| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| Other Sizes |
| Targets |
Rb-Raf-1 interaction, apoptosis[1]
RRD-251 directly targets the retinoblastoma tumor suppressor protein (Rb) and the interaction interface between Rb and Raf-1 kinase. By specifically binding to Rb, it disrupts the Rb-Raf-1 complex without affecting other Rb interactions (such as with E2F transcription factors). This selective targeting makes Rb-Raf-1 disruption a potential anticancer strategy. |
|---|---|
| ln Vitro |
In vitro, RRD-251 (10-50 μM; 24 hours) suppresses the proliferation of melanoma[1]. In non-small cell lung cancer cells, RRD-251 (50 μM; 2 hours) suppresses Rb-Raf-1 interaction and Rb phosphorylation [1]. RRD-251 causes cell cycle arrest (20–50 μM; 4 hours) and apoptosis (50 μM; 18 hours)[1]. Apoptosis regulating protein and cell cycle expression are modified by RRD-251[1].
RRD-251 potently inhibits the binding of Raf-1 to Rb with an IC50 value of approximately 0.5 uM as measured by ELISA-based assays. In cellular studies, it disrupts the Rb-Raf-1 interaction at concentrations of 1-5 uM, leading to reduced phosphorylation of Rb by Raf-1 and restoration of Rb's tumor suppressor function. It shows antiproliferative activity against several cancer cell lines including colon and lung cancer cells. |
| ln Vivo |
Melanomas treated with RRD-251 (50 mg/kg; ip; qod; for 14 days) have anti-cancer properties in vivo[1].
In mouse xenograft models, RRD-251 administered intraperitoneally at a dose of 10 mg/kg (every two days for 15 days) significantly suppressed tumor growth without causing obvious toxicity. The compound demonstrated efficacy in inhibiting the growth of HCT116 colon cancer xenografts. No significant weight loss or behavioral changes were observed in treated animals, indicating a favorable therapeutic window. |
| Enzyme Assay |
For in vitro binding assays (ELISA): Coat plates with purified GST-Rb protein. Incubate with His-Raf-1 in the presence of various concentrations of RRD-251 (0.1-20 uM) diluted in PBS containing 0.05% Tween-20 and 1% BSA. After washing, detect bound Raf-1 using anti-His antibody followed by HRP-conjugated secondary antibody. Measure absorbance at 450 nm. Calculate IC50 by nonlinear regression.
|
| Cell Assay |
Cell Proliferation Assay[1]
Cell Types: SK-MEL-28 cells, SK-MEL-5 cells, SK-MEL- 2 cells Tested Concentrations: 10 μM, 20 μM, 50 μM Incubation Duration: 24 hrs (hours) Experimental Results: Inhibited melanoma growth in-vitro. Western Blot Analysis[1] Cell Types: SK-MEL-28 cells, SK-MEL-5 cells, SK -MEL-2 cells Tested Concentrations: 50 μM Incubation Duration: 2 hrs (hours) Experimental Results: demonstrated the depletion of phosphorylated-Rb in RRD-251 treated cells. Apoptosis Analysis[1] Cell Types: SK-MEL-28 cells, SK-MEL-5 cells, SK-MEL-2 cells Tested Concentrations: 50 μM Incubation Duration: 18 hrs (hours) Experimental Results: Induced apoptosis. Cell Cycle Analysis[1] Cell Types: SK-MEL-28 cells, SK-MEL-5 cells Tested Concentrations: 20 μM, 50 μM Incubation Duration: 4 hrs (hours) Experimental Results: Resulted in a dose dependent inhibition of cell cycle progression in SK-MEL-28 and SK-MEL-5 cells, respectively. For cell-based mechanistic studies: Culture cancer cells (e.g., HCT116 or A549) in RPMI-1640 with 10% FBS. Treat cells with RRD-251 at 1, 5, and 10 uM for 24-48 hours. Prepare cell lysates and perform co-immunoprecipitation using anti-Rb antibody followed by western blotting for Raf-1. Alternatively, assess Rb phosphorylation status using phospho-specific antibodies. Measure cell viability by MTT assay after 72-hour treatment. |
| Animal Protocol |
Animal/Disease Models: 8-wk-old female athymic nude mice, with SK-ME-28 xenograft[1]
Doses: 50 mg/kg Route of Administration: intraperitoneal (ip)administration, qod, for 14 days Experimental Results: Inhibits the growth of SK-ME-28 xenograft in nude mice. For in vivo xenograft models: Subcutaneously inject 5×10⁶ HCT116 cells into the flank of athymic nude mice. When tumors reach approximately 100 mm3, randomly assign mice to treatment groups (n=8-10 per group). Administer RRD-251 intraperitoneally at 10 mg/kg in 100 uL of vehicle (10% DMSO, 40% PEG300, 5% Tween-80, 45% saline) every two days for 15 days. Measure tumor volume every three days using calipers. Euthanize mice at study endpoint and collect tumors for immunohistochemistry. |
| ADME/Pharmacokinetics |
No detailed pharmacokinetic data is publicly available for RRD-251. As a small molecule with a molecular weight around 400-500 g/mol and moderate lipophilicity, it is expected to have reasonable oral bioavailability, but intraperitoneal administration has been used in published studies. Metabolic stability and plasma half-life remain to be fully characterized. Preliminary data suggest sufficient systemic exposure for tumor growth inhibition.
|
| Toxicity/Toxicokinetics |
In xenograft studies, RRD-251 at the efficacious dose of 10 mg/kg (every two days) did not cause overt toxicity as judged by body weight maintenance and general animal behavior. Higher doses or more frequent dosing schedules have not been reported. No hematological or biochemical toxicity data is available. Standard safety precautions should be taken when handling this research compound.
|
| References | |
| Additional Infomation |
RRD-251 is a research compound that has not entered clinical trials. It is useful as a chemical probe to study the role of Rb-Raf-1 interaction in cancer biology. The compound is supplied as a lyophilized powder and should be stored at -20degC, protected from light. It is soluble in DMSO (up to 20 mM). This compound is intended for laboratory research only and not for human therapeutic use.
|
| Molecular Formula |
C8H9CL3N2S
|
|---|---|
| Molecular Weight |
271.59
|
| Exact Mass |
269.955
|
| CAS # |
72214-67-6
|
| PubChem CID |
12526767
|
| Appearance |
White to off-white solid powder
|
| Density |
1.47g/cm3
|
| Boiling Point |
345.3ºC at 760 mmHg
|
| Flash Point |
162.6ºC
|
| Index of Refraction |
1.643
|
| LogP |
4.722
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
2
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
14
|
| Complexity |
189
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
COMNQRICZGJVLE-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C8H8Cl2N2S.ClH/c9-6-2-1-5(7(10)3-6)4-13-8(11)12;/h1-3H,4H2,(H3,11,12);1H
|
| Chemical Name |
(2,4-dichlorophenyl)methyl carbamimidothioate;hydrochloride
|
| 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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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) |
DMSO : 125 mg/mL (460.25 mM)
|
|---|---|
| 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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.6820 mL | 18.4101 mL | 36.8202 mL | |
| 5 mM | 0.7364 mL | 3.6820 mL | 7.3640 mL | |
| 10 mM | 0.3682 mL | 1.8410 mL | 3.6820 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
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
