Litronesib

Alias: LY2523355; KF 89617; LY-2523355; KF89617; LY 2523355; KF-89617; Eg5 KinesinRelated Motor Protein Inhibitor

Cat No.:V4905 Purity: ≥98%

COA Product Data Instructions for use SDS

Litronesib (formerly also known as LY2523355; LY-2523355; KF-8961) is a novel, potent and selective inhibitor of the kinesin-related motor protein Eg5 with potential anticancer activity.

Litronesib Chemical Structure CAS No.: 910634-41-2
Product category: Kinesin

This product is for research use only, not for human use. We do not sell to patients.

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Other Forms of Litronesib:

Litronesib Racemate

Official Supplier of:

Top Publications Citing lnvivochem Products

Nature 2021, 597(7874):119-125.

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Nature 597, 119–125 (2021)

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Science Trans Med 2023, 15: eadd7872.

Cancer Cell 2021,39(4):529-547.e7.

Cancer Discov 2022,12(4):1106-1127.

Immunity 2023,56(3):620-634.e11.

J Am Chem Soc 2022,144:21831-21836.

Cell 2020,183:1202-1218.e25.

Science Adv 2022:8,eabm9427.

Nature 2021,597(7874):119-125.

Cell 2020,183:1202-1218.e25.

PNAS Nexus 2022,1(3):pgac063.

ACS Nano 2023,17(10):9110-9125.

Biomaterial 2023 Sep16:302:122333.

Purity & Quality Control Documentation

Current batch：

Purity: ≥98%

COA

MSDS

Instructions

Product Description
Bioactivity & Protocols
Physicochemical Properties
Solubility Data
Calculators
Clinical Trial Information
Biological Data

Product Description

Litronesib (formerly also known as LY2523355; LY-2523355; KF-8961) is a novel, potent and selective inhibitor of the kinesin-related motor protein Eg5 with potential anticancer activity. Litronesib specifically inhibits the activity of Eg5, which in actively dividing tumor cells may cause disruption of mitosis, apoptosis, and ultimately cell death. Additionally referred to as KIF11 or kinesin spindle protein-5, the ATP-dependent Eg5 kinesin-related motor protein is a plus-end directed kinesin motor protein that is crucial for the regulation of spindle dynamics, including assembly and maintenance, during mitosis. Look for open or closed clinical trials that are utilizing this agent. (NCI Lexicon).

Biological Activity I Assay Protocols (From Reference)

Targets

Eg5 (IC50 = 14 μM)

ln Vitro

Litronesib (LY-2523355; KF-89617) is a selective Eg5 inhibitor. Litronesib (25 nM) induces cancer cells to death during mitotic arrest, and this needs sustained activation of spindle-assembly checkpoint (SAC)[1].

ln Vivo

Litronesib (LY2523355; KF89617; 1.1, 3.3, 10, and 30 mg/kg, i.v.) shows antitumor activity in a dose-dependently, and causes a dramatic increase in cancer cells immuno-positive for histone H3 phosphorylation in Colo205 xenograft tumors[1].

Cell Assay

In 96-well plates coated with poly-d-lysine, cancer cells are plated and incubated for an entire night. Following that, cells are given different lengths of time to respond to the indicated concentrations of lidocaine. After that, the cells are permeabilized for 10 minutes with cold methanol, fixed for 45 minutes with 3.7% formaldehyde in PBS or 30 minutes with 1× Prefer fixative solution at room temperature, and then fixed for another 10 minutes with 0.2% Triton X-100 in PBS. PBS is used to wash the cells three times. Following an hour of incubation with 100 μg/mL DNase-free RNase and 10 μg/mL propidium iodide, the cells are scanned to determine the mitotic index (MI), which is calculated as the percentage of cells with condensed DNA. Cells are cultivated with anti-phospho-histone H3 antibody or anti-phospho-histone H2AX at 1:1,000 dilution with 5% bovine serum albumin (BSA) in PBS for an overnight period at 4°C in MI based on histone H3 phosphorylation or apoptosis analysis inspection. Alexa 488 secondary antibody (1:1,000 in PBS-2% BSA) is incubated on cells for 60 minutes at room temperature after they have been washed three times with 0.2% Triton-X 100 in PBS. The cells are staining for 15 minutes in PBS containing 10 μg/mL propidium iodide and 100 μg/mL RNaseA after being rinsed three times with purified water. In Acumen Explorer eX3 microplate cytometer, the stained cells are scanned. Phospho-histone H3-Ser10 or phospho-histone H2AX positivity as a percentage of cells is used to express the results[1].

Animal Protocol

In nude mice, primary human tumor xenograft models are created and kept alive. Tumor volume is measured serially using caliper measurements to determine the antitumor efficacy in subcutaneous xenograft tumor-bearing mice (10 mice per treatment group) derived from either established cancer cell lines or fragments of human tumor explants. Female BDF1 mice weighing 20–23 g are used to create the p388 syngeneic tumor model for high-content imaging analysis. The mice are house-acclimated for a week prior to use in experiments. The STR assay is used to validate the p388 murine lymphocytic leukemia cells, which are then cultured in RPMI1640 medium with 10% FBS. Three rounds of serum-free medium washing precede the intraperitoneal injection of 1.25 million cells into mice for inoculation. Litronesib is administered to mice on the fifth post-implantation day, either as an intravenous bolus or as an intravenous infusion, at the prescribed doses and times. Following the death of the mice, the ascitic (intraperitoneal) fluid containing the p388 tumor cells is extracted and subjected to acumen, flow cytometry, and TUNEL assays for apoptosis, G2-M, and phospho-histone H3. Blood samples are obtained via cardiac puncture for pharmacokinetic analysis. EDTA is used to create plasma, and the amount of lidotrexeb in the plasma is measured[1].

References

[1]. A Novel Eg5 Inhibitor (LY2523355) Causes Mitotic Arrest and Apoptosis in Cancer Cells and Shows Potent Antitumor Activity in Xenograft Tumor Models. Mol Cancer Ther. 2015 Nov;14(11):2463-72.

Additional Infomation

Litronesib has been used in trials studying the treatment of Solid Tumors, Ovarian Cancer, Gastric Cancer, Prostate Cancer, and Acute Leukaemia, among others.
Litronesib is an inhibitor of the kinesin-related motor protein Eg5 with potential antineoplastic activity. Litronesib selectively inhibits the activity of Eg5, which may result in mitotic disruption, apoptosis and consequently cell death in tumor cells that are actively dividing. The ATP-dependent Eg5 kinesin-related motor protein (also known as KIF11 or kinesin spindle protein-5) is a plus-end directed kinesin motor protein that plays an essential role during mitosis, particularly in the regulation of spindle dynamics, including assembly and maintenance.

These protocols are for reference only. InvivoChem does not independently validate these methods.

Physicochemical Properties

Molecular Formula	C23H37N5O4S2
Molecular Weight	511.7010
Exact Mass	511.229
Elemental Analysis	C, 53.99; H, 7.29; N, 13.69; O, 12.51; S, 12.53
CAS #	910634-41-2
Related CAS #	Litronesib Racemate;546111-97-1
PubChem CID	25167017
Appearance	White to off-white solid powder
Density	1.23 g/cm3
LogP	4.499
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	11
Heavy Atom Count	34
Complexity	865
Defined Atom Stereocenter Count	1
SMILES	C([C@]1(SC(NC(=O)C(C)(C)C)=NN1C(=O)C(C)(C)C)C1C=CC=CC=1)NS(=O)(=O)CCNCC
InChi Key	YVAFBXLHPINSIK-QHCPKHFHSA-N
InChi Code	InChI=1S/C23H37N5O4S2/c1-8-24-14-15-34(31,32)25-16-23(17-12-10-9-11-13-17)28(19(30)22(5,6)7)27-20(33-23)26-18(29)21(2,3)4/h9-13,24-25H,8,14-16H2,1-7H3,(H,26,27,29)/t23-/m0/s1
Chemical Name	N-[(5R)-4-(2,2-dimethylpropanoyl)-5-[[2-(ethylamino)ethylsulfonylamino]methyl]-5-phenyl-1,3,4-thiadiazol-2-yl]-2,2-dimethylpropanamide
Synonyms	LY2523355; KF 89617; LY-2523355; KF89617; LY 2523355; KF-89617; Eg5 KinesinRelated Motor Protein Inhibitor
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: ≥ 50 mg/mL (~97.7 mM)
Solubility (In Vivo)	Solubility in Formulation 1: ≥ 2.5 mg/mL (4.89 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 (4.89 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. View More Solubility in Formulation 3: ≥ 2.5 mg/mL (4.89 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.)

Solubility (In Vitro)

DMSO: ≥ 50 mg/mL (~97.7 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.89 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 (4.89 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.

View More

Solubility in Formulation 3: ≥ 2.5 mg/mL (4.89 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	1.9543 mL	9.7714 mL	19.5427 mL
	5 mM	0.3909 mL	1.9543 mL	3.9085 mL
	10 mM	0.1954 mL	0.9771 mL	1.9543 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.

Calculator

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See Example

An example of molarity calculation using the molarity calculator is shown below:
What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?

Enter 350.26 in the Molecular Weight (MW) box
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Click the “Calculate” button
The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

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Dilution Calculator allows you to calculate how to dilute a stock solution of known concentrations. For example, you may Enter C1, C2 & V2 to calculate V1, as detailed below:

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:

Enter 10 into the Concentration (Start) box and choose the correct unit (mM)
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The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box

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Molecular Weight Calculator allows you to calculate the molar mass and elemental composition of a compound, as detailed below:

Note: Chemical formula is case sensitive: C12H18N3O4 c12h18n3o4
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Definitions of molecular mass, molecular weight, molar mass and molar weight:

Molecular mass (or molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.

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The answer appears in the Volume (to add to vial) box

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)

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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.)

% DMSO

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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.

Note： (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.

Clinical Trial Information

NCT Number	Recruitment	interventions	Conditions	Sponsor/Collaborators	Start Date	Phases
NCT01358019	Completed	Drug: LY2523355	Solid Tumors	Kyowa Kirin Co., Ltd.	May 2011	Phase 1
NCT01416389	Completed	Drug: LY2523355 Drug: ixabepilone	Metastatic Breast Cancer	Eli Lilly and Company	May 25, 2018	Phase 2
NCT01214629	Completed	Drug: LY2523355 Drug: pegfilgrastim	Advanced Cancer Metastatic Cancer	Eli Lilly and Company	July 2007	Phase 1
NCT01059643	Completed	Drug: LY2523355 Drug: pegfilgrastim	Ovarian Cancer Prostate Cancer	Eli Lilly and Company	April 2011	Phase 2
NCT01214642	Completed	Drug: pegfilgrastim Drug: LY2523355	Advanced Cancer	Eli Lilly and Company	May 2008	Phase 1

Biological Data

LY2523355 kills cancer cells during mitotic arrest. Mol Cancer Ther (2015) 14 (11): 2463–2472.
Sustained activation of SAC is required for rapid cell death by LY2523355. Mol Cancer Ther (2015) 14 (11): 2463–2472.
Anticancer efficacy of LY2523355 in xenograft tumor models. Mol Cancer Ther (2015) 14 (11): 2463–2472.