Pretargeting kit, method and agents used therein
A kit, pre-targeting technology, used in nuclear imaging and radiotherapy, pre-targeting reagents, pre-targeting kits, non-bioreactive chemical groups, can solve complex chemical problems
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Embodiment 1
[0119] like Figure 7a Molecule 1 (see Figure 8 ). An example of the corresponding probe 2 (derived from 4-(2-hydroxyethoxy)phenylethylene) is in Figure 9 shown in . Both molecules contain PEG chains. Molecule 1 comprises an N-hydroxysuccinimidyl moiety for coupling the molecule to an amino group present in the antibody. The DOTA-derived moieties in 2 can be used to carry rare earth metal ions such as Gd for MR imaging or Lu-177 for nuclear imaging and therapy (SPECT).
[0120] Figure 8 The synthesis of 1 is shown in . The starting tetrazine-derived molecule 5 was prepared according to Blackman et al. (Blackman, ML; Royzen, M; Fox, JM, Journal of The American Chemical Society, 2008, 130(41), 13518-19). It is converted to acid 6 by reaction with succinic anhydride, followed by its N-hydroxysuccinimidyl ester 7. This N-hydroxysuccinimidyl ester was used to form acid 9 which in turn was converted to its hydroxysuccinimidyl ester 1 by reaction with a commercially availa...
Embodiment 2
[0123] Compared to Example 1, this example illustrates the opposite molecular pair, namely 1) a 4-(2-hydroxy)ethoxy)phenylethylene derivative that will form the pre-targeting moiety after attachment to the antibody 3 and, 2) Probe 4 derived from said tetrazine / DOTA, which can be used as Figure 7b Effector probes shown in , respectively, in Figure 10 and 11 shown in .
[0124] Commercially available (IRIS biochem) PEG derivative 8 with N-hydroxysuccinimidyl ester 14 (see Figure 9 ) to form acid 19, followed by the formation of N-hydroxysuccinimidyl derivatives derived from this acid to form 4-(2-hydroxy)ethoxy)phenylethylene derivatives 3.
[0125] The synthesis of probe 4 derived from tetrazine / DOTA is shown in FIG. 7 . By amine 18 derived from DOTA and PEG (see Figure 9 ) and N-hydroxysuccinimidyl ester 7 (see Figure 8 ) reaction to prepare this probe.
Embodiment 3
[0126] Example 3: In Vivo Imaging
[0127] All reagents and solvents were obtained from commercial sources (reagents from Sigma-Aldrich, Acros, ABCR, Invitrogen, and Merck, conventional and deuterated solvents from Biosolve, Merck, and Cambridge Isotope Laboratories) and were not further prepared unless otherwise stated. The purification is ready to use. 1-Amino-3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36-dodeoxanonacosane-39-oic acid (S11) and (35-amino-3 , 6, 9, 12, 15, 18, 21, 24, 27, 30, 33-tert-butyl carbamate (S3) from Polypure (Norway) and Iris Biotech (Germany )get. 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10- Tetraazacyclododecane-1,4,7-triyl)triacetic acid (S6) as the HPF 6 and about 3 equivalents of trifluoroacetic acid (TFA) were obtained from Macrocyclics (USA). Rituximab solution (MabThera ) were purchased from Roche (Switzerland). [ 111 In] indium chloride and [ 125 I] Sodium iodide solution was purchased from PerkinElmer (USA...
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