Table 3 Target therapies for CRAF/pan-RAF kinases
From: Targeting CRAF kinase in anti-cancer therapy: progress and opportunities
Compounds | Targets (IC50 or Kd values) | Administration efficiency | Clinical trial status and efficacy | Ref. | |
|---|---|---|---|---|---|
In vitro | In vivo usage | ||||
Selective CRAF inhibitors | |||||
 GW 5074 | 9 nM - CRAF | GW5074 potentiates the cytotoxicity of Sorafenib through mitochondrial dysfunction | GW5074 (25 mg/kg, IP) combination with Sorafenib in ACHN RCC tumors | NCT03406364, Phase I , Combined GW5074 and Sorafenib to treat solid tumor | |
 ZM 336372 | 0.07 μM - CRAF | ZM336372 suppresses carcinoid tumor cell proliferation and induces cell cycle inhibitors p21 and p18 | NO | NO | [25] |
 SHR902275 | 1.6 nM - CRAF 5.7 nM - BRAF V600E 10 nM - BRAF WT | SHR902275 shows cell growth inhibition with GI50 of 1.5 and 0.17 nM, 0.4 nM, and 0.32 nM for H358, A375, Calu6, and SK-MEL2 cells | SHR902275 (3-30 mg/kg, orally) inhibits cancer progression in RAS mutant Calu6 CDX model | NO | [26] |
 RAF inhibitor 2t | 50 nM - CRAF | 2t exhibited potent activities on WM3629 cell lines (IC50 0.56–0.86 μM) | NO | NO | [27] |
 RAF inhibitor 10c | 8.79 nM - CRAF 38.3 nM - BRAF V600E | 10c were (IC50 1.82 μM and 2.73 nM) against the A375P and U937 cell lines in vitro | NO | NO | [28] |
 RAF inhibitor 7a | - | 7a exhibited activities on A375P and WM3629 (IC50 0.62 μM and 4.49 μM) | NO | NO | [29] |
 RAF inhibitor 10d | 38.6 nM - CRAF 9.45 μM - BRAF WT | 10d exhibited activities on A375P and WM3629 (IC50 15.93 μM and 0.65 μM) | NO | NO | [30] |
RAF RBD-RAS binding inhibitors | |||||
 Kobe0065 | 46 ± 13 μM - KRAS G12V | Kobe0065 exhibits inhibitory activity toward HRas-CRAF binding | Kobe0065 (80-160 mg/kg, orally) inhibits activity on SW480 CDX harboring the KRAS G12V mutation | NO | [31] |
 Kobe2602 | 149 μM - KRAS G12V | Kobe2602 exhibits inhibitory activity toward HRas-CRAF binding | Kobe2602 (80 mg/kg, orally) exhibits antitumor activity on SW480 CDX harboring the KRAS G12V mutation | NO | [31] |
 MCP110 | - | MCP110 (20 μM) significantly inhibits Ras-mediated stimulation of CRAF activity in fibrosarcoma HT1080 cells | NO | NO | [32] |
CRAF Scaffold/chaperone protein inhibitors | |||||
 KBU2046 | HSP90 | KBU2046 (10 μmol/L) inhibits intracellular activation of CRAF, thereby achieving selective inhibition of cell motility. | KBU2046 (150 mg/kg, orally) with ZA (100 μg/kg, IP) targeting strategy | NO | [33] |
 Novobiocin | HSP90 | Novobiocin (0.8 mM) displayed a reduced cellular CRAF activity but not BRAF V600E | NO | Alterations of DNA Repair genes in solid neoplasm, NCT05687110, Phase 1, Recruiting | [34] |
 17-DMAG | 62 ± 29 nM - HSP90 | 17-DMAG (1 μM) reduces the kinase activity of CRAF and BRAF V600E | 17-DMAG (10 or 20 mg/kg, IP) in Prostatic cancer | NCT00803556, Phase 1, Completed; NCT00089362, Phase 1, Completed; NCT00248521, Phase 1 Active, not recruiting | [34] |
 Locostatin | RKIP | Locostatin (200 µM) binds RKIP protein and disrupts the interaction between RKIP, CRAF, and GRK2 | NO | NO | [35] |
Pan-RAF dimers selective inhibitors | |||||
 RAF709 | 0.5 nM - CRAF 0.4 nM - BRAF | RAF709 stabilizes BRAF-CRAF dimers (EC50 0.8 μM), inhibition proliferation of Calu-6 cells (EC50 0.95 μM) | RAF709 (30-200 mg/kg, orally) results in Calu-6 tumor regression | NO | [36] |
 TAK-632 | 1.4 nM - CRAF, 2.4 nM - BRAF V600E 8.3 nM- BRAF WT | TAK-632 shows antiproliferative effects both in A375 (GI50 of 40-190 nM) and SK-MEL-2 (GI50 of 190-250 nM) cells | TAK-632 (60-120 mg/kg, orally) exhibits an antitumor effect without toxicity in SK-MEL-2 melanoma | NO | [37] |
 LXH254 | 0.072 nM - CRAF 0.21 nM - BRAF 6.4 nM - ARAF | LXH254 (0-10 µM) inhibits both monomeric and dimeric RAF and promotes RAF dimer formation; More sensitivity to ARAF depletion cells | LXH254 (100 mg/kg, orally) decreased tumor-harboring BRAF mutations with or without activated NRAS or KRAS | NCT04294160, Phase 1, BRAF V600 Colorectal Cancer, Active, not recruiting; NCT02607813, Phase 1, NSCLC/Ovarian Cancer/Melanoma/Solid Tumors, Terminated; NCT02974725, Phase 1, NSCLC, Active, not recruiting | |
 LY3009120 | 15 nM - CRAF 5.8 nM - BRAF V600E 9.1 nM- BRAF WT | LY3009120 exhibits anti-proliferative effects on cell lines harboring BRAFV600E, KRASG13 and KRASG12 mutations | LY3009120 (20 mg/kg, orally) inhibits BRAF and KRAS mutant CRC CDX; (15 or 30 mg/kg, orally) in the H2405 model | NCT02014116, Phase 1, Advanced or Metastatic Cancer, Terminated | |
 Belvarafenib | 5 nM - CRAF 56 nM - BRAF WT 7 nM - BRAF V600E | Belvarafenib effect in BRAF- and NRAS-mutant tumors, but acquired ARAF mutations drive resistance | Belvarafenib reduced tumor burden in mice with A375SM melanoma. | NCT04835805, Phase 1, NRAS mutant Advanced Melanoma; NCT03118817, Phase 1, Solid Tumor; NCT04589845, Phase II, Solid Tumors; NCT02405065, Phase 1, Neoplasms | [42] |
CRAF Mult-kinase inhibitors | |||||
 Sorafenib | 6 nM - CRAF 20 nM - BRAF 15 nM - VEGFR3 20 nM - PDGFRβ 57 nM - FLT3 58 nM - c-Kit | Sorafenib (0.01 to 3 μM) blocks MAPK pathway with MEK 1/2 and ERK 1/2 phosphorylation (IC50, 40 and 100 nM, respectively) | Sorafenib (30-60 mg/kg, orally) produces broad spectrum antitumor activity in colon, breast, and non-small-cell lung cancer xenograft models | NCT04387695, Phase 3, Unresectable Hepatocellular Carcinoma|Portal Vein Thrombosis; NCT03456401, Phase 2, Renal Cancer; NCT01715441, Phase 2, Metastatic Colorectal Cancer With KRAS Mutation | |
 RAF265 | RAF/VEGFR2 | RAF265 inhibit cell viability of HT29 and MDAMB231 cells (IC50 values of 5 to 10 μM) | RAF265 (30 mg/kg qd, single use) and combination with RAD001 (both 12 mg/kg qd) in HCT116 xenografts | NCT00304525, Phase 1/2, Metastatic Melanoma;NCT01352273, Phase 1, Advanced Solid Tumors | |
 Avutometinib | 8.2 nM - BRAF V600E 56 nM - CRAF 160 nM - MEK 190 nM - BRAF | Avutometinib inhibits activation of ERK2 by MEK1 (IC50 of 160 nM) and activation of MEK1 by CRAF (IC50 of 56 nM) | Single or in combination with PD0325901 in HCT116 (KRAS-mutant) models, the ED50 for Avutometinib and PD0325901 are 0.056 and 0.80 mg/kg, respectively | NCT05669482 (Phase 1/2), KRAS Activating Mutation, Metastatic Cancer, Pancreas Cancer, Neoplasms Pancreatic Malignant Neoplasm of Pancreas | [46] |
 Regorafenib | 2.5 nM - CRAF 13/4.2/46 nM - VEGFR1/2/3 22 nM - PDGFRβ 7 nM - Kit 1.5 nM - RET | Regorafenib (0-10 μM) exhibits anti-proliferation activity in GIST 882, Thyroid TT, MDA-MB-231, HepG2, A375, and SW620 cells; as well as in Hep3B ( IC50 of 5 μM) | Regorafenib (10 mg/kg, Orally) inhibits rat GS9L glioblastoma model; (0-100 mg/kg, Orally) exhibits antitumorigenic and antiangiogenic effects in the Colo-205, MDA-MB-231, and 786-O model | NCT03465722, Phase 3, GIST; NCT01774344, Phase 3, Carcinoma, Hepatocellular; NCT02788279, Phase 3, Colorectal Cancer; NCT01271712, Phase 3, Gastrointestinal Stromal Tumors; NCT01103323, Phase 3, Metastatic Colorectal Cancer | |
 Erianin | CRAF/MEK | Erianin exhibits anti-proliferation effect in A375 (12.0 ± 0.9 nM), SK-MEL-28 (50.6 ± 1.7 nM), SK-MEL-2 (59.7 ± 7.2 nM) and HCT116 (20.6 ± 2.2 nM) | Erianin (50 mg/kg, Orally) inhibits A375, SK-MEL-28, SK-MEL-2 and HCT116 xenografts and melanoma/CRC patient derived tumor xenografts | NO | [50] |