CAR-iNKT CELL TECHNOLOGY EFFECTIVE IN KILLING CHOLANGIOCARCINOMA

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 . DETAILED ACTION Applicant’s election without traverse of Group I, claims 1-14 and 26, in the reply filed on 07/28/2026 is acknowledged. Claims 1-14, 16-20, and 26 are pending. Claims 16-20 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 07/28/2026. Claims 1-14 and 26 are under examination on the merits. Priority Applicant’s claim for the benefit of a prior-filed application under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) is acknowledged. Claims 1-14 and 26 have an effective filing date of 01/20/2023. Information Disclosure Statement The information disclosure statements (IDS) submitted on 02/12/2025 and 06/17/2025 are in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statements are being considered by the examiner. Claim Rejections 35 U.S.C. 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 1-5, 10, 12, and 13 are rejected under 35 U.S.C. 103 as being unpatentable over Genkin et al. (WO 2020/212756, publication date: 10/22/2020) in view of Qian et al. (WO 2019/020088, publication date: 01/31/2019). At p. 1 and 2, Genkin et al. (WO 2020/212756, publication date: 10/22/2020) teach that “[t]he present invention provides improved cancer immunotherapies utilizing polymorphonuclear leukocytes (PMNs). In one aspect, provided herein is a method of treating a cancer in a subject in need thereof comprising administering to the subject a therapeutically effective amount of polymorphonuclear leukocytes, wherein the polymorphonuclear leukocytes are genetically modified to express a recombinant chimeric antigen receptor (CAR) or T cell receptor (TCR). In some embodiments, the polymorphonuclear leukocytes express a chimeric antigen receptor (CAR) targeting 4-1BB, 5T4, adenocarcinoma antigen, alpha-fetoprotein, BAFF, B-lymphoma cell, C242 antigen, CA-125, carbonic anhydrase 9 (CA-IX), C-MET, CCR4, CD 152, CD 19, CD20, CD200, CD22, CD221, CD23 (IgE receptor), CD28, CD30 (TNFRSF8), CD33, CD4, CD40, CD44 v6, CD51, CD52, CD56, CD74, CD80, CEA, CNT0888, CTLA-4, DR5, EGFR, EpCAM, CD3, FAP, fibronectin extra domain-B, folate receptor 1, GD2, GD3 ganglioside, glycoprotein 75, GPNMB, HER2/neu, HGF, human scatter factor receptor kinase, mesothelin (emphasis added)…” At p. 3, Genkin et al. teach the mesothelin-targeting CAR of SEQ ID NO: 6, which shares 100% sequence homology with the MSLN single-chain antibody of SEQ ID NO: 8. At p. 7, Genkin e al. teach that CARs of the invention may comprise a CD28 transmembrane region and a CD28 co-stimulatory domain, and Genkin et al. teach that CARs of the invention also may comprise a lymphocyte activation domain, i.e., signal region or intracellular signaling domain, from CD3ζ. Therefore Genkin et al. teach a CAR, comprising a MSLN single-chain antibody, a transmembrane region (CD28), an intracellular co-stimulatory domain (CD28), and a signal region (CD3ζ), linked in sequence. Genkin et al. do not teach or suggest a CAR, comprising a MSLN single-chain antibody, a spacer domain or hinge region, a transmembrane region (CD28), an intracellular co-stimulatory domain (CD28), and a signal region (CD3ζ), linked in sequence. These deficiencies are remedied by Qian et al. Qian et al. teach CARs that target mesothelin, see throughout, and at [0032], Qian et al. teach a CAR that comprises a CD8 signal peptide, an anti-MSLN single chain antibody, a CD8 hinge region, a CD8 transmembrane region, a CD28 intracellular domain, and an intracellular activation domain from CD3ζ. At [0061], Qian et al. specifically teach a hinge region from CD8α. At [0016], Qian et al. teach the CD8 signal peptide of SEQ ID NO: 1, which comprises the instant SEQ ID NO: 2 (CD8α signal peptide). At [0061], Qian et al. teach the CD8α hinge of SEQ ID NO: 5, which comprises the instant SEQ ID NO: 14 (CD8α hinge). One of ordinary skill in the art would have been motivated with a reasonable expectation of success at the effective filing date of the invention to combine the teachings of Genkin et al. and Qian et al. to develop a CAR comprising a MSLN single-chain antibody (the instant SEQ ID NO: 8), a spacer domain or hinge region (CD8), a transmembrane region (CD28), an intracellular co-stimulatory domain (CD28), and a signal region (CD3ζ), linked in sequence. One of ordinary skill in the art would have been motivated to do so, because Genkin et al. teach a CAR, comprising a MSLN single-chain antibody, a transmembrane region (CD28), an intracellular co-stimulatory domain (CD28), and a signal region (CD3ζ), linked in sequence. Furthermore Qian et al. teach that mesothelin-targeting CARs may comprise a CD8α signal peptide and a CD8α hinge region. Therefore one of ordinary skill in the art would have been motivated to modify the invention of Genkin et al. to comprise the CD8α signal peptide and a CD8α hinge region of Qian et al., because there would have been a reasonable expectation that the resultant anti-MSLN CAR is effective in eliminating MSLN-expressing cancer cells. The invention of Genkin et al. and Qian et al. meets the limitations of claims 1-3 and 5. Claim 4 is included in this rejection, because at [0039], Qian et al. teach expression vectors that comprise nucleic acids which can be used to express CARs. As such one of ordinary skill in the art would have been motivated to prepare nucleic acids that encode anti-MSLN CARs comprising the MSLN single-chain antibody of the instant SEQ ID NO: 8, thus arriving at the nucleotide sequences of the instant claim 4. With respect to claims 10, 12 and 13, at Examples 2 and 3, Genkin et al. teach that transduction of leukocytes with vectors that code for a CAR, and at [0004], Qian et al. teach that “[a]s one of the important branches of tumor immunotherapy, chimeric antigen receptor T cell therapy has achieved very good efficacy in malignant hematological tumors, and has a complete remission rate of more than 90% for relapsed refractory B-cell leukemia. Chimeric antigen receptors are an artificial synthetic receptor, which typically comprises an extracellular antigen binding domain, a transmembrane hinge region, and an intracellular signaling region. Gene recombination is performed in vitro by single-chain fragment variable (scFv) and intracellular signal domain ‘immune receptor tyrosine-based activation motif (ITAM)’ identifying tumor associated antigens (TAA). The virus or other vector system is then introduced into T cells, so that the genetically modified T cells are referred to as CAR-T cells. CAR-T cells are subjected to large-scale amplification in vitro and returned to the patient, and the CAR-T cells can exhibit a strong anti-cancer effect in a non-MHC-limiting mode (emphasis added).” Based upon these teachings, one of ordinary skill in the art would have been motivated to transduce T cells with nucleic acids encoding anti-MSLN CARs, because the resultant transduced T cells could be used in the treatment of MSLN-expressing cancers. Therefore the invention as a whole was prima facie obvious to one of ordinary skill in the art at the effective filing date of the invention, as evidenced by the references. Claims 11 and 14 are rejected under 35 U.S.C. 103 as being unpatentable over Genkin et al. (WO 2020/212756, publication date: 10/22/2020) in view of Qian et al. (WO 2019/020088, publication date: 01/31/2019), as applied to claims 1-5, 9, 10, 12, and 13, and further in view of CN112225822 (Publication date: 01/15/2021). As indicated above one of ordinary skill in the art would have been motivated with a reasonable expectation of success at the effective filing date of the invention to combine the teachings of Genkin et al. and Qian et al. to develop a CAR comprising a MSLN single-chain antibody (the instant SEQ ID NO: 8), a spacer domain or hinge region (CD8), a transmembrane region (CD28), an intracellular co-stimulatory domain (CD28), and a signal region (CD3ζ), linked in sequence. With respect to claims 10, 12 and 13, at Examples 2 and 3, Genkin et al. teach that transduction of leukocytes with vectors that code for a CAR, and at [0004], Qian et al. teach that “[a]s one of the important branches of tumor immunotherapy, chimeric antigen receptor T cell therapy has achieved very good efficacy in malignant hematological tumors, and has a complete remission rate of more than 90% for relapsed refractory B-cell leukemia. Chimeric antigen receptors are an artificial synthetic receptor, which typically comprises an extracellular antigen binding domain, a transmembrane hinge region, and an intracellular signaling region. Gene recombination is performed in vitro by single-chain fragment variable (scFv) and intracellular signal domain ‘immune receptor tyrosine-based activation motif (ITAM)’ identifying tumor associated antigens (TAA). The virus or other vector system is then introduced into T cells, so that the genetically modified T cells are referred to as CAR-T cells. CAR-T cells are subjected to large-scale amplification in vitro and returned to the patient, and the CAR-T cells can exhibit a strong anti-cancer effect in a non-MHC-limiting mode (emphasis added).” Based upon these teachings, one of ordinary skill in the art would have been motivated to transduce T cells with nucleic acids encoding anti-MSLN CARs, because the resultant transduced T cells could be used in the treatment of MSLN-expressing cancers. With respect to claim 11, although the references cited teach or suggest a vector carrying a CAR comprising a MSLN single-chain antibody (the instant SEQ ID NO: 8), a spacer domain or hinge region (CD8), a transmembrane region (CD28), an intracellular co-stimulatory domain (CD28), and a signal region (CD3ζ), linked in sequence, these references do not teach or suggest the pLV300 vector comprising the nucleotide sequence of SEQ ID NO: 24; however this deficiency is remedied by CN112225822. At p. 9, CN112225822 teaches that “[t]he invention comprises a nucleic acid expression vector containing one or more elements and used for constructing the chimeric antigen receptor protein expressed on the surface of T cells, NK cells or NKT cells. Various commercially available vectors can be selected as needed, or the vectors can be constructed according to the conventional techniques in the field of molecular biology. In a specific embodiment, the vector used in the present invention is a lentiviral plasmid vector pLV300. This plasmid belongs to a fourth generation self-inactivating lentiviral vector system that has a total of four plasmids, namely a packaging plasmid encoding the protein Gag/Pol, a packaging plasmid encoding the Rev protein, an envelope plasmid encoding the VSV-G protein, and an empty vector pLV300, which can be used for the recombinant introduction of a nucleic acid sequence of interest, i.e. a nucleic acid sequence encoding a chimeric antigen receptor protein.” Based upon these teachings, one of ordinary skill in the art would have been motivated with a reasonable expectation of success at the effective filing date of the invention to prepare the claimed CAR using the pLV300 vector, because CN112225822 teaches that said vector may be used in the preparation of CARs. Furthermore SEQ ID NO: 40 of CN112225822 appears to be the nucleic acid sequence of pLV300, and said sequence shares 100% homology with the instant SEQ ID NO: 24. With respect to claims 14, as indicated above based upon the teachings of the prior art, one of ordinary skill in the art would have been motivated to transduce T cells with nucleic acids encoding anti-MSLN CARs, because the resultant transduced T cells could be used in the treatment of MSLN-expressing cancers. Neither Genkin et al. nor Qian et al. teach or suggest the transduction of iNKT cells with nucleic acids encoding an anti-MSLN CAR. This deficiency is also remedied by CN112225822. At p. 6, CN112225822 teaches that “the applicant constructs anti-GPC3-CAR-iNKT cells which can specifically kill liver cancer cells carrying GPC3 antigen by using the homing property and non-specific killing function of the iNKT cells and carrying chimeric antigen receptors capable of binding GPC3 antigen through gene modification.” Based upon these teachings, one of ordinary skill in the art would have been motivated with a reasonable expectation of success at the effective filing date of the invention to prepare anti-MSLN-CAR-iNKT cells, because there would have been a reasonable expectation that said cells would provide a therapeutic benefit to patients having MSLN-expressing cancers by killing MSLN-expressing cancer cells. Therefore the invention as a whole was prima facie obvious to one of ordinary skill in the art at the effective filing date of the invention, as evidenced by the references. Claims 6 and 7 are rejected under 35 U.S.C. 103 as being unpatentable over Genkin et al. (WO 2020/212756, publication date: 10/22/2020) in view of Qian et al. (WO 2019/020088, publication date: 01/31/2019), as applied to claims 1-5, 9, 10, 12, and 13, and further in view of CN112225822 (Publication date: 01/15/2021) and Leong et al. (US PG PUB 2020/0131244, publication date: 04/30/2020). As indicated above one of ordinary skill in the art would have been motivated with a reasonable expectation of success at the effective filing date of the invention to combine the teachings of Genkin et al. and Qian et al. to develop a CAR comprising a MSLN single-chain antibody (the instant SEQ ID NO: 8), a spacer domain or hinge region (CD8), a transmembrane region (CD28), an intracellular co-stimulatory domain (CD28), and a signal region (CD3ζ), linked in sequence. As indicated above Qian et al. teach CARs that target mesothelin, see throughout, and at [0032], Qian et al. teach a CAR that comprises a CD8 signal peptide, an anti-MSLN single chain antibody, a CD8 hinge region, a CD8 transmembrane region, a CD28 intracellular domain, and an intracellular activation domain from CD3ζ. At [0061], Qian et al. specifically teach a hinge region from CD8α. At [0016], Qian et al. teach the CD8 signal peptide of SEQ ID NO: 1, which comprises the instant SEQ ID NO: 2 (CD8α signal peptide). At [0061], Qian et al. teach the CD8α hinge of SEQ ID NO: 5, which comprises the instant SEQ ID NO: 14 (CD8α hinge). Qian et al. do not teach or suggest that said CAR comprises a CD8α signal peptide comprising SEQ ID NO: 2, a CD8α hinge region comprising SEQ ID NO: 14, a CD28 transmembrane region comprising SEQ ID NO: 4, a CD28 co-stimulatory domain comprising SEQ ID NO: 6, and a CD3ζ signal region comprising SEQ ID NO: 16. These deficiencies are remedied by CN112225822 and Leong et al. (US PG PUB 2020/0131244, publication date: 04/30/2020). At p. 9, CN112225822 teaches that “[t]he invention comprises a nucleic acid expression vector containing one or more elements and used for constructing the chimeric antigen receptor protein expressed on the surface of T cells, NK cells or NKT cells. Various commercially available vectors can be selected as needed, or the vectors can be constructed according to the conventional techniques in the field of molecular biology. In a specific embodiment, the vector used in the present invention is a lentiviral plasmid vector pLV300. This plasmid belongs to a fourth generation self-inactivating lentiviral vector system that has a total of four plasmids, namely a packaging plasmid encoding the protein Gag/Pol, a packaging plasmid encoding the Rev protein, an envelope plasmid encoding the VSV-G protein, and an empty vector pLV300, which can be used for the recombinant introduction of a nucleic acid sequence of interest, i.e. a nucleic acid sequence encoding a chimeric antigen receptor protein.” At p. 7, CN112225822 teaches a CAR construct comprising the CD28 co-stimulatory domain of SEQ ID NO: 23, which shares 100% sequence homology with the instant SEQ ID NO: 6, and a CD3ζ signal region comprising SEQ ID NO: 35, which shares 100% sequence homology with the instant SEQ ID NO: 16. At [0149], Leong et al. teach a CAR construct comprising the CD28 transmembrane region of SEQ ID NO: 4, which shares 100% sequence homology with the instant SEQ ID NO: 4. Based upon these teachings, one of ordinary skill in the art would have been motivated with a reasonable expectation of success at the effective filing date of the invention to prepare a CAR that comprises a CD8α signal peptide comprising SEQ ID NO. 2, a CD8α hinge region comprising SEQ ID NO. 14, a CD28 transmembrane region comprising SEQ ID NO: 4, a CD28 co-stimulatory domain comprising SEQ ID NO: 6, and a CD3ζ signal region comprising SEQ ID NO: 16. One of ordinary skill in the art would have been motivated to do so, because Qian et al. teach an anti-MSLN CAR for use in the treatment of MSLN-expressing cancers, wherein said CAR comprises a CD8α signal peptide comprising SEQ ID NO: 2 and a CD8α hinge region comprising SEQ ID NO: 14. Furthermore at p. 7, CN112225822 teaches a CAR construct comprising the CD28 co-stimulatory domain of SEQ ID NO: 23, which shares 100% sequence homology with the instant SEQ ID NO: 6, and a CD3ζ signal region comprising SEQ ID NO: 35, which shares 100% sequence homology with the instant SEQ ID NO: 16. Additionally at [0149], Leong et al. teach a CAR construct comprising the CD28 transmembrane region of SEQ ID NO: 4, which shares 100% sequence homology with the instant SEQ ID NO: 4. Therefore one of ordinary skill in the art would have been motivated to prepare an anti-MSLN CAR that comprises a CD8α signal peptide comprising SEQ ID NO: 2, a CD8α hinge region comprising SEQ ID NO: 14, a CD28 transmembrane region comprising SEQ ID NO: 4, a CD28 co-stimulatory domain comprising SEQ ID NO: 6, and a CD3ζ signal region comprising SEQ ID NO: 16, because said CAR could be used in the treatment of MSLN-expressing cancers. The invention of Qian et al., CN112225822, and Leong et al. meets the limitations of claim 6. Claim 7 is included in this rejection, because at [0039], Qian et al. teach expression vectors that comprise nucleic acids which can be used to express CARs. As such one of ordinary skill in the art would have been motivated to prepare nucleic acids that encode the anti-MSLN CAR of Qian et al., CN112225822, and Leong et al., thus arriving at the nucleotide sequences of the instant claim 7. Allowable Subject Matter Claims 8, 9, and 26 are objected to as being dependent upon a rejected base claim, but would be allowable if rewritten in independent form including all of the limitations of the base claim and any intervening claims. Conclusion No claims are allowed. Any inquiry concerning this communication or earlier communications from the examiner should be directed to NELSON B MOSELEY II whose telephone number is (571)272-6221. The examiner can normally be reached on M-F, 9:00-6:00 EST. 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If you would like assistance from a USPTO Customer Service Representative or access to the automated information system, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000. /NELSON B MOSELEY II/Primary Examiner, Art Unit 1642