U.S. patent application number 14/530444 was filed with the patent office on 2015-02-26 for compositions for treating a disease or condition associated with abnormal angiogenesis.
The applicant listed for this patent is Covx Technologies Ireland Ltd.. Invention is credited to Abhijit Suresh BHAT, Curt William BRADSHAW, Venkata R. DOPPALAPUDI, Jing-Yu LAI, Dingguo LIU.
Application Number | 20150057431 14/530444 |
Document ID | / |
Family ID | 39364902 |
Filed Date | 2015-02-26 |
United States Patent
Application |
20150057431 |
Kind Code |
A1 |
BRADSHAW; Curt William ; et
al. |
February 26, 2015 |
COMPOSITIONS FOR TREATING A DISEASE OR CONDITION ASSOCIATED WITH
ABNORMAL ANGIOGENESIS
Abstract
The present invention provides methods of treating (including
preventing) a disease or condition associated with abnormal
angiogenesis in a subject include administering a therapeutically
effective amount of an AA targeting compound of the invention to
the subject. The AA targeting compounds comprise AA targeting
agent-linker conjugates which are linked to a combining site of an
antibody.
Inventors: |
BRADSHAW; Curt William; (San
Diego, CA) ; BHAT; Abhijit Suresh; (Encinitas,
CA) ; LAI; Jing-Yu; (San Diego, CA) ;
DOPPALAPUDI; Venkata R.; (San Diego, CA) ; LIU;
Dingguo; (San Diego, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Covx Technologies Ireland Ltd. |
Co Cork |
|
IE |
|
|
Family ID: |
39364902 |
Appl. No.: |
14/530444 |
Filed: |
October 31, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13584721 |
Aug 13, 2012 |
8912148 |
|
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14530444 |
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11938766 |
Nov 12, 2007 |
8288349 |
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13584721 |
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60865360 |
Nov 10, 2006 |
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60939830 |
May 23, 2007 |
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60945329 |
Jun 20, 2007 |
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Current U.S.
Class: |
530/326 |
Current CPC
Class: |
A61P 31/00 20180101;
C07K 2319/30 20130101; A61P 35/02 20180101; A61P 9/12 20180101;
A61P 9/00 20180101; A61P 19/02 20180101; C07K 14/515 20130101; A61P
27/02 20180101; A61P 13/12 20180101; A61P 35/00 20180101; A61P
35/04 20180101; C07K 14/47 20130101; A61P 31/04 20180101; A61K
38/00 20130101; A61P 17/06 20180101 |
Class at
Publication: |
530/326 |
International
Class: |
C07K 14/47 20060101
C07K014/47 |
Claims
1. A compound of the formula: R.sup.1-[AA targeting agent]-R.sup.2
wherein R.sup.1 is CH.sub.3, C(O)CH.sub.3, C(O)CH.sub.3,
C(O)CH.sub.2CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.2CH.sub.3, C(O)C.sub.6H.sub.5,
C(O)CH.sub.2CH.sub.2(CH.sub.2CH.sub.2O).sub.1-5Me, dichlorobenzoyl
(DCB), difluorobenzoyl (DFB), pyridinyl carboxlate (PyC) or
amido-2-PEG, an amino protecting group, a lipid fatty acid group or
a carbohydrate; and R.sup.2 is OH, NH.sub.2, NH(CH.sub.3),
NHCH.sub.2CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.2CH.sub.3, NHC.sub.6H.sub.5,
NHCH.sub.2CH.sub.2OCH.sub.3, NHOCH.sub.3, NHOCH.sub.2CH.sub.3, a
carboxy protecting group, a lipid fatty acid group or a
carbohydrate, and wherein [AA targeting agent] comprises a peptide
of the sequence: TABLE-US-00030 (SEQ ID NO: 193)
R.sup.1-Q.sup.1x.sup.2Y.sup.3Q.sup.4x.sup.5L.sup.6D.sup.7x.sup.8x.sup.9D.s-
up.10x.sup.11x.sup.12x.sup.13x.sup.14x.sup.15x.sup.16F.sup.17x.sup.18x.sup-
.19Q.sup.20Q.sup.21 G.sup.22-R.sup.2
wherein one of Q.sup.1, x.sup.8, x.sup.9, x.sup.11, x.sup.12,
x.sup.15, x.sup.16, x.sup.18, x.sup.19 or G.sup.22 is a linking
residue comprising a nucleophilic side chain covalently linkable to
the combining site of an antibody directly or via an intermediate
linker, the linking residue being selected from the group
comprising K, R, Y, C, T, S, homologs of lysine, homocysteine,
homoserine, Dap, and Dab, or the N-terminus or C-terminus, and
x.sup.2 is selected from the group consisting of K, N, R, H, AcK,
Nick, CbcK and the linking residue, and x.sup.5 is selected from
the group consisting of P, hP, dhP, or BnHP, and x.sup.9 is
selected from the group consisting of L, I, ThA, AcK and the
linking residue, and x.sup.8 is selected from the group consisting
of E and the linking residue, and x.sup.11 is selected from the
group consisting of Q, N, C, K, AcK, Dab, Dap and the linking
residue, and x.sup.12 is selected from the group consisting of L,
HL, Nva, I, HchA, HF, ThA, and x.sup.13 is selected from the group
consisting of L, HL, Nva, I, HchA, HF, ThA, and x.sup.14 is
selected from the group consisting of aromatic residues and the
linking residue, and x.sup.15 is selected from the group consisting
of D and the linking residue, and x.sup.16 is selected from the
group consisting of Q, N and the linking residue, and x.sup.18 is
selected from the group consisting of M and the linking residue,
and. x.sup.19 is selected from the group consisting of L, I and the
linking residue.
2. A compound as claimed in claim 1, wherein the linking residue is
selected from the group consisting of K, Y, T, Dap and Dab.
3. A compound as claimed in claim 1, wherein the linking residue is
K.
4. A compound as claimed in claim 1, wherein the linking residue is
located at one of x.sup.9, x.sup.11, x.sup.12, x.sup.15, x.sup.16,
x.sup.18 and x.sup.19.
5. A compound as claimed in claim 1, wherein x.sup.11 is the
linking residue.
6. A compound as claimed in claim 1, wherein x.sup.2 is selected
from the group consisting of K, N, and AcK, and x.sup.5 is selected
from the group consisting of P, hP, and dhP, and x.sup.11 is
selected from the group consisting of K, AcK, and x.sup.13 is
selected from the group consisting of L, HL, Nva, I, and x.sup.14
is selected from the group consisting of F, Y, W, BPA, CF, NF, and
x.sup.16 is Q.
7. A compound as claimed in claim 1, wherein x.sup.2 is N.
8. A compound as claimed in claim 1, wherein x.sup.2 is AcK.
9. A compound as claimed in claim 1, wherein x.sup.5 is P.
10. A compound as claimed in claim 1, wherein x.sup.13 is L.
11. A compound as claimed in claim 1, wherein x.sup.14 is
consisting of Y.
12. A compound as claimed in claim 1, wherein x.sup.9 is L.
13. A compound as claimed in claim 1, wherein x.sup.9 is AcK.
14. A compound as claimed as claimed in claim 1, wherein the
peptide comprises a sequence substantially homologous to one or
more compounds selected from the group consisting of: SEQ ID NO:21,
SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID
NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ
ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35,
SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID
NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ
ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49,
SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID
NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ
ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63,
SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID
NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ
ID NO:73, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77,
SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID
NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ
ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90, SEQ ID NO:91,
SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID
NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ
ID NO:101, SEQ ID NO:102, SEQ ID NO:103, SEQ ID NO:104, SEQ ID
NO:105, SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109,
SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO:113, SEQ ID
NO:114, SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, SEQ ID NO:118,
SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:121, SEQ ID NO:122, SEQ ID
NO:123, SEQ ID NO:124, SEQ ID NO:125, SEQ ID NO:126, SEQ ID NO:127,
SEQ ID NO:128, SEQ ID NO:129, SEQ ID NO:130, SEQ ID NO:131, SEQ ID
NO:132, SEQ ID NO:133, SEQ ID NO:134, SEQ ID NO:135, SEQ ID NO:136,
SEQ ID NO:137, SEQ ID NO:138, SEQ ID NO:139, SEQ ID NO:140, SEQ ID
NO:141, SEQ ID NO:142, SEQ ID NO:143, SEQ ID NO:144, SEQ ID NO:145,
SEQ ID NO:146, SEQ ID NO:147, SEQ ID NO:148, SEQ ID NO:149, SEQ ID
NO:150, SEQ ID NO:151, SEQ ID NO:152, SEQ ID NO:153, SEQ ID NO:154,
SEQ ID NO:155, SEQ ID NO:156, SEQ ID NO:157, SEQ ID NO:158, SEQ ID
NO:159, SEQ ID NO:160, SEQ ID NO:161, SEQ ID NO:162, SEQ ID NO:163,
SEQ ID NO:164, SEQ ID NO:165, SEQ ID NO:166, SEQ ID NO:167, SEQ ID
NO:168, SEQ ID NO:169, SEQ ID NO:170, SEQ ID NO:171, SEQ ID NO:172,
SEQ ID NO:173, SEQ ID NO:174, SEQ ID NO:175, SEQ ID NO:176, SEQ ID
NO:177, and SEQ ID NO:178.
15. A peptide at least 90% homologous to a peptide selected from
the group consisting of: SEQ ID NO: 22, SEQ ID NO: 34, SEQ ID NO:
41, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 48 and SEQ ID NO:
64.
16. A peptide according to claim 15, wherein at least 80%
homologous to a peptide selected from the group consisting of: SEQ
ID NO: 22, SEQ ID NO: 34, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO:
44, SEQ ID NO: 48 and SEQ ID NO: 64.
17. A peptide according to claim 15, wherein at least 70%
homologous to a peptide selected from the group consisting of: SEQ
ID NO: 22, SEQ ID NO: 34, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO:
44, SEQ ID NO: 48 and SEQ ID NO: 64.
18. A peptide according to claim 1, selected from the group
consisting of: R.sup.1-Q(AcK)Y QPL DEL DKT LYD QFM LQQ G-R.sup.2;
R.sup.1-QNY QPL DEL DKT LYD QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y QPL
DEK D(AcK)T LYD QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y QPL DEK DET LYD
QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y Q(DHP)L DEL DKT LYD QFM LQQ
G-R.sup.2; R.sup.1-Q(AcK)Y Q(HP)L DEL DKT LYD QFM LQQ G-R.sup.2;
R.sup.1-Q(AcK)Y QPL DEL DKT IYD QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y
QPL DEI DKT LYD QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y QPL DEL DKT
(HChA)YD QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y QPL DEL DKT (HF)YD QFM
LQQ G-R.sup.2; R.sup.1-Q(AcK)Y QPL DEL DKT (ThA)YD QFM LQQ
G-R.sup.2; R.sup.1-Q(AcK)Y QPL DEL DKT (Nva)YD QFM LQQ G-R.sup.2;
R.sup.1-Q(AcK)Y QPL DEL DKT (HL)YD QFM LQQ G-R.sup.2;
R.sup.1-Q(AcK)Y QPL DE(AcK) DKT LYD QFM LQQ G-R.sup.2;
R.sup.1-Q(AcK)Y QPL DEL DKT LFD QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y
Q(HP)L DE(ThA) DKT L(NO2F)D QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y
Q(HP)L DE(ThA) DKT L(BPA)D QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y
Q(HP)L DE(ThA) DKT L(CO2H)FD QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y QPL
DE(AcK) DKT L(NO2F)D QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y QPL DE(AcK)
DKT LFD QFM LQQ G-R.sup.2; DCB-Q(AcK)Y QPL DEL DKT LYD QFM LQQ
G-R.sup.2; DFB-Q(AcK)Y QPL DEL DKT LYD QFM LQQ G-R.sup.2;
PyC-Q(AcK)Y QPL DEL DKT LYD QFM LQQ G-R.sup.2; Amido 2-PEG-Q(AcK)Y
QPL DEL DKT LYD QFM LQQ G-R.sup.2; R.sup.1-Q(CIBnCarbamateK)Y QPL
DEL DKT LYD QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y QPL DEL D(Dab)T LYD
QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y QPL DEL D(Dap)T LYD QFM LQQ
G-R.sup.2; R.sup.1-Q(AcK)Y QPL DEL DKT L(NO2F)D QFM LQQ G-R.sup.2;
R.sup.1-QNY QPL DEL DKT L(BPA)D QFM LQQ G-R.sup.2; R.sup.1-QNY QPL
DEL DKT L(CF)D QFM LQQ G-R.sup.2; R.sup.1-QNY QPL DEL DKT LYD QFM
LQQ G-R.sup.2; R.sup.1-Q(Nick)Y QPL DEL DKT LYD QFM LQQ G-R.sup.2;
R.sup.1-Q(AcK)Y QPL DE(AcK) DKT L(CF)D QFM LQQ G-R.sup.2;
R.sup.1-Q(AcK)Y QPL DE(AcK) DK(OP)T LFD QFM LQQ G-R.sup.2; wherein:
wherein R.sup.1 is CH.sub.3, C(O)CH.sub.3, C(O)CH.sub.3,
C(O)CH.sub.2CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.2CH.sub.3, C(O)C.sub.6H.sub.5,
C(O)CH.sub.2CH.sub.2(CH.sub.2CH.sub.2O).sub.1-5Me, dichlorobenzoyl
(DCB), difluorobenzoyl (DFB), pyridinyl carboxlate (PyC) or
amido-2-PEG, an amino protecting group, a lipid fatty acid group or
a carbohydrate; and R.sup.2 is OH, NH.sub.2, NH(CH.sub.3),
NHCH.sub.2CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.2CH.sub.3, NHC.sub.6H.sub.5,
NHCH.sub.2CH.sub.2OCH.sub.3, NHOCH.sub.3, NHOCH.sub.2CH.sub.3, a
carboxy protecting group, a lipid fatty acid group or a
carbohydrate.
19. A peptide according to claim 1, selected from the group
consisting of: R.sup.1-Q(AcK)Y QPL DELDKTLYD QFM LQQ G-R.sup.2;
R.sup.1-Q(AcK)Y QPL DEL DKTLYD QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y
QPL DELDKTL(NO2F)D QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y QPL
DEKDK(Ac)T LYD QFM LQQ G-R.sup.2; R.sup.1-Q(AcK)Y QPL DEK(Ac) DKLT
LYD QFM LQQ G-R.sup.2; wherein wherein R.sup.1 is CH.sub.3,
C(O)CH.sub.3, C(O)CH.sub.3, C(O)CH.sub.2CH.sub.3,
C(O)CH.sub.2CH.sub.2CH.sub.3, C(O)CH(CH.sub.3)CH.sub.3,
C(O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.2CH.sub.3, C(O)C.sub.6H.sub.5,
C(O)CH.sub.2CH.sub.2(CH.sub.2CH.sub.2O).sub.1-5Me, dichlorobenzoyl
(DCB), difluorobenzoyl (DFB), pyridinyl carboxlate (PyC) or
amido-2-PEG, an amino protecting group, a lipid fatty acid group or
a carbohydrate; and R.sup.2 is OH, NH.sub.2, NH(CH.sub.3),
NHCH.sub.2CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.2CH.sub.3, NHC.sub.6H.sub.5,
NHCH.sub.2CH.sub.2OCH.sub.3, NHOCH.sub.3, NHOCH.sub.2CH.sub.3, a
carboxy protecting group, a lipid fatty acid group or a
carbohydrate.
20. A peptide of the formula: R.sup.1-Q(AcK)Y QPL DEK(Ac)DKLT LYD
QFM LQQ G-R.sup.2; wherein R.sup.1 is C(O)CH.sub.3 and R.sup.2 is
NH.sub.2.
Description
[0001] This is a Continuation of U.S. application Ser. No.
13/584,721 filed Aug. 13, 2012, which is a Divisional of U.S.
application Ser. No. 11/938,766 filed Nov. 12, 2007, now U.S. Pat.
No. 8,288,349 issued on Oct. 16, 2012, which claims the benefit of
U.S. Provisional Application No. 60/865,360, filed Nov. 10, 2006,
U.S. Provisional Application No. 60/939,830, filed May 23, 2007,
and U.S. Provisional Application No. 60/945,329, filed Jun. 20,
2007, the contents of which are incorporated by reference herein in
their entireties.
CROSS REFERENCE TO SEQUENCE LISTING
[0002] This application includes an electronically submitted
sequence listing in .txt format. The .txt file contains a sequence
listing entitled "PC19771CSequence_Listing.txt" created on Oct. 31,
2014 and having a size of 110 KB. The sequence listing contained in
this .txt file is part of the specification and is herein
incorporated by reference in its entirety.
FIELD OF THE DISCLOSURE
[0003] The present invention relates to novel compounds that
possess anti-angiogenic activity and methods of making and using
these compounds.
BACKGROUND
[0004] Angiogenesis is the fundamental process by which new blood
vessels are formed and is essential to a variety of normal body
activities such as reproduction, development and wound repair.
Although angiogenesis is a highly regulated process under normal
conditions, many diseases (characterized as "angiogenic diseases")
are caused or exacerbated by unregulated angiogenesis. For example,
ocular neovascularization has been implicated as the most common
cause of blindness. In certain existing conditions such as
arthritis, newly formed capillary blood vessels invade the joints
and destroy cartilage. In diabetes, new capillaries formed in the
retina invade the vitreous, bleed, and cause blindness. Growth and
metastasis of solid tumors are also angiogenesis-dependent (J.
Folkman, Cancer Res., 46:467-473 (1986), J. Folkman, J. Natl.
Cancer Inst., 82:4-6 (1989)). It has been shown, for example, that
tumors which enlarge to greater than 2 mm obtain their own blood
supply by inducing the growth of new capillary blood vessels. Once
these new blood vessels become embedded in the tumor, they provide
a means for tumor cells to enter the circulation and metastasize to
distant sites such as the liver, lungs, and bones (N. Weidner, et.
al., N. Engl. J. Med., 324:1-8 (1991)).
[0005] Various peptides have been identified that bind to the
angiogenesis related factor angiopoietin-2 ("Ang-2") (Oliner, J. et
al., Cancer Cell, 204(6), 507-516 (2004)). The Ang-2 binding
peptides have been shown to possess anti-angiogenic activity.
[0006] The reference to any art in this specification is not, and
should not be taken as, an acknowledgement of any form or
suggestion that the referenced art forms part of the common general
knowledge.
BRIEF SUMMARY
[0007] The present invention provides anti-angiogenic targeting
compounds (AA targeting compounds) based upon Ang-2 binding
peptides with unique specificity and biological properties which
are useful in many applications. The anti-angiogenic targeting
compounds of the invention are formed by covalently linking a
anti-angiogenic targeting agent to a combining site of an antibody.
Pharmaceutical compositions comprising targeting compounds of the
invention and a pharmaceutically acceptable carrier are also
provided. In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00001 (SEQ ID NO: 161)
x.sup.1x.sup.2x.sup.3x.sup.4x.sup.5x.sup.6Dx.sup.8x.sup.9x.sup.10x.sup.11x-
.sup.12x.sup.13x.sup.14x.sup.15x.sup.16x.sup.17x.sup.18x.sup.19x.sup.20
x.sup.21x.sup.22
wherein x.sup.1 may be any residue, x.sup.2 may be any residue,
x.sup.3 may be any aromatic amino acid residue, x.sup.4 may be any
residue' x.sup.5 may be P, hP, dhP, or BnHP x.sup.6 may be any
residue, x.sup.8 may be D or E, x.sup.9 may be any residue
comprising a nucleophilic or electrophilic side chain, x.sup.10 may
be D or E, x.sup.11 may be any residue comprising a nucleophilic or
electrophilic side chain, x.sup.12 may be any residue' x.sup.13 may
be selected from the group consisting of L, HL, Nva, I, HchA, HF,
and ThA. x.sup.14 may be any aromatic amino acid residue, x.sup.15
may be D or E, x.sup.16 may be any residue' x.sup.17 may be any
aromatic amino acid residue, x.sup.18 may be any residue' x.sup.19
may be any residue' x.sup.20 may be any residue, or may be absent
when x.sup.22 and x.sup.21 are absent, x.sup.21 may be any residue,
or may be absent when x.sup.22 is absent, x.sup.22 may be any
residue, or absent. x.sup.1 may be Q. x.sup.1 may be T. x.sup.1 may
be S. x.sup.1 may be R. x.sup.1 may be H. x.sup.2 may be K. x.sup.2
may be N. x.sup.2 may be R. x.sup.2 may be H. x.sup.2 may be AcK.
x.sup.2 may be Nick. x.sup.2 may be CbcK. In some embodiments,
x.sup.2 may be selected from the group consisting of K, N, R, H,
AcK, Nick, or CbcK. X.sup.2 may be selected from the group
consisting of N, R, H, AcK, Nick, or CbcK. X.sup.2 may be selected
from the group consisting of R, H, AcK, Nick, or CbcK. X.sup.2 may
be selected from the group consisting of R, H, AcK, or CbcK.
x.sup.2 may be selected from the group consisting of R, H, or AcK.
x.sup.3 may be F. x.sup.3 may be Y. x.sup.3 may be W. x.sup.3 may
be BPA. x.sup.3 may be CF. x.sup.3 may be or NF. x.sup.4 may be Q.
x.sup.4 may be M. x.sup.4 may be E. x.sup.4 may be K. x.sup.5 may
be P. x.sup.5 may be HP. x.sup.5 may be DHP. x.sup.5 may be BnHP.
x.sup.6 may be M. x.sup.6 may be L. x.sup.6 may be I. x.sup.8 may
be E. x.sup.8 may be D. x.sup.9 may be L. x.sup.9 may be I. x.sup.9
may be TA. x.sup.9 may be ThA. x.sup.9 may be K. x.sup.9 may be
AcK. x.sup.10 may be E. x.sup.10 may be D. x.sup.11 may be Q.
x.sup.11 may be N. x.sup.11 may be C. x.sup.11 may be K. x.sup.11
may be AcK. x.sup.11 may be Dab. X.sup.11 may be Dap. x.sup.12 may
be T. x.sup.12 may be R. x.sup.12 may be K. x.sup.13 may be L.
x.sup.13 may be HL. x.sup.13 may be Nva. x.sup.13 may be I.
x.sup.13 may be HchA. x.sup.13 may be HF. x.sup.13 may be ThA.
x.sup.13 may be K. In some embodiments, x.sup.13 may be selected
from the group consisting of L, HL, Nva, I, HchA, HF, and ThA. In
some embodiments, x.sup.13 may be selected from the may group
consisting of L, HL, and Nva. x.sup.14 may be F. x.sup.14 may be Y.
x.sup.14 may be W. x.sup.14 may be BPA. x.sup.14 may be CF.
x.sup.14 may be or NF. x.sup.15 may be D. x.sup.15 may be E.
x.sup.16 may be Q. x.sup.16 may be N. x.sup.17 may be F. x.sup.17
may be Y. x.sup.17 may be W. x.sup.17 may be BPA. X.sup.17 may be
CF. x.sup.17 may be NF. x.sup.18 may be M. x.sup.19 may be L.
x.sup.19 may be I. x.sup.20 may be Q. x.sup.20 may be N. x.sup.21
may be Q. x.sup.21 may be N. x.sup.22 may be G.
[0008] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00002 (SEQ ID NO: 162)
x.sup.1x.sup.2x.sup.3x.sup.4x.sup.5x.sup.6DEx.sup.9Dx.sup.11x.sup.12x.sup-
.13x.sup.14Dx.sup.16x.sup.17x.sup.18x.sup.19x.sup.20
x.sup.21x.sup.22
wherein each of x.sup.1, x.sup.2, x.sup.4, x.sup.6, x.sup.12,
x.sup.16, x.sup.18, x.sup.19 may individually be any residue, each
of x.sup.3 x.sup.14 and x.sup.17 may individually be any aromatic
amino acid residue, x.sup.5 may be one of P, hP, dhP, or BnHP each
of x.sup.9 and x.sup.11 may individually be any residue comprising
a nucleophilic or electrophilic side chain, x.sup.13 may be
selected from the group consisting of L, HL, Nva, I, HchA, HF, and
ThA. x.sup.20 may be any residue, or may be absent when x.sup.22
and x.sup.21 are absent, x.sup.21 may be any residue, or may be
absent when x.sup.22 is absent, x.sup.22 may be any residue, or
absent. x.sup.1 may be Q. x.sup.1 may be T. x.sup.1 may be S.
x.sup.1 may be R. x.sup.1 may be H. x.sup.2 may be K. x.sup.2 may
be N. x.sup.2 may be R. x.sup.2 may be H. x.sup.2 may be AcK.
x.sup.2 may be Nick. x.sup.2 may be CbcK. In some embodiments,
x.sup.2 may be selected from the group consisting of K, N, R, H,
AcK, Nick, or CbcK. x.sup.2 may be selected from the group
consisting of N, R, H, AcK, Nick, or CbcK. X.sup.2 may be selected
from the group consisting of R, H, AcK, Nick, or CbcK. x.sup.2 may
be selected from the group consisting of R, H, AcK, or CbcK.
x.sup.2 may be selected from the group consisting of R, H, or AcK.
x.sup.3 may be F. x.sup.3 may be Y. x.sup.3 may be W. x.sup.3 may
be BPA. x.sup.3 may be CF. x.sup.3 may be NF. x.sup.4 may be Q.
x.sup.4 may be M. x.sup.4 may be E. x.sup.4 may be K. x.sup.5 may
be P. x.sup.5 may be HP. x.sup.5 may be DHP. x.sup.5 may be BnHP.
x.sup.6 may be M. x.sup.6 may be L. x.sup.6 may be I. x.sup.9 may
be L. x.sup.9 may be I. x.sup.9 may be TA. x.sup.9 may be ThA.
x.sup.9 may be K. x.sup.9 may be AcK. x.sup.11 may be Q. x.sup.11
may be N. x.sup.11 may be C. X.sup.11 may be K. x.sup.11 may be
AcK. x.sup.11 may be Dab. X.sup.11 may be Dap. x.sup.12 may be T.
x.sup.12 may be R. x.sup.12 may be K. x.sup.13 may be L. x.sup.13
may be HL. x.sup.13 may be Nva. x.sup.13 may be I. x.sup.13 may be
HchA. x.sup.13 may be HF. x.sup.13 may be ThA. In some embodiments,
x.sup.13 may be selected from the group consisting of L, HL, Nva,
I, HchA, HF, and ThA. In some embodiments, x.sup.13 may be selected
from the group consisting of L, HL, and Nva. x.sup.14 may be F.
x.sup.14 may be Y. x1.sup.4 may be W. x.sup.14 may be BPA. x.sup.14
may be CF. x.sup.14 may be NF. x.sup.16 may be Q. x.sup.16 may be
N. x.sup.17 may be F. x.sup.17 may be Y. x.sup.17 may be W.
x.sup.17 may be BPA. x.sup.17 may be CF. x.sup.17 may be NF.
x.sup.18 may be M. x.sup.19 may be L. x.sup.19 may be I. x.sup.20
may be Q. x.sup.20 may be N. x.sup.21 may be Q. x.sup.21 may be N.
x.sup.22 may be G.
[0009] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00003 (SEQ ID NO: 163)
x.sup.1AcKx.sup.3x.sup.4x.sup.5x.sup.6DEx.sup.9Dx.sup.11x.sup.12x.sup.13x-
.sup.14Dx.sup.16x.sup.17x.sup.18x.sup.19x.sup.20
x.sup.21x.sup.22
wherein each of x.sup.1, x.sup.4, x.sup.6, x.sup.12, x.sup.16,
x.sup.18, x.sup.19 may individually be any residue, each of x.sup.3
x.sup.14 and x.sup.17 may individually be any aromatic amino acid
residue, x.sup.5 may be P, hP, dhP, or BnHP. each of x.sup.9 and
x.sup.11 may individually be any residue comprising a nucleophilic
or electrophilic side chain, x.sup.13 may be selected from the
group consisting of L, HL, Nva, HchA, HF, and ThA, x.sup.20 may be
any residue, or may be absent when x.sup.22 and x.sup.21 are
absent, x.sup.21 may be any residue, or may be absent when x.sup.22
is absent, x.sup.22 may be any residue, or absent. Compounds 22,
24, 29, 30, 31, 32, 33, 44, 45, 56, 57, 58, 19, 20, 21, 22, 23, 24,
25, 26, 27, 28, 29, 30, 31, 32, 34, 35, and 42 in Table 7 exemplify
aspects of the invention covered by this formula. x.sup.1 may be Q.
x.sup.1 may be T. x.sup.1 may be S. x.sup.1 may be R. x.sup.1 may
be H. x.sup.3 may be F. x.sup.3 may be Y. x.sup.3 may be W. x.sup.3
may be BPA. x.sup.3 may be CF. x.sup.3 may be or NF. x.sup.4 may be
Q. x.sup.4 may be M. x.sup.4 may be E. x.sup.4 may be K. x.sup.5
may be P. x.sup.5 may be HP. x.sup.5 may be DHP. x.sup.5 may be
BnHP. x.sup.6 may be M. x.sup.6 may be L. x.sup.6 may be I. x.sup.9
may be L. x.sup.9 may be I. x.sup.9 may be TA. x.sup.9 may be ThA.
x.sup.9 may be K. x.sup.9 may be AcK. x.sup.11 may be Q. x.sup.11
may be N. x.sup.11 may be C. x.sup.11 may be K. x.sup.11 may be
AcK. x.sup.11 may be Dab. x.sup.11 may be Dap. x.sup.12 may be T.
x.sup.12 may be R. x.sup.12 may be K. x.sup.13 may be L. x.sup.13
may be HL. x.sup.13 may be Nva. x.sup.13 may be I. x.sup.13 may be
HchA. x.sup.13 may be HF. x.sup.13 may be ThA. In some embodiments,
x.sup.13 may be selected from the group consisting of L, HL, Nva,
I, HchA, HF, and ThA. In some embodiments, x.sup.13 may be selected
from the group consisting of L, HL, and Nva. x.sup.14 may be F.
x.sup.14 may be Y. x1.sup.4 may be W. x.sup.14 may be BPA. x.sup.14
may be CF. x.sup.14 may be or NF. x.sup.16 may be Q. x.sup.16 may
be N. x.sup.17 may be F. x.sup.17 may be Y. x.sup.17 may be W.
x.sup.17 may be BPA. X.sup.17 may be CF. x.sup.17 may be or NF.
x.sup.18 may be M. x.sup.19 may be L. x.sup.19 may be I. x.sup.20
may be Q. x.sup.20 may be N. x.sup.21 may be Q. x.sup.21 may be N.
x.sup.22 may be G.
[0010] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00004 (SEQ ID NO: 164)
x.sup.1Nx.sup.3x.sup.4x.sup.5x.sup.6DEx.sup.9Dx.sup.11x.sup.12x.sup.13x.su-
p.14Dx.sup.16x.sup.17x.sup.18x.sup.19x.sup.20x.sup.21x.sup.22
wherein each of x.sup.1, x.sup.4, x.sup.6, x.sup.12, x.sup.16,
x.sup.18, x.sup.19 may individually be any residue, each of x.sup.3
x.sup.14 and x.sup.17 may individually be any aromatic amino acid
residue, x.sup.5 may be P, hP, dhP, or BnHP, each of x.sup.9 and
x.sup.11 may individually be any residue comprising a nucleophilic
or electrophilic side chain, x.sup.13 may be selected from the
group consisting of L, HL, Nva, HchA, HF, and ThA, x.sup.20 may be
any residue, or may be absent when x.sup.22 and x.sup.21 are
absent, x.sup.21 may be any residue, or may be absent when x.sup.22
is absent, x.sup.22 may be any residue, or absent. Compounds 23,
25, 28, 56, 57, and 58 in Table 7 exemplify these aspects of the
invention. x.sup.1 may be Q. x.sup.1 may be T. x.sup.1 may be S.
x.sup.1 may be R. x.sup.1 may be H. x.sup.3 may be F. x.sup.3 may
be Y. x.sup.3 may be W. x.sup.3 may be BPA. x.sup.3 may be CF.
x.sup.3 may be or NF. x.sup.4 may be Q. x.sup.4 may be M. x.sup.4
may be E. x.sup.4 may be K. x.sup.5 may be P. x.sup.5 may be HP.
x.sup.5 may be DHP. x.sup.5 may be BnHP. x.sup.6 may be M. x.sup.6
may be L. x.sup.6 may be I. x.sup.9 may be L. x.sup.9 may be I.
x.sup.9 may be TA. x.sup.9 may be ThA. x.sup.9 may be K. x.sup.9
may be AcK. x.sup.11 may be Q. x.sup.11 may be N. x.sup.11 may be
C. x.sup.11 may be K. x.sup.11 may be AcK. x.sup.11 may be Dab.
X.sup.11 may be Dap. x.sup.12 may be T. x.sup.12 may be R. x.sup.12
may be K. x.sup.13 may be L. x.sup.13 may be HL. x.sup.13 may be
Nva. x.sup.13 may be I. x.sup.13 may be HchA. x.sup.13 may be HF.
x.sup.13 may be ThA. In some embodiments, x.sup.13 may be selected
from the group consisting of L, HL, Nva, I, HchA, HF, and ThA. In
some embodiments, x.sup.13 may be selected from the group
consisting of L, HL, and Nva. x.sup.14 may be F. x.sup.14 may be Y.
x1.sup.4 may be W. x.sup.14 may be BPA. X.sup.14 may be CF.
x.sup.14 may be or NF. x.sup.16 may be Q. x.sup.16 may be N.
x.sup.17 may be F. x.sup.17 may be Y. x.sup.17 may be W. x.sup.17
may be BPA. X.sup.17 may be CF. x.sup.17 may be or NF. x.sup.18 may
be M. x.sup.19 may be L. x.sup.19 may be I. x.sup.20 may be Q.
x.sup.20 may be N. x.sup.21 may be Q. x.sup.21 may be N. x.sup.22
may be G.
[0011] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00005 (SEQ ID NO: 165)
x.sup.1x.sup.2x.sup.3x.sup.4x.sup.5x.sup.6DEx.sup.9DKx.sup.12x.sup.13x.sup-
.14Dx.sup.16x.sup.17x.sup.18x.sup.19x.sup.20x.sup.21x.sup.22
wherein each of x.sup.1, x.sup.2, x.sup.4, x.sup.6, x.sup.12,
x.sup.16, x.sup.18, x.sup.19 may individually be any residue, each
of x.sup.3 x.sup.14 and x.sup.17 may individually be any aromatic
amino acid residue, x.sup.5 may be P, hP, dhP, or BnHP x.sup.9 and
may be any residue, and may be selected from the group L, I, K,
ThA, and AcK, x.sup.13 may be selected from the group consisting of
L, HL, Nva, I, HchA, HF, and ThA, x.sup.20 may be any residue, or
may be absent when x.sup.22 and x.sup.21 are absent, x.sup.21 may
be any residue, or may be absent when x.sup.22 is absent, x.sup.22
may be any residue, or absent. x.sup.1 may be Q. x.sup.1 may be T.
x.sup.1 may be S. x.sup.1 may be R. x.sup.1 may be H. x.sup.2 may
be K. x.sup.2 may be N. x.sup.2 may be R. x.sup.2 may be H. x.sup.2
may be AcK. x.sup.2 may be Nick. x.sup.2 may be CbcK. In some
embodiments, x.sup.2 may be selected from the group consisting of
K, N, R, H, AcK, Nick, or CbcK. X.sup.2 may be selected from the
group consisting of N, R, H, AcK, Nick, or CbcK. X.sup.2 may be
selected from the group consisting of R, H, AcK, Nick, or CbcK.
X.sup.2 may be selected from the group consisting of R, H, AcK, or
CbcK. X.sup.2 may be selected from the group consisting of R, H, or
AcK. x.sup.3 may be F. x.sup.3 may be Y. x.sup.3 may be W. x.sup.3
may be BPA. x.sup.3 may be CF. x.sup.3 may be or NF. x.sup.4 may be
Q. x.sup.4 may be M. x.sup.4 may be E. x.sup.4 may be K. x.sup.5
may be P. x.sup.5 may be HP. x.sup.5 may be DHP. x.sup.5 may be
BnHP. x.sup.6 may be M. x.sup.6 may be L. x.sup.6 may be I. x.sup.9
may be L. x.sup.9 may be I. x.sup.9 may be TA. x.sup.9 may be ThA.
x.sup.9 may be K. x.sup.9 may be AcK. x.sup.12 may be T. x.sup.12
may be R. x.sup.12 may be K. x.sup.13 may be L. x.sup.13 may be HL.
x.sup.13 may be Nva. x.sup.13 may be I. x.sup.13 may be HchA.
x.sup.13 may be HF. x.sup.13 may be ThA. In some embodiments,
x.sup.13 may be selected from the group consisting of L, HL, Nva,
I, HchA, HF, and ThA. In some embodiments, x.sup.13 may be selected
from the group consisting of L, HL, and Nva. x.sup.14 may be F.
x.sup.14 may be Y. x1.sup.4 may be W. x.sup.14 may be BPA. X.sup.14
may be CF. x.sup.14 may be or NF. x.sup.16 may be Q. x.sup.16 may
be N. x.sup.17 may be F. x.sup.17 may be Y. x.sup.17 may be W.
x.sup.17 may be BPA. X.sup.17 may be CF. x.sup.17 may be or NF.
x.sup.18 may be M. x.sup.19 may be L. x.sup.19 may be I. x.sup.20
may be Q. x.sup.20 may be N. x.sup.21 may be Q. x.sup.21 may be N.
x.sup.22 may be G.
[0012] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00006
Qx.sup.2x.sup.3Qx.sup.5LDEx.sup.9Dx.sup.11Tx.sup.13x.sup.14DQx.sup.17MLQQ-
G (SEQ ID NO: 166)
wherein x.sup.2 may be AcK, N, R, H, Nick, CbcK, each of x.sup.3
x.sup.14 and x.sup.17 may individually be any aromatic amino acid
residue, x.sup.5 may be P, hP, dhP, or BnHP, x.sup.9 may be any one
of K, AcK, Ile, L or ThA, x.sup.11 may be any one of K, Dab, Dap,
AcK, C, or R, x.sup.13 may be selected from the group consisting of
L, HL, Nva, I, HchA, HF, and ThA. x.sup.2 may be K. x.sup.2 may be
N. x.sup.2 may be R. x.sup.2 may be H. x.sup.2 may be AcK. x.sup.2
may be Nick. x.sup.2 may be CbcK. In some embodiments, x.sup.2 may
be selected from the group consisting of K, N, R, H, AcK, Nick, or
CbcK. X.sup.2 may be selected from the group consisting of N, R, H,
AcK, Nick, or CbcK. X.sup.2 may be selected from the group
consisting of R, H, AcK, Nick, or CbcK. X.sup.2 may be selected
from the group consisting of R, H, AcK, or CbcK. X.sup.2 may be
selected from the group consisting of R, H, or AcK. x.sup.3 may be
F. x.sup.3 may be Y. x.sup.3 may be W. x.sup.3 may be BPA. x.sup.3
may be CF. x.sup.3 may be NF. x.sup.5 may be P. x.sup.5 may be HP.
x.sup.5 may be DHP. x.sup.5 may be BnHP. x.sup.9 may be L. x.sup.9
may be I. x.sup.9 may be TA. x.sup.9 may be ThA. x.sup.9 may be K.
x.sup.9 may be AcK. x.sup.11 may be Q. x.sup.11 may be N. x.sup.11
may be C. x.sup.11 may be K. x.sup.11 may be AcK. x.sup.11 may be
Dab. x.sup.13 may be L. x.sup.13 may be HL. x.sup.13 may be Nva.
x.sup.13 may be I. x.sup.13 may be HchA. x.sup.13 may be HF.
x.sup.13 may be ThA. In some embodiments, x.sup.13 may be selected
from the group consisting of L, HL, Nva, I, HchA, HF, and ThA. In
some embodiments, x.sup.13 may be selected from the group
consisting of L, HL, and Nva. x.sup.14 may be F. x.sup.14 may be Y.
x1.sup.4 may be W. x.sup.14 may be BPA. X.sup.14 may be CF.
x.sup.14 may be or NF. x.sup.17 may be F. x.sup.17 may be Y.
x.sup.17 may be W. x.sup.17 may be BPA. X.sup.17 may be CF.
x.sup.17 may be or NF.
[0013] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00007
Qx.sup.2x.sup.3Qx.sup.5LDEx.sup.9DKTx.sup.13x.sup.14DQx.sup.17MLQQG
(SEQ ID NO: 167)
wherein x.sup.2 may be any one of K, N, R, H, AcK, Nick, or CbcK.
each of x.sup.3 x.sup.14 and x.sup.17 may individually be any
aromatic amino acid residue, x.sup.5 may be P, hP, dhP, or BnHP
x.sup.9 may be any one of K, AcK, Ile, L or ThA, x.sup.13 may be
selected from the group consisting of L, HL, Nva, I, HchA, HF, and
ThA. x.sup.2 may be K. x.sup.2 may be N. x.sup.2 may be R. x.sup.2
may be H. x.sup.2 may be AcK. x.sup.2 may be Nick. x.sup.2 may be
CbcK. In some embodiments, x.sup.2 may be selected from the group
consisting of K, N, R, H, AcK, Nick, or CbcK. x.sup.2 may be
selected from the group consisting of N, R, H, AcK, Nick, or CbcK.
X.sup.2 may be selected from the group consisting of R, H, AcK,
Nick, or CbcK. X.sup.2 may be selected from the group consisting of
R, H, AcK, or CbcK. X.sup.2 may be selected from the group
consisting of R, H, or AcK. x.sup.3 may be F. x.sup.3 may be Y.
x.sup.3 may be W. x.sup.3 may be BPA. x.sup.3 may be CF. x.sup.3
may be or NF. x.sup.5 may be P. x.sup.5 may be HP. x.sup.5 may be
DHP. x.sup.5 may be BnHP. x.sup.9 may be L. x.sup.9 may be I.
x.sup.9 may be TA. x.sup.9 may be ThA. x.sup.9 may be K. x.sup.9
may be AcK. x.sup.13 may be L. x.sup.13 may be HL. x.sup.13 may be
Nva. x.sup.13 may be I. x.sup.13 may be HchA. x.sup.13 may be HF.
x.sup.13 may be ThA. In some embodiments, x.sup.13 may be selected
from the group consisting of L, HL, Nva, I, HchA, HF, and ThA. In
some embodiments, x.sup.13 may be selected from the group
consisting of L, HL, and Nva. x.sup.14 may be F. x.sup.14 may be Y.
x1.sup.4 may be W. x.sup.14 may be BPA. X.sup.14 may be CF.
x.sup.14 may be or NF. x.sup.17 may be F. x.sup.17 may be Y.
x.sup.17 may be W. x.sup.17 may be BPA. X.sup.17 may be CF.
x.sup.17 may be or NF.
[0014] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00008
QAcKx.sup.3Qx.sup.5LDEx.sup.9DKTx.sup.13x.sup.14DQx.sup.17MLQQG
(SEQ ID NO: 168)
wherein each of x.sup.3 x.sup.14 and x.sup.17 may individually be
any aromatic amino acid residue, x.sup.5 may be P, hP, dhP, or BnHP
x.sup.9 may be any one of K, AcK, Ile, L or ThA, x.sup.13 may be
selected from the group consisting of L, HL, Nva, I, HchA, HF, and
ThA. x.sup.3 may be F. x.sup.3 may be Y. x.sup.3 may be W. x.sup.3
may be BPA. x.sup.3 may be CF. x.sup.3 may be or NF. x.sup.5 may be
P. x.sup.5 may be HP. x.sup.5 may be DHP. x.sup.5 may be BnHP.
x.sup.9 may be L. x.sup.9 may be I. x.sup.9 may be TA. x.sup.9 may
be ThA. x.sup.9 may be K. x.sup.9 may be AcK. x.sup.13 may be L.
x.sup.13 may be HL. x.sup.13 may be Nva. x.sup.13 may be I.
x.sup.13 may be HchA. x.sup.13 may be HF. x.sup.13 may be ThA. In
some embodiments, x.sup.13 may be selected from the group
consisting of L, HL, Nva, I, HchA, HF, and ThA. In some
embodiments, x.sup.13 may be selected from the group consisting of
L, HL, and Nva. x.sup.14 may be F. x.sup.14 may be Y. x1.sup.4 may
be W. x.sup.14 may be BPA. X.sup.14 may be CF. x.sup.14 may be or
NF. x.sup.17 may be F. x.sup.17 may be Y. x.sup.17 may be W.
x.sup.17 may be BPA. X.sup.17 may be CF. x.sup.17 may be or NF.
[0015] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00009
QNx.sup.3Qx.sup.5LDEx.sup.9DKTx.sup.13x.sup.14DQx.sup.17MLQQG (SEQ
ID NO: 169)
Wherein each of x.sup.3 x.sup.14 and x.sup.17 may individually be
any aromatic amino acid residue, x.sup.5 may be P, hP, dhP, or BnHP
x.sup.9 may be any one of K, AcK, Ile, L or ThA x.sup.13 may be
selected from the group consisting of L, HL, Nva, I, HchA, HF, and
ThA.
[0016] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00010 (SEQ ID NO: 170)
x.sup.1x.sup.2x.sup.3x.sup.4x.sup.5x.sup.6DEx.sup.9Dx.sup.11x.sup.12Lx.sup-
.14Dx.sup.16x.sup.17x.sup.18x.sup.19x.sup.20x.sup.21x.sup.22
wherein each of x.sup.1, x.sup.4, x.sup.6, x.sup.12, x.sup.16,
x.sup.18, x.sup.19 may individually be any residue, x.sup.2 may be
AcK, N, R, H, Nick, CbcK, each of x.sup.3 x.sup.14 and x.sup.17 may
individually be any aromatic amino acid residue, x.sup.5 may be one
of P, hP, dhP, or BnHP each of x.sup.9 and x.sup.11 may
individually be any residue comprising a nucleophilic or
electrophilic side chain, x.sup.20 may be any residue, or may be
absent when x.sup.22 and x.sup.21 are absent, x.sup.21 may be any
residue, or may be absent when x.sup.22 is absent, x.sup.22 may be
any residue, or absent.
[0017] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00011 (SEQ ID NO: 171)
x.sup.1AcKx.sup.3x.sup.4x.sup.5x.sup.6DEx.sup.9Dx.sup.11x.sup.12Lx.sup.14D-
x.sup.16x.sup.17x.sup.18x.sup.19x.sup.20x.sup.21x.sup.22
wherein each of x.sup.1, x.sup.4, x.sup.6, x.sup.12, x.sup.16,
x.sup.18, x.sup.19 may individually be any residue, each of x.sup.3
x.sup.14 and x.sup.17 may individually be any aromatic amino acid
residue, x.sup.5 may be P, hP, dhP, or BnHP each of x.sup.9 and
X.sup.11 may individually be any residue comprising a nucleophilic
or electrophilic side chain, x.sup.20 may be any residue, or may be
absent when x.sup.22 and x.sup.21 are absent, x.sup.21 may be any
residue, or may be absent when x.sup.22 is absent, x.sup.22 may be
any residue, or absent.
[0018] Compounds 22, 24, 29, 30, 33, 34, 35, 36, 37, 43, 44, 45,
46, 47, 48, 49, 50, 51, 52, 54, 55, and 62 in Table 7 exemplify
aspects of the invention covered by this formula.
[0019] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00012 (SEQ ID NO: 172)
x.sup.1Nx.sup.3x.sup.4x.sup.5x.sup.6DEx.sup.9Dx.sup.11x.sup.12Lx.sup.14Dx.-
sup.16x.sup.17x.sup.18x.sup.19x.sup.20x.sup.21x.sup.22
wherein each of x.sup.1, x.sup.4, x.sup.6, x.sup.12, x.sup.16,
x.sup.18, x.sup.19 may individually be any residue, each of x.sup.3
x.sup.14 and x.sup.17 may individually be any aromatic amino acid
residue, x.sup.5 may be P, hP, dhP, or BnHP each of x.sup.9 and
x.sup.11 may individually be any residue comprising a nucleophilic
or electrophilic side chain, x.sup.20 may be any residue, or may be
absent when x.sup.22 and x.sup.21 are absent, x.sup.21 may be any
residue, or may be absent when x.sup.22 is absent, x.sup.22 may be
any residue, or absent.
[0020] Compounds 23, 25, 28, 56, 57, and 58 in Table 7 exemplify
aspects of the invention covered by this formula.
[0021] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00013 (SEQ ID NO: 173)
x.sup.1x.sup.2x.sup.3x.sup.4x.sup.5x.sup.6DEx.sup.9DKx.sup.12Lx.sup.14Dx.s-
up.16x.sup.17x.sup.18x.sup.19x.sup.20x.sup.21x.sup.22
wherein each of x.sup.1, x.sup.4, x.sup.6, x.sup.12, x.sup.16,
x.sup.18, x.sup.19 may individually be any residue, x.sup.2 may be
AcK, N, R, H, Nick, CbcK, each of x.sup.3 x.sup.14 and x.sup.17 may
individually be any aromatic amino acid residue, x.sup.5 may be P,
hP, dhP, or BnHP x.sup.9 and may be any residue, and may be
selected from the group L, I, K, ThA, and AcK, x.sup.20 may be any
residue, or may be absent when x.sup.22 and x.sup.21 are absent,
x.sup.21 may be any residue, or may be absent when x.sup.22 is
absent, x.sup.22 may be any residue, or absent.
[0022] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00014
Qx.sup.2x.sup.3Qx.sup.5LDEx.sup.9Dx.sup.11TLx.sup.14DQx.sup.17MLQQG
(SEQ ID NO: 174)
Wherein x.sup.2 may be AcK, N, R, H, Nick, CbcK, each of x.sup.3
x.sup.14 and x.sup.17 may individually be any aromatic amino acid
residue, x.sup.5 may be P, hP, dhP, or BnHP, x.sup.9 may be any one
of K, AcK, Ile, L or ThA, x.sup.11 may be any one of K, Dab, AcK,
C, or R,
[0023] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00015
Qx.sup.2x.sup.3Qx.sup.5LDEx.sup.9DKTLx.sup.14DQx.sup.17MLQQG (SEQ
ID NO: 175)
Wherein x.sup.2 may be any one of K, N, R, H, AcK, Nick, or CbcK,
each of x.sup.3 x.sup.14 and x.sup.17 may individually be any
aromatic amino acid residue, x.sup.5 may be P, hP, dhP, or BnHP,
x.sup.9 may be any one of K, AcK, Ile, L or ThA,
[0024] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00016
QAcKx.sup.3Qx.sup.5LDEx.sup.9DKTLx.sup.14DQx.sup.17MLQQG (SEQ ID
NO: 176)
Wherein, each of x.sup.3 x.sup.14 and x.sup.17 may individually be
any aromatic amino acid residue, x.sup.5 may be P, hP, dhP, or
BnHP, x.sup.9 may be any one of K, AcK, Ile, L or ThA,
[0025] In some embodiments, the present invention provides a
targeting agent comprising a peptide comprising a sequence
substantially homologous to:
TABLE-US-00017
QNx.sup.3Qx.sup.5LDEx.sup.9DKTLx.sup.14DQx.sup.17MLQQG (SEQ ID NO:
177)
Wherein each of x.sup.3 x.sup.14 and x.sup.17 may individually be
any aromatic amino acid residue, x.sup.5 may be P, hP, dhP, or
BnHP, x.sup.9 may be any one of K, AcK, Ile, L or ThA.
[0026] Compounds of the invention may comprise an amino-terminus
(N-terminus) capping group or a carboxy-terminus (C-terminus)
capping group. The N-terminus capping group may be "ac":
C(O)CH.sub.3) (e.g. compounds 21, 22, 23, 24, 25, 27, 28, 29, 30,
31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,
48, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63 in Table 7).
Compounds of the invention may comprise a DCB group as N-terminus
capping group (e.g. compound 49 in Table 7). Compounds of the
invention may comprise a DFB group as N-terminus capping group
(e.g. compound 50 in Table 7). Compounds of the invention may
comprise a PyC group as N-terminus capping group (e.g compound 51
in Table 7). Compounds of the invention may comprise a 2-PEG group
as N-terminus capping group (e.g. compound 52 in Table 7). The
C-terminus capping group may be "am": NH.sub.2.
[0027] In some embodiments, the present invention provide an
Angiopoietin receptor-antagonist (AA) targeting agent comprising a
peptide comprising a sequence substantially homologous to:
TABLE-US-00018 (SEQ ID NO: 178)
Q.sup.1x.sup.2Y.sup.3Q.sup.4x.sup.5L.sup.6D.sup.7E.sup.8x.sup.9D.sup.10x.s-
up.11T.sup.12x.sup.13x.sup.14x.sup.15x.sup.16F.sup.17x.sup.18x.sup.19
Q.sup.20Q.sup.21G.sup.22
wherein x.sup.2 is selected from the group consisting of K, N, R,
H, AcK, Nick, CbcK and a linking residue, and x.sup.5 is selected
from the group consisting of P, hP, dhP, or BnHP, and x.sup.9 is
selected from the group consisting of L, I, ThA, AcK and a linking
residue, and x.sup.11 is selected from the group consisting of Q,
N, C, K, AcK, Dab, Dap and a linking residue, and x.sup.13 is
selected from the group consisting of L, HL, Nva, I, HchA, HF, ThA
and a linking residue, and x.sup.14 is selected from the group
consisting of aromatic residues and a linking residue, and x.sup.15
is selected from the group consisting of D and a linking residue,
and x.sup.16 is selected from the group consisting of Q, N and a
linking residue, and x.sup.18 is selected from the group consisting
of M and a linking residue, and x.sup.19 is selected from the group
consisting of L, I and a linking residue, and wherein one of
Q.sup.1, x.sup.9, x.sup.11, x.sup.13, x.sup.15, x.sup.16, x.sup.18,
x.sup.19 and G.sup.22 is a linking residue comprising a
nucleophilic side chain covalently linkable to the combining site
of an antibody directly or via an intermediate linker, the linking
residue being selected from the group comprising K, R, Y, C, T, and
S, or the N-terminus or C-terminus.
[0028] In some embodiments, the present invention provide an
Angiopoietin receptor-antagonist (AA) targeting agent comprising a
peptide comprising a sequence substantially homologous to:
TABLE-US-00019 (SEQ ID NO: 193)
Q.sup.1x.sup.2Y.sup.3Q.sup.4x.sup.5L.sup.6D.sup.7x.sup.8x.sup.9D.sup.10x.s-
up.11x.sup.12x.sup.13x.sup.14x.sup.15x.sup.16F.sup.17x.sup.18x.sup.19
Q.sup.20Q.sup.21G.sup.22
wherein x.sup.2 is selected from the group consisting of K, N, R,
H, AcK, Nick, CbcK and a linking residue, and x.sup.5 is selected
from the group consisting of P, hP, dhP, or BnHP, and x.sup.8 is E
or a linking residue, x.sup.9 is selected from the group consisting
of L, I, ThA, AcK and a linking residue, and x.sup.11 is selected
from the group consisting of Q, N, C, K, AcK, Dab, Dap and a
linking residue, and x.sup.12 is T or a linking residue, and
x.sup.13 is selected from the group consisting of L, HL, Nva, I,
HchA, HF, ThA, and x.sup.14 is selected from the group consisting
of aromatic residues and a linking residue, and x.sup.15 is
selected from the group consisting of D and a linking residue, and
x.sup.16 is selected from the group consisting of Q, N and a
linking residue, and x.sup.18 is selected from the group consisting
of M and a linking residue, and x.sup.19 is selected from the group
consisting of L, I and a linking residue, and wherein one of
Q.sup.1, x.sup.9, x.sup.8, x.sup.11, x.sup.12, x.sup.15, x.sup.16,
x.sup.18, x.sup.19 and G.sup.22 is a linking residue comprising a
nucleophilic side chain covalently linkable to the combining site
of an antibody directly or via an intermediate linker, the linking
residue being selected from the group comprising K, R, Y, C, T, S,
Dap, Dab, homologs of S, homologs of C, homologs of K, or the
N-terminus or C-terminus.
[0029] The linking residue may be selected from the group
consisting of K, Y, T, Dap, and Dab. The linking residue may be
located at one of X.sup.9, X.sup.11, X.sup.12, X.sup.15, X.sup.16,
X.sup.18 and X.sup.19. In some embodiments, x.sup.11 is the linking
residue. TABLEs 7 and 8 provide examples of compounds of the
invention covalently linked to the combining site of h38C2 at
different linging residues.
[0030] x.sup.2 may be selected from the group consisting of K, N,
AcK. Exemplary compounds of the invention where x2 is N are 23, 25,
28, 56, 57 and 58. Exemplary compounds of the invention where x2 is
AcK are 22, 24, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,
42, 43, 43(a), 44, 45, 46, 47, 48, 49, 50, 51, 52, 54, 55, 62, 63,
64, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,
82, 83, 84, 85, 86, 87, and 91
[0031] x.sup.5 may be selected from the group consisting of P, hP,
and dhP. Exemplary compounds of the invention where x5 is P are 21,
23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 35, 36, 37, 38, 39, 40, 41,
42, 43, 43(a), 44, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60,
61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,
78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, and 91.
Exemplary compounds of the invention where x5 is HP are 35, 45, 46,
and 47. Exemplary compounds of the invention where x5 is DHP is
34.
[0032] x.sup.8 may be E.
[0033] x.sup.11 may be selected from the group consisting of K,
AcK, Dab and Dap. Exemplary compounds of the invention where
x.sup.11 is AcK are 30 and 32. Exemplary compounds of the invention
where x.sup.11 is Dab are 54. Exemplary compounds of the invention
where x.sup.11 is Dap are 55.
[0034] X.sup.12 may be T.
[0035] x.sup.13 may be selected from the group consisting of K, L,
HL, Nva, and I. Exemplary compounds of the invention where x.sup.13
is K are 31 and 32 Exemplary compounds of the invention where
x.sup.13 is HL are 42. Exemplary compounds of the invention where
x.sup.13 is Nva are 41. Exemplary compounds of the invention where
x.sup.13 is I are 36.
[0036] x.sup.14 may be selected from the group consisting of F, Y,
W, BPA, CF, and NF. Exemplary compounds of the invention where
x.sup.14 is F are 44. Exemplary compounds of the invention where
x.sup.14 is BPA are 46 and 56. Exemplary compounds of the invention
where x.sup.14 is CF are 47, 57 and 62. Exemplary compounds of the
invention where x.sup.14 is NF are 45, 48 and 58.
[0037] AA targeting agents of the invention may comprise a peptide
comprising a sequence substantially homologous to one or more
compounds selected from the group consisting of: 21, 22, 23, 24,
25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,
42, 43, 43(a) 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56,
57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
and 91
[0038] AA targeting agents of the invention may comprise a peptide
comprising a sequence substantially homologous to one or more
compounds selected from the group consisting of: 24, 25, 26, 27,
28, 29, 30, 34, 35, 37, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 54,
55, 56, 57, 58, 59, 60, 61, 62, 75, 76, 77, 78, and 79.
[0039] AA targeting agents of the invention may comprise a peptide
comprising a sequence substantially homologous to one or more
compounds selected from the group consisting of: 24, 25, 27, 28,
29, 30, 34, 35, 37, 41, 42, 43, 44, 45, 46, 47, 48, 56, 57, 58, 60,
62, 75, 76, 77, 78, and 79.
[0040] AA targeting agents of the invention may comprise a peptide
comprising a sequence substantially homologous to one or more
compounds selected from the group consisting of: 22, 34, 41, 43,
44, 48 and 91.
[0041] AA targeting agents of the invention may comprise a peptide
comprising a sequence substantially homologous to one or more
compounds selected from the group consisting of: 27, 28, 29, 43,
45, and 48.
[0042] The compounds of the invention are useful in targeting Ang2
and demonstrate advantageous properties over existing
Ang2-targeting agents. In some aspects of the invention,
advantageous agents and compounds of the invention provide an
attractive balance between half-life and IC50 values.
[0043] In some embodiments of the invention at least one residue of
x.sup.9 and x.sup.11 is a linking residue comprising an amino acid
whose side chain is capable covalently bonding to chemical groups
comprising electrophiles or nucleophiles respectively.
[0044] The presence of the linking residue provides the targeting
agents of the invention with great flexibility for linkages to
scaffolds, macromolecules and other moieties. In particular, the
compounds of the invention may be reliably, securely and
efficiently covalently linked to scaffolds, such as an antibody.
Surprisingly, it has been found that locating the linking residue
at certain key positions in the peptide leads to increased
stability and/or binding of the peptide.
[0045] The linking residue is an amino acid residue available for
covalent bonding through the amino terminus, the carboxy terminus,
or the side chain of the linking residue. The linking residue may
be K. In other embodiments, the linking residue may be Y. In other
embodiments, the linking residue may be T. The linking residue may
be Dab. The linking residue may be Dap. In some embodiments, the
linking residue is selected from the group consisting of K, Dab,
Dap, Y, and T.
[0046] In other embodiments, the linking residue may be C. The
linking residue may be R. The linking residue may be S. The linking
residue may be N. The linking residue may be Q. The linking residue
may be D. The linking residue may be E.
[0047] The linking residue may be any one residue. In some
embodiments, the linking residue may be x.sup.9. The linking
residue may be x.sup.11. In some embodiments, using the side chain
of lysine and modified lysine residues as the linking residue has
been found to provide certain advantages, including permitting
specific, reliable, directional and efficient chemical covalent
linkages at that location.
[0048] Referring to the foregoing formulae (SEQ ID NOs: 161-178,
and SEQ ID NO: 193) and more generally:
x.sup.1 may be Q. x.sup.1 may be T. x.sup.1 may be S. x.sup.1 may
be R. x.sup.1 may be H. x.sup.2 may be N. x.sup.2 may be R. x.sup.2
may be H. x.sup.2 may be AcK. x.sup.2 may be Nick. x.sup.2 may be
CbcK. In some embodiments, x.sup.2 may be selected from the group
consisting of K, N, R, H, AcK, Nick, or CbcK. X.sup.2 may be
selected from the group consisting of N, R, H, AcK, Nick, or CbcK.
X.sup.2 may be selected from the group consisting of R, H, AcK,
Nick, or CbcK. X.sup.2 may be selected from the group consisting of
R, H, AcK, or CbcK. X.sup.2 may be selected from the group
consisting of R, H, or AcK. In some embodiments of the invention
comprising linkers and antibodies, x.sup.2 may be K. x.sup.3 may be
F. x.sup.3 may be Y. x.sup.3 may be W. x.sup.3 may be BPA. x.sup.3
may be CF. x.sup.3 may be or NF. x.sup.4 may be Q. x.sup.4 may be
M. x.sup.4 may be E. x.sup.4 may be K. x.sup.6 may be M. x.sup.6
may be L. x.sup.6 may be I. x.sup.8 may be E. x.sup.8 may be D.
X.sup.9 may be L. x.sup.9 may be I. X.sup.9 may be TA. X.sup.9 may
be ThA. X.sup.9 may be K. X.sup.9 may be AcK. X.sup.9 may be a
linking residue. x.sup.10 may be E. x.sup.10 may be D. x.sup.11 may
be Q. X.sup.11 may be N. X.sup.11 may be C. X.sup.11 may be K.
X.sup.11 may be AcK. X.sup.11 may be Dab. X.sup.11 may be Dap.
X.sup.11 may be a linking residue. x.sup.12 may be T. x.sup.12 may
be R. x.sup.12 may be K. x.sup.13 may be L. x.sup.13 may be HL.
x.sup.13 may be Nva. x.sup.13 may be I. x.sup.13 may be HchA.
x.sup.13 may be HF. x.sup.13 may be ThA. In some embodiments,
x.sup.13 may be selected from the group consisting of L, HL, Nva,
I, HchA, HF, and ThA. In some embodiments, x.sup.13 may be selected
from the group consisting of L, HL, and Nva. x.sup.14 may be F.
x.sup.14 may be Y. x.sup.14 may be W. x.sup.14 may be BPA. X.sup.14
may be CF. x.sup.14 may be or NF. x.sup.15 may be D. x.sup.15 may
be E. x.sup.16 may be Q. x.sup.16 may be N. x.sup.17 may be F.
x.sup.17 may be Y. x.sup.17 may be W. x.sup.17 may be BPA. X.sup.17
may be CF. x.sup.17 may be or NF. x.sup.18 may be M. x.sup.19 may
be L. x.sup.19 may be I. x.sup.20 may be Q. x.sup.20 may be N.
x.sup.21 may be Q. x.sup.21 may be N. x.sup.22 may be G.
[0049] In some aspects of the invention, AA targeting agents of the
invention may comprise any Angiopoietin 2 antagonist.
[0050] In some aspects of the invention, AA targeting agents of the
invention may comprise a peptide comprising a sequence
substantially homologous to one or more sequences selected from the
group consisting of: SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ
ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28,
SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID
NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ
ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42,
SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID
NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ
ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56,
SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID
NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ
ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70,
SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID
NO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ
ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84,
SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID
NO:89, SEQ ID NO:90, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ
ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98,
SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID
NO:103, SEQ ID NO:104, SEQ ID NO:105, SEQ ID NO:106, SEQ ID NO:107,
SEQ ID NO:108, SEQ ID NO:109, SEQ ID NO:110, SEQ ID NO:111, SEQ ID
NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:115, SEQ ID NO:116,
SEQ ID NO:117, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID
NO:121, SEQ ID NO:122, SEQ ID NO:123, SEQ ID NO:124, SEQ ID NO:125,
SEQ ID NO:126, SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, SEQ ID
NO:130, SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:133, SEQ ID NO:134,
SEQ ID NO:135, SEQ ID NO:136, SEQ ID NO:137, SEQ ID NO:138, SEQ ID
NO:139, SEQ ID NO:140, SEQ ID NO:141, SEQ ID NO:142, SEQ ID NO:143,
SEQ ID NO:144, SEQ ID NO:145, SEQ ID NO:146, SEQ ID NO:147, SEQ ID
NO:148, SEQ ID NO:149, SEQ ID NO:150, SEQ ID NO:151, SEQ ID NO:152,
SEQ ID NO:153, SEQ ID NO:154, SEQ ID NO:155, SEQ ID NO:156, SEQ ID
NO:157, SEQ ID NO:158, SEQ ID NO:159, SEQ ID NO:160, SEQ ID NO:161,
SEQ ID NO:162, SEQ ID NO:163, SEQ ID NO:164, SEQ ID NO:165, SEQ ID
NO:166, SEQ ID NO:167, SEQ ID NO:168, SEQ ID NO:169, SEQ ID NO:170,
SEQ ID NO:171, SEQ ID NO:172, SEQ ID NO:173, SEQ ID NO:174, SEQ ID
NO:175, SEQ ID NO:176, SEQ ID NO:177, SEQ ID NO:178, SEQ ID NO:191,
SEQ ID NO:192, and SEQ ID NO: 193.
[0051] In some aspects, the invention provides an AA targeting
agent selected from the group consisting of: [0052] R.sup.1-Q(AcK)Y
QPL DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:29) [0053]
R.sup.1-QNY QPL DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:28)
[0054] R.sup.1-Q(AcK)Y QPL DEK D(AcK)T LYD QFM LQQ G-R.sup.2; (SEQ
ID NO:30) [0055] R.sup.1-Q(AcK)Y QPL DEK DET LYD QFM LQQ G-R.sup.2;
(SEQ ID NO:33) [0056] R.sup.1-Q(AcK)Y Q(DHP)L DEL DKT LYD QFM LQQ
G-R.sup.2; (SEQ ID NO:34) [0057] R.sup.1-Q(AcK)Y Q(HP)L DEL DKT LYD
QFM LQQ G-R.sup.2; (SEQ ID NO:35) [0058] R.sup.1-Q(AcK)Y QPL DEL
DKT IYD QFM LQQ G-R.sup.2; (SEQ ID NO:36) [0059] R.sup.1-Q(AcK)Y
QPL DEI DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:37) [0060]
R.sup.1-Q(AcK)Y QPL DEL DKT (HChA)YD QFM LQQ G-R.sup.2; (SEQ ID
NO:38) [0061] R.sup.1-Q(AcK)Y QPL DEL DKT (HF)YD QFM LQQ G-R.sup.2;
(SEQ ID NO:39) [0062] R.sup.1-Q(AcK)Y QPL DEL DKT (ThA)YD QFM LQQ
G-R.sup.2; (SEQ ID NO:40) [0063] R.sup.1-Q(AcK)Y QPL DEL DKT
(Nva)YD QFM LQQ G-R.sup.2; (SEQ ID NO:41) [0064] R.sup.1-Q(AcK)Y
QPL DEL DKT (HL)YD QFM LQQ G-R.sup.2; (SEQ ID NO:42) [0065]
R.sup.1-Q(AcK)Y QPL DE(AcK) DKT LYD QFM LQQ G-R.sup.2; (SEQ ID
NO:43) [0066] R.sup.1-Q(AcK)Y QPL DEL DKT LFD QFM LQQ G-R.sup.2;
(SEQ ID NO:44) [0067] R.sup.1-Q(AcK)Y Q(HP)L DE(ThA) DKT L(NO2F)D
QFM LQQ G-R.sup.2; (SEQ ID NO:45) [0068] R.sup.1-Q(AcK)Y Q(HP)L
DE(ThA) DKT L(BPA)D QFM LQQ G-R.sup.2; (SEQ ID NO:46) [0069]
R.sup.1-Q(AcK)Y Q(HP)L DE(ThA) DKT L(CO2H)FD QFM LQQ G-R.sup.2;
(SEQ ID NO:47) [0070] R.sup.1-Q(AcK)Y QPL DE(AcK) DKT L(NO2F)D QFM
LQQ G-R.sup.2; (SEQ ID NO:48) [0071] R.sup.1-Q(AcK)Y QPL DE(AcK)
DKT LFD QFM LQQ G-R.sup.2 (SEQ ID NO:63) [0072] DCB-Q(AcK)Y QPL DEL
DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:49) [0073] DFB-Q(AcK)Y QPL
DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:50) [0074] PyC-Q(AcK)Y
QPL DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:51) [0075] Amido
2-PEG-Q(AcK)Y QPL DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:52)
[0076] R.sup.1-Q(CIBnCarbamateK)Y QPL DEL DKT LYD QFM LQQ
G-R.sup.2; (SEQ ID NO:53) [0077] R.sup.1-Q(AcK)Y QPL DEL D(Dab)T
LYD QFM LQQ G-R.sup.2; (SEQ ID NO:54) [0078] R.sup.1-Q(AcK)Y QPL
DEL D(Dap)T LYD QFM LQQ G-R.sup.2; (SEQ ID NO:55) [0079]
R.sup.1-Q(AcK)Y QPL DEL DKT L(NO2F)D QFM LQQ G-R.sup.2; (SEQ ID
NO:91) [0080] R.sup.1-QNY QPL DEL DKT L(BPA)D QFM LQQ G-R.sup.2;
(SEQ ID NO:56) [0081] R.sup.1-QNY QPL DEL DKT L(CF)D QFM LQQ
G-R.sup.2; (SEQ ID NO:57) [0082] R.sup.1-Q(Nick)Y QPL DEL DKT LYD
QFM LQQ G-R.sup.2; (SEQ ID NO:61) [0083] R.sup.1-Q(AcK)Y QPL
DE(AcK) DKT L(CF)D QFM LQQ G-R.sup.2; (SEQ ID NO:62) and [0084]
R.sup.1-Q(AcK)Y QPL DE(AcK) DKT LFD QFM LQQ G-R.sup.2; (SEQ ID NO:
63) [0085] wherein [0086] R.sup.1 is CH.sub.3, C(O)CH.sub.3,
C(O)CH.sub.3, C(O)CH.sub.2CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.2CH.sub.3, C(O)C.sub.6H.sub.5,
C(O)CH.sub.2CH.sub.2(CH.sub.2CH.sub.2O).sub.1-5Me, dichlorobenzoyl
(DCB), difluorobenzoyl (DFB), pyridinyl carboxlate (PyC) or
amido-2-PEG, an amino protecting group, a lipid fatty acid group or
a carbohydrate; and [0087] R.sup.2 is OH, NH.sub.2, NH(CH.sub.3),
NHCH.sub.2CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.2CH.sub.3, NHC.sub.6H.sub.5,
NHCH.sub.2CH.sub.2OCH.sub.3, NHOCH.sub.3, NHOCH.sub.2CH.sub.3, a
carboxy protecting group, a lipid fatty acid group or a
carbohydrate.
[0088] In some aspects, the invention provides an AA targeting
agent-linker conjugate having Formula I:
Linker-[AA targeting agent] (I) [0089] wherein: [0090] [AA
targeting agent] is a peptide selected from the group consisting
of: [0091] R.sup.1-QKY QPL DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID
NO:21) [0092] R.sup.1-Q(AcK)Y QPL DEL DKT LYD QFM LQQ G-R.sup.2;
(SEQ ID NO:22) [0093] R.sup.1-QNY QPL DEL DKT LYD QFM LQQ
G-R.sup.2; (SEQ ID NO:23) [0094] R.sup.1-Q(AcK)Y QPL DEK D(AcK)T
LYD QFM LQQ G-R.sup.2; (SEQ ID NO:30) [0095] R.sup.1-Q(AcK)Y QPL
DEK DET LYD QFM LQQ G-R.sup.2; (SEQ ID NO:33) [0096]
R.sup.1-Q(AcK)Y Q(DHP)L DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID
NO:34) [0097] R.sup.1-Q(AcK)Y Q(HP)L DEL DKT LYD QFM LQQ G-R.sup.2;
(SEQ ID NO:35) [0098] R.sup.1-Q(AcK)Y QPL DEL DKT IYD QFM LQQ
G-R.sup.2; (SEQ ID NO:36) [0099] R.sup.1-Q(AcK)Y QPL DEI DKT LYD
QFM LQQ G-R.sup.2; (SEQ ID NO:37) [0100] R.sup.1-Q(AcK)Y QPL DEL
DKT (HChA)YD QFM LQQ G-R.sup.2; (SEQ ID NO:38) [0101]
R.sup.1-Q(AcK)Y QPL DEL DKT (HF)YD QFM LQQ G-R.sup.2; (SEQ ID
NO:39) [0102] R.sup.1-Q(AcK)Y QPL DEL DKT (ThA)YD QFM LQQ
G-R.sup.2; (SEQ ID NO:40) [0103] R.sup.1-Q(AcK)Y QPL DEL DKT
(Nva)YD QFM LQQ G-R.sup.2; (SEQ ID NO:41) [0104] R.sup.1-Q(AcK)Y
QPL DEL DKT (HL)YD QFM LQQ G-R.sup.2; (SEQ ID NO:42) [0105]
R.sup.1-Q(AcK)Y QPL DE(AcK) DKT LYD QFM LQQ G-R.sup.2; (SEQ ID
NO:43) [0106] R.sup.1-Q(AcK)Y QPL DEL DKT LFD QFM LQQ G-R.sup.2;
(SEQ ID NO:44) [0107] R.sup.1-Q(AcK)Y Q(HP)L DE(ThA) DKT L(NO2F)D
QFM LQQ G-R.sup.2; (SEQ ID NO:45) [0108] R.sup.1-Q(AcK)Y Q(HP)L
DE(ThA) DKT L(BPA)D QFM LQQ G-R.sup.2; (SEQ ID NO:46) [0109]
R.sup.1-Q(AcK)Y Q(HP)L DE(ThA) DKT L(CO2H)FD QFM LQQ G-R.sup.2;
(SEQ ID NO:47) [0110] R.sup.1-Q(AcK)Y QPL DE(AcK) DKT L(NO2F)D QFM
LQQ G-R.sup.2; (SEQ ID NO:48) [0111] R.sup.1-Q(AcK)Y QPL DE(AcK)
DKT LFD QFM LQQ G-R.sup.2; (SEQ ID NO:68) [0112] DCB-Q(AcK)Y QPL
DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:49) [0113] DFB-Q(AcK)Y
QPL DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:50) [0114]
PyC-Q(AcK)Y QPL DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:51)
[0115] Amido 2-PEG-Q(AcK)Y QPL DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ
ID NO:52) [0116] R.sup.1-Q(CIBnCarbamateK)Y QPL DEL DKT LYD QFM LQQ
G-R.sup.2; (SEQ ID NO:53) [0117] R.sup.1-Q(AcK)Y QPL DEL D(Dab)T
LYD QFM LQQ G-R.sup.2; (SEQ ID NO:54) [0118] R.sup.1-Q(AcK)Y QPL
DEL D(Dap)T LYD QFM LQQ G-R.sup.2; (SEQ ID NO:55) [0119]
R.sup.1-Q(AcK)Y QPL DEL DKT L(NO2F)D QFM LQQ G-R.sup.2; (SEQ ID
NO:91) [0120] R.sup.1-QNY QPL DEL DKT L(BPA)D QFM LQQ G-R.sup.2;
(SEQ ID NO:56) [0121] R.sup.1-QNY QPL DEL DKT L(CF)D QFM LQQ
G-R.sup.2; (SEQ ID NO:57) [0122] R.sup.1-QNY QPL DEL DKT LYD QFM
LQQ G-R.sup.2; (SEQ ID NO:58) [0123] R.sup.1-Q(Nick)Y QPL DEL DKT
LYD QFM LQQ G-R.sup.2; (SEQ ID NO:61) [0124] R.sup.1-Q(AcK)Y QPL
DE(AcK) DKT L(CF)D QFM LQQ G-R.sup.2; (SEQ ID NO:62) and [0125]
R.sup.1-Q(AcK)Y QPL DE(AcK) DKT LFD QFM LQQ G-R.sup.2; (SEQ ID
NO:63) wherein [0126] R.sup.1 refers to the amino group portion of
the amino terminus residue of a targeting agent and is NH.sub.2,
NHC(O)CH.sub.3, NHC(O)CH.sub.2CH.sub.3,
NHC(O)CH.sub.2CH.sub.2CH.sub.3, NHC(O)CH(CH.sub.3)CH.sub.3,
NHC(O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
NHC(O)CH(CH.sub.3)CH.sub.2CH.sub.3, NHC(O)C.sub.6H.sub.5,
NH(CH.sub.3)C(O)CH.sub.2CH.sub.2(CH.sub.2CH.sub.2O).sub.1-5Me, an
amino protecting group, a lipid fatty acid group or a carbohydrate;
[0127] R.sup.2 refers to the carboxy terminus portion of the
carboxy terminus residue of a targeting agent and is COOH,
C(O)NH.sub.2, C(O)NH(CH.sub.3), C(O)NHCH.sub.2CH.sub.3,
C(O)NHCH.sub.2CH.sub.2CH.sub.3, C(O)NHCH(CH.sub.3)CH.sub.3,
C(O)NHCH.sub.2CH.sub.2CH.sub.2CH.sub.3,
C(O)NHCH(CH.sub.3)CH.sub.2CH.sub.3, C(O)NHC.sub.6H.sub.5,
C(O)NHCH.sub.2CH.sub.2OCH.sub.3, C(O)NHOCH.sub.3,
C(O)NHOCH.sub.2CH.sub.3, a carboxy protecting group, a lipid fatty
acid group or a carbohydrate; and [0128] Linker is a linker moiety
having the formula --X-Recognition GroupY--Z, wherein: [0129] X is
optionally present, and is a biologically compatible polymer, block
copolymer C, H, N, O, P, S, halogen (F, Cl, Br, I), or a salt
thereof, alkyl, alkenyl, alkynyl, oxoalkyl, oxoalkenyl, oxoalkynyl,
aminoalkyl, aminoalkenyl, aminoalkynyl, sulfoalkyl, sulfoalkenyl,
sulfoalkynyl, phosphoalkyl, phosphoalkenyl, or phosphoalkynyl
group, attached to one of the residues that comprises an AA
targeting agent; [0130] Recognition GroupY is an optionally present
recognition group comprising at least a ring structure; and [0131]
Z is a reactive group that is capable of covalently linking to a
side chain in a combining site of an antibody; and pharmaceutically
acceptable salts, stereoisomers, tautomers, solvates, and prodrugs
thereof.
[0132] In describing aspects of the invention as above, the initial
NH group or R.sup.1 is provided by the N-terminal amino-acid
residue of the peptide in question: thus, NHC(O)CH.sub.3 represents
a C(O)CH.sub.3 group covalently bonded to the N-terminus of the
peptide, and may elsewhere throughout this specification and claims
be written as C(O)CH.sub.3, and so on. Accordingly, R.sup.1 may be
written as: [0133] R.sup.1 is CH.sub.3, C(O)CH.sub.3, C(O)CH.sub.3,
C(O)CH.sub.2CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.2CH.sub.3, C(O)C.sub.6H.sub.5,
C(O)CH.sub.2CH.sub.2(CH.sub.2CH.sub.2O).sub.1-5Me, dichlorobenzoyl
(DCB), difluorobenzoyl (DFB), pyridinyl carboxlate (PyC) or
amido-2-PEG, an amino protecting group, a lipid fatty acid group or
a carbohydrate; and
[0134] Similarly, in describing aspects of the invention as above,
the initial C(O) group of R.sup.2 is provided by the C-terminal
amino-acid residue of the peptide in question: thus, C(O)NH.sub.2
represents a NH.sub.2 group covalently bonded to the C-terminus of
the peptide, and may else where be written as NH.sub.2, and so on.
Accordingly, R.sup.2 may be written as: [0135] R.sup.2 is OH,
NH.sub.2, NH(CH.sub.3), NHCH.sub.2CH.sub.3,
NHCH.sub.2CH.sub.2CH.sub.3, NHCH(CH.sub.3)CH.sub.3,
NHCH.sub.2CH.sub.2CH.sub.2CH.sub.3, NHCH(CH.sub.3)CH.sub.2CH.sub.3,
NHC.sub.6H.sub.5, NHCH.sub.2CH.sub.2OCH.sub.3, NHOCH.sub.3,
NHOCH.sub.2CH.sub.3, a carboxy protecting group, a lipid fatty acid
group or a carbohydrate.
[0136] In the listed peptide sequences, non-standard amino acid
residues are enclosed within parentheses. In some cases, the amino
terminus of a peptide is attached to one of the following capping
groups: dichlorobenzoyl (DCB), difluorobenzoyl (DFB), pyridinyl
carboxlate (PyC) or amido-2-PEG.
[0137] In other aspects, the invention provides compounds according
to: [0138] A compound having the formula selected from the group
consisting of: [0139] R.sup.1-Q(AcK)Y QPL DEL DK(Linker)T LYD QFM
LQQ G-R.sup.2 (SEQ ID NO:132); [0140] R.sup.1-Q(AcK)Y QPL DEL
DK(Linker)T L(NO2F)D QFM LQQ G-R.sup.2; (SEQ ID NO:133) [0141]
R.sup.1-Q(AcK)Y QPL DEK(Linker) DK(AcK)T LYD QFM LQQ G-R.sup.2 (SEQ
ID NO:134); and [0142] R.sup.1-Q(AcK)Y QPL DEK(AcK) DK(Linker)T LYD
QFM LQQ G-R.sup.2 (SEQ ID NO:135); wherein [0143] R.sup.1 is
CH.sub.3, C(O)CH.sub.3, C(O)CH.sub.3, C(O)CH.sub.2CH.sub.3,
C(O)CH.sub.2CH.sub.2CH.sub.3, C(O)CH(CH.sub.3)CH.sub.3,
C(O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.2CH.sub.3, C(O)C.sub.6H.sub.5,
C(O)CH.sub.2CH.sub.2(CH.sub.2CH.sub.2O).sub.1-5Me, dichlorobenzoyl
(DCB), difluorobenzoyl (DFB), pyridinyl carboxlate (PyC) or
amido-2-PEG, an amino protecting group, a lipid fatty acid group or
a carbohydrate; and [0144] R.sup.2 is OH, NH.sub.2, NH(CH.sub.3),
NHCH.sub.2CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.2CH.sub.3, NHC.sub.6H.sub.5,
NHCH.sub.2CH.sub.2OCH.sub.3, NHOCH.sub.3, NHOCH.sub.2CH.sub.3, a
carboxy protecting group, a lipid fatty acid group or a
carbohydrate, and K(Linker) is a lysine reside attached to a linker
(Linker) wherein (Linker) is capable of forming a covalent bond
with an amino acid side chain in a combining site of an
antibody.
[0145] In some embodiments, K(Linker) is
##STR00001## [0146] wherein Linker is a linker moiety having the
formula --X-Recognition GroupY--Z--, wherein: [0147] X is:
[0147] ##STR00002## [0148] wherein v and w are selected such that
the backbone length of X is 6-12 atoms; [0149] Recognition GroupY
is a recognition group comprising at least a ring structure; and
[0150] Z is a reactive group that is capable of forming a covalent
bond with an amino acid side chain in a combining site of an
aldolase antibody.
[0151] In some embodiments Recognition GroupY has the optionally
substituted structure:
##STR00003##
wherein a, b, c, d, and e are independently carbon or nitrogen; f
is carbon, nitrogen, oxygen, or sulfur; Recognition Group Y is
attached to X and Z independently at any two ring positions of
sufficient valence; and no more than four of a, b, c, d, e, or f
are simultaneously nitrogen.
[0152] In some embodiments, Z is selected from the group consisting
of substituted 1,3-diketones or acyl beta-lactams.
[0153] In some embodiments Z has the structure:
##STR00004##
wherein q=0, 1, 2, 3, 4, or 5. In other embodiments, q=1, 2, or
3.
[0154] In some embodiments of compounds of Formula I, X is:
--R.sup.22--P--R.sup.23-- or
--R.sup.22--P--R.sup.21--P'--R.sup.23--
wherein: [0155] P and P' are independently selected from the group
consisting of polyoxyalkylene oxides such as polyethylene oxide,
polyethyloxazoline, poly-N-vinyl pyrrolidone, polyvinyl alcohol,
polyhydroxyethyl acrylate, polyhydroxy ethylmethacrylate and
polyacrylamide, polyamines having amine groups on either the
polymer backbone or the polymer sidechains, such as polylysine,
polyornithine, polyarginine, and polyhistidine, nonpeptide
polyamines such as polyaminostyrene, polyaminoacrylate,
poly(N-methyl aminoacrylate), poly(N-ethylaminoacrylate),
poly(N,N-dimethyl aminoacrylate), poly(N,N-diethylaminoacrylate),
poly(aminomethacrylate), poly(N-methyl aminomethacrylate),
poly(N-ethyl aminomethacrylate), poly(N,N-dimethyl
aminomethacrylate), poly(N,N-diethyl aminomethacrylate),
poly(ethyleneimine), polymers of quaternary amines, such as
poly(N,N,N-trimethylaminoacrylate chloride),
poly(methyacrylamidopropyltrimethyl ammonium chloride),
proteoglycans such as chondroitin sulfate-A (4-sulfate) chondroitin
sulfate-C(6-sulfate) and chondroitin sulfate-B, polypeptides such
as polyserine, polythreonine, polyglutamine, natural or synthetic
polysaccharides such as chitosan, hydroxy ethyl cellulose, and
lipids; [0156] R.sup.21, R.sup.22, and R.sup.23 are each
independently a covalent bond, --O--, --S--, --NR.sup.b--, amide,
substituted or unsubstituted straight or branched chain C.sub.1-50
alkylene, or substituted or unsubstituted straight or branched
chain C.sub.1-50 heteroalkylene; [0157] R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.m alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl; and [0158] R.sup.21, R.sup.22,
and R.sup.23 are selected such that the backbone length of X
remains about 200 atoms or less.
[0159] In some embodiments of compounds of Formula I, X is attached
to an amino acid residue in [AA targeting agent], and is an
optionally substituted
--R.sup.22--[CH.sub.2--CH.sub.2--O].sub.t--R.sup.23--,
--R.sup.22-cycloalkyl-R.sup.23--, --R.sup.22-aryl-R.sup.23--, or
--R.sup.22-heterocyclyl-R.sup.23--, wherein t is 0 to 50.
[0160] In some embodiments of compounds of Formula I, R.sup.22 is
--(CH.sub.2).sub.v--, --(CH.sub.2).sub.u--C(O)--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(O)--O--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(S)--NR.sup.b--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(O)--NR.sup.b--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--NR.sup.b--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--O--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--S(O).sub.0-2--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--S(O).sub.0-2--NR.sup.b--(CH.sub.2).sub.v--, or
--(CH.sub.2).sub.u--P(O)(OR.sup.b)--O--(CH.sub.2).sub.v--, wherein
u and v are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19 or 20.
[0161] In some embodiments of compounds of Formula I, R.sup.21 and
R.sup.23 are independently --(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--C(O)--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--C(O)--O--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.r--C(S)--NR.sup.b--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--C(O)--NR.sup.b--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--NR.sup.b--(CH.sub.2).sub.s--,
--(CH.sub.2)--O--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--S(O).sub.0-2--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--S(O).sub.0-2--NR.sup.b--(CH.sub.2).sub.s--, or
--(CH.sub.2).sub.r--P(O)(OR.sup.b)--O--(CH.sub.2).sub.s--, wherein
r, s, and v are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,
12, 13, 14, 15, 16, 17, 18, 19 or 20.
[0162] In some embodiments of Formula I, if t>1 or if X is
--R.sup.22--[CH.sub.2--CH.sub.2--O].sub.t--R.sup.23--,
--R.sup.22-cycloalkyl-R.sup.23, --R.sup.22-aryl-R.sup.23--, or
--R.sup.22-heterocyclyl-R.sup.23--, Y is present.
[0163] Exemplary compounds in accordance with Formula I, wherein
R.sup.1 is either NHC(O)CH.sub.3 or DFB, R.sup.2 is C(O)NH.sub.2,
and the X portion of the linker moiety is "O" PEG, include:
##STR00005## ##STR00006##
[0164] Another aspect of the invention, illustrated in Formula II,
is an AA targeting compound comprising an AA targeting agent
covalently linked to a combining site of an Antibody via an
intervening linker Linker'. The Antibody portion of an AA targeting
compound can include whole (full length) antibody, unique antibody
fragments, or any other forms of an antibody as this term is used
herein. In one embodiment, the Antibody is a humanized version of a
murine aldolase antibody comprising a constant region from a human
IgG, IgA, IgM, IgD, or IgE antibody. In another embodiment, the
Antibody is a chimeric antibody comprising the variable region from
a murine aldolase antibody and a constant region from a human IgG,
IgA, IgM, IgD, or IgE antibody. In a further embodiment, the
Antibody is a fully human version of a murine aldolase antibody
comprising a polypeptide sequence from natural or native human IgG,
IgA, IgM, IgD, or IgE antibody
Antibody-Linker'-[AA targeting agent] (II)
wherein: [AA targeting agent] is a peptide selected from the group
consisting of: [0165] R.sup.1-QKY QPL DEL DKT LYD QFM LQQ
G-R.sup.2; (SEQ ID NO:21) [0166] R.sup.1-Q(AcK)Y QPL DEL DKT LYD
QFM LQQ G-R.sup.2; (SEQ ID NO:22) [0167] R.sup.1-QNY QPL DEL DKT
LYD QFM LQQ G-R.sup.2; (SEQ ID NO:23) [0168] R.sup.1-Q(AcK)Y QPL
DEK D(AcK)T LYD QFM LQQ G-R.sup.2; (SEQ ID NO:30) [0169]
R.sup.1-Q(AcK)Y QPL DEK DET LYD QFM LQQ G-R.sup.2; (SEQ ID NO:33)
[0170] R.sup.1-Q(AcK)Y Q(DHP)L DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ
ID NO:34) [0171] R.sup.1-Q(AcK)Y Q(HP)L DEL DKT LYD QFM LQQ
G-R.sup.2; (SEQ ID NO:35) [0172] R.sup.1-Q(AcK)Y QPL DEL DKT IYD
QFM LQQ G-R.sup.2; (SEQ ID NO:36) [0173] R.sup.1-Q(AcK)Y QPL DEI
DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:37) [0174] R.sup.1-Q(AcK)Y
QPL DEL DKT (HChA)YD QFM LQQ G-R.sup.2; (SEQ ID NO:38) [0175]
R.sup.1-Q(AcK)Y QPL DEL DKT (HF)YD QFM LQQ G-R.sup.2; (SEQ ID
NO:39) [0176] R.sup.1-Q(AcK)Y QPL DEL DKT (ThA)YD QFM LQQ
G-R.sup.2; (SEQ ID NO:40) [0177] R.sup.1-Q(AcK)Y QPL DEL DKT
(Nva)YD QFM LQQ G-R.sup.2; (SEQ ID NO:41) [0178] R.sup.1-Q(AcK)Y
QPL DEL DKT (HL)YD QFM LQQ G-R.sup.2; (SEQ ID NO:42) [0179]
R.sup.1-Q(AcK)Y QPL DE(AcK) DKT LYD QFM LQQ G-R.sup.2; (SEQ ID
NO:43) [0180] R.sup.1-Q(AcK)Y QPL DEL DKT LFD QFM LQQ G-R.sup.2;
(SEQ ID NO:44) [0181] R.sup.1-Q(AcK)Y Q(HP)L DE(ThA) DKT L(NO2F)D
QFM LQQ G-R.sup.2; (SEQ ID NO:45) [0182] R.sup.1-Q(AcK)Y Q(HP)L
DE(ThA) DKT L(BPA)D QFM LQQ G-R.sup.2; (SEQ ID NO:46) [0183]
R.sup.1-Q(AcK)Y Q(HP)L DE(ThA) DKT L(CO2H)FD QFM LQQ G-R.sup.2;
(SEQ ID NO:47) [0184] R.sup.1-Q(AcK)Y QPL DE(AcK) DKT L(NO2F)D QFM
LQQ G-R.sup.2; (SEQ ID NO:48) [0185] R.sup.1-Q(AcK)Y QPL DE(AcK)
DKT LFD QFM LQQ G-R.sup.2; (SEQ ID NO:68) [0186] DCB-Q(AcK)Y QPL
DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:49) [0187] DFB-Q(AcK)Y
QPL DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:50) [0188]
PyC-Q(AcK)Y QPL DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:51)
[0189] Amido 2-PEG-Q(AcK)Y QPL DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ
ID NO:52) [0190] R.sup.1-Q(CIBnCarbamateK)Y QPL DEL DKT LYD QFM LQQ
G-R.sup.2; (SEQ ID NO:53) [0191] R.sup.1-Q(AcK)Y QPL DEL D(Dab)T
LYD QFM LQQ G-R.sup.2; (SEQ ID NO:54) [0192] R.sup.1-Q(AcK)Y QPL
DEL D(Dap)T LYD QFM LQQ G-R.sup.2; (SEQ ID NO:55) [0193]
R.sup.1-Q(AcK)Y QPL DEL DKT L(NO2F)D QFM LQQ G-R.sup.2; (SEQ ID
NO:91) [0194] R.sup.1-QNY QPL DEL DKT L(BPA)D QFM LQQ G-R.sup.2;
(SEQ ID NO:56) [0195] R.sup.1-QNY QPL DEL DKT L(CF)D QFM LQQ
G-R.sup.2; (SEQ ID NO:57) [0196] R.sup.1-QNY QPL DEL DKT LYD QFM
LQQ G-R.sup.2; (SEQ ID NO:58) [0197] R.sup.1-Q(Nick)Y QPL DEL DKT
LYD QFM LQQ G-R.sup.2; (SEQ ID NO:61) [0198] R.sup.1-Q(AcK)Y QPL
DE(AcK) DKT L(CF)D QFM LQQ G-R.sup.2; (SEQ ID NO:62) and [0199]
R.sup.1-Q(AcK)Y QPL DE(AcK) DKT LFD QFM LQQ G-R.sup.2; (SEQ ID
NO:63) wherein [0200] R.sup.1 is CH.sub.3, C(O)CH.sub.3,
C(O)CH.sub.3, C(O)CH.sub.2CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.2CH.sub.3, C(O)C.sub.6H.sub.5,
C(O)CH.sub.2CH.sub.2(CH.sub.2CH.sub.2O).sub.1-5Me, dichlorobenzoyl
(DCB), difluorobenzoyl (DFB), pyridinyl carboxlate (PyC) or
amido-2-PEG, an amino protecting group, a lipid fatty acid group or
a carbohydrate; and [0201] R.sup.2 is OH, NH.sub.2, NH(CH.sub.3),
NHCH.sub.2CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.2CH.sub.3, NHC.sub.6H.sub.5,
NHCH.sub.2CH.sub.2OCH.sub.3, NHOCH.sub.3, NHOCH.sub.2CH.sub.3, a
carboxy protecting group, a lipid fatty acid group or a
carbohydrate, and [0202] Linker' is a linker moiety having the
formula --X-Recognition Group Y--Z', wherein: [0203] X is a
biologically compatible polymer or block copolymer attached to one
of the residues that comprises an AA targeting agent; [0204]
Recognition Group Y is an optionally present recognition group
comprising at least a ring structure; and [0205] Z is a group that
is covalently linked to a side chain in a combining site of an
antibody; and pharmaceutically acceptable salts, stereoisomers,
tautomers, solvates, and prodrugs thereof.
[0206] In the listed peptide sequences, non-standard amino acid
residues are enclosed within parentheses. In some cases, the amino
terminus of a peptide is attached to one of the following capping
groups: dichlorobenzoyl (DCB), difluorobenzoyl (DFB), pyridinyl
carboxlate (PyC) or amido-2-PEG.
[0207] In some embodiments of compounds of Formula II, X is:
--R.sup.22--P--R.sup.23-- or
--R.sup.22--P--R.sup.21--P'--R.sup.23--
wherein: [0208] P and P' are independently selected from the group
consisting of polyoxyalkylene oxides such as polyethylene oxide,
polyethyloxazoline, poly-N-vinyl pyrrolidone, polyvinyl alcohol,
polyhydroxyethyl acrylate, polyhydroxy ethylmethacrylate and
polyacrylamide, polyamines having amine groups on either the
polymer backbone or the polymer side chains, such as polylysine,
polyornithine, polyarginine, and polyhistidine, nonpeptide
polyamines such as polyaminostyrene, polyaminoacrylate,
poly(N-methyl aminoacrylate), poly(N-ethylaminoacrylate),
poly(N,N-dimethyl aminoacrylate), poly(N,N-diethylaminoacrylate),
poly(aminomethacrylate), poly(N-methyl aminomethacrylate),
poly(N-ethyl aminomethacrylate), poly(N,N-dimethyl
aminomethacrylate), poly(N,N-diethyl aminomethacrylate),
poly(ethyleneimine), polymers of quaternary amines, such as
poly(N,N,N-trimethylaminoacrylate chloride),
poly(methyacrylamidopropyltrimethyl ammonium chloride),
proteoglycans such as chondroitin sulfate-A (4-sulfate) chondroitin
sulfate-C(6-sulfate) and chondroitin sulfate-B, polypeptides such
as polyserine, polythreonine, polyglutamine, natural or synthetic
polysaccharides such as chitosan, hydroxy ethyl cellulose, and
lipids; [0209] R.sup.21, R.sup.22, and R.sup.23 are each
independently a covalent bond, --O--, --S--, --NR.sup.b--,
substituted or unsubstituted straight or branched chain C.sub.1-50
alkylene, or substituted or unsubstituted straight or branched
chain C.sub.1-50 heteroalkylene; [0210] R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.m alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl; and [0211] R.sup.21, R.sup.22,
and R.sup.23 are selected such that the backbone length of X
remains about 200 atoms or less.
[0212] In some embodiments of compounds of Formula II, X is
attached to an amino acid residue in [AA targeting agent], and is
an optionally substituted
--R.sup.22--[CH.sub.2--CH.sub.2--O].sub.t--R.sup.23--,
--R.sup.22-cycloalkyl-R.sup.23--, --R.sup.22-aryl-R.sup.23--, or
--R.sup.22-heterocyclyl-R.sup.23--, wherein t is 0 to 50.
[0213] In some embodiments of compounds of Formula II, R.sup.22 is
--(CH.sub.2)--, --(CH.sub.2).sub.u--C(O)--(CH.sub.2)--,
--(CH.sub.2).sub.u--C(O)--O--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(S)--NR.sup.b--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(O)--NR.sup.b--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--NR.sup.b--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--O--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--S(O).sub.0-2--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--S(O).sub.0-2--NR.sup.b--(CH.sub.2).sub.v--, or
--(CH.sub.2).sub.u--P(O)(OR.sup.b)--O--(CH.sub.2).sub.v--, wherein
u and v are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19 or 20.
[0214] In some embodiments of compounds of Formula II, R.sup.21 and
R.sup.23 are independently --(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--C(O)--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--C(O)--O--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.r--C(S)--NR.sup.b--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--C(O)--NR.sup.b--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--NR.sup.b--(CH.sub.2).sub.s--,
--(CH.sub.2)--O--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--S(O).sub.0-2--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--S(O).sub.0-2--NR.sup.b--(CH.sub.2).sub.s--, or
--(CH.sub.2).sub.r--P(O)(OR.sup.b)--O--(CH.sub.2).sub.s--, wherein
r, s, and v are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,
12, 13, 14, 15, 16, 17, 18, 19 or 20.
[0215] Some aspects of the invention provide a compound selected
from the groups consisting of: [0216] R.sup.1-Q(AcK)Y QPL DEL
DK(Linker')T LYD QFM LQQ G-R.sup.2; (SEQ ID NO: 136) [0217]
R.sup.1-Q(AcK)Y QPL DEL DK(Linker')T L(NO2F)D QFM LQQ G-R.sup.2;
(SEQ ID NO: 137) [0218] R.sup.1-Q(AcK)Y QPL DEK(Linker') DK(Ac)T
LYD QFM LQQ G-R.sup.2; (SEQ ID NO: 138) and [0219] R.sup.1-Q(AcK)Y
QPL DE(AcK) DK(Linker')T LYD QFM LQQ G-R.sup.2; (SEQ ID NO: 139)
wherein [0220] R.sup.1 is CH.sub.3, C(O)CH.sub.3, C(O)CH.sub.3,
C(O)CH.sub.2CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.2CH.sub.3, C(O)C.sub.6H.sub.5,
C(O)CH.sub.2CH.sub.2(CH.sub.2CH.sub.2O).sub.1-5Me, dichlorobenzoyl
(DCB), difluorobenzoyl (DFB), pyridinyl carboxlate (PyC) or
amido-2-PEG, an amino protecting group, a lipid fatty acid group or
a carbohydrate; and [0221] R.sup.2 is OH, NH.sub.2, NH(CH.sub.3),
NHCH.sub.2CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.2CH.sub.3, NHC.sub.6H.sub.5,
NHCH.sub.2CH.sub.2OCH.sub.3, NHOCH.sub.3, NHOCH.sub.2CH.sub.3, a
carboxy protecting group, a lipid fatty acid group or a
carbohydrate, and K(Linker') is a lysine reside attached to a
linker Linker' wherein Linker' is covalent bonded with an amino
acid side chain in a combining site of an antibody.
[0222] In some embodiments K(Linker') is
##STR00007## [0223] Linker' is a linker moiety having the formula
--X-Recognition Group Y--Z'--, wherein: [0224] X is:
[0224] ##STR00008## [0225] wherein v and w are selected such that
the backbone length of X is 6-12 atoms; [0226] Recognition Group Y
is a recognition group comprising at least a ring structure; and
[0227] Z' is an attachment moiety comprising a covalent link to an
amino acid side chain in a combining site of an antibody.
[0228] In some embodiments Recognition GroupY has the optionally
substituted structure:
##STR00009##
wherein a, b, c, d, and e are independently carbon or nitrogen; f
is carbon, nitrogen, oxygen, or sulfur; Recognition Group Y is
attached to X and Z' independently at any two ring positions of
sufficient valence; and no more than four of a, b, c, d, e, or f
are simultaneously nitrogen.
[0229] In some embodiments Z' has the structure:
##STR00010##
Wherein q=0, 1, 2, 3, 4, or 5 and --N-Antibody refers to a covalent
link to an amino acid side chain in a combining site of an antibody
bearing an amino group. In other aspects, q=1, 2 or 3.
[0230] Another aspect of the invention, illustrated in Formula III,
is an AA targeting compound in which two AA targeting agents, which
may be the same or different, are each covalently linked to a
combining site of an antibody. The Antibody portion of an AA
targeting compound can include whole (full length) antibody, unique
antibody fragments, or any other forms of an antibody as this term
is used herein. In one embodiment, the Antibody is a humanized
version of a murine aldolase antibody comprising a constant region
from a human IgG, IgA, IgM, IgD, or IgE antibody. In another
embodiment, the Antibody is a chimeric antibody comprising the
variable region from a murine aldolase antibody and a constant
region from a human IgG, IgA, IgM, IgD, or IgE antibody. In a
further embodiment, the Antibody is a fully human version of a
murine aldolase antibody comprising a polypeptide sequence from
natural or native human IgG, IgA, IgM, IgD, or IgE antibody.
Antibody[-Linker'-[AA targeting agent]].sub.2 (III) [0231] wherein:
[0232] [AA targeting agent], Antibody, and Linker' are as defined
according to Formula II.
[0233] Also provided are methods of delivering or administering AA
targeting compounds of the invention and methods of treatment using
AA targeting compounds of the invention. For example, methods of
treating (including preventing) a disease or condition associated
with abnormal angiogenesis in a subject include administering a
therapeutically effective amount of an AA targeting compound of the
invention to the subject. Diseases and conditions that may be
treated include cancer, arthritis, hypertension, kidney disease,
psoriasis, angiogenesis of the eye associated with ocular disorder,
infection or surgical intervention, macular degeneration, diabetic
retinopathy, and the like.
[0234] Another aspect of the invention includes methods of using AA
targeting compounds of the invention for diagnostic purposes. For
example, the AA targeting compounds can be used for the diagnosis
of a disease or condition associated with abnormal angiogenesis,
including cancer, arthritis, psoriasis, angiogenesis of the eye
associated with an ocular disorder, infection or surgical
intervention, macular degeneration, diabetic retinopathy, and the
like.
BRIEF DESCRIPTION OF THE DRAWINGS
[0235] FIG. 1 illustrates one embodiment according to Formula II or
Formula III.
[0236] FIG. 2 illustrates one embodiment according to Formula II or
Formula III.
[0237] FIG. 3 illustrates one embodiment according to Formula II or
Formula III.
[0238] FIGS. 4A and 4b each illustrate one embodiment according to
Formula II or Formula III.
[0239] FIGS. 5A and 5b each illustrate one embodiment according to
Formula II or Formula III.
[0240] FIG. 6A and FIG. 6B illustrate the solid phase synthesis of
targeting agent-linker conjugates of the present invention.
[0241] FIG. 7A illustrates the amino acid sequence alignment of the
variable domains of m38c2, h38c2, and human germlines. Framework
regions (FR) and complementarity determining regions (CDR) are
defined according to Kabat et al. Asterisks mark differences
between m38c2 and h38c2 or between h38c2 and the human
germlines.
[0242] FIG. 7B illustrates the amino acid sequence of the light and
heavy chains (SEQ ID NOs:189 and 190, respectively) of one
embodiment of a humanized 38c2 IgG1.
[0243] FIG. 8 shows various structures that may serve as linker
reactive groups. Structures A-C form reversible covalent bonds with
surface accessible reactive nucleophilic groups (e.g., lysine or
cysteine side chain) of a combining site of an antibody. R'.sub.1,
R'.sub.2, R'.sub.3, and R.sub.4 in structures A-C represent
substituents which include, for example, C, H, N, O, P, S, halogen
(F, Cl, Br, I) or a salt thereof. X is N, C, or any other
heteroatom. These substituents may also include a group such as an
alkyl, alkenyl, alkynyl, oxoalkyl, oxoalkenyl, oxoalkynyl,
aminoalkyl, aminoalkenyl, aminoalkynyl, sulfoalkyl, sulfoalkenyl,
or sulfoalkynyl group, phosphoalkyl, phosphoalkenyl, phosphoalkynyl
group. R'.sub.2 and R'.sub.3 could be cyclic as exemplified in
structures B and C while X could be a heteroatom. For example,
structure A could form an irreversible covalent bond with a
reactive nucleophile if X is N and if R'.sub.1 and R.sub.3 form
part of a cyclic structure. Structures D-G may form nonreversible
covalent bonds with reactive nucleophilic groups in a combining
site of an antibody. In these structures, R''.sub.1 and R''.sub.2
represent C, 0, N, halide or leaving groups such as mesyl or
tosyl.
[0244] FIG. 9 shows various electrophiles that are suitable for
reactive modification with a reactive amino acid side chain in a
combining site of an antibody and thus may serve as linker reactive
groups. Key: (A) acyl beta-lactam; (B) simple diketone; (C)
succinimide active ester; (D) maleimide; (E) haloacetamide with
linker; (F) haloketone; (G) cyclohexyl diketone; and (H) aldehyde.
The squiggle line indicates the point of attachment to the rest of
the linker or targeting agent. X refers to a halogen.
[0245] FIG. 10 shows the addition of a nucleophilic ("nu") side
chain in an antibody combining site to compounds A-G in FIG. 8.
Antibody-Nu- refers to a covalent bond to an amino acid side chain
bearing a nucleophile in a combining site of an antibody.
[0246] FIG. 11 shows the addition of a nucleophilic side chain in
an antibody combining to compounds A-H in FIG. 9. Antibody-Nu-
refers to a covalent bond to an amino acid side chain bearing a
nucleophile in a combining site of an antibody.
[0247] FIG. 12 shows a synthesis of:
##STR00011##
[0248] FIG. 13 shows a synthesis of:
##STR00012##
[0249] FIG. 14 shows a synthesis of:
##STR00013##
[0250] FIG. 15 shows a synthesis of:
##STR00014##
[0251] FIG. 16 shows a synthesis of:
##STR00015##
[0252] FIG. 17 shows a synthesis of:
##STR00016##
[0253] FIG. 18 shows a synthesis of:
##STR00017##
[0254] FIG. 19 shows a synthesis of:
##STR00018##
[0255] FIG. 20 shows a synthesis of:
##STR00019##
[0256] FIG. 21 shows syntheses of:
##STR00020##
[0257] FIG. 22 shows a synthesis of:
##STR00021##
[0258] FIG. 23 shows a synthesis of:
##STR00022##
[0259] FIG. 24 shows a synthesis of:
##STR00023##
[0260] FIG. 25 shows a synthesis of:
##STR00024##
[0261] FIG. 26 shows a synthesis of:
##STR00025##
[0262] FIG. 27 shows a synthesis of:
##STR00026##
[0263] FIG. 28 shows a synthesis of:
##STR00027##
[0264] FIG. 29 illustrates one embodiment of the invention.
[0265] FIG. 30 illustrates one embodiment of the invention.
[0266] FIG. 31 illustrates one embodiment of the invention.
[0267] FIG. 32 illustrates one embodiment of the invention.
[0268] FIG. 33 illustrates one embodiment of the invention.
[0269] FIG. 34 illustrates one embodiment of the invention.
[0270] FIG. 35 illustrates one embodiment of the invention.
[0271] FIG. 36 illustrates one embodiment of the invention.
[0272] FIG. 37 illustrates one embodiment of the invention.
[0273] FIG. 38 illustrates one embodiment of the invention
[0274] FIG. 39 illustrates one embodiment of the invention.
[0275] FIG. 40 illustrates one embodiment of the invention.
[0276] FIG. 41 illustrates one embodiment of the invention.
DETAILED DESCRIPTION
Definitions
[0277] The following abbreviations, terms and phrases are used
herein as defined below.
TABLE-US-00020 TABLE 1 Amino acid abbreviations One letter Three
letter Amino acid abbreviation or other abbreviation Alanine A Ala
Arginine R Arg Asparagine N Asn Aspartic acid D Asp Cysteine C Cys
Glutamic acid E Glu Glutamine Q Gln Glycine G Gly Histidine H His
Isoleucine I Ile Leucine L Leu Lysine K Lys Methionine M Met
Phenylalanine F Phe Proline P Pro Serine S Ser Threonine T Thr
Tryptophan W Trp Tyrosine Y Tyr Valine V Val Dehydroproline -- DHP
Hydroxyproline -- HP Homocyclohexyl alanine -- HChA Homophenyl
alanine HF Thiazolyl alanine ThA Norvaline Nva Homoleucine HL
Epsilon acyl lysine AcK 4-benzoyl phenylalanine BPA 4-Carboxy
phenylalanine (CO2H)F: also: CF Epsilon chloro benzyl
(CIBnCarbamate)K: also: CbcK carbamate lysine Diaminobutyric acid
Dab Diaminopropionic acid Dap 4-Nitro phenylalanine NO2F: also: NF
(2S,4R)-4-Hydroxyproline BnHP Nictinyl lysine NicK Thienyl Alanine
TA
[0278] Every amino-bearing side chain of a targeting agent can be
terminated by R.sup.1 as defined herein. Every
COOH/COO.sup.--bearing side chain of a targeting agent can be
terminated by R.sup.2 as defined herein.
##STR00028## ##STR00029## ##STR00030## ##STR00031##
shown attached to a lysine residue.
[0279] Unless indicated otherwise by a "D" prefix, e.g., D-Ala or
N-Me-D-Ile, the stereochemistry of the alpha-carbon of the amino
acids and aminoacyl residues in peptides described in this
specification and the appended claims is the natural or "L"
configuration. The Cahn-Ingold-Prelog "R" and "S" designations are
used to specify the stereochemistry of chiral centers in certain
acyl substituents at the N-terminus of the peptides. The
designation "R,S" is meant to indicate a racemic mixture of the two
enantiomeric forms. This nomenclature follows that described in R.
S. Cahn, et al., Angew. Chem. Int. Ed. Engl., 5:385-415 (1966).
[0280] "Polypeptide," "peptide," and "protein" are used
interchangeably to refer to a polymer of amino acid residues. As
used herein, these terms apply to amino acid polymers in which one
or more amino acid residues is an artificial chemical analog of a
corresponding naturally occurring amino acid. These terms also
apply to naturally occurring amino acid polymers. Amino acids can
be in the L or D form as long as the binding function of the
peptide is maintained. Peptides may be cyclic, having an
intramolecular bond between two non-adjacent amino acids within the
peptide, e.g., backbone to backbone, side-chain to backbone and
side-chain to side-chain cyclization. Cyclic peptides can be
prepared by methods well know in the art. See e.g., U.S. Pat. No.
6,013,625.
[0281] All peptide sequences are written according to the generally
accepted convention whereby the alpha-N-terminal amino acid residue
is on the left and the alpha-C-terminal amino acid residue is on
the right. As used herein, the term "N-terminus" refers to the free
alpha-amino group of an amino acid in a peptide, and the term
"C-terminus" refers to the free .alpha.-carboxylic acid terminus of
an amino acid in a peptide. A peptide which is N-terminated with a
group refers to a peptide bearing a group on the alpha-amino
nitrogen of the N-terminal amino acid residue. An amino acid which
is N-terminated with a group refers to an amino acid bearing a
group on the alpha-amino nitrogen.
[0282] "Substantially homologous" means at least about 75%
(preferably at least about 80%, and more preferably at least about
90% or most preferably at least about 95%, of the amino-acid
residues match over the defined length of the peptide sequences.
Substantially homologous sequences include sequences sharing at
least 65% of the amino acid residues over their length, and at
least 10% of the non-shared sequences being conservative
subustitutions. Sequences that are substantially homologous can be
identified by comparing the sequences using standard software
available in sequence data banks, such as BLAST programs available
from the National Cancer Center for Biotechology Information at
ncbi.nlm.nih.gov.
[0283] In general, "substituted" refers to a group as defined below
in which one or more bonds to a hydrogen atom contained therein are
replaced by a bond to non-hydrogen or non-carbon atoms such as, but
not limited to, a halogen atom such as F, Cl, Br, and I; an oxygen
atom in groups such as hydroxyl groups, alkoxy groups, aryloxy
groups, and ester groups; a sulfur atom in groups such as thiol
groups, alkyl and aryl sulfide groups, sulfone groups, sulfonyl
groups, and sulfoxide groups; a nitrogen atom in groups such as
amines, amides, alkylamines, dialkylamines, arylamines,
alkylarylamines, diarylamines, N-oxides, imides, and enamines; a
silicon atom in groups such as in trialkylsilyl groups,
dialkylarylsilyl groups, alkyldiarylsilyl groups, and triarylsilyl
groups; and other heteroatoms in various other groups. Substituted
alkyl groups and also substituted cycloalkyl groups and others also
include groups in which one or more bonds to a carbon(s) or
hydrogen(s) atom is replaced by a bond to a heteroatom such as
oxygen in carbonyl, carboxyl, and ester groups; nitrogen in groups
such as imines, oximes, hydrazones, and nitriles. As employed
herein, a group which is "optionally substituted" may be
substituted or unsubstituted. Thus, e.g., "optionally substituted
alkyl" refers to both substituted alkyl groups and unsubstituted
alkyl groups.
[0284] The phrase "unsubstituted alkyl" refers to alkyl groups that
do not contain heteroatoms. Thus, the phrase includes straight
chain alkyl groups such as methyl, ethyl, propyl, butyl, pentyl,
hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl and the like.
The phrase also includes branched chain isomers of straight chain
alkyl groups, including but not limited to, the following which are
provided by way of example: --CH(CH.sub.3).sub.2,
--CH(CH.sub.3)(CH.sub.2CH.sub.3), --CH(CH.sub.2CH.sub.3).sub.2,
--C(CH.sub.3).sub.3, --C(CH.sub.2CH.sub.3).sub.3,
--CH.sub.2CH(CH.sub.3).sub.2,
--CH.sub.2CH(CH.sub.3)(CH.sub.2CH.sub.3),
--CH.sub.2CH(CH.sub.2CH.sub.3).sub.2, --CH.sub.2C(CH.sub.3).sub.3,
--CH.sub.2C(CH.sub.2CH.sub.3).sub.3,
--CH(CH.sub.3)CH(CH.sub.3)(CH.sub.2CH.sub.3),
--CH.sub.2CH.sub.2CH(CH.sub.3).sub.2,
--CH.sub.2CH.sub.2CH(CH.sub.3)(CH.sub.2CH.sub.3),
--CH.sub.2CH.sub.2CH(CH.sub.2CH.sub.3).sub.2,
--CH.sub.2CH.sub.2C(CH.sub.3).sub.3,
--CH.sub.2CH.sub.2C(CH.sub.2CH.sub.3).sub.3,
--CH(CH.sub.3)CH.sub.2CH(CH.sub.3).sub.2,
--CH(CH.sub.3)CH(CH.sub.3)CH(CH.sub.3).sub.2,
--CH(CH.sub.2CH.sub.3)CH(CH.sub.3)CH(CH.sub.3)(CH.sub.2CH.sub.3),
and others. The phrase does not include cycloalkyl groups. Thus,
the phrase unsubstituted alkyl groups includes primary alkyl
groups, secondary alkyl groups, and tertiary alkyl groups.
Unsubstituted alkyl groups may be bonded to one or more carbon
atom(s), oxygen atom(s), nitrogen atom(s), and/or sulfur atom(s) in
the parent compound. Possible unsubstituted alkyl groups include
straight and branched chain alkyl groups having 1 to 20 carbon
atoms. Alternatively, such unsubstituted alkyl groups have from 1
to 10 carbon atoms or are lower alkyl groups having from 1 to about
6 carbon atoms. Other unsubstituted alkyl groups include straight
and branched chain alkyl groups having from 1 to 3 carbon atoms and
include methyl, ethyl, propyl, and --CH(CH.sub.3).sub.2.
[0285] The phrase "substituted alkyl" refers to an alkyl group in
which one or more bonds to a carbon(s) or hydrogen(s) are replaced
by a bond to non-hydrogen and non-carbon atoms such as, but not
limited to, a halogen atom in halides such as F, Cl, Br, and I; an
oxygen atom in groups such as hydroxyl groups, alkoxy groups,
aryloxy groups, and ester groups; a sulfur atom in groups such as
thiol groups, alkyl and aryl sulfide groups, sulfone groups,
sulfonyl groups, and sulfoxide groups; a nitrogen atom in groups
such as amines, amides, alkylamines, dialkylamines, arylamines,
alkylarylamines, diarylamines, N-oxides, imides, and enamines; a
silicon atom in groups such as in trialkylsilyl groups,
dialkylarylsilyl groups, alkyldiarylsilyl groups, and triarylsilyl
groups; and other heteroatoms in various other groups. Substituted
alkyl groups also include groups in which one or more bonds to a
carbon(s) or hydrogen(s) atom is replaced by a bond to a heteroatom
such as oxygen in carbonyl, carboxyl, and ester groups; nitrogen in
groups such as imines, oximes, hydrazones, and nitriles.
Substituted alkyl groups include, among others, alkyl groups in
which one or more bonds to a carbon or hydrogen atom is/are
replaced by one or more bonds to fluorine atoms. One example of a
substituted alkyl group is the trifluoromethyl group and other
alkyl groups that contain the trifluoromethyl group. Other alkyl
groups include those in which one or more bonds to a carbon or
hydrogen atom is replaced by a bond to an oxygen atom such that the
substituted alkyl group contains a hydroxyl, alkoxy, aryloxy group,
or heterocyclyloxy group. Still other alkyl groups include alkyl
groups that have an amine, alkylamine, dialkylamine, arylamine,
(alkyl)(aryl)amine, diarylamine, heterocyclylamine,
(alkyl)(heterocyclyl)amine, (aryl)(heterocyclyl)amine, or
diheterocyclylamine group.
[0286] The phrase "unsubstituted alkylene" refers to a divalent
unsubstituted alkyl group as defined above. Thus methylene,
ethylene, and propylene are each examples of unsubstituted
alkylenes. The phrase "substituted alkylene" refers to a divalent
substituted alkyl group as defined above. Substituted or
unsubstituted lower alkylene groups have from 1 to about 6
carbons.
[0287] The phrase "unsubstituted cycloalkyl" refers to cyclic alkyl
groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, and cyclooctyl and such rings substituted with
straight and branched chain alkyl groups as defined above. The
phrase also includes polycyclic alkyl groups such as, but not
limited to, adamantyl norbornyl, and bicyclo[2.2.2]octyl and the
like, as well as such rings substituted with straight and branched
chain alkyl groups as defined above. Thus, the phrase would include
methylcylcohexyl groups among others. The phrase does not include
cyclic alkyl groups containing heteroatoms. Unsubstituted
cycloalkyl groups may be bonded to one or more carbon atom(s),
oxygen atom(s), nitrogen atom(s), and/or sulfur atom(s) in the
parent compound. In some embodiments unsubstituted cycloalkyl
groups have from 3 to 20 carbon atoms. In other embodiments, such
unsubstituted alkyl groups have from 3 to 8 carbon atoms while in
others, such groups have from 3 to 7 carbon atoms.
[0288] The phrase "substituted cycloalkyl" "has the same meaning
with respect to unsubstituted cycloalkyl groups that substituted
alkyl groups have with respect to unsubstituted alkyl groups. Thus,
the phrase includes, but is not limited to, oxocyclohexyl,
chlorocyclohexyl, hydroxycyclopentyl, and chloromethylcyclohexyl
groups.
[0289] The phrase "unsubstituted aryl" refers to aryl groups that
do not contain heteroatoms. Thus the phrase includes, but is not
limited to, groups such as phenyl, biphenyl, anthracenyl, and
naphthenyl by way of example. Although the phrase "unsubstituted
aryl" includes groups containing condensed rings such as
naphthalene, it does not include aryl groups that have other groups
such as alkyl or halo groups bonded to one of the ring members, as
aryl groups such as tolyl are considered herein to be substituted
aryl groups as described below. Typically, an unsubstituted aryl
may be a lower aryl, having from 6 to about 10 carbon atoms. One
unsubstituted aryl group is phenyl. Unsubstituted aryl groups may
be bonded to one or more carbon atom(s), oxygen atom(s), nitrogen
atom(s), and/or sulfur atom(s) in the parent compound, however.
[0290] The phrase "substituted aryl group" has the same meaning
with respect to unsubstituted aryl groups that substituted alkyl
groups have with respect to unsubstituted alkyl groups. However, a
substituted aryl group also includes aryl groups in which one of
the aromatic carbons is bonded to one of the non-carbon or
non-hydrogen atoms described above and also includes aryl groups in
which one or more aromatic carbons of the aryl group is bonded to a
substituted and/or unsubstituted alkyl, alkenyl, or alkynyl group
as defined herein. This includes bonding arrangements in which two
carbon atoms of an aryl group are bonded to two atoms of an alkyl,
alkenyl, or alkynyl group to define a fused ring system (e.g.,
dihydronaphthyl or tetrahydronaphthyl). Thus, the phrase
"substituted aryl" includes, but is not limited to tolyl and
hydroxyphenyl among others.
[0291] The phrase "unsubstituted alkenyl" refers to straight and
branched chain and cyclic groups such as those described with
respect to unsubstituted alkyl groups as defined above, except that
at least one double bond exists between two carbon atoms. Examples
include, but are not limited to vinyl, --CH.dbd.C(H)(CH.sub.3),
--CH.dbd.C(CH.sub.3).sub.2, --C(CH.sub.3).dbd.C(H).sub.2,
--C(CH.sub.3).dbd.C(H)(CH.sub.3),
--C(CH.sub.2CH.sub.3).dbd.CH.sub.2, cyclohexenyl, cyclopentenyl,
cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl among
others. Lower unsubstituted alkenyl groups have from 1 to about 6
carbons.
[0292] The phrase "substituted alkenyl" has the same meaning with
respect to unsubstituted alkenyl groups that substituted alkyl
groups have with respect to unsubstituted alkyl groups. A
substituted alkenyl group includes alkenyl groups in which a
non-carbon or non-hydrogen atom is bonded to a carbon double bonded
to another carbon and those in which one of the non-carbon or
non-hydrogen atoms is bonded to a carbon not involved in a double
bond to another carbon. For example, --CH.dbd.CH--OCH.sub.3 and
--CH.dbd.CH--CH.sub.2--CH are both substituted alkenyls.
Oxoalkenyls wherein a CH.sub.2 group is replaced by a carbonyl,
such as --CH.dbd.CH--C(O)--CH.sub.3, are also substituted
alkenyls.
[0293] The phrase "unsubstituted alkenylene" refers to a divalent
unsubstituted alkenyl group as defined above. For example,
--CH.dbd.CH-- is an exemplary unsubstituted alkenylene. The phrase
"substituted alkenylene" refers to a divalent substituted alkenyl
group as defined above.
[0294] The phrase "unsubstituted alkynyl" refers to straight and
branched chain groups such as those described with respect to
unsubstituted alkyl groups as defined above, except that at least
one triple bond exists between two carbon atoms. Examples include,
but are not limited to, --C.ident.C(H), --C.ident.C(CH.sub.3),
--C.ident.C(CH.sub.2CH.sub.3), --C(H.sub.2)C.ident.C(H),
--C(H).sub.2C.ident.C(CH.sub.3), and
--C(H).sub.2C.ident.C(CH.sub.2CH.sub.3) among others. Unsubstituted
lower alkynyl groups have from 1 to about 6 carbons.
[0295] The phrase "substituted alkynyl" has the same meaning with
respect to unsubstituted alkynyl groups that substituted alkyl
groups have with respect to unsubstituted alkyl groups. A
substituted alkynyl group includes alkynyl groups in which a
non-carbon or non-hydrogen atom is bonded to a carbon triple bonded
to another carbon and those in which a non-carbon or non-hydrogen
atom is bonded to a carbon not involved in a triple bond to another
carbon. Examples include, but are not limited to, oxoalkynyls
wherein a CH.sub.2 group is replaced by a carbonyl, such as
--C(O)--CH.ident.CH--CH.sub.3 and
--C(O)--CH.sub.2--CH.ident.CH.
[0296] The phrase "unsubstituted alkynylene" refers to a divalent
unsubstituted alkynyl group as defined above. A --C.ident.C-- is an
example of an unsubstituted alkynylene. The phrase "substituted
alkynylene" refers to a divalent substituted alkynyl group as
defined above.
[0297] The phrase "unsubstituted aralkyl" refers to unsubstituted
alkyl groups as defined above in which a hydrogen or carbon bond of
the unsubstituted alkyl group is replaced with a bond to an aryl
group as defined above. For example, methyl (--CH.sub.3) is an
unsubstituted alkyl group. If a hydrogen atom of the methyl group
is replaced by a bond to a phenyl group, such as if the carbon of
the methyl were bonded to a carbon of benzene, then the compound is
an unsubstituted aralkyl group (i.e., a benzyl group). Thus, the
phrase includes, but is not limited to, groups such as benzyl,
diphenylmethyl, and 1-phenylethyl
(--CH(C.sub.6H.sub.5)(CH.sub.3)).
[0298] The phrase "substituted aralkyl" has the same meaning with
respect to unsubstituted aralkyl groups that substituted aryl
groups have with respect to unsubstituted aryl groups. However,
substituted aralkyls also include groups in which a carbon or
hydrogen bond of the alkyl part of the group is replaced by a bond
to a non-carbon or a non-hydrogen atom. Examples of substituted
aralkyl groups include, but are not limited to,
--CH.sub.2C(.dbd.O)(C.sub.6H.sub.5), and
--CH.sub.2(2-methylphenyl).
[0299] The phrase "unsubstituted aralkenyl" refers to unsubstituted
alkenyl groups as defined above in which a hydrogen or carbon bond
of the unsubstituted alkenyl group is replaced with a bond to an
aryl group as defined above. For example, vinyl is an unsubstituted
alkenyl group. If a hydrogen atom of the vinyl group is replaced by
a bond to a phenyl group, such as if a carbon of the vinyl were
bonded to a carbon of benzene, then the compound is an
unsubstituted aralkenyl group (i.e., a styryl group). Thus, the
phrase includes, but is not limited to, groups such as styryl,
diphenylvinyl, and 1-phenylethenyl
(--C(C.sub.6H.sub.5)(CH.sub.2)).
[0300] The phrase "substituted aralkenyl" has the same meaning with
respect to unsubstituted aralkenyl groups that substituted aryl
groups have with respect to unsubstituted aryl groups. A
substituted aralkenyl group also includes groups in which a carbon
or hydrogen bond of the alkenyl part of the group is replaced by a
bond to a non-carbon or a non-hydrogen atom. Examples of
substituted aralkenyl groups include, but are not limited to,
--CH.dbd.C(Cl)(C.sub.6H.sub.5), and
--CH.dbd.CH(2-methylphenyl).
[0301] The phrase "unsubstituted aralkynyl" refers to unsubstituted
alkynyl groups as defined above in which a hydrogen or carbon bond
of the unsubstituted alkynyl group is replaced with a bond to an
aryl group as defined above. For example, acetylene is an
unsubstituted alkynyl group. If a hydrogen atom of the acetylene
group is replaced by a bond to a phenyl group, such as if a carbon
of the acetylene were bonded to a carbon of benzene, then the
compound is an unsubstituted aralkynyl group. Thus, the phrase
includes, but is not limited to, groups such as --C.ident.C-phenyl
and --CH.sub.2--C.ident.C-phenyl.
[0302] The phrase "substituted aralkynyl" has the same meaning with
respect to unsubstituted aralkynyl groups that substituted aryl
groups have with respect to unsubstituted aryl groups. However, a
substituted aralkynyl group also includes groups in which a carbon
or hydrogen bond of the alkynyl part of the group is replaced by a
bond to a non-carbon or a non-hydrogen atom. Examples of
substituted aralkynyl groups include, but are not limited to,
--C.ident.C--C(Br)(C.sub.6H.sub.5) and
--C.ident.C(2-methylphenyl).
[0303] The phrase "unsubstituted heteroalkyl" refers to
unsubstituted alkyl groups as defined above in which the carbon
chain is interrupted by one or more heteroatoms chosen from N, O,
and S. Unsubstituted heteroalkyls containing N may have NH or
N(unsubstituted alkyl) in the carbon chain. For example,
unsubstituted heteroalkyls include alkoxy, alkoxyalkyl,
alkoxyalkoxy, thioether, alkylaminoalkyl, aminoalkyloxy, and other
such groups. Typically, unsubstituted heteroalkyl groups contain
1-5 heteroatoms, and particularly 1-3 heteroatoms. In some
embodiments unsubstituted heteroalkyls include, for example,
alkoxyalkoxyalkoxy groups such as ethyloxyethyloxyethyloxy.
[0304] The phrase "substituted heteroalkyl" has the same meaning
with respect to unsubstituted heteroalkyl groups that substituted
alkyl groups have with respect to unsubstituted alkyl groups.
[0305] The phrase "unsubstituted heteroalkylene" refers to a
divalent unsubstituted heteroalkyl group as defined above. For
example, --CH.sub.2--O--CH.sub.2-- and
--CH.sub.2--NH--CH.sub.2CH.sub.2-- are both exemplary unsubstituted
heteroalkylenes. The phrase "substituted heteroalkylene" refers to
a divalent substituted heteroalkyl group As defined above.
[0306] The phrase "unsubstituted heteroalkenyl" refers to
unsubstituted alkene groups as defined above in which the carbon
chain is interrupted by one or more heteroatoms chosen from N, O,
and S. Unsubstituted heteroalkenyls containing N may have NH or
N(unsubstituted alkyl or alkene) in the carbon chain. The phrase
"substituted heteroalkenyl" has the same meaning with respect to
unsubstituted heteroalkenyl groups that substituted heteroalkyl
groups have with respect to unsubstituted heteroalkyl groups.
[0307] The phrase "unsubstituted heteroalkenylene" refers to a
divalent unsubstituted heteroalkenyl group as defined above. Thus
--CH.sub.2--O--CH.dbd.CH-- is an example of an unsubstituted
heteroalkenylene. The phrase "substituted heteroalkenylene" refers
to a divalent substituted heteroalkenyl group as defined above.
[0308] The phrase "unsubstituted heteroalkynyl" refers to
unsubstituted alkynyl groups as defined above in which the carbon
chain is interrupted by one or more heteroatoms chosen from N, O,
and S. Unsubstituted heteroalkynyls containing N may have NH or
N(unsubstituted alkyl, alkene, or alkyne) in the carbon chain. The
phrase "substituted heteroalkynyl" has the same meaning with
respect to unsubstituted heteroalkynyl groups that substituted
heteroalkyl groups have with respect to unsubstituted heteroalkyl
groups.
[0309] The phrase "unsubstituted heteroalkynylene" refers to a
divalent unsubstituted heteroalkynyl group as defined above. Thus
--CH.sub.2--O--CH.sub.2--C.ident.C-- is an example of an
unsubstituted heteroalkynylene. The phrase "substituted
heteroalkynylene" refers to a divalent substituted heteroalkynyl
group as defined above.
[0310] The phrase "unsubstituted heterocyclyl" refers to both
aromatic and nonaromatic ring compounds including monocyclic,
bicyclic, and polycyclic ring compounds such as, but not limited
to, quinuclidyl, containing 3 or more ring members of which one or
more is a heteroatom such as, but not limited to, N, O, and S.
Although the phrase "unsubstituted heterocyclyl" includes condensed
heterocyclic rings such as benzimidazolyl, it does not include
heterocyclyl groups that have other groups such as alkyl or halo
groups bonded to one of the ring members as compounds such as
2-methylbenzimidazolyl are substituted heterocyclyl groups.
Examples of heterocyclyl groups include, but are not limited to:
unsaturated 3 to 8 membered rings containing 1 to 4 nitrogen atoms
such as, but not limited to pyrrolyl, pyrrolinyl, imidazolyl,
pyrazolyl, pyridyl, dihydropyridyl, pyrimidyl, pyrazinyl,
pyridazinyl, triazolyl (e.g., 4H-1,2,4-triazolyl,
1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl etc.), tetrazolyl, (e.g.,
1H-tetrazolyl, 2H tetrazolyl, etc.); saturated 3 to 8 membered
rings containing 1 to 4 nitrogen atoms such as, but not limited to,
pyrrolidinyl, imidazolidinyl, piperidinyl, piperazinyl; condensed
unsaturated heterocyclic groups containing 1 to 4 nitrogen atoms
such as, but not limited to, indolyl, isoindolyl, indolinyl,
indolizinyl, benzimidazolyl, quinolyl, isoquinolyl, indazolyl,
benzotriazolyl; unsaturated 3 to 8 membered rings containing 1 to 2
oxygen atoms and 1 to 3 nitrogen atoms such as, but not limited to,
oxazolyl, isoxazolyl, oxadiazolyl (e.g., 1,2,4-oxadiazolyl,
1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl, etc.); saturated 3 to 8
membered rings containing 1 to 2 oxygen atoms and 1 to 3 nitrogen
atoms such as, but not limited to, morpholinyl; unsaturated
condensed heterocyclic groups containing 1 to 2 oxygen atoms and 1
to 3 nitrogen atoms, for example, benzoxazolyl, benzoxadiazolyl,
benzoxazinyl (e.g., 2H-1,4-benzoxazinyl, etc.); unsaturated 3 to 8
membered rings containing 1 to 3 sulfur atoms and 1 to 3 nitrogen
atoms such as, but not limited to, thiazolyl, isothiazolyl,
thiadiazolyl (e.g., 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl,
1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, etc.); saturated 3 to 8
membered rings containing 1 to 2 sulfur atoms and 1 to 3 nitrogen
atoms such as, but not limited to, thiazolodinyl; saturated and
unsaturated 3 to 8 membered rings containing 1 to 2 sulfur atoms
such as, but not limited to, thienyl, dihydrodithiinyl,
dihydrodithionyl, tetrahydrothiophene, tetrahydrothiopyran;
unsaturated condensed heterocyclic rings containing 1 to 2 sulfur
atoms and 1 to 3 nitrogen atoms such as, but not limited to,
benzothiazolyl, benzothiadiazolyl, benzothiazinyl (e.g.,
2H-1,4-benzothiazinyl, etc.), dihydrobenzothiazinyl (e.g.,
2H-3,4-dihydrobenzothiazinyl, etc.), unsaturated 3 to 8 membered
rings containing oxygen atoms such as, but not limited to furyl;
unsaturated condensed heterocyclic rings containing 1 to 2 oxygen
atoms such as benzodioxolyl (e.g., 1,3-benzodioxoyl, etc.);
unsaturated 3 to 8 membered rings containing an oxygen atom and 1
to 2 sulfur atoms such as, but not limited to, dihydrooxathiinyl;
saturated 3 to 8 membered rings containing 1 to 3 oxygen atoms and
1 to 2 sulfur atoms such as 1,4-oxathiane; unsaturated condensed
rings containing 1 to 2 sulfur atoms such as benzothienyl,
benzodithiinyl; and unsaturated condensed heterocyclic rings
containing an oxygen atom and 1 to 3 oxygen atoms such as
benzoxathiinyl. Heterocyclyl group also include those described
above in which one or more S atoms in the ring is double-bonded to
one or two oxygen atoms (sulfoxides and sulfones). For example,
heterocyclyl groups include tetrahydrothiophene,
tetrahydrothiophene oxide, and tetrahydrothiophene 1,1-dioxide. In
some embodiments heterocyclyl groups contain 5 or 6 ring members.
In other embodiments heterocyclyl groups include morpholine,
piperazine, piperidine, pyrrolidine, imidazole, pyrazole,
1,2,3-triazole, 1,2,4-triazole, tetrazole, thiomorpholine,
thiomorpholine in which the S atom of the thiomorpholine is bonded
to one or more 0 atoms, pyrrole, homopiperazine, oxazolidin-2-one,
pyrrolidin-2-one, oxazole, quinuclidine, thiazole, isoxazole,
furan, and tetrahydrofuran.
[0311] The phrase "substituted heterocyclyl" refers to an
unsubstituted heterocyclyl group as defined above in which one of
the ring members is bonded to a non-hydrogen atom such as described
above with respect to substituted alkyl groups and substituted aryl
groups. Examples include, but are not limited to,
2-methylbenzimidazolyl, 5-methylbenzimidazolyl,
5-chlorobenzthiazolyl, 1-methyl piperazinyl, and
2-chloropyridyl.
[0312] The phrase "unsubstituted heteroaryl" refers to
unsubstituted aromatic heterocyclyl groups as defined above. Thus,
unsubstituted heteroaryl groups include but are not limited to
furyl, imidazolyl, oxazolyl, isoxazolyl, pyridinyl, benzimidazolyl,
and benzothiazolyl. The phrase "substituted heteroaryl" refers to
substituted aromatic heterocyclyl groups as defined above.
[0313] The phrase "unsubstituted heterocyclylalkyl" refers to
unsubstituted alkyl groups as defined above in which a hydrogen or
carbon bond of the unsubstituted alkyl group is replaced with a
bond to a heterocyclyl group as defined above. For example, methyl
(--CH.sub.3) is an unsubstituted alkyl group. If a hydrogen atom of
the methyl group is replaced by a bond to a heterocyclyl group,
such as if the carbon of the methyl were bonded to carbon 2 of
pyridine (one of the carbons bonded to the N of the pyridine) or
carbons 3 or 4 of the pyridine, then the compound is an
unsubstituted heterocyclylalkyl group.
[0314] The phrase "substituted heterocyclylalkyl" has the same
meaning with respect to unsubstituted heterocyclylalkyl groups that
substituted aralkyl groups have with respect to unsubstituted
aralkyl groups. A substituted heterocyclylalkyl group also includes
groups in which a non-hydrogen atom is bonded to a heteroatom in
the heterocyclyl group of the heterocyclylalkyl group such as, but
not limited to, a nitrogen atom in the piperidine ring of a
piperidinylalkyl group.
[0315] The phrase "unsubstituted heterocyclylalkenyl" refers to
unsubstituted alkenyl groups as defined above in which a hydrogen
or carbon bond of the unsubstituted alkenyl group is replaced with
a bond to a heterocyclyl group as defined above. For example, vinyl
is an unsubstituted alkenyl group. If a hydrogen atom of the vinyl
group is replaced by a bond to a heterocyclyl group, such as if the
carbon of the vinyl were bonded to carbon 2 of pyridine or carbons
3 or 4 of the pyridine, then the compound is an unsubstituted
heterocyclylalkenyl group.
[0316] The phrase "substituted heterocyclylalkenyl" has the same
meaning with respect to unsubstituted heterocyclylalkenyl groups
that substituted aralkenyl groups have with respect to
unsubstituted aralkenyl groups. However, a substituted
heterocyclylalkenyl group also includes groups in which a
non-hydrogen atom is bonded to a heteroatom in the heterocyclyl
group of the heterocyclylalkenyl group such as, but not limited to,
a nitrogen atom in the piperidine ring of a piperidinylalkenyl
group.
[0317] The phrase "unsubstituted heterocyclylalkynyl" refers to
unsubstituted alkynyl groups as defined above in which a hydrogen
or carbon bond of the unsubstituted alkynyl group is replaced with
a bond to a heterocyclyl group as defined above. For example,
acetylene is an unsubstituted alkynyl group. If a hydrogen atom of
the acetylene group is replaced by a bond to a heterocyclyl group,
such as if the carbon of the acetylene were bonded to carbon 2 of
pyridine or carbons 3 or 4 of the pyridine, then the compound is an
unsubstituted heterocyclylalkynyl group.
[0318] The phrase "substituted heterocyclylalkynyl" has the same
meaning with respect to unsubstituted heterocyclylalkynyl groups
that substituted aralkynyl groups have with respect to
unsubstituted aralkynyl groups. A substituted heterocyclylalkynyl
group also includes groups in which a non-hydrogen atom is bonded
to a heteroatom in the heterocyclyl group of the
heterocyclylalkynyl group such as, but not limited to, a nitrogen
atom in the piperidine ring of a piperidinylalkynyl group.
[0319] The phrase "unsubstituted alkoxy" refers to a hydroxyl group
(--OH) in which the bond to the hydrogen atom is replaced by a bond
to a carbon atom of an otherwise unsubstituted alkyl group as
defined above.
[0320] The phrase "substituted alkoxy" refers to a hydroxyl group
(--OH) in which the bond to the hydrogen atom is replaced by a bond
to a carbon atom of an otherwise substituted alkyl group as defined
above.
[0321] A "pharmaceutically acceptable salt" includes a salt with an
inorganic base, organic base, inorganic acid, organic acid, or
basic or acidic amino acid. Salts of inorganic bases include, for
example, alkali metals such as sodium or potassium; alkaline earth
metals such as calcium and magnesium or aluminum; and ammonia.
Salts of organic bases include, for example, trimethylamine,
triethylamine, pyridine, picoline, ethanolamine, diethanolamine,
and triethanolamine. Salts of inorganic acids include for example,
hydrochloric acid, hydroboric acid, nitric acid, sulfuric acid, and
phosphoric acid. Salts of organic acids include for example, formic
acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid,
tartaric acid, maleic acid, citric acid, succinic acid, malic acid,
methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic
acid. Salts of basic amino acids include, for example, arginine,
lysine and ornithine. Acidic amino acids include, for example,
aspartic acid and glutamic acid.
[0322] "Tautomers" refers to isomeric forms of a compound that are
in equilibrium with each other. The concentrations of the isomeric
forms will depend on the environment the compound is found in and
may be different depending upon, for example, whether the compound
is a solid or is in an organic or aqueous solution. For example, in
aqueous solution, ketones are typically in equilibrium with their
enol forms. Thus, ketones and their enols are referred to as
tautomers of each other. As readily understood by one skilled in
the art, a wide variety of functional groups and other structures
may exhibit tautomerism, and all tautomers of compounds of Formulas
I, II, and III are within the scope of the present invention.
[0323] The compounds according to the invention may be solvated,
especially hydrated. Hydration may occur during manufacturing of
the compounds or compositions comprising the compounds, or the
hydration may occur over time due to the hygroscopic nature of the
compounds.
[0324] Certain embodiments are derivatives referred to as prodrugs.
The expression "prodrug" denotes a derivative of a pharmaceutically
or therapeutically active drug, e.g., esters and amides, wherein
the derivative has an enhanced characteristic such as, for example,
enhanced delivery and therapeutic value as compared to the drug and
can be transformed into the drug by an enzymatic or chemical
process. See, for example, R. E. Notari, Methods Enzymol.
112:309-323 (1985); N. Bodor, Drugs of the Future 6:165-182 (1981);
H. Bundgaard, Chapter 1 in Design of Prodrugs (H. Bundgaard, ed.),
Elsevier, New York (1985); and A. G. Gilman et al., Goodman And
Gilman's The Pharmacological Basis of Therapeutics, 8.sup.th ed.,
McGraw-Hill (1990). Thus, the prodrug may be designed to alter the
metabolic stability or transport characteristics of a drug, mask
side effects or toxicity of a drug, improve the flavor of a drug,
or to alter other characteristics or properties of a drug.
[0325] Compounds of the present invention include enriched or
resolved optical isomers at any or all asymmetric atoms as are
apparent from the depictions. Both racemic and diastereomeric
mixtures, as well as the individual optical isomers can be isolated
or synthesized so as to be substantially free of their enantiomeric
or diastereomeric partners. All such stereoisomers are within the
scope of the invention.
[0326] The term "carboxy protecting group" as used herein refers to
a carboxylic acid protecting ester group employed to block or
protect the carboxylic acid functionality while the reactions
involving other functional sites of the compound are carried out.
Carboxy protecting groups are disclosed in, for example, Greene,
Protective Groups in Organic Synthesis, pp. 152-186, John Wiley
& Sons, New York (1981), which is hereby incorporated herein by
reference. In addition, a carboxy protecting group can be used as a
prodrug, whereby the carboxy protecting group can be readily
cleaved in vivo by, for example, enzymatic hydrolysis to release
the biologically active parent. T. Higuchi and V. Stella provide a
discussion of the prodrug concept in "Pro-drugs as Novel Delivery
Systems", Vol. 14 of the A.C.S. Symposium Series, American Chemical
Society (1975), which is hereby incorporated herein by reference.
Such carboxy protecting groups are well known to those skilled in
the art, having been extensively used in the protection of carboxyl
groups in the penicillin and cephalosporin fields, as described in
U.S. Pat. Nos. 3,840,556 and 3,719,667, S. Kukolja, J. Am. Chem.
Soc. 93:6267-6269 (1971), and G. E. Gutowski, Tetrahedron Lett.
21:1779-1782 (1970), the disclosures of which are hereby
incorporated herein by reference. Examples of esters useful as
prodrugs for compounds containing carboxyl groups can be found, for
example, at pp. 14-21 in Bioreversible Carriers in Drug Design:
Theory and Application (E.B. Roche, ed.), Pergamon Press, New York
(1987), which is hereby incorporated herein by reference.
Representative carboxy protecting groups are C.sub.1 to C.sub.8
alkyl (e.g., methyl, ethyl or tertiary butyl and the like);
haloalkyl; alkenyl; cycloalkyl and substituted derivatives thereof
such as cyclohexyl, cyclopentyl and the like; cycloalkylalkyl and
substituted derivatives thereof such as cyclohexylmethyl,
cyclopentylmethyl and the like; arylalkyl, for example, phenethyl
or benzyl and substituted derivatives thereof such as alkoxybenzyl
or nitrobenzyl groups and the like; arylalkenyl, for example,
phenylethenyl and the like; aryl and substituted derivatives
thereof, for example, 5-indanyl and the like; dialkylaminoalkyl
(e.g., dimethylaminoethyl and the like); alkanoyloxyalkyl groups
such as acetoxymethyl, butyryloxymethyl, valerytoxymethyl,
isobutyryloxymethyl, isovaleryloxymethyl, 1-(propionyloxy)-1-ethyl,
1-(pivaloyloxyl)-1-ethyl, 1-methyl-1-(propionyloxy)-1-ethyl,
pivaloyloxymethyl, propionyloxymethyl and the like;
cycloalkanoyloxyalkyl groups such as cyclopropylcarbonyloxymethyl,
cyclobutylcarbonyloxymethyl, cyclopentylcarbonyloxymethyl,
cyclohexylcarbonyloxymethyl and the like; aroyloxyalkyl, such as
benzoyloxymethyl, benzoyloxyethyl and the like;
arylalkylcarbonyloxyalkyl, such as benzylcarbonyloxymethyl,
2-benzylcarbonyloxyethyl and the like; alkoxycarbonylalkyl, such as
methoxycarbonylmethyl, cyclohexyloxycarbonylmethyl,
1-methoxycarbonyl-1-ethyl, and the like; alkoxycarbonyloxyalkyl,
such as methoxycarbonyloxymethyl, t-butyloxycarbonyloxymethyl,
1-ethoxycarbonyloxy-1-ethyl, 1-cyclohexyloxycarbonyloxy-1-ethyl and
the like; alkoxycarbonylaminoalkyl, such as
t-butyloxycarbonylaminomethyl and the like;
alkylaminocarbonylaminoalkyl, such as
methylaminocarbonylaminomethyl and the like; alkanoylaminoalkyl,
such as acetylaminomethyl and the like;
heterocycliccarbonyloxyalkyl, such as
4-methylpiperazinylcarbonyloxymethyl and the like;
dialkylaminocarbonylalkyl, such as dimethylaminocarbonylmethyl,
diethylaminocarbonylmethyl and the like;
(5-(alkyl)-2-oxo-1,3-dioxolen-4-yl)alkyl, such as
(5-t-butyl-2-oxo-1,3-dioxolen-4-yl)methyl and the like; and
(5-phenyl-2-oxo-1,3-dioxolen-4-yl)alkyl, such as
(5-phenyl-2-oxo-1,3-dioxolen-4-yl)methyl and the like.
[0327] The term "N-protecting group" or "N-protected" as used
herein refers to those groups intended to protect the N-terminus of
an amino acid or peptide or to protect an amino group against
undesirable reactions during synthetic procedures. Commonly used
N-protecting groups are disclosed in, for example, Greene,
Protective Groups in Organic Synthesis, John Wiley & Sons, New
York (1981), which is hereby incorporated by reference. For
example, N-protecting groups can comprise acyl groups such as
formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl,
2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl,
o-nitrophenoxyacetyl, .alpha.-chlorobutyryl, benzoyl,
4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and the like;
sulfonyl groups such as benzenesulfonyl, p-toluenesulfonyl and the
like; carbamate forming groups such as benzyloxycarbonyl,
p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl,
p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl,
p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl,
3,5-dimethoxybenzyloxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl,
4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl,
3,4,5-trimethoxybenzyloxycarbonyl,
1-(p-biphenylyl)-1-methylethoxycarbonyl, .alpha.,.alpha.-di
methyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl,
t-butyloxycarbonyl, diisopropylmethoxycarbonyl,
isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl,
allyloxycarbonyl, 2,2,2,-trichloroethoxycarbonyl, phenoxycarbonyl,
4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl,
cyclopentyloxycarbonyl, adamantyloxycarbonyl,
cyclohexyloxycarbonyl, phenylthiocarbonyl and the like; alkyl
groups such as benzyl, triphenylmethyl, benzyloxymethyl and the
like; and silyl groups such as trimethylsilyl and the like. In some
embodiments N-protecting groups are formyl, acetyl, benzoyl,
pivaloyl, t-butylacetyl, phenylsulfonyl, benzyl,
9-fluorenylmethyloxycarbonyl (Fmoc), t-butyloxycarbonyl (Boc), and
benzyloxycarbonyl (Cbz).
[0328] As used herein, "halo," "halogen" or "halide" refers to F,
Cl, Br or I.
[0329] As used herein, the abbreviations for any protective groups,
amino acids or other compounds, are, unless indicated otherwise, in
accord with their common usage, recognized abbreviations, or the
IUPAC-IUB Commission on Biochemical Nomenclature, Biochem.
11:942-944 (1972).
[0330] As used herein, "substantially pure" means sufficiently
homogeneous to appear free of readily detectable impurities as
determined by standard methods of analysis, such as thin layer
chromatography (TLC), gel electrophoresis, and high performance
liquid chromatography (HPLC), used by those of skill in the art to
assess such purity, or sufficiently pure such that further
purification would not detectably alter the physical and chemical
properties, such as enzymatic and biological activities, of the
substance. Substantially pure includes compositions in which the AA
targeting agent or AA targeting compound forms the major component
of the composition, such as constituting about 50%, about 60%,
about 70%, about 80%, about 90%, or about 95% or more of the
substances in the composition. Methods for purification of
compounds to produce substantially chemically pure compounds are
known to those of skill in the art. A substantially chemically pure
compound may, however, be a mixture of stereoisomers. In such
instances, further purification may increase the specific activity
of the compound. However, AA targeting agents need not always be
provided in a specific purified state. Partially purified
compositions will have utility in certain embodiments and depending
on the desired use. For example, purification methods that may
yield a greater total recovery of AA-targeting agent may produce a
lower degree of relative purification.
[0331] As used herein, "biological activity" refers to the in vivo
activities of a compound, composition, or other mixture, or
physiological responses that result upon in vivo administration of
a compound, composition or other mixture. Biological activity thus
encompasses therapeutic effects, diagnostic effects and
pharmaceutical activity of such compounds, compositions, and
mixtures. The term "biologically active" or "functional" when used
as a modifier of invention AA targeting agent containing
polypeptides or compositions thereof refers to a polypeptide that
exhibits at least one activity that is characteristic of or similar
to an AA targeting agent.
[0332] As used herein, "pharmacokinetics" refers to the
concentration of an administered compound in the serum over time.
Pharmacodynamics refers to the concentration of an administered
compound in target and nontarget tissues over time and the effects
on the target tissue (e.g., efficacy) and the non-target tissue
(e.g., toxicity). Improvements in, for example, pharmacokinetics or
pharmacodynamics can be designed for a particular targeting agent
or biological agent, such as by using labile linkages or by
modifying the chemical nature of any linker (e.g., changing
solubility, charge, and the like).
[0333] As employed herein, the phrases "an effective amount" and
"therapeutically effective amount" refer to an amount of an AA
targeting agent or compound comprising an AA targeting agent that
is useful or able to support an observable change in the level of
one or more biological activity characteristic of an AA targeting
agent, or a dose sufficient to impart a beneficial effect, e.g., an
amelioration of a symptom on the recipient thereof. The specific
therapeutically effective dose level for any particular subject
will depend upon a variety of factors including the symptom or
disorder being treated, the severity of the symptom or disorder,
the activity of the specific compound, the route of administration,
the rate of clearance of the compound, the duration of treatment,
the drugs used in combination or coincident with the compound, the
age, body weight, sex, diet, and general health of the subject, and
like, as well as other factors well known in the medical arts and
sciences. A therapeutically effective amount can be an amount of AA
targeting compound sufficient to produce a measurable inhibition of
angiogenesis in the tissue being treated, i.e., an
angiogenesis-inhibiting amount. Inhibition of angiogenesis can be
measured in situ by immunohistochemistry, or by other methods known
to one skilled in the art. Various general considerations taken
into account in determining the "therapeutically effective amount"
are known to those of skill in the art and are described, e.g., in
Gilman, A. G., et al., Goodman And Gilman's The Pharmacological
Basis of Therapeutics, 8.sup.th ed., McGraw-Hill (1990); and
Remington's Pharmaceutical Sciences, 17.sup.th ed., Mack Publishing
Co., Easton, Pa. (1990).
[0334] In one aspect, the present invention provides various
targeting compounds in which AA targeting agents are covalently
linked to a combining site of an antibody.
[0335] In another aspect, the present invention includes methods of
altering at least one physical or biological characteristic of an
AA targeting agent. The methods include covalently linking an AA
targeting agent to a combining site of an antibody, either directly
or though a linker. Characteristics of an AA targeting agent that
may be modified include, but are not limited to, binding affinity,
susceptibility to degradation (e.g., by proteases),
pharmacokinetics, pharmacodynamics, immunogenicity, solubility,
lipophilicity, hydrophilicity, hydrophobicity, stability (either
more or less stable, as well as planned degradation), rigidity,
flexibility, modulation of antibody binding, and the like. Also,
the biological potency of a particular AA targeting agent may be
increased by the addition of the effector function(s) provided by
the antibody. For example, an antibody provides effector functions
such as complement mediated effector functions. Without wishing to
be bound by any theory, the antibody portion of an AA targeting
compound may generally extend the half-life of a smaller sized AA
targeting agent in vivo. Thus, in one aspect, the invention
provides a method for increasing the effective circulating
half-life of an AA targeting agent.
[0336] In another aspect, the present invention provides methods
for modulating the binding activity of an antibody by covalently
attaching an AA targeting agent to a combining site of the
antibody. Although not wishing to be bound by any theory,
substantially reduced antibody binding to an antigen may result
from the linked AA targeting agent(s) sterically hindering the
antigen from contacting the antibody combining site. Alternatively,
substantially reduced antigen binding may result if the amino acid
side chain of the antibody combining site modified by covalent
linkage is important for binding to the antigen. By contrast,
substantially increased antibody binding to an antigen may result
when a linked AA targeting agent(s) does not sterically hinder the
antigen from contacting the antibody combining site and/or when the
amino acid side chain of the antibody combining site modified by
covalent linkage is not important for binding to the antigen.
[0337] In another aspect, the present invention includes methods of
modifying a combining site of an antibody to generate binding
specificity for Ang-2. Such methods include covalently linking a
reactive amino acid side chain in a combining site of the antibody
to a chemical moiety on a linker of an AA targeting agent-linker
compound as described herein where an AA targeting agent is based
upon an Ang-2 binding peptide. The chemical moiety of the linker is
sufficiently distanced from the AA targeting agent so that an AA
targeting agent can bind its cognate when an AA targeting
agent-linker compound is covalently linked to an antibody combining
site. Typically, the antibody will not be considered specific for
the target molecule. In certain embodiments, an antibody prior to
covalent linking would have an affinity for Ang-2 of less than
about 1.times.10.sup.-5 moles/liter. However, after the antibody is
covalently linked to the AA targeting agent-linker compound, the
modified antibody preferably has an affinity for the target
molecule of at least about 1.times.10.sup.-6 moles/liter,
alternatively, at least about 1.times.10.sup.-7 moles/liter,
alternatively, at least 1.times.10.sup.-8 moles/liter,
alternatively at least 1.times.10.sup.-9 moles/liter, or
alternatively, at least about 1.times.10.sup.-10 moles/liter.
AA Targeting Agents
[0338] An AA targeting agent is a peptide selected from the group
consisting of: [0339] R.sup.1-Q(AcK)Y QPL DEL DKT LYD QFM LQQ
G-R.sup.2; (SEQ ID NO:29) [0340] R.sup.1-QNY QPL DEL DKT LYD QFM
LQQ G-R.sup.2; (SEQ ID NO:28) [0341] R.sup.1-Q(AcK)Y QPL DEK
D(AcK)T LYD QFM LQQ G-R.sup.2; (SEQ ID NO:30) [0342]
R.sup.1-Q(AcK)Y QPL DEK DET LYD QFM LQQ G-R.sup.2; (SEQ ID NO:33)
[0343] R.sup.1-Q(AcK)Y Q(DHP)L DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ
ID NO:34) [0344] R.sup.1-Q(AcK)Y Q(HP)L DEL DKT LYD QFM LQQ
G-R.sup.2; (SEQ ID NO:35) [0345] R.sup.1-Q(AcK)Y QPL DEL DKT IYD
QFM LQQ G-R.sup.2; (SEQ ID NO:36) [0346] R.sup.1-Q(AcK)Y QPL DEI
DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:37) [0347] R.sup.1-Q(AcK)Y
QPL DEL DKT (HChA)YD QFM LQQ G-R.sup.2; (SEQ ID NO:38) [0348]
R.sup.1-Q(AcK)Y QPL DEL DKT (HF)YD QFM LQQ G-R.sup.2; (SEQ ID
NO:39) [0349] R.sup.1-Q(AcK)Y QPL DEL DKT (ThA)YD QFM LQQ
G-R.sup.2; (SEQ ID NO:40) [0350] R.sup.1-Q(AcK)Y QPL DEL DKT
(Nva)YD QFM LQQ G-R.sup.2; (SEQ ID NO:41) [0351] R.sup.1-Q(AcK)Y
QPL DEL DKT (HL)YD QFM LQQ G-R.sup.2; (SEQ ID NO:42) [0352]
R.sup.1-Q(AcK)Y QPL DE(AcK) DKT LYD QFM LQQ G-R.sup.2; (SEQ ID
NO:43) [0353] R.sup.1-Q(AcK)Y QPL DEL DKT LFD QFM LQQ G-R.sup.2;
(SEQ ID NO:44) [0354] R.sup.1-Q(AcK)Y Q(HP)L DE(ThA) DKT L(NO2F)D
QFM LQQ G-R.sup.2; (SEQ ID NO:45) [0355] R.sup.1-Q(AcK)Y Q(HP)L
DE(ThA) DKT L(BPA)D QFM LQQ G-R.sup.2; (SEQ ID NO:46) [0356]
R.sup.1-Q(AcK)Y Q(HP)L DE(ThA) DKT L(CO2H)FD QFM LQQ G-R.sup.2;
(SEQ ID NO:47) [0357] R.sup.1-Q(AcK)Y QPL DE(AcK) DKT L(NO2F)D QFM
LQQ G-R.sup.2; (SEQ ID NO:48) [0358] R.sup.1-Q(AcK)Y QPL DE(AcK)
DKT LFD QFM LQQ G-R.sup.2 (SEQ ID NO:63) [0359] DCB-Q(AcK)Y QPL DEL
DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:49) [0360] DFB-Q(AcK)Y QPL
DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:50) [0361] PyC-Q(AcK)Y
QPL DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:51) [0362] Amido
2-PEG-Q(AcK)Y QPL DEL DKT LYD QFM LQQ G-R.sup.2; (SEQ ID NO:52)
[0363] R.sup.1-Q(CIBnCarbamateK)Y QPL DEL DKT LYD QFM LQQ
G-R.sup.2; (SEQ ID NO:53) [0364] R.sup.1-Q(AcK)Y QPL DEL D(Dab)T
LYD QFM LQQ G-R.sup.2; (SEQ ID NO:54) [0365] R.sup.1-Q(AcK)Y QPL
DEL D(Dap)T LYD QFM LQQ G-R.sup.2; (SEQ ID NO:55) [0366]
R.sup.1-Q(AcK)Y QPL DEL DKT L(NO2F)D QFM LQQ G-R.sup.2; (SEQ ID
NO:91) [0367] R.sup.1-QNY QPL DEL DKT L(BPA)D QFM LQQ G-R.sup.2;
(SEQ ID NO:56) [0368] R.sup.1-QNY QPL DEL DKT L(CF)D QFM LQQ
G-R.sup.2; (SEQ ID NO:57) [0369] R.sup.1-Q(Nick)Y QPL DEL DKT LYD
QFM LQQ G-R.sup.2; (SEQ ID NO:61) [0370] R.sup.1-Q(AcK)Y QPL
DE(AcK) DKT L(CF)D QFM LQQ G-R.sup.2; (SEQ ID NO:62) and [0371]
R.sup.1-Q(AcK)Y QPL DE(AcK) DKT LFD QFM LQQ G-R.sup.2; (SEQ ID NO:
63) [0372] wherein [0373] R.sup.1 is CH.sub.3, C(O)CH.sub.3,
C(O)CH.sub.3, C(O)CH.sub.2CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.3, C(O)CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
C(O)CH(CH.sub.3)CH.sub.2CH.sub.3, C(O)C.sub.6H.sub.5,
C(O)CH.sub.2CH.sub.2(CH.sub.2CH.sub.2O).sub.1-5Me, dichlorobenzoyl
(DCB), difluorobenzoyl (DFB), pyridinyl carboxlate (PyC) or
amido-2-PEG, an amino protecting group, a lipid fatty acid group or
a carbohydrate; and [0374] R.sup.2 is OH, NH.sub.2, NH(CH.sub.3),
NHCH.sub.2CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.3, NHCH.sub.2CH.sub.2CH.sub.2CH.sub.3,
NHCH(CH.sub.3)CH.sub.2CH.sub.3, NHC.sub.6H.sub.5,
NHCH.sub.2CH.sub.2OCH.sub.3, NHOCH.sub.3, NHOCH.sub.2CH.sub.3, a
carboxy protecting group, a lipid fatty acid group or a
carbohydrate, and
[0375] An AA targeting compound can be prepared using techniques
well known in the art. Typically, synthesis of the peptidyl AA
targeting agent is the first step and is carried out as described
herein. The targeting agent is then derivatized for linkage to a
connecting component (the linker), which is then combined with the
antibody. One of skill in the art will readily appreciate that the
specific synthetic steps used depend upon the exact nature of the
three components. Thus, AA targeting agent-linker conjugates and AA
targeting compounds described herein can be readily
synthesized.
[0376] AA targeting agent peptides may be synthesized by many
techniques that are known to those skilled in the art. For solid
phase peptide synthesis, a summary of exemplary techniques may be
found in Chemical Approaches to the Synthesis of Peptides and
Proteins (Williams et al., eds.), CRC Press, Boca Raton, Fla.
(1997).
[0377] Typically, the desired peptidic AA targeting agent is
synthesized sequentially on solid phase according to procedures
well known in the art. See, e.g., U.S. Patent Application No.
2003/0045477). The linker may be attached to the peptide in part or
in full on the solid phase, or may be added using solution phase
techniques after the removal of the peptide from the resin (see
FIGS. 6A and 6B). For example, an N-protected amino and carboxylic
acid-containing linking moiety may be attached to a resin such as
4-hydroxymethyl-phenoxymethyl-poly(styrene-1% divinylbenzene). The
N-protecting group may be removed by the appropriate acid (e.g.,
TFA for Boc) or base (e.g., piperidine for Fmoc), and the peptide
sequence developed in the normal C-terminus to N-terminus fashion
(see FIG. 6A). Alternatively, the peptide sequence may be
synthesized first and the linker added to the N-terminal amino acid
residue last (see FIG. 6B). Yet another method entails deprotecting
an appropriate side chain during synthesis and derivatizing with a
suitably reactive linker. For example, a lysine side chain may be
deprotected and reacted with a linker having an active ester.
Alternatively, an amino acid derivative with a suitably protected
linker moiety already attached to the side chain (see FIG. 6B) or,
in some cases, the alpha-amino nitrogen, may be added as part of
the growing peptide sequence.
[0378] At the end of the solid phase synthesis, the targeting
agent-linker conjugate is removed from the resin and deprotected,
either in succession or in a single operation. Removal of the
targeting agent-linker conjugate and deprotection can be
accomplished in a single operation by treating the resin-bound
peptide-linker conjugate with a cleavage reagent, for example,
trifluoroacetic acid containing scavengers such as thianisole,
water, or ethanedithiol. After deprotection and release of the
targeting agent, further derivatization of the targeting agent
peptide may be carried out.
[0379] The fully deprotected targeting agent-linker conjugate is
purified by a sequence of chromatographic steps employing any or
all of the following types: ion exchange on a weakly basic resin in
the acetate form; hydrophobic adsorption chromatography on
underivatized polystyrene-divinylbenzene (e.g., AMBERLITE XAD);
silica gel adsorption chromatography; ion exchange chromatography
on carboxymethylcellulose; partition chromatography, e.g., on
SEPHADEX G-25, LH-20 or countercurrent distribution; high
performance liquid chromatography (HPLC), especially reverse-phase
HPLC on octyl- or octadecylsilyl-silica bonded phase column
packing.
Antibodies
[0380] "Antibody" as used herein includes polypeptide molecules
comprising heavy and/or light chains which have immunoreactive
activity. Antibodies include immunoglobulins which are the product
of B cells and variants thereof, as well as the T cell receptor
(TcR) which is the product of T cells and variants thereof. An
immunoglobulin is a protein comprising one or more polypeptides
substantially encoded by the immunoglobulin kappa and lambda,
alpha, gamma, delta, epsilon and mu constant region genes, as well
as myriad immunoglobulin variable region genes. Light chains are
classified as either kappa or lambda. Heavy chains are classified
as gamma, mu, alpha, delta, or epsilon, which in turn define the
immunoglobulin classes, IgG, IgM, IgA, IgD, and IgE, respectively.
Subclasses of heavy chains are also known. For example, IgG heavy
chains in humans can be any of IgG1, IgG2, IgG3, and IgG4
subclasses.
[0381] A typical immunoglobulin structural unit is known to
comprise a tetramer. Each tetramer is composed of two identical
pairs of polypeptide chains, each pair having one "light" (about 25
kD) and one "heavy" chain (about 50-70 kD). The N-terminus of each
chain defines a variable region of about 100 to 110 or more amino
acids primarily responsible for antigen recognition. The terms
variable light chain (V.sub.L) and variable heavy chain (V.sub.H)
refer to these light and heavy chains respectively. The amino acids
of an antibody may be naturally or nonnaturally occurring.
[0382] Antibodies that contain two combining sites are bivalent in
that they have two complementarity or antigen recognition sites. A
typical natural bivalent antibody is an IgG. Although vertebrate
antibodies generally comprise two heavy chains and two light
chains, heavy chain only antibodies are also known. See Muyldermans
et al., TRENDS in Biochem. Sci. 26(4):230-235 (1991). Such
antibodies are bivalent and are formed by the pairing of heavy
chains. Antibodies may also be multi-valent, as in the case of
dimeric forms of IgA and the pentameric IgM molecule. Antibodies
also include hybrid antibodies wherein the antibody chains are
separately homologous with referenced mammalian antibody chains.
One pair of heavy and light chain has a combining site specific to
one antigen and the other pair of heavy and light chains has a
combining site specific to a different antigen. Such antibodies are
referred to as bi-specific because they are able to bind two
different antigens at the same time. Antibodies may also be
univalent, such as, for example, in the case of Fab or Fab'
fragments.
[0383] Antibodies exist as full length intact antibodies or as a
number of well-characterized fragments produced by digestion with
various peptidases or chemicals. Thus, for example, pepsin digests
an antibody below the disulfide linkages in the hinge region to
produce F(ab').sub.2, a dimer of Fab which itself is a light chain
joined to V.sub.H--CH.sub.1 by a disulfide bond. F(ab').sub.2 may
be reduced under mild conditions to break the disulfide linkage in
the hinge region, thereby converting the F(ab').sub.2 dimer into a
Fab' monomer. The Fab' monomer is essentially a Fab fragment with
part of the hinge region (see, e.g., Fundamental Immunology (W. E.
Paul, ed.), Raven Press, N.Y. (1993) for a more detailed
description of other antibody fragments). As another example,
partial digestion with papain can yield a monovalent Fab/c
fragment. See M. J. Glennie et al., Nature 295:712-714 (1982).
While various antibody fragments are defined in terms of the
digestion of an intact antibody, one of skill in the art will
appreciate that any of a variety of antibody fragments may be
synthesized de novo either chemically or by utilizing recombinant
DNA methodology. Thus, the term antibody as used herein also
includes antibody fragments produced by the modification of whole
antibodies, synthesized de novo, or obtained from recombinant DNA
methodologies. One skilled in the art will recognize that there are
circumstances in which it is advantageous to use antibody fragments
rather than whole antibodies. For example, the smaller size of the
antibody fragments allows for rapid clearance and may lead to
improved access to solid tumors. Recombinant antibodies may be
conventional full length antibodies, hybrid antibodies, heavy chain
antibodies, antibody fragments known from proteolytic digestion,
antibody fragments such as Fv or single chain Fv (scFv), single
domain fragments such as V.sub.H or V.sub.L, diabodies, domain
deleted antibodies, minibodies, and the like. An Fv antibody is
about 50 kD in size and comprises the variable regions of the light
and heavy chain. The light and heavy chains may be expressed in
bacteria where they assemble into an Fv fragment. Alternatively,
the two chains can be engineered to form an interchain disulfide
bond to give a dsFv. A single chain Fv ("scFv") is a single
polypeptide comprising V.sub.H and V.sub.L sequence domains linked
by an intervening linker sequence, such that when the polypeptide
folds the resulting tertiary structure mimics the structure of the
antigen binding site. See J. S. Huston et al., Proc. Nat. Acad.
Sci. U.S.A. 85:5879-5883 (1988). One skilled in the art will
recognize that depending on the particular expression method and/or
antibody molecule desired, appropriate processing of the
recombinant antibodies may be performed to obtain a desired
reconstituted or reassembled antibody. See, e.g., Vallejo and
Rinas, Biomed Central., available at world wide web URL
microbialcellfactories.com/content/3/1/11.
[0384] Single domain antibodies are the smallest functional binding
units of antibodies (approximately 13 kD in size), corresponding to
the variable regions of either the heavy V.sub.H or light V.sub.L
chains. See U.S. Pat. No. 6,696,245, WO04/058821, WO04/003019 and
WO03/002609. Single domain antibodies are well expressed in
bacteria, yeast, and other lower eukaryotic expression systems.
Domain deleted antibodies have a domain, such as CH2, deleted
relative to the full length antibody. In many cases such domain
deleted antibodies, particularly CH2 deleted antibodies, offer
improved clearance relative to their full length counterparts.
Diabodies are formed by the association of a first fusion protein
comprising two V.sub.H domains with a second fusion protein
comprising two V.sub.L domains. Diabodies, like full length
antibodies, are bivalent and may be bi-specific. Minibodies are
fusion proteins comprising a V.sub.H, V.sub.L, or scFv linked to
CH3, either directly or via an intervening IgG hinge. See T.
Olafsen et al., Protein Eng. Des. Sel. 17:315-323 (2004).
Minibodies, like domain deleted antibodies, are engineered to
preserve the binding specificity of full-length antibodies but with
improved clearance due to their smaller molecular weight.
[0385] The T cell receptor (TcR) is a disulfide linked heterodimer
composed of two chains. The two chains are generally
disulfide-bonded just outside the T cell plasma membrane in a short
extended stretch of amino acids resembling the antibody hinge
region. Each TcR chain is composed of one antibody-like variable
domain and one constant domain. The full TcR has a molecular mass
of about 95 kD, with the individual chains varying in size from 35
to 47 kD. Also encompassed within the meaning of TcR are portions
of the receptor, such as, for example, the variable region, which
can be produced as a soluble protein using methods well known in
the art. For example, U.S. Pat. No. 6,080,840 and A. E. Slanetz and
A. L. Bothwell, Eur. J. Immunol. 21:179-183 (1991) describe a
soluble T cell receptor prepared by splicing the extracellular
domains of a TcR to the glycosyl phosphatidylinositol (GPI)
membrane anchor sequences of Thy-1. The molecule is expressed in
the absence of CD3 on the cell surface, and can be cleaved from the
membrane by treatment with phosphatidylinositol specific
phospholipase C (PI-PLC). The soluble TcR also may be prepared by
coupling the TcR variable domains to an antibody heavy chain
CH.sub.2 or CH.sub.3 domain, essentially as described in U.S. Pat.
No. 5,216,132 and G. S. Basi et al., J. Immunol. Methods
155:175-191 (1992), or as soluble TcR single chains, as described
by E. V. Shusta et al., Nat. Biotechnol. 18:754-759 (2000) or P. D.
Holler et al., Proc. Natl. Acad. Sci. U.S.A. 97:5387-5392 (2000).
Certain embodiments of the invention use TcR "antibodies" as a
soluble antibody. The combining site of the TcR can be identified
by reference to CDR regions and other framework residues using the
same methods discussed above for antibodies.
[0386] The combining site refers to the part of an antibody
molecule that participates in antigen binding. The antigen binding
site is formed by amino acid residues of the N-terminal variable
("V") regions of the heavy ("H") and light ("L") chains. The
antibody variable regions comprise three highly divergent stretches
referred to as "hypervariable regions" or "complementarity
determining regions" (CDRs), which are interposed between more
conserved flanking stretches known as "framework regions" (FRs).
The three hypervariable regions of a light chain (LCDR1, LCDR2, and
LCDR3) and the three hypervariable regions of a heavy chain (HCDR1,
HCDR2, and HCDR3) are disposed relative to each other in three
dimensional space to form an antigen binding surface or pocket. In
heavy-chain antibodies or V.sub.H domains, the antigen binding site
is formed by the three hypervariable regions of the heavy chains.
In V.sub.L domains, the antigen binding site is formed by the three
hypervariable regions of the light chain.
[0387] The identity of the amino acid residues in a particular
antibody that make up a combining site can be determined using
methods well known in the art. For example, antibody CDRs may be
identified as the hypervariable regions originally defined by Kabat
et al. See E. A. Kabat et al., Sequences of Proteins of
Immunological Interest, 5.sup.th ed., Public Health Service, NIH,
Washington D.C. (1992). The positions of the CDRs may also be
identified as the structural loop structures originally described
by Chothia and others. See, e.g., C. Chothia and A. M. Lesk, J.
Mol. Biol. 196:901-917 (1987); C. Chothia et al., Nature
342:877-883 (1989); and A. Tramontano et al., J. Mol. Biol.
215:175-182 (1990). Other methods include the "AbM definition,"
which is a compromise between Kabat and Chothia and is derived
using Oxford Molecular's AbM antibody modeling software (now
Accelrys), or the "contact definition" of CDRs set forth in R. M.
MacCallum et al., J. Mol. Biol. 262:732-745 (1996). Table 2
identifies CDRs based upon various known definitions:
TABLE-US-00021 TABLE 2 CDR definitions CDR Kabat AbM Chothia
Contact L1 L24-L34 L24-L34 L24-L34 L30-L36 L2 L50-L56 L50-L56
L50-L56 L46-L55 L3 L89-L97 L89-L97 L89-L97 L89-L96 H1 (Kabat
H31-H35B H26- H26-H32 . . . H34 H30- numbering) H35B H35B H1
(Chothia H31-H35 H26-H35 H26-H32 H30-H35 numbering) H2 H50-H56
H50-H58 H52-H56 H47-H58 H3 H95-H102 H95-H102 H95-H102 H93-H101
[0388] General guidelines by which one may identify the CDRs in an
antibody from sequence alone are as follows: [0389] LCDR1:
[0390] Start--Approximately residue 24.
[0391] Residue before is always a Cys.
[0392] Residue after is always a Trp, typically followed by
Tyr-Gln, but also followed by Leu-Gln, Phe-Gln, or Tyr-Leu.
[0393] Length is 10 to 17 residues. [0394] LCDR2:
[0395] Start--16 residues after the end of L1.
[0396] Sequence before is generally Ile-Tyr, but also may be
Val-Tyr, Ile-Lys, or Ile-Phe.
[0397] Length is generally 7 residues. [0398] LCDR3:
[0399] Start--33 residues after end of L2.
[0400] Residue before is a Cys.
[0401] Sequence after is Phe-Gly-X-Gly.
[0402] Length is 7 to 11 residues. [0403] HCDR1:
[0404] Start--approximately residue 26, four residues after a Cys
under Chothia/AbM definitions; start is 5 residues later under
Kabat definition.
[0405] Sequence before is Cys-X--X--X.
[0406] Residue after is a Trp, typically followed by Val, but also
followed by Ile or Ala.
[0407] Length is 10 to 12 residues under AbM definition; Chothia
definition excludes the last 4 residues. [0408] HCDR2:
[0409] Start--15 residues after the end of Kabat/AbM definition of
CDR-H1.
[0410] Sequence before is typically Leu-Glu-Trp-Ile-Gly, but a
number of variations are possible.
[0411] Sequence after is
Lys/Arg-Leu/Ile/Val/Phe/Thr/Ala-Thr/Ser/Ile/Ala.
[0412] Length is 16 to 19 residues under Kabat definition; AbM
definition excludes the last 7 residues. [0413] HCDR3:
[0414] Start--33 residues after end of CDR-H2 (two residues after a
Cys).
[0415] Sequence before is Cys-X--X (typically Cys-Ala-Arg).
[0416] Sequence after is Trp-Gly-X-Gly.
[0417] Length is 3 to 25 residues.
[0418] The identity of the amino acid residues in a particular
antibody that are outside the CDRs, but nonetheless make up part of
the combining site by having a side chain that is part of the
lining of the combining site (i.e., that is available to linkage
through the combining site), can be determined using methods well
known in the art, such as molecular modeling and X-ray
crystallography. See, e.g., L. Riechmann et al., Nature 332:323-327
(1988).
[0419] As discussed, antibodies that can be used in preparing
antibody-based AA targeting compounds require a reactive side chain
in the antibody combining site. A reactive side chain may be
present naturally or may be placed in an antibody by mutation. The
reactive residue of the antibody combining site may be associated
with the antibody, such as when the residue is encoded by nucleic
acid present in the lymphoid cell first identified to make the
antibody. Alternatively, the amino acid residue may arise by
purposely mutating the DNA so as to encode the particular residue
(see, e.g., WO 01/22922 to Meares et al.). The reactive residue may
be a non-natural residue arising, for example, by biosynthetic
incorporation using a unique codon, tRNA, and aminoacyl-tRNA as
discussed herein. In another approach, the amino acid residue or
its reactive functional groups (e.g., a nucleophilic amino group or
sulfhydryl group) may be attached to an amino acid residue in the
antibody combining site. Thus, covalent linkage with the antibody
occurring "through an amino acid residue in a combining site of an
antibody" as used herein means that linkage can be directly to an
amino acid residue of an antibody combining site or through a
chemical moiety that is linked to a side chain of an amino acid
residue of an antibody combining site.
[0420] Catalytic antibodies are one source of antibodies with
combining sites that comprise one or more reactive amino acid side
chains. Such antibodies include aldolase antibodies, beta lactamase
antibodies, esterase antibodies, amidase antibodies, and the
like.
[0421] One embodiment comprises an aldolase antibody such as the
mouse monoclonal antibody mAb 38C2 or mAb 33F12, as well as
suitably humanized and chimeric versions of such antibodies. Mouse
mAb 38C2 has a reactive lysine near to but outside HCDR3, and is
the prototype of a new class of catalytic antibodies that were
generated by reactive immunization and mechanistically mimic
natural aldolase enzymes. See C. F. Barbas 3.sup.rd et al., Science
278:2085-2092 (1997)). Other aldolase catalytic antibodies that may
be used include the antibodies produced by the hybridoma 85A2,
having ATCC accession number PTA-1015; hybridoma 85C7, having ATCC
accession number PTA-1014; hybridoma 92F9, having ATCC accession
number PTA-1017; hybridoma 93F3, having ATCC accession number
PTA-823; hybridoma 84G3, having ATCC accession number PTA-824;
hybridoma 84G11, having ATCC accession number PTA-1018; hybridoma
84H9, having ATCC accession number PTA-1019; hybridoma 85H6, having
ATCC accession number PTA-825; hybridoma 90G8, having ATCC
accession number PTA-1016. Through a reactive lysine, these
antibodies catalyze aldol and retro-aldol reactions using the
enamine mechanism of natural aldolases. See, e.g., J. Wagner et
al., Science 270:1797-1800 (1995); C. F. Barbas 3.sup.rd et al.,
Science 278:2085-2092 (1997); G. Zhong et al., Angew. Chem. Int.
Ed. Engl. 38:3738-3741 (1999); A. Karlstrom et al., Proc. Natl.
Acad. Sci. U.S.A., 97:3878-3883 (2000). Aldolase antibodies and
methods of generating aldolase antibodies are disclosed in U.S.
Pat. Nos. 6,210,938, 6,368,839, 6,326,176, 6,589,766, 5,985,626,
and 5,733,757.
[0422] AA targeting compounds may also be formed by linking an AA
targeting agent to a reactive cysteine, such as those found in the
combining sites of thioesterase and esterase catalytic antibodies.
Suitable thioesterase catalytic antibodies are described by K. D.
Janda et al., Proc. Natl. Acad. Sci. U.S.A. 91:2532-2536 (1994).
Suitable esterase antibodies are described by P. Wirsching et al.,
Science 270:1775-1782 (1995). Reactive amino acid-containing
antibodies may be prepared by means well known in the art,
including mutating an antibody combining site residue to encode for
the reactive amino acid or chemically derivatizing an amino acid
side chain in an antibody combining site with a linker that
contains the reactive group.
[0423] Antibodies suitable for use herein may be obtained by
conventional immunization, reactive immunization in vivo, or by
reactive selection in vitro, such as with phage display. Antibodies
may also be obtained by hybridoma or cell fusion methods or in
vitro host cells expression system. Antibodies may be produced in
humans or in other animal species. Antibodies from one species of
animal may be modified to reflect another species of animal. For
example, human chimeric antibodies are those in which at least one
region of the antibody is from a human immunoglobulin. A human
chimeric antibody is typically understood to have variable region
amino acid sequences homologous to a non-human animal, e.g., a
rodent, with the constant region having amino acid sequence
homologous to a human immunoglobulin. In contrast, a humanized
antibody uses CDR sequences from a non-human antibody with most or
all of the variable framework region sequence and all the constant
region sequence from a human immunoglobulin. Chimeric and humanized
antibodies may be prepared by methods well known in the art
including CDR grafting approaches (see, e.g., N. Hardman et al.,
Int. J. Cancer 44:424-433 (1989); C. Queen et al., Proc. Natl.
Acad. Sci. U.S.A. 86:10029-10033 (1989)), chain shuffling
strategies (see, e.g., Rader et al., Proc. Natl. Acad. Sci. U.S.A.
95:8910-8915 (1998), genetic engineering molecular modeling
strategies (see, e.g., M. A. Roguska et al., Proc. Natl. Acad. Sci.
U.S.A. 91:969-973 (1994)), and the like.
[0424] Methods for humanizing non-human antibodies have been
described in the art. Preferably, a humanized antibody has one or
more amino acid residues introduced into it from a source which is
non-human. These non-human amino acid residues are often referred
to as "import" residues, which are typically taken from an "import"
variable domain. Humanization can be essentially performed
following the methods of Winter and colleagues (see, e.g., P. T.
Jones et al., Nature 321:522-525 (1986); L. Riechmann et al.,
Nature 332:323-327 (1988); M. Verhoeyen et al., Science
239:1534-1536 (1988)) by substituting hypervariable region
sequences for the corresponding sequences of a human antibody.
Accordingly, such "humanized" antibodies are chimeric antibodies
wherein substantially less than an intact human variable domain has
been substituted by the corresponding sequence from a non-human
species. In practice, humanized antibodies are typically human
antibodies in which some hypervariable region residues and possibly
some framework (FR) residues are substituted by residues from
analogous sites in rodent antibodies.
[0425] The choice of human variable domains, both light and heavy,
to be used in making humanized antibodies is very important to
reduce antigenicity and human anti-mouse antibody (HAMA) response
when the antibody is intended for human therapeutic use. According
to the so-called "best-fit" method, the human variable domain
utilized for humanization is selected from a library of known
domains based on a high degree of homology with the rodent variable
region of interest (M. J. Sims et al., J. Immunol., 151:2296-2308
(1993); M. Chothia and A. M. Lesk, J. Mol. Biol. 196:901-917
(1987)). Another method uses a framework region derived from the
consensus sequence of all human antibodies of a particular subgroup
of light or heavy chains. The same framework may be used for
several different humanized antibodies (see, e.g., P. Carter et
al., Proc. Natl. Acad. Sci. U.S.A. 89:4285-4289 (1992); L. G.
Presta et al., J. Immunol., 151:2623-2632 (1993)).
[0426] It is further important that antibodies be humanized with
retention of high linking affinity for the Z group. To achieve this
goal, according to one method, humanized antibodies are prepared by
analysis of the parental sequences and various conceptual humanized
products using three-dimensional models of the parental and
humanized sequences. Three-dimensional immunoglobulin models are
commonly available and are familiar to those skilled in the art.
Computer programs are available which illustrate and display
probable three-dimensional conformational structures of selected
candidate immunoglobulin sequences. Inspection of these displays
permits analysis of the likely role of the residues in the
functioning of the candidate immunoglobulin sequence with respect
to linking to the Z group. In this way, FR residues can be selected
and combined from the recipient and import sequences so that the
desired antibody characteristic, such as increased affinity for the
target antigen(s), is achieved.
[0427] Various forms of humanized murine aldolase antibodies are
contemplated. One embodiment uses the humanized aldolase catalytic
antibody h38c2 IgG1 or h38c2 Fab with human constant domains C and
C.sub.11. C. Rader et al., J. Mol. Bio. 332:889-899 (2003)
discloses the gene sequences and vectors that may be used to
produce h38c2 Fab and h38c2 IgG1. Human germline V.sub.k gene DPK-9
and human J.sub.k gene JK4 were used as frameworks for the
humanization of the kappa light chain variable domain of m38c2, and
human germline gene DP-47 and human J.sub.H gene JH4 were used as
frameworks for the humanization of the heavy chain variable domain
of m38c2. FIG. 7A illustrates a sequence alignment between the
variable light and heavy chains in m38c2, h38c2, and human
germlines. h38c2 may utilize IgG1, IgG2, IgG3, or IgG4 constant
domains, including any of the allotypes thereof. FIG. 7B
illustrates one embodiment of h38c2 IgG1 using the G1 m(f)
allotype, where the light and heavy chain amino acid sequences of
this h38c2 IgG1 are set forth in the figure. In certain embodiments
of AA targeting compounds of formula II or III wherein Antibody is
h38c2 IgG1 with the G1 m(f) allotype, Z binds to the side chain of
the lysine residue at position 99 of the heavy chain. This residue
is denoted by bold print in FIG. 7B. Another embodiment uses a
chimeric antibody comprising the variable domains (V.sub.L and
V.sub.H) of h38c2 and the constant domains from an IgG1, IgG2,
IgG3, or IgG4.
[0428] Various forms of humanized aldolase antibody fragments are
also contemplated. One embodiment uses h38c2 F(ab').sub.2. h38c2
F(ab').sub.2 may be produced by the proteolytic digestion of h38c2
IgG1. Another embodiment uses an h38c2 scFv comprising the V.sub.L
and V.sub.H domains from h38c2 which are optionally connected by
the intervening linker (Gly.sub.4Ser).sub.3.
[0429] As an alternative to humanization, human antibodies can be
generated. For example, it is now possible to produce transgenic
animals (e.g., mice) that are capable, upon immunization (or
reactive immunization in the case of catalytic antibodies) of
producing a full repertoire of human antibodies in the absence of
endogenous immunoglobulin production. For example, it has been
described that the homozygous deletion of the antibody heavy-chain
joining region (J.sub.H) gene in chimeric and germ-line
immunoglobulin gene array into such germ-line mutant mice will
result in the production of human antibodies upon antigen
challenge. See, e.g., B. D. Cohen et al, Clin. Cancer Res.
11:2063-2073 (2005); J. L. Teeling et al., Blood 104:1793-1800
(2004); N. Lonberg et al., Nature 368:856-859 (1994); A. Jakobovits
et al., Proc. Natl. Acad. Sci. U.S.A. 90:2551-2555 (1993); A.
Jakobovits et al., Nature 362:255-258 (1993); M. Bruggemann et al.,
Year Immunol. 7:33-40 (1993); L. D. Taylor, et al. Nucleic Acids
Res. 20:6287-6295 (1992); M. Bruggemann et al., Proc. Natl. Acad.
Sci. U.S.A. 86:6709-6713 (1989)); and WO 97/17852.
[0430] Alternatively, phage display technology (see, e.g., J.
McCafferty et al., Nature 348:552-553 (1990); H. J. de Haard et
al., J Biol Chem 274, 18218-18230 (1999); and A. Kanppik et al., J
Mol Biol, 296, 57-86 (2000)) can be used to produce human
antibodies and antibody fragments in vitro using immunoglobulin
variable (V) domain gene repertoires from unimmunized donors.
According to this technique, antibody V domain genes are cloned
in-frame into either a major or minor coat protein gene of a
filamentous bacteriophage, such as M13 or fd, and displayed as
functional antibody fragments on the surface of the phage particle.
Because the filamentous particle contains a single-stranded DNA
copy of the phage genome, selections based on the functional
properties of the antibody also result in selection of the gene
encoding the antibody exhibiting those properties. Thus, the phage
mimics some of the properties of the B-cell. Phage display can be
performed in a variety of formats, and is reviewed in, e.g., K. S.
Johnson and D. J. Chiswell, Curr. Opin. Struct. Biol. 3:564-571
(1993). Several sources of V-gene segments can be used for phage
display. T. Clackson et al., Nature, 352:624-628 (1991) isolated a
diverse array of anti-oxazolone antibodies from a small random
combinatorial library of V genes derived from the spleens of
immunized mice. A repertoire of V genes from unimmunized human
donors can be constructed and antibodies to a diverse array of
antigens (including self-antigens) can be isolated essentially
following the techniques described by J. D. Marks et al., J. Mol.
Biol. 222:581-597 (1991) or A. D. Griffiths et al., EMBO J.
12:725-734 (1993). See also U.S. Pat. Nos. 5,565,332 and 5,573,905;
and L. S. Jespers et al., Biotechnology 12:899-903 (1994).
[0431] As indicated above, human antibodies may also be generated
by in vitro activated B cells. See, e.g., U.S. Pat. Nos. 5,567,610
and 5,229,275; and C. A. K. Borrebaeck et al., Proc. Natl. Acad.
Sci. U.S.A. 85:3995-3999 (1988).
[0432] Amino acid sequence modification(s) of the antibodies
described herein are contemplated. For example, it may be desirable
to improve the binding affinity and/or other biological properties
of the antibody. Amino acid sequence variants of an antibody are
prepared by introducing appropriate nucleotide changes into the
antibody nucleic acid, or by peptide synthesis. Such modifications
include, for example, deletions from, insertions into, and/or
substitutions of residues within the amino acid sequences of the
antibody. Any combination of deletion, insertion, and substitution
is made to arrive at the final construct, provided that the final
construct possesses the desired characteristics. The amino acid
changes also may alter post-translational processes of the
antibody, such as changing the number or position of glycosylation
sites.
[0433] A useful method for identification of certain residues or
regions of an antibody that are preferred locations for mutagenesis
is called "alanine scanning mutagenesis," as described in B. C.
Cunningham and J. A. Wells, Science 244:1081-1085 (1989). Here, a
residue or group of target residues are identified (e.g., charged
residues such as Arg, Asp, His, Lys, and Glu) and replaced by a
neutral or negatively charged amino acid (most preferably Ala or
Polyalanine) to affect the interaction of the amino acids with the
Z group of the linker. Those amino acid locations demonstrating
functional sensitivity to the substitutions are then refined by
introducing further or other variants at, or for, the sites of
substitution. Thus, while the site for introducing an amino acid
sequence variation is predetermined, the nature of the mutation per
se need not be predetermined. For example, to analyze the
performance of a mutation at a given site, alanine scanning or
random mutagenesis is conducted at the target codon or region and
the expressed antibody variants are screened for the ability to
form a covalent bond with Z.
[0434] Amino acid sequence insertions include amino- and/or
carboxyl-terminal fusions ranging in length from one residue to
polypeptides containing a hundred or more residues, as well as
intrasequence insertions of single or multiple amino acid residues.
Examples of terminal insertions include an antibody with an
N-terminal methionyl residue or the antibody fused to a cytotoxic
polypeptide. Other insertional variants of an antibody molecule
include the fusion to the N- or C-terminus of an anti-antibody to
an enzyme or a polypeptide which increases the serum half-life of
the antibody.
[0435] Another type of variant is an amino acid substitution
variant. These variants have at least one amino acid residue in an
antibody molecule replaced by a different residue. The sites of
greatest interest for substitutional mutagenesis include the
hypervariable regions, but FR alterations are also contemplated.
Conservative substitutions are shown in Table 3 below under the
heading of "preferred substitutions." If such substitutions result
in a change in biological activity, then more substantial changes,
denominated "exemplary substitutions" as further described below in
reference to amino acid classes, may be introduced and the products
screened.
[0436] Substantial modifications in the biological properties of
the antibody are accomplished by selecting substitutions that
differ significantly in their effect on maintaining (a) the
structure of the polypeptide backbone in the area of the
substitution, for example, as a sheet or helical conformation, (b)
the charge or hydrophobicity of the molecule at the target site, or
(c) the bulk of the side chain. Naturally occurring residues are
divided into groups based on common side-chain properties: [0437]
(1) hydrophobic: Nle, Met, Ala, Val, Leu, Ile; [0438] (2) neutral
hydrophilic: Cys, Ser, Thr; [0439] (3) acidic: Asp, Glu; [0440] (4)
basic: Asn, Gln, His, Lys, Arg; [0441] (5) residues that influence
chain orientation: Gly, Pro; and [0442] (6) aromatic: Trp, Tyr,
Phe.
[0443] Non-conservative substitutions will entail exchanging a
member of one of these classes for a member of another class.
[0444] Any cysteine residue not involved in maintaining the proper
conformation of the antibody may be substituted, generally with
serine, to improve the oxidative stability of the molecule and
prevent aberrant crosslinking. Conversely, cysteine bond(s) may be
added to the antibody to improve its stability (particularly where
the antibody is an antibody fragment such as an Fv fragment).
[0445] One type of substitutional variant involves substituting one
or more hypervariable region residues of a parent antibody (e.g., a
humanized or human antibody). Generally, the resulting variant(s)
selected for further development will have improved biological
properties relative to the parent antibody from which they are
generated. A convenient way for generating such substitutional
variants involves affinity maturation using phage display. Briefly,
several hypervariable region sites (e.g., 6-7 sites) are mutated to
generate all possible amino substitutions at each site. The
antibody variants thus generated are displayed in a monovalent
fashion from filamentous phage particles as fusions to the gene III
product of M13 packaged within each particle. The phage-displayed
variants are then screened for their biological activity (e.g.,
binding affinity) as herein disclosed. In order to identify
candidate hypervariable region sites for modification, alanine
scanning mutagenesis can be performed to identify hypervariable
region residues contributing significantly to antigen binding.
Alternatively, or additionally, it may be beneficial to analyze a
structure of the antibody conjugate complex to identify contact
points between the antibody and the Z group. Such contact residues
and neighboring residues are candidates for substitution according
to the techniques elaborated herein. Once such variants are
generated, the panel of variants is subjected to screening as
described herein and antibodies with superior properties in one or
more relevant assays may be selected for further development.
[0446] Another type of amino acid variant of the antibody alters
the original glycosylation pattern of the antibody by deleting one
or more carbohydrate moieties found in the antibody and/or adding
one or more glycosylation sites that are not present in the
antibody.
[0447] Glycosylation of antibodies is typically either N-linked or
O-linked. N-linked refers to the attachment of the carbohydrate
moiety to the side chain of an asparagine residue. The tripeptide
sequences Asn-X''-Ser and Asn-X''-Thr, where X'' is any amino acid
except proline, are generally the recognition sequences for
enzymatic attachment of the carbohydrate moiety to the asparagine
side chain. Thus, the presence of either of these tripeptide
sequences in a polypeptide creates a potential glycosylation site.
O-linked glycosylation refers to the attachment of one of the
sugars N-acetylgalactosamine, galactose, or xylose to a
hydroxyamino acid, most commonly serine or threonine, although
5-hydroxyproline or 5-hydroxylysine may also be used.
[0448] Addition of glycosylation sites to the antibody is
conveniently accomplished by altering the amino acid sequence such
that it contains one or more of the above-described tripeptide
sequences (for N-linked glycosylation sites). The alteration may
also be made by the addition of or substitution by one or more
serine or threonine residues to the sequence of the original
antibody (for O-linked glycosylation sites).
[0449] It may be desirable to modify an antibody with respect to
effector function, for example to enhance antigen-dependent
cell-mediated cytotoxicity (ADCC) and/or complement dependent
cytotoxicity (CDC) of the antibody. This may be achieved by
introducing one or more amino acid substitutions in an Fc region of
the antibody. Alternatively, an antibody can be engineered which
has dual Fc regions and may thereby have enhanced complement lysis
and ADCC capabilities. See G. T. Stevenson et al., Anticancer Drug
Des. 3:219-230 (1989).
[0450] To increase the serum half life of an antibody, one may
incorporate a salvage receptor binding epitope into the antibody
(especially an antibody fragment) as described in U.S. Pat. No.
5,739,277, for example. As used herein, the term "salvage receptor
binding epitope" refers to an epitope of the Fc region of an IgG
molecule (e.g., IgG.sub.1, IgG.sub.2, IgG.sub.3, or IgG.sub.4) that
is responsible for increasing the in vivo serum half-life of the
IgG molecule.
TABLE-US-00022 TABLE 3 Amino acid substitutions Original Residue
Exemplary Substitutions Preferred Substitutions Ala (A) Val; Leu;
Ile Val Arg I Lys; Gln; Asn Lys Asn (N) Gln; His; Asp; Lys; Arg Gln
Asp (D) Glu; Asn Glu Cl(C) Ser; Ala Ser Gln (Q) Asn; Glu Asn Glu
(E) Asp; Gln Asp Gly (G') Ala Ala His (H) Asn; Gln; Lys; Arg Arg
Ile (I) Leu; Val; Met; Ala; Phe; Nle Leu Leu (L) Nle; Ile; Val;
Met; Ala; Phe Ile Lys (K) Arg; Gln; Asn Arg Met (M) Leu; Phe; Ile
Leu Phe (F) Leu; Val; Ile; Ala; Tyr Tyr Pro (P) Ala Ala Ser (S) Thr
Thr Thr (T) Ser Ser Trp (W) Tyr; Phe Tyr Tyr (Y) Trp; Phe; Thr; Ser
Phe Val (V) Ile; Leu; Met; Phe; Ala; Nle Leu
[0451] Various techniques have been developed for the production of
whole antibodies and antibody fragments. Traditionally, antibody
fragments were derived via proteolytic digestion of intact
antibodies (see, e.g., K. Morimoto and K. Inouye, J. Biochem.
Biophys. Methods 24:107-117 (1992); M. Brennan et al., Science
229:81-83 (1985)). However, these fragments can now be produced
directly by recombinant host cells. Fab, Fv, V.sub.H, V.sub.L, and
scFv antibody fragments can all be expressed in and secreted from
E. coli as is detailed below, thus allowing the facile production
of large amounts of these fragments. Antibody fragments can be
isolated from the antibody phage libraries discussed above.
Alternatively, Fab'-SH fragments can be directly recovered from E.
coli and chemically coupled to form F(ab').sub.2 fragments (P.
Carter et al., Biotechnology 10:163-167 (1992)). According to
another approach, F(ab').sub.2 fragments can be isolated directly
from recombinant host cell culture.
[0452] A variety of expression vector/host systems may be utilized
to express antibodies. These systems include but are not limited to
microorganisms such as bacteria transformed with recombinant
bacteriophage, plasmid or cosmid DNA expression vectors; yeast
transformed with yeast expression vectors; insect cell systems
infected with virus expression vectors (e.g., baculovirus); plant
cell systems transfected with virus expression vectors (e.g.,
cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or
transformed with bacterial expression vectors (e.g., Ti or pBR322
plasmid); or animal cell systems.
[0453] Mammalian cells that are useful in recombinant antibody
expression include but are not limited to VERO cells, HeLa cells,
Chinese hamster ovary (CHO) cell lines, COS cells (such as COS-7),
W138, BHK, HepG2, 3T3, RIN, MDCK, A549, PC12, K562 and 293 cells,
as well as hybridoma cell lines as described herein. Mammalian
cells are preferred for preparation of those antibodies that are
typically glycosylated and require proper refolding for activity.
Preferred mammalian cells include CHO cells, hybridoma cells, and
myeloid cells.
[0454] Some exemplary protocols for the recombinant expression of
antibodies are described herein below.
[0455] The term "expression vector" or "vector" refers to a
plasmid, phage, virus or vector, for expressing a polypeptide from
a DNA (RNA) sequence. An expression vector may comprise a
transcriptional unit comprising (1) one or more regulatory
sequences controlling gene expression, for example, promoters or
enhancers, (2) one or more sequences that encode one or more
polypeptides, and (3) appropriate transcription initiation and
termination sequences. Expression vectors intended for use in yeast
or eukaryotic expression systems preferably include a leader
sequence enabling extracellular secretion of translated protein by
a host cell. Alternatively, where an antibody polypeptide(s) is
expressed without a leader or transport sequence, it may include an
amino terminal methionine residue. This residue may or may not be
subsequently cleaved from the expressed recombinant protein to
provide a final antibody product.
[0456] Antibodies, specifically antibody fragments, may be
expressed in prokaryotic systems such as E. coli. In another
example, the DNA sequence encoding the specific binding agent
peptide can be amplified by PCR and cloned into an appropriate
vector, such as for example pGEX-3x (Pharmacia). The pGEX vector is
designed to produce a fusion protein comprising
glutathione-S-transferase (GST), encoded by the vector, and a
peptide encoded by a DNA fragment inserted into the vector's
cloning site. The primers for PCR can be generated to include for
example, an appropriate cleavage site. The pGEX-3x antibody peptide
construct is transformed into E. coli XL-1 Blue cells (Stratagene,
La Jolla Calif.), and individual transformants are isolated and
grown. The expressed peptide fusion protein may then be cleaved
from the GST portion of the fusion protein.
[0457] Expression of polynucleotides encoding antibodies using the
recombinant systems described above may result in production of
antibodies or fragments thereof that must be "re-folded" (to
properly create various disulphide bridges) in order to be
biologically active.
[0458] Antibodies, specifically antibody fragments, made in
bacterial cells may be produced as an insoluble inclusion body in
the bacteria. Such antibodies can be purified as follows. Host
cells can be sacrificed by centrifugation; washed in 0.15 M NaCl,
10 mM Tris, pH 8, 1 mM EDTA; and treated with 0.1 mg/ml lysozyme
(Sigma, St. Louis, Mo.) for 15 minutes at room temperature. The
lysate can be cleared by sonication, and cell debris can be
pelleted by centrifugation for 10 minutes at 12,000.times.g. The
antibody containing pellet can be resuspended in 50 mM Tris, pH 8,
and 10 mM EDTA, layered over 50% glycerol, and centrifuged for 30
min. at 6000.times.g. The pellet can be resuspended in standard
phosphate buffered saline solution (PBS) free of Mg and Ca ions.
The antibody can be further purified by fractionating the
resuspended pellet in a denaturing SDS polyacrylamide gel (Sambrook
et al., supra). The gel can be soaked in 0.4 M KCl to visualize the
protein, which can be excised and electroeluted in gel-running
buffer lacking SDS.
[0459] Mammalian host systems for the expression of antibodies are
well known to those of skill in the art. Host cell strains can be
chosen for a particular ability to process the expressed protein or
produce certain post-translation modifications that will be useful
in providing protein activity. Such modifications of the
polypeptide include, but are not limited to, acetylation,
carboxylation, glycosylation, phosphorylation, lipidation and
acylation. Different host cells such as CHO, HeLa, MDCK, 293, W138,
as well as hybridoma cell lines, and the like have specific
cellular machinery and characteristic mechanisms for such
post-translational activities and can be chosen to ensure the
correct modification and processing of the introduced, foreign
protein.
[0460] A number of selection systems can be used to recover the
cells that have been transformed for recombinant antibody
production. Such selection systems include, but are not limited to,
HSV thymidine kinase, hypoxanthine-guanine
phosphoribosyltransferase and adenine phosphoribosyltransferase
genes, in tk-, hgprt- or aprt-cells, respectively. Also,
anti-metabolite resistance can be used as the basis of selection
for DHFR which confers resistance to methotrexate; gpt which
confers resistance to mycophenolic acid; neo which confers
resistance to the aminoglycoside G418 and confers resistance to
chlorsulfuron; and hygro which that confers resistance to
hygromycin. Additional selectable genes that may be useful include
trpB, which allows cells to utilize indole in place of tryptophan,
or hisD, which allows cells to utilize histinol in place of
histidine. Markers that give a visual indication for identification
of transformants include anthocyanins, beta.-glucuronidase and its
substrate, GUS, and luciferase and its substrate, luciferin.
[0461] In some cases, antibodies produced using procedures
described above may need to be "refolded" and oxidized into a
proper tertiary structure and allowed to generate disulfide
linkages in order to be biologically active. Refolding can be
accomplished using a number of procedures well known in the art.
Such methods include, for example, exposing the solubilized
polypeptide agent to a pH usually above 7 in the presence of a
chaotropic agent. The selection of chaotrope is similar to the
choices used for inclusion body solubilization. However a chaotrope
is typically used at a lower concentration. An exemplary chaotropic
agent is guanidine. In most cases, the refolding/oxidation solution
will also contain a reducing agent plus its oxidized form in a
specific ratio to generate a particular redox potential which
allows for disulfide shuffling to occur for the formation of
cysteine bridges. Some commonly used redox couples include
cysteine/cystamine, glutathione/dithiobisGSH, cupric chloride,
dithiothreitol DTT/dithiane DTT, and 2-mercaptoethanol
(bME)/dithio-bME. In many instances, a co-solvent may be used to
increase the efficiency of the refolding. Commonly used cosolvents
include glycerol, polyethylene glycol of various molecular weights,
and arginine.
Linkers and Linked Compounds
[0462] An AA targeting agent may be covalently linked to a
combining site in an antibody either directly or via a linker. An
appropriate linker can be chosen to provide sufficient distance
between the targeting agent and the antibody The general design of
an embodiment of a linker for use in preparing AA targeting
compounds is represented by the formula: --X-Recognition GroupY--Z,
wherein X is a connecting chain, Recognition Group Y is a
recognition group and Z is a reactive group. The linker may be
linear or branched, and optionally includes one or more carbocyclic
or heterocyclic groups. Linker length may be viewed in terms of the
number of linear atoms, with cyclic moieties such as aromatic rings
and the like to be counted by taking the shortest route around the
ring. In some embodiments, the linker has a linear stretch of
between 5-15 atoms, in other embodiments 15-30 atoms, in still
other embodiments 30-50 atoms, in still other embodiments 50-100
atoms, and in still other embodiments 100-200 atoms. Other linker
considerations include the effect on physical or pharmacokinetic
properties of the resulting AA targeting compound or AA targeting
agent-linker, such as solubility, lipophilicity, hydrophilicity,
hydrophobicity, stability (more or less stable as well as planned
degradation), rigidity, flexibility, immunogenicity, modulation of
antibody binding, the ability to be incorporated into a micelle or
liposome, and the like.
[0463] The connecting chain X of the linker includes any atom from
the group C, H, N, O, P, S, halogen (F, Cl, Br, I), or a salt
thereof. X also may include a group such as an alkyl, alkenyl,
alkynyl, oxoalkyl, oxoalkenyl, oxoalkynyl, aminoalkyl,
aminoalkenyl, aminoalkynyl, sulfoalkyl, sulfoalkenyl, sulfoalkynyl,
phosphoalkyl, phosphoalkenyl, or phosphoalkynyl group. In some
embodiments, X may include one or more ring structures. In some
embodiments, the linker is a repeating polymer such as polyethylene
glycol comprising 2-100 units.
[0464] The recognition group Y of the linker is optional, and if
present is located between the reactive group and the connecting
chain. In some embodiments, Recognition Group Y is located from
1-20 atoms from Z. Although not wishing to be bound by any theory,
it is believed that the recognition group acts to properly position
the reactive group into the antibody combining site so that it may
react with a reactive amino acid side chain. Exemplary recognition
groups include carbocyclic and heterocyclic rings, preferably
having five or six atoms. However, larger ring structures also may
be used. In some embodiments, an AA targeting agent is linked
directly to Recognition Group Y without the use of an intervening
linker.
[0465] Z is capable of forming a covalent bond with a reactive side
chain in an antibody combining site. In some embodiments, Z
includes one or more C.dbd.O groups arranged to form a diketone, an
acyl beta-lactam, an active ester, a haloketone, a cyclohexyl
diketone group, an aldehyde, a maleimide, an activated alkene, an
activated alkyne or, in general, a molecule comprising a leaving
group susceptible to nucleophilic or electrophilic displacement.
Other groups may include a lactone, an anhydride, an
alpha-haloacetamide, an imine, a hydrazide, or an epoxide.
Exemplary linker electrophilic reactive groups that can covalently
bond to a reactive nucleophilic group (e.g., a lysine or cysteine
side chain) in a combining site of antibody include acyl
beta-lactam, simple diketone, succinimide active ester, maleimide,
haloacetamide with linker, haloketone, cyclohexyl diketone,
aldehyde, amidine, guanidine, imine, eneamine, phosphate,
phosphonate, epoxide, aziridine, thioepoxide, a masked or protected
diketone (a ketal for example), lactam, sulfonate, and the like,
masked C.dbd.O groups such as imines, ketals, acetals, and any
other known electrophilic group. In certain embodiments, the
reactive group includes one or more C.dbd.O groups arranged to form
an acyl beta-lactam, simple diketone, succinimide active ester,
maleimide, haloacetamide with linker, haloketone, cyclohexyl
diketone, or aldehyde.
[0466] The linker reactive group or similar such reactive group is
chosen for use with a reactive residue in a particular combining
site. For example, a chemical moiety for modification by an
aldolase antibody may be a ketone, diketone, beta lactam, active
ester haloketone, lactone, anhydride, maleimide,
alpha-haloacetamide, cyclohexyl diketone, epoxide, aldehyde,
amidine, guanidine, imine, eneamine, phosphate, phosphonate,
epoxide, aziridine, thioepoxide, masked or protected diketone
(ketal for example), lactam, haloketone, aldehyde, and the
like.
[0467] A linker reactive group chemical moiety suitable for
covalent modification by a reactive sulfhydryl group in an antibody
may be a disulfide, aryl halide, maleimide, alpha-haloacetamide,
isocyanate, epoxide, thioester, active ester, amidine, guanidine,
imine, eneamine, phosphate, phosphonate, epoxide, aziridine,
thioepoxide, masked or protected diketone (ketal for example),
lactam, haloketone, aldehyde, and the like.
[0468] One of skill in the art will readily appreciate that
reactive amino acid side chains in antibody combining sites may
possess an electrophilic group that reacts with a nucleophilic
group on an AA targeting agent or its linker, whereas in other
embodiments a reactive nucleophilic group in an amino acid side
chain reacts with an electrophilic group in an AA targeting agent
or linker.
[0469] An AA targeting compound may be prepared by several
approaches. In one approach, an AA targeting agent-linker compound
is synthesized with a linker that includes one or more reactive
groups designed for covalent reaction with a side chain of an amino
acid in a combining site of an antibody. The targeting agent-linker
compound and antibody are combined under conditions where the
linker reactive group forms a covalent bond with the amino acid
side chain.
[0470] In another approach, linking can be achieved by synthesizing
an antibody-linker compound comprising an antibody and a linker
wherein the linker includes one or more reactive groups designed
for covalent reaction with an appropriate chemical moiety of an AA
targeting agent. An AA targeting agent may need to be modified to
provide the appropriate moiety for reaction with the linker
reactive group. The antibody-linker and AA targeting agent are
combined under conditions where the linker reactive group
covalently links to the targeting and/or biological agent.
[0471] A further approach for forming an antibody-AA targeting
compound uses a dual linker design. In certain embodiments, an AA
targeting agent-linker compound is synthesized which comprises an
AA targeting agent and a linker with a reactive group. An
antibody-linker compound is synthesized which comprises an antibody
and a linker with a chemical group susceptible to reactivity with
the reactive group of the AA targeting agent-linker of the first
step. These two linker containing compounds are then combined under
conditions whereby the linkers covalently link, forming the
antibody-AA-targeting compound.
[0472] Exemplary functional groups that can be involved in the
linkage include, for example, esters, amides, ethers, phosphates,
amino, keto, amidine, guanidine, imines, eneamines, phosphates,
phosphonates, epoxides, aziridines, thioepoxides, masked or
protected diketones (ketals for example), lactams, haloketones,
aldehydes, thiocarbamate, thioamide, thioester, sulfide, disulfide,
phosphoramide, sulfonamide, urea, thioruea, carbamate, carbonate,
hydroxamide, and the like.
[0473] The linker includes any atom from the group C, H, N, O, P,
S, halogen (F, Cl, Br, I), or a salt thereof. The linker also may
include a group such as an alkyl, alkenyl, alkynyl, oxoalkyl,
oxoalkenyl, oxoalkynyl, aminoalkyl, aminoalkenyl, aminoalkynyl,
sulfoalkyl, sulfoalkenyl, sulfoalkynyl group, phosphoalkyl,
phosphoalkenyl, or phosphoalkynyl group. The linker also may
include one or more ring structures. As used herein, a "ring
structure" includes saturated, unsaturated, and aromatic
carbocyclic rings and saturated, unsaturated, and aromatic
heterocyclic rings. The ring structures may be mono-, bi-, or
polycyclic, and include fused or unfused rings. Further, the ring
structures are optionally substituted with functional groups well
known in the art, including but not limited to halogen, oxo, --OH,
--CHO, --COOH, --NO.sub.2, --CN, --NH.sub.2, --C(O)NH.sub.2,
C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.1-6
oxoalkyl, oxoalkenyl, oxoalkynyl, aminoalkyl, aminoalkenyl,
aminoalkynyl, sulfoalkyl, sulfoalkenyl, sulfoalkynyl, phosphoalkyl,
phosphoalkenyl, or phosphoalkynyl group. Combinations of the above
groups and rings may also be present in the linkers of AA targeting
compounds.
[0474] One aspect of the invention is an AA targeting agent-linker
conjugate having Formula I:
Linker-[AA targeting agent] (I)
wherein [AA targeting agent] is an AA targeting agent peptide.
[0475] The linker moiety Linker in compounds of Formula I may be
attached to the amino terminus, carboxy terminus or any amino acid
side chain of an AA targeting agent. In certain embodiments, Linker
is linked to the carboxy terminus of an AA targeting agent. In
certain other embodiments, Linker is linked to the amino terminus
of an AA targeting agent. In still other embodiments, Linker is
linked to either a nucleophilic or electrophilic side chain. For
the case of linking to an electrophilic side chain, Linker should
possess a nucleophilic group susceptible to covalent reaction with
the electrophilic side chain. Exemplary electrophilic side chains
are Asp and Glu. Exemplary nucleophilic side chains are Cys, Lys,
Ser, Thr, and Tyr. For the case of linking to a nucleophilic side
chain, Linker should comprise an electrophilic group susceptible to
covalent reaction with the nucleophilic side chain. In another
embodiment, a nucleophilic amino acid is added to either the
carboxy terminus or the amino terminus of an AA targeting agent and
the linker Linker is covalently attached to the side chain of this
additional amino acid. In certain embodiments, Lys is added to the
amino terminus of an AA targeting agent. In certain other
embodiments, Lys is added to the carboxy terminus of an AA
targeting agent.
[0476] Thus, in those embodiments comprising R.sup.1-QKY QPL DEL
DKT LYD QFM LQQ G-R.sup.2 (SEQ ID NO: 21) based AA targeting
agents, exemplary compounds of Formula I formed by linking to
either i) the side chains of D, E, K, T, and Y or ii) the amino or
carboxy termini, include the following, where (Li) following an
amino acid residue indicates linking to that residue: [0477]
Q(Li)KY QPL DEL DKT LYD QFM LQQ G-R.sup.2 (SEQ ID NO:140) [0478]
R.sup.1-QK(Li)Y QPL DEL DKT LYD QFM LQQ G-R.sup.2 (SEQ ID NO:141)
[0479] R.sup.1-QKY(Li) QPL DEL DKT LYD QFM LQQ G-R.sup.2 (SEQ ID
NO:142) [0480] R.sup.1-QKY QPL D(Li)EL DKT LYD QFM LQQ G-R.sup.2
(SEQ ID NO:13) [0481] R.sup.1-QKY QPL DE(Li)L DKT LYD QFM LQQ
G-R.sup.2 (SEQ ID NO:144) [0482] R.sup.1-QKY QPL DEL D(Li)KT LYD
QFM LQQ G-R.sup.2 (SEQ ID NO:145) [0483] R.sup.1-QKY QPL DEL
DK(Li)T LYD QFM LQQ G-R.sup.2 (SEQ ID NO:146) [0484] R.sup.1-QKY
QPL DEL DKT(Li) LYD QFM LQQ G-R.sup.2 (SEQ ID NO:147) [0485]
R.sup.1-QKY QPL DEL DKT LY(Li)D QFM LQQ G-R.sup.2 (SEQ ID NO:148)
[0486] R.sup.1-QKY QPL DEL DKT LYD(Li) QFM LQQ G-R.sup.2 (SEQ ID
NO:149) [0487] R.sup.1-QKY QPL DEL DKT LYD QFM LQQ G(Li) (SEQ ID
NO:150)
[0488] Similarly, in those embodiments comprising R.sup.1-Q-[ACK]-Y
QPL DEL DKT LYD QFM LQQ G-R.sup.2 (SEQ ID NO: 29) based AA
targeting agents, exemplary compounds of Formula I formed by
linking to either i) the side chains of D, E, K, T, and Y or ii)
the amino or carboxy termini, include: [0489] Q(Li)-[AcK]Y QPL DEL
DKT LYD QFM LQQ G-R.sup.2 (SEQ ID NO:151) [0490]
R.sup.1-Q-[AcK]-Y(Li) QPL DEL DKT LYD QFM LQQ G-R.sup.2 (SEQ ID
NO:152) [0491] R.sup.1-Q-[AcK]-Y QPL D(Li)EL DKT LYD QFM LQQ
G-R.sup.2 (SEQ ID NO:153) [0492] R.sup.1-Q-[AcK]-Y QPL DE(Li)L DKT
LYD QFM LQQ G-R.sup.2 (SEQ ID NO:154) [0493] R.sup.1-Q-[AcK]-Y QPL
DEL D(Li)KT LYD QFM LQQ G-R.sup.2 (SEQ ID NO:155) [0494]
R.sup.1-Q-[AcK]-Y QPL DEL DK(Li)T LYD QFM LQQ G-R.sup.2 (SEQ ID
NO:156) [0495] R.sup.1-Q-[AcK]-Y QPL DEL DKT(Li) LYD QFM LQQ
G-R.sup.2 (SEQ ID NO:157) [0496] R.sup.1-Q-[AcK]-Y QPL DEL DKT
LY(Li)D QFM LQQ G-R.sup.2 (SEQ ID NO:158) [0497] R.sup.1-Q-[AcK]-Y
QPL DEL DKT LYD(Li) QFM LQQ G-R.sup.2 (SEQ ID NO:159) [0498]
R.sup.1-Q-[AcK]-Y QPL DEL DKT LYD QFM LQQ G(Li) (SEQ ID NO:160)
[0499] In compounds of Formula I, Linker is a linker moiety having
the formula --X-Recognition Group Y--Z, wherein: [0500] X is an
optionally present biologically compatible polymer or block
copolymer attached to one of the residues that comprises an AA
targeting agent; [0501] Y is an optionally present recognition
group comprising at least a ring structure; and [0502] Z is a
reactive group that is capable of covalently linking to a side
chain in a combining site of an antibody.
[0503] In some embodiments of compounds in Formula I, X is:
--R.sup.22--P--R.sup.2-- or
--R.sup.22--P--R.sup.21--P--R.sup.23--
wherein: [0504] P and P' are independently selected from the group
consisting of polyoxyalkylene oxides such as polyethylene oxide,
polyethyloxazoline, poly-N-vinyl pyrrolidone, polyvinyl alcohol,
polyhydroxyethyl acrylate, polyhydroxy ethylmethacrylate and
polyacrylamide, polyamines having amine groups on either the
polymer backbone or the polymer sidechains, such as polylysine,
polyornithine, polyarginine, and polyhistidine, nonpeptide
polyamines such as polyaminostyrene, polyaminoacrylate,
poly(N-methyl aminoacrylate), poly(N-ethylaminoacrylate),
poly(N,N-dimethyl aminoacrylate), poly(N,N-diethylaminoacrylate),
poly(aminomethacrylate), poly(N-methyl aminomethacrylate),
poly(N-ethyl aminomethacrylate), poly(N,N-dimethyl
aminomethacrylate), poly(N,N-diethyl aminomethacrylate),
poly(ethyleneimine), polymers of quaternary amines, such as
poly(N,N,N-trimethylaminoacrylate chloride),
poly(methyacrylamidopropyltrimethyl ammonium chloride),
proteoglycans such as chondroitin sulfate-A (4-sulfate) chondroitin
sulfate-C(6-sulfate) and chondroitin sulfate-B, polypeptides such
as polyserine, polythreonine, polyglutamine, natural or synthetic
polysaccharides such as chitosan, hydroxy ethyl cellulose, and
lipids; [0505] R.sup.21, R.sup.22, and R.sup.23 are each
independently a covalent bond, --O--, --S--, --NR.sup.b--,
substituted or unsubstituted straight or branched chain C.sub.1-50
alkylene, or substituted or unsubstituted straight or branched
chain C.sub.1-50 heteroalkylene; [0506] R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.m alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl; and [0507] R.sup.21, R.sup.22,
and R.sup.23 are selected such that the backbone length of X
remains about 200 atoms or less.
[0508] In some embodiments of compounds of Formula I, R.sup.22 is
--(CH.sub.2).sub.v--, --(CH.sub.2).sub.u--C(O)--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(O)--O--(CH-.sub.2).sub.v,
--(CH.sub.2).sub.u--C(S)--NR.sup.b--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(O)--NR.sup.b--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--NR.sup.b--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--O--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--S(O).sub.0-2--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--S(O).sub.0-2--NR.sup.b--(CH.sub.2).sub.v--, or
--(CH.sub.2).sub.u--P(O)(OR.sup.b)--O--(CH.sub.2).sub.v--, wherein
u and v are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,
11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.
[0509] In yet other embodiments of compounds of Formula I, R.sup.22
is --(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(O)--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(O)--O--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(O)--NR.sup.b--(CH.sub.2).sub.v--, or
--(CH.sub.2).sub.u--NR.sup.b--(CH.sub.2).sub.v. In still other
embodiments, R.sup.-2 is
--(CH.sub.2).sub.u--C(O)--NR.sup.b--(CH.sub.2).sub.v--.
[0510] In some embodiments of compounds of Formula I, R.sup.21 and
R.sup.23 are each independently --(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--C(O)--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--C(O)--O--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.r--C(S)--NR.sup.b--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--C(O)--NR.sup.b--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--NR.sup.b--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--O--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--S(O).sub.0-2--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--S(O).sub.0-2--NR.sup.b--(CH.sub.2).sub.s--, or
--(CH.sub.2).sub.r--P(O)(OR.sup.b)--O--(CH.sub.2).sub.s--, wherein
r, s, and v are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9,
10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.
[0511] In yet other embodiments, R.sup.21 and R.sup.23 are each
independently --(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--C(O)--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--C(O)--O--(CH.sub.2).sub.s--,
--(CH.sub.2).sub.r--C(O)--NR.sup.b--(CH.sub.2).sub.s--, or
--(CH.sub.2).sub.r--NR.sup.b--(CH.sub.2).sub.s, and
--(CH.sub.2).sub.r--C(O)--NR.sup.b--(CH.sub.2).sub.s--.
[0512] In still other embodiments, R.sup.21 and R.sup.23 each
independently have the structure:
##STR00032##
wherein p is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,
17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,
34, 35, 36, 37, 38, 39, 40, 41, 32, 43, 44, or 45; w, r, and s are
each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,
14, 15, 16, 17, 18, 19 or 20; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl.
[0513] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00033##
wherein H.sup.1 and H.sup.1' at each occurrence are independently
N, O, S, or CH.sub.2; r and s are each independently 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; t and t'
are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,
30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 32, 43, 44, 45, 46,
47, 48, 49 or 50; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl.
[0514] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00034##
wherein H.sup.1 and H.sup.1' at each occurrence are independently
N, O, S, or CH.sub.2; r and s are each independently 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; t and t'
are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,
30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 32, 43, 44, 45, 46,
47, 48, 49 or 50; and R.sup.b at each occurrence is independently
hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl.
[0515] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00035##
wherein H.sup.1 and H.sup.1' at each occurrence are independently
N, O, S, or CH.sub.2; r and s are each independently 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; t and t'
are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,
30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 32, 43, 44, 45, 46,
47, 48, 49 or 50; and R.sup.b at each occurrence is independently
hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl.
[0516] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00036##
wherein H.sup.1 and H.sup.1' at each occurrence are independently
N, O, S, or CH.sub.2; r and s are each independently 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; t and t'
are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,
30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 32, 43, 44, 45, 46,
47, 48, 49 or 50; and R.sup.b at each occurrence is independently
hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl.
[0517] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00037##
wherein H.sup.1 and H.sup.1' at each occurrence are independently
N, O, S, or CH.sub.2; r and s are each independently 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; t and t'
are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,
30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 32, 43, 44, 45, 46,
47, 48, 49 or 50; and R.sup.b at each occurrence is independently
hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl.
[0518] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00038##
wherein H.sup.1 and H.sup.1' at each occurrence are independently
N, O, S, or CH.sub.2; r and s are each independently 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; t and t'
are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,
30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 32, 43, 44, 45, 46,
47, 48, 49 or 50; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl.
[0519] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00039##
wherein H.sup.1 and H.sup.1' at each occurrence are independently
N, O, S, or CH.sub.2; r and s are each independently 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; t and t'
are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,
30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 32, 43, 44, 45, 46,
47, 48, 49 or 50; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl.
[0520] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00040##
wherein v and w are each independently 1, 2, 3, 4, or 5 and R.sup.b
is hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain of
these embodiments, v is 1, 2 or 3, w is 1, 2, or 3, and R.sup.b is
hydrogen.
[0521] In certain embodiments of Formula I, Linker is a linker
moiety having the formula --X-Recognition GroupY--Z, wherein:
[0522] X is attached to one of the residues that comprises an AA
targeting agent, and is an optionally substituted
--R.sup.22--[CH.sub.2--CH.sub.2--O].sub.t--R.sup.23--,
--R.sup.22-cycloalkyl-R.sup.23--, --R.sup.22-aryl-R.sup.23--, or
--R.sup.22-heterocyclyl-R.sup.23--, wherein; [0523] R.sup.22 and
R.sup.23 are each independently a covalent bond, --O--, --S--,
--NR.sup.b--, substituted or unsubstituted straight or branched
chain C.sub.1-50 alkylene, substituted or unsubstituted straight or
branched chain C.sub.1-50 heteroalkylene, substituted or
unsubstituted straight or branched chain C.sub.2-50 alkenylene, or
substituted or unsubstituted C.sub.2-50 heteroalkenylene; [0524]
R.sup.b is hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl; [0525] t is
0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,
19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,
36, 37, 38, 39, 40, 41, 32, 43, 44, 45, 46, 47, 48, 49 or 50;
[0526] and the size of R.sup.22 and R.sup.23 are such that the
backbone length of X remains about 200 atoms or less; [0527]
Recognition Group Y is an optionally present recognition group
comprising at least a ring structure; and [0528] Z is a reactive
group that is capable of covalently linking to a side chain in a
combining site of an antibody. In some embodiments of compounds of
Formula I, if t>1 or if --X is --R.sup.22-cycloalkyl-R.sup.23--,
--R.sup.22-aryl-R.sup.23--, or --R.sup.22-heterocyclyl-R.sup.23--,
Recognition Group Y is present.
[0529] In some embodiments of compounds of Formula I, X is:
--R.sup.22--[CH.sub.2--CH.sub.2--O].sub.t--R.sup.23--,
wherein: [0530] R.sup.22 is --(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(O)--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(O)--O--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(O)--NR.sup.b--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--C(S)--NR.sup.b--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--NR.sup.b--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--O--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--S(O).sub.0-2--(CH.sub.2).sub.v--,
--(CH.sub.2).sub.u--S(O).sub.0-2--NR.sup.b--(CH.sub.2).sub.v--, or
--(CH.sub.2).sub.u--P(O)(OR.sup.b)--O--(CH.sub.2).sub.v--; [0531] u
and v are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,
12, 13, 14, 15, 16, 17, 18, 19 or 20 and t is 0 to 50. [0532]
R.sup.23 has the structure:
##STR00041##
[0532] wherein: [0533] p is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,
30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 32, 43, 44, or 45;
[0534] w and r are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9,
10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20; [0535] s is 0, 1, 2,
3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;
and [0536] R.sup.b at each occurrence is independently hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl; [0537] and the values of t,
u, w, p, v, r and s are such that the backbone length of X remains
about 200 atoms or less.
[0538] In certain embodiments of compounds of Formula I, X has the
formula:
##STR00042##
wherein the values of v, t, w, and p are selected such that the
backbone length of X is less than 200 atoms, alternatively is less
than 100 atoms, alternatively is less than 75 atoms, alternatively
is less than 50 atoms, alternatively is less than 25 atoms, or
alternatively is less than 15 atoms.
[0539] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00043##
wherein the values of v, t, r, and s are selected such that the
backbone length of X is less than 200 atoms, alternatively is less
than 100 atoms, alternatively is less than 75 atoms, alternatively
is less than 50 atoms, alternatively is less than 25 atoms, or
alternatively is less than 15 atoms.
[0540] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00044##
wherein the values of u, v, t, w, and p are selected such that the
backbone length of X is less than 200 atoms, alternatively is less
than 100 atoms, alternatively is less than 75 atoms, alternatively
is less than 50 atoms, alternatively is less than 25 atoms, or
alternatively is less than 15 atoms.
[0541] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00045##
wherein the values of u, v, t, r, and s are selected such that the
backbone length of X is less than 200 atoms, alternatively is less
than 100 atoms, alternatively is less than 75 atoms, alternatively
is less than 50 atoms, alternatively is less than 25 atoms, or
alternatively is less than 15 atoms.
[0542] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00046##
wherein the values of u, v, t, w, and p are selected such that the
backbone length of X is less than 200 atoms, alternatively is less
than 100 atoms, alternatively is less than 75 atoms, alternatively
is less than 50 atoms, alternatively is less than 25 atoms, or
alternatively is less than 15 atoms.
[0543] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00047##
wherein the values of u, v, t, r, and s are selected such that the
backbone length of X is less than 200 atoms, alternatively is less
than 100 atoms, alternatively is less than 75 atoms, alternatively
is less than 50 atoms, alternatively is less than 25 atoms, or
alternatively is less than 15 atoms.
[0544] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00048##
wherein the values of u, v, t, w, and p are selected such that the
backbone length of X is less than 200 atoms, alternatively is less
than 100 atoms, alternatively is less than 75 atoms, alternatively
is less than 50 atoms, alternatively is less than 25 atoms, or
alternatively is less than 15 atoms.
[0545] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00049##
wherein the values of u, v, t, r, and s are selected such that the
backbone length of X is less than 200 atoms, alternatively is less
than 100 atoms, alternatively is less than 75 atoms, alternatively
is less than 50 atoms, alternatively is less than 25 atoms, or
alternatively is less than 15 atoms.
[0546] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00050##
wherein the values of u, v, t, w, and p are selected such that the
backbone length of X is less than 200 atoms, alternatively is less
than 100 atoms, alternatively is less than 75 atoms, alternatively
is less than 50 atoms, alternatively is less than 25 atoms, or
alternatively is less than 15 atoms.
[0547] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00051##
wherein the values of u, v, t, r, and s are selected such that the
backbone length of X is less than 200 atoms, alternatively is less
than 100 atoms, alternatively is less than 75 atoms, alternatively
is less than 50 atoms, alternatively is less than 25 atoms, or
alternatively is less than 15 atoms.
[0548] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00052##
wherein the values of u, v, t, w, and p are selected such that the
backbone length of X is less than 200 atoms, alternatively is less
than 100 atoms, alternatively is less than 75 atoms, alternatively
is less than 50 atoms, alternatively is less than 25 atoms, or
alternatively is less than 15 atoms.
[0549] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00053##
wherein the values of u, v, t, r, and s are selected such that the
backbone length of X is less than 200 atoms, alternatively is less
than 100 atoms, alternatively is less than 75 atoms, alternatively
is less than 50 atoms, alternatively is less than 25 atoms, or
alternatively is less than 15 atoms.
[0550] In compounds having Formula I wherein Linker has the formula
--X-Recognition GroupY--Z, the ring structure of Recognition Group
Y includes saturated, unsaturated, and aromatic carbocyclic rings
and saturated, unsaturated, and aromatic heterocyclic rings. The
ring structure(s) may be mono-, bi-, or polycyclic, and include
fused or unfused rings. Further, the ring structure(s) is
optionally substituted with functional groups well known in the art
including, but not limited to halogen, oxo, --OH, --CHO, --COOH,
--NO.sub.2, --CN, --NH.sub.2, amidine, guanidine, hydroxylamine,
--C(O)NH.sub.2, secondary and tertiary amides, sulfonamides,
substituted or unsubstituted alkyl, substituted or unsubstituted
alkenyl, substituted or unsubstituted alkynyl, oxoalkyl,
oxoalkenyl, oxoalkynyl, aminoalkyl, aminoalkenyl, aminoalkynyl,
sulfoalkyl, sulfoalkenyl, sulfoalkynyl, phosphoalkyl,
phosphoalkenyl, and phosphoalkynyl groups.
[0551] In some embodiments of compounds having Formula I, the ring
structure of Recognition GroupY has the optionally substituted
structure:
##STR00054##
wherein a, b, c, d, and e are each independently carbon or
nitrogen; f is carbon, nitrogen, oxygen, or sulfur; Recognition
Group Y is attached to X and Z independently at any two ring
positions of sufficient valence; and no more than four of a, b, c,
d, e, or f are simultaneously nitrogen.
[0552] Any open valences remaining on atoms constituting the ring
structure may be filled by hydrogen or other substituents, or by
the covalent attachments to X and Z. For example, if b is carbon,
its valence may be filled by hydrogen, a substituent such as
halogen, a covalent attachment to X, or a covalent attachment to Z.
In some embodiments, a, b, c, d, and e are each carbon, while in
others, a, c, d and f are each carbon. In other embodiments, at
least one of a, b, c, d, or e is nitrogen, and in still others, f
is oxygen or sulfur. In yet another embodiment, the ring structure
of Recognition Group Y is unsubstituted. In certain embodiments,
Recognition Group Y is phenyl.
[0553] In certain embodiments of compounds of Formula I,
X-Recognition Group Y has the structure:
##STR00055##
[0554] In certain of these embodiments, v is 1, 2, 3, 4, or 5; w is
1, 2, 3, 4, or 5; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain other embodiments, v
is 1, 2 or 3 and w is 1, 2, or 3. In still other embodiments, v is
1 or 2 and w is 1 or 2.
[0555] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00056##
wherein H.sup.1 and H.sup.1' are each independently N, O, S, or
CH.sub.2; r and s are each independently 1, 2, 3, 4, or 5; and t
and t' are each independently 0, 1, 2, 3, 4, or 5. In certain of
these embodiments, H.sup.1 and H.sup.1' are each independently 0 or
CH.sub.2; r and s are each independently 1 or 2; and t and t' are
each independently 0 or 1.
[0556] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00057##
wherein H.sup.1 and H.sup.1' are each independently N, O, S, or
CH.sub.2; r and s are each independently 1, 2, 3, 4, or 5; t and t'
are each independently 0, 1, 2, 3, 4, or 5, and R.sup.b at each
occurrence is independently hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.0-6 alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain of these embodiments, H.sup.1 and
H.sup.1' are each independently 0 or CH.sub.2; r and s are each
independently 1 or 2; and t and t' are each independently 0 or
1.
[0557] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00058##
wherein H.sup.1 and H.sup.1' are each independently N, O, S, or
CH.sub.2; r and s are each independently 1, 2, 3, 4, or 5; t and t'
are each independently 0, 1, 2, 3, 4, or 5, and R.sup.b at each
occurrence is independently hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.0-6 alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain of these embodiments, H.sup.1 and
H.sup.1' are each independently 0 or CH.sub.2; r and s are each
independently 1 or 2; and t and t' are each independently 0 or
1.
[0558] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00059##
wherein H.sup.1 and H.sup.1' are each independently N, O, S, or
CH.sub.2; r and s are each independently 1, 2, 3, 4, or 5; t and t'
are each independently 0, 1, 2, 3, 4, or 5, and R.sup.b at each
occurrence is independently hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.0-6 alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain of these embodiments, H.sup.1 and
H.sup.1' are each independently 0 or CH.sub.2; r and s are each
independently 1 or 2; and t and t' are each independently 0 or
1.
[0559] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00060##
wherein H.sup.1 and H.sup.1' are each independently N, O, S, or
CH.sub.2; r and s are each independently 1, 2, 3, 4, or 5; t and t'
are each independently 0, 1, 2, 3, 4, or 5, and R.sup.b at each
occurrence is independently hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.0-6 alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain of these embodiments, H.sup.1 and
H.sup.1' are each independently 0 or CH.sub.2; r and s are each
independently 1 or 2; and t and t' are each independently 0 or
1.
[0560] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00061##
wherein H.sup.1 and H.sup.1' are each independently N, O, S, or
CH.sub.2; r and s are each independently 1, 2, 3, 4, or 5; t and t'
are each independently 0, 1, 2, 3, 4, or 5, and R.sup.b is
hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain of
these embodiments, H.sup.1 and H.sup.1' are each independently 0 or
CH.sub.2; r and s are each independently 1 or 2; and t and t' are
each independently 0 or 1.
[0561] In certain embodiments of compounds of Formula I, X has the
structure:
##STR00062##
wherein H.sup.1 and H.sup.1' are each independently N, O, S, or
CH.sub.2; r and s are each independently 1, 2, 3, 4, or 5; and t
and t' are each independently 0, 1, 2, 3, 4, or 5. In certain of
these embodiments, H.sup.1 and H.sup.1' are each independently 0 or
CH.sub.2; r and s are each independently 1 or 2; and t and t' are
each independently 0 or 1.
[0562] In certain of these embodiments of compounds of Formula I,
X-Recognition Group Y has the structure:
##STR00063##
[0563] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5, and R.sup.b at each occurrence is independently hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; w is 1; and p is 3. In some
embodiments, v is 0; t is 1, 2, or 3, w is 1; and p is 1 or 2.
[0564] In certain embodiments of compounds of Formula I,
X-Recognition Group Y has the structure:
##STR00064##
[0565] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; and R.sup.b is hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.0-6 alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain embodiments, v is 0; t is 1, 2, 3,
4, 5, or 6; r is 1 or 2; and s is 3. In some embodiments, v is 0; t
is 1, 2, or 3, r is 1; and s is 1 or 2.
[0566] In certain embodiments of compounds of Formula I,
X-Recognition Group Y has the structure:
##STR00065##
[0567] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; w is
1; and p is 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or
2; w is 1; and p is 1 or 2.
[0568] In certain embodiments of compounds of Formula I,
X-Recognition Group Y has the structure:
##STR00066##
[0569] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is
0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; and s is 3. In
some embodiments, u is 0 or 1; v is 0; t is 1, 2, or 3; r is 1; and
s is 1 or 2.
[0570] In certain embodiments of compounds of Formula I,
X-Recognition Group Y has the structure:
##STR00067##
[0571] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; w is
1; and p is 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or
2; w is 1; and p is 1 or 2.
[0572] In certain embodiments of compounds of Formula I,
X-Recognition Group Y has the structure:
##STR00068##
[0573] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is
0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; and s is 3. In
some embodiments, u is 0 or 1; v is 0; t is 1, 2, or 3, r is 1; and
s is 1 or 2.
[0574] In certain embodiments of compounds of Formula I,
X-Recognition GroupY has the structure:
##STR00069##
[0575] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; w is
1; and p is 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or
2; w is 1; and p is 1 or 2.
[0576] In certain embodiments of compounds of Formula I,
X-Recognition GroupY has the structure:
##STR00070##
[0577] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is
1 or 2; and s is 3. In some embodiments, u is 0 or 1; v is 0; t is
1, 2, or 3, r is 1; and s is 1 or 2.
[0578] In certain embodiments of compounds of Formula I,
X-Recognition GroupY has the structure:
##STR00071##
[0579] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; w is
1; and p is 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or
2; w is 1; and p is 1 or 2.
[0580] In certain embodiments of compounds of Formula I,
X-Recognition GroupY has the structure:
##STR00072##
[0581] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is
1 or 2; and s is 3. In some embodiments, u is 0 or 1; v is 0; t is
1, 2, or 3, r is 1; and s is 1 or 2.
[0582] In certain embodiments of compounds of Formula I,
X-Recognition GroupY has the structure:
##STR00073##
[0583] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; w is
1; and p is 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or
2; w is 1; and p is 1 or 2.
[0584] In certain embodiments of compounds of Formula I,
X-Recognition Group Y has the structure:
##STR00074##
[0585] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiment, u is
0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; and s is 3. In
some embodiments, u is 0 or 1; v is 0; t is 1, 2, or 3, r is 1; and
s is 1 or 2.
[0586] In compounds having Formula I wherein L has the formula
--X-Recognition Group Y--Z, the reactive group Z contains a moiety
capable of forming a covalent linkage with an amino acid in a
combining site of an antibody. For example, Z may be substituted
alkyl, substituted cycloalkyl, substituted aryl, substituted
arylalkyl, substituted heterocyclyl, or substituted
heterocyclylalkyl, wherein at least one substituent is a
1,3-diketone moiety, an acyl beta-lactam, an active ester, an
alpha-haloketone, an aldehyde, a maleimide, a lactone, an
anhydride, an alpha-haloacetamide, an amine, a hydrazide, or an
epoxide. In some such embodiments, Z is substituted alkyl.
[0587] Z may be a group that forms a reversible or irreversible
covalent bond. In some embodiments, reversible covalent bonds may
be formed using diketone Z groups such as those shown in FIG. 8.
Thus, structures A-C may form reversible covalent bonds with
reactive nucleophilic groups (e.g. lysine or cysteine side chain)
in a combining site of an antibody. R'.sub.1, R'.sub.2, R'.sub.3,
and R.sub.4 in structures A-C of FIG. 8 represent substituents
which can be C, H, N, O, P, S, halogen (F, Cl, Br, I) or a salt
thereof. These substituents also may include a group such as an
alkyl, alkenyl, alkynyl, oxoalkyl, oxoalkenyl, oxoalkynyl,
aminoalkyl, aminoalkenyl, aminoalkynyl, sulfoalkyl, sulfoalkenyl,
or sulfoalkynyl group, phosphoalkyl, phosphoalkenyl, phosphoalkynyl
group. R'.sub.2 and R'.sub.3 also could from a ring structure as
exemplified in structures B and C. X in FIG. 8 could be a
heteroatom. Other Z groups that form reversible covalent bonds
include the amidine, imine, and other reactive groups encompassed
by structure G of FIG. 8. FIG. 9 includes the structures of other
linker reactive groups that form reversible covalent bonds, e.g.,
structures B, G, H, and, where X is not a leaving group, E and
F.
[0588] Z reactive groups that form an irreversible covalent bond
with a combining site of an antibody include structures D-G in FIG.
8 (e.g., when G is an imidate) and structures A, C and D of FIG. 9.
When X is a leaving group, structures E and F of FIG. 9 may also
form irreversible covalent bonds. Such structures are useful for
irreversibly attaching a targeting agent-linker to a reactive
nucleophilic group to a combining site of an antibody.
[0589] In other such embodiments, Z is a 1,3-diketone moiety. In
still other such embodiments, Z is alkyl substituted by a
1,3-diketone moiety. In certain embodiments, Z has the
structure:
##STR00075##
wherein q=0-5. In certain other embodiments, Z has the
structure:
##STR00076##
[0590] One linker for use in AA targeting compounds and for
preparing AA targeting agent-linker compounds includes a
1,3-diketone reactive group as Z. In certain embodiments of Formula
I, L has the structure:
##STR00077##
[0591] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0592] In certain embodiments of Formula I, Linker has the
structure:
##STR00078##
[0593] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0594] In certain embodiments of Formula I, Linker has the
structure:
##STR00079##
[0595] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0596] In certain embodiments of Formula I, Linker has the
structure:
##STR00080##
[0597] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0598] In certain embodiments of Formula I, Linker has the
structure:
##STR00081##
[0599] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0600] In certain embodiments of Formula I, Linker has the
structure:
##STR00082##
[0601] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is 0;
t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2, or
3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1 or
2; and q is 2 or 3.
[0602] In certain embodiments of Formula I, Linker has the
structure:
##STR00083##
[0603] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b
at each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2,
or 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or 2; w is
1; p is 1 or 2; and q is 1 or 2.
[0604] In certain embodiments of Formula I, Linker has the
structure:
##STR00084##
[0605] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b
at each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1 or 2; w is 1; p is 1 or
2; and q is 1 or 2.
[0606] In certain embodiments of Formula I, Linker has the
structure:
##STR00085##
[0607] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, or 3; and R.sup.b at
each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1 or 2; w is 1; p is 1 or
2; and q is 2 or 3.
[0608] In certain embodiments of Formula I, Linker has the
structure:
##STR00086##
[0609] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and
R.sup.b is hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain
embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, u is 0 or 1; v
is 0; t is 1 or 2; w is 1; p is 1 or 2; and q is 1 or 2.
[0610] In certain embodiments of Formula I, Linker has the
structure:
##STR00087##
[0611] In certain of these embodiments u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and
R.sup.b is hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain
embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, u is 0 or 1; v
is 0; t is 1, 2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0612] In certain embodiments of Formula I, Linker has the
structure:
##STR00088##
[0613] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, or 3; and R.sup.b is
hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain
embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, u is 0 or 1; v
is 0; t is 1, 2, or 3, r is 1; s is 1 or 2; and q is 2 or 3.
[0614] In certain embodiments of Formula I, Linker has the
structure:
##STR00089##
[0615] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b
at each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.m alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0; v is 0; t is
1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2, or 3. In
some embodiments, u is 0 or 1; v is 0; t is 1 or 2; w is 1; p is 1
or 2; and q is 1 or 2.
[0616] In certain embodiments of Formula I, Linker has the
structure:
##STR00090##
[0617] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b
at each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2,
or 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or 2; w is
1; p is 1 or 2; and q is 1 or 2.
[0618] In certain embodiments of Formula I, Linker has the
structure:
##STR00091##
[0619] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, or 3; and R.sup.b at
each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2,
or 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or 2; w is
1; p is 1 or 2; and q is 2 or 3.
[0620] In certain embodiments of Formula I, Linker has the
structure:
##STR00092##
[0621] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and
R.sup.b is hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain
embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, u is 0 or 1; v
is 0; t is 1, 2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0622] In certain embodiments of Formula I, Linker has the
structure:
##STR00093##
[0623] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and
R.sup.b is hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain
embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, u is 0 or 1; v
is 0; t is 1, 2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0624] In certain embodiments of Formula I, Linker has the
structure:
##STR00094##
[0625] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, or 3; and R.sup.b is
hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain
embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, u is 0 or 1; v
is 0; t is 1, 2, or 3, r is 1; s is 1 or 2; and q is 2 or 3.
[0626] In certain embodiments of Formula I, Linker has the
structure:
##STR00095##
[0627] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b
at each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2,
or 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or 2; w is
1; p is 1 or 2; and q is 1 or 2.
[0628] In certain embodiments of Formula I, Linker has the
structure:
##STR00096##
[0629] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b
at each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2,
or 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or 2; w is
1; p is 1 or 2; and q is 1 or 2.
[0630] In certain embodiments of Formula I, Linker has the
structure:
##STR00097##
[0631] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, or 3; and R.sup.b at
each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2,
or 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or 2; w is
1; p is 1 or 2; and q is 2 or 3.
[0632] In certain embodiments of Formula I, Linker has the
structure:
##STR00098##
[0633] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and
R.sup.b at each occurrence is independently hydrogen, substituted
or unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0,
1, 2, or 3. In some embodiments, u is 0 or 1; v is 0; t is 1, 2, or
3, r is 1; s is 1 or 2; and q is 1 or 2.
[0634] In certain embodiments of Formula I, Linker has the
structure:
##STR00099##
[0635] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and
R.sup.b at each occurrence is independently hydrogen, substituted
or unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0,
1, 2, or 3. In some embodiments, u is 0 or 1; v is 0; t is 1, 2, or
3, r is 1; s is 1 or 2; and q is 1 or 2.
[0636] In certain embodiments of Formula I, Linker has the
structure:
##STR00100##
[0637] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, or 3; and R.sup.b at
each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0,
1, 2, or 3. In some embodiments, u is 0 or 1; v is 0; t is 1, 2, or
3, r is 1; s is 1 or 2; and q is 2 or 3.
[0638] In certain embodiments of Formula I, Linker has the
structure:
##STR00101##
[0639] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b
at each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2,
or 3. In still some embodiments, u is 0 or 1; v is 0; t is 1 or 2;
w is 1; p is 1 or 2; and q is 1 or 2.
[0640] In certain embodiments of Formula I, Linker has the
structure:
##STR00102##
[0641] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b
at each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2,
or 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or 2; w is
1; p is 1 or 2; and q is 1 or 2.
[0642] In certain embodiments of Formula I, Linker has the
structure:
##STR00103##
[0643] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, or 3; and R.sup.b at
each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2,
or 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or 2; w is
1; p is 1 or 2; and q is 2 or 3.
[0644] In certain embodiments of Formula I, Linker has the
structure:
##STR00104##
[0645] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and
R.sup.b at each occurrence is independently hydrogen, substituted
or unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0,
1, 2, or 3. In some embodiments, u is 0 or 1; v is 0; t is 1, 2, or
3, r is 1; s is 1 or 2; and q is 1 or 2.
[0646] In certain embodiments of Formula I, Linker has the
structure:
##STR00105##
[0647] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and
R.sup.b at each occurrence is independently hydrogen, substituted
or unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0,
1, 2, or 3. In some embodiments, u is 0 or 1; v is 0; t is 1, 2, or
3, r is 1; s is 1 or 2; and q is 1 or 2.
[0648] In certain embodiments of Formula I, Linker has the
structure:
##STR00106##
[0649] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, or 3; and R.sup.b at
each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0,
1, 2, or 3. In some embodiments, u is 0 or 1; v is 0; t is 1, 2, or
3, r is 1; s is 1 or 2; and q is 2 or 3.
[0650] In certain embodiments of Formula I, Linker has the
structure:
##STR00107##
[0651] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b
at each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.m alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0; v is 0; t is
1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2, or 3. In
still other embodiments, u is 0 or 1; v is 0; t is 1 or 2; w is 1;
p is 1 or 2; and q is 1 or 2.
[0652] In certain embodiments of Formula I, Linker has the
structure:
##STR00108##
[0653] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b
at each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0;
v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2,
or 3. In some embodiments, u is 0 or 1; v is 0; t is 1 or 2; w is
1; p is 1 or 2; and q is 1 or 2.
[0654] In certain embodiments of Formula I, Linker has the
structure:
##STR00109##
[0655] In certain of these embodiments, u is 0, 1, 2, 3, 4, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4,
or 5; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, or 3; and R.sup.b at
each occurrence is independently hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.m alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain embodiments, u is 0; v is 0; t is
1, 2, 3, 4, 5, or 6; w is 1; p is 3; and q is 0, 1, 2, or 3. In
some embodiments, u is 0 or 1; v is 0; t is 1 or 2; w is 1; p is 1
or 2; and q is 2 or 3.
[0656] In certain embodiments of Formula I, Linker has the
structure:
##STR00110##
[0657] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and
R.sup.b is hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain
embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, u is 0 or 1; v
is 0; t is 1, 2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0658] In certain embodiments of Formula I, Linker has the
structure:
##STR00111##
[0659] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and
R.sup.b is hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain
embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, u is 0 or 1; v
is 0; t is 1, 2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0660] In certain embodiments of Formula I, Linker has the
structure:
##STR00112##
[0661] In certain of these embodiments, u is 0, 1, 2, 3, 5, or 5; v
is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4,
or 5; s is 0, 1, 2, 3, 4, or 5; q is 0, 1, 2, or 3; and R.sup.b is
hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain
embodiments, u is 0; v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, u is 0 or 1; v
is 0; t is 1, 2, or 3, r is 1; s is 1 or 2; and q is 2 or 3.
[0662] As used herein, "AA.sub.1-AA.sub.2-AA.sub.n" refers to an AA
targeting agent wherein "AA.sub.1" is the first amino acid in the
AA targeting agent sequence, as measured from the N-terminus,
"AA.sub.2" is the second amino acid in the AA targeting agent
sequence, as measured from the N-terminus, and "AA.sub.n" is the
n.sup.th amino acid in the AA targeting agent sequence, as measured
from the N-terminus.
[0663] Certain embodiments in accordance with Formula I have the
structure:
##STR00113##
[0664] In certain of these embodiments, v is 1, 2, 3, 4, or 5; w is
1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is
hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain
embodiments, v is 1, 2, or 3; w is 1, 2, or 3; and q is 0, 1, 2, or
3. In some embodiments, v is 1 or 2; w is 1 or 2; and q is 1 or
2.
[0665] Certain embodiments in accordance with Formula I have the
structure:
##STR00114##
[0666] In certain of these embodiments, v is 1, 2, 3, 4, or 5; w is
1, 2, 3, 4, or 5; q is 0, 1, 2, or 3; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
1, 2, or 3; w is 1, 2, or 3; and q is 0, 1, 2, 3. In some
embodiments, v is 1 or 2; w is 1 or 2; and q is 2 or 3.
[0667] Certain embodiments in accordance with Formula I have the
structure:
##STR00115##
[0668] In certain of these embodiments, v is 1, 2, 3, 4, or 5; w is
1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is
hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain
embodiments, v is 1, 2, or 3; w is 1, 2, or 3; and q is 0, 1, 2, 3.
In some embodiments, v is 1 or 2; w is 1 or 2; and q is 2 or 3.
[0669] Certain embodiments in accordance with Formula I have the
structure:
##STR00116##
[0670] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0671] Certain embodiments in accordance with Formula I have the
structure:
##STR00117##
[0672] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0673] Certain embodiments in accordance with Formula I have the
structure:
##STR00118##
[0674] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0, 1, 2, 3, 4, or 5; t is 1, 2, 3, 4, 5,
or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or 5; and q is 0, 1,
2, or 3. In certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6;
w is 1; and p is 3, and in some embodiments, v is 0; t is 1 or 2; w
is 1; p is 1 or 2; and q is 2 or 3.
[0675] Certain embodiments in accordance with Formula I have the
structure:
##STR00119##
[0676] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0677] Certain embodiments in accordance with Formula I have the
structure:
##STR00120##
[0678] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.m alkyl, or substituted or
unsubstituted aryl-C.sub.m alkyl. In certain embodiments, v is 0; t
is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2, or
3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1 or
2; and q is 1 or 2.
[0679] Certain embodiments in accordance with Formula I have the
structure:
##STR00121##
[0680] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.m alkyl, or substituted or unsubstituted
aryl-C.sub.m alkyl. In certain embodiments, v is 0; t is 1, 2, 3,
4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2, or 3. In some
embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1 or 2; and q is
2 or 3.
[0681] Certain embodiments in accordance with Formula I have the
structure:
##STR00122##
[0682] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-C.sub.m alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0683] Certain embodiments in accordance with Formula I have the
structure:
##STR00123##
[0684] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0685] Certain embodiments in accordance with Formula I have the
structure:
##STR00124##
[0686] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0687] Certain embodiments in accordance with Formula I have the
structure:
##STR00125##
[0688] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0689] Certain embodiments in accordance with Formula I have the
structure:
##STR00126##
[0690] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0691] Certain embodiments in accordance with Formula I have the
structure:
##STR00127##
[0692] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is 0;
t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2, or
3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1 or
2; and q is 2 or 3.
[0693] Certain embodiments in accordance with Formula I have the
structure:
##STR00128##
[0694] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0695] Certain embodiments in accordance with Formula I have the
structure:
##STR00129##
[0696] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0697] Certain embodiments in accordance with Formula I have the
structure:
##STR00130##
[0698] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0699] Certain embodiments in accordance with Formula I have the
structure:
##STR00131##
[0700] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is
1, 2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0701] Certain embodiments in accordance with Formula I have the
structure:
##STR00132##
[0702] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.m alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is
1, 2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0703] Certain embodiments in accordance with Formula I have the
structure:
##STR00133##
[0704] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-C.sub.m alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is
1, 2, or 3, r is 1; s is 1 or 2; and q is 2 or 3.
[0705] Certain embodiments in accordance with Formula I have the
structure:
##STR00134##
[0706] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0707] Certain embodiments in accordance with Formula I have the
structure:
##STR00135##
[0708] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0709] Certain embodiments in accordance with Formula I have the
structure:
##STR00136##
[0710] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0711] Certain embodiments in accordance with Formula I have the
structure:
##STR00137##
[0712] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is
1, 2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0713] Certain embodiments in accordance with Formula I have the
structure:
##STR00138##
[0714] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is
1, 2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0715] Certain embodiments in accordance with Formula I have the
structure:
##STR00139##
[0716] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-O.sub.06 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is
1, 2, or 3, r is 1; s is 1 or 2; and q is 2 or 3.
[0717] Certain embodiments in accordance with Formula I have the
structure:
##STR00140##
[0718] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.m alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0719] Certain embodiments in accordance with Formula I have the
structure:
##STR00141##
[0720] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-O.sub.06 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0721] Certain embodiments in accordance with Formula I have the
structure:
##STR00142##
[0722] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0723] Certain embodiments in accordance with Formula I have the
structure:
##STR00143##
[0724] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0725] Certain embodiments in accordance with Formula I have the
structure:
##STR00144##
[0726] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0727] Certain embodiments in accordance with Formula I have the
structure:
##STR00145##
[0728] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is 0;
t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2, or
3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1 or
2; and q is 2 or 3.
##STR00146##
as used herein refers to an AA targeting agent wherein "AA.sub.1"
is the first amino acid in an AA targeting agent sequence as
measured from the N-terminus, "AA.sub.2" is the second amino acid
in an AA targeting agent sequence as measured from the N-terminus,
and "AA.sub.n" is the n.sup.th amino acid in an AA targeting agent
sequence as measured from the N-terminus. The targeting agent
further comprises a Lys residue at arbitrary position m+1 as
measured from the N-terminus. It will be appreciated that in
addition to linking to a Lys side chain in the body of an AA
targeting agent, it is also possible to link to a Lys side chain on
the N-terminus or C-terminus of an AA targeting agent.
[0729] Certain embodiments in accordance with Formula I have the
structure:
##STR00147##
[0730] In certain of these embodiments, v is 1, 2, 3, 4, or 5; w is
1, 2, 3, 4, 5, or 6; q is 1, 2, 3, 4, or 5; and R.sub.b at each
occurrence is independently hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.0-6 alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain embodiments, v is 1, 2 or 3; w is
1, 2 or 3; and q is 1, 2, or 3. In some embodiments, v is 1 or 2; w
is 1 or 2; and q is 2, or 3.
[0731] Certain embodiments in accordance with Formula I have the
structure:
##STR00148##
[0732] In certain of these embodiments, v is 1, 2, 3, 4, or 5; w is
1, 2, 3, 4, 5, or 6; q is 1, 2, 3, 4, or 5; and R.sub.b at each
occurrence is independently hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.m alkyl, or substituted or unsubstituted
aryl-O.sub.06 alkyl. In certain embodiments, v is 1, 2 or 3; w is
1, 2 or 3; and q is 1, 2, or 3. In some embodiments, v is 1 or 2; w
is 1 or 2; and q is 2, or 3.
[0733] Certain embodiments in accordance with Formula I have the
structure:
##STR00149##
[0734] In certain of these embodiments, v is 1, 2, 3, 4, or 5; w is
1, 2, 3, 4, 5, or 6; q is 1, 2, 3, 4, or 5; and R.sub.b at each
occurrence is independently hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.m alkyl, or substituted or unsubstituted
aryl-C.sub.m alkyl. In certain embodiments, v is 1, 2 or 3; w is 1,
2 or 3; and q is 1, 2, or 3. In some embodiments, v is 1 or 2; w is
1 or 2; and q is 2, or 3.
[0735] Certain embodiments in accordance with Formula I have the
structure:
##STR00150##
[0736] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0737] Certain embodiments in accordance with Formula I have the
structure:
##STR00151##
[0738] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0739] Certain embodiments in accordance with Formula I have the
structure:
##STR00152##
[0740] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0741] Certain embodiments in accordance with Formula I have the
structure:
##STR00153##
[0742] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0743] Certain embodiments in accordance with Formula I have the
structure:
##STR00154##
[0744] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0745] Certain embodiments in accordance with Formula I have the
structure:
##STR00155##
[0746] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is 0;
t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2, or
3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1 or
2; and q is 2 or 3.
[0747] Certain embodiments in accordance with Formula I have the
structure:
##STR00156##
[0748] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0749] Certain embodiments in accordance with Formula I have the
structure:
##STR00157##
[0750] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0751] Certain embodiments in accordance with Formula I have the
structure:
##STR00158##
[0752] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0753] Certain embodiments in accordance with Formula I have the
structure:
##STR00159##
[0754] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0755] Certain embodiments in accordance with Formula I have the
structure:
##STR00160##
[0756] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0757] Certain embodiments in accordance with Formula I have the
structure:
##STR00161##
[0758] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is 0;
t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2, or
3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1 or
2; and q is 2 or 3.
[0759] Certain embodiments in accordance with Formula I have the
structure:
##STR00162##
[0760] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0761] Certain embodiments in accordance with Formula I have the
structure:
##STR00163##
[0762] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiment v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0763] Certain embodiments in accordance with Formula I have the
structure:
##STR00164##
[0764] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-C.sub.m alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0765] Certain embodiments in accordance with Formula I have the
structure:
##STR00165##
[0766] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-C.sub.m alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is
1, 2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0767] Certain embodiments in accordance with Formula I have the
structure:
##STR00166##
[0768] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is
1, 2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0769] Certain embodiments in accordance with Formula I have the
structure:
##STR00167##
[0770] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is
1, 2, or 3, r is 1; s is 1 or 2; and q is 2 or 3.
[0771] The administration of an AA targeting compound to an
immunocompetent individual may result in the production of
antibodies against the conjugate. Such antibodies may be directed
to the variable region, including the antibody idiotype, as well as
to the targeting agent or any linker used to conjugate the
targeting agent to the antibody. Reducing the immunogenicity of an
AA targeting compound can be accomplished by methods well known in
the art, such as by attaching long chain polyethylene glycol
(PEG)-based spacers and the like to the AA targeting compound. Long
chain PEG and other polymers are known for their ability to mask
foreign epitopes, resulting in the reduced immunogenicity of
therapeutic proteins that display foreign epitopes (N. V. Katre, J.
Immunol. 144:209-213 (1990); G. E. Francis et al., Int. J. Hematol.
68:1-18 (1998). Alternatively, or in addition, the individual
administered the antibody-AA targeting agent conjugate may be
administered an immunosuppressant such as cyclosporin A, anti-CD3
antibody, and the like.
[0772] In one embodiment, an AA targeting compound is as shown by
Formula II, and includes stereoisomers, tautomers, solvates,
prodrugs, and pharmaceutically acceptable salts thereof.
Antibody-Linker'-[AA targeting agent] (II)
[0773] In compounds of Formula II, [AA targeting agent] is defined
as in Formula I. Linker' is a linker moiety linking an antibody to
the targeting agent and having the formula --X-Recognition Group
Y--Z'--. In compounds of Formula II, X and Y are defined as in
Formula I, and Antibody is an antibody as defined herein. FIGS. 10
and 11, respectively, illustrate the addition mechanism of a
reactive, nucleophilic side chain in a combining site of an
antibody to the Z moieties illustrated in FIGS. 8 and 9.
[0774] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00168##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0775] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00169##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0776] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00170##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0777] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00171##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0778] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00172##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0779] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00173##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0780] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00174##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0781] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00175##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0782] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00176##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0783] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00177##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0784] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00178##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0785] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00179##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0786] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00180##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0787] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00181##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0788] In certain embodiments, wherein Antibody is an aldolase
catalytic antibody, Z'-- Antibody has the structure:
##STR00182##
wherein HN-Antibody refers to an arbitrary side chain in the
combining site of an antibody bearing an amino group.
[0789] In compounds having Formula II, Z' is an attachment moiety
comprising a covalent bond and 0-20 carbon atoms to which the
Antibody is attached. This is shown below for the case where the
linker has a diketone moiety as the reactive group and linkage
occurs with the side chain amino group of a lysine residue in the
antibody combining site. The Antibody is shown schematically as
bivalent with a reactive amino acid side chain for each combining
site indicated.
##STR00183##
[0790] Another embodiment shown below is for the case where the
linker has a beta lactam moiety as the reactive group and linkage
occurs with the side chain amino group of a lysine residue in the
antibody combining site. The Antibody is shown schematically as
bivalent with a reactive amino acid side chain for each combining
site indicated.
##STR00184##
[0791] Certain embodiments in accordance with Formula II have the
structure:
##STR00185##
[0792] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-C.sub.m alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0793] Certain embodiments in accordance with Formula II have the
structure:
##STR00186##
[0794] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-C.sub.m alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0795] Certain embodiments in accordance with Formula II have the
structure:
##STR00187##
[0796] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; and p is 1, 2, 3, 4,
or 5; and R.sup.b at each occurrence is independently hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; w is 1; and p is 3. In some
embodiments, v is 0; t is 1 or 2; w is 1; and p is 1 or 2.
[0797] Certain embodiments in accordance with Formula II have the
structure:
##STR00188##
[0798] In certain of these embodiments, v is 1, 2, 3, 4, or 5; w is
1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is
hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain
embodiments, v is 1, 2, or 3; w is 1, 2, or 3; and q is 0, 1, 2, 3.
In some embodiments, v is 1 or 2; w is 1 or 2; and q is 1 or 2.
[0799] Certain embodiments in accordance with Formula II have the
structure:
##STR00189##
[0800] In certain of these embodiments, v is 1, 2, 3, 4, or 5; w is
1, 2, 3, 4, or 5; q is 0, 1, 2, or 3; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
1, 2, or 3; w is 1, 2, or 3; and q is 0, 1, 2, 3. some embodiments,
v is 1 or 2; w is 1 or 2; and q is 2 or 3.
[0801] Certain embodiments in accordance with Formula II have the
structure:
##STR00190##
[0802] In certain of these embodiments, v is 1, 2, 3, 4, or 5; w is
1, 2, 3, 4, or 5; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is 1,
2, or 3; and w is 1, 2, or 3. In some embodiments, v is 1 or 2 and
w is 1 or 2.
[0803] Certain embodiments in accordance with Formula II have the
structure:
##STR00191##
[0804] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3 and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0805] Certain embodiments in accordance with Formula II have the
structure:
##STR00192##
[0806] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is 0;
t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3 and q is 0, 1, 2, or
3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1 or
2; and q is 2 or 3.
[0807] Certain embodiments in accordance with Formula II have the
structure:
##STR00193##
[0808] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; and R.sup.b is hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.m alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain embodiments, v is 0; t is 1, 2, 3,
4, 5, or 6; r is 1 or 2; and s is 3. In some embodiments, v is 0; t
is 1, 2, or 3, r is 1; and s is 1 or 2.
[0809] Certain embodiments in accordance with Formula II have the
structure:
##STR00194##
[0810] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0811] Certain embodiments in accordance with Formula II have the
structure:
##STR00195##
[0812] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0813] Certain embodiments in accordance with Formula II have the
structure:
##STR00196##
[0814] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; and R.sup.b at each occurrence is independently hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; w is 1; and p is 3. In some
embodiments, v is 0; t is 1 or 2; w is 1; and p is 1 or 2.
[0815] Certain embodiments in accordance with Formula II have the
structure:
##STR00197##
[0816] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3 and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0817] Certain embodiments in accordance with Formula II have the
structure:
##STR00198##
[0818] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is 0;
t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3 and q is 0, 1, 2, or
3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1 or
2; and q is 2 or 3.
[0819] Certain embodiments in accordance with Formula II have the
structure:
##STR00199##
[0820] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; and R.sup.b is hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.0-6 alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain embodiments, v is 0; t is 1, 2, 3,
4, 5, or 6; r is 1 or 2; and s is 3. In some embodiments, v is 0; t
is 1, 2, or 3, r is 1; and s is 1 or 2.
[0821] Certain embodiments in accordance with Formula II have the
structure:
##STR00200##
[0822] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0823] Certain embodiments in accordance with Formula II have the
structure:
##STR00201##
[0824] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0825] Certain embodiments in accordance with Formula II have the
structure:
##STR00202##
[0826] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; and R.sup.b at each occurrence are independently hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.m alkyl, or substituted or
unsubstituted aryl-C.sub.m alkyl. In certain embodiments, v is 0; t
is 1, 2, 3, 4, 5, or 6; w is 1; and p is 3. In some embodiments, v
is 0; t is 1 or 2; w is 1; and p is 1 or 2.
[0827] Certain embodiments in accordance with Formula II have the
structure:
##STR00203##
[0828] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-C.sub.m alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3 and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1,
2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0829] Certain embodiments in accordance with Formula II have the
structure:
##STR00204##
[0830] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-O.sub.06 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3 and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1,
2, or 3, r is 1; s is 1 or 2; and q is 2 or 3.
[0831] Certain embodiments in accordance with Formula II have the
structure:
##STR00205##
[0832] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; and R.sup.b at each occurrence is independently hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; and s is 3. In some
embodiments, v is 0; t is 1, 2, or 3, r is 1; and s is 1 or 2.
[0833] Certain embodiments in accordance with Formula II have the
structure:
##STR00206##
[0834] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0835] Certain embodiments in accordance with Formula II have the
structure:
##STR00207##
[0836] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0837] Certain embodiments in accordance with Formula II have the
structure:
##STR00208##
[0838] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; and R.sup.b at each occurrence is independently hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; w is 1; and p is 3. In some
embodiments, v is 0; t is 1 or 2; w is 1; and p is 1 or 2.
[0839] Certain embodiments in accordance with Formula II have the
structure:
##STR00209##
[0840] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3 and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0841] Certain embodiments in accordance with Formula II have the
structure:
##STR00210##
[0842] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.m alkyl, or substituted or unsubstituted
aryl-C.sub.m alkyl. In certain embodiments, v is 0; t is 1, 2, 3,
4, 5, or 6; r is 1 or 2; s is 3 and q is 0, 1, 2, or 3. In some
embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1 or 2; and q is
2 or 3.
[0843] Certain embodiments in accordance with Formula II have the
structure:
##STR00211##
[0844] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; and R.sup.b is hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.m alkyl, or substituted or unsubstituted
aryl-C.sub.m alkyl. In certain embodiments, v is 0; t is 1, 2, 3,
4, 5, or 6; r is 1 or 2; and s is 3. In some embodiments, v is 0; t
is 1, 2, or 3, r is 1; and s is 1 or 2.
[0845] Certain embodiments in accordance with Formula II have the
structure:
##STR00212##
[0846] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-C.sub.m alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0847] Certain embodiments in accordance with Formula II have the
structure:
##STR00213##
[0848] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0849] Certain embodiments in accordance with Formula II have the
structure:
##STR00214##
[0850] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; and R.sup.b at each occurrence is independently hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; w is 1; and p is 3. In some
embodiments, v is 0; t is 1 or 2; w is 1; and p is 1 or 2.
[0851] Certain embodiments in accordance with Formula II have the
structure:
##STR00215##
[0852] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3 and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1,
2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0853] Certain embodiments in accordance with Formula II have the
structure:
##STR00216##
[0854] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3 and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1,
2, or 3, r is 1; s is 1 or 2; and q is 2 or 3.
[0855] Certain embodiments in accordance with Formula II have the
structure:
##STR00217##
[0856] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; and R.sup.b at each occurrence is independently hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; and s is 3. In some
embodiments, v is 0; t is 1, 2, or 3, r is 1; and s is 1 or 2.
[0857] Certain embodiments in accordance with Formula II have the
structure:
##STR00218##
[0858] In certain of these embodiments, v is 1, 2, 3, 4, or 5; w is
1, 2, 3, 4, 5, or 6; q is 1, 2, 3, 4, or 5; and R.sub.b at each
occurrence is independently hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.0-6 alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain embodiments, v is 1, 2 or 3; w is
1, 2 or 3; and q is 1, 2, or 3. In some embodiments, v is 1 or 2; w
is 1 or 2; and q is 2, or 3.
[0859] Certain embodiments in accordance with Formula II have the
structure:
##STR00219##
[0860] In certain of these embodiments, v is 1, 2, 3, 4, or 5; and
w is 1, 2, 3, 4, 5; and R.sub.b at each occurrence is independently
hydrogen, substituted or unsubstituted C.sub.1-10 alkyl,
substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl,
or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In certain
embodiments, v is 1, 2 or 3; and w is 1, 2 or 3. In some
embodiments, v is 1 or 2; and w is 1 or 2.
[0861] Certain embodiments in accordance with Formula II have the
structure:
##STR00220##
[0862] In certain of these embodiments, v is 1, 2, 3, 4, or 5; w is
1, 2, 3, 4, 5, or 6; q is 1, 2, 3, 4, or 5; and R.sub.b at each
occurrence is independently hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.0-6 alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain embodiments, v is 1, 2 or 3; w is
1, 2 or 3; and q is 1, 2, or 3. In some embodiments, v is 1 or 2; w
is 1 or 2; and q is 2, or 3.
[0863] Certain embodiments in accordance with Formula II have the
structure:
##STR00221##
[0864] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0865] Certain embodiments in accordance with Formula II have the
structure:
##STR00222##
[0866] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0867] Certain embodiments in accordance with Formula II have the
structure:
##STR00223##
[0868] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; and R.sup.b at each occurrence is independently hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; w is 1; and p is 3. In some
embodiments, v is 0; t is 1 or 2; w is 1; and p is 1 or 2.
[0869] Certain embodiments in accordance with Formula II have the
structure:
##STR00224##
[0870] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0871] Certain embodiments in accordance with Formula II have the
structure:
##STR00225##
[0872] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is 0;
t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2, or
3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1 or
2; and q is 2 or 3.
[0873] Certain embodiments in accordance with Formula II have the
structure:
##STR00226##
[0874] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; and R.sup.b is hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.m alkyl, or substituted or unsubstituted
aryl-C.sub.m alkyl. In certain embodiments, v is 0; t is 1, 2, 3,
4, 5, or 6; r is 1 or 2; and s is 3. In some embodiments, v is 0; t
is 1, 2, or 3, r is 1; and s is 1 or 2.
[0875] Certain embodiments in accordance with Formula II have the
structure:
##STR00227##
[0876] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.m
alkyl, or substituted or unsubstituted aryl-C.sub.m alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0877] Certain embodiments in accordance with Formula II have the
structure:
##STR00228##
[0878] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0879] Certain embodiments in accordance with Formula II have the
structure:
##STR00229##
[0880] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; and R.sup.b at each occurrence is independently hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; w is 1; and p is 3. In some
embodiments, v is 0; t is 1 or 2; w is 1; and p is 1 or 2.
[0881] Certain embodiments in accordance with Formula II have the
structure:
##STR00230##
[0882] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b is hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2,
or 3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1
or 2; and q is 1 or 2.
[0883] Certain embodiments in accordance with Formula II have the
structure:
##STR00231##
[0884] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b is hydrogen, substituted or
unsubstituted C.sub.1-10 alkyl, substituted or unsubstituted
C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted or
unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is 0;
t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; s is 3; and q is 0, 1, 2, or
3. In some embodiments, v is 0; t is 1, 2, or 3, r is 1; s is 1 or
2; and q is 2 or 3.
[0885] Certain embodiments in accordance with Formula II have the
structure:
##STR00232##
[0886] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; and R.sup.b is hydrogen, substituted or unsubstituted
C.sub.1-10 alkyl, substituted or unsubstituted C.sub.3-7
cycloalkyl-C.sub.0-6 alkyl, or substituted or unsubstituted
aryl-C.sub.0-6 alkyl. In certain embodiments, v is 0; t is 1, 2, 3,
4, 5, or 6; r is 1 or 2; and s is 3. In some embodiments, v is 0; t
is 1, 2, or 3, r is 1; and s is 1 or 2.
[0887] Certain embodiments in accordance with Formula II have the
structure:
##STR00233##
[0888] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 1 or 2.
[0889] Certain embodiments in accordance with Formula II have the
structure:
##STR00234##
[0890] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; w is 1; p is
3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is 1 or
2; w is 1; p is 1 or 2; and q is 2 or 3.
[0891] Certain embodiments in accordance with Formula II have the
structure:
##STR00235##
[0892] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; w is 1, 2, 3, 4, or 5; p is 1, 2, 3, 4, or
5; and R.sup.b at each occurrence is independently hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; w is 1; and p is 3. In some
embodiments, v is 0; t is 1 or 2; w is 1; and p is 1 or 2.
[0893] Certain embodiments in accordance with Formula II have the
structure:
##STR00236##
[0894] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, 3, 4, or 5; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is
1, 2, or 3, r is 1; s is 1 or 2; and q is 1 or 2.
[0895] Certain embodiments in accordance with Formula II have the
structure:
##STR00237##
[0896] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; q is 0, 1, 2, or 3; and R.sup.b at each occurrence is
independently hydrogen, substituted or unsubstituted C.sub.1-10
alkyl, substituted or unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6
alkyl, or substituted or unsubstituted aryl-C.sub.0-6 alkyl. In
certain embodiments, v is 0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2;
s is 3; and q is 0, 1, 2, or 3. In some embodiments, v is 0; t is
1, 2, or 3, r is 1; s is 1 or 2; and q is 2 or 3.
[0897] Certain embodiments in accordance with Formula II have the
structure:
##STR00238##
[0898] In certain of these embodiments, v is 0, 1, 2, 3, 4, or 5; t
is 1, 2, 3, 4, 5, or 6; r is 1, 2, 3, 4, or 5; s is 0, 1, 2, 3, 4,
or 5; and R.sup.b at each occurrence is independently hydrogen,
substituted or unsubstituted C.sub.1-10 alkyl, substituted or
unsubstituted C.sub.3-7 cycloalkyl-C.sub.0-6 alkyl, or substituted
or unsubstituted aryl-C.sub.0-6 alkyl. In certain embodiments, v is
0; t is 1, 2, 3, 4, 5, or 6; r is 1 or 2; and s is 3. In some
embodiments, v is 0; t is 1, 2, or 3, r is 1; and s is 1 or 2.
[0899] Alternatively, the linker may have an amine or hydrazide as
the reactive group and the Antibody may be engineered to have a
diketone moiety. An unnatural diketone-containing amino acid may be
readily incorporated into an antibody combining site using
techniques well known in the art; proteins containing unnatural
amino acids have been produced in yeast and bacteria. See, e.g., J.
W. Chin et al., Science 301:964-966 (2003); L. Wang et al., Science
292:498-500 (2001); J. W. Chin et al., J. Am. Chem. Soc.
124:9026-9027 (2002); L. Wang, et al., J. Am. Chem. Soc.
124:1836-1837 (2002); J. W. Chin and P. G. Schultz, Chembiochem.
3:1135-1137 (2002); J. W. Chin et al., Proc. Natl. Acad. Sci.
U.S.A. 99:11020-11024 (2002); L. Wang and P. G. Schultz, Chem.
Commun. (1):1-11 (2002); Z. Zhang et al., Angew. Chem. Int. Ed.
Engl. 41:2840-2842 (2002); L. Wang, Proc. Natl. Acad. Sci. U.S.A.
100:56-61 (2003). Thus, for example, to insert an unnatural amino
acid containing a diketone moiety into the yeast Saccharomyces
cerevisiae requires the addition of new components to the protein
biosynthetic machinery including a unique codon, tRNA, and
aminoacyl-tRNA synthetase (aa RS). For example, the amber
suppressor tyrosyl-tRNA synthetase (TyrRS)-tRNA.sub.CUA pair from
E. coli may be used as reported for eukaryotes in J. W. Chin et
al., Science 301:964-966 (2003). The amber codon is used to code
for the unnatural amino acid of interest. Libraries of mutant TyrRS
and tRNA.sub.CUA may then be produced and selected for those
aaRS-tRNA.sub.CUA pairs in which the TyrRS charges the tRNA.sub.CUA
with the unnatural amino acid of interest, e.g., the
diketone-containing amino acid. Subsequently, antibodies
incorporating the diketone-containing amino acid may be produced by
cloning and expressing a gene containing the amber codon at one or
more antibody combining sites.
[0900] In some embodiments of compounds of Formula II, the Antibody
is a full length antibody. In other embodiments, the Antibody is
Fab, Fab' F(ab').sub.2, Fv, V.sub.H, V.sub.L, or scFv. In certain
embodiments, the Antibody is a human antibody, humanized antibody
or chimeric human antibody. In certain embodiments, the Antibody is
a catalytic antibody. In one embodiment, the Antibody is a
humanized version of a murine 38c2 comprising a constant region
from a human IgG, IgA, IgM, IgD, or IgE antibody. In another
embodiment, Antibody is a chimeric antibody comprising the variable
region from murine 38c2 and a constant region from a human IgG,
IgA, IgM, IgD, or IgE antibody.
[0901] In some cases, two or more AA targeting agents may be linked
to a single full length bivalent Antibody. This is shown below as
Formula III:
Antibody[-Linker'-[AA targeting agent]].sub.2 (III)
[0902] Also provided are stereoisomers, tautomers, solvates,
prodrugs, and pharmaceutically acceptable salts thereof.
[0903] In compounds of Formula III, [AA targeting agent], Linker'
and Antibody are each defined as in Formula II.
[0904] Targeting compounds such as those of Formula II may also be
readily synthesized by covalently linking a targeting agent-linker
compound as described herein to a combining site of a multivalent
antibody. For example, an AA targeting-agent linker conjugate,
where the linker includes a diketone reactive moiety, can be
incubated with 0.5 equivalents of an aldolase antibody, such as
h38C2 IgG1 to produce an AA targeting compound. Alternatively, an
AA targeting compound such as those of Formula III may be produced
by covalently linking an AA targeting agent-linker compound as
described herein to each combining site of a bivalent antibody.
Methods of Use for AA Targeting Compounds
[0905] One aspect of the invention provides methods for modulating
Ang-2 activity in vivo comprising administering an effective amount
of an AA targeting compound as described herein to a subject. There
are further provided methods for treating abnormal angiogenesis or
an angiogenesis-mediated condition in a subject. Such methods
include administering to the subject a therapeutically effective
amount of an AA targeting compound as described herein. As used
herein, an angiogenesis-mediated condition is a condition that is
caused by abnormal angiogenesis activity or one in which compounds
that modulate angiogenesis activity have therapeutic use. Diseases
and conditions that may be treated include cancer, arthritis,
psoriasis, angiogenesis of the eye associated with infection or
surgical intervention, macular degeneration or diabetic
retinopathy. In particular, methods of treating cancer include
carcinomas of the breast, colon, rectum, lung, oropharynx,
hypopharynx, esophagus, stomach, pancreas, liver, gallbladder and
bile ducts, small intestine, urinary tract, female genital tract,
male, genital tract, endocrine glands, and skin; hemangiomas;
melanomas; sarcomas; tumors of the brain, nerves, eyes, and
meninges; leukemia; or lymphoma.
Pharmaceutical Compositions and Methods of Administration
[0906] Another aspect of the invention provides pharmaceutical
compositions of the AA targeting compounds. The AA targeting
compounds can be mixed with pharmaceutically-acceptable carriers to
form a pharmaceutical composition for administration to a cell or
subject, either alone, or in combination with one or more other
modalities of therapy.
[0907] A pharmaceutical composition is generally formulated to be
compatible with its intended route of administration. Those skilled
in the art will know that the choice of the pharmaceutical medium
and the appropriate preparation of the composition will depend on
the intended use and mode of administration. Examples of routes of
administration include parenteral (e.g. intravenous, intramuscular,
intramedullary, intradermal, subcutaneous), oral (e.g. inhalation,
ingestion), intranasal, transdermal (e.g. topical), transmucosal,
and rectal administration. Administration routes of AA targeting
compounds may also include intrathecal, direct intraventricular and
intraperitoneal delivery. The AA targeting compounds may be
administered through any of the parenteral routes either by direct
injection of the formulation or by infusion of a mixture of the
targeting AA compound formulation with an infusion matrix such as
normal saline, D5W, lactated Ringers solution or other commonly
used infusion media.
[0908] The AA targeting compounds may be administered using
techniques well known to those in the art. Preferably, agents are
formulated and administered systemically. Techniques for
formulation and administration may be found in "Remington's
Pharmaceutical Sciences," 18.sup.th Ed., 1990, Mack Publishing Co.,
Easton, Pa. For injection, AA targeting compounds may be formulated
in aqueous solutions, emulsions or suspensions. AA targeting
compounds are preferably formulated in aqueous solutions containing
physiologically compatible buffers such as citrate, acetate,
histidine or phosphate. Where necessary, such formulations may also
contain various tonicity adjusting agents, solubilizing agents
and/or stabilizing agents (e.g. salts such as sodium chloride or
sugars such as sucrose, mannitol, and trehalose, or proteins such
as albumin or amino acids such as glycine and histidine or
surfactants such as polysorbates (Tweens) or cosolvents such as
ethanol, polyethylene glycol and propylene glycol.
[0909] The pharmaceutical composition may contain formulation
materials for modifying, maintaining or preserving, for example,
the pH, osmolarity, viscosity, clarity, color, isotonicity, odor,
sterility, stability, rate of dissolution or release, adsorption or
penetration of the composition. Suitable formulation materials
include, but are not limited to, amino acids (such as glycine,
glutamine, asparagine, arginine or lysine); antimicrobials;
antioxidants (such as ascorbic acid, sodium sulfite or sodium
hydrogen-sulfite); buffers (such as borate, bicarbonate, Tris-HCl,
citrates, phosphates, other organic acids, chelating agents [such
as ethylenediamine tetraacetic acid (EDTA)]; solvents (such as
glycerin, propylene glycol or polyethylene glycol); sugar alcohols
(such as mannitol or sorbitol); suspending agents; surfactants or
wetting agents (such as pluronics, PEG, sorbitan esters,
polysorbates such as polysorbate 20, polysorbate 80, triton,
tromethamine, lecithin, cholesterol, tyloxapal); stability
enhancing agents (sucrose or sorbitol); tonicity enhancing agents
(such as alkali metal halides (preferably sodium or potassium
chloride, mannitol sorbitol); delivery vehicles; diluents;
excipients and/or pharmaceutical adjuvants. (Remington's
Pharmaceutical Sciences, 18th Edition, A. R. Gennaro, ed., Mack
Publishing Company, 1990).
[0910] When parenteral administration is contemplated, the
therapeutic compositions may be in the form of a pyrogen-free,
parenterally acceptable aqueous solution comprising an AA targeting
compound in a pharmaceutically acceptable vehicle. One vehicle for
parenteral injection is sterile distilled water in which an AA
targeting compound is formulated as a sterile, isotonic solution.
Yet another formulation can involve the formulation an AA targeting
compound with an agent, such as injectable microspheres,
bio-degradable particles, polymeric compounds (polylactic acid,
polyglycolic acid), beads, or liposomes, that provides for the
controlled or sustained release of the product which may then be
delivered via a depot injection. Hyaluronic acid may also be used,
and this may have the effect of promoting sustained duration in the
circulation. Other suitable means for the introduction of the
desired molecule include implantable drug delivery devices.
[0911] In another aspect, pharmaceutical formulations suitable for
parenteral administration may be formulated in aqueous solutions,
preferably in physiologically compatible buffers such as Hanks'
solution, Ringer's solution, or a physiologically buffered saline.
Aqueous injection suspensions may contain substances that increase
the viscosity of the suspension, such as sodium carboxymethyl
cellulose, sorbitol, or dextran. Suitable lipophilic solvents or
vehicles include fatty oils, such as sesame oil, or synthetic fatty
acid esters, such as ethyl oleate, triglycerides, or liposomes.
Non-lipid polycationic amino polymers may also be used for
delivery. Optionally, the suspension may also contain suitable
stabilizers or agents to increase the solubility of the compounds
and allow for the preparation of highly concentrated solutions.
[0912] The pharmaceutical composition to be used for in vivo
administration typically must be sterile. This may be accomplished
by filtration through sterile filtration membranes. Where the
composition is lyophilized, sterilization using this method may be
conducted either prior to or following lyophilization and
reconstitution. The composition for parenteral administration may
be stored in lyophilized form or in solution. In addition,
parenteral compositions generally are placed into a container
having a sterile access port, for example, an intravenous solution
bag or vial having a stopper pierceable by a hypodermic injection
needle.
[0913] Once the pharmaceutical composition has been formulated, it
may be stored in sterile vials as a solution, suspension, gel,
emulsion, solid, or a dehydrated or lyophilized powder. Such
formulations may be stored either in a ready-to-use form or in a
form (e.g., lyophilized) requiring reconstitution prior to
administration.
[0914] One embodiment is directed to kits for producing a
single-dose administration unit. The kits may each contain both a
first container having an AA targeting compound and a second
container having an aqueous formulation. Also included within the
scope of this invention are kits containing single and
multi-chambered pre-filled syringes.
[0915] In treating mammals, including humans, having a disorder
with an angiogenic component to the disorder, a therapeutically
effective amount of an AA targeting compound or a pharmaceutically
acceptable derivative is administered. The frequency of dosing will
depend upon the pharmacokinetic parameters of the AA targeting
compound in the formulation used. Typically, a composition is
administered until a dosage is reached that achieves the desired
effect. The composition may therefore be administered as a single
dose, or as multiple doses (at the same or different
concentrations/dosages) over time, or as a continuous infusion.
Routes and frequency of administration of a composition as well as
dosage may vary from individual to individual and may be readily
established using standard techniques. Further refinement of the
appropriate dosage is routinely made. Appropriate dosages may be
developed by one skilled in the art through the use of appropriate
dose-response data.
[0916] An appropriate dosage and treatment regimen provides the
active compound(s) in an amount sufficient to provide therapeutic
and/or prophylactic benefit. Such a response can be monitored by
establishing an improved clinical outcome (e.g. reduced number of
blood vessels in a target area, decreased tumor size or volume) in
treated patients as compared to non-treated patients. Typically, a
suitable dose is an amount of a compound that, when administered as
described herein, is capable of promoting an anti-angiogenesis
response, and/or is at least 10-50% above the basal or untreated
level.
[0917] In some embodiments, the most effective mode of
administration and dosage regimen for the invention compositions
depends upon the severity and course of the disease, the patient's
health and response to treatment, and the judgment of the treating
physician. Accordingly, the dosages of the invention compositions
should be titrated to the individual patient. An effective dose of
the compounds is in the range of from about 0.1 ug to about 40 mg
per kilogram per day. An AA targeting compound may be administered
as a daily intravenous infusion from about 0.1 mg/kg body weight to
about 15 mg/kg body weight. Accordingly, one embodiment provides a
dose of about 0.5 mg/kg body weight. Another embodiment provides a
dose of about 0.75 mg/kg body weight. Another embodiment provides a
dose of about 1.0 mg/kg body weight. Another embodiment provides a
dose of about 2.5 mg/kg body weight. Another embodiment provides a
dose of about 5 mg/kg body weight. Another embodiment provides a
dose of about 10.0 mg/kg body weight. Another embodiment provides a
dose of about 15.0 mg/kg body weight. Doses of an AA targeting
compound or a pharmaceutically acceptable derivative should be
administered in intervals of from about once per day to 2 times per
week, or alternatively, from about once every week to once per
month. In one embodiment, a dose is administered to achieve peak
plasma concentrations of an AA targeting compound or a
pharmaceutically acceptable derivative thereof from about 0.002
mg/ml to 30 mg/ml. This may be achieved by the sterile injection of
a solution of the administered ingredients in an appropriate
formulation (any suitable formulation solutions known to those
skilled in the art of chemistry may be used). Desirable blood
levels may be maintained by a continuous infusion of an AA
targeting compound as ascertained by plasma levels measured by a
validated analytical methodology.
[0918] One method for administering an AA targeting compound to an
individual comprises administering an AA targeting agent-linker
conjugate to the individual and allowing it to form a covalent
compound with a combining site of an appropriate antibody in vivo.
The antibody portion of an AA targeting compound that forms in vivo
may be administered to the individual before, at the same time, or
after administration of the targeting agent-linker conjugate. As
already discussed, an AA targeting agent may include a
linker/reactive moiety, or the antibody combining site may be
suitably modified to covalently link to the targeting agent.
Alternatively, or in addition, an antibody may be present in the
circulation of the individual following immunization with an
appropriate immunogen. For example, catalytic antibodies may be
generated by immunizing with a reactive intermediate of the
substrate conjugated to a carrier protein. See R. A. Lerner and C.
F. Barbas 3.sup.rd, Acta Chem. Scand. 50:672-678 (1996). In
particular, aldolase catalytic antibodies may be generated by
administering with keyhole limpet hemocyanin linked to a diketone
moiety as described by P. Wirsching et al., Science 270:1775-1782
(1995) (commenting on J. Wagner et al., Science 270:1797-1800
(1995)).
[0919] The invention also provides a method of visualizing or
localizing Ang-2 or anti-angiogenesis target (i.e. AA-targeting
agent receptor) in tissues and cells. In one embodiment, biopsied
tissues may be examined for presence of AA-targeting agent
receptor. In another embodiment, neovascularization in a subject
may be imaged by administering to the subject an AA targeting agent
or compound including a detectable label. As used herein, the term
"detectable label" refers to any molecule which can be administered
in vivo and subsequently detected. Exemplary detectable labels
include radiolabels and fluorescent molecules. Exemplary
radionuclides include indium-111, technetium-99, carbon-11, and
carbon-13. Fluorescent molecules include, without limitation,
fluorescein, allophycocyanin, phycoerythrin, rhodamine, and Texas
red.
Combination Therapies
[0920] The vasculature within a tumor generally undergoes active
angiogenesis, resulting in the continual formation of new blood
vessels to support the growing tumor. Such angiogenic blood vessels
are distinguishable from mature vasculature in that angiogenic
vasculature expresses unique endothelial cell surface markers,
including the .A-inverted..sub.v.E-backward..sub.3 integrin.
(Brooks, Cell 79:1157-1164 (1994); WO 95/14714, Int. Filing Date
Nov. 22, 1994) and receptors for angiogenic growth factors
(Mustonen and Alitalo, J. Cell Biol. 129:895-898 (1995); Lappi,
Semin. Cancer Biol. 6:279-288 (1995)).
[0921] The invention also includes administration of one or more AA
targeting agents in combination with one or more oncology
therapeutics, each being administered according to a regimen
suitable for that therapeutic. The components of the combination
therapy may be administered concurrently or non-concurrently. As
used herein, the terms "concurrently administered" and "concurrent
administration" encompass substantially simultaneous administration
of one or more AA targeting compounds and one other oncology
therapeutic.
[0922] As used herein, the term, "non-concurrent" administration
encompasses administering one or more AA targeting compounds at
different times, in any order, whether overlapping or not. This
includes, but is not limited to, sequential treatment (such as
pretreatment, post-treatment, or overlapping treatment) with the
components of the combination, as well as regimens in which the
drugs are alternated, or wherein one component is administered
long-term and the other(s) are administered intermittently.
Components may be administered in the same or in separate
compositions, and by the same or different routes of
administration.
[0923] Suitable oncology therapeutics and combinations that may be
used in combination with an AA targeting compound are listed in
Tables 4-6.
TABLE-US-00023 TABLE 4 Approved oncology drugs and indications
Generic Trade Name Indication Company Aldesleukin Proleukin
Proleukin is indicated for the Chiron Corp treatment of adults with
metastatic renal cell carcinoma (metastatic RCC) and for the
treatment of adults with metastatic melanoma. Alemtuzumab Campath
Campath is indicated for the Millennium and treatment of B-cell
chronic ILEX Partners, LP lymphocytic leukemia (B-CLL) in patients
who have been treated with alkylating agents and who have failed
fludarabine therapy. Alitretinoin Panretin Topical treatment of
cutaneous Ligand lesions in patients with AIDS- Pharmaceuticals
related Kaposi's sarcoma. Allopurinol Zyloprim Patients with
leukemia, GlaxoSmithKline lymphoma and solid tumor malignancies who
are receiving cancer therapy which causes elevations of serum and
urinary uric acid levels and who cannot tolerate oral therapy.
Palonosetron Aloxi For the treatment of nausea MGI Pharmaceuticals
Altretamine Hexalen Single agent palliative US Bioscience treatment
of patients with persistent or recurrent ovarian cancer following
first-line therapy with a cisplatin and/or alkylating agent based
combination. Amifostine Ethyol To reduce the cumulative renal US
Bioscience toxicity associated with repeated administration of
cisplatin in patients with advanced ovarian cancer Amifostine
Ethyol Reduces platinum toxicity in US Bioscience non-small cell
lung cancer Amifostine Ethyol To reduce post-radiation US
Bioscience xerostomia for head and neck cancer where the radiation
port includes a substantial portion of the parotid glands.
Anastrozole Arimidex Adjuvant treatment of AstraZeneca
postmenopausal women with hormone receptor positive early breast
cancer Anastrozole Arimidex Treatment of advanced breast
AstraZeneca cancer in postmenopausal Pharmaceuticals women with
disease progression following tamoxifen therapy. Anastrozole
Arimidex For first-line treatment of AstraZeneca postmenopausal
women with Pharmaceuticals hormone receptor positive or hormone
receptor unknown locally advanced or metastatic breast cancer.
Nelarabine Arranon For the treatement of T cell GlaxoSmithKline
acute lymphoblatic leukemia Arsenic trioxide Trisenox Second line
treatment of Cell Therapeutic relapsed or refractory APL following
ATRA plus an anthracycline. Asparaginase Elspar ELSPAR is indicated
in the Merck & Co, Inc therapy of patients with acute
lymphocytic leukemia. This agent is useful primarily in combination
with other chemotherapeutic agents in the induction of remissions
of the disease in pediatric patients. Bevacizumab Avastin For the
treatment of metastatic Genentech colorectal cancer Bexarotene
Targretin For the treatment by oral Ligand capsules capsule of
cutaneous Pharmaceuticals manifestations of cutaneous T- cell
lymphoma in patients who are refractory to at least one prior
systemic therapy. Bexarotene gel Targretin For the topical
treatment of Ligand cutaneous manifestations of Pharmaceuticals
cutaneous T-cell lymphoma in patients who are refractory to at
least one prior systemic therapy. Bleomycin Blenoxane Palliative
agent for the Bristol-Myers management of the following Squibb
neoplasms: Squamous Cell Carcinoma (head and neck including mouth,
tongue, tonsil, nasopharynx, oropharynx, sinus, palate, lip, buccal
mucosa, gingivae, epiglottis, skin, larynx, penis, cervix, and
vulva. Lymphomas (Hodgkin's Disease, non-Hodgkin's lymphoma).
Testicular Carcinoma (Embryonal cell, choriocarcinoma, and
teratocarcinoma). Bleomycin Blenoxane Sclerosing agent for the
Bristol-Myers treatment of malignant pleural Squibb effusion (MPE)
and prevention of recurrent pleural effusions. Busulfan Busulfex
Use in combination with Orphan Medical, intravenous cyclophoshamide
as Inc. conditioning regimen prior to allogeneic hematopoietic
progenitor cell transplantation for chronic myelogenous leukemia.
Busulfan oral Myleran Palliative therapy for Chronic
GlaxoSmithKline Myelogenous Leukemia - Calusterone Methosarb
Synthetic androgen for the Pharmacia & treatment of androgen
sensitive Upjohn Company cancers Capecitabine Xeloda Treatment of
metastatic breast Roche cancer resistant to both paclitaxel and an
anthracycline containing chemotherapy regimen or resistant to
paclitaxel and for whom further anthracycline therapy may be
contraindicated, e.g., patients who have received cumulative doses
of 400 mg/m2 of doxorubicin or doxorubicin equivalents Capecitabine
Xeloda Initial therapy of patients with Roche metastatic colorectal
carcinoma when treatment with fluoropyrimidine therapy alone is
preferred. Combination chemotherapy has shown a survival benefit
compared to 5- FU/LV alone. A survival benefit over 5_FU/LV has not
been demonstrated with Xeloda monotherapy. Capecitabine Xeloda
Treatment in combination with Roche docetaxel of patients with
metastatic breast cancer after failure of prior anthracycline
containing chemotherapy Carboplatin Paraplatin Palliative treatment
of patients Bristol-Myers with ovarian carcinoma Squibb recurrent
after prior chemotherapy, including patients who have been
previously treated with cisplatin. Carboplatin Paraplatin Initial
chemotherapy of Bristol-Myers advanced ovarian carcinoma in Squibb
combination with other approved chemotherapeutic agents. Carmustine
BCNU, BiCNU Palliative therapy as a single Bristol-Myers agent or
in established Squibb combination therapy with other approved
chemotherapeutic agents in the following: Brain tumors
(glioblastoma, brainstem glioma, medulloblastoma, astrocytoma,
ependymoma, and metastatic brain tumors); Multiple myeloma;
Hodgkin's Disease; and Non-Hodgkin's lymphomas. Carmustine with
Giladel Wafer For use in addition to surgery Guilford Polifeprosan
20 to prolong survival in patients Pharmaceuticals Implant with
recurrent glioblastoma Inc. multiforme who qualify for surgery.
Celecoxib Celebrex Reduction of polyp number in Searle patients
with the rare genetic disorder of familial adenomatous polyposis.
Cetuximab Erbitux For the treatement of EGFR expressing metastatic
colorectal cancer Chlorambucil Leukeran Chronic Lymphocytic
GlaxoSmithKline Leukemia-palliative therapy Chlorambucil Leukeran
Treatment for CLL or indolent GlaxoSmithKline NHL. Cinacalchet
Sensipar For the treatment of secondary Amgen hypparathyroidism
Cisplatin Platinol Metastatic testicular-in Bristol-Myers
established combination Squibb therapy with other approved
chemotherapeutic agents in patients with metastatic testicular
tumors whoc have already received appropriate surgical and/or
radiotherapeutic procedures. An established combination therapy
consists of Platinol, Blenoxane and Velbam. Cisplatin Platinol
Metastatic ovarian tumors - in Bristol-Myers established
combination Squibb therapy with other approved chemotherapeutic
agents: Ovarian-in established combination therapy with other
approved chemotherapeutic agents in patients with metastatic
ovarian tumors who have already received appropriate surgical
and/or radiotherapeutic procedures. An established combination
consists of Platinol and Adriamycin. Platinol, as a single agent,
is indicated as secondary therapy in patients with metastatic
ovarian tumors refractory to standard chemotherapy who have not
previously received Platinol therapy. Cisplatin Platinol
Transitional cell bladder cancer Bristol-Myers which is no longer
amenable to Squibb local treatments such as surgery and/or
radiotherapy. Cladribine Leustatin, 2-CdA Treatment of active hairy
cell R. W. Johnson leukemia. Pharmaceutical Research Institute
Clofarabine Clolar Treatment for acute Genzyme lymphblastic
leukemia Cyclophosphamide Cytoxan, Neosar Treatment for ovary,
breast, Bristol-Myers bladder and CLL. Squibb Cytarabine Cytosar-U
Treatment for AML Pharmacia & Upjohn Company Cytarabine DepoCyt
Intrathecal therapy of Skye Liposomal lymphomatous meningitis
Pharmaceuticals Dacarbazine DTIC-Dome Treatment for melanoma and
Bayer Hodgkins lymphoma Dactinomycin, Cosmegan Treatment for
pediatric Merck
actinomycin D leukemias Darbepoetin alfa Aranesp Treatment of
anemia Amgen, Inc. associated with chronic renal failure.
Darbepoetin alfa Aranesp Aranesp is indicated for the Amgen, Inc.
treatment of anemia in patients with non-myeloid malignancies where
anemia is due to the effect of concomitantly administered
chemotherapy. Daunorubicin DanuoXome First line cytotoxic therapy
for Nexstar, Inc. liposomal advanced, HIV related Kaposi's sarcoma.
Daunorubicin, Daunorubicin Leukemia/myelogenous/monocytic/ Bedford
Labs' daunomycin erythroid of adults/remission induction in acute
lymphocytic leukemia of children and adults. Daunorubicin,
Cerubidine In combination with approved Wyeth Ayerst daunomycin
anticancer drugs for induction of remission in adult ALL.
Danileukin Ontak Treatment of patients with Seragen, Inc. diftitox
persistent or recurrent cutaneous T-cell lymphoma whose malignant
cells express the CD25 component of the IL- 2 receptor Dexrazoxane
Zinecard Prevention of cardiomyopathy Pharmacia & associated
with doxorubicin Upjohn Company administration Dexrazoxane Zinecard
Used for reducing the Pharmacia & incidence and severity of
Upjohn Company cardiomyopathy associated with doxorubicin
administration in women with metastatic breast cancer who have
received a cumulative doxorubicin dose of 300 mg/m2 and who will
continue to receive doxorubicin therapy to maintain tumor control.
Docetaxel Taxotere Treatment of patients with Aventis locally
advanced or metastatic Pharmaceutical breast cancer who have
progressed during anthracycline-based therapy or have relapsed
during anthracycline-based adjuvant therapy. Docetaxel Taxotere For
the treatment of locally Aventis advanced or metastatic breast
Pharmaceutical cancer which has progressed during
anthracycline-based treatment or relapsed during
anthracycline-based adjuvant therapy. Docetaxel Taxotera For
locally advanced or Aventis metastatic non-small cell lung
Pharmaceutical cancer after failure of prior platinum-based
chemotherapy. Docetaxel Taxotere Aventis Pharmaceutical Docetaxel
Taxotere Used in combination with Aventis cisplatin for the
treatment of Pharmaceutical patients with unresectable, locally
advanced or metastatic non-small cell lung cancer who have not
previously received chemotherapy for this condition. Doxorubicin
Adriamycin PFS Antibiotic, antitumor agent. Pharmacia &
Injection Upjohn Company intravenous injection Doxorubicin Doxil
Treatment of AIDS-related Sequus liposomal Kaposi's sarcoma in
patients Pharmaceuticals, with disease that has Inc. progressed on
prior combination chemotherapy or in patients who are intolerant to
such therapy. Doxorubicin Doxil Treatment of metastatic Sequus
liposomal carcinoma of the ovary in Pharmaceuticals, patient with
disease that is Inc. refractory to both paclitaxel and platinum
based regimens Dromostanolone Dromostanolone Sythetic androgen for
use in Eli Lilly Propionate androgen sensitve cancers Elliott's B
Elliott's B Diluent for the intrathecal Orphan Medical, Solution
Solution administration of methotrexate Inc. sodium and cytarabine
for the prevention or treatment of meningeal leukemia or
lymphocytic lymphoma. Epoetin Epogen EPOGEN is indicated for the
Amgen, Inc. alfa/beta treatment of anemia. Erlotinib Tarceva For
the treatment of advanced OSI metatstaic non-small cell lung
Pharmaceuticals cancer Estramustine Emcyt Palliation of prostate
cancer Pharmacia & Upjohn Company Etoposide Etopophos
Management of refractory Bristol-Myers phosphate testicular tumors,
in Squibb combination with other approved chemotherapeutic agents.
Etoposide Etopophos Management of small cell lung Bristol-Myers
phosphate cancer, first-line, in Squibb combination with other
approved chemotherapeutic agents. Etoposide Etopophos Management of
refractory Bristol-Myers phosphate testicular tumors and small cell
Squibb lung cancer. Etoposide, VP- Vepesid Refractory testicular
tumors-in Bristol-Myers 16 combination therapy with other Squibb
approved chemotherapeutic agents in patients with refractory
testicular tumors who have already received appropriate surgical,
chemotherapeutic and radiotherapeutic therapy. Etoposide, VP-
VePesid In combination with other Bristol-Myers 16 approved
chemotherapeutic Squibb agents as first line treatment in patients
with small cell lung cancer. Etoposide, VP- Vepesid In combination
with other Bristol-Myers 16 approved chemotherapeutic Squibb agents
as first line treatment in patients with small cell lung cancer.
Exemestane Aromasin Treatment of advance breast Pharmacia &
cancer in postmenopausal Upjohn Company women whose disease has
progressed following tamoxifen therapy. Filgrastim Neupogen
NEUPOGEN is indicated for Amgen, Inc. reducing the time to
neutrophil recovery and the duration of fever, following induction
or consolidation hemotherapy treatment of adults with AML.
Floxuridine FUDR An analog for 5-flurouracil. Roche (intraarterial)
FUDR has been approved in the directed treatment of liver
metastases using hepatic arterial infusion. Fludarabine Fludara
Palliative treatment of patients Berlex with B-cell lymphocytic
Laboratories Inc. leukemia (CLL) who have not responded or have
progressed during treatment with at least one standard alkylating
agent containing regimen. Fluorouracil, 5- Adrucil Prolong survival
in combination ICN Puerto Rico FU with leucovorin Fulvestrant
Faslodex the treatment of hormone IPR receptor-positive metastatic
breast cancer in postmenopausal women with disease progression
following antiestrogen therapy Gemcitabine Gemzar Treatment of
patients with Eli Lilly locally advanced (nonresectable stage II or
III) or metastatic (stage IV) adenocarcinoma of the pancreas.
Indicated for first-line treatment and for patients previously
treated with a 5- fluorouracil-containing regimen. Gemcitabine
Gemzar For use in combination with Eli Lilly cisplatin for the
first-line treatment of patients with inoperable, locally advanced
(Stage IIIA or IIIB) or metastatic (Stage IV) non-small cell lung
cancer. Gemtuzumab Mylotarg Treatment of CD33 positive Wyeth Ayerst
ozogamicin acute myeloid leukemia in patients in first relapse who
are 60 years of age or older and who are not considered candidates
for cytotoxic chemotherapy. Goserelin Zoladex implant Palliative
treatment of AstraZeneca acetate advanced breast cancer in pre-
Pharmaceuticals and perimenopausal women. Goserelin Zoladex Used
for treatement of prostate AstraZeneca acetate cancer
Pharmaceuticals Hydroxyurea Hydrea Decrease need for transfusions
Bristol-Myers in sickle cell anemia Squibb Ibritumomab Zevalin
Treatment of patients with IDEC tiuxetan relapsed or refractory
low- Pharmaceuticals grade, follicular, or transformed Corp. B-cell
non-Hodgkin's lymphoma, including patients with Rituximab
refractory follicular non-Hodgkin's lymphoma. Idarubicin Idamycin
For use in combination with Adria Laboratories other approved
antileukemic drugs for the treatment of acute myeloid leukemia
(AML) in adults. Idarubicin Idamycin In combination with other
Pharmacia & approved antileukemic drugs Upjohn Company for the
treatment of acute non- lymphocytic leukemia in adults. Ifosfamide
IFEX Third line chemotherapy of Bristol-Myers germ cell testicular
cancer Squibb when used in combination with certain other approved
antineoplastic agents. Imatinib Gleevec Initial therapy of chronic
Novartis mesylate myelogenous leukemia Imatrinib Gleevac Treatment
of metastatic or Novartis mesylate unresectable malignant
gastrointestinal stromal tumors Imatinib Gleevec Initial treatment
of newly Novartis mesylate diagnosed Ph+ chronic myelogenous
leukemia (CML). Interferon alfa- Roferon-A Treatment of chronic
Hoffmann-La 2a hepatitis C, hairy cell leukemia Roche Inc. and
AIDS-related Kaposi's sarcoma in adult patients and for chronic
phase, Philadelphia chromosome (Ph) positive chronic myelogenous
leukemia (CML) patients who are minimally pretreated (within 1 year
of diagnosis). Interferon alfa- Intron A Interferon alfa-2b,
recombinant Schering Corp. 2b for injection is indicated as
adjuvant to surgical treatment in patients 18 years of age or older
with malignant melanoma who are free of disease but at high risk
for systemic recurrence within 56 days of surgery. Interferon
alfa-2b, recombinant for Injection is indicated for the initial
treatment of clinically aggressive follicular Non- Hodgkin's
Lymphoma in conjunction with anthracycline- containing combination
chemotherapy in patients 18
years of age or older. Interferon alfa-2b, recombinant for
Injection is indicated for intralesional treatment of selected
patients 18 years of age or older with condylomata acuminata
involving external surfaces of the genital and perianal areas.
Interferon alfa-2b, recombinant for Injection is indicated for the
treatment of patients 18 years of age or older with hairy cell
leukemia. Interferon alfa-2b, recombinant for Injection is
indicated for the treatment of selected patients 18 years of age or
older with AIDS-Related Kaposi's Sarcoma. The likelihood of
response to INTRON A therapy is greater in patients who are without
systemic symptoms, who have limited lymphadenopathy and who have a
relatively intact immune system as indicated by total CD4 count.
Irinotecan Camptosar Treatment of patients with Pharmacia &
metastatic carcinoma of the Upjohn Company colon or rectum whose
disease has recurred or progressed following 5-FU-based therapy.
Letrozole Femara First-line treatment of Novartis postmenopausal
women with hormone receptor positive or hormone receptor unknown
locally advanced or metastatic breast cancer. Letrozole Femara Used
for treatment of post- Novartis menopausal women with early stage
breast cancer Leucovorin Wellcovorin, Leucovorin calcium is
indicated Immunex Leucovorin fro use in combination with 5-
Corporation fluorouracil to prolong survival in the palliative
treatment of patients with advanced colorectal cancer. Leucovorin
Leucovorin In combination with fluorouracil Lederle to prolong
survival in the laboratories palliative treatment of patients with
advanced colorectal cancer. Levamisole Ergamisol Adjuvant treatment
in Janssen combination with 5-fluorouracil Research after surgical
resection in Foundation patients with Dukes' Stage C colon cancer.
Lomustine, CeeNu An alkylating agent used for the Bristol-Myers
CCNU treatment of brain cancer and Squibb NHL. Meclorethamine,
Mustargen A nitrogen mustard used in the Merck nitrogen treatment
of lymphoma. mustard Megestrol Megace A synthetic progesterone used
Bristol-Myers acetate for the treatment of estrogen Squibb
sensitive cancers. Melphalan, L- Alkeran Systemic administration
for GlaxoSmithKline PAM palliative treatment of patients with
multiple myeloma for whom oral therapy is not appropriate.
Mercaptopurine, Purinethol Purinethol is indicated for
GlaxoSmithKline 6-MP remission induction and maintenance therapy of
acute lymphatic leukemia. Mesna Mesnex Prevention of ifosfamide-
Asta Medica induced hemorrhagic cystitis Methotrexate Methotrexate
Is used to treat cancer of the Laderle breast, head and neck, lung,
Laboratories blood, bone, and lymph, and tumors in the uterus.
Methoxsalen Uvadex For the use of UVADEX with Therakos the UVAR
Photopheresis System in the palliative treatment of the skin
manifestations of cutaneous T- cell lymphoma (CTCL) that is
unresponsive to other forms of treatment. Mitromycin C Mitozytrex
Therapy of disseminated Supergen adenocarcinoma of the stomach or
pancreas in proven combinations with other approved
chemotherapeutic agents and as palliative treatment when other
modalities have failed. Mitotane Lysodren Used for the treatment of
Bristol-Myers adrenal cancers. Squibb Mitoxantrone Novantrone For
use in combination with Immunex corticosteroids as initial
Corporation chemotherapy for the treatment of patients with pain
related to advanced hormone-refractory prostate cancer.
Mitoxantrone Novantrone For use with other approved Laderle drugs
in the initial therapy for Laboratories acute nonlymphocytic
leukemia (ANLL) in adults. Nandrolone Durabolin-509 It is indicated
as a treatment for Organon phenpropionate palliation of inoperable
metastatic breast cancer in postmenopausal women. Nofetumomab
Verluma Verluma is a monoclonal Boehringer antibody Fab fragment
linked to Ingelheim Pharma .sup.99mTc. Verluma identifies KG
(formerly Dr. advanced-stage disease in Karl Thomae patients with
small-cell lung GmbH) cancer (SCLC). Oprelvekin Neumega Neumega is
indicated for the Genetics Institute, prevention of severe Inc.
thrombocytopenia and the reduction of the need for platelet
transfusions following myelosuppressive chemotherapy in adult
patients with nonmyeloid malignancies who are at high risk of
severe thrombocytopenia. Oxaliplatin Eloxatin Used) in combination
with Sanofi Synthelabo infusional 5-FU/LV, is indicated for the
treatment of patients with metastatic carcinoma of the colon or
rectum whose disease has recurred or progressed during or within 6
months of completion of first line therapy with the combination of
bolus 5-FU/LV and irinotecan. Paclitaxel Paxene Treatment of
advanced AIDS- Baker Norton related Kaposi's sarcoma after
Pharmaceuticals, failure of first line or Inc. subsequent systemic
chemotherapy Paclitaxel Taxol Treatment of patients with
Bristol-Myers metastatic carcinoma of the Squibb ovary after
failure of first-line or subsequent chemotherapy. Treatment of
breast cancer after failure of combination chemotherapy for
metastatic disease or relapse within 6 months of adjuvant
chemotherapy. Prior therapy should have included an anthracycline
unless clinically contraindicated. New dosing regimen for patients
who have failed initial or subsequent chemotherapy for metastatic
carcinoma of the ovary Second line therapy for AIDS related
Kaposi's sarcoma. For first-line therapy for the treatment of
advanced carcinoma of the ovary in combination with cisplatin. For
use in combination with cisplatin, for the first-line treatment of
non-small cell lung cancer in patients who are not candidates for
potentially curative surgery and/or radiation therapy. For the
adjuvant treatment of node-positive breast cancer administered
sequentially to standard doxorubicin- containing combination
therapy. First line ovarian cancer with 3 hour infusion.
Pamidronate Aredia Treatment of osteolytic bone Novartis metastases
of breast cancer in conjunction with standard antineoplastic
therapy. Pegademase Adagen Enzyme replacement therapy Enzon
(Pegademase for patients with severe Bovine) combined
immunodeficiency asa result of adenosine deaminase deficiency.
Pegaspargase Oncaspar PEG asparginase used in the Enzon, Inc.
treatment of ALL. Pegfilgrastim Neulasta Neulasta is indicated to
Amgen, Inc. decrease the incidence of infection, as manifested by
febrile neutropenia, in patients with non-myeloid malignancies
receiving myelosuppressive anti-cancer drugs associated with a
clinically significant incidence of febrile neutropenia. Pemetrexed
Alimta Treatment of malignant pleural Eli Lilly mesothelioma
Pentostatin Nipent Single agent treatment for adult Parke-Davis
patients with alpha interferon Pharmaceutical refractory hairy cell
leukemia. Co. Pipobroman Vercyte Used in the treatment of CRC.
Abbott Labs Plicamycin, Mithracin Used in the treatment of Pfizer
Labs mithramycin testicular cancer. Porfimer sodium Photofrin For
use in photodynamic QLT therapy (PDT) for palliation of
Phototherapeutics patients with completely Inc. obstructing
esophageal cancer, or patients with partially obstructing
esophageal cancer who cannot be satisfactorily treated with ND-YAG
laser therapy. For use in photodynamic therapy for treatment of
microinvasive endobronchial nonsmall cell lung cancer in patients
for whom surgery and radiotherapy are not indicated. For use in
photodynamic therapy (PDT) for reduction of obstruction and
palliation of symptoms in patients with completely or partially
obstructing endobroncial nonsmall cell lung cancer (NSCLC).
Procarbazine Matulane One component of the MOPP Sigma Tau regime.
Pharms Rasburicase Elitek ELITEK is indicated for the Sanofi-
initial management of plasma Synthelabo, Inc. uric acid levels in
pediatric patients with leukemia, lymphoma, and solid tumor
malignancies who are receiving anti-cancer therapy expected to
result in tumor lysis and subsequent elevation of plasma uric acid.
Rituximab Rituxan Used in the treatment NHL. Genentech, Inc.
Sargramostim Prokine GM-CSF used in the treatment Immunex Corp.
of NHL, Hodgkins Leukemia and acute lymphoblastic leukemia.
Sorafenib Nexavar Treatment of RCC Bayer/Onyx Streptozocin Zanosar
Antineoplastic agent. Pharmacia & Upjohn Company Talc Slerosol
For the prevention of the Bryan recurrence of malignant pleural
effusion in symptomatic patients. Tamoxifen Nolvadex As a single
agent to delay AstraZeneca breast cancer recurrence Pharmaceuticals
following total mastectomy and axillary dissection in
postmenopausal women with breast cancer (T1-3, N1, M0). For use in
premenopausal women with metastatic breast cancer as an alternative
to oophorectomy or ovarian irradiation. For use in women with
axillary node-negative breast cancer adjuvant therapy. Metastatic
breast cancer in men. Temozolomide Temodar For treatment of adult
patients Scherine with refractory anaplastic astrocytoma, i.e.,
patients at first relapse with disease progression on a nitrosourea
and procarbazine containing regimen Teniposide, VM- Vumon In
combination with other Bristol-Myers 26 approved anticancer agents
for Squibb induction therapy in patients with refractory childhood
acute lymphoblastic leukemia (all). Testolactone Teslac Used in the
treatment of breast Bristol-Myers cancer. Squibb Thioguanine, 6-
Thioguanine Antimetabolite used in the GlaxoSmithKline TG treatment
of AML, CML, CLL. Thiotepa Thioplex Thiotepa is a cytotoxic agent
of Immunex the polyfunctional type, related Corporation chemically
and pharmacologically to nitrogen mustard. Thiotepa has been tried
with varying results in the palliation of a wide variety of
neoplastic diseases. However, the most consistent results have been
seen in the following tumors: 1. Adenocarcinoma of the breast. 2.
Adenocarcinoma of the ovary. 3. For controlling intracavitary
effusions secondary to diffuse or localized neoplastic diseases of
various serosal cavities. 4. For the treatment of superficial
papillary carcinoma of the urinary bladder. While now largely
superseded by other treatments, thiotepa has been effective against
other lymphomas, such as lymphosarcoma and Hodgkin's disease.
Topotecan Hycamtin Treatment of patients with GlaxoSmithKline
metastatic carcinoma of the ovary after failure of initial or
subsequent chemotherapy. Treatment of small cell lung cancer
sensitive disease after failure of first-line chemotherapy.
Toremifene Fareston Treatment of advanced breast Chiron Corp.
cancer in postmenopausal women. Tositumomab Bexxar Accel. Approv.
(clinical benefit Corixa not established) Treatment of Corporation
patients with CD20 positive, follicular, non-Hodgkin's lymphoma,
with and without transformation, whose disease is refractory to
Rituximab and has relapsed following chemotherapy Trastuzumab
Herceptin HERCEPTIN as a single agent Genentech, Inc. is indicated
for the treatment of patients with metastatic breast cancer whose
tumors overexpress the HER2 protein and who have received one or
more chemotherapy regimens for their metastatic disease. Herceptin
in combination with paclitaxel is indicated for treatment of
patients with metastatic breast cancer whose tumors overexpress the
HER-2 protein and had not received chemotherapy for their
metastatic disease Tretinoin, ATRA Vesanoid Induction of remission
in Roche patients with acute promyelocytic leukemia (APL) who are
refractory to or unable to tolerate anthracycline based cytotoxic
chemotherapeutic regimens. Uracil Mustard Uracil Mustard Used in
the treatment of CML, Roberts Labs Capsules NHL and CLL. Valrubicin
Valstar For intravesical therapy of Anthra .fwdarw. Medeva
BCG-refractory carcinoma in situ (CIS) of the urinary bladder in
patients for whom immediate cystectomy would be associated with
unacceptable morbidity or mortality. Vinblastine Velban Vinca
alkyloid used in the Eli Lilly treatment of many types of cancer.
Vincristine Oncovin Vinca alkyloid used in the Eli Lilly treatment
of many types of cancer. Vinorelbine Navelbine Single agent or in
combination GlaxoSmithKline with cisplatin for the first-line
treatment of ambulatory patients with unresectable, advanced
non-small cell lung cancer (NSCLC). Vinorelbine Navelbine Navelbine
is indicated as a GlaxoSmithKline single agent or in combination
with cisplatin for the first-line treatment of ambulatory patients
with unreseactable, advanced non-small cell lung cancer (NSCLC). In
patients with Stage IV NSCLC, Navelbine is indicated as a single
agent or in combination with cisplatin. In Stage III NSCLC,
Navelbine is indicated in combination with cisplatin. Zoledronate
Zometa Used in the treatment of Novartis patients with multiple
myeloma and patients with documented bone metastases from solid
tumors, in conjunction with standard antineoplastic therapy.
Prostate cancer should have progressed after treatment with at
least one hormonal therapy
TABLE-US-00024 TABLE 5 Advanced antiangiogenic compounds in the
clinic Clinical Product Mechanism of Action Phase Marketing Co.
Sorafenib Inhibits VEGFR2, VEGFR3, Pre- Bayer/Onyx Raf Kinase and
PDGFRa registration Sutent Inhibits VEGFR1, VEGFR2, Pre- Pfizer
VEGFR3, PDGFR, CSF-1, Fit- registration 3, and C-Kit Thalomid
Antiangiogenic compound of III Celgene unknown mechanism of action
Revlimid Antiangiogenic compound of III Celgene unknown mechanism
of action Vatalanib Inhibits VEGFR1, VEGFR2, III Novartis/Schering
VEGFR3, PDGFR, and C-Kit ZD-6474 Inhibits VEGFR2, and EGFR III
AstraZeneca Neovastat Liquid extract derived from III AEterna Shark
cartilage that blocks VEGFR2 and inhibits MMP-1, MMP-9 and MMP-12
GSK-786024 Inihibits VEGFR1, VEGFR2 and II GlaxoSmithKline VEGFR3
AEE-788 Inhibits EGFR, HER2 and II Novartis VEGFR AG-13736
Inihibits VEGFR1, VEGFR2 and II Pfizer PDGF AMG706 Inhibits VEGFR1,
VEGFR2, II Amgen VEGFR3, PDGFR, Ret, and C- Kit AZD-2171 Inhibits
VEGFR1, VEGFR2, II AstraZeneca VEGFR3, and EGFR BIBF-1120 Inhibits
VEGFR, FGFR, and II Boehringer PDGFR Ingelheim CP-547,632 Inhibits
VEGFR1 and VEGFR2 II Pfizer/OSI Pharma Midostaurin Inhibits FLT3
Kinase, VEGFR2, II Novartis and various PKC kinases SU-6668
Inhibits VEGFR1, PDGF and II Pfizer/Taiho FGFR CDP-791 Inhibits
VEFR2 II UCB/Imclone Systems PI-88 Inhibits heparinase, binds to II
Progen VEGF, FGF1, FGF2 and stimulates the release of TFP1 PCK-3145
Binds to laminin receptor and II Procyon Biopharma VEGFR2, and
downregulates MMP9 expression Atiprimod Inhibits IL6 and VEGF
secretion II Callisto Pharmaceuticals A6 Eight amino acid, uPA
derived II Angstrom peptide that inhibits the activity
Pharmaceuticals of uPAR Angiostatin Peptidic angiostatin inhibitor
II Alchemgen that is a fragment of the clotting Therapeutics factor
plasminogen Cilengitide Cyclic Peptide that is an alpha-v II Merck
integrin antagonist Enodstatin Peptidic angiogenesis inhibitor II
Alchemgen based Collagen XVIII fragment Therapeutics rPF4
Recombinant form of Platlet II Repligen Clinical Factor 4 Partners
Vitakin Antibody antagonist of alpha-v- II MedImmune beta-3 ingrins
Volociximab Antibody antagonist of alpha-v- II Biogen Idec/Protein
beta-3 ingrins Design Labs 2- Estrogen metabolite that inhibits II
EntreMed methoxyestradiol HIF1 a translation AP-23573 Inhibits mTOR
II Ariad Pharmaceuticals Cancertinib TKI that inhibits EGFR II
Pfizer Actimid Thalomid derivative II Celgene Combretastatin
Tubulin destabilizing agent II Oxigene A4 prodrug Endo Tag 1
Antineovasculature agent, II Medigene formation of paclitaxel
encapsulated in positively charged liposomes Enzastaurin Protein
kinase C-beta inhibitor II Eli Lilly Ceflatonin Induces apoptosis
II ChemGenex Pharmaceuticals Silipide A complex of silybin and II
Indena phosphatidiylcholine INGN-241 Gene therapy based upon the II
Introgen mda-7 gene coding for IL-24 Therapeutics OSI-461 Inhibits
cGMP II OSI phospodiesterase Pharmaceuticals Patupilone A
non-taxane microtubule II Novartis stabilizing agent Squalamine
Blocks multiple angiogenic II Genaera cofactors Tacedinaline
Cystostatic histone II Pfizer deacetylation inhibitor UCN-01
Inhibitor of serine-threonine II NCI kinases, including protein
kinase C UK-356202 Urokinase-like plasminogen II Pfizer
activator
TABLE-US-00025 TABLE 6 Combination therapies for use in oncology
ABVD Doxorubcin, Bleomycin, Vinblastine, and Dacarbazine AC
Doxorubicin and Cyclophosphamide BEP Bleomycin, Etoposide and
Cisplatin CAF Cyclosphosphamide, Doxorubicin and 5-Fluorouracil
(5FU) CAV Cyclophosphamide, Doxorubicin, Vincristine Carboplatin-
Carboplatin and Etoposide Etoposide ChIVPP Chlorambucil,
Vinblastine, Procarbazine, and Prednisolone CHOP Cyclophosphamide,
Doxorubicin, Vincristine, and Prednisolone CHOP-R Cyclophosphamide,
Doxorubicin, Vincristine, Prednisolone, and Rituximab CMF
Cyclophosphamide, Methotrexate and 5FU CVAMP Cyclophosphamide,
Doxorubicin, Vincristine, and Methylprednisolone De Gramont 5FU and
leucovorin DHAP Dexamethasone, Cytarabine, and Cisplatin DAHP-R
Dexamethasone, Cytarabine, Cisplatin and Rituximab Doxorubicin-
Doxorubicin and Ifostamide Ifostamide EC Epirubicin and
Cyclophosphamide ECF Epirubicin, Cyclophosphamide, and 5FU ECMF
Epirubicin, Cyclophosphamide, Methotrexate, and 5FU EEX Epirubicin,
Oxaliplatin, and Capecitabine ECX Epirubicin, Cisplatin, and
Capecitabine ESHAP Etoposide, Methyl-prednisolone, Cytarabine and
Cisplatin FEC 5FU, Epirubicin, and Cyclophosphamide Gemcarbo
Gemcitabine and Carboplatin Gemcitabine- Gemcitabine and Cisplatin
Cisplatin Irinotecan-De Irinotecan, 5FU and Leucovorin Gramont MIC
Mitomycin, Ifosamide and Cisplatine MM Methotrexate and
Mitoxantrone MMM Methotrexate, Mitomycin, and Mitoxantrone MVP
Mitomycin, Vinblastine and Cisplatin FOLFOX 5FU, Oxilaplatin and
Leucovorin FOLFIRI 5FU, Leucovorin and Irinotecan Paclitaxel-
Paclitaxel and Carboplatin Carboplatin PmitCebo Prednisolone,
Mitoxantrone, Cyclophosphamide, Etoposide, Bleomycin and
Vincristine VAD Vincristine, Doxorubicin, and Dexamethasone VAPEC-B
Vincristine, Doxorubicin, Prednisolone, Etoposide,
Cyclosphosphamide, and Bleomycin Vinorelabine- Vinorelabine and
Cisplatin Cisplatin
[0924] The versatility of the invention is illustrated by the
following Examples, which illustrate typical embodiments of the
invention and are not limiting of the claims or specification in
any way.
EXAMPLES
Example 1
Synthesis of Exemplary Compounds
##STR00239##
[0925] Example 2
Cleavage from Resin of the Peptide Prepared as in Example 1
[0926] Cleavage of Peptide from Resin
##STR00240##
Example 3
Coupling to Linker of the Compound Prepared as in Example 2
[0927] Coupling of Linker
##STR00241##
Example 4
Synthesis of
##STR00242##
[0928] is provided in FIG. 12.
Example 5
Synthesis of
##STR00243##
[0929] is provided in FIG. 13.
Example 6
Synthesis of
##STR00244##
[0930] is provided in FIG. 14.
Example 7
Synthesis of
##STR00245##
[0931] is provided in FIG. 15.
Example 8
Synthesis of
##STR00246##
[0932] is provided in FIG. 16.
Example 9
Synthesis of
##STR00247##
[0933] is provided in FIG. 17.
Example 10
Synthesis of
##STR00248##
[0934] is provided in FIG. 18.
Example 11
Synthesis of
##STR00249##
[0935] is provided in FIG. 19.
Example 12
Synthesis of
##STR00250##
[0936] is provided in FIG. 20.
Example 13
Synthesis of
##STR00251##
[0937] is provided in FIG. 21.
Example 14
Synthesis of
##STR00252##
[0938] is provided in FIG. 22.
Example 15
Synthesis of
##STR00253##
[0939] is provided in FIG. 23.
Example 16
Synthesis of
##STR00254##
[0940] is provided in FIG. 24.
Example 17
Synthesis of
##STR00255##
[0941] is provided in FIG. 25. While this EXAMPLE uses the compound
of EXAMPLE 12, it could also sufficiently employ the compounds of
EXAMPLE 13. Further, while this EXAMPLE shows linking to the
N-terminus, the free acid on the left side of the compounds of
EXAMPLES 12 and 13 may also be linked to any nucleophilic side
chain on a peptide, such as the C, K, S, T or Y side chains. As is
also shown in this EXAMPLE, the Fmoc protected amino group on the
right side of the compounds of EXAMPLES 12 and 13 is used to link
to the recognition group, Y, via an amide bond.
Example 18
Synthesis of
##STR00256##
[0942] is provided in FIG. 26. While this EXAMPLE uses the compound
of EXAMPLE 15, it could also sufficiently employ the compounds of
EXAMPLES 14 and 16. Further, while this EXAMPLE shows linking to
the N-terminus, the free acid on the left side of the compounds of
EXAMPLES 14-16 may also be linked to any nucleophilic side chain on
a peptide, such as the C, K, S, T or Y side chains. As is also
shown in this EXAMPLE, the free acid on the right side of the
compounds of EXAMPLES 14-16 is used to link to the antibody
recognition group, Y, via an amide bond.
Example 19
Synthesis of
3-{2-[2-(2-{2-[2-(2-tert-Butoxycarbonyl-ethoxy)-ethoxyl]ethoxy}-ethoxy)-et-
hoxy]ethoxy}-propionic acid tert-butyl ester
##STR00257##
[0944] The title compound was prepared using a reported method (O.
Seitz and H. Kunz, J. Org. Chem. 62:813-826 (1997)). A small piece
of sodium metal was added to a solution of tetra(ethylene glycol)
(47.5 g, 244 mmol) in THF (200 ml) and stirred until the sodium was
dissolved completely. .sup.tButyl acrylate (94 g, 730 mmol) was
then added and stirring continued for 2 days at RT. Another batch
of .sup.tButyl acrylate (94 g, 730 mmol) was added and stirring
continued for another 2 days. The reaction mixture was neutralized
with a few drops of 1N HCl and concentrated under reduced pressure.
The residue was suspended in water and extracted with ethyl acetate
(3.times.150 ml). Combined organic layers were washed with brine
and dried over sodium sulfate. Evaporation of volatiles over
reduced pressure provided the crude product as colorless liquid
which was purified using a silica gel column (42 g, 51%).
Example 20
Synthesis of
3-{2-[2-(2-{2-[2-(2-Carboxy-ethoxy)-ethoxyl]ethoxy}-ethoxy)-ethoxyl]ethoxy-
}-propionic acid
##STR00258##
[0946] A solution of
3-{2-[2-(2-{2-[2-(2-tert-Butoxycarbonyl-ethoxy)-ethoxy]-ethoxyl}-ethoxy)--
ethoxy]-ethoxyl}-propionic acid tert-butyl ester (6 g, 18.6 mmol)
in anisole (20 ml) was cooled in an ice bath and trifluroacetic
acid (65 g) was added. After 3 hrs at RT volatiles were removed
under reduced pressure and the residue was partitioned between
ethyl acetate (50 ml) and 5% sodium bicarbonate solution. The
aqueous layer was acidified with 1 N HCl, saturated with NaCl and
then extracted with ethyl acetate (3.times.50 ml). Combined organic
layers were washed with brine and dried over sodium sulfate.
Removal of volatiles under the reduced pressure provided the
product as colorless liquid which solidified upon refrigeration
(3.8 g, 82%).
Example 21
Synthesis of
3-(2-{2-[2-(2-{2-[2-(4-{2-[2-(2-Methyl-[1,3]dioxolan-2-ylmethyl)-[1,3]diox-
olan-2-yl]-ethyl}-phenylcarbamoyl)-ethoxyl]ethoxy}-ethoxy)-ethoxyl]ethoxy}-
-ethoxy)-propionic acid
##STR00259##
[0948] Compound from EXAMPLE 20 (0.6 g, 1.8 mmol) was dissolved in
dichloromethane (10 ml) and
4-{2-[2-(2-Methyl-[1,3]dioxolan-2-ylmethyl)-[1,3]dioxolan-2-yl]-ethyl}-ph-
enylamine (0.3 g, 1.4 mmol) followed by EDCl (0.28 g, 1.8 mmol) was
added at RT. After 1 hr at RT the RM was washed with water and
dried over sodium sulfate. Evaporation of volatiles and
purification over silica gel column with 1 to 15% methanol in
dichloromethane provided title compound as gum (0.47 g, 32%).
Example 22
Synthesis of
4-{2-[2-(2-Methyl-[1,3]dioxolan-2-ylmethyl)-[1,3]dioxolan-2-yl]-ethyl}-phe-
nylamine
##STR00260##
[0950] A clean oven dried flask was charged with the
6-(4-nitro-phenyl)-hexane-2,4-dione (3.7 g, 15.72 mmol), dry
CH.sub.2Cl.sub.2 (20 ml) followed by bisTMS ethylene glycol (38.5
ml, 157.3 ml) were added to the flask and the resulting solution
was cooled to -5.degree. C. with stirring under argon. TMSOTf (300
.mu.l) was added to the reaction mixture and the solution was
stirred at -5.degree. C. for 6 h. Reaction was quenched with
pyridine (10 ml) and poured into sat. NaHCO.sub.3. The mixture was
extracted with EtOAc and the organic layer was washed with water,
brine, dried (Na.sub.2SO.sub.4) and concentrated to give a yellow
solid. The solid was triturated with hexanes to give a free flowing
pale yellow solid (3.5 g, 72%) which was dissolved in EtOAc (50 ml)
and hydrogenated on a Parr shaker starting with 50 psi of hydrogen
pressure. After two hours the reaction was filtered through a pad
of celite, the celite was washed thoroughly with
CH.sub.2Cl.sub.2/MeOH and combined organics were concentrated to
give title compound (1.46 g, 100%) as an oil that solidifies upon
standing.
Example 23
Synthesis of
Synthesis of 4-[4-(3,5-Dioxo-hexyl)-phenylcarbamoyl]-butyric acid
2,5-dioxo pyrrolidin-1-yl ester (10)
##STR00261##
[0951] Step 1: 6-(4-Nitro-phenyl)-hexane-2,4-dione (11)
[0952] To a reaction vessel (heat and vacuum dried and equipped
with a magnetic spin bar) was added tetrahydrofuran and lithium
diisopropylamide (2M heptane/ethylbenzene/tetrahydrofuran; 69.4 mL,
138.9 mmol). The solution cooled to -78.degree. C.
Pentane-2,4-dione (7.13 mL, 69.4 mmol) was added dropwise and the
solution stirred 30 minutes at -78.degree. C. 4-nitrobenzyl bromide
(15.0 g, 69.4 mmol) was added in one portion. The solution was
removed from the dry-ice/acetone bath, allowed to warm to room
temperature and stirred 16 hours. The solution was cooled to
approximately 0.degree. C. and the reaction quenched with 1M HCl.
Tetrahydrofuran was removed under reduced pressure. The crude
material was taken up into dichloromethane and washed with 1M HCl
and brine. The aqueous layers were again washed with
dichloromethane. The combined dichloromethane layers were dried
(Na.sub.2SO.sub.4) and removed under reduced pressure. Gradient
flash column chromatography (FCC) was performed using 5% to 15%
ethyl acetate/hexanes to afford title compound (8.5 g, 52%; yellow
solid). .sup.1H NMR (CDCl.sub.3): 8.14 (d, J=9.0 Hz, 2H), d, J=8.4
Hz, 2H), 5.45 (s, 1H), 3.06 (t, J=7.5 Hz, 2H), 2.64 (t, J=7.8 Hz,
2H), 2.04 (s, 3H).
Step 2: 4-[4-(3,5-Dioxo-hexyl)-phenylcarbamoyl]-butyric acid
(12)
[0953] 200 mL tetrahydrofuran, 6-(4-nitro-phenyl)-hexane-2,4-dione
(8.0 g, 34.0 mmol) and dihydro-pyran-2,6-dione (3.88 g, 34.0 mmol)
were added to a reaction vessel. The reaction vessel was purged
three times with argon. Approximately 200 mg palladium (10 wt % on
activated carbon) was added. The reaction vessel was purged again
with argon and excess hydrogen introduced via a balloon. Solution
stirred 16 hours at room temperature. Hydrogen removed under
reduced pressure and catalyst removed by filtration through celite.
Tetrahydrofuran removed under reduced pressure to afford title
compound (10.5 g, 97%, yellow solid).
Step 3: 4-[4-(3,5-Dioxo-hexyl)-phenylcarbamoyl]-butyric acid
2,5-dioxo pyrrolidin-1-yl ester (10)
[0954] To a reaction vessel (heat and vacuum dried and equipped
with a magnetic spin bar) was added
4-[4-(3,5-dioxo-hexyl)-phenylcarbamoyl]-butyric acid (10.53 g, 33.0
mmol), N-hydroxysuccinimide (3.8 g, 33.0 mmol) and
1-[3-(dimethylamino) propyl]-3-ethylcarbodiimide hydrochloride (6.3
g, 33.0 mmol) and dichloromethane (250 mL). The solution was
stirred under nitrogen at room temperature for 16 hours then washed
with 10% citric acid, brine and dried (Na.sub.2SO.sub.4).
Dichloromethane was removed under reduced pressure. FCC with 70%
ethyl acetate/hexanes gave title compound (7.4 g, yellow solid,
54%). .sup.1H NMR (CDCl.sub.3): 7.87 (s, 1H), 7.43 (d, J=8.4 Hz,
2H), 7.12 (d, J=8.4 Hz, 2H), 5.46 (s, 1H), 2.89 (t (& m), J=8.1
Hz (for the t), 7H), 2.73 (t, J=6.0 Hz, 2H), 2.56 (t, J=7.2 Hz,
2H), 2.47 (t, J=6.9 Hz, 2H), 2.21 (p, J=6.6 Hz, 2H), 2.04 (s,
3H).
Example 24
Synthesis of
Synthesis of
3-{2-[2-(2-{4-[4-(3,5-Dioxo-hexyl)-phenylcarbamoyl]-butyrylamino}-ethoxy)-
-ethoxyl]ethoxy}-propionic acid 2,5-dioxo-pyrrolidin-1-yl ester,
(20)
##STR00262##
[0955] Step 1: 3-{2-[2-(2-Hydroxy-ethoxy)-ethoxy]-ethoxy}-propionic
acid tert-butyl ester
[0956] Na metal (catalytic) was added to a stirring solution of
acrylic acid tert-butyl ester (6.7 mL, 46 mmol), and
2-[2-(2-hydroxy-ethoxy)-ethoxy]-ethanol (20.7 g, 138 mmol) in THF
(100 mL) at 0.degree. C. and the mixture was stirred overnight.
Solvent was removed and the remaining oil dissolved in EtOAc (100
mL). The organic layer was washed with water (3.times.50 mL), and
dried over Na.sub.2SO.sub.4 and the solvent removed in vacuo to
give an oil which corresponds to the title compound that would be
used as is for the next step. (M+1)=279.
Step 2:
3-{2-[2-(2-Tosylsulfonyloxy-ethoxy)-ethoxyl]ethoxy}-propionic acid
tert-butyl ester
[0957] Tosyl chloride (22.3 g, 117 mmol) was added in portions to a
stirring solution of
3-{2-[2-(2-hydroxy-ethoxy)-ethoxy]-ethoxy}-propionic acid
tert-butyl ester (16.3 g, 58.6 mmol) and pyridine 60 mL in (240 mL)
and the mixture was stirred overnight. The reaction was quenched
with water (300 mL) and the organic layer was separated. The
aqueous layer was extracted with CH.sub.2Cl.sub.2 (2.times.100 mL).
The combined organic layer was washed with HCl (1N, 100 mL), water
(100 mL), and dried over Na.sub.2SO.sub.4 and the solvent was
removed in vacuo to give an oil which corresponds to the title
compound that would be used as is for the next step. (M+1)=433.
Step 3: 3-{2-[2-(2-Amino-ethoxy)-ethoxy]-ethoxy}-propionic acid
tert-butyl ester
[0958] NaN.sub.3 (35 g, 538 mmol) was added to a stirring solution
of 3-{2-[2-(2-tosylsulfonyloxy-ethoxy)-ethoxy]-ethoxy}-propionic
acid tert-butyl ester (20 g, 46 mmol) in DMF (150 mL) and the
reaction was stirred overnight. Reaction was diluted with water
(200 mL) and extracted with EtOAc (4.times.100 mL). The organic
layer was washed with water (100 mL) and brine (100 mL) and dried
over Na.sub.2SO.sub.4. The solvent was removed in vacuo to give an
oil. Column chromatography EtOAc/Hex (1:4) gave an oil which
corresponds to the
3-{2-[2-(2-azido-ethoxy)-ethoxy]-ethoxy}-propionic acid tert-butyl
ester, (M+1)=304. This oil was hydrogenated using Pd (5% on carbon)
in EtOAc under hydrogen (1 atm.) over 3 days. The catalyst was
removed by filtration and solvent removed in vacuo to give an oil
corresponding to the title compound, (M+1)=278.
Step 4:
3-{2-[2-(2-{4-[4-(3,5-Dioxo-hexyl)-phenylcarbamoyl]-butyrylamino}--
ethoxy)-ethoxyl]ethoxy}-propionic acid tert-butyl ester
[0959] A solution of
4-[4-(3,5-dioxo-hexyl)-phenylcarbamoyl]-butyric acid
2,5-dioxo-pyrrolidin-1-yl ester (1.5 g, 3.6 mmol),
3-{2-[2-(2-amino-ethoxy)-ethoxy]-ethoxy}-propionic acid tert-butyl
ester (1.0 g, 3.6 mmol) and DIEA (1.3 L, 7.2 mmol) in
CH.sub.2Cl.sub.2 (10 mL) was stirred at rt overnight. The solvent
was removed in vacuo and the residual oil purified using column
chromatography EtOAc/MeOH (95:5) to give the title compound as a
transparent oil, (M+1)=579.
Step 5:
3-{2-[2-(2-{4-[4-(3,5-Dioxo-hexyl)-phenylcarbamoyl]-butyrylamino}--
ethoxy)-ethoxy]-ethoxy}-propionic acid 2,5-dioxo-pyrrolidin-1-yl
ester
[0960]
3-{2-[2-(2-{4-[4-(3,5-Dioxo-hexyl)-phenylcarbamoyl]-butyrylamino}-e-
thoxy)-ethoxy]-ethoxyl}-propionic acid tert-butyl ester (400 mg,
0.692 mmol) was dissolved in TFA/CH.sub.2Cl.sub.2 (1:1, 3 mL) and
the mixture stirred overnight. The solvent was removed to give an
oil as the acid intermediate. This oil was dissolved in
CH.sub.2Cl.sub.2 (4 mL) containing DIEA (569 L, 3.09 mmol),
N-hydroxysuccinimide (119 mg, 1.03 mmol) and EDC (197 mg, 1.0 mmol)
and the mixture stirred over the night. The solvent was removed and
the residual oil was purified using column chromatography
EtOAc/MeOH (95:5) to give an oil as the title compound,
(M+1)=620.
Example 25
Synthesis of AA Targeting Compound
[0961] Compound of EXAMPLES 17 or 18 can be linked to antibody 38C2
by the following procedure: One mL antibody 38C2 in phosphate
buffered saline (10 mg/mL) is added to 12 .mu.L of a 10 mg/mL stock
solution of AA targeting agent and the resulting mixture maintained
at room temperature for 2 hours prior to use.
Example 26
Synthesis of
##STR00263##
[0962] is provided in FIG. 27. While this EXAMPLE uses the compound
of EXAMPLE 12, it could also sufficiently employ the compounds of
EXAMPLE 13.
Example 27
[0963] C. Rader, et al., J. Mol. Biol. 332:889-899 (2003) details
one method of making h38c2. The following details the results,
materials and methods in this reference.
[0964] Humanization Human V.kappa. gene DPK-9 and human J.kappa.
gene JK4 were used as frameworks for the humanization of the kappa
light chain variable domain, and human VH gene DP-47 and human
J.sub.H gene J.sub.H4 are used as frameworks for the humanization
of the heavy chain variable domain of m38C2. All complementarity
determining region (CDR) residues as defined by Kabat et al., as
well as defined framework residues in both light chain and heavy
chain variable domain, were grafted from m38C2 onto the human
framework. The selection of grafted framework residues may be based
on the crystal structure of mouse mAb 33F12 Fab (PDB 1AXT). mAb
33F12 Fab shares a 92% sequence homology with m38c2 in the variable
domains and identical CDR lengths. Furthermore, both 33F12 and
m38C2 have similar catalytic activity. Framework residues consisted
of five residues in the light chain and seven residues in the heavy
chain (FIG. 7A) and encompassed the residues that are likely to
participate directly or indirectly in the catalytic activity of
m38C2. These include the reactive lysine of m38C2, Lys.sup.H93,
which is positioned in framework region 3 (FR3) of the heavy chain.
Six residues, Ser.sup.H35, Val.sup.H37, Trp.sup.H47, Trp.sup.H103,
and Phe.sup.L98, which are conserved between mouse mAbs 33F12 and
38C2, are within a 5-.ANG. radius of the .epsilon. amino group of
Lys.sup.H93. These residues were also conserved in the
humanization. Lys.sup.H93 lies at the bottom of a highly
hydrophobic substrate binding sites of mouse mAbs 33F12 and 38C2.
In addition to CDR residues, a number of framework residues line
this pocket. Among these, Leu.sup.L37, Gln.sup.L42, Ser.sup.L43,
Val.sup.L85, Phe.sup.L87, Val.sup.H5, Ser.sup.H40, Glu.sup.H42,
Gly.sup.H88, Ile.sup.H89, and Thr.sup.H94 were grafted onto the
human framework.
[0965] Expression By fusing the humanized variable domains to human
constant domains C.sub..kappa. and C.sub..gamma.11, h38C2 was
initially generated as Fab expressed in E. coli. Next, h38c2 IgG
was formed from h38c2 Fab using the PIGG vector engineered for
human IgG1 expression in mammalian cells. Supernatants from
transiently transfected human 293T cells were subjected to affinity
chromatography on recombinant protein A, yielding approximately 1
mg/L h38C2 IgG1. Purity was established by SDS-PAGE followed by
Coomassie blue staining.
[0966] --Diketone Compounds--
##STR00264##
[0967] The enaminone formed by the covalent addition of a
.beta.-diketone with m38c2 has a characteristic UV absorbance at
.lamda..sub.max=318 nm. Like m38C2 IgG, h38C2 IgG showed the
characteristic enaminone absorbance after incubation with
.beta.-diketone. As a negative control, recombinant human
anti-HIV-1 gp120 mAb b12 with the same IgG1 isotype as h38C2 but
without reactive lysine, did not reveal enaminone absorbance after
incubation with .beta.-diketone 2. For a quantitative comparison of
the binding of 6-diketones to m38C2 and h38C2, the authors used a
competition ELISA. The antibodies were incubated with increasing
concentrations of .beta.-diketones 2 and 3 and assayed against
immobilized BSA-conjugated .beta.-diketone 1. The apparent
equilibrium dissociation constants were 38 .mu.M (m38C2) and 7.6
.mu.M (h38C2) for .beta.-diketone 2 and 0.43 .mu.M (m38C2) and 1.0
.mu.M (h38C2) for .beta.-diketone 3, revealing similar
.beta.-diketone binding properties for mouse and humanized antibody
(FIG. 6).
Molecular Modeling
[0968] A molecular model of h38C2 Fab was constructed by homology
modeling using the crystal structure of a related aldolase
antibody, mouse 33F12 Fab (Protein Data Bank ID: 1AXT), as a
template. The crystal structure of mouse 33F12 Fab was previously
determined at a resolution of 2.15 .ANG...sup.4 Alignment of mouse
33F12 and 38C2 amino acid sequences using the HOMOLOGY module
within INSIGHT II software (Accelrys) confirmed that both sequences
are highly homologous. They differ from each other by 19 out of 226
amino acids in the two variable domains, and their CDRs share the
same lengths. In addition to the high sequence homology, both
structures exhibit considerable structural similarity, as observed
by a low-resolution crystal structure of 38C2. Residues in the
model were mutated to conform to the h38C2 amino acid sequence and
sidechains were placed based on standard rotamers. This model was
then minimized with the DISCOVER module in INSIGHT II using 100
steps each of steepest descent minimization followed by conjugate
gradient minimization.
Construction of h38C2 Fab
[0969] The sequences of the variable light and heavy chain domains
of m38C2 (SEQ ID NOs:15 and 16, respectively) as well as the
sequences of human germline sequences DPK-9 (SEQ ID NO:17), JK4
(SEQ ID NO:18), DP-47 (SEQ ID NO:19), and JH4 (SEQ ID NOs:20, 187
and 188) (V BASE; http://vbase.mrc-cpe.cam.ac.uk/) were used to
design overlapping oligonucleotides for the synthetic assembly of
humanized V.sub..kappa. and V.sub.H, respectively. N-glycosylation
sites with the sequence NXS/T as well as internal restriction sites
HindIII, XbaI, SacI, ApaI, and SfiI were avoided. PCR was carried
out by using the Expand High Fidelity PCR System (Roche Molecular
Systems). The humanized V.sub..kappa. oligonucleotides were: L
flank sense (Rader, C., Ritter, G., Nathan, S., Elia, M., Gout, I.,
Junbluth, A. A., J. Biol. Chem. 275: 13668-13676 (2000)) (sense
5'-GAGGAGGAGGAGGAGGGCCCAGGCGGCCGAGCTCCAGATGACCCAGTCTCTCCA-3' SEQ ID
NO:179); h38C2L1 (sense;
5'-GAGCTCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGTGACCGCGT
CACCATCACTTG-3') (SEQ ID NO:1); h38C2L2 (antisense;
5'-ATTCAGATATGGGCTGCCATAAGTGTGCAGGAGGCTCTGACTGGAGCGGCAAGTG
ATGGTGACGCGGTC-3') (SEQ ID NO:2); h38C2L3 (sense;
5'-TATGGCAGCCCATATCTGAATTGGTATCTCCAGAAACCAGGCCAGTCTCCTAAGCT
CCTGATCTAT-3') (SEQ ID NO:3); h38C2L4 (antisense;
5'-CTGAAACGTGATGGGACACCACTGAAACGATTGGACACTTTATAGATCAGGAGCTT
AGGAGACTG-3') (SEQ ID NO:4); h38C2L5 (sense;
5'-AGTGGTGTCCCATCACGTTTCAGTGGCAGTGGTTCTGGCACAGATTTCACTCTCAC
CATCAGCAGTCTGCAACCTGAAGATTTTGCAGTG-3') (SEQ ID NO:5); h38C2L6
(antisense;
5'-GATCTCCACCTTGGTCCCTCCGCCGAAAGTATAAGGGAGGTGGGTGCCCTGACTA
CAGAAGTACACTGCAAAATCTTCAGGTTGCAG-3') (SEQ ID NO:6); L antisense
flank (C. Rader et al., J. Biol. Chem. 275:13668-13676 (2000))
(antisense5'-GACAGATGGTGCAGCCACAGTTCGTTTGATCTCCACCTTGGTCCCTCC-3'
SEQ ID NO:180). The humanized V.sub.H oligonucleotides were: H
flank sense (C. Rader et al., J. Biol. Chem. 275:13668-13676
(2000))(sense
5'-GCTGCCCAACCAGCCATGGCCGAGGTGCAGCTGGTGGAGTCTGGGGGA-3' SEQ ID
NO:181); h38C2H1 (sense;
5'-GAGGTGCAGCTGGTGGAGTCTGGCGGTGGCTTGGTACAGCCTGGCGGTTCCCTG
CGCCTCTCCTGTGCAGCCTCTGGCT-3') (SEQ ID NO:7); h38C2H2 (antisense;
5'-CTCCAGGCCCTTCTCTGGAGACTGGCGGACCCAGCTCATCCAATAGTTGCTAAAG
GTGAAGCCAGAGGCTGCACAGGAGAG-3') (SEQ ID NO:8); h38C2H3 (sense;
5'-TCTCCAGAGAAGGGCCTGGAGTGGGTCTCAGAGATTCGTCTGCGCAGTGACAACT
ACGCCACGCACTATGCAGAGTCTGTC-3') (SEQ ID NO:9); h38C2H4 (antisense;
5'-CAGATACAGCGTGTTCTTGGAATTGTCACGGGAGATGGTGAAGCGGCCCTTGACA
GACTCTGCATAGTGCGTG-3') (SEQ ID NO:10); h38C2H5 (sense;
5'-CAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGCGCGCCGAGGACACG
GGCATTTATTACTGTAAAACG-3') (SEQ ID NO:11); h38C2H6 (antisense;
5'-TGAGGAGACGGTGACCAGGGTGCCCTGGCCCCAGTAGCTGAAACTGTAGAAGTAC
GTTTTACAGTAATAAATGCCCGTG-3') (SEQ ID NO:12); H flank antisense (C.
Rader et al., J. Biol. Chem. 275:13668-13676 (2000))(antisense
5'-GACCGATGGGCCCTTGGTGGAGGCTGAGGAGACGGTGACCAGGGTGCC-3' SEQ ID
NO:182). Following assembly, humanized V.sub..kappa. and V.sub.H
were fused to human C.sub..kappa. and CO, respectively, and the
resulting light chain and heavy chain fragment were fused and
SfiI-cloned into phagemid vector pComb3X as described (C. Rader et
al, J. Biol. Chem. 275:13668-13676 (2000); C. F. Barbas 3.sup.rd et
al., Phage Display: A laboratory manual, Cold Spring Harbor
Laboratory, Cold Spring Harbor N.Y. (2001)). To enrich for clones
with the correct h38C2 sequence, Fab were displayed on phage and
selected by one round of panning against the immobilized
.beta.-diketone 1 (JW) conjugated to BSA. Soluble Fab were produced
from single clones and tested for binding to immobilized JW-BSA by
ELISA using donkey anti-human F(ab').sub.2 polyclonal antibodies
conjugated to horseradish peroxidase (Jackson ImmunoResearch
Laboratories) as secondary antibody. Light chain and heavy chain
encoding sequences of positive clones were analyzed by DNA
sequencing using the primers OMPSEQ (5'-AAGACAGCTATCGCGATTGCAG-3'
SEQ ID NO:183) and PELSEQ (5'-CTATTGCCTACGGCAGCCGCTG-3' SEQ ID
NO:184) (C. F. Barbas 3.sup.rd et al., Phage Display: A laboratory
manual, Cold Spring Harbor Laboratory, Cold Spring Harbor N.Y.,
(2001)), respectively, to confirm the assembled V.sub..kappa. and
V.sub.H sequences of h38C2.
Construction, Production, and Purification of h38C2 IgG1
[0970] The recently described vector PIGG (C. Rader et al, FASEB
J., 16:2000-2002 (2002)) was used for mammalian expression of h38C2
IgG1. The mammalian expression vector PIGG-h38c2 is illustrated in
FIG. 23. The 9 kb vector comprises heavy chain 1 and light chain
expression cassettes driven by a bidirectional CM promoter
construct. Using primers PIGG-h38C2H (sense;
5'-GAGGAGGAGGAGGAGGAGCTCACTCCGAGGTGCAGCTGGTGGAGTCTG-3') (SEQ ID
NO:13) and GBACK (5'-GCCCCCTTATTAGCGTTTGCCATC-3' SEQ ID NO:185) (C.
F. Barbas 3.sup.rd et al, Phage Display: A laboratory manual, Cold
Spring Harbor Laboratory, Cold Spring Harbor N.Y. (2001)), the VH
coding sequence from h38C2 Fab in phagemid vector pComb3X was
amplified, digested with SacI and ApaI, and cloned into the
appropriately digested vector PIGG. Using primers PIGG-h38C2L
(sense: 5'-GAGGAGGAGGAGGAGAAGCTTGTTGCTCTGGATCTCTGGTGCCTACGGGGAGCTC
CAGATGACCCAGTCTCC-3') (SEQ ID NO:14) and LEADB
(5'-GCCATGGCTGGTTGGGCAGC-3' SEQ ID NO:186) ((C. F. Barbas 3.sup.rd
et al, Phage Display: A laboratory manual, Cold Spring Harbor
Laboratory, Cold Spring Harbor N.Y. (2001)) the light chain coding
sequence from h38C2 Fab in phagemid vector pComb3X was amplified,
digested with HindIII and XbaI, and cloned into the appropriately
digested vector PIGG that already contained the h38C2 heavy chain.
Intermediate and final PIGG vector constructs were amplified in E.
coli strain SURE (Stratagene) and prepared with the QIAGEN Plasmid
Maxi Kit. h38C2 IgG1 were produced from the prepared final PIGG
vector construct by transient transfection of human 293T cells
using Lipofectamine 2000 (Invitrogen). Transfected cells were
maintained in GIBCO 10% ultra-low IgG (<0.1%) FCS (Invitrogen)
in RPMI 1640 (Hyclone) for 2 weeks. During this time, the medium
was collected and replaced three times. The collected medium was
subjected to affinity chromatography on a recombinant Protein A
HiTrap column (Amersham Biosciences). This purification step
yielded 2.45 mg h38C2 IgG1 from 2,300 mL collected medium as
determined by measuring the optical density at 280 nm using an
Eppendorf BioPhotometer. Following dialysis against PBS in a
Slide-A-Lyzer 10K dialysis cassette (Pierce), the antibody was
concentrated to 760 .mu.g/mL using an Ultrafree-15 Centrifugal
Filter Device (UFV2BTK40; Millipore), and sterile filtered through
a 0.2-.mu.m Acrodisc 13 MM S-200 Syringe Filter (Pall). The final
yield was 2.13 mg (87%). Purified h38C2 IgG1 was confirmed by
nonreducing SDS-PAGE followed by Coomassie Blue staining.
Enaminone Formation
[0971] Antibody (h38C2 IgG1 or b12 IgG1) was added to
.beta.-diketone 2 to a final concentration of 25 .mu.M antibody
binding site and 125 .mu.M .beta.-diketone. This mixture was
incubated at room temperature for 10 minutes before a UV spectrum
was acquired on a SpectraMax Plus 384 UV plate reader (Molecular
Devices) using SOFTmax Pro software (version 3.1.2).
Binding Assays
[0972] Unless noted otherwise, all solutions were phosphate
buffered saline (pH 7.4). A 2.times. solution of either
.beta.-diketone 2 or 3 (50 .mu.L) was added to 50 .mu.L of the
antibody (either h38C2 or m38C2) and allowed to incubate at
37.degree. C. for 1 hr. Solutions were mixed by pipetting. Final
concentrations of antibody were 0.4 to 8 nM antibody binding site,
and final concentrations of .beta.-diketones 2 and 3 were 10.sup.-9
to 10.sup.-2M and 10.sup.-10 to 10.sup.-4M, respectively. Each well
of a Costar 3690 96-well plate (Corning) was coated with 100 ng of
the BSA conjugate of .beta.-diketone 1 in TBS. Wells were then
blocked with 3% (w/v) BSA in TBS. Then, 50 .mu.L of the
antibody/.beta.-diketone mixture was added, followed by 50 .mu.L of
a 1:1,000 dilution of either goat anti-human Fc IgG polyclonal
antibodies (Pierce) or rabbit anti-mouse Fc IgG polyclonal
antibodies (Jackson ImmunoResearch Laboratories) conjugated to
horseradish peroxidase. This was followed by 50 .mu.L ABTS
substrate solution. Between each addition, the plate was covered,
incubated at 37.degree. C. for 1 hr, and then washed five times
with deionized H.sub.2O. The absorbance at 405 nm was monitored as
described above until the reaction with no .beta.-diketone reached
an appropriate value (0.5<A.sub.405<1.0). For each well, the
fractional inhibition of ELISA signal (v.sub.i) was calculated
using equation i:
v.sub.i=(A.sub.o-A.sub.i)/(A.sub.o) (i)
where A.sub.o is the ELISA absorbance obtained in the absence of
.beta.-diketone and A.sub.i is the absorbance obtained in the
presence of .beta.-diketone. For monovalent binding proteins, the
fraction of antibody bound to soluble .beta.-diketone (f) is equal
to v. However, the IgG antibody is bivalent, and the ELISA signal
is inhibited only by the presence of doubly liganded antibody and
not by monovalent binding. Therefore, the Stevens correction for a
bivalent antibody was used:
f.sub.i=(v.sub.i).sup.1/2 (ii)
[0973] The following relationship was used to determine the
apparent equilibrium dissociation constant (modified from [ref.
37]):
f.sub.i=f.sub.min+(f.sub.max-f.sub.min)(1+K.sub.D/a.sub.0).sup.-1
(iii)
where a.sub.0 corresponds to the total .beta.-diketone
concentration, K.sub.D is the equilibrium dissociation constant,
and f.sub.min and f.sub.max represent the experimentally determined
values when the antibody binding sites are unoccupied or saturated,
respectively. Because this equation is only valid when the K.sub.D
values are at least 10.times. higher than the antibody
concentration, it was verified that the K.sub.D values determined
from equation iii met this criterion. Data were fit using a
nonlinear least-squares fitting procedure of KaleidaGraph (version
3.0.5, Abelbeck software) with K.sub.D, f.sub.max, and f.sub.min as
the adjustable parameters and normalized using equation iv:
f.sub.norm=(f.sub.i-f.sub.min)/(f.sub.max-f.sub.min) (iv)
Example 28
[0974] The ability of Ang-2 binding compounds to interact with
Ang-2 was measured by competition with Tie-2.
[0975] For competitive ELISA, human angiopoietin-2 protein and
Tie-2-Fc (R&D Systems) were reconstituted without carrier
protein. Mouse anti-human Tie-2 (Pharmingen) was used as the
primary antibody and goat anti-mouse-IgG1-HRP (Pierce) was used as
the secondary antibody. TMB substrate from Pierce was used.
[0976] High-binding half-well plates were coated with Ang-2 (100
ng/well) in 50 .mu.l PBS and incubated at 4.degree. C. overnight.
Plates were washed three times with washing buffer (0.1% Tween 20,
PBS, pH 7.4) and blocked with Superblock (Scytek), 100 at room
temperature ("RT") for 1 hour. After removing the blocking
solution, 50 .mu.l of an Ang-2 binding peptide compound (1 uM and
5.times. serial dilution) in the presence of 0.25 nM hTie-2-Fc
using Superblock as diluent was added and incubated at RT for 2
hours. Plates were washed 3 times with washing buffer. Then, 50 ul
of 0.1 ug/ml mouse anti-human Tie-2 diluted in Superblock added and
incubated at RT for 1 hour. Following incubation, 50 ul of 1:5,000
dilution of goat anti-mouse IgG-HRP in Superblock was then added
and incubated at RT for one hour. After washing 3 times, 50 .mu.l
(25 .mu.l TMB+25 .mu.l H.sub.2O.sub.2) was added, and incubated for
3-5 minutes. Color development was monitored and stopped with 25
.mu.l of 2 M H.sub.2SO.sub.4. OD450 nm with a correction wavelength
of 540 nm was measured. IC50 values (50% inhibition of Ang-2-Tie-2
binding) were calculated using non-linear Sigmoidal dose-response
curve fitting function in the Prism 4 software (GraphPad).
[0977] For reverse competition ELISA, human Tie-2-Fc,
angiopoietin-2 protein, biotinylated anti-human Ang-2 antibody, and
streptavidin HRP (R&D Systems) and TMB substrate from Pierce
were used.
[0978] High-binding half-well plates were coated with Tie-2-Fc (50
ng/well) in 50 .mu.l PBS and incubated at 4.degree. C. overnight.
Plates were washed three times with washing buffer (0.1% Tween 20,
PBS, pH 7.4) and blocked with Superblock, 150 .mu.l/well at RT for
1 hour. Plates were washed three times. Following washing, 50 .mu.l
of an Ang-2 binding peptide compound (50 nM, 5.times. serial
dilution) in the presence of 50 ng/ml (0.83 nM) Ang-2 in Superblock
were added and incubated at RT for 1 hour. Plates were washed 3
times, 50 .mu.l of 1 .mu.g/mlbiotinylated anti-Ang-2 detection
antibody in Superblock was added and incubated at RT for 2 hours.
Plates were washed 3 times, and 50 .mu.l of streptavidin HRP (1:200
dilution in Superblock) was added at RT for 20 minutes. Plates were
washed 3 times, and 50 .mu.l (25 .mu.l TMB+25 .mu.l H.sub.2O.sub.2)
substrate solution was added and incubated for 20-30 minutes. Color
development was stopped with 25 .mu.l of 2 M H.sub.2SO.sub.4. OD450
nm with a correction wavelength of 540 nm was measured. IC50 values
(50% inhibition of Ang-2-Tie-2 binding) were calculated using
non-linear Sigmoidal dose-response curve fitting function in the
Prism 4 software.
[0979] IC50 values for exemplary Ang-2 binding peptide compounds as
determined by competitive ELISA are presented in Table 7. IC50
values are provided for the targeting peptide plus linker (as shown
in FIG. 3, or FIG. 2 for compounds 24 and 25) (T) and the targeting
peptide linked to an antibody (P) via the linker of FIG. 3, unless
otherwise specified. In Table 7, compounds of SEQ ID NOs: 21-23 are
Ang-2 binding peptides alone, not conjugated to either a linker or
linker-antibody; compounds of SEQ ID NOs:65 and 66 are Ang-2
binding peptides conjugated to a linker-antibody, where the linker
is 4P ("4" PEG) and has the structure of the linker shown in FIG.
2; and compounds 26-63 are Ang-2 binding peptides conjugated to a
linker-antibody, where the linker is OP ("0" PEG) and has the
structure of the linker shown in FIG. 3. Compounds 24-63 were
conjugated to humanized aldolase antibody h38c2 and the linker
structures shown in FIG. 2 (4P) and FIG. 3 (OP) when obtaining the
data shown below, except where otherwise indicated. All compounds
of the invention shown in the Tables were capped with an acyl group
at the N-terminus and an amino group at the C-terminus, except
where otherwise indicated (e.g. compounds 26, 49, 50, 51 and
52).
[0980] In Table 7, amino acid sequences of peptide compounds are
shown with the position of linker OP or 4P indicated in parentheses
following the internal amino acid residue to which the linker is
attached. For compound of SEQ ID NO:67, the N-terminal "OP" linker
is indicated at the beginning of the peptide sequence.
[0981] For example, compound 24 in Table 7 has the following
sequence: QAcKY QPL DEL DK(4P)T LYD QFM LQQ G (SEQ ID NO:65). In
this example, the second amino acid residue is epsilon acyl lysine,
followed by tyrosine, and the linking position (in this case a 4P
linker) is the lysine residue 11, followed by threonine. Also,
compound 52 has the following sequence: (Amido 2-PEG)QAcKY QPL DEL
DK(OP)T LYD QFMLQQ G (SEQ ID NO:93). In this case, the N-terminal
glutamine residue is capped by an amido-2-PEG group, the second
amino acid residue is epsilon acyl lysine, and the OP linker is
attached to lysine residue 11.
[0982] Tables 7 and 8 also show half-life (T %) and "screening"
half life (results in parentheses): this is essentially an
alternative method of determining T %, based on a shorter test
period. For "Screening" T %, test compounds were intravenously
administered into male Swiss Webster mice. Blood samples were taken
from 4 mice per time point via retroorbital sinus bleed at the
following time points: 0.08, 5, and 32 hours. Blood level of test
compounds were determined by ELISA. The data were reported as the
percentage of test compounds at 32 hours versus 5 min. Normal T1/2
was calculated in a similar fashion, after undergoing additional
data analysis using WinNonlin version 4.1 (Pharsight Corporation).
The data was fit to a model based upon the shape of the curve (i.e.
a bi-exponential decline will be fit to a two compartment model,
etc.) The criteria for best fit (i.e. lowest % CV) was based on
iterative re-weighted least squares.
TABLE-US-00026 TABLE 7 T1/2 Ang-2 T Ang-2 P (Sc) Compound Sequence
IC50 nM IC50 nM hours 21 QKY QPL DEL DKT LYD QFM LQQ G 25.81 (SEQ
ID NO: 21) 22 Q(AcK)Y QPL DEL DKT LYD QFM LQQ G 41.9 (SEQ ID NO:
22) 23 QNY QPL DEL DKT LYD QFM LQQ G 27.83 (SEQ ID NO: 23) 24
Q(AcK)Y QPL DEL DK(4P)T LYD QFM LQQ G 206.7 17.4 104 (SEQ ID NO:
65) (27) 25 QNY QPL DEL DK(4P)T LYD QFM LQQ G 300.3 40.456 (SEQ ID
NO: 66) 26 (0P)QKY QPL DEL DKT LYD QFM LQQ G 32.3 58.77 (7) (SEQ ID
NO: 67) 27 QKY QPL DEL DK(0P)T LYD QFM LQQ G 29 55.876 117 (SEQ ID
NO: 68) (32) 28 QNY QPL DEL DK(0P)T LYD QFM LQQ G 175 28.63 100
(SEQ ID NO: 69) (57) 29 Q(AcK)Y QPL DEL DK(0P)T LYD QFM LQQ G 83.3
13.195 77 (33) (SEQ ID NO: 70) 30 Q(AcK)Y QPL DEK(0P) D(AcK)T LYD
QFM LQQ G 279 65.4 (17) (SEQ ID NO: 71) 31 Q(AcK)Y QPL DEL DET
K(0P)YD QFM LQQ G >5K >1000 (SEQ ID NO: 72) 32 Q(AcK)Y QPL
DEL D(AcK)T K(0P)YD QFM LQQ G >5K >1000 (SEQ ID NO: 73) 33
Q(AcK)Y QPL DEK(0P) DET LYD QFM LQQ G 369.6 88.81 (SEQ ID NO: 74)
34 Q(AcK)Y QD(HP)L DEL DK(0P)T LYD QFM LQQ G 224.5 19.75 (31) (SEQ
ID NO: 75) 35 Q(AcK)Y Q(HP)L DEL DK(0P)T LYD QFM LQQ G 121 16.68
(SEQ ID NO: 76) 36 Q(AcK)Y QPL DEL DK(0P)T IYD QFM LQQ G 2410 152.9
(SEQ ID NO: 77) 37 Q(AcK)Y QPL DEI DK(0P)T LYD QFM LQQ G 1273 80.05
(SEQ ID NO: 78) 38 Q(AcK)Y QPL DEL DK(0P)T(HChA)YD QFM LQQ G >5K
702.6 (SEQ ID NO: 79) 39 Q(AcK)Y QPL DEL DK(0P)T(HF)YD QFM LQQ G
4894 258.1 (SEQ ID NO: 80) 40 Q(AcK)Y QPL DEL DK(0P)T(ThA)YD QFM
LQQ G >5K 357.8 (SEQ ID NO: 81) 41 Q(AcK)Y QPL DEL
DK(0P)T(Nva)YD QFM LQQ G 1339 23.32 (36) (SEQ ID NO: 82) 42 Q(AcK)Y
QPL DEL DK(0P)T(HL)YD QFM LQQ G 1342 38.15 (42) (SEQ ID NO: 83) 43
Q(AcK)Y QPL DE(AcK) DK(0P)T LYD QFM LQQ G 240 14.515 110 (SEQ ID
NO: 84) (38) 44 Q(AcK)Y QPL DEL DK(0P)T LFD QFM LQQ G 36.9 11.68 58
(31) (SEQ ID NO: 85) 45 Q(AcK)Y Q(HP)L DE(ThA) DK(0P)T L(NO2F)D QFM
24.7 12.7 68 (30) LQQ G (SEQ ID NO: 86) 46 Q(AcK)Y QHPL DEThA
DK(0P)T L(BPA)D QFM LQQ G 82.7 25.21 (32) (SEQ ID NO: 87) 47
Q(AcK)Y Q(HP)L DE(ThA) DK(0P)T L(CO2H)FD QFM 43.3 20.65 (47) LQQ G
(SEQ ID NO: 88) 48 Q(AcK)Y QPL DE(AcK) DK(0P)T L(NO2F)D QFM LQQ
82.4 15.75 64 (35) G (SEQ ID NO: 89) 49 (DCB)Q(AcK)Y QPL DEL
DK(0P)T LYD QFMLQQ G 28.4 12.45 (24) (SEQ ID NO: 90) 50
(DFB)Q(AcK)Y QPL DEL DK(0P)T LYD QFMLQQ G 33 13.56 (23) (SEQ ID NO:
91) 51 (PyC)Q(AcK)Y QPL DEL DK(0P)T LYD QFMLQQ G 14.3 19.38 (18)
(SEQ ID NO: 92) 52 (Amido 2-PEG)Q(AcK)Y QPL DEL DK(0P)T 133.8 18.14
(31) LYDQFMLQQ G (SEQ ID NO: 93) 53 Q(CIBnCarbamate)KY QPL DEL
DK(0P)T LYD QFM 146.9 LQQ G (SEQ ID NO: 94) 54 Q(AcK)Y QPL DEL
D(Dab)(0P)T LYD QFM LQQ G 291.7 22 (SEQ ID NO: 95) 55 Q(AcK)Y QPL
DEL D(Dap)(0P)T LYD QFM LQQ G 598 34.54 (SEQ ID NO: 96) 56 QNY QPL
DEL DK(0P)T L(BPA)D QFM LQQ G 92.2 17.2 (29) (SEQ ID NO: 97) 57 QNY
QPL DEL DK(0P)T L(CF)D QFM LQQ G 110 15.3 (11) (SEQ ID NO: 98) 58
QNY QPL DEL DK(0P)T LYD QFM LQQ G 36.9 12.9 (24) (SEQ ID NO: 99) 59
QRY QPL DEL DK(0P)T LYD QFM LQQ G 176.8 16.1 (SEQ ID NO: 100) 60
QHY QPL DEL DK(0P)T LYD QFM LQQ G 214 14.4 (SEQ ID NO: 101) 61
Q(Nick)Y QPL DEL DK(0P)T LYD QFM LQQ G 83.9 (SEQ ID NO: 102) 62
Q(AcK)Y QPL DE(AcK) DK(0P)T L(CF)D QFM LQQ G 499 41.3 (SEQ ID NO:
103) 63 Q(AcK)Y QPL DE(AcK) DK(0P)T LFD QFM LQQ G (SEQ ID NO:
104)
[0983] IC50 values for exemplary Ang-2 binding peptide compounds as
determined by reverse competitive ELISA are presented in Table 8.
IC50 values are provided for the targeting peptide plus linker (as
shown in FIG. 3) and the targeting peptide linked to an antibody
(P) via the linker of FIG. 3, unless otherwise specified. In Table
8, compounds are conjugated to humanized aldolase antibody h38c2
and the linker structures shown in FIG. 3 (OP).
TABLE-US-00027 TABLE 8 Ang-2 Ang-2 P T1/2 Compound Sequence IC50 nM
Hours 21 QKY QPL DEL DKT LYD QFM LQQ G (SEQ ID NO: 21) 36.39 22
Q(AcK)Y QPL DEL DKT LYD QFM LQQ G (SEQ ID NO: 22) 14.41 27 QKY QPL
DEL DK(0P)T LYD QFM LQQ G (SEQ ID NO: 68) 0.59 28 QNY QPL DEL
DK(0P)T LYD QFM LQQ G (SEQ ID NO: 69) 0.15 29 Q(AcK)Y QPL DEL
DK(0P)T LYD QFM LQQ G (SEQ ID NO: 70) 0.41 30 Q(AcK)Y QPL DEK(0P)
D(AcK)T LYD QFM LQQ G 1.27 72 (SEQ ID NO: 71) 32 Q(AcK)Y QPL DEL
D(AcK)T K(0P)YD QFM LQQ G >100 (SEQ ID NO: 73) 43 Q(AcK)Y QPL
DE(AcK) DK(0P)T LYD QFM LQQ G 0.92 (SEQ ID NO: 84) 44 Q(AcK)Y QPL
DEL DK(0P)T LFD QFM LQQ G (SEQ ID NO: 85) 0.41 48 Q(AcK)Y QPL
DE(AcK) DK(0P)T L(NO2F)D QFM LQQ G 0.33 (SEQ ID NO: 89) 54 Q(AcK)Y
QPL DEL D(Dab)(0P)T LYD QFM LQQ G 0.09 (SEQ ID NO: 95) 55 Q(AcK)Y
QPL DEL D(Dap)(0P)T LYD QFM LQQ G 1.61 (SEQ ID NO: 96) 64
K(0P)(AcK)Y QPL DEL D(AcK)T LYD QFM LQQ G 1.92 24 (SEQ ID NO: 105)
65 QK(0P)Y QPL DEL D(AcK)T LYD QFM LQQ G (SEQ ID NO: 106) 0.31 12
66 Q(AcK)K(0P) QPL DEL D(AcK)T LYD QFM LQQ G 0.23 17 (SEQ ID NO:
107) 67 Q(AcK)Y K(0P)PL DEL D(AcK)T LYD QFM LQQ G N.I. (SEQ ID NO:
108) 68 Q(AcK)Y QK(0P)L DEL D(AcK)T LYD QFM LQQ G 44.21 (SEQ ID NO:
109) 69 Q(AcK)Y QPK(0P) DEL D(AcK)T LYD QFM LQQ G N.I (SEQ ID NO:
110) 70 Q(AcK)Y QPL K(0P)EL D(AcK)T LYD QFM LQQ G N.I. (SEQ ID NO:
111) 71 Q(AcK)Y QPL DK(0P)L D(AcK)T LYD QFM LQQ G O.288 35 (SEQ ID
NO: 112) 72 Q(AcK)Y QPL DEL K(0P)(AcK)T LYD QFM LQQ G N.I (SEQ ID
NO: 113) 73 Q(AcK)Y QPL DEL D(AcK)K(0P) LYD QFM LQQ G 0.11 56 (SEQ
ID NO: 114) 74 Q(AcK)Y QPL DEL D(AcK)T LK(0P)D QFM LQQ G 32.82 (SEQ
ID NO: 115) 75 Q(AcK)Y QPL DEL D(AcK)T LYK(0P) QFM LQQ G 0.19 65
(SEQ ID NO: 116) 76 Q(AcK)Y QPL DEL D(AcK)T LYD K(0P)FM LQQ G 0.27
94 (SEQ ID NO: 117) 77 Q(AcK)Y QPL DEL D(AcK)T LYD QK(0P)M LQQ G
19.53 (SEQ ID NO: 118) 78 Q(AcK)Y QPL DEL D(AcK)T LYD QFK(0P) LQQ G
0.74 72 (SEQ ID NO: 119) 79 Q(AcK)Y QPL DEL D(AcK)T LYD QFM K(0P)QQ
G 0.077 65 (SEQ ID NO: 120) 80 Q(AcK)Y QPL DEL D(AcK)T LYD QFM
LK(0P)Q G 0.11 35 (SEQ ID NO: 121) 81 Q(AcK)Y QPL DEL D(AcK)T LYD
QFM LQK(0P) G 0.27 27 (SEQ ID NO: 122) 82 Q(AcK)Y QPL DEL D(AcK)T
LYD QFM LQQ K(0P) 0.21 20 (SEQ ID NO: 123)
Truncation Study
[0984] Recombinant human Ang-2 was coated on microtiter plates at
0.5 .mu.g/mL in 100 .mu.L, overnight at 2-8.degree. C. Plates were
washed between all steps and all subsequent incubations occurred at
room temperature. Plates were blocked with 250 .mu.L per well of
SuperBlock for 1-3 hours. Test compounds were pre-mixed with
compound 43 linked to linker (as shown in FIGS. 3) and h38c2 at 10
ng/mL. Peptide mixtures in 100 .mu.L were then added at the listed
final concentrations, the plates sealed, and incubated for 1-2
hours. Bound compound 43 was detected with 100 .mu.L of a 1:20,000
dilution of HRP-labeled anti-human IgG reagent for 1-2 hours.
Finally, 100 .mu.L TMB substrate was added for 10 minutes and the
reaction stopped with 100 .mu.L of 2NH.sub.2SO.sub.4 stop solution.
The plate was then read at 450 nm with correction at 650 nm (IC50
values shown in TABLE 9).
TABLE-US-00028 TABLE 9 Ang-2 T Compound Sequence IC50 nM 43(a)
ac-Q(AcK)Y QPL DE(AcK) DK(0P)T LYD QFM LQQ G-am 139 (SEQ ID NO:
191) 83 ac-Q(AcK)Y QPL DE(AcK) DK(0P)T LYD QFM LQQ-am 111 (SEQ ID
NO: 124) 84 ac-Q(AcK)Y QPL DE(AcK) DK(0P)T LYD QFM LQ-am 190 (SEQ
ID NO: 125) 85 ac-Q(AcK)Y QPL DE(AcK) DK(0P)T LYD QFM L-am (SEQ ID
NO: 126) 444 86 ac-Q(AcK)Y QPL DE(AcK) DK(0P)T LYD QFM-am (SEQ ID
NO: 127) 847 87 ac-(AcK)Y QPL DE(AcK) DK(0P)T LYD QFM LQQ G-am
>4000 (SEQ ID NO: 128) 88 ac-Y QPL DE(AcK) DK(0P)T LYD QFM LQQ
G-am (SEQ ID NO: 129) >4000 89 ac-QPL DE(AcK) DK(0P)T LYD QFM
LQQ G-am (SEQ ID NO: 130) >4000 90 ac-PL DE(AcK) DK(0P)T LYD QFM
LQQ G-am (SEQ ID NO: 131) >4000
Xenograft Studies
[0985] Colo205 cells were cultured with 10% FBS RPMI medium and
3.times.10.sup.6 cells in 0.1 ml Hank's balanced salt solution
(HBSS) were injected subcutaneously into the upper right flank of
nude mice. After 7-9 days, animals were randomized into appropriate
number of groups with average tumor size of 200-300 mm.sup.3. Mice
were then treated with the requisite amount of compounds of the
invention and tumor volumes were measured twice a week. Animals
were terminated once their average tumor volume reached 2000
mm.sup.3. Upon termination, tumors were weighed and saved for
further histological studies. Treatment efficacy was evaluated by
measuring the difference in the tumor volumes of treated versus
control groups. Results are reported as % T/C, where % T/C was
calculated as: % T/C=(Vt-V0)/(Ct-00).times.100, where, V0 and Vt
were the average tumor volumes of treated groups at the beginning
and termination of the group. C0 and Ct were the average tumor
volumes of the control group at the beginning and termination of
the group (Table 10).
TABLE-US-00029 TABLE 10 % TC % TC % TC 10 mg/kg 3 mg/kg 1 mg/kg
Compound Sequence 1x/wk 1x/wk 1x/wk 27 QKY QPL DEL DK(0P)T LYD QFM
LQQ G (SEQ ID NO: 68) 52 28 QNY QPL DEL DK(0P)T LYD QFM LQQ G (SEQ
ID NO: 69) 38 34 29 Q(AcK)Y QPL DEL DK(0P)T LYD QFM LQQ G 60 28 9
(SEQ ID NO: 70) 43 Q(AcK)Y QPL DE(AcK) DK(0P)T LYD QFM LQQ G 53 27
34 (SEQ ID NO: 84) 44 Q(AcK)Y QPL DEL DK(0P)T LFD QFM LQQ G 40 22
(SEQ ID NO: 85) 45 Q(AcK)Y Q(HP)L DE(ThA) DK(0P)T L(NO2F)D QFM LQQ
G 47 (SEQ ID NO: 86) 48 Q(AcK)Y QPL DE(AcK) DK(0P)T L(NO2F)D QFM
LQQ G 45 (SEQ ID NO: 89) 58 QNY QPL DEL DK(0P)T LYD QFM LQQ G (SEQ
ID NO: 99) 51 92 PL DEL DK(0P)T LYD QFM LQQ G (SEQ ID NO: 192)
60
Example 29
Synthesis of
##STR00265##
[0986] is provided in FIG. 28.
[0987] The invention thus has been disclosed broadly and
illustrated in reference to representative embodiments described
above. Those skilled in the art will recognize that various
modifications can be made to the present invention without
departing from the spirit and scope thereof. All publications,
patent applications, and issued patents, are herein incorporated by
reference to the same extent as if each individual publication,
patent application or issued patent were specifically and
individually indicated to be incorporated by reference in its
entirety. Definitions that are contained in text incorporated by
reference are excluded to the extent that they contradict
definitions in this disclosure.
[0988] It is appreciated that certain features of the invention,
which are, for clarity, described in the context of separate
embodiments, may also be provided in combination in a single
embodiment. Conversely, various features of the invention which
are, for brevity, described in the context of a single embodiment,
may also be provided separately or in any suitable
sub-combination.
[0989] It is specifically contemplated that any limitation
discussed with respect to one embodiment of the invention may apply
to any other embodiment of the invention. Furthermore, any
composition of the invention may be used in any method of the
invention, and any method of the invention may be used to produce
or to utilize any composition of the invention. In particular, any
aspect of the invention described in the claims, alone or in
combination owth one or more additional claims and/or aspects of
the description, is to be understood as being combinable with other
aspects of the invention set out elsewhere in the claims and/or
description.
[0990] The use of the term "or" in the claims is used to mean
"and/or" unless explicitly indicated to refer to alternatives only
or the alternative are mutually exclusive, although the disclosure
supports a definition that refers to only alternatives and
"and/or."
[0991] As used herein the specification, "a" or "an" may mean one
or more, unless clearly indicated otherwise. As used herein in the
claim (s), when used in conjunction with the word "comprising, "the
words "a" or "an" may mean one or more than one. As used herein
"another" may mean at least a second or more.
[0992] The words "comprises/comprising" and the words
"having/including" when used herein with reference to the present
invention are used to specify the presence of stated features,
integers, steps or components but does not preclude the presence or
addition of one or more other features, integers, steps, components
or groups thereof.
Sequence CWU 1
1
193168DNAHomo sapiensPrimer for generating humanized Vk 1gagctccaga
tgacccagtc tccatcctcc ctgtctgcat ctgtaggtga ccgcgtcacc 60atcacttg
68269DNAHomo sapiensPrimer for generating humanized Vk 2attcagatat
gggctgccat aagtgtgcag gaggctctga ctggagcggc aagtgatggt 60gacgcggtc
69366DNAHomo sapiensPrimer for generating humanized Vk 3tatggcagcc
catatctgaa ttggtatctc cagaaaccag gccagtctcc taagctcctg 60atctat
66465DNAHomo sapiensPrimer for generating humanized Vk 4ctgaaacgtg
atgggacacc actgaaacga ttggacactt tatagatcag gagcttagga 60gactg
65590DNAHomo sapiensPrimer for generating humanized Vk 5agtggtgtcc
catcacgttt cagtggcagt ggttctggca cagatttcac tctcaccatc 60agcagtctgc
aacctgaaga ttttgcagtg 90687DNAHomo sapiensPrimer for generating
humanized Vk 6gatctccacc ttggtccctc cgccgaaagt ataagggagg
tgggtgccct gactacagaa 60gtacactgca aaatcttcag gttgcag 87779DNAHomo
sapiensPrimer for generating humanized VH 7gaggtgcagc tggtggagtc
tggcggtggc ttggtacagc ctggcggttc cctgcgcctc 60tcctgtgcag cctctggct
79881DNAHomo sapiensPrimer for generating humanized VH 8ctccaggccc
ttctctggag actggcggac ccagctcatc caatagttgc taaaggtgaa 60gccagaggct
gcacaggaga g 81981DNAHomo sapiensPrimer for generating humanized VH
9tctccagaga agggcctgga gtgggtctca gagattcgtc tgcgcagtga caactacgcc
60acgcactatg cagagtctgt c 811073DNAHomo sapiensPrimer for
generating humanized VH 10cagatacagc gtgttcttgg aattgtcacg
ggagatggtg aagcggccct tgacagactc 60tgcatagtgc gtg 731176DNAHomo
sapiensPrimer for generating humanized VH 11caattccaag aacacgctgt
atctgcaaat gaacagcctg cgcgccgagg acacgggcat 60ttattactgt aaaacg
761279DNAHomo sapiensPrimer for generating humanized VH
12tgaggagacg gtgaccaggg tgccctggcc ccagtagctg aaactgtaga agtacgtttt
60acagtaataa atgcccgtg 791348DNAHomo sapiensPrimer for generating
humanized VH 13gaggaggagg aggaggagct cactccgagg tgcagctggt ggagtctg
481472DNAHomo sapiensPrimer for generating humanized VL
14gaggaggagg aggagaagct tgttgctctg gatctctggt gcctacgggg agctccagat
60gacccagtct cc 7215336DNAMus musculus 15gacgttgtga tgacccagac
tccactctcc ctgcctgtcc gtcttggaga tcaagcctcc 60atctcttgca gatctagtca
gagccttcta cacacttatg gaagccccta tttaaattgg 120tacctgcaga
agccaggcca gtcgccaaag ctcctgatct acaaagtttc caaccgcttt
180tctggggtcc cagacaggtt cagtggcagt ggatcaggga cagatttcac
actcaggatc 240agcagagtgg aggctgagga tctgggagtt tatttctgct
ctcaaggtac acatcttccg 300tacacgttcg gaggggggac caaactggaa atcaaa
33616351DNAMus musculus 16gaggtgaaac tggtggagtc tggaggaggc
ttggtgcaac ctggaggaac catgaaactc 60tcctgtgaaa tttctggatt aactttcaga
aattattgga tgtcttgggt ccgccagtct 120ccagagaagg ggcttgagtg
ggttgctgaa attagattga gatctgataa ttatgcaaca 180cattatgcgg
agtctgtgaa agggaagttc accatctcaa gagatgattc caaaagtcgt
240ctctacctgc aaatgaacag cttaagaact gaagacactg gaatttatta
ctgtaaaacc 300tatttttact cattttctta ctggggccaa gggactctgg
tcactgtctc t 35117287DNAHomo sapiens 17gacatccaga tgacccagtc
tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca
gagcattagc agctatttaa attggtatca gcagaaacca 120gggaaagccc
ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca
180aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag
tctgcaacct 240gaagattttg caacttacta ctgtcaacag agttacagta cccctcc
2871838DNAHomo sapiens 18gctcactttc ggcggaggga ccaaggtgga gatcaaac
3819296DNAHomo sapiens 19gaggtgcagc tgttggagtc tgggggaggc
ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag cctctggatt cacctttagc
agctatgcca tgagctgggt ccgccaggct 120ccagggaagg ggctggagtg
ggtctcagct attagtggta gtggtggtag cacatactac 180gcagactccg
tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat
240ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaaaga
2962048DNAHomo sapiens 20actactttga ctactggggc caaggaaccc
tggtcaccgt ctcctcag 482122PRTHomo sapiensAnti-angiogenic peptide
21Gln Lys Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 2222PRTArtificialAnti-angiogenic
peptide 22Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20 2322PRTHomo
sapiensAnti-angiogenic peptide 23Gln Asn Tyr Gln Pro Leu Asp Glu
Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly
20 2422PRTArtificialAnti-angiogenic peptide 24Gln Xaa Tyr Gln Pro
Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 2522PRTHomo sapiensAnti-angiogenic peptide 25Gln Asn
Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 2622PRTHomo sapiensAnti-angiogenic
peptide 26Gln Lys Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20 2722PRTHomo
sapiensAnti-angiogenic peptide 27Gln Lys Tyr Gln Pro Leu Asp Glu
Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly
20 2822PRTArtificialAnti-angiogenic peptide 28Gln Asn Tyr Gln Pro
Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 2922PRTArtificialAnti-angiogenic peptide 29Gln Xaa
Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 3022PRTArtificialAnti-angiogenic peptide
30Gln Xaa Tyr Gln Pro Leu Asp Glu Lys Asp Xaa Thr Leu Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 3122PRTArtificialAnti-angiogenic
peptide 31Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Glu Thr Lys Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
3222PRTArtificialAnti-angiogenic peptide 32Gln Xaa Tyr Gln Pro Leu
Asp Glu Leu Asp Xaa Thr Lys Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 3322PRTArtificialAnti-angiogenic peptide 33Gln Xaa Tyr
Gln Pro Leu Asp Glu Lys Asp Glu Thr Leu Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 3422PRTArtificialAnti-angiogenic peptide
34Gln Xaa Tyr Gln Xaa Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 3522PRTArtificialAnti-angiogenic
peptide 35Gln Xaa Tyr Gln Xaa Leu Asp Glu Leu Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
3622PRTArtificialAnti-angiogenic peptide 36Gln Xaa Tyr Gln Pro Leu
Asp Glu Leu Asp Lys Thr Ile Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 3722PRTArtificialAnti-angiogenic peptide 37Gln Xaa Tyr
Gln Pro Leu Asp Glu Ile Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 3822PRTArtificialAnti-angiogenic peptide
38Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Xaa Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 3922PRTArtificialAnti-angiogenic
peptide 39Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Xaa Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
4022PRTArtificialAnti-angiogenic peptide 40Gln Xaa Tyr Gln Pro Leu
Asp Glu Leu Asp Lys Thr Xaa Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 4122PRTArtificialAnti-angiogenic peptide 41Gln Xaa Tyr
Gln Pro Leu Asp Glu Leu Asp Lys Thr Xaa Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 4222PRTArtificialAnti-angiogenic peptide
42Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Xaa Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 4322PRTArtificialAnti-angiogenic
peptide 43Gln Xaa Tyr Gln Pro Leu Asp Glu Xaa Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
4422PRTArtificialAnti-angiogenic peptide 44Gln Xaa Tyr Gln Pro Leu
Asp Glu Leu Asp Lys Thr Leu Phe Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 4522PRTArtificialAnti-angiogenic peptide 45Gln Xaa Tyr
Gln Xaa Leu Asp Glu Xaa Asp Lys Thr Leu Xaa Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 4622PRTArtificialAnti-angiogenic peptide
46Gln Xaa Tyr Gln Xaa Leu Asp Glu Xaa Asp Lys Thr Leu Xaa Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 4722PRTArtificialAnti-angiogenic
peptide 47Gln Xaa Tyr Gln Xaa Leu Asp Glu Xaa Asp Lys Thr Leu Xaa
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
4822PRTArtificialAnti-angiogenic peptide 48Gln Xaa Tyr Gln Pro Leu
Asp Glu Xaa Asp Lys Thr Leu Xaa Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 4922PRTArtificialAnti-angiogenic peptide 49Gln Xaa Tyr
Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 5022PRTArtificialAnti-angiogenic peptide
50Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 5122PRTArtificialAnti-angiogenic
peptide 51Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
5222PRTArtificialAnti-angiogenic peptide 52Gln Xaa Tyr Gln Pro Leu
Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 5322PRTArtificialAnti-angiogenic peptide 53Gln Xaa Tyr
Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 5422PRTArtificialAnti-angiogenic peptide
54Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 5522PRTArtificialAnti-angiogenic
peptide 55Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
5622PRTArtificialAnti-angiogenic peptide 56Gln Asn Tyr Gln Pro Leu
Asp Glu Leu Asp Lys Thr Leu Xaa Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 5722PRTArtificialAnti-angiogenic peptide 57Gln Asn Tyr
Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Xaa Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 5822PRTArtificialAnti-angiogenic peptide
58Gln Asn Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 5922PRTArtificialAnti-angiogenic
peptide 59Gln Arg Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
6022PRTArtificialAnti-angiogenic peptide 60Gln His Tyr Gln Pro Leu
Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 6122PRTArtificialAnti-angiogenic peptide 61Gln Xaa Tyr
Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 6222PRTArtificialAnti-angiogenic peptide
62Gln Xaa Tyr Gln Pro Leu Asp Glu Xaa Asp Lys Thr Leu Xaa Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 6322PRTArtificialAnti-angiogenic
peptide 63Gln Xaa Tyr Gln Pro Leu Asp Glu Xaa Asp Lys Thr Leu Phe
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
6422PRTArtificialAnti-angiogenic peptide 64Gln Xaa Tyr Gln Pro Leu
Asp Glu Leu Asp Lys Thr Leu Xaa Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 6522PRTArtificialAnti-angiogenic peptide 65Gln Xaa Tyr
Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 6622PRTArtificialAnti-angiogenic peptide
66Gln Asn Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 6722PRTArtificialAnti-angiogenic
peptide 67Xaa Lys Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
6822PRTArtificialAnti-angiogenic peptide 68Gln Lys Tyr Gln Pro Leu
Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 6922PRTArtificialAnti-angiogenic peptide 69Gln Asn Tyr
Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 7022PRTArtificialAnti-angiogenic peptide
70Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 7122PRTArtificialAnti-angiogenic
peptide 71Gln Xaa Tyr Gln Pro Leu Asp Glu Xaa Asp Xaa Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
7222PRTArtificialAnti-angiogenic peptide 72Gln Xaa Tyr Gln Pro Leu
Asp Glu Leu Asp Glu Thr Xaa Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 7322PRTArtificialAnti-angiogenic peptide 73Gln Xaa Tyr
Gln Pro Leu Asp Glu Leu Asp Xaa Thr Xaa Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 7422PRTArtificialAnti-angiogenic peptide
74Gln Xaa Tyr Gln Pro Leu Asp Glu Xaa Asp Glu Thr Leu Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 7522PRTArtificialAnti-angiogenic
peptide 75Gln Xaa Tyr Gln Xaa Leu Asp Glu Leu Asp Xaa Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
7622PRTArtificialAnti-angiogenic peptide 76Gln Xaa Tyr Gln Xaa Leu
Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 7722PRTArtificialAnti-angiogenic peptide 77Gln Xaa Tyr
Gln Pro Leu Asp Glu Leu Asp Xaa Thr Ile Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 7822PRTArtificialAnti-angiogenic peptide
78Gln Xaa Tyr Gln Pro Leu Asp Glu Ile Asp Xaa Thr Leu Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 7922PRTArtificialAnti-angiogenic
peptide 79Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Xaa Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
8022PRTArtificialAnti-angiogenic peptide 80Gln Xaa Tyr Gln Pro Leu
Asp Glu Leu Asp Xaa Thr Xaa Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 8122PRTArtificialAnti-angiogenic peptide 81Gln Xaa Tyr
Gln Pro Leu Asp Glu Leu Asp Xaa Thr Xaa Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 8222PRTArtificialAnti-angiogenic peptide
82Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Xaa Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 8322PRTArtificialAnti-angiogenic
peptide 83Gln Xaa Tyr Gln Pro
Leu Asp Glu Leu Asp Xaa Thr Xaa Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 8422PRTArtificialAnti-angiogenic peptide 84Gln Xaa
Tyr Gln Pro Leu Asp Glu Xaa Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 8522PRTArtificialAnti-angiogenic peptide
85Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Phe Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 8622PRTArtificialAnti-angiogenic
peptide 86Gln Xaa Tyr Gln Xaa Leu Asp Glu Xaa Asp Xaa Thr Leu Xaa
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
8722PRTArtificialAnti-angiogenic peptide 87Gln Xaa Tyr Gln Xaa Leu
Asp Glu Xaa Asp Xaa Thr Leu Xaa Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 8822PRTArtificialAnti-angiogenic peptide 88Gln Xaa Tyr
Gln Xaa Leu Asp Glu Xaa Asp Xaa Thr Leu Xaa Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 8922PRTArtificialAnti-angiogenic peptide
89Gln Xaa Tyr Gln Pro Leu Asp Glu Xaa Asp Xaa Thr Leu Xaa Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 9022PRTArtificialAnti-angiogenic
peptide 90Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
9122PRTArtificialAnti-angiogenic peptide 91Gln Xaa Tyr Gln Pro Leu
Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 9222PRTArtificialAnti-angiogenic peptide 92Gln Xaa Tyr
Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 9322PRTArtificialAnti-angiogenic peptide
93Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 9422PRTArtificialAnti-angiogenic
peptide 94Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
9522PRTArtificialAnti-angiogenic peptide 95Gln Xaa Tyr Gln Pro Leu
Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 9622PRTArtificialAnti-angiogenic peptide 96Gln Xaa Tyr
Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 9722PRTArtificialAnti-angiogenic peptide
97Gln Asn Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Xaa Asp Gln 1
5 10 15 Phe Met Leu Gln Gln Gly 20 9822PRTArtificialAnti-angiogenic
peptide 98Gln Asn Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Xaa
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
9922PRTArtificialAnti-angiogenic peptide 99Gln Asn Tyr Gln Pro Leu
Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 10022PRTArtificialAnti-angiogenic peptide 100Gln Arg Tyr
Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe
Met Leu Gln Gln Gly 20 10122PRTArtificialAnti-angiogenic peptide
101Gln His Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln
1 5 10 15 Phe Met Leu Gln Gln Gly 20
10222PRTArtificialAnti-angiogenic peptide 102Gln Xaa Tyr Gln Pro
Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 10322PRTArtificialAnti-angiogenic peptide 103Gln Xaa
Tyr Gln Pro Leu Asp Glu Xaa Asp Xaa Thr Leu Xaa Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 10422PRTArtificialAnti-angiogenic
peptide 104Gln Xaa Tyr Gln Pro Leu Asp Glu Xaa Asp Xaa Thr Leu Phe
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
10522PRTArtificialAnti-angiogenic peptide 105Lys Xaa Tyr Gln Pro
Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 10622PRTArtificialAnti-angiogenic peptide 106Gln Xaa
Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 10722PRTArtificialAnti-angiogenic
peptide 107Gln Xaa Xaa Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
10822PRTArtificialAnti-angiogenic peptide 108Gln Xaa Tyr Xaa Pro
Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 10922PRTArtificialAnti-angiogenic peptide 109Gln Xaa
Tyr Gln Xaa Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 11022PRTArtificialAnti-angiogenic
peptide 110Gln Xaa Tyr Gln Pro Xaa Asp Glu Leu Asp Xaa Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
11122PRTArtificialAnti-angiogenic peptide 111Gln Xaa Tyr Gln Pro
Leu Xaa Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 11222PRTArtificialAnti-angiogenic peptide 112Gln Xaa
Tyr Gln Pro Leu Asp Xaa Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 11322PRTArtificialAnti-angiogenic
peptide 113Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Xaa Xaa Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
11422PRTArtificialAnti-angiogenic peptide 114Gln Xaa Tyr Gln Pro
Leu Asp Glu Leu Asp Xaa Xaa Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 11522PRTArtificialAnti-angiogenic peptide 115Gln Xaa
Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Xaa Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 11622PRTArtificialAnti-angiogenic
peptide 116Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr
Xaa Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
11722PRTArtificialAnti-angiogenic peptide 117Gln Xaa Tyr Gln Pro
Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Xaa 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 11822PRTArtificialAnti-angiogenic peptide 118Gln Xaa
Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15
Xaa Met Leu Gln Gln Gly 20 11922PRTArtificialAnti-angiogenic
peptide 119Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Xaa Leu Gln Gln Gly 20
12022PRTArtificialAnti-angiogenic peptide 120Gln Xaa Tyr Gln Pro
Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Xaa
Gln Gln Gly 20 12122PRTArtificialAnti-angiogenic peptide 121Gln Xaa
Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Xaa Gln Gly 20 12222PRTArtificialAnti-angiogenic
peptide 122Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Xaa Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Xaa Gly 20
12322PRTArtificialAnti-angiogenic peptide 123Gln Xaa Tyr Gln Pro
Leu Asp Glu Leu Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Xaa 20 12421PRTArtificialAnti-angiogenic peptide 124Gln Xaa
Tyr Gln Pro Leu Asp Glu Xaa Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln 20 12520PRTArtificialAnti-angiogenic peptide
125Gln Xaa Tyr Gln Pro Leu Asp Glu Xaa Asp Xaa Thr Leu Tyr Asp Gln
1 5 10 15 Phe Met Leu Gln 20 12619PRTArtificialAnti-angiogenic
peptide 126Gln Xaa Tyr Gln Pro Leu Asp Glu Xaa Asp Xaa Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu 12718PRTArtificialAnti-angiogenic
peptide 127Gln Xaa Tyr Gln Pro Leu Asp Glu Xaa Asp Xaa Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met 12821PRTArtificialAnti-angiogenic peptide
128Xaa Tyr Gln Pro Leu Asp Glu Xaa Asp Xaa Thr Leu Tyr Asp Gln Phe
1 5 10 15 Met Leu Gln Gln Gly 20 12920PRTArtificialAnti-angiogenic
peptide 129Tyr Gln Pro Leu Asp Glu Xaa Asp Xaa Thr Leu Tyr Asp Gln
Phe Met 1 5 10 15 Leu Gln Gln Gly 20
13019PRTArtificialAnti-angiogenic peptide 130Gln Pro Leu Asp Glu
Xaa Asp Xaa Thr Leu Tyr Asp Gln Phe Met Leu 1 5 10 15 Gln Gln Gly
13118PRTArtificialAnti-angiogenic peptide 131Pro Leu Asp Glu Xaa
Asp Xaa Thr Leu Tyr Asp Gln Phe Met Leu Gln 1 5 10 15 Gln Gly
13222PRTArtificialAnti-angiogenic peptide 132Gln Xaa Tyr Gln Pro
Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 13322PRTArtificialAnti-angiogenic peptide 133Gln Xaa
Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Xaa Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 13422PRTArtificialAnti-angiogenic
peptide 134Gln Xaa Tyr Gln Pro Leu Asp Glu Lys Asp Xaa Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
13522PRTArtificialAnti-angiogenic peptide 135Gln Xaa Tyr Gln Pro
Leu Asp Glu Xaa Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 13622PRTArtificialAnti-angiogenic peptide 136Gln Xaa
Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 13722PRTArtificialAnti-angiogenic
peptide 137Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Xaa
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
13822PRTArtificialAnti-angiogenic peptide 138Gln Xaa Tyr Gln Pro
Leu Asp Glu Lys Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 13922PRTArtificialAnti-angiogenic peptide 139Gln Xaa
Tyr Gln Pro Leu Asp Glu Xaa Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 14022PRTArtificialAnti-angiogenic
peptide 140Gln Lys Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
14122PRTArtificialAnti-angiogenic peptide 141Gln Lys Tyr Gln Pro
Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 14222PRTArtificialAnti-angiogenic peptide 142Gln Lys
Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 14322PRTArtificialAnti-angiogenic
peptide 143Gln Lys Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
14422PRTArtificialAnti-angiogenic peptide 144Gln Lys Tyr Gln Pro
Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 14522PRTArtificialAnti-angiogenic peptide 145Gln Lys
Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 14622PRTArtificialAnti-angiogenic
peptide 146Gln Lys Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
14722PRTArtificialAnti-angiogenic peptide 147Gln Lys Tyr Gln Pro
Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 14822PRTArtificialAnti-angiogenic peptide 148Gln Lys
Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 14922PRTArtificialAnti-angiogenic
peptide 149Gln Lys Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
15022PRTArtificialAnti-angiogenic peptide 150Gln Lys Tyr Gln Pro
Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 15122PRTArtificialAnti-angiogenic peptide 151Gln Xaa
Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 15222PRTArtificialAnti-angiogenic
peptide 152Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
15322PRTArtificialAnti-angiogenic peptide 153Gln Xaa Tyr Gln Pro
Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 15422PRTArtificialAnti-angiogenic peptide 154Gln Xaa
Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 15522PRTArtificialAnti-angiogenic
peptide 155Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
15622PRTArtificialAnti-angiogenic peptide 156Gln Xaa Tyr Gln Pro
Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 15722PRTArtificialAnti-angiogenic peptide 157Gln Xaa
Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 15822PRTArtificialAnti-angiogenic
peptide 158Gln Xaa Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr
Asp Gln 1 5 10 15 Phe Met Leu Gln Gln Gly 20
15922PRTArtificialAnti-angiogenic peptide 159Gln Xaa Tyr Gln Pro
Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu
Gln Gln Gly 20 16022PRTArtificialAnti-angiogenic peptide 160Gln Xaa
Tyr Gln Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gln 1 5 10 15
Phe Met Leu Gln Gln Gly 20 16122PRTArtificialAnti-angiogenic
peptide 161Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa Xaa Xaa Xaa Xaa Xaa Xaa
Xaa Xaa 1 5 10 15 Xaa Xaa Xaa Xaa Xaa Xaa 20
16222PRTArtificialAnti-angiogenic peptide 162Xaa Xaa Xaa Xaa Xaa
Xaa Asp Glu Xaa Asp Xaa Xaa Xaa Xaa Asp Xaa 1 5 10 15 Xaa Xaa Xaa
Xaa Xaa Xaa 20 16322PRTArtificialAnti-angiogenic peptide 163Xaa Lys
Xaa Xaa Xaa Xaa Asp Glu Xaa Asp Xaa Xaa Xaa Xaa Asp Xaa 1 5 10 15
Xaa Xaa Xaa Xaa Xaa Xaa 20 16422PRTArtificialAnti-angiogenic
peptide 164Xaa Asn Xaa Xaa Xaa Xaa Asp Glu Xaa Asp Xaa Xaa Xaa Xaa
Asp Xaa 1 5 10 15 Xaa Xaa Xaa Xaa Xaa Xaa 20
16522PRTArtificialAnti-angiogenic peptide 165Xaa Xaa Xaa Xaa Xaa
Xaa Asp Glu Xaa Asp Lys Xaa Xaa Xaa Asp Xaa 1 5 10 15 Xaa Xaa Xaa
Xaa Xaa Xaa 20 16622PRTArtificialAnti-angiogenic peptide 166Gln Xaa
Xaa Gln Xaa Leu Asp Glu Xaa Asp Xaa Thr Xaa Xaa Asp Gln 1 5 10 15
Xaa Met Leu Gln Gln Gly 20 16722PRTArtificialAnti-angiogenic
peptide 167Gln Xaa Xaa Gln Xaa Leu Asp Glu Xaa Asp Lys Thr Xaa Xaa
Asp Gln 1 5
10 15 Xaa Met Leu Gln Gln Gly 20 16822PRTArtificialAnti-angiogenic
peptide 168Gln Lys Xaa Gln Xaa Leu Asp Glu Xaa Asp Lys Thr Xaa Xaa
Asp Gln 1 5 10 15 Xaa Met Leu Gln Gln Gly 20
16922PRTArtificialAnti-angiogenic peptide 169Gln Asn Xaa Gln Xaa
Leu Asp Glu Xaa Asp Lys Thr Xaa Xaa Asp Gln 1 5 10 15 Xaa Met Leu
Gln Gln Gly 20 17022PRTArtificialAnti-angiogenic peptide 170Xaa Xaa
Xaa Xaa Xaa Xaa Asp Glu Xaa Asp Xaa Xaa Leu Xaa Asp Xaa 1 5 10 15
Xaa Xaa Xaa Xaa Xaa Xaa 20 17122PRTArtificialAnti-angiogenic
peptide 171Xaa Lys Xaa Xaa Xaa Xaa Asp Glu Xaa Asp Xaa Xaa Leu Xaa
Asp Xaa 1 5 10 15 Xaa Xaa Xaa Xaa Xaa Xaa 20
17222PRTArtificialAnti-angiogenic peptide 172Xaa Asn Xaa Xaa Xaa
Xaa Asp Glu Xaa Asp Xaa Xaa Leu Xaa Asp Xaa 1 5 10 15 Xaa Xaa Xaa
Xaa Xaa Xaa 20 17322PRTArtificialAnti-angiogenic peptide 173Xaa Xaa
Xaa Xaa Xaa Xaa Asp Glu Xaa Asp Lys Xaa Leu Xaa Asp Xaa 1 5 10 15
Xaa Xaa Xaa Xaa Xaa Xaa 20 17422PRTArtificialAnti-angiogenic
peptide 174Gln Xaa Xaa Gln Xaa Leu Asp Glu Xaa Asp Xaa Thr Leu Xaa
Asp Gln 1 5 10 15 Xaa Met Leu Gln Gln Gly 20
17522PRTArtificialAnti-angiogenic peptide 175Gln Xaa Xaa Gln Xaa
Leu Asp Glu Xaa Asp Lys Thr Leu Xaa Asp Gln 1 5 10 15 Xaa Met Leu
Gln Gln Gly 20 17622PRTArtificialAnti-angiogenic peptide 176Gln Lys
Xaa Gln Xaa Leu Asp Glu Xaa Asp Lys Thr Leu Xaa Asp Gln 1 5 10 15
Xaa Met Leu Gln Gln Gly 20 17722PRTArtificialAnti-angiogenic
peptide 177Gln Asn Xaa Gln Xaa Leu Asp Glu Xaa Asp Lys Thr Leu Xaa
Asp Gln 1 5 10 15 Xaa Met Leu Gln Gln Gly 20
17822PRTArtificialAnti-angiogenic peptide 178Gln Xaa Tyr Gln Xaa
Leu Asp Glu Xaa Asp Xaa Thr Xaa Xaa Xaa Xaa 1 5 10 15 Phe Xaa Xaa
Gln Gln Gly 20 17954DNAArtificialPrimer for generating humanized Vk
179gaggaggagg aggagggccc aggcggccga gctccagatg acccagtctc tcca
5418048DNAArtificialPrimer for generating humanized Vk
180gacagatggt gcagccacag ttcgtttgat ctccaccttg gtccctcc
4818148DNAArtificialPrimer for generating humanized VH
181gctgcccaac cagccatggc cgaggtgcag ctggtggagt ctggggga
4818248DNAArtificialPrimer for generating humanized VH
182gaccgatggg cccttggtgg aggctgagga gacggtgacc agggtgcc
4818322DNAArtificialSequencing primer 183aagacagcta tcgcgattgc ag
2218422DNAArtificialSequencing primer 184ctattgccta cggcagccgc tg
2218524DNAArtificialPrimer for generating humanized VH
185gcccccttat tagcgtttgc catc 2418620DNAArtificialPrimer for
generating humanized VL 186gccatggctg gttgggcagc 2018748DNAHomo
sapiens 187actactttga ctactggggc cagggaaccc tggtcaccgt ctcctcag
4818848DNAHomo sapiens 188gctactttga ctactggggc caagggaccc
tggtcaccgt ctcctcag 48189219PRTArtificialHumanized 38c2 light chain
189Glu Leu Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ser Ser Gln Ser Leu Leu
His Thr 20 25 30 Tyr Gly Ser Pro Tyr Leu Asn Trp Tyr Leu Gln Lys
Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn
Arg Phe Ser Gly Val Pro 50 55 60 Ser Arg Phe Ser Gly Ser Gly Ser
Gly Thr Asp Phe Thr Leu Thr Ile 65 70 75 80 Ser Ser Leu Gln Pro Glu
Asp Phe Ala Val Tyr Phe Cys Ser Gln Gly 85 90 95 Thr His Leu Pro
Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 110 Arg Thr
Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 115 120 125
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 130
135 140 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
Gln 145 150 155 160 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp
Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu
Ser Lys Ala Asp Tyr Glu 180 185 190 Lys His Lys Val Tyr Ala Cys Glu
Val Thr His Gln Gly Leu Ser Ser 195 200 205 Pro Val Thr Lys Ser Phe
Asn Arg Gly Glu Cys 210 215 190448PRTArtificialHumanized 38c2 heavy
chain 190Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro
Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Ser Asn Tyr 20 25 30 Trp Met Ser Trp Val Arg Gln Ser Pro Glu
Lys Gly Leu Glu Trp Val 35 40 45 Ser Glu Ile Arg Leu Arg Ser Asp
Asn Tyr Ala Thr His Tyr Ala Glu 50 55 60 Ser Val Lys Gly Arg Phe
Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln
Met Asn Ser Leu Arg Ala Glu Asp Thr Gly Ile Tyr 85 90 95 Tyr Cys
Lys Thr Tyr Phe Tyr Ser Phe Ser Tyr Trp Gly Gln Gly Thr 100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115
120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr
Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val
Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235
240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln
Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360
365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser
Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445
19122PRTArtificialAnti-angiogenic peptid 191Gln Xaa Tyr Gln Pro Leu
Asp Glu Xaa Asp Xaa Thr Leu Tyr Asp Gln 1 5 10 15 Phe Met Leu Gln
Gln Gly 20 19218PRTArtificialAnti-angiogenic peptide 192Pro Leu Asp
Glu Leu Asp Xaa Thr Leu Tyr Asp Gln Phe Met Leu Gln 1 5 10 15 Gln
Gly 19322PRTArtificialAnti-angiogenic peptide 193Gln Xaa Tyr Gln
Xaa Leu Asp Xaa Xaa Asp Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 15 Phe Xaa
Xaa Gln Gln Gly 20
* * * * *
References