title: Simultaneous targeting of CD14 and factor XIa by a fusion protein consisting of an anti-CD14 antibody and the modified second domain of bikunin improves survival in rabbit sepsis models.
abstract: Severe sepsis is a complex, multifactorial, and rapidly progressing disease characterized by excessive inflammation and coagulation following bacterial infection. To simultaneously suppress pro-inflammatory and pro-coagulant responses, we genetically engineered a novel fusion protein (MR1007) consisting of an anti-CD14 antibody and the modified second domain of bikunin, and evaluated the potential of MR1007 as an anti-sepsis agent. Suppressive effects of MR1007 on lipopolysaccharide (LPS)-induced inflammatory responses were assessed using peripheral blood mononuclear cells or endothelial cells. Its inhibitory activity against the coagulation factor XIa was assessed using a purified enzyme and a chromogenic substrate. Anticoagulant activity was assessed using human or rabbit plasma. Anti-inflammatory and anti-coagulant effects and/or survival benefits were evaluated in an endotoxemia model and a cecal ligation and puncture model. MR1007 inhibited LPS-induced cytokine production in peripheral blood mononuclear cells and endothelial cells, inhibited factor XIa, and exhibited anticoagulant activity. In an endotoxemia model, 0.3-3mg/kg MR1007 suppressed pro-inflammatory and pro-coagulant responses in a dose-dependent manner; at a dose of 3mg/kg, the protein improved survival even when administered 8h after the LPS injection. In addition, 10mg/kg MR1007 administered 2h post cecal ligation and puncture improved survival. However, MR1007 administered at doses up to 30mg/kg did not increase ear bleeding time or bacterial counts in the cecal ligation and puncture model. Thus, simultaneous targeting of CD14 and factor XIa improves survival in the rabbit endotoxemia and sepsis models and represents a promising approach for the treatment of severe sepsis.
authors.affiliation: Discovery Research, Mochida Pharmaceutical Co., Ltd., 722 Jimba-aza-Uenohara, Gotemba, Shizuoka 412-8524, Japan. Electronic address: mnakamur@mochida.co.jp.
authors.forename: Masaki
authors.initials: M
authors.lastname: Nakamura
authors.affiliation: Discovery Research, Mochida Pharmaceutical Co., Ltd., 722 Jimba-aza-Uenohara, Gotemba, Shizuoka 412-8524, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Takeuchi
authors.affiliation: Discovery Research, Mochida Pharmaceutical Co., Ltd., 722 Jimba-aza-Uenohara, Gotemba, Shizuoka 412-8524, Japan.
authors.forename: Tetsushi
authors.initials: T
authors.lastname: Kawahara
authors.affiliation: Discovery Research, Mochida Pharmaceutical Co., Ltd., 722 Jimba-aza-Uenohara, Gotemba, Shizuoka 412-8524, Japan.
authors.forename: Jiro
authors.initials: J
authors.lastname: Hirose
authors.affiliation: Discovery Research, Mochida Pharmaceutical Co., Ltd., 722 Jimba-aza-Uenohara, Gotemba, Shizuoka 412-8524, Japan.
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Nakayama
authors.affiliation: Discovery Research, Mochida Pharmaceutical Co., Ltd., 722 Jimba-aza-Uenohara, Gotemba, Shizuoka 412-8524, Japan.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Hosaka
authors.affiliation: Discovery Research, Mochida Pharmaceutical Co., Ltd., 722 Jimba-aza-Uenohara, Gotemba, Shizuoka 412-8524, Japan.
authors.forename: Shoji
authors.initials: S
authors.lastname: Furusako
mesh_terms: D000510:Alpha-Globulins / Q000737:chemistry*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology* / Q000627:therapeutic use; D000925:Anticoagulants / Q000276:immunology / Q000494:pharmacology / Q000627:therapeutic use; D001760:Bleeding Time; D004195:Disease Models, Animal; D019040:E-Selectin / Q000378:metabolism; D019446:Endotoxemia / Q000188:drug therapy* / Q000378:metabolism; D015945:Factor XIa / Q000037:antagonists & inhibitors*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis; D018950:Lipopolysaccharide Receptors / Q000276:immunology*; D058990:Molecular Targeted Therapy; D000072417:Protein Domains; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000494:pharmacology* / Q000627:therapeutic use; D016019:Survival Analysis
chemical_list: D000510:Alpha-Globulins; D000911:Antibodies, Monoclonal; D000925:Anticoagulants; D019040:E-Selectin; D015850:Interleukin-6; D018950:Lipopolysaccharide Receptors; D011993:Recombinant Fusion Proteins; C016001:alpha-1-microglobulin; D015945:Factor XIa
keywords: Bikunin; Heparin sodium salt (PubChem CID: 57199466); Inter-⍺ inhibitor; Lipopolysaccharide; Lipopolysaccharide (PubChem CID: 11970143); Ulinastatin; Urinary trypsin inhibitor