| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Mutant BRAF (including V600E, G469A, G466V, and the p61-BRAFV600E splice variant). CFT1946 is a bifunctional degradation activating compound (BiDACTM) degrader comprising a BRAF kinase domain targeting ligand linked to a cereblon ligand. It selectively degrades mutant BRAF with a DC50 of 14 nmol/L for BRAF V600E (at 24 hours). [1,2]
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| ln Vitro |
BRAF V600E Degradation and Pathway Inhibition: In A375 cells (BRAF V600E mutant), CFT1946 degrades BRAF V600E with a DC50 of 14 nmol/L and an Emax of 26% at 24 hours. It inhibits ERK phosphorylation with an IC50 of 11 nmol/L at 24 hours and inhibits cell proliferation with a GI50 of 94 nmol/L at 96 hours. [1,2]
Mechanism of Degradation Validation: CFT1946-induced BRAF V600E degradation is dependent on cereblon and the proteasome. Degradation is blocked by competition with a cereblon ligand, the CUL4 E3 ligase inhibitor MLN4924, or the proteasome inhibitor bortezomib. [2] Activity Against Non-V600 Mutants: CFT1946 degrades BRAF G469A (Class II), G466V (Class III) mutants, and the p61-BRAFV600E splice variant (as shown in heterologous expression systems in HEK293T cells). In the BRAF G466V heterozygous lung tumor cell line H1666, CFT1946 inhibits proliferation, whereas encorafenib is ineffective. [1,2] Selectivity: CFT1946 exhibits exquisite selectivity for BRAF V600E. Proteome profiling in A375 cells shows degradation selectivity is limited to BRAF V600E, with no effect on wild-type BRAF or CRAF. In HepG2 cells (wild-type BRAF), the GI50 is >10 μmol/L, and there is no activity in KRAS-mutant HCT116 cells. [1,2] Activity in Resistance Models: In an engineered BRAF inhibitor resistance model, A375-BRAFV600E/NRASQ61K double mutant cells, CFT1946 effectively degrades BRAF V600E, with an IC50 of 42 nmol/L for pERK inhibition (at 1 hour) and a GI50 of 150 nmol/L for cell proliferation inhibition (at 96 hours). [2] |
| ln Vivo |
Antitumor Efficacy: In A375 xenograft models (BRAF V600E), oral administration of CFT1946 (10 mg/kg, BID) results in sustained tumor regression, identified as the minimum efficacious dose. In the BRAFi-resistant A375-NRASQ61K xenograft model, CFT1946 as a single agent and in combination with a MEK inhibitor shows tumor growth inhibition and regression, demonstrating superior efficacy compared to a BRAF inhibitor. [2]
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| Enzyme Assay |
BRAF Kinase Inhibition Constant: The inhibitory constant (Ki) of CFT1946 for BRAF V600E was determined via kinase activity assays to be 0.1 nmol/L. [2]
Cereblon Binding Affinity: The dissociation constant (Kd) of CFT1946 for cereblon was determined using fluorescence polarization to be 11 nmol/L. [2] |
| Cell Assay |
HiBiT Degradation Assay: A cell line expressing HiBiT-tagged BRAF V600E was treated with various concentrations of CFT1946. After lysis, luminescence was measured to quantify protein levels for DC50 and Emax calculations. [2]
Cell Proliferation Inhibition Assay: Cell lines such as A375, HCT116, and H1666 were seeded in plates and treated with various concentrations of CFT1946 for 96 hours. Cell viability was assessed to calculate GI50 values. [1,2] Western Blot Analysis: Cells treated with CFT1946 were lysed, and protein levels of BRAF V600E, pERK, and total ERK were detected using specific antibodies to assess target degradation and downstream signaling inhibition. [2] Mechanism of Action Validation: A375 cells were treated with CFT1946 alone or in combination with a cereblon ligand, MLN4924, or bortezomib. BRAF V600E levels were analyzed by Western blot to confirm mechanism dependency. [2] |
| Animal Protocol |
In Vivo Efficacy Study (A375 Xenograft): A375 cells were implanted subcutaneously in mice. When tumors reached a certain size, mice were randomized and treated orally with various doses of CFT1946 (including 10 mg/kg) twice daily. Tumor volumes were measured to assess antitumor activity and determine the minimum efficacious dose. [2]
In Vivo Efficacy Study (Resistance Model): A375-NRASQ61K cells were implanted subcutaneously in mice. When tumors reached a certain size, mice were randomized and treated orally with CFT1946 alone or in combination with a MEK inhibitor twice daily. Tumor volumes were measured to evaluate efficacy in a resistance model. [2] In Vivo Pharmacokinetic Study: In mice and rats, CFT1946 was administered intravenously or orally. Blood samples were collected, and plasma drug concentrations were analyzed by LC-MS/MS to calculate pharmacokinetic parameters. [2] |
| ADME/Pharmacokinetics |
Mouse Pharmacokinetics: CFT1946 exhibits an observed clearance (CLobs) of 0.8 mL/min/kg and an oral bioavailability (F%) of 89% in mice. [2]
Rat Pharmacokinetics: CFT1946 exhibits an observed clearance (CLobs) of 0.5 mL/min/kg and an oral bioavailability (F%) of 89% in rats. [2] |
| Toxicity/Toxicokinetics |
Based on the provided preclinical data (AACR 2022 abstract and C4 Therapeutics presentation), no specific toxicity or toxicokinetic parameters (such as maximum tolerated dose, organ-specific toxicity, or plasma protein binding) were detailed for Tagarafdeg (CFT1946). The available information focuses on its favorable pharmacokinetic profile and tolerability as inferred from its advancement into clinical trials. Key indicators include its high oral bioavailability (89% in both mice and rats), low in vivo clearance (0.8 mL/min/kg in mice, 0.5 mL/min/kg in rats), and a preclinical profile described as "balanced," which supported its progression into Phase 1 studies for patients with BRAF V600X-driven cancers, including those resistant to prior BRAF inhibitor therapy.
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| References |
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| Additional Infomation |
Background and Mechanism of Action: BRAF kinase is a critical node in the MAPK signaling pathway, mutated in approximately 8% of human cancers. Currently approved BRAF inhibitors (e.g., vemurafenib, dabrafenib, encorafenib) are effective against V600 mutations but resistance typically emerges within a year, and they are ineffective against non-V600 BRAF mutants (Class II and III). CFT1946 is a bifunctional degradation activating compound (BiDACTM) that recruits cereblon to BRAF, inducing ubiquitination and proteasomal degradation of mutant BRAF proteins (including V600E, G469A, G466V, and splice variants). This degradation strategy avoids paradoxical activation of wild-type RAF and may overcome multiple resistance mechanisms. [1,2]
Clinical Development: Based on its promising preclinical activity, selectivity, and pharmacokinetic profile, CFT1946 has advanced into Phase 1 clinical trials for patients with BRAF V600X-driven cancers and those with inhibitor-resistant BRAF V600X-driven cancers. [2] |
| Molecular Formula |
C45H49F2N11O9S
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|---|---|
| Molecular Weight |
958.000674962997
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| Exact Mass |
957.34
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| Elemental Analysis |
C, 56.42; H, 5.16; F, 3.97; N, 16.08; O, 15.03; S, 3.35
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| CAS # |
2882165-79-7
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| PubChem CID |
166168413
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| Appearance |
Off-white to light yellow solid powder
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| LogP |
2.7
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
17
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| Rotatable Bond Count |
11
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| Heavy Atom Count |
68
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| Complexity |
2110
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CN1N=C(N2CCC(NC2=O)=O)C2=CC(=C(N3CCC(CC3)(O)CC(N3CCC4(OC[C@H](N5C=NC6=CC=C(C=C6C5=O)OC5C(=CC=C(C=5C#N)NS(=O)(=O)N(C)CC)F)C4)CC3)=O)C=C21)F
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| InChi Key |
OCDRMYDQTIPVOI-HHHXNRCGSA-N
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| InChi Code |
InChI=1S/C45H49F2N11O9S/c1-4-53(2)68(64,65)52-35-8-6-32(46)40(31(35)24-48)67-28-5-7-34-29(19-28)42(61)58(26-49-34)27-22-45(66-25-27)12-17-56(18-13-45)39(60)23-44(63)10-15-55(16-11-44)37-21-36-30(20-33(37)47)41(51-54(36)3)57-14-9-38(59)50-43(57)62/h5-8,19-21,26-27,52,63H,4,9-18,22-23,25H2,1-3H3,(H,50,59,62)/t27-/m1/s1
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| Chemical Name |
(3R)-3-[6-[2-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluorophenoxy]-4-oxoquinazolin-3-yl]-8-[2-[1-[3-(2,4-dioxo-1,3-diazinan-1-yl)-5-fluoro-1-methylindazol-6-yl]-4-hydroxypiperidin-4-yl]acetyl]-1-oxa-8-azaspiro[4.5]decane
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| Synonyms |
CFT1946; CFT 1946; 2882165-79-7; CFT-1946; N'-[2-Cyano-4-fluoro-3-[[3-[(3R)-8-[2-[1-[5-fluoro-1-methyl-3-(tetrahydro-2,4-dioxo-1(2H)-pyrimidinyl)-1H-indazol-6-yl]-4-hydroxy-4-piperidinyl]acetyl]-1-oxa-8-azaspiro[4.5]dec-3-yl]-3,4-dihydro-4-oxo-6-quinazolinyl]oxy]phenyl]-N-ethyl-N-methylsulfamide;
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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 Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| 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 :~100 mg/mL (~104.38 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 | 1.0438 mL | 5.2192 mL | 10.4384 mL | |
| 5 mM | 0.2088 mL | 1.0438 mL | 2.0877 mL | |
| 10 mM | 0.1044 mL | 0.5219 mL | 1.0438 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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