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AS1269574

Alias: AS1269574; AS-1269574; AS 1269574
Cat No.:V11653 Purity: ≥98%
AS1269574 is a potent, orally bioactive GPR119 agonist with EC50 of 2.5 μM in HEK293 cells expressing human GPR119.
AS1269574
AS1269574 Chemical Structure CAS No.: 330981-72-1
Product category: GPR
This product is for research use only, not for human use. We do not sell to patients.
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Product Description
AS1269574 is a potent, orally bioactive GPR119 agonist with EC50 of 2.5 μM in HEK293 cells expressing human GPR119. AS1269574 activates the TRPA1 cation channel and stimulates glucagon-like peptide-1 (GLP-1) secretion. AS1269574 specifically induces glucose-dependent insulin secretion from pancreatic beta cells only under high glucose conditions. AS1269574 may be utilized in type 2 diabetes research.
Biological Activity I Assay Protocols (From Reference)
ln Vivo
AS1269574 significantly lowered blood glucose AUC 2 hours after injection (AUC0-2h)[1].
Animal Protocol
Animal/Disease Models: 8weeks old ICR mice [1]
Doses: 100 mg/kg
Route of Administration: Po
Experimental Results: Blood glucose AUC was Dramatically diminished after 2 hrs (hrs (hours)) of administration (AUC0-2h). Similarly, mouse plasma insulin AUC0-2h was Dramatically higher.
References

[1]. Identification of a novel GPR119 agonist, AS1269574, with in vitro and in vivo glucose-stimulated insulin secretion. Biochem Biophys Res Commun. 2010;400(3):437-441.

[2]. GPR119 Agonist AS1269574 Activates TRPA1 Cation Channels to Stimulate GLP-1 Secretion. Mol Endocrinol. 2016;30(6):614-629.

Additional Infomation
AS-1269574 is an organobromine compound.
These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C13H14BRN3O
Molecular Weight
308.173761844635
Exact Mass
307.032
Elemental Analysis
C, 50.67; H, 4.58; Br, 25.93; N, 13.64; O, 5.19
CAS #
330981-72-1
PubChem CID
5332859
Appearance
Light yellow to yellow solid powder
Density
1.5±0.1 g/cm3
Boiling Point
386.9±42.0 °C at 760 mmHg
Flash Point
187.8±27.9 °C
Vapour Pressure
0.0±0.9 mmHg at 25°C
Index of Refraction
1.648
LogP
2.32
Hydrogen Bond Donor Count
2
Hydrogen Bond Acceptor Count
4
Rotatable Bond Count
4
Heavy Atom Count
18
Complexity
246
Defined Atom Stereocenter Count
0
SMILES
CC1=CC(=NCCO)N=C(C2=CC=C(C=C2)Br)N1
InChi Key
DUKPGOOUJNUIOI-UHFFFAOYSA-N
InChi Code
InChI=1S/C13H14BrN3O/c1-9-8-12(15-6-7-18)17-13(16-9)10-2-4-11(14)5-3-10/h2-5,8,18H,6-7H2,1H3,(H,15,16,17)
Chemical Name
2-[[2-(4-bromophenyl)-6-methylpyrimidin-4-yl]amino]ethanol
Synonyms
AS1269574; AS-1269574; AS 1269574
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: This product requires protection from light (avoid light exposure) during transportation and storage.
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 (~811.24 mM)
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.08 mg/mL (6.75 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 (6.75 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 (6.75 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 3.2450 mL 16.2248 mL 32.4496 mL
5 mM 0.6490 mL 3.2450 mL 6.4899 mL
10 mM 0.3245 mL 1.6225 mL 3.2450 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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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.

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
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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
  • AS1269574 stimulates GLP-1 release and increases [Ca2+]i in STC-1 cells. A, AS1269574 (AS) exerted a dose-dependent action to stimulate the release of immunoreactive GLP-1 from STC-1 cells in static incubation assays. Data are averaged from five independent assays *, P < .02 (paired t test; see Materials and Methods for experimental details). B, AS1269574 but not the control vehicle solution containing 0.1% dimethylsulfoxide (vehicle) exerted a dose-dependent action to increase [Ca2+]i in fura-2 loaded STC-1 cell monolayers. A vertical arrow indicates when the test solutions were administered to individual wells of a 96-well plate, here and in all subsequent figures. Concentrations of test substances are the final concentrations present in the individual wells after injection of the test solutions, here and in all subsequent figures. C, The maximal increase of [Ca2+]i for a range of AS1269574 concentrations, as derived from the raw data presented in panel B. D, AS1269574 (AS; 30 μM) stimulated oscillations of [Ca2+]i in a single fura-2 loaded STC-1 cell. The horizontal bar indicates the time course during which AS1269574 was administered via a micropipette positioned immediately adjacent to the cell. B–D, For all examples depicted here, the findings are representative of a single experiment that was repeated a minimum of five times on five different occasions with similar results. E1–E3, Pseudocolor radiometric images of fura-2 fluorescence in clusters of STC-1 cells in which blue represents a low [Ca2+]I; green-yellow indicates an intermediate [Ca2+]i, and red indicates a high [Ca2+]i. Panels in E1–E3 indicate levels of [Ca2+]i before, during, and after application of AS1269574 (100 μM). These individual images were derived from the accompanying Supplemental Movie.[2]. GPR119 Agonist AS1269574 Activates TRPA1 Cation Channels to Stimulate GLP-1 Secretion. Mol Endocrinol. 2016;30(6):614-629.
  • GPR119-independent actions of AS1269574 to increase [Ca2+]i but not cAMP in STC-1 cell monolayers. A, GPR119 agonists AR231453 (AR; 6 μM), and OEA (30 μM) failed to increase [Ca2+]i, whereas AS1269574 (AS; 100 μM) did so. B, GPR119 antagonist TM43718 failed to significantly alter the action of AS1269574 to increase [Ca2+]i. C, AR231453 (1 μM) failed to increase [Ca2+]i under standard assay conditions (AR 1), whereas AS1269574 (AS; 100 μM) did so. Pretreatment with AR231453 (1 μM) for 15 minutes (AR pre, AS) failed to alter the subsequent action of AS1269574 (100 μM). D, A two-step injection mode assay revealed that combined administration of forskolin (F; 2 μM) and IBMX (I; 100 μM) failed to increase [Ca2+]i (FI, arrow 1, black trace), nor did the SES vehicle solution have any subsequent effect (SES, arrow 2, black trace). In the absence of forskolin, an initial injection of SES vehicle solution was without effect (SES, arrow 1, blue trace), whereas AS1269574 (100 μM) stimulated an increase of [Ca2+]i (AS 100, arrow 2, blue trace). When forskolin and IBMX were administered together during the initial injection (FI, arrow 1, red trace), there was no increase of [Ca2+]i, and the subsequent action of AS1269574 (AS 100, arrow 2, red trace) was not modified. E and F, AS1269574 (30 or 100 μM) failed to raise levels of cAMP, as inferred from its failure to induce an increase of the 535:484 nm AKAR3 FRET ratio (E). However, an increase of the 534:484 nm AKAR3 FRET ratio was measured in response to the positive control test solution containing forskolin (Fsk; 2 μM) and IBMX (100 μM). A–F, For all examples depicted here, the findings are representative of a single experiment that was repeated a minimum of three times on three different occasions with similar results.[2]. GPR119 Agonist AS1269574 Activates TRPA1 Cation Channels to Stimulate GLP-1 Secretion. Mol Endocrinol. 2016;30(6):614-629.
  • AS1269574 and AITC activate inward currents to increase [Ca2+]i in STC-1 cells. A, AS1269574 (AS 100) was applied at a concentration of 100 μM for 5 seconds directly to a single STC-1 cell (vertical arrows). The experiment was performed under conditions of whole-cell voltage clamp (−60 mV pipette holding potential) in which the cell was loaded with fura-2. AS1269574 activated an inward membrane current (A1), the time course of which closely matched the accompanying increase of [Ca2+]i (A2) in the same cell. Nearly identical findings were obtained in three separate experiments using a total of 10 cells. B, To determine the I-V relationship of the membrane current activated by AS1269574, the pipette potential was shifted as a voltage ramp from −70 mV to +70 mV (1 Vs−1). The baseline ramp current measured in the absence of AS1269574 was subtracted from the ramp current measured in the presence of AS1269574 so that the difference current could be calculated. The difference current is plotted as a function of the pipette potential in order to obtain the I-V relationship that indicates outward rectification characteristic of TRPA1 channels. For the illustrated cell, the reversal potential for the ramp current was +9 mV. For a total of 12 cells studied, the mean value of the reversal potential was +4 ± 2 mV (n = 12). C, RT-PCR detection of GPR119 and TRPA1 mRNA in STC-1 cells (reverse transcriptase [RT]). D, Representative example of the membrane current activated by a 5-second application of AITC (100 μM) to an STC-1 cell using methods described for panel A, above. Nearly identical findings were obtained in two separate experiments using a total of eight cells. E, Representative example of the I-V relationship for the membrane current activated by AITC. Note the close correspondence of this I-V relationship to that of panel B for AS1269574. For the illustrated cell, the reversal potential for the ramp current was +10 mV. For a total of 12 cells studied, the mean value of the reversal potential was +11 ± 2 mV (n = 12).[2]. GPR119 Agonist AS1269574 Activates TRPA1 Cation Channels to Stimulate GLP-1 Secretion. Mol Endocrinol. 2016;30(6):614-629.
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