Background
Methods
General
Synthesis of the 2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (10)
Synthesis of 3-(2-(2-(2-fluoroethoxy)ethoxy)ethoxy)prop-1-yne (9)
Synthesis of γ-(11-azido-3,6,9-trioxaundecanyl)folic acid amide (5)
Relative lipophilicity (k’ value)
In vitro binding affinity assays
In vitro metabolite studies in fetal calf serum
In vivo studies
Results and discussion
Organic chemistry
Cu(I)I (%) | Cu(I)I + sodium | Cu(II)acetate + sodium | Cu(II)sulfate + sodium | |
---|---|---|---|---|
ascorbate (%) | ascorbate (%) | ascorbate (%) | ||
Microwavea
| 60 ± 3 | 22 | 45 | n.d. |
Oil bathb
| 20 | n.d. | 30 | 15 |
Relative lipophilicity (k’ value)
Radiochemistry
In vitro metabolite studies in fetal calf serum
In vitro binding affinity assays
Ex vivo biodistribution studies
30 min p.i. | 60 min p.i. | 90 min p.i. | 60 min p.i. blockade
a
|
18
F-click folate 45 min p.i.
b
| |
---|---|---|---|---|---|
(n = 3) | (n = 3) | (n = 3) | (n = 3) | (n = 4) | |
%ID/g | |||||
Blood | 0.20 ± 0.02 | 0.18 ± 0.01 | 0.16 ± 0.02 | 0.04 ± 0.01 | 0.13 ± 0.01 |
Heart | 1.81 ± 0.61 | 1.09 ± 0.07 | 0.79 ± 0.16 | 0.02 ± 0.01 | 0.90 ± 0.22 |
Lung | 1.21 ± 0.15 | 1.02 ± 0.7 | 0.86 ± 0.17 | 0.04 ± 0.03 | 0.85 ± 0.04 |
Spleen | 0.53 ± 0.14 | 0.49 ± 0.04 | 0.43 ± 0.08 | 0.06 ± 0.03 | 0.33 ± 0.12 |
Kidneys
|
41.80 ± 2.58
|
40.77 ± 4.34
|
41.04 ± 7.04
|
0.26 ± 0.08
|
16.53 ± 2.22
|
Stomach (empty) | 2.18 ± 0.76 | 1.39 ± 0.13 | 0.96 ± 0.37 | 0.12 ± 0.11 | 2.50 ± 0.6 |
Intestines (empty) | 23.52 ± 7.37 | 4.56 ± 2.31 | 1.92 ± 0.52 | 14.72 ± 19.32 | 19.59 ± 5.26 |
Feces | 164.2 ± 72.5 | 29.48 ± 19.04 | 11.36 ± 4.29 | 105.5 ± 128.7 | 56.00 ± 27.64 |
Liver | 5.24 ± 1.57 | 4.05 ± 0.67 | 2.30 ± 0.54 | 0.21 ± 0.09 | 1.71 ± 0.14 |
Gall bladder | 309.3 ± 187.6 | 133.2 ± 67.38 | 55.13 ± n.d. | 775.5 ± 206.5 | 667.4 ± 530.1 |
Muscle | 1.91 ± 0.25 | 1.64 ± 0.46 | 1.17 ± 0.03 | 0.08 ± 0.09 | n.d. |
Bone | 1.47 ± 0.05 | 1.13 ± 0.21 | 0.84 ± 0.16 | 0.06 ± 0.06 | 0.13 ± 0.01 |
Salivary glands | 8.01 ± 1.16 | 9.28 ± 1.68 | 7.03 ± 1.98 | 0.20 ± 0.18 | n.d. |
Tumor
|
3.39 ± 0.54
|
3.39 ± 0.44
|
3.54 ± 0.68
|
0.19 ± 0.07
|
3.13 ± 0.83
|
Ratio of tumor to organ or tissue | |||||
Blood | 16.81 ± 3.13 | 18.95 ± 2.68 | 22.72 ± 6.73 | 24,08 ± 0.82 | |
Liver | 0.69 ± 0.22 | 0.86 ± 0.19 | 1.55 ± 0.15 | 1.18 ± 0.69 | |
Kidney
|
0.08 ± 0.02
|
0.08 ± 0.01
|
0.09 ± 0.01
|
0.19 ± 1.38
|