LY2510924 (LY-2510924), a small cyclic peptide, is a novel, potent and selective CXCR4 (CXC chemokine receptor 4) antagonist with potential anticancer and anti-inflammatory activity. It inhibits SDF-1 binding to CXCR4 with an IC50 of 0.079 nM. LY2510924 may have antitumor effects since it inhibits the CXC chemokine receptor 4 (CXCR4). Upon subcutaneous administration, the CXCR4 inhibitor LY2510924 binds to the chemokine receptor CXCR4, preventing CXCR4 from binding to its ligand, stromal derived factor-1 (SDF-1), and consequent receptor activation. Tumor cell migration and proliferation may be reduced as a result of this. Chemokine receptor CXCR4, which is part of the G protein-coupled receptor (GPCR) gene family, is upregulated in various tumor cell types and is involved in angiogenesis and chemotaxis.

Physicochemical Properties

Molecular Formula	C62H88N14O10
Molecular Weight	1189.45
Exact Mass	1188.68
Elemental Analysis	C, 62.55; H, 7.37; N, 15.30; O, 14.78
CAS #	1088715-84-7
PubChem CID	25102787
Appearance	White to off-white solid powder
Density	1.3±0.1 g/cm3
Index of Refraction	1.641
LogP	0.58
Hydrogen Bond Donor Count	14
Hydrogen Bond Acceptor Count	13
Rotatable Bond Count	25
Heavy Atom Count	86
Complexity	2200
Defined Atom Stereocenter Count	7
SMILES	C([C@H]1C(NCC(N[C@H](CCC(N[C@H](C(N[C@H](C(N[C@H](C(N[C@@H](C(N1)=O)CCCNC(N)=N)=O)CCCCNC(C)C)=O)CC1C=CC(=CC=1)O)=O)CC1C=CC=CC=1)=O)C(=O)N[C@H](C(=O)N)CCCCNC(C)C)=O)=O)C1C=CC2C=CC=CC=2C=1
InChi Key	IJHWVENTEFSNBC-OFPUNPKKSA-N
InChi Code	InChI=1S/C62H88N14O10/c1-38(2)66-30-12-10-19-46(55(63)80)72-59(84)49-28-29-53(78)71-51(34-40-15-6-5-7-16-40)60(85)76-52(35-41-23-26-45(77)27-24-41)61(86)74-47(20-11-13-31-67-39(3)4)57(82)73-48(21-14-32-68-62(64)65)58(83)75-50(56(81)69-37-54(79)70-49)36-42-22-25-43-17-8-9-18-44(43)33-42/h5-9,15-18,22-27,33,38-39,46-52,66-67,77H,10-14,19-21,28-32,34-37H2,1-4H3,(H2,63,80)(H,69,81)(H,70,79)(H,71,78)(H,72,84)(H,73,82)(H,74,86)(H,75,83)(H,76,85)(H4,64,65,68)/t46-,47-,48+,49+,50-,51-,52+/m0/s1
Chemical Name	(2S,5R,8S,11R,14S,20R)-N-[(2S)-1-amino-1-oxo-6-(propan-2-ylamino)hexan-2-yl]-2-benzyl-11-[3-(diaminomethylideneamino)propyl]-5-[(4-hydroxyphenyl)methyl]-14-(naphthalen-2-ylmethyl)-3,6,9,12,15,18,23-heptaoxo-8-[4-(propan-2-ylamino)butyl]-1,4,7,10,13,16,19-heptazacyclotricosane-20-carboxamide
Synonyms	LY2510924; LY-2510924; LY 2510924; Cyclo[Phe-Tyr-Lys(iPr)-D-Arg-2-Nal-Gly-D-Glu]-Lys(iPr)-NH2
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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.
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

Targets	125I-SDF-1α-CXCR4 ( Ki = 79.7 pM ); 125I-SDF-1α-CXCR4 ( Ki = 49.5 pM )
ln Vitro	LY2510924 selectively inhibits SDF-1-induced GTP binding with a Kb value of 0.38 nM and blocks SDF-1 binding to CXCR4 with an IC50 value of 0.079 nM. LY2510924 inhibits SDF-1-induced cell migration and SDF-1/CXCR4-mediated intracellular signaling in human lymphoma U937 cells that express endogenous CXCR4. The IC50 value of this inhibitor is 0.26 nM. Tumor cells stimulated with SDF-1 show concentration-dependent inhibition of phospho-ERK and phospho-Akt. LY2510924 does not appear to have any agonist activity, according to biochemical and cellular analyses[1]. The main effects of LY2510924 are to reduce stromal cell defense against chemotherapy and to inhibit AML cell proliferation with minimal induction of cell death[2].
ln Vivo	LY2510924 selectively inhibits SDF-1-induced GTP binding with a Kb value of 0.38 nM and blocks SDF-1 binding to CXCR4 with an IC50 value of 0.079 nM. LY2510924 inhibits SDF-1-induced cell migration and SDF-1/CXCR4-mediated intracellular signaling in human lymphoma U937 cells that express endogenous CXCR4. The IC50 value of this inhibitor is 0.26 nM. Tumor cells stimulated with SDF-1 show concentration-dependent inhibition of phospho-ERK and phospho-Akt. LY2510924 does not appear to have any agonist activity, according to biochemical and cellular analyses[1]. The main effects of LY2510924 are to reduce stromal cell defense against chemotherapy and to inhibit AML cell proliferation with minimal induction of cell death[2].
Enzyme Assay	LY2510924 selectively inhibits SDF-1-induced GTP binding with a Kb value of 0.38 nM and blocks SDF-1 binding to CXCR4 with an IC50 value of 0.079 nM. LY2510924 inhibits SDF-1-induced cell migration and SDF-1/CXCR4-mediated intracellular signaling in human lymphoma U937 cells that express endogenous CXCR4. The IC50 value of this inhibitor is 0.26 nM. Tumor cells stimulated with SDF-1 show concentration-dependent inhibition of phospho-ERK and phospho-Akt. LY2510924 does not appear to have any agonist activity, according to biochemical and cellular analyses[1]. The main effects of LY2510924 are to reduce stromal cell defense against chemotherapy and to inhibit AML cell proliferation with minimal induction of cell death[2].
Cell Assay	After harvesting U937 cells, they are gently rinsed once with chemotaxis assay buffer, which is made with 1× RPMI medium that contains 10 mmol/L HEPES, pH 7.5, and 0.3% BSA. At a density of 5 × 106 cells/mL, the cells are then resuspended in assay buffer. A 96-well ChemoTx plate is used for the assay. Usually, the lower chamber was filled with 30 μL of SDF-1α (10 ng/mL) prepared in 1× chemotaxis buffer, while the upper chamber was plated with 50 μL of cell mixture, either with or without LY2510924. The plate is then incubated at 37°C for 2.5 hours. After the incubation period, the lower chamber is filled with 5 μL of CellTiter 96 AQ. The plate is then incubated at 37°C for 60 minutes. By measuring the absorbance at 492 nm, the migrated cells can be identified.
Animal Protocol	Mice: MDA-MB-231 cells are injected intravenously into female SCID mice, and they receive subcutaneous injections of either vehicle (1×PBS) or LY2510924 (3 mg/kg) formulated in 1×PBS. The animals in Groups 1 and 2 are treated with either a vehicle or 3 mg/kg of LY2510924 twice a day for several days, starting one day prior to the injection of tumor cells. Treatment for Group 3 animals starts one day after tumor cell injection and lasts for 13 days, with each animal receiving 3 mg/kg of LY2510924 15 twice daily. Lung lobes are visible in histologic sections after lung tissues are preserved in 10% neutral-buffered formalin for at least 24 hours following treatment[1].
References	[1]. Identification of LY2510924, a novel cyclic peptide CXCR4 antagonist that exhibits antitumor activities in solid tumor and breast cancer metastatic models. Mol Cancer Ther. 2015 Feb;14(2):480-90. [2]. Antileukemia activity of the novel peptidic CXCR4 antagonist LY2510924 as monotherapy and in combination with chemotherapy. Blood. 2015 Jul 9;126(2):222-32.

Solubility Data

Solubility (In Vitro)

DMSO: ≥ 125 mg/mL (~105.1 mM) Water: ~100 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (1.75 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 20.8 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: ≥ 2.08 mg/mL (1.75 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 20.8 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: ≥ 2.08 mg/mL (1.75 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 20.8 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	0.8407 mL	4.2036 mL	8.4072 mL
	5 mM	0.1681 mL	0.8407 mL	1.6814 mL
	10 mM	0.0841 mL	0.4204 mL	0.8407 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.