TCID 30675-13-9

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Molecular Weight: 283.92 TCID is a DUB Inhibitor for Ubiquitin C-terminal hydrolase L3 with IC50 of 0.6 μM, 125-fold selective to L1.

TCID is selective for UCH-L3 over UCH-L1 by over 100-fold. NU6027 (10 μM) does not promote YFP-GLT-1 accumulation in intracellular
vesicles in transfected MDCK cells, while LDN-57444 (10 μM) promotes
YFP-GLT-1 accumulation. NU6027 (10 μM) does not produce long-term
lysosomal degradation of GLT-1, while LDN-57444 (10 μM) has such effect. TCID (10 μM) diminishes GlyT2 ubiquitination in
brainstem and spinal cord primary neurons, which is more pronounced when
UCHL1 is inhibited and when cells are exposed to these Inhibitors for
longer periods.

Protocol(Only for Reference)

Kinase Assay: [1]

Enzyme Inhibition Studies

Various concentrations (100 μM to 50 nM) of TCID dissolved in DMSO are each added to 2 mL of 1 nM UCH-L1 or 50 pM UCH-L3 in UCH reaction buffer and incubated at 25℃ for 30 min. Ubiquitin C-terminal hydrolysis activity is initiated by adding 2 μL of 50 μM ubiquitin-AMC. The reactions are monitored at 25℃ using a Hitachi F4500 fluorescence spectrophotometer. The AMC fluorophore is excited at 380 nm and the rates of release of free AMC are measured by monitoring the increase in fluorescence emission at 460 nm. The initial velocity Vo is measured for each reaction and plotted against TCID concentrations in order to determine the IC50 values using GraFit V.5 software.

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

Species	Baboon	Dog	Monkey	Rabbit	Guinea pig	Rat	Hamster	Mouse
Weight (kg)	12	10	3	1.8	0.4	0.15	0.08	0.02
Body Surface Area (m2)	0.6	0.5	0.24	0.15	0.05	0.025	0.02	0.007
Km factor	20	20	12	12	8	6	5	3

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×	mouse Km(3)	= 11.2 mg/kg
	rat Km(6)

Chemical Information

Molecular Weight (MW)	283.92
Formula	C9H2Cl4O2
CAS No.	30675-13-9

Storage	3 years -20℃Powder
	6 months-80℃in solvent (DMSO, water, etc.)
Synonyms

Solubility (25°C) *	In vitro	DMSO	23 mg/mL (81 mM)
		Water	<1 mg/mL (
		Ethanol	<1 mg/mL (

Chemical Name	1H-Indene-1,3(2H)-dione, 4,5,6,7-tetrachloro-

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Product Categories : Ubiquitin > DUB Inhibitor