Inhibition of Plasma Kallikrein by a Highly Specific Active Site Blocking Antibody

被引：102

作者：

Kenniston, Jon A. ^{[1
]}

Faucette, Ryan R. ^{[1
]}

Martik, Diana ^{[1
]}

Comeau, Stephen R. ^{[1
]}

Lindberg, Allison P. ^{[1
]}

Kopacz, Kris J. ^{[1
]}

Conley, Gregory P. ^{[1
]}

Chen, Jie ^{[1
]}

Viswanathan, Malini ^{[1
]}

Kastrapeli, Niksa ^{[1
]}

Cosic, Janja ^{[1
]}

Mason, Shauna ^{[1
]}

DiLeo, Mike ^{[1
]}

Abendroth, Jan ^{[2
]}

Kuzmic, Petr ^{[3
]}

Ladner, Robert C. ^{[1
]}

Edwards, Thomas E. ^{[2
]}

TenHoor, Christopher ^{[1
]}

Adelman, Burt A. ^{[1
]}

Nixon, Andrew E. ^{[1
]}

Sexton, Daniel J. ^{[1
]}

机构：

[1] Dyax Corp, Burlington, MA 01803 USA

[2] Beryllium, Bainbridge Isl, WA 98110 USA

[3] Biokin, Watertown, MA 02472 USA

来源：

JOURNAL OF BIOLOGICAL CHEMISTRY | 2014年 / 289卷 / 34期

关键词：

Antibody Engineering; Enzyme Inhibitor; Inflammation; Kallikrein; Protease; MOLECULAR-WEIGHT KININOGEN; INDUCED PAW EDEMA; HEREDITARY ANGIOEDEMA; FACTOR-XII; CONTACT SYSTEM; C1-ESTERASE INHIBITOR; BLOOD-COAGULATION; C1; INHIBITOR; IN-VITRO; ACTIVATION;

D O I：

10.1074/jbc.M114.569061

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Background: Unregulated plasma kallikrein proteolytic activity can result from C1-inhibitor deficiency, causing excessive and potentially fatal edema. Results: The antibody DX-2930 potently and specifically inhibits plasma kallikrein and exhibits a long plasma half-life. Conclusion: An antibody protease inhibitor can lead to potent and specific bioactivity. Significance: DX-2930 could be an effective therapeutic for the prophylactic inhibition of plasma kallikrein-mediated diseases. Plasma kallikrein (pKal) proteolytically cleaves high molecular weight kininogen to generate the potent vasodilator and the pro-inflammatory peptide, bradykinin. pKal activity is tightly regulated in healthy individuals by the serpin C1-inhibitor, but individuals with hereditary angioedema (HAE) are deficient in C1-inhibitor and consequently exhibit excessive bradykinin generation that in turn causes debilitating and potentially fatal swelling attacks. To develop a potential therapeutic agent for HAE and other pKal-mediated disorders, we used phage display to discover a fully human IgG1 monoclonal antibody (DX-2930) against pKal. In vitro experiments demonstrated that DX-2930 potently inhibits active pKal (K-i = 0.120 +/- 0.005 nm) but does not target either the zymogen (prekallikrein) or any other serine protease tested. These findings are supported by a 2.1- resolution crystal structure of pKal complexed to a DX-2930 Fab construct, which establishes that the pKal active site is fully occluded by the antibody. DX-2930 injected subcutaneously into cynomolgus monkeys exhibited a long half-life (t(1/2) approximate to 12.5 days) and blocked high molecular weight kininogen proteolysis in activated plasma in a dose- and time-dependent manner. Furthermore, subcutaneous DX-2930 reduced carrageenan-induced paw edema in rats. A potent and long acting inhibitor of pKal activity could be an effective treatment option for pKal-mediated diseases, such as HAE.

引用

页码：23596 / 23608

页数：13

共 79 条

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