| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
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| Targets |
somatostatin sst4 receptor
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|---|---|
| ln Vitro |
The discovery of novel non-peptide compounds with a high affinity for the peptide hormone somatostatin (SST) receptor is described. The compounds were tested for affinity at five human SST receptor subtypes individually expressed in mammalian cells. The compound NNC 26-9100 showed a Ki of 6 nM at SST4 and more than 100 fold selectivity for SST4 over SST1, SST2, SST3, or SST5. Competition binding studies and Scatchard analysis of the interaction by NNC 26-9100 with SST showed specificity at SST4. Furthermore, NNC 26-9100 was highly selective for SST4 over a variety of other G protein-coupled receptors, having affinities for M1 muscarinic acetylcholin and D3 dopamine receptors of around 500 and 1000 nM, respectively. Finally, NNC 26-9100 was found to fully inhibit forskolin-induced accumulation of adenosine 3‘,5‘-cyclic monophosphate in baby hamster kidney cells, expressing the human SST4 receptor with an EC50 of 2 nM.
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| ln Vivo |
In this study, researchers evaluated the effect of selective SSTR4 agonist NNC 26-9100 on the changes in learning and soluble Aβ42 oligomer brain content with and without co-administration of the M13-metalloproteinase family enzyme-inhibitor phosphoramidon, using the senescence-accelerated mouse prone-8 (SAMP8) model. NNC 26-9100 treatment (0.2 µg i.c.v. in 2 µL) improved learning, which was blocked by phosphoramidon (1 and 10mM, respectively). NNC 26-9100 decreased total soluble Aβ42, an effect which was blocked by phosphoramidon (10mM). Extracellular, intracellular, and membrane fractions were then isolated from cortical tissue and assessed for soluble oligomer alterations. NNC 26-9100 decreased the Aβ42 trimeric (12 kDa) form within the extracellular and intracellular fractions, and produced a band-split effect of the Aβ42 hexameric (25 kDa) form within the extracellular fraction. These effects were also blocked by phosphoramdon (1 and 10mM, respectively). Subsequent evaluation of NNC 26-9100 in APPswe Tg2576 transgenic mice showed a similar learning improvement and corresponding reduction in soluble Aβ42 oligomers within extracellular, intracellular, and membrane fractions. These data support the hypothesis that NNC 26-9100 reduces soluble Aβ42 oligomers and enhances learning through a phosphoramidon-sensitive metalloproteinase-dependent mechanism[2].
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| Enzyme Assay |
Binding Assay. [1]
Cells expressing individual SST receptor subtypes were resuspended in buffer (50 mM Tris−HCl (pH 7.4), 1 mM EGTA, 5 mM MgCl2), and homogenized. Membranes were washed twice in buffer by homogenization and centrifugation. The final membrane pellets were resuspended at a protein concentration of 125 μg/mL in buffer. Binding assays using 75 pM 125I-Tyr11-SRIF (Amersham, IM-161) were done in duplicates in minisorb polypropylene tubes in a volume of 250 μl. The assays were incubated at 30-37 °C for 30-90 min depending on the receptor subtype. Binding was terminated by filtration through Whatman GF/B glass fiber filters presoaked for 4 h in 0.5% poly(ethylenimine) and 0.1% BSA. The filters were washed three times with 5 mL of ice-cold 0.9% saline and counted in a Packard Cobra II gamma counter. Assays for other G protein-coupled receptors were performed by a CRO company with the radioligands mentioned in Table 2 using plasma membrane preparations from rabbit adrenal gland (ATII), guinea pig ileum (B2), rat pancreas (CCKA), mouse brain (CCKB), human recombinant (D3), A10 cells (ETA), rat cerebellum (ETB), rat brain (galanin), rat brain cortex (M1), rat heart (M2), guinea pig submaxillary gland (NK1), rabbit kidney medulla (Y2), rat brain cortex (5-HT1A), rabbit ileum (5-HT3), and guinea pig lung (VIP). FunctionalAssay. [1] Cells expressing human SST receptors were seeded in 24-well tissue culture multidishes at 200 000 cells/well and grown for 16-20 h. The medium was removed, and fresh DMEM medium, supplemented with (1) 3-isobutyl-1-methylxanthine (IBMX), (2) forskolin or medium, and (3) medium, SRIF, SST analogue, or compound, was added. The plates were incubated for 15-30 min at 37 °C, the reaction medium was removed, and the cells were lysed with 0.1 M sodium hydroxide. Following neutralization with 0.1 M hydrochloric acid, an aliquot was removed for c-AMP determination using Amersham SPA RIA (RPA 538). |
| Cell Assay |
BiologicalAssay.[1]
Cell Lines Expressing SST Receptor Subtypes. BHK cells (tk-ts13, ATCC CRL No. 1632) and HEK 293 cells (ATCC CRL No. 1573) were grown to 20−40% confluency in a tissue culture dish in DMEM containing 1% penicillin/streptomycin, 10% fetal bovine serum, and 1% Glutamax. Prior to transfection, the cells were washed twice with calcium-free PBS, after which 20 mL of serum-free DMEM was added to the cells. Transfection was carried out as described previously (product description: lipofectamin). Briefly, 10 μg of c-DNA encoding a SST-receptor subtype inserted into the mammalian expression vector pcDNA3 was diluted in 300 μL of sterile water. Lipofectamin (30 μg) was diluted in 300 μL of sterile water. The c-DNA and lipofectamin solutions were mixed and left at room temperature for 15 min. The lipofectamin/c-DNA mixture was added dropwise to the cells (HEK 293 cells for SST2, BHK for the other receptor subtypes) while the plates were gently swirled. The cells were then incubated for 16-24 h, after which the medium was replaced with standard medium containing 1 mg/mL Geneticin (G-418 sulfate). Resistant colonies appearing after 1-2 weeks were isolated and propagated for further characterization. |
| Animal Protocol |
Animals[2]
Twelve-month old male SAMP8 or APPswe Tg2576 (model 002789) mice were used for these evaluations. Mice were housed in rooms with a 12 h light/dark cycle (20–22°C) with water and food available ad libitum. All experiments were conducted in accordance with the institutional approval of the animal use subcommittee, which subscribes to the NIH Guide for Care and Use of Laboratory Animals. Dosing[2] NNC 26-9100 treatment was performed via intracerebroventricular (i.c.v.) injection. The i.c.v. dose of NNC 26-9100 (0.2 μg) previously identified to enhance learning in 12-month old SAMP8 mice (Sandoval et al., 2012) was tested against the M13 family inhibitor phosphoramidon (1 or 10 mM) (American Peptide Co, Sunnyvale, CA) via co-administration, and evaluated against respective controls. Previous studies have shown phosphoramidon (i.c.v.) to elevate soluble Aβ levels within 2 h (Eckman et al., 2006). Briefly, 24 h prior to T-maze testing, the mice were anesthetized with isoflurane, placed in a stereotaxic instrument. The scalp was opened and a unilateral hole was drilled 0.5 mm anterior and 1.0 mm to the right of the bregma. A single i.c.v. (2 μL) injection of NNC 26-9100 (0.2 μg) with or without phosphoramidon (1 or 10 mM), or vehicle control (20% ethanol/saline), was conducted with a 30 g blunt needle connected to a Hamilton syringe, to a depth of 2 mm. Syringe was kept in place via side-arm attachment. Injections were conducted in a gradual manner over ~1 min and syringe was kept in place for 3 s following to assure no back-flow occurred. The hole was filled in with bone-wax and scalp sealed via Vetbond. Subsequent to behavioral evaluations and immediately prior to respective tissue assessments brains were cut along the injection site to assure accuracy. |
| References |
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| Additional Infomation |
1-[3-[(5-bromo-2-pyridinyl)-[(3,4-dichlorophenyl)methyl]amino]propyl]-3-[3-(1H-imidazol-5-yl)propyl]thiourea is an aminopyridine.
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| Molecular Formula |
C22H25BRCL2N6O
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|---|---|
| Molecular Weight |
540.29
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| Exact Mass |
554.042
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| CAS # |
199522-35-5
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| PubChem CID |
9893924
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| Appearance |
Typically exists as solid at room temperature
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| Density |
1.482g/cm3
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| Boiling Point |
731.167ºC at 760 mmHg
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| Flash Point |
395.996ºC
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| Vapour Pressure |
0mmHg at 25°C
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| Index of Refraction |
1.668
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| LogP |
6.149
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
11
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| Heavy Atom Count |
32
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| Complexity |
563
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C=NN
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| InChi Key |
UREJDUPKGMFJRU-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C22H25BrCl2N6S/c23-17-5-7-21(29-12-17)31(14-16-4-6-19(24)20(25)11-16)10-2-9-28-22(32)27-8-1-3-18-13-26-15-30-18/h4-7,11-13,15H,1-3,8-10,14H2,(H,26,30)(H2,27,28,32)
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| Chemical Name |
1-[3-[(5-bromopyridin-2-yl)-[(3,4-dichlorophenyl)methyl]amino]propyl]-3-[3-(1H-imidazol-5-yl)propyl]thiourea
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| Synonyms |
NNC-269100; NNC 269100; 199522-35-5; ML156; CHEMBL103769; 9X9D329ZX2; 1-[3-[(5-bromo-2-pyridinyl)-[(3,4-dichlorophenyl)methyl]amino]propyl]-3-[3-(1H-imidazol-5-yl)propyl]thiourea; NNC269100
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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 | 1.8509 mL | 9.2543 mL | 18.5086 mL | |
| 5 mM | 0.3702 mL | 1.8509 mL | 3.7017 mL | |
| 10 mM | 0.1851 mL | 0.9254 mL | 1.8509 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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