DETAILED CORRESPONDENCE
Applicant’s amendment and response received on 5/5/26 has been entered. Claims 1-20 are currently pending and under examination in the instant application. The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA . An action on the merits follows.
Those sections of Title 35, US code, not included in this action can be found in a previous office action.
Claim Rejections - 35 USC § 102
The rejection of claims 1-11 under 35 U.S.C. 102(a)(1) OR under 35 U.S.C. 102(a)(2) as being anticipated by WO 2015/142661, published on 9/24/15, with an effective filing date of 3/15/14, and hereafter referred to as Engels et al., is withdrawn in view of applicant’s amendments to the claims which now require that the CAR comprises an Nkp30 antigen binding domain.
The rejection of claims 1-11 under 35 U.S.C. 102(a)(2) as being anticipated by US Patent Application Publication 2015/0376296 (2015), with an effective filing date of 3/15/13, and hereafter referred to as Fedorov et al., is withdrawn in view of applicant’s amendments to the claims which now require that the CAR comprises an Nkp30 antigen binding domain.
Claim Rejections - 35 USC § 112
Applicant’s amendments to the claims has necessitated the following new or modified grounds of rejection set forth below.
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.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
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 of carrying out his invention.
Claim 2 is newly 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. This is a new matter rejection.
The applicant is reminded that an amendment to the claims or the addition of a new claim must be supported by the description of the invention in the application as filed. In re Wright, 866 F.2d 422, 9 USPQ2d 1649 (Fed. Cir. 1989). New or amended claims which introduce elements or limitations which are not supported by the as-filed disclosure violate the written description requirement. See, e.g., In re Lukach, 442 F.2d 967, 169 USPQ 795 (CCPA 1971); In re Smith, 458 F.2d 1389, 1395, 173 USPQ 679, 683 (CCPA 1972).
Claim 2 depends on claim 1 and has been amended to recite, “wherein the antigen binding domain or a antigen binding fragment thereof binds to B7h”. Claim 1 has been amended to recite an Nkp30 antigen binding domain or antigen binding fragment thereof. The as filed specification does not disclose “B7h” or teach that an NKp30 antigen binding domain can bid to “B7h”. It is noted that the specification does disclose B7-H6, which is the ligand for Nkp30. Thus, while the specification provides written description for B7-H6 as a ligand for the Nkp30, the specification provides no description of B7h and does not teach than any such “B7h” can be bound by Nkp30. As such, the recitation that the NKp30 antigen binding domain binds to B7h represents new matter not disclosed by the specification.
The rejection of amended claims 1-20 under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, for scope of enablement, is maintained in modified form as follows. The previous rejection was a scope of enablement rejection; however, in view of applicant’s amendments to the claims, the rejection has been modified as follows: claims 1-20 are now rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the enablement requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to enable one skilled in the art to which it pertains, or with which it is most nearly connected, to make and/or use the invention. Applicant’s amendments to the claims and arguments have been fully considered but have not been found persuasive in overcoming the modified rejection for reasons of record as discussed in detail below.
The applicant has amended independent claim 1 to recite a chimeric antigen receptor (CAR) comprising three polypeptides, a first polypeptide comprising an Nkp30 antigen binding domain or an antigen binding fragment thereof, a second polypeptide comprising transmembrane and cytoplasmic domains of CTLA-4, and a third polypeptide comprising the cytoplasmic domain of CD3zeta. Dependent claim 4 as amended now recites that the CAR further comprises a fourth polypeptide derived from an intracellular domain of an inhibitory coreceptor selected from a group consisting of LAG-3, PD-1, TIM-3, TIGIT, BTLA, LILRB4, LILB3, CD160, 2B4, LAIR-1, CD66a, CD44, and neuropilin-1 (NRp1). Independent claim 12 has been amended to recite a CAR comprising three polypeptides, an antigen targeting domain which binds to an antigen or ligand at a site of inflammation or autoimmunity, and intracellular signaling domains of CTLA-4, and an intracellular domain of NRp1. Claims 9 and 17 recite methods of treating chronic inflammation or immune-mediated autoimmunity by delivering to a subject in need an effective amount of FoxP3+T cells or Treg cells modified to express the CAR of claim 1 or claim 12 respectively.
35 U.S.C. 112(a) requires that the specification provide sufficient description 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 invention as claimed. The specification identifies that the use of the CAR is for expression in a Treg cell where binding of the CAR to its ligand results in a Treg cell with therapeutic activity against inflammation or autoimmunity. The specification further identifies that the use of the Treg cell is for administration in vivo to treat a chronic inflammatory disease or autoimmune disease. In regards to the CAR of claim 1, it is noted that the CAR as currently claimed comprises both an activating domain which is the intracellular CD3zeta signaling domain and an inhibitory domain which is the intracellular CTLA4 signaling domain. The specification fails to provide an enabling disclosure that such a CAR whose intracellular region comprises both an activating domain which is the intracellular CD3zeta signaling domain and an inhibitory domain which is the intracellular CTLA4 signaling domain is capable of activating a Treg cell in response to the binding of the CAR to its ligand on a target cell, and in particular to activate the Treg cell such that it has a therapeutic effect in a subject with a chronic inflammatory disease or an autoimmune disease. The specification discloses a CAR comprising a co-inhibitory CTLA-4 intracellular domain, and further discloses that the extracellular domain of the CAR can be an Nkp30 antigen binding domain/target antigen binding domain which binds to a target antigen/ligand associated with inflammation or autoimmunity. In particular, the specification teaches that the target antigen is B7H6 which is expressed in salivary tissue. The specification provides a single working example which describes making a nucleic acid construct encoding a CAR comprising a first polypeptide comprising an Nkp30 antigen binding domain or an antigen binding fragment thereof, a second polypeptide comprising transmembrane and cytoplasmic domains of CTLA-4, and a third polypeptide comprising the cytoplasmic domain of CD3zeta. The working example states that the CAR was expressed in the packaging cell line E86 and predicts that the CAR would also be expressed on the cell surface of transduced Treg cells. However, the working example does not actually transduce any T cell or Treg cell with the nucleic acid construct encoding the CAR and further does not provide any data regarding the ability of this CAR to activate Treg cells when bound to a target ligand which binds to Nkp30, such as B7H6. The working examples do not provide any description of the functional activity of this CAR when expressed from any cell, including a Treg cell. The working examples further does not demonstrate any therapeutic activity for the CAR when expressed from a Foxp3+ T cell or Treg cell, or demonstrate any therapeutic activity for any T cells expressing this CAR in any subject suffering from any inflammatory or autoimmune disease.
In regards to the putative activity of the co-inhibitory intracellular CTLA-4 domain in CAR, the specification teaches that it has been shown that CTLA-4 engagement can enhance Foxp3 induction in naïve CD4+ T cells, citing Barnes et al. (2013) Mucosal Immunol., Vl. 6(2), 324-334. The specification speculates that including a co-inhibitory signaling domain of CTTLA-4 in a CAR could activate an immune cell toward an immune suppressive phenotype (specification, pages 13-14). However, Barnes et al., cited by the specification, was concerned with the generation of Treg cells from naïve CD4+ T cells. Barnes et al. was interested in the role of CTLA4 is the generation of FoxP3+ Treg cells form naïve CD4+ T cells in outside the thymus/in the periphery. Barnes et al. shows through the of CTLA4 knockout naïve CD4 Tc cells while signaling through CTLA4 on the cell surface of CD4 cells is not necessary for the development of Treg cells and the expression of Foxp3 both in vitro and in vivo (Barnes et al., page 325). Barnes does show that in contrast to Treg induction in the thymus, and in the spleen and mesenteric lymph nodes, CTLA4 does appear to promote increased number of Foxp3 CD4 T cells in the colon lamia propria (Barnes et al., page 325). However, Barnes et al. was concerned with the effects of signaling through surface expressed CTLA-4 on the generation of Foxp3 Treg cells in naïve T cells before they become Treg cells. Barnes et al. is silent on the effect of CTLA-4 signaling in Foxp3 Treg cells which have already been generated, or in Foxp3 Treg cells which have been activated by antigen. Barnes et al. is also silent as the effect of signals from an intracellular domain of CTLA-4 present in a CAR on Treg function. At the time of filing, treatment of inflammatory or autoimmune disease using Treg expressing a CAR utilized activating CAR comprising an extracellular antigen binding domain that bound to an autoantigen, a transmembrane region, and activating intracellular signaling domains such as the intracellular CD3zeta domain or Fc-chain receptor chain (first generation CAR), or the intracellular CD3zeta domain in combination with co-stimulatory domain such as the intracellular CD28 domain (second generation CAR). Elinav et al., for example, teaches Foxp3+ Treg cells transduced with vector encoding CAR comprising an antibody specific for TNP, an antigen associated with experimentally induced TNBS colitis, fused to the extracellular and transmembrane domain of CD28 and the intracellular domain of the stimulatory Fc-gamma receptor chain (Elinav et al. (2009) Gastroenterology, Vol. 136(5), 1721-1731, page 1721). Elinav et al. teaches that the CAR-Treg cells were delivered to mice with acute TNBS colitis resulting in 60% improvement in survival (Elinav et al., page 1724). Likewise, Blat et al. teaches Foxp3+ Treg cells transduced with a CAR comprising an extracellular antibody domain that binds to carcinoembryonic antigen (CEA), a transmembrane domain and intracellular CD28 and CD3zeta signaling domains (Blat et al. (2014) Mol. Ther., Vol. 22(5) 1018-1028, page 1026). Blat et al. teaches that CEA is an inflammatory marker for inflamed colons and teaches that administration of the anti-CEA CAR Treg to a cell transfer model of colitis, where mice had previously developed colitis as result of administration of CEA specific CAR CD4+ T effector (Teff) cells, significantly inhibited abdominal accumulation and expansion of the Teff cells resulting in substantially reduced severity of colitis and increased survival (Blat et al., pages 1021-1023). Thus, at the time of filing, CAR-Treg therapy utilized CAR comprising only activating intracellular domains. Turning to the state of the art at the time of filing concerning CAR with inhibitory intracellular domains, the prior art teaches that CAR comprising inhibitory domains were called inhibitory CAR (iCAR), and contained an antigen-binding domain, a transmembrane domain, and an inhibitory signaling domain such as the intracellular domain of CTLA4, PD-1, LAG-3, or BTLA (Fedorov et al., of record, paragraphs 30-32). Fedorov et al. teaches that the CTLA4 intracellular signaling domain present in the iCAR sends intracellular signals that are immunosuppressive and which inhibit T-cell function upon antigen recognition (Fedorov et al., paragraphs 30-32). Fedorov et al. teaches that iCAR can be used in combination with activating CAR in T cells to improve target selectivity, where the activating CAR recognizes a disease antigen present on the target cell, and the iCAR recognizes an antigen which is present on normal or non-target cells (Fedorov et al., Figure 1). Fedorov further teaches and demonstrates both in vitro and in vivo that in cells which express both the target disease antigen recognized by the CAR and the normal or non-target antigen recognized by the iCAR, signaling through the iCAR inhibits the signaling through the activating CAR in the T cell, thus preventing activation of the T cell (Fedorov et al., and paragraphs 85-89). Thus, the prior art of record shows that signaling through the intracellular CTLA4 domain inhibits the effects of signaling through intracellular activating domains such as CD3zeta. As such, the skilled artisan at the time of filing would not have predicted that a CAR comprising both a CD3zeta intracellular signaling domain and a CTLA4 intracellular signaling domain when expressed by a Treg cell would be capable of activating the Treg cell, or that any such CAR Treg cells would be capable of being activated in vivo in order to treat any inflammatory or autoimmune disease.
It is noted that applicant’s arguments addressing previously raised issues regarding ITIM domains is not found persuasive as the claims as amended have not been rejected for lack of an ITIM domain.
In regards to claim 4, which further recites the presence of an additional “co-inhibitory” intracellular domain selected from Lymphocyte- Activation Gene 3 (LAG-3), Programmed cell death protein 1(PD-1), T cell Immunoglobulin Mucin-3 (TIM-3), T-cell immunoreceptor with immunoglobulin (TIGIT), B-and T-lymphocyte Attenuator (BTLA), leukocyte immunoglobulin-like receptor subfamily B member 4 (LILRB4), LILRB3, CD160, 2B4, Leukocyte-Associated Immunoglobulin-like Receptor 1 (LAIR-1), CD66a, CD44, or neuropilin-1 (NRp1), it is noted that the specification does not disclose the specific activity of any Nkp30 CAR comprising three intracellular domains which are CD3zeta, CTL4, and one of the additional “co-inhibitory” intracellular domains listed above. As discussed in detail above, the prior art at the time demonstrated that in T cells comprising a CAR and iCAR comprising the intracellular domain of CTLA-4, signaling through CTLA-4 inhibited signaling and activation through CD3zeta. The prior art of record further teaches, as also discussed above, that CAR-Treg therapeutic activity requires an activating signal leading to Treg cell activation. Thus, the skilled artisan at the time of filing, would not have predicted that the inclusion of an additional “co-inhibitory” signaling domain would lead to Treg activation. Furthermore, as discussed in the rejection of record, the specification, while identifying 2B4, CD160, CD44, and Npr1 as co-inhibitory receptors, fails to provide an enabling disclosure for any signaling domains derived from any of 2B4, CD160, CD44, or Npr1 which when incorporated into a CAR molecule are capable of transducing an inhibitory/co-inhibitory signal into a T cell, Treg cell, or any other immune cell. In regards to CD44, CD44 is not taught by the specification to include either an ITIM or ITAM motif. In reference to CD44, the specification provides a single paragraph disclosing that CD44 is a cell adhesion molecule which is the major hyaluronan receptor, and has been implicated in the binding, endocytosis, and metabolism of hyaluronan (HA) (specification, paragraph 36). Paragraph 36 further cites Teder et al. (2002) Science, Vol. 296, 155-158, for teaching that in bleomycin-induced acute lung injury, CD44-deficient mice show an enhanced and persistent inflammatory response due to impaired clearance of apoptotic neutrophils and HA fragments from the injury site. Paragraph 36 of the specification also cites Kawana, et al. (2008) J. Immunol. Vol.180, 4235-45, for showing that CD44 directly associates with TLR2 when stimulated by the TLR2 ligand zymosan and that the cytoplasmic domain of CD44 is crucial for its regulatory effect on TLR signaling. In addition, paragraph 36 states that CD44 negatively regulates in vivo inflammation mediated by Toll-Like Receptors (TLRs) via NF-.kappa.B activation, which leads to proinflammatory cytokine production. The specification also discloses alleged CD44 signaling domains provided as GENBANK accession numbers (specification, paragraph 36); however, it is noted that these accession numbers are to 8 full length CD44 isoforms, and do not identify any “signaling” domain(s).
The specification, while alluding to the use of a CD44 signaling domain, does not disclose any specific signaling domain from CD44, or any specific functional activity for any domain within the cytoplasmic domain of CD44. Turning to the references cited by the specification, Teder et al., cited in the specification, teaches that CD44 is important in clearing HA (nonsulfated glycosaminoglycan hyaluronan) which accumulated at sites of inflammation and tissue injury (Teder et al., page 155). Teder et al. demonstrates bleomycin induced lung injury and inflammation associated with increases in HA is increased in CD44 deficient mice, and that administration of bone marrow expressing wild type CD44 reduces both HA and inflammation in this mouse model (Teder et al., page 156-158). CD44 binding and clearance of HA is a function of the full length CD44 molecule. Teder et al. does not teach that the cytoplasmic domain of CD44 has any specific anti-inflammatory role in reducing inflammation in this mouse model, and further does not teach or suggest that the cytoplasmic domain of CD44 can be used as a signaling domain in a chimeric receptor to affect the effector or regulatory function of T cells or other immune cells in which is expressed, particularly for the purpose of treating any inflammatory or autoimmune disease. Kawana et al., also cited by the specification, actually teaches that a considerable number of publications have reported that CD44 expression plays a crucial role in a variety of inflammatory diseases and that CD44 expression is upregulated on a number of inflammatory cells in these diseases (Kawana et al., page 4235). Kawana et al. teaches that inhibiting CD44 can inhibit inflammation in arthritis, cutaneous inflammation, experimental autoimmune encephalomyelitis, and IL-2 induced vascular leak syndrome (Kawana et al., page 4235). Kawana et al. does acknowledge the teachings of Teder regarding the role of CD44 in reducing HA and thus inflammation in bleomycin-induced acute lung injury, and further demonstrates that CD44 can negatively regulate TLR2 signaling in zymosan induced inflammation, an effect that appears to require the a full length CD44 molecule including the cytoplasmic domain, (Kawana et al., pages 4235, and 4242-4244. However, Kawana et al. does not teach that the CD44 cytoplasmic domain, by itself or when part of fusion protein, can be used to transduce signals directly or indirectly in any type of immune cell, or teach the effects of any such putative CD44 mediated signals on the effector or regulatory functions of immune cells such as T cells, B cells, dendritic cells, macrophages, or eosinophils. In fact, Kawana et al. states that it is evident that the function of CD44 in inflammation is complex and involves multiple cells types, ligands, and signaling pathways (Kawana et al., page 4236). Other publications from the prior art support this variable role of CD44 in various cell types. Baatan et al., for example, teaches that CD44 expression on T cells has been shown to be upregulated on naïve T lymphocytes following exposure to microbes, and that the relevance of CD44 expression to T-cell responses or homeostasis has been largely unexplored (Baatan et al. (2010) Communicative & Integrative Biology, Vol. 3(6), 508-512, see page 508). Baatan et al. does teach that CD44 is involved in the regulation of CD4 T cell survival, but not other T cell subpopulations, and further teaches that in Treg cells, CD44 is associated with the expression of FoxP3 and the cytokines TGFb1 and IL-10 (Baatan et al., page 509). However, again, Baatan et al. does not teach any specific signaling domains present in the cytoplasmic region of CD44 or show that any portion of the intracellular region of CD44 can mediate any particular type of signal in any type of T cell. In fact, Baatan et al. explicitly states that CD44 lacks intrinsic signaling activity and the signaling pathways coupled to CD44 are not fully defined (Baatan et al., page 510). Thus, the prior art at the time of filing establishes that the state of the art for the role of CD44 in inflammation and autoimmunity, and the role of CD44 in immune cells, including T cells and Treg cells, was largely undefined and unpredictable, with CD44 expression linked to both inducing/increasing inflammation in some models of inflammatory and autoimmune disease, and in reducing inflammation in other models. The prior art also establishes the unpredictability at the time of filing for identifying and using a “signaling” domain from CD44 in immune cells due to both the art-recognized lack of intrinsic signaling activity by the CD44 molecule, and the fact that the intracellular pathways by which CD44 mediates its pleiotropic effects on various cells had not been fully elucidated. Further, the prior art, like the specification, does not provide specific guidance for any “signaling” domain present in the intracellular region of CD44 and further does not teach how any putative “signaling” sequence derived from CD44 can be used in a chimeric protein such as a chimeric antigen receptor to transduce any type of signal within any immune cell, including and T cell or Treg cell as claimed. The state of the prior art as discussed above highlights the undeveloped and unpredictable nature of using any putative “signaling” domain from CD44 within a CAR by itself, or with any of signaling domain form any one or more of the co-inhibitor receptor recited in the claims, as the effects of any CD44 sequence on the functionality of such as CAR could not have been predicted a priori. Applicant’s arguments do not address this issue.
Likewise for 2B4 and CD160, while the specification states that both of these molecules are co-inhibitory molecules, the specification fails to provide sufficient guidance for any specific signaling domain from 2B4 or CD160 with any specific inhibitory activity in any immune cell including a T cell or Treg cell. At the time of filing, Lee et al. teaches that 2B4 is an unusual NK receptor which does not contain either an ITIM or ITAM domain, but which has stimulatory activity in cells (Lee et al. (2004) J. Exp. Med., Vol. 199(9), 1245-1254, see page 1245). Lee et al. does teach that while 2B4 has an immunoreceptor tyrosine-based switch motif (ITSM) in the cytoplasmic domain, the ITSM domains are stimulatory domains (Lee et al., page 1255). Lee et al. teaches that 2B4 interacts with intracellular SH2D1A, and that mutations in SH2D1A can result in an inhibitory signal from 2B4. However, neither Lee et al., nor the instant specification, teach a signaling domain from 2B4 which is inherently inhibitory or under which conditions a signaling domain from 2B4 may be inhibitory in any and all immune cells. Turning to CD160, the art at the time of filing teaches that the main isoform of CD160 is in fact a GPI anchored protein, which has an single IgV-like extracellular domain and a GPI anchor (del Rio et al. (2010) J. Leuk. Biol., Vol. 87, 223-235, see page 231). Thus, the GPI isoform of CD160 does not in fact have any cytoplasmic domain and neither the prior art nor the specification identify any co-inhibitory domain derived from the IgV-like extracellular domain or the GPI anchor. Giustiniani et al. further teaches that while a second isoform of CD160 with a short cytoplasmic tail has been identified, the signals transduced through this molecule are activating signals, not inhibitory signals (Giustiniani et al. (2009) J. Immunol., Vol. 182, 63-71, see page 63). Neither the prior art nor the instant specification teach a signaling domain obtained form any isoform of CD160 which is inherently inhibitory or under which conditions a signaling domain from CD160 may be inhibitory in any and all immune cells. Again, applicant’s arguments do not address this issue.
In regards to amended claim 12 and a CAR comprising three polypeptides which are an antigen targeting domain which binds to an antigen or ligand at a site of inflammation or autoimmunity, and intracellular signaling domains of CTLA-4, and an intracellular domain of NRp1, the analysis of the teachings of the specification and the working examples discussed in detail above apply. In particular, note that the specification working examples do not demonstrate the functional effects of any CAR comprising an intracellular signaling domain of CTLA-4, or a combination of the intracellular domains of CTLA-4 and Nrp1, nor do the working examples describe or demonstrate the functional effects of any such CAR in a Treg cell or demonstrate that Treg expressing a CAR which has an extracellular antigen targeting domain which binds to an antigen or ligand at a site of inflammation or autoimmunity, and intracellular signaling domains of CTLA-4, and an intracellular domain of NRp1 is capable of ameliorating or treating any inflammatory condition or autoimmune disease. Further, as set forth in the extended discussion of the state of the prior art at the time of filing, the prior art only teaches the use of an iCAR comprising a CTLA-4 intracellular domain in a T cell in combination with an activating CAR, where the iCAR functions to suppress the activation of the dual CAR/iCAR T cell on off-target cells when the iCAR is bound to its non-target antigen. Neither the specification nor the prior art teaches that a Treg which expresses only an iCAR comprising inhibitory intracellular domains can have any therapeutic effect on any inflammatory condition or autoimmune disease.
Further in regards to the addition of an intracellular domain of Nrp1 to an iCAR comprising he intracellular domain of CTLA4, the rejection of record stated that the specification discloses that neuropilin (Nrp1) is not characterized as a co-inhibitory receptor (specification, page 23). The specification states that Nrp1 expression can suppress autoreactive T cells in an experimental autoimmune encephalomyelitis model, citing Solomon et al. (2001) PNAS, Vol. 108, 2040-2045, and further states that gene-expression analysis shows that Nrp-1 induced transcriptional profile is consistent with the promotion of T regulatory cell survival, citing Delgoffe et al. (2013) Nature, Vol. 501, 252-256. The specification also discloses alleged Nrp1 signaling domains provided as GENBANK accession numbers (specification, paragraph 37); however, it is noted that these accession numbers are to full length Nrp1 genes derived from a number of mammals. The specification does not disclose any specific signaling domain from Nrp-1, or any specific functional activity for any domain within the Nrp-1 protein and specifically the cytoplasmic domain of Nrp-1. Turning to the references cited by the specification, it is noted that Solomon et al. looked at Npr-1 deficient Treg cells, and while they concluded that Nrp-1 expression is important for suppressing CD4+ autoreactive immune cells, Solomon et al. does not teach that Npr-1 participates in any particular signal transduction pathway or identify any portions or domains or Npr-1 which are necessary or responsible for signaling within a T cell. Delgoffe et al. mentions the potential importance of a three amino acid SEA motif at the C-terminus of Npr-1 in recruiting PTEN for potentiating Treg function and survival, but again does not teach any specific “signaling domain” within Npr-1 or teach which sequences in the Npr-1 protein, or intracellular region in particular, which are necessary and sufficient to transduce any signal in a T cell when present within a native Npr-1 protein or any type of chimeric protein.
The rejection of record also discussed that at the time of filing, the art teaches that Nrp-1 (neuropilin 1) is a pleiotropic glycoprotein which was first identified as an axonal adhesion protein, was later found to associate with both VEGF-A (VEGF165) and SEMA3A, and has a role in angiogenesis, arteriogenesis, cell migration, and cell adhesion (Plein et al. (2014) Microcirculation, Vol. 21, 315-323, page 316). Plein et al. teaches that Nrp-1 is a glycoprotein with a large extracellular domain responsible for binding VEGF165 and SEMA3A, a transmembrane domain, and a short cytoplasmic domain with no known catalytic activity (Plein et al., pages 316 and 318, and Figure 1). Plein et al. does teach that the cytoplasmic domain comprises a C-terminal SEA motif which can recruit synectin/GIPC1/NIP, a modulator of endocytic trafficking (Plein et al., page 316). Plein et al. postulates that since Nrp-1 lack any known catalytic activity that it transduces signals through coreceptors, such as VEGFR2 which associate with Nrp-1 through VEGF165 binding, and which activate intracellular signal transduction pathways involving ERK/MAPK, AKT1, SRC etc. (Plein et al., page 318). The Nrp-1 cytoplasmic domain SEA domain in this scenario binds to synectin/NIP and mediates endocytosis of the NRP-1 coreceptor complex (Plein et al., page 318). Takamatsu et al. further teaches that Nrp-1 bound to SEMA3A forms a complex with Plexin-A as a coreceptor in T cells, and postulates that signaling through the complex is transduced through the plexin component, with Nrp-1 serving to support or strengthen SEMA3A binding in the complex (Takamatsu et al. (2012) Trends in Immunology, Vol. 33(3), 127-135, see page 128 and Figure 1). Delgoffe et al., cited by the specification as noted above, teaches that SEMA4A also binds to Nrp1 and in T cells, specifically Treg cells, can recruit PTEN and restrain TCR activated Akt phosphorylation and signal transduction thus potentiating Treg function and survival (Delgoffe et al. (2013) Nature, Vol. 501(7466), 252-256, see page 252 and Supplemental Figure 15). According to Delgoffe et al., this Treg Npr-1 activity appears to only be important for Treg suppression of anti-tumor immune responses and inflammatory colitis, not autoimmunity (Delgoffe et al., page 252). While Delgoffe et al. focuses on demonstrating the binding of SEMA4A to Nrp-1, the prior art, such as Takamatsu et al. cited above, teaches that SEMA4A binds to a number of other receptors present on T cells, such as TIM-2 and various plexin-B subfamily members (Takamatsu et al., pages 128 and 131, and Figure 1). However, based on the teachings of Takamatsu et al. and Plein et al., that Nrp-1 transduces signals as part of a complex between Nrp-1, a ligand, and a coreceptor, it is unclear whether the Nrp-1-SEMA4A binding and subsequent signal transduction occur through the Nrp-1 protein itself or as part of larger complex with a plexin co-receptor on T cells. Further, the prior art of record is clear that the intracellular portion of Nrp-1 does not have any catalytic activity itself, and while the three amino acid SEA domain at the C-terminus has been implicated in recruitment of either NIP or PTEN, there are no teachings that such recruitment by itself results in any specific signaling versus signaling through co-receptors associated in complex with Nrp-1. Thus, the prior art does not provide specific guidance that the intracellular region of Nrp-1 has any “signaling” domain capable of transducing signals into a T cell, and further does not teach how any sequence derived from Nrp-1 can be used in a chimeric protein such as a chimeric antigen receptor to transduce any type of signal within a Treg cell as claimed. Thus, the teachings of the prior art of record demonstrate that the understanding of signal transduction following ligation of Nrp-1 and any of its known ligands in a T cell was in its infancy, with the identification and elucidation of Nrp-1 ligand binding, complex formation, co-receptor association, and intracellular signaling partners/signaling pathways having barely begun. The state of the prior art as discussed above highlights the undeveloped and unpredictable nature of using any putative “signaling” domain from Nrp-1 or even the entire short intracellular region of Nrp-1 within a CAR which also comprises a CTLA-4 signaling domain, , as the effects of any Nrp-1 sequence on the functionality of such as CAR could not have been predicted a priori.
Applicant’s argument that the amendment which now recites an “intracellular domain” instead of a “signaling domain” is enabled because the location of the intracellular region of Nrp1 was known is not found persuasive. As discussed in detail below, the prior art of record demonstrates that the understanding of signal transduction following ligation of Nrp-1 and any of its known ligands in a T cell was in its infancy, with the identification and elucidation of Nrp-1 ligand binding, complex formation, co-receptor association, and intracellular signaling partners/signaling pathways having barely begun. The state of the prior art as discussed above also highlights the undeveloped and unpredictable nature of using any putative “signaling” domain from Nrp-1 or even the entire short intracellular region of Nrp-1 within a CAR which also comprises a CTLA-4 signaling domain, as the effects of any Nrp-1 sequence on the functionality of such as CAR could not have been predicted a priori.
Therefore, based on state of the prior art at the time of filing which shows iCAR comprising an inhibitory intracellular domain such as CTLA-4 inhibit rather than activate T cells, and further that signaling through the intracellular CTLA4 domain of an iCAR inhibits the effects of signaling through intracellular activating domains such as CD3zeta, the teachings of the prior art that the therapeutic activity of Treg cells in treating inflammation/autoimmunity require activation of the Treg cells, not inhibition, the underdeveloped and unpredictable state of the prior art at the time of filing for using any intracellular signaling domain or intracellular domain derived from Nrp-1, CD44, 2B4, or CD160 as an inhibitory signaling domain in a CAR, particularly in the context of a CAR comprising additional signaling domains from other proteins, the lack of any working examples demonstrating the functional effects of a CAR comprising the intracellular domain of CTLA-4, the intracellular domains or both CTLA-4 and CD3zeta, or the intracellular domains or CTLA-4 and Nrp1 in a Treg cell, or the therapeutic effect from the administration of Treg cells expressing a CAR comprising the extracellular antigen binding domain that binds to an antigen or ligand at the site of inflammation or autoimmunity, and an intracellular domain of CTLA-4, the intracellular domains or both CTLA-4 and CD3zeta, or the intracellular domains or CTLA-4 and Nrp1 on any inflammatory disease or autoimmune disease, and the breadth of the claims, it would have required undue experimentation to make and use the invention as claimed.
No claims are allowed.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
Any inquiry concerning this communication from the examiner should be directed to Anne Marie S. Wehbé, Ph.D., whose telephone number is (571) 272-0737. If the examiner is not available, the examiner’s supervisor, Maria Leavitt, can be reached at (571) 272-1085. For all official communications, the technology center fax number is (571) 273-8300. Please note that all official communications and responses sent by fax must be directed to the technology center fax number. For informal, non-official communications only, the examiner’s direct fax number is (571) 273-0737. For any inquiry of a general nature, please call (571) 272-0547.
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Dr. A.M.S. Wehbé
/ANNE MARIE S WEHBE/Primary Examiner, Art Unit 1634
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