Search

EP-4739713-A1 - ANTI-TROP2 ANTIBODIES

EP4739713A1EP 4739713 A1EP4739713 A1EP 4739713A1EP-4739713-A1

Abstract

Antibodies and antigen-binding fragments thereof specific to Trophoblast cell surface antigen 2 (Trop2). Also provided are anti-Trop2 antibody-drug conjugates (ADCs), Trop2-specific chimeric antigen receptor, and multispecific antigen binding proteins, such as bispecific T cell engagers (BiTEs), which bind to Trop2 and other targets. Further provided are compositions comprising the anti-Trop2 antibodies and antigen-binding fragments thereof, a method of diagnosis and cancer treatment employing the anti-Trop2 antibodies and antigen-binding fragments thereof.

Inventors

  • WANG, CHENG-I
  • WANG, BEI
  • NGOH, Zi Xian Eve
  • MOHD SALLEH, Siti Nazihah
  • YEAP, Yee Chin, Yvonne

Assignees

  • Agency for Science, Technology and Research

Dates

Publication Date
20260513
Application Date
20240702

Claims (20)

  1. 1. An anti-Trop2 antibody or antigen-binding fragment thereof, comprising the complementarity determining region (CDR) sequences selected from the group comprising: (i) a CDRH1 sequence of GGSISSGGYY (SEQ ID NO: 1), a CDRH2 sequence of IYYSGST (SEQ ID NO: 5), a CDRH3 sequence of AREEGIAAAAFDI (SEQ ID NO: 9), a CDRL1 sequence of QSVGSF (SEQ ID NO: 13), a CDRL2 sequence of GAS (SEQ ID NO: 17), and a CDRL3 sequence of QQSDSSPFT (SEQ ID NO: 19); (ii) a CDRH1 sequence of GYTFTSYG (SEQ ID NO: 2), a CDRH2 sequence of ISAYNGNT (SEQ ID NO: 6), a CDRH3 sequence of ARKYSGFDY (SEQ ID NO: 10) a CDRL1 sequence of QSLLHSNGYNY (SEQ ID NO: 14) a CDRL2 sequence of LGS (SEQ ID NO: 18); and a CDRL3 sequence of MQNLQTPWT (SEQ ID NO: 20) (iii) a CDRH1 sequence of GFTFSSYS (SEQ ID NO: 3), a CDRH2 sequence of ISSSSSYI (SEQ ID NO: 7), a CDRH3 sequence of ARDYYDSSGYPYYYYGMDV (SEQ ID NO: 11), a CDRL1 sequence of QSVSSSY (SEQ ID NO: 15), a CDRL2 sequence of GAS (SEQ ID NO: 17), and a CDRL3 sequence of HQSGSSLRT (SEQ ID NO: 21); and (iv) a CDRH1 sequence of GFTFSSYW (SEQ ID NO: 4), a CDRH2 sequence of IKQDGSEK (SEQ ID NO: 8), a CDRH3 sequence of ARDFVDWSATPFDY (SEQ ID NO: 12), a CDRL1 sequence of RSLLHSNGYNY (SEQ ID NO: 16), a CDRL2 sequence of LGS (SEQ ID NO: 18), and a CDRL3 sequence of MQALQIPKT (SEQ ID NO: 22).
  2. 2. The antibody or antigen-binding fragment thereof according to claim 1 , wherein the antibody or antigen-binding fragment thereof is a germline configured counterpart thereof.
  3. 3. The antibody or antigen-binding fragment thereof according to any one of the preceding claims, comprising a VH domain sequence selected from the group comprising: QVQLQESGPGLVKPSQTLSLTCAVSGGSISSGGYYWSWIRQPPGKGLEWIGY IYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREEGIAAA AFDIWGQGTM (SEQ ID NO: 23), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWI SAYNGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARKYSGF DYWGQGTL (SEQ ID NO: 24), EVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWI SAYNGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARKYSGF DYWGQGTL (SEQ ID NO: 25), and EVQLVQSGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSSI SSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARDYYDSS GYPYYYYGMDVWGQGTT (SEQ ID NO: 26), EVQLVQSGGGLVQPGGSLRLSCAASGFTFSSYWMSWVRQAPGKGLEWVAN IKQDGSEKYYVDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARDFVD WSATPFDYWGQGTL (SEQ ID NO: 27), or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto.
  4. 4. The antibody or antigen-binding fragment thereof according to any one of the preceding claims, comprising a VL domain sequence selected from the group comprising: DIQMTQSPSSLSASVGDRVTITCRASQSVGSFLNWYQQKPGKAPKLLIYGASS LQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSDSSPFTFGGGTK (SEQ ID NO: 28), EIVLTQSPSSLSASVGDRVTITCRASQSVGSFLNWYQQKPGKAPQVLIFGASN LESGVPSRFSGRGSGSEFTLTINSLQPEDFATYYCQQSDSSPFTFGGGTK (SEQ ID NO: 29), DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLI YLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQNLQTPWTFG QGTK (SEQ ID NO: 30), EIVLTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIY LGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGIYYCMQNLQTPWTFGQG TK (SEQ ID NO: 31), ETTLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGA SSRATGIPDRFSGSGSGTDFNLTISRLEPEDFAVYYCHQSGSSLRTFGQGTT (SEQ ID NO: 32), and DVVMTQSPLSLPVTPGEPASISCRSSRSLLHSNGYNYLDWYVQKPGQSPQLLI YLGSYRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQIPKTFGQ GTK (SEQ ID NO: 33), or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto.
  5. 5. The antibody or antigen-binding fragment thereof according to any one of the preceding claims, comprising a VH domain and a VL domain selected from the group comprising: (i) a VH domain sequence of: QVQLQESGPGLVKPSQTLSLTCAVSGGSISSGGYYWSWIRQPPGKGL EWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYY CAREEGIAAAAFDIWGQGTM (SEQ ID NO: 23) or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a VL domain sequence of: DIQMTQSPSSLSASVGDRVTITCRASQSVGSFLNWYQQKPGKAPKLLI YGASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSDSSPF TFGGGTK (SEQ ID NO: 28) or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto; (ii) a VH domain sequence of: QVQLQESGPGLVKPSQTLSLTCAVSGGSISSGGYYWSWIRQPPGKGL EWIGYIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYY CAREEGIAAAAFDIWGQGTM (SEQ ID NO: 23) or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a VL domain sequence of: EIVLTQSPSSLSASVGDRVTITCRASQSVGSFLNWYQQKPGKAPQVLI FGASNLESGVPSRFSGRGSGSEFTLTINSLQPEDFATYYCQQSDSSP FTFGGGTK (SEQ ID NO: 29) or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto; (Hi) a VH domain sequence of: Q VQ LVQSG AEVKKPGASVKVSCKASG YTFTSYG I SWVRQA PGQG LE WMGWISAYNGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTA VYYCARKYSGFDYWGQGTL (SEQ ID NO: 24) or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a VL domain sequence of: DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQS PQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQ NLQTPWTFGQGTK (SEQ ID NO: 30) or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto; (iv) a VH domain sequence of: EVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLE WMGWISAYNGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTA VYYCARKYSGFDYWGQGTL (SEQ ID NO: 25) or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a VL domain sequence of: EIVLTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQS PQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGIYYCMQN LQTPWTFGQGTK (SEQ ID NO: 31) or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto; (v) a VH domain sequence of: EVQLVQSGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLE WVSSISSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYY CARDYYDSSGYPYYYYGMDVWGQGTT (SEQ ID NO: 26) or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a VL domain sequence of: ETTLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRL LIYGASSRATGIPDRFSGSGSGTDFNLTISRLEPEDFAVYYCHQSGSSL RTFGQGTT (SEQ ID NO: 32) or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto; and (vi) a VH domain sequence of: EVQLVQSGGGLVQPGGSLRLSCAASGFTFSSYWMSWVRQAPGKGLE WVANIKQDGSEKYYVDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAV YYCARDFVDWSATPFDYWGQGTL (SEQ ID NO: 27) or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a VL domain sequence of: DVVMTQSPLSLPVTPGEPASISCRSSRSLLHSNGYNYLDWYVQKPGQ SPQLLIYLGSYRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQ ALQIPKTFGQGTK (SEQ ID NO: 33) or a variant thereof at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical thereto.
  6. 6. The antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the antibody is a full-length antibody.
  7. 7. The antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the antibody is an IgG antibody, such as an lgG1 antibody.
  8. 8. The antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the antibody or antigen-binding fragment thereof is a fully human antibody or antigen-binding fragment thereof.
  9. 9. The antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the antibody or antigen-binding fragment thereof is specific to human Trop2.
  10. 10. The antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the antibody or antigen-binding fragment thereof exhibits cross-reactivity with more two or more different species.
  11. 11. The antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the antibody or antigen-binding fragment thereof binds to human Trop2 and also cynomolgus Trop2.
  12. 12. The antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the antibody or antigen-binding fragment thereof is conjugated to a therapeutic agent.
  13. 13. The antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the therapeutic agent is an anti-cancer agent, for example a chemotherapeutic agent.
  14. 14. The antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the therapeutic agent is selected from the group comprising MMAE (vedotin), PNU-159682, DX-8951 (exatecan), PBD (pyrrolobenzodiazepine) and DMDM.
  15. 15. An antibody-drug conjugate (ADC) comprising the antibody or antigen binding fragment thereof according to any one of the preceding claims.
  16. 16. A multi-specific antibody comprising an anti-Trop2 binding domain, wherein the anti-Trop2 binding domain is an antibody or antigen-binding fragment thereof according to any one of the preceding claims.
  17. 17. The multi-specific antibody according to claim 16, wherein the multi-specific antibody is an immune cell engager.
  18. 18. The multi-specific antibody according to claim 17, wherein the immune cell engager binds to an immune marker selected from the group consisting of CD3, NKG2D, CD4, CD8, CD16 and CD64.
  19. 19. The multi-specific antibody according to claims 17 or 18, wherein the immune cell engager is selected from the group comprising a T cell engager, an NK cell engager, a monocyte engager and a macrophage engager.
  20. 20. The multi-specific antibody according to claim 19, wherein the immune engager is a bispecific T cell engager (BiTE), such as an inducible BiTE, a non-inducible BiTE or a constitutive expression BiTE.

Description

ANTI-TROP2 ANTIBODIES TECHNICAL FIELD The present disclosure relates broadly to antibodies and antigen-binding fragments thereof specific to trophoblast cell surface antigen 2 (Trop2). In particular, the present disclosure encompasses the nucleotide and amino-acid sequences of 6 human antibodies, or the antigen-binding portions thereof, that specifically target human Trop2, both in solution and on the surface of cells. Also provided are Trop2 antibodydrug conjugates (ADCs) and multispecific antigen binding proteins, such as bispecific T cell engagers (BiTEs), which bind to Trop2 and also other targets. BACKGROUND In 2020, there was an estimated 19.3 million of new cancers reported worldwide, and the cancer incidence is expected to rise to 28.4 million cases in 2040. Among all adult human cancer cases, approximately 90% are solid tumors. Solid tumors such as lung cancer, liver cancer, stomach cancer and female breast cancer are among the leading causes of cancer-related death. The global high cancer incidence burden, coupled with poor treatment outcomes and survival rates with standard treatment modalities of surgery, chemotherapy and radiotherapy presents a strong need for the development of more effective treatment options for cancer. Antibody-drug conjugates (ADCs) are now amongst the fastest growing drug classes in oncology, as they combine the best features of monoclonal antibodies and small molecule drugs to create a single moiety that is highly specific and cytotoxic. Many ADCs have demonstrated impressive activity against treatment-refractory cancers, resulting in their approval for both hematologic malignancies and solid tumor indications. Trophoblast cell surface antigen 2 (Trop2), also known as tumor-associated calcium signal transducer 2, is a 36-kDa single pass transmembrane protein that is highly expressed in a variety of epithelial cancers. It has been demonstrated that Trop2 is involved in multiple intracellular signalling including MAPK and PI3K/AKT pathways which are associated with proliferation, migration, and invasion of cancer cells. Overexpression of Trop2 correlates with poor prognosis in several types of cancers, including breast cancer and non-small cell lung cancer (NSCLC). These characteristics make Trop2 an attractive target for cancer therapy. The anti-Trop2 ADC “Trodelvy” developed by ImmunoMedics is one of the latest ADCs approved in 2021. It is the first and the only ADC to treat the devastating metastatic triple negative breast cancer (TNBC). It was subsequently approved to treat the metastatic urothelial cancer in the same year. Apart from its success in various clinical studies, “Trodelvy” represents one of the largest transactions in the biotech history: a $21 billion acquisition by Gilead in 2020. AstraZeneca also entered a $1 billion upfront deal with Daiichi Sankyo to codevelop an ADC against Trop2 DS-1062, with a further $5 billion in milestones. Anti-Trop2 ADCs are thus far approved only for TNBC and urothelial cancer, hence there is a need for additional and superior anti-Trop2 ADCs that can treat these indications as well as other cancers. SUMMARY The present inventors have successfully identified four anti-Trop2 antibody clones from their nai ve human Fab phage libraries, two of them were further engineered to their germline configured counterparts. One or more of these antibodies displayed 1) superior binding affinity and specificity to the Trop2 protein, both in solution and expressed on the cell surface; 2) superior cytotoxic potency when used in ADCC; 3) superior growth inhibition potency when used as ADC; 4) strong cytotoxic potency when used as bi-specific T cell engager (BiTE) antibodies; and 5) strong cytotoxicity potency when used in CAR T cell therapy. Thus, in one aspect, there is provided an anti-Trop2 antibody or antigen-binding fragment thereof, comprising the CDR sequences selected from the group comprising: (i) a CDRH1 sequence of GGSISSGGYY (SEQ ID NO: 1), a CDRH2 sequence of IYYSGST (SEQ ID NO: 5), a CDRH3 sequence of AREEGIAAAAFDI (SEQ ID NO: 9), a CDRL1 sequence of QSVGSF (SEQ ID NO: 13), a CDRL2 sequence of GAS (SEQ ID NO: 17), and a CDRL3 sequence of QQSDSSPFT (SEQ ID NO: 19); (ii) a CDRH1 sequence of GYTFTSYG (SEQ ID NO: 2), a CDRH2 sequence of ISAYNGNT (SEQ ID NO: 6), a CDRH3 sequence of ARKYSGFDY (SEQ ID NO: 10) a CDRL1 sequence of QSLLHSNGYNY (SEQ ID NO: 14) a CDRL2 sequence of LGS (SEQ ID NO: 18); and a CDRL3 sequence of MQNLQTPWT (SEQ ID NO: 20) (iii) a CDRH1 sequence of GFTFSSYS (SEQ ID NO: 3), a CDRH2 sequence of ISSSSSYI (SEQ ID NO: 7), a CDRH3 sequence of ARDYYDSSGYPYYYYGMDV (SEQ ID NO: 11), a CDRL1 sequence of QSVSSSY (SEQ ID NO: 15), a CDRL2 sequence of GAS (SEQ ID NO: 17), and a CDRL3 sequence of HQSGSSLRT (SEQ ID NO: 21); and (iv) a CDRH1 sequence of GFTFSSYW (SEQ ID NO: 4), a CDRH2 sequence of IKQDGSEK (SEQ ID NO: 8), a CDRH3 sequence of ARDFVDWSATPFDY (SEQ ID NO: 12), a CDRL1 sequence of RSLLHSNGYNY (SEQ ID NO: 16), a CDR