Apoptosis PET imaging agents
An imaging agent and covalent linking technology, which is applied in the direction of in vivo radioactive preparations, X-ray contrast agent preparation, pharmaceutical formulations, etc., to achieve the effect of easy image quantification
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Embodiment 1
[0189] Example 1: Synthesis of (Boc-aminooxy)acetyl-duramycin (precursor 1A).
[0190]
[0191]Duramycin (Sigma-Aldrich; 8.0 mg, 4.0 μmol), TFP (Boc-aminooxy)acetate (Invitrogen; 1.3 mg, 3.8 μmol) and DIPEA (2.1 μL, 12.5 μmol) were dissolved in NMP (1 mL )middle. The reaction mixture was shaken for 30 minutes. The mixture was then diluted with water / 0.1% TFA (6 mL), and the product was purified using preparative HPLC.
[0192] By preparative HPLC (Beckman System Gold chromatography system, using the following conditions: solvent A = H 2 O / 0.1% TFA with solvent B = ACN / 0.1% TFA, Gradient: 20-50% B in 40 minutes; Flow rate: 10 mL / min; Column: Phenomenex Luna 5 μm C18 (2) 250 x 21.2 mm; Detection: UV 214 nm) purification afforded 3.8 mg of pure precursor 1A (yield 44%). Purified material was analyzed by analytical LC-MS (gradient: 20-70% B in 5 minutes, t R : 1.93 minutes, measured value m / z : 1093.7, expected MH 2 2+ : 1093.5) analysis.
[0193] Separation of...
Embodiment 2
[0195] Example 2: Synthesis of (Eei-aminooxy)acetyl-duramycin (precursor 1B).
[0196]
[0197] Duramycin (Sigma-Aldrich; 50 mg, 25 μmol), NHS (Eei-aminooxy)acetate (Iris Biotech., 5.1 mg, 20 μmol) and DIPEA (17 μL, 100 μmol) were dissolved in NMP ( 1 mL). The reaction mixture was shaken for 45 minutes. The mixture was then diluted with water / 0.1% acetic acid (8 mL), and the product was purified using preparative HPLC.
[0198] Purification by preparative HPLC (as for Example 1 with a gradient of 14-45% B over 40 minutes, where A=water / 0.1% acetic acid and B=ACN) afforded 14 mg of pure precursor 1B (yield 26% ). The purified material was analyzed by LC-MS (gradient: 20-50% B in 5 minutes, t R : 2.5 and 2.7 minutes, measured values m / z : 1078.8, expected MH 2 2+ : 1078.5) analysis.
[0199] Chromatographic resolution of (Eei-aminooxy)acetyl-duramycin regioisomers was achieved on analytical HPLC using 0.1% TFA. However, the Eei protecting group is unstable in...
Embodiment 3
[0200] Example 3: Synthesis of Aminooxyacetyl-Duramycin (Precursor 2).
[0201]
[0202] Precursor 1B (14 mg) was treated with 2.5% TFA / water (2.8 mL) under argon for 40 min. The reaction mixture was diluted with water (31 mL) and the product was lyophilized (using isopropanol / dry ice under argon) to yield 18 mg of precursor 2. The lyophilized product was analyzed by LC-MS (gradient: 20-50% B in 5 minutes, t R : 2.5 and 2.1 minutes, measured values m / z : 1043.8, expected MH 2 2+ : 1043.5) analysis.
[0203] Chromatographic resolution of precursor 2 regioisomers can be achieved on analytical HPLC using 0.1% TFA. However, separation of the regioisomers was not attempted at this stage due to the high reactivity of free aminooxy groups towards traces of ketones and aldehydes in solvent and atmosphere.
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