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PU-02

Alias: PU02; PU 02; PU-02
Cat No.:V13364 Purity: ≥98%
PU02 is an analogue of 6-MP and a negative allosteric modulator (NAM) of 5-HT3 receptors.
PU-02
PU-02 Chemical Structure CAS No.: 313984-77-9
Product category: New1
This product is for research use only, not for human use. We do not sell to patients.
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Product Description
PU02 is an analogue of 6-MP and a negative allosteric modulator (NAM) of 5-HT3 receptors. Its IC50 was measured in HEK293 cells after transfection with 5-HT3A and 5-HT3AB receptors. 0.36 and 0.73 μM respectively.
Biological Activity I Assay Protocols (From Reference)
ln Vitro
HepG2 cell viability steadily declines in response to PU02 (NMMP) (0-200 μM) (IC50=48.585 μM). PU02 (NMMP) was less hazardous to L02 cells than 6-MP after 48 hours of treatment [2]. The viability of several examined cell lines, including SMMC-7721, MDA-MB-231, RKO, and HCT-8 cells, was inhibited by PU02 (NMMP) at 6.25 or 25 μM [2]. In the G2/M phase, PU02 (NMMP) causes cell cycle arrest. HepG2 cells' expression of CDK4 and cyclin B1/D1 is time-dependently down-regulated by PU02 (NMMP), while cyclin E expression is unaffected [2]. Caspases-3 cleavage was significantly increased in PU02 (NMMP)-treated cells, suggesting increased apoptotic activity [2].
Cell Assay
Apoptosis analysis [2]
Cell Types: HepG2 cells.
Tested Concentrations: 6.26, 12.5, 25, 50 μM.
Incubation Duration: 6, 12, 24, 36 hrs (hours).
Experimental Results: Mitochondria-dependent apoptosis was induced.
References

[1]. Discovery of a novel allosteric modulator of 5-HT3 receptors: inhibition and potentiation of Cys-loop receptor signaling through a conserved transmembrane intersubunit site. J Biol Chem. 2012 Jul 20;287(30):25241-54.

[2]. 6-[(1-naphthylmethyl)sulfanyl]-9H-purine induces G2/M phase arrest and apoptosis in human hepatocellular carcinoma HepG2 cells. Eur J Pharmacol. 2012 Nov 15;695(1-3):27-33.

These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C16H12N4S
Molecular Weight
292.36
Exact Mass
292.078
CAS #
313984-77-9
PubChem CID
720937
Appearance
White to off-white solid powder
LogP
3.798
Hydrogen Bond Donor Count
1
Hydrogen Bond Acceptor Count
4
Rotatable Bond Count
3
Heavy Atom Count
21
Complexity
353
Defined Atom Stereocenter Count
0
InChi Key
BGMSTNYJYPSLHN-UHFFFAOYSA-N
InChi Code
InChI=1S/C16H12N4S/c1-2-7-13-11(4-1)5-3-6-12(13)8-21-16-14-15(18-9-17-14)19-10-20-16/h1-7,9-10H,8H2,(H,17,18,19,20)
Chemical Name
6-(naphthalen-1-ylmethylsulfanyl)-7H-purine
Synonyms
PU02; PU 02; PU-02
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

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.
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 : ~250 mg/mL (~855.11 mM)
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.08 mg/mL (7.11 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 20.8 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: ≥ 2.08 mg/mL (7.11 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
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.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (7.11 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 20.8 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 3.4204 mL 17.1022 mL 34.2044 mL
5 mM 0.6841 mL 3.4204 mL 6.8409 mL
10 mM 0.3420 mL 1.7102 mL 3.4204 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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Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
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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.

Biological Data
  • Mechanism of action of PU02 at the human 5-HT3A receptor. A, concentration-inhibition curves for tropisetron, PU01, and PU02 in the [3H]GR65630 competition binding assay to 5-HT3A-HEK293 cell membranes. Tropisetron inhibited [3H]GR65630 binding to the receptor with a pKi ± S.E. value of 9.59 ± 0.18 (n = 3). B, concentration-response curves of 5-HT at 5-HT3A-HEK293 cells in the absence or presence of five different concentrations of PU02 in the FMP assay. No significant response was observed upon application of 5-HT concentrations up to 100 μm when PU02 was present in the assay at concentrations of 20 μm and above (not shown). C, concentration-response curves for 5-HT at 5-HT3A-HEK293 cells in the FMP assay after preincubation of the cells with Krebs buffer or with Krebs buffer supplemented with 30 μm PU02 or 10 nm ondansetron for 30 min followed by three rounds of washes with Krebs buffer. The experiments were performed as described under “Experimental Procedures,” and the figures depict data ± S.D. (error bars) of duplicate determinations from representative experiments (n = 3–4). FU, fluorescence units.[1].Discovery of a novel allosteric modulator of 5-HT3 receptors: inhibition and potentiation of Cys-loop receptor signaling through a conserved transmembrane intersubunit site. J Biol Chem. 2012 Jul 20;287(30):25241-54.
  • Functional properties of PU02 at 5-HT3Rs and nAChRs. A, chemical structure of PU02 and concentration-inhibition curves for PU02 at human 5-HT3A-HEK293 cells in the FMP assay (■) and at human 5-HT3A- (●) and 5-HT3AB-HEK293 cells (○) in the Ca2+/Fluo-4 assay. B, concentration-inhibition curves for PU02 at mouse α4β2-HEK293T, rat α3β4-HEK293, and human α7-GH3 cells in the FMP (α4β2, α3β4) and Ca2+/Fluo-4 (α7) assays. The assays were performed as described under “Experimental Procedures,” using EC80 agonist concentrations, and the results shown represent data ± S.D. (error bars) of duplicate determinations from single representative experiments (n = 3–5).[1].Discovery of a novel allosteric modulator of 5-HT3 receptors: inhibition and potentiation of Cys-loop receptor signaling through a conserved transmembrane intersubunit site. J Biol Chem. 2012 Jul 20;287(30):25241-54.
  • Electrophysiological characterization of PU02 at the 5-HT3A receptor expressed in COS-7 cells. A, currents induced by 20 μm 5-HT (∼EC70–80) pulses (solid bar, 45-s interval) were inhibited by PU02 (dotted bar). Scale bars, 200 pA (vertical) and 5 s (horizontal). Peak current amplitudes induced by 20 μm 5-HT in the presence of PU02 were base line-subtracted and normalized to those in absence of antagonist. B, concentration-inhibition curve for PU02. Data are given as mean ± S.E. values (error bars) based on 5–6 cells on at least three different days and transfections. C, association (current rise) and desensitization (current decay) time constants (τ values) for the 5-HT response in the absence or presence of 0.3 μm PU02 were calculated by fitting either phase of selected currents to applicable single exponential equations.[1].Discovery of a novel allosteric modulator of 5-HT3 receptors: inhibition and potentiation of Cys-loop receptor signaling through a conserved transmembrane intersubunit site. J Biol Chem. 2012 Jul 20;287(30):25241-54.
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