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LV-320

Cat No.:V51835 Purity: ≥98%
LV-320 is a potent and noncompetitive ATG4B inhibitor (antagonist) with IC50 of 24.5 μM and a Kd of 16 μM.
LV-320
LV-320 Chemical Structure CAS No.: 2449093-46-1
Product category: Autophagy
This product is for research use only, not for human use. We do not sell to patients.
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Product Description
LV-320 is a potent and noncompetitive ATG4B inhibitor (antagonist) with IC50 of 24.5 μM and a Kd of 16 μM. LV-320 inhibits the enzymatic activity of ATG4B, blocks autophagy, and is stable, non-toxic and active in the body.
Biological Activity I Assay Protocols (From Reference)
Targets
IC50: 24.5 µM (ATG4B); Kd: 16 μM (ATG4B)[1]
ln Vitro
In all four cell lines, endogenous LC3B-II and protein p62 levels increase in a dose-dependent manner upon treatment with LV-320 (0-120 µM; SKBR3, MCF7, JIMT1, and MDA-MB-231 cells)[1]. Treatment with -320 (120 µM; 48 hours; MDA-MB-231 cells) causes an increase in LC3B-II, suggesting that LV-320 inhibits autophagic flux[1].
ln Vivo
Treatment with LV-320 (100-200 mg/kg; oral gavage; three times over two days; GFP-LC3 mice) causes a 104 µM liver level and a 169 µM terminal blood level. Compared to controls, mice treated with LV-320 exhibited a considerably higher accumulation of GFP-LC3 puncta expression. Animals treated with LV-320 also exhibit an increase in LC3B-II protein. At any dosage, the therapy does not significantly harm mice[1].
Cell Assay
Western Blot Analysis[1]
Cell Types: SKBR3, MCF7, JIMT1, and MDA-MB-231 cells
Tested Concentrations: 0 µM, 25 µM, 50 µM, 75 µM, 100 µM, or 120 µM
Incubation Duration:
Experimental Results: Resulted in a dose-dependent increase in endogenous LC3B-II and protein p62 levels in all four cell lines. Cell Autophagy Assay[1]
Cell Types: MDA-MB-231 cells
Tested Concentrations: 120 µM
Incubation Duration: 48 hrs (hours)
Experimental Results: Blocked autophagic flux.
Animal Protocol
Animal/Disease Models: GFP-LC3 mice (females, 9-14 weeks)[1]
Doses: 100 mg/kg or 200 mg/kg
Route of Administration: po (oral gavage); three times over two days (pharmacokinetic/PK study)
Experimental Results: Terminal blood levels were 169 µM and liver levels were 104 µM. LC3B-II protein level was also increased.
References

[1]. A new quinoline-based chemical probe inhibits the autophagy-related cysteine protease ATG4B. Sci Rep. 2018 Aug 3;8(1):11653. doi: 10.1038/s41598-018-29900-x.

These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C29H26CLNO2S2
Molecular Weight
520.11
Exact Mass
519.109
CAS #
2449093-46-1
PubChem CID
134817239
Appearance
Light yellow to yellow solid powder
Density
1.317±0.06 g/cm3(Predicted)
Boiling Point
718.4±60.0 °C(Predicted)
LogP
7.8
Hydrogen Bond Donor Count
1
Hydrogen Bond Acceptor Count
5
Rotatable Bond Count
11
Heavy Atom Count
35
Complexity
663
Defined Atom Stereocenter Count
0
SMILES
C1=CC=C(C=C1)CCSC(C2=CC=C(C=C2)/C=C/C3=C4C=CC(=CC4=NC=C3)Cl)SCCC(=O)O
InChi Key
PHENTWJNXGESKT-RMKNXTFCSA-N
InChi Code
InChI=1S/C29H26ClNO2S2/c30-25-12-13-26-23(14-17-31-27(26)20-25)9-6-22-7-10-24(11-8-22)29(35-19-16-28(32)33)34-18-15-21-4-2-1-3-5-21/h1-14,17,20,29H,15-16,18-19H2,(H,32,33)/b9-6+
Chemical Name
3-[[4-[(E)-2-(7-chloroquinolin-4-yl)ethenyl]phenyl]-(2-phenylethylsulfanyl)methyl]sulfanylpropanoic acid
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)
Solubility Data
Solubility (In Vitro)
DMSO : 135 mg/mL (259.56 mM)
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.25 mg/mL (4.33 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 22.5 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: ≥ 2.25 mg/mL (4.33 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 22.5 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 1.9227 mL 9.6134 mL 19.2267 mL
5 mM 0.3845 mL 1.9227 mL 3.8453 mL
10 mM 0.1923 mL 0.9613 mL 1.9227 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.

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What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
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Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
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In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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Calculation results

Working concentration mg/mL;

Method for preparing DMSO stock solution mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.

Method for preparing in vivo formulation:Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.

(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
             (2) Be sure to add the solvent(s) in order.

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