Inhibition of angiogenesis by peptide analogs of high...

Details Inhibition of angiogenesis by peptide analogs of high... Inhibition of angiogenesis by peptide analogs of high...

2000-07-07

2001-09-04

Carlson, Karen Cochrane (Department: 1653)

Drug, bio-affecting and body treating compositions

Designated organic active ingredient containing

Peptide containing doai

C514S002600, C514S012200, C514S016700, C435S006120, C435S069800, C435S069100, C435S091500, C604S890100

Reexamination Certificate

active

06284726

ABSTRACT:

FIELD OF THE INVENTION
The invention relates to therapeutic compounds and methods for inhibiting angiogenesis.
BACKGROUND OF THE INVENTION
Angiogenesis
Angiogenesis is the process in which new blood vessels grow into an area which lacks a sufficient blood supply. Angiogenesis commences with the erosion of the basement membrane surrounding endothelial cells and pericytes forming capillary blood vessels. Erosion of the basement membrane is triggered by enzymes released by endothelial cells and leukocytes. The endothelial cells then migrate through the eroded basement membrane when induced by angiogenic stimulants. The migrating cells form a “sprout” off the parent blood vessel. The migrating endothelial cells proliferate, and the sprouts merge to form capillary loops, thus forming a new blood vessel.
Angiogenesis can occur under certain normal conditions in mammals such as in wound healing, in fetal and embryonic development, and in the formation of the corpus luteum, endometrium and placenta. Angiogenesis also occurs in certain disease states such as in tumor formation and expansion, or in the retina of patients with certain ocular disorders. Angiogenesis can also occur in a rheumatoid joint, hastening joint destruction by allowing an influx of leukocytes with subsequent release of inflammatory mediators.
The evidence for the role of angiogenesis in tumor growth was extensively reviewed and present by O'Reilly and Folkman in U.S. Pat. No. 5,639,725, the entire disclosure of which is incorporated herein by reference. It is now generally accepted that the growth of tumors is critically dependent upon this process.
High Molecular Weight Kininogen
High molecular weight kininogen (HK) is a 120 kD glycoprotein containing heavy and light chains, composed of domains 1 through 3, and 5 and 6, respectively. This form of HK is often referred to as “single-chain high molecular weight kininogen”. HK binds with high affinity to endothelial cells, where it is cleaved by plasma kallikrein into heavy and light chains. This form of cleaved HK is known as “two-chain high molecular weight kininogen” (HK
a
). The heavy and light chains are linked by domain 4 in intact HK; domain 4 contains bradykinin, a potent biologically active nonapeptide. Bradykinin is released from HK through cleavage mediated by plasma kallikrein (Kaplan et al.,
Blood
70:1-15, 1987). The heavy and light chains resulting from the elimination of bradykinin remain linked by a disulfide bond between cysteine residues at positions 10 and 596. Conversion of HK to HK
a
is accompanied by a dramatic structural rearrangement. The central globular region of HK is separated after bradykinin liberation and rearranged with cysteine protease inhibitory regions opposite the prekallikrein binding region (Weisel et al.,
J. Biol. Chem.
262:1405, 1987).
The HK light chain consists of the following sequence of HK amino acids 372-626 (SEQ ID NO:1):
SSRIGEIKEETTVSPPHTSMAPAQDEERD SGKEQGHTRRHDWGHEKQRKHNLGHGH KHERDQGHGHQRGHGLGHGHEQQHGLG HGHKFKLDDDLEHQGGHVLDHGHKHKH GHGHGKHKNKGKKNGKHNGWKTEHLAS SSEDSTTPSAQTQEKTEGPTPIPSLAKPG VTVTFSDFQDSDLIATMMPPISPAPIQSD DDWIPDIQTDPNGLSFNPISDFPDTTSPK CPGRPWKSVSEINPTTQMKESYYFDLTD GLS  (SEQ ID NO:1).
The HK light chain consists of HK amino acids serine-372 to threonine 383, forming the C-terminal portion of HK domain 4 remaining after bradykinin liberation; HK amino acids valine 384 to lysine 502, forming the HK domain 5 (D5); and HK amino acids threonine 503 to serine 626, forming the HK domain 6 (D6). D5 is rich in histidines, glycines and lysines and has been postulated to be involved in binding to negatively charged surfaces. D5 serves as a cell binding site on platelets, granulocytes and endothelial cells. For a recent review including a discussion of HK's domain structure, and the functional significance of the various domains (including D5), see Colman and Schmaier,
Blood
90:3819-3843 (1997). HK
a
has been shown to bind to the urokinase receptor (uPAR) on endothelial cells (Colman et al.,
J. Clin. Invest.
100:1481-7 (1997). HK D5 has been shown to participate in cell binding; certain peptides from regions of D5 have been found to inhibit HK binding to endothelial cells (Hasan et al.,
J. Biol. Chem.,
270:19256, 1995). Despite these findings, no role has been suggested for HK in the process of angiogenesis.
SUMMARY OF THE INVENTION
The compounds of the present invention are in the form of peptides which possess anti-angiogenic activity.
In all embodiments, the peptide may optionally comprise an amino-terminal and/or carboxy-terminal protecting group.
Compounds possessing anti-angiogenic activity are derived from HK domain 5 and 6 have the formula
X
1
-(SEQ ID NO:2)-X
2
  (I)
wherein
X
1
is from zero to 25 amino acids;
X
2
is from zero to 60 amino acids; and
SEQ ID NO:2 is the amino acid sequence corresponding to HK light chain His(441)-His(455): HGLGHGHEQQ HGLGH.
According to one embodiment of the invention, X
1
is from zero to 12 amino acids, more preferably from zero to 6 amino acids; and X
2
is from zero to 45 amino acids, more preferably from zero to 32 amino acids, more preferably from zero to 22 amino acids, most preferably from zero to six amino acids.
In certain preferred compounds,
X
1
is
(i) zero amino acids, or
(ii) the segment KHNLGHGHKHE RDQGHGHQRG (SEQ ID NO:3), or N-terminal truncation fragment thereof containing at least one amino acid, and
X
2
is
(i) zero amino acids, or
(ii) the segment GHKFKLDDDLEHQG GHVLDHGHKHKHGHGHGKHKNKGK KNGKHNGWK (SEQ ID NO:4), or C-terminal truncation fragment thereof containing at least one amino acid.
SEQ ID NO:3 and NO:4 correspond to HK light chain amino acids Lys(420)-Gly(440) and Gly(456)-Lys(502), respectively.
According to a further preferred embodiment of the invention, X
1
has a substantial amino acid homology to SEQ ID NO:3 and X
2
has a substantial amino acid homology to SEQ ID NO:4.
Exemplary and preferred compounds of formula I include:
(a) GHGLGHGHEQQHGLGH (SEQ ID NO:9), corresponding to HK light chain amino acids Gly(440)-His(455);
(b) KHNLGHGHKHERDQGHGHQRG HGLGHGHEQQHGLGHGHKFKL DDDLEHQGGHVLD (SEQ ID NO:5), corresponding to HK light chain amino acids Lys(420)-Asp(474);
(c) KHNLGHGHKHERDQGHGHQRG HGLGHGHEQQHGLGHGHKFKL DDDLEHQGGHVLDHGHKHKHG HGHGKHKNKGKKNGKHNGWK (SEQ ID NO:6), corresponding to HK light chain amino acids Lys(420)-Lys(502);
(d) HGLGHGHEQQHGLGHGHKFKL DDDLEHQGGHVLDHGHKHKHG HGHGKHKNKGKKNGKHNGWK (SEQ ID NO:7), corresponding to HK light chain amino acids His(441)-Lys(502); and
(e) KHNLGHGHKHERDQGHGHQRG HGLGHGHEQQHGLGHGHKFKL DDDLEHQGGHVLDHGHKHKHG HGHGKHKNKGKKNGKHNGWKT EHLASSSEDS (SEQ ID NO:10), corresponding to HK light chain amino acids Lys(420)-Ser(513).
According to a related aspect of the invention, the anti-angiogenic compound has the amino acid sequence of HK light chain amino acids His(475)-His(485), that is, SEQ ID NO:11: HGHKHKHG HGH.
According to another related aspect of the invention, the anti-angiogenic compound has the amino acid sequence of HK light chain amino acids His(441)-Ser(626), that is, SEQ ID NO:8:
HGLGHGHEQQHGLGHGHKFKL DDDLEHQGGHVLDHGHKHKHG HGHGKHKNKGKKNGKHNGWKT EHLASSSEDSTTPSAQTQEKTE GPTPIPSLAKPGVTVTFSDFQD SDLIATMMPPISPAPIQSDDDW IPDIQTDPNGLSFNPISDFPDTT SPKCPGRPWKSVSEINPTTQMK ESYYFDLTDGLS  (SEQ ID NO:8).
According to another related aspect of the invention, the anti-angiogenic compound has the amino acid sequence of HK light chain amino acids Glu(448)-Ser(626), that is, SEQ ID NO:12:
EQQHGLGHGHKFKLDDDLEHQ GGHVLDHGHKHKHGHGHGKHK NKGKKNGKHNGWKTEHLASSS EDSTTPSAQTQEKTEGPTPIPS LAKPGVTVTFSDFQDSDLIATM MPPISPAPIQSDDDWIPDIQTDP NGLSFNPISDFPDTTSPKCPGR PWKSVSEINPTTQMKESYYFDL TDGLS  (SEQ ID NO:12).
SEQ ID NOS:8 and 12 begin in domain 5 and extend through the entirety of domain 6.
The invention also encompasses a method of inhibiting endothelial cell proliferation comprising contacting endothelial cells with a compound of formula I, or with the compound of SEQ ID NO:8, 11 or 12.
The invention also encompasses a method of inhibiting migration of endothelial cells to vitronectin

Affiliated with

Also associated with

Rating

Inhibition of angiogenesis by peptide analogs of high... does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Inhibition of angiogenesis by peptide analogs of high..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Inhibition of angiogenesis by peptide analogs of high... will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2509896