| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg | |||
| Other Sizes |
| Targets |
24(S)-Hydroxycholesterol targets the N-methyl-D-aspartate receptor (NMDAR) as a positive allosteric modulator. In mouse hippocampal neurons, it potentiates NMDA-induced currents with an EC50 of approximately 1.2 μM. The synthetic analog SGE-201 showed an EC50 of 0.11 μM. [3]
It also targets liver X receptors (LXRs) as an agonist, though it is not a critical activator of LXR target genes in vivo. [2] It is a substrate for CYP46A1 (the synthesizing enzyme) and is metabolized in the liver. [2] In retinal protection studies, exogenous 24S-HC at 1 μM prevented pressure-induced axonal injury and apoptotic RGC death. [4] |
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| ln Vitro |
24(S)-Hydroxycholesterol (10 μM) significantly potentiated NMDA receptor-mediated currents in cultured mouse hippocampal neurons, while cholesterol and other oxysterols (22(R)-HC, 20(S)-HC) were inactive at ≤10 μM. [3]
In human neuroblastoma SH-SY5Y cells, 50 μM 24S-HC induced cell death, which was suppressed by γ-tocopherol but not by γ-tocotrienol. 24S-HC also induced phosphorylation of CaMKII (Thr286) and RIPK1. α-Tocopherol (50 μM) significantly suppressed 24S-HC-induced CaMKII phosphorylation but not RIPK1 phosphorylation. [1] In HEK293 cells expressing recombinant NMDARs, 24S-HC (1 μM) potentiated NMDA responses, with synthetic analogues SGE-201 and SGE-301 showing stronger effects. The potentiation was not subunit-selective (GluN2A, 2B, 2C, 2D all showed significant potentiation). [3] In excised outside-out membrane patches from hippocampal neurons, SGE-201 (0.2 μM) robustly potentiated NMDA channel activity, increasing NPo values (channel number/open probability) without changing single-channel current, indicating direct allosteric modulation. [3] In ex vivo rat retinas, 75 mmHg hydrostatic pressure increased CYP46A1 expression and 24S-HC levels (measured by LC-MS/MS). Exogenous 1 μM 24S-HC prevented high pressure-induced axonal swelling and RGC apoptosis. Conversely, the CYP46A1 inhibitor voriconazole (10 μM) caused severe excitotoxic retinal damage at normobaric pressure, which was prevented by co-administration of 30 μM 24S-HC. [4] |
| ln Vivo |
In a transgenic mouse model overexpressing human CYP46A1 (C46-HA mice), brain 24S-HC levels were increased approximately twofold (from ~15 to ~30 ng/mg tissue), plasma levels increased fivefold, and fecal excretion increased four- to sevenfold. Despite elevated 24S-HC, no significant activation of LXR target genes (e.g., Cyp7a1, Abca1) was observed in brain or liver. Brain cholesterol precursor levels (lathosterol, etc.) increased, indicating compensatory upregulation of cholesterol synthesis. [2]
In rats, subchronic PCP treatment (5 mg/kg twice daily for 7 days) caused social interaction deficits and novel object recognition impairment. Acute administration of the synthetic 24S-HC derivative SGE-301 (1, 3, 10 mg/kg, i.p.) significantly reversed these cognitive and social deficits. SGE-301 (3 and 10 mg/kg) increased social interaction time from 32 s (vehicle) to 57 s and 48 s respectively. For novel object recognition, SGE-301 at 1 and 3 mg/kg significantly improved discrimination ratios. [3] In a mouse Y-maze spontaneous alternation test, MK-801 (0.25 mg/kg, i.p.) impaired performance. SGE-201 (3, 10, 30 mg/kg, i.p.) dose-dependently reversed MK-801-induced deficits. [3] |
| Cell Assay |
SH-SY5Y cell viability assay (WST-8) : Cells were treated with 50 μM 24S-HC for 24 h with or without vitamin E analogs. Cell viability was measured using WST-8 reagent. 24S-HC significantly decreased viability, and γ-tocopherol (up to 50 μM) dose-dependently suppressed this effect. [1]
Lactate dehydrogenase (LDH) assay: After 24S-HC treatment, LDH release was measured to assess cell death. CaMKII inhibitors mM3 (20 μg/mL) significantly inhibited 24S-HC-induced cell death, while KN62 (10 μg/mL) did not. [1] Western blotting: Whole cell lysates were immunoblotted with anti-phospho-CaMKII (Thr286), anti-CaMKII, anti-RIPK1, and anti-β-actin. 24S-HC induced CaMKII phosphorylation (53 kDa band), which was reduced by mM3 and by the ACAT1 inhibitor K-604 (5 μM). RIPK1 knockdown by siRNA reduced total CaMKII levels and its phosphorylation. [1] Nile red staining for lipid droplet-like structures: SH-SY5Y cells treated with 50 μM 24S-HC for 6 h showed increased lipid droplet-like structures, which were not inhibited by γ-tocopherol (50 μM) or γ-tocotrienol (10 μM). [1] Whole-cell patch-clamp in hippocampal neurons: Cultured neurons were clamped at -70 mV in Mg²⁺-free saline with 0.5 μM glycine. 24S-HC (0.1-30 μM) was preapplied for 40-90 s before 10 μM NMDA application. Currents were potentiated with an EC50 of 1.2 μM. [3] Excised outside-out patch recording: Membrane patches were excised from hippocampal neurons. NMDA (300 μM) was applied, and SGE-201 (0.2 μM) added. NPo analysis showed increased channel open probability. [3] HEK293 recombinant NMDAR assay: HEK293 cells expressing human GluN1 and GluN2 subunits were tested with 30 μM NMDA + 5 μM glycine. 24S-HC (1 μM) significantly potentiated currents. [3] |
| Animal Protocol |
Transgenic mouse model (C46-HA) : Human CYP46A1 cDNA under chicken β-actin promoter was injected into fertilized mouse eggs. Mice were backcrossed with C57Bl/6Ncrl for 7 generations. Animals were killed by CO₂ inhalation; brain, liver, plasma collected. 24S-HC levels measured by GC-MS. [2]
Rat subchronic PCP model: Male Long-Evans rats received PCP (5 mg/kg, i.p.) twice daily for 7 days, followed by a 7-day washout. On day 14, SGE-301 (3, 10, 30 mg/kg in 30% Captisol + 0.01% Tween 80) was administered i.p. 60 min before social interaction test. For novel object recognition, SGE-301 (1, 3, 10 mg/kg) was given 60 min before training. Positive control: risperidone 0.2 mg/kg i.p. [3] Mouse Y-maze test: Male Swiss CD-1 mice received SGE-201 (3, 10, 30 mg/kg i.p. in 25% hydroxypropyl-β-cyclodextrin in PBS) 1 hour before testing. MK-801 (0.25 mg/kg i.p.) was given 30 min before testing. Spontaneous alternation was recorded for 10 min. [3] Rat ex vivo retinal pressure model: Eyecups from 28-32 day old rats were placed in aCSF in a closed pressure chamber at 10, 35, or 75 mmHg for 24 h at 30°C. 24S-HC (1, 10, 30 μM) or voriconazole (1, 10 μM) was added to the aCSF. [4] |
| ADME/Pharmacokinetics |
In C46-HA transgenic mice overexpressing human CYP46A1, brain 24S-HC content was approximately 30 ng/mg tissue (twice control), plasma levels ~150 ng/mL (fivefold control), and fecal excretion of free 24S-HC was 4-7 times higher than wild-type (approximately 1.5-2.5 μg/24h vs. 0.3-0.4 μg/24h). [2]
In rats, following intraperitoneal administration of SGE-301 (10 mg/kg), brain concentrations reached ~200 ng/g and plasma concentrations ~300 ng/mL (determined by LC-MS/MS). [3] In human brain homogenates, endogenous 24S-HC levels are reported to be in the tens of micromolar concentration range (approximately 25 μM). [4] |
| Toxicity/Toxicokinetics |
In SH-SY5Y cells, 50 μM 24S-HC induced cell death via necroptosis-like pathways (RIPK1-dependent, caspase-independent). This was not mediated by ROS generation. [1]
In the rat ex vivo retina, the CYP46A1 inhibitor voriconazole (10 μM) caused severe excitotoxic retinal damage characterized by edematous changes in the inner plexiform layer and bull's eye formation in the inner nuclear layer, which was prevented by co-administration of 30 μM 24S-HC. [4] No in vivo systemic toxicity data (e.g., LD50) for 24S-HC itself are reported in these studies. The synthetic derivatives SGE-201 and SGE-301 were well-tolerated at the doses used (up to 30 mg/kg i.p. in mice/rats). [3] |
| References |
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| Additional Infomation |
(24S)-24-hydroxycholesterol is a 24-hydroxycholesterol with an S configuration at position 24. It is a major metabolic breakdown product of cholesterol in the brain. It is a metabolite in mice, as well as a biomarker and a human serum metabolite. Reports and data exist regarding 24-hydroxycholesterol in humans.
Biosynthesis: 24S-HC is synthesized from cholesterol by CYP46A1, an enzyme expressed predominantly in neurons of the CNS and retina. [2][4] Mechanism of NMDAR modulation: 24S-HC acts as a positive allosteric modulator at NMDARs, stabilizing the inactive conformation of the receptor? (It potentiates currents but also shows slow onset and reversibility). It binds to a site distinct from pregnenolone sulfate, as occlusion studies showed 24S-HC did not occlude PREGS potentiation. [3] Role in retinal protection: Elevated hydrostatic pressure (75 mmHg) increases CYP46A1 expression and 24S-HC synthesis in the retina. Exogenous 24S-HC (1 μM) protects against pressure-induced axonal swelling and RGC apoptosis. Voriconazole, a CYP46A1 inhibitor, is retinotoxic at normobaric pressure, and this toxicity is prevented by 24S-HC. [4] Neuroprotection vs. neurotoxicity: 24S-HC has dual effects: at high concentrations (≥50 μM) it induces necroptosis-like cell death, while at low micromolar concentrations (1-10 μM) it exerts neuroprotective effects against excitotoxicity and pressure-induced damage. [1][3][4] Synthetic derivatives: SGE-201 and SGE-301 are synthetic analogs with improved potency (EC50 ~0.11 μM for SGE-201) and in vivo efficacy. SGE-301 has a 3α-methyl group designed to improve bioavailability. [3] |
| Molecular Formula |
C27H46O2
|
|---|---|
| Molecular Weight |
402.652948856354
|
| Exact Mass |
402.349
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| Elemental Analysis |
C, 80.54; H, 11.52; O, 7.95
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| CAS # |
474-73-7
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| Related CAS # |
24-Hydroxycholesterol;30271-38-6
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| PubChem CID |
121948
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| Appearance |
White to off-white solid powder
|
| Density |
1.0±0.1 g/cm3
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| Boiling Point |
513.1±23.0 °C at 760 mmHg
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| Melting Point |
174-176°C (lit.)
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| Flash Point |
213.5±17.2 °C
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| Vapour Pressure |
0.0±3.0 mmHg at 25°C
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| Index of Refraction |
1.536
|
| LogP |
7.66
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| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
2
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
29
|
| Complexity |
624
|
| Defined Atom Stereocenter Count |
9
|
| SMILES |
O[C@H]1CC[C@@]2(C)C(C1)=CC[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)CC[C@@H](C(C)C)O)CC[C@H]21
|
| InChi Key |
IOWMKBFJCNLRTC-XWXSNNQWSA-N
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| InChi Code |
InChI=1S/C27H46O2/c1-17(2)25(29)11-6-18(3)22-9-10-23-21-8-7-19-16-20(28)12-14-26(19,4)24(21)13-15-27(22,23)5/h7,17-18,20-25,28-29H,6,8-16H2,1-5H3/t18-,20+,21+,22-,23+,24+,25+,26+,27-/m1/s1
|
| Chemical Name |
(3S,8S,9S,10R,13R,14S,17R)-17-[(2R,5S)-5-hydroxy-6-methylheptan-2-yl]-10,13-dimethyl-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-ol
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| Synonyms |
24(S)-hydroxycholesterol; 474-73-7; 24S-hydroxycholesterol; 24S-OHC; 47IMW63S3F; 24S-OH-cholesterol; 24S-OHC; 24S-HC; Cerebrosterol; 24(S)-Saringosterol; 24S-Saringosterol; 24(S) Saringosterol
|
| 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) |
Ethanol : ~20 mg/mL (~49.67 mM)
DMSO : ~0.1 mg/mL (~0.25 mM) |
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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 | 2.4835 mL | 12.4177 mL | 24.8355 mL | |
| 5 mM | 0.4967 mL | 2.4835 mL | 4.9671 mL | |
| 10 mM | 0.2484 mL | 1.2418 mL | 2.4835 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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