Fully human anti-RANKL antibody

An antibody and whole-person technology, applied in the direction of antibodies, anti-inflammatory agents, anti-tumor drugs, etc., can solve problems such as increased difficulty, difficult production control, difficult quality control, etc.

Active Publication Date: 2015-11-25
SHANGHAI JMT BIO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition to the high risk of immunosuppression, it is difficult to control the production of denosumab due to the complexity of its IgG2 subtype and free cysteine ​​structure, which makes it more difficult to develop the production process and GMP production. Quality Control

Method used

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  • Fully human anti-RANKL antibody
  • Fully human anti-RANKL antibody
  • Fully human anti-RANKL antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1: Design and expression of BA05-1 and BA05-2 antibody sequences

[0076] The following is the sequence of the heavy chain variable region of AMG162:

[0077] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSGITGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDPGTTVIMSWFDPWGQGTLVTVSS (SEQ ID NO: 9)

[0078] The following is the heavy chain constant region sequence of AMG162:

[0079] ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:10)

[0080] The original heavy chain IgG2 constant region sequence was replaced with IgG4 constant region sequence to form the BA05-1 heavy chain sequence:

[0081] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSGITGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAE...

Embodiment 2

[0094] Example 2: Expression and preparation of BA05-1 and BA05-2 antibodies

[0095] The full-length DNA sequence of the heavy chain of BA05-1 is:

[0096]GAGGTGCAGCTCCTGGAGAGCGGCGGAGGCCTGGTGCAGCCCGGAGGAAGCCTGCGGCTCTCCTGCGCCGCTAGCGGATTCACATTCTCCAGCTACGCTATGAGCTGGGTCAGGCAGGCTCCTGGCAAGGGACTCGAGTGGGTGAGCGGCATCACCGGATCCGGCGGATCCACATACTATGCCGATTCCGTCAAGGGAAGGTTCACAATCTCCCGGGACAACAGCAAGAACACCCTCTACCTCCAGATGAACAGCCTGCGGGCCGAGGACACAGCCGTCTACTATTGCGCCAAAGACCCCGGAACCACCGTGATCATGAGCTGGTTCGATCCCTGGGGACAGGGAACCCTCGTGACAGTGTCCAGCGCTAGCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCATGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCCAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGG...

Embodiment 3

[0106] Example 3: Detection of the binding of BA05-1 and BA05-2 antibodies to RANKL by flow cytometry

[0107] Binding of anti-RANKL antibodies BA05-1 and BA05-2 to RANKL (full-length RANKL) transfected with RANKL on the surface of 293 cells (Invitrogen) was detected by flow cytometry. 293-RANKL transfected cells were incubated with different concentrations of anti-RANKL antibodies BA05-1, BA05-2, and AMG162 at 4°C for 1 h. The cells were washed with PBS, and anti-human IgG (H+L)--PE (1:200) / anti-human IgG-R-PE (1:1000) antibodies were added to incubate at 4°C for 30 min. After washing with PBS, the cells were resuspended in 1 ml of PBS and analyzed by flow cytometry (BDFACScan). Such as figure 1 As shown, the binding ability of BA05-1 to RANKL is equivalent to that of AMG162, while the binding ability of BA05-2 to RANKL is slightly better than that of AMG162.

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Abstract

The present invention relates to fully humanized antibodies, specifically to a fully human anti-RANKL antibody, a nucleic acid molecule encoding the antibody, and a composition containing the antibody. The present invention further relates to uses of the antibody in preparation of drugs for prevention and treatment of bone loss related diseases, especially osteoporosis, arthritis osteolysis damage or tumor bone metastasis and other diseases. According to the present invention, the antibody is specifically bound with human RANKL and blocks the activity of the human RANKL while the potential immunesuppression of the human RANKL on human body is reduced, the antibody has good stability, and the defect of the non-uniformity (heterogeneity) of the IgG2 antibody production on the batch quality identification is avoided.

Description

technical field [0001] The present invention relates to a fully humanized antibody, specifically, the present invention relates to a fully human anti-RANKL antibody, a nucleic acid molecule encoding the antibody and a composition comprising the antibody, and the present invention also relates to the preparation of the antibody for the prevention and treatment of bone Use in medicine for diseases such as porosis, osteolytic destruction of arthritis, or tumor bone metastasis. Background technique [0002] Bone formation and bone remodeling are coordinated by bone resorbing osteoclasts and bone forming osteoblasts, which are responsible for bone metabolism and bone remodeling in human development and growth. Bone is continuously resorbed and formed at specific locations in the skeleton, and it is estimated that about 10% of the total bone in the human body is rebuilt each year. Osteoclasts derived from monocyte / macrophage precursors can resorb bone after activation, and eventu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28C12N15/13C12N15/70C12N1/21A61K39/395A61K48/00A61P29/00A61P37/00A61P19/10A61P19/08A61P19/02A61P35/00G01N33/68
Inventor 刘劼
Owner SHANGHAI JMT BIO INC
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