IMM-01 is a formin agonist that can inhibit DID-DAD (clear inhibitory domain and clear autoregulatory domain) binding with IC50 of 140 nM. IMM-01 acts by disrupting the autoinhibitory bond between the DID and DAD domains, thereby activating formin. IMM-01 has anti-cancer effects.

Physicochemical Properties

Molecular Formula	C12H17N3O2S
Molecular Weight	267.347281217575
Exact Mass	267.104
CAS #	218795-74-5
PubChem CID	135445526
Appearance	Light yellow to light brown solid powder
LogP	1.9
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	3
Heavy Atom Count	18
Complexity	315
Defined Atom Stereocenter Count	0
SMILES	N(C(NC(C)(C)C)=S)N=CC1=CC=C(O)C=C1O
InChi Key	LTFUAYRGVLQXKC-NTUHNPAUSA-N
InChi Code	InChI=1S/C12H17N3O2S/c1-12(2,3)14-11(18)15-13-7-8-4-5-9(16)6-10(8)17/h4-7,16-17H,1-3H3,(H2,14,15,18)/b13-7+
Chemical Name	1-tert-butyl-3-[(E)-(2,4-dihydroxyphenyl)methylideneamino]thiourea
Synonyms	IMM01; IMM 01; IMM-01
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

ln Vitro	In SW480 circles, IMM-01 (100 μM; 1 hour) detects microtubule stabilization [1]. The LacZ expression in NIH 3T3-SRE-LacZ cells is greatly induced by IMM-01 (10 μM) [1]. In NIH 3T3 cells and SW480 cells, IMM-01 (1-100 μM; 48 hours) activates caspase-3 channels during induction [1].
ln Vivo	IMM-01 (5-25 mg/kg; intravenously; twice weekly; for 3 weeks) suppresses tumor growth in a hot water tumor xenograft model [1].
Cell Assay	Apoptosis analysis [1] Cell Types: NIH 3T3 cells and SW480 cells Tested Concentrations: 1 μM, 10 μM or 100 μM Incubation Duration: 48 hrs (hours) Experimental Results: Induced caspase-3 cleavage.
Animal Protocol	Animal/Disease Models: Athymic nude female mice (6 to 8 weeks) were implanted with SW480 cells [1]. Doses: 5 mg/kg, 25 mg/kg. Route of Administration: intravenous (iv) (iv)injection; twice a week; for 3 weeks. Experimental Results: Slows tumor growth in a dose-dependent manner when injected intravenously (iv) (iv)(iv) through the tail vein.
References	[1]. Small-molecule intramimics of formin autoinhibition: a new strategy to target the cytoskeletal remodeling machinery in cancer cells. Cancer Res. 2013 Nov 15;73(22):6793-803.

Solubility Data

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	3.7404 mL	18.7021 mL	37.4042 mL
	5 mM	0.7481 mL	3.7404 mL	7.4808 mL
	10 mM	0.3740 mL	1.8702 mL	3.7404 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.