Thalidomide-NH-C6-NH-Boc is a ligand (for E3 ligase )-linker conjugate that combines cereblon ligand-based Thalidomide and linker proteins for the synthesis of MI -389 (compound 22). MI-389 is a PROTAC protein degrader based on the multi-target receptor tyrosine kinase inhibitor (TKI) sunitinib .

Physicochemical Properties

Molecular Formula	C24H32N4O6
Molecular Weight	472.534086227417
Exact Mass	472.232
CAS #	2093536-13-9
PubChem CID	134559334
Appearance	Light yellow to yellow solid powder
LogP	2.8
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	11
Heavy Atom Count	34
Complexity	808
Defined Atom Stereocenter Count	0
SMILES	O=C1C2C(=CC=CC=2C(N1C1C(NC(CC1)=O)=O)=O)NCCCCCCNC(=O)OC(C)(C)C
InChi Key	MYIJXYLTQWCASN-UHFFFAOYSA-N
InChi Code	InChI=1S/C24H32N4O6/c1-24(2,3)34-23(33)26-14-7-5-4-6-13-25-16-10-8-9-15-19(16)22(32)28(21(15)31)17-11-12-18(29)27-20(17)30/h8-10,17,25H,4-7,11-14H2,1-3H3,(H,26,33)(H,27,29,30)
Chemical Name	tert-butyl N-[6-[[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]amino]hexyl]carbamate
Synonyms	ThalidomideNHC6NHBoc; Thalidomide NH C6 NH Boc
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	With an EC50 value of 21.3 nM, MI-389 (0-1 μM; 72 hours) inhibits cell growth in a manner similar to sunitinib's cellular efficacy (EC50=17.3 nM)[1]. MI-389 (0-500 nM; 72 hours) rapidly causes dose-dependent GSPT1 destabilization. At 100 nM, it demonstrates total GSPT1 depletion[1].
Cell Assay	Cell Viability Assay[1] Cell Types: Kasumi-1 cells (a c-KIT dependent acute myeloid leukemia ( AML) cell line); GIST-T1 Tested Concentrations: 0-1 μM Incubation Duration: 72 hrs (hours) Experimental Results: Outperform diminished-antiproliferative effect than sunitinib Western Blot Analysis[1] Cell Types: Kasumi-1 cells (a c-KIT dependent acute myeloid leukemia (AML) cell line); GIST-T1 Tested Concentrations: 1 nM, 5 nM, 10 nM, 50 nM, 100 nM, 500 nM Incubation Duration: 4 hrs (hours) Experimental Results: diminished GSPT-1 protein expression.
References	[1]. Translation Termination Factor GSPT1 Is a Phenotypically Relevant Off-Target of Heterobifunctional Phthalimide Degraders. ACS Chem Biol. 2018 Mar 16;13(3):553-560. [2]. Translation Termination Factor GSPT1 Is a Phenotypically Relevant Off-Target of Heterobifunctional Phthalimide Degraders.ACS Chem Biol. 2018 Mar 16;13(3):553-560.

Solubility Data

Solubility (In Vitro)

DMSO : ~100 mg/mL (~211.63 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 2.5 mg/mL (5.29 mM) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.1163 mL	10.5813 mL	21.1627 mL
	5 mM	0.4233 mL	2.1163 mL	4.2325 mL
	10 mM	0.2116 mL	1.0581 mL	2.1163 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.