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FPH2 (BRD-9424)

Alias:
Cat No.:V1930 Purity: ≥98%
FPH2 (also known as BRD-9424) is able to promote differentiation of iPS-derived hepatocytes.
FPH2 (BRD-9424)
FPH2 (BRD-9424) Chemical Structure CAS No.: 957485-64-2
Product category: Others
This product is for research use only, not for human use. We do not sell to patients.
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Purity & Quality Control Documentation

Purity: ≥98%

Product Description

FPH2 (also known as BRD-9424) is able to promote differentiation of iPS-derived hepatocytes. In vitro, FPH2 induced functional proliferation of hepatocytes and might be useful for expanding mature human primary hepatocytes. In human primary hepatocytes, FPH2 increased the number of nuclei undergoing mitosis and hepatocyte nuclei count in a concentration dependent way. Also, FPH2 (40 μM) increased the area of hepatocyte colonies with more hepatocytes and Ki67-positive nuclei, which exhibited hepatocyte nuclear morphologies. In primary human hepatocytes from six additional cell sources, FPH2 increased hepatocytes expansion. FPH2 increased hepatocyte at a rate that is consistent with liver regeneration kinetics in vivo.

Biological Activity I Assay Protocols (From Reference)
ln Vitro
FPH2 may be helpful for growing mature human primary hepatocytes since it causes functional proliferation of hepatocytes in vitro. During primary screening, FPH1 and FPH2 can increase the number of hepatocyte nuclei and/or increase the number of nuclei undergoing mitosis. These effects on hepatocytes are concentration dependant. FPH1 and FPH2-treated cells continue to perform liver-specific tasks. FPH2 doubles the rate of hepatocyte induction over 7 days, which is in line with the kinetics of liver regeneration that have been documented in vivo[1].
ln Vivo
FPH2 increased hepatocyte at a rate that is consistent with liver regeneration kinetics in vivo
Animal Protocol


References
[1]. Shan J, et al. Identification of small molecules for human hepatocyte expansion and iPS differentiation. Nat Chem Biol. 2013 Aug;9(8):514-20.
Additional Infomation
4-[[(5-chloro-2-methoxyanilino)-sulfanylidenemethyl]amino]-1-ethyl-3-pyrazolecarboxamide is a member of thioureas.
These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C14H16CLN5O2S
Molecular Weight
353.83
Exact Mass
353.071
CAS #
957485-64-2
Related CAS #
957485-64-2
PubChem CID
2208391
Appearance
Typically exists as solid at room temperature
Density
1.45±0.1 g/cm3
LogP
3.319
Hydrogen Bond Donor Count
3
Hydrogen Bond Acceptor Count
4
Rotatable Bond Count
5
Heavy Atom Count
23
Complexity
441
Defined Atom Stereocenter Count
0
SMILES
O=C(C1C(NC(NC2C(OC)=CC=C(Cl)C=2)=S)=CN(CC)N=1)N
InChi Key
PCHRYHSDDPPZBV-UHFFFAOYSA-N
InChi Code
InChI=1S/C14H16ClN5O2S/c1-3-20-7-10(12(19-20)13(16)21)18-14(23)17-9-6-8(15)4-5-11(9)22-2/h4-7H,3H2,1-2H3,(H2,16,21)(H2,17,18,23)
Chemical Name
4-[[[(5-chloro-2-methoxyphenyl)amino]thioxomethyl]amino]-1-ethyl-1H-pyrazole-3-carboxamide
Synonyms

FPH2, FPH-2, FPH 2; BRD-9424, BRD9424, BRD 9424

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)
DMSO:70 mg/mL (197.8 mM)
Water:<1 mg/mL
Ethanol:<1 mg/mL
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.07 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

Solubility in Formulation 2: ≥ 2.5 mg/mL (7.07 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 2.8262 mL 14.1311 mL 28.2622 mL
5 mM 0.5652 mL 2.8262 mL 5.6524 mL
10 mM 0.2826 mL 1.4131 mL 2.8262 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.

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

Clinical Trial Information
NCT Number Recruitment interventions Conditions Sponsor/Collaborators Start Date Phases
NCT05482594 RECRUITING Diagnostic Test:ELISA
Diagnostic Test:Videocapillaroscopy
Systemic Sclerosis Brugmann University Hospital 2022-01-11
NCT00434239 UNKNOWN STATUS Drug:Lenalidomide+Ancestim Myelodysplasia Peter MacCallum Cancer
Centre, Australia
2007-02 Early Phase 1
NCT00001398 COMPLETED Drug:Recombinant Methionyl Huma
Stem Cell Factor(r-metHuSCF)
Aplastic Anemia
Pancytopenia
National Heart,Lung,
Blood Institute(NHLBI)
1993-10 Phase 1
NCT00005783 COMPLETED Drug:Recombinant-methionyl
human stem cell factor
Hemoglobin SC Disease
Sickle Cell Anemia
National Institute of Diabetes
Digestive and Kidney Diseases (NIDDK)
2000-03 Phase 1
NCT00058045 COMPLETED Biological:aldesleukin
Biological:recombinant human
stem cell factor
Lymphoma Roswell Park Cancer Institute 2002-08 Phase 1
Biological Data
  • High-throughput identification of small molecules that induce proliferation and enhance functions of primary human hepatocytes (a) High-throughput liver platform. Primary human hepatocytes (green) were seeded on a feeder layer of confluent J2-3T3 fibroblasts (red) in 384-well plates. Line graph shows representative rate of albumin secretion in screening co-cultures and hepatocyte-only cultures (green) over time. Bar graph displays albumin secretion as a function of hepatocyte density in screening cultures. Upper inset phase-contrast imaging shows morphology of feeder-layer co-cultures (scale bar = 100 µm). Far right, Hoechst staining of screening co-cultures shows that hepatocyte nuclei (green *) have uniform texture while fibroblast nuclei (red *) are punctate (scale bar = 50 µm). Automated high-content imaging assay identifies and classifies individual nuclei. (b) Chemical screening. Bar graph depicts categories of screened and hit compounds. Scatter plots display replicates of the screen, shown separately for the image-based proliferation and competitive-ELISA functional readouts. Blue and red data points represent DMSO and experimental small molecules, respectively. Boxed regions indicate hit zones. (c) Hit validation. All primary hits were retested in an 8-point dose-response curve. Active hits had increasing curves of hepatocyte nuclei counts and/or decreasing curves of competitive [Albumin] with flat curves of cell-free [Albumin]. (d) Chemical structures of hit compounds FPH1, FPH2 and FH1. All data presented as mean ± s.d. Nat Chem Biol . 2013 Aug;9(8):514-20.
  • Expansion of primary human hepatocytes (a) Primary screening data for FPH1 and FPH2. Data presented as mean ± s.d. (b) Ki67 (red) and albumin (green) immunofluorescent staining after 6 days of culture. Bar graphs show CellProfiler quantification of displayed images. (c) FACS (right) and Cellometer (left) Automated Cell Counter analysis. Fibroblasts were labeled with CM-DiI prior to initiation of culture in order to allow identification of hepatocytes via negative selection. FACS cell counting was further enabled by fluorescent counting beads. Control cultures were treated with empty vehicle (DMSO). Data presented as mean ± SEM. Nat Chem Biol . 2013 Aug;9(8):514-20.
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