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 .
Claims 1, 3-4, 6, 9-11 and 14-19 are pending.
Claims 15-18 are withdrawn from further consideration by the examiner, 37 C.F.R. 1.142(b) as being drawn to non-elected inventions.
Claims 1, 3-4, 6, 9-11, 14 and 19, drawn to an isolated dimer comprising a first polypeptide and a second polypeptide, each polypeptide comprising a human IgG Fc region comprising a variant CH2 domain each variant CH2 domain comprising a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution, wherein the amino acid positions are numbered according to the EU index of Kabat, are being acted upon in this Office Action.
Priority
Applicant’ claim priority to provisional applications 61/845,948 filed July 12, 2013 and 61/671,640 filed July 13, 2012, is acknowledged.
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, or 365(c) is acknowledged. Applicant has not complied with one or more conditions for receiving the benefit of an earlier filing date under 35 U.S.C. 120 as follows:
The later-filed application must be an application for a patent for an invention which is also disclosed in the prior application (the parent or original nonprovisional application or provisional application). The disclosure of the invention in the parent application and in the later-filed application must be sufficient to comply with the requirements of the first paragraph of 35 U.S.C. 112. See Transco Products, Inc. v. Performance Contracting, Inc., 38 F.3d 551, 32 USPQ2d 1077 (Fed. Cir. 1994).
The disclosure of the prior-filed application 61/671,640 fails to provide adequate support or enablement in the manner provided by the first paragraph of 35 U.S.C. 112 for one or more claims of this application. The provisional application 61/671,640 does not disclose the particular substitutions L234A, L235A, D265S as recited in claim 1, T350V, L351Y, F405A and Y407V as recited in claim 6, T350V, T366L, K392L and T394W as recited in claim 6. Therefore for the purposes of applying prior art, the effective filing date of claims 1, 3, 6, 9-11, 14 and 19 is July 12, 2013, the date that provisional application 61/845,948 was filed.
The 61/671,640, 61/845,948, 13/941,449 disclose the heteromultimer is devoid of immunoglobulin light chain and immunoglobulin first constant (CH1) region, which corresponds to devoid of Fab since Fab comprises VH-CH1 and VL-CL. Therefore for the purposes of applying prior art, the effective filing date of claim 4 is January 24, 2019, the date that the 16/256,824 was filed.
Should applicant disagree with the examiner’s factual determination above, applicant should point to evidence that shows that the invention of claims 1, 3-4, 6, 9-11, 14 and 19 is in fact described in one or more of the previously-filed applications.
Information Disclosure Statement
The information disclosure statement (IDS) submitted December 30, 2025 has been considered by the examiner and an initialed copy of the IDS is included with this Office Action.
Specification
The substitute specification filed on December 30, 2025 has been entered.
Rejection withdrawn
The rejection of claims 1-4, 7, 8, 9, 10, 13 and 14 under 35 U.S.C. 102 (a)(1) as being anticipated by WO2004/099249 publication (published November 2004; PTO 1449) as evidenced by Hezareh (J Virology 75(24): 12161-12168, 2001; PTO 1449) is withdrawn in view of claim amendment.
The declaration of Genevieve Desjardins under 37 CFR 1.132 filed December 30, 2025 and applicant’s argument at p. 17 of the amendment filed December 30, 2025 are sufficient to overcome the rejection of claims 1 and 6 based upon 35 U.S.C. 103 as being unpatentable over W02004/099249 publication (of record, published November 2004; PTO 1449) as evidenced in Hezareh et al (J Virology 75(24): 12161-12168, 2001; PTO 1449) in view of US Pat No. 9,574,010 (claimed earliest priority to 61/556,090, filed Nov 4, 2011; PTO 1449).
The declaration of Genevieve Desjardins under 37 CFR 1.132 filed December 30, 2025 and applicant’s argument at p. 21 of the amendment filed December 30, 2025 are sufficient to overcome the rejection of claims 1 and 6 based upon 35 U.S.C. 103 as being unpatentable over US Pat No. 8,101,720 (of record, issued Jan 24, 2012; PTO 1449) in view of US Pat No. 9,574,010 (of record, claimed earliest priority to 61/556,090, filed Nov 4, 2011; PTO 1449) and Hezareh et al (of record, J Virology 75(24): 12161-12168, 2001; PTO 1449).
The rejection of claims 1, 5, 12, 19-20 and 21 under 35 U.S.C. 103 as being unpatentable over WO2004/099249 publication (published November 2004; PTO 1449) or US Pat No. 8,101,720 (issued Jan 24, 2012; PTO 1449) each in view of Von Kreudenstein et al (US20120149876 published June 14, 2012; PTO 892) and Bargou et al (Science 321: 974-977, August 15, 2008; PTO 14490) is withdrawn in view of the claim amendment.
Claim rejections under - 35 U.S.C. 112
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.
Claims 1, 3-4, 6, 9-11, 14 and 19 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for pre-AIA the inventor(s), at the time the application was filed, had possession of the claimed invention.
MPEP § 2163 states that the written description requirement for a claimed genus may be satisfied through sufficient description of a representative number of species by actual reduction to practice, or by disclosure of relevant, identifying characteristics, i.e., structure or other physical and/or chemical properties, by functional characteristics coupled with a known or disclosed correlation between function and structure, or by a combination of such identifying characteristics, sufficient to show the applicant was in possession of the claimed genus.
Claim 1 encompasses an antibody comprising a first polypeptide construct and a second polypeptide construct, each polypeptide construct comprising a human IgG Fc region sequence comprising a variant CH2 domain, each variant CH2 domain comprising a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution,
wherein the first polypeptide construct Fc region sequence and the second polypeptide construct Fc region sequence form a heterodimeric Fc region,
wherein one of the first and second polypeptide constructs further comprises a CD3 antigen-binding domain that binds to CD3, wherein the CD3 antigen-binding domain is a Fab or an scFv,
wherein the human IgG Fc region is an IgG1 Fc region, and
wherein the amino acid positions are numbered according to the EU index of Kabat.
Claim 3 encompasses the isolated dimer of claim 2, wherein the antigen-binding domain comprises an scFv.
Claim 4 encompasses the isolated dimer of claim 2, wherein the antigen-binding domain comprises a Fab.
Claim 6 encompasses the isolated dimer of claim 1, wherein one of the first and second polypeptide further comprises amino acid substitutions T350V, L351Y, F405A, and Y407V, and the other of the polypeptides further comprises amino acid substitutions T350V, T366L, K392L, and T394W.
Claim 9 encompasses the e antibody of claim 1, wherein the antibody is any monoclonal antibody, or any humanized antibody.
Claim 10 encompasses the antibody of claim 1, wherein the antibody is multispecific.
Claim 11 encompasses the antibody of claim 1, wherein the antibody is bispecific.
Claim 14 encompasses a pharmaceutical composition comprising the antibody of claim 1 and a pharmaceutically acceptable carrier.
Claim 19 encompasses the isolated dimer of claim 1, wherein each of the first and second polypeptide construct further comprises an antigen-binding domain, and wherein each antigen-binding domain binds a different antigen.
The specification discloses bispecific asymmetric heterodimer comprising having the following structure:
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It should be noted that the antigen-binding domain that binds to CD3 is from just anti-CD3 scFv called BITE.
The specification exemplifies:
[0359] v4541 has an anti-CD3 BiTE™ (VH-VL) 44-100SS scFv on chain A and anti-CD19 BiTE™ (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 50 and 52] Cyno/Human Cross-Reactive Anti-CD3 and Anti-CD19 Bispecific Fc Knock-Out Variants with or without Disulfide 44-100 Stabilization
[0360] v4542 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VH-VL) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 54 and 56].
[0361] v4543 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VH-VL) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 58 and 60]
[0362] v4544 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 62 and 64]
[0363] v4545 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VL-VH) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 66 and 68]
[0364] v4546 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VH-VL) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 70 and 72]
[0365] v4547 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VH-VL) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 74 and 76].
[0366] v4548 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 78 and 80] v4549 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VL-VH) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 82 and 84]
Regarding multispecific antibody, the specification discloses:
Example 3: Heteromultimer v873 is able to Bridge Jurkat CD3 T Cells and Raji CD19 B Cells
Example 4: Heteromultimers Bind Selectively to CD3- and CD19-Expressing Cells
[0384] The ability of an exemplary heteromultimer, v873, to bind specifically to CD3 and CD19 was assessed by FACS. One-armed antibodies (OAAs) against CD3 and CD19 were also prepared as described in Example 2 and tested as controls in the whole cell FACS binding assay described below.
Example 5: Heteromultimers Mediate PBMC Killing of Target Raji B Cells
Example 6: Heteromultimers Mediate Redirected Killing of Target Raji B Cells with Resting and IL-2 Activated CD4+ and CD8+ T Cells
[0405] The ability of an exemplary heteromultimers, v875, v1379, v1380 to mediate CD4+ and CD8+ T cell cytoxicity against target Raji B-cells was measured as described in Example 5.
Example 7: The Heterodimeric Fc Contributes to Target Raji B Cell Cytotoxicity
[0410] The ability of exemplary heteromultimers, v875, and v873, to mediate target Raji B-cell cytotoxicity in the presence and absence of Fc was measured as described in Example 5.
Example 8: Heteromultimers Mediate Autologous B Cell Cytotoxicity
[0413] The ability of exemplary the heteromultimer v875 and v873 to kill autologous B-cells was measured in total and resting IL-2-stimulated PBMCs where the percent of CD19+ 7AAD+ cells following incubation with v875 and v873 (300 nM, n=3 donors) was determined by flow cytometry as described in Example 5.
Example 9: Heteromultimer v875 Spares Autologous T Cell Cytotoxicity Compared to BiTE
[0415] The effects of exemplary heteromultimer v875 and v873 treatment on the autologous T cell population was assessed in total and resting IL-2-stimulated PBMCs where the percent of CD3+ 7AAD+ cells following incubation with v875 and v873 (300 nM, n=3 donors) was determined by flow cytometry as described in Example 5.
[0416] FIG. 22 shows, relative to untreated media and human IgG controls, v875 has a more selective B cell killing by sparing more autologous T cells compared to v873 and v891.
Example 10: Design, Expression and Purification of Heteromultimers with an Albumin Scaffold
[0417] The following exemplary CD3-CD19 binding heteromultimers based on an albumin scaffold were designed and prepared as follows.
[0418] The sequences for the anti-CD19 and anti-CD3 scFvs were chosen from two molecules that are currently in clinical trials and are well documented and tested for stability and production. The anti-CD19 and anti-CD3 scFv were directly adopted from the BiTE molecule blinatumomab. The antiCD3 scFv was chosen in the VH-VL orientation, consistent with what used in BiTE. The benchmark molecule was an scFv molecule based on BiTE (v891). AlbuCORE_1 (ABH2) CD3/CD19 fusions were created by attaching the antiCD3 warhead to the natural N terminus of fragment 1 and the antiCD19 to the C terminus of fragment 2 (v1092, polypeptide sequences corresponding to SEQ ID NO:264 and 266). The linkers used were identical to the ones used for the multivalent HER2 AlbuCORE experiments: GGGS (SEQ ID NO: 278) at the N terminus of fragment 1 and (GGSG) 4GG (SEQ ID NO: 279) at the C terminus of fragment 2. A second molecule was created where the warheads were reversed (i.e. anti-CD19 warhead at the natural N terminus of fragment 1 and the anti-CD3 at the C terminus of fragment 2, v1093. v1094 was designed to accommodate two different fusions at the natural termini of the albumin polypeptide (polypeptide sequences corresponding to SEQ ID NO:268). The scFv fusions were linked to the albumin molecule through a GGS linker at the N terminus and a GGSG (SEQ ID NO: 280) linker at the C terminus. The length of the linkers reflect the ones used in the MM-111 molecule, despite having a different sequence type.
Example 11: Heteromultimers with an Albumin Scaffold Bind Specifically to CD3- or CD19-Expressing Cells
[0421] The ability of Anti-CD3×CD19 loaded AlbuCORE-1 (v1092) to CD3+ and CD19+ cells was assessed using FACS and compared to WT-HSA loaded with the same anti-CD scFvs (v1094).
[0422] The results are shown in FIG. 4 and demonstrate that both v1092 and v1094 are able to bind to CD3-expressing Jurkat T-cells and to CD19-expressing Raji B-cells.
Example 12: Heteromultimers with Heterodimeric Fc or Albumin Scaffolds Show Comparable B-Cell Targeting and T-Cell Bridging
Example 13: Exemplary Heteromultimers have Higher Anti-CD3 K.SUB.D .and Higher Bmax in Binding to T and B Cell as Determined by FACS
Example 14: Heteromultimer v875 is Able to Bridge Jurkat CD3 T Cells and Raji CD19 B Cells
Example 15: Bridging of B and T Cells by Heteromultimers is Robust at Varying Antibody Concentrations or Cell Ratios
Example 16: Bridging of B and T Cells is Robust Across Differently Engineered Heteromultimers Constructs
Example 17: Effects of v875, v1380, v1379 on IL-2 Activated and Resting CD20+, CD4+, CD8+Subsets
Example 18: Exemplary Heteromultimers v875 and v873 Require the Presence of Both Effector T Cell and Target B Cells to Mediate Cytotoxic Effects
Example 19: Exemplary Heteromultimers can Mediate ADCC or Impaired ADCC to Target Daudi B Cells
[0444] Antibody-dependent cell-mediated cytotoxicity assays (ADCC) were performed with v875, v1379 and v1380 using Daudi cells as target B cells and FcRy3a immobilized NK92 cells as the effector cells (GS193761) by the following method.
Example 20: Exemplary Heteromultimers have Impaired CDC-Mediated Lysis of Daudi B Cells
[0449] Cell based complement dependent cytotoxicity assays (CDC) were performed with v875, v1379 and v1380 using Daudi cells as target B cells. Human serum from healthy donors (NHS) was used as the source of complement. 10 μl NHS (10% final concentration in 40 reaction volume) were added to each well to initiate the CDC cascade and incubated for 2 hours. Cell viability was measured with CellTiter-Glo@ Luminescent Cell Viability Assay Kit.
Example 21: Cell Proliferation and Cytokine Release Assessment of Exemplary Heteromultimers
Example 22: Exemplary Heteromultimers can Bridge Two or More Target B Cells Per Effector T Cell
[0465] The ability and ratio of numbers of T cell bridged to B cells was examined with the exemplary heteromultimer v875 by microscopy using the method described in Example 3 with the following modifications.
[0466] Labeled Raji B cells (red) and labeled Jurkat T cells (blue) were incubated for 30 min at RT with 3 nM of human IgG or v875. The cell suspension was concentrated by removing 180 μl of supernatant. Cell were resuspended in the remaining volume and imaged at 200× and 400×.
[0467] FIG. 30 shows the results from the T:B cell bridging microscopy comparing v875 and human IgG (3 nM) at 200× and 400× magnification; the phase image (top panel), fluorescence image (middle panel) and inverted fluorescence (bottom panel) are presented. FIG. 30A shows a direct comparison of human IgG and v875 at 200× magnification and illustrates a higher amount of bridging visible between Raji B cell and Jurkat T cells compared to human IgG. FIG. 30B and FIG. 30C show two fields of view for v875 (FIG. 30B) and human IgG (FIG. 30C) at 400× magnification. FIG. 30B shows images of v875-mediated immune complex formation between Jurkat T cells (dark grey cells in fluorescence inverted image) and Raji B cells (light grey cells in fluorescence inverted image), and that one Jurkat T cell can bridge 1-3 Raji B cells. FIG. 30C shows images following human IgG incubation with Jurkat T cells and Raji B cells. FIG. 30C shows an absence of the Jurkat-Raji bridging following incubation with human IgG, compared to the v875-mediated Jurkat: Raji bridging that is visible in FIG. 30B.
Example 23: Exemplary Heteromultimer Binding to Fcγ Receptors as Assessed by Surface Plasmon Resonance
Example 25: Expression and Purification of Heteromultimers
[0476] Description of the methods used in the expression and purification of exemplary heteromultimer are described in Example 2.
Example 26: Exemplary Heteromultimers can be Purified to >99% Heterodimer Purity and <1% Aggregates
Example 27: Exemplary Heteromultimers have a CH3 Tm that is Greater than 75° C.
[0487] CH3 domain stability of exemplary heteromultimers was examined by DSC using the following method. All DSC experiments were carried out using a GE VP-Capillary instrument. The proteins were buffer-exchanged into PBS (pH 7.4) and diluted to 0.3 to 0.7 mg/mL with 0.137 mL loaded into the sample cell and measured with a scan rate of 1° C./min from 20 to 100° C. Data was analyzed using the Origin software (GE Healthcare) with the PBS buffer background subtracted.
[0488] The DSC results shown in FIGS. 33A, B and C show that v875 has an estimated CH3 Tm >76° C. (FIG. 33A), v1380 has an estimated CH3 Tm >82.3° C. (FIG. 33B), and v1379 has an estimated CH3 Tm >82.5° C. (FIG. 33C).
Example 28: Design, Expression and Purification of CD3/CD20 and Additional CD3/CD19 Heteromultimer Constructs
[0489] V5850 (corresponding to polypeptide sequences SEQ ID NOs: 203, 205 and 207), v5851 (corresponding to polypeptide sequences SEQ ID NOs: 209, 211 and 213), v5852 (corresponding to polypeptide sequences SEQ ID NOs: 215, 217 and 219), v6324 (corresponding to polypeptide sequences SEQ ID NOs: 221 and 223), v6325 (corresponding to polypeptide sequences SEQ ID NOs: 225, 227 and 229), v1813 (corresponding to polypeptide sequences SEQ ID NOs: 231, 233 and 235), v1821 (corresponding to polypeptide sequences SEQ ID NOs: 237, 239 and 241), v1823 (corresponding to polypeptide sequences SEQ ID NOs: 243, 245 and 247) exemplify bispecific CD3/CD19 or CD3/CD20 hybrid heterodimeric Fc constructs. Bispecific hybrid variants are composed of a F(ab′) on either chain A or B paired with an scFv-Fc on the alternate polypeptide chain. Chain A of the heterodimer Fc is comprised of the following mutations: T350V_L351Y_F405A_Y407V and Chain B of the heterodimer Fc is comprised of the following mutations: T350V_T366L_K392L_T394W. v6324 exemplifies bispecific CD3/CD20 scFv heterodimeric Fc constructs. V1813, v1821, and v1823 exemplify CD3/CD20 common light chain heterodimeric Fc constructs. Common light chain variants are composed of two different F(ab′) s, each on complimentary heterodimer Fc, which share a single light chain. The specific variant composition is indicated in Table 7.
However, none of the antibodies described in the specification as filed comprises a Fab domain and monospecific, see FIG. 1A, title of the invention.
Regarding antigen-binding domain that binds a different antigen (claim 19), the specification defines antigen as follow:
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The specification discloses just scFv that binds to human CD3 from BITE and scFv that binds to CD19 or CD20 on B cell.
However, the specification does not describe the structure, e.g., amino acid sequence of the heavy and light chain variable region of antigen-binding domain that bind to any CD3 and any and all potential antigen (claim 19) to enable one of skilled in the art to visualize or recognize the member of the genus of the actual claimed dimer and antibody themselves.
The specification does not describe a representative number of species of antibody encompassed by the claims. The specification does not describe the structure correlated with binding falling within the scope of the genus or structural common to the members of the genus so the one of skill in the art can visualize or recognize the member of the genus. An adequate written description must contain enough information about the actual makeup of the claimed products – “a precise definition, such as structure, formula, chemic name, physical properties of other properties, of species falling with the genus sufficient to distinguish the gene from other materials”, which may be present in “functional terminology when the art has established a correlation between structure and function” (Amgen page 1361).
Notably, the specification, does not describe the structure, e.g., amino acid sequence of the heavy and light chain variable domains or the six CDRs that correlated with binding to any antigen (claim 20) and any CD3 complex (claim 21). There is no limitation on the structure and the antigen to which it binds.
At the time of the invention was made, it is known in the art that antibodies have a large repertoire of distinct structures and that a huge variety of antibodies can be made to bind to a single epitope.
For example, Lloyd et al. taught that hundreds of functional antibody fragments can be isolated from an antibody library that bind to the same antigen wherein these antibodies have distinct heavy and light chain sequences (of record, Lloyd et al. Protein Engineering, Design & Selection 22:159-168, 2009, PTO 1449; see, e.g., Discussion).
Similarly, Edwards et al., (of record, J Mol Biol. 334(1): 103-118, 2003; PTO 1449), found that over 1000 antibodies, all different in amino acid sequence, were generated to a single protein; 568 different amino acid sequences identified for the V(H) CDR3 domains of these antibodies (Abstract).
Poosarla et al (of record, Biotechn. Bioeng., 114(6): 1331-1342, 2017; PTO 892) teach substantial diversity in designed mAbs (sharing less than 75% sequence similarity to all existing natural antibody sequences) that bind to the same 12-mer peptide, binding to different epitopes on the same peptide. Said reference further teaches “most B-cell epitopes... in nature consist of residues from different regions of the sequence and are discontinuous...de novo antibody designs against discontinuous epitopes present additional challenge.
Even assuming the antibody binds to CD3, Shepard et al (of record, PLOSOne 18(6): e0273884, June 22, 2023; PTO 892) teach the unpredictability of swapping CD3 binding domains in the context of a BiTE in so far as the binding requirement to CD3 epsilon much less the extracellular epitope:
“We next sought to test our platform for flexibility with respect to screening novel T-cell engaging domains of the bispecific molecule. We developed a number of novel mouse monoclonal antibodies (mAbs) against human CD3 complex using a multi-antigen immunization strategy in mice and traditional hybridoma screening. Through this process, we were able to identify several monoclonal antibodies with reactivity to Jurkat cells (Fig 4A) and human T cells (Fig 4B). To assess whether scFvs derived from novel CD3-targeted mAbs would be functional within as part of a BiTE molecule, we cloned 4 anti-human CD3 single chain variable fragments into CD19 or EGFRvIII specific BiTE plasmids (Fig 4C). We then generated supernatants using transient transfection of HEK293T as described above. BiTE supernatants were screened for activity using Jurkat cells in co-culture with EGFRvIII-expressing U87-VIII targets or CD19-expressing Raji cells. Previously tested constructs using OKT3 CD3-engager arms showed activity with both EGFRvIII and CD19 specific BiTEs, whereas we detected activity for only one of our novel CD3-engager BiTEs and only when combined with an EGFRvIII-specific scFv (Fig 4D). To confirm these results, we repeated BiTE production and Jurkat co-culture screening of CD19 and EGFRvIII targeted BiTE molecules incorporating OKT3 or the novel 1E2 CD3-targeting single chain variable fragment. Whereas EGFRvIII BiTEs incorporating an scFv derived from the 1E2 mAb or OKT3 showed specific reactivity against EGFRvIII expressing U87-vIII cells, only CD19-OKT3 showed reactivity to CD19-expressing Raji cells (Fig 4E). These results indicate that the novel CD3-targeting 1E2-scFv is functional only for an EGFRvIII-targeting BiTE but not a CD19-targeting BiTE, likely due to the specific binding characteristics of the CD19 or EGFRvIII scFv elements.”
“Similarly, we also find that only one of the novel CD3-targeting scFvs tested here showed activity in BiTE format, and even then, activity was restricted to combination with EGFRvIII-targeting scFv and not with a CD19-specific scFv. Again, we have no insight into the failure rate for BiTE molecules in this assay, as the intent of this assay is to provide a rapid means for testing biological activity rather than focusing on various aspects of antibody characterization. We are currently undertaking molecular studies to test whether incorporating different linker domains may be able to improve the activity of BiTEs using the novel CD3-scFv reported to have activity here, something that may provide further insight into the design constraints for these BiTE molecules.”
Given that hundreds of unique antibody structures may bind a single antigen, the structure of an antibody cannot be predicted from the structure of the antigen (as held in Amgen), and a single species, or small group of species, cannot define a structure-function relationship so as to be representative of all the antibodies that bind to that antigen (as held in Abbvie).
Although one of skill in the art could make a mAb against an antigen/CD3 complex by screening a human antibody phage display library, test candidates, and produce a corresponding antibody, note that: “Possession may not be shown by merely describing how to obtain possession of members of the claimed genus or how to identify their common structural features.” See University of Rochester, 358 F.3d at 927, 69 USPQ2d at 1895.
Regarding a first polypeptide construct and a second polypeptide construct each comprises a human IgG1 Fc comprising a variant CH2 domain, each variant CH2 domain comprising a L234A, a L235A, and a D265S (claim 1), the specification discloses a first polypeptide construct and a second polypeptide construct each comprises a human IgG1 Fc wherein the first polypeptide construct (chain A) comprising a L234A, a L235A, a D265S, T350V, L351Y, F405A and Y407V, and wherein the second polypeptide construct (chain B) comprising a L234A, a L235A, a D265S, T350V, T366L, K392L, and T394W, wherein the amino acid positions are numbered according to the EU index of Kabat.
However, the specification does not describe Fc region comprising a first polypeptide construct and a second polypeptide construct each comprises a human IgG1 Fc comprising a variant CH2 domain, each variant CH2 domain comprising just L234A, a L235A, and a D265S capable of forming heterodimer (claim 1).
Vas-Cath Inc. v. Mahurkar, 19 USPQ2d 1111, makes clear that “applicant must convey with reasonable clarity to those skilled in the art that, as of the filing date sought, he or she was in possession of the invention. The invention is, for purposes of the written description inquiry, whatever is now claimed.” (See page 1117.) The specification does not “clearly allow persons of ordinary skill in the art to recognize that [he or she] invented what is claimed.” (See Vas-Cath at page 1116.).
Adequate written description requires more than a mere statement that it is part of the invention and reference to a potential method for isolating it. See Fiers v. Revel, 25 USPQ2d 1601, 1606 (CAFC 1993) and Amgen Inc. v. Chugai Pharmaceutical Co. Ltd., 18 USPQ2d 1016.
One cannot describe what one has not conceived. See Fiddles v. Baird, 30 USPQ2d 1481, 1483. In Fiddles v. Baird, claims directed to mammalian FGF’s were found unpatentable due to lack of written description for the broad class. The specification provided only the bovine sequence. Thus, the specification fails to describe these DNA sequences.
For genus claims, an adequate written description of a claimed genus requires more than a generic statement of an invention's boundaries. A patent must set forth either a representative number of species falling within the scope of the genus or structural features common to the members of the genus. Kubin, Exparte, 83 USPQ2d 1410 (Bd. Pat. App. & Int. 2007); Ariad Pharms., Inc. v. Eli Lilly& Co., 598 F.3d 1336, 1350 (Fed. Cir. 2010).
Therefore, only a bispecific asymmetric heterodimer comprising a first polypeptide construct and a second polypeptide construct, each polypeptide construct comprising a scFv binding domain and a human IgG1 Fc sequence, wherein the first polypeptide construct Fc region sequence and the second polypeptide construct Fc region sequence form a heterodimeric Fc region, wherein the first polypeptide construct comprising a L234A, a L235A, a D265S, T350V, L351Y, F405A and Y407V, and wherein the second polypeptide construct comprising a L234A, a L235A, a D265S, T350V, T366L, K392L, and T394W, wherein the amino acid positions are numbered according to the EU index of Kabat, wherein the scFv binding domain binds to CD3 and the other scFv binding domain binds to CD19 or CD20 on B cell, the antibody is a monoclonal antibody, or a humanized antibody, but not the full breadth of the claims meets the written description provision of 35 U.S.C. § 112, first paragraph.
Applicant is reminded that Vas-Cath makes clear that the written description provision of 35 U.S.C. § 112 is severable from its enablement provision (see page 1115).
Applicant's arguments filed January 14, 2026 have been fully considered but they are not persuasive.
Applicant’s position is that claim 1 has been amended to recite:
An antibody comprising a first polypeptide construct and a second polypeptide construct, each polypeptide construct comprising a human IgG Fc region sequence comprising a variant CH2 domain, each variant CH2 domain comprising a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution, wherein the first polypeptide construct Fc region sequence and the second polypeptide construct Fc region sequence form a heterodimeric Fc region, wherein one of the first and second polypeptide constructs further comprises a CD3 antigen-binding domain that binds to CD3, wherein the CD3 antigen-binding domain is a Fab or an scFv, wherein the IgG is an IgG1, and wherein the amino acid positions are numbered according to the EU index of Kabat.
As stated at MPEP § 2163, "The description need only describe in detail that which is new or not conventional," and the written description requirement is met "[i]f a skilled artisan would have understood the inventor to be in possession of the claimed invention at the time of filing, even if every nuance of the claims is not explicitly described in the specification." Applicant submits that amended claim 1 complies with the written description requirement at least because the claimed elements are described in the application as filed.
First, with respect to the Examiner's assertion that "the disclosure of just heterodimer is not representative of all dimers, e.g., homodimer or heterodimer....," claim 1 has been amended to recite that "the first polypeptide construct Fc region sequence and the second polypeptide construct Fc region sequence form a heterodimeric Fc region." Support for this amendment can be found throughout the application as filed, e.g., at paragraphs [0007]-[0008] and [0089]. Thus, the amended claims are directed to heterodimers, as described in the application as filed.
Second, regarding the Examiner's assertion that "the specification does not describe the structure, e.g., amino acid sequence of the heavy and light chain variable region of all antigen- binding domain... to enable one of skilled in the art to visualize or recognize the member of the genus of the actual claimed dimer and antibody themselves," Applicant respectfully disagrees.
The invention as claimed in amended claim 1 relates to a specific combination of mutations (a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution) that are incorporated into the CH2 domain of an antibody IgG Fc region. These mutations can silence FcyR binding. Amended claim 1 further specifies that the IgG Fc region is an IgG1 Fc region and that the antibody comprises an antigen-binding domain that binds to CD3.
At the time the present application was filed, the skilled person would understand that FcyR binding is mediated by the Fc region of an antibody and that the effect of silencing FcyR binding by the mutations recited in amended claim 1 is, therefore, independent of the heavy and light chain variable region sequences of the antigen-binding domains. As such, the skilled person would appreciate that various antigen-binding domain sequences known in the art could be included in the claimed antibody and the antibody would retain abrogated FcyR binding.
At the time the present application was filed, the sequence of the IgG1 Fc region was well-known in the art and the unique combination of CH2 domain mutations that abrogate FcyR binding are specifically defined in amended claim 1 and in the application as filed.
Additionally, the sequences of antigen-binding domains of various antibodies, including anti-CD3 antibodies, were also publicly available at the time the instant application was filed and thus formed part of the state of the art. For example, the Therapeutic Antibody Database (Tabs), established by Craic Computing LLC on June 1, 2011, provided protein sequences for more than 950 antibodies as of its launch date (see webpage a copy of which is provided herewith). Given that these structures were neither new nor unconventional, Applicant submits that it would not have been necessary for the present application to describe all the structures of the instantly claimed antigen-binding domains, and it was sufficient for the present claims to describe the antigen-binding domains as instantly claimed.
In summary, Applicant submits that the skilled artisan having regard to the knowledge available in the art at the time of filing would recognize that the inventors were in possession of the claimed invention at the time of filing. Thus, claim 1 and claims dependent therefrom meet the requirements for written description. Accordingly, reconsideration and withdrawal of the rejection is respectfully requested.
In response, the amendment to claim 1 is acknowledged.
The claim encompass an antibody comprising a first polypeptide construct and a second polypeptide construct, each polypeptide construct comprising a human IgG Fc region sequence comprising a variant CH2 domain, each variant CH2 domain comprising a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution,
wherein the first polypeptide construct Fc region sequence and the second polypeptide construct Fc region sequence form a heterodimeric Fc region,
wherein one of the first and second polypeptide constructs further comprises a CD3 antigen-binding domain that binds to CD3, wherein the CD3 antigen-binding domain is a Fab or an scFv,
wherein the human IgG Fc region is an IgG1 Fc region, and
wherein the amino acid positions are numbered according to the EU index of Kabat.
The specification discloses bispecific asymmetric heterodimer comprising having the following structure:
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It should be noted that the antigen-binding domain that binds to CD3 is from just anti-CD3 scFv called BITE.
However, the specification does not describe antibody comprising a first polypeptide construct and a second polypeptide construct, each polypeptide construct comprising a human IgG Fc region sequence comprising a variant CH2 domain, each variant CH2 domain comprising just a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution,
wherein the first polypeptide construct Fc region sequence and the second polypeptide construct Fc region sequence form a heterodimeric Fc region,
wherein one of the first and second polypeptide constructs further comprises a CD3 antigen-binding domain that binds to CD3, wherein the CD3 antigen-binding domain is a Fab,
wherein the human IgG Fc region is an IgG1 Fc region, and
wherein the amino acid positions are numbered according to the EU index of Kabat, this is new matter.
However, none of the antibodies described in the specification as filed comprises a Fab domain and monospecific, see FIG. 1A, title of the invention.
Regarding antigen-binding domain that binds a different antigen (claim 19), the specification defines antigen as follow:
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The specification discloses just scFv that binds to human CD3 from BITE and scFv that binds to CD19 or CD20 on B cell, see para. [0359] to [0366], [0489] above.
However, the specification does not describe the structure, e.g., amino acid sequence of the heavy and light chain variable region of antigen-binding domain that bind to any CD3 and any and all potential antigen (claim 19) to enable one of skilled in the art to visualize or recognize the member of the genus of the actual claimed dimer and antibody themselves.
The specification does not describe a representative number of species of antibody encompassed by the claims. The specification does not describe the structure correlated with binding falling within the scope of the genus or structural common to the members of the genus so the one of skill in the art can visualize or recognize the member of the genus. An adequate written description must contain enough information about the actual makeup of the claimed products – “a precise definition, such as structure, formula, chemic name, physical properties of other properties, of species falling with the genus sufficient to distinguish the gene from other materials”, which may be present in “functional terminology when the art has established a correlation between structure and function” (Amgen page 1361).
Notably, the specification, does not describe the structure, e.g., amino acid sequence of the heavy and light chain variable domains or the six CDRs that correlated with binding to any antigen (claim 20) and any CD3 complex (claim 21). There is no limitation on the structure and the antigen to which it binds.
At the time of the invention was made, it is known in the art that antibodies have a large repertoire of distinct structures and that a huge variety of antibodies can be made to bind to a single epitope.
For example, Lloyd et al. taught that hundreds of functional antibody fragments can be isolated from an antibody library that bind to the same antigen wherein these antibodies have distinct heavy and light chain sequences (of record, Lloyd et al. Protein Engineering, Design & Selection 22:159-168, 2009, PTO 1449; see, e.g., Discussion).
Similarly, Edwards et al., (of record, J Mol Biol. 334(1): 103-118, 2003; PTO 1449), found that over 1000 antibodies, all different in amino acid sequence, were generated to a single protein; 568 different amino acid sequences identified for the V(H) CDR3 domains of these antibodies (Abstract).
Poosarla et al (of record, Biotechn. Bioeng., 114(6): 1331-1342, 2017; PTO 892) teach substantial diversity in designed mAbs (sharing less than 75% sequence similarity to all existing natural antibody sequences) that bind to the same 12-mer peptide, binding to different epitopes on the same peptide. Said reference further teaches “most B-cell epitopes... in nature consist of residues from different regions of the sequence and are discontinuous...de novo antibody designs against discontinuous epitopes present additional challenge.
Even assuming the antibody binds to CD3, Shepard et al (of record, PLOSOne 18(6): e0273884, June 22, 2023; PTO 892) teach the unpredictability of swapping CD3 binding domains in the context of a BiTE in so far as the binding requirement to CD3 epsilon much less the extracellular epitope:
“We next sought to test our platform for flexibility with respect to screening novel T-cell engaging domains of the bispecific molecule. We developed a number of novel mouse monoclonal antibodies (mAbs) against human CD3 complex using a multi-antigen immunization strategy in mice and traditional hybridoma screening. Through this process, we were able to identify several monoclonal antibodies with reactivity to Jurkat cells (Fig 4A) and human T cells (Fig 4B). To assess whether scFvs derived from novel CD3-targeted mAbs would be functional within as part of a BiTE molecule, we cloned 4 anti-human CD3 single chain variable fragments into CD19 or EGFRvIII specific BiTE plasmids (Fig 4C). We then generated supernatants using transient transfection of HEK293T as described above. BiTE supernatants were screened for activity using Jurkat cells in co-culture with EGFRvIII-expressing U87-VIII targets or CD19-expressing Raji cells. Previously tested constructs using OKT3 CD3-engager arms showed activity with both EGFRvIII and CD19 specific BiTEs, whereas we detected activity for only one of our novel CD3-engager BiTEs and only when combined with an EGFRvIII-specific scFv (Fig 4D). To confirm these results, we repeated BiTE production and Jurkat co-culture screening of CD19 and EGFRvIII targeted BiTE molecules incorporating OKT3 or the novel 1E2 CD3-targeting single chain variable fragment. Whereas EGFRvIII BiTEs incorporating an scFv derived from the 1E2 mAb or OKT3 showed specific reactivity against EGFRvIII expressing U87-vIII cells, only CD19-OKT3 showed reactivity to CD19-expressing Raji cells (Fig 4E). These results indicate that the novel CD3-targeting 1E2-scFv is functional only for an EGFRvIII-targeting BiTE but not a CD19-targeting BiTE, likely due to the specific binding characteristics of the CD19 or EGFRvIII scFv elements.”
“Similarly, we also find that only one of the novel CD3-targeting scFvs tested here showed activity in BiTE format, and even then, activity was restricted to combination with EGFRvIII-targeting scFv and not with a CD19-specific scFv. Again, we have no insight into the failure rate for BiTE molecules in this assay, as the intent of this assay is to provide a rapid means for testing biological activity rather than focusing on various aspects of antibody characterization. We are currently undertaking molecular studies to test whether incorporating different linker domains may be able to improve the activity of BiTEs using the novel CD3-scFv reported to have activity here, something that may provide further insight into the design constraints for these BiTE molecules.”
Given that hundreds of unique antibody structures may bind a single antigen, the structure of an antibody cannot be predicted from the structure of the antigen (as held in Amgen), and a single species, or small group of species, cannot define a structure-function relationship so as to be representative of all the antibodies that bind to that antigen (as held in Abbvie).
Although one of skill in the art could make a mAb against an antigen/CD3 complex by screening a human antibody phage display library, test candidates, and produce a corresponding antibody, note that: “Possession may not be shown by merely describing how to obtain possession of members of the claimed genus or how to identify their common structural features.” See University of Rochester, 358 F.3d at 927, 69 USPQ2d at 1895.
Regarding a first polypeptide construct and a second polypeptide construct each comprises a human IgG1 Fc comprising a variant CH2 domain, each variant CH2 domain comprising a L234A, a L235A, and a D265S (claim 1), the specification discloses a first polypeptide construct and a second polypeptide construct each comprises a human IgG1 Fc wherein the first polypeptide construct (chain A) comprising a L234A, a L235A, a D265S, T350V, L351Y, F405A and Y407V, and wherein the second polypeptide construct (chain B) comprising a L234A, a L235A, a D265S, T350V, T366L, K392L, and T394W, wherein the amino acid positions are numbered according to the EU index of Kabat, see para. [0359] to [0366], [0489] above.
However, the specification does not describe heterodimeric Fc region comprising a first polypeptide construct and a second polypeptide construct each comprises a human IgG1 Fc comprising a variant CH2 domain, each variant CH2 domain comprising just L234A, a L235A, and a D265S (claim 1) capable of forming heterodimer. Substitutions of L234A, a L235A, and a D265S in the CH2 domain do not form Fc heterodimer. It is the combination of substitution in the CH3 domain such as the first polypeptide construct (chain A) comprising a T350V, L351Y, F405A and Y407V, and wherein the second polypeptide construct (chain B) comprising a T350V, T366L, K392L, and T394W that forms stable heterodimer.
For these reasons, the rejection is maintained.
Claims 1, 3-4, 6, 9-11, 14 and 19 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for a bispecific asymmetric heterodimer comprising anti-CD19 and CD3 scFvs fused to an IgG1 heterodimer comprising a first Fc polypeptide and a second Fc polypeptide, each Fc polypeptide comprising a human CH2 domain, each CH2 domain comprises a L234A amino acid substitution, a L235A amino acid substitution and a D265S amino acid substitution, wherein the amino acid positions are numbered according to the EU index of Kabat, wherein the first and second polypeptides comprise the amino acid sequences selected from the group consisting of SEQ ID NO: 38 and SEQ ID NO: 40, SEQ ID NO: 42 and SEQ ID NO: 44, SEQ ID NO: 46 and SEQ ID NO: 48, SEQ ID NO: 66 and SEQ NO: 68, SEQ ID NO: 70 and SEQ NO: 72, SEQ ID NO: 74 and SEQ NO: 76, SEQ ID NO: 78 and SEQ NO: 80, SEQ ID NO: 82 and SEQ NO: 84, SEQ ID NO: 133 and SEQ ID NO: 134, SEQ ID NO: 137 and SEQ ID NO: 138, and SEQ ID NO: 141 and SEQ ID NO: 142, does not reasonably provide enablement for the isolated dimer set forth in claims 1-14 and 19-21. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make/use the invention commensurate in scope with these claims.
Claim 1 encompasses an antibody comprising a first polypeptide construct and a second polypeptide construct, each polypeptide construct comprising a human IgG Fc region sequence comprising a variant CH2 domain, each variant CH2 domain comprising a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution,
wherein the first polypeptide construct Fc region sequence and the second polypeptide construct Fc region sequence form a heterodimeric Fc region,
wherein one of the first and second polypeptide constructs further comprises a CD3 antigen-binding domain that binds to CD3, wherein the CD3 antigen-binding domain is a Fab or an scFv,
wherein the human IgG Fc region is an IgG1 Fc region, and
wherein the amino acid positions are numbered according to the EU index of Kabat.
Claim 3 encompasses the isolated dimer of claim 2, wherein the antigen-binding domain comprises an scFv.
Claim 4 encompasses the isolated dimer of claim 2, wherein the antigen-binding domain comprises a Fab.
Claim 6 encompasses the isolated dimer of claim 1, wherein one of the first and second polypeptide further comprises amino acid substitutions T350V, L351Y, F405A, and Y407V, and the other of the polypeptides further comprises amino acid substitutions T350V, T366L, K392L, and T394W.
Claim 9 encompasses the e antibody of claim 1, wherein the antibody is any monoclonal antibody, or any humanized antibody.
Claim 10 encompasses the antibody of claim 1, wherein the antibody is multispecific.
Claim 11 encompasses the antibody of claim 1, wherein the antibody is bispecific.
Claim 14 encompasses a pharmaceutical composition comprising the antibody of claim 1 and a pharmaceutically acceptable carrier.
Claim 19 encompasses the isolated dimer of claim 1, wherein each of the first and second polypeptide construct further comprises an antigen-binding domain, and wherein each antigen-binding domain binds a different antigen.
The specification discloses bispecific asymmetric heterodimer comprising having the following structure:
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It should be noted that the antigen-binding domain that binds to CD3 is from just anti-CD3 scFv called BITE.
Regarding antigen-binding domain that binds a different antigen (claim 19), the specification defines antigen as follow:
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The specification discloses just scFv that binds to human CD3 from BITE and scFv that binds to CD19 or CD20 on B cell and reproduced below.
[0359] v4541 has an anti-CD3 BiTE™ (VH-VL) 44-100SS scFv on chain A and anti-CD19 BiTE™ (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 50 and 52] Cyno/Human Cross-Reactive Anti-CD3 and Anti-CD19 Bispecific Fc Knock-Out Variants with or without Disulfide 44-100 Stabilization
[0360] v4542 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VH-VL) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 54 and 56].
[0361] v4543 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VH-VL) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 58 and 60]
[0362] v4544 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 62 and 64]
[0363] v4545 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VL-VH) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 66 and 68]
[0364] v4546 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VH-VL) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 70 and 72]
[0365] v4547 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VH-VL) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 74 and 76].
[0366] v4548 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 78 and 80] v4549 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VL-VH) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 82 and 84]
Example 3: Heteromultimer v873 is Able to Bridge Jurkat CD3 T Cells and Raji CD19 B Cells
Example 4: Heteromultimers Bind Selectively to CD3- and CD19-Expressing Cells
[0384] The ability of an exemplary heteromultimer, v873, to bind specifically to CD3 and CD19 was assessed by FACS. One-armed antibodies (OAAs) against CD3 and CD19 were also prepared as described in Example 2 and tested as controls in the whole cell FACS binding assay described below.
Example 5: Heteromultimers Mediate PBMC Killing of Target Raji B Cells
Example 6: Heteromultimers Mediate Redirected Killing of Target Raji B Cells with Resting and IL-2 Activated CD4+ and CD8+ T Cells
[0405] The ability of an exemplary heteromultimers, v875, v1379, v1380 to mediate CD4+ and CD8+ T cell cytoxicity against target Raji B-cells was measured as described in Example 5.
Example 7: The Heterodimeric Fc Contributes to Target Raji B Cell Cytotoxicity
[0410] The ability of exemplary heteromultimers, v875, and v873, to mediate target Raji B-cell cytotoxicity in the presence and absence of Fc was measured as described in Example 5.
Example 8: Heteromultimers Mediate Autologous B Cell Cytotoxicity
[0413] The ability of exemplary the heteromultimer v875 and v873 to kill autologous B-cells was measured in total and resting IL-2-stimulated PBMCs where the percent of CD19+ 7AAD+ cells following incubation with v875 and v873 (300 nM, n=3 donors) was determined by flow cytometry as described in Example 5.
Example 9: Heteromultimer v875 Spares Autologous T Cell Cytotoxicity Compared to BiTE
[0415] The effects of exemplary heteromultimer v875 and v873 treatment on the autologous T cell population was assessed in total and resting IL-2-stimulated PBMCs where the percent of CD3+ 7AAD+ cells following incubation with v875 and v873 (300 nM, n=3 donors) was determined by flow cytometry as described in Example 5.
[0416] FIG. 22 shows, relative to untreated media and human IgG controls, v875 has a more selective B cell killing by sparing more autologous T cells compared to v873 and v891.
Example 10: Design, Expression and Purification of Heteromultimers with an Albumin Scaffold
[0417] The following exemplary CD3-CD19 binding heteromultimers based on an albumin scaffold were designed and prepared as follows.
[0418] The sequences for the anti-CD19 and anti-CD3 scFvs were chosen from two molecules that are currently in clinical trials and are well documented and tested for stability and production. The anti-CD19 and anti-CD3 scFv were directly adopted from the BiTE molecule blinatumomab. The antiCD3 scFv was chosen in the VH-VL orientation, consistent with what used in BiTE. The benchmark molecule was an scFv molecule based on BiTE (v891). AlbuCORE_1 (ABH2) CD3/CD19 fusions were created by attaching the antiCD3 warhead to the natural N terminus of fragment 1 and the antiCD19 to the C terminus of fragment 2 (v1092, polypeptide sequences corresponding to SEQ ID NO:264 and 266). The linkers used were identical to the ones used for the multivalent HER2 AlbuCORE experiments: GGGS (SEQ ID NO: 278) at the N terminus of fragment 1 and (GGSG) 4GG (SEQ ID NO: 279) at the C terminus of fragment 2. A second molecule was created where the warheads were reversed (i.e. anti-CD19 warhead at the natural N terminus of fragment 1 and the anti-CD3 at the C terminus of fragment 2, v1093. v1094 was designed to accommodate two different fusions at the natural termini of the albumin polypeptide (polypeptide sequences corresponding to SEQ ID NO:268). The scFv fusions were linked to the albumin molecule through a GGS linker at the N terminus and a GGSG (SEQ ID NO: 280) linker at the C terminus. The length of the linkers reflect the ones used in the MM-111 molecule, despite having a different sequence type.
Example 11: Heteromultimers with an Albumin Scaffold Bind Specifically to CD3- or CD19-Expressing Cells
[0421] The ability of Anti-CD3×CD19 loaded AlbuCORE-1 (v1092) to CD3+ and CD19+ cells was assessed using FACS and compared to WT-HSA loaded with the same anti-CD scFvs (v1094).
[0422] The results are shown in FIG. 4 and demonstrate that both v1092 and v1094 are able to bind to CD3-expressing Jurkat T-cells and to CD19-expressing Raji B-cells.
Example 12: Heteromultimers with Heterodimeric Fc or Albumin Scaffolds Show Comparable B-Cell Targeting and T-Cell Bridging
Example 13: Exemplary Heteromultimers have Higher Anti-CD3 K.SUB.D .and Higher Bmax in Binding to T and B Cell as Determined by FACS
Example 14: Heteromultimer v875 is Able to Bridge Jurkat CD3 T Cells and Raji CD19 B Cells
Example 15: Bridging of B and T Cells by Heteromultimers is Robust at Varying Antibody Concentrations or Cell Ratios
Example 16: Bridging of B and T Cells is Robust Across Differently Engineered Heteromultimers Constructs
Example 17: Effects of v875, v1380, v1379 on IL-2 Activated and Resting CD20+, CD4+, CD8+Subsets
Example 18: Exemplary Heteromultimers v875 and v873 Require the Presence of Both Effector T Cell and Target B Cells to Mediate Cytotoxic Effects
Example 19: Exemplary Heteromultimers can Mediate ADCC or Impaired ADCC to Target Daudi B Cells
[0444] Antibody-dependent cell-mediated cytotoxicity assays (ADCC) were performed with v875, v1379 and v1380 using Daudi cells as target B cells and FcRy3a immobilized NK92 cells as the effector cells (GS193761) by the following method.
Example 20: Exemplary Heteromultimers have Impaired CDC-Mediated Lysis of Daudi B Cells
[0449] Cell based complement dependent cytotoxicity assays (CDC) were performed with v875, v1379 and v1380 using Daudi cells as target B cells. Human serum from healthy donors (NHS) was used as the source of complement. 10 μl NHS (10% final concentration in 40 reaction volume) were added to each well to initiate the CDC cascade and incubated for 2 hours. Cell viability was measured with CellTiter-Glo@ Luminescent Cell Viability Assay Kit.
Example 21: Cell Proliferation and Cytokine Release Assessment of Exemplary Heteromultimers
Example 22: Exemplary Heteromultimers can Bridge Two or More Target B Cells Per Effector T Cell
[0465] The ability and ratio of numbers of T cell bridged to B cells was examined with the exemplary heteromultimer v875 by microscopy using the method described in Example 3 with the following modifications.
[0466] Labeled Raji B cells (red) and labeled Jurkat T cells (blue) were incubated for 30 min at RT with 3 nM of human IgG or v875. The cell suspension was concentrated by removing 180 μl of supernatant. Cell were resuspended in the remaining volume and imaged at 200× and 400×.
[0467] FIG. 30 shows the results from the T:B cell bridging microscopy comparing v875 and human IgG (3 nM) at 200× and 400× magnification; the phase image (top panel), fluorescence image (middle panel) and inverted fluorescence (bottom panel) are presented. FIG. 30A shows a direct comparison of human IgG and v875 at 200× magnification and illustrates a higher amount of bridging visible between Raji B cell and Jurkat T cells compared to human IgG. FIG. 30B and FIG. 30C show two fields of view for v875 (FIG. 30B) and human IgG (FIG. 30C) at 400× magnification. FIG. 30B shows images of v875-mediated immune complex formation between Jurkat T cells (dark grey cells in fluorescence inverted image) and Raji B cells (light grey cells in fluorescence inverted image), and that one Jurkat T cell can bridge 1-3 Raji B cells. FIG. 30C shows images following human IgG incubation with Jurkat T cells and Raji B cells. FIG. 30C shows an absence of the Jurkat-Raji bridging following incubation with human IgG, compared to the v875-mediated Jurkat: Raji bridging that is visible in FIG. 30B.
Example 23: Exemplary Heteromultimer Binding to Fcγ Receptors as Assessed by Surface Plasmon Resonance
Example 25: Expression and Purification of Heteromultimers
[0476] Description of the methods used in the expression and purification of exemplary heteromultimer are described in Example 2.
Example 26: Exemplary Heteromultimers can be Purified to >99% Heterodimer Purity and <1% Aggregates
Example 27: Exemplary Heteromultimers have a CH3 Tm that is Greater than 75° C.
[0487] CH3 domain stability of exemplary heteromultimers was examined by DSC using the following method. All DSC experiments were carried out using a GE VP-Capillary instrument. The proteins were buffer-exchanged into PBS (pH 7.4) and diluted to 0.3 to 0.7 mg/mL with 0.137 mL loaded into the sample cell and measured with a scan rate of 1° C./min from 20 to 100° C. Data was analyzed using the Origin software (GE Healthcare) with the PBS buffer background subtracted.
[0488] The DSC results shown in FIGS. 33A, B and C show that v875 has an estimated CH3 Tm >76° C. (FIG. 33A), v1380 has an estimated CH3 Tm >82.3° C. (FIG. 33B), and v1379 has an estimated CH3 Tm >82.5° C. (FIG. 33C).
Example 28: Design, Expression and Purification of CD3/CD20 and Additional CD3/CD19 Heteromultimer Constructs
[0489] V5850 (corresponding to polypeptide sequences SEQ ID NOs: 203, 205 and 207), v5851 (corresponding to polypeptide sequences SEQ ID NOs: 209, 211 and 213), v5852 (corresponding to polypeptide sequences SEQ ID NOs: 215, 217 and 219), v6324 (corresponding to polypeptide sequences SEQ ID NOs: 221 and 223), v6325 (corresponding to polypeptide sequences SEQ ID NOs: 225, 227 and 229), v1813 (corresponding to polypeptide sequences SEQ ID NOs: 231, 233 and 235), v1821 (corresponding to polypeptide sequences SEQ ID NOs: 237, 239 and 241), v1823 (corresponding to polypeptide sequences SEQ ID NOs: 243, 245 and 247) exemplify bispecific CD3/CD19 or CD3/CD20 hybrid heterodimeric Fc constructs. Bispecific hybrid variants are composed of a F(ab′) on either chain A or B paired with an scFv-Fc on the alternate polypeptide chain. Chain A of the heterodimer Fc is comprised of the following mutations: T350V_L351Y_F405A_Y407V and Chain B of the heterodimer Fc is comprised of the following mutations: T350V_T366L_K392L_T394W. v6324 exemplifies bispecific CD3/CD20 scFv heterodimeric Fc constructs. V1813, v1821, and v1823 exemplify CD3/CD20 common light chain heterodimeric Fc constructs. Common light chain variants are composed of two different F(ab′) s, each on complimentary heterodimer Fc, which share a single light chain. The specific variant composition is indicated in Table 7.
However, the specification does not teach the structure, e.g., amino acid sequence of the heavy and light chain variable region of all antigen-binding domain, e.g., any scFv (claim 3), Fab (claim 4), antibody (claim 8), (humanized or monoclonal antibody (claim 9), multispecific antibody (claim 10) or bispecific antibody (claim 11) that bind to any and all potential different antigen (claim 19) and all CD3 as a pharmaceutical composition (claim 14) to enable one of skilled in the art to make and use without undue experimentation.
It is known in the art that antibodies have a large repertoire of distinct structures and that a huge variety of antibodies can be made to bind to a single epitope.
For example, Lloyd et al (of record, Protein Engineering, Design & Selection 22:159-168, 2009; PTO 1449) teach that hundreds of functional antibody fragments can be isolated from an antibody library that bind to the same antigen wherein these antibodies have distinct heavy and light chain sequences, see, e.g., Discussion.
Similarly, Edwards et al (of record, J Mol Biol 334(1): 103-118, 2003; PTO 1449) find that over 1000 antibodies, all different in amino acid sequence, were generated to a single protein; 568 different amino acid sequences identified for the V(H) CDR3 domains of these antibodies (Abstract).
Poosarla et al (of record, Biotechn. Bioeng., 114(6): 1331-1342, 2017; PTO 892) teach substantial diversity in designed mAbs (sharing less than 75% sequence similarity to all existing natural antibody sequences) that bind to the same 12-mer peptide, binding to different epitopes on the same peptide. Said reference further teaches “most B-cell epitopes... in nature consist of residues from different regions of the sequence and are discontinuous...de novo antibody designs against discontinuous epitopes present additional challenge.
Even assuming the antibody binds to CD3, Shepard et al (of record, PLOSOne 18(6): e0273884, June 22, 2023; PTO 892) teach the unpredictability of swapping CD3 binding domains in the context of a BiTE in so far as the binding requirement to CD3 epsilon much less the extracellular epitope:
“We next sought to test our platform for flexibility with respect to screening novel T-cell engaging domains of the bispecific molecule. We developed a number of novel mouse monoclonal antibodies (mAbs) against human CD3 complex using a multi-antigen immunization strategy in mice and traditional hybridoma screening. Through this process, we were able to identify several monoclonal antibodies with reactivity to Jurkat cells (Fig 4A) and human T cells (Fig 4B). To assess whether scFvs derived from novel CD3-targeted mAbs would be functional within as part of a BiTE molecule, we cloned 4 anti-human CD3 single chain variable fragments into CD19 or EGFRvIII specific BiTE plasmids (Fig 4C). We then generated supernatants using transient transfection of HEK293T as described above. BiTE supernatants were screened for activity using Jurkat cells in co-culture with EGFRvIII-expressing U87-VIII targets or CD19-expressing Raji cells. Previously tested constructs using OKT3 CD3-engager arms showed activity with both EGFRvIII and CD19 specific BiTEs, whereas we detected activity for only one of our novel CD3-engager BiTEs and only when combined with an EGFRvIII-specific scFv (Fig 4D). To confirm these results, we repeated BiTE production and Jurkat co-culture screening of CD19 and EGFRvIII targeted BiTE molecules incorporating OKT3 or the novel 1E2 CD3-targeting single chain variable fragment. Whereas EGFRvIII BiTEs incorporating an scFv derived from the 1E2 mAb or OKT3 showed specific reactivity against EGFRvIII expressing U87-vIII cells, only CD19-OKT3 showed reactivity to CD19-expressing Raji cells (Fig 4E). These results indicate that the novel CD3-targeting 1E2-scFv is functional only for an EGFRvIII-targeting BiTE but not a CD19-targeting BiTE, likely due to the specific binding characteristics of the CD19 or EGFRvIII scFv elements.”
“Similarly, we also find that only one of the novel CD3-targeting scFvs tested here showed activity in BiTE format, and even then, activity was restricted to combination with EGFRvIII-targeting scFv and not with a CD19-specific scFv. Again, we have no insight into the failure rate for BiTE molecules in this assay, as the intent of this assay is to provide a rapid means for testing biological activity rather than focusing on various aspects of antibody characterization. We are currently undertaking molecular studies to test whether incorporating different linker domains may be able to improve the activity of BiTEs using the novel CD3-scFv reported to have activity here, something that may provide further insight into the design constraints for these BiTE molecules.”
Given that hundreds of unique antibody structures may bind a single antigen, the structure of an antibody cannot be predicted from the structure of the antigen, and a single species, or small group of species, cannot define a structure-function relationship so as to be representative of all the antibodies that bind to that antigen.
Regarding pharmaceutical composition (claim 14), the specification does not teach a representative number of species of antibody that correlated with binding to which antigen is effective as a pharmaceutical composition for treating which disease. There are insufficient in vivo working example. It is unpredictable which dimer or antibody is effective as a pharmaceutical composition for treating which disease. Thus, the scope of the claims is extremely broad compared to the guidance and exemplification provided in the specification.
The scope of the claims must bear a reasonable correlation with the scope of enablement. See In re Fisher, 166 USPQ 19 24 (CCPA 1970).
In view of the lack of the predictability of the art to which the invention pertains as evidenced by Lloyd et al, Edwards, Poosarla and Shepard et al, the lack of guidance and direction provided by applicant, and the absence of in vivo working examples, it would require undue experimentation of one skilled in the art to practice the claimed invention, commensurate in scope with the claims. See page 1338, footnote 7 of Ex parte Aggarwal, 23 USPQ2d 1334 (PTO Bd. Pat App. & Inter. 1992).
In re wands, 858 F.2d at 737, 8 USPQ2d at 1404 (Fed. Cir. 1988), the decision of the court indicates that the more unpredictable the area is, the more specific enablement is necessary.
Applicant's arguments filed January 14, 2026 have been fully considered but they are not persuasive.
Applicant submits that the claims are enabled at least because the application as filed and amended claim 1 define a general quality running through the class that gives it a peculiar fitness for the particular purpose and that reliably enables a skilled artisan to make and use the claimed invention.
The invention as claimed in amended claim 1 relates to a specific combination of mutations (a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution) that, when incorporated into the CH2 domain of an antibody, silence FcyR binding. Amended claim 1 recites an antibody "comprising a first polypeptide construct and a second polypeptide construct, each polypeptide construct comprising a human IgG Fc region sequence comprising a variant CH2 domain, each variant CH2 domain comprising a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution." As outlined above, FcyR binding is mediated by the Fc region of an antibody and the effect of silencing FcyR binding by the mutations recited in amended claim 1, therefore, is independent of the heavy and light chain variable region sequences of the antigen-binding domains. These mutations are a general quality running through the class that gives it a peculiar fitness for the particular purpose-silencing the FcyR binding.
Additionally, the sequence of the human IgG1 Fc region was well-known in the art at the time the application was filed and the specific mutations that abrogate FcyR binding are defined in claim 1 and the specification as filed both by position and by the nature of each amino acid substitution.
Given that the specification teaches that modification of the CH2 domain in an IgG1 Fc region to include the specific mutations L234A, L235A and D265S imparts the property of abrogated FcyR binding to an antibody, the general quality that imparts the property of abrogated FcyR binding to an antibody, as instantly claimed, is defined structurally in the specification and one skilled in the art could readily introduce these mutations into the Fc region of an antibody in order to silence FcyR binding, thereby reliably making or using the claimed invention.
At least for the foregoing reasons, amended claim 1 and claims dependent therefrom were enabled by the application as filed. Accordingly, reconsideration and withdrawal of the rejection is respectfully requested.
In response, the claims are not enabled because of the following reasons.
First, regarding antigen-binding domain that binds any CD3 (claim 1) and a different antigen (claim 19), the specification defines antigen as follow:
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The specification discloses just scFv that binds to human CD3 from BITE and scFv that binds to CD19 or CD20 on B cell and reproduced below.
[0359] v4541 has an anti-CD3 BiTE™ (VH-VL) 44-100SS scFv on chain A and anti-CD19 BiTE™ (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 50 and 52] Cyno/Human Cross-Reactive Anti-CD3 and Anti-CD19 Bispecific Fc Knock-Out Variants with or without Disulfide 44-100 Stabilization
[0360] v4542 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VH-VL) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 54 and 56].
[0361] v4543 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VH-VL) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 58 and 60]
[0362] v4544 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 62 and 64]
[0363] v4545 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VL-VH) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 66 and 68]
[0364] v4546 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VH-VL) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 70 and 72]
[0365] v4547 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VH-VL) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 74 and 76].
[0366] v4548 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 78 and 80] v4549 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VL-VH) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 82 and 84]
Example 3: Heteromultimer v873 is Able to Bridge Jurkat CD3 T Cells and Raji CD19 B Cells
Example 4: Heteromultimers Bind Selectively to CD3- and CD19-Expressing Cells
[0384] The ability of an exemplary heteromultimer, v873, to bind specifically to CD3 and CD19 was assessed by FACS. One-armed antibodies (OAAs) against CD3 and CD19 were also prepared as described in Example 2 and tested as controls in the whole cell FACS binding assay described below.
Example 5: Heteromultimers Mediate PBMC Killing of Target Raji B Cells
Example 6: Heteromultimers Mediate Redirected Killing of Target Raji B Cells with Resting and IL-2 Activated CD4+ and CD8+ T Cells
[0405] The ability of an exemplary heteromultimers, v875, v1379, v1380 to mediate CD4+ and CD8+ T cell cytoxicity against target Raji B-cells was measured as described in Example 5.
Example 7: The Heterodimeric Fc Contributes to Target Raji B Cell Cytotoxicity
[0410] The ability of exemplary heteromultimers, v875, and v873, to mediate target Raji B-cell cytotoxicity in the presence and absence of Fc was measured as described in Example 5.
Example 8: Heteromultimers Mediate Autologous B Cell Cytotoxicity
[0413] The ability of exemplary the heteromultimer v875 and v873 to kill autologous B-cells was measured in total and resting IL-2-stimulated PBMCs where the percent of CD19+ 7AAD+ cells following incubation with v875 and v873 (300 nM, n=3 donors) was determined by flow cytometry as described in Example 5.
Example 9: Heteromultimer v875 Spares Autologous T Cell Cytotoxicity Compared to BiTE
[0415] The effects of exemplary heteromultimer v875 and v873 treatment on the autologous T cell population was assessed in total and resting IL-2-stimulated PBMCs where the percent of CD3+ 7AAD+ cells following incubation with v875 and v873 (300 nM, n=3 donors) was determined by flow cytometry as described in Example 5.
[0416] FIG. 22 shows, relative to untreated media and human IgG controls, v875 has a more selective B cell killing by sparing more autologous T cells compared to v873 and v891.
Example 10: Design, Expression and Purification of Heteromultimers with an Albumin Scaffold
[0417] The following exemplary CD3-CD19 binding heteromultimers based on an albumin scaffold were designed and prepared as follows.
[0418] The sequences for the anti-CD19 and anti-CD3 scFvs were chosen from two molecules that are currently in clinical trials and are well documented and tested for stability and production. The anti-CD19 and anti-CD3 scFv were directly adopted from the BiTE molecule blinatumomab. The antiCD3 scFv was chosen in the VH-VL orientation, consistent with what used in BiTE. The benchmark molecule was an scFv molecule based on BiTE (v891). AlbuCORE_1 (ABH2) CD3/CD19 fusions were created by attaching the antiCD3 warhead to the natural N terminus of fragment 1 and the antiCD19 to the C terminus of fragment 2 (v1092, polypeptide sequences corresponding to SEQ ID NO:264 and 266). The linkers used were identical to the ones used for the multivalent HER2 AlbuCORE experiments: GGGS (SEQ ID NO: 278) at the N terminus of fragment 1 and (GGSG) 4GG (SEQ ID NO: 279) at the C terminus of fragment 2. A second molecule was created where the warheads were reversed (i.e. anti-CD19 warhead at the natural N terminus of fragment 1 and the anti-CD3 at the C terminus of fragment 2, v1093. v1094 was designed to accommodate two different fusions at the natural termini of the albumin polypeptide (polypeptide sequences corresponding to SEQ ID NO:268). The scFv fusions were linked to the albumin molecule through a GGS linker at the N terminus and a GGSG (SEQ ID NO: 280) linker at the C terminus. The length of the linkers reflect the ones used in the MM-111 molecule, despite having a different sequence type.
Example 11: Heteromultimers with an Albumin Scaffold Bind Specifically to CD3- or CD19-Expressing Cells
[0421] The ability of Anti-CD3×CD19 loaded AlbuCORE-1 (v1092) to CD3+ and CD19+ cells was assessed using FACS and compared to WT-HSA loaded with the same anti-CD scFvs (v1094).
[0422] The results are shown in FIG. 4 and demonstrate that both v1092 and v1094 are able to bind to CD3-expressing Jurkat T-cells and to CD19-expressing Raji B-cells.
Example 12: Heteromultimers with Heterodimeric Fc or Albumin Scaffolds Show Comparable B-Cell Targeting and T-Cell Bridging
Example 13: Exemplary Heteromultimers have Higher Anti-CD3 K.SUB.D .and Higher Bmax in Binding to T and B Cell as Determined by FACS
Example 14: Heteromultimer v875 is Able to Bridge Jurkat CD3 T Cells and Raji CD19 B Cells
Example 15: Bridging of B and T Cells by Heteromultimers is Robust at Varying Antibody Concentrations or Cell Ratios
Example 16: Bridging of B and T Cells is Robust Across Differently Engineered Heteromultimers Constructs
Example 17: Effects of v875, v1380, v1379 on IL-2 Activated and Resting CD20+, CD4+, CD8+Subsets
Example 18: Exemplary Heteromultimers v875 and v873 Require the Presence of Both Effector T Cell and Target B Cells to Mediate Cytotoxic Effects
Example 19: Exemplary Heteromultimers can Mediate ADCC or Impaired ADCC to Target Daudi B Cells
[0444] Antibody-dependent cell-mediated cytotoxicity assays (ADCC) were performed with v875, v1379 and v1380 using Daudi cells as target B cells and FcRy3a immobilized NK92 cells as the effector cells (GS193761) by the following method.
Example 20: Exemplary Heteromultimers have Impaired CDC-Mediated Lysis of Daudi B Cells
[0449] Cell based complement dependent cytotoxicity assays (CDC) were performed with v875, v1379 and v1380 using Daudi cells as target B cells. Human serum from healthy donors (NHS) was used as the source of complement. 10 μl NHS (10% final concentration in 40 reaction volume) were added to each well to initiate the CDC cascade and incubated for 2 hours. Cell viability was measured with CellTiter-Glo@ Luminescent Cell Viability Assay Kit.
Example 21: Cell Proliferation and Cytokine Release Assessment of Exemplary Heteromultimers
Example 22: Exemplary Heteromultimers can Bridge Two or More Target B Cells Per Effector T Cell
[0465] The ability and ratio of numbers of T cell bridged to B cells was examined with the exemplary heteromultimer v875 by microscopy using the method described in Example 3 with the following modifications.
[0466] Labeled Raji B cells (red) and labeled Jurkat T cells (blue) were incubated for 30 min at RT with 3 nM of human IgG or v875. The cell suspension was concentrated by removing 180 μl of supernatant. Cell were resuspended in the remaining volume and imaged at 200× and 400×.
[0467] FIG. 30 shows the results from the T:B cell bridging microscopy comparing v875 and human IgG (3 nM) at 200× and 400× magnification; the phase image (top panel), fluorescence image (middle panel) and inverted fluorescence (bottom panel) are presented. FIG. 30A shows a direct comparison of human IgG and v875 at 200× magnification and illustrates a higher amount of bridging visible between Raji B cell and Jurkat T cells compared to human IgG. FIG. 30B and FIG. 30C show two fields of view for v875 (FIG. 30B) and human IgG (FIG. 30C) at 400× magnification. FIG. 30B shows images of v875-mediated immune complex formation between Jurkat T cells (dark grey cells in fluorescence inverted image) and Raji B cells (light grey cells in fluorescence inverted image), and that one Jurkat T cell can bridge 1-3 Raji B cells. FIG. 30C shows images following human IgG incubation with Jurkat T cells and Raji B cells. FIG. 30C shows an absence of the Jurkat-Raji bridging following incubation with human IgG, compared to the v875-mediated Jurkat: Raji bridging that is visible in FIG. 30B.
Example 23: Exemplary Heteromultimer Binding to Fcγ Receptors as Assessed by Surface Plasmon Resonance
Example 25: Expression and Purification of Heteromultimers
[0476] Description of the methods used in the expression and purification of exemplary heteromultimer are described in Example 2.
Example 26: Exemplary Heteromultimers can be Purified to >99% Heterodimer Purity and <1% Aggregates
Example 27: Exemplary Heteromultimers have a CH3 Tm that is Greater than 75° C.
[0487] CH3 domain stability of exemplary heteromultimers was examined by DSC using the following method. All DSC experiments were carried out using a GE VP-Capillary instrument. The proteins were buffer-exchanged into PBS (pH 7.4) and diluted to 0.3 to 0.7 mg/mL with 0.137 mL loaded into the sample cell and measured with a scan rate of 1° C./min from 20 to 100° C. Data was analyzed using the Origin software (GE Healthcare) with the PBS buffer background subtracted.
[0488] The DSC results shown in FIGS. 33A, B and C show that v875 has an estimated CH3 Tm >76° C. (FIG. 33A), v1380 has an estimated CH3 Tm >82.3° C. (FIG. 33B), and v1379 has an estimated CH3 Tm >82.5° C. (FIG. 33C).
Example 28: Design, Expression and Purification of CD3/CD20 and Additional CD3/CD19 Heteromultimer Constructs
[0489] V5850 (corresponding to polypeptide sequences SEQ ID NOs: 203, 205 and 207), v5851 (corresponding to polypeptide sequences SEQ ID NOs: 209, 211 and 213), v5852 (corresponding to polypeptide sequences SEQ ID NOs: 215, 217 and 219), v6324 (corresponding to polypeptide sequences SEQ ID NOs: 221 and 223), v6325 (corresponding to polypeptide sequences SEQ ID NOs: 225, 227 and 229), v1813 (corresponding to polypeptide sequences SEQ ID NOs: 231, 233 and 235), v1821 (corresponding to polypeptide sequences SEQ ID NOs: 237, 239 and 241), v1823 (corresponding to polypeptide sequences SEQ ID NOs: 243, 245 and 247) exemplify bispecific CD3/CD19 or CD3/CD20 hybrid heterodimeric Fc constructs. Bispecific hybrid variants are composed of a F(ab′) on either chain A or B paired with an scFv-Fc on the alternate polypeptide chain. Chain A of the heterodimer Fc is comprised of the following mutations: T350V_L351Y_F405A_Y407V and Chain B of the heterodimer Fc is comprised of the following mutations: T350V_T366L_K392L_T394W. v6324 exemplifies bispecific CD3/CD20 scFv heterodimeric Fc constructs. V1813, v1821, and v1823 exemplify CD3/CD20 common light chain heterodimeric Fc constructs. Common light chain variants are composed of two different F(ab′) s, each on complimentary heterodimer Fc, which share a single light chain. The specific variant composition is indicated in Table 7.
However, the specification does not teach the structure, e.g., amino acid sequence of the heavy and light chain variable region of all antigen-binding domain, e.g., any scFv (claim 3), Fab (claim 4), antibody (claim 8), (humanized or monoclonal antibody (claim 9), multispecific antibody (claim 10) or bispecific antibody (claim 11) that bind to any and all potential different antigen (claim 19) and any CD3 as a pharmaceutical composition (claim 14) to enable one of skilled in the art to make and use without undue experimentation.
It is known in the art that antibodies have a large repertoire of distinct structures and that a huge variety of antibodies can be made to bind to a single epitope.
For example, Lloyd et al (of record, Protein Engineering, Design & Selection 22:159-168, 2009; PTO 1449) teach that hundreds of functional antibody fragments can be isolated from an antibody library that bind to the same antigen wherein these antibodies have distinct heavy and light chain sequences, see, e.g., Discussion.
Similarly, Edwards et al (of record, J Mol Biol 334(1): 103-118, 2003; PTO 1449) find that over 1000 antibodies, all different in amino acid sequence, were generated to a single protein; 568 different amino acid sequences identified for the V(H) CDR3 domains of these antibodies (Abstract).
Poosarla et al (of record, Biotechn. Bioeng., 114(6): 1331-1342, 2017; PTO 892) teach substantial diversity in designed mAbs (sharing less than 75% sequence similarity to all existing natural antibody sequences) that bind to the same 12-mer peptide, binding to different epitopes on the same peptide. Said reference further teaches “most B-cell epitopes... in nature consist of residues from different regions of the sequence and are discontinuous...de novo antibody designs against discontinuous epitopes present additional challenge.
Even assuming the antibody binds to CD3, Shepard et al (of record, PLOSOne 18(6): e0273884, June 22, 2023; PTO 892) teach the unpredictability of swapping CD3 binding domains in the context of a BiTE in so far as the binding requirement to CD3 epsilon much less the extracellular epitope:
“We next sought to test our platform for flexibility with respect to screening novel T-cell engaging domains of the bispecific molecule. We developed a number of novel mouse monoclonal antibodies (mAbs) against human CD3 complex using a multi-antigen immunization strategy in mice and traditional hybridoma screening. Through this process, we were able to identify several monoclonal antibodies with reactivity to Jurkat cells (Fig 4A) and human T cells (Fig 4B). To assess whether scFvs derived from novel CD3-targeted mAbs would be functional within as part of a BiTE molecule, we cloned 4 anti-human CD3 single chain variable fragments into CD19 or EGFRvIII specific BiTE plasmids (Fig 4C). We then generated supernatants using transient transfection of HEK293T as described above. BiTE supernatants were screened for activity using Jurkat cells in co-culture with EGFRvIII-expressing U87-VIII targets or CD19-expressing Raji cells. Previously tested constructs using OKT3 CD3-engager arms showed activity with both EGFRvIII and CD19 specific BiTEs, whereas we detected activity for only one of our novel CD3-engager BiTEs and only when combined with an EGFRvIII-specific scFv (Fig 4D). To confirm these results, we repeated BiTE production and Jurkat co-culture screening of CD19 and EGFRvIII targeted BiTE molecules incorporating OKT3 or the novel 1E2 CD3-targeting single chain variable fragment. Whereas EGFRvIII BiTEs incorporating an scFv derived from the 1E2 mAb or OKT3 showed specific reactivity against EGFRvIII expressing U87-vIII cells, only CD19-OKT3 showed reactivity to CD19-expressing Raji cells (Fig 4E). These results indicate that the novel CD3-targeting 1E2-scFv is functional only for an EGFRvIII-targeting BiTE but not a CD19-targeting BiTE, likely due to the specific binding characteristics of the CD19 or EGFRvIII scFv elements.”
“Similarly, we also find that only one of the novel CD3-targeting scFvs tested here showed activity in BiTE format, and even then, activity was restricted to combination with EGFRvIII-targeting scFv and not with a CD19-specific scFv. Again, we have no insight into the failure rate for BiTE molecules in this assay, as the intent of this assay is to provide a rapid means for testing biological activity rather than focusing on various aspects of antibody characterization. We are currently undertaking molecular studies to test whether incorporating different linker domains may be able to improve the activity of BiTEs using the novel CD3-scFv reported to have activity here, something that may provide further insight into the design constraints for these BiTE molecules.”
Given that hundreds of unique antibody structures may bind a single antigen, the structure of an antibody cannot be predicted from the structure of the antigen, and a single species, or small group of species, cannot define a structure-function relationship so as to be representative of all the antibodies that bind to that antigen.
Regarding pharmaceutical composition (claim 14), the specification does not teach a representative number of species of antibody that correlated with binding to which antigen is effective as a pharmaceutical composition for treating which disease. There are insufficient in vivo working example. It is unpredictable which antibody heterodimer is effective as a pharmaceutical composition for treating which disease.
Second, in response to the argument that amended claim 1 relates to a specific combination of mutations (a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution) that, when incorporated into the CH2 domain of an antibody, silence FcγR binding, the specification discloses a first polypeptide construct and a second polypeptide construct each comprises a human IgG1 Fc wherein the first polypeptide construct (chain A) comprising a L234A, a L235A, a D265S, T350V, L351Y, F405A and Y407V, and wherein the second polypeptide construct (chain B) comprising a L234A, a L235A, a D265S, T350V, T366L, K392L, and T394W, wherein the amino acid positions are numbered according to the EU index of Kabat, see para. [0359] to [0366], [0489] above.
However, the specification does not teach heterodimeric Fc region comprising a first polypeptide construct and a second polypeptide construct each comprises a human IgG1 Fc comprising a variant CH2 domain, each variant CH2 domain comprising just L234A, a L235A, and a D265S as now claimed in claim 1 where the first and second Fc form heterodimer. The specification does not teach these mutations (a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution) form heterodimeric Fc region as now claimed in claim 1. It is the combination of substitutions recited in claim 6 that form a stable Fc heterodimer. It is the combination of substitution in the CH3 domain such as the first polypeptide construct (chain A) comprising a T350V, L351Y, F405A and Y407V, and wherein the second polypeptide construct (chain B) comprising a T350V, T366L, K392L, and T394W (claim 6) that forms stable heterodimer.
For these reasons, the rejection is maintained.
Claim Rejections - 35 USC § 102
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale or otherwise available to the public before the effective filing date of the claimed invention.
Claims 1, 3-4, 9-10, and 14 are rejected under 35 U.S.C. 102 (a)(1) as being anticipated by US Pat No. 8,101,720 (issued Jan 24, 2012; PTO 1449) as evidenced by Hezareh (J Virology 75(24): 12161-12168, 2001; PTO 1449).
Regarding claims 1, 3-4, the ‘720 patent teaches Fc fusions (see col. 3, line 34) or antibody, e.g., bispecific single chain Fv dimers (see col. 39, line 6) comprising two isolated Fc variant of human IgG1 Fc (see Summary of invention, col. 4, Fig 2a, col. 8, line 58-66, in particular) wherein each of the Fc comprising at least one substitutions such as L234A (col. 16, line 22), L235A (col. 16, line 24), and D265S (see col. 16, line 47), wherein the amino acid positions are numbered according to the EU index of Kabat, see col. 12, lines 40-44. The Fc variant comprises an antibody that is a human IgG1 antibody. The term “comprising” is open-ended. It expands the claimed Fc variant to include additional substitutions.
The ‘720 patent teaches that additional modification in the Fc to form heterodimeric, bifunctional or multifunctional molecule, see col. 39, line 1-12, in particular. The ‘720 patent teaches that the IgG is a human IgG1, see col. 8, line 65-67, col. 20, line 55, col. 22, lines 53-56, col. 38, lines 24-56.
Regarding antigen-binding domain, the ‘720 patent teaches that antigen-binding domains include Fab, or scFv, see col. 23, line 26-11, or chimeric, or humanized or fully human antibody, see col. 27-28, in particular.
Regarding antigen, the ‘720 patent teaches that virtually any antigen may be targeted, e.g., CD3, see col. 29, line 48, in particular.
Evidentiary reference Hezareh teaches double substitutions L234A and L235A in the CH2 domain completely abolished FcγRI (CD64) binding (see p. 12163, Results, p. 12164, in particular), reduced antibody dependent cytotoxicity (ADCC) (see p. 12164, left col, FIG 2, in particular) and completely abashed binding to FcγRIIa (CD32a) and FcγRIIIa (CD16a), see p. 12166, left col.
Regarding claim 3, the ‘720 patent teaches that the antibody comprises scFv joined to the Fc variant (see col. 23, line 11, col. 23, line 32).
Regarding claim 4, the ‘270 patent teaches that the reference Fc dimer comprises an antigen binding domain, e.g., Fab (see para. 7, line 10). The ‘720 patent teaches that the reference Fc variant comprises at least one antigen-binding domain such as full-length antibody (see col. 22, lines 50-51) or Fc fusion protein (see col. 22, line 50).
Examples of antibody include antibody fragment such as Fab as per claim 4 (see col. 23, lines 4-5), a humanized antibody (see col. 27, line 56) as per claim 9.
Regarding claim 9, the ‘720 patent teaches that the antibody is a humanized antibody (see col. 22, line 63).
Regarding claim 10, the ‘720 patent teaches multispecific antibodies, see col. 23, line 25.
Regarding claim 11, the ‘720 patent teaches bispecific antibodies, see col. 22, line 61, col. 23, line 25.
Regarding claim 14, the ‘720 patent teaches pharmaceutical composition comprising antibody or Fc fusion and a pharmaceutically acceptable carrier, see col. 54, lines 55-67.
Thus, the reference teachings anticipate the claimed invention.
Applicant's arguments filed January 14, 2026 have been fully considered but they are not persuasive.
Applicant’s position is that a reference must describe each and every element as set forth in the claim. MPEP § 2131. Applicant submits that amended claim 1 is not anticipated by the cited references because they fail to disclose each and every element as set forth in amended claim 1.
The '720 patent lists over 700 individual mutations that may be made in the Fc region (see columns 16-17) and describes certain combinations of mutations that may be included in an Fc variant (see, for example, columns 18-20). None of these combinations include the mutations L234A, L235A or D265S. Nowhere does the '720 patent disclose the specific combination of mutations recited in amended claim 1. Hezareh was relied on only for disclosing the double mutations L234A/L235A and thus fails to cure the deficiency of the '720 patent.
At least for the foregoing reasons, neither of the cited references, individually or in combination, anticipate amended claim 1 nor claims dependent therefrom. Accordingly, reconsideration and withdrawal of the rejection is respectfully requested.
In response to applicant’s argument that there is no teaching in the ‘720 patent for the combination of mutations L234A, L235A and D265S, the term “comprising” is open ended. The mutation in the Fc may include additional substitutions. The ‘720 patent teaches Fc fusions (see col. 3, line 34) or antibody, e.g., bispecific single chain Fv dimers (see col. 39, line 6) comprising two isolated Fc variant of human IgG1 Fc (see Summary of invention, col. 4, Fig 2a, col. 8, line 58-66, in particular) wherein each of the Fc comprising at least one substitutions such as L234A (col. 16, line 22), L235A (col. 16, line 24), and D265S (see col. 16, line 47), wherein the amino acid positions are numbered according to the EU index of Kabat, see col. 12, lines 40-44.
For these reasons, the rejection is maintained.
Claims 1, 3-4, 6, 9, and 14 are rejected under 35 U.S.C. 102 (a)(1) as being anticipated by Escobar-cabrera (US20160102135, claimed priority to 61/829,973 filed May 31, 2013; PTO 892).
Regarding claim 1, Escobar-cabrera teaches isolated dimer, e.g., heterodimer comprising an human IgG1 Fc (para. [0093]) having two Fc polypeptides each comprising a variant CH2 (see para. [0081]) and a CH3 (see entire document, Summary of invention, para. [0052], [0071], [0088]) wherein each Fc comprises a L234A, a L235A (see para. [0025], [0026], [0028], [0029]), D265S (see para. [0030]). The antigen-binding domain binds to a target, e.g., CD3, see para. [0191].
Regarding claims 3-4, Escobar-cabrera teaches that the heterodimer further comprises at least one antigen-binding construct fused to the IgG Fc construct. In certain embodiments, the at least one antigen-binding construct is selected from a Fab fragment, an scFv, see para. [0039], [0074].
Regarding claim 6, Escobar-cabrera teaches that the first Fc chain A comprises T350V, L351Y, F405A, and Y407V, and the second Fc chain B comprises amino acid substitutions T350V, T366L, K392M and T394W, see para. [0234]. The term comprises is open ended. It expands the chain A to include S400E and chain B to include N390R.
Regarding claim 9, Escobar-cabrera teaches that the antibody is a monoclonal (see para. [0111]), a humanized monoclonal antibody (see para. [01231]).
Regarding claims 13 and 14, Escobar-cabrera teaches pharmaceutical composition comprising antibody or Fc fusion and a pharmaceutically acceptable carrier, see para. [0077].
Thus, the reference teachings anticipate the claimed invention.
Applicant's arguments filed January 14, 2026 have been fully considered but they are not persuasive.
Applicant submits that amended claim 1 is not anticipated by the cited references because they fail to disclose each and every element as set forth in amended claim 1.
Escobar-Cabrera discloses heteromultimers comprising two Fc polypeptides, each Fc polypeptide comprising a modified lower hinge region, where the modified hinge region comprises asymmetric amino acid modifications, i.e. the amino acid modifications in the hinge region of one Fc polypeptide are different from those in the hinge region of the other Fc polypeptide (see paragraph [0054] of Escobar-Cabrera). Escobar-Cabrera does not disclose an antibody comprising "a first polypeptide construct and a second polypeptide construct, each polypeptide construct comprising a human IgG Fc region sequence comprising a variant CH2 domain, each variant CH2 domain comprising a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution" as required by amended claim 1. Thus, Escobar-Cabrera fails to teach each and every element of the claimed invention.
In response, the term “comprising” is open ended. It expands the Fc to include additional mutation. Escobar-cabrera teaches heterodimer comprising an human IgG1 Fc (para. [0093]) having two Fc polypeptides each comprising a variant CH2 (see para. [0081]) and a CH3 (see entire document, Summary of invention, para. [0052], [0071], [0088]) wherein each Fc comprises L234A, L235A (see para. [0025], [0026], [0028], [0029]), D265S (see para. [0030]). The antigen-binding domain binds to a target, e.g., CD3, see para. [0191].
Regarding claims 3-4, Escobar-cabrera teaches that the heterodimer further comprises at least one antigen-binding construct fused to the IgG Fc construct. In certain embodiments, the at least one antigen-binding construct is selected from a Fab fragment, an scFv, see para. [0039], [0074].
In response to applicant’s argument that Escobar-Cabrera discloses heteromultimers comprising two Fc polypeptides, each Fc polypeptide comprising a modified lower hinge region, where the modified hinge region comprises asymmetric amino acid modifications, asymmetric amino acid modification, the term “comprising” is open ended. It expands addition to modification in the Fc. Furthermore, it is noted that the title of instant application discloses bispecific asymmetric heterodimers comprising anti-CD3 constructs. If applicant’s substitutions L234A, L235A and D265S can form Fc heterodimer, so is the reference’s. If the reference’s substitutions L234A, L235A and D265S cannot form Fc heterodimer, so is applicant’s.
For these reasons, the rejection is maintained.
New Ground of Rejection Necessitated by Amendment filed January 14, 2026
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 10 and 11 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which applicant regards as the invention.
Claim 10 recites the limitation "the antibody is multispecific" in claim 1. There is insufficient antecedent basis for this limitation in the claim. The antibody in Claim 1 wherein one of the first and second polypeptide construct comprises just a CD3 antigen-binding domain that binds to CD3.
Claim 11 recites the limitation "the antibody is bispecific" in claim 1. There is insufficient antecedent basis for this limitation in the claim.
Conclusion
SEQ ID NO: 133, 134, 137, 138, 141, 142, 38, 40, 42, 44, 46, 48 are free of prior art.
No claim is 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.
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/PHUONG HUYNH/ Primary Examiner, Art Unit 1641
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