Autophagy, the sequestration of organelles and proteins in autophagic vesicles (AVs) and degradation of this cargo through lysosomal fusion, allows tumor cells to survive metabolic and therapeutic stresses[2]. Lys05, a dimeric chloroquine, is a new lysosomal autophagy inhibitor which more potently accumulates in the lysosome and blocks autophagy compared with hydroxychloroquine (HCQ)[3]. C8161 xenografts matched for tumor size were treated with Lys05 (76 mg/kg; i.p.) for 48 h . Cells with intact nuclear and cytoplasmic membranes contained large AVs in Lys05-treated tumors. Meanwhile, significantly higher LC3II/LC3I levels were observed in Lys05-treated tumors compared with control- or HCQ-treated tumors, providing further evidence of in vivo autophagy inhibition. Mice bearing 1205Lu xenografts matched for tumor volume were treated with Lys05 (76 mg/kg; i.p.) and dosed for 3 d of daily treatment with 2 d off treatment. Lys05 treatment resulted in a 53% reduction in the average daily tumor growth rate compared with vehicle-treated controls. A significant six-fold accumulation of AV was observed at the end of 14 d of treatment in Lys05-treated tumors, compared with control group. Extensive tumor necrosis was observed in Lys05-treated tumors[2].
Lys05 细胞实验
Cell Line
Concentration
Treated Time
Description
References
MCF-7 cells
12.3 ± 0.7 µM
24 hours
Evaluate antiproliferative activity, results showed BAQ12 and BAQ13 had ~3-fold higher potency than Lys05
Nat Commun. 2020 Sep 15;11(1):4615
HEL cells
5 µM
Lys05 reduced HEL cell number independently of ruxolitinib inhibition.
Blood Cancer J. 2023 Jul 10;13(1):106
H460 cells
11.1 ± 0.6 µM
24 hours
Evaluate antiproliferative activity, results showed BAQ12 and BAQ13 had ~3-fold higher potency than Lys05
Nat Commun. 2020 Sep 15;11(1):4615
HT29 cells
10.6 ± 3.4 µM
24 hours
Evaluate antiproliferative activity, results showed BAQ12 and BAQ13 had ~3-fold higher potency than Lys05
Nat Commun. 2020 Sep 15;11(1):4615
BXPC-3 cells
7.8 ± 0.9 µM
24 hours
Evaluate antiproliferative activity, results showed BAQ12 and BAQ13 had ~3-fold higher potency than Lys05
Nat Commun. 2020 Sep 15;11(1):4615
PANC-1 cells
13.3 ± 2.0 µM
24 hours
Evaluate antiproliferative activity, results showed BAQ12 and BAQ13 had ~3-fold higher potency than Lys05
Nat Commun. 2020 Sep 15;11(1):4615
MIA PaCa-2 cells
16.7± 4.7 µM
24 hours
Evaluate antiproliferative activity, results showed BAQ12 and BAQ13 had ~3-fold higher potency than Lys05
Nat Commun. 2020 Sep 15;11(1):4615
A375P cells
0 – 1000 nM
2 weeks
To evaluate the effect of Lys05 on long-term clonogenic growth, results showed that Lys05 significantly suppressed long-term clonogenic growth of melanoma cells.
Cancer Discov. 2019 Feb;9(2):220-229
A375P cells
3 μM
6 hours
To evaluate the effect of Lys05 on lysosomal membrane permeabilization, results showed that Lys05 treatment increased lysosomal membrane permeabilization.
Cancer Discov. 2019 Feb;9(2):220-229
MDA-MB-231 cells
20 μM
72 hours
Evaluate Lys05's effect on pro-inflammatory cytokine expression; significantly increased Cxcl9, Cxcl10, Ccl5, Ifna, and Ifnb expression
Autophagy. 2024 Mar;20(3):525-540
Yumm 2.1 CTNNB1
1, 3, 5, 10μM
16-24 hours
To assess the sensitivity of Lys05 in β-catenin-stabilized melanoma cells
Cancer Res. 2017 Nov 1;77(21):5873-5885
Yumm 1.7 overexpressing WNT5A
1, 3, 5, 10μM
16-24 hours
To assess the sensitivity of Lys05 in Wnt5A-high and β-catenin-low melanoma cells
Cancer Res. 2017 Nov 1;77(21):5873-5885
Yumm 1.7
1, 3, 5, 10μM
16-24 hours
To assess the sensitivity of Lys05 in Wnt5A-low and β-catenin-high melanoma cells
Cancer Res. 2017 Nov 1;77(21):5873-5885
WM164
1, 3, 5, 10μM
16-24 hours
To assess the sensitivity of Lys05 in Wnt5A-low and β-catenin-high melanoma cells
Cancer Res. 2017 Nov 1;77(21):5873-5885
FS13
1, 3, 5, 10μM
16-24 hours
To assess the sensitivity of Lys05 in Wnt5A-high and low β-catenin melanoma cells
Cancer Res. 2017 Nov 1;77(21):5873-5885
HL-60 cells
5 µM
Lys05 modified neither the response of JAK2 WT HL-60 cells to ruxolitinib.
Blood Cancer J. 2023 Jul 10;13(1):106
SET-2 cells
5 µM
Lys05 strongly enhanced the cytotoxic effects of ruxolitinib in JAK2V617F cell lines.
Blood Cancer J. 2023 Jul 10;13(1):106
K562 cells expressing mRFP-GFP-LC3
10 µM
4 hours
To evaluate the inhibitory effect of Lys05 on autophagy flux, results showed that Lys05 treatment led to a significant accumulation of yellow fluorescence, indicating a complete block in autophagy flow.
Leukemia. 2019 Apr;33(4):981-994
K562 cells
1 µM and 10 µM
4 hours
To evaluate the inhibitory effect of Lys05 on autophagy, results showed that Lys05 at 1 µM and 10 µM concentrations significantly increased the level of membrane-bound LC3B-II, indicating increased accumulation of autophagosomes.
Leukemia. 2019 Apr;33(4):981-994
CD34+ CML cells
5 µM
To evaluate the inhibitory effect of Lys05 on autophagy, results showed that Lys05 treatment led to accumulation of LC3 puncta and increased levels of autophagy substrates SQSTM1/p62, NBR1, and NCOA4.
Leukemia. 2019 Apr;33(4):981-994
Lys05 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
NSG mice
HT29 colorectal cancer xenograft model
Intraperitoneal injection
10 mg/kg
Two daily doses
To evaluate the effect of Lys05 on tumor growth, results showed that two doses of Lys05 had only a transient effect on tumor growth.
Cancer Discov. 2019 Feb;9(2):220-229
Mice
Melanoma xenograft models and colon cancer xenograft model
Intraperitoneal injection
10 mg/kg to 80 mg/kg
Daily dosing or intermittent high-dose administration
Lys05 significantly blocks autophagy in vivo and exhibits single-agent antitumor activity. Compared to HCQ, Lys05 more effectively accumulates in and deacidifies lysosomes, leading to sustained autophagy inhibition and tumor growth suppression. High-dose Lys05 causes Paneth cell dysfunction and intestinal toxicity in mice.
Autophagy. 2012 Sep;8(9):1383-4
FVB mice
Transplanted PyMT-driven mammary tumor model
Intraperitoneal injection
50 mg/kg
Once daily for one week
Assess Lys05's impact on CD8+ T cell infiltration; significantly increased CD8+ T cell infiltration in tumors
To evaluate the inhibitory effect of Lys05 on melanoma tumor growth with different Wnt status
Cancer Res. 2017 Nov 1;77(21):5873-5885
NSG mice
HEL cell xenograft model
Intraperitoneal injection
32 mg/kg
Daily for 14 days
Lys05 improved the response to ruxolitinib and significantly prolonged the mice's overall survival.
Blood Cancer J. 2023 Jul 10;13(1):106
Nude mice
C8161 xenograft
Intraperitoneal injection
76 mg/kg
Once daily for 48 hours
Evaluate the in vivo autophagy inhibition and antitumor efficacy of Lys05. Results showed Lys05 significantly increased AV number and inhibited tumor growth.
Proc Natl Acad Sci U S A. 2012 May 22;109(21):8253-8
Wistar rats
HCC model
Intra-arterial administration
40 mg/kg
Single dose
Combination therapy coupling autophagy inhibition and TAE in a rat model of HCC resulted in a 21% increase in tumor necrosis compared with TAE alone
Radiology. 2017 Jun;283(3):702-710
Mice
Scl-tTa-BCR-ABL/GFP-LC3 mouse model
20 mg/kg/day
2 days or 7 days
To evaluate the inhibitory effect of Lys05 on autophagy, results showed that Lys05 treatment led to increased GFP-LC3 levels and accumulation of SQSTM1/p62, indicating autophagy inhibition. Additionally, Lys05 treatment reduced the number of LT-HSCs and promoted expansion of MPP and LSK cells.
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