![tert-Butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate](/upload/0201/V105495.png)
| Size | Price | Stock | Qty |
|---|---|---|---|
| 5g |
|
||
| 10g |
|
||
| 25g | |||
| Other Sizes |
| Targets |
tert-Butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate has no direct biological target; it is a PROTAC linker. As a chemical building block, it serves as a rigid spacer that connects an E3 ubiquitin ligase ligand (e.g., CRBN or VHL) to a target protein ligand in PROTAC molecules. The spirocyclic core provides conformational rigidity, which can influence the three-dimensional orientation and optimal distance between the two ligands, thereby affecting ternary complex formation and degradation efficiency. The Boc group protects the piperazine nitrogen during synthesis and can be removed under acidic conditions (e.g., TFA in DCM) to reveal a secondary amine for further conjugation. The free amine can be used for amide bond formation with carboxylic acid-containing ligands. The rigid spiro linker has been used in PROTAC ER Degrader-12 and PROTAC AR Degrader-9.
|
|---|---|
| ln Vitro |
The compound itself has no direct in vitro biological activity. It is a chemical building block for PROTAC synthesis. Its activity is assessed after conjugation to an E3 ligase ligand and a target protein ligand. The resulting PROTAC is then evaluated for its ability to induce target protein degradation in cells. The rigid spiro linker may improve PROTAC potency by pre-organizing the geometry for optimal ternary complex formation. For PROTAC evaluation, cells expressing the target protein are seeded in 6-well plates and treated with the PROTAC (0.1-1000 nM) for 4-24 h. Target protein degradation is assessed by Western blotting. DC₅0 (half-maximal degradation concentration) and Dmax (maximal degradation) are calculated.
|
| ln Vivo |
The compound is not a drug; it is a PROTAC linker. PROTACs synthesized using this rigid spiro linker can have potent in vivo activity. In xenograft mouse models, PROTACs are administered intraperitoneally or intravenously (10-100 mg/kg) once daily for 2-4 weeks. They induce sustained degradation of target proteins in tumors (>80% reduction) and inhibit tumor growth. The spirocyclic linker contributes to the overall stability, rigidity, and pharmacokinetic properties of the PROTAC. The Boc group is removed during PROTAC synthesis and is not present in the final molecule.
|
| Enzyme Assay |
Not applicable. tert-Butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate is a PROTAC linker, not a standard enzyme inhibitor. Its purity (>98%) is assessed by HPLC. The structure is confirmed by ¹H NMR (Boc group delta 1.4-1.5 ppm, spirocyclic protons delta 1.5-1.8 ppm, N-CH2 delta 2.4-3.5 ppm) and mass spectrometry (ESI-MS, expected [M+H]+ m/z 227). Physicochemical properties: MW 226.32, LogP 1.23, density 1.1 g/cm3, boiling point 319.4degC, flash point 147.0degC, white to off-white solid powder, soluble in DMSO. Storage: powder at -20degC for 3 years, 4degC for 2 years; in solvent at -80degC for 6 months, -20degC for 1 month. Protect from light.
|
| Cell Assay |
Not applicable. The compound is not used directly in cell-based assays; it is a building block for PROTAC synthesis. For PROTAC evaluation, cells expressing the target protein are seeded in 6-well plates (3×10⁵ cells/well) and treated with the PROTAC (0.1-1000 nM) for 4-24 h. Target protein degradation is assessed by Western blotting. DC₅0 is calculated from the dose-response curve. Cell viability is measured by MTT or CellTiter-Glo. The rigid spiro linker may affect PROTAC cellular permeability and degradation potency compared to flexible linkers.
|
| Animal Protocol |
In vivo efficacy of PROTACs incorporating tert-Butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate can be evaluated in xenograft mouse models. Female BALB/c nude mice (6-8 wk) are subcutaneously inoculated with 5×10⁶ cancer cells. When tumors reach ~100-150 mm3, mice are randomized into treatment groups (n=6-8/group). The PROTAC is formulated in 10% DMSO + 40% PEG300 + 5% Tween 80 + 45% saline and administered intraperitoneally at 10-100 mg/kg once daily or every other day for 2-4 weeks. Tumor volume is measured every 3 days with calipers, and tumors are excised for Western blot analysis of target protein levels.
|
| ADME/Pharmacokinetics |
No specific PK data is available for tert-Butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate alone. When incorporated into a PROTAC (MW 800-1200), the overall PK properties are determined by the full PROTAC molecule. PROTACs generally have moderate to low oral bioavailability (<20%), short plasma half-life (1-4 h), high plasma protein binding (>90%), and rapid clearance. The rigid spiro linker may improve metabolic stability and reduce off-target interactions compared to flexible aliphatic linkers. The Boc group is not present in the final PROTAC molecule.
|
| Toxicity/Toxicokinetics |
For tert-Butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate, hazard statements: H315 (Causes skin irritation), H319 (Causes serious eye irritation), H335 (May cause respiratory irritation). Signal word: Warning. Precautionary statements: P261 (Avoid breathing dust/fume/gas/mist/vapors/spray), P280 (Wear protective gloves/protective clothing/eye protection/face protection), P305+P351+P338 (IF IN EYES: Rinse cautiously with water for several minutes). Storage: powder at -20degC for 3 years, 4degC for 2 years; in solvent at -80degC for 6 months, -20degC for 1 month. Protect from light.
|
| References | |
| Additional Infomation |
tert-Butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (CAS# 236406-55-6) is a research-grade rigid spirocyclic PROTAC linker used in the synthesis of PROTAC ER Degrader-12 and PROTAC AR Degrader-9. It is not an FDA-approved drug. For research use only, not for diagnostic or therapeutic applications.
|
| Molecular Formula |
C12H22N2O2
|
|---|---|
| Molecular Weight |
226.32
|
| Exact Mass |
226.168
|
| CAS # |
236406-55-6
|
| PubChem CID |
23282935
|
| Appearance |
White to off-white solid powder
|
| Density |
1.1±0.1 g/cm3
|
| Boiling Point |
319.4±42.0 °C at 760 mmHg
|
| Flash Point |
147.0±27.9 °C
|
| Vapour Pressure |
0.0±0.7 mmHg at 25°C
|
| Index of Refraction |
1.518
|
| LogP |
1.23
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
3
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
16
|
| Complexity |
269
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
O(C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H])C(N1C([H])([H])C2(C([H])([H])C([H])([H])N([H])C([H])([H])C2([H])[H])C1([H])[H])=O
|
| InChi Key |
HWLNKJXLGQVMJH-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C12H22N2O2/c1-11(2,3)16-10(15)14-8-12(9-14)4-6-13-7-5-12/h13H,4-9H2,1-3H3
|
| Chemical Name |
tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate
|
| 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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| 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
|
|---|---|
| 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 | 4.4185 mL | 22.0926 mL | 44.1852 mL | |
| 5 mM | 0.8837 mL | 4.4185 mL | 8.8370 mL | |
| 10 mM | 0.4419 mL | 2.2093 mL | 4.4185 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
