Reversine

Alias: Reversine

Cat No.:V0351 Purity: ≥98%

COA Product Data Instructions for use SDS

Reversine is a cell-permeable, selective and ATP-competitive inhibitor of human A3 adenosine receptor and a pan-aurora A/B/C kinase inhibitor as well with potential anticancer activity.

Reversine Chemical Structure CAS No.: 656820-32-5
Product category: Adenosine Receptor

This product is for research use only, not for human use. We do not sell to patients.

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Citations by Top Journals: Nature, Cell, Science, etc.

Top Publications Citing lnvivochem Products

Nature 2021, 597(7874):119-125.

Cell 2020,183:1202-1218.e25.

Science Adv 2022: 8, eabm9427.

Nature 597, 119–125 (2021)

Cell Stem Cell 2020,26(6):845-861.e12.

Science Trans Med 2023,15(701):eadd7872.

STTT 2023,8(1):91.

Cell Reports 2023,42(4):112313

Science Trans Med 2023, 15: eadd7872.

Cancer Cell 2021,39(4):529-547.e7.

Cancer Discov 2022,12(4):1106-1127.

Immunity 2023,56(3):620-634.e11.

J Am Chem Soc 2022,144:21831-21836.

Cell 2020,183:1202-1218.e25.

Science Adv 2022:8,eabm9427.

Nature 2021,597(7874):119-125.

Cell 2020,183:1202-1218.e25.

PNAS Nexus 2022,1(3):pgac063.

ACS Nano 2023,17(10):9110-9125.

Biomaterial 2023 Sep16:302:122333.

Purity & Quality Control Documentation

Current batch：

Purity: ≥98%

COA

MSDS

NMR

Instructions

Product Description
Bioactivity & Protocols
Physicochemical Properties
Solubility Data
Calculators
Biological Data

Product Description

Reversine is a cell-permeable, selective and ATP-competitive inhibitor of human A3 adenosine receptor and a pan-aurora A/B/C kinase inhibitor as well with potential anticancer activity. It inhibits pan-aurora A/B/C kinase with IC50s of 12 nM/13 nM/20 nM and the human A3 adenosine receptor with a Ki of 0.66 μM, respectively. It exhibits strong antiproliferative properties in vitro and strong antitumor efficaciousness in vivo.

Biological Activity I Assay Protocols (From Reference)

Targets

Aurora A ( IC50 = 400 nM ); Aurora B ( IC50 = 500 nM ); Aurora C ( IC50 = 400 nM )

ln Vitro

In vitro activity: Reversine causes cells committed to the myogenic lineage to develop into multipotent mesenchymal progenitor cells, which multiply and redifferentiate into bone and adipose tissues.[1] In stable transfected Chinese hamster ovary (CHO) cells, reversine, an A3 adenosine receptor antagonist, competitively inhibits the production of cAMP stimulated by forskolin.[2] In HCT116 cells, reversine prevents the phosphorylation of histone H3, a well-known Aurora target. Reversine also induces cell death and potently inhibits the proliferation of several tumor cell types. In primary human tumor samples, Reversine also prevents leukemic cells from forming colonies.[3] Reversine and aspirin together have a synergistic effect on cervical cancer cell growth inhibition and apoptosis induction. [4]

ln Vivo

Reversine (10 mg/kg i.p.) and aspirin, when compared to the control agents, result in greater reductions in tumor weight and volume in mice inoculated with U14 tumors.[4]

Enzyme Assay

In the A3 AR competitive binding assay, each tube has 50 μL of [¹²⁵I]4-amino-3-iodobenzyl)adenosine-5′-N-methyluronamide (0.5 nM), 100 μL of membrane suspension (20 μg of protein), and 50 μL of increasing concentrations of the test ligands in Tris-HCl buffer (50 mM, pH 7.4) containing 10 mM MgCl₂ and 1 mM EDTA. The buffer containing 10 mM 5′-N-ethylcarboxamidoadenosine is used to measure nonspecific binding. For sixty minutes, the mixtures are incubated at 25°C. Using an MT-24 cell harvester, binding reactions are stopped by filtering through Whatman GF/B filters while operating at a lower pressure. 9 milliliters of ice-cold buffer are used to wash filters three times. A Beckman γ-counter is used to measure radioactivity, and the percentage of inhibition is computed.

Cell Assay

Viability of various tumor cell lines is evaluated by means of ATPlite 1step. In short, a crescent amount of reversine is present when 2 × 10⁴ cells are plated in a 96-well plate for each well. 100 μL of ATPlite solution is added to each well of the recovered plates after a 72-hour period. A luminescence measurement is made using an EnVision Multilabel plate reader after the plates are shaken for two minutes at 700 rpm. We analyze each sample three times.

Animal Protocol

Mice: female athymic The mice used are 6–8 week old BALB/c nude mice. Subcutaneous injection of a 5x10⁶ cell suspension in 100 μL of RPMI-1640 medium is performed using U14 cell suspension. The goal of cervical tumor development is to produce histologically intact tumors for medication therapy. Twenty-five of these mice were randomized into one of five groups based on random assignment once the tumors reached a diameter of approximately 1 cm. The groups included mice treated with RPMI-1640 medium, mice treated with DMSO, mice treated with Reversine (10 mg/kg), mice treated with aspirin (1 μg/kg), and mice treated with a combination of Reversine and aspirin. Three times a week, the body weight and the size of the tumor at the injection site are measured. Every three days, the size of a tumor is measured from two diameters, and the formula L×S2/2 is used to estimate the tumor volume (where L represents the longest diameter and S the shortest).

References

[1]. J Am Chem Soc . 2004 Jan 21;126(2):410-1.

[2]. J Med Chem . 2005 Jul 28;48(15):4910-8.

[3]. Mol Cancer Ther . 2008 May;7(5):1140-9.

[4]. Cytotechnology . 2013 Aug;65(4):643-53.

These protocols are for reference only. InvivoChem does not independently validate these methods.

Physicochemical Properties

Molecular Formula

C21H27N7O

Molecular Weight

393.23

Exact Mass

393.23

Elemental Analysis

C, 64.10; H, 6.92; N, 24.92; O, 4.07

CAS #

656820-32-5

Related CAS #

656820-32-5

Appearance

White solid powder

SMILES

C1CCC(CC1)NC2=NC(=NC3=C2NC=N3)NC4=CC=C(C=C4)N5CCOCC5

InChi Key

ZFLJHSQHILSNCM-UHFFFAOYSA-N

InChi Code

InChI=1S/C21H27N7O/c1-2-4-15(5-3-1)24-20-18-19(23-14-22-18)26-21(27-20)25-16-6-8-17(9-7-16)28-10-12-29-13-11-28/h6-9,14-15H,1-5,10-13H2,(H3,22,23,24,25,26,27)

Chemical Name

6-N-cyclohexyl-2-N-(4-morpholin-4-ylphenyl)-7H-purine-2,6-diamine

Synonyms

Reversine

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: 5~20 mg/mL (12.7~50.8 mM)

Water: <1 mg/mL

Ethanol: <1 mg/mL

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.5430 mL	12.7152 mL	25.4304 mL
	5 mM	0.5086 mL	2.5430 mL	5.0861 mL
	10 mM	0.2543 mL	1.2715 mL	2.5430 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

Mass

Concentration

Volume

Molecular Weight*

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

Calculate the Mass of a compound required to prepare a solution of known volume and concentration
Calculate the Volume of solution required to dissolve a compound of known mass to a desired concentration
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See Example

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.

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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)
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The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box

Molecular formula

Total molecular weight

g/mol

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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
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
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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)

Dosage mg/kg

Average weight of animals g

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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.

Note： (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

Inhibition of growth of cervical cancer cells by reversine and aspirin a inhibition rate of different drug concentrations on HeLa cells treated with reversine at 10 μmol/L increased compared with mock group, P < 0.05, however, inhibition rate of aspirin group at 10 mmol/L increased significantly, P < 0.05. b reversine (10 μmol/L), aspirin (10 mmol/L) and their combination on U14 cells with different time treatment, inhibition rate of combination group decreased significantly similar to HeLa cells compared with the drug used separately, P < 0.05. c Inhibition rate of the combination of both agents on HeLa and U14 cells at different processing times. Cytotechnology . 2013 Aug;65(4):643-53.

Cell cycle and mitochondria potential of HeLa and U14 cells a cell cycle analysis of HeLa cells after treatment with reversine, aspirin or a combination of both (10 μmol/L reversine, 10 mmol/L aspirin). Cytotechnology . 2013 Aug;65(4):643-53.

Soft agar colony formation analysis and expression of cell cycle and apoptosis related genes in HeLa or U14 cells after treatment with reversine and aspirin for 48 h a HeLa and U14 cells colony formation decreased significantly in the combination group. Cytotechnology . 2013 Aug;65(4):643-53.