| 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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| 500mg |
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| Other Sizes |
| Targets |
Tubulin (colchicine binding site; inhibits tubulin polymerization with IC50 = 2.0 ± 0.5 μM in in vitro tubulin polymerization assay; competes with colchicine for binding: at 30 μM, inhibits [³H]colchicine binding by 93.7% ± 0.7, similar to colchicine itself) [1].
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| ln Vitro |
LP-261 induced G2/M cell cycle arrest in a dose-dependent manner in MCF-7, Jurkat, U266, NCI-H522, SW-620, H23, BxPC-3, and PC3 cell lines with IC50 values ranging from 0.01 to 0.12 μM across these cell lines (specific values: MCF-7 0.01 μM, U266 0.02 μM, H522 0.01 μM, Jurkat 0.02 μM, SW-620 0.05 μM, H23 0.38 μM, BxPC-3 0.01 μM, PC-3 0.02 μM) [1].
LP-261 inhibited cell proliferation in Jurkat, SW-620, A2780, HeLa, and NCI-ADR (multidrug resistant, MDR1) cell lines with IC50 values of 0.09, 0.20, 0.20, 0.21, and 0.30 μM, respectively; it was active against the paclitaxel-resistant NCI-ADR MDR1 cell line, suggesting it is not a substrate for P-glycoprotein-mediated multidrug resistance [1]. In an in vitro tubulin polymerization assay, LP-261 inhibited microtubule formation with an IC50 of 2.0 ± 0.5 μM, comparable to colchicine (2.0 μM) and vincristine (2.5 μM) [1]. LP-261 competitively inhibited [³H]colchicine binding to tubulin: at 30 μM, it inhibited binding by 93.7% ± 0.7, similar to colchicine (93.5%) [1]. |
| ln Vivo |
In a human NCI-H522 non-small-cell lung cancer xenograft model in athymic NCR-nu mice, oral administration of LP-261 at 50 mg/kg twice daily for 28 days resulted in 96% reduction in mean tumor volume (mean tumor volume 130 ± 190 mm³ vs. 3769 ± 1636 mm³ in vehicle controls, P<0.001). At 15 mg/kg b.i.d., tumor growth inhibition was 41%. No significant changes in body weights were observed in the LP-261 treated groups compared to vehicle controls [1].
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| Enzyme Assay |
In vitro tubulin polymerization assay: The assay was conducted using a tubulin polymerization assay kit according to manufacturer's recommendations. Tubulin polymerization was detected by measuring absorbance at 340 nm once every minute over 60 minutes during incubation at 37°C. The final DMSO concentration in each assay well was 1%, and the final concentration series of each test compound was 10, 5, 2.5, 1.25, and 0.6 μM. Colchicine and vincristine were used as reference compounds [1].
Colchicine competition binding assay: [³H]colchicine competition binding assay was performed using biotinylated tubulin. Compounds were assessed at 30 μM for their ability to inhibit [³H]colchicine binding. Inhibition percentages were calculated relative to controls. Vincristine was used as a negative control (did not inhibit colchicine binding) [1]. Computational modeling: The tubulin structure from PDB code 1SA0 (resolved to 3.5 Å) was used. A fragment-based grand canonical Monte Carlo simulation approach was used to identify high-affinity fragment poses. The indole fragment was found to have high affinity poses involving hydrogen bonding with Glu71 and Asp68. LP-261 was assembled by linking fragments (indole, acetophenone, 2-methoxypyridine, N-methylsulfonamide) in their computed positions, allowing some flexibility. The putative binding pose spans both the GTP and colchicine binding sites, with the sulfonamide NH making a hydrogen bond with Glu182 [1]. |
| Cell Assay |
Cell viability assay (WST-1): Jurkat and HeLa cells were cultured in RPMI and EMEM media, respectively, containing 0.5% fetal calf serum. Cells were plated in 96-well format (100 μL/well). Test compounds were added at concentrations ranging from 1 nM to 10 μM for 48-72 h. Cell viability was determined using the WST-1 colorimetric mitochondrial reduction assay: 10 μL of WST-1 reagent was added per well for 1-4 h, then absorbance was read at 450 nm with baseline at 600 nm. Concentration-response curves and IC50 values were calculated using GraphPAD Prism [1].
Cell cycle assay (G2/M block): MCF-7, Jurkat, U266, SW620, NCI-H522, NCI-H23, BxPC-3, and PC3 cells were cultured in appropriate media with 10% FBS. Cells were plated in 24-well plates at 2×10⁵ cells/mL (1 mL/well) and incubated overnight. FBS concentration was reduced to 0.5% for an additional 30 min. Cells were then incubated with various concentrations of test compound for 24 h. Cells were fixed in 80% ethanol for 30 min at room temperature, and nuclear DNA was stained with propidium iodide (PI)/RNase staining buffer. FACS analysis was performed on a FACSalibur system with ~2000 events counted per condition. G1 and G2/M cell populations were measured and reported as percentage of total cell count [1]. Cell proliferation assay: Jurkat, SW-620, A2780, HeLa, and NCI-ADR (MDR1) cells were tested. IC50 values were determined as described for cell viability assay (details not fully specified but similar to WST-1 method) [1]. |
| Animal Protocol |
For in vivo efficacy studies, cultured NCI-H522 tumors (30-40 mm³) were implanted subcutaneously into the thighs of athymic NCR-nu mice and allowed to grow to approximately 150 mm³. Mice were assigned to groups by matching median tumor volume (range 120-162 mm³). The study lasted 28 days, and tumor volume was measured every third day. LP-261 was formulated as a free base using DMSO/propylene glycol/Tween 80/PBS (5:15:2:18 by volume) at 10 mg/mL. It was administered orally twice daily at doses of 15 and 50 mg/kg. Vehicle was administered as the untreated control. Animals with tumors exceeding 4000 mm³ or exhibiting excessive morbidity were sacrificed immediately. Tumor volume for each animal was fitted to exponential growth function Vt = V0 e^(Kt) where Vt is tumor volume at time t, V0 is starting tumor volume, and K is growth rate constant determined from fitting using Prism software [1].
For rat pharmacokinetic studies, LP-261 was administered by oral gavage at 4 mg/kg as a single compound, formulated in a solution of 3:1 polyethylene glycol 400 and 100 mM citrate buffer pH 3. Blood samples were collected at various time points and plasma concentrations were determined by LC-MS/MS after protein precipitation with acetonitrile. Recovery was >85% and lower limit of quantitation was 0.005 μg/mL [1]. |
| ADME/Pharmacokinetics |
Pharmacokinetic parameters of LP-261 in rats after intravenous (2 mg/kg) and oral (4 mg/kg) administration: iv: AUC = 3.7 ± 1.6 μg·h/mL, C0 = 4.9 ± 3.4 μg/mL, half-life = 1.4 ± 0.2 h, CL = 0.67 ± 0.43 L/h/kg, Vss = 1.25 ± 1.13 L/kg. Oral: AUC = 5.9 ± 1.9 μg·h/mL, Tmax = 2.0 h, Cmax = 1.14 ± 0.43 μg/mL, half-life = 1.4 ± 0.2 h, CL = 0.73 ± 0.31 L/h/kg, oral bioavailability = 80% [1].
In rat cassette PK studies (compounds dosed at 4 mg/kg each in cassettes of three), analogues with the phenol group replaced by amide surrogates (including compounds 59, 61, 68, 72, 74) showed improved oral bioavailability ranging from 46% to 66% [1]. The original phenolic lead 7 had only 24% oral bioavailability in rats when dosed at 20 mg/kg in ethanol/propylene glycol/10% Tween 80 in PBS (1:4:5), with extensive glucuronidation observed by LC-MS analysis of plasma [1]. |
| Toxicity/Toxicokinetics |
In rat pharmacokinetic studies, LP-261 was well tolerated with no reported adverse effects at the doses tested. Systemic clearance rate was 0.67 ± 0.43 L/h/kg, which is approximately 5-fold lower than hepatic blood flow in rats (∼3.3 L/h/kg), indicating insignificant hepatic extraction. Volume of distribution (Vss) was 1.25 ± 1.13 L/kg, approximately twice the body water of the rat (0.6 L/kg), suggesting tissue binding. The terminal half-life was 1.4 ± 0.2 h, indicating moderate rate of elimination [1].
In the xenograft efficacy study, no significant changes in body weights were observed for the LP-261 treated groups compared to vehicle treated group, indicating good tolerability at doses up to 50 mg/kg b.i.d. for 28 days. The cisplatin-treated group initially exhibited weight loss that fully recovered by the end of the study [1]. No other toxicity data (e.g., LD50, hematotoxicity, organ toxicity) are reported [1]. |
| References | |
| Additional Infomation |
LP-261 is a potent antimitotic agent that induces G2/M cell cycle arrest and inhibits tubulin polymerization by competing with colchicine at the colchicine binding site on β-tubulin. It is a colchicine-competitive inhibitor of tubulin polymerization, as demonstrated by the [³H]colchicine competition binding assay (93.7% inhibition at 30 μM). The proposed binding mode from computational modeling shows the indole NH hydrogen bonding to Glu71 or Asp68, and the sulfonamide NH hydrogen bonding to Glu182, spanning both the GTP and colchicine binding sites [1].
The compound is not a substrate for P-glycoprotein, as it remained active against the paclitaxel-resistant NCI-ADR MDR1 cell line (IC50 = 0.30 μM), suggesting it will not be subject to multidrug resistance mechanisms [1]. LP-261 was selected for further preclinical evaluation as a promising novel oral anticancer therapy based on its efficacy in a broad array of tumor cell lines, excellent oral bioavailability (80% in rats), and significant antitumor activity in an NCI-H522 xenograft model (96% tumor growth inhibition at 50 mg/kg b.i.d.) [1]. The chemical name of LP-261 is N-[3-(1H-Indol-4-yl)-5-(2-methoxypyridine-4-carbonyl)phenyl]methanesulfonamide. Its structure was confirmed by NMR and HRMS (calcd for C22H19N3O4S, 421.1096, found 421.1093) [1]. |
| Molecular Formula |
C19H15N3O
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|---|---|
| Molecular Weight |
301.3419
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| Exact Mass |
301.122
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| Elemental Analysis |
C, 75.73; H, 5.02; N, 13.94; O, 5.31
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| CAS # |
915410-98-9
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| Related CAS # |
915413-40-0 (HCl);915410-98-9;
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| PubChem CID |
15604188
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| Appearance |
Solid powder
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| LogP |
4.752
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
23
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| Complexity |
391
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| Defined Atom Stereocenter Count |
0
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| SMILES |
N1C2C(=C(C3=CC(NC4=CC=CN=C4)=CC(O)=C3)C=CC=2)C=C1
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| InChi Key |
OSDZVBKVJAPGEB-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H15N3O/c23-16-10-13(17-4-1-5-19-18(17)6-8-21-19)9-15(11-16)22-14-3-2-7-20-12-14/h1-12,21-23H
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| Chemical Name |
3-(1H-indol-4-yl)-5-(pyridin-3-ylamino)phenol
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| Synonyms |
OUN10989; OUN-10989; OUN 10989;
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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 |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.3185 mL | 16.5926 mL | 33.1851 mL | |
| 5 mM | 0.6637 mL | 3.3185 mL | 6.6370 mL | |
| 10 mM | 0.3319 mL | 1.6593 mL | 3.3185 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.
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