SGE516_Others_Signaling Pathways_Products

SGE-516, a synthetic neuroactive steroid, is a potent positive allosteric modulator of both synaptic and extrasynaptic GABAA receptors. Neuroactive steroids (NASs) have been shown to impact central nervous system (CNS) function through positive allosteric modulation of the GABA(A) receptor (GABA(A)-R). SGE-516 treatment protected against hyperthermia-induced seizures, reduced spontaneous seizure frequency and prolonged survival in the Scn1a +/- mice. This provides the first evidence of SGE-516 activity in a mouse model of Dravet syndrome, and supports further investigation of neuroactive steroids as potential anticonvulsant compounds for refractory epilepsies. As a balanced synaptic/extrasynaptic GABA(A) receptor modulator, and a selective extrasynaptic GABA(A) receptor modulator, SGE-516 possesses excellent druglike properties, making them advanced leads for oral delivery of GABA(A) receptor modulators.

Physicochemical Properties

Related CAS #

N/A

Appearance

Typically exists as solid at room temperature

Chemical Name

1-((3R,5R,8R,9R,10S,13S,14S,17S)-3-hydroxy-3,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)-2-(2H-1,2,3-triazol-2-yl)ethan-1-one

Synonyms

SGE-516; SGE516; SGE 516

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)

Biological Activity

ln Vitro

In vitro activity: SGE-516, a synthetic neuroactive steroid, is a potent positive allosteric modulator of both synaptic and extrasynaptic GABAA receptors. Neuroactive steroids (NASs) have been shown to impact central nervous system (CNS) function through positive allosteric modulation of the GABA(A) receptor (GABA(A)-R). SGE-516 treatment protected against hyperthermia-induced seizures, reduced spontaneous seizure frequency and prolonged survival in the Scn1a +/- mice. This provides the first evidence of SGE-516 activity in a mouse model of Dravet syndrome, and supports further investigation of neuroactive steroids as potential anticonvulsant compounds for refractory epilepsies. As a balanced synaptic/extrasynaptic GABA(A) receptor modulator, and a selective extrasynaptic GABA(A) receptor modulator, SGE-516 possesses excellent druglike properties, making them advanced leads for oral delivery of GABA(A) receptor modulators.

Kinase Assay:

Cell Assay: Hyperthermia-induced seizure assays were performed on Scn1a +/− mice at P19-20. Core body temperature was monitored with a RET-3 rectal temperature probe (Physitemp Instruments, Inc, New Jersey, USA) and controlled by a heat lamp connected to a rodent temperature regulator (TCAT-2DF, Physitemp) reconfigured with a Partlow 1160 + controller (West Control Solutions, Brighton, UK). SGE-516 was administered by IP injection and mice were returned to their home cage for 40 minutes. The temperature probe was inserted at 40 minutes post-injection and mice acclimated to probe and test environment for 5 minutes. At 45 minutes post-injection, body temperature was raised 0.5 °C every two minutes until the onset of the first clonic convulsion with loss of posture or until 42.5 °C was maintained for 5 minutes. Mice that did not experience a GTCS during the 5-minute hold at 42.5 °C were considered seizure-free. Threshold temperatures were compared using the time-to-event analysis with P values determined with LogRank Mantel-Cox test. P values

ln Vivo

Scn1a tm1Kea mice, with deletion of the first coding exon, were generated by homologous recombination in TL1 ES cells (129S6/SvEvTac) and genotyped as previously described. The Scn1a +/− line is maintained by continuous backcrossing to 129S6/SvEvTac (129) (Taconic Biosciences, Hudson, NY, USA). Mice for experiments were generated by crossing 129.Scn1a +/− mice with C57BL/6 J (B6) (#000664, Jackson Laboratory, Bar Harbor, ME, USA), resulting in [129 x B6]F1.Scn1a +/− offspring, referred to herein as Scn1a +/− mice. Mice were maintained in a Specific Pathogen Free (SPF) barrier facility with a 14-hour light/10-hour dark cycle and access to food and water ad libitum. Both female and male Scn1a +/− mice were utilized for all experiments. There were no significant differences between sexes on any measurements, so groups were collapsed across sex. All animal care and experimental procedures were approved by the Northwestern University Animal Care and Use Committee in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals. The principles outlined in the ARRIVE (Animal Research: Reporting of in vivo Experiments) guideline and Basel declaration (including the 3 R concept) were considered when planning experiments.

Animal Protocol

SGE-516 was solubilized in 2% 2-Hydroxypropyl-β-cyclodextrin (HPBCD) by sonication until a clear solution was obtained and was administered by intraperitoneal (IP) injection; 8, 40, 120 or 267 mg/kg/day

Scn1a tm1Kea mice

References

J Med Chem.2015 Apr 23;58(8):3500-11;Sci Rep.2017 Nov 10;7(1):15327.

Solubility Data

Solubility (In Vitro)

DMSO: 10 mM

Water:< 1mg/mL

Ethanol:< 1mg/mL

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
(e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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).
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)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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).
Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

(Please use freshly prepared in vivo formulations for optimal results.)

PeptideDB

SGE516

CAS No.:

Physicochemical Properties

Biological Activity

Solubility Data