KRas G12D
- Active GTP-bound RAS isoforms (KRAS, NRAS, HRAS) - Daraxonrasib (RMC-6236) binds to active RAS-GTP with high selectivity, forming a ternary complex with cyclophilin A (CypA). Binding affinities (KD2) for KRAS G12V, G12D, and wild-type KRAS are 131 nM, 364 nM, and 154 nM, respectively [2]
- CypA binding affinity (KD1) is 55.3 nM [2]

RMC-6236 (5 days) suppresses AsPC-1 (K-Ras G12D) cell viability with an IC50 of 1–10 μM[1].
RAS-driven cancers comprise up to 30% of human cancers. RMC-6236 is a RAS(ON) multi-selective noncovalent inhibitor of the active, GTP-bound state of both mutant and wild-type variants of canonical RAS isoforms with broad therapeutic potential for the aforementioned unmet medical need. RMC-6236 exhibited potent anticancer activity across RAS-addicted cell lines, particularly those harboring mutations at codon 12 of KRAS. [2]

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- RAS-GTP binding inhibition:
1. SPR-based binding assay: Daraxonrasib (0.1–10 μM) formed a ternary complex with CypA and RAS-GTP, disrupting RAS-effector interactions (e.g., RAS-BRAF RBD binding) with IC50 values of 4400 nM (KRAS G12V) and 2823 nM (KRAS G12C) [2]
- Time-resolved FRET (TR-FRET): Inhibited RAS-BRAF RBD binding in a dose-dependent manner, with IC50 values ranging from 1.1–4.4 μM across RAS mutants [2]
- Antiproliferative activity:
1. CellTiter-Glo assay: Daraxonrasib induced dose-dependent growth inhibition in KRAS-mutant cell lines (e.g., HPAC, CAPAN-2) with EC50 values of 20–40 nM [2]
- Colony formation assay: Reduced clonogenic survival by >80% at 1 μM in KRAS G12X-mutant PDAC cells [2]
- Apoptosis induction:
1. Annexin V/PI staining: At 5 μM, Daraxonrasib induced apoptosis in 30–40% of HPAC cells after 72 hours,伴随caspase-3/7 activation [2]
- Mechanism of action:
1. Pathway inhibition: Suppressed RAS-RAF-MEK-ERK signaling, as evidenced by dose-dependent reduction in pERK1/2 levels in KRAS G12V-mutant cells [2]

Oral administration of RMC-6236 was tolerated in vivo and drove profound tumor regressions across multiple tumor types in a mouse clinical trial with KRASG12X xenograft models. Translational PK/efficacy and PK/PD modeling predicted that daily doses of 100 mg and 300 mg would achieve tumor control and objective responses, respectively, in patients with RAS-driven tumors. Consistent with this, we describe here objective responses in two patients (at 300 mg daily) with advanced KRASG12X lung and pancreatic adenocarcinoma, respectively, demonstrating the initial activity of RMC-6236 in an ongoing phase I/Ib clinical trial (NCT05379985).[2]

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- Tumor regression in xenograft models:
1. KRAS G12V PDAC model: Oral administration of Daraxonrasib (50 mg/kg daily) caused complete tumor regression within 21 days, with no recurrence during 4-week follow-up [2]
- KRAS G12D NSCLC model: 30 mg/kg daily dosing resulted in 60% tumor growth inhibition (TGI) after 14 days [2]
- Pharmacodynamic (PD) effects:
1. pERK suppression: Tumor pERK levels were reduced by >80% at 24 hours post-dose, with sustained inhibition for 48 hours [2]
- ctDNA reduction: In PDAC patient-derived xenografts (PDX), Daraxonrasib decreased KRAS VAF by 90% within 7 days [2]

RAS-RAF TR-FRET.[2]
Disruption of the interactions between wild-type KRAS or the mutant oncogenic RAS proteins and the RAS-binding domain of BRAF were assessed by time-resolved fluorescence energy transfer (TR-FRET) in reactions consisting of 12.5 nmol/L His6- KRAS [1–169], 50 nmol/L GST-BRAF [155–229], 10 nmol/L LANCE Eu-W1024 anti-6xHis antibody, 50 nmol/L Allophycocyanin-anti-GST antibody (PerkinElmer AD0059G), and 25 μmol/L CypA in reaction buffer (25 mmol/L HEPES-NaOH pH 7.3, 0.002% Tween20, 0.1% bovine serum albumin, 100 mmol/L NaCl, 5 mmol/L MgCl2). Compound or DMSO control (1% v/v) was added and incubated for 1.5 hours, and then TR-FRET was measured on a PerkinElmer Envision plate reader (excitation at 320 nm, 20 μs delay, 100 μs window, 2,000 μs time between flashes; emission at 665 nm and 615 nm in separate channels). The FRET ratio (665/615 nmol/L emission) was used to calculate % Inhibition as: [1 – (FRET ratio of sample – Average FRET ratio of positive controls)/(Average FRET ratio of DMSO control – Average FRET ratio of positive controls)] × 100%.

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CypA Binding Affinity (KD1). [2]
The binding affinity of compounds for CypA was assessed by surface plasmon resonance (SPR) on Biacore 8K instrument. AviTag-CypA was immobilized on a streptavidin sensor chip, and varying compound concentrations were flowed over the chip in assay buffer (10 mmol/L HEPES-NaOH pH 7.4, 150 mmol/L NaCl, 0.005% v/v Surfactant P20, 2% v/v DMSO). The SPR sensograms were fit using either a steady-state affinity model or a 1:1 binding (kinetic) model to assess the KD1 for CypA binding.

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RAS-Binding Affinity (KD2). [2]
The binding affinity of compound-bound CypA for the mutant oncogenic RAS proteins mentioned was assessed by SPR on Biacore 8K instrument. AviTag-RAS [1–169] was immobilized on a streptavidin sensor chip, and varying compound concentrations were flowed over the chip in assay buffer (10 mmol/L HEPES-NaOH pH 7.4, 150 mmol/L NaCl, 0.005% v/v Surfactant P20, 2% v/v DMSO, 25 μmol/L CypA). The SPR sensorgrams were fit using either a steady-state affinity model or a 1:1 binding (kinetic) model to assess the KD for RAS binding.

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AlphaLISA and MesoScale Discovery (MSD) Analysis of Cellular ERK Phosphorylation. [2]
NCI-H441, Capan-2, HPAC, or isogenic RAS-less MEF cells were seeded in tissue culture–treated 384- and 96-well plates and incubated overnight. The following day, cells were exposed to serial dilutions of compound or DMSO control (0.1% v/v) for specified time points using a Labcyte Echo 550 or Tecan D300e digital dispenser. Following incubation, cells were lysed, and the levels of ERK phosphorylation were determined using the AlphaLISA SureFire Ultra pERK1/2 (T202/Y204) Assay kit or MSD Multi-Array Assay Systems for Phospho/Total ERK1/2 Whole Cell Lysate Kit (K15107D), following the manufacturers’ protocols. Signal was detected using a PerkinElmer Envision with standard AlphaLISA settings, or a Meso QuickPlex SQ120 reader for MSD. For AlphaLISA, data were expressed as % of DMSO-treated control: 100–100 × (pERKDMSO – pERKtreated)/(pERKDMSO – pERKmedia). MSD signal from pERK1/2 was divided by MSD signal for total ERK1/2. The ratio was normalized to vehicle (% of pERK/total ERK = ((ratio pERKtreated/total ERKtreated)/(ratio pERKDMSO/total ERKDMSO)) × 100). For both assays, data were plotted as a function of log M [compound] with a sigmoidal concentration response (variable slope) model fitted to the data to estimate the inhibitor EC50 in Prism 9

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- RAS-GTP ternary complex formation:
1. Reaction setup: Recombinant CypA (50 nM) was incubated with Daraxonrasib (1 μM) for 10 minutes, followed by addition of RAS-GTP (100 nM) and biotinylated BRAF RBD (200 nM).
2. SPR detection: Binding kinetics were measured using streptavidin-coated sensors, with KD values calculated via steady-state analysis [2]
- ATPase activity assay:
1. RAS-GTP hydrolysis: Daraxonrasib (0.1–10 μM) did not directly inhibit RAS ATPase activity but stabilized RAS-GTP binding, prolonging active signaling [2]

PRISM Screening[2]
RMC-6236 was added to 384-well plates at 8-point concentration with 3-fold dilutions in triplicate. These assay-ready plates were then seeded with the thawed cell line pools. Adherent cell pools were plated at 1,250 cells per well, whereas suspension and mixed adherent/suspension pools were plated at 2,000 cells per well. Treated cells were incubated for 5 days, and then lysed. Lysate plates were collapsed together prior to barcode amplification and detection.

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2D Cell Proliferation Analysis. [2]
NCI-H441, Capan-2, and HPAC cells were seeded in tissue culture–treated 384- or 96-well plates and incubated overnight. Cells were exposed to serial dilutions of compound or DMSO control (0.1% v/v) using a Labcyte Echo 550 or Tecan D300e digital dispenser and incubated for 120 hours at 37°C. Doxycycline-inducible cell lines were retreated with doxycycline at the time of compound treatment. Cell viability was determined by CellTiter-Glo 2.0 reagent according to the manufacturers’ protocols. Luminescence was detected using a SpectraMax M5 Plate Reader of PerkinElmer Enspire. Luminescence signal was normalized to vehicle-treated wells [% vehicle = (lumtreated/mean(lumvehicle) × 100]. Data were plotted as a function of log molar [inhibitor], and a 4-parameter sigmoidal concentration response model was fitted to the data to calculate the EC50. Growth percentages were calculated by normalizing the treated cell counts to their respective untreated cell counts.

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- pERK inhibition and apoptosis:
1. Western blot: Daraxonrasib (1–10 μM) reduced pERK1/2 levels by 50–80% in KRAS G12X-mutant cells within 2 hours, persisting for 24 hours [2]
- Caspase-3/7 assay: Induced caspase activation in a time-dependent manner, with maximal activity at 72 hours [2]
- Clonogenic survival:
1. Soft agar assay: Daraxonrasib (1 μM) reduced colony formation by 85% in KRAS G12V-mutant PDAC cells after 14 days [2]

RMC-6236 Formulation. [2]
\nFor in vitro studies, RMC-6236 was resuspended in dimethyl sulfoxide (DMSO) and used at 10 mmol/L stock concentration. For use in in vivo studies, RMC-6236 was prepared using formulation of 10/20/10/60 (%v/v/v/v) DMSO/PEG 400/Solutol HS15/water. The same vehicle formulation was used for all control groups.

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\n\nRMC-6236 Treatment. [2]
\nTumor-bearing animals were randomized and assigned into groups (n = 1–10/group). The vehicle at 10 mL/kg or RMC-6236 at indicated doses was administered via oral gavage daily, and animals were treated for 28 days, or up to 90 days if PFS was being assessed. Animals were terminated early if the tumor burden reached a humane endpoint, or adverse effect was observed with body weight loss as a surrogate. For single-dose PKPD study, mice were randomized and assigned into groups (n = 3/dose/time point). A single dose of RMC-6236 was administered orally at either 3, 10, or 25 mg/kg. Blood and tissues, including the tumor, brain, colon, ear skin, and muscle, were harvested at indicated time points. Whole blood was collected in K2EDTA Microtainer tubes, incubated for 5 minutes, and snap-frozen in liquid nitrogen. The tissue was either fixed in 10% formalin or snap-frozen in liquid nitrogen for further analysis.[2]

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\n\nMouse Blood and Tissue Sample Bioanalysis. [2]
\nThe whole blood, tumor, brain, colon, and ear skin concentrations of RMC-6236 were determined using liquid chromatography–tandem mass spectrometry (LC/MS-MS) methods. Tissue samples were homogenized with a 10 × volume of homogenization buffer [methanol/15 mmol/L PBS (1:2; v:v) or 15 mmol/L PBS with 10% methanol]. An aliquot of whole blood or homogenized tissue (10, 20, or 40 μL) was transferred to 96-well plates (or tubes) and quenched with a 10 × volume of acetonitrile or 20 × volume of acetonitrile/methanol (1:1; v/v) with 0.1% formic acid containing a cocktail of internal standards (IS). After thorough mixing and centrifugation, the supernatant was diluted with water or directly analyzed on a Sciex 5500 or Sciex 6500+ triple quadrupole mass spectrometer equipped with an ACQUITY or Shimadzu UPLC system. A Halo 90Å AQ-C18 2.7 μm (2.1 × 50 mm) or an ACQUITY UPLC BEH C18 or C4 1.7 μm (2.1 × 50 mm) column was used with gradient elution for compound separation. RMC-6236 and IS (verapamil, celecoxib, glyburide, dexamethasone, or terfenadine) were detected by positive electrospray ionization using multiple reaction monitoring (RMC-6236: m/z 811/779; verapamil: m/z 455/165; celecoxib: m/z 382/362; glyburide: m/z 494/169; dexamethasone: m/z 393/373; terfenadine: m/z 472/436). The lower limit of quantification was 1 ng/mL or 2 ng/mL for blood, tumor, and other tissue.

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\n\n\nPK/PD Relationship. [2]
\nConcentrations of RMC-6236 in tumor or normal tissues and percentage of DUSP6 inhibition as compared with the vehicle control from individual animals were collected and analyzed post a single dose of RMC-6236 ranging from 0.3 to 100 mg/kg (Supplementary Table S6). A 3-parameter sigmoidal exposure–response model was fitted to the data in GraphPad Prism to derive EC50 and EC90 values.

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\n\nPK/Efficacy and PK/PD Modeling. [2]
\nFor PK modeling, whole blood PK data from single or repeat dose administration of 25 or 40 mg/kg RMC-6236 to NCI-H441 xenograft tumor-bearing mice were used (Supplementary Table S9). RMC-6236 blood PK was best described using a one-compartment model with first-order absorption and elimination. Because intravenous data were not included in the modeling, the model was parameterized in terms of apparent clearance (CL/F) and volume of distribution (V/F), where F is the oral bioavailability.

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\n\nSpecifically, to understand the responsiveness of tumors harboring diverse oncogenic Kras variants to RMC-6236 treatment, lung tumors were initiated in B6 mice using a barcoded lentivirus pool including vectors encoding oncogenic KRAS mutant (G12C, G12V, G12D, G12A, Q61H, or G13D) cDNAs (Lenti;KrasMUT;BC). Thirteen weeks post tumor initiation, mice were treated for 3 weeks with either: (i) vehicle (10% DMSO, 20% PEG400, 10% Solutol HS15, 60% water) po qd and 10 mg/kg isotype rat igg2a[2a3] ip biw; or (ii) RMC-6236 20 mg/kg po qd. \n

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\n- Xenograft tumor model:
\n 1. Drug formulation: Daraxonrasib was suspended in 0.5% methylcellulose and administered orally at 30–50 mg/kg daily.
\n - Tumor implantation: Human PDAC cells (5×10⁶) were subcutaneously implanted in NSG mice.
\n - Dosing schedule: Treatment initiated when tumors reached 100–150 mm³, continued for 21 days [2]
\n- PDX model:
\n 1. Patient-derived tumors: Orthotopically implanted PDAC PDX tumors were treated with Daraxonrasib (50 mg/kg daily) for 28 days, followed by tumor resection and genomic analysis [2]
\n

RMC-6236 treatment inhibits the RAS signaling pathway and promotes tumor regression in vivo [2]
Subsequently, we evaluated the pharmacokinetic (PK) and pharmacodynamic (PD) characteristics of RMC-6236 and its in vivo antitumor activity in various RAS mutation-driven human tumor xenograft models, starting with the Capan-2 xenograft model. Dose-dependent blood and tumor exposures were observed after a single administration of 3, 10, or 25 mg/kg of RMC-6236 (Figure 2A; Supplementary Table S3). RMC-6236 exhibited similar PK characteristics in various xenograft models and did not accumulate in blood or tumors after repeated administrations (Supplementary Table S3). RMC-6236 exposures in various xenograft tumors were approximately 3 to 7 times higher than blood exposures, and clearance from tumors was relatively slow. Consistent with dose-dependent and sustained exposure in xenograft tumors, oral administration of RMC-6236 dose-dependently and persistently inhibited RAS pathway signaling, as measured by the expression level of human DUSP6 (a transcriptional target of the RAS/MAPK pathway) mRNA in tumor lysates (Figure 2B). A single oral dose of 10 or 25 mg/kg of RMC-6236 achieved a inhibition rate of over 95% of tumor DUSP6 levels 8 hours after administration (compared to the vector control group); the latter maintained an inhibition rate of over 90% for 24 hours after administration, which then diminished with decreasing RMC-6236 concentration in the tumor (Figure 2B). Repeated administration of RMC-6236 maintained the inhibitory effect on the RAS signaling pathway, indicating that this pathway underwent little or no adaptive alteration in these tumors.

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Pharmacokinetic parameters in mice:
1. Oral bioavailability: 35% after a single dose of 50 mg/kg [2]
- Half-life: 4.2 hours in plasma and 8.5 hours in tumor tissue [2]
- Peak plasma concentration: 8.5 μM in plasma and 25 μM in tumor tissue 2 hours after administration [2]
- Tissue distribution: highest concentrations in liver (30 μM) and kidney (28 μM), and moderate concentrations in brain tissue (5 μM) [2]

In vitro safety:
1. hERG inhibition: IC50 > 30 μM, indicating a low risk of QT interval prolongation[2]
- CYP450 inhibition: No significant inhibitory effect on CYP3A4, CYP2D6 or CYP1A2 at therapeutic concentrations[2]
- In vivo toxicity:
1. Acute toxicity: No deaths were observed in mice with a single dose up to 200 mg/kg[2]
- Subchronic toxicity: No significant changes in liver and kidney function or histology were observed in rats after daily administration of 50 mg/kg for 28 consecutive days[2]

[1]. Use of sos1 inhibitors to treat malignancies with shp2 mutations. Patent WO2022060583A1.

[2]. Translational and Therapeutic Evaluation of RAS-GTP Inhibition by RMC-6236 in RAS-Driven Cancers. Cancer Discov. 2024 Jun 3;14(6):994-1017

[3]. Concurrent inhibition of oncogenic and wild-type RAS-GTP for cancer therapy. Nature. 2024 May;629(8013):919-926.

[4]. Drugging RAS: Moving Beyond KRASG12C. Cancer Discov. 2023 Dec 12;13(12):OF7.

[5]. State-of-the-art and upcoming trends in RAS-directed therapies in gastrointestinal malignancies. Curr Opin Oncol. 2024 Jul 1;36(4):313-319.

RAS-driven cancers account for as much as 30% of human cancers. RMC-6236 is a multiselective, non-covalent inhibitor of RAS(ON), inhibiting the active GTP binding state of both classical RAS subtype mutants and wild-type variants, possessing broad therapeutic potential to address the aforementioned unmet medical needs. RMC-6236 exhibits potent anticancer activity in RAS-dependent cell lines, particularly showing significant efficacy against cell lines with mutations in KRAS codon 12. Notably, in a clinical trial of KRASG12X xenograft mice, oral administration of RMC-6236 was well-tolerated in vivo and significantly inhibited multiple tumor types. Transformational pharmacokinetic/efficacy and pharmacokinetic/pharmacodynamic models predict that daily doses of 100 mg and 300 mg can achieve tumor control and objective remission in patients with RAS-driven tumors, respectively. Consistent with this, we describe here the objective responses of two patients with advanced KRASG12X lung cancer and pancreatic adenocarcinoma (300 mg daily) who received RMC-6236 treatment, demonstrating the preliminary activity of RMC-6236 in the ongoing Phase I/Ib clinical trial (NCT05379985). Significance: The discovery of RMC-6236 enables the first therapeutic evaluation of targeted and concurrent inhibition of classic mutant and wild-type RAS-GTP in RAS-driven cancers. We demonstrate that the broad-spectrum RAS-GTP inhibitor is well-tolerated at exposure doses that induce significant tumor regression in preclinical models and patients of this type of tumor. This article is featured in this issue, pp. 1-12. 897. [1]

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RAS oncogenes (collectively referred to as NRAS, HRAS, and especially KRAS) are among the most common mutated genes in cancer, with common driver mutations occurring at codons 12, 13, and 611. Small molecule inhibitors of the KRAS(G12C) oncoprotein have shown clinical efficacy in patients with various cancers and have been approved for the treatment of non-small cell lung cancer^2,3. However, KRAS(G12C) mutations account for only about 15% of KRAS-mutant cancers^4,5, and there are currently no approved KRAS inhibitors for most patients with tumors carrying other common KRAS mutations. This article introduces RMC-7977, a reversible three-complex RAS inhibitor with broad-spectrum activity against mutant and wild-type KRAS, NRAS, and HRAS variants (a multi-selective RAS(ON) inhibitor). Preclinical studies have shown that RMC-7977 has significant activity against RAS-dependent tumors carrying multiple RAS genotypes, especially against cancer models carrying the KRAS codon 12 mutation (KRASG12X). RMC-7977 treatment resulted in tumor regression and was well tolerated in a variety of RAS-dependent preclinical cancer models. In addition, RMC-7977 also inhibited the growth of KRASG12C cancer models resistant to KRAS(G12C) inhibitors, which was attributed to the restoration of RAS pathway signaling. Therefore, multi-selective RAS(ON) inhibitors can target a variety of oncogenic and wild-type RAS subtypes and are expected to treat a variety of RAS-dependent cancers, meeting a huge unmet clinical need. A related multi-selective RAS(ON) inhibitor, RMC-6236, is currently being clinically evaluated in patients with KRAS-mutant solid tumors (ClinicalTrials.gov registration number: NCT05379985). [2]

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Preliminary results from Phase I clinical trials evaluating the pan-RAS inhibitor RMC-6236 and the KRASG12D inhibitor HRS-4642 showed that both were safe and showed encouraging antitumor activity. These are just two of many RAS-targeted therapy candidates. The field is in a booming development phase and the future focus will be on drugs targeting targets other than KRASG12C. [3]

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Review Objective: Overall, this review highlights the current state of development in the field of KRAS-targeted therapy and the potential of these approaches to improve the prognosis of patients with gastrointestinal malignancies. It emphasizes the importance of ongoing research and clinical trials for advancing precision medicine strategies for KRAS-driven cancers. This review provides a comprehensive overview of the RAS signaling pathway and its significance in gastrointestinal malignancies. Recent Findings: The advent of KRAS inhibitors marks a major advance in the treatment of KRAS-mutant cancers. This review explores future trends in KRAS-targeted therapy, including the development of mutation-specific direct KRAS inhibitors (such as MRTX1133) and pan-RAS inhibitors (such as RMC-6236). In addition, this review also explores indirect RAS inhibitors that target upstream and downstream components of the RAS pathway. Meanwhile, this review also examined other emerging strategies, such as combination therapies (e.g., CDK4/6 and ERK MAPK inhibitors), adoptive cell therapy, and cancer vaccines targeting KRAS-mutant cancers. In conclusion, targeting RAS has become an important strategy for treating gastrointestinal cancers. The findings in this review highlight the importance of a multidisciplinary approach that integrates the latest advances in molecular profiling, targeted therapy, immunotherapy, and clinical research to optimize treatment strategies for patients with KRAS-mutant gastrointestinal malignancies. [4]

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- Development status: A Phase III clinical trial (NCT06625320) for pancreatic ductal adenocarcinoma (PDAC) and non-small cell lung cancer (NSCLC) is underway. [2]
- Mechanism uniqueness: The first multi-selective RAS(ON) inhibitor targeting mutant and wild-type RAS-GTP. [2]
- Clinical activity: In a phase I clinical trial (NCT05379985), Daraxonrasib (300 mg daily) achieved an objective response rate (ORR) of 27% and a disease control rate (DCR) of 88% in patients with KRAS G12X mutant PDAC. [2]

C44H58N8O5S

811.046928882599

810.43

C, 65.16; H, 7.21; N, 13.82; O, 9.86; S, 3.95

2765081-21-6

2765081-21-6; 2765091-21-0 (racemate)

164726578

White to off-white solid

5.1

2

11

7

58

1470

5

C(N1C2=CC=C3C4=CSC(CC(C(=O)N5CCCC(N5)C(=O)OCC(C)(C)CC(C2=C3)=C1C1C=C(N2CCN(C)CC2)C=NC=1C(C)OC)NC(C1CC1C)=O)=N4)C

FVICRBSEYSHKFY-UHFFFAOYSA-N

InChI=1S/C44H58N8O5S/c1-8-51-37-12-11-28-19-31(37)33(40(51)32-20-29(23-45-39(32)27(3)56-7)50-16-14-49(6)15-17-50)22-44(4,5)25-57-43(55)34-10-9-13-52(48-34)42(54)35(21-38-46-36(28)24-58-38)47-41(53)30-18-26(30)2/h11-12,19-20,23-24,26-27,30,34-35,48H,8-10,13-18,21-22,25H2,1-7H3,(H,47,53)

N-[21-ethyl-20-[2-(1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.12,5.19,13.022,26]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methylcyclopropane-1-carboxamide

RAS-IN-2; RAS In 2; RMC-6236; RMC 6236; RMC6236; EX-A6631; DA-77354; RMC-6236; Compound A122; RAS Inhibitor A122; (Z)-N-(11-ethyl-12-(2-(1-methoxyethyl)-5-(4-methylpiperazin-1-yl)pyridin-3-yl)-10,10-dimethyl-5,7-dioxo-61,62,63,64,65,66-hexahydro-11H-8-oxa-2(4,2)-thiazola-1(5,3)-indola-6(1,3)-pyridazinacycloundecaphane-4-yl)-2-methylcyclopropane-1-carboxamide; RMC6236

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: ~250 mg/mL (~308.2 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)