COMPOUNDS AND METHODS FOR PURIFICATION OF SERINE PROTEASES
Disclosed herein are compounds, compositions, methods and kits for purifying a serine protease and serine proteases purified with the compounds, compositions and methods.
1 - 18 . (canceled)
19 . A compound selected from:
or a salt thereof.
20 . (canceled)
21 . An affinity solid support of Formula II:
or a salt thereof,
wherein:
R 21 is —CF 3 , —SO 2 CH 3 , —X-L-Y—Z,
R 22 is —OCH 3 , chloro, or X-L-Y—Z;
R 23 is hydrogen or chloro;
X is a covalent bond, O, S, SO 2 , C(O)NH, NHC(O) or NH;
L-Y—Z is -L 1 -Y—Z,
L 1 -Y—Z is
L 2 -Y—Z is
L 3 -Y—Z is
Y—Z is
Z is Capto™ resin;
q is 1, 2, 3, 4, 5, 6 or 7;
r is 1, 2, 3, 4, 5, 6 or 7;
s is 1, 2, 3, 4, 5, 6 or 7;
t is 1, 2, 3, 4, 5, 6 or 7;
v is 1, 2, 3, 4, 5, 6 or 7;
u is 1, 2, 3, 4, 5, 6 or 7;
w is 1, 2 or 3;
n, m, and p are either 0 or 1, with the provisos that
(1) when R 21 is
—CF 3 or —SO 2 CH 3 , and R 22 is —OCH 3 or chloro, then one of n, m, and p must be 1, and the others of n, m, and p must be zero; and
(2) when R 21 is other than
—CF 3 or —SO 2 CH 3 , or R 22 is X-L-Y—Z, then all of n, m, and p must be zero.
22 . The affinity solid support of claim 21 of Formula II-A:
wherein the variables are as defined in claim 21 .
23 . The affinity solid support of claim 21 of Formula II-B:
wherein the variables are as defined in claim 21 .
24 . The affinity solid support of claim 21 of Formula II-C:
wherein the variables are as defined in claim 21 .
25 . The affinity solid support of claim 21 of Formula II-D:
wherein the variables are as defined in claim 21 .
26 . The affinity solid support of claim 21 , wherein R 21 is
27 . The affinity solid support of claim 21 , wherein R 22 is —OCH 3 .
28 . The affinity solid support of claim 21 , wherein R 22 is X-L-Y—Z.
29 . The affinity solid support of claim 21 , wherein R 23 is hydrogen.
30 . The affinity solid support of claim 21 , wherein R 23 is chloro.
31 . The affinity solid support of claim 21 , wherein X is O.
32 . The affinity solid support of claim 21 , wherein X is a covalent bond.
33 . The affinity solid support of claim 21 , wherein X is S or SO 2 .
34 . The affinity solid support of claim 21 , wherein X is NH.
35 . The affinity solid support of any claim 21 , wherein L-Y—Z is -L 1 -Y—Z or
36 . The affinity solid support of claim 21 , wherein L-Y—Z is selected from the group consisting of
37 . The affinity solid support of claim 21 , wherein L 1 -Y—Z is selected from the group consisting of
38 . The affinity solid support of claim 21 , wherein each of q, r, s, and t is at least 3.
39 . The affinity solid support of claim 21 , wherein —X-L-Y—Z is selected from the group consisting of
40 . The affinity solid support of claim 21 which is
or a salt thereof.
41 . (canceled)
42 . An affinity solid support of formula:
or a salt thereof, wherein v is 1, 2, 3, 4, 5, 6 or 7 and Z is Capto™ resin.
43 - 52 . (canceled)
53 . A kit for purifying a serine protease comprising
(1) an affinity solid support of any claim 21 , and
(2) an elution buffer comprising a competitive agent.
54 . (canceled)
55 . A kit for purifying a serine protease comprising a compound of Formula I-D:
or a salt thereof,
and a resin of the Formula
wherein
R 1 is —CF 3 , —SO 2 CH 3 , —X-L-R,
R 2 is —OCH 3 , chloro, or X-L-R;
R 3 is hydrogen or chloro;
X is a covalent bond, O, S, SO 2 , C(O)NH, NHC(O) or NH;
L-R is —(CH 2 ) w —CO 2 H;
R is CO 2 H;
n, m, and p are either 0 or 1, with the provisos that
(1) when R 1 is
—CF 3 or —SO 2 CH 3 , and R 2 is —OCH 3 or chloro, then one of n, m, and p must be 1, and the others of n, m, and p must be zero; and
(2) when R 1 is other than
—CF 3 or —SO 2 CH 3 , or R 2 is X-L-R, then all of n, m, and p must be zero;
u is 1, 2, 3, 4, 5, 6 or 7, w is 1, 2 or 3 and Z is Capto™ resin.
56 . The kit of claim 54 , further comprising an elution buffer comprising a competitive agent.
57 . The kit of claim 54 , further comprising a washing buffer.
58 - 59 . (canceled)