DDO3711, a PP5-recruiting phosphatase recruitment chimeras (PHORCs), is created by joining a PP5 activator and a small molecular apoptosis signal-regulated kinase 1 (ASK1) inhibitor with a chemical linker. DDO3711 specifically inhibits ASK1 while not ASK2 (IC50 > 20 M) (IC50 = 164.1 nM). By enlisting PP5, DDO3711 significantly dephosphorylates p-ASK1T838 and exhibits ASK1-dependent antiproliferative activity. DDO3711 has anti-cancer properties and may be useful for research into abnormally phosphorylated oncoproteins.

Physicochemical Properties

Molecular Formula	C42H41N9O6
Molecular Weight	767.83
Exact Mass	767.317
CAS #	2673364-10-6
Related CAS #	2673364-10-6
PubChem CID	171360934
Appearance	Typically exists as solid at room temperature
LogP	6.1
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	10
Rotatable Bond Count	17
Heavy Atom Count	57
Complexity	1280
Defined Atom Stereocenter Count	0
InChi Key	JLYAXFNOILIKPP-KXQOOQHDSA-N
InChi Code	InChI=1S/C47H55ClF3N5O6S3/c1-46(2)20-18-42(34-8-12-37(48)13-9-34)36(31-46)32-55-22-24-56(25-23-55)39-14-10-35(11-15-39)45(57)53-65(60,61)41-16-17-43(44(30-41)64(58,59)47(49,50)51)52-38(19-21-54-26-28-62-29-27-54)33-63-40-6-4-3-5-7-40/h3-17,30,38,52H,18-29,31-33H2,1-2H3,(H,53,57)/t38-/m1/s1
Chemical Name	(R)-4-(4-((4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((4-((4-morpholino-1-(phenylthio)butan-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide
Synonyms	DDO3711
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)

Biological Activity

Targets	ASK1 (IC50 = 164.1 nM); ASK2 (IC50 >20 μM)
ln Vitro	Attenuated cells treated with DDO3711 (15 μM; 24 hours) show anti-proliferative effects by lowering p-ASK1T838 in a PP5-dependent way. GES-1 and HGC-27 cell growth is not inhibited by DDO3711 [1]. In a concentration-dependent manner, DDO3711 (5-50 μM; 0.5-2 hours) suppresses the expression of CDK4/6 and cyclin D1 [1]. Dephosphorylation. DDO3711 lowers p-JNK and p-p38 levels in a concentration-dependent manner[1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: MKN45 cells Tested Concentrations: 15 μM Incubation Duration: 24 h Experimental Results: demonstrated strong anti-proliferative activity (IC50 =0.5μM). Cell cycle analysis[1] Cell Types: MKN45 cell Tested Concentrations: 5 μM Incubation Duration: 1-24 h Experimental Results: Inhibited the expression of CDK4/6 and cyclin D1 in a concentration-dependent manner. Western Blot Analysis [1] Cell Types: MKN45 Cell Tested Concentrations: 5, 25, 50 μM Incubation Duration: 0.5-2 h Experimental Results: p-ASK1T838 can be dephosphorylated in vitro in a time- and concentration-dependent manner.
Animal Protocol	Animal/Disease Models: 4weeks old BALB/c nude mice MKN45 cells [DDO3711 (20, 40 mg/kg; abdominal bladder; daily; for 21 days) Dramatically inhibited tumor growth in a dose-dependent manner [1]. 1] Doses: 20, 40 mg/kg Route of Administration: IP; daily; continued for 21 days Experimental Results: Significant inhibition of tumor growth in a dose-dependent manner. The levels of p-ASK1T838 were Dramatically diminished.
References	[1]. Protein Phosphatase 5-Recruiting Chimeras for Accelerating Apoptosis-Signal-Regulated Kinase 1 Dephosphorylation with Antiproliferative Activity. J Am Chem Soc. 2022 Dec 22.

Solubility Data

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 (e.g. IP/IV/IM/SC) 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). 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) 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). 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 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.3024 mL	6.5119 mL	13.0237 mL
	5 mM	0.2605 mL	1.3024 mL	2.6047 mL
	10 mM	0.1302 mL	0.6512 mL	1.3024 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.