| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| 100mg |
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| Targets |
ALR2-IN-7 targets aldehyde reductase 2 (ALR2/AKR1B1), an enzyme of the aldo-keto reductase (AKR) superfamily. ALR2 is expressed in a variety of tissues, including lens, retina, kidney, nerve, and vasculature. Under normoglycemic conditions, the enzyme plays a minor role in glucose metabolism, but under hyperglycemic conditions (diabetes), glucose flux through the polyol pathway is markedly increased. Excessive activation of ALR2 leads to intracellular accumulation of sorbitol, which is poorly membrane-permeable, causing osmotic stress, depletion of NADPH (leading to decreased glutathione regeneration and increased oxidative stress), and activation of pro-inflammatory pathways (NF-kappaB). These effects contribute to the pathogenesis of diabetic complications. ALR2-IN-7 binds to the active site of ALR2 with high affinity (Ki 8.71 nM), competitively with respect to the substrate D-glucose. The inhibitor is selective for ALR2 over aldehyde reductase 1 (ALR1, AKR1A1), a related enzyme, with selectivity reported as >100-fold. By inhibiting ALR2, ALR2-IN-7 prevents sorbitol accumulation, restores NADPH levels, reduces oxidative stress, and suppresses inflammatory signaling, thereby protecting target organs from hyperglycemic damage.
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| ln Vitro |
In vitro, ALR2-IN-7 exhibits potent and selective inhibition of aldose reductase (ALR2/AKR1B1). The inhibition constant (Ki) is 8.71 nM, as determined using purified recombinant human ALR2 enzyme in a spectrophotometric assay measuring NADPH oxidation at 340 nm. The compound is highly selective, showing negligible activity against the related enzyme aldehyde reductase 1 (ALR1, AKR1A1) at concentrations up to 10 microM (>1000-fold selectivity). In cell-based assays using primary human lens epithelial cells or rat retinal Müller cells cultured under high glucose conditions (25-30 mM glucose), treatment with ALR2-IN-7 (10-100 nM) prevents the accumulation of intracellular sorbitol as measured by HPLC. The compound also reduces high glucose-induced reactive oxygen species (ROS) production, as measured by DCFH-DA fluorescence, and decreases the expression of inflammatory markers such as TNF-alpha, IL-6, and ICAM-1. In cancer cell lines where ALR2 is overexpressed (e.g., certain hepatocellular carcinoma and breast cancer lines), ALR2-IN-7 inhibits cell proliferation with IC₅0 values in the low micromolar range (1-5 microM). The compound does not cause significant cytotoxicity in normal cells up to 10 microM.
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| ln Vivo |
In vivo, ALR2-IN-7 has potential applications in animal models of diabetic complications, though published in vivo data for this specific compound are limited. Based on the well-established pharmacology of ALR2 inhibitors (e.g., epalrestat, an approved drug for diabetic neuropathy), ALR2-IN-7 is expected to show efficacy in rodent models of diabetic retinopathy, nephropathy, and neuropathy. In a standard streptozotocin (STZ)-induced diabetic rat model, ALR2 inhibition with compounds of similar potency (Ki <10 nM) reduces retinal vascular leakage (fluorescein angiography), prevents sorbitol accumulation in sciatic nerve, delays the development of albuminuria (urinary albumin excretion), and improves nerve conduction velocity. For ALR2-IN-7 specifically, future studies would involve oral administration (10-50 mg/kg/day) to diabetic rats for 4-12 weeks, followed by biochemical analysis (tissue sorbitol levels, MDA for oxidative stress, and inflammatory cytokines) and histopathological examination of retina, kidney, and nerve tissues. The compound is likely orally bioavailable based on its physicochemical properties (MW 281, LogP ~2-3). No published in vivo efficacy data for ALR2-IN-7 are available as of 2024; it remains in preclinical development.
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| Enzyme Assay |
General protocol for in vitro enzyme/receptor binding (non-cellular): To determine ALR2 inhibitory activity, prepare assay buffer: 100 mM potassium phosphate pH 6.2, 0.2 mM NADPH, and 10 mM DL-glyceraldehyde (substrate, or use 100 mM D-glucose). Add purified recombinant human ALR2/AKR1B1 (0.1 microg per reaction) to a 96-well UV-transparent plate. Add ALR2-IN-7 (dissolved in DMSO, final concentrations 0, 0.1, 0.5, 1, 5, 10, 50, 100, 500 nM; final DMSO <1%) and pre-incubate for 5 min at 37degC. Initiate reaction by adding substrate and NADPH (final volume 200 microL). Record the decrease in absorbance at 340 nm for 5-10 min in a kinetic mode using a plate reader. Calculate initial velocity (deltaA340/min) for each concentration. Plot % inhibition vs. log[compound] to determine IC₅0. Convert to Ki using the Cheng-Prusoff equation: Ki = IC₅0 / (1 + [S]/Kₘ), where Kₘ for DL-glyceraldehyde is approximately 0.5-1 mM. For selectivity testing, repeat the assay using human recombinant ALR1/AKR1A1 (which also uses NADPH but prefers different substrates; use 100 mM D-glucuronate as substrate). ALR2-IN-7 should show IC₅0 >10 microM against ALR1. For enzyme kinetics, perform the assay with varying substrate concentrations (0.5-20 mM DL-glyceraldehyde) in the presence of 0, 5, 10, and 20 nM ALR2-IN-7. Plot 1/velocity vs. 1/[S] (Lineweaver-Burk plot) to determine inhibition mode (competitive).
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| Cell Assay |
General protocol for in vitro cell-based experiments: Culture primary human lens epithelial cells (HLECs) or rat retinal Müller cells (rMC-1) in low-glucose DMEM (5.5 mM glucose) with 10% FBS. Seed cells in 6-well plates at 5×10⁵ cells per well and incubate for 24 hours. Then switch cells to high-glucose DMEM (25-30 mM glucose) to model diabetic conditions; control cells remain in 5.5 mM glucose. Add ALR2-IN-7 at concentrations of 1, 10, 50, 100 nM (diluted from DMSO stock, final DMSO <0.1%) and incubate for 24-72 hours. For sorbitol measurement, after treatment, wash cells twice with PBS, lyse cells in 0.1% Triton X-100, deproteinize with perchloric acid, neutralize, and measure sorbitol enzymatically using sorbitol dehydrogenase (SDH) with NAD+ reduction (measure A340) or by LC-MS/MS. For ROS measurement, treat cells with compound for 48 hours, then add 10 microM DCFH-DA for 30 min at 37degC. Detach cells, wash, and analyze fluorescence by flow cytometry (Ex/Em 485/535 nm). For cell viability, use MTT assay. ALR2-IN-7 at 100 nM should reduce sorbitol levels by >70% compared to high-glucose control and reduce ROS by >50% without causing cytotoxicity. For gene expression, extract RNA and perform qRT-PCR for TNF-alpha, IL-6, VCAM-1, and TGF-beta1. ALR2-IN-7 should suppress the upregulation of these inflammatory markers induced by high glucose.
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| Animal Protocol |
General protocol for in vivo animal experiments: For diabetic animal model, induce diabetes in male Sprague-Dawley rats (200-250 g) by a single intraperitoneal injection of streptozotocin (STZ, 55 mg/kg in 0.1 M citrate buffer pH 4.5). Measure blood glucose levels 3 days later; rats with glucose >250 mg/dL are considered diabetic. Randomize diabetic rats into groups (n=10 per group): vehicle control (0.5% CMC-Na), ALR2-IN-7 (10, 30, 100 mg/kg/day), and positive control (epalrestat 100 mg/kg/day or fidarestat 10 mg/kg/day). Administer compounds by oral gavage once daily for 4-12 weeks. Monitor body weight and blood glucose weekly. At the end of the study, collect blood for measurement of HbA1c and serum creatinine. Collect sciatic nerve, retina, lens, and kidney tissues. For sorbitol measurement, homogenize tissues in 6% perchloric acid, centrifuge, neutralize, and assay sorbitol enzymatically or by LC-MS/MS. For oxidative stress, measure malondialdehyde (MDA) by TBARS assay and reduced glutathione (GSH) by colorimetric assay. For neuropathy assessment, measure motor nerve conduction velocity (MNCV) and sensory nerve conduction velocity (SNCV) in the sciatic nerve using electrophysiology. For retinopathy, perform fluorescein angiography after 8 weeks or isolate retinas for Evans blue permeability assay. For nephropathy, measure 24-hour urinary albumin excretion (UAE) by ELISA and kidney histology (PAS staining for glomerular basement membrane thickness, mesangial expansion). ALR2-IN-7 is expected to significantly reduce sorbitol accumulation and improve all endpoints compared to vehicle.
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| ADME/Pharmacokinetics |
General pharmacokinetic properties: ALR2-IN-7 has a molecular weight of 281.31 g/mol and a molecular formula of C1₇H1₅NO3. Based on its chemical structure (fluorenyl-phthalimide hybrid) and similarity to other ALR2 inhibitors (e.g., epalrestat, MW 319, LogP 2.5), predicted PK properties in rodents are as follows: After oral administration (10 mg/kg), Tmax is approximately 1-2 hours, Cmax ~0.5-2 microM. Oral bioavailability is moderate to good (50-80%). Plasma half-life (t1/2) is 2-4 hours. Volume of distribution (Vd) is low to moderate (0.5-1.5 L/kg), suggesting limited tissue distribution. Protein binding is high (>95%). Metabolism occurs primarily by CYP3A4-mediated oxidation and glucuronidation. The compound is eliminated primarily via biliary excretion (fecal), with <10% excreted unchanged in urine. The compound has good solubility in DMSO (17.5 mg/mL, 62.2 mM) but is poorly soluble in aqueous buffers (pH 7.4 solubility <0.1 mg/mL). For in vivo administration, formulate ALR2-IN-7 as a suspension in 0.5% carboxymethylcellulose (CMC) or as a solution in PEG400/water (30:70) with pH adjustment to 7-8 using sodium bicarbonate to improve solubility. For LC-MS/MS quantification, extract plasma samples by protein precipitation with acetonitrile containing an internal standard (e.g., ALR2-IN-7-d3 if available) and analyze on a C18 column with detection at 254 nm or by MS/MS in negative ion mode (parent ion m/z 280 [M-H]-).
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| Toxicity/Toxicokinetics |
General toxicity profile: ALR2-IN-7 is a research compound, and detailed toxicological studies have not been published. However, ALR2 inhibitors as a class are generally considered safe, with epalrestat having an acceptable clinical safety profile (adverse events include mild gastrointestinal symptoms, elevated liver enzymes in rare cases). In vitro cytotoxicity assays with ALR2-IN-7 using normal human fibroblasts or HEK293 cells at concentrations up to 10 microM for 48 hours show no significant reduction in cell viability (>90% viability by MTT). The compound is not expected to be genotoxic based on its structural class, but specific Ames test data are not available. In acute toxicity studies, ALR2 inhibitors typically have high LD₅0 values (>2000 mg/kg in rodents). There is no evidence of organ-specific toxicity. Because ALR2 is not essential for normal physiology under normoglycemic conditions (ALR2 knockout mice are viable and healthy), selective ALR2 inhibition is expected to have a wide safety margin. However, caution should be exercised as with any new chemical entity. Standard laboratory safety practices (gloves, lab coat, safety glasses) should be followed. The compound should be stored at -20degC in a desiccator, protected from light. ALR2-IN-7 is not a controlled substance and is for research use only.
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| References | |
| Additional Infomation |
ALR2-IN-7 is also known as Compound 5a and has the IUPAC name: 2-[(9H-fluoren-9-yl)carbonyl]-1,3-dioxo-2,3-dihydro-1H-isoindole-5-carboxylic acid or similar (exact structure may vary). The compound belongs to the class of fluorenyl-phthalimide hybrids. The synthesis was reported in Bioorganic Chemistry (2025) by Gundogdu S et al. ALR2-IN-7 is supplied as a white to off-white solid with purity >99% (by HPLC). The compound is stable at room temperature for short periods but should be stored long-term at -20degC. For solution preparation, dissolve in DMSO (17.5 mg/mL) to make a 62 mM stock. The compound is light-sensitive; protect from light. ALR2-IN-7 represents a promising lead for the development of novel therapies for diabetic complications and potentially for cancer treatment where ALR2 is involved in drug resistance and metastasis. As a research tool, it is valuable for studying the polyol pathway and its role in metabolic diseases. For research use only, not for human therapeutic application.
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| Molecular Formula |
C17H15NO3
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|---|---|
| Molecular Weight |
281.31
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| Exact Mass |
281.105
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| CAS # |
59935-47-6
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| PubChem CID |
43147
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
2
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
21
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| Complexity |
409
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1C2=CC=CC=C2C3=C1C=C(C=C3)NC(=O)CCC(=O)O
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| InChi Key |
GTWRPNCMRGJTMD-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C17H15NO3/c19-16(7-8-17(20)21)18-13-5-6-15-12(10-13)9-11-3-1-2-4-14(11)15/h1-6,10H,7-9H2,(H,18,19)(H,20,21)
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| Chemical Name |
4-(9H-fluoren-2-ylamino)-4-oxobutanoic acid
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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) |
DMSO : ~17.5 mg/mL (~62.21 mM; with sonication)
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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 | 3.5548 mL | 17.7740 mL | 35.5480 mL | |
| 5 mM | 0.7110 mL | 3.5548 mL | 7.1096 mL | |
| 10 mM | 0.3555 mL | 1.7774 mL | 3.5548 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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