CYCLIC NUCLEIC ACID MOLECULE HAVING GENE EXPRESSION CONTROL FUNCTION
FIG3 is an RP-HPLC chart of PA-0002-DSP
120 µmol/L PA-0002 (350 µL), 10 mg/mL DSP-ONSu/DMF solution (65 µL), isopropanol (250 µL) and 1% triethylamine aqueous solution (35 µL) were mixed and stirred at 40°C for 4 hr. The reaction mixture was purified by HPLC (column: Develosil C8-UG-5, 5 µm, 10×50 mm; flow rate: 4.7 mL/min; detection: UV260 nm; column oven: 40°C; Buffer A: 50 mmol/L TEAA (pH 7.0), 5% CH3CN; Buffer and the peak of the desired product was collected. The collected fraction was subjected to ethanol precipitation and the resulting precipitate was dissolved in distilled water for injection (Otsuka Pharmaceutical, Co., Ltd.). Absorbance at UV260 nm was measured and the yield was calculated. PA-0002-DSP (0.25 mg) with purity 89.32% was obtained. Mass spectrometry: 16881.90 (Calculated: 16882.50). The HPLC chart after purification is shown in FIG3.
flowchart
67
with purity
3
Example
10
- reaction mixture was added
20
-
['13. The method for inhibiting expression according to claim 9 or 10 wherein the nucleic acid molecule is administered to a non-human animal.']