CNP200137的发现：一种具有卓越临床前特性并对获得性耐药具有抗白血病活性的第二代menin-MLL抑制剂

Targeting the menin-mixed lineage leukemia (MLL, also known as KMT2A) interaction is a promising therapeutic strategy for treating acutemyeloid leukemia (AML). Oncogenic KMT2A-fusion proteins in AML recruit menin to chromatin, a critical step for activating leukemogenic target genes such as MEIS1 and HOX clustergenes. This menin-dependent leukemogenic program is also shared by mutant nucleophosmin 1 (NPM1) AML. Menin inhibitors recently demonstrated substantial clinical activity inKMT2A-rearranged (KMT2A-r) and NPM1-mutant (NPM1m) leukemias, as well as broader subsets exhibiting menin-dependent biology-collectively representing approximately 40% of AML cases. However, the emergence of on-target MEN1 resistance mutations has limited the long-term therapeutic benefits of first-generation agents such as revumenib and ziftomenib. Herein, we report the discovery of CNP200137, a next-generation menin-MLL inhibitor designed to effectively overcome MEN1-mediated resistance.Menin-MLL disruption was measured using and HTRF assay. Anti-proliferative activity was evaluated in KMT2A-r AML cells (MV4-11, MOLM-13), NPM1m AML cells (OCI-AML3), MLL wild type cells (MOLT-4), CRISPR-engineered MEN1-resistant mutants (M327I, T349M, G331R, E368K), and revumenib-resistant MOLM-13 cells. Cellular target engagement was assessed using qPCR, immunoblotting, and Astral-based global proteomics, which sensitively quantified robust menin protein degradation. Following pharmacokinetic (PK) characterization in mice, in vivo anti-tumor efficacy in mice bearing parental MV4-11 and resistant MV4-11(M327I) mutant was evaluated. The novel menin-MLL inhibitor CNP200137 effectively disrupted the menin-MLL interaction with nanomolar potency, exhibiting an IC₅₀ value of 2 nM against wild-type MEN1 and maintaining strong activity against the clinically relevant resistance mutants M327I with an IC₅₀ value of 13 nM and T349M with an IC₅₀ value of 11 nM. CNP200137 selectively exhibited strong anti-proliferative activity against AML cells harboring KMT2A rearrangements, with IC₅₀ values of 19 nM and 35 nM in MV4-11 and MOLM-13 cells, respectively, and an IC₅₀ value of 31 nM in NPM1-mutant OCI-AML3 cells. Notably, CNP200137 displayed superior anti-leukemic activity against both engineered MV4-11 cells with clinically relevant acquired resistance and revumenib-resistant MOLM-13 cells, compared to approved or clinical-stage menin inhibitors. Notably, CNP200137 displayed superior anti-leukemic activity against engineered MV4-11 cells with clinically relevant acquired resistance, compared to approved or clinical-stage menin inhibitors. Proteomic analysis revealed that these inhibitors induced stronger and more consistent degradation of menin proteins in MV4-11, MOLM-13 and OCI-AML3 cells, with the downregulation of MEIS1 genes. In mice, the compounds exhibited an improved PK profile and strongly suppressed tumor growth in both parental and resistant MV4-11 xenografts. This novel, potent, and orally available second-generation menin-MLL inhibitor, CNP200137, represents a promising therapeutic strategy for overcoming acquired resistance in KMT2A-rearranged and NPM1-mutant acute leukemias.