| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg | |||
| 500mg |
| Targets |
BI-2865 targets KRAS, a small GTPase that functions as a molecular switch in cell signaling. KRAS is the most frequently mutated oncogene in human cancers, with mutations occurring in approximately 25% of all tumors. Common activating mutations include G12C, G12D, G12V, and G13D, which impair the intrinsic GTPase activity of KRAS and its interaction with GTPase-activating proteins (GAPs), leading to accumulation of the active GTP-bound form and constitutive activation of downstream signaling. BI-2865 binds to the switch II region of KRAS in the GDP-bound state, stabilizing the inactive conformation and preventing nucleotide exchange and effector binding. Unlike covalent G12C inhibitors that require the mutant cysteine for binding, BI-2865 binds through non-covalent interactions, allowing it to target a broader range of KRAS mutants.
|
|---|---|
| ln Vitro |
BI-2865 is a port aminoalcohol substituent and terminal pin connection equivalent that is labeled. E62 and the R68 side chain of BI-2865 (5 days) have a direct ionic link with BI-2865, as demonstrated by the cocrystal structure of BI-2865 bound to KRAS. G12C, G12D, or G12V expression is inhibited when IL-13 is present. Water-mediated hydrogen bonding network formed by average IC50 and Q61 backbone locations in BaF3 cells with mutant KRAS during their growth. In the neighborhood of 140 nM[1].
BI-2865 demonstrates potent binding affinity for multiple KRAS variants. The compound binds to KRAS WT with a Kd of 6.9 nM, KRAS G12C with a Kd of 4.5 nM, KRAS G12D with a Kd of 32 nM, KRAS G12V with a Kd of 26 nM, and KRAS G13D with a Kd of 4.3 nM. In cell-based assays, BI-2865 inhibits the proliferation of BaF3 cells expressing KRAS G12C, G12D, or G12V mutants with average IC50 values of approximately 140 nM. The compound effectively reduces ERK phosphorylation (p-ERK) in KRAS-mutant cancer cell lines, confirming inhibition of downstream MAPK signaling. BI-2865 shows selectivity for KRAS over other RAS isoforms (HRAS, NRAS) and does not significantly inhibit a panel of other kinases at concentrations up to 10 μM. |
| ln Vivo |
In vivo, BI-2865 has demonstrated antitumor activity in preclinical models of KRAS-mutant cancers. In mouse xenograft models bearing KRAS G12C-mutant or G12D-mutant tumors, oral administration of BI-2865 at doses of 30-100 mg/kg (once or twice daily) results in significant tumor growth inhibition. The compound reduces p-ERK levels in tumor tissue, confirming target engagement and pathway modulation. In combination with MEK inhibitors or other targeted therapies, BI-2865 shows enhanced antitumor efficacy compared to monotherapy. The compound also demonstrates activity in models of acquired resistance to G12C-specific inhibitors, highlighting the potential of pan-KRAS inhibition to overcome resistance mechanisms. However, as a research tool, comprehensive in vivo efficacy data across multiple models is still emerging.
|
| Enzyme Assay |
The binding affinity of BI-2865 to KRAS is assessed using biochemical assays. Recombinant KRAS proteins (wild-type and mutants) are loaded with GDP and incubated with varying concentrations of BI-2865 (0.1-1000 nM). The binding affinity is measured by isothermal titration calorimetry (ITC) or surface plasmon resonance (SPR), with Kd values calculated from binding isotherms. The compound's ability to inhibit SOS-mediated nucleotide exchange (GDP to GTP) is assessed using a fluorescent nucleotide exchange assay with the exchange factor SOS. The inhibition of RAF binding is measured using a TR-FRET assay in which KRAS is incubated with RAF-RBD in the presence of varying concentrations of BI-2865. IC50 values for nucleotide exchange inhibition and RAF binding inhibition are calculated from dose-response curves.
|
| Cell Assay |
Cellular activity of BI-2865 is evaluated in KRAS-mutant cancer cell lines including NCI-H358 (G12C), MIA PaCa-2 (G12C), and PANC-1 (G12D). Cells are seeded in 96-well plates and treated with BI-2865 at concentrations ranging from 0.01 to 100 μM for 48-72 hours. Cell viability is assessed by CellTiter-Glo or MTT assays, and IC50 values are calculated. ERK phosphorylation (p-ERK) is measured by Western blot or AlphaLISA after 2-24 hours of treatment. Apoptosis is assessed by caspase-3/7 activity and annexin V/PI staining. Cell cycle analysis is performed by flow cytometry. Combination studies with MEK inhibitors or other agents are performed to assess synergy. Target engagement in cells is confirmed by assessing the displacement of fluorescently labeled KRAS-binding probes.
|
| Animal Protocol |
In animal studies, BI-2865 is administered to immunodeficient mice bearing subcutaneous xenografts of KRAS-mutant NSCLC or pancreatic cancer cell lines. Mice are randomized to receive vehicle or BI-2865 at doses of 10, 30, or 100 mg/kg via oral gavage, once or twice daily, for 14-28 days. Tumor volume is measured twice weekly with calipers, and tumor growth inhibition is calculated. Body weight is monitored to assess tolerability. At study termination, tumors are harvested for analysis of p-ERK levels (by Western blot or IHC), Ki67 (proliferation marker), and cleaved caspase-3 (apoptosis marker). Blood and plasma are collected for pharmacokinetic analysis. Pharmacodynamic biomarkers are measured in plasma or tumor tissue to confirm target engagement and pathway modulation.
|
| ADME/Pharmacokinetics |
Pharmacokinetic studies of BI-2865 in rodents indicate that the compound has moderate to good oral bioavailability and a half-life suitable for once- or twice-daily dosing in preclinical studies. Following oral administration at 10-30 mg/kg, the compound achieves peak plasma concentrations (Cmax) within 1-3 hours (Tmax) and has a plasma half-life of 3-8 hours. The oral bioavailability is approximately 40-70%, depending on the formulation. The compound shows moderate plasma protein binding (approximately 70-90%) and distributes to tissues including tumor tissue. Metabolism is primarily via CYP450 enzymes, and the compound is excreted in feces and urine. As a research compound, comprehensive PK studies are limited, and the compound is primarily used for proof-of-concept studies.
|
| Toxicity/Toxicokinetics |
Toxicology studies of BI-2865 are limited as the compound is a research tool rather than a clinical candidate. In short-term (7-14 day) rodent studies at doses up to 100 mg/kg/day, the compound is generally well-tolerated with no significant adverse effects on body weight, food consumption, or general health. At high doses, mild gastrointestinal effects and transient elevations in liver enzymes may occur. No significant hematological abnormalities or histopathological changes have been reported at therapeutic doses. As a pan-KRAS inhibitor, potential on-target toxicities related to inhibition of wild-type KRAS in normal tissues (e.g., gastrointestinal and skin effects) are a theoretical concern, though these have not been systematically evaluated for BI-2865. The compound is not intended for human use and has not been evaluated in clinical trials.
|
| References | |
| Additional Infomation |
BI-2865 is a breakthrough research tool that demonstrates the feasibility of non-covalent pan-KRAS inhibition, a concept that was long considered challenging due to the difficulty of targeting the relatively flat and featureless surface of KRAS. The compound's ability to bind with high affinity to multiple KRAS mutants, including G12D which is the most common KRAS mutation and lacks a cysteine for covalent targeting, represents a significant advancement in KRAS drug discovery. BI-2865 is widely used in preclinical research to study the biology of pan-KRAS inhibition, to investigate resistance mechanisms, and to evaluate combination strategies. The compound serves as a valuable reference for the development of clinical-stage pan-KRAS inhibitors and has contributed to the growing understanding of KRAS as a druggable target. Ongoing research is focused on optimizing the pharmacokinetic properties and selectivity profile of pan-KRAS inhibitors for clinical development.
|
| Molecular Formula |
C23H27N7O2S
|
|---|---|
| Molecular Weight |
465.571182489395
|
| Exact Mass |
465.19
|
| Elemental Analysis |
C, 59.34; H, 5.85; N, 21.06; O, 6.87; S, 6.89
|
| CAS # |
2937327-93-8
|
| Related CAS # |
2937327-93-8
|
| PubChem CID |
168268166
|
| Appearance |
Light yellow to brown solid
|
| Density |
1.38±0.1 g/cm3(Temp: 20 °C; Press: 760 Torr)(Predicted)
|
| Boiling Point |
708.7±70.0 °C(Predicted)
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
10
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
33
|
| Complexity |
752
|
| Defined Atom Stereocenter Count |
3
|
| SMILES |
C12CCC[C@](C)(C3ON=C(C4=NC=CC(O[C@H]([C@@H]5CCCN5C)C)=N4)N=3)C=1C(C#N)=C(N)S2
|
| InChi Key |
MIUFORKWYHBPRW-HMFCALDFSA-N
|
| InChi Code |
InChI=1S/C23H27N7O2S/c1-13(15-6-5-11-30(15)3)31-17-8-10-26-20(27-17)21-28-22(32-29-21)23(2)9-4-7-16-18(23)14(12-24)19(25)33-16/h8,10,13,15H,4-7,9,11,25H2,1-3H3/t13-,15-,23-/m0/s1
|
| Chemical Name |
(4S)-2-amino-4-methyl-4-[3-[4-[(1S)-1-[(2S)-1-methylpyrrolidin-2-yl]ethoxy]pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl]-6,7-dihydro-5H-1-benzothiophene-3-carbonitrile
|
| Synonyms |
BI2865; BI-2865; BI 2865
|
| 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 (In Vitro) |
DMSO: ~250 mg/mL (~537.0 mM)
|
|---|---|
| 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 | 2.1479 mL | 10.7395 mL | 21.4790 mL | |
| 5 mM | 0.4296 mL | 2.1479 mL | 4.2958 mL | |
| 10 mM | 0.2148 mL | 1.0740 mL | 2.1479 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.
|
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
