| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
Enterovirus A71 (EV-A71)
LY2334737 is a substrate of human carboxylesterase 2 (CES2); instead, CES2 hydrolyzes the prodrug with Michaelis‑Menten parameters: Km = 43 μmol/L, Vmax = 40 nmol/min/mg protein. CES1 and CES3 showed no hydrolytic activity [1]. |
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| ln Vitro |
LY2334737 treatment produced a response in five CES2-expressing cell lines. SK-OV-3 CES2 knockdown cells are less cytotoxic to LY2334737 than to parental cells. The level of CES2 expression was correlated with the drug response of HCT-116 cells transfected with CES2. Bystander investigations reveal that when cells are cocultured with CES2 rather than mock transfectants, LY2334737 statistically significantly inhibits PC-3-GFP growth[1].
LY2334737 is noncytotoxic in vitro unless activated by CES2. In a panel of cell lines, those expressing CES2 (e.g., 2008, HepG‑2, Colo‑205, A498, SK‑OV‑3, MSTO‑211H) showed sensitivity to LY2334737 with EC50 ratios (LY2334737/gemcitabine) ≤ 15, whereas CES2‑negative cells (HCT‑116, HEK293) had ratios ≥ 300 [1]. In HCT‑116 cells stably transfected with CES2, the prodrug EC50 was 0.16 μM (14‑fold more sensitive than mock transfectant, EC50 = 2.33 μM). CES1 transfection did not increase sensitivity. A strong correlation (r² = 0.94) was observed between CES2 transcript levels and LY2334737 cytotoxicity in CES2 transfectants [1]. Knockdown of CES2 in SK‑OV‑3 cells (80‑90% reduction) decreased LY2334737 sensitivity by 4‑fold (EC50 increased from 1.1 to 4.5 μM); addition of the CES2 inhibitor loperamide (10 μM) further reduced sensitivity of parental cells by 2.5‑fold but had little effect on the knockdown. Gemcitabine cytotoxicity was unaffected [1]. Bystander effect: When GFP‑tagged PC‑3 cells (CES2‑negative) were cocultured with CES2‑expressing HCT‑116 transfectants, LY2334737 cytotoxicity to PC‑3 cells was enhanced 7.5‑fold (EC50 from 1.35 to 0.18 μM) compared to coculture with mock transfectants. No bystander effect was seen with gemcitabine or G418 [1]. |
| ln Vivo |
Tumor growth inhibition of CES2 transfectant is higher than that of mock transfectant when 3.2 mg/kg LY2334737 is administered orally to xenograft models once daily for 21 days|1. In luciferase-tagged LM2-4 tumor xenografts, metronomic LY2334737 administration results in increased blood flow. This effect can be easily measured using contrast micro-ultrasound, and it coincides with a relative increase in tumor bioluminescence[3].
LY2334737 (CAS#: 892128-60-8) showed antiviral efficacy in a lethal EV-A71 infection model in 6-day-old BALB/c mice. Oral administration at 0.32 mg/kg (equimolar to 0.25 mg/kg gemcitabine) once daily for 5 doses resulted in 50% survival (vs 0% in PBS control), with lower mean clinical scores (maximum 4 vs 6 in controls). Viral titers in limb muscle tissue were not significantly reduced (0.7 log PFU/mL reduction, p=0.05), but muscle necrosis was mild compared to severe necrosis in controls. [2] In orthotopic human breast cancer (LM2-4luc+) xenografts in SCID mice, daily oral LY2334737 (CAS#: 892128-60-8) at 6 mg/kg significantly inhibited tumor growth (p<0.05) after 3 weeks of treatment. It did not reduce microvessel density; instead it increased intratumoral blood flow at 1 and 3 weeks post-treatment. Combination with metronomic cyclophosphamide caused toxicity. [3] In an orthotopic human ovarian cancer (SKOV3-13) model, daily oral LY2334737 (CAS#: 892128-60-8) at 6 mg/kg significantly prolonged survival compared to vehicle control (p<0.05). Luminescence imaging showed significant antitumor effect. Combination with low-dose cyclophosphamide did not further improve survival. [3] |
| Enzyme Assay |
Human recombinant CES1A1, CES2, and CES3 were expressed and assayed for hydrolysis of p‑nitrophenyl butyrate (PNB) to confirm activity. For LY2334737, [³H]‑labeled prodrug (100 nmol/L) was incubated with each recombinant CES (0.2 μg/μL) in 50 mmol/L Tris‑HCl (pH 7.4) at 37 °C for 2‑18 h. Reactions were terminated with methanol, and gemcitabine formation was quantified by HPLC. Only CES2 hydrolyzed the prodrug. Kinetic parameters for CES2 were determined using 2.5‑250 μmol/L [³H]LY2334737 at 30 min incubation; Km = 43 μmol/L, Vmax = 40 nmol/min/mg [1].
Hydrolysis of 4‑methylumbelliferyl butyrate (4‑MUB) was used to screen CES transfectants and knockdown cells. Cells were lysed in buffer, incubated with 100 μmol/L 4‑MUB for 10 min, and fluorescence (excitation 355 nm, emission 460 nm) was measured. Hydrolysis of PNB was monitored at 405 nm [1]. |
| Cell Assay |
Cell viability was measured using CellTiter 96 AQueous One Solution reagent (MTS). Cells were seeded in 96‑well plates (2,000‑10,000/well) in growth medium with 10% FBS, incubated with serial dilutions of LY2334737 or gemcitabine for 3 or 5 days, then MTS reagent was added and absorbance read at 490 nm. EC50 values were calculated [1].
For bystander studies, PC‑3‑GFP cells (CES2‑negative) were grown alone or cocultured (1:1 ratio) with mock or CES2‑HCT‑116 transfectants in poly‑D‑lysine‑coated clear‑bottom plates for 3 days. Fluorescence (excitation 475 nm, emission 535 nm) was measured using a Cellomics ArrayScan VTi to quantify PC‑3‑GFP viability. EC50 values for gemcitabine, LY2334737, and G418 were determined [1]. CES2 expression was measured by qRT‑PCR (normalized to GAPDH) and Western blotting using anti‑CES2 antibody. Immunohistochemical staining of cell pellets and tumor sections was performed with anti‑CES2 antibody and positive pixel count analysis [1]. |
| Animal Protocol |
For antiviral studies (literature 2): 6-day-old suckling BALB/c mice (n=6-10 per group) were infected intraperitoneally with 2×10⁷ PFU EV-A71. LY2334737 (CAS#: 892128-60-8) was administered by oral gavage at 0.32 mg/kg per mouse (equimolar to 0.25 mg/kg gemcitabine) once daily for 5 doses starting at 1 hpi. PBS was used as control. Body weight, survival, clinical scores (scoring system based on activity, breathing, movement, body weight) were monitored for 14 days. On day 7, limb muscle tissues were collected for viral plaque assay and histology (H&E and IHC). [2]
For metronomic chemotherapy studies (literature 3): Female Balb/cJ mice (non-tumor bearing) received daily oral LY2334737 (CAS#: 892128-60-8) by gavage at doses of 2, 4, 6, 8, 10, 15, 20 mg/kg for up to 28 days. The drug was prepared as a 2.0 mg/mL solution in 0.1 M sodium phosphate buffer (pH 6.0), stored at 4°C in the dark, and diluted as needed. For efficacy studies, female SCID mice bearing orthotopic LM2-4luc+ breast tumors (average size 250 mm³) or intraperitoneal SKOV3-13 ovarian tumors were treated with daily oral LY2334737 at 6 mg/kg. Treatment continued for 2-3 weeks. Blood flow was measured by high-frequency micro-ultrasound functional imaging with contrast agent. Tumor microvessel density was assessed by CD31 immunostaining. [3] |
| ADME/Pharmacokinetics |
LY2334737 (CAS#: 892128-60-8) is an oral prodrug of gemcitabine that is absorbed intact and slowly releases gemcitabine systemically over an extended period, consistent with formation-rate-limited kinetics. It is stable between pH 6 and 8. In vivo, it is hydrolyzed by carboxylesterase 2 (CES2) to release gemcitabine, bypassing rapid deamination by cytidine deaminase. Oral administration results in sustained gemcitabine exposure. [2][3]
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| Toxicity/Toxicokinetics |
In non-tumor-bearing Balb/c mice, daily oral LY2334737 (CAS#: 892128-60-8) at doses of 2, 4, and 6 mg/kg for 28 days caused no significant weight loss (<10%) and no reduction in white blood cell counts, indicating minimal toxicity. Doses above 10 mg/kg (e.g., 15 or 20 mg/kg for 7 days) caused severe weight loss (>20%) and toxicity. In SCID mice bearing LM2-4 tumors, the combination of LY2334737 (8 mg/kg/day) with metronomic cyclophosphamide (bolus 100 mg/kg + 20 mg/kg/day) caused host toxicity (weight loss >10%). In suckling mice, LY2334737 (0.32 mg/kg) showed no significant body weight difference compared to naive controls over 15 days. No effect on circulating endothelial progenitor cells (CEPs) was observed at non-toxic doses (6-8 mg/kg). [3]
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| References |
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| Additional Infomation |
LY2334737 has been used in research trials for the treatment of solid tumors, metastatic tumors, and malignant solid tumors.
The gemcitabine prodrug LY2334737 is an orally effective gemcitabine valproate prodrug. Gemcitabine is a broad-spectrum antimetabolite and deoxycytidine analog with antitumor activity. After administration, the gemcitabine prodrug LY2334737 is hydrolyzed by carboxylesterase 2 (CES2), systematically releasing gemcitabine over a period of time, a process conforming to rate-limiting kinetics. Subsequently, gemcitabine is converted into the active metabolites difluorodeoxycytidine diphosphate and triphosphate (dFdCDP and dFdCTP) by deoxycytidine kinase. dFdCDP inhibits ribonucleotide reductase, thereby reducing the pool of deoxynucleotides available for DNA replication; dFdCTP can be incorporated into DNA, leading to premature termination of DNA replication and ultimately inducing apoptosis. Compared to gemcitabine, this prodrug avoids hydrolysis in intestinal cells and the portal circulation, thus avoiding first-pass metabolism and improving the systemic bioavailability of gemcitabine. Furthermore, sustained release of gemcitabine may enhance efficacy and reduce toxicity. The serine ester hydrolase CES2 is expressed in some tumors, which may promote gemcitabine transformation at tumor sites, thereby enhancing its cytotoxicity. LY2334737 (CAS#: 892128-60-8) is a gemcitabine prodrug designed to improve oral bioavailability and reduce rapid deamination. It has been evaluated in phase I clinical trials for advanced solid tumors. In preclinical studies, metronomic dosing (daily oral) caused antitumor effects without inhibiting systemic vasculogenesis (no reduction in CEPs or microvessel density), suggesting mechanisms independent of anti-angiogenesis, such as increased intratumoral blood flow. [2][3] |
| Molecular Formula |
C17H25F2N3O5
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|---|---|---|
| Molecular Weight |
389.39
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| Exact Mass |
389.176
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| Elemental Analysis |
C, 52.44; H, 6.47; F, 9.76; N, 10.79; O, 20.54
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| CAS # |
892128-60-8
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| Related CAS # |
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| PubChem CID |
11646777
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| Appearance |
White to off-white solid powder
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| LogP |
1.118
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
27
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| Complexity |
620
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| Defined Atom Stereocenter Count |
3
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| SMILES |
FC1([C@H](O)[C@@H](CO)O[C@H]1N1C=CC(NC(=O)C(CCC)CCC)=NC1=O)F
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| InChi Key |
MEOYFIHNRBNEPI-UXIGCNINSA-N
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| InChi Code |
InChI=1S/C17H25F2N3O5/c1-3-5-10(6-4-2)14(25)20-12-7-8-22(16(26)21-12)15-17(18,19)13(24)11(9-23)27-15/h7-8,10-11,13,15,23-24H,3-6,9H2,1-2H3,(H,20,21,25,26)/t11-,13-,15-/m1/s1
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| Chemical Name |
N-[1-[(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-2-oxopyrimidin-4-yl]-2-propylpentanamide
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| Synonyms |
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.42 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 (6.42 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 (6.42 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5681 mL | 12.8406 mL | 25.6812 mL | |
| 5 mM | 0.5136 mL | 2.5681 mL | 5.1362 mL | |
| 10 mM | 0.2568 mL | 1.2841 mL | 2.5681 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.
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.
| NCT Number | Recruitment | interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
| NCT01648764 | Completed | Drug: LY2334737 | Malignant Solid Tumor Solid Tumor |
Eli Lilly and Company | September 2008 | Phase 1 |
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