Compounds, compositions and methods of inhibiting alpha-synuclein toxicity
A technology for synuclein and toxicity, which can be used in drug combinations, active ingredients of heterocyclic compounds, pharmaceutical formulations, etc., and can solve problems such as the ambiguous relationship between synuclein pathology and neurodegenerative diseases
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[0304] C. Preparation of compounds
[0305] The compounds described for use in the compositions and methods provided herein are available from commercial sources (eg, Aldrich Chemical Co., Milwaukee, WI), can be prepared by methods well known to those skilled in the art or as shown herein. One skilled in the art should be able to prepare all of the compounds used herein by routine modification of the above methods using appropriate starting materials.
[0306] Some of the compounds provided herein can be prepared by the synthetic routes shown below. Briefly, aryl or heteroaryl amines are converted to 1 using the corresponding nitrile. Compound 1 can also be synthesized in other ways, including starting from aryl halides or heteroaryl halides, using the corresponding guanidinium salts. Treatment of compound 1 with an acid halide or anhydride affords the corresponding acylated compound 2, which can be converted to the corresponding amide 3 by reaction with ammonia. Compound 1...
Embodiment 1
[0406] α-Synuclein (aS) Screening
[0407] yeast strain
[0408] Parent W303: MAT a / α ade2-1 / ade2-1 his3-11, 15 / his3-11, 15 leu2-3, 112 / leu2-3, 112
[0409] trp1-1 / trp1-1 ura3-1 / ura3-1 can1-100 / can1-100
[0410] Phenotype: Requires adenine, histidine, leucine, tryptophan, and uracil for growth. Resistant to canavanine.
[0411] Fx-109: MAT a / α ade2-1 / ade2-1 his3-11, 15 / his3-11, 15leu2-3, 112 / leu2-3, 112
[0412] trp1-1 / trp1-1 GALp-aS-GFP::TRP1 / GALp-aS-GFP::TRP1 ura3-1 / ura3-1
[0413] GALp-aS-GFP::URA3 / GALp-aS-GFP::URA3 can1-100 / can1-100pdr1::KanMX / pdr1::KanMX erg6::KanMX / erg6::KanMX
[0414] Phenotype: Inability to grow on galactose due to expression of aS. Requires histidine, leucine, and adenine for growth. Resistant to canavanine and kanamycin. Highly sensitive to drugs.
[0415] Media and Reagents
[0416] Depending on the genotype of the strain to be tested, select appropriate supplements for the synthetic media. Strains containing an integrating construct (eg ...
Embodiment 2
[0435] Some compounds according to formulas I-IX can be prepared as follows.
[0436] Preparation of guanidine
[0437] method 1
[0438]
[0439] The mixture of guanidine carbonate (22g, 120mmol) and 2-chlorobenzoxazole (7ml, 60mmol) in N, N-diisopropylethylamine (45ml, 259mmol) and DMF (100ml) solution was at 60°C Heat overnight. After evaporation it was treated with water (100ml). The ppts were collected, washed with ether and dried. Yield of desired intermediate product: 9.3 g (88%), beige solid. Purity: 95%.
[0440] Method 2
[0441]
[0442] Mix 2-aminobenzothiazole 1 (1eq) and 2-methyl-2-thiopseudourea sulfate (2-methyl-2-thiopseudourea sulfate) (1eq) in a vial, in an oil above the melting point of the mixture Bath (140-220 ° C) was heated with stirring for 2h. After cooling the mixture was triturated well with hot MeOH, filtered and washed with MeOH. The resulting gray solid product (presumably in the mixed sulfate form with the S-methyl-2-thiopseudoure...
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