| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| Other Sizes |
Purity: ≥98%
LY345899 is a novel, potent inhibitor of cytoplasmic and mitochondrial methylenetetrahydrofolate dehydrogenase MTHFD1 and MTHFD2 inhibitor with a Ki of 0.018 μM. LY345899 inhibited MTHFD2 with an IC50 of 663 nM and inhibited MTHFD1 with an IC50 of 96 nM.
| Targets |
LY-345899 is an inhibitor of the cytoplasmic methylene tetrahydrofolate dehydrogenase/cyclohydrolase domain of the human trifunctional enzyme MTHFD1 (also referred to as DC301 in the context of its crystal structure). [2]
|
|---|---|
| ln Vitro |
MTHFD enzyme in the mitochondria and cytoplasm can be inhibited by LY 345899 [4]. In vitro, LY 345899 may impair anchorage independence, whereas in vivo, it may cause tumor development and metastasis, disturb NADPH and redox balance, and hasten cell death under oxidative stress conditions such hypoxia [4].
LY-345899 for 72 hours caused a decrease in the cell viability of colorectal cancer (CRC) cell lines with high MTHFD2 expression (LoVo, SW620, HCT116).[4] Treatment with a low dose of LY-345899 (1 μM) for 14 days statistically significantly reduced colony formation in CRC cells.[4] Treatment with LY-345899 (10 μM) for 24 hours decreased cellular NADPH/NADP+ levels and reduced glutathione/oxidized glutathione (GSH/GSSG) levels in CRC cells.[4] Treatment with LY-345899 increased cellular reactive oxygen species (ROS) levels and caused more cell death in CRC cells, which could be rescued by adding the antioxidant N-acetyl-L-cysteine (NAC).[4] Adding 5 mM formate to the culture media did not rescue the cell apoptosis induced by LY-345899 treatment, suggesting that the cell death is likely due to redox modification rather than a reduction in formate production.[4] Treatment with LY-345899 for 48 hours significantly enhanced cell death in CRC cells under hypoxic conditions or under detached (anchorage-independent) conditions.[4] |
| ln Vivo |
In vivo studies of LY345899 (ip; 5–10 mg/kg; 5 days/week; 4 weeks) showed strong antitumor activity as well as therapeutic activity against CRC [1].
In SW620 cell-based xenograft models, treatment with LY-345899 (at 5 mg/kg or 10 mg/kg per mouse, intraperitoneal injection, 5 days per week) significantly suppressed tumor growth and reduced tumor weight.[4] In patient-derived xenograft (PDX) models from two CRC patients with liver metastasis, treatment with LY-345899 (10 mg/kg per mouse, intraperitoneal injection, 5 days per week) significantly suppressed tumor growth. The mean tumor weight of the vehicle-treated PDX group was 1.83 (0.19) mg versus 0.74 (0.30) mg for the LY-345899-treated group.[4] Immunohistochemical staining of tumors from LY-345899-treated mice showed lower cell proliferation indices and higher cell apoptosis compared to controls.[4] In orthotopic metastasis models where SW620 cells or PDX tissues were implanted into the cecum of nude mice, treatment with LY-345899 significantly reduced the number of mesenteric metastatic nodules.[4] |
| Cell Assay |
For cell viability assays, colorectal cancer cells were treated with LY-345899 for 72 hours. Cell viability was then determined using an MTS assay.[4]
For colony formation assays, CRC cells were treated with a low dose of LY-345899 (1 μM) for 14 days, after which colonies were stained and counted.[4] To measure redox parameters, cells were treated with LY-345899 (10 μM) for 24 hours. Cellular NADPH/NADP+ levels were measured using a commercial assay kit. Cellular ROS levels were measured by flow cytometry using the fluorescent probe CM-H2DCFDA (DCF-DA). The intracellular levels of reduced and oxidized glutathione (GSH/GSSG) were measured using a commercial GSH/GSSG assay kit.[4] For apoptosis assays under stress conditions, cells were treated with LY-345899 for 48 hours under normoxia, hypoxia, or detached conditions. Cell death was measured by Annexin V/PI staining and flow cytometry.[4] To test rescue effects, cells were co-treated with LY-345899 and 5 mM N-acetyl-L-cysteine (NAC) or 5 mM formate, and apoptosis was assessed.[4] |
| Animal Protocol |
Animal/Disease Models: BABL/c nude colorectal cancer (CRC) model based on SW620 or PDX [4]
Doses: 5-10 mg/kg Route of Administration: intraperitoneal (ip) injection; 5-10 mg/kg; 5 days/week; 4 Weekly Experimental Results: demonstrated lower cell proliferation index and higher cell apoptosis. No statistically significant weight loss or other signs of acute or delayed toxicity were demonstrated in mice. For subcutaneous xenograft tumor growth studies, CRC cells (2 x 10^6 cells in phosphate-buffered saline) were subcutaneously injected into the flank of female BALB/c nude mice. Patient-derived xenograft (PDX) tumors from two patients with liver metastasis were also implanted subcutaneously.[4] When tumors became palpable (approximately 100 mm³ in volume), tumor-bearing mice were randomly assigned to treatment or control groups (5 mice per group for PDX models). Mice were treated with LY-345899 via intraperitoneal injection (i.p.) at doses of 5 mg/kg or 10 mg/kg, administered 5 days per week for 4 weeks. Tumor size was measured every 4 days with calipers, and volume was calculated (V = length × width² / 2). Mice were euthanized when they met institutional criteria, and tumors were excised and weighed.[4] For orthotopic metastasis studies, CRC cells or PDX tissues were surgically implanted into the cecum of nude mice (8 mice per group). Mice were treated with LY-345899 (10 mg/kg, i.p., 5 days per week) for 4 weeks. Afterwards, mice were euthanized, intestines were removed, and metastatic nodules on the intestinal wall were counted.[4] |
| Toxicity/Toxicokinetics |
In in vivo studies, no statistically significant weight loss or other signs of acute or delayed toxicity were observed in mice treated with LY-345899 during the treatment period. [4]
|
| References |
|
| Additional Infomation |
LY-345899 (also abbreviated as LY34 in the figure) is a substrate analog inhibitor based on 5,10-methylenetetrahydrofolate (5,10-methylene-THF), an intermediate in the folic acid pathway. [2] The crystal structure of the complex of human MTHFD1 bifunctional methylenetetrahydrofolate dehydrogenase/cyclization hydrolase domain (DC301) with LY-345899 has been resolved, demonstrating how the substrate analog inhibitor binds relative to the NADP cofactor within the enzyme's active site. [2] This structural information was used to compare and contrast the active sites of related enzymes (MTHFD1, MTHFD2, MTHFD2L) for rational drug design, but no functional data (e.g., inhibition constant, cellular efficacy) were provided. LY-345899 was proposed. [2]
|
| Molecular Formula |
C20H21N7O7
|
|---|---|
| Molecular Weight |
471.423443555832
|
| Exact Mass |
471.15
|
| CAS # |
10538-99-5
|
| PubChem CID |
135484014
|
| Appearance |
Light yellow to yellow solid powder
|
| LogP |
-1.6
|
| Hydrogen Bond Donor Count |
6
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
7
|
| Heavy Atom Count |
34
|
| Complexity |
986
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
O=C1N(C2C=CC(C(N[C@H](C(=O)O)CCC(=O)O)=O)=CC=2)CC2CNC3=C(C(NC(N)=N3)=O)N21
|
| InChi Key |
JSNFRYBHBVDHSG-KIYNQFGBSA-N
|
| InChi Code |
InChI=1S/C20H21N7O7/c21-19-24-15-14(17(31)25-19)27-11(7-22-15)8-26(20(27)34)10-3-1-9(2-4-10)16(30)23-12(18(32)33)5-6-13(28)29/h1-4,11-12H,5-8H2,(H,23,30)(H,28,29)(H,32,33)(H4,21,22,24,25,31)/t11?,12-/m0/s1
|
| Chemical Name |
(4-(3-amino-1-hydroxy-9-oxo-5,6,6a,7-tetrahydroimidazo[1,5-f]pteridin-8(9H)-yl)benzoyl)-L-glutamic acid
|
| Synonyms |
LY345899; LY-345899; LY 345899;
|
| 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 (In Vitro) |
DMSO : ~125 mg/mL (~265.16 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.30 mM) (saturation unknown) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
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 (4.41 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. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (4.41 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. Solubility in Formulation 4: ≥ 2.08 mg/mL (4.41 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 corn oil and mix evenly. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1213 mL | 10.6063 mL | 21.2125 mL | |
| 5 mM | 0.4243 mL | 2.1213 mL | 4.2425 mL | |
| 10 mM | 0.2121 mL | 1.0606 mL | 2.1213 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA