Journal: Cell Death & Disease

Article Title: Simultaneous targeting of KRAS and CDK4 synergistically induces durable growth arrest in pancreatic cancer cells

doi: 10.1038/s41419-025-08362-w

Figure Lengend Snippet: A Heat map depicting DE genes according to the z-score after performing DeSeq2 analysis of four different samples (DMSO, 5 µM Palbociclib, 0.5 µM MRTX1133, or combination treatment, for 24 h (Day2), n = 3) in AsPC-1 cells. Only genes with |log2fold| ≥ 0.6, adjusted p-value (padj.) <0.05, and baseMean ≥15 were included in the analysis. Supplementary Table contains DE genes and normalized read counts. B Downregulated genes in Palbociclib + MRTX1133 (24 h treatment) vs. DMSO were correlated with the Molecular Signature Database (MSigDB) Hallmark 2020 and ChEA 2022 ( C ) datasets, using the Enrichr platform to identify potentially impaired pathways. Top 10 (ChEA 2022: human only), p-value ranked (−log10). D Gene set enrichment analysis (GSEA) of combination treatment vs Palbociclib or MRTX1133 monotherapy after 24 h of treatment; hallmarks (h.all.v2023.2). E Normalised counts of E2F targets upon 24 h treatment. F Expression of the E2F target gene CCNE1 in AsPC-1 cells treated with 5 µM Palbociclib, 0.5 µM MRTX1133, or the combination, for 24 h, and drug withdrawal for 24 or 48 h. mRNA levels were normalized to 36B4 mRNA, mean ± SD. Statistical analyses: E , F one-way ANOVA followed by Tukey’s multiple comparison; ns not significant, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. Complete statistics in Supplementary Fig. .

Article Snippet: Vehicle-control, Palbociclib (25 mg/kg, PZO199, Sigma-Aldrich) and/or MRTX1133 (30 mg/kg, HY-134813, Medchem Express) were administered i.p. once a day within weekly cycles of three consecutive treatment days followed by 4 days recovery until humane endpoint.

Techniques: Expressing, Comparison

Journal: Cell Death & Disease

Article Title: Simultaneous targeting of KRAS and CDK4 synergistically induces durable growth arrest in pancreatic cancer cells

doi: 10.1038/s41419-025-08362-w

Figure Lengend Snippet: A Proliferation of MIA PaCa-2 cells measured by automated transmission microscopy (Celigo®). Cells were reverse transfected by siRNAs to RB1 ( A ), RBL1 ( B ), RBL2 ( C ), E2F4 ( D ), or FOS ( E ); (scrb = ctrl siRNA). On day 1 after transfection, the cells were treated with DMSO, 10 µM (5 µM for ( E )) Palbociclib, 5 µM Sotorasib or the combination, for 48 h, followed by 7 days of recovery in normal medium. Means of three technical replicates ±SD. F MIA PaCa-2 cells were transfected by siRNAs to deplete CDKN1B ( F – H ) or control siRNA (scrb) during seeding. On day 1, the cells were treated with 1, 2.5, or 5 µM Palbociclib, with or without 5 µM Sotorasib, 48 h, followed by seven days of recovery without drugs. Three technical replicates, means ± SD. I Proliferation of 8661 cells (murine PDAC) wild type (WT) or CDKN1B knock-out (KO) lines (n = 3 clones, 3 technical replicates each). All cells were treated and observed as in ( A ), with the following specifics: 1 µM Palbociclib, 0.1 µM MRTX1133 or the combination. J Cell viability of 8661 WT and CDKN1B KO cells evaluated at D3 and D7 corresponding to ( I ). Statistical analyses: A – J unpaired t-test ( A – I of AUC); ns not significant, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. Complete statistics in Supplementary Fig. .

Article Snippet: Vehicle-control, Palbociclib (25 mg/kg, PZO199, Sigma-Aldrich) and/or MRTX1133 (30 mg/kg, HY-134813, Medchem Express) were administered i.p. once a day within weekly cycles of three consecutive treatment days followed by 4 days recovery until humane endpoint.

Techniques: Transmission Assay, Microscopy, Transfection, Control, Knock-Out, Clone Assay

Journal: Cell Death & Disease

Article Title: Simultaneous targeting of KRAS and CDK4 synergistically induces durable growth arrest in pancreatic cancer cells

doi: 10.1038/s41419-025-08362-w

Figure Lengend Snippet: A In vivo treatment scheme. C57BL/6J mice were orthotopically transplanted with 1.5 × 10 5 KPC cells. After tumor detection via sonography, mice were injected i.p. with (1) vehicle control, (2) Palbociclib 25 mg/kg, (3) MRTX1133 30 mg/kg and (4) Palbociclib + MRTX1133 on three consecutive days followed by a 4-days treatment break until humane endpoint. B Kaplan–Meier plot comparing overall survival since treatment initiation of the differently treated groups including the median survival. C Comparison of tumor to body weight ratio between the different treatment groups. D Representative images of H&E-stained tumor slices across the four treatment groups. Scale bar 50 µm. E Quantification of CD3 ( E ), CD4 ( F ), and CD8 ( G ) positive cells per tumor tissue. Statistical analyses: B Log-rank (Mantel-Cox) test followed by Bonferroni correction for multiple comparison was used for Kaplan–Meier survival plots; C , E , F , G one-way ANOVA followed by Tukey’s multiple comparison; ns not significant, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001.

Article Snippet: Vehicle-control, Palbociclib (25 mg/kg, PZO199, Sigma-Aldrich) and/or MRTX1133 (30 mg/kg, HY-134813, Medchem Express) were administered i.p. once a day within weekly cycles of three consecutive treatment days followed by 4 days recovery until humane endpoint.

Techniques: In Vivo, Injection, Control, Comparison, Staining

Journal: Cell Death & Disease

Article Title: Simultaneous targeting of KRAS and CDK4 synergistically induces durable growth arrest in pancreatic cancer cells

doi: 10.1038/s41419-025-08362-w

Figure Lengend Snippet: A tSNE visualization of clusters derived from single cell RNA sequencing of the mouse tumors (vehicle, Palbociclib, MRTX1133, or combination). n = 2 animals per treatment group. Different colors represent different clusters with cell numbers per cluster mentioned in brackets. Annotated cell types were identified using SingleCellNet and the expression of key markers. B Distribution of the cells of each sample in the different clusters. The numbers represent the percentage of cells per sample in each cluster, while the colors of the rows represent the identity of the cells within the cluster. C Analysis of gene ontology of differentially expressed genes of Palbociclib, MRTX1133 or combination vs vehicle in tumor cells (clusters 0–6 and 8) via metascape (downregulated, top 1000 genes). Only genes with |log2fold| ≥ 0.25 and adjusted p-value (padj.) < 0.05 were included in the analysis. Supplementary Table contains DE genes of all 15 clusters and of tumor clusters 0–6 and 8 separately. D Analysis performed as in ( C ) for the comparison of MRTX1133 vs combination (1980 genes). E Dot plot of average expression and percent expressed for tumor cells (clusters 0–6 and 8) for specific genes of the four treatment groups.

Article Snippet: Vehicle-control, Palbociclib (25 mg/kg, PZO199, Sigma-Aldrich) and/or MRTX1133 (30 mg/kg, HY-134813, Medchem Express) were administered i.p. once a day within weekly cycles of three consecutive treatment days followed by 4 days recovery until humane endpoint.

Techniques: Derivative Assay, Single Cell, RNA Sequencing, Expressing, Comparison

Journal: iScience

Article Title: Intratumor heterogeneity in KRAS signaling shapes treatment resistance

doi: 10.1016/j.isci.2024.111662

Figure Lengend Snippet:

Article Snippet: MRTX1133 , ChemieTek , Cat# CT-MTX1133.

Techniques: Virus, Recombinant, Staining, Whole Genome Amplification, Plasmid Preparation, Software, Sequencing, RNA Sequencing, Knock-Out

Journal: Cancer Discovery

Article Title: Preclinical Characterization and Clinical Activity of RNK08954 , a Highly Selective and Orally Bioavailable KRAS G12D Inhibitor

doi: 10.1158/2159-8290.CD-25-1346

Figure Lengend Snippet: RNK08954 is a potent and selective noncovalent KRAS G12D inhibitor. A, Structure of RNK08954 . B, KD values of RNK08954 for KRAS G12D , KRAS G12C , KRAS G12V , and KRAS WT were determined using inactive/active KRAS (GDP/GCP-loaded) SPR assays. C and D, Western blot assay of KRAS pathway targets p-ERK1/2 in SW1990 cells treated for 6/24/48/72 hours with RNK08954 (seven doses in total). Data are representative of several independent similar experiments; the statistics are shown in D . E, Cytotoxic effects of RNK08954 and MRTX1133 in cancer cell lines with KRAS G12D/C/V/S mutation and cell lines without KRAS mutation, also Ba/F3 cells with the expression of WT HRAS/NRAS/KRAS and corresponding G12D mutation in HRAS/NRAS/KRAS protein. Cell lines were treated with the indicated concentrations of RNK08954 and MRTX1133 for 72 hours. Cell viability was then assessed using CTG. Top concentration: 333.33 mmol/L for human tumor cell lines and 10 μmol/L for Ba/F3 cell lines. F, Scatter plot shows the IC 50 values of RNK08954 and MRTX1133 in KRAS-mutant cell lines.

Article Snippet: MRTX1133 is the first noncovalent, potent, and selective KRAS G12D inhibitor ( , ) although recently, Bristol Myers Squibb has announced termination of the development of MRTX1133 because of the high instability of the PK profile.

Techniques: Western Blot, Mutagenesis, Expressing, Concentration Assay

Journal: bioRxiv

Article Title: MRTX1133 is a potent non-covalent KRAS (G12C) inhibitor with tissue-specific activity

doi: 10.1101/2025.09.09.674973

Figure Lengend Snippet: The indicated human pancreatic ductal adenocarcinoma (PDAC) cell lines with KRAS (G12X) mutations ( A ) and isogenic and Rasless MEFs expressing human KRAS (G12X) or BRAF (V600E) mutations ( B ) were treated with MRTX1133 for 72 hours and cell viability was measured using CyQUANT ™ assays. IC 50 values were determined from curve fitting and are representative of two or three independent experiments (see Figures S1A-C for repeat experiments) with six different wells per concentration per experiment (n=6). Error bars represent standard deviation (SD). The tables in A indicate gene variants in each PDAC cell line, with full and half green-shaded boxes indicating homozygous and heterozygous alterations, respectively. Cell seeding densities: 2,000 cells/well for AsPc-1, SW1990, PaTu 8902, PSN-1; 4,000 cells/well for MIA PaCa-2, BxPc-3; 3, 000 cells/well for all Rasless MEFs.

Article Snippet: MIA PaCa-2 cells were plated in 6-cm dish (GenClone 25-202) until about 40-60% confluency and treated with fresh media containing MRTX1133 or DMSO (Fisher Bioreagents BP231-100) for a specified period (3, 6, or 24 hrs).

Techniques: Expressing, CyQUANT Assay, Concentration Assay, Standard Deviation

Journal: bioRxiv

Article Title: MRTX1133 is a potent non-covalent KRAS (G12C) inhibitor with tissue-specific activity

doi: 10.1101/2025.09.09.674973

Figure Lengend Snippet: ( A ) MRTX849, a KRAS (G12C) covalent inhibitor, was used as starting point for medicinal chemistry optimization to obtain MRTX1133, a non-covalent KRAS (G12D) inhibitor . Red highlights show acrylamide and bicyclic piperazinyl nitrogen warheads for G12C and G12D mutant specificity, respectively. ( B ) Molecular docking of MRTX1133 on GDP-bound KRAS (G12C) (PDB 6UT0) predicts strong binding affinity (Vina docking score -13 kcal/mol) and a similar binding pose to MRTX849 including shared H-bonding interactions (yellow dashed lines). ( C ) Microscale thermophoresis and calculated K D for the interaction of recombinant GDP-KRAS (G12C) with MRTX1133 using three different readings from one capillary tube per concentration (n=3) (see Methods). Error bars = SD.

Article Snippet: MIA PaCa-2 cells were plated in 6-cm dish (GenClone 25-202) until about 40-60% confluency and treated with fresh media containing MRTX1133 or DMSO (Fisher Bioreagents BP231-100) for a specified period (3, 6, or 24 hrs).

Techniques: Mutagenesis, Binding Assay, Microscale Thermophoresis, Recombinant, Concentration Assay

Journal: bioRxiv

Article Title: MRTX1133 is a potent non-covalent KRAS (G12C) inhibitor with tissue-specific activity

doi: 10.1101/2025.09.09.674973

Figure Lengend Snippet: (A) Human lung cancer cell lines with KRAS (G12C) mutation were treated with MRTX1133 for 72 hours and cell viability was measured using CyQUANT ™ assays. IC 50 values were determined from curve fitting and are representative of two independent experiments (see Figure S2A for a repeat experiment) with six different wells per concentration per experiment (n=6). Cells were seeded at 2,000 cells/well. Error bars = SD. Shaded boxes in the respective tables indicate gene variants per cancer cell line (full – homozygous, half – heterozygous). (B) 3D spheroids of the same lung cancer cell lines treated with MRTX1133 for 3 days and cell viability was measured using CellTiter-Glo reagent. IC 50 values were determined from curve fitting using three different wells per concentration seeded from one cell culture flask (n=3). Cells were seeded at 5, 000 cells/well. Error bars = SD. (C) The same as in B but for the MIA PaCa-2 cell line treated with MRTX1133 or adagrasib. IC 50 curves are representative of two independent experiments (see Figure S2B for repeat experiments) with three different wells per concentration per experiment. (D) The lung cancer cell lines were treated with 100 nM or 1,000 nM of MRTX1133 for 6 hr or 24 hr and lysates were immunoblotted for pERK and total ERK with vinculin as loading control. The blots shown here are representatives of two independent experiments for each cell line.

Article Snippet: MIA PaCa-2 cells were plated in 6-cm dish (GenClone 25-202) until about 40-60% confluency and treated with fresh media containing MRTX1133 or DMSO (Fisher Bioreagents BP231-100) for a specified period (3, 6, or 24 hrs).

Techniques: Mutagenesis, CyQUANT Assay, Concentration Assay, Cell Culture, Control

Journal: bioRxiv

Article Title: MRTX1133 is a potent non-covalent KRAS (G12C) inhibitor with tissue-specific activity

doi: 10.1101/2025.09.09.674973

Figure Lengend Snippet: MIA PaCa-2 cells were treated with indicated concentrations of MRTX1133 (A) or 100 nM of MRTX1133, adagrasib and sotorasib ( B and C ) for 3, 6, 24 hours and cell lysates were harvested and immunoblotted for pERK and total ERK. Shown in panels A and B are representative western blots from two independent experiments (see Figures S3A, B for repeat experiments), and changes in pERK levels shown in C were quantified relative to total ERK levels and normalized to untreated (DMSO) samples. *p<0.05 on ordinary one-way ANOVA. n.s. = not significant. MRTX1133, adagrasib, and sotorasib have significantly lower values (p<0.05) than DMSO control in all time points (asterisks not shown).

Article Snippet: MIA PaCa-2 cells were plated in 6-cm dish (GenClone 25-202) until about 40-60% confluency and treated with fresh media containing MRTX1133 or DMSO (Fisher Bioreagents BP231-100) for a specified period (3, 6, or 24 hrs).

Techniques: Western Blot, Control

Journal: bioRxiv

Article Title: MRTX1133 is a potent non-covalent KRAS (G12C) inhibitor with tissue-specific activity

doi: 10.1101/2025.09.09.674973

Figure Lengend Snippet: ( A ) Mice bearing MIA PaCa-2 xenografts were treated twice a day (BID) by intraperitoneal administration of 30 mg/kg MRTX1133 (n=8) or vehicle (n=8) for 28 days. ( B ) Mice were weighed periodically during the treatment duration. ( C ) Tumors were harvested and weighed on the 29 th day. ( D ) Harvested tumors. Error bars in A-C represent standard error of the mean (SEM); *p<0.05 on two-tailed, unpaired t-test.

Article Snippet: MIA PaCa-2 cells were plated in 6-cm dish (GenClone 25-202) until about 40-60% confluency and treated with fresh media containing MRTX1133 or DMSO (Fisher Bioreagents BP231-100) for a specified period (3, 6, or 24 hrs).

Techniques: Two Tailed Test

Journal: Communications Chemistry

Article Title: Optimizing drug design by merging generative AI with a physics-based active learning framework

doi: 10.1038/s42004-025-01635-7

Figure Lengend Snippet: A Scatter plot of KRAS G12D unknown molecules with a Glide gscore below −8 kcal·mol −1 and a maximum similarity to the initial-specific set below 0.30. Red dotted lines indicate the stringent thresholds of −9 kcal·mol −1 for gscore and 0.25 for similarity used during candidate selection. B Linear regression and correlation coefficient between Glide gscore and ABFE ΔG (left) and between PELE BFE and ABFE ΔG (right) for the 19 candidate drugs selected after PELE rescoring. C Table showing MM affinity predictions and maximum similarity to the unknown initial-specific set molecules for molecules with potential activity against KRAS. D Structures of molecules with potential activity against KRAS by ABFE simulations. E MRTX1133 Mirati’s KRAS G12D inhibitor currently in clinical trials.

Article Snippet: To perform the HTVS, we used a grid created with the Mirati MRTX1133 molecule and two constraints: (1) a distance constraint between the molecule's basic amine and G12D, (2) and a hydrophobic aromatic constraint with the hydrophobic subcavity of SII.

Techniques: Selection, Activity Assay, Clinical Proteomics

Journal: EMBO Reports

Article Title: FOXO1 links KRAS G12D and G12V alleles to glutamine and nitrogen metabolism in colorectal cancer

doi: 10.1038/s44319-025-00641-z

Figure Lengend Snippet: ( A ) Representative immunoblot showing increased expression of FOXO1 in G12D and G12V mutant SW48 cells in full media. Graph is a means ± standard deviations of three biological replicates. Statistical analyses were performed using one-way ANOVA followed by Dunnett’s multiple comparisons test comparing all mutants to the WT. Only statistically significant p values are shown (G12D ** p = 0.009, G12V * p = 0.024). ( B ) mRNA levels of FOXO1 detected by qPCR. Data shows means ± standard deviations log2FC change of 3 biological repeats in full media. Statistical analyses were performed using one-way ANOVA followed by Dunnett’s multiple comparisons test comparing all mutants to the WT. Only statistically significant p values are shown (G12D * p = 0.03, G12V * p = 0.014). ( C , D ) Representative immunoblots showing the increased expression of FOXO1 in G12D and G12V mutant SW48 cells in full media ( C ) or low nutrient conditions ( D ) with MRTX1133 (100 nM) compared to matched DMSO controls. MRTX1133 decreases the expression of AKAP12, FOXO1 only in SW48 +/G12D cells. GS expression is altered but only in low-nutrient media. Red asterisks mark the relevant G12D lanes. Quantifications of western blots are shown as means ± standard deviations of three biological replicates. Statistical analyses were performed using one-way ANOVA followed by Dunnett’s multiple comparisons test for all comparisons. Only statistically significant p values are shown ( C AKAP12 **** p ≤ 0.0001; FOXO1 * p = 0.012, *** p = 0.0004, G12D * p = 0.012; GS **** p ≤ 0.0001, *** p = 0.0004; D AKAP12 G12C * p = 0.0279, **** p ≤ 0.0001, G12V * p = 0.014, *** p = 0.0001; FOXO1 G12C ** p = 0.004, *** p = 0.0001, **** p ≤ 0.0001, G12D ** p = 0.004; GS **** p ≤ 0.0001, *** p = 0.0001). ( E , F ) Representative immunostaining of FOXO1 (red) and nuclear staining (blue) in SW48 isogenic panel in low-nutrient conditions. Scale bar: 25 µm; inserts: 3x magnification. ( F ) Quantification of mean nuclear intensity (related to E ) and standard errors (cells per sample ≥500, n = 3 biological repeats). Statistical analyses were performed using one-way ANOVA followed by Dunnett’s multiple comparisons test comparing all mutants to the WT. Only statistically significant p values are shown (**** p ≤ 0.0001). ( G ) qPCR analysis of GLUL mRNA levels upon FOXO1 inhibition in full media. Values were normalised to SW48 WT DMSO of each experiment and shown as mean with standard errors. Statistical analyses from 3 biological replicates were performed using two-way ANOVA followed by Sidak’s multiple comparisons test for all comparisons. Only statistically significant p values are shown (* p = 0.0424, *** p = 0.0002, **** p ≤ 0.0001). ( H ) Changes in abundance of glutamine ( m + 2) upon inhibition of FOXO1 in a 13 C-glucose labelling experiment (low nutrient). Data represents the mean of 5 technical repeats and standard deviations. Statistical analyses were performed using two-way ANOVA followed by Sidak’s multiple comparisons test for all comparisons. Only statistically significant p values are shown (*** p = 0.0001, **** p ≤ 0.0001). .

Article Snippet: MRTX1133 , FisherScientific , Cat#18754856.

Techniques: Western Blot, Expressing, Mutagenesis, Immunostaining, Staining, Inhibition