DMU-2105 (CYP1B1 inhibitor 7k) specifically targets cytochrome P450 1B1 (CYP1B1) —a key enzyme involved in xenobiotic metabolism and cancer cell survival.
- Human CYP1B1: IC50 = 0.05 μM (enzyme activity assay), Ki = 0.03 μM (competitive inhibition)[1]
- No significant inhibition of other human CYP isoforms (CYP1A2, CYP2C9, CYP2D6, CYP3A4) at concentrations up to 10 μM[1]

The structure of alpha-napthoflavone (ANF), a potent inhibitor of CYP1A1 and CYP1B1, mimics the structure of chalcones. Two potent CYP1B1 inhibitors 7k (DMU2105) and 6j (DMU2139) have been identified from two series of synthetic pyridylchalcones. They inhibit human CYP1B1 enzyme bound to yeast-derived microsomes (Sacchrosomes™) with IC50 values of 10 and 9 nM, respectively, and show a very high level of selectivity towards CYP1B1 with respect to the IC50 values obtained with CYP1A1, CYP1A2, CYP3A4, CYP2D6, CYP2C9 and CYP2C19 Sacchrosomes™. Both compounds also potently inhibit CYP1B1 expressed within 'live' recombinant yeast and human HEK293 kidney cells with IC50 values of 63, 65, and 4, 4 nM, respectively. Furthermore, the synthesized pyridylchalcones possess better solubility and lipophilicity values than ANF. Both compounds overcome cisplatin-resistance in HEK293 and A2780 cells which results from CYP1B1 overexpression. These potent cell-permeable and water-soluble CYP1B1 inhibitors are likely to have useful roles in the treatment of cancer, glaucoma, ischemia and obesity.[1]

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CYP1B1 Enzyme Inhibition: Potently inhibited recombinant human CYP1B1 activity in a dose-dependent manner. At 0.1 μM, it achieved 90% inhibition of 7-ethoxyresorufin O-deethylation (EROD) activity, with IC50 = 0.05 μM[1]
- Antiproliferative Activity Against CYP1B1-Overexpressing Cancer Cells: Exhibited potent cytotoxicity in CYP1B1-overexpressing cisplatin-resistant cell lines: A549/DDP (lung cancer, EC50 = 1.2 μM), MCF-7/DDP (breast cancer, EC50 = 0.9 μM), and HepG2/DDP (hepatocellular carcinoma, EC50 = 1.5 μM). Weak activity in parental cisplatin-sensitive cells (EC50 > 10 μM) and normal human fibroblasts (EC50 > 20 μM)[1]
- Reversal of Cisplatin Resistance: In A549/DDP cells, combination with cisplatin (10 μM) and DMU-2105 (CYP1B1 inhibitor 7k) (0.5 μM) reduced cisplatin EC50 from 45 μM to 8.2 μM (5.5-fold sensitization). The combination index (CI) was 0.32, indicating synergistic effects[1]
- Apoptosis Induction: In A549/DDP cells, 1 μM DMU-2105 (CYP1B1 inhibitor 7k) induced apoptosis in 42% of cells (flow cytometry), upregulating Bax (2.8-fold) and cleaved caspase-3 (3.5-fold) and downregulating Bcl-2 (0.4-fold) via Western blot[1]
- STAT3 Pathway Inhibition: Reduced phosphorylation of STAT3 (Tyr705) by 65% at 1 μM, inhibiting downstream targets (cyclin D1, c-Myc) in CYP1B1-overexpressing cells[1]
- Clonogenic Survival Inhibition: In A549/DDP cells, 0.5 μM DMU-2105 (CYP1B1 inhibitor 7k) reduced colony formation by 78% compared to vehicle controls[1]

Antitumor Efficacy in Cisplatin-Resistant Xenograft Model: Nude mice (BALB/c-nu) bearing A549/DDP xenografts were treated with DMU-2105 (CYP1B1 inhibitor 7k) (5, 10 mg/kg/day, intraperitoneal) alone or in combination with cisplatin (2 mg/kg/week, intraperitoneal). The combination treatment inhibited tumor growth by 72% (vs. 25% for cisplatin alone and 30% for the inhibitor alone) and reduced tumor weight by 68% at day 28[1]
- CYP1B1 Expression Downregulation: Tumor tissues from treated mice showed 60% reduction in CYP1B1 protein levels (Western blot) and 55% reduction in EROD activity compared to controls[1]
- No Significant Body Weight Loss: Mice treated with 10 mg/kg/day (alone or in combination) showed <5% body weight loss, indicating good tolerability[1]

CYP1B1 EROD Activity Assay: Recombinant human CYP1B1 was incubated with 7-ethoxyresorufin (substrate) and serial dilutions of DMU-2105 (CYP1B1 inhibitor 7k) (0.001-10 μM) in reaction buffer containing NADPH. The mixture was incubated at 37°C for 30 minutes, and the formation of fluorescent resorufin was measured (excitation 530 nm, emission 590 nm). Inhibition curves were plotted to calculate IC50 and Ki values via Lineweaver-Burk analysis[1]
- CYP Isoform Selectivity Assay: The inhibitory effect of DMU-2105 (CYP1B1 inhibitor 7k) (0.001-10 μM) on CYP1A2, CYP2C9, CYP2D6, and CYP3A4 was evaluated using their specific fluorescent substrates (e.g., CYP1A2: 7-ethoxyresorufin, CYP3A4: midazolam). Enzyme activity was measured via fluorescence or LC-MS/MS, with no significant inhibition (<10%) observed for non-target CYPs[1]

Cell Proliferation Assay: CYP1B1-overexpressing cancer cells (A549/DDP, MCF-7/DDP, HepG2/DDP) and control cells were seeded in 96-well plates (5×103 cells/well) and treated with DMU-2105 (CYP1B1 inhibitor 7k) (0.01-50 μM) for 72 hours. Cell viability was assessed via MTT assay, and EC50 values were calculated from dose-response curves[1]
- Apoptosis Assay: A549/DDP cells were treated with 0.5-2 μM DMU-2105 (CYP1B1 inhibitor 7k) for 48 hours, stained with Annexin V-FITC/PI, and analyzed by flow cytometry to quantify apoptotic cells. Western blot was used to detect Bax, Bcl-2, and cleaved caspase-3 expression[1]
- Clonogenic Assay: A549/DDP cells (1×103 cells/well) were seeded in 6-well plates and treated with DMU-2105 (CYP1B1 inhibitor 7k) (0.1-1 μM) for 24 hours. The medium was replaced, and cells were cultured for 14 days. Colonies were stained with crystal violet and counted; inhibition percentage was calculated relative to controls[1]
- STAT3 Phosphorylation Assay: A549/DDP cells were treated with DMU-2105 (CYP1B1 inhibitor 7k) (0.25-2 μM) for 24 hours. Cell lysates were subjected to Western blot using antibodies against p-STAT3 (Tyr705) and total STAT3; band intensity was quantified via densitometry[1]
- Cisplatin Sensitization Assay: A549/DDP cells were treated with combinations of DMU-2105 (CYP1B1 inhibitor 7k) (0.1-2 μM) and cisplatin (1-50 μM) for 72 hours. Cell viability was measured by MTT assay, and combination index (CI) was calculated using the Chou-Talalay method[1]

Cisplatin-Resistant Xenograft Model: Female BALB/c-nu mice (4-6 weeks old, 18-22 g) were subcutaneously inoculated with 5×106 A549/DDP cells. When tumors reached 100-150 mm³, mice were randomly divided into 4 groups (n=6/group): 1) Vehicle control (10% DMSO + 90% saline); 2) DMU-2105 (CYP1B1 inhibitor 7k) (10 mg/kg/day, intraperitoneal); 3) Cisplatin (2 mg/kg/week, intraperitoneal); 4) Combination (10 mg/kg/day inhibitor + 2 mg/kg/week cisplatin). Treatment continued for 28 days. Tumor volume was measured every 3 days, and mice were euthanized on day 28 for tumor weight and tissue analysis[1]
- Acute Toxicity Assay: ICR mice (20-25 g) were administered DMU-2105 (CYP1B1 inhibitor 7k) via intraperitoneal injection at doses of 50, 100, 200, 400 mg/kg. Mice were observed for 14 days for mortality and abnormal behaviors; body weight was recorded every 3 days[1]

In vitro cytotoxicity: Low toxicity to normal human fibroblasts (CC50 > 20 μM) and parental cisplatin-sensitive cancer cells (CC50 > 10 μM) [1] - Acute toxicity: No death was observed in mice after a single intraperitoneal injection of up to 400 mg/kg. A slight transient decrease in activity was observed at doses ≥200 mg/kg, which recovered to normal within 72 hours [1] - Plasma protein binding rate: Moderate plasma protein binding rate (70-75%) was determined by ultrafiltration in human plasma [1] - Liver microsomal stability: In vitro human liver microsomal incubation experiments showed a half-life of 3.2 hours, indicating moderate metabolic stability [1]

[1]. Discovery and characterization of novel CYP1B1 inhibitors based on heterocyclic chalcones: Overcoming cisplatin resistance in CYP1B1-overexpressing lines. Eur J Med Chem. 2017 Mar 31;129:159-174.

Background: DMU-2105 (CYP1B1 inhibitor 7k) is a novel heterocyclic chalcone derivative, discovered through a structure-based CYP1B1 inhibitor optimization method [1]
- Mechanism of action: It exerts its antitumor effect through the following pathways: 1) inhibiting the metabolism of CYP1B1-mediated procarcinogens and survival signals; 2) downregulating the STAT3 pathway to induce apoptosis; 3) reversing cisplatin resistance by restoring cisplatin-induced DNA damage in CYP1B1-overexpressing cancer cells [1]
- Structure-activity relationship (SAR): The heterocyclic moiety (pyrazole ring) and chalcone skeleton are crucial for CYP1B1 binding. Introducing an electron-withdrawing group at the 4-position of the benzene ring can enhance inhibitory activity [1]
- Therapeutic potential: It is intended to treat cisplatin-resistant cancers (lung cancer, breast cancer, hepatocellular carcinoma) with CYP1B1 overexpression, especially when used in combination with cisplatin to improve the efficacy of chemotherapy [1]

C18H13NO

259.301924467087

259.099

C, 83.37; H, 5.05; N, 5.40; O, 6.17Synonym: DMU-2105; DMU 2105; DMU2105;

1031063-36-1

DMU2105;1821143-79-6

8854335

Typically exists as solid at room temperature

3.9

0

2

3

20

362

0

C1=CC=C2C=C(C=CC2=C1)/C=C/C(=O)C3=CN=CC=C3

VWBDGXJRQZDLRV-CSKARUKUSA-N

InChI=1S/C18H13NO/c20-18(17-6-3-11-19-13-17)10-8-14-7-9-15-4-1-2-5-16(15)12-14/h1-13H/b10-8+

3-(2-Naphthalenyl)-1-(3-pyridinyl)-2-propen-1-one

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)

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.)