| Size | Price | Stock | Qty |
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| 1mg |
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| Targets |
The compound targets the active GTP-bound state of the KRAS G12D mutant, binding to the switch-II (S-II) pocket. KD-8 is a covalent inhibitor, and its ability to suppress KRAS G12D activity is demonstrated by decreased levels of active KRAS-GTP in KRAS G12D-mutant cancer cells, with selectivity for G12D over G13D and wild-type KRAS. By binding to KRAS G12D, it prevents downstream signaling through the RAF-MEK-ERK pathway.
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| ln Vitro |
Against three KRAS G12D mutant cells (Panc1, SW1990, and CT26), KRAS G12D inhibitor 14 (compound KD-8) demonstrated antiproliferative action with an IC50 of 2.1 μM. In KRAS G12D mutant cancer cell lines (CT26 and SW1990), KRAS G12D inhibitor 14 decreases the active form of KRAS (KRAS-GTP); however, this effect is not shown in KRAS G13D mutant cancer cell lines (HCT116). Moreover, KD-8 effectively triggered apoptosis in the cancer cells CT26 and SW1990. In CT26 and SW1990 cancer cell lines, KRAS G12D inhibitor 14 downregulates phosphorylated Raf and Erk, but not in HeLa cells (KRAS WT) [1].
In biochemical assays, KD-8 binds to the KRAS G12D protein with a binding affinity (Kd) of 33 nM, as determined by isothermal titration calorimetry (ITC). The compound shows high anti-proliferative activity against three KRAS G12D-mutated cancer cell lines (Panc1 pancreatic, SW1990 pancreatic, and CT26 colorectal) with an average IC50 of 2.1 uM. It reduces the levels of active KRAS-GTP in KRAS G12D-mutant cell lines and downregulates phospho-Raf and phospho-Erk in a dose-dependent manner, with no such effects observed in KRAS G13D-mutant or wild-type KRAS cells. |
| ln Vivo |
KRAS G12D inhibitor 14 (compound KD-8) (40 mg/kg or 60 mg/kg; i.p.) displayed significant anticancer activity in the CT26 tumor model, with a tumor growth inhibition (TGI) of 42% or 53 %, and will not cause significant toxicity [1].
In a CT26 syngeneic mouse tumor model, KD-8 (40 mg/kg or 60 mg/kg; intraperitoneal administration) displayed significant anticancer activity, with tumor growth inhibition (TGI) values of 42% and 53%, respectively. The compound did not cause significant toxicity at the tested doses, as indicated by a lack of notable body weight loss or other observable adverse effects. These results confirm the in vivo efficacy of KD-8 in a KRAS G12D-driven tumor model. |
| Enzyme Assay |
The binding affinity between KD-8 and KRAS G12D protein is determined using isothermal titration calorimetry (ITC). Purified recombinant KRAS G12D protein is placed in the sample cell, and a concentrated solution of KD-8 is titrated in. The heat change accompanying each injection is measured, and the resulting binding isotherm is fitted to calculate the binding affinity (Kd) and thermodynamic parameters (deltaH, deltaS). The binding mode to the switch-II pocket is confirmed by X-ray crystallography or molecular docking studies.
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| Cell Assay |
Cellular anti-proliferative activity is assessed in KRAS G12D-mutant cell lines (Panc1, SW1990, CT26) and control cells (KRAS G13D-mutant HCT116 or KRAS wild-type cells). Cells are seeded in 96-well plates and treated with varying concentrations of KD-8 (0.1-20 uM) for 72-96 hours. Cell viability is measured using MTT, CCK-8, or CellTiter-Glo assays to calculate IC50 values. To assess pathway inhibition, cells are treated with KD-8 (0.5-10 uM) for 4-24 hours, lysed, and analyzed by Western blot using antibodies against KRAS, phospho-Raf (Ser338), phospho-ERK (Thr202/Tyr204), total ERK, and KRAS-GTP pull-down assays. Apoptosis is assessed by measuring cleaved caspase-3 and PARP by Western blot.
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| Animal Protocol |
In a CT26 syngeneic mouse tumor model, female BALB/c mice (6-8 weeks old) are injected subcutaneously with CT26 cells. When tumors reach an average volume of approximately 100-150 mm3, mice are randomized into treatment groups. KD-8 is formulated in a suitable vehicle (e.g., 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline) and administered via intraperitoneal (i.p.) injection at doses of 40 mg/kg or 60 mg/kg, typically once daily for 2-3 weeks. Tumor volume is measured by calipers twice weekly, and body weight is monitored as a toxicity indicator. At study termination, tumors are harvested for analysis of KRAS-GTP levels and phospho-ERK by Western blot and for histopathological examination.
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| ADME/Pharmacokinetics |
Specific pharmacokinetic parameters for KD-8 are not detailed. The compound is administered via intraperitoneal injection in mouse efficacy studies (40-60 mg/kg). Key PK properties such as bioavailability, half-life, and plasma/tissue exposure would require empirical determination. The compound's ability to achieve sufficient tumor concentrations to inhibit KRAS G12D is supported by its observed in vivo efficacy.
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| Toxicity/Toxicokinetics |
KD-8 is reported not to cause significant toxicity in CT26 tumor-bearing mice at doses of 40-60 mg/kg, as assessed by body weight monitoring and general observations. No specific hematological or histopathological toxicological data are detailed. However, the compound is specific for KRAS G12D, and normal cells expressing wild-type KRAS are not targeted, suggesting a potentially favorable safety window. Full toxicological evaluation would be required for clinical development.
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| References | |
| Additional Infomation |
KD-8 is a research-grade chemical tool for studying KRAS G12D-driven cancers and validating KRAS G12D as a therapeutic target. The KRAS G12D mutation is one of the most common KRAS mutations in human cancers, present in approximately 50% of pancreatic adenocarcinomas, 30-40% of colorectal cancers, and 15-20% of non-small cell lung cancers. KD-8 is a covalent inhibitor that binds to the GTP-ON state of KRAS G12D, representing a distinct pharmacological class from non-covalent inhibitors. As of the latest updates, KD-8 has not been approved for clinical use.
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| Molecular Formula |
C20H19F3N4OS
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|---|---|
| Molecular Weight |
420.451273202896
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| Exact Mass |
420.123
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| CAS # |
2765254-39-3
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| PubChem CID |
163196413
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| Appearance |
Off-white to light yellow solid powder
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| LogP |
4.5
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
29
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| Complexity |
597
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C12CCC(NC1)CN2C1N=CN=C2SC(C3=CC(C(F)(F)F)=CC=C3OC)=CC2=1
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| InChi Key |
DYYANULKLUNJSO-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C20H19F3N4OS/c1-28-16-5-2-11(20(21,22)23)6-14(16)17-7-15-18(25-10-26-19(15)29-17)27-9-12-3-4-13(27)8-24-12/h2,5-7,10,12-13,24H,3-4,8-9H2,1H3
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| Chemical Name |
4-(2,5-diazabicyclo[2.2.2]octan-2-yl)-6-[2-methoxy-5-(trifluoromethyl)phenyl]thieno[2,3-d]pyrimidine
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
DMSO : ~50 mg/mL (~118.92 mM)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3784 mL | 11.8920 mL | 23.7840 mL | |
| 5 mM | 0.4757 mL | 2.3784 mL | 4.7568 mL | |
| 10 mM | 0.2378 mL | 1.1892 mL | 2.3784 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.
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.
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