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CN-113861292-B - Anti-CD 40 antibody or antigen binding fragment and application thereof

CN113861292BCN 113861292 BCN113861292 BCN 113861292BCN-113861292-B

Abstract

The invention provides an anti-CD 40 antibody or antigen binding fragment, which specifically binds to CD40, and can activate antigen presenting cells, stimulate the antigen presenting cells to release cytokines, induce apoptosis of tumor cells and inhibit proliferation of the tumor cells. Also provided is the use of the above antibody or antigen binding fragment in the manufacture of a medicament for treating or ameliorating a CD 40-associated disorder, or in the manufacture of a kit for diagnosing a CD 40-associated disorder.

Inventors

  • ZHANG HUI
  • HUANG XIANMING
  • YUE RUI
  • CHEN ZHENCHENG
  • WANG ZHIWEI
  • CHEN JUNYOU
  • LI SHENGFENG

Assignees

  • 百奥泰生物制药股份有限公司

Dates

Publication Date
20260512
Application Date
20210629
Priority Date
20200630

Claims (20)

  1. 1. An anti-CD 40 antibody or antigen-binding fragment thereof, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a VL CDR1, a VL CDR2, a VL CDR3, a VH CDR1, a VH CDR2 and a VH CDR3, wherein the amino acid sequence of the VL CDR1 is shown as SEQ ID NO. 1, the amino acid sequence of the VL CDR2 is shown as SEQ ID NO. 2, the amino acid sequence of the VL CDR3 is shown as SEQ ID NO. 3, the amino acid sequence of the VH CDR1 is shown as SEQ ID NO. 4, the amino acid sequence of the VH CDR2 is shown as SEQ ID NO. 48, the amino acid sequence of the VH CDR3 is shown as SEQ ID NO. 79, or The amino acid sequence of the VL CDR1 is shown as SEQ ID NO. 1, the amino acid sequence of the VL CDR2 is shown as SEQ ID NO. 2, the amino acid sequence of the VL CDR3 is shown as SEQ ID NO. 3, the amino acid sequence of the VH CDR1 is shown as SEQ ID NO. 5, the amino acid sequence of the VH CDR2 is shown as SEQ ID NO. 48, the amino acid sequence of the VH CDR3 is shown as SEQ ID NO. 79, or The amino acid sequence of the VL CDR1 is shown as SEQ ID NO. 1, the amino acid sequence of the VL CDR2 is shown as SEQ ID NO. 2, the amino acid sequence of the VL CDR3 is shown as SEQ ID NO. 3, the amino acid sequence of the VH CDR1 is shown as SEQ ID NO. 18, the amino acid sequence of the VH CDR2 is shown as SEQ ID NO. 60, the amino acid sequence of the VH CDR3 is shown as SEQ ID NO. 79, or The amino acid sequence of the VL CDR1 is shown as SEQ ID NO. 1, the amino acid sequence of the VL CDR2 is shown as SEQ ID NO. 2, the amino acid sequence of the VL CDR3 is shown as SEQ ID NO. 3, the amino acid sequence of the VH CDR1 is shown as SEQ ID NO. 6, the amino acid sequence of the VH CDR2 is shown as SEQ ID NO. 49, the amino acid sequence of the VH CDR3 is shown as SEQ ID NO. 79, or The amino acid sequence of the VL CDR1 is shown as SEQ ID NO.1, the amino acid sequence of the VL CDR2 is shown as SEQ ID NO.2, the amino acid sequence of the VL CDR3 is shown as SEQ ID NO.3, the amino acid sequence of the VH CDR1 is shown as SEQ ID NO. 28, the amino acid sequence of the VH CDR2 is shown as SEQ ID NO. 65, the amino acid sequence of the VH CDR3 is shown as SEQ ID NO. 79, or The amino acid sequence of the VL CDR1 is shown as SEQ ID NO. 1, the amino acid sequence of the VL CDR2 is shown as SEQ ID NO. 2, the amino acid sequence of the VL CDR3 is shown as SEQ ID NO. 3, the amino acid sequence of the VH CDR1 is shown as SEQ ID NO. 7, the amino acid sequence of the VH CDR2 is shown as SEQ ID NO. 49, the amino acid sequence of the VH CDR3 is shown as SEQ ID NO. 79, or The amino acid sequence of the VL CDR1 is shown as SEQ ID NO.1, the amino acid sequence of the VL CDR2 is shown as SEQ ID NO.2, the amino acid sequence of the VL CDR3 is shown as SEQ ID NO.3, the amino acid sequence of the VH CDR1 is shown as SEQ ID NO. 29, the amino acid sequence of the VH CDR2 is shown as SEQ ID NO. 66, the amino acid sequence of the VH CDR3 is shown as SEQ ID NO. 79, or The amino acid sequence of the VL CDR1 is shown as SEQ ID NO.1, the amino acid sequence of the VL CDR2 is shown as SEQ ID NO.2, the amino acid sequence of the VL CDR3 is shown as SEQ ID NO.3, the amino acid sequence of the VH CDR1 is shown as SEQ ID NO. 32, the amino acid sequence of the VH CDR2 is shown as SEQ ID NO. 48, the amino acid sequence of the VH CDR3 is shown as SEQ ID NO. 79, or The amino acid sequence of the VL CDR1 is shown as SEQ ID NO.1, the amino acid sequence of the VL CDR2 is shown as SEQ ID NO.2, the amino acid sequence of the VL CDR3 is shown as SEQ ID NO. 3, the amino acid sequence of the VH CDR1 is shown as SEQ ID NO. 8, the amino acid sequence of the VH CDR2 is shown as SEQ ID NO. 50, the amino acid sequence of the VH CDR3 is shown as SEQ ID NO. 79, or The amino acid sequence of the VL CDR1 is shown as SEQ ID NO.1, the amino acid sequence of the VL CDR2 is shown as SEQ ID NO.2, the amino acid sequence of the VL CDR3 is shown as SEQ ID NO.3, the amino acid sequence of the VH CDR1 is shown as SEQ ID NO. 33, the amino acid sequence of the VH CDR2 is shown as SEQ ID NO. 69, the amino acid sequence of the VH CDR3 is shown as SEQ ID NO. 79, or The amino acid sequence of the VL CDR1 is shown as SEQ ID NO.1, the amino acid sequence of the VL CDR2 is shown as SEQ ID NO.2, the amino acid sequence of the VL CDR3 is shown as SEQ ID NO. 3, the amino acid sequence of the VH CDR1 is shown as SEQ ID NO. 9, the amino acid sequence of the VH CDR2 is shown as SEQ ID NO. 48, the amino acid sequence of the VH CDR3 is shown as SEQ ID NO. 79, or The amino acid sequence of the VL CDR1 is shown as SEQ ID NO. 1, the amino acid sequence of the VL CDR2 is shown as SEQ ID NO. 2, the amino acid sequence of the VL CDR3 is shown as SEQ ID NO. 3, the amino acid sequence of the VH CDR1 is shown as SEQ ID NO. 40, the amino acid sequence of the VH CDR2 is shown as SEQ ID NO. 49, and the amino acid sequence of the VH CDR3 is shown as SEQ ID NO. 79.
  2. 2. The anti-CD 40 antibody or antigen-binding fragment thereof of claim 1, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence set forth in any one of SEQ ID NOs 81-86, 96, 106, 107, 110, 111, or 118.
  3. 3. The anti-CD 40 antibody or antigen-binding fragment thereof according to claim 1, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a light chain variable region comprising the amino acid sequence set forth in SEQ ID No. 80.
  4. 4. The anti-CD 40 antibody or antigen-binding fragment thereof according to claim 2, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a light chain variable region comprising the amino acid sequence set forth in SEQ ID No. 80.
  5. 5. The anti-CD 40 antibody or antigen-binding fragment thereof of claim 1, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence as set forth in any one of SEQ ID NOs 81-86, 96, 106, 107, 110, 111 or 118 and a light chain variable region having an amino acid sequence as set forth in SEQ ID NO 80.
  6. 6. The anti-CD 40 antibody or antigen-binding fragment thereof according to claim 2, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence as set forth in any one of SEQ ID NOs 81 to 86, 96, 106, 107, 110, 111 or 118 and a light chain variable region having an amino acid sequence as set forth in SEQ ID NO 80.
  7. 7. The anti-CD 40 antibody or antigen-binding fragment thereof according to claim 3, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence as set forth in any one of SEQ ID NOs 81 to 86, 96, 106, 107, 110, 111 or 118 and a light chain variable region having an amino acid sequence as set forth in SEQ ID NO 80.
  8. 8. The anti-CD 40 antibody or antigen-binding fragment thereof according to claim 4, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence as set forth in any one of SEQ ID NOs 81 to 86, 96, 106, 107, 110, 111 or 118 and a light chain variable region having an amino acid sequence as set forth in SEQ ID NO 80.
  9. 9. The anti-CD 40 antibody or antigen-binding fragment thereof of any one of claims 1-8, wherein the anti-CD 40 antibody or antigen-binding fragment thereof is one of the isoforms IgG, igM, igA, igE or IgD.
  10. 10. The anti-CD 40 antibody or antigen-binding fragment thereof of any one of claims 1-8, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a light chain constant region as set forth in SEQ ID No. 127, and the anti-CD 40 antibody or antigen-binding fragment thereof comprises a heavy chain constant region as set forth in SEQ ID No. 128.
  11. 11. The anti-CD 40 antibody or antigen-binding fragment thereof of claim 9, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a light chain constant region as set forth in SEQ ID No. 127, and wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a heavy chain constant region as set forth in SEQ ID No. 128.
  12. 12. The anti-CD 40 antibody or antigen-binding fragment thereof of any one of claims 1-8, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a heavy chain comprising a heavy chain variable region as set forth in any one of SEQ ID NOs 81-86, 96, 106, 107, 110, 111, or 118 and a heavy chain constant region as set forth in SEQ ID NO 128, and a light chain comprising a light chain variable region as set forth in SEQ ID NO 80 and a light chain constant region as set forth in SEQ ID NO 127.
  13. 13. The anti-CD 40 antibody or antigen-binding fragment thereof of claim 9, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a heavy chain comprising a heavy chain variable region as set forth in any one of SEQ ID NOs 81-86, 96, 106, 107, 110, 111, or 118, and a heavy chain constant region as set forth in SEQ ID NO 128, and a light chain comprising a light chain variable region as set forth in SEQ ID NO 80, and a light chain constant region as set forth in SEQ ID NO 127.
  14. 14. The anti-CD 40 antibody or antigen-binding fragment thereof of claim 10, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a heavy chain comprising a heavy chain variable region as set forth in any one of SEQ ID NOs 81-86, 96, 106, 107, 110, 111, or 118, and a heavy chain constant region as set forth in SEQ ID No. 128, and a light chain comprising a light chain variable region as set forth in SEQ ID No. 80, and a light chain constant region as set forth in SEQ ID No. 127.
  15. 15. The anti-CD 40 antibody or antigen-binding fragment thereof of claim 11, wherein the anti-CD 40 antibody or antigen-binding fragment thereof comprises a heavy chain comprising a heavy chain variable region as set forth in any one of SEQ ID NOs 81-86, 96, 106, 107, 110, 111, or 118, and a heavy chain constant region as set forth in SEQ ID No. 128, and a light chain comprising a light chain variable region as set forth in SEQ ID No. 80, and a light chain constant region as set forth in SEQ ID No. 127.
  16. 16. An anti-CD 40 antibody or antigen-binding fragment thereof, wherein said anti-CD 40 antibody or antigen-binding fragment thereof comprises a light chain having the amino acid sequence shown in SEQ ID No. 129, and said anti-CD 40 antibody or antigen-binding fragment thereof comprises a heavy chain having the amino acid sequence shown in SEQ ID No. 130.
  17. 17. A nucleic acid molecule encoding an anti-CD 40 antibody or antigen-binding fragment thereof according to any one of claims 1 to 16.
  18. 18. A vector comprising the nucleic acid molecule of claim 17.
  19. 19. A host cell comprising the nucleic acid molecule of claim 17.
  20. 20. A kit comprising an anti-CD 40 antibody or antigen-binding fragment thereof according to any one of claims 1 to 16.

Description

Anti-CD 40 antibody or antigen binding fragment and application thereof Technical Field The invention belongs to the field of biotechnology, and particularly relates to an anti-CD 40 antibody or antigen binding fragment and application thereof. Background CD40 receptor and its ligand CD40L (CD 154) belong to the TNFR: TNF family. The CD40-CD40L signaling pathway in B cells has an effect on the production of long-lived plasma cells and memory B cells and their survival. CD40 belongs to a co-stimulatory receptor expressed on antigen presenting cells such as Dendritic Cells (DCs), B cells, monocytes, and some other non-lymphocytes. CD40-CD40L interactions (an important signal) play an important role in regulating the interactions of DC cells with T cells (CD4+ T cells and CD8+ T cells), rescuing the depleted CTL, reversing the tolerance of DC cells. In addition, CD40-CD40L interactions between B cells and T cells affect germinal center (GERMINAL CENTER) reactions, inducing antibody isotype switching. CD40 expressed on T cells interacts with CD154 expressed on APC, which results in maturation of DC cells, increases cytokine production and costimulatory factor expression, and can increase differentiation of effector T cells. CD40 is a line I transmembrane glycoprotein, CD40L is a type II transmembrane glycoprotein, and as with other members of the TNFR superfamily, CD40 forms a trimer. Although CD40 is believed to be the primary binding receptor for CD40L, CD40L may also bind to other receptors of the integrin family, such as αIIbβ3, α5β1 (VLA-5), αMβ2 (Mac-1). The CD40 agonist antibody can activate and stimulate innate immunity, act on adaptive immunity and produce direct cytotoxicity to CD40 expressing malignant tumor cell. CD40 is a promising therapeutic target for tumor immunotherapy. Disclosure of Invention The present invention provides antibodies or antigen binding fragments that specifically bind CD 40. In some embodiments, the invention provides antibodies or antigen binding fragments that specifically bind to human CD40 or monkey CD 40. The anti-CD 40 antibody of the invention can activate and stimulate innate immunity after being specifically combined with CD40, and can generate direct cytotoxicity for nausea tumor cells expressing CD 40. In some embodiments, the invention discloses an antibody or antigen binding fragment that specifically binds CD40 comprising one or more of VH CDR1, VH CDR2 and VH CDR3, wherein VH CDR1 comprises the amino acid sequence X1-X2-X3-X4-X5-X6-X7, X1 is D, F, G, or Y, X2 is I, N, S or T, X3 is F, X4 is A, D, F, G, T, S or Y, X5 is D, G, N, S or T, X6 is A, F, N, S or Y, X7 is A, S or Y, and X1-X2-X3-X4-X5-X6-X7 comprises at least three different amino acids, VH CDR2 comprises the amino acid sequence Z1-Z2-Z3-Z4-Z5-Z6-Z7-Z8-Z9-Z10, Z1 is A, G or S, Z2 is I, Z3 is D, I, K or S, Z4 is F, X4 is A, D, F, G, T, S or Y, X6 is A, F, N, S or Y, X7 is A, S or Y, and X1-X4-X5-X6-X7 comprises at least three different amino acids, VH CDR2 comprises the amino acid sequence Z1-Z2-Z4-Z5-Z6-Z8-Z9-Z10, Z3 is 3875 or Z3 is shown as Y, Z3 is 3835 or Z3 is shown as Y, Y3 is 3-3 or Y, Z3 is 3 or Z9, and at least 3 is a Z9-Z9 or Z9. In some embodiments, the invention discloses an antibody or antigen-binding fragment that specifically binds CD40, comprising one or more of VH CDR1, VH CDR2, and VH CDR3, wherein VH CDR1 comprises the amino acid sequence of X1-X2-X3-X4-X5-X6-X7, X1 is D, F, G, or Y, X2 is I, N, S or T, X3 is F, X4 is A, D, F, G, T, S or Y, X5 is D, G, N, S or T, X6 is A, F, N, S or Y, X7 is A, S or Y, and X1-X2-X3-X4-X5-X6-X7 comprises at least three different amino acids, VH CDR2 comprises the amino acid sequence of SEQ ID NO 70 or 74, and VH CDR3 comprises the amino acid sequence of SEQ ID NO 79. In some embodiments, X1 is F, X4 is D or G, X5 is S or T, and X6 is F, N or Y. In some embodiments, Z1 is G, Z3 is D or S, Z4 is S or W, Z5 is D or G, Z6 is G or T, Z7 is D or G, Z8 is N, S or T, Z9 is a or P, and Z10 is F, H or S. In some embodiments, the VH CDR1 comprises an amino acid sequence as set forth in any one of SEQ ID NOs 4-47. In some embodiments, the VH CDR2 comprises an amino acid sequence as set forth in any one of SEQ ID NOS: 48-78. In some embodiments, the antibody or antigen binding fragment further comprises one or more of a VL CDR1, a VL CDR2, and a VL CDR3, the VL CDR1 comprising the amino acid sequence shown in SEQ ID No. 1, the VL CDR2 comprising the amino acid sequence shown in SEQ ID No.2, and the VL CDR3 comprising the amino acid sequence shown in SEQ ID No. 3. In some embodiments, the antibody or antigen-binding fragment comprises a VH CDR1, a VH CDR2, a VH CDR3, a VL CDR1, a VL CDR2, and a VL CDR3. In some embodiments, the antibody or antigen-binding fragment specifically binds CD40, the antibody or antigen-binding fragment comprises one or more of VL CDR1, VL CDR2, VL CDR3, VH CDR1, VH CDR2 and VH CDR3, the VL CDR1 comprising the amino acid sequence as set fo