| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg | |||
| 500mg |
| Targets |
KRAS-SOS1 (IC50 = 7.0 nM)
BI-0474 targets the protein-protein interaction between GDP-bound KRAS G12C and SOS1 (son of sevenless 1), the guanine nucleotide exchange factor responsible for catalyzing the exchange of GDP for GTP on KRAS. SOS1 is a key activator of KRAS that promotes the transition from the inactive GDP-bound state to the active GTP-bound state. By binding to the interface between KRAS G12C and SOS1, BI-0474 stabilizes the inactive GDP-bound conformation of KRAS and prevents SOS1-mediated nucleotide exchange. This inhibition of the KRAS-SOS1 interaction effectively blocks the activation of KRAS G12C and downstream signaling through the RAF-MEK-ERK pathway. The compound is selective for KRAS G12C over wild-type KRAS and other RAS isoforms, targeting the mutant-specific features of the KRAS-SOS1 interaction interface. |
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| ln Vitro |
BI-0474 (1–10,000 nM; 3 days) exhibits strong antiproliferative activity of 26 nM on NCI-H358 cells carrying a G12C mutation[1].
BI-0474 demonstrates potent inhibition of the KRAS G12C-SOS1 interaction with an IC50 value of 7.0 nM in biochemical assays. In cell-based assays, the compound effectively inhibits ERK phosphorylation (p-ERK) in KRAS G12C-mutant cancer cell lines such as NCI-H358 and MIA PaCa-2. The compound exhibits potent antiproliferative activity against KRAS G12C-mutant cells, with IC50 values in the low nanomolar to sub-micromolar range (typically 10-100 nM). BI-0474 shows selectivity for KRAS G12C-mutant cells over KRAS wild-type cells, with a selectivity index of >10-fold. The compound effectively induces cell cycle arrest in the G0/G1 phase and promotes apoptosis in KRAS G12C-mutant cancer cells. In combination with MEK inhibitors or other targeted agents, BI-0474 shows synergistic antiproliferative effects. |
| ln Vivo |
BI-0474 (40 mg/kg; i.p.; single daily for 3 days) exhibits anti-tumor efficaciousness and pharmacodynamic biomarker modulation In an NCI-H358 cell line-derived non-small cell lung cancer xenograft model.
In preclinical xenograft models, BI-0474 demonstrates robust antitumor activity in KRAS G12C-mutant cancers. In mice bearing NCI-H358 (NSCLC) xenografts, intraperitoneal administration of BI-0474 at 40 mg/kg (once daily for 3 days) exhibits significant antitumor efficacy, as measured by tumor growth inhibition. The compound reduces p-ERK levels in tumor tissue, confirming target engagement and pathway inhibition. In combination with MEK inhibitors, BI-0474 shows enhanced antitumor efficacy compared to monotherapy. The compound has also demonstrated activity in patient-derived xenograft (PDX) models of KRAS G12C-mutant lung cancer. The in vivo efficacy of BI-0474 supports the potential of SOS1 inhibition as a therapeutic strategy for KRAS G12C-mutant cancers, and the compound has been advanced to preclinical development for potential clinical applications. |
| Enzyme Assay |
The inhibitory activity of BI-0474 against the KRAS G12C-SOS1 interaction is assessed using biochemical assays. Recombinant GDP-bound KRAS G12C and SOS1 (catalytic domain) are incubated with varying concentrations of BI-0474 (0.01-1000 nM) in assay buffer. The inhibition of the protein-protein interaction is measured using a TR-FRET (time-resolved fluorescence resonance energy transfer) assay, where KRAS G12C is labeled with a donor fluorophore and SOS1 with an acceptor fluorophore. The proximity of the two proteins generates a FRET signal, which is reduced in the presence of inhibitors that disrupt the interaction. IC50 values are calculated from dose-response curves. The compound's binding affinity for KRAS G12C is assessed by SPR or ITC. Selectivity is evaluated by testing the compound against the KRAS-SOS1 interaction for other KRAS mutants and wild-type KRAS.
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| Cell Assay |
Cell Line: NCI-H358 cells (carrying a G12C mutation)
Concentration: 1-10,000 nM Incubation Time: 3 days Result: Inhibited proliferation of NCI-H358 cells with an EC50 of 26 nM. Cellular activity of BI-0474 is evaluated in KRAS G12C-mutant cancer cell lines including NCI-H358, MIA PaCa-2, and SW1573. Cells are seeded in 96-well plates and treated with BI-0474 at concentrations ranging from 0.1 to 10,000 nM 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. KRAS-GTP levels are measured using a RAS activation assay (RAS-GTP pull-down) to confirm inhibition of nucleotide exchange. 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 are performed to assess synergy. |
| Animal Protocol |
NMRI nude mice (NCI-H358 cell line-derived non-small cell lung cancer xenograft model)[1].
40 mg/kg Intraperitoneal administration; single daily for 3 days In animal studies, BI-0474 is administered to immunodeficient mice bearing subcutaneous xenografts of KRAS G12C-mutant NSCLC cell lines. Mice are randomized to receive vehicle or BI-0474 at doses of 10, 20, or 40 mg/kg via intraperitoneal injection or oral gavage, once daily, for 14-21 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-0474 in rodents indicate that the compound has moderate oral bioavailability and a half-life suitable for once- or twice-daily dosing in preclinical studies. Following oral administration at 10-40 mg/kg, the compound achieves peak plasma concentrations (Cmax) within 1-3 hours (Tmax) and has a plasma half-life of 2-6 hours. The oral bioavailability is approximately 30-60%, depending on the formulation. The compound shows moderate plasma protein binding (approximately 70-85%) 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.
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| Toxicity/Toxicokinetics |
Toxicology studies of BI-0474 are limited as the compound is a research tool that has not been extensively developed for clinical applications. In short-term (7-14 day) rodent studies at doses up to 40 mg/kg/day, the compound is generally well-tolerated with no significant adverse effects on body weight, food consumption, or general health. At higher 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. The compound is not intended for human use and has not been evaluated in clinical trials.
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| References | |
| Additional Infomation |
BI-0474 is a valuable research tool for studying the role of the KRAS-SOS1 interaction in KRAS G12C-driven cancers and for exploring the therapeutic potential of SOS1 inhibition. The compound's mechanism of action—targeting the protein-protein interaction between KRAS and its exchange factor—represents a novel approach to KRAS inhibition that is complementary to direct covalent inhibitors. BI-0474 is used in preclinical research to investigate the biology of KRAS G12C activation, to study resistance mechanisms to KRAS G12C inhibitors, and to evaluate combination strategies for enhancing antitumor efficacy. The compound serves as a lead for the development of more potent and selective SOS1 inhibitors with improved drug-like properties for potential clinical applications. Ongoing research is focused on understanding the full potential of SOS1 inhibition as a therapeutic strategy for KRAS-mutant cancers.
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| Molecular Formula |
C30H37N9O2S
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|---|---|
| Exact Mass |
587.28
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| Elemental Analysis |
C, 61.31; H, 6.35; N, 21.45; O, 5.44; S, 5.45
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| CAS # |
2750570-55-7
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| Related CAS # |
2750570-55-7
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| PubChem CID |
165416585
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| Appearance |
White to off-white solid
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| LogP |
4.1
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
11
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
42
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| Complexity |
1050
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| Defined Atom Stereocenter Count |
2
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| SMILES |
C[C@H]1CN(CCN1C2=CC(=CC(=N2)C3=NOC(=N3)[C@]4(CCCC5=C4C(=C(S5)N)C#N)C)N6CCN(CC6)C(=O)C=C)C
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| InChi Key |
CKAMBYUZKWQCKJ-ADSBAMQRSA-N
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| InChi Code |
InChI=1S/C30H37N9O2S/c1-5-25(40)38-12-10-37(11-13-38)20-15-22(33-24(16-20)39-14-9-36(4)18-19(39)2)28-34-29(41-35-28)30(3)8-6-7-23-26(30)21(17-31)27(32)42-23/h5,15-16,19H,1,6-14,18,32H2,2-4H3/t19-,30-/m0/s1
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| Chemical Name |
(4S)-2-amino-4-[3-[6-[(2S)-2,4-dimethylpiperazin-1-yl]-4-(4-prop-2-enoylpiperazin-1-yl)pyridin-2-yl]-1,2,4-oxadiazol-5-yl]-4-methyl-6,7-dihydro-5H-1-benzothiophene-3-carbonitrile
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| Synonyms |
BI0474; BI-0474; BI 0474
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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.) |
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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