ML216 (CID-49852229) acts as a powerful, selective, and cell-penetrable blocker of BLM helicase's DNA unwinding function, exhibiting IC50 values of 2.98 μM for BLM full-length and 0.97 μM for BLM636-1298. Additionally, ML216 suppresses BLM's ssDNA-dependent ATPase activity with a Ki of 1.76 µM, displaying antitumor activity[1][2].
体内研究
While ML216 similarly inhibits the unwinding activity of the related BLM and WRN helicases in vitro, its biological effects in human cells predominantly rely on BLM, indicating a BLM-specific mechanism of action in vivo. An elucidating co-crystal structure of BLM bound to the inhibitor would be revealing. It's suggested that cellular conditions in vivo may prompt a WRN conformation resistant to ML216[2].
体外研究
ML216 treatment (12.5-50 μM; 24-72 hours; PSNG5 and PSNG13 cells) reduces PSNF5 cell proliferation in a dose-dependent fashion, yet does not affect PSNG13 cells[1].
ML216 treatment results in a significant rise in the frequency of sister chromatid exchanges (SCEs) in PSNF5 cells, without affecting PSNG13 cells[1].
ML216 enhances the susceptibility of PSNF5 cells to aphidicolin while having no sensitizing impact on the BLM-lacking isogenic PSNG13 cells[1].
ML216 suppresses both the full-length WRN (IC50 of 5 μM) and the truncated WRN500-946 (IC50 of 12.6 μM), with the former showing 2.5 times greater sensitivity to ML216's inhibitory effect. Compared to WRN, BLM exhibits slightly higher sensitivity to ML216 inhibition (1.7 times based on IC50 values). Although ML216 effectively inhibits WRN, it predominantly targets BLM in human cells. ML216 equally inhibits the proliferation of both WRN+ and WRN-deficient cells and likewise increases their sensitivity to aphidicolin[1].
ML216 细胞实验
Cell Line
Concentration
Treated Time
Description
References
XG2 cell line
4.9 µM
48 and 96 hours
ML216 induced cell cycle arrest and apoptosis
Front Immunol. 2022 Dec 9;13:983181
XG19 cell line
1.2 µM
48 and 96 hours
ML216 induced cell cycle arrest and apoptosis
Front Immunol. 2022 Dec 9;13:983181
S18 ES cells (BL6 × SPRET F1 hybrid)
25 μM
5, 10, or 21 days
induced recombination for genetic mapping, identified Hprt as the gene underlying 6-TG susceptibility
Proc Natl Acad Sci U S A. 2018 Apr 3;115(14):3680-3685
BL6 × CAST F1 hybrid ES cells
25 μM
induced mitotic recombination to generate homozygous genotypes
Proc Natl Acad Sci U S A. 2018 Apr 3;115(14):3680-3685
mouse embryonic stem (ES) cells
25 μM
1 or 5 days
inhibited BLM DNA helicase, induced random mitotic cross-overs, elevated homologous recombination rates
Proc Natl Acad Sci U S A. 2018 Apr 3;115(14):3680-3685
CH3+5 HCT116 cells
8 μM
72 hours
To evaluate the effect of ML216 on MMR-corrected HCT116 cells, results showed that ML216 had a weaker inhibitory effect on the growth of CH3+5 HCT116 cells.
Proc Natl Acad Sci U S A. 2022 Dec 20;119(51):e2211775119
HCT116 cells
8 μM
72 hours
To evaluate the inhibitory effect of ML216 on MSI CRC cells, results showed that ML216 significantly inhibited the growth of HCT116 cells and induced PUMA expression and apoptosis.
Proc Natl Acad Sci U S A. 2022 Dec 20;119(51):e2211775119
human myeloma cell lines (HMCLs)
0.78 µM to 100 µM
4 days
Evaluation of the inhibitory effect of ML216 on HMCLs proliferation, with a median IC50 of 2.78 µM
Front Immunol. 2022 Dec 9;13:983181
PC3 cells
5 μM
48 hours
To evaluate the inhibitory effects of ML216 combined with olaparib on the proliferation, clonogenicity, invasion, and migration of PC3 cells. Results showed that the combination treatment had stronger inhibitory effects compared to monotherapy.
J Transl Med. 2023 Jul 6;21(1):445
A549 cells
2 µM
3 or 5 days
To evaluate the proliferation inhibition and radiosensitization effects of ML216 on A549 cells. Results showed ML216 combined with olaparib significantly enhanced the radiosensitivity of A549 cells.
Cancer Biol Med. 2021 Dec 1;19(8):1150–71
H1299 cells
2-10 µM
3 or 5 days
To evaluate the proliferation inhibition and radiosensitization effects of ML216 on H1299 cells. Results showed ML216 significantly enhanced the radiosensitivity of H1299 cells.
Cancer Biol Med. 2021 Dec 1;19(8):1150–71
H460 cells
2-10 µM
3 or 5 days
To evaluate the proliferation inhibition and radiosensitization effects of ML216 on H460 cells. Results showed ML216 significantly enhanced the radiosensitivity of H460 cells.
Cancer Biol Med. 2021 Dec 1;19(8):1150–71
IMR-90 cells
50 μM
24 hours
To evaluate the effect of ML216 on DNA damage-induced senescence, results showed that ML216 significantly reduced the number of senescent cells.
Cells. 2022 Dec 29;12(1):145
293T cells
0, 20, 50, or 100 μM
12 hours
To evaluate the effect of ML216 on DBC1-BLM interaction, results showed that ML216 concentration-dependently increased the binding of BLM to DBC1.
Cells. 2022 Dec 29;12(1):145
WPMY-1 cells
10 μM
48 hours
Evaluate the inhibitory effect of ML216 and CDDP on WPMY-1 cell proliferation, showing no synergistic effect in combination therapy
Molecules. 2022 Dec 12;27(24):8790
LNCap cells
10 μM
48 hours
Evaluate the inhibitory effect of ML216 and CDDP on LNCap cell proliferation, showing that combination therapy has a stronger inhibitory ability
Molecules. 2022 Dec 12;27(24):8790
22RV1 cells
10 μM
48 hours
Evaluate the inhibitory effect of ML216 and CDDP on 22RV1 cell proliferation, showing that combination therapy has a stronger inhibitory ability
Molecules. 2022 Dec 12;27(24):8790
PC3 cells
10 μM
48 hours
Evaluate the inhibitory effect of ML216 and CDDP on PC3 cell proliferation, showing that combination therapy has a stronger inhibitory ability
Molecules. 2022 Dec 12;27(24):8790
Plasmodium falciparum Dd2
1 nM to 1,000 μM
48 hours
To evaluate the inhibitory effect of ML216 on parasite growth
mSphere. 2020 Nov 25;5(6):e00956-20
XG1 cell line
13.2 µM
48 and 96 hours
ML216 induced cell cycle arrest and apoptosis
Front Immunol. 2022 Dec 9;13:983181
ML216 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Nude mice
HCT116 xenograft tumors
Intraperitoneal injection
1.5 mg/kg
Daily for the first 7 days and then every other day for 13 days
To evaluate the therapeutic effect of ML216 on MSI CRC xenograft tumors, results showed that ML216 significantly suppressed the growth of HCT116 tumors and increased TUNEL and active caspase 3 staining.
Proc Natl Acad Sci U S A. 2022 Dec 20;119(51):e2211775119
BALB/c nude mice
PC3 xenograft model
Intraperitoneal injection
15 mg/kg/d
Three times a week for 3 weeks
To evaluate the inhibitory effects of ML216 combined with olaparib on the growth of PC3 xenograft tumors. Results showed that the combination treatment significantly reduced tumor volume and weight, with no observed toxicity in normal tissues.
J Transl Med. 2023 Jul 6;21(1):445
BABL/c Nu/Nu male nude mice
A549 xenograft model
Subcutaneous injection
25 mg/kg/d
3 continuous days
To evaluate the radiosensitization effects of ML216 combined with olaparib on A549 xenograft model. Results showed the combination significantly inhibited tumor growth and enhanced radiosensitivity.
Cancer Biol Med. 2021 Dec 1;19(8):1150–71
C57BL/6J mice
Naturally aged mouse model and bleomycin-induced idiopathic pulmonary fibrosis (IPF) mouse model
Intraperitoneal injection
1 mg/kg (naturally aged model) and 0.5 mg/kg (IPF model)
Twice a week for 5 weeks (naturally aged model) or 3 weeks (IPF model)
To evaluate the effect of ML216 on senescence and fibrosis in vivo, results showed that ML216 significantly reduced the number of senescent cells, improved pulmonary function, and decreased the expression of fibrosis markers.
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