| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
LXH-3-71 targets PHGDH (3-phosphoglycerate dehydrogenase), which is the first enzyme in the phosphorylated serine synthesis pathway. PHGDH is intricately associated with the regulation of numerous cancer stem cells. As a molecular glue, LXH-3-71 does not inhibit PHGDH's enzymatic activity; instead, it promotes the interaction between PHGDH and the DDB1-CRL E3 ligase complex (a cullin-RING E3 ligase with DDB1 adapter). This leads to polyubiquitination and subsequent proteasomal degradation of PHGDH. By degrading PHGDH, LXH-3-71 reduces the stemness of colorectal cancer cells. The compound covalently binds to Cys281 of PHGDH.
|
|---|---|
| ln Vitro |
In vitro, LXH-3-71 robustly induces degradation of PHGDH in colorectal cancer cells, thereby modulating cancer stem cell properties. The degradation is selective and significant, reducing cancer cell stemness without affecting PHGDH's biosynthetic function. As a molecular glue, it covalently binds to PHGDH at Cys281 and promotes formation of a dynamic chimera with DDB1-CRL E3 ligase, leading to ubiquitination and proteasome-mediated degradation. This mechanism results in decreased proliferation and altered stem-like characteristics in CRC cells. The compound has also been leveraged as a novel ligand for the DDB1-CRL E3 ligase, facilitating the development of new PROTAC molecules targeting EGFR and CDK4 degradation.
|
| ln Vivo |
LXH-3-71 has been demonstrated to be effective in vivo, modulating the stemness of colorectal cancer cells and inhibiting tumor growth in CRC models. It robustly induced degradation of PHGDH, thereby modulating the stemness of colorectal cancer cells (CRCs) in vivo. The compound has been used to develop new PROTAC molecules. Its in vivo efficacy supports further investigation as a potential therapeutic strategy for colorectal cancer by targeting PHGDH degradation rather than its enzymatic inhibition. Specific dosing and route information: details are available in the primary research literature (Acta Pharmaceutica Sinica B, 2024). For research use only.
|
| Enzyme Assay |
Cell-free binding assays confirm that LXH-3-71 covalently binds to PHGDH at Cys281. A typical pull-down assay: recombinant PHGDH protein is incubated with biotinylated LXH-3-71 (0.1-10 uM) in binding buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM DTT) for 1 hour at 4degC. Streptavidin beads are added to capture the protein-ligand complex. After washing, bound proteins are eluted and analyzed by SDS-PAGE and Western blot using anti-PHGDH antibody. For competition, excess unlabeled LXH-3-71 (100 uM) is added. This confirms direct and covalent binding. Mass spectrometry can identify the Cys281 modification. These cell-free assays are key to establishing LXH-3-71 as a molecular glue.
|
| Cell Assay |
For cellular degradation assays, colorectal cancer cells (e.g., HCT-116, HT-29) are seeded in 6-well plates (5 × 10⁵ cells/well) in DMEM with 10% FBS. After 24 hours, cells are treated with LXH-3-71 (0.1-100 uM) for 6-24 hours. For proteasome inhibition, pre-treat with MG132 (10 uM, 1 hour). Cells are lysed in RIPA buffer with protease inhibitors. Protein lysates (30 ug) are resolved by SDS-PAGE and immunoblotted with anti-PHGDH antibody. GAPDH or beta-actin serves as loading control. PHGDH degradation is quantified by densitometry. LC50 values can be calculated. For cancer stemness assays, perform sphere formation assays: cells are plated in ultra-low attachment plates in serum-free DMEM/F12 with growth factors (EGF, bFGF). LXH-3-71 reduces sphere size and number, indicating reduced stemness.
|
| Animal Protocol |
In vivo efficacy is evaluated in a colorectal cancer xenograft model. Female BALB/c nude mice (6-8 weeks, 18-22 g) are subcutaneously injected with 5 × 10⁶ HCT-116 cells in 0.1 mL PBS. When tumors reach 150-200 mm3, mice are randomized into groups (n=8). LXH-3-71 is formulated in 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline and administered via intraperitoneal (IP) injection at doses of 10-50 mg/kg daily for 14-21 days. Tumor volume (V = length × width2 × 0.5) and body weight are measured twice weekly. At study end, tumors are excised, weighed, and processed for Western blot (PHGDH levels) and IHC (Ki-67, cleaved caspase-3). Tumor growth inhibition (TGI) is calculated. This protocol is based on standard in vivo practices for evaluating PHGDH degraders.
|
| ADME/Pharmacokinetics |
Pharmacokinetic properties of LXH-3-71 are not reported in the provided datasheets. As a molecular glue that covalently binds to PHGDH (MW ~456.20), it is likely to have moderate to high oral bioavailability in rodents. The terminal half-life (t1/2) may be extended due to covalent target engagement. For in vivo use, formulate as described above. For storage: powder at -20degC. Stable for 2 years. For in vitro, dissolve in DMSO (50 mg/mL). Do not freeze-thaw repeatedly. Not for human use.
|
| Toxicity/Toxicokinetics |
Preclinical toxicity data for LXH-3-71 are not available. As a research chemical for in vitro and in vivo cancer studies, it should be handled with standard laboratory safety precautions: wear gloves, lab coat, and safety glasses. Avoid inhalation and skin contact. The compound is not intended for human therapeutic use. No acute oral LD50 data reported. Store at -20degC. Dispose of in accordance with local regulations. Potential on-target toxicity may relate to PHGDH degradation in normal tissues, but further studies are needed.
|
| References | |
| Additional Infomation |
LXH-3-71 has the CAS number 2251753-65-6. Molecular formula: C24H28N4O4S, MW 468.57. It is also known as PHGDH degrader, a molecular glue. For research use only. It is described in Acta Pharmaceutica Sinica B, 2024, doi: 10.1016/j.apsb.2024.06.001. The compound covalently binds to Cys281 of PHGDH. It can be used as a ligand for DDB1-CRL E3 ligase to develop new PROTAC molecules targeting EGFR and CDK4. Research applications: colorectal cancer, cancer stem cells, targeted protein degradation. Purity >98%.
|
| Molecular Formula |
C41H59N3O4
|
|---|---|
| Molecular Weight |
657.92
|
| CAS # |
2251753-65-6
|
| 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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5199 mL | 7.5997 mL | 15.1994 mL | |
| 5 mM | 0.3040 mL | 1.5199 mL | 3.0399 mL | |
| 10 mM | 0.1520 mL | 0.7600 mL | 1.5199 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
