hMDM2 (IC50 = 1.7±0.1 nM); NVP-CGM097 specifically targets the human homolog of Mouse Double Minute 2 (HDM2 or MDM2). It binds to the p53-binding pocket on the surface of the MDM2 protein. The binding affinity (IC50) of NVP-CGM097 for MDM2 is approximately 1.7 ± 0.1 nM, demonstrating high potency. It shows significant selectivity for the p53-MDM2 interaction over other protein-protein interactions, such as p53-MDM4 (1176-fold selectivity) .
The primary target of NVP-CGM097 sulfate is the MDM2 protein, specifically the human homolog (hMDM2). MDM2 is a key negative regulator of the tumor suppressor protein p53. It binds to the N-terminal transactivation domain of p53, thereby inhibiting p53's transcriptional activity and promoting its degradation via the ubiquitin-proteasome pathway. NVP-CGM097 sulfate is a highly potent and selective inhibitor of this interaction, with an IC₅₀ of 1.7 ± 0.1 nM for hMDM2. It exhibits remarkable selectivity over MDM4 (also known as MDMX), a homolog of MDM2, with an IC₅₀ of 2000 nM, representing a >1000-fold selectivity. It is about four times more potent than Nutlin-3a, a well-known MDM2 inhibitor. By binding to MDM2, NVP-CGM097 sulfate effectively prevents the MDM2-p53 interaction, allowing p53 to accumulate and become transcriptionally active. This leads to the upregulation of p53 target genes involved in cell cycle arrest, apoptosis, and DNA repair.

In vitro, NVP-CGM097 effectively inhibits the proliferation of cancer cells harboring wild-type p53 (p53wt) in a p53-dependent manner. For instance, in p53wt neuroendocrine tumor cells (GOT1), treatment with 2,500 nM NVP-CGM097 for 96 hours resulted in a significant reduction in cell viability to 47.7%. In ER-positive breast cancer cell lines (MCF-7, ZR75-1), sub-micromolar IC50 values (approx. 0.2 μM) were observed, while p53-mutant cells (T-47D) were resistant (IC50 ~7.2 μM). Mechanistically, it induces nuclear translocation of p53, increases the expression of p53 target proteins like p21 and PUMA, and leads to G1 and G2/M cell cycle arrest followed by apoptosis .

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NVP-CGM097 sulfate binds to human MDM2 with an IC50 of 1.7 nM and demonstrates strong selectivity for MDM4 (IC50=2000 nM). NVP-CGM097 sulfate is roughly four times more powerful than Nutlin-3a (IC50=8 nM). In addition, NVP-CGM097 sulfate showed no significant action on Bcl-2:Bak, Bcl-2:Bad, Mcl-1:Bak, Mcl-1:NOXA, XIAP:BIR3 and c-IAP:BIR3 protein-protein responses. CGM097 sulfate effectively reduced the growth of cells expressing wild-type p53 while sparing p53-null cells with a 35-58-fold differential. NVP-CGM097 sulfate may dramatically relocate wild-type p53 into the nucleus, with an IC50 of 0.224 μM, indicating that it can suppress NVP-CGM097 sulfate significantly reduces the proliferation of cells expressing wild-type p53, at a 35-58-fold rate. Differential retention of p53 null cells. NVP-CGM097 sulfate inhibits HCT116 (p53WT/WT) with an IC50 of 454±136nM[1].
In vitro, NVP-CGM097 sulfate demonstrates potent and selective anti-proliferative activity against cancer cells that express wild-type p53. In cell-based assays, it effectively inhibits the interaction between p53 and MDM2 in living cells, as demonstrated by its ability to redistribute wild-type p53 into the cell nucleus with an IC₅₀ of 0.224 μM. This nuclear accumulation of p53 is a hallmark of MDM2 inhibition. The compound shows a significant growth inhibitory effect on HCT116 (p53 WT/WT) colon cancer cells with an IC₅₀ of 454 ± 136 nM. Importantly, it exhibits a 35- to 58-fold selectivity for wild-type p53 cells over p53-null cells, confirming that its anti-proliferative effect is p53-dependent. NVP-CGM097 also shows no significant activity against other protein-protein interactions such as Bcl-2:Bak, Mcl-1:Bak, or XIAP:BIR3, indicating a high degree of target specificity.

In vivo, NVP-CGM097 demonstrates potent antitumor activity in mouse xenograft models. In the MDM2-amplified, p53wt SJSA-1 osteosarcoma model, oral administration of NVP-CGM097 at well-tolerated doses (e.g., 25, 50, 100 mg/kg daily) led to significant tumor growth inhibition and even regression. Efficacy was observed across various dosing schedules (from daily to twice per week). Furthermore, it showed efficacy in other p53wt models, including patient-derived xenografts (PDX) of liposarcoma, AML, and endocrine-resistant breast cancer, particularly in combination with other agents like fulvestrant or CDK4/6 inhibitors .

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In the SJSA-1 human tumor model, NVP-CGM097 sulfate reactivates the p53 dye adjacent to MDM2 and suppresses the response between p53 and MDM2, as indicated by higher p21 mRNA levels, a measure of p53 activity according to pharmacodynamic (PD) indications. It was discovered that when NVP-CGM097 sulfate levels were raised to 30 mg/kg in the tumor-bearing state, p21 mRNA levels rose as well. The biphasic PD response lasts for a maximum of twenty-four hours. Comparable patterns were observed in the mRNA levels of other p53 target genes, including MDM2 and PUMA, which were assessed in concurrent samples. NVP-CGM097 sulfate administration on a daily basis dramatically and dose-dependently enhanced SJSA-1 tumor development. 20 mg/kg is a dose that can help stabilize the condition, and the antibiotic's AUC0–24 is 163 μM.h. The total blood clearance (CL) of NVP-CGM097 following intravenous injection is 5 mL/min/kg for mice, 7 mL/min/kg for rats, 3 mL/min/kg for dogs, and 4 mL/min/kg for monkeys. Dogs have a longer apparent terminal half-life (t1/2) than birds (20 hours), whereas rats and monkeys have a longer t1/2 (6–12 hours). NVP-CGM097 was well absorbed upon prototyping, with Tmax occurring in the range of 1 to 4.5 for all studied species [1].
In vivo, NVP-CGM097 sulfate has demonstrated robust anti-tumor activity in preclinical models. In an SJSA-1 human tumor xenograft model, which is characterized by MDM2 amplification, oral administration of NVP-CGM097 effectively inhibited the interaction between p53 and MDM2 and reactivated the p53 pathway. This was evidenced by a dose-dependent elevation of p21 mRNA levels, a known pharmacodynamic (PD) marker of p53 activity. The increase in p21 mRNA was found to be correlated with the levels of NVP-CGM097 in tumor-bearing rats, and the PD response was biphasic and prolonged for up to 24 hours. Other p53 target genes, such as MDM2 and PUMA, also showed a similar response. Daily oral treatment with NVP-CGM097 significantly inhibited the growth of SJSA-1 tumors in a dose-dependent manner, promoting stable disease at a dose of 20 mg/kg. These results confirm its mechanism of action and its potential as an effective anti-cancer agent.

In vivo, NVP-CGM097 demonstrates potent antitumor activity in mouse xenograft models. In the MDM2-amplified, p53wt SJSA-1 osteosarcoma model, oral administration of NVP-CGM097 at well-tolerated doses (e.g., 25, 50, 100 mg/kg daily) led to significant tumor growth inhibition and even regression. Efficacy was observed across various dosing schedules (from daily to twice per week). Furthermore, it showed efficacy in other p53wt models, including patient-derived xenografts (PDX) of liposarcoma, AML, and endocrine-resistant breast cancer, particularly in combination with other agents like fulvestrant or CDK4/6 inhibitors .
In vitro enzyme assays for NVP-CGM097 sulfate are primarily focused on confirming its binding affinity and selectivity for MDM2. A common method is a fluorescence polarization (FP) assay. In this assay, a fluorescently labeled peptide that mimics the p53 binding domain is incubated with MDM2 protein and varying concentrations of the test compound. The compound competes with the labeled peptide for binding to MDM2. When the labeled peptide is bound to MDM2, its rotational motion is restricted, resulting in a higher polarization signal. When the compound displaces the peptide, the signal decreases. The concentration of compound required to displace 50% of the labeled peptide (IC₅₀) is determined, which for NVP-CGM097 is 1.7 nM. To assess selectivity, similar assays are performed with other proteins, such as MDM4, where the IC₅₀ is 2000 nM. These assays are critical for characterizing the compound's potency and target engagement.

: A standard in vitro cell viability assay for NVP-CGM097 is performed using the CellTiter 96 AQueous One Solution Cell Proliferation Assay (MTS). Cells (e.g., 50,000 GOT1 cells/well) are seeded in 96-well plates and allowed to adhere overnight. They are then treated with a range of concentrations of NVP-CGM097 (e.g., 0.1 nM to 2,500 nM) for a defined period, typically 72 to 96 hours. After treatment, the MTS reagent is added to each well and incubated for 1-4 hours. The absorbance is measured at 492 nm using an ELISA plate reader. The IC50 is calculated by comparing the absorbance of treated wells to vehicle-treated control wells .

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Two pairs of cell lines are used to assess NVP-CGM097 p53-dependent antiproliferative effects: (1) an isogenic pair of HCT116 cell lines either expressing wild-type p53 or knocked-out for the p53 gene and (2) a nonisogenic pair of osteosarcoma cell lines either endogenously expressing wild-type p53 and amplified for MDM2 (SJSA-1 cells) or null for p53 (SAOS-2 cells)[1].
In vitro cell-based assays are used to evaluate the functional activity of NVP-CGM097 sulfate in a cellular context. A primary assay is the measurement of cell proliferation. Cancer cell lines with wild-type p53 (e.g., HCT116, SJSA-1) and p53-null cells (e.g., HCT116 p53-/-) are treated with increasing concentrations of the compound for 72-96 hours. Cell viability is then measured using a standard assay like CellTiter-Glo or MTT. The concentration that inhibits 50% of cell growth (GI₅₀ or IC₅₀) is calculated, demonstrating the compound's potency and selectivity. Another key assay is the measurement of p53 pathway activation. Cells are treated with the compound, and the accumulation of p53 and its downstream targets, such as p21 and MDM2, is measured by western blotting or quantitative PCR (qPCR). The re-localization of p53 from the cytoplasm to the nucleus can also be visualized using immunofluorescence microscopy. These assays provide a comprehensive picture of the compound's mechanism of action in living cells.

A typical in vivo efficacy study involves female athymic nude mice bearing subcutaneous SJSA-1 xenografts. When tumors reach a certain size (e.g., ~200-300 mm³), mice are randomized into treatment groups (n=6-12 per group). NVP-CGM097 is formulated as an oral suspension and administered by oral gavage at various doses (e.g., 25, 50, 100 mg/kg) and schedules (e.g., daily for 14 days, or three times a week). Tumor volumes are measured twice weekly with calipers, and body weight is monitored as a proxy for toxicity. At the end of the study, tumors and plasma are collected for pharmacokinetic (PK) and pharmacodynamic (PD) analysis (e.g., measuring p21 mRNA levels) .
In vivo animal models are essential for evaluating the anti-tumor efficacy and pharmacodynamics of NVP-CGM097 sulfate. A typical study uses a xenograft model in immunocompromised mice, where human tumor cells (e.g., SJSA-1, which has MDM2 amplification) are implanted subcutaneously. Once the tumors reach a certain size, the animals are randomized into treatment groups. The compound is administered orally once daily at various doses. Tumor volume is measured regularly with calipers to monitor growth. In a pivotal study, daily treatment with NVP-CGM097 dose-dependently and significantly inhibited SJSA-1 tumor growth. For pharmacodynamic analysis, animals are euthanized at various time points after dosing, and tumor tissues are collected. The levels of p53 target genes, such as p21, MDM2, and PUMA, are measured by qPCR or western blotting to confirm that the compound is engaging its target and activating the p53 pathway in the tumor. Plasma concentrations of the compound are also measured to establish a pharmacokinetic/pharmacodynamic (PK/PD) relationship.

NVP-CGM097 exhibits favorable pharmacokinetic properties in preclinical species. It is well absorbed after oral administration, with time to maximum concentration (Tmax) ranging from 1 to 4.5 hours across species. The oral bioavailability (%F) is high in mice, rats, and dogs, and moderate in monkeys. The total blood clearance (CL) is low, ranging from 3-7 mL/min/kg, which is 5-10% of hepatic blood flow. The terminal half-life (t1/2) is approximately 6-12 hours in rodents and monkeys, and longer (20 hours) in dogs. A good PK/PD relationship has been established, where plasma drug levels correlate with p53 target gene activation in tumors .
NVP-CGM097 sulfate exhibits favorable pharmacokinetic properties across species. After intravenous (IV) administration, the total blood clearance (CL) is 5 mL/min/kg in mice, 7 mL/min/kg in rats, 3 mL/min/kg in dogs, and 4 mL/min/kg in monkeys. The apparent terminal half-life (t₁/₂) is long in rodents and monkeys (6-12 hours) and is comparatively longer in dogs (20 hours). After oral administration, the compound is well absorbed, with a T_max occurring between 1 and 4.5 hours in all species tested. In the SJSA-1 xenograft model, a dose of 20 mg/kg promoting stable disease was associated with a plasma AUC₀₋₂₄ of 163 μM·h. These PK parameters indicate that NVP-CGM097 is a suitable candidate for oral dosing in preclinical and clinical settings. For research purposes, the compound is supplied as a dry powder and can be dissolved in DMSO for in vitro studies or formulated in appropriate vehicles for in vivo oral gavage.

The primary dose-limiting toxicity identified for NVP-CGM097 in clinical studies (Phase I) is delayed thrombocytopenia (low platelet counts). This is an on-target, mechanism-based toxicity resulting from p53 activation in megakaryocytes, which impairs platelet production. Preclinical studies have shown that NVP-CGM097 is not classified as a hazardous substance in acute toxicity tests, and no significant body weight loss was observed in animal efficacy studies at therapeutic doses. However, like all p53-MDM2 inhibitors, careful monitoring of platelet counts is required. Long-term carcinogenicity and reproductive toxicity studies are standard for such investigational agents .
The toxicological profile of NVP-CGM097 sulfate has been evaluated in preclinical studies. Like other MDM2 inhibitors, its primary on-target toxicity is related to the activation of the p53 pathway in normal tissues. This can lead to effects such as gastrointestinal disturbances, myelosuppression (decreased blood cell counts), and other side effects associated with p53 activation. The compound is a potent inhibitor, so careful dose selection is critical to balance efficacy and toxicity. In the preclinical studies, the compound was generally well-tolerated at efficacious doses. For laboratory safety, standard precautions for handling potent anti-cancer agents should be followed, including the use of personal protective equipment and working in a designated area. The compound is for research use only.

[1]. Discovery of a Dihydroisoquinolinone Derivative (NVP-CGM097): A Highly Potent and Selective MDM2 Inhibitor Undergoing Phase 1 Clinical Trials in p53wt Tumors. J Med Chem. 2015 Aug 27;58(16):6348-58.

NVP-CGM097 sulfate is a highly potent and selective MDM2 inhibitor that functions as an antagonist of the MDM2-p53 protein-protein interaction. It is an orally bioavailable compound that reactivates the p53 pathway in tumors with wild-type p53, leading to cell cycle arrest and apoptosis. Its high selectivity for MDM2 over MDM4 and other protein targets makes it a valuable tool for studying the p53 pathway and a promising anti-cancer agent. The compound was developed by Novartis and has been investigated in clinical trials. The sulfate salt form is used to improve the compound's aqueous solubility and stability. It is a research compound and is not for human therapeutic use. The compound is often referred to as CGM097 sulfate or simply NVP-CGM097.

C38H49CLN4O8S

757.335668325424

756.296

C, 60.27; H, 6.52; Cl, 4.68; N, 7.40; O, 16.90; S, 4.23

1313367-56-4

NVP-CGM097;1313363-54-0;NVP-CGM097 (stereoisomer);2070009-54-8

53262550

Light yellow to yellow solid powder

2

10

9

52

1120

1

CC(C)OC1=C(C=C2CC(=O)N([C@H](C2=C1)C3=CC=C(C=C3)Cl)C4=CC=C(C=C4)N(C)CC5CCC(CC5)N6CCN(C(=O)C6)C)OC.OS(=O)(=O)O

YFLKIFVIZIALIA-GHVGLMRRSA-N

InChI=1S/C38H47ClN4O4.H2O4S/c1-25(2)47-35-22-33-28(20-34(35)46-5)21-36(44)43(38(33)27-8-10-29(39)11-9-27)32-16-14-30(15-17-32)41(4)23-26-6-12-31(13-7-26)42-19-18-40(3)37(45)24-42;1-5(2,3)4/h8-11,14-17,20,22,25-26,31,38H,6-7,12-13,18-19,21,23-24H2,1-5H3;(H2,1,2,3,4)/t26?,31?,38-;/m0./s1

(1S)-1-(4-chlorophenyl)-6-methoxy-2-[4-[methyl-[[4-(4-methyl-3-oxopiperazin-1-yl)cyclohexyl]methyl]amino]phenyl]-7-propan-2-yloxy-1,4-dihydroisoquinolin-3-one;sulfuric acid

NVP-CGM097 sulfate; NVP-CGM-097 (sulfate); NVP-CGM097 sulfate; CGM-097 Sulfate; CGM 097 Sulfate; NVP CGM097 sulfate

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

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

DMSO: ~100 mg/mL (132.0 mM)
H2O: ~100 mg/mL (132.0 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (3.30 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (3.30 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (3.30 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 50 mg/mL (66.02 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.

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