CN-120173111-B - Anti-platelet GPIB-IX antibody and application thereof

Abstract

The invention discloses an anti-platelet GPIB-IX antibody and application thereof, belonging to the technical field of biological medicine. The invention selects an initial signal egg GPIB-IX of arterial thrombosis as a target point to prepare the humanized antibody of the anti-human GPIB-IX. Specifically, a mouse is immunized by using human platelets as antigens, a murine anti-human GPIB-IX antibody is obtained by screening, an anti-GPIB-IX humanized antibody ATHA is constructed by mutating a human sequence, the heavy chain variable region of the anti-GPIB-IX humanized antibody comprises a VH-CDR3 shown as SEQ ID NO.1, and the light chain variable region comprises a VL-CDR3 shown as SEQ ID NO. 2. The Fab fragment ATHA can effectively inhibit platelet activation mediated by GPIB-IX receptor, inhibit thrombosis in vivo, and has the advantages of no adverse side effects of thrombocytopenia and hemorrhage, high specificity, good safety, and small adverse side effects. At the same time ATHA can remove platelets. Therefore, ATHA-Fab has clinical value as a new anti-thrombus medicine and can be used for treating diseases related to arterial thrombosis. ATHA can remove platelets, and can be used for treating diseases with increased platelet count or removing platelets in vivo.

Inventors

• DAI KESHENG

Assignees

• 苏州大学

Dates

Publication Date

20260512

Application Date

20250213

Claims (9)

1. 1. A monoclonal antibody directed against platelet GPIb-IX, wherein the monoclonal antibody comprises a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID No.3 and a light chain variable region comprising an amino acid sequence as set forth in SEQ ID No. 5.
2. 2. The monoclonal antibody of claim 1, comprising at least one of the following: (1) The monoclonal antibody contains a heavy chain constant region, wherein the heavy chain constant region comprises an amino acid sequence with homology of not less than 90% with SEQ ID NO. 4; (2) The monoclonal antibody contains a light chain constant region, wherein the light chain constant region comprises an amino acid sequence having homology of not less than 90% with SEQ ID NO. 6.
3. 3. The monoclonal antibody of claim 2, wherein the heavy chain constant region comprises an amino acid sequence as set forth in SEQ ID No. 4; The light chain constant region comprises an amino acid sequence as shown in SEQ ID NO. 6.
4. 4. A polynucleotide encoding the monoclonal antibody of any one of claims 1-3.
5. 5. A recombinant expression vector comprising the polynucleotide of claim 4.
6. 6. A recombinant cell comprising the recombinant expression vector of claim 5.
7. 7. A monovalent, divalent or multivalent antibody comprising the monoclonal antibody of any one of claims 1-3.
8. 8. A prophylactic and/or therapeutic agent comprising the monoclonal antibody according to any one of claims 1 to 3, the polynucleotide according to claim 4, the recombinant expression vector according to claim 5, the recombinant cell according to claim 6, or the monovalent, bivalent or multivalent antibody according to claim 7.
9. 9. A monoclonal antibody according to any one of claims 1-3, a polynucleotide according to claim 4, a recombinant expression vector according to claim 5, a recombinant cell according to claim 6, a monovalent, bivalent or multivalent antibody according to claim 7 or a medical use of a prophylactic and/or therapeutic drug according to claim 8, characterized in that the medical use is: (1) Is used for preparing antithrombotic medicines; (2) Is used for preparing medicines for removing blood platelets.

Description

Anti-platelet GPIB-IX antibody and application thereof Technical Field The invention relates to an anti-platelet GPIB-IX antibody and application thereof, belonging to the technical field of biological medicine. Background Arterial thrombosis diseases such as myocardial infarction, coronary heart disease and cerebral infarction seriously threaten human health and life, and coronary heart disease occupies the first ten orders of world population total-cause mortality published by WHO in 2022. Platelets play a critical role in the development and progression of arterial thrombotic disease. As the disease progresses, activated platelets can cause excessive thrombosis, leading to vascular obstruction and impaired organ function. Currently, antiplatelet agents are the primary agents for the prevention and treatment of such diseases, reducing the occurrence of thrombosis and ischemic complications by inhibiting platelet activation. Therefore, the antiplatelet medicine can save the life of the patient, improve the life quality of the patient and has important clinical application and research value. Conventional antiplatelet agents can be classified into TXA2 inhibitors, phosphodiesterase activators, P2Y12 receptor antagonists, ADP receptor antagonists, GPIIb/IIIa receptor antagonists, 5-HT receptor antagonists, PAR1 receptor antagonists, and the like, according to the effect of the agent on the different targets in the platelet activation signal pathway. Although these antiplatelet drugs can reduce the occurrence of thrombotic diseases, side effects of the drugs are also found, for example, 1% of patients may directly induce platelet aggregation due to the combination of the drugs and platelets, namely thrombocytopenia, drug resistance phenomena caused by drug absorption, metabolism and interaction, and most commonly, bleeding risk is increased. Therefore, the ideal antiplatelet medicine should firstly reduce thrombosis and have no bleeding side effect, and secondly have the characteristics of long half life, wide safe and effective dose, small immunogenicity and the like. At present, a plurality of scholars are still working on developing and improving anti-platelet drugs, and hope to obtain novel drugs with better curative effect and better safety along with the continuous deep research of the foundation and the clinic, break the clinical dilemma of anti-thrombosis treatment, and enable thrombotic patients to obtain longer survival time and better life quality. When arterial vascular endothelial cells are damaged or under the action of high shearing force, platelets are combined with von Willebrand factor (von Willebrand Factor, VWF) on the surface of the damaged vascular endothelial through glycoprotein (Glycoprotein, GP) Ib-IX receptors on the surface of the arterial vascular endothelial cells, and the platelets adhere to the surface of the damaged vascular endothelial cells while the platelets are activated, and aggregate with each other to form thrombus to play a hemostatic function. The prior studies indicate that soybean helps to prevent cardiovascular disease in humans, namely because the isoflavones enriched in soybean inhibit GPIB function. And inhibiting GPIB-IX function can also improve brain blood circulation, reduce cerebral ischemia/reperfusion injury and inhibit clinical symptoms of vascular restenosis. Thus, GPIB-IX is an initial and key protein for forming arterial thrombosis by blood platelets, and is a first-choice target for developing a novel medicine for preventing and treating arterial thrombosis. Since the first murine mab drug was approved by the U.S. Food and Drug Administration (FDA) in 1986, nearly hundred mab drugs have been marketed worldwide. The targeted GPIB-IX monoclonal antibody drug is the most potential novel anti-thrombus drug. Many drugs currently under investigation targeting the GPIb-IX signaling pathway, such as AP-1, PP403C, PG-1, 6B4, 24G10, SZ2, h6B4-Fab, AJW200, where SZ2 shows advantages over other drugs both in platelet binding and in platelet aggregation inhibition, but these agents under investigation may lead to acute thrombocytopenia and increased platelet deacetylation side effects. There has been no approved GPIb-IX antibody drug for clinical use. The humanized monoclonal antibody can overcome the human anti-mouse antibody (HAMA) reaction, can effectively reduce the HAMA reaction induced by the mouse antibody in clinical use, has the characteristics of high affinity, high specificity, low immunogenicity and small toxic and side effects, effectively improves the drug effect of the monoclonal antibody drug, and becomes the key point of the development of the current therapeutic antibody drug. To make up for the blank of the GPIB-IX antibody drug market, the side effect of drug-induced thrombocytopenia is solved. We have developed humanized antibodies against GPIb-IX for the prevention and treatment of arterial thrombosis related disorders. Disclosure of Invent