R 59949

Alias: R-59949 R59949 R 59949

Cat No.:V11333 Purity: ≥98%

COA Product Data Instructions for use SDS

R59949 is a pan-diacylglycerol kinase (DGK) inhibitor (antagonist) with IC50 of 300 nM.

R 59949 Chemical Structure CAS No.: 120166-69-0
Product category: PKC

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.

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Nature 2024 Dec 18.

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

Cell 2020,183:1202-1218.e25.

Science Adv 2022: 8, eabm9427.

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Cell Stem Cell 2024, 31:106-126.e13.

Nature 597, 119–125 (2021)

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

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

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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.

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

Product Description

R59949 is a pan-diacylglycerol kinase (DGK) inhibitor (antagonist) with IC50 of 300 nM. R59949 strongly inhibits the activity of type I DGKα and γ, and moderately weakens the activity of type II DGKθ and κ. R59949 activates protein kinase C (PKC) by increasing levels of the endogenous ligand diacylglycerol.

Biological Activity I Assay Protocols (From Reference)

ln Vitro	Vasopressin- and collagen-induced release response and aggregation are markedly enhanced in the presence of R59949, which is linked to the production of arachidonic acid metabolites [1]. R59949 restricts the import of CCL2 in THP-1 monocytes. By decreasing the concentration of L-arginine solution across the plasma membrane in vascular smooth muscle cells, R59949 adsorbs the generation of inducible nitric oxide [4]. Half maximal concentration of the Ca2+ signal is 8.6 μM[2].
References	[1]. The role of endogenously formed diacylglycerol in the propagation and termination of platelet activation. A biochemical and functional analysis using the novel diacylglycerol kinase inhibitor, R 59 949. J Biol Chem. 1989;264(6):3274-3285. [2]. Inhibitors of DAG metabolism suppress CCR2 signalling in human monocytes. Br J Pharmacol. 2019;176(15):2736-2749. [3]. Effect of novel modulators of protein kinase C activity upon chemotherapy-induced differentiation and apoptosis in myeloid leukemic cells. Anticancer Drugs. 2002;13(7):725-733. [4]. R59949, a diacylglycerol kinase inhibitor, inhibits inducible nitric oxide production through decreasing transplasmalemmal L-arginine uptake in vascular smooth muscle cells. Naunyn Schmiedebergs Arch Pharmacol. 2017;390(2):207-214.
Additional Infomation	3-[2-[4-[bis(4-fluorophenyl)methylidene]-1-piperidinyl]ethyl]-2-sulfanylidene-1H-quinazolin-4-one is a diarylmethane.

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

Physicochemical Properties

Molecular Formula	C28H25F2N3OS
Molecular Weight	489.58
Exact Mass	489.169
CAS #	120166-69-0
PubChem CID	657356
Appearance	White to off-white solid powder
Density	1.36g/cm3
Boiling Point	637.1ºC at 760mmHg
Flash Point	339.1ºC
Vapour Pressure	3.94E-16mmHg at 25°C
Index of Refraction	1.688
LogP	5.873
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	5
Heavy Atom Count	35
Complexity	770
Defined Atom Stereocenter Count	0
InChi Key	ZCNBZFRECRPCKU-UHFFFAOYSA-N
InChi Code	InChI=1S/C28H25F2N3OS/c29-22-9-5-19(6-10-22)26(20-7-11-23(30)12-8-20)21-13-15-32(16-14-21)17-18-33-27(34)24-3-1-2-4-25(24)31-28(33)35/h1-12H,13-18H2,(H,31,35)
Chemical Name	3-(2-(4-(bis(4-fluorophenyl)methylene)piperidin-1-yl)ethyl)-2-thioxo-2,3-dihydroquinazolin-4(1H)-one
Synonyms	R-59949 R59949 R 59949
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 : ~100 mg/mL (~204.26 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.11 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 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	2.0426 mL	10.2128 mL	20.4257 mL
	5 mM	0.4085 mL	2.0426 mL	4.0851 mL
	10 mM	0.2043 mL	1.0213 mL	2.0426 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

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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?

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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:

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

Molecular formula

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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:

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)
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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)

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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

Inhibitors of DAG kinase and DAG lipase attenuate CCL2‐evoked Ca2+ signalling and migration in THP‐1 cells. (a) Effect of DAG kinase inhibitor R59949 (30 μM) on Ca2+ responses evoked by CCL2 (50 ng·ml−1; n = 7). Averaged responses are shown in the presence of vehicle (closed circles) and inhibitor (open circles). (b) R59949 concentration–inhibition curve (IC50 = 9 ± 1 μM; n = 7) against Ca2+ responses evoked by CCL2 (50 ng·ml−1). (c) Effect of 30‐μM R59949 on CCL2 concentration–response curve (n = 7). (d) Effect of DAG lipase inhibitor RHC80267 (30 μM) on Ca2+ responses evoked by CCL2 (50 ng·ml−1; n = 7). Averaged responses are shown in the presence of vehicle (closed circles) and inhibitor (open circles). (e) RHC80267 concentration–inhibition curve (IC50 = 9 ± 1 μM; n = 7) against Ca2+ responses evoked by CCL2 (50 ng·ml−1). (f) Effect of 30‐μM RHC80267 on CCL2 concentration–response curve (n = 7). (g) Effect of R59949 (30 μM) and RHC80267 (30 μM) on THP‐1 transmigration to CCL2 (3 ng·ml−1). *P < .05 versus vehicle and # P < .05 versus CCL2 alone (n = 8). F ratio is the Ca2+ response as measured by the Fura‐2 emission intensity ratio when excited at 340 and 380 nm. Data in concentration–response/inhibition curves are expressed as a percentage of the control response in the presence of vehicle alone. [2]. Inhibitors of DAG metabolism suppress CCR2 signalling in human monocytes. Br J Pharmacol. 2019;176(15):2736-2749.

Responses to CCL2 but not fMLP are attenuated by DAG kinase and DAG lipase inhibitors in freshly isolated human monocytes. Effect of (a) DAG kinase inhibitor R59949 (30 μM) and (b) DAG lipase inhibitor RHC80267 (30 μM) on Ca2+ responses evoked by CCL2 (50 ng·ml−1) in monocytes (n = 8). Averaged responses are shown in the presence of vehicle (closed circles) and inhibitor (open circles). (c) Bar chart showing effect of R59949 (30 μM) and RHC80267 (30 μM) on the peak Ca2+ response evoked by CCL2 (50 ng·ml−1; n = 8). (d) Effect of R59949 (30 μM) and RHC80267 (30 μM) on freshly isolated monocyte transmigration to CCL2 (3 ng·ml−1). *P < .05 versus vehicle and # P < .05 versus CCL2 alone (n = 8). Lack of effect of (e) R59949 (30 μM) or (f) RHC80267 (30 μM) on Ca2+ responses evoked by fMLP (10 μM) in monocytes (n = 6). Averaged responses are shown in the presence of vehicle (closed circles) and inhibitor (open circles).[2]. Inhibitors of DAG metabolism suppress CCR2 signalling in human monocytes. Br J Pharmacol. 2019;176(15):2736-2749.

Inhibition of DAG kinase or DAG lipase does not reduce the cell surface population of CCR2 in freshly isolated monocytes and THP‐1 cells. Three representative (n = 6) flow cytometry profiles of freshly isolated CD14+ monocytes (a) and THP‐1 cells (b) labelled with anti‐CCR2 antibodies or isotype control. Cells were treated with vehicle control, R59949 (30 μM), or RHC80267 (30 μM). Anti‐CCR2 cell surface immunoreactivity is indistinguishable between vehicle control and test groups.[2]. Inhibitors of DAG metabolism suppress CCR2 signalling in human monocytes. Br J Pharmacol. 2019;176(15):2736-2749.