Targeted CD73 antibody and antibody-drug conjugate, and preparation method therefor and uses thereof

Patent metadata

Field	Value
Publication number	US-12024565-B2
Application number	US-201916978995-A
Country	US
Kind code	B2
Filing date	Mar 7, 2019
Priority date	Mar 7, 2018
Publication date	Jul 2, 2024
Grant date	Jul 2, 2024

Abstract

Disclosed by the present invention are a targeted CD73 antibody and an antibody-drug conjugate (ADC), and a preparation method therefor and application thereof. Further disclosed is a method for preparing the described monoclonal antibody and ADC. The monoclonal antibody and the corresponding ADC disclosed by the present invention can be efficiently and highly specifically combined with purified CD73 protein and CD73 on the surfaces of multiple tumor cells to block the catalytic activity of CD73 enzyme, and have high affinity, low immunogenicity and significant anti-tumor effect.

First claim

The invention claimed is:

1. An antibody, which binds to CD73, wherein the antibody comprises a heavy chain and a light chain,
wherein the heavy chain has a
heavy chain variable region comprising the following three complementarity determining regions (CDRs):
CDR1 as shown in SEQ ID NO. 10,
CDR2 as shown in SEQ ID NO. 11, and
CDR3 as shown in SEQ ID NO. 12; and
the light chain has a light chain variable region comprising the following three complementarity determining regions (CDRs):
CDR1′ as shown in SEQ ID NO. 13,
CDR2′ as shown in SEQ ID NO. 14, and
CDR3′ as shown in SEQ ID NO. 15;
or
the heavy chain has a heavy chain variable region comprising the following three complementary determining regions (CDRs):
CDR1 as shown in SEQ ID NO. 1,
CDR2 as shown in SEQ ID NO. 2, and
CDR3 as shown in SEQ ID NO. 3; and
the light chain has a light chain variable region comprising the following three complementarity determining regions (CDRs):
CDR1′ as shown in SEQ ID NO. 4,
CDR2′ as shown in SEQ ID NO. 5, and
CDR3′ as shown in SEQ ID NO. 6;
or
the heavy chain has a heavy chain variable region comprising the following three complementary determining regions (CDRs):
CDR1 as shown in SEQ ID NO. 21,
CDR2 as shown in SEQ ID NO. 22, and
CDR3 as shown in SEQ ID NO. 23; and/or
the light chain has a light chain variable region comprising the following three complementarity determining regions (CDRs):
CDR1′ as shown in SEQ ID NO. 24,
CDR2′ as shown in SEQ ID NO. 25, and
CDR3′ as shown in SEQ ID NO. 26 or
the heavy chain has a heavy chain variable region comprising the following three complementary determining regions or CDRs:
CDR1 as shown in SEQ ID NO. 1,
CDR2 as shown in SEQ ID NO. 2, and
CDR3 as shown in SEQ ID NO. 3; wherein the sixth amino acid of CDR2 is mutated to Q or the seventh amino acid of CDR2 is mutated to A; and
the light chain has a light chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1′ as shown in SEQ ID NO. 4,
CDR2′ as shown in SEQ ID NO. 5, and
CDR3′ as shown in SEQ ID NO. 6;
or
the heavy chain has a heavy chain variable region comprising the following three complementary determining regions or CDRs:
CDR1 as shown in SEQ ID NO. 1,
CDR2 as shown in SEQ ID NO. 2, and
CDR3 as shown in SEQ ID NO. 3; and
the light chain has a light chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1′ as shown in SEQ ID NO. 4,
CDR2′ as shown in SEQ ID NO. 5, and
CDR3′ as shown in SEQ ID NO. 6; wherein the seventh amino acid of CDR2′ is mutated to S;
or
the heavy chain has a heavy chain variable region comprising the following three complementary determining regions or CDRs:
CDR1 as shown in SEQ ID NO. 1,
CDR2 as shown in SEQ ID NO. 2, and
CDR3 as shown in SEQ ID NO. 3; wherein the sixth amino acid of CDR2 is mutated to Q; and
the light chain has a light chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1′ as shown in SEQ ID NO. 4,
CDR2′ as shown in SEQ ID NO. 5, and
CDR3′ as shown in SEQ ID NO. 6; wherein the seventh amino acid of CDR2′ is mutated to S;
or
the heavy chain has a heavy chain variable region comprising the following three complementary determining regions or CDRs:
CDR1 as shown in SEQ ID NO. 21,
CDR2 as shown in SEQ ID NO. 22, and
CDR3 as shown in SEQ ID NO. 23; wherein the sixth amino acid of CDR2 is mutated to Q or the seventh amino acid of CDR2 is mutated to A; and
the light chain has a light chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1′ as shown in SEQ ID NO. 24,
CDR2′ as shown in SEQ ID NO. 25, and

CDR3′ as shown in SEQ ID NO. 26.

2. A recombinant protein which comprises:
(i) the antibody of claim 1 ; and
(ii) an optional tag sequence that assists expression and/or purification.

3. A CAR construct comprising a scFv segment of a monoclonal antibody that specifically binds to CD73, wherein the scFv segment of the monoclonal antibody antigen binding region of the CAR construct is a binding region that specifically binds to CD73, and
the scFv has a heavy chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1 as shown in SEQ ID NO. 10,
CDR2 as shown in SEQ ID NO. 11, and
CDR3 as shown in SEQ ID NO. 12; and
a light chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1′ as shown in SEQ ID NO. 13,
CDR2′ as shown in SEQ ID NO. 14, and
CDR3′ as shown in SEQ ID NO. 15;
or
the scFv has a heavy chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1 as shown in SEQ ID NO. 1,
CDR2 as shown in SEQ ID NO. 2, and
CDR3 as shown in SEQ ID NO. 3; and
a light chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1′ as shown in SEQ ID NO. 4,
CDR2′ as shown in SEQ ID NO. 5, and
CDR3′ as shown in SEQ ID NO. 6;
or
the scFv has a heavy chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1 as shown in SEQ ID NO. 21,
CDR2 as shown in SEQ ID NO. 22, and
CDR3 as shown in SEQ ID NO. 23; and
a light chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1′ as shown in SEQ ID NO. 24,
CDR2′ as shown in SEQ ID NO. 25, and
CDR3′ as shown in SEQ ID NO. 26 or
the heavy chain has a heavy chain variable region comprising the following three complementary determining regions or CDRs:
CDR1 as shown in SEQ ID NO. 1,
CDR2 as shown in SEQ ID NO. 2, and
CDR3 as shown in SEQ ID NO. 3; wherein the sixth amino acid of CDR2 is mutated to Q or the seventh amino acid of CDR2 is mutated to A; and
the light chain has a light chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1′ as shown in SEQ ID NO. 4,
CDR2′ as shown in SEQ ID NO. 5, and
CDR3′ as shown in SEQ ID NO. 6;
or
the heavy chain has a heavy chain variable region comprising the following three complementary determining regions or CDRs:
CDR1 as shown in SEQ ID NO. 1,
CDR2 as shown in SEQ ID NO. 2, and
CDR3 as shown in SEQ ID NO. 3; and
the light chain has a light chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1′ as shown in SEQ ID NO. 4,
CDR2′ as shown in SEQ ID NO. 5, and
CDR3′ as shown in SEQ ID NO. 6; wherein the seventh amino acid of CDR2′ is mutated to S;
or
the heavy chain has a heavy chain variable region comprising the following three complementary determining regions or CDRs:
CDR1 as shown in SEQ ID NO. 1,
CDR2 as shown in SEQ ID NO. 2, and
CDR3 as shown in SEQ ID NO. 3; wherein the sixth amino acid of CDR2 is mutated to Q; and
the light chain has a light chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1′ as shown in SEQ ID NO. 4,
CDR2′ as shown in SEQ ID NO. 5, and
CDR3′ as shown in SEQ ID NO. 6; wherein the seventh amino acid of CDR2′ is mutated to S;
or
the heavy chain has a heavy chain variable region comprising the following three complementary determining regions or CDRs:
CDR1 as shown in SEQ 1D NO. 21,
CDR2 as shown in SEQ ID NO. 22, and
CDR3 as shown in SEQ ID NO. 23; wherein the sixth amino acid of CDR2 is mutated to Q or the seventh amino acid of CDR2 is mutated to A; and
the light chain has a light chain variable region comprising the following three complementarity determining regions or CDRs:
CDR1′ as shown in SEQ ID NO. 24,
CDR2′ as shown in SEQ ID NO. 25, and

CDR3′ as shown in SEQ ID NO. 26.

4. A recombinant immune cell expressing an exogenous CAR c

Assignees

• Univ Fudan
• Bliss Biopharmaceutical Hangzhou Co Ltd

Inventors

• Yu Ke
• Jin Rui
• Liu Liang

Classifications

• A61K47/68031

  the drug being an auristatin · CPC title

• C07K16/40

  against enzymes · CPC title

• A61K2039/51

  comprising whole cells, viruses or DNA/RNA · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K2317/77

  Internalization into the cell · CPC title