ALK (anaplastic lymphoma kinase) is a tyrosine kinase that is constitutively activated in certain cancers, following gene alterations such as chromosomal translocation, amplification or point mutation. Lorlatinib is a dual inhibitor of ALK and ROS1 with Ki values of 2nM, 7nM and 70nM for ROS1, ALK and ALK-L1196M (measured by kinase screening assays). Cellular study showed that Lorlatinib possessed a more potent kinase inhibitory effect on EML4-ALK mutation, compared with Crizotinib, with IC50 values on p-ALK of 1.3nM, 0.2nM, 1.6nM, 15nM, 4.2nM, 21nM, 9nM, 77nM and 38nM for different mutations ALK, F1174L, C1156Y, G1269A, S1206Y, L1196M, L1152R, G1202R and 1151Tins in 3T3-EML4-ALK engineered cell lines[1]. As a dual inhibitor, Lorlatinib also showed potency to block crizotinib-resistant ROS1 mutations. In vitro study showed that Lorlatinib exhibited subnanomolar IC50 values of ranging in 0.19–0.53nM against pROS1-Tyr2274 in a panel of NIH 3T3 and BaF3 cells engineered to express selected oncogenic ROS1 fusion variants including CD74-ROS1, SLC34A2-ROS1, and FIGROS1. Also, it inhibited the crizotinib-refractory ROS1-G2032R mutation and the ROS1-G2026M gatekeeper mutation. In vivo studies showed that Lorlatinib markedly inhibit the tumor growth in models expressing FIG-ROS1 (1, 3mg/kg), CD74-ROS1 (0.2-6mg/kg) and the CD74-ROS1-G2032R (3-60mg/kg) mutation[2]. In addition, Lorlatinib is designed for CNS availability for a single 10mg/kg oral dose of this compound achieved its distributes into the CNS in rat[1]. Lorlatinib at dose of 10mg/kg (but administered by s.c. osmotic pumps) demonstrated the antitumor activity in a genetically engineered mouse model of FIG-ROS1 glioblastoma[2].
作用机制
Lorlatinib is an ATP-competitive small-molecule inhibitor.[2]
Lorlatinib/劳拉替尼 细胞实验
Cell Line
Concentration
Treated Time
Description
References
H3122
1μM
1 hour
Identify GUK1 as a metabolic target of ALK signaling
Cell. 2025 Mar 6;188(5):1248-1264. e23
CLB-GAR
30 nM
6 hours
To investigate the effect of ALK inhibition on CHK1 phosphorylation, results showed lorlatinib treatment decreased CHK1 S280 phosphorylation.
Proc Natl Acad Sci U S A. 2024 Jan 2;121(1):e2315242121.
CLB-GE
30 nM
6 hours
To investigate the effect of ALK inhibition on CHK1 phosphorylation, results showed lorlatinib treatment decreased CHK1 S280 phosphorylation.
Proc Natl Acad Sci U S A. 2024 Jan 2;121(1):e2315242121.
MGH919–5
1μM
1 hour
Identify GUK1 as a metabolic target of ALK signaling
Cell. 2025 Mar 6;188(5):1248-1264. e23
MGH045–1
1μM
1 hour
Identify GUK1 as a metabolic target of ALK signaling
Through ENU mutagenesis screening, lorlatinib-resistant ALK compound mutations were identified, including compound mutations of I1171N and G1202R.
EBioMedicine. 2019 Mar;41:105-119.
Ba/F3 cells
10, 100, 300, 600, 1000 nM
3 h
The inhibitory effect of lorlatinib on the ALK-G1202R + G1269A double mutation was evaluated, and the results showed that this mutation was resistant to lorlatinib.
EBioMedicine. 2019 Mar;41:105-119.
NCI-H3122
4 µM
2–4 months
To study the evolution of resistance to different ALK inhibitors in NCI-H3122 cells, it was found that Lorlatinib could select for cells with strong resistance to high concentrations of ALK inhibitors.
Nat Commun. 2020 May 14;11(1):2393.
JFCR-028-3-LR1000#1
1 or 3 µM
1 week
To establish Lorlatinib-resistant cell models and study their cross-resistance to Lorlatinib and other ALK-TKIs.
NPJ Precis Oncol. 2022 Mar 17;6(1):16.
JFCR-028-3 cells
100 nM
9 days
To evaluate the effect of Lorlatinib on JFCR-028-3 cells, it was found that MIG6 depletion increased resistance to ALK-TKIs.
JCI Insight. 2023 Dec 22;8(24):e173688.
TNBC cells (acquired PARP inhibitor resistance)
0.5 µM
24 h
Evaluate the effect of CDK9-IN-2 on the interaction between ALK and CDK9 in TNBC cells (acquired PARP inhibitor resistance), showing that ALK inhibitor reduced the binding signal of p-ALK and CDK9.
Nat Cancer. 2022 Oct;3(10):1211-1227
OVCA433 ovarian cancer cells
0.5 µM
24 h
Evaluate the effect of CDK9-IN-2 on the interaction between ALK and CDK9 in OVCA433 cells, showing that ALK inhibitor reduced the binding signal of p-ALK and CDK9.
Nat Cancer. 2022 Oct;3(10):1211-1227
SKOV3 ovarian cancer cells
0.5 µM
24 h
Evaluate the effect of CDK9-IN-2 on the interaction between ALK and CDK9 in SKOV3 cells, showing that ALK inhibitor reduced the binding signal of p-ALK and CDK9.
Nat Cancer. 2022 Oct;3(10):1211-1227
Lorlatinib/劳拉替尼 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
BALB-c nu/nu mice
JFCR-041-2 cell xenograft model
Oral
10 mg/kg
Once daily for 5-6 days/week
Investigate lorlatinib resistance mechanisms
EBioMedicine. 2019 Mar;41:105-119.
Mouse
EML4-ALK L1196M
Intraperitoneal injection
0.5, 2, 10 mg/kg
Daily for 2 weeks
Lorlatinib suppressed ALK-driven tumor growth in a dose-dependent manner, but LUSC showed significant resistance to lorlatinib treatment
J Exp Med. 2024 Mar 4;221(3):e20232028
BALB/c nu/nu mice
Mouse model of EML4-ALK-G1202R mutation
Oral
10, 30, 50 mg/kg
Once daily for 5-6 days/week
In the mouse model of EML4-ALK-G1202R mutation, lorlatinib initially significantly shrank the tumor, but the tumor eventually regrew, and the ALK-G1202R + G1269A double mutation was identified.
EBioMedicine. 2019 Mar;41:105-119.
Mouse
EML4-ALK L1196M model
Intraperitoneal injection
0.5, 2, 10 mg/kg
Daily for 2 weeks
To investigate the inhibitory effect of Lorlatinib on tumors in the EML4-ALK L1196M mouse model, it was found that Lorlatinib dose-dependently suppressed ALK-driven tumor growth, with LUAD showing significant decrease in tumor sizes at 0.5 and 2 mg/kg Lorlatinib treatments, while LUSC showed no significant change.
J Exp Med. 2024 Mar 4;221(3):e20232028
NOD/SCID gamma mice
Subcutaneous xenograft model
Oral
3 mg/kg
Once daily, 4 days on, 3 days off
Evaluate the inhibitory effect of lorlatinib alone or in combination with selumetinib on ROS1 fusion-positive tumors.
Clin Cancer Res. 2020 Jun 15;26(12):2932-2945
Mice
NSG mice
Oral
6 mg/kg
Daily administration for 3 weeks
To evaluate the therapeutic efficacy of Lorlatinib in ALK-positive NSCLC in vivo, it was found that Lorlatinib significantly inhibited tumor growth.
Nat Commun. 2020 May 14;11(1):2393.
BALB/c nude mice
JFCR-028-3 cell xenograft model
Oral
10 mg/kg
5 days per week for 4 weeks
To evaluate the antitumor effect of Lorlatinib in vivo on JFCR-028-3 cell xenograft model, it was found that MIG6 depletion conferred resistance to ALK-TKIs, and combination therapy could overcome the resistance.
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