| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| Other Sizes |
| Targets |
Ki: 71 nM (MIF)[1]
MIF (Macrophage Migration Inhibitory Factor). |
|---|---|
| ln Vitro |
At 2 μM and 0.2 μM, MD13 degrades MIF by 91±5% and 71±7%, respectively. By binding to E3 ligase cereblon, MD13 causes MIF degradation[1]. A549 cancer cells' ability to proliferate is inhibited by MD13 (0–20 μM; 72 hours)[1]. In A549 cells, MD13 (1–5 μM; 48 h) stops the cell cycle at the G2/M phase[1]. ERK signalling is inhibited by MD13 (2 μM; 6-48 h)[1].
MD13 is a MIF-directed PROTAC degrader with a Ki of 71 nM and DC50 <100 nM. It induces MIF degradation with a Dmax (maximum degradation) of 92% after 6 hours of treatment. MD13 reduces cellular MIF protein levels, arrests cells at the G2/M phase, and inhibits the proliferation of A549 lung carcinoma cells. It also inhibits the growth of A549 cell 3D spheroids, demonstrating anti-cancer activity. |
| ln Vivo |
No in vivo activity data for MD13 is publicly available. PROTAC degraders typically show improved in vivo efficacy compared to traditional inhibitors due to sustained target degradation. MIF degradation in vivo could potentially reduce tumor growth, inflammation, and metastasis in murine models, but specific data for MD13 is not reported.
|
| Enzyme Assay |
Not available. A generic MIF binding affinity assay for PROTACs is performed by surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC). For SPR: Recombinant human MIF is immobilized on a sensor chip. MD13 (0.1 nM-10 microM) is injected in running buffer (PBS-T, pH 7.4, 0.01% DMSO). Association and dissociation rates are measured, and Ki (71 nM) is calculated (KD = koff/kon). For degradation assays, HEK293T cells expressing cereblon are treated with MD13 (0-20 microM, 1-24 h), and MIF protein levels are measured by Western blot using anti-MIF antibody. DC50 and Dmax (92% at 6 h) are determined by densitometry.
|
| Cell Assay |
Western Blot Analysis[1]
Cell Types: A549 cells Tested Concentrations: 0-20 μM Incubation Duration: 12 h Experimental Results: Caused depletion of MIF protein. Western Blot Analysis[1] Cell Types: A549 cells Tested Concentrations: 2 μM Incubation Duration: 6, 24 or 48 h Experimental Results: Inhibited ERK phosphorylation. Cell Proliferation Assay[1] Cell Types: A549 cells Tested Concentrations: 0-20 μM Incubation Duration: 72 h Experimental Results: Inhibited the growth of A549 cells in a dose-dependent manner. The inhibitory effect reached about 50% inhibition of cell proliferation at a concentration of 20 µM. Cell Cycle Analysis[1] Cell Types: A549 cells Tested Concentrations: 1, 2, or 5 µM Incubation Duration: 48 h Experimental Results: Dose-dependently induced cell cycle arrest at the G2 /M phase. A549 human lung carcinoma cells are seeded in 6-well plates (2×10⁵/well) in RPMI-1640 with 10% FBS and incubated for 24 h. Cells are treated with MD13 (0-20 microM) for 6-24 h. Cell lysates are collected and analyzed by Western blot with anti-MIF, anti-cereblon, and anti-beta-actin (loading control) antibodies. For proliferation assays, A549 cells are seeded in 96-well plates (5×103/well) and treated with MD13 (0-20 microM) for 48-72 h. Cell viability is measured by MTT or CellTiter-Glo. For cell cycle analysis, A549 cells are treated with MD13 (0-10 microM) for 24-48 h, fixed with 70% ethanol, stained with propidium iodide, and analyzed by flow cytometry for cell cycle distribution (G1, S, G2/M phases). For 3D spheroid assays, A549 cells are seeded in ultra-low attachment 96-well plates (1×103/well) and cultured for 7 days to form spheroids, then treated with MD13 for 72 h. Spheroid size is measured by light microscopy and image analysis. |
| Animal Protocol |
No animal protocol is published. Generic protocol for PROTAC degraders in vivo: female BALB/c nude mice (6-8 weeks) are injected subcutaneously with A549 cells (5×10⁶ in 100 microL PBS). When tumors reach ~100 mm3, mice are randomized (n=8-10/group). MD13 is formulated in 10% DMSO/40% PEG300/5% Tween-80/45% saline or 0.5% methylcellulose and administered intraperitoneally or orally at 10-50 mg/kg daily for 2-3 weeks. Tumor volumes are measured biweekly. At termination, tumors are harvested and analyzed for MIF protein levels by Western blot and immunohistochemistry (to confirm target degradation in vivo). Plasma and tissue samples are collected for PK/PD analysis.
|
| ADME/Pharmacokinetics |
No PK data for MD13 is publicly available. Generic PK for PROTAC molecules (MW ~650-1000 Da): oral bioavailability is often low (<20%) due to high molecular weight, polar surface area, and efflux transporter liability (P-gp, BCRP). Half-life is typically short (1-3 h) in mice. Intraperitoneal or intravenous administration may be preferred for in vivo studies. Degradation activity may persist beyond PK half-life due to sustained MIF protein turnover delay. Protein binding is typically high (>90%).
|
| Toxicity/Toxicokinetics |
No toxicity data reported for MD13. Generic toxicity for PROTACs: ICR mice (5/sex/group) receive a single intraperitoneal dose of MD13 at 10, 30, 100 mg/kg. Animals are observed for 14 days for mortality, clinical signs, body weight, and food consumption. At termination, blood is collected for hematology (CBC, differential) and serum chemistry (ALT, AST, BUN, creatinine, total protein). Gross necropsy and histopathology of major organs (liver, kidney, spleen, heart, lung, GI tract, brain) are performed. Potential toxicities associated with MIF degradation (e.g., immune modulation) and cereblon engagement (e.g., thalidomide-like developmental toxicity) should be evaluated.
|
| References | |
| Additional Infomation |
MD13 is the first reported MIF-directed PROTAC degrader, representing a novel therapeutic strategy that moves beyond traditional tautomerase inhibition to disrupt key inflammation- and cancer-related protein-protein interactions involving MIF. Developed by Dekker and colleagues, MD13 is a research tool for MIF biology and for validating MIF degradation as a therapeutic approach. MD13 is not FDA-approved and has not entered clinical trials. It is protected under license from Arvinas (PROTAC® is a registered trademark of Arvinas Operations, Inc.). MD13 is available for laboratory research use only.
|
| Molecular Formula |
C35H35N5O8
|
|---|---|
| Molecular Weight |
653.68
|
| Exact Mass |
653.248
|
| CAS # |
2758431-97-7
|
| PubChem CID |
163342432
|
| Appearance |
Typically exists as solid at room temperature
|
| LogP |
4.2
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
12
|
| Heavy Atom Count |
48
|
| Complexity |
1240
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1CC(=O)NC(=O)C1N2C(=O)C3=C(C2=O)C(=CC=C3)NCCCCCCCC(=O)NC4=CC=C(C=C4)N5CC6=C(C=C(C=C6)O)OC5=O
|
| InChi Key |
HAHDZDUOFHMMEA-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C35H35N5O8/c41-24-15-10-21-20-39(35(47)48-28(21)19-24)23-13-11-22(12-14-23)37-29(42)9-4-2-1-3-5-18-36-26-8-6-7-25-31(26)34(46)40(33(25)45)27-16-17-30(43)38-32(27)44/h6-8,10-15,19,27,36,41H,1-5,9,16-18,20H2,(H,37,42)(H,38,43,44)
|
| Chemical Name |
8-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]amino]-N-[4-(7-hydroxy-2-oxo-4H-1,3-benzoxazin-3-yl)phenyl]octanamide
|
| 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 | 1.5298 mL | 7.6490 mL | 15.2980 mL | |
| 5 mM | 0.3060 mL | 1.5298 mL | 3.0596 mL | |
| 10 mM | 0.1530 mL | 0.7649 mL | 1.5298 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
