| ln Vitro |
ITRI-90 (0-20 μM, 24 h) reduced the levels of full-length AR (AR-FL) and splice variant AR-V (ΔLBD) proteins in LNCaP, CWR22Rv1, and VCaP cells (DC50 for AR-FL was 2.12, 5.73, and 8.67 nM, respectively, and for AR-V was 4.72 and 0.29 nM, respectively)[1]. ITRI-90 (10 μM, 24 h) showed a long duration of action in LNCaP, CWR22Rv1, and VCaP cells, allowing for a lower dosing frequency[1]. ITRI-90 (10 μM, 6 or 10 h) degraded AR protein levels in CWR22Rv1 cells via a ubiquitin-proteasome-dependent system[1]. ITRI-90 (0.1-1 μM, 24 hours) showed low inhibitory efficiency against the expression of downstream target genes in the AR and AR-V7 signaling pathways. In LNCaP and CWR22Rv1 cells, it mediated AR inhibition by targeting NTD protein degradation or directly blocking ligand binding [1]. ITRI-90 (0.001-100 μM, 24 hours or 7 days) specifically and effectively inhibited cancer cell proliferation and induced apoptosis in LNCaP, CWR22Rv1 and VCaP cells [1]. ITRI-90 (5 or 10 μM, 1 day of treatment) effectively degraded Enzalutamide-induced full-length AR and truncated ARV (ΔLBD) proteins in both short-term and long-term acquired resistance cells, thereby significantly weakening the growth of Enzalutamide-resistant cells in VCaP, C4-2B and C4-2B/EnzR cells [1]. ITRI-90 showed low plasma clearance (6.62 mL/min/kg) and good actual in vivo exposure (AUC/dose = 2502.4) in mouse liver microsomes[1].
|
|---|---|
| ln Vivo |
ITRI-90 (10 mg/kg, intraperitoneal injection, twice daily for 15 days) significantly inhibited tumor growth in the CWR22Rv1 xenograft model and no significant toxicity was observed during tumor regression [1]. ITRI-90 (100 mg/kg, local, twice daily for 21 days) is a vascular-available degrader that indicated full-length AR (AR-FL) and LBD domain-truncated AR variants (AR-Vs) during tumor regression in the CWR22Rv1 xenograft mouse model [1].
|
| Cell Assay |
Western Blot Analysis[1]
Cell Types: LNCaP, CWR22Rv1, VCaP cells Tested Concentrations: 0, 0.01, 0.03, 0.1, 0.3, 1, 3, 10 and 20 μM Incubation Duration: 24 h Experimental Results: Effectively reduced AR-FL and AR-V(ΔLBD) proteins in a dose-dependent manner. Western Blot Analysis[1] Cell Types: LNCaP, CWR22Rv1, VCaP cells Tested Concentrations: 10 μM Incubation Duration: 24 h Experimental Results: Displayed different kinetics of degradation and sustainability after drug washout at different time. Achieved AR and AR-V(ΔLBD) degradation by 24 h treatment. Appeared to sustain low pro tein levels up to 24 h after drug washout. Western Blot Analysis[1] Cell Types: CWR22Rv1 and 293T cells Tested Concentrations: 10 μM Incubation Duration: 6 or 10 h Experimental Results: Degradation of AR and AR-V(ΔLBD) was recovered by MG132 and MLN4924 treatment. Showed AR proteins strongly ubiquitinated. Cell Proliferation Assay[1] Cell Types: LNCaP, CWR22Rv1, VCaP cells Tested Concentrations: 0.001, 0.01, 0.1, 1, 10 and 100 μM Incubation Duration: 7 days Experimental Results: Strongly inhibited cell growth by over 90% with IC50s = 6.587, 4.134, 5.454 μM for LNCaP, CWR22Rv1, VCaP cells, respectively. Activated caspase 3/7 within 24 h. Real Time qPCR[1] Cell Types: LNCaP and CWR22Rv1 cells Tested Concentrations: 0.1 and 1μM Incubation Duration: 24 h Experimental Results: Decreased KLK3, NKX3.1 and TMPRSS2 expression in LNCaP and CWR22Rv1 cells. Reduced CCNA2, CDC20, CDK1, UBE2C, UGT2B17 and EDN2 in CWR22Rv1 cells. Cell Proliferation Assay [1] Cell Types: LNCaP, CWR22Rv1, VCaP cells Tested Concentrations: 0.001, 0.01, 0.1, 1, 10 and 100 μM Incubation Duration: 7 days Experimental Results: Strongly inhibited cell growth by over 90% for LNCaP, CWR22Rv1 and VCaP cells with IC50s = 6.587, 4.134 and 5.454 μM. Apoptosis Analysis[1] Cell Types: LNCaP, CWR22Rv1, VCaP cells Tested Concentrations: 10 μM Incubation Duration: 24 h Experimental Results: Rapidly activated caspase 3/7. Western Blot Analysis[1] Cell Types: VCaP, C4-2B/EnzR cells cells Tested Concentrations: 10 μM Incubation Duration: 24 h Experimental Results: Reversed AR degraders significantly in VCaP cell. Displayed Enzalutamide resistance as well as elevated AR-FL and AR-V7 in Long-term drug exposure of C4-2B Cells resulted in C4-2B/EnzR cells. Efficiently reduced both AR-FL and the truncated AR-Vs. Cell Proliferation Assay[1] Cell Types: VCaP, EnzR cells Tested Concentrations: 5 and 10 μM Incubation Duration: 24 h Experimental Results: Inhibits VCaP proliferation when combined with Enzalutamide. Significantly inhibited EnzR cells proliferation. |
| Animal Protocol |
Animal/Disease Models: CWR22Rv1 cells (5 × 106) induced-male C.B-17 SCID mice (4-6 weeks)[1]
Doses: 10 mg/kg Route of Administration: twice daily for 15 days Experimental Results: Significantly impaired the tumor growth without obvious toxicities[1]. Reached an average tumor growth inhibition (TGI) of 76.64% on day 12[1]. Significantly decreased AR-FL 12 and AR-V protein levels in tumors detected at the final time point[1]. Animal/Disease Models: CWR22Rv1 cells (5 × 106) induced-male C.B-17 SCID mice (4-6 weeks)[1] Doses: 100 mg/kg Route of Administration: p.o., twice daily for 21 days Experimental Results: Observed significant tumor suppression with an average TGI of 71.73 % without obvious toxicity[1]. Reduced protein levels of AR-FL and AR Vs in the tumors[1]. |
| References |
| CAS # |
2798907-16-9
|
|---|---|
| Appearance |
Typically exists as solids at room temperature
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| Solubility (In Vivo) |
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.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] Oral Formulations
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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
