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. Author manuscript; available in PMC: 2008 Sep 15.
Published in final edited form as: J Med Chem. 2006 Jun 15;49(12):3536–3543. doi: 10.1021/jm060240a

Table 1.

Physicochemical and biological characterization of GnRH antagonists

No. Compound Purity tRc (min) MSd (M + H)+ pIC50e IC50f (nM) Duration of actiong
HPLCa CZEb Calc. Obs. Avg. ± SEM
1 [Ac-d-2Nal1,d-4Cpa2,d-3Pal3,4Aph(l-Hor)5,d-4Aph(Cbm)6,Ilys8,d-Ala10]GnRH (degarelix) 96 98 23.8 1631.8 1631.9 9.24 ± 0.16 0.58 Very long
2 [Ac-d-2Nal1,d--4Cpa2,d-3Pal3, 4Aph(Ac)5,d-4Aph(Ac)6,Ilys8,d-Ala10]GnRH (acyline) 98 98 28.2 1532.7 1532.7 9.16 ± 0.06 0.69 Long
3 [d-Dap(CO-NH-OH)3]degarelix 97 98 21.7 1628.7 1628.7 8.46 ± 0.01 3.46 Short
4 [d-Dab(CO-NH-OH)3]degarelix 99 99 21.3 1642.8 1642.8 8.57 ± 0.12 2.68 Short
5 [d-Dap(CO-NH-OCH3)3]degarelix 95 99 23.3 1642.8 1642.7 8.50 ± 0.02 3.16 Short
6 [d-Dab(CO-NH-OCH3)3]degarelix 98 98 22.8 1656.8 1656.8 8.77 ± 0.05 1.69 Short
7 [4Aph(CO-NH-OH)5]degarelix 99 98 24.4 1550.7 1550.7 9.07 ± 0.19 0.86 Short
8 [4Aph(CO-NH-OCH3)5]degarelix 99 99 26.3 1564.7 1564.7 9.22 ± 0.03 0.60 Long
9 [4Aph(CO-NH-(C2H4O)2-C2H5)5]degarelix 98 98 29.1 1650.8 1650.8 8.50 ± 0.62 3.16 Long*
10 [4Aph(CO-(C2H4O)3-C2H4-NH-CO-NH-OCH3)5]degarelix 98 97 26.1 1767.9 1767.9 8.97 ± 0.13 1.06 Short
11 des-Ac-[4Aph(CO-NH-OCH3)5]-CH3-(CH2)2-CO-degarelix 98 97 30.1 1592.8 1592.7 8.83 ± 0.31 1.46 Long*
12 des-Ac-[4Aph(CO-NH-OCH3)5]-CH3-(CH2)6-CO-degarelix 97 98 40.4 1648.8 1648.6 8.38 ± 0.14 4.17 Short
13 des-Ac-[4Aph(CO-NH-OCH3)5]-C2H5-(C2H4O)2-NH-CO-degarelix 96 96 30.6 1681.8 1681.9 8.53 ± 0.47 2.98 Long*
14 des-Ac-[4Aph(CO-NH-(C2H4O)2-C2H5)5]-C2H5-(C2H4O)2-NH-CO-degarelix 95 95 33.3 1767.9 1767.9 8.95 ± 0.04 1.12 Short
15 [d-4Aph(CO-NH-OH)6]degarelix 97 99 23.7 1647.7 1647.7 9.09 ± 0.12 0.80 Long*
16 [d-4Aph(CO-NH-OCH3)6]degarelix 98 99 26.0 1661.8 1661.8 9.29 ± 0.12 0.51 Long*
17 [d-4Aph(CO-NH-(C2H4O)2-C2H5)6]degarelix 98 98 28.3 1747.8 1747.9 9.06 ± 0.11 0.88 Short
18 des-Ac-[d-4Aph(CO-NH-OCH3)6]-CH3-(CH2)2-CO-degarelix 98 98 29.7 1689.8 1689.9 8.67 ± 0.26 2.13 Short
19 des-Ac-[d-4Aph(CO-NH-OCH3)6]-CH3-(CH2)6-CO-degarelix 98 98 39.6 1745.9 1745.8 8.02 ± 0.18 9.47 Short
20 des-Ac-[d-4Aph(CO-NH-OCH3)6]-C2H5-(C2H4O)2-NH-CO-degarelix 95 99 29.9 1778.8 1778.7 8.79 ± 0.25 1.61 Short
21 des-Ac-[d-4Aph(CO-NH-(C2H4O)2-C2H5)6]-C2H5-(C2H4O)2-NH-CO-degarelix 95 99 32.1 1864.9 1864.9 8.34 ± 0.37 4.55 ND
22 [4Aph(CO-NH-OCH3)5,d-4Aph(CO-NH-OCH3)6]degarelix 95 96 28.5 1594.8 1594.8 9.04 ± 0.01 0.91 Long*
23 [Nα-Me-4Aph(CO-NH-OCH3)5,d-4Aph(CO-NH-OCH3)6]degarelix 99 98 28.9 1608.8 1608.8 9.15 ± 0.05 0.70 Short
24 [4Aph(CO-NH-(C2H4O)2-C2H5)5,d-4Aph(CO-NH-(C2H4O)2-C2H5)6]degarelix 96 99 33.5 1767.0 1767.1 8.69 ± 0.31 2.04 Short
25 [4Aph(CO-NH-OCH3)5,d-4Aph(CO-(C2H4O)6-C2H4-NH-CO-NH-OCH3)6]degarelix 98 98 28.5 1929.9 1929.8 8.78 ± 0.24 1.67 Short
26 des-Ac-[4Aph(CO-NH-OCH3)5,d-4Aph(CO-NH-OCH3)6]-CH3-(CH2)2-CO-degarelix 99 99 32.3 1622.8 1622.9 8.86 ± 0.35 1.37 Short
27 des-Ac-[4Aph(CO-NH-OCH3)5,d-4Aph(CO-NH-OCH3)6]-CH3-(CH2)6-CO-degarelix 99 99 42.5 1678.8 1678.9 8.30 ± 0.34 4.99 Inactive
28 des-Ac-[4Aph(CO-NH-(C2H4O)2-C2H5)5,d-4Aph(CO-NH-(C2H4O)2-C2H5)6]-C2H5-(C2H4O)2-NH-CO-degarelix 95 97 37.6 1883.9 1883.9 8.59 ± 0.22 2.55 Short
29 [d-Gln3, 4Aph(CO-NH-OCH3)5,d-4Aph(CO-NH-OCH3)6]degarelix 99 99 27.6 1574.7 1574.7 8.53 ± 0.24 2.98 Short
30 [d-Dab(CO-NH-OCH3)3, 4Aph(CO-NH-OCH3)5,d-4Aph(CO-NH-OCH3)6]degarelix 98 98 28.9 1619.6 1619.8 8.83 ± 0.04 1.49 Short
31 [4Aph(CO-NH-OCH3)5,d-4Aph(CO-NH-OCH3)6,Orn(IGly)8]degarelix 99 99 30.9 1637.8 1637.7 8.82 ± 0.20 1.53 Short
a

Percentage purity determined by HPLC using buffer system A; TEAP, pH 2.30, buffer system B, 60% CH3CN/40% A under gradient conditions (30% to 80% B over 50 min), at flow rate of 0.2 mL/min on a Vydac C18 column (0.21 × 15 cm, 5 μm particle size, 300 Å pore size). Detection at 214 nm.

b

Percentage purity determined by capillary zone electrophoresis (CZE ) using a Beckman P/ACE System 2050 controlled by an IBM Personal system/2 model 50Z; field strength of 15 kV at 30 °C. Buffer, 100 mM sodium phosphate (85:15, H2O:CH3CN), pH 2.50, on a Agilent μSil bare fused-silica capillary (75 μm i.d. × 40 cm length). Detection at 214 nm.

c

Retention times under gradient conditions (30% to 80% B over 50 min); buffer system A; TEAP, pH 7.0, buffer system B, 60% CH3CN/40% A.

d

Mass spectra (MALDI-MS) were measured on an ABI-Voyager DESTR instrument using saturated solution of α-cyano-4-hydroxycinnamic acid in 0.3% trifluoroacetic acid and 50% acetonitrile as matrix. The calculated [M + H]+of the monoisotope compared with the observed [M + H]+ monoisotopic mass.

e

The pIC50 is the negative log of the IC50 in Molar, as determined in the GnRH reporter gene assay.

f

IC50 is the concentration of antagonist required to repress the GnRH induced response by 50% in the reporter gene assay in HEK-293 cells expressing the human GnRH receptor and a GnRH-responsive stably integrated luciferase reporter gene.

g

Castrated male rat assay. Duration of action: very long = over 80% inhibition of LH release for more than 120 h;

*long = over 80% inhibition of LH release at 96 h but not at 120 h; long = over 80% inhibition of LH release at 72 h but not at 96 h; short = over 80% inhibition of LH release at 3 h but not at 72 h; ND = not determined.