Nitrogen-containing heterocyclic compound for treating brain injury and application thereof
A compound and brain injury technology, applied in the direction of organic chemistry, drug combination, medical preparations containing active ingredients, etc., to achieve the effect of prevention and treatment of brain injury, good therapeutic activity, and broaden the scope
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Embodiment 1
[0025] Embodiment 1: the synthesis of compound 6
[0026] (1) Dissolve compound 1 (166g, 1mol) in dry dichloromethane (500mL) in a reaction flask, add triethylamine (10mol), and add compound 2 (345g, 1.2mol) di Chloromethane (600mL) solution was added dropwise to room temperature and reacted until the reaction was detected by TLC. The reaction solution was suction filtered, the filtrate was concentrated, and silica gel column chromatography (ethyl acetate / petroleum ether=3:1) gave compound 3. The yield was 89%, HPLC purity 98%;
[0027] (2) Add compound 3 (206g, 0.5mol), palladium catalyst Pd(t-Bu 3 P) 2 (6.4g, 0.0125mol) and hexamethyltin (197g, 0.6mol) and 500mL of toluene, heated to reflux and reacted until TLC detected that the reaction of raw materials was complete, quenched with water, extracted and separated, retained the organic phase, and used fluorinated organic phase Washed twice with potassium-saturated ammonia solution, dried over anhydrous magnesium sulfate, f...
Embodiment 2
[0031] Embodiment 2: the effect of the compound shown in formula 6 in treating brain injury
[0032] 1. Animals and materials
[0033] Compound 6, self-made, HPLC purity 99.0%; DW-2000 brain stereotaxic apparatus (Chengdu Taimeng Software Co., Ltd.); SD male rats, SPF grade, 180-220g, 6-8 weeks old (Zhejiang Experimental Animal Center) ; Neonatal mice (Zhejiang Experimental Animal Center); IL-6, IL-1, TNF-α kit (USA eBio-science company); neuron β-tubulin III antibody (USA Co-vanc company); Alexa Fluor 488 goat anti mouse IgG dye and Hoeschst dye (Invitrogen, USA); FBS (Gibco, USA).
[0034] 2. Experimental method
[0035] 2.1 Transwell experiment
[0036] Newborn mice within 1 day were taken out, and the cerebral cortex was separated in an ice bath, digested with 1% trypsin into a single cell suspension, microglial cells were cultured with 10% FBS+DMEM / F12, and neurons were cultured with 2% FBS+DMEM / F12 culture, change the medium every 3 days, purify after 2 weeks, divid...
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