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Chemical Structure| 1070773-09-9 Chemical Structure| 1070773-09-9

Structure of RK-33
CAS No.: 1070773-09-9

Chemical Structure| 1070773-09-9

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RK-33 is a first-in-class small molecule inhibitor of DDX3 (a RNA helicase) and causes G1 cell cycle arrest, induces apoptosis, and promotes radiation sensitization in DDX3-overexpressing cells.

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Product Citations

Product Citations

Dian, Meijuan ; Yun, Liang ; Meng, Qingyu ; Lin, Songwen ; Ji, Ming ; Zhou, Ying , et al.

Abstract: Approximately 30% of human cancers carry various mutations, including KRAS, NRAS, and HRAS. Among these mutations, KRAS is the most prevalent isoform detected in lung cancer. While several small molecular inhibitors targeting specifically KRASG12C have been developed and tested clinically, alternative approaches are still necessary due to expected drug resistance. In this study, we present evidence that the loss of DDX3X significantly delays tumor progression in various KRAS-driven lung cancer models. Inhibition of DDX3X disrupts cysteine and glutathione metabolism, thereby inducing in lung cancer cells. This effect is primarily mediated by the downregulation of Cystathionine-β-synthase (CBS), the rate-limiting enzyme in cysteine generation. Mechanistically, DDX3X directly binds to the transcription factor JUND, which mediates the transcriptional regulation of METTL16, a key N6-methyladenosine methyltransferase, and subsequently regulates m6A modification and translation of CBS transcripts. This cascade induces hypermethylation and high expression of CBS, consequently triggering cysteine production and maintaining antioxidative homeostasis, which is essential for the survival of KRAS-driven lung cancer cells. Finally, we demonstrate that a newly developed DDX3X degrader J10 efficiently delays lung cancer progression with multiple advantages compared to DDX3X small molecular inhibitor and limited side effects. These findings unveil the potential of DDX3X as a valuable target for adjuvant therapies in managing KRAS-driven lung cancer.

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Product Details of RK-33

CAS No. :1070773-09-9
Formula : C23H20N6O3
M.W : 428.44
SMILES Code : O=C1N(CC2=CC=C(OC)C=C2)C3=NC=NC4=C(N=CN4CC5=CC=C(OC)C=C5)C3=N1
MDL No. :MFCD30489740
InChI Key :COUMZXFUZDBRCZ-UHFFFAOYSA-N
Pubchem ID :46184988

Safety of RK-33

GHS Pictogram:
Signal Word:Warning
Hazard Statements:H302-H315-H319-H335
Precautionary Statements:P261-P305+P351+P338

Related Pathways of RK-33

DNA

Isoform Comparison

Biological Activity

Target
  • helicase

In Vitro:

Cell Line
Concentration Treated Time Description References
U2OS human osteosarcoma cells 6 µM 1 hour To evaluate the effect of RK-33 on SG assembly, the results showed that RK-33 significantly inhibited the assembly of G3BP1-positive SGs. Biochem Pharmacol. 2020 Dec;182:114280.
RD cells 1 µM 1 hour To study the effect of RK-33 on HCoV-OC43 infection, results showed that RK-33 significantly reduced viral titers. Front Microbiol. 2022 Aug 25;13:959577.
Vero cells 0.01, 0.04, 0.16, 0.63, 1.25, 2.5, 10, 20, 50 µM 22 hours To test the inhibitory effect of RK-33 on various viruses, results showed that RK-33 effectively inhibited the replication of DENV-2, ZIKV, WNV, RSV, and hPIV-3. Cells. 2020 Jan 9;9(1):170.
MDA-MB-435 cells 4.5 µM 24 hours To evaluate changes in the protein landscape after RK-33 treatment, changes in mitochondrial translation, cell division, and cell cycle-related proteins were observed. Transl Oncol. 2018 Jun;11(3):755-763.
MCF7 cells 1.5 µM, 3 µM, 4.5 µM 24 hours To evaluate the effect of RK-33 on the cell cycle using the FUCCI system, a delay in all cell cycle phases was observed. Transl Oncol. 2018 Jun;11(3):755-763.
Calu-3 cells 5 µM 24 hours To study the effect of RK-33 on SARS-CoV-2 infection, results showed that RK-33 significantly reduced viral load and downregulated most SARS-CoV-2 gene expressions. Front Microbiol. 2022 Aug 25;13:959577.
SW1990 SP cells 4.32 µM (IC50) 48 hours To evaluate the effect of RK-33 on cancer stem cell markers, results showed that RK-33 significantly downregulated the expression of CD44v6, NANOG, SOX9, and β-CATENIN. Gastroenterology. 2020 Nov;159(5):1898-1915.e6.
Capan1 SP cells 5.53 µM (IC50) 48 hours To evaluate the effect of RK-33 on cancer stem cell markers, results showed that RK-33 significantly downregulated the expression of CD44v6, NANOG, SOX9, and β-CATENIN. Gastroenterology. 2020 Nov;159(5):1898-1915.e6.
9–26 NP fibroblasts 9.23 µM (IC50) 48 hours To evaluate the effect of RK-33 on normal fibroblasts, results showed that RK-33 had lower toxicity to normal cells. Gastroenterology. 2020 Nov;159(5):1898-1915.e6.
Human lung fibroblasts 1 or 10 µM 48 hours RK-33 inhibited TGF-β1-induced upregulation of NEU3 JCI Insight. 2023 Apr 10;8(7):e167566.
A549 cells (high DDX3 expression) 1 µM and 2 µM 72 hours To evaluate the cytotoxicity of RK-33 on lung cancer cells, results showed that cells with high DDX3 expression were more sensitive to RK-33 EMBO Mol Med. 2015 May;7(5):648-69.
H3255 cells (low DDX3 expression) 1 µM and 2 µM 72 hours To evaluate the cytotoxicity of RK-33 on lung cancer cells, results showed that cells with high DDX3 expression were more sensitive to RK-33 EMBO Mol Med. 2015 May;7(5):648-69.
MCF10A 7.4 µM (IC50) 72 hours To evaluate the cytotoxicity of RK-33 on normal breast cells MCF10A, results showed that MCF10A was less sensitive to RK-33 Oncogene. 2018 Jan 4;37(1):63-74.
MCF7 2.8–4.5 µM (IC50) 72 hours To evaluate the cytotoxicity of RK-33 on breast cancer cells MCF7, results showed that MCF7 was more sensitive to RK-33 Oncogene. 2018 Jan 4;37(1):63-74.
MDA-MB-231 2.8–4.5 µM (IC50) 72 hours To evaluate the cytotoxicity of RK-33 on breast cancer cells MDA-MB-231, results showed that MDA-MB-231 was more sensitive to RK-33 Oncogene. 2018 Jan 4;37(1):63-74.
MDA-MB-435 2.8–4.5 µM (IC50) 72 hours To evaluate the cytotoxicity of RK-33 on breast cancer cells MDA-MB-435, results showed that MDA-MB-435 was more sensitive to RK-33 Oncogene. 2018 Jan 4;37(1):63-74.
Rat primary cortical neurons 1 µM 72 hours To validate the neuroprotective effects of RK-33 against the combined neurotoxicity of HIV Tat and cocaine, results showed RK-33 significantly reduced neuronal death J Neuroimmune Pharmacol. 2020 Jun;15(2):209-223.
Marc-145 cells 8.495 µM 72 hours RK-33 inhibits DDX3X expression and reduces PRRSV replication. Vet Res. 2024 Aug 18;55(1):103.
DAOY cells 2.5 µM 72 hours RK-33 inhibited the growth and promoted cell death in DAOY cells with an IC50 value of 2.5 μM. Transl Oncol. 2019 Jan;12(1):96-105.
UW228 cells 3.5 µM 72 hours RK-33 inhibited the growth and promoted cell death in UW228 cells with an IC50 value of 3.5 μM. Transl Oncol. 2019 Jan;12(1):96-105.
A549, H1299, H23, and H460 cells 4.4–8.4 µM (IC50) To evaluate the cytotoxicity of RK-33 on lung cancer cells, results showed that cells with high DDX3 expression were more sensitive to RK-33 EMBO Mol Med. 2015 May;7(5):648-69.
H3255 cells > 25 µM (IC50) To evaluate the cytotoxicity of RK-33 on lung cancer cells, results showed that cells with high DDX3 expression were more sensitive to RK-33 EMBO Mol Med. 2015 May;7(5):648-69.
DU145 3 µM (IC50) RK-33 inhibited proliferation and induced G1 phase cell-cycle arrest in DU145 cells. Cancer Res. 2016 Nov 1;76(21):6340-6350.
LNCaP 6 µM (IC50) RK-33 inhibited proliferation and induced G1 phase cell-cycle arrest in LNCaP cells. Cancer Res. 2016 Nov 1;76(21):6340-6350.
22Rv1 3–6 µM (IC50) RK-33 inhibited proliferation and induced G1 phase cell-cycle arrest in 22Rv1 cells. Cancer Res. 2016 Nov 1;76(21):6340-6350.
PC3 12 µM (IC50) RK-33 had minimal effects on PC3 cells, with no significant changes in cell cycle. Cancer Res. 2016 Nov 1;76(21):6340-6350.

In Vivo:

Species
Animal Model
Administration Dosage Frequency Description References
Mice Twist1/KrasG12D autochthonous lung tumor model Intraperitoneal injection 20 mg/kg Twice a week for 7 weeks To evaluate the effect of RK-33 in combination with radiation, results showed that RK-33 significantly enhanced radiation-induced tumor regression EMBO Mol Med. 2015 May;7(5):648-69.
C57BL/6 mice Bleomycin-induced pulmonary fibrosis model Intraperitoneal injection 20 mg/kg Every 2 days for 11 days RK-33 potentiated survival, reduced lung inflammation and fibrosis, and decreased tissue levels of DDX3, TGF-β1, and NEU3 JCI Insight. 2023 Apr 10;8(7):e167566.
SCID mice DU145-Luc xenograft model Intraperitoneal injection 50 mg/kg Thrice weekly for two weeks The combination of RK-33 and radiation significantly reduced tumor growth and induced more apoptosis. Cancer Res. 2016 Nov 1;76(21):6340-6350.
Nude mice Medulloblastoma xenograft model Intraperitoneal injection 50 mg/kg Every alternate day for two weeks The combination of RK-33 and 5 Gy radiation caused tumor regression in a mouse xenograft model of medulloblastoma. Transl Oncol. 2019 Jan;12(1):96-105.

Protocol

Bio Calculators
Preparing Stock Solutions 1mg 5mg 10mg

1 mM

5 mM

10 mM

2.33mL

0.47mL

0.23mL

11.67mL

2.33mL

1.17mL

23.34mL

4.67mL

2.33mL

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