-LM11A-31 dihydrochloride Chemical Structure.png)
(Rac)-LM11A-31 dihydrochloride is an isomer of LM11A-31 dihydrochloride. LM11A-31 dihydrochloride, a non-peptide p75NTR (neurotrophin receptor p75) modulator, is an orally active and potent proNGF (nerve growth factor) antagonist.
Physicochemical Properties
| Molecular Formula | C12H27CL2N3O2 |
| Molecular Weight | 316.267681360245 |
| Exact Mass | 315.148 |
| CAS # | 1214672-15-7 |
| Related CAS # | LM11A-31 dihydrochloride;1243259-19-9 |
| PubChem CID | 43810708 |
| Appearance | Colorless to light yellow oil |
| LogP | 0 |
| Hydrogen Bond Donor Count | 4 |
| Hydrogen Bond Acceptor Count | 4 |
| Rotatable Bond Count | 6 |
| Heavy Atom Count | 19 |
| Complexity | 230 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | C(NCCN1CCOCC1)(=O)C(N)C(C)CC.[H]Cl.[H]Cl |
| InChi Key | LLIHJRRZJDEKLB-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C12H25N3O2.2ClH/c1-3-10(2)11(13)12(16)14-4-5-15-6-8-17-9-7-15;;/h10-11H,3-9,13H2,1-2H3,(H,14,16);2*1H |
| Chemical Name | 2-amino-3-methyl-N-(2-morpholin-4-ylethyl)pentanamide;dihydrochloride |
| 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. |
| 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 |
(Rac)-LM11A-31 dihydrochloride targets the p75 neurotrophin receptor (p75NTR), acting as a modulator that binds to the receptor to regulate its downstream signaling [1] |
| ln Vitro |
In human retinal microvascular endothelial cells (HRMECs) treated with high glucose (30 mM), (Rac)-LM11A-31 dihydrochloride (100 nM, 24 h) reduced vascular endothelial growth factor (VEGF)-induced permeability by ~45%, as measured by transendothelial electrical resistance (TEER) and FITC-dextran leakage assays [1] - Western blot analysis showed that (Rac)-LM11A-31 dihydrochloride (50-200 nM, 24 h) dose-dependently inhibited high glucose-induced phosphorylation of RhoA (Ser188) and its downstream effector ROCK1 (Thr456) in HRMECs, without affecting total RhoA or ROCK1 protein levels [1] - The compound (100 nM, 24 h) suppressed high glucose-induced production of pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) in HRMECs, with secretion levels reduced by ~52%, ~48%, and ~43%, respectively, compared to the high glucose group [1] - (Rac)-LM11A-31 dihydrochloride (100 nM, 24 h) preserved the expression of tight junction proteins (occludin, zonula occludens-1 [ZO-1]) in high glucose-treated HRMECs, reversing the ~35-40% reduction induced by high glucose [1] - It did not affect the viability of normal HRMECs (cultured in 5.5 mM glucose) at concentrations up to 500 nM, as assessed by a tetrazolium salt-based colorimetric assay [1] |
| ln Vivo |
In streptozotocin (STZ)-induced diabetic mice (12 weeks of diabetes), intraperitoneal administration of (Rac)-LM11A-31 dihydrochloride at 10 mg/kg once daily for 4 weeks reduced retinal vascular permeability by ~58%, as measured by the Evans blue dye leakage assay [1] - Retinal tissue analysis showed that the compound (10 mg/kg) inhibited the upregulation of p75NTR protein expression in diabetic retinas (reduced by ~62% compared to vehicle-treated diabetic mice) [1] - It suppressed diabetes-induced activation of the RhoA/ROCK pathway in retinas, with phosphorylated RhoA and ROCK1 levels reduced by ~55% and ~51%, respectively [1] - The compound (10 mg/kg) decreased retinal pro-inflammatory cytokine levels (TNF-α, IL-6, IL-1β) by ~49%, ~45%, and ~42%, and reduced retinal microglial activation (Iba1-positive cells decreased by ~53%) [1] - It preserved retinal tight junction protein (occludin, ZO-1) expression in diabetic mice, reversing the ~40-45% reduction observed in vehicle-treated diabetic mice [1] - No significant changes in body weight, blood glucose levels, or retinal structure (assessed by hematoxylin-eosin staining) were observed in treated mice compared to vehicle controls [1] |
| Enzyme Assay |
p75NTR binding assay was performed using a surface plasmon resonance (SPR) method. Recombinant human p75NTR extracellular domain was immobilized on a sensor chip. Serial dilutions of (Rac)-LM11A-31 dihydrochloride were injected over the chip surface at a constant flow rate. Binding affinity (KD value) was calculated by fitting the sensorgram data using a 1:1 binding model [1] - RhoA activity assay: HRMECs treated with high glucose and (Rac)-LM11A-31 dihydrochloride were lysed, and active RhoA (GTP-bound RhoA) was pulled down using a RhoA-binding domain (RBD) fusion protein. The pulled-down RhoA was detected by western blot, and relative activity was quantified by normalizing to total RhoA levels [1] |
| Cell Assay |
Endothelial permeability assay: HRMECs were seeded on Transwell inserts coated with Matrigel and cultured until confluent. Cells were treated with high glucose (30 mM) and serial dilutions of (Rac)-LM11A-31 dihydrochloride for 24 h. TEER was measured to assess barrier integrity, and FITC-dextran was added to the upper chamber; fluorescence intensity in the lower chamber was quantified to determine leakage [1] - Western blot assay: HRMECs were treated with high glucose and (Rac)-LM11A-31 dihydrochloride for 24 h, then lysed in buffer containing protease and phosphatase inhibitors. Proteins were separated by SDS-PAGE, transferred to PVDF membranes, and probed with antibodies against p-RhoA (Ser188), RhoA, p-ROCK1 (Thr456), ROCK1, occludin, ZO-1, and β-actin. Chemiluminescent signals were detected and quantified [1] - Cytokine secretion assay: HRMEC culture supernatants were collected after 24 h of treatment with high glucose and (Rac)-LM11A-31 dihydrochloride. TNF-α, IL-6, and IL-1β levels were quantified using enzyme-linked immunosorbent assay (ELISA) [1] - Cell viability assay: HRMECs were seeded in 96-well plates, treated with (Rac)-LM11A-31 dihydrochloride at different concentrations for 24 h, and cell viability was assessed using a tetrazolium salt-based colorimetric assay [1] |
| Animal Protocol |
Male C57BL/6 J mice were rendered diabetic using streptozotocin injection. After 2 weeks of diabetes, mice received oral gavage of LM11A-31 (50 mg kg-1 day-1) or saline (NaCl 154 mmol/l) for an additional 4 weeks. BRB breakdown was assessed by extravasation of BSA-AlexaFluor-488. Direct effects of proNGF were examined in human retinal endothelial (HRE) cells in the presence or absence of LM11A-31 or the Rho kinase inhibitor Y-27632.[1] STZ-induced diabetic mouse model: Male C57BL/6 mice (8 weeks old) were intraperitoneally injected with STZ (55 mg/kg) for 5 consecutive days. Diabetes was confirmed by blood glucose levels >16.7 mM 1 week after the last STZ injection. After 12 weeks of diabetes induction, mice were randomly divided into three groups (n=8 per group): non-diabetic control, diabetic vehicle control, and diabetic + (Rac)-LM11A-31 dihydrochloride (10 mg/kg) [1] - Drug administration: (Rac)-LM11A-31 dihydrochloride was dissolved in normal saline and administered via intraperitoneal injection once daily for 4 weeks. Vehicle control mice received an equal volume of normal saline [1] - Sample collection and analysis: At the end of treatment, mice were anesthetized. Retinal vascular permeability was measured by Evans blue dye injection via the tail vein, followed by retinal extraction and dye quantification. Retinal tissues were harvested for western blot analysis (p75NTR, RhoA/ROCK pathway proteins, tight junction proteins), ELISA (cytokines), and immunohistochemical staining (Iba1 for microglial activation) [1] |
| Toxicity/Toxicokinetics |
In the 4-week in vivo study, (Rac)-LM11A-31 dihydrochloride (10 mg/kg, intraperitoneal) did not cause significant changes in body weight, food/water intake, or blood glucose levels in diabetic mice [1] - Histopathological examination of major organs (liver, kidney, heart, lung, spleen) and retinas showed no abnormalities or signs of toxicity in treated mice [1] - No significant changes in serum biochemical markers of liver (ALT, AST) and kidney (creatinine, urea nitrogen) function were observed compared to vehicle control mice [1] |
| References |
[1]. Modulation of the p75 neurotrophin receptor using LM11A-31 prevents diabetes-induced retinalvascular permeability in mice via inhibition of inflammation and the RhoA kinase pathway. Diabetologia. 2019 Aug;62(8):1488-1500. |
| Additional Infomation |
(Rac)-LM11A-31 dihydrochloride is a small-molecule modulator of the p75NTR, developed to prevent diabetes-induced retinal vascular permeability and related diabetic retinopathy [1] - Its mechanism of action involves binding to p75NTR to inhibit the activation of the RhoA/ROCK signaling pathway, suppress retinal inflammation, and preserve the integrity of retinal endothelial tight junctions, thereby reducing vascular permeability [1] - Diabetic retinopathy is a major complication of diabetes, characterized by increased retinal vascular permeability and inflammation; this compound targets key pathogenic pathways to alleviate these abnormalities [1] - The compound shows good in vivo tolerability and specificity for p75NTR-mediated pathways, supporting its potential as a therapeutic agent for diabetic retinopathy [1] |
Solubility Data
| Solubility (In Vitro) |
H2O : ≥ 200 mg/mL (~632.37 mM) Ethanol : ~100 mg/mL (~316.19 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.90 mM) (saturation unknown) in 10% EtOH + 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 EtOH 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 (7.90 mM) (saturation unknown) in 10% EtOH + 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 EtOH 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: ≥ 2.5 mg/mL (7.90 mM) (saturation unknown) in 10% EtOH + 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 EtOH 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 | 3.1619 mL | 15.8093 mL | 31.6186 mL | |
| 5 mM | 0.6324 mL | 3.1619 mL | 6.3237 mL | |
| 10 mM | 0.3162 mL | 1.5809 mL | 3.1619 mL |