Office Action 1: Non-Final Rejection (2024-09-10)
Rejection types: 102 - Lack of Novelty, 103 - Obviousness, 112 - Written Description/Enablement, Double Patenting

CLAIMS:
Amendments to the Claims:This listing of claims replaces all prior versions and listings of claims in the application:Listing of Claims:
(Currently Amended) An antibody or an antigen-binding fragment thereof, comprising three light chain complementarity determining regions and/or three heavy chain complementarity determining regions, wherein
(1) the three light chain complementarity determining regions of the antibody or the antigenbinding fragment thereof comprise an LCDRI set forth in SEQ ID NO: 5, an LCDR2 set forth in SEQ ID NO: 6 and an LCDR3 set forth in SEQ ID NO: 7, and/or the three heavy chain complementarity determining regions of the antibody or the antigen-binding fragment thereof comprise an HCDRI set forth in SEQ ID NO: 8, an HCDR2 set forth in SEQ ID NO: 9 and an HCDR3 set forth in SEQ ID NO: 10,
or
(2) the three light chain complementarity determining regions of the antibody or the antigenbinding fragment thereof comprise an LCDR1 set forth in SEQ ID NO: 11, an LCDR2 set forth in SEQ ID NO: 12 and an LCDR3 set forth in SEQ ID NO: 13, and/or the three heavy chain complementarity determining regions of the antibody or the antigen-binding fragment thereof comprise an HCDRI set forth in SEQ ID NO: 14, an HCDR2 set forth in SEQ ID NO: 15 and an HCDR3 set forth in SEQ ID NO: 16.
(Currently Amended) The antibody or an antigen-binding fragment thereof of claim 1, wherein the antibody or the antigen-binding fragment thereof comprises a light chain variable region set forth in SEQ ID NO: 1, and/or a heavy chain variable region set forth in SEQ ID NO: 2,
or
First Named Inventor : Application No. Filed Page
Deyong SONG 17/601,765 October 6, 2021 4of7
Attorney Docket No.: 48644-0004US1
the antibody or the antigen binding fragmen thereof compises a light chain variable region set forth in SEQ ID NO:3,and/or a heavy chain variable region set forth in SEQ ID NO:4
(Currently Amended) The antibody or the antigen-binding fragment thereof of claim 1, further comprising a heavy chain constant region set forth in SEQ ID NO: 17 or a heavy chain constant region set forth in SEQ ID NO: 18.
(Currently Amended) A chimeric antigen receptor (CAR), comprising the antibody or the antigen-binding fragment thereof of claim 1.
(Currently Amended) The chimeric antigen receptor of claim 4, sequentially comprising the antibody or the antigen-binding fragment thereof, an extracellular hinge region, a transmembrane region and an intracellular signaling region, wherein, preferably, the extracellular hinge region is a CD8 hinge region, the transmembrane region is a CD8 transmembrane region, and the intracellular signaling region is 4-1BB and CD36
6.(Currently Amended) A nucleic acid, wherein the nucleic acid encodes the antibody or the antigen-binding fragment thereof of claim 1, or a chimeric antigen receptor comprising the antigen-binding fragment of any one of claims 1 to3, or the chimeric antigen receptor of claims 4 or 5; and the cell expresses the antibody or the antigen binding fragment thereof of any one of claims 1 to 3, or the chimeric antigen receptor of claims 4 or5.
(Currently Amended) A pharmaceutical composition, comprising the antibody or the antigen-binding fragment thereof of any one of claims 1 to 3, the chimeric antigen receptor according to claims 4 or 5, or the nucleic acid or the cell according to claim 6.
8.(Cancelled)
(Currently Amended) Use of the antibody or the antigen binding fragment thereof
First Named Inventor : Application No. Filed Page
Deyong SONG 17/601,765 October 6, 2021 5 of7
Attorney Docket No.: 48644-0004US1
of any one of claims 1 to3, the chimeric antigen receptor according to claims 4 or 5,or the nucleic acid of claim 6 for treating, preventing, detecting or diagnosing a disease, wherein, preferably, the disease is cancer. A method of treating or preventing cancer, the method comprising administering to a subject in need thereof the antibody or antigen-binding fragment thereof of claim 1.
(Currently Amended) The method of use according to claim 9, wherein the cancer is a Claudin18.2-positive cancer; preferably, the cancer includes gastric cancer, pancreatic cancer, esophageal cancer, lung cancer, ovarian cancer, head and neck cancer, bladder cancer, cervical cancer, sarcoma, cytoma, colon cancer, kidney cancer, colorectal cancer, liver cancer, melanoma, breast cancer, myeloma, neuroglioma, leukemia and lymphoma.
(New) The antibody or antigen-binding fragment thereof of claim 1, wherein the three light chain complementarity determining regions of the antibody or the antigen-binding fragment thereof comprise an LCDRI set forth in SEQ ID NO: 5, an LCDR2 set forth in SEQ ID NO: 6 and an LCDR3 set forth in SEQ ID NO: 7, and/or the three heavy chain complementarity determining regions of the antibody or the antigen-binding fragment thereof comprise an HCDR1 set forth in SEQ ID NO: 8, an HCDR2 set forth in SEQ ID NO: 9 and an HCDR3 set forth in SEQ ID NO: 10.
(New) The antibody or antigen-binding fragment thereof of claim 1, wherein the three light chain complementarity determining regions of the antibody or the antigen-binding fragment thereof comprise an LCDRI set forth in SEQ ID NO: 11, an LCDR2 set forth in SEQ ID NO: 12 and an LCDR3 set forth in SEQ ID NO: 13, and/or the three heavy chain complementarity determining regions of the antibody or the antigen-binding fragment thereof comprise an HCDR1 set forth in SEQ ID NO: 14, an HCDR2 set forth in SEQ ID NO: 15 and an HCDR3 set forth in SEQ ID NO: 16.
13.(New) The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody or the antigen-binding fragment thereof comprises a light chain variable region set forth in SEQ
First Named Inventor : Application No. Filed Page
Deyong SONG 17/601,765 October 6, 2021 6of7
Attorney Docket No.: 48644-0004US1
ID NO: 3, and/or a heavy chain variable region set forth in SEQ ID NO: 4.
(New) The antibody or antigen-binding fragment thereof of claim 1, further comprising a heavy chain constant region set forth in SEQ ID NO: 18
(New) The chimeric antigen receptor of claim 5, wherein the extracellular hinge region is a CD8 hinge region, the transmembrane region is a CD8 transmembrane region, and the intracellular signaling region is 4-1BB and CD36
(New) A cell, wherein the cell expresses the antibody or antigen-binding fragment thereof of claim 1.
(New) A cell, wherein the cell expresses the CAR of claim 4.
(New) A cell, wherein the cell comprises the nucleic acid of claim 6.
(New) A method of treating or preventing cancer, the method comprising administering to a subject in need thereof the cell of claim 17.
(New) A method of detecting Claudin18.2, the method comprising contacting a sample with the antibody or antigen-binding fragment thereof of claim 1.
(New) The method of claim 9, wherein the cancer is selected from the group consisting of gastric cancer, pancreatic cancer, esophageal cancer, lung cancer, ovarian cancer, head and neck cancer, bladder cancer, cervical cancer, sarcoma, cytoma, colon cancer, kidney cancer, colorectal cancer, liver cancer, melanoma, breast cancer, myeloma, neuroglioma, leukemia and lymphoma.

OFFICE ACTION:
DETAILED ACTION
Notice of Pre-AIA  or AIA  Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Status of the Claims
Claims 1-7 and 9-21 are currently pending and are subject to this Office Action. This is the first Office Action on the merits of the claims.
Information Disclosure Statement
The references cited on the information disclosure statement(s) were considered and have been made of record to the extent that each was provided.
Claim Rejections - 35 USC § 112(a)
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
Claims 1-7 and 9-21 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA  35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
Claims 1-2 and 11-12 recite “wherein the antibody… comprises three light chain complementarity determining regions and/or three heavy chain complementarity determining regions,” and claim 13 recites “wherein the antibody… comprises a light chain variable region set forth in SEQ ID NO: 3, and/or a heavy chain variable region set forth in SEQ ID NO: 4”. The specification only teaches antibodies comprising both heavy and light chain variable regions (see, e.g., Table 1), and does not provide any guidance on antibodies only comprising one or the other, and thus, lacks evidence that the full genus of the antibodies encompassed by the present claims (e.g., antibodies that comprise only light chain CDRs) is contemplated. Moreover, it is well understood in the art that conventional antibodies comprise 6 distinct CDR loops, three heavy and three light chain CDRs, which form the antigen binding site and determine antibody diversity and antigen specificity (see, e.g., Chailyan. FEBS J. 2011 Aug;278(16):2858-66; and Chiu. Antibodies 2019, 8(4), 55). Therefore, claims 1-2 and 11-13, and those claims that depend from 1-2 and 11-13, contains subject matter that lacks adequate description in the specification as to reasonably convey to one skilled in the art that the inventor had possession of the claimed invention.
Claims 9-10, 19, and 21 are rejected under 35 U.S.C. 112(a) because the specification, while being enabling for treating cancer, does not reasonably provide enablement for preventing cancer.  The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to use the invention commensurate in scope with these claims.
While the specification does not provide a definition for the terms “treating” or “preventing”, the term “prevention” of disease is widely interpreted in the art as stopping disease from happening entirely. Although Claudin 18.2 is known to be a promising target for the treatment of various cancers and antibodies targeting Claudin 18.2 have been developed to treat patients with Claudin 18.2+ advanced stage gastric cancer (see background of the instant specification, p. 1 par. 2-4 in particular), there is insufficient evidence to show that it can be targeted to prevent cancer from happening entirely. Moreover, due to the nature of antibody therapies, continued administration of the therapy is required to consistently block the target in order to stop the related disease from occurring, such as in applications where antibody therapy is used to prevent infectious viral diseases in high-risk populations [see, e.g., Boonyaratanakornkita. Curr Opin Infect Dis. 2022; 35(4):280-287]. The claims and the specification do not teach how the invention can be used to prevent a disease related to or caused by Claudin 18.2. Therefore, the specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to use the invention commensurate in scope with this claim.
Nature of the invention/Breadth of the claims.  The claims are drawn to a method of treating or preventing cancer.
State of the prior art/Predictability of the art. The prior art teaches Claudin 18.2 targeting as inhibition of further development but not prevention of related cancers.
Working examples. No working example for disease prevention is disclosed in the specification.
Guidance in the specification. The specification provides guidance towards reducing tumor size by blocking NTSR1, but not prevention of a disease, such as cancer.
Amount of experimentation necessary.  Additional research is required in order to determine how effective the invention is at preventing cancer, because the prior art, working examples and guidance in the instant specification do not teach prevention.
For the reasons discussed above, it would require undue experimentation for one skilled in the art to use the claimed methods.
Priority
Receipt is acknowledged of the certified copy of the Foreign Priority Application required by 37 CFR 1.55.
Should applicant desire to obtain the benefit of foreign priority under 35 U.S.C. 119(a)-(d) prior to declaration of an interference, a certified English translation of the foreign application must be submitted in reply to this action.  37 CFR 41.154(b) and 41.202(e).
Failure to provide a certified translation may result in no benefit being accorded for the non-English application.
Claim Rejections - 35 USC § 102
In the event the determination of the status of the application as subject to AIA  35 U.S.C. 102 and 103 (or as subject to pre-AIA  35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA  to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claims 1, 6-7, 9, 11, 16, 18, and 21 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Lantto (WO2018091661; cited on the IDS filed on 7/8/2022).
Regarding claims 1 and 11, Lantto discloses an anti-PD-1 antibody or antigen-binding portion thereof for enhancing immunity or treating cancer (for example, non-small cell lung cancer), the amino acid sequence as described in SEQ ID NO: 10 comprising the same LCDRs as those in SEQ ID NOs: 5-7 of the present application (see abstract, claims 1 and 56-57, and SEQ ID NO: 10).
Regarding claim 6, Lantto discloses an isolated nucleic acid molecule comprising a nucleotide sequence that encodes the anti-PD-1 antibody (see claim 51).
Regarding claim 7, Lantto discloses a pharmaceutical composition comprising the anti-PD-1 antibody (see claim 49).
Regarding claim 9, Lantto discloses a method of treating cancer in a patient comprising administering the anti-PD-1 antibody (see claim 57).
Regarding claim 18, Lantto discloses a host cell comprising the nucleotide sequence that encodes the anti-PD-1 antibody (see claim 53).
Regarding claim 21, Lantto discloses the method of treating cancer, wherein the cancer is one of those listed in instant claim 21 (see claim 58-59).
Thus, Lantto anticipates the subject matter of instant claims 1, 6-7, 9, 11, 16, 18, and 21.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA  35 U.S.C. 102 and 103 (or as subject to pre-AIA  35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA  to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary.  Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 4-5, 15, 17, and 19 are rejected under 35 U.S.C. 103 as being unpatentable over Lantto (supra) in view of Almasbak (Treatment, Journal of Immunology Research, 2016, 5474602, 10 pages, 2016.) and Li (US20190117691).
The disclosures of Lantto are discussed above and are incorporated herein.
Lantto fails to disclose the chimeric antigen receptor (CAR) of the present invention.
Regarding claim 4:
Almasbak teaches: “despite recent therapeutic advances, such as the introduction of monoclonal antibodies and small-molecular inhibitors, treatment responses vary considerably among patients and a high relapse rate with poor prognosis continues to be a major challenge… Overwhelming evidence supports the critical role of the immune system, and lymphocytes in particular, in controlling and eradicating cancer” (see p. 1 col. 1) and “years of successive and significant innovations have finally culminated in clinical studies demonstrating the tremendous potential of second generation CAR expressing T cells” (see p. 2 col. 1).
Li discloses a PD-1 CAR-T cell, a preparation method thereof, and an application thereof (see abstract and claims).
In light of these teachings, it would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have used the antibody of Lantto to develop a CAR comprising the same CDR sequences, as CAR technology is well established in the art as a superior treatment for cancer. One would have been motivated specifically use the antibody of Lantto to develop a CAR, as PD-1 is known to be associated with cancers. Therefore, if an artisan wanted to develop a treatment for cancer, particularly cancers associated with PD-1, one would have been motivated to develop a CAR comprising the antibody of Lantto.
Regarding claims 5, 15, and 17, Li discloses a PD-1 CAR-T cell expressing a PD-1-CD8-4-1 BB-CD3ζ fusion protein, wherein the PD-1 portion of the fusion protein is expressed on a surface of the CAR-T cell and the 4-1 BB-CD3ζ portion of the fusion protein is expressed inside the CAR-T cell (see claim 1; reads on instant claims 5, 15, and 17).
Regarding claims 19, Li discloses a method of treating tumor by administering to a patient a PD-1 CAR-T cell (see claim 22; reads on instant claim 19).
Claim 3 is rejected under 35 U.S.C. 103 as being unpatentable over Lantto (supra) in view of Parren (WO2013004841).
The disclosures of Lantto are discussed above and are incorporated herein.
Regarding claim 3, Lantto fails to disclose the heavy chain constant region sequence of instant SEQ ID NO: 17. However, Parren discloses heavy chain constant region sequence of SEQ ID NO: 331, which shares 100% identity with instant SEQ ID NO: 17.
In light of these teachings, it would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the antibody of Lantto to incorporate the heavy chain constant region sequence of Parren, as this sequence is known in the prior art, and doing so amounts to no more than simple substitution of one known element for another to obtain predictable results. See MPEP § 2143.
Double Patenting
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the conflicting claims are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on nonstatutory double patenting provided the reference application or patent either is shown to be commonly owned with the examined application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA  as explained in MPEP § 2159. See MPEP § 2146 et seq. for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The filing of a terminal disclaimer by itself is not a complete reply to a nonstatutory double patenting (NSDP) rejection. A complete reply requires that the terminal disclaimer be accompanied by a reply requesting reconsideration of the prior Office action. Even where the NSDP rejection is provisional the reply must be complete. See MPEP § 804, subsection I.B.1. For a reply to a non-final Office action, see 37 CFR 1.111(a). For a reply to final Office action, see 37 CFR 1.113(c). A request for reconsideration while not provided for in 37 CFR 1.113(c) may be filed after final for consideration. See MPEP §§ 706.07(e) and 714.13.
The USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/patent/patents-forms. The actual filing date of the application in which the form is filed determines what form (e.g., PTO/SB/25, PTO/SB/26, PTO/AIA /25, or PTO/AIA /26) should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to www.uspto.gov/patents/apply/applying-online/eterminal-disclaimer.
Copending Application No. 18/019,179
Claims 1, 4-6, 12-13, 15, and 17-19 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-20 of copending Application No. 18/019,179 (reference application). Although the claims at issue are not identical, they are not patentably distinct from each other because the claims of Application ‘179 anticipate the subject matter of the present claims.
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
Regarding claims 1, 4-5, and 12-13, Application ‘179 claims a CAR comprising an scFv, an extracellular hinge, a transmembrane domain, and an intracellular domain, wherein the scFv is an anti-Claudin 18.2 comprising SEQ ID NO: 30 (see claims 1-2 in particular). SEQ ID NO: 30 comprises instant SEQ ID NOs: 3-4 and 11-16.
Regarding claim 6, Application ‘179 claims the nucleic acid encoding the CAR (see claim 9 in particular).
Regarding claim 15, Application ‘179 claims the CAR comprising a CD8 hinge, CD8 transmembrane domain, and intracellular domain of 41BB and CD3 zeta (see claim 5 in particular).
Regarding claim 17-18, Application ‘179 claims the cell expressing the CAR and nucleic acid (see claim 14).
Regarding claim 19, Application ‘179 claims the method of treating cancer comprising administering the CAR cell (see claim 12)
Thus, Application ‘179 anticipates the subject matter of the present claims.
Conclusion
Claims 1-7 and 9-21 are rejected.


NEXT OFFICE ACTION: DETAILED ACTION
Applicant’s reply filed on 12/4/2024 is acknowledged and entered.
Applicant’s supplemental response filed on 2/7/2025 is entered.
Claims 1-7 and 9-21 are pending. Claim 8 is canceled.
Claims 1-7 and 9-21 are allowed.
EXAMINER’S AMENDMENT
An examiner’s amendment to the record appears below. Should the changes and/or additions be unacceptable to applicant, an amendment may be filed as provided by 37 CFR 1.312. To ensure consideration of such an amendment, it MUST be submitted no later than the payment of the issue fee.
Authorization for this examiner’s amendment was given in an interview with Applicant’s representative Jia Zhu on Feb 4, 2025.
The application has been amended as follows:
Amendments to the Claims
In claim 1, 	line 1, after the term “thereof”, the phrase --which binds to Claudin18.2-- has 			been added.
In claim 16, 	line 1, the term “A cell” has been changed to --An isolated cell--.
In claim 17, 	line 1, the term “A cell” has been changed to --An isolated cell--.
In claim 18, 	line 1, the term “A cell” has been changed to --An isolated cell--.
In claim 19, 	line 2, after the term “claim 17”, the phase --, wherein the cell is a T cell or 			NK cell-- has been added.
REASONS FOR ALLOWANCE
6.	The 35 USC 112(a) and art rejections in the last office action mailed on 9/10/2024 are withdrawn in view of applicant’s amendments.
The nonstatutory double patenting rejection is withdrawn as the instant application has an earlier patent term filing date.
7.

CITED REFERENCE SUMMARIES: Ref US20190117691A1: [content unavailable]
Ref WO2013004841A1: Claims:
A method for producing an antibody in a host cell, wherein said antibody comprises at least a heavy chain, said method comprising the following steps: a) providing a nucleotide construct encoding said heavy chain, wherein said construct does not encode a lysine residue at the C-terminus of said heavy chain, b) expressing said nucleotide construct in a host cell, with the proviso that said host cell is not a CHO cell or a moss cell, and c) recovering said antibody from a cell culture of said host cell.
The method according to claim 1, wherein said host cell further expresses a light-chain-encoding construct.
The method according to claim 1 or 2, wherein said host cell is capable of Asn-linked glycosylation of proteins.
The method according to any one of the preceding claims 1 to 3, wherein said host cell is a non-human cell which is genetically engineered to produce glycoproteins having human-like or human glycosylation.
The method according to any one of the preceding claims, wherein said host cell is a host cell which is not capable of efficiently removing C-terminal lysine residues from antibody heavy chains.
The method according to claim 5, wherein said host cell is a host cell which, when expressing a C-terminal-lysine-containing construct, would produce an antibody preparation wherein more than 10% of the antibody molecules is in the K2 isoform, e.g. wherein more the 30% is in the K2 isoform.
The method according to any one of the preceding claims, wherein said host cell is selected from the group consisting of:  a) a yeast cell, e.g. a Pichia pastoris or Saccharomyces cerevisiae, Hansenula polymorpha and Ogataea minuta, b) a filamentous fungal cell, such as Aspergillus awamori, Aspergillus niger, Aspergillus oryzea, Trichoderma reesi, c) a plant cell, such as an Arabidopsis cell, Lemna minor, Nicotiana benthamiana (tobacco), oilseed crop plant (Brassica napus), soybean, rice, maize (Zea mays) or carrot cells, d) an NSO cell, an Sp2/0 cell, and a PER.C6 cell.
The method according to any one of the preceding claims, wherein the nucleotide construct provided in step a) is derived from, or designed based on, an original heavy-chain sequence having a codon for a C-terminal lysine residue, e.g. wherein said nucleotide construct comprises a deletion or a substitution of the codon for the C-terminal lysine residue compared to said original heavy-chain sequence.
The method according to any one of the preceding claims, wherein said heavy-chain-encoding construct does not encode a lysine or an arginine at the C- terminus of said heavy chain, preferably wherein said heavy-chain-encoding construct does not encode a charged amino acid residue at the C-terminus of said heavy chain.
The method according to any one of the preceding claims, wherein said nucleotide construct, other than the C-terminal modification, encodes a heavy chain of the IgGl or IgG2 isotype.
The method according to any one of the preceding claims, wherein the C- terminal codon of said nucleotide construct encodes a Pro or a Gly residue.
An isolated antibody obtained or obtainable by the method of any one of the preceding claims.
A host cell which is not capable of efficiently removing C-terminal lysine residues from antibody heavy chains, said host cell comprising a nucleotide construct encoding a heavy chain lacking a C-terminal lysine.
The host cell according to claim 13, wherein said host cell is of one of the types specified in claim 7.
A method for increasing the ability of an antibody to mediate complement- dependent cytotoxicity, said method comprising the step of removing a C-terminal lysine residue from the heavy-chains of an antibody.
The method according to claim 15, wherein the lysine residue is removed by enzymatic cleavage, e.g. using a carboxypeptidase.
The method according to claim 15 or 16, wherein said antibody is purified prior to removal of the C-terminal lysine residue.
The method according to any one of claim 15 to 17, wherein said antibody is of the IgGl or IgG2 isotype.
A method for testing a plurality of antibody-producing cell cultures, e.g. hybridoma cell cultures, for a desired property, said method comprising treating samples of cultures of said hybridomas with the method according to any one of claims 15 to 18 and testing the treated samples for the desired property.
The method according to claim 19, wherein the desired property is cell lysis or cell killing.
An isolated antibody variant comprising a heavy chain having at least one charged amino acid residue in the C-terminal region, wherein said charged amino acid residue is not susceptible to proteolytic removal.
The antibody variant according to claim 21 further comprising a light chain.
The antibody variant according to any one of the preceding claims 21 to 22, wherein the antibody has decreased ability to mediate complement-dependent cytotoxicity compared to an antibody lacking said charged amino acid residue.
The antibody variant according to any one of the preceding claims 21 to 23, wherein said charged amino acid residue is not susceptible to removal by proteases of mammalian origin, preferably not susceptible to removal by proteases of CHO, HEK-293, PER.C6, NSO or Sp2/0 origin, more preferably not susceptible to removal  by proteases active in the secretory pathway of CHO, HEK-293, PER.C6, NSO or Sp2/0 cells.
The antibody variant according to any one of the preceding claims 21 to 24, wherein said charged amino acid residue is not susceptible to removal by human proteases, preferably not susceptible to removal by human proteases that can act on proteins in circulation.
The antibody variant according to any one of the preceding claims 21 to 25, wherein said charged amino acid is amongst the six most C-terminal amino acid residues, e.g. amongst the five most C-terminal amino acid residues of said heavy chain, such as amongst the four most C-terminal amino acid residues of said heavy chain, e.g. amongst the three most C-terminal amino acid residues of said heavy chain, such as amongst the two most C-terminal amino acid residues of said heavy chain, e.g. wherein said charged amino acid is the most C-terminal amino acid residue of said heavy chain.
The antibody variant according to any one of the preceding claims 21 to 26, wherein said heavy chain comprises the CH3 sequence set forth in SEQ ID NO: l, 2, 3 or 5 and said charge amino acid is situated between positions 325-330, 321-326, 372-377 and 325-330, respectively.
The antibody variant according to any one of the preceding claims 21 to 27, wherein said heavy chain comprises the entire constant region sequence set forth in
SEQ ID NO: l, 2, 3 or 5 and said charge amino acid is situated between positions 325-330, 321-326, 372-377 and 325-330, respectively.
The antibody variant according to any one of the preceding claims 21 to 28, wherein said charged amino acid is a positively charged amino acid residue, preferably a lysine residue.
The antibody variant according to any one of the preceding claims 21 to 28, wherein said charged amino acid is a negatively charged amino acid residue, preferably a glutamic acid residue.
The antibody variant according to any one of the preceding claims 21 to 30, wherein said charged amino acid residue is not susceptible to proteolytic removal  due to amino acid modifications at positions situated C-terminally of said charged amino acid residue.
The antibody variant according to any one of the preceding claims 21 to 31, wherein said charged amino acid residue is not susceptible to proteolytic removal due to the presence of a proline residue situated C-terminally of said charged amino acid residue, preferably wherein said proline residue is placed immediately C- terminal of said charged amino acid residue, more preferably wherein said charged amino acid residue and said proline residue are the two most C-terminal amino acid residues of said heavy chain.
The antibody variant according to any one of the preceding claims 21 to 32, wherein said heavy chain comprises two or more charged amino acid residues in the C-terminal region, such as two, three, four or five charged amino acid residues, preferably having either all a positive charge or all a negative charge.
The antibody variant according to any one of the preceding claims 21 to 33, wherein the specified amino acid modifications are the result of amino acid substitutions.
The antibody variant according to any one of the preceding claims 21 to 34, wherein the specified amino acid modifications are the result of C-terminal amino acid additions.
The antibody variant according to any one of the preceding claims 21 to 35, wherein said antibody is a human antibody, preferably a human IgGl or a human IgG3 antibody.
The antibody variant according to any one of the preceding claims 21 to 36, wherein said antibody heavy chain is an IgGl, IgG2, IgG3 or IgG4 heavy chain to which a proline residue has been added at the C-terminus.
The antibody variant according to any one of the preceding claims 21 to 37, wherein said antibody is not conjugated at the C-terminus.
An antibody according to any one of claims 21 to 38 for use as a medicament.
A nucleotide construct encoding a heavy chain having a charged amino acid residue in the C-terminal region, wherein said charged amino acid residue is not susceptible to proteolytic removal.
The nucleotide construct according to claim 40, having the further features of any one of claims 22 to 38.
A host cell capable of producing an antibody variant according to any one of claims 21 to 38.
A host cell which has been genetically modified to eliminate the activity of carboxypeptidases capable of removing C-terminal lysines from heavy chains.
The host cell of claim 43, wherein said cell is a CHO, HEK-293, PER.C6, NS0 or Sp2/0 cell.
A method for producing an antibody variant according to any one of claims 21 to 38 comprising culturing the host cell according to any one of claims 42 to 44 and recovering said antibody from the cell culture.
The method according to claim 45, wherein said host cell is a CHO or HEK cell comprising a nucleotide-construct encoding a heavy chain having a negatively- charged amino acid residue at the C-terminus or a proline residue preceded by a positively-charged amino acid residue at the C-terminus.
An antibody mixture comprising a subpopulation of antibody variant molecules comprising heavy chains having at least one positively-charged amino acid residue in the C-terminal region and a subpopulation of antibody variants molecules comprising heavy chains having at least one negatively-charged amino acid residue C-terminal region, wherein said positively-charged and negatively-charged amino acid residues are not susceptible to proteolytic removal.
The antibody mixture according to claim 47, wherein each of said subpopulations constitutes at least 10%, such as at least 20% of said mixture, e.g. at least 30% of said mixture.
The antibody mixture according to claim 47 or 48, wherein said antibody molecules have the properties specified in one or more of claims 22 to 38.
The antibody mixture according to any one of the preceding claims 47 to 49, wherein the antibody molecules within each of the two subpopulations comprise two identical heavy chains.
The antibody mixture according to any one of the preceding claims 47 to 50 for use as a medicament, preferably for use in the treatment of cancer.
An isolated antibody comprising a heavy chain having a variant C-terminal region, comprising one or more amino acid modifications that favour intermolecular C-terminal interactions between antibody molecules.
The antibody according to claim 52, wherein the antibody has increased ability to mediate complement-dependent cytotoxicity compared to an antibody lacking said amino acid modifications.
The antibody according to claim 52, wherein said antibody comprises two non-identical heavy chains.
The antibody according to claim 52, wherein one heavy chain comprises a negatively-charged amino acid residue C-terminal region and the other heavy chain comprises a positively-charged charged amino acid residue in the C-terminal region, wherein said charged amino acid residues are not susceptible to proteolytic removal.
The antibody according to any one of the preceding claims 52 to 55, wherein the antibody is a monospecific antibody.
The antibody according to any one of the preceding claims 52 to 55, wherein the antibody is a bispecific antibody.
The antibody according to any one of the preceding claims 52 to 57 for use as a medicament.
A method for producing an antibody according to any one of claims 52 to 57 comprising culturing a host cell comprising nucleotide constructs encoding said heavy chains and recovering said antibody from the cell culture of said host cell.
The method according to claim 59, wherein said host cell is a CHO or HEK cell.
Ref US20190117691: Claims:
1-9. (canceled)
10. A PD-1 CAR-T cell expressing a PD-1-CD8-4-1 BB-CD3ζ fusion protein, wherein the PD-1 portion of the fusion protein is expressed on a surface of the CAR-T cell and the 4-1 BB-CD3ζ portion of the fusion protein is expressed inside the CAR-T cell.
11. The PD-1 CAR-T cell of claim 10, wherein the amino acid sequence of PD-1 in the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:5; and the amino acid sequence of CD8 in the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:1.
12. The PD-1 CAR-T cell of claim 10, wherein the amino acid sequence of 4-1BB in the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:2; wherein the 4-1BB in the PD-1-CD8-4-1 BB-CD3ζ fusion protein can be replaced by CD28 that has the amino acid sequence of SEQ ID NO:3.
13. The PD-1 CAR-T cell of claim 10, wherein the amino acid sequence of CD3ζ in the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:4.
14. The PD-1 CAR-T cell of claim 10, wherein the amino acid sequence of the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:6.
15. The PD-1 CAR-T cell of claim 10, wherein the T cell is derived from human periphery blood T lymphocytes.
16. A method of making a PD-1 CAR-T cell, comprising:
(1) cloning a gene encoding a PD-1-CD8-4-1 BB-CD3ζ fusion protein into a lentiviral expression vector; (2) packaging and preparing a lentivirus by expressing lentiviral envelop plasmid and the lentiviral expression vector of step (1) in a 293 T cell; (3) isolating and expanding human peripheral blood T lymphocytes, and infecting the T lymphocytes with the lentivirus of step (2) to obtain the PD-1 CAR-T cells expressing the PD-1-CD8-4-1 BB-CD3ζ fusion protein; wherein the PD-1 portion of the fusion protein is expressed on a surface of the CAR-T cell and the 4-1 BB-CD3ζ portion of the fusion protein is expressed inside the CAR-T cell.
17. The method of claim 16, wherein the amino acid sequence of PD-1 in the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:5; and the amino acid sequence of CD8 in the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:1.
18. The method of claim 16, wherein the amino acid sequence of 4-1 BB in the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:2; wherein the 4-1 BB in the PD-1-CD8-4-1BB-CD3ζ fusion protein can be replaced by CD28 that has the amino acid sequence of SEQ ID NO:3.
19. The method of claim 16, wherein the amino acid sequence of CD3ζ in the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:4.
20. The method of claim 16, wherein the amino acid sequence of the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:6.
21. The method of claim 16, wherein the T cell is derived from human periphery blood T lymphocytes.
22. A method of treating tumor by administering to a patient a PD-1 CAR-T cell expressing a PD-1-CD8-4-1 BB-CD3ζ fusion protein; wherein the PD-1 portion of the fusion protein is expressed on a surface of the CAR-T cell and the 4-1 BB-CD3ζ portion of the fusion protein is expressed inside the CAR-T cell.
23. The method of claim 22, wherein the amino acid sequence of PD-1 in the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:5; and the amino acid sequence of CD8 in the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:1.
24. The method of claim 22, wherein the amino acid sequence of 4-1 BB in the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:2; wherein the 4-1 BB in the PD-1-CD8-4-1BB-CD3ζ fusion protein can be replaced by CD28 that has the amino acid sequence of SEQ ID NO:3.
25. The method of claim 22, wherein the amino acid sequence of CD3ζ in the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:4.
26. The method of claim 22, wherein the amino acid sequence of the PD-1-CD8-4-1 BB-CD3ζ fusion protein is SEQ ID NO:6.
27. The method of claim 22, wherein the tumor is characterized with high expression level of PDL-1.
Ref WO2018091661: Claims:
Claims 1 . An anti-PD-1 antibody or an antigen-binding portion thereof, wherein the antibody competes for binding to human PD-1 with, or binds to the same epitope of human PD-1 as, antibody 18040, 18049, 18098, 181 13, 18201 , 18247, 18250, 18325, 18366, 18400, 18413 or 18483. 2. The anti-PD-1 antibody or antigen-binding portion of claim 1 , wherein the anti- PD-1 antibody is selected from the group consisting of: a) an antibody whose H-CDR1 -3 comprise the amino acid sequences of SEQ ID NOs: 29-31 , respectively; b) an antibody whose heavy chain variable domain (VH) is at least 90% identical in sequence to the amino acid sequence of SEQ ID NO: 1 ; c) an antibody whose VH comprises the amino acid sequence of SEQ ID NO: 1 ; d) an antibody whose heavy chain (HC) comprises the amino acid sequences of SEQ ID NOs: 1 and 26; e) an antibody whose L-CDR1 -3 comprise the amino acid sequences of SEQ ID NOs: 32-34, respectively; f) an antibody whose light chain variable domain (VL) is at least 90% identical in sequence to the amino acid sequence of SEQ ID NO: 2; g) an antibody whose VL comprises the amino acid sequence of SEQ ID NO: 2; h) an antibody whose light chain (LC) comprises the amino acid sequences of SEQ ID NOs: 2 and 28; i) an antibody whose H-CDR1 -3 and L-CDR1 -3 comprise the amino acid sequences of SEQ ID NOs: 29-34, respectively; j) an antibody whose VH is at least 90% identical in sequence to the amino acid sequence of SEQ ID NO: 1 and whose VL is at least 90% identical in sequence to the amino acid sequence of SEQ ID NO: 2; k) an antibody whose VH comprises the amino acid sequence of SEQ ID NO: 1 and whose VL comprises the amino acid sequence of SEQ ID NO: 2; and I) an antibody whose HC comprises the amino acid sequences of SEQ ID NOs: 1 and 26 and whose LC comprises the amino acid sequences of SEQ ID NOs: 2 and 28. 3. The anti-PD-1 antibody or antigen-binding portion of claim 1 , wherein the anti- PD-1 antibody is selected from the group consisting of: a) an antibody whose H-CDR1 -3 comprise the amino acid sequences of SEQ ID NOs: 35-37, respectively; b) an antibody whose heavy chain variable domain (VH) is at least 90% identical in sequence to the amino acid sequence of SEQ ID NO: 3; c) an antibody whose VH comprises the amino acid sequence of SEQ ID NO: 3; d) an antibody whose heavy chain (HC) comprises the amino acid sequences of SEQ ID NOs: 3 and 26; e) an antibody whose L-CDR1 -3 comprise the amino acid sequences of SEQ ID NOs: 38-40, respectively; f) an antibody whose light chain variable domain (VL) is at least 90% identical in sequence to the amino acid sequence of SEQ ID NO: 4; g) an antibody whose VL comprises the amino acid sequence of SEQ ID NO: 4; h) an antibody whose light chain (LC) comprises the amino acid sequences of SEQ ID NOs: 4 and 28; i) an antibody whose H-CDR1 -3 and L-CDR1 -3 comprise the amino acid sequences of SEQ ID NOs: 35-40, respectively; j) an antibody whose VH is at least 90% identical in sequence to the amino acid sequence of SEQ ID NO: 3 and whose VL is at least 90% identical in sequence to the amino acid sequence of SEQ ID NO: 4; k) an antibody whose VH comprises the amino acid sequence of SEQ
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