DC-LC3in-D5 works as an autophagy inhibitor by attenuating the lipidation of LC3B. DC-LC3in-D5 binds to LC3B. DC-LC3in-D5 disrupts the LC3B-LBP2 interaction with IC50 of 200 nM. DC-LC3in-D5 may contribute to anti-HCV (hepatitis C virus) or tumor combination studies by inhibiting autophagy.

Physicochemical Properties

Molecular Formula	C19H22CL2N2O3
Molecular Weight	397.295583248138
Exact Mass	396.1
CAS #	2868312-73-4
PubChem CID	162641047
Appearance	White to light yellow solid powder
LogP	2.4
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	5
Heavy Atom Count	26
Complexity	561
Defined Atom Stereocenter Count	0
InChi Key	IYDBJXOPTZDWCZ-UHFFFAOYSA-N
InChi Code	InChI=1S/C19H22Cl2N2O3/c20-16-3-1-2-14(19(16)21)13-10-17(24)15(18(25)11-13)12-22-4-5-23-6-8-26-9-7-23/h1-3,12-13,24H,4-11H2
Chemical Name	5-(2,3-dichlorophenyl)-3-hydroxy-2-(2-morpholin-4-ylethyliminomethyl)cyclohex-2-en-1-one
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 the proteome, DC-LC3in-D5 shows strong selectivity for LC3A/B. In HeLa cells, DC-LC3in-D5 demonstrates significant covalent reactivity and selectivity for LC3A/B [1]. HeLa cells treated with DC-LC3in-D5 (3-30 μM) accumulated p62, inhibited the development of autophagic vacuoles, and disrupted LC3B lipidation [1].
Cell Assay	Western Blot Analysis[1] Cell Types: HeLa cells Tested Concentrations: 3, 10, 30 μM Incubation Duration: 16 hrs (hours) Experimental Results: Pre-treated accumulated Dramatically more p62 than DMSO-treated control samples. Attenuated LC3-I/II lipidation in cells exposed to autophagy inducing conditions.
References	[1]. Inhibition of Autophagy by a Small Molecule through Covalent Modification of the LC3 Protein. Angew Chem Int Ed Engl. 2021 Dec 6;60(50):26105-26114.

Solubility Data

Solubility (In Vitro)

DMSO : 35.71 mg/mL (89.88 mM)

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	2.5170 mL	12.5849 mL	25.1699 mL
	5 mM	0.5034 mL	2.5170 mL	5.0340 mL
	10 mM	0.2517 mL	1.2585 mL	2.5170 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.