No.: 827001-82-1
Product category: New1
This product is for research use only, not for human use. We do not sell to patients.
Size Price Stock Qty
1mg
5mg
10mg
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Product Description
5J-4 is a potent CRAC inhibitor. 5J-4 reduced the number of monocytes infiltrating into the CNS and significantly reduced infiltrating CD4+ cells. 5J-4 reduces symptoms and delays the onset of EAE (experimental autoimmune encephalomyelitis) in a mouse model of inflammation.
Biological Activity I Assay Protocols (From Reference)
ln Vivo
5J-4 decreases the expression of RORα and RORγt and the generation of IL-17 [1]. The intraperitoneal administration of 5J-4 (2 mg/kg) every other day for 30 days alleviates symptoms and postpones the start of EAE [1].
Animal Protocol
Animal/Disease Models: C57BL/6 mice (MOG35-55 peptide immunized mice) [1]
Doses: 2 m/kg
Route of Administration: intraperitoneal (ip) injection, every other day, for 30 days
Experimental Results: Dramatically diminished symptoms and delayed EAE attacks and reduce the incidence of EAE. The number of monocytes infiltrating into the central nervous system and Dramatically reducing the number of infiltrating CD4+ cell populations.
References

[1]. Calcium signaling via Orai1 is essential for induction of the nuclear orphan receptor pathway to drive Th17 differentiation. J Immunol. 2014 Jan 1;192(1):110-22.

These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C16N2O3SH12
Molecular Weight
312.3431
Exact Mass
312.056
CAS #
827001-82-1
PubChem CID
2968289
Appearance
White to off-white solid powder
LogP
3.8
Hydrogen Bond Donor Count
3
Hydrogen Bond Acceptor Count
4
Rotatable Bond Count
2
Heavy Atom Count
22
Complexity
432
Defined Atom Stereocenter Count
0
SMILES
OC1C=C2C(=CC=1)C(NC(=S)NC(C1OC=CC=1)=O)=CC=C2
InChi Key
WMUSJLJASFXGHN-UHFFFAOYSA-N
InChi Code
InChI=1S/C16H12N2O3S/c19-11-6-7-12-10(9-11)3-1-4-13(12)17-16(22)18-15(20)14-5-2-8-21-14/h1-9,19H,(H2,17,18,20,22)
Chemical Name
N-((6-hydroxynaphthalen-1-yl)carbamothioyl)furan-2-carboxamide
Synonyms
5J 4 5J4 5J-4
HS Tariff Code
2934.99.9001
Storage

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Shipping Condition
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
Solubility Data
Solubility (In Vitro)
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
Solubility (In Vivo)
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

Injection Formulations
(e.g. IP/IV/IM/SC)
Injection Formulation 1: DMSO : Tween 80: Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution 50 μL Tween 80 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO 400 μLPEG300 50 μL Tween 80 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).
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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C,1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO 100 μLPEG300 200 μL castor oil 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10: 10 : 80 (i.e. 100 μL Ethanol 100 μL Cremophor 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH 400 μLPEG300 50 μL Tween 80 450 μL Saline)


Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).
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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders


Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 3.2016 mL 16.0082 mL 32.0164 mL
5 mM 0.6403 mL 3.2016 mL 6.4033 mL
10 mM 0.3202 mL 1.6008 mL 3.2016 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.

Calculator

Molarity Calculator allows you to calculate the mass, volume, and/or concentration required for a solution, as detailed below:

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An example of molarity calculation using the molarity calculator is shown below:
What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
  • Enter 350.26 in the Molecular Weight (MW) box
  • Enter 10 in the Concentration box and choose the correct unit (mM)
  • Enter 5 in the Volume box and choose the correct unit (mL)
  • Click the “Calculate” button
  • The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

Dilution Calculator allows you to calculate how to dilute a stock solution of known concentrations. For example, you may Enter C1, C2 & V2 to calculate V1, as detailed below:

What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
  • Enter 10 into the Concentration (Start) box and choose the correct unit (mM)
  • Enter 25 into the Concentration (End) box and select the correct unit (mM)
  • Enter 25 into the Volume (End) box and choose the correct unit (mL)
  • Click the “Calculate” button
  • The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box
g/mol

Molecular Weight Calculator allows you to calculate the molar mass and elemental composition of a compound, as detailed below:

Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
Instructions to calculate molar mass (molecular weight) of a chemical compound:
  • To calculate molar mass of a chemical compound, please enter the chemical/molecular formula and click the “Calculate’ button.
Definitions of molecular mass, molecular weight, molar mass and molar weight:
  • Molecular mass (or 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.
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Reconstitution Calculator allows you to calculate the volume of solvent required to reconstitute your vial.

  • Enter the mass of the reagent and the desired reconstitution concentration as well as the correct units
  • Click the “Calculate” button
  • The answer appears in the Volume (to add to vial) box
In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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Calculation results

Working concentration mg/mL;

Method for preparing DMSO stock solution mg drug pre-dissolved in μL DMSO (stock solution 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 μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.

(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
             (2) Be sure to add the solvent(s) in order.

Biological Data
  • Compound 5D and 5J-4 ameliorate TH17-mediated autoimmune disease (A) EAE disease course in C57BL/6 mice injected intraperitoneally with either carrier alone or compound 5D (1 mg/kg) every alternate day starting from day −1. At day 0, disease was induced with injection of MOG35-55/CFA. The graph shows average ± s.e.m. and is representative of three experiments with 8-10 animals in each experiment. (B) Chemical structures of compound 5D and its analogue, compound 5J-4. (C) Measurement of block of SOCE by compound 5J-4 in HeLa-OSN cells (left panel) or primary TH17 cells (right panel). In all the graphs data represent average ± s.e.m. from 25-35 different cells. (D) Compound 5J-4 ameliorates symptoms of experimental autoimmune encephalomyelitis (EAE) in vivo. EAE disease course in mice injected intraperitoneally with either DMSO or compound 5J-4 (2 mg/kg) every alternate day starting from day 0 after disease induction with MOG35-55/CFA. The graph shows average ± s.e.m. from one of three independent repeats of the experiments with 10-20 mice per trial. (E) Compound 5J-4 reduces infiltration of mononuclear cells into the CNS when administered in vivo. Mononuclear cells purified from the central nervous system of control and compound 5J-4-treated mice were counted (top) or examined for numbers of CD4+ T cells (bottom) (n=4). *P<0.05, ***P<0.0005[1].Kim KD, et al. Calcium signaling via Orai1 is essential for induction of the nuclear orphan receptor pathway to drive Th17 differentiation. J Immunol. 2014 Jan 1;192(1):110-22.
  • Treatment with CRAC channel blockers reduces differentiation and expansion of TH17 cells (A) Compound 5J-4 treatment preferentially decreases TH17 differentiation in vivo. Expression levels of IFN-γ and IL-17A were measured in cells isolated from the draining lymph nodes (dLN) and CNS of DMSO and compound 5J-4-treated mice after 14 days of immunization with MOG35-55 peptide/CFA. Cells were stimulated with PMA and ionomycin and stained for CD4, IFN-γ, and IL-17A. Bar graphs on the right show average ± s.d.m. of indicated population of T cells from four mice. (B) Suppression of expansion and maintenance of pre-differentiated human TH17 cells by inhibition of CRAC channel activity. Human PBMCs were harvested, stimulated with anti-CD3 and anti-CD28 antibodies, and cultured with IL-1β and IL-23 in the presence of DMSO or 20 μM of compound 5D. Cells were restimulated with PMA and ionomycin at day 6 and examined for IL-17A and IFN-γ production. The graph shows results from eight donors.[1].Kim KD, et al. Calcium signaling via Orai1 is essential for induction of the nuclear orphan receptor pathway to drive Th17 differentiation. J Immunol. 2014 Jan 1;192(1):110-22.
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