MRTX1133 formic
Based on 58 publication(s) in Google Scholar
MRTX1133 formic is a noncovalent, potent, and selective KRAS G12D inhibitor. MRTX1133 formic optimally fills the switch II pocket and extends three substituents to favorably interact with the protein, resulting in an estimated KD against KRAS G12D of 0.2 pM. MRTX1133 formic prevents SOS1-catalyzed nucleotide exchange and/or formation of the KRASG12D/GTP/RAF1 complex, thereby inhibiting mutant KRAS-dependent signal transduction. MRTX1133 formic shows efficacy in tumor models harboring KRAS G12D mutations.
Nos produits utilisent uniquement pour la recherche. Nous ne vendons pas aux patients.
- Formule: C34H33F3N6O4
- Masse moléculaire:646.66
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Stockage:
4°C, sealed storage, away from moisture
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
Publications Citing Use of MedChemExpress (MCE) MRTX1133 formic
More- Signal Transduct Target Ther. 2026 Jan 16;11(1):33. [Abstract]
- Nature. 2026 May;653(8113):254-264. [Abstract]
- Nature. 2026 Jan;649(8098):1022-1031. [Abstract]
- Cancer Cell. 2025 Jul 25:S1535-6108(25)00310-1. [Abstract]
- Immunity. 2023 Nov 14;56(11):2570-2583.e6. [Abstract]
- Cancer Discov. 2025 Oct 21. [Abstract]
- Cancer Discov. 2024 Dec 2;14(12):2430-2449. [Abstract]
- Cancer Discov. 2024 Oct 4;14(10):1964-1989. [Abstract]
- Nat Cancer. 2026 Jan;7(1):116-130. [Abstract]
- Cancer Res. 2026 Jul 2;86(13):3270-3286. [Abstract]
- Cancer Res. 2026 Apr 29:10.1158/0008-5472.CAN-24-1984. [Abstract]
- Cancer Res. 2025 Jan 2;85(1):101-117. [Abstract]
- Cancer Res. 2024 Nov 4;84(21):3629-3639. [Abstract]
- Nat Commun. 2026 Jun 15;17(1):5288. [Abstract]
- Nat Commun. 2026 Feb 24;17(1):3113. [Abstract]
- Nat Commun. 2025 Dec 15. [Abstract]
- Nat Commun. 2025 Feb 19;16(1):1765. [Abstract]
- J Adv Res. 2026 Feb 13:S2090-1232(26)00144-X. [Abstract]
- Adv Sci (Weinh). 2026 Feb 12:e15654. [Abstract]
- Cell Rep Med. 2026 Feb 17;7(2):102613. [Abstract]
- Cell Rep Med. 2025 Nov 18;6(11):102446. [Abstract]
- Cell Rep Med. 2025 Feb 18;6(2):101966. [Abstract]
- Sci Adv. 2024 Dec 13;10(50):eadq4274. [Abstract]
- Cell Death Dis. 2025 Dec 23. [Abstract]
- Cancer Lett. 2024 Jun 1:591:216904. [Abstract]
- Clin Cancer Res. 2026 Mar 9. [Abstract]
- Clin Cancer Res. 2024 Sep 13;30(18):4082-4099. [Abstract]
- Acta Pharmacol Sin. 2023 Jul;44(7):1475-1486. [Abstract]
- Proc Natl Acad Sci U S A. 2026 May 5;123(18):e2601788123. [Abstract]
- Proc Natl Acad Sci U S A. 2024 May 21;121(21):e2403685121. [Abstract]
- Mol Med. 2024 Nov 5;30(1):199. [Abstract]
- EMBO Mol Med. 2025 Jun;17(6):1355-1392. [Abstract]
- Cell Rep. 2026 Jun 23;45(6):117520. [Abstract]
- Cell Rep. 2026 May 4;45(5):117338. [Abstract]
- Cell Rep. 2024 Aug 22;43(9):114667. [Abstract]
- Mol Cancer Ther. 2025 Oct 23. [Abstract]
- ACS Appl Bio Mater. 2024 Dec 16;7(12):8489-8502. [Abstract]
- Sci Rep. 2026 Jan 7;16(1):4144. [Abstract]
- Transl Oncol. 2024 Dec 9:52:102235. [Abstract]
- Mol Oncol. 2025 Feb;19(2):295-310. [Abstract]
- ACS Pharmacol Transl Sci. 2024 Dec 2;7(12):3921-3934. [Abstract]
- STAR Protoc. 2026 Jun 26;7(3):104649. [Abstract]
- bioRxiv. 2026 May 9.
- bioRxiv. 2026 Apr 6.
- The Medical College of Wisconsin. 2026.
- bioRxiv. 2026 Apr 15.
- bioRxiv. 2026 Jan 5:2026.01.05.697773. [Abstract]
- medRxiv. 2025 Sep 12.
- bioRxiv. 2025 Sep 18:2025.09.16.676456. [Abstract]
- bioRxiv. 2025 Jul 30:2025.07.25.666783. [Abstract]
- bioRxiv. 2025 Jun 25:2025.06.20.660715. [Abstract]
- University of Kansas. 2025.
- bioRxiv. 2025 May 13:2025.05.10.653074. [Abstract]
- bioRxiv. 2025 January 14.
- bioRxiv. 2024 July 23.
- bioRxiv. 2023 Oct 20:2023.10.19.563158. [Abstract]
- bioRxiv. 2023 Oct 6.
- bioRxiv. 2023 Sep 17.
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RT-PCR
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WB
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RT-PCR
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WB
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WB
Activité biologique
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KRas G12D 0.2 pM (Kd) |
MRTX1133 formic inhibits ERK phosphorylation in the AGS cell line with an IC50 ranging 1-10 nM (AsPC-1, Panc 04.03, Panc 02.03, SW1990, GP2D, Suit2, A427, SNU1033, and HPAC cells). In a 2D viability assay, the IC50 of MRTX1133 formic is 6 nM against AGS cells (KRAS G12D), while demonstrating more than 500-fold selectivity against MKN1, a cell line which is dependent on KRAS for its growth and survival due to the amplification of wild-type KRAS[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:6-8-weekold, female, athymic nude-Foxn1nu mice (Panc 04.03 model)[1]
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Dosage:3, 10, or 30 mg/kg
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Administration:Intraperitoneal; twice a day for 28 days
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Result:Resulted in a dose-dependent antitumor activity with 94% growth inhibition observed at 3 mg/kg BID (IP) and tumor regressions of -62% and -73% observed at 10 and 30 mg/kg BID (IP), respectively.
| NCT Number | Sponsor | Condition | Start Date |
Phase
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|---|---|---|---|---|
| NCT01329991 | Plexxikon| | 2011-05 | PHASE1 |
Chemical Information
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Appearance Solid
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Masse moléculaire 646.66
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Formule C34H33F3N6O4
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Color Yellow to khaki
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SMILES
O=CO.OC1=CC(C2=C(F)C3=NC(OC[C@]45C[C@@H](F)CN4CCC5)=NC(N6C[C@H]7N[C@H](CC7)C6)=C3C=N2)=C8C(C#C)=C(F)C=CC8=C1
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Livraison
Room temperature in continental US; may vary elsewhere.
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Stockage
4°C, sealed storage, away from moisture
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
Publications (58)
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Journal Impact Factor
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Most Recent
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Signal Transduct Target Ther
Vertical RAS pathway inhibition in pancreatic cancer drives therapeutically exploitable mitochondrial alterations. [Abstract]2026 Jan 16;11(1):33. PMID: 41545339 -
Nature
2026 May;653(8113):254-264. PMID: 42020743 -
Nature
2026 Jan;649(8098):1022-1031. PMID: 41225001
MRTX1133 formic purchased from MedChemExpress. Usage Cited in: Nature. 2026 Jan;649(8098):1022-1031. [Abstract]
MRTX1133 (10, 100 nM; 24 h). KPC cells were treated for 24 h and analysed using quantitative PCR with reverse transcription (RT–qPCR)
MRTX1133 formic purchased from MedChemExpress. Usage Cited in: Nature. 2026 Jan;649(8098):1022-1031. [Abstract]
MRTX1133 (1, 10, 100 nM; 24 h) was used to treat KPC cells for 24 h, and the cells were analysed using immunoblotting.
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Cancer Cell
A pan-KRAS inhibitor and its derived degrader elicit multifaceted anti-tumor efficacy in KRAS-driven cancers. [Abstract]2025 Jul 25:S1535-6108(25)00310-1. PMID: 40780213 -
Immunity
Targeting intracellular oncoproteins with dimeric IgA promotes expulsion from the cytoplasm and immune-mediated control of epithelial cancers. [Abstract]2023 Nov 14;56(11):2570-2583.e6. PMID: 37909039
MRTX1133 formic purchased from MedChemExpress. Usage Cited in: Immunity. 2023 Nov 14;56(11):2570-2583.e6. [Abstract]
Western blot showing phosphorylated and total ERK1/2 in lysates of KRASG12D-mutated A427 cells, treated with anti-KRASG12D-IgA1 or MRTX1133.
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Cancer Discov
A plastic EMP1⁺ to LGR5⁺ cell state conversion as a bypass to KRAS-G12D pharmacological inhibition in metastatic colorectal cancer. [Abstract]2025 Oct 21. PMID: 41128661 -
Cancer Discov
Epigenetic and oncogenic inhibitors cooperatively drive differentiation and kill KRAS-mutant colorectal cancers. [Abstract]2024 Dec 2;14(12):2430-2449. PMID: 39121480 -
Cancer Discov
Oncogenic KRAS-dependent stromal interleukin-33 directs the pancreatic microenvironment to promote tumor growth. [Abstract]2024 Oct 4;14(10):1964-1989. PMID: 38958646
MRTX1133 formic purchased from MedChemExpress. Usage Cited in: Cancer Discov. 2024 Oct 4;14(10):1964-1989. [Abstract]
MRTX1133 (0.5 μM; 24 h)-treated 9805 cells produce conditioned medium (CM) that induces a reduction in Il33 expression in CD1WT fibroblasts.
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Nat Cancer
The MEK-RAF molecular glue IK-595 has potent antitumor activity across RAS/MAPK pathway-altered cancers. [Abstract]2026 Jan;7(1):116-130. PMID: 41482524 -
Cancer Res
Prolonged KRAS-MAPK Inhibition Induces Interferon Signaling That Promotes Cell State Transition and Confers Therapeutic Vulnerabilities. [Abstract]2026 Jul 2;86(13):3270-3286. PMID: 42008116 -
Cancer Res
KRAS Signaling Inhibition Induces a Targetable Metabolic Dependency on Lipophagy-Dependent Fatty Acid Oxidation in Pancreatic Cancer. [Abstract]2026 Apr 29:10.1158/0008-5472.CAN-24-1984. PMID: 42054558 -
Cancer Res
Targeted Degradation of SOS1 Exhibits Potent Anticancer Activity and Overcomes Resistance in KRAS-Mutant Tumors and BCR-ABL-Positive Leukemia. [Abstract]2025 Jan 2;85(1):101-117. PMID: 39437162 -
Cancer Res
Targeting BCL2 with Venetoclax Enhances the Efficacy of the KRASG12D Inhibitor MRTX1133 in Pancreatic Cancer. [Abstract]2024 Nov 4;84(21):3629-3639. PMID: 39137400
MRTX1133 formic purchased from MedChemExpress. Usage Cited in: Cancer Res. 2024 Nov 4;84(21):3629-3639. [Abstract]
PDAC cell lines grown in low confluent collagen cultures were treated with MRTX1133 (0.5 μmol/L) for 8 hours, and the effect on ERK1/2 phosphorylation was analyzed by Western blotting.
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Nat Commun
Oncogenic KRAS-driven type I interferon signalling primes pancreatic cancer for necroptosis. [Abstract]2026 Jun 15;17(1):5288. PMID: 42297776 -
Nat Commun
B7-H3-mediated cis-inhibition of EGFR by a tumor-selective bispecific antibody enhances anti-tumor efficacy and minimizes toxicities. [Abstract]2026 Feb 24;17(1):3113. PMID: 41735305 -
Nat Commun
2025 Dec 15. PMID: 41397976 -
Nat Commun
A decision point between transdifferentiation and programmed cell death priming controls KRAS-dependent pancreatic cancer development. [Abstract]2025 Feb 19;16(1):1765. PMID: 39971907 -
J Adv Res
Targeting class I HDACs suppresses oncogenic vulnerabilities and potentiates KRAS/MAPK pathway inhibitors in KRAS-mutant cancers. [Abstract]2026 Feb 13:S2090-1232(26)00144-X. PMID: 41692243 -
Adv Sci (Weinh)
Real-Time Visualization of Isoform-Specific RAF-KRAS Interactions in Living Cells Using FRET-BRET Hybrid Biosensors. [Abstract]2026 Feb 12:e15654. PMID: 41674298 -
Cell Rep Med
Syndecan-1-targeted therapeutic antibody impairs macropinocytosis and elicits antitumor immunity in pancreatic cancer. [Abstract]2026 Feb 17;7(2):102613. PMID: 41707651 -
Cell Rep Med
Developing a therapeutic elastase that stimulates anti-tumor immunity by selectively killing cancer cells. [Abstract]2025 Nov 18;6(11):102446. PMID: 41205593 -
Cell Rep Med
KRASG12D-driven pentose phosphate pathway remodeling imparts a targetable vulnerability synergizing with MRTX1133 for durable remissions in PDAC. [Abstract]2025 Feb 18;6(2):101966. PMID: 39970873 -
Sci Adv
2024 Dec 13;10(50):eadq4274. PMID: 39661665 -
Cell Death Dis
Simultaneous targeting of KRAS and CDK4 synergistically induces durable growth arrest in pancreatic cancer cells. [Abstract]2025 Dec 23. PMID: 41436723 -
Cancer Lett
Site-specific mutagenesis screening in KRASG12D mutant library to uncover resistance mechanisms to KRASG12D inhibitors. [Abstract]2024 Jun 1:591:216904. PMID: 38642608 -
Clin Cancer Res
Overcoming Adaptive Resistance to KRASG12D Blockade in Pancreatic Cancer through Vertical Pathway Inhibition. [Abstract]2026 Mar 9. PMID: 41801133 -
Clin Cancer Res
2024 Sep 13;30(18):4082-4099. PMID: 39018564 -
Acta Pharmacol Sin
143D, a novel selective KRASG12C inhibitor exhibits potent antitumor activity in preclinical models. [Abstract]2023 Jul;44(7):1475-1486. PMID: 36725884 -
Proc Natl Acad Sci U S A
Fra-2 controls the response to the KRAS inhibitor MRTX-1133 in pancreatic ductal adenocarcinoma. [Abstract]2026 May 5;123(18):e2601788123. PMID: 42054368 -
Proc Natl Acad Sci U S A
Genome-wide CRISPR screens in spheroid culture reveal that the tumor suppressor LKB1 inhibits growth via the PIKFYVE lipid kinase. [Abstract]2024 May 21;121(21):e2403685121. PMID: 38743625 -
Mol Med
MGST1 facilitates novel KRASG12D inhibitor resistance in KRASG12D-mutated pancreatic ductal adenocarcinoma by inhibiting ferroptosis. [Abstract]2024 Nov 5;30(1):199. PMID: 39501138 -
EMBO Mol Med
2025 Jun;17(6):1355-1392. PMID: 40329096 -
Cell Rep
Mutant KRAS-driven selective mRNA translation reveals mechanisms and therapeutic vulnerabilities in cancer. [Abstract]2026 Jun 23;45(6):117520. PMID: 42284146 -
Cell Rep
Accumulated sotorasib-bound KRASG12C reactivates the MAPK pathway and drives sotorasib resistance via the DHX9-RAC1-PAK1 axis. [Abstract]2026 May 4;45(5):117338. PMID: 42090290 -
Cell Rep
2024 Aug 22;43(9):114667. PMID: 39178114 -
Mol Cancer Ther
Dual Inhibitors of KRASG12D and HSP90 are Effective Against KRASG12D Inhibitor Resistance. [Abstract]2025 Oct 23. PMID: 41129140 -
ACS Appl Bio Mater
Differential Effects of Confinement on the Dynamics of Normal and Tumor-Derived Pancreatic Ductal Organoids. [Abstract]2024 Dec 16;7(12):8489-8502. PMID: 39576883 -
Sci Rep
Antimetastatic effects of MRTX1133 KRAS G12D specific inhibitor in a liver metastatic model of pancreatic ductal adenocarcinoma. [Abstract]2026 Jan 7;16(1):4144. PMID: 41501279 -
Transl Oncol
MRTX1133 attenuates KRASG12D mutated-colorectal cancer progression through activating ferroptosis activity via METTL14/LINC02159/FOXC2 axis. [Abstract]2024 Dec 9:52:102235. PMID: 39657309 -
Mol Oncol
KRASG 12C-inhibitor-based combination therapies for pancreatic cancer: insights from drug screening. [Abstract]2025 Feb;19(2):295-310. PMID: 39253995 -
ACS Pharmacol Transl Sci
Suite of Biochemical and Cell-Based Assays for the Characterization of Kirsten Rat Sarcoma (KRAS) Inhibitors and Degraders. [Abstract]2024 Dec 2;7(12):3921-3934. PMID: 39698278 -
STAR Protoc
Generating drug resistance models in human and murine cancer cell lines and assessing cross-resistance to chemotherapeutics and KRAS inhibitors. [Abstract]2026 Jun 26;7(3):104649. PMID: 42360881 -
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bioRxiv
The combination of BCL-xL PROTAC and mTOR inhibitor sensitizes pancreatic ductal adenocarcinoma to KRASG12D inhibitor treatment by enhancing apoptosis induction. [Abstract]2026 Jan 5:2026.01.05.697773. PMID: 41542622 -
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bioRxiv
VEGFR2 blockade overcomes acquired KRAS G12D inhibitor resistance driven by PI3Kγ activation. [Abstract]2025 Sep 18:2025.09.16.676456. PMID: 41000780 -
bioRxiv
Thioredoxin Reductase 1 inhibition triggers ferroptosis in KRAS-independent lung cancers. [Abstract]2025 Jul 30:2025.07.25.666783. PMID: 40766571 -
bioRxiv
Phosphoinositide 3-kinase regulates wild-type RAS signaling to confer resistance to KRAS inhibition. [Abstract]2025 Jun 25:2025.06.20.660715. PMID: 40666995 -
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bioRxiv
Resistance to the KRASG12D Inhibitor MRTX1133 is Associated with Increased Sensitivity to BET Inhibition. [Abstract]2025 May 13:2025.05.10.653074. PMID: 40463163 -
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bioRxiv
LKB1 suppresses growth and promotes the internalization of EGFR through the PIKFYVE lipid kinase. [Abstract]2023 Oct 20:2023.10.19.563158. PMID: 37904985 -
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Solvant et solubilité
DMSO : 50 mg/mL (77.32 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
H2O : 25 mg/mL (38.66 mM; ultrasonic and warming and heat to 60°C)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.08 mg/mL (3.22 mM); Clear solution
This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.08 mg/mL (3.22 mM); Clear solution
This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Working solution concentration: 0.22 mg/mL
This product has good water solubility, please refer to the measured solubility data in water/PBS/Saline for details.
Pureté et documentation
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Fiche technique (276 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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Instruction de manipulation (2659 KB)
Références
[1]. Wang X, et al. Identification of MRTX1133, a Noncovalent, Potent, and Selective KRASG12D Inhibitor [published online ahead of print, 2021 Dec 10]. J Med Chem. 2021;10.1021/acs.jmedchem.1c01688. [Content Brief]
[2]. KRAS G12D Inhibitor: MRTX1133. [(accessed on 22 April 2021)];2021 Available online.
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| H2O / DMSO | 1 mM | 1.5464 mL | 7.7320 mL | 15.4641 mL | 38.6602 mL |
| 5 mM | 0.3093 mL | 1.5464 mL | 3.0928 mL | 7.7320 mL | |
| 10 mM | 0.1546 mL | 0.7732 mL | 1.5464 mL | 3.8660 mL | |
| 15 mM | 0.1031 mL | 0.5155 mL | 1.0309 mL | 2.5773 mL | |
| 20 mM | 0.0773 mL | 0.3866 mL | 0.7732 mL | 1.9330 mL | |
| 25 mM | 0.0619 mL | 0.3093 mL | 0.6186 mL | 1.5464 mL | |
| 30 mM | 0.0515 mL | 0.2577 mL | 0.5155 mL | 1.2887 mL | |
| DMSO | 40 mM | 0.0387 mL | 0.1933 mL | 0.3866 mL | 0.9665 mL |
| 50 mM | 0.0309 mL | 0.1546 mL | 0.3093 mL | 0.7732 mL | |
| 60 mM | 0.0258 mL | 0.1289 mL | 0.2577 mL | 0.6443 mL |
* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.