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
1. A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 9/30/25 has been entered.
Applicant’s response filed 9/30/25 is acknowledged and has been entered.
2. Applicant is reminded of Applicant's election of Group I and species of genetic modification that is a CAR and expansion condition without bisphosphonate in Applicant’s response filed 11/29/24.
Claims 1-3 and 8-19 are presently being examined as they read upon the elected species as well as upon the species of non-genetically CAR modified cell population.
3. 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.
4. For the purpose of prior art rejections, the filing date of the instant claims 17 and 19 is deemed to be the filing date of PCT/EP2019/070125, i.e., 7/25/19, as provisional application 62/703,654 does not support the claimed limitations of the instant application. The said provisional application does not provide support for the activation condition and expansion conditions being performed simultaneously (claim 17) or tor the liver tissue, bone marrow, epithelial tissue, or cord blood samples recited in claim 19.
5. 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.
6. Claims 1-3, 8-14 and 17-19 stand rejected under 35 U.S.C. 103 as being unpatentable over Siegers and Lamb (Molecular Therapy, 2014, 22(8): 1416-1422, of record) in view of Schumm et al (Cytotherapy, 2013, 15: 1253-1258, of record), Lopez et al (Blood, 2000, 96(2): 3827-3837, of record), Miltenyi Biotech (2015, 3 pages, of record), and Dunne et al (J. Immunol. 2001, 167: 3129-3138, of record), as evidenced by Miltenyi Biotech CliniMACs TCR a/b Product Line (2021, 3 pages, of record).
Claim interpretation: The specification discloses that the term “about” as used herein refers to a value within 10% of the underlying parameter (i.e., plus or minus 10%), and the use of the term “about” at the beginning of a string of values modifies each of the values (i.e., about 1, 2, and 3 refers to about 1, about 2 and about 3) (page 107 at lines 26-29). The specification discloses that the term “depleted” as used herein, means that substantially all of the depleted component such as alpha.beta T cells, has been removed from the sample, for example at least about 80% (page 27 at lines 5-11). The specification does not disclose a limiting definition for the term “substantially”. In addition, the recitation of the open transitional phrase “comprising” or “comprises” opens the claims to encompass additional non-recited steps and/or ingredients.
Siegers and Lamb teach that both NK cells and gamma.delta T cells play a role in mediating antitumor immunity and that NK cells have been used for cancer immunotherapy. Siegers and Lamb teach that while NK cells can be easily isolated, expanded and activated ex vivo, Lopez was the first to develop a strategy of pan gamma.delta T cell expansion taking advantage of a CD2-initiated signaling pathway (citing Lopez’ use of an anti-CD2 antibody, see below) which resulted in a gamma.delta T cell population exhibiting inhibition of apoptosis while retaining potent innate antitumor activity against a wide variety of human hematopoietic and solid primary tumors and cell lines. Siegers and Lamb teach that this method is also advantageous in that it expands peripheral blood gamma.delta T cells regardless of phenotype and is adaptable to clinical scale use. Siegers and Lamb further teach that gamma.delta T cells can migrate into tumors and play a role in the prevention of both ALL and AML relapse. Siegers and Lamb teach that gamma.delta T cells can play an important role in cancer therapy as they are highly cytotoxic to epithelial and hematopoietic malignancies and have the added advantage of persistence over time. Siegers and Lamb teach that gamma.delta T cells also have anti-viral properties against CMV and EBV infection, both of which have been associated with malignant transformation. Siegers and Lamb teach that there is an immunomagnetic system for depletion of alpha .beta T cells from peripheral blood apheresis products that will allow investigators to infuse grafts enriched for gamma.delta T cells or as alpha.beta T cell depleted donor leukocyte infusions. Siegers and Lamb teach using a feeder cell free system for GMP manufacturing of clinical products (see entire reference, especially concluding remarks section, page 1419 at lines 2-11, page 1420 at column 1, the first sentence of the first full paragraph, abstract).
Thus, Siegers and Lamb teach myriad advantages of activating gamma.delta T cells by signaling through CD2 (using an anti-CD2 antibody) for production of gamma delta T cells for use in the clinic, for example, for treatment of a wide variety of human tumors or for treatment of virally-infected cells. These advantages include that the gamma.delta T cells retain potent innate anti-tumor activity against a wide range of tumors while resisting apoptosis (programmed self-destruction), the method activates peripheral blood gamma.delta T cells regardless of phenotype and is adaptable to clinical scale use, and the cells can migrate into tumors, and also teach depletion of alpha.beta T cells while retaining gamma.delta T cells (and inherently teaches also retaining NK cells and monocytes since peripheral blood contains NK cells and monocytes and since Siegers and Lamb also teach NK cells are a primary mediator of antitumor immunity).
Siegers and Lamb do not teach wherein the depleted cell population is exposed to at least one antibody or antigen-binding fragment thereof that immunospecifically binds to NKp46 besides stimulating through CD2 using an antibody specific for CD2, nor exposing the depleted cell population to expansion conditions comprising IL-15, nor wherein an antibody that immunospecifically binds to an alpha.beta TCR is the reagent that is used to deplete alpha.beta T cells from the peripheral blood apheresis products as is taught by the primary art reference.
Lopez et al teach isolating PBMCs (i.e., peripheral blood mononuclear cells), then culturing them feeder-free in the presence of a soluble agonistic [i.e., activating] anti-CD2 antibody (e.g., “S5.2”, mouse anti-human CD2) followed by stimulation with rhIL-2, resulting in first activation and then expansion of gamma.delta T cells. Lopez et al teach that accessory cells such as monocytes must be present to cross-link the anti-CD2 mAb(s), or in the absence thereof, the anti-CD2 mAb(s) must be immobilized. Lopez et al teach that these gamma.delta T cells can be readily expanded to sufficient numbers for use in various forms of adoptive immunotherapy, including treating tumors and viral infections. Lopez et al teach culturing for periods including three weeks. Lopez et al teach that these gamma.delta T cells are capable of mediating significant anti-tumor cytotoxicity against a panel of human derived tumor cell lines, but the prototypic NK-sensitive target cell K562 is relatively resistant to gamma.delta T cell mediated killing. Lopez et al exemplify activating the gamma.delta T cells for one day, then expanding the activated gamma.delta T cells for fourteen days or for twenty-one days (see entire reference, especially materials and methods, section spanning pages 3829-3830, Fig. 2, 3 legends).
Thus, Lopez et al also teach isolating PBMCs, and activating them in the presence of anti-CD2 antibody, followed by expansion of the resulting gamma.delta T cells. Lopez also teach that these gamma.delta T cells are capable of mediating significant anti-tumor cytotoxicity to many tumor cell lines, but not to the prototypic NK sensitive target cell line K562. The latter teaching indicates that anti-CD2 activated and later expanded gamma.delta T cells will not kill all types of tumor cells, including one that is susceptible to NK cell killing, and thus provide a reinforcement of the teaching of the primary art reference for using both gamma.delta T cells and NK cells to kill tumor cells and providing motivation to do so.
Miltenyi Biotech (2015) teaches that NK cells from PBMCs can be activated using a combination of anti-CD2 and anti-NKp46 (i.e., anti-CD355) antibodies immobilized on beads and expanded using IL-2 and in the presence of human serum and expansion for fourteen to eighteen days, but also teach that NK expansion is donor-dependent. Miltenyi Biotec further teaches that NK cells can kill the malignant K562 target cell line (that is resistant to killing by gamma.delta T cells). Miltenyi Biotech teaches simultaneous activation and expansion of the NK cells (see entire reference).
Thus Miltenyi Biotec (2015) teaches that NK cells from PBMCs are activated by anti-CD2 (like gamma.delta cells) in combination with anti-NKp46, a receptor present on NK cells but not on gamma.delta T cells, and that these activated NK cells can be expanded after activation and used to kill malignant cells.
Schumm et al teach that CliniMACsTM device can be used for depletion of alpha.beta T cell receptor and also CD19 positive (i.e., B cells) from apheresis products, with recovery of NK cells, gamma/delta positive T cells and mononuclear cells, advantageously using a single device for depletion of a large number of cells in an acceptable amount of time. Schumm et al teach that after depletion using the said CliniMACsTM device, a median of 0.00097% (0.00025-0.0048%) of TCRab+ T cells could be detected (results section at the first paragraph) (see entire reference).
Evidentiary reference Miltenyi Biotech CliniMACsTM TCR a/b Product Line (2021) teaches that the CliniMACsTM system uses an anti-TCR a/b antibody that recognizes a common determinant of the a/b TCR, and is the system used by Schumm et al and also that the CD19+ B cell depletion reagent (CliniMACsTM CD19 Reagent) was also used by Schumm et al (see entire reference, especially product overview, and reference 1 under “Publications”).
Dunne et al teach that IL-2 and IL-15 induce gamma.delta T cell expansion as well as NK cell expansion, while also stimulating cytotoxicity by NK cells against K562 targets. (see entire reference, especially abstract). Dunne et al exemplify about 76.7% of NK cells (within the range of “about” 80% provided by the definition of “about” in the instant specification).
It would have been prima facie obvious to one of ordinary skill in the art before the filing date of the claimed invention to have depleted alpha.beta T cells from PBMC (peripheral blood mononuclear cells from a peripheral blood sample) or an apheresis sample as is taught by the primary art reference using the anti-ab TCR antibody CliniMACsTM system taught by Schumm et al, subsequently activating the gamma.delta T cells and the NK cells using an agonistic anti-CD2 antibody(s) as taught by Siegers and Lamb and also by Lopez et al and by Miltenyi Biotech, and also using an anti-NKp46 antibody as taught by Miltenyi Biotech, and expanding the cells in IL-2 (taught by Lopez et al, Miltenyi Biotech, and Dunne et al) and additionally in IL-15 as taught by Dunne et al, for about the number of days required for expansion, using the teachings of the art references (i.e., for about fourteen to twenty-one days) as guidance for optimization of individual donor-specific gamma.delta and NK cell expansion, optimization being well-within the purview of one of ordinary skill in the art to determine.
One of ordinary skill in the art would have been motivated to do this to prepare compositions suitable for treatment of tumors and/or for study of investigational treatment of cancers.
Schumm et al teach depletion of these said alpha.beta T cells from an apheresis product with a median of 0.00097% alpha/beta TCR positive cells remaining in the apheresis product after depletion. In addition, it would have been prima facie obvious to one of ordinary skill in the art before the filing date of the claimed invention to have essentially removed the alpha.beta T cells. One of ordinary skill in the art would have been motivated to do this in order to prepare a composition that would cause minimal risk in an allogeneic setting, particularly in light of the teaching of the primary art reference.
Instant claim 12 is included in this rejection because Lopez et al teach that soluble anti-CD2 can activate gd T cells if monocytes are present, as is the case in the instant rejection (wherein ab T cells are depleted from PBMCs that inherently comprise monocytes) and because it would have been prima facie obvious to have also used a soluble anti-NKp46 antibody (particularly if a soluble anti-CD2 antibody were also to be used) because the expectation is that the antibody would be cross-linked through the constant region on monocytes as well. Instant claim 12 is included in this rejection because it would have been prima facie obvious to one of ordinary skill in the art to have used a soluble anti-CD2 antibody and anti-NKp46 antibody because monocytes having Fc receptors for cross-linking are present in the depleted cell population.
Claim 18 is also included in this rejection because Miltenyi Biotech teaches simultaneous activation and expansion of the NK cells (e.g., section “2.3” on page 2/3), and so it would have been prima facie obvious to institute both simultaneously for convenience sake.
Instant claims 13 and 14 are included in this rejection because the method of expanding the cells taught by the combined references is free of feeder cells and also free from bisphosphonate.
With regard to the limitation recited in instant base claim 1 “thereby generating the composition comprising about 80-99% NK cells”, it would have been prima facie obvious to one of ordinary skill in the art before the filing date of the claimed invention to have routinely optimized the expansion to achieve high levels of NK cells after expansion. In addition, the claim recites method steps and ingredients that are taught by the prior art references that result in such a recited percentage of NK cells.
Applicant’s arguments have been fully considered but are not persuasive.
Applicant’s said arguments are of record in the response filed 9/30/25 on pages 3-11.
Applicant’s said arguments are largely the same as those previously submitted. The Examiner’s response to Applicant’s prior said arguments appears below in smaller text following the Examiner’s rebuttal of Applicant’s newly presented arguments.
Applicant newly argues that Siegers (i.e., Siegers and Lamb) does not primarily focus on NK cell enrichment. However, this argument is not persuasive because Applicant is arguing the references separately. Siegers teaches that both NK cells and gamma.delta T cells play a role in mediating antitumor immunity, that NK cells have been used for cancer immunotherapy, that there is a GMP-compliant immunomagnetic system for depletion of alpha .beta T cells from peripheral blood apheresis products that will allow investigators to infuse grafts enriched for gamma.delta T cells or as alpha.beta T cell depleted donor leukocyte infusions, and a GMP-compliant method for using anti-CD2 to expand gamma.delta T cells.
Applicant further argues that Schumm only describes that a specific device can be used for depletion of ab T cell receptor [positive cells] without specifying the residual amount of ab T cells in the sample. However, Schumm et al teach that after depletion using the said CliniMACsTM device, a median of 0.00097% (0.00025-0.0048%) of TCRab+ T cells could be detected. Even if this were not the case, as is enunciated in the claim interpretation section above, the instant specification discloses that the term “depleted” as used herein, means that substantially all of the depleted component such as alpha.beta T cells, has been removed from the sample, for example at least about 80% (page 27 at lines 5-11). The specification does not disclose a limiting definition for the term “substantially”. The art reference Schumm teaches depletion of alpha.beta T cells. it would have been prima facie obvious to one of ordinary skill in the art before the filing date of the claimed invention to have essentially removed the alpha.beta T cells. Additionally, as is stated in the instant rejection, one of ordinary skill in the art would have been motivated to deplete alpha.beta T cells in order to prepare a composition that would cause minimal risk in an allogeneic setting, particularly in light of the teaching of the primary art reference.
Applicant further argues that Dunne is silent with regard to the choice of the polypeptide in the expansion condition, that choice being essential to obtain the desirable amount of NK cells, and in particular when the polypeptide is IL-15, the resulting population of cells enriched in NK cells often contains about 80-99% NK cells and about 1-20% gd T cells. However, Applicant mischaracterizes Dunne, as Dunne et al teach that IL-2 and IL-15 induce gamma.delta T cell expansion as well as NK cell expansion, while also stimulating cytotoxicity by NK cells against K562 targets. Dunne et al exemplify about 76.7% of NK cells (within the range of “about” 80% provided by the definition of “about” in the instant specification).
Applicant argues the alleged differences between Siegers alone and the instant invention, citing the description in Siegers of the cytotoxic and regulatory properties of NK cells as tumor immunotherapeutics versus the claimed invention in which the NK cells are further enriched (a first step of depleting ab T cells that generates a depleted cell population comprising NK cells and gd cells. Applicant argues that this step is of particular relevance, since in the composition of the present invention the ab T cells are absent or present in negligible to low amounts. Applicant argues that Siegers on the other hand, only describes that there is an immunomagnetic system for depletion of ab T cells from peripheral blood apheresis products without specifying the residual amount of ab T cells. However, Siegers and Lamb teach the first step that depletes ab T cells, and Schumm et al teach that after depletion using the same system, a median of 0.00097% (0.00025-0.0048%) of TCRab+ T cells could be detected. In addition as is
enunciated in the instant rejection, it would have been prima facie obvious to one of ordinary skill in the art before the filing date of the claimed invention to have essentially removed the alpha.beta T cells in order to prepare a composition that would cause minimal risk in an allogeneic setting, particularly in light of the teaching of the primary art reference Siegers and Lamb. The cited references in the instant rejection provide teachings and motivation to combine the references to expand both NK cells and gamma.delta T cells. Thus Applicant is arguing the references separately. In response to Applicant's arguments against the references individually, one cannot show nonobviousness by attacking references individually where the rejections are based on combinations of references. See In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981); In re Merck & Co., 800 F.2d 1091, 231 USPQ 375 (Fed. Cir. 1986).
Applicant further argues that starting from the teachings of Siegers, the skilled person would not have been motivated to choose a first polypeptide that immunospecifically binds to CD2 and a second polypeptide that immunospecifically binds to NKp46 for the activation step. However Siegers and Lamb teach that anti-CD2 expands gamma.delta T cells, while Miltenyi Biotech (2015) teaches that NK cells from PBMCs can be activated using a combination of anti-CD2 and anti-NKp46 (i.e., anti-CD2 expands gamma.delta T cells as well as NK cells, while anti-NKp46 [the other specificity in combination with anti-CD2] working through an NK cell specific receptor stimulates NK cells].
Applicant also argues that the composition for immunotherapy that is made up of innate immune cells such as NK cells are advantageous over those containing adaptive immune cells. However, Applicant’s argument is off-point as the instant rejection teaches depleting adaptive immune cells (i.e., the alpha.beta T cells); it does not purport to retain them.
Applicant argues that the choice of polypeptide in the expansion condition is essential to obtain the desirable amount of NK cells; for example, when the polypeptide is IL-15, the resulting population of cells enriched in NK cells often contains about 80-99% of NK cells and about 1-20% gamma.delta T cells, and the composition finds greater applicability against solid tumors. Applicant also argues that Miltenyi Biotech 2015 is silent with regard to the type of IL-15 or the percentage of NK cells in the composition that can be obtained using IL-15, and therefore staring from Milteny Biotech 2015, the skilled person would not have been motivated to select IL-15 to generate [the recited composition]. Applicant further argues that Dunne cannot provide the missing link to the combination of references because it is silent with regard to the choice of IL-15 that is essential to obtain the desirable amount of NK cells. However, the instant rejection adds IL-15 and Applicant is arguing Miltenyi Biotech 2015 separately. It bears repeating that Dunne et al teach that IL-2 and IL-15 induce gamma.delta T cell expansion as well as NK cell expansion, while also stimulating cytotoxicity by NK cells against K562 targets [i.e., the targets that gamma.delta cells do not kill]. Dunne et al exemplify about 76.7% of NK cells (within the range of “about” 80% provided by the definition of “about” in the instant specification). Instant base claim 1 recites “thereby generating the composition comprising about 80-99% NK cells” after recitation of all of the active method steps therein. The instant rejection performs all of the recited method steps, thereby generating the composition comprising about 80-99% NK cells. The instant rejection performs all of the active method steps, thereby generating a composition comprising about 80-99% NK cells.
Applicant also alleges that the need to combine a greater number of references is evidence of non-obviousness. However, In response to Applicant's argument that the Examiner has combined an excessive number of references, reliance on a large number of references in a rejection does not, without more, weigh against the obviousness of the claimed invention. See In re Gorman, 933 F.2d 982, 18 USPQ2d 1885 (Fed. Cir. 1991).
The Examiners response to Applicant’s prior arguments of record appear immediately below.
Applicant argues that the primary art reference Siegers (and Lamb) only provides for the potential of Vd1+gd T cells in cancer immunotherapy, that they can be cytotoxic against tumors or exhibit immunosuppressive/regulatory properties, persist longer in circulation and are less prone to activation-induced cell death, but is silent with regard to NK cell enrichment.
However Siegers and Lamb also teach that both NK cells and gd T cells play a role in mediating antitumor immunity, that NK cells can be easily isolated, expanded, and activated ex vivo, gd T cells can be activated by signaling through CD2 (using an anti-CD2 antibody) after peripheral blood products (comprising NK cells and monocytes) are depleted of ab T cells in a GMP compliant fashion for clinical therapeutic application in the clinic.
Applicant further argues that while NK cells are often mentioned in the broader context of immune cell therapies, the art reference focuses more on the application of the gd T cells rather than T cells. Applicant states that said reference is silent with regard to exposing the depleted cell population to at least one antibody or antigen-binding fragment thereof that immunospecifically binds to NKp46 besides stimulating through CD2 using an antibody specific for CD2 as well as exposing the abT cell-depleted cell population to expansion comprising use of IL-15, wherein the depletion uses an antibody that immunospecifically binds to an ab TCR.
However, Siegers and Lamb clearly teach that NK cells can be easily expanded and activated ex vivo for the same clinical application as gd T cells for treating tumors in the clinic. In addition, Lopez et al teach that the prototypic NK-sensitive tumor target cell K562 is resistant to gd T cell-mediating killing, an indication that gd T cells will not kill all types of tumors cells, as well as teaching activation of gd T cells for anti-tumor therapy using feeder-free conditions, anti-CD2 antibody, and recombinant human IL-2, and conditions therefore. Applicant is also arguing Siegers and Lamb separately. In response to Applicant's arguments against the references individually, one cannot show nonobviousness by attacking references individually where the rejections are based on combinations of references. See In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981); In re Merck & Co., 800 F.2d 1091, 231 USPQ 375 (Fed. Cir. 1986).
In arguing that Schumm fails to disclose a method for obtaining a composition comprising about 80-99% NK cells and about 1% to about 20% gd T cells, Applicant is also arguing the references separately, as the depletion step is only a portion of the instantly recited method. Applicant argues that Schumm teaches 80% recovery of NK cells, 83% recovery of gd T cells, and 79% recovery of CD14+ cells (i.e., a marker for monocytes, the cells that art reference Lopez et al teach as necessary to cross-link the anti-CD2 antibodies). However, Table II of Schum indicates that the said recovery rates pertain only to the amount of the particular said cell type before ab T cell depletion versus after. In contrast the instant claimed method recites the final composition range of about 80-99% NK cells. Applicant also argues Lopez et al as well as Miltenyi Biotech 2015 separately in arguing that said reference is silent with regard to a method for enriching populations in NK cells or in teaching a composition with the recited % of NK and gd T cells, respectively. Applicant is also arguing the references separately when arguing that Dunn fails to disclose the depleting ab T cells step and exposing it to anti-CD2 as well as the final cell percentages (after all the recited steps in the claimed method). Applicant states that (evidentiary reference) Miltenyi 2021 only describes an antibody-anti- ab TCR antibody that allows for ab T cell depletion while retaining NK and gd T cells, while being silent with regard to all the features of the presently claimed method.
Applicant alleges that for all of the forgoing reasons, the combination of the cited references would not have rendered obvious the presently claimed invention; none of the cited references, alone or in combination, describe all the claimed limitations, and even if combined they would not suggest or disclose a method for NK cell enrichment as presently claimed; a person of ordinary skill in the art would find no suggestion or motivation to combine and modify the cited references t arrive at the presently claimed subject matter. Applicant argues that the mere identification in the art of various elements of the claims is not sufficient, per se, to support the conclusion of obviousness.
In response to Applicant’s argument that there is no teaching, suggestion, or motivation to combine the references, the Examiner recognizes that obviousness may be established by combining or modifying the teachings of the prior art to produce the claimed invention where there is some teaching, suggestion, or motivation to do so found either in the references themselves or in the knowledge generally available to one of ordinary skill in the art. See In re Fine, 837 F.2d 1071, 5 USPQ2d 1596 (Fed. Cir. 1988), In re Jones, 958 F.2d 347, 21 USPQ2d 1941 (Fed. Cir. 1992), and KSR International Co. v. Teleflex, Inc., 550 U.S. 398, 82 USPQ2d 1385 (2007). In the instant case, Applicant is mischaracterizing the instant rejection. The references cited therein teach the importance of expanding and activating both NK cells and gd cells for tumor therapy in the clinic, ways of activating both cell types after depletion of ab T cells (both cell types as well as monocytes are present in the depleted cell population in a GMP compliant fashion, the monocytes being necessary for cross-linking anti-CD2 antibodies), and that some tumor targets that are NK-cell sensitive are not sensitive to killing by gd T cells, an additional teaching of the usefulness of both in said cell types in treating tumors as well as motivation for activating and expanding both said cell types. As the art teaches the same method steps using the same ingredients as are recited in the presently claimed invention, the resulting percentages of NK cells and gd cells must necessarily inherently be the same.
7. Claims 15 and 16 stand rejected under 35 U.S.C. 103 as being unpatentable over Siegers and Lamb (Molecular Therapy, 2014, 22(8): 1416-1422, of record) in view of Schumm et al (Cytotherapy, 2013, 15: 1253-1258, of record), Lopez et al (Blood, 2000, 96(2): 3827-3837, of record), Miltenyi Biotech (2015, 3 pages, of record), and Dunne et al (J. Immunol. 2001, 167: 3129-3138, of record), as evidenced by Miltenyi Biotech CliniMACs TCR a/b Product Line (2021, 3 pages, of record) as applied to claims 1-3, 8-14 and 17-19 above, and further in view of Fisher and Anderson (Front. Immunol. 6/26/2018, 9, Article 1409: 1-10, of record), Boisssel et al (Leuk. Lymphoma, 2012, 53(5): 958-965, of record), and Capsomidis et al (Molec. Ther., 2/2018, 262(2): 354-365, of record).
Claim interpretation: The specification discloses that the term “about” as used herein refers to a value within 10% of the underlying parameter (i.e., plus or minus 10%), and the use of the term “about” at the beginning of a string of values modifies each of the values (i.e., about 1, 2, and 3 refers to about 1, about 2 and about 3) (page 107 at lines 26-29). The specification discloses that the term “depleted” as used herein, means that substantially all of the depleted component such as alpha.beta T cells, has been removed from the sample, for example at least about 80% (page 27 at lines 5-11). The specification does not disclose a limiting definition for the term “substantially”. In addition, the recitation of the open transitional phrase “comprising” or “comprises” opens the claims to encompass additional non-recited steps and/or ingredients.
The teachings and combination of Siegers and Lamb (in view of Schumm et al (Cytotherapy, 2013, 15: 1253-1258), Lopez et al (Blood, 2000, 96(2): 3827-3837, IDS reference), Miltenyi Biotech (2015 ), and Dunne et al (J. Immunol. 2001, 167: 3129-3138), as evidenced by Miltenyi Biotech CliniMACs TCR a/b Product Line has been enunciated above, hereafter referred to as “the combined references.”
The combined references do not teach that the method further comprises subjecting the depleted cell population to a genetic modification condition after exposing the depleted cell population to an expansion condition (as is recited in dependent claim 15), nor wherein the genetic modification comprises introducing an exogenous polynucleotide encoding a chimeric antigen receptor (CAR) into the cells (as is recited in dependent claim 16).
Fisher and Anderson teach engineering NK cells and gamma.delta T cells with chimeric antigen receptors (CARs) and their use in treating tumors. Fisher and Anderson teach that gamma.delta T cells retain graft-versus-leukemia activity in the hematopoietic stem cell transplant setting (see entire reference, especially sections on NK cells and gamma.delta T cells).
Boisssel et al teach engineering NK cells with chimeric antigen receptors (CARs) recognizing CD19 or CD20 on tumor cells in order to facilitate killing of resistant B-lymphoid cells and primary B-CLL (B cell chronic lymphocytic leukemia cells), and using these cells in clinical trials (see entire reference).
Capsomidis et al teach genetically engineering human gamma.delta T cells because such cells are primed for rapid cytotoxicity toward cancer cells, can function as professional APCs following activation, and when engineered with a CAR, work by focusing T cell function on defined cell surface tumor antigens and provide essential costimulation for robust activation
It would have been prima facie obvious to one of ordinary skill in the art before the filing date of the claimed invention to have engineered the activated or activated and expanded NK and gamma.delta T cell population with an exogenous polynucleotide construct encoding a CAR.
One of ordinary skill in the art would have been motivated to do this in order to reap the advantages of CAR-engineering of NK cells and gamma.delta T cells taught by the secondary references above.
Applicant’s said arguments are of record in the response filed 9/30/25 on page 11. Applicant argues that claim 1 is patentable over the combination of references cited above at item #6 of this office action for the reasons above, and claims 15 and 16 are as well.
Applicant’s arguments have been fully considered but are not persuasive for the reasons cited above at item #6 of this office action..
8. Claims 1-3, 8-14 and 17-19 stand rejected under 35 U.S.C. 103 as being unpatentable over Siegers and Lamb (Molecular Therapy, 2014, 22(8): 1416-1422, of record) in view of Schumm et al (Cytotherapy, 2013, 15: 1253-1258, of record), Lopez et al (Blood, 2000, 96(2): 3827-3837, of record), Miltenyi Biotech (2015, 3 pages, of record), Koepsell et al (Transfusion, 2013, 53: 404-410, of record), and Van Acker et al (J. Hematol. & Oncol. 2016, 9:101: 1-13, of record), as evidenced by Miltenyi Biotech CliniMACs TCR a/b Product Line (2021, 3 pages, of record).
Claim interpretation: The specification discloses that the term “about” as used herein refers to a value within 10% of the underlying parameter (i.e., plus or minus 10%), and the use of the term “about” at the beginning of a string of values modifies each of the values (i.e., about 1, 2, and 3 refers to about 1, about 2 and about 3) (page 107 at lines 26-29). The specification discloses that the term “depleted” as used herein, means that substantially all of the depleted component such as alpha.beta T cells, has been removed from the sample, for example at least about 80% (page 27 at lines 5-11). The specification does not disclose a limiting definition for the term “substantially”. In addition, the recitation of the open transitional phrase “comprising” or “comprises” opens the claims to encompass additional non-recited steps and/or ingredients.
Siegers and Lamb teach that both NK cells and gamma.delta T cells play a role in mediating antitumor immunity and that NK cells have been used for cancer immunotherapy. Siegers and Lamb teach that while NK cells can be easily isolated, expanded and activated ex vivo, Lopez was the first to develop a strategy of pan gamma.delta T cell expansion taking advantage of a CD2-initiated signaling pathway (citing Lopez’ use of an anti-CD2 antibody, see below) which resulted in a gamma.delta T cell population exhibiting inhibition of apoptosis while retaining potent innate antitumor activity against a wide variety of human hematopoietic and solid primary tumors and cell lines. Siegers and Lamb teach that this method is also advantageous in that it expands peripheral blood gamma.delta T cells regardless of phenotype and is adaptable to clinical scale use. Siegers and Lamb further teach that gamma.delta T cells can migrate into tumors and play a role in the prevention of both ALL and AML relapse. Siegers and Lamb teach that gamma.delta T cells can play an important role in cancer therapy as they are highly cytotoxic to epithelial and hematopoietic malignancies and have the added advantage of persistence over time. Siegers and Lamb teach that gamma.delta T cells also have anti-viral properties against CMV and EBV infection, both of which have been associated with malignant transformation. Siegers and Lamb teach that there is an immunomagnetic system for depletion of alpha .beta T cells from peripheral blood apheresis products that will allow investigators to infuse grafts enriched for gamma.delta T cells or as alpha.beta T cell depleted donor leukocyte infusions. Siegers and Lamb teach using a feeder cell free system for GMP manufacturing of clinical products (see entire reference, especially concluding remarks section, page 1419 at lines 2-11, page 1420 at column 1, the first sentence of the first full paragraph, abstract).
Thus, Siegers and Lamb teach myriad advantages of activating gamma.delta T cells by signaling through CD2 (using an anti-CD2 antibody) for production of gamma delta T cells for use in the clinic, for example, for treatment of a wide variety of human tumors or for treatment of virally-infected cells. These advantages include that the gamma.delta T cells retain potent innate anti-tumor activity against a wide range of tumors while resisting apoptosis (programmed self-destruction), the method activates peripheral blood gamma.delta T cells regardless of phenotype and is adaptable to clinical scale use, and the cells can migrate into tumors, and also teach depletion of alpha.beta T cells while retaining gamma.delta T cells (and inherently teaches also retaining NK cells and monocytes since peripheral blood contains NK cells and monocytes and since Siegers and Lamb also teach NK cells are a primary mediator of antitumor immunity).
Siegers and Lamb do not teach wherein the depleted cell population is exposed to at least one antibody or antigen-binding fragment thereof that immunospecifically binds to NKp46 besides stimulating through CD2 using an antibody specific for CD2, nor exposing the depleted cell population to expansion conditions comprising IL-15, nor wherein an antibody that immunospecifically binds to an alpha.beta TCR is the reagent that is used to deplete alpha.beta T cells from the peripheral blood apheresis products.
Lopez et al teach isolating PBMCs (i.e., peripheral blood mononuclear cells), then culturing them feeder-free in the presence of a soluble agonistic [i.e., activating] anti-CD2 antibody (e.g., “S5.2”, mouse anti-human CD2) followed by stimulation with rhIL-2, resulting in first activation and then expansion of gamma.delta T cells. Lopez et al teach that accessory cells such as monocytes must be present to cross-link the anti-CD2 mAb(s), or in the absence thereof, the anti-CD2 mAb(s) must be immobilized. Lopez et al teach that these gamma.delta T cells can be readily expanded to sufficient numbers for use in various forms of adoptive immunotherapy, including treating tumors and viral infections. Lopez et al teach culturing for periods including three weeks. Lopez et al teach that these gamma.delta T cells are capable of mediating significant anti-tumor cytotoxicity against a panel of human derived tumor cell lines, but the prototypic NK-sensitive target cell K562 is relatively resistant to gamma.delta T cell mediated killing. Lopez et al exemplify activating the gamma.delta T cells for one day, then expanding the activated gamma.delta T cells for fourteen days or for twenty-one days (see entire reference, especially materials and methods, section spanning pages 3829-3830, Fig. 2, 3 legends).
Thus, Lopez et al also teach isolating PBMCs, and activating them in the presence of anti-CD2 antibody, followed by expansion of the resulting gamma.delta T cells. Lopez also teach that these gamma.delta T cells are capable of mediating significant anti-tumor cytotoxicity to many tumor cell lines, but not to the prototypic NK sensitive target cell line K562. The latter teaching indicates that anti-CD2 activated and later expanded gamma.delta T cells will not kill all types of tumor cells, including one that is susceptible to NK cell killing, and a motivation and reinforcement of the teaching of the primary art reference for using both gamma.delta T cells and NK cells to kill tumor cells.
Miltenyi Biotech (2015) teaches that NK cells from PBMCs can be activated using a combination of anti-CD2 and anti-NKp46 (i.e., anti-CD355) antibodies immobilized on beads and expanded using IL-2 and in the presence of human serum and expansion for fourteen to eighteen days, but also teach that NK expansion is donor-dependent. Miltenyi Biotec further teaches that NK cells can kill the malignant K562 target cell line (that is resistant to killing by gamma.delta T cells). Miltenyi Biotech teaches simultaneous activation and expansion of the NK cells (see entire reference).
Thus Miltenyi Biotec (2015) teaches that NK cells from PBMCs are activated by anti-CD2 (like gamma.delta cells) in combination with anti-NKp46, a receptor present on NK cells but not on gamma.delta T cells, and that these activated NK cells can be expanded after activation and used to kill malignant cells.
Schumm et al teach that CliniMACsTM device can be used for depletion of alpha.beta T cell receptor and also CD19 positive (i.e., B cells) from apheresis products, with recovery of NK cells, gamma/delta positive T cells and mononuclear cells, advantageously using a single device for depletion of a large number of cells in an acceptable amount of time. Schumm et al teach that after depletion using the said CliniMACsTM device, a median of 0.00097% (0.00025-0.0048%) of TCRab+ T cells could be detected (results section at the first paragraph) (see entire reference).
Evidentiary reference Miltenyi Biotech CliniMACsTM TCR a/b Product Line (2021) teaches that the CliniMACsTM system uses an anti-TCR a/b antibody that recognizes a common determinant of the a/b TCR, and is the system used by Schumm et al and also that the CD19+ B cell depletion reagent (CliniMACsTM CD19 Reagent) was also used by Schumm et al (see entire reference, especially product overview, and reference 1 under “Publications”).
Koepsell et al teach that it is advantageous to include IL-15 in the expansion conditions over use of IL-2 alone to improve viability and potency of NK cells. Koepsell et al also teach that it is desirable to deplete B cells. Koepsell et al teach that B cells have been implicated in the development of passenger lymphocyte syndrome after the infusion of NK cell products, including wherein the donor B cells had EBV virus reactivation with subsequent hemophagocytic reaction resulting in death. Koepsell et al teach that given these complications and the fact that B cells do not contribute to NK cell expansion, survival or cytolytic activity, B cell depletion of NK cell products is important for providing a safe cellular product. (See entire reference, especially page 407 at column 1, para 1, and page 405 at column 2, para 3).
Van Acker et al teach that addition of IL-15 to gamma.delta T cell cultures (including from PBMCs) results in a more activated phenotype, a higher proliferative capacity, a more pronounced Th1 polarization and increased cytotoxic capacity and anti-tumor properties of the gamma.delta T cells. Van Acker et al teach that IL-2 can be included in the expansion (see entire reference, especially abstract and page 3 at column 1, last para).
It would have been prima facie obvious to one of ordinary skill in the art before the filing date of the claimed invention to have depleted alpha.beta T cells from PBMC (peripheral blood mononuclear cells from a peripheral blood sample) or an apheresis sample as is taught by the primary art reference using the anti-ab TCR antibody CliniMACsTM system taught by Schumm et al, subsequently activating the gamma.delta T cells and the NK cells using an agonistic anti-CD2 antibody(s) as taught by Siegers and Lamb and also by Lopez et al and by Miltenyi Biotech, and also using an anti-NKp46 antibody as taught by Miltenyi Biotech, and expanding the cells in IL-2 (taught by Lopez et al, Miltenyi Biotech, and Dunne et al) and additionally in IL-15 as taught by Koepsell et al and Van Acker et al, for about the number of days required for expansion, using the teachings of the art references (i.e., for about fourteen to twenty-one days) as guidance for optimization of individual donor-specific gamma.delta and NK cell expansion, optimization being well-within the purview of one of ordinary skill in the art to determine.
One of ordinary skill in the art would have been motivated to do this to prepare compositions suitable for treatment of tumors and/or for study of investigational treatment of cancers.
Schumm et al teach depletion of these said alpha.beta T cells from an apheresis product with a median of 0.00097% alpha/beta TCR positive cells remaining in the apheresis product after depletion. In addition, it would have been prima facie obvious to one of ordinary skill in the art before the filing date of the claimed invention to have essentially removed the alpha.beta T cells. One of ordinary skill in the art would have been motivated to do this in order to prepare a composition that would cause minimal risk in an allogeneic setting, particularly in light of the teaching of the primary art reference.
Instant claim 12 is included in this rejection because Lopez et al teach that soluble anti-CD2 can activate gd T cells if monocytes are present, as is the case in the instant rejection (wherein ab T cells are depleted from PBMCs that inherently comprise monocytes) and because it would have been prima facie obvious to have also used a soluble anti-NKp46 antibody (particularly if a soluble anti-CD2 antibody were also to be used) because the expectation is that the antibody would be cross-linked through the constant region on monocytes as well. Instant claim 12 is included in this rejection because it would have been prima facie obvious to one of ordinary skill in the art to have used a soluble anti-CD2 antibody and anti-NKp46 antibody because monocytes having Fc receptors for cross-linking are present in the depleted cell population.
Claim 18 is also included in this rejection because Miltenyi Biotech teaches simultaneous activation and expansion of the NK cells (e.g., section “2.3” on page 2/3), and so it would have been prima facie obvious to institute both simultaneously for convenience sake.
Instant claims 13 and 14 are included in this rejection because the method of expanding the cells taught by the combined references is free of feeder cells and also free from bisphosphonate.
With regard to the limitation recited in instant base claim 1 “thereby generating the composition comprising about 80-99% NK cells”, it would have been prima facie obvious to one of ordinary skill in the art before the filing date of the claimed invention to have routinely optimized the expansion to achieve high levels of NK cells after expansion. In addition, the claim recites method steps and ingredients that are taught by the prior art references that result in such a recited percentage of NK cells.
Applicant’s said arguments are of record in the response filed 9/30/25 on pages 11-12. Applicant states that the presently claimed invention, Siegers, Schumm, Lopez, Miltenyi 2015 [as evidenced by] Miltenyi 2021 have been discussed above. Applicant further argues that both Koepsell and Van Acker fail to teach all the features presently claimed.
and are based upon the allegation that neither Koepsell nor Van Acker disclose all the features of claim 1, the addition of their teachings [to the other cited references therein] do not correct the deficiencies of the combination of the cited references above.
Applicant’s arguments have been fully considered but are not persuasive for the reasons cited above at item #6 of this office action.
9. Claims 15 and 16 stand rejected under 35 U.S.C. 103 as being unpatentable over Siegers and Lamb (Molecular Therapy, 2014, 22(8): 1416-1422, of record) in view of Schumm et al (Cytotherapy, 2013, 15: 1253-1258, of record), Lopez et al (Blood, 2000, 96(2): 3827-3837, of record), Miltenyi Biotech (2015, 3 pages, of record), Koepsell et al (Transfusion, 2013, 53: 404-410, of record), and Van Acker et al (J. Hematol. & Oncol. 2016, 9:101: 1-13, of record), as evidenced by Miltenyi Biotech CliniMACs TCR a/b Product Line (2021, 3 pages, of record) as applied to claims 1-3, 8-14 and 17-19 above, and further in view of Boisssel et al (Leuk. Lymphoma, 2012, 53(5)