HyT36 is a low molecular weight hydrophobic tag that promotes the degradation of fusion proteins and pseudokinase Her3.

Physicochemical Properties

Molecular Formula	C25H44CLNO3
Molecular Weight	442.0748
Exact Mass	441.3
CAS #	1323151-45-6
PubChem CID	52953469
Appearance	Light yellow to brown ointment
LogP	6.1
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	16
Heavy Atom Count	30
Complexity	471
Defined Atom Stereocenter Count	1
SMILES	C[C@H](CCC12CC3CC(C1)CC(C3)C2)C(=O)NCCOCCOCCCCCCCl
InChi Key	FRQCQVUXVOJKTG-AXYUOHRYSA-N
InChi Code	InChI=1S/C25H44ClNO3/c1-20(6-7-25-17-21-14-22(18-25)16-23(15-21)19-25)24(28)27-9-11-30-13-12-29-10-5-3-2-4-8-26/h20-23H,2-19H2,1H3,(H,27,28)/t20-,21?,22?,23?,25?/m1/s1
Chemical Name	(2R)-4-(1-adamantyl)-N-[2-[2-(6-chlorohexoxy)ethoxy]ethyl]-2-methylbutanamide
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 HEK293 cells expressing GFP-HaloTag2/Fz4-HaloTag2/GFP-HaloTag7 fusion protein, HyT36 (10 µM; 24 h) decreases the expression of GFP-HaloTag2/Fz4-HaloTag2 [1]. In HEK293 Flp-In cells expressing HA-EGFP-HaloTag2, HyT36 (10 µM; 2 h or 48 h) decreased EGFP expression and increased Hsp70 binding to the HA-EGFP-HaloTag2 fusion protein [3]. In HEK293 Flp-In cells expressing HA-EGFP-HaloTag2, HyT36 (50 nM; 24 h) lowers the amount of EGFP-HaloTag2 [3]. In NIH-3T3 cells expressing HA−HaloTag2−HRasG12V, HyT36 (1 µM; 4 d) decreases the expression of HA−HaloTag2−HRasG12V [
Cell Assay	Western Blot Analysis[1][3] Cell Types: HEK293 cells expressing GFP-HaloTag2/Fz4-HaloTag2/HA-EGFP-HaloTag2 or NIH-3T3 cells expressing HA−HaloTag2−HRasG12V. Tested Concentrations: 1 µM or 10 µM. Incubation Duration: 2 h, 24 h or 4 days. Experimental Results: diminished the expression of fusion proteins and increased the binding of Hsp70 and HA-EGFP-HaloTag2 fusion protein.
References	[1]. Identification of hydrophobic tags for the degradation of stabilized proteins. Chembiochem. 2012 Mar 5;13(4):538-41. [2]. Poloxin-2HT+: changing the hydrophobic tag of Poloxin-2HT increases Plk1 degradation and apoptosis induction in tumor cells. Org Biomol Chem. 2019 Mar 20;17(12):3113-3117. [3]. A bidirectional system for the dynamic small molecule control of intracellular fusion proteins. ACS Chem Biol. 2013 Oct 18;8(10):2293-2300.

Solubility Data

Solubility (In Vitro)

DMSO: 100 mg/mL (226.21 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.66 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.66 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. 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 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (5.66 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. 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 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.2621 mL	11.3104 mL	22.6209 mL
	5 mM	0.4524 mL	2.2621 mL	4.5242 mL
	10 mM	0.2262 mL	1.1310 mL	2.2621 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.